PMID- 19120020 TI - Changes in physicochemical parameters and alpha-crystallin expression in the lens during cataract development in OXYS rats. 5. AB - The pathogenesis of cataract is associated with oxidative stress and with altered crystallin expression but it is still understood incompletely. In this study, the senescence-accelerated OXYS rats were used as a model. The first biomicroscopic signs of cataract in OXYS rats were registered at the age of 1.5 months; at 3 months morbidity reached 90%, and at 6 months it reached 100%. Cataract manifestation progresses: at 24 months mature cataract was detected in 90% of eyes of OXYS rats, whereas in 80% of Wistar rat eyes only initial signs of this disease were detected. Analysis of lens redox-parameters has shown that in OXYS rats the intensity of tryptophan fluorescence is higher, the GSH content being higher at 2 months but during formation of mature cataract at 13, 18, and 24 months being lower than in Wistar rats. Decrease in solubility of OXYS rat lens proteins was observed at the age of 13 months. At the age of 3 months gene expression of alphaA-crystallin and alphaB-crystallin was 3-fold and 25% lower, respectively, than in Wistar rats. At the age of 14 months there was a 27-fold decrease in expression of alphaB-crystallin in OXYS rats and it became 21-fold lower than in control. Proteins are synthesized in lens epithelial cells and dystrophic changes in senile cataract result in decrease in structural protein expression. The changes observed in OXYS rats are evidently associated with the dystrophic changes in lens epithelium, which we have described earlier, and are consistent with the model of senile cataract. PMID- 19120019 TI - Role of acidosis, NMDA receptors, and acid-sensitive ion channel 1a (ASIC1a) in neuronal death induced by ischemia. AB - This review collects data on the influence of intracellular and extracellular acidosis on neuronal viability and the effect of acidosis on neuronal damage progressing under brain ischemia/hypoxia. Particular attention is devoted to the involvement of ionotropic glutamic receptors and acid-sensitive ion channel 1a in these processes. PMID- 19120021 TI - Functional significance of a putative sp1 transcription factor binding site in the survivin gene promoter. AB - We sequenced 1500-bp genomic DNA regions upstream from the survivin gene (BIRC5). DNA was isolated from human placenta and tumors of patients with diagnosed squamous cancer of the lung that showed high-level BIRC5 gene expression. We have revealed four new promoter allelic variants differing in single nucleotide substitutions, one variant with two nucleotide substitutions, and a variant with a TAAA tetranucleotide insertion. All promoter variants displayed low activity in cells with functionally active p53 protein and high activity in cell lines characterized by low level or absence of p53 protein function. The activity of the promoters with single nucleotide substitutions was comparable to that of the wild-type promoter, whereas two nucleotide substitutions markedly reduced the activity. We also demonstrated the functional significance of a putative Sp1 transcription factor-binding site at (-63...-54) upstream from the transcription initiation site. Mutation within this sequence led to a sharp decrease of promoter activity. The functional architecture of the survivin promoter is discussed based on results known from the literature and those obtained here. PMID- 19120022 TI - Study of spatial organization of chicken alpha-globin gene domain by 3C technique. AB - This work deals with 3C (Chromosome Conformation Capture) analysis of the chicken alpha-globin gene domain in embryonic erythrocytes and lymphoid cells. Ligation products were quantitatively analyzed by real-time PCR with TaqMan probes. It was found that in lymphoid cells, where alpha-globin gene is not active, the domain has a relatively extended configuration. In embryonic erythrocytes that transcribe alpha(D) and alpha(A) genes, simultaneous interaction of several domain elements was revealed including the major regulatory element, the erythroid-specific DNase I hypersensitive site at a distance of 9 kb upstream from the alpha-globin gene cluster (-9 DHS), promoter of the housekeeping gene CGTHBA, the alpha(D)-globin gene promoter, and the erythroid-specific enhancer located after the alpha-globin gene cluster. We suppose that such interaction is necessary to provide for the active transcription status of the chicken alpha globin gene domains in erythroid cells. PMID- 19120023 TI - Comparative plasma membrane-associated proteomics of immortalized human hepatocytes. AB - This work was initiated with the purpose of purifying and identifying differentially expressed plasma membrane-associated proteins between human liver cancer cell line HepG2 and normal liver cell line L02. The combined strategy of sucrose density gradient centrifugation and subsequent phase partition was applied to obtain high-purity proteins of plasma membrane. Two-dimensional gel electrophoresis revealed the differential protein profile between the two cell lines. A total of 13 plasma membrane-associated proteins containing 10 up regulated proteins and three down-regulated proteins in HepG2 cells were successfully identified by MALDI-Q-TOF mass spectrometry; they participate in multiple biological functions such as adhesion, proliferation, apoptosis, and signal transduction. The identified proteins could provide helpful reference in clinical investigations on potential candidates for diagnosis and therapy of liver cancer. PMID- 19120024 TI - DNA polymerases beta and lambda as potential participants of TLS during genomic DNA replication on the lagging strand. AB - The main strategy used by pro- and eukaryotic cells for replication of damaged DNA is translesion synthesis (TLS). Here, we investigate the TLS process catalyzed by DNA polymerases beta and lambda on DNA substrates using mono- or dinucleotide gaps opposite damage located in the template strand. An analog of a natural apurinic/apyrimidinic site, the 3-hydroxy-2-hydroxymetylthetrahydrofuran residue (THF), was used as damage. DNA was synthesized in the presence of either Mg2+ or Mn2+. DNA polymerases beta and lambda were able to catalyze DNA synthesis across THF only in the presence of Mn2+. Moreover, strand displacement synthesis was not observed. The primer was elongated by only one nucleotide. Another unusual aspect of the synthesis is that dTTP could not serve as a substrate in all cases. dATP was a preferential substrate for synthesis catalyzed by DNA polymerase beta. As for DNA polymerase lambda, dGMP was the only incorporated nucleotide out of four investigated. Modified on heterocyclic base photoreactive analogs of dCTP and dUTP showed substrate specificity for DNA polymerase beta. In contrast, the dCTP analog modified on the exocyclic amino group was a substrate for DNA polymerase lambda. We also observed that human replication protein A inhibited polymerase incorporation by both DNA polymerases beta and lambda on DNA templates containing damage. PMID- 19120025 TI - Cloning and functional identification of two novel BRCA1 splicing variants. AB - BRCA1 is an important tumor suppressor gene associated with inherited breast and ovarian cancers. In this investigation, two novel BRCA1 splicing variants were cloned from breast cancer cell line ZR-75-30. These transcripts, named BRCA1-PI21 Delta2-21 and BRCA1-Delta2-14, lacked most exons of full length BRCA1 gene, but maintained the original reading frame. We also demonstrated the presence of BRCA1 PI21-Delta2-21 in several human cell lines. Expression of both variants fused with green fluorescent protein (GFP) showed that they targeted different subcellular compartments in the transfected cells. Viability of the cells expressing both fusion proteins decreased notably compared with the viability of cells expressing only GFP. Fluorescence activated cell sorting assay confirmed that the overexpression of two splicing variants resulted in cell apoptosis. Taken together, the different subcellular localization and cell effects of two BRCA1 splicing variants imply that they can have different biological functions in breast cancer cells. Elucidating the functions of BRCA1 splicing variants would help to understand the exact roles of the BRCA1 gene in tumor suppression. PMID- 19120026 TI - Efflux of potassium ions from cells and spheroplasts of Saccharomyces cerevisiae yeast treated with silver and copper ions. AB - Silver ions induce the efflux of potassium from cells of the yeast Saccharomyces cerevisiae but have no such effect on spheroplasts. Copper ions and the natural fungicide 2-O-3-hydroxyhexanoyl-beta-D-glucopyranosyl-(1-->4)-(6-O-acetyl-beta-D glucopyranosyl-(1-->16)-2,15,16-trihydroxyhexadecanoic acid) induce the efflux of potassium ions from both cells and spheroplasts of S. cerevisiae. Silver and copper ions inhibit the activity of the plasma membrane H+-ATPase during the treatment of both cells and spheroplasts. It is supposed that the inability of silver ions to stimulate potassium efflux from spheroplasts results from damage to some components of K+ transport systems during preparation of spheroplasts. PMID- 19120027 TI - Polyamine as a signaling molecule for controlling an adaptive mutation. AB - In the absence of exogenous polyamines, the polyamine-deficient Escherichia coli mutant shows not only a characteristic dual-phase growth with abnormal growth, growth arrest, and normal growth after mutation, but also a higher expression of the SOS genes than the polyamine-proficient wild type. The interval of the growth arrest is inversely regulated in a polyamine concentration-dependent manner. These results indicate that the polyamines can act as a signal not only for provoking an adaptive mutation, but also for hastening generation of an adaptive mutation. PMID- 19120028 TI - Complexing of glucose oxidase with anti-glucose oxidase antibodies or the F(ab)'(2)/F(ab)' fragments derived therefrom protects both the enzyme and antibody/antibody fragments against glycation. AB - Incubation of Aspergillus niger glucose oxidase with glucose, fructose, or ribose results in remarkable inactivation of the enzyme. Glucose oxidase incubated with the sugars migrated as a diffuse band of low intensity and silver stained poorly after SDS-PAGE. Purified anti-glucose oxidase antibodies and F(ab)'(2) or F(ab)' derived therefrom were effective in restricting the inactivation of the enzyme induced by the sugars, providing up to 90% protection. The sugars also caused remarkable changes in the electrophoretic behavior of anti-glucose oxidase antibodies and the fragments, but complexing with glucose oxidase restricted the changes both in the enzyme and the antibody/antibody fragments. PMID- 19120029 TI - Antibodies against DNA hydrolyze DNA and RNA. AB - In this work, rabbits were immunized with a high polymer DNA complexed with methylated BSA (mBSA) and by mBSA. It is shown that electrophoretically homogeneous preparations of polyclonal antibodies (Ab) from non-immunized rabbits and animals immunized by mBSA do not exhibit catalytic activity. Ab from the blood of rabbits immunized with the DNA-mBSA complex hydrolyzed poly(C) and different RNAs with efficiency exceeding that towards DNA by approximately 3-4 orders of magnitude. Affinity chromatography of the IgG on DNA cellulose separated the Ab into fractions hydrolyzing both RNA and DNA, and for the first time fractions that hydrolyze only RNA were found. Kinetic parameters that characterize the RNA and DNA hydrolysis by initial Ab preparations and their fractions obtained by separation on an affinity sorbent are compared. PMID- 19120030 TI - Mitochondrial matrix fragmentation as a protection mechanism of yeast Saccharomyces cerevisiae. AB - It was shown that separate fragments of the inner mitochondrial compartment (mitoplasts) can exist under a single non-fragmented outer membrane. Here we asked whether fragmentation of the inner mitochondria could prevent rupturing of the outer membrane and release of pro-apoptotic molecules from the mitochondrial intermembrane space into the cytoplasm during mitochondrial swelling. First, we showed that in Saccharomyces cerevisiae yeast addition of amiodarone causes formation of electrically separate compartments within mitochondrial filaments. Moreover, amiodarone treatment of Deltaysp2 mutant produced a higher proportion of cells with electrically discontinuous mitochondria than in the wild type, which correlated with the survival of cells. We confirmed the existence of separated mitoplasts under a single outer membrane using electron microscopy. Mitochondria with fragmented matrixes were also detected in cells of the stationary phase. Our data suggest that such fragmentation acts as a cellular protective mechanism against stress. PMID- 19120031 TI - Identification of recognition sites for myc/max/mxd network proteins by a whole human chromosome 19 selection strategy. AB - In this study, we have identified 20 human sequences containing Myc network binding sites in a library from the whole human chromosome 19. We demonstrated binding of the Max protein to these sequences both in vitro and in vivo. The majority of the identified sequences contained one or several CACGTG or CATGTG E boxes. Several of these sites were located within introns or in their vicinity and the corresponding genes were found to be up- or down-regulated in differentiating HL-60 cells. Our data show the proof of principle for using this strategy in identification of Max target genes, and this method can also be applied for other transcription factors. PMID- 19120032 TI - Breast cancer diagnosis from proteomic mass spectrometry data: a comparative evaluation. AB - The performance results of a wide range of different classifiers applied to proteomic mass spectra data, in a blind comparative assessment organised by Bart Mertens, are reviewed. The different approaches are summarised, issues of how to evaluate and compare the predictions are described, and the results of the different methods are examined. Although the different methods perform differently, their rank ordering varies according to how one measures performance, so that one cannot draw unequivocal conclusions about which is 'best.' Instead, it is clear that what matters is not the method by itself, but the interaction of method and user - the degree of sophistication of the user with a method. Nevertheless, such competitions do serve the useful role of setting (constantly improving) baselines against which new researchers can pit their wits and methods, as well as providing standards against which new methods should be assessed. PMID- 19120033 TI - The use of hi fidelity simulation to enhance nursing students' therapeutic communication skills. AB - Nursing students entering psychiatric settings for clinical practice need a solid foundation of therapeutic communication skills. This article presents an innovative strategy for nursing students to practice therapeutic communication skills with psychiatric patients by using hi fidelity simulation with Laerdal SimMan. Using the SimMan vocal function enabled nurse educators to develop communication algorithms that allowed students to interact with SimMan as they would with psychiatric patients. The SimMan algorithms can be designed to mimic many scenarios typically found in psychiatric settings. Nursing students can use this technology to take the therapeutic communication skills they have learned in the classroom and practice them in a safe laboratory environment before entering actual psychiatric settings. The ability of students to practice communication skills prior to entering psychiatric settings can promote effective therapeutic communication skills and decrease student anxiety. PMID- 19120034 TI - Nursing graduate supervision of theses and projects at a distance: issues and challenges. AB - Nursing graduate supervision of theses and projects at a distance is a new experience for many faculties. In our global and mobile society, nursing students frequently seek graduate programs that are geographically distant from their home communities. As options for nursing graduate education through distributive learning become increasingly available, the challenges for faculty to supervise graduate students at a distance pose issues and concerns. In this paper, key issues including difficulty deciding between a project and a thesis, difficulty identifying a supervisor, developing the mentoring relationship between the student and the supervisor, and conducting analysis at a distance are discussed. Strategies developed to address these challenges are presented and critiqued. PMID- 19120035 TI - Evaluation of an online graduate nursing curriculum: examining standards of quality. AB - Recent offerings of online courses have outpaced the evaluation of the quality of those offerings, particularly at the program level. The purpose of this project was to evaluate the quality of a set of 16 graduate nursing courses developed for three master's specialty programs. An interdisciplinary group of nursing faculty and instructional technologists collaborated in the development of a quality assessment tool and evaluated 16 online graduate nursing courses. Faculty members for each of the courses were interviewed as part of the process. The collaborative design and development process is discussed, as well as evaluation techniques and the data collection instrument. Examples of best practices and areas for improvement in online courses are presented. The findings will help others as they develop and evaluate online curricula. This project provides a model for interdisciplinary collaboration on the evaluation of online programs that can be reviewed, modified, and implemented at other institutions. Instruments are included as supplementary material. PMID- 19120036 TI - A severe form of Noonan syndrome and autosomal dominant cafe-au-lait spots - evidence for different genetic origins. AB - AIM: The clinical overlap among Noonan syndrome (NS), cardio-facio-cutaneous (CFC), LEOPARD and Costello syndromes as well as Neurofibromatosis type 1 is extensive, which complicates the process of diagnosis. Further genotype-phenotype correlations are required to facilitate future diagnosis of these patients. Therefore, investigations of the genetic cause of a severe phenotype in a patient with NS and the presence of multiple cafe-au-lait spots (CAL) spots in the patient and four members of the family were performed. METHODS: Mutation analyses of candidate genes, PTPN11, NF1, SPRED1 and SPRED2, associated with these syndromes, were conducted using DNA sequencing. RESULTS: A previously identified de novo mutation, PTPN11 F285L and an inherited NF1 R1809C substitution in the index patient were found. However, neither PTPN11 F285L, NF1 R1809C, SPRED1 nor SPRED2 segregated with CAL spots in the family. The results indicate that the familial CAL spots trait in this family is caused by a mutation in another gene, distinct from previous genes associated with CAL spots in these syndromes. CONCLUSION: We suggest that the atypical severe symptoms in the index patient may be caused by an additive effect on the F285L mutation in PTPN11 by another mutation, for example the NF1 R1809C or alternatively, the not yet identified gene mutation associated with CAL spots in this family. PMID- 19120037 TI - Abnormal growth in mitochondrial disease. AB - AIM: To review the height and weight of children with mitochondrial disease attending our supra-regional service. METHODS: This was a retrospective audit of cross-sectional data. Height and weight measurements were available in 24 children and adolescents (median 7.86 years; range 1.76-20.5 years) who all had a confirmed diagnosis of mitochondrial disease. Values were converted to SD scores using UK reference data. RESULTS: Patients with mitochondrial disease were short with an overall SD score of -1.97 (95% confidence intervals -2.72 to -1.23 SD). Patients were also thin with a relatively low BMI SDS of -1.07 (95% confidence intervals -1.69 to - 0.07 SD), which fell with advancing years (r =-0.7; p < 0.000). CONCLUSION: Short stature and a progressive reduction in body mass index (BMI) are features of mitochondrial disease in childhood. The mechanisms underlying the abnormal growth in these patients need to be studied in more detail. PMID- 19120038 TI - Parental mental health, education, age at childbirth and child development from six to 18 months. AB - AIM: To investigate the effect six-month parental mental health has on children's six and 18-month development. Parental covariates of age and education were also analysed. METHODS: Through a national random selection, 21 648 babies were selected. Parental self perceived overall mental health was measured using 36 Item Short Form Health Survey (SF-36) and children's development using the Taiwan Birth Cohort Study (TBCS) instrument which measures gross motor, fine motor, language and social dimensions of children's development. RESULTS: Both multiple linear regression and structural equation modeling showed that when the covariates of parental education and age at childbirth were added, the effect parental mental health has on children's development decreases. Additionally, maternal mental health had a more persistent and pervasive effect than paternal mental health. Father's mental health at six months had a delayed effect, in that its influence was seen only with children's development at 18 months. Of the three factors of parental mental health, education and age at childbirth, parental education had the most pervasive and persistent effect on children's development. CONCLUSION: Although parental mental health has an effect on children's development, parental education and age at childbirth are vital confounding factors, which should be considered in future studies. Clinical health care providers should provide childcare resources and instructions to younger, less educated and parents with mental symptoms. PMID- 19120039 TI - Sudden infant death syndrome (SIDS) in African Americans: polymorphisms in the gene encoding the stress peptide pituitary adenylate cyclase-activating polypeptide (PACAP). AB - AIMS: Mice lacking pituitary adenylate cyclase-activating polypeptide (PACAP) are prone to sudden death in the second post-natal week, having respiratory and metabolic disturbances reminiscent of the human Sudden Infant Death Syndrome (SIDS). Here we test the hypothesis that the human PACAP gene is a site of genetic variance associated with SIDS in a cohort of 92 victims and 92 matched controls. METHODS: Using polymerase chain reaction and sequencing, we examined the PACAP gene in 92 SIDS cases (46 Caucasians and 46 African Americans) and 92 race- and gender-matched controls. RESULTS: We found no significant associations between PACAP and SIDS in Caucasians. However, in the African Americans, a non synonymous single nucleotide polymorphism (i.e. an aspartic acid/glycine coding variant, rs2856966) within exon 2 of PACAP was significantly associated with SIDS (p = 0.004), as were haplotypes containing this polymorphism (p < 0.0001). Glycine was three times more likely at this location in the African-American SIDS victims (17 cases) than African-American controls (5 cases). CONCLUSION: These data are the first to suggest an association between a variant within the coding region of the PACAP gene and SIDS. Based on these findings, further investigations are warranted into the functional importance of PACAP signaling in neonatal survival and the role of PACAP-signaling abnormalities in SIDS. PMID- 19120041 TI - Training neonatal skills with simulators? AB - AIM: To compare two different ways of learning (self-study vs. simulation sessions) the adequate steps to resuscitate a neonate in the 5th year undergraduate medical curriculum. METHODS: One hundred and eighty students attending the 5-week paediatrics rotation were enrolled; 115 were invited to participate in this study, but only 45 students completed it. After a 50-min 'neonatal resuscitation' theoretical interactive class, students were randomly assigned into two groups: the first (n = 21) participated in a 30-min supervised self-study session, while the second (n = 24) attended a 30-min neonatal resuscitation session using the Zoe (Gaumard Inc., Miami, FL, USA) simulator. RESULTS: Tests consisting of 50 multiple-choice questions were taken before the theoretical class (pre-theoretical test), before the self-study or simulation session (pre-test) and after this session (post-test). Pre-test and post-test scores were similar in both groups (p = 0.118 and p = 0.263, respectively). CONCLUSION: Simulation-based training of medical students in management of neonatal resuscitation do not led to significant differences on short-term knowledge comparing with traditional method. PMID- 19120042 TI - Neuronal migration disorders: clinical, neuroradiologic and genetics aspects. AB - Disorders of neuronal migration are a heterogeneous group of disorders of nervous system development. One of the most frequent disorders is lissencephaly, characterized by a paucity of normal gyri and sulci resulting in a 'smooth brain'. There are two pathologic subtypes: classical and cobblestone. Six different genes could be responsible for this entity (LIS1, DCX, TUBA1A, VLDLR, ARX, RELN), although co-delection of YWHAE gene with LIS1 could result in Miller Dieker Syndrome. Heterotopia is defined as a cluster of normal neurons in abnormal locations, and divided into three main groups: periventricular nodular heterotopia, subcortical heterotopia and marginal glioneural heterotopia. Genetically, heterotopia is related to Filamin A (FLNA) or ADP-ribosylation factor guanine exchange factor 2 (ARFGEF2) genes mutations. Polymicrogyria is described as an augmentation of small circonvolutions separated by shallow enlarged sulci; bilateral frontoparietal form is characterized by bilateral, symmetric polymicrogyria in the frontoparietal regions. Bilateral perisylvian polymicrogyria results in a clinical syndrome manifested by mild mental retardation, epilepsy and pseudobulbar palsy. Gene mutations linked to this disorder are SRPX2, PAX6, TBR2, KIAA1279, RAB3GAP1 and COL18A1. Schizencephaly, consisting in a cleft of cerebral hemisphere connecting extra-axial subaracnoid spaces and ventricles, is another important disorder of neuronal migration whose clinical characteristics are extremely variable. EMX2 gene could be implicated in its genesis. Focal cortical dysplasia is characterized by three different types of altered cortical laminations, and represents one of most severe cause of epilepsy in children. TSC1 gene could play a role in its etiology. CONCLUSION: This review reports the main clinical, genetical and neuroradiological aspects of these disorders. It is hoped that accumulating data of the development mechanisms underlying the expanded network formation in the brain will lead to the development of therapeutic options for neuronal migration disorders. PMID- 19120044 TI - Antidepressants and suicidality in younger adults--is bipolar illness the missing link? PMID- 19120045 TI - Sub-threshold manic symptoms in recurrent major depressive disorder are a marker for poor outcome. AB - OBJECTIVE: A small but significant proportion of patients with major depressive disorder (MDD) report mild manic symptoms which are below the diagnostic threshold for a hypomanic episode. METHOD: We tested for an association between sub-threshold manic symptoms and clinical outcome in almost 600 patients with recurrent MDD who also had no known family history of bipolar disorder. RESULTS: 9.6% of this large sample had a life-time history of sub-threshold manic symptoms. These patients were significantly more likely to have a history of poor response to antidepressants (OR 2.84; 95% CI 1.23-6.56; P < 0.02) and more likely to have experienced psychosis (OR 2.07; 95% CI 1.05-4.09; P < 0.04). They had also experienced more depressive episodes on average (P = 0.006) and were more likely to have been admitted to hospital (P < 0.03). CONCLUSION: Sub-threshold manic symptoms in patients with recurrent MDD may be a useful clinical marker for poor response to antidepressants and a more morbid long-term clinical course. PMID- 19120046 TI - Pathological dissociation and neuropsychological functioning in borderline personality disorder. AB - OBJECTIVE: Transient, stress-related severe dissociative symptoms or paranoid ideation is one of the criteria defining the borderline personality disorder (BPD). Examinations of the neuropsychological correlates of BPD reveal various findings. The purpose of this study was to investigate the association between dissociation and neuropsychological functioning in patients with BPD. METHOD: The performance on an extensive neuropsychological battery of patients with BPD with (n=10) and without (n=20) pathological dissociation was compared with that of healthy controls (n=30). RESULTS: Patients with pathological dissociation were found to have reduced functioning on every neuropsychological domain when compared with healthy controls. Patients without pathological dissociation were found to have reduced executive functioning, but no other differences were found. CONCLUSION: Pathological dissociation is a clinical variable that differentiates patients with BPD with regard to cognitive functioning. PMID- 19120047 TI - No association of depression and anxiety with the metabolic syndrome: the Norwegian HUNT study. AB - OBJECTIVE: To examine the associations of depression and anxiety with the metabolic syndrome. METHOD: Cross-sectional study of 9571 participants aged 20-89 years in the Nord-Trondelag Health Study (HUNT 2). We assessed anxiety and depression with the Hospital Anxiety and Depression Scale and the metabolic syndrome with the International Diabetes Federation criteria. RESULTS: Despite generous statistical power and use of both continuous and categorical approaches, we found no association between anxiety or depression and the metabolic syndrome in models adjusted for age, gender, educational level, smoking, physical activity and pulse rate. When adjusted for age and gender only, we found a weak positive association for depression when a continuous measure was used, but not at the case level. The findings were similar across sexes, and robust for exclusion of cardiovascular disease and antidepressants. CONCLUSION: In this largest study to date we found no association of anxiety and depression with the metabolic syndrome. PMID- 19120048 TI - Engagement and retention in specialist services for people with personality disorder. AB - OBJECTIVE: To quantify levels of engagement and retention in specialist services for people with personality disorder (PD). METHOD: Demographic and clinical data were collected on referrals to 10 specialist services for people with PD. Follow up data on retention and drop-out from services were collected over the following 30 months. RESULTS: Seven hundred and thirteen (60.1%) of 1186 people referred to services were taken by them, of whom 164 (23.0%) subsequently dropped out prior to the completion of an episode of care. Men, younger people and those with higher levels of personality disturbance were less likely to complete a package of care. CONCLUSION: Specialist community-based services for adults with PD are able to engage most of those that are referred to them, but further efforts need to be made to find ways to engage younger people and men with PD. PMID- 19120049 TI - Where to focus efforts to improve overall ratings of care and willingness to return: the case of Tuscan emergency departments. AB - OBJECTIVES: Both regression and optimization models were used to identify an efficient combination of aspects of care (e.g., comfort of waiting room) necessary to improve global emergency department (ED) patient satisfaction. The approach, based on patient survey data, tends to favor aspects of care with large regression coefficients and those whose current performance is low, because improvements produce a greater effect on global satisfaction. METHODS: The authors used ED patient satisfaction survey data collected between September and October 2007 from a random sample of 5,277 adult patients who visited 43 EDs in Tuscany, Italy. Ordinal logistic regression models were run to predict overall ratings of care and willingness to return using 20 independent variables (i.e., aspects of care). An optimization model was run to increase these two global items to a maximum of 15%. This model minimizes the total combined percentage increase of the aspects of care. Models using all cases (n = 5,277), cases from local hospitals (n = 4,264), and cases from teaching hospitals (n = 1,013) were run. RESULTS: Four aspects selected by the optimization algorithm were in all models: "satisfaction with waiting time,""comfort of the waiting room,""professionalism of physicians" (technical skills), and "level of collaboration between physicians and nursing staff." Most aspects needed a 15% increase to comply with the percentage increases set for the global satisfaction items. The model found that to increase overall ratings of care by 1, 2, or 8%, hospitals would need to focus only on one aspect: "level of collaboration between physicians and nursing staff." The total number of variables increased to six when the improvement in overall ratings of care was set at 15%. To increase 3 or 5% willingness to return, the optimization algorithm found that 6 or 14 aspects, respectively, are needed. An increase of 6% or more was unfeasible. CONCLUSIONS: This approach is only somewhat efficient, as a cost structure is absent. The optimization model assumes that the cost to increase each aspect by 1% is equivalent. By applying this modeling technique we have demonstrated that, at least, two elements are important to consider when developing efficient improvement strategies to increase global satisfaction: 1) the current level of satisfaction of the aspects of care and 2) the importance ascribed to the aspects of care. A third element, the cost to increase the aspects of care, might also be important. However, the impact of this element on the optimal solution is currently unknown. PMID- 19120050 TI - Video as an effective method to deliver pretest information for rapid human immunodeficiency testing. AB - OBJECTIVES: Video-based delivery of human immunodeficiency virus (HIV) pretest information might assist in streamlining HIV screening and testing efforts in the emergency department (ED). The objectives of this study were to determine if the video "Do you know about rapid HIV testing?" is an acceptable alternative to an in-person information session on rapid HIV pretest information, in regard to comprehension of rapid HIV pretest fundamentals, and to identify patients who might have difficulties in comprehending pretest information. METHODS: This was a noninferiority trial of 574 participants in an ED opt-in rapid HIV screening program who were randomly assigned to receive identical pretest information from either an animated and live-action 9.5-minute video or an in-person information session. Pretest information comprehension was assessed using a questionnaire. The video would be accepted as not inferior to the in-person information session if the 95% confidence interval (CI) of the difference (Delta) in mean scores on the questionnaire between the two information groups was less than a 10% decrease in the in-person information session arm's mean score. Linear regression models were constructed to identify patients with lower mean scores based upon study arm assignment, demographic characteristics, and history of prior HIV testing. RESULTS: The questionnaire mean scores were 20.1 (95% CI = 19.7 to 20.5) for the video arm and 20.8 (95% CI = 20.4 to 21.2) for the in-person information session arm. The difference in mean scores compared to the mean score for the in-person information session met the noninferiority criterion for this investigation (Delta = 0.68; 95% CI = 0.18 to 1.26). In a multivariable linear regression model, Blacks/African Americans, Hispanics, and those with Medicare and Medicaid insurance exhibited slightly lower mean scores, regardless of the pretest information delivery format. There was a strong relationship between fewer years of formal education and lower mean scores on the questionnaire. Age, gender, type of insurance, partner/marital status, and history of prior HIV testing were not predictive of scores on the questionnaire. CONCLUSIONS: In terms of patient comprehension of rapid HIV pretest information fundamentals, the video was an acceptable substitute to pretest information delivered by an HIV test counselor. Both the video and the in-person information session were less effective in providing pretest information for patients with fewer years of formal education. PMID- 19120051 TI - Alpha 2A-adrenergic receptor signaling underlies synergistic enhancement of ethanol-induced behavioral impairment by clonidine. AB - BACKGROUND: We tested the hypothesis that central alpha(2A)-adrenergic receptor (alpha(2A)AR) signaling plays a key role in clonidine-ethanol evoked synergistic behavioral impairment. METHODS: Male Sprague-Dawley rats, with intracisternal and jugular vein cannulae implanted 6 days earlier, were tested for drug-induced behavioral impairment. The latter was assessed as the duration of loss of righting reflex (LORR) and rotorod performance every 15 minutes until the rat recovered to the baseline walk criterion (180 seconds). In a separate cohort, c Fos expression in locus coeruleus (LC) and cerebellum was determined as a marker of neuronal activity following drug treatment. RESULTS: Rats that received clonidine (60 microg/kg, i.v.) followed by ethanol (1 g/kg, i.v.) exhibited synergistic impairment of rotorod performance and LORR. The mixed alpha(2A)AR and I(1)-imidazoline receptor agonist clonidine (30, 60, and 90 microg/kg) synergistically and dose-dependently enhanced behavioral impairment elicited by ethanol (1 g/kg). Possible involvement of I(1)-imidazoline receptors was ruled out because selective I(1)-agonist rilmenidine (300 microg/kg, i.v.) did not cause behavioral impairment alone or enhance ethanol-evoked behavioral impairment. Pharmacological blockade of central alpha(2A)AR (RX821002, 0.3 mg i.c.) abolished the synergy between clonidine and ethanol; the behavioral response caused by the drug combination was similar to that caused by ethanol alone. Conversely, involvement of central alpha(2B)AR in the interaction was ruled out because blockade of central alpha(2B)AR (ARC-239) independently evoked a strong sedative effect. Clonidine (60 microg/kg) or ethanol (1 g/kg) alone increased, but their combination decreased, c-Fos levels in LC, while inconsistent c-Fos responses were observed in cerebellum. CONCLUSIONS: Central alpha(2A)AR, but not I(1)-imidazoline or alpha(2B)AR, signaling is implicated in the synergistic enhancement of ethanol-evoked behavioral impairment by clonidine. Although the mechanism of c-Fos response remains to be investigated, this neurochemical response highlights the LC as a neuroanatomical target for clonidine-ethanol behavioral interaction. PMID- 19120052 TI - Changes in the episodic memory and executive functions of abstinent and relapsed alcoholics over a 6-month period. AB - BACKGROUND: It is still unclear whether episodic memory and executive functions capacities can return to normal in abstinent patients over a 6-month period. Furthermore, the role of interim drinking in cognitive recovery is still not well known. Finally, further research is required to specify the predictive value of cognitive abilities at initial testing in the treatment outcome (abstinence or relapse). The aims of the present study were therefore to measure changes in episodic memory and executive functions over a 6-month period in abstinent and relapsed alcoholics and to ascertain whether neuropsychological results at treatment entry can predict treatment outcome at follow-up. METHODS: Fifty-four alcoholic patients and 54 matched control subjects performed baseline neuropsychological tasks assessing episodic memory, executive functions, the slave systems of working memory and attentional abilities. At the follow-up session (i.e., 6 months later), episodic memory and 3 executive functions (inhibition, flexibility, and updating) were re-examined in the alcoholic patients. RESULTS: Results showed that over the 6-month interval, the abstainers' episodic memory and executive performances had returned to normal, whereas the relapsers performed lower than before in the flexibility task. Episodic memory and executive functions recovery was correlated, in abstainers, with drinking history and age respectively. Finally, there was no significant difference regarding neuropsychological scores at baseline between abstainers and relapsers. DISCUSSION: Over the 6-month interval, abstainers normalized episodic memory and executive performances whereas relapsers obtained executive results which were more severely impaired, emphasizing the influence of interim drinking on cognitive changes. Episodic memory, executive functions, the slave systems of working memory and attentional abilities did not appear to be reliable predictors of treatment outcome over a 6-month interval. PMID- 19120054 TI - Sensitivity and tolerance to the hypnotic and ataxic effects of ethanol in adolescent and adult C57BL/6J and DBA/2J mice. AB - BACKGROUND: There is considerable research examining differences in adolescent and adult sensitivity and tolerance to ethanol related behavioral phenotypes. However, the available published data has almost exclusively assessed these behaviors in outbred rats. The present study was conducted using the alcohol preferring inbred mouse strain C57BL/6J (B6) and the alcohol nonpreferring inbred mouse strain DBA/2J (D2) to determine if differences in the sedative and ataxic effects of ethanol exist between adolescents and adults, and to determine whether there are any genetic influences involved therein. METHODS: Adolescent and adult mice of each sex and genotype were given intraperitoneal (i.p.) injections of ethanol (1.5, 1.75, or 4.0 g/kg) or saline and assessed for the loss of righting reflex (LORR) or hind footslips on the balance beam apparatus. These animals were then tested for the development of tolerance to these behaviors on subsequent days. RESULTS: Despite evident pharmacokinetic differences, D2 adolescents were found to be relatively less sensitive to ethanol's hypnotic actions than their adult D2 counterparts. Adolescent and adult B6 animals did not differ. Furthermore, although adult animals appeared to develop significantly greater degrees of tolerance to ethanol-induced hypnosis compared with adolescents, these effects were likely in part related to differences in ethanol absorption/metabolism across time. Taking into account pharmacokinetic differences and the overall poor performance of male adults, adolescent animals were found to be equally if not more sensitive to the motor incoordinating (ataxic) effects of ethanol. Overall, tolerance to these effects varied by age and genotype but appeared to be related to changes in ethanol pharmacokinetics rather than strict behavioral sensitivity. CONCLUSION: The current work suggests that adolescent B6 and D2 inbred mice exhibit ontogenetic differences in sensitivity to ethanol's hypnotic and ataxic effects. Importantly, in some cases age differences emerge as a function of differential ethanol pharmacokinetics. These results extend the current literature examining this critical developmental period in mice and illustrate the benefits of comparing ethanol related developmental differences in different genetic mouse populations. PMID- 19120053 TI - Global-local interference is related to callosal compromise in alcoholism: a behavior-DTI association study. AB - BACKGROUND: Visuospatial ability is a multifactorial process commonly impaired in chronic alcoholism. Identification of which features of visuospatial processing are affected and which are spared in alcoholism, however, has not been clearly determined. We used a global-local paradigm to assess component processes of visuospatial ability and MR diffusion tensor imaging (DTI) to examine whether alcoholism-related microstructural degradation of the corpus callosum contributes to disruption of selective lateralized visuospatial and attention processes. METHODS: A hierarchical letter paradigm was devised, where large global letters were composed of small local letters. The task required identification of target letters among distractors presented at global, local, both, or neither level. Attention was either selectively directed to global or local levels or divided between levels. Participants were 18 detoxified chronic alcoholics and 22 age matched healthy controls. DTI provided quantitative assessment of the integrity of corpus callosal white matter microstructure. RESULTS: Alcoholics generally had longer reaction times than controls but obtained similar accuracy scores. Both groups processed local targets faster than global targets and showed interference from targets at the unattended level. Alcoholics exhibited moderate compromise in selectively attending to the global level when the global stimuli were composed of local targets. Such local interference was less with longer abstinence. Callosal microstructural integrity compromise predicted degree of interference from stimulus incongruency in the alcoholic group. This relationship was not observed for lateral or third ventricular volumes, which are measures of nonspecific cortical volume deficits. CONCLUSION: Global-local feature perception was generally spared in abstinent chronic alcoholics, but impairments were observed when directing attention to global features and when global and local information interfered at stimulus or response levels. Furthermore, the interference-callosal integrity relationship in alcoholics indicates that compromised visuospatial functions include those requiring bilateral integration of information. PMID- 19120055 TI - Sensitization, duration, and pharmacological blockade of anxiety-like behavior following repeated ethanol withdrawal in adolescent and adult rats. AB - BACKGROUND: Repeated ethanol withdrawal sensitizes anxiety-like behavior in adult rats and causes anxiety-like behavior and decreased seizure thresholds in adolescent rats. Current experiments determined if adolescent rats exhibit sensitized anxiety-like behavior, the duration of this effect, if drug pretreatments blocked these effects, and if these effects differed from those seen in adults. METHODS: Male adolescent rats received three 5-day cycles of 2.5% ethanol diet (ED) separated by two 2-day withdrawal periods, continuous 15 days of 2.5%ED, or a single 5-day cycle of 2.5%ED. Male adult rats received three 5 day cycles of either 2.5% or 3.5%ED. These groups were tested 5 hours into the final withdrawal for social interaction (SI) deficits (an index of anxiety-like behavior). Ethanol intake was monitored throughout and blood concentrations were obtained from separate groups of rats. Additionally, adolescent rats were tested for SI 1, 2, 7, 14, and 18 days and adults 1 and 2 days after the final withdrawal. Some adolescent rats were also pretreated with the CRF(1) antagonist CP-154,526, the 5-HT(1A) agonist buspirone, or the benzodiazepine receptor antagonist flumazenil during the first 2 withdrawals. RESULTS: SI was reduced in adolescent rats following repeated withdrawals of 2.5%ED while neither a continuous or single cycle ED exposure caused this effect. Adult rats also had reduced SI following repeated withdrawals from both 2.5% and 3.5%ED. This effect was present up to 1 week following the final withdrawal in adolescents but returned to baseline by 1 day in adults. CP-154,526, buspirone, or flumazenil prevented this reduction in SI in adolescent rats. CONCLUSIONS: Adolescent rats exhibit sensitized anxiety-like behavior following repeated withdrawals at ED concentrations similar to those used in adults. However, this effect is longer lasting in adolescent rats. Drugs modulating CRF, 5-HT, or GABA systems during initial withdrawals prevent the development of anxiety-like behavior otherwise manifest during a final withdrawal in adolescent rats. PMID- 19120056 TI - The language of intoxication: preliminary investigations. AB - BACKGROUND: The extensive vocabulary individuals use to describe alcohol's subjective effects has largely gone unexamined in contemporary alcohol research. The present study examined the language drinkers use to describe their own intoxication. It is argued that this language can provide a more complete characterization of alcohol's subjective effects than is available from existing objective and subjective measures of alcohol use and can inform future self report research. METHOD: Toward this goal, a preliminary, cross-sectional, web based study of the familiarity and usage of current intoxication-related words was conducted in 2 different samples (n = 290 and 146, respectively) of university undergraduates. RESULTS: Exploratory factor analyses using data from the first sample and confirmatory factor analyses using data from the second sample similarly showed that commonly used terms loaded onto 2 factors, which directly reflected the number of drinks required to be considered moderately or heavily intoxicated, respectively. Gender differences were also found in the familiarity and self-use of some terms across both samples. CONCLUSIONS: The findings suggest that alcohol researchers include multiple intoxication-related terms in future self-report research, and to periodically assess current intoxication-related vocabulary considering demographic, generational, and socio cultural differences. PMID- 19120057 TI - Combined scopolamine and ethanol treatment results in a locomotor stimulant response suggestive of synergism that is not blocked by dopamine receptor antagonists. AB - BACKGROUND: Muscarinic acetylcholine receptors (mAChRs) are well positioned to mediate ethanol's stimulant effects. To investigate this possibility, we examined the effects of scopolamine, a receptor subtype nonselective mAChR antagonist, on ethanol-induced stimulation in genotypes highly sensitive to this effect of ethanol. We also investigated whether the dopamine D1-like receptor antagonist, SCH-23390 or the dopamine D2-like receptor antagonist, haloperidol, could block the extreme stimulant response found following co-administration of scopolamine and ethanol. METHODS: Scopolamine (0, 0.0625, 0.125, 0.25, or 0.5 mg/kg) was given 10 minutes prior to saline or ethanol (0.75 to 2 g/kg) to female FAST (Experiment I) or DBA/2J (Experiment II) mice that were then tested for locomotion for 30 minutes. In Experiments III and IV, respectively, SCH-23390 (0, 0.015, or 0.03 mg/kg) was given 10 minutes prior, and haloperidol (0, 0.08, or 0.16 mg/kg) was given 2 minutes prior, to scopolamine (0 or 0.5 mg/kg), followed 10 minutes later by saline or ethanol (1.5 g/kg) and female DBA/2J mice were tested for locomotion for 30 minutes. RESULTS: FAST and DBA/2J mice displayed a robust enhancement of the locomotor effects of ethanol following pretreatment with scopolamine that was suggestive of synergism. SCH-23390 had no effect on the response to the scopolamine + ethanol drug combination, nor did it attenuate ethanol- or scopolamine-induced locomotor activity. Haloperidol, while attenuating the effects of ethanol, was not able to block the effects of scopolamine or the robust response to the scopolamine-ethanol drug combination. CONCLUSIONS: These results suggest that while muscarinic receptor antagonism robustly enhances acute locomotor stimulation to ethanol, dopamine receptors are not involved in the super-additive interaction of scopolamine and ethanol treatment. They also suggest that in addition to cautions regarding the use of alcohol when scopolamine is clinically prescribed due to enhanced sedative effects, enhanced stimulation may also be a concern. PMID- 19120058 TI - Effects of MAOA-genotype, alcohol consumption, and aging on violent behavior. AB - BACKGROUND: Environmental factors appear to interact with a functional polymorphism (MAOA-LPR) in the promoter region of the monoamine oxidase A gene (MAOA) in determining some forms of antisocial behavior. However, how MAOA-LPR modulates the effects of other factors such as alcohol consumption related to antisocial behavior is not completely understood. METHODS: This study examines the conjunct effect of MAOA-LPR, alcohol consumption, and aging on the risk for violent behavior. Recidivism in severe impulsive violent behavior was assessed after 7 to 15 years in a sample of 174 Finnish alcoholic offenders, the majority of whom exhibited antisocial or borderline personality disorder or both, and featured impulsive temperament traits. RESULTS: The risk for committing new acts of violence increased by 2.3% for each kilogram of increase in yearly mean alcohol consumption (p = 0.004) and decreased by 7.3% for every year among offenders carrying the high activity MAOA genotype. In contrast, alcohol consumption and aging failed to affect violent behavior in the low activity MAOA genotyped offenders. MAOA-LPR showed no main effect on the risk for recidivistic violence. CONCLUSIONS: Violent offenders carrying the high activity MAOA genotype differ in several ways from carriers with the low activity MAOA risk allele previously associated with antisocial behavior. Finnish high activity MAOA genotyped risk alcoholics exhibiting antisocial behavior, high alcohol consumption, and abnormal alcohol-related impulsive and uncontrolled violence might represent an etiologically distinct alcohol dependence subtype. PMID- 19120059 TI - Exogenous IL-15 in combination with IL-15R alpha rescues natural killer cells from apoptosis induced by chronic alcohol consumption. AB - BACKGROUND: Chronic alcohol consumption reduces the percentage and number of peripheral natural killer (NK) cells in mice and in humans. The underlying mechanism for these changes is only partly known. We recently found that chronic alcohol consumption inhibits NK cell release from the bone marrow (BM) and that this is associated with a decrease in splenic NK cells. The number of peripheral NK cells is tightly controlled by homeostatic proliferation. It is not known whether this mechanism is initiated in response to the reduction in splenic NK cells, or if so, why the steady state levels of NK cells are not restored. METHODS: To examine this mechanism, female C57BL/6 mice were given 20% w/v alcohol in the drinking water for 3 months. NK cell proliferation and apoptosis were determined before and after treatment with IL-15 alone or combined with its alpha receptor. RESULTS: Chronic alcohol consumption invoked homeostatic proliferation of splenic NK cells in an attempt to return NK cells to normal levels; however, this did not happen due to enhanced apoptosis of NK cells relative to proliferation. Chronic alcohol consumption decreased IL-15 producing cells in the spleen but not in the BM. The numbers of NK cells in the alcohol consuming mice returned to normal levels in the spleen and were higher than normal in the BM after 2 daily injections of IL-15; however, the enhanced rate of apoptosis due to alcohol consumption was not decreased in the spleen or BM. Combined IL-15 and IL-15R alpha treatment decreased apoptosis of NK cells from alcohol-consuming mice to levels similar to untreated water-drinking mice and greatly increased the percentage and number of NK cells in both the spleen and BM. CONCLUSION: Chronic alcohol consumption causes a self-unrecoverable loss of NK cells in the spleen by compromising NK cell release from the BM and enhancing splenic NK cell apoptosis that can be reversed with IL-15/IL-15R alpha treatment. PMID- 19120060 TI - The overlap in predicting alcohol outcome for two measures of the level of response to alcohol. AB - BACKGROUND: Two different measures have been used to establish a person's level of response (LR) to alcohol as a risk factor for alcohol use disorders. LR values established by the alcohol challenge protocol and the Self-Report of the Effects of Ethanol (SRE) questionnaire usually correlate at 0.3 to 0.4, up to 0.6. However, it is not clear how this correlation relates to the ability of each measure to predict alcohol outcomes. This paper evaluates that overlap. METHODS: Sixty-six Caucasian males (mean age = 22 years) from 2 protocols participated in alcohol challenges with 0.75 ml/kg of ethanol, filled out the SRE, and were followed with a structured interview approximately 5 years later. The relationship between the subjective feelings of intoxication at the time of peak breath alcohol levels from the alcohol challenge and the SRE score for a time early in the drinking career were evaluated regarding predicting the drinks per occasion in the 6 months prior to follow-up. RESULTS: Cross-sectional correlations between alcohol challenge and SRE LR's ranged from -0.25 (p < 0.05) to -0.32 (p = 0.02) for the full sample, and the 2 LR measures correlated with drinking at follow-up (-0.26 and 0.41, respectively). The SRE measure was more robust than the challenge in a regression analysis predicting the outcome in the context of other baseline predictors (e.g., drinking at baseline). As much as 60% of the ability of the more well established (gold standard) alcohol challenge LR to predict outcome was shared with the SRE. The alcohol challenge accounted for as much as 44% of the ability of the SRE to predict outcome. CONCLUSIONS: The SRE generated LR overlapped considerably with the alcohol challenge LR in the ability to predict future heavier drinking. PMID- 19120061 TI - Phosphatidylcholine reverses ethanol-induced increase in transepithelial endotoxin permeability and abolishes transepithelial leukocyte activation. AB - BACKGROUND: Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver disease (ALD). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol enhanced transepithelial permeability of endotoxin and the subsequent transepithelial activation of human leukocytes. METHODS: For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without the addition of CPBS (1.5 mM), PC (0.38 mM), pooled human bile (2%) in combination with ethanol (0 to 66 mM). RESULTS: Ethanol decreased integrity of intestinal epithelial cell monolayer and enhanced transepithelial permeability of endotoxin. Both the transepithelial permeability of endotoxin and the transepithelial stimulation of leukocytes were nearly completely abolished after the apical supplementation of PC with CPBS, but not by CPBS alone. Ethanol up to 66 mM was not able to reverse this effect. CONCLUSIONS: A considerable part of the therapeutic and preventive effect of PC supplementation in ALD might result from a reduction of ethanol-enhanced permeability of endotoxin through the intestinal barrier. PMID- 19120062 TI - Alcohol and colorectal cancer: the role of alcohol dehydrogenase 1C polymorphism. AB - BACKGROUND: Chronic alcohol consumption is a risk factor for colorectal cancer. Animal experiments as well as genetic linkage studies in Japanese individuals with inactive acetaldehyde dehydrogenase leading to elevated acetaldehyde concentrations following ethanol ingestion support the hypothesis that acetaldehyde may be responsible for this carcinogenic effect of alcohol. In Caucasians, a polymorphism of alcohol dehydrogenase 1C (ADH1C) exists resulting in different acetaldehyde concentrations following ethanol oxidation. METHODS: To evaluate whether the association between alcohol consumption and colorectal tumor development is modified by ADH1C polymorphism, we recruited 173 individuals with colorectal tumors diagnosed by colonoscopy and 788 control individuals without colorectal tumors. Genotyping was performed using genomic DNA extracted from whole blood followed by polymerase chain reaction. RESULTS: Genotype ADH1C*1/1 was more frequent in patients with alcohol-associated colorectal neoplasia compared to patients without cancers in the multivariate model controlling for age, gender, and alcohol intake (odds ratio = 1.674, 95% confidence interval = 1.110-2.524, 2-sided p from Wald test = 0.0139). In addition, the joint test of the genetic effect and interaction between ADH1C genotype and alcohol intake (2 sided p = 0.0007) indicated that the difference in ADH1C*1 polymorphisms between controls and colorectal neoplasia is strongly influenced by the alcohol consumption and that only individuals drinking more than 30 g ethanol per day with the genotype ADH1C*1/1 had an increased risk for colorectal tumors. CONCLUSIONS: These data identify ADH1C homozygosity as a genetic risk marker for colorectal tumors in individuals consuming more than 30 g alcohol per day and emphasize the role of acetaldehyde as a carcinogenic agent in alcohol-related colorectal carcinogenesis. PMID- 19120063 TI - Long-term ethanol exposure impairs neuronal differentiation of human neuroblastoma cells involving neurotrophin-mediated intracellular signaling and in particular protein kinase C. AB - BACKGROUND: Revealing the molecular changes in chronic ethanol-impaired neuronal differentiation may be of great importance for understanding ethanol-related pathology in embryonic development but also in the adult brain. In this study, both acute and long-term effects of ethanol on neuronal differentiation of human neuroblastoma cells were investigated. We focused on several aspects of brain derived neurotrophic factor (BDNF) signaling because BDNF activates the extracellular signal-regulated kinase (ERK) cascade, promoting neuronal differentiation including neurite outgrowth. METHODS: The effects of ethanol exposure on morphological differentiation, cellular density, neuronal marker proteins, basal ERK activity, and ERK responsiveness to BDNF were measured over 2 to 4 weeks. qRT-PCR and Western blotting were performed to investigate the expression of neurotrophin receptor tyrosin kinase B (TrkB), members of the ERK cascade, protein kinase C (PKC) isoforms and Raf-Kinase-Inhibitor-Protein (RKIP). RESULTS: Chronic ethanol interfered with the development of a neuronal network consisting of cell clusters and neuritic bundles. Furthermore, neuronal and synaptic markers were reduced, indicating impaired neuronal differentiation. BDNF mediated activation of the ERK cascade was found to be continuously impaired by ethanol. This could not be explained by expressional changes monitored for TrkB, Raf-1, MEK, and ERK. However, BDNF also activates PKC signaling which involves RKIP, which finally leads to ERK activation as well. Therefore, we hypothesized that ethanol impairs this branch of BDNF signaling. Indeed, both PKC and RKIP were significantly down-regulated. CONCLUSIONS: Chronic ethanol exposure impaired neuronal differentiation of neuroblastoma cells and BDNF signaling, particularly the PKC-dependent branch. RKIP, acting as a signaling switch at the merge of the PKC cascade and the Raf/MEK/ERK cascade, was associated with neuronal differentiation and significantly reduced in ethanol treatment. Moreover, PKC expression itself was even more strongly reduced. In contrast, members of the Raf 1/MEK/ERK cascade were less affected and the observed changes were not associated with impaired differentiation. Thus, reduced RKIP and PKC levels and subsequently reduced positive feedback on ERK activation provide an explanation for the striking effects of long-term ethanol exposure on BDNF signal transduction and neuronal differentiation, respectively. PMID- 19120064 TI - Genomewide SNP screen to detect quantitative trait loci for alcohol preference in the high alcohol preferring and low alcohol preferring mice. AB - BACKGROUND: The high and low alcohol preferring (HAP1 and LAP1) mouse lines were selectively bred for differences in alcohol intake. The HAP1 and LAP1 mice are essentially noninbred lines that originated from the outbred colony of HS/Ibg mice, a heterogeneous stock developed from intercrossing 8 inbred strains of mice. METHODS: A total of 867 informative SNPs were genotyped in 989 HAP1 x LAP1 F2, 68 F1s, 14 parents (6 LAP1, 8 HAP1), as well as the 8 inbred strains of mice crossed to generate the HS/Ibg colony. Multipoint genome wide analyses were performed to simultaneously detect linked QTLs and also fine map these regions using the ancestral haplotypes. RESULTS: QTL analysis detected significant evidence of association on 4 chromosomes: 1, 3, 5, and 9. The region on chromosome 9 was previously found linked in a subset of these F2 animals using a whole genome microsatellite screen. CONCLUSIONS: We have detected strong evidence of association to multiple chromosomal regions in the mouse. Several of these regions include candidate genes previously associated with alcohol dependence in humans or other animal models. PMID- 19120065 TI - Ethanol-induced conditioned taste avoidance: reward or aversion? AB - BACKGROUND: Rats avoid intake of a palatable taste cue when paired with all drugs of abuse tested. Evidence suggests that, at least for morphine and cocaine, rats avoid the taste cue because they are anticipating the rewarding properties of the drug. Thus, the suppressive effects of a rewarding sucrose solution and cocaine, but not those of the putatively aversive agent, lithium chloride (LiCl), are exaggerated in drug-sensitive Lewis rats. Likewise, the suppressive effects of sucrose and morphine, but not those of LiCl, are eliminated by bilateral lesions of the gustatory thalamus. Unlike morphine and cocaine, it is less clear whether rewarding or aversive drug properties are responsible for ethanol-induced suppression of intake of a taste cue. The present set of studies tests whether, like cocaine, ethanol-induced suppression of intake of a taste cue also is greater in the drug-sensitive Lewis rats and whether the suppressive effects of the drug are prevented by bilateral lesions of the taste thalamus. METHODS: In Experiment 1, fluid-deprived Lewis and Fischer rats were given 5-minute access to 0.15% saccharin and then injected with saline or a range of doses of ethanol (0.5, 0.75, 1.0, or 1.5 g/kg). There was a total of 6 such pairings. In Experiments 2 and 3, Sprague-Dawley rats received bilateral electrophysiologically guided lesions of the gustatory thalamus. After recovery, suppression of intake of the saccharin cue was evaluated following repeated daily pairings with either a high (1.5 g/kg) or a low (0.75 g/kg) dose of ethanol. RESULTS: Ethanol-induced suppression of intake of the saccharin conditioned stimulus (CS) did not differ between the drug-sensitive Lewis rats relative to the less-sensitive Fischer rats. Lesions of the taste thalamus, however, prevented the suppressive effect of the 0.75 g/kg dose of the drug, but had no impact on the suppressive effect of the 1.5 g/kg dose of ethanol. CONCLUSION: The results suggest that the suppressive effects of ethanol on CS intake are mediated by both rewarding and aversive consequences, varying as a function of dose. PMID- 19120066 TI - Characterization of white matter microstructure in fetal alcohol spectrum disorders. AB - BACKGROUND: Exposure to alcohol during gestation is associated with CNS alterations, cognitive deficits, and behavior problems. This study investigated microstructural aspects of putative white matter abnormalities following prenatal alcohol exposure. METHODS: Diffusion tensor imaging was used to assess white matter microstructure in 27 youth (age range: 8 to 18 years) with (n = 15) and without (n = 12) histories of heavy prenatal alcohol exposure. Voxelwise analyses, corrected for multiple comparisons, compared fractional anisotropy (FA) and mean diffusivity (MD) between groups, throughout the cerebrum. RESULTS: Prenatal alcohol exposure was associated with low FA in multiple cerebral areas, including the body of the corpus callosum and white matter innervating bilateral medial frontal and occipital lobes. Fewer between-group differences in MD were observed. CONCLUSIONS: These data provide an account of cerebral white matter microstructural integrity in fetal alcohol spectrum disorders and support extant literature showing that white matter is a target of alcohol teratogenesis. The white matter anomalies characterized in this study may relate to the neurobehavioral sequelae associated with gestational alcohol exposure, especially in areas of executive dysfunction and visual processing deficits. PMID- 19120068 TI - Alcohol functionally upregulates Toll-like receptor 2 in airway epithelial cells. AB - BACKGROUND: Alcoholics are known to have more severe airway diseases of the lung, such as bronchitis. Little is known about why this phenomenon is observed. We hypothesized that alcohol may modulate Toll-like receptor 2 (TLR2), which regulates inflammation caused by gram-positive bacteria. METHODS: Airway epithelial cells [primary bronchial epithelial cells (NHBE) and 16HBE 14o-] were exposed to 0 to 100 mM alcohol for 0 to 24 hours. Real time PCR was used to quantify TLR2 mRNA. Protein levels of TLR2 were determined using Western blots and fluorescence activated cell sorting (FACS) on cells exposed to 0, 50, and 100 mM alcohol. Finally, cells were "primed" with alcohol, stimulated with a TLR2 agonist (peptidoglycan), and interleukin 8 (IL-8) release was measured. RESULTS: Alcohol, at biologically relevant concentrations (25 to 100 mM), caused a 2 to 3 fold time- and concentration-dependent increase in TLR2 mRNA in normal human bronchial epithelial cells and 16HBE 14o- cells. Western blots for TLR2 revealed a qualitative increase in TLR2 protein in cells exposed to 100 mM alcohol. FACS showed that TLR2 was quantitatively increased on the surface of airway epithelial cells that were exposed to alcohol. Airway cells that were primed with alcohol produced nearly twice as much IL-8 in response to 40 ng of peptidoglycan than naive cells. CONCLUSIONS: Alcohol upregulates TLR2 message and protein in the airway epithelium. This leads to exaggerated inflammation in response to environmental stimuli that would normally be well tolerated in airway epithelial cells. This may be a partial explanation of why alcoholics have more severe airway disease than nonalcoholics. PMID- 19120067 TI - Alcohol primes the airway for increased interleukin-13 signaling. AB - BACKGROUND: Using an experimental model of airway fibrosis following lung transplantation, we recently showed that chronic alcohol ingestion by donor rats amplifies airway fibrosis in the recipient. Associated with alcohol-mediated amplification of airway fibrosis is increased transforming growth factor beta 1(TGFbeta(1)) and alpha-smooth muscle actin expression. Other studies have shown that interleukin-13 (IL-13) modulates TGFbeta(1) signaling during experimentally induced airway fibrosis. Therefore, we hypothesized that IL-13 is a component of alcohol-mediated amplification of pro-fibrotic mediators in the alcoholic lung. METHODS: To test this hypothesis, we analyzed tracheal epithelial cells and type II alveolar cells from control- or alcohol-fed rats, alcohol-treated mouse lung fibroblasts, and human bronchial epithelial cells in vitro for expression of various components of the IL-13 signaling pathway. Signaling via the IL-13 pathway was assessed by measuring levels of phosphorylated signal transducers and activators of transcription-6 (STAT6). In addition, we performed heterotopic tracheal transplantation using control-fed and alcohol-fed donor rats and analyzed tracheal allografts for expression of components of the IL-13 signaling pathway by RT-PCR and immunocytochemical analyses. RESULTS: Interleukin-13 expression was detected in type II alveolar epithelial cells and human bronchial epithelial cells, but not in lung fibroblasts. IL-13 expression was decreased in whole lung and type II cells in response to alcohol exposure. In all cell types analyzed, expression of IL-13 signaling receptor (IL-13R alpha(1)) mRNA was markedly increased. In contrast, mRNA and protein expression of the IL-13 decoy receptor (IL-13R alpha(2)) were decreased in all cells analyzed. Exposure to alcohol also increased STAT6 phosphorylation in response to IL-13 and lipopolysaccharide. CONCLUSIONS: Data from multiple cell types in the pulmonary system suggest that IL-13 and its receptors play a role in alcohol-mediated activation of pro-fibrotic pathways. Taken together, these data suggest that alcohol primes the airway for increased IL-13 signaling and subsequent tissue remodeling upon injury such as transplantation. PMID- 19120069 TI - The role of eosinophil major basic protein in angiogenesis. AB - BACKGROUND: Eosinophil-derived major basic protein (MBP) plays an active role in allergic inflammation and tissue remodelling. However, its role in angiogenesis has not been established as yet. Therefore our objective was to investigate whether MBP exhibits any direct pro-angiogenic effects. METHODS: Rat aortic endothelial cells and human umbilical vascular endothelial cells were cultured with different concentrations of MBP and their viability (Trypan blue exclusion test), proliferation (thymidine incorporation) and capillary-like structure formation (matrigel assay) were investigated in vitro. The angiogenic activity of MBP was then tested in vivo using the chick chorio allantoic membrane (CAM) assay. RESULTS: Subcytotoxic concentrations of MBP induce endothelial cell proliferation and enhance the pro-mitogenic effect of vascular endothelial growth factor (VEGF), but do not affect their VEGF release. MBP promotes capillarogenesis by endothelial cells seeded on matrigel and sprouting formation in the CAM assay. Furthermore, we have shown that the pro-angiogenic effect of MBP is not due to its cationic charge since stimulation of the CAMs with the synthetic polycation, poly-L-arginine does not induce any angiogenic effects. CONCLUSIONS: These data demonstrate that MBP has pro-angiogenic effects in vitro and in vivo, providing a novel mechanism whereby MBP can participate in tissue inflammation and remodelling in atopic diseases. PMID- 19120070 TI - Cigarette smoke facilitates allergen penetration across respiratory epithelium. AB - BACKGROUND: The association between cigarette smoke exposure and allergic airway disease is a matter for debate. We sought to investigate in an in vitro system whether active smoking reduces the integrity and barrier function of the respiratory epithelium and thus facilitates allergen penetration. METHODS: We cultured the human bronchial epithelial cell line 16HBE14o- in a transwell culture system as a surrogate for the intact respiratory epithelium. The cell monolayer was exposed to standardized cigarette smoke extract (CSE). The extent and effects of trans-epithelial allergen penetration were measured using 125I labelled purified major respiratory allergens (rBet v 1, rPhl p 5 and rDer p 2) and histamine release experiments. RESULTS: Exposure of cells to concentrations of CSE similar to those found in smokers induced the development of para-cellular gaps and a decrease in trans-epithelial resistance. CSE exposure induced a more than threefold increase in allergen penetration. Increased subepithelial allergen concentrations provoked a substantial augmentation of histamine release from sensitized basophils. CONCLUSIONS: Our results indicate that cigarette smoke is a potent factor capable of reducing the barrier function of the respiratory epithelium for allergens and may contribute to increased allergic inflammation, exacerbation of allergic disease and boosting of IgE memory. PMID- 19120071 TI - Predictors of bronchial hyperresponsiveness in chronic rhinosinusitis with nasal polyp. AB - BACKGROUND: Chronic rhinosinusitis with nasal polyposis (CRSNP) and asthma are inflammatory lesions of the respiratory epithelium. This study was conducted to evaluate predictive factors of bronchial hyperresponsiveness (BHR) in patients with CRSNP. METHODS: BHR was evaluated using a methacholine bronchoprovocation test (MBPT) in 122 consecutive patients newly diagnosed with CRSNP at Seoul National University Hospital from January 2004 to June 2006. The following parameters were analyzed and compared between the BHR and non-BHR groups: symptoms, atopic status, current smoking, disease severity of CRSNP based on the Lund-Mackay scoring system of sinus CT, and counts of eosinophils in the serum and nasal tissues. RESULTS: Thirty-five percent of the patients were found to have BHR, and BHR was found to occur more frequently in patients that were currently suffering from sneezing (P = 0.007). In addition, the mean eosinophil counts of the serum and nasal tissues were higher in the BHR group than in the non-BHR group (P = 0.001 for the serum, P = 0.045 for the nasal tissues), and the eosinophil counts of the serum correlated to those of the nasal tissues (r = 0.334, P = 0.013). The disease severity, as determined by the Lund-Mackay scoring system, was not different between the two groups (P > 0.05). The best cutoff serum eosinophil count for predicting BHR in CRSNP patients was determined to be 300 cells/microl (sensitivity 70%, specificity 70%). CONCLUSION: Taken together, these results indicate that moderate to severe sneezing and a serum eosinophil count > or = 300 cells/microl may be predictive factors for BHR in patients with CRSNP. PMID- 19120072 TI - Immunomodulatory properties of Lactobacillus plantarum and its use as a recombinant vaccine against mite allergy. AB - BACKGROUND: Selected lactic acid bacteria were reported to prevent atopic dermatitis and experimental asthma but the mechanisms of their immunomodulatory effects are not fully elucidated. In this study, the signaling pathways triggered by Lactobacillus plantarum NCIMB8826 were investigated and the potential use of this strain producing a variant of the mite allergen Der p 1 as live vaccine vehicle was evaluated. METHODS: Mouse bone marrow-derived dendritic cells were stimulated with wild-type or a L. plantarum teichoic acid mutant to evaluate the secretion of cytokines. A recombinant L. plantarum expressing Der p 1 was engineered, its in vitro immunomodulatory properties were characterized and its prophylactic potential was evaluated in a Der p 1-sensitization murine model. RESULTS: Mouse dendritic cells stimulated by L. plantarum triggered the release of interleukin-10 (IL-10), IL-12 p40, IL-12 p70 and tumor necrosis factor-alpha (TNF-alpha). IL-12 p40 secretion was dependent on nuclear factor-kappaB (NF kappaB), mitogen-activated protein (MAP) kinases, Toll-like receptor 2 (TLR2), TLR9 and on the bacterial teichoic acid composition. Recombinant L. plantarum producing Der p 1 exhibited similar immunostimulatory properties as wild-type. Prophylactic intranasal pretreatment of mice with this recombinant strain prevented the development of the typical Th2-biased allergic response by a drastic reduction of specific IgE and the induction of protective allergen specific IgG2a antibodies. Moreover, both wild-type or recombinant L. plantarum reduced airway eosinophilia following aerosolized allergen exposure and IL-5 secretion upon allergen restimulation. CONCLUSION: By combining both Th1-type immunostimulatory properties and an efficient allergen delivery capacity, recombinant L. plantarum producing Der p 1 represents a promising vaccine against house dust mite allergy. PMID- 19120073 TI - Hymenoptera venom allergy: analysis of double positivity to honey bee and Vespula venom by estimation of IgE antibodies to species-specific major allergens Api m1 and Ves v5. AB - BACKGROUND: In patients with hymenoptera venom allergy diagnostic tests are often positive with honey bee and Vespula venom causing problems in selection of venoms for immunotherapy. METHODS: 100 patients each with allergic reactions to Vespula or honey bee stings and positive i.e. skin tests to the respective venom, were analysed for serum IgE to bee venom, Vespula venom and crossreacting carbohydrate determinants (CCDs) by UNICAP (CAP) and ADVIA Centaur (ADVIA). IgE-antibodies to species specific recombinant major allergens (SSMA) Api m1 for bee venom and Ves v5 for Vespula venom, were determined by ADVIA. 30 history and skin test negative patients served as controls. RESULTS: By CAP sensitivity was 1.0 for bee and 0.91 for Vespula venom, by ADVIA 0.99 for bee and 0.91 for Vespula venom. None of the controls were positive with either test. Double positivity was observed in 59% of allergic patients by CAP, in 32% by ADVIA. slgE to Api m1 was detected in 97% of bee and 17% of Vespula venom allergic patients, slgE to Ves v5 in 87% of Vespula and 17% of bee venom allergic patients. slgE to CCDs were present in 37% of all allergic patients and in 56% of those with double positivity and were more frequent in bee than in Vespula venom allergic patients. CONCLUSIONS: Double positivity of IgE to bee and Vespula venom is often caused by crossreactions, especially to CCDs. IgE to both Api m1 and Ves v5 indicates true double sensitization and immunotherapy with both venoms. PMID- 19120075 TI - Donor-specific immune regulation by CD8 lymphocytes expanded from rejecting human cardiac allografts. AB - To assess whether regulatory T cells are present in rejecting human cardiac allografts, we performed functional analyses of graft lymphocytes (GLs) expanded from endomyocardial biopsies (EMB; n = 5) with histological signs of acute cellular rejection. The GL cultures were tested for their proliferative capacity and regulatory activity on allogeneic-stimulated peripheral blood mononuclear cells (PBMC) of the patient (ratio PBMC:GLs = 5:1). Three of these GL cultures were hyporesponsive to donor antigens and suppressed the antidonor proliferative T-cell response of PBMC, but not the anti-third-party response. Interestingly, it was the CD8(+) GL subset of these cultures that inhibited the antidonor response (65-91% inhibition of the proportion of proliferating cells); the CD4(+) GLs of the expanded GL cultures were not suppressive. In conclusion, CD8(+) GLs expanded from rejecting human cardiac allografts can exhibit donor-specific immune regulatory activities in vitro. We suggest that during acute cellular rejection, GLs may not only consist of graft-destructing effector T cells, but also of cells of the CD8(+) type with the potential to specifically inhibit antidonor immune reactivity. PMID- 19120074 TI - An assessment of herpesvirus co-infections in patients with CMV disease: correlation with clinical and virologic outcomes. AB - The effect of herpesvirus co-infections (HHV-6, HHV-7) on cytomegalovirus (CMV) disease and its response to therapy is unknown. We prospectively analyzed herpesvirus co-infections in transplant recipients with CMV disease. All patients received 3 weeks of antiviral therapy. Samples were collected at baseline (day 0) and then day 3, 7, 14 and 21 poststart of therapy. Viral load testing for CMV, HHV-6 and HHV-7 was done using quantitative PCR assays in 302 patients of whom 256 had documented symptomatic CMV viremia. In this subset, day 0 HHV-6 co infection was present in 23/253 (9.1%) and HHV-7 in 17/253 (6.7%). Including those positive at any time point raised the prevalence to 79/256 (30.9%) for HHV 6 and 75/256 (29.3%) for HHV-7. Viral co-infection did not influence the response of CMV disease to antiviral therapy. Baseline CMV viral loads, time to eradication and risk of recurrence were similar in patients with and without HHV 6 or HHV-7 co-infection. Ganciclovir and valganciclovir had no clear effect on HHV-6 and HHV-7 viremia. In conclusion, herpesvirus co-infections are common in patients with CMV disease but with standard antiviral therapy, no clear clinical effects are discernable. Routine monitoring for viral co-infection in patients with CMV disease is not indicated. PMID- 19120076 TI - Plasma cytokines and chemokines in primary graft dysfunction post-lung transplantation. AB - Primary graft dysfunction (PGD) after lung transplantation causes significant morbidity and mortality. We aimed to determine the role of cytokines and chemokines in PGD. This is a multicenter case-control study of PGD in humans. A Luminex analysis was performed to determine plasma levels of 25 chemokines and cytokines before and at 6, 24, 48 and 72 h following allograft reperfusion in 25 cases (grade 3 PGD) and 25 controls (grade 0 PGD). Biomarker profiles were evaluated using a multivariable logistic regression and generalized estimating equations. PGD cases had higher levels of monocyte chemotactic protein-1 (MCP 1)/chemokine CC motif ligand 2 (CCL2) and interferon (IFN)-inducible protein (IP 10)/chemokine CXC motif ligand 10 (CXCL10) (both p < 0.05), suggesting recruitment of monocytes and effector T cells in PGD. In addition, PGD cases had lower levels of interleukin (IL-13) (p = 0.05) and higher levels of IL-2R (p = 0.05). Proinflammatory cytokines, including tumor necrosis factor (TNF)-alpha, and IFN-gamma decreased to very low levels after transplant in both PGD cases and controls, exhibiting no differences between the two groups. These findings were independent of clinical variables including diagnosis in multivariable analyses, but may be affected by cardiopulmonary bypass. Profound injury in clinical PGD is distinguished by the upregulation of selected chemokine pathways, which may useful for the prediction or early detection of PGD if confirmed in future studies. PMID- 19120077 TI - Delayed introduction of reduced-dose tacrolimus, and renal function in liver transplantation: the 'ReSpECT' study. AB - We report a multicenter, prospective, randomized, open-label trial investigating the effect of lower levels and delayed introduction of tacrolimus on renal function in liver transplant recipients. Adult patients with good renal function undergoing primary liver transplant were randomized to either: group A (standard dose tacrolimus [target trough levels >10 ng/mL] and corticosteroids; n = 183); group B (mycophenolate mofetil [MMF] 2g/day, reduced-dose tacrolimus [target trough levels /=60 and those with preoperative MELD >/=30. D-MELD >/=1600 accurately predicted worse outcome in recipients with and without hepatitis C. There is significant regional variation in average D-MELD scores at transplant, however, regions with larger numbers of high D-MELD matches do not have higher survival rates. D-MELD is a simple, highly predictive tool for estimating outcomes after liver transplantation. This statistic could assist surgeons and their patients in making organ acceptance decisions. Applying D-MELD to liver allocation could eliminate many donor/recipient matches likely to have inferior outcome. PMID- 19120080 TI - The clinical impact of an early decline in kidney function in patients following heart transplantation. AB - Renal dysfunction is a well-known complication following heart transplantation. We examined an early decline in kidney function as a predictor of progression to end-stage renal disease and mortality in heart transplant recipients. We performed a retrospective cohort study of 233 patients who received a heart transplant between July 1985 and July 2004, and who survived >1 month. The decline in estimated creatinine clearance (CrCl) was used to predict the outcomes of need for chronic dialysis or mortality >1-year posttransplant. The earliest time to chronic dialysis was 484 days. A 30% decline in CrCl between 1 month and 12 months predicted the need for chronic dialysis (p = 0.01), all-cause mortality (p < 0.0001) and time to first CrCl 1-year posttransplant (p = 0.02). A 30% decline in CrCl between 1 month and 3 months also independently predicted the need for chronic dialysis (p = 0.04) and time to first CrCl 1-year posttransplant (p = 0.01). In conclusion, an early drop in CrCl within the first year is a strong predictor of chronic dialysis and death >1-year postheart transplantation. Future studies should focus on kidney function preservation in those identified at high risk for progression to end-stage kidney disease and mortality. PMID- 19120081 TI - Transplant center volume and outcomes after liver retransplantation. AB - Liver retransplantation surgery has a high rate of allograft failure due to patient comorbidities and technical demands of the procedure. Success of liver retransplantation could depend on surgeon experience and processes of care that relate to center volume. We performed a retrospective cohort study of adult liver retransplantation procedures performed from January 1, 1996 through December 31, 2005 using registry data from the Organ Procurement Transplantation Network. The primary outcome was 1-year allograft failure. Liver transplant centers were categorized as small, intermediate or high volume by dividing overall liver transplants into three tertiles of approximately equal size. Mean annual volume of overall liver transplants was <50 for low-volume centers, 50-88 for intermediate-volume centers and >88 for high-volume centers. The primary analysis consisted of 3977 liver retransplantation patients. The unadjusted risk of 1-year allograft failure was 37.8%. In multivariable logistic regression, the risk of 1 year allograft failure was not significantly different between low- (reference), intermediate- (OR 0.86, CI 0.72-1.03, p = 0.11) and high-volume centers (OR 0.88, CI 0.74-1.04, p = 0.14). Results were similar when the analysis was limited to retransplantation performed >160 days after initial transplantation. Center volume is an imprecise surrogate measure for 1-year outcomes after liver retransplantation. PMID- 19120082 TI - Incidence and severity of acute cellular rejection in recipients undergoing adult living donor or deceased donor liver transplantation. AB - Living donor liver transplantation (LDLT) may have better immunological outcomes compared to deceased donor liver transplantation (DDLT). The aim of this study was to analyze the incidence of acute cellular rejection (ACR) after LDLT and DDLT. Data from the adult-to-adult living donor liver transplantation (A2ALL) retrospective cohort study on 593 liver transplants done between May 1998 and March 2004 were studied (380 LDLT; 213 DDLT). Median LDLT and DDLT follow-up was 778 and 713 days, respectively. Rates of clinically treated and biopsy-proven ACR were compared. There were 174 (46%) LDLT and 80 (38%) DDLT recipients with >/=1 clinically treated episodes of ACR, whereas 103 (27%) LDLT and 58 (27%) DDLT recipients had >/=1 biopsy-proven ACR episode. A higher proportion of LDLT recipients had clinically treated ACR (p = 0.052), but this difference was largely attributable to one center. There were similar proportions of biopsy proven rejection (p = 0.97) and graft loss due to rejection (p = 0.16). Longer cold ischemia time was associated with a higher rate of ACR in both groups despite much shorter median cold ischemia time in LDLT. These data do not show an immunological advantage for LDLT, and therefore do not support the application of unique posttransplant immunosuppression protocols for LDLT recipients. PMID- 19120083 TI - Predictive ability of pretransplant comorbidities to predict long-term graft loss and death. AB - Whether to include additional comorbidities beyond diabetes in future kidney allocation schemes is controversial. We investigated the predictive ability of multiple pretransplant comorbidities for graft and patient survival. We included first-kidney transplant deceased donor recipients if Medicare was the primary payer for at least one year pretransplant. We extracted pretransplant comorbidities from Medicare claims with the Clinical Classifications Software (CCS), Charlson and Elixhauser comorbidities and used Cox regressions for graft loss, death with function (DWF) and death. Four models were compared: (1) Organ Procurement Transplant Network (OPTN) recipient and donor factors, (2) OPTN + CCS, (3) OPTN + Charlson and (4) OPTN + Elixhauser. Patients were censored at 9 years or loss to follow-up. Predictive performance was evaluated with the c statistic. We examined 25 270 transplants between 1995 and 2002. For graft loss, the predictive value of all models was statistically and practically similar (Model 1: 0.61 [0.60 0.62], Model 2: 0.63 [0.62 0.64], Models 3 and 4: 0.62 [0.61 0.63]). For DWF and death, performance improved to 0.70 and was slightly better with the CCS. Pretransplant comorbidities derived from administrative claims did not identify factors not collected on OPTN that had a significant impact on graft outcome predictions. This has important implications for the revisions to the kidney allocation scheme. PMID- 19120084 TI - Belatacept as maintenance immunosuppression for postrenal transplant de novo drug induced thrombotic microangiopathy. AB - De novo posttransplant thrombotic microangiopathy (TMA) is a complication of solid organ transplantation, which remains difficult to treat. In many cases, immunosuppressants and particularly calcineurin inhibitors, trigger TMA. Although withdrawing the offending drug may lead to resolution of TMA, graft and patient outcomes are poor. Specific treatments, including plasma exchange, have not gained widespread acceptance in those with fulminant disease and new approaches to the condition are urgently needed. We report a case of posttransplant de novo TMA presenting serially in association with ciclosporin, tacrolimus and sirolimus in a young recipient of a living donor kidney transplant. We describe a patient treated with belatacept, a novel CTLA4 Ig fusion protein, as ongoing maintenance immunosuppression to allow avoidance of conventional agents once associated with TMA. We report excellent early graft outcome, with no adverse events using this strategy. We suggest that belatacept may have a role in this traditionally difficult-to-treat group of patients. PMID- 19120085 TI - Long-term insulin-independence after allogeneic islet transplantation for type 1 diabetes: over the 10-year mark. AB - Results of islet of Langerhans transplantation have markedly improved in recent years, but most patients still lose insulin independence in the long-term. We report herein the longest (over 11 years) case of insulin independence after allogeneic islet transplantation. The subject had a 27-year history of type 1 diabetes and received a single islet-after-kidney graft of 8800 islet equivalents (IEQ)/kg, pooled from 2 donors. Insulin was discontinued by 3 months posttransplant and the patient has remained off insulin ever since. Yearly follow up studies have revealed normal metabolic control, including normal oral glucose tolerance test (OGTT). Reasons for success may involve choice of immunosuppression, low metabolic demand and low immune responsiveness as suggested by an excellent HLA matching and a high count of circulating regulatory T cells. This observation is so far an exceptional case, but clearly demonstrates the validity of the concept that long-term insulin independence after allogeneic islet transplantation is an achievable target. PMID- 19120086 TI - Variation in neighbouring genes of the dopaminergic and serotonergic systems affects feather pecking behaviour of laying hens. AB - Feather pecking is a behavioural disorder of laying hens and has serious animal welfare and economic implications. One of the several aetiological hypotheses proposes that the disorder results from redirected exploratory behaviour. Variation in the gene encoding the dopamine D4 receptor (DRD4) has been shown to be associated with exploratory behaviour in several species, including in a passerine bird species. We therefore considered DRD4 as a candidate gene for feather pecking. We have annotated DRD4 in the chicken genome and have re sequenced it in 140 animals belonging to: experimental layer lines divergently selected for high and low propensity to feather pecking; the unselected founder population; and two commercial lines with low and high propensity to feather pecking. We have identified two sub-haplotypes of DRD4 that are highly significantly associated with feather pecking behaviour in the experimental (P = 7.30 x 10(-7)) as well as in the commercial lines (P = 2.78 x 10(-6)). Linkage disequilibrium (LD) extends into a neighbouring gene encoding deformed epidermal autoregulatory factor 1 (DEAF1). The product of DEAF1 regulates the transcription of the gene encoding the serotonin (5-hydroxytryptamine) 1A receptor. Thus, DEAF1 represents another candidate gene for feather pecking. Re-sequencing of five animals homozygous for the 'low-pecking' sub-haplotype and of six animals homozygous for the 'high-pecking' sub-haplotype delineated an LD block of 14 833 bases spanning the two genes. None of the variants in the LD block is obviously functional. However, the haplotype information will be useful to select against the propensity to feather pecking in chicken and to elucidate the functional implications of the variants. PMID- 19120087 TI - Stress, neurotransmitters, and hormones neuroendocrine and genetic mechanisms. Preface. PMID- 19120088 TI - Identifying the stress transcriptome in the adrenal medulla following acute and repeated immobilization. AB - Stress triggers changes in gene expression mediating important adaptive and maladaptive responses. The full repertoire of genes whose expression in the adrenal medulla is altered by stress has not been previously determined. In this study, gene profiling (RAE 230 2.0 Affymetrix) was applied to elucidate global changes in gene expression in adrenal medulla of rats exposed to 2-h immobilization (IMO) stress once or repeatedly for 6 consecutive days. The number of transcripts significantly (P < 0.01) altered with single IMO (651 up, 487 down) was more than with repeated IMO (370 up, 195 down). The annotated transcripts were further analyzed and categorized. The largest numbers of changes were in mRNA levels in the transcription factor and cell signaling categories. Robust changes were also observed in transcripts related to growth factors, apoptosis, neurosecretion/neuropeptides, heat shock proteins, structural proteins, chemokines, cytokines, metabolism/lipid-metabolism, and proteases. Many (>80%) were uniquely induced by single IMO. About half of transcripts changed by repeated IMO were also responsive to single IMO. Pathway analysis was applied to identify direct interactions and common targets among gene products altered by single and repeated IMO. In this paper, we briefly describe the most pronounced changes observed, with emphasis on those that may provide new insight into the common and distinct mechanisms whereby the adrenal medulla responses to a first encounter with stress compared to repeated exposure to the same stressor. PMID- 19120089 TI - From Hans Selye's discovery of biological stress to the identification of corticotropin-releasing factor signaling pathways: implication in stress-related functional bowel diseases. AB - Selye pioneered the concept of biological stress in 1936, culminating in the identification of the corticotropin-releasing factor (CRF) signaling pathways by Vale's group in the last two decades. The characterization of the 41 amino-acid CRF and other peptide members of the mammalian CRF family, urocortin 1, urocortin 2, and urocortin 3, and the cloning of CRF(1) and CRF(2) receptors, which display distinct affinity for CRF ligands, combined with the development of selective CRF receptor antagonists enable us to unravel the importance of CRF(1) receptor in the stress-related endocrine (activation of pituitary-adrenal axis), behavioral (anxiety/depression, altered feeding), autonomic (activation of sympathetic nervous system), and immune responses. The activation of CRF(1) receptors is also one of the key mechanisms through which various stressors impact the gut to stimulate colonic propulsive motor function and to induce hypersensitivity to colorectal distension as shown by the efficacy of the CRF(1) receptor antagonists in blunting these stress-related components. The importance of CRF(1) signaling pathway in the visceral response to stress in experimental animals provided new therapeutic approaches for treatment of functional bowel disorder such as irritable bowel syndrome, a multifactor functional disorder characterized by altered bowel habits and visceral pain, for which stress has been implicated in the pathophysiology and is associated with anxiety-depression in a subset of patients. PMID- 19120090 TI - Investigating the molecular basis of major depressive disorder etiology: a functional convergent genetic approach. AB - Genes play a major role in behavioral adaptation to challenging environmental stimuli, but the complexity of their contribution remains unclear. There is growing evidence linking disease phenotypes with genes on the one hand, and the genesis of stress-related disorders like major depression, as a result of exposure to stressful environmental pathogens on the other. Here we illustrate the convergent role of monoaminergic genes in regulating the underlying biological mechanisms of stress and the emotions. By reviewing data that support a role of monoaminergic and other related genes in environmental adaptation, we conclude by advocating the use of convergent approaches in examining the genetic modulation of disease phenotypes. PMID- 19120091 TI - Stress-induced activation of neurons in the ventromedial arcuate nucleus: a blood brain-CSF interface of the hypothalamus. AB - In response to a pain-related acute stress, the expression of c-fos protein (Fos), a marker of acute neuronal excitation, was investigated in the hypothalamus of rats. Few Fos-immunopositive cells were seen 15 min after a single subcutaneous injection of 4% formalin in the hypothalamus, but only in the paraventricular nucleus (PVN). Fifteen minutes later, a high number of parvocellular neurons of the PVN showed Fos expression. By 60 min after injection, strong immunoreactivity appeared in the arcuate nucleus, but the Fos positive neurons distributed almost exclusively in the ventromedial subdivision of the nucleus. Neurons in this part of the arcuate nucleus express mainly neuropeptide Y (NPY) that projects to the medial parvocellular subdivision of the PVN. It has been demonstrated by previous studies that this part of the arcuate nucleus receives blood partly from the anterior pituitary through the subependymal plexus of the median eminence, and that it establishes, together with the median eminence, a blood-brain barrier-free area in the medial basal hypothalamus. Since the PVN-projecting NPY neurons in the arcuate neurons are sensitive to alterations in circulating corticosterone levels, the existence of a possible short feedback route in the stress-activated hypothalamo-pituitary adrenocortical system is discussed. PMID- 19120092 TI - Limbic regulation of hypothalamo-pituitary-adrenocortical function during acute and chronic stress. AB - The hypothalamo-pituitary-adrenocortical (HPA) axis is responsible for initiation of glucocorticoid stress responses in all vertebrate animals. Activation of the axis is regulated by diverse afferent input to the hypothalamic paraventricular nucleus (PVN). This review discusses brain mechanisms subserving generation and inhibition of stress responses focusing on the contribution of the limbic system and highlighting recent conceptual advances regarding organization of stress response pathways in the brain. First, control of HPA axis responses to psychogenic stimuli is exerted by a complex neurocircuitry that involves oligosynaptic networks between limbic forebrain structures and the PVN. Second, individual stress-modulatory structures can have a heterogeneous impact on HPA axis responses, based on anatomical micro-organization and/or stimulus properties. Finally, HPA axis hyperactivity pursuant to chronic stress involves a substantial functional and perhaps anatomical reorganization of central stress integrative circuits. Overall, the data suggest that individual brain regions do not merely function as monolithic activators or inhibitors of the HPA axis and that network approaches need be taken to fully understand the nature of the neuroendocrine stress response. PMID- 19120093 TI - Stress-induced changes in tyrosine hydroxylase gene expression in rat hypothalamic paraventricular, periventricular, and dorsomedial nuclei. AB - Particular hypothalamic nuclei--the paraventricular (PVN), periventricular (PeV), and dorsomedial (DMN)--are important centers regulating neuroendocrine and autonomic systems during stress. The present study investigated gene expression of catecholamines (CA)-synthesizing enzymes and protein levels, especially of tyrosine hydroxylase (TH), in response to acute (120 min) and repeated (7 x 120 min) immobilization (IMO) stress. Immunohistochemical staining revealed many TH positive perikarya in the PeV, less in the DMN, and a few in the PVN. Analysis of microdissected tissue samples obtained from frozen brain sections by quantitative competitive PCR method revealed TH mRNA in the PVN, periventricular subdivision of the PVN (PePVN), and DMN. TH mRNA and TH protein levels were increased in all the nuclei investigated, in both acute and repeated stress conditions. Interruption of ascending CA pathways from the brain stem areas abolished TH gene expression in the PVN and DMN in response to stress. The present data suggest that cell groups expressing TH gene are present in all three hypothalamic nuclei studied and are highly activated by stress. The regulation of the majority of these TH-positive cells during stress is under modulatory control by brain stem or spinal cord areas. PMID- 19120094 TI - Serotonergic systems, anxiety, and affective disorder: focus on the dorsomedial part of the dorsal raphe nucleus. AB - Depressed suicide patients have elevated expression of neuronal tryptophan hydroxylase 2 (TPH2) mRNA and protein in midbrain serotonergic neurons, as well as increases in brain serotonin turnover. The mechanisms underlying these changes are uncertain, but increased TPH2 expression and serotonin turnover could result from genetic influences, adverse early life experiences, or acute stressful life events, all of which can alter serotonergic neurotransmission and have been implicated in determining vulnerability to major depression. Emerging evidence suggests that there are several different stress-related subsets of serotonergic neurons, each with a unique role in the integrated stress response. Here we review our current understanding of how genetic and environmental factors may influence TPH2 mRNA expression and serotonergic neurotransmission, focusing in particular on the dorsomedial part of the dorsal raphe nucleus. This subdivision of the dorsal raphe nucleus is selectively innervated by key forebrain structures implicated in regulation of anxiety states, it gives rise to projections to a distributed neural system mediating anxiety states, and serotonergic neurons within this subdivision are selectively activated by a number of stress- and anxiety-related stimuli. A better understanding of the anatomical and functional properties of specific stress- or anxiety-related serotonergic systems should aid our understanding of the neural mechanisms underlying the etiology of anxiety and affective disorders. PMID- 19120095 TI - Degeneration of the locus ceruleus noradrenergic neurons in the stress-induced depression of rats. AB - We produced a model of depression in rats which have been exposed to 2-weeks forced walking stress. Electron microscopic observation on the locus ceruleus (LC) cells of the model rats disclosed low dense areas, destroyed membranes, aggregation of intracellular organs, and increased microglia. Density of LC axon terminals in the frontal cortex stained with dopamine beta-hydroxylase antiserum and percentage of LC cells stained with horseradish peroxidase or activated by electrical stimulation antidromically were low in the model. These indices increased in the model treated with imipramine. These findings suggest that the LC noradrenergic neurons degenerate in depression, but regenerate in remission. PMID- 19120096 TI - Effects of estrogen on stress-induced activation of peptide neurons in PVN of ovariectomized rats. AB - Estrogen has been implicated in brain function related to stress responses. We investigated whether estrogen affects psychological stress-induced activation of peptide-containing or nitric oxide-producing neurons in the paraventricular hypothalamic nucleus (PVN) in ovariectomized (OVX) rats, both placebo-treated (OVX + Pla) and estrogen-treated (OVX + E2) by comparison of c-Fos expression using immunohistochemistry. Cage-switch stress increased activation in oxytocinergic neurons in the parvocellular PVN (pPVN) in OVX + Pla, but not in that of OVX + E2, rats. Moreover, the stress-induced activation in NADPH diaphorase-positive neurons in the pPVN was larger in the OVX + E2 than in the OVX + Pla group. These findings suggest that estrogen attenuates the activation of oxytocinergic neurons in the pPVN, at least in part via nitric oxide. PMID- 19120097 TI - Stress-induced intercellular communication remodeling in the rat adrenal medulla. AB - To understand the mechanisms by which a prolonged exposure to stress enhances catecholamine secretion, we examined the effects of 5-day cold exposure on cell cell communication pathways in the rat adrenal medulla. Upon stress, the neurosecretory tissue undergoes dramatic morphofunctional changes resulting in increased chromaffin cell excitability, upregulation of both chemical transmission at the splanchnic nerve terminal-chromaffin cell synapses and spreading of gap junction-permeant Lucifer yellow between cells. All these changes converge to improve the stimulus-secretion coupling efficiency within the adrenal medulla and subsequently to adapt catecholamine release to a sustained organism demand. PMID- 19120098 TI - Cross-talk between adrenal medulla and adrenal cortex in stress. AB - Response of the endocrine system to stress is characterized by the concomitant release of catecholamines from the adrenal medulla and of glucocorticoids from the adrenal cortex. Therefore, the activity of these two embryologically different endocrine organs that are united under one organ capsule has to be synchronized. The data summarized here indicate an involvement of intra-adrenal interactions in this coordination of the body's response to stress. PMID- 19120099 TI - Effects of acute and chronic stressors and CRF in rat and mouse tests for depression. AB - Depressive illness is frequently associated with life stress. Corticotropin releasing factor (CRF) is believed to be a key brain mediator of behaviors associated with stress, and abnormalities in the function of CRF have been associated with depression. Therefore, we have studied the effects of acute and chronic footshock and restraint in tests used in rodents to assess depression like activity and antidepressant effects: the forced swim test in rats and mice, and the tail suspension test in mice. We also tested the effect of intracerebroventricular (icv) CRF administration. The results were complex. In the forced swim test in rats, acute footshock and restraint reduced floating, whereas chronic footshock increased floating as did icv CRF. However, chronic restraint induced opposite effects, decreasing floating in the forced swim test. The results from mice were significantly different. In the forced swim test, acute footshock and restraint decreased floating, while chronic footshock increased floating, and chronic restraint decreased floating as it did in rats. However, icv CRF decreased floating. The results from the tail-suspension test paralleled those from the forced swim test except that chronic footshock tended to decrease the time spent immobile. Thus in rats, the behavioral effects of the chronic footshock in the forced swim test could be explained by a desensitization of the CRF systems, either decreased activation of CRF, or desensitization of its receptors. However, such an effect cannot explain the responses to restraint, nor the behavioral effects of chronic footshock and restraint in mice. PMID- 19120100 TI - Peripheral lipopolysaccharide administration affects the olfactory dopamine system in mice. AB - Peripheral administration of lipopolysaccharide (LPS) in an amount that produces acute stress has been found to affect the catecholamine systems in the brain. Acute peripheral LPS administration activated norepinephrine (NE) metabolism in the locus ceruleus (LC). Approximately 40% of murine LC neurons project to the olfactory bulb (OB) and the anterior olfactory nucleus (AON). Thus, we investigated the effects of a single intra-peritoneal (i.p.) LPS injection on catecholamine biosynthesis in the OB and AON in 8-week-old C3H/HeN male mice. In the AON, the content of dopamine (DA), but not that of NE, was highly increased 6 h after LPS injection. In the OB, the contents of DA and NE did not change; but within 2 h after a single i.p. LPS injection, the mRNA levels of IkappaB, TNF alpha, and TNF-alpha receptor type 1 were significantly enhanced. Almost all TNF alpha-immunoreactive cells in the OB of the LPS-injected mice were located in the granule cell layer, and unexpectedly, they were not microglia but astroglia. The number of TUNEL-positive cells identified exclusively in the granule cell layer was significantly increased at 24 h after LPS injection. Therefore, our data suggest that astroglia activated by peripherally injected LPS may release TNF alpha, which may trigger apoptosis in the granule cell layer in the OB. The increase in DA content in the AON and the production of TNF-alpha and apoptotic cells in the OB by acute peripheral LPS administration are not likely to be related. PMID- 19120101 TI - Central neuropeptide Y in anxiety- and stress-related behavior and in ethanol intake. AB - One of the most profound properties of central neuropeptide Y (NPY) is its anxiolytic, or anti-anxiety, effect. This has been demonstrated repeatedly in a number of animal models. In addition, stressors affect NPY expression in the central nervous system, with acute and repeated (chronic) stress having differential effects. Here, a brief summary of some work performed in our laboratory is presented that supports a role for NPY in regulation of stress responses and behaviors. PMID- 19120102 TI - Activity of brain stem groups of catecholaminergic cells in tumor-bearing rats: response to immobilization stress. AB - The aim of the present study was to investigate the activity of tyrosine hydroxylase (TH) immunopositive neurons, measured by Fos protein expression, in the hindbrain noradrenergic (NA) cell groups in animals exposed to visceral tumor growth for 28 days induced by intraperitoneal implantation of fibrosarcoma cells. We were also interested in determining whether brain stem NA neurons of tumor bearing and intact animals exhibit similarities in their response to a strong heterotypic stimulus-immobilization (IMO) stress. We found increased Fos expression in NA cells of the nucleus tractus solitarii (A2 cell group) and of the A1 cell group of tumor-bearing rats. However, Fos expression in other brain stem NA cell groups, including A5, locus ceruleus, and A7, were similar to control rats. The effect of 60 min of IMO was evident in both groups, but there were no differences between Fos expression in brain stem NA cell groups in control and tumor-bearing rats. This indicates that the sensitivity of NA cells in tumor-bearing animals was not altered by the IMO-induced stress challenge. However, whether the increased Fos expression in NA cells in tumor-bearing animals is a consequence of a specific visceral response activated by cancer development or just a nonspecific event accompanying the cancer progression requires further study. PMID- 19120103 TI - Serotonergic changes produced by repeated exposure to forced swimming: correlation with behavior. AB - Repeated forced swim resulted in a decrease in the concentrations of serotonin (5 HT) and its metabolite 5-hydroxyindoleacetic acid in the hypothalamus and amygdala 24 h after the second swim session. This stressor also increased the mRNA levels for tryptophan hydroxylase-2, the rate-limiting enzyme in neuronal 5 HT synthesis, and 5-HT transporter in the midbrain as well as 5-HT1A receptor in the frontal cortex. Some of these serotonergic changes may be involved in the mechanisms of a depressive-like behavior induced by a stress of repeated swim in these animals. PMID- 19120104 TI - Activity of oxytocinergic neurons in the supraoptic nucleus under stimulation of alpha2-adrenoceptors in Brattleboro rats. AB - Vasopressin (AVP) deficient homozygous Brattleboro rats exhibit severe osmotic challenges due to waterless chronic hypernatremia and hyperosmolality. We investigated the effect of xylazine, an alpha2-adrenoceptor agonist, on the activity of oxytocinergic (OXY) neurons in the supraoptic nucleus (SON) of homozygous (di/di), heterozygous (di/+), and control (+/+) rats. Ninety minutes after saline (0.1 mL/100 g b.w., i.p.) or xylazine injection (10 mg/kg, i.p.) rats were anaesthetized with pentobarbital (50 mg/kg i.p.) and sacrificed by transcardial perfusion with fixative. Activity of OXY neurons was evidenced by nuclear Fos protein immunoreactivity. Fos/OXY colabelings were analyzed on 40-mum thick coronal sections using computerized light microscope. As expected, plasma osmolality and water intake revealed high heterogenity within the di/di group of rats. Fos expression in SON of di/di rats was correlated with osmolality of each rat. In saline-treated rats, maximum activation of Fos reached around 4% in +/+, 20% in di/+ rats, and as much as 60% in di/di rats. Xylazine activated in SON about 70% of OXY-ergic neurons in +/+, 60% in di/+ rats, and more than 80% in di/di rats. The present findings indicate that in spite of the high spontaneous activity of SON OXY-ergic neurons due to the AVP deficiency in di/di rats, many of the silent OXY-ergic neurons in the SON remained acceptable for alpha2 adrenoceptor stimulation. PMID- 19120105 TI - Changes in c-Fos and NOS expression in the PVH of lactating rats in response to excitotoxicity and stress. AB - This study investigated how lactation modified the expression patterns of Fos and nitric oxide synthase in the hypothalamic paraventricular nucleus (PVH) induced by excitotoxicity and stress. Kainic acid or egg white treatment weakly activated Fos expression in the PVH of lactating in comparison to diestrus or ovariectomized (OVX) rats. Labels for NADPH-diaphorase and nNOS revealed a different distribution pattern in the PVH depending on the physiological condition and challenge. The present results confirm that lactation attenuates the PVH activational response to stress and excitotoxicity, and both stimuli induced nitric oxide expression in the PVH of diestrus, lactating, and OVX rats. PMID- 19120106 TI - Exposure to severe stressors causes long-lasting dysregulation of resting and stress-induced activation of the hypothalamic-pituitary-adrenal axis. AB - Exposure to some predominantly emotional (electric shock) and systemic (interleukin-1beta) stressors has been found to induce long-term sensitization of the hypothalamic-pituitary-adrenal (HPA) responsiveness to further superimposed stressors. Since exposure to immobilization on wooden boards (IMO) is a severe stressor and may have interest regarding putative animal models of post-traumatic stress disorders (PTSD), we have characterized long-lasting effects of a single exposure to IMO and other stressors on the HPA response to the same (homotypic) and to novel (heterotypic) stressors and the putative mechanisms involved. A single exposure to IMO caused a long-lasting reduction of peripheral and central responses of the HPA axis, likely to be mediated by some brain areas, such as the lateral septum and the medial amygdala. This desensitization is not explained by changes in negative glucocorticoid feedback, and, surprisingly, it is positively related to the intensity of the stressors. In contrast, the HPA response to heterotypic stressors (novel environments) was enhanced, with maximal sensitization on the day after IMO. Sensitization progressively vanished over the course of 1-2 weeks and was not modulated by IMO-induced corticosterone release. Moreover, it could not be explained by changes in the sensitivity of the HPA axis to fast or intermediate/delayed negative feedback, as evaluated 1 week after exposure to IMO, using shock as the heterotypic stressor. Long-lasting stress induced behavioral changes reminiscent of enhanced anxiety and HPA sensitization are likely to be parallel but partially independent phenomena, the former being apparently not related to the intensity of stressors. PMID- 19120107 TI - Chronic stress and individual vulnerability. AB - Over the last decades the burden of disease in Western countries has shifted from comparably easily treated infectious diseases to more complex diseases, such as the metabolic syndrome, cardiovascular disease, and psychiatric disorders. A common characteristic of these illnesses is the interplay of multiple genetic and nongenetic factors, which eventually results in the manifestation of disease symptoms. Large-scale epidemiological studies in humans have resulted in the identification of various environmental and genetic risk factors, which contribute to the onset, duration, and severity of disease. While tremendous progress has been made, it is still impossible to predict which combination of risk factors will result in the manifestation of a specific illness. This lack of knowledge is also frequently reflected in inadequate treatment strategies, which mainly focus on symptom reversal rather than targeting the cause of the diseases. One of the most prominent environmental risk factors described for numerous diseases is chronic exposure to stressful situations. In this paper we address clinical and preclinical evidence of chronic stress as a risk factor for disease and introduce a novel, high-throughput mouse model for chronic social stress. We can show that this model has a high degree of construct, face, and predictive validity in terms of physiological, behavioral, and gene expression changes. We further illustrate how novel animal models of chronic social stress can help to unravel the complex interaction of individual genetic vulnerability and environmental risk factors. PMID- 19120108 TI - Defensive behavioral strategies and enhanced state anxiety during chronic subordinate colony housing are accompanied by reduced hypothalamic vasopressin, but not oxytocin, expression. AB - Chronic subordinate colony (CSC) housing has recently been shown to be a clinically relevant model of chronic psychosocial stress for male mice and to cause an increase in the animals' anxiety-related behavior on the plus-maze. Here, we investigated (1) the detailed subordinate/dominant behavior during CSC housing, (2) the anxiety-related behavior of CSC and control mice in two independent tests, and (3) whether CSC exposure also influences the hypothalamic expression of oxytocin (OXT) and/or arginine vasopressin (AVP), specifically within the paraventricular nucleus. Both neuropeptides are known to be involved in the regulation of anxiety and stress responses. Behavioral observation revealed that all male CSC mice that were co-housed with a slightly larger male mouse obtained a subordinate status within their colonies over 19 consecutive days. Furthermore, CSC exposure resulted in diminished body weight gain and increased anxiety-related behavior as quantified both on the elevated plus-maze and in the light-dark box. Hypothalamic mRNA levels of OXT remained unchanged, whereas AVP mRNA was found to be decreased on day 20 of CSC exposure. In conclusion, exposure to CSC enhances anxiety, an effect that seems to be independent of the hypothalamic expression of the neuropeptides OXT and AVP. PMID- 19120109 TI - The response of plasma catecholamines in rats simultaneously exposed to immobilization and painful stimuli. AB - Immobilization represents a strong stressor inducing a profound increase in plasma epinephrine and norepinephrine levels. We have previously demonstrated that a subcutaneous injection of formalin (0.2 mL of 4% solution/100 g bw) attenuated the immobilization-induced elevation of plasma epinephrine levels in rats. In the present study, we investigated whether other painful and stressful stimuli, such as capsaicin, hydrochloric acid, mechanical pressure, heat, and cold, might also attenuate the increase of plasma epinephrine in rats exposed to acute immobilization stress. With the exception of formalin, all of the painful stimuli applied failed to attenuate the increase of plasma epinephrine levels in immobilized animals. Our data suggest that the attenuation of an immobilization induced increase in plasma epinephrine levels is specific for subcutaneous formalin administration. PMID- 19120110 TI - Hypergravity-induced increase in plasma catecholamine and corticosterone levels in telemetrically collected blood of rats during centrifugation. AB - Rats subjected to various accelerations (+G) exhibited increased levels of plasma epinephrine (EPI), norepinephrine (NE), and corticosterone. However, the collection of blood was performed after a centrifugation finished, and therefore the levels could be affected by the process of deceleration. The aim of this study was to evaluate plasma EPI, NE, and corticosterone levels in blood collected directly during centrifugation after reaching different G (2-6), using newly developed remote-controlled equipment. Animals placed into the centrifuge cabins had inserted polyethylene tubing in the tail artery, which was connected with a preprogrammed device for blood withdrawals. Plasma EPI, NE, and corticosterone levels were measured at different time intervals of hypergravity of 2-6G. Plasma EPI levels showed a huge, hypergravity-level-dependent increase. After the last blood collection was completed during hypergravity, the centrifuge was turned off and another blood sampling was performed immediately after the centrifuge stopped (10 min). In these samples, plasma EPI showed significantly lower levels compared to centrifugation intervals. Plasma NE levels were significantly increased after 6G only. The increase in plasma corticosterone was dependent on level of G, however after the centrifuge stopped, corticosterone levels remained elevated. Thus, our data show that hypergravity highly activates the adrenomedullary and hypothalamo-pituitary-adrenocortical systems, whereas the sympathoneural system is activated only at high hypergravity. Immediately after centrifugation is over, EPI levels quickly return to control values. Our technique of blood collection during centrifugation allows assessment of the real hormonal levels at the particular hypergravity value. PMID- 19120111 TI - Contribution of glucocorticoids to protective influence of preconditioning mild stress against stress-induced gastric erosions. AB - It is known that preconditioning stress may attenuate stress-induced gastric injury and that this effect is mediated by prostaglandins. In the present study we investigated the contribution of glucocorticoids to the gastroprotective effect of preconditioning stress. The effects of mild stress on gastric erosion caused by severe stress were compared in rats with normal and deficient corticosterone response to preconditioning mild stress. Mild stress decreased the gastric ulceration caused by severe stress, and this effect was prevented by glucocorticoid deficiency during mild stress. The results suggest that glucocorticoids released during preconditioning mild stress contribute to the gastroprotective effect of this stress. PMID- 19120112 TI - Estrogen replacement suppresses pressor response and oxidative stress induced by cage-switch stress in ovariectomized rats. AB - We examined the suppressive effects of estradiol on psychological stress-induced cardiovascular responses and oxidative stress in ovariectomized rats, both placebo-treated (OVX+Pla) and estrogen-treated (OVX+E2). The elevations in blood pressure and heart rate induced by cage-switch stress were attenuated in the OVX+E2 as compared with the OVX+Pla group. N(G)-nitro-L-arginine methyl ester, administered via drinking water, reduced the difference in these responses. Furthermore, this stress increased plasma nitrotyrosine and decreased plasma nitric oxide (NO) metabolites only in the OVX+Pla group. We demonstrated that estrogen replacement suppresses cardiovascular responses to psychological stress, at least in part by improving NO bioavailability in ovariectomized rats. PMID- 19120113 TI - Psychosocial stress and volatile compounds in preputial glands of rats. AB - Pheromones are volatile chemical cues that in rodents originate in large part from the preputial glands and influence various behavioral and physiological processes. We have examined the effect of psychosocial stress on volatile compound composition of preputial glands of male rats. Rats were housed three per cage or singly for 70 days. Dominants had heavier preputials compared to subdominant and subordinate rats. Capillary gas chromatographic mass spectrometry identified 55 volatile preputial compounds: 17 did not differ between groups, while 26 compounds were higher in single-housed compared to triad-housed rats. Six compounds were higher in dominant, while another six were higher in both dominant and single-housed rats. We conclude that both housing condition and rank status have significant differential effects on the preputial volatile compounds. PMID- 19120114 TI - Computer models of stress, allostasis, and acute and chronic diseases. AB - The past century has seen a profound shift in diseases of humankind. Acute, unifactorial diseases are being replaced increasingly by multifactorial disorders that arise from complex interactions among genes, environment, concurrent morbidities and treatments, and time. According to the concept of allostasis, there is no single, ideal set of steady-state conditions in life. Allostasis reflects active, adaptive processes that maintain apparent steady states, via multiple, interacting effectors regulated by homeostatic comparators- "homeostats." Stress can be defined as a condition or state in which a sensed discrepancy between afferent information and a set point for response leads to activation of effectors, reducing the discrepancy. "Allostatic load" refers to the consequences of sustained or repeated activation of mediators of allostasis. From the analogy of a home temperature control system, the temperature can be maintained at any of a variety of levels (allostatic states) by multiple means (effectors), regulated by a comparator thermostat (homeostat). Stress might exert adverse health consequences via allostatic load. This presentation describes models of homeostatic systems that incorporate negative feedback regulation, multiple effectors, effector sharing, environmental influences, intrinsic obsolescence, and destabilizing positive feedback loops. These models can be used to predict effects of environmental and genetic alterations on allostatic load and therefore on the development of multisystem disorders and failures. PMID- 19120115 TI - Chronic stress, combined with a high-fat/high-sugar diet, shifts sympathetic signaling toward neuropeptide Y and leads to obesity and the metabolic syndrome. AB - In response to stress, some people lose while others gain weight. This is believed to be due to either increased beta-adrenergic activation, the body's main fat-burning mechanism, or increased intake of sugar- and fat-rich "comfort foods." A high-fat, high-sugar (HFS) diet alone, however, cannot account for the epidemic of obesity, and chronic stress alone tends to lower adiposity in mice. Here we discuss how chronic stress, when combined with an HFS diet, leads to abdominal obesity by releasing a sympathetic neurotransmitter, neuropeptide Y (NPY), directly into the adipose tissue. In vitro, when "stressed" with dexamethasone, sympathetic neurons shift toward expressing more NPY, which stimulates endothelial cell (angiogenesis) and preadipocyte proliferation, differentiation, and lipid-filling (adipogenesis) by activating the same NPY-Y2 receptors (Y2Rs). In vivo, chronic stress, consisting of cold water or aggression in HFS-fed mice, stimulates the release of NPY and the expression of Y2Rs in visceral fat, increasing its growth by 50% in 2 weeks. After 3 months, this results in metabolic syndrome-like symptoms with abdominal obesity, inflammation, hyperlipidemia, hyperinsulinemia, glucose intolerance, hepatic steatosis, and hypertension. Remarkably, local intra-fat Y2R inhibition pharmacologically or via adenoviral Y2R knock-down reverses or prevents fat accumulation and metabolic complications. These studies demonstrated for the first time that chronic stress, via the NPY-Y2R pathway, amplifies and accelerates diet-induced obesity and the metabolic syndrome. Our findings also suggest the use of local administration of Y2R antagonists for treatment of obesity and NPY-Y2 agonists for fat augmentation in other clinical applications. PMID- 19120116 TI - Post-transcriptional regulation of tyrosine hydroxylase expression in adrenal medulla and brain. AB - It is well established that long-term stress leads to induction of tyrosine hydroxylase (TH) mRNA and TH protein in adrenal medulla and brain. This induction is usually associated with stimulation of the TH gene transcription rate. However, a number of studies have reported major discrepancies between the stress induced changes in TH gene transcription, TH mRNA, and TH protein. These discrepancies suggest that post-transcriptional mechanisms also play an important role in regulating TH expression in response to stress and other stimuli. In this report, we summarize some of our findings and literature reports that demonstrate these discrepancies in adrenal medulla, locus ceruleus, and midbrain dopamine neurons. We then describe our recent work investigating the molecular mechanisms that mediate this post-transcriptional regulation in adrenal medulla and midbrain. Our results suggest that trans-acting factors binding to the polypyrimidine-rich region of the 3' untranslated region of TH mRNA play a role in these post-transcriptional mechanisms. A hypothetical cellular model describing this post-transcriptional regulation is proposed. PMID- 19120117 TI - Adrenergic responses to stress: transcriptional and post-transcriptional changes. AB - Stress effects on adrenergic responses in rats were examined in adrenal medulla, the primary source of circulating epinephrine (Epi). Irrespective of duration, immobilization (IMMO) increased adrenal corticosterone to the same extent. In contrast, Epi changed little, suggesting that Epi synthesis replenishes adrenal pools and sustains circulating levels for the heightened alertness and physiological changes required of the "flight or fight" response. IMMO also induced the Epi-synthesizing enzyme, phenylethanolamine N-methyltransferase (PNMT). The rise in its mRNA and protein was preceded by increases in Egr-1 and Sp1 mRNA, protein, and protein-DNA binding complex formation. With repeated and prolonged stress, PNMT protein did not reflect the magnitude of change in mRNA. The latter suggests that post-transcriptional, in addition to transcriptional mechanisms, regulate PNMT responses to stress. To further reveal molecular mechanisms underlying stress-induced changes in adrenergic function, the effects of hypoxia on PNMT promoter-driven gene expression are being examined in adrenal medulla-derived PC12 cells. Hypoxia activates the PNMT promoter to increase PNMT promoter-driven luciferase reporter gene expression and endogenous PNMT in PC12 cells. Induction of both appear mediated via activation of multiple signaling pathways and downstream activation of hypoxia inducible factor and PNMT transcriptional activators, Egr-1 and Sp1. Hypoxia generates both partially and fully processed forms of PNMT mRNA. The former reportedly is translated into a truncated, nonfunctional protein, and the latter into enzymatically active PNMT. Together, findings suggest that stress increases PNMT gene transcriptional activity but post-transcriptional regulatory mechanisms limit the biological end point of functional PNMT enzyme and, thereby, Epi. PMID- 19120118 TI - Regulation of gene expression of catecholamine biosynthetic enzymes in dopamine beta-hydroxylase- and CRH-knockout mice exposed to stress. AB - Norepinephrine-deficient mice harbor a disruption of the gene for dopamine-beta hydroxylase (DBH-KO). Corticotropin-releasing hormone knockout mice (CRH-KO) have markedly reduced HPA activity. The aim of the present work was to study how deficiency of DBH and CRH would affect tyrosine hydroxylase (TH), DBH, and phenylethanolamine N-methyltransferase (PNMT) gene expression and protein levels in the adrenal medulla (AM) and stellate ganglia (SG) of control and stressed mice. Both in AM and SG, single immobilization significantly increased TH and DBH mRNA and protein levels both in wild-type (WT) and CRH-KO mice. On the other hand, the stress-triggered increase in PNMT mRNA and protein levels seen in WT mice was absent in CRH-KO mice. DBH-KO mice are more sensitive to stress but survive a single 2 h restraint stress in a tube. The increase in TH mRNA levels induced by restraint stress in WT was not observed in DBH-KO mice. PNMT mRNA and especially PNMT protein levels were significantly elevated in AM of DBH-KO mice. In SG of DBH-KO mice, TH mRNA levels were not affected; however, PNMT gene expression was highly elevated. Thus, disruption of the DBH gene surprisingly blocks the stress-induced elevation of TH mRNA levels in AM but increases PNMT gene expression in both AM and SG. Our data indicate that adrenergic signaling is required for stress-induced increase in TH mRNA and that this signaling restrains stress-induced increase in PNMT mRNA. They also confirm that the HPA system plays a crucial role in the stress-induced regulation of PNMT gene expression. PMID- 19120119 TI - Stress-induced alternative splicing modulations in brain and periphery: acetylcholinesterase as a case study. AB - Mammalian stress responses present a case study for investigating alternative splicing reactions in general, and changes in acetylcholinesterase (AChE) gene expression in particular, under endangered homeostasis. Acetylcholine (ACh) is a major regulator of stress responses, which was recently found to function as an essential route by which neurons can "talk" to immune cells. Therefore, chemical, physical, or psychological insults to the brain might all be traced in peripheral immune cells, which serve as key determinants in the physiological reactions to stress. Stress-induced changes in the alternative splicing patterns of AChE pre mRNA give this gene and its different protein products diverse stress responsive functions that are associated with both the enzymatic and noncatalytic properties of AChE variants. Transgenic manipulations of AChE gene expression uncovered previously nonperceived aspects of stress responses, including brain-to-blood as well as immune-to-neuronal communication. Herein we discuss the newly gained understanding achieved by using genomic manipulations of AChE gene expression as tools for approaching the alternative splicing features of mammalian stress responses. PMID- 19120121 TI - Impaired chronotropic response to exercise in mice lacking catecholamines in adrenergic cells. AB - To define the in vivo role of adrenergic catecholamines (CAs), we generated a mouse model whereby tyrosine hydroxylase (TH) was knocked out (KO) in phenylethanolamine N-methyltransferase-expressing cells. These adrenergic specific TH-KO mice were viable and grossly normal. Their resting heart rate and blood pressure, as monitored by telemetry, were unchanged. However, when challenged with treadmill exercise, their chronotropic responses were significantly reduced by 14% compared to wild-type mice. Thus, our data suggest that adrenergic CA is required for normal chronotropic responses to stress, but not required for prenatal and postnatal development or normal cardiovascular function at rest. PMID- 19120122 TI - Phenylethanolamine N-methyltransferase gene expression in the heart and blood pressure response to oxytocin treatment in rats exposed to voluntary wheel running. AB - This study aimed to contribute to the knowledge of cardiovascular regulation associated with repeated exercise by evaluating untraditional parameters in the model of voluntary wheel running. Possible changes in cardiac phenylethanolamine N-methyltransferase (PNMT) gene expression were evaluated using running for 3 weeks in four rat strains, and the hypothesis that voluntary wheel running modifies mean arterial pressure (MAP) responses to oxytocin administration was verified. Running activity increased gradually and was high in spontaneously hypertensive rats (SHR) and Sprague-Dawley rats, while low in Wistar rats. The levels of PNMT mRNA in the left but not right atrium increased significantly in rat strains exhibiting high physical activity. Concentrations of PNMT mRNA were significantly higher in SHR and Sprague-Dawley compared to those in Wistar rats, which ran much shorter distances. MAP was found to be higher in rats exposed to voluntary running, which might be the result of the cessation of the exercise 24 h before the measurements. Oxytocin treatment (5 microg/kg and 30 microg/kg i.v.) resulted in significant increase in MAP in both control and running animals in a dose-dependent manner. In conclusion, voluntary wheel running failed to modify sensitivity to cardiovascular action of oxytocin but resulted in increased gene expression of PNMT in left, but not right, heart atrium in a running activity dependent manner. PMID- 19120120 TI - Adrenergic polymorphism and the human stress response. AB - Tyrosine hydroxylase (TH) is the rate-limiting enzyme in catecholamine biosynthesis. Does common genetic variation at human TH alter autonomic activity and predispose to cardiovascular disease? We undertook systematic polymorphism discovery at the TH locus, and then tested variants for contributions to sympathetic function and blood pressure. We resequenced 80 ethnically diverse individuals across the TH locus. One hundred seventy-two twin pairs were evaluated for sympathetic traits, including catecholamine production and environmental (cold) stress responses. To evaluate hypertension, we genotyped subjects selected from the most extreme diastolic blood pressure percentiles in the population. Human TH promoter haplotype/reporter plasmids were transfected into chromaffin cells. Forty-nine single nucleotide polymorphisms (SNPs) and one tetranucleotide repeat were discovered, but coding region polymorphism did not account for common phenotypic variation. A block of linkage disequilibrium spanned four common variants in the proximal promoter. Catecholamine secretory traits were significantly heritable, as were stress-induced blood pressure changes. In the TH promoter, significant associations were found for urinary catecholamine excretion, as well as blood pressure response to stress. TH promoter haplotype #2 (TGGG) showed pleiotropy, increasing both norepinephrine excretion and blood pressure during stress. In hypertension, a case-control study (1266 subjects, 53% women) established the effect of C-824T in determination of blood pressure. We conclude that human catecholamine secretory traits are heritable, displaying joint genetic determination (pleiotropy) with autonomic activity and finally with blood pressure in the population. Catecholamine secretion is influenced by genetic variation in the adrenergic pathway encoding catecholamine synthesis, especially at the classically rate-limiting step, TH. The results suggest novel pathophysiological links between a key adrenergic locus, catecholamine metabolism, and blood pressure, and suggest new strategies to approach the mechanism, diagnosis, and treatment of systemic hypertension. PMID- 19120123 TI - Endocrine rhythms and expression of selected genes in the brain, stellate ganglia, and adrenals of hypertensive TGR rats. AB - Hypertensive TGR(mRen2)27 (TGR) rats represent a strain with genetically upregulated renin-angiotensin-aldosterone system. Simultaneously with development of hypertension, a daily profile in blood pressure (BP) inverts and in mature TGR rats BP is higher during the lighttime (L) than the darktime (D). Physiological mechanisms of inverted BP rhythm generation are not understood. In our study we determined circadian profiles of plasma hormones related to BP control (aldosterone, corticosterone, melatonin, prolactin) in TGR and control Sprague Dawley (SD) rats over 24 h and expression of genes encoding catecholamine synthesizing enzymes, tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT) in adrenals and stellate ganglia. Plasma levels of corticosterone and aldosterone were higher in TGR than SD rats but acrophases of their rhythms were not changed. Darktime peak of prolactin in TGR rats was decreased in comparison with SD animals and pineal melatonin levels started to rise earlier in TGR than in SD rats. In adrenals we found upregulated expression of TH, DBH, and PNMT mRNA at the beginning of the lighttime in TGR compared to SD rats. Expression of TH and DBH in stellate ganglia was not different in TGR rats in comparison with SD, but PNMT expression was higher during L compared to D in TGR rats. We hypothesize that upregulated adrenal medulla functioning in the morning and disturbed communication between circadian oscillators and mechanisms involved in BP control can explain the reversed BP profile in TGR rats. PMID- 19120124 TI - Catecholamine-synthesizing cells in the embryonic mouse heart. AB - The heart is a primary source of epinephrine and norepinephrine during embryonic development, yet little is known about the cardiac cells that produce these catecholamine hormones. To identify when and where catecholamine-synthesizing cells are found in the embryonic heart, we developed a novel mouse genetic model by "knocking-in" the Cre-recombinase gene to the locus encoding for the epinephrine biosynthetic enzyme, phenylethanolamine n-methyltransferase. When crossed with ROSA26 reporter mice, the beta-galactosidase gene is activated in adrenergic cells. A major advantage of this approach is that it allows detection of adrenergic cells and their progeny, regardless of whether the progeny cells retain an adrenergic phenotype or not. Our data show that adrenergic cells appear as early as embryonic day 8.5 and continue to accumulate in substantial numbers through birth in the mouse heart, where they appear to share common ancestry with myocardial lineages. Large numbers of atrial and especially ventricular myocytes appear to be derived from embryonic adrenergic cells in the heart. In addition, many of the pacemaking cells in the sinoatrial and atrioventricular nodes also appear to be derived from an adrenergic lineage. Thus, our results suggest that catecholamine-synthesizing cells serve as cardiomyocyte progenitors in the embryonic heart. PMID- 19120125 TI - Epinephrine regulation of hemodynamics in catecholamine knockouts and the pithed mouse. AB - Phenylethanolamine N-methyltransferase (PNMT) catalyzes synthesis of epinephrine (E) and is present in the brain, heart, and adrenal. E is a neurotransmitter and important hormone; however, its role in regulating cardiovascular dynamics is still unclear. We generated an E-deficient mouse model by knocking out the PNMT gene. The PNMT KO mouse had normal resting blood pressure, while treadmill exercise caused hypertension, suggesting an impaired response to stress in the absence of the stress hormone E. As PNMT occurs at a lower concentration in many extra-adrenal tissues including the brain, we set up a pithed mouse model to study the peripheral effects of E on cardiovascular dynamics, using pithing to eliminate central and reflex effects. The pithed mouse requires different surgical techniques and stimulation voltages than rats, and showed voltage- and frequency-dependent blood pressure responses to electrical stimuli. Stimulation with the alpha-adrenergic agonist phenylephrine gave a marked systolic pressor response, while the beta2 agonist salbutamol lowered diastolic blood pressure. The pithed PNMT KO mouse had an exaggerated blood pressure response to salbutamol, suggesting beta2 receptor supersensitivity. A targeted KO of tyrosine hydroxylase in PNMT-producing cells produced a mouse deficient in catecholamines in the adrenal. These targeted KO mice displayed significantly smaller pressor responses than pithed control mice. We find that E release during stress prevents an excessive increase in blood pressure. PMID- 19120126 TI - Type 1 and 2 IP3 receptors respond differently to catecholamines and stress. AB - Type 1 and 2 inositol 1,4,5-trisphosphate (IP3) receptors have been found in cardiac tissue, although they are localized in different types of cells. While the type 1 predominates in neuronal cells and cardiac ganglia, type 2 IP3 receptor is localized mainly in cardiomyocytes. In the heart, gene expression of the type 1 IP3 receptor is modulated by catecholamines, while type 2 is not affected. In this study we compared the effects of two stressors--cold exposure and hypoxia--on gene expression and protein levels of type 1 and 2 IP3 receptors. We found that both types of IP3 receptors were increased by these stressors, although the response of the type 2 IP3 receptor was delayed. We propose that regulation of these types of IP3 receptors in the heart might differ in control conditions and during stress. PMID- 19120127 TI - Human sympathetic nerve biology: parallel influences of stress and epigenetics in essential hypertension and panic disorder. AB - Patients with panic disorder provide a clinical model of stress. On a "good day," free from a panic attack, they show persistent stress-related changes in sympathetic nerve biology, including abnormal sympathetic nerve single-fiber firing ("salvos" of multiple firing within a cardiac cycle) and release of epinephrine as a cotransmitter. The coreleased epinephrine perhaps originates from in situ synthesis by phenylethanolamine N-methyltransferase (PNMT). In searching for biological evidence that essential hypertension is caused by mental stress--a disputed proposition--we note parallels with panic disorder, which provides an explicit clinical model of stress: (1) There is clinical comorbidity; panic disorder prevalence is increased threefold in essential hypertension. (2) For both, epinephrine cotransmission is present in sympathetic nerves. (3) In panic disorder and essential hypertension, but not in health, single-fiber sympathetic nerve firing salvos occur. (4) Tissue nerve growth factor is increased in both conditions (nerve growth factor is a stress reactant). (5) There is induction of PNMT in sympathetic nerves. Essential hypertension exhibits a further manifestation of mental stress: there is activation of noradrenergic brain stem neurons projecting to the hypothalamus and amygdala. These pathophysiological findings strongly support the view that chronic mental stress is important in the pathogenesis of essential hypertension. A hypothesis now under test is whether in both disorders, under prevailing conditions of ongoing stress, PNMT induced in sympathetic nerves acts as a DNA methylase, causing the norepinephrine transporter (NET) gene silencing that is present in both conditions. PNMT can have an intranuclear distribution, binding to DNA. We have demonstrated that the reduced neuronal noradrenaline reuptake present in both disorders does have an epigenetic mechanism, with demonstrable reduction in the abundance of the transporter protein, the NET gene silencing being associated with DNA binding by the methylation-related inhibitory transcription factor MeCP2. PMID- 19120128 TI - Vasopressin does not mediate hypersensitivity of the hypothalamic pituitary adrenal axis during chronic stress. AB - The hypothesis that vasopressin (VP) becomes the main mediator of pituitary corticotroph responsiveness during chronic hypothalamic pituitary adrenal (HPA) axis activation was tested by examining the effect of pharmacologic VP receptor blockade on the adrenocorticotropic hormone (ACTH) and corticosterone responses of 14-day repeatedly restrained rats. In spite of the increased vasopressinergic activity, repeatedly restrained rats showed lower ACTH and corticosterone responses to 10 min white noise compared with handled controls. These responses were unchanged by injection of the nonpeptide-selective V1b receptor antagonist SSR149415 i.v., 1 h before noise application. In contrast to noise stress, plasma ACTH responses to i.p. hypertonic saline injection were enhanced in the repeatedly restrained rats compared with handled controls, but responses were also unaffected by SSR149415 administered orally, daily 1 h before restraint. Since SSR149415 effectiveness was low, we used minipump infusion of the peptide V1 receptor antagonist, dGly[Phaa1,D-tyr(et), Lys, Arg]VP (V1-Ant) for 14 days, which effectively blocked ACTH responses to exogenous VP. Chronic V1-Ant infusion reduced plasma ACTH responses to i.p. hypertonic saline in handled controls but not in repeatedly restrained rats. These data suggest that the increased vasopressinergic activity characteristic of chronic stress plays roles other than mediating the hypersensitivity of the HPA axis to a novel stress. PMID- 19120129 TI - Peripherally administered angiotensin II AT1 receptor antagonists are anti-stress compounds in vivo. AB - Angiotensin II AT(1) receptor blockers (ARBs) are commonly used in the clinical treatment of hypertension. Subcutaneous or oral administration of the ARB candesartan inhibits brain as well as peripheral AT(1) receptors, indicating transport across the blood-brain barrier. Pretreatment with candesartan profoundly modifies the response to stress. The ARB prevents the peripheral and central sympathetic activation characteristic of isolation stress and abolishes the activation of the hypothalamic-pituitary-adrenal axis during isolation. In addition, candesartan prevents the isolation-induced decrease in cortical corticotropin-releasing factor 1 and benzodiazepine receptors induced by isolation. When administered before cold-restraint stress, candesartan totally prevents the production of gastric ulcerations. This preventive effect of candesartan is the consequence of profound anti-inflammatory effects, reduction of sympathetic stimulation, and preservation of blood flow to the gastric mucosa. The ARB does not reduce the hypothalamic-pituitary-adrenal axis stimulation during cold restraint. Preservation of the effects of endogenous glucocorticoids is essential for protection of the gastric mucosa during cold restraint. Administration of the ARB to nonstressed rats decreases anxiety in the elevated plus-maze. Our results demonstrate that Angiotensin II, through AT(1) receptor stimulation, is a major stress hormone, and that ARBs, in addition to their antihypertensive effects, may be considered for the treatment of stress-related disorders. PMID- 19120130 TI - Regulation of adrenoceptor and muscarinic receptor gene expression after single and repeated stress. AB - Although stress is tightly connected with elevated levels of catecholamines, stress effects on target structures of catecholamine action--adrenoceptors (ARs)- has not been deeply studied yet. Similarly, very little is known about changes of muscarinic receptors (MRs) during stress. We determined changes in these receptors in the individual parts of the heart (right atria and ventricles) of animals (rats and mice) exposed to a single and repeated immobilization stress. Changes of tissue catecholamines, beta(2)-AR gene expression, protein levels, and binding sites were determined in rat right ventricles, and changes in beta(1)-, beta(2)-, and beta(3)-AR gene expression were followed in murine right atria. Tissue catecholamines were elevated, while beta(2)-AR mRNA levels and beta(2)-AR proteins and binding were decreased, in rat right ventricles. In murine right atria, beta(1)- and beta(2)-AR gene expression was elevated, while beta(3)-AR mRNA levels and M(2)-MR were reduced. Taken together, our data show that interaction of AR and MR is important for the organism coping with stress and that different heart regions reveal distinct reactions to stress. PMID- 19120131 TI - Adrenoceptors and adaptive mechanisms in the heart during stress. AB - Several cardiovascular disorders have been related to alterations in beta adrenoceptor (beta-AR) signaling at or beyond the receptor level. During the stress reaction, the sympathetic-adrenal medullary system and the hypothalamus pituitary-adrenal cortex axis are activated, causing beta-AR overstimulation and remodeling of the beta(1)/beta(2)/beta(3)-AR ratio in cardiomyocytes. In a model of foot-shock stress, we described decreased beta(1)-AR signaling occurring simultaneously with increased beta(2)-AR signaling, whereas the response to the nonconventional agonist, CGP12177, was not altered. These alterations may play an adaptive role to the increased sympathetic drive to the heart, protecting the cardiac tissue from the cardiotoxic effects mediated by beta(1)-ARs overstimulation without altering cardiac output, since this would be sustained by the beta(2)-AR, which would also protect myocytes from apoptosis. Moreover, the selective enhancement of the beta(2)-AR population might help to diminish the risk of overstimulation since this adrenoceptor subtype couples to both, stimulatory G (Gs) and inhibitory G (Gi) proteins. On the other hand, in the model of neurogenic hypertension, the decrease in beta(1)-AR-mediated response is not followed by increase in the beta(2)-AR-mediated response. However, the response to CGP12177, which was desensitized 48 h after the surgery, was normalized 7 days after that, when beta(1)-AR were downregulated. Therefore, both experimental models provided evidence that the classical isoform of beta(1)-AR and the recently described low-affinity isoform of beta(1)-AR show independent behavior and provide the heart with adaptive mechanisms to increased sympathetic stimulation during stress. PMID- 19120132 TI - GABA regulates the rat hypothalamic-pituitary-adrenocortical axis via different GABA-A receptor alpha-subtypes. AB - The control of the corticotropin-releasing hormone (CRH) neurons in the hypothalamic paraventricular nucleus (PVN) is balanced by excitatory and inhibitory inputs. The GABA-A receptor, which is a major target for the inhibitory control, is composed of five subunits. The presence of an alpha(1)-, alpha(2)-, alpha(3)-, or alpha(5)-subunit in the GABA-A receptor protein complex is necessary for benzodiazepines to exert their potentiating effect on the receptor. We postulate that the effect of nonselective benzodiazepines on the hypothalamo-pituitary adrenocortical (HPA) axis is critically dependent on the composition of the GABA-A receptor subunits through which they act. We show here that positive modulators of alpha(1)-subtype containing GABA-A receptors with zolpidem (10 mg/kg) increase HPA activity in terms of increase in plasma corticosterone and induction of Fos in the PVN, whereas activation of non alpha(1)-subtype GABA-A receptors using L-818,417 (10 mg/kg) likely inhibits the activity. We also show that the alpha(2)-subunit gene is highly expressed in the PVN, but its expression is not affected by chronic mild stress. These results show that the stimulatory effect on HPA activity after positive modulation of GABA-A receptors composed of alpha(1)-subunit(s) affects a selective afferent system than the PVN, which is distinct from another afferent system(s) activated by non alpha(1)-containing GABA-A receptors. PMID- 19120133 TI - Gene expression of adrenoceptors in the hearts of cold-acclimated rats exposed to a novel stressor. AB - Changes in the heart rate and force of contraction are regulated by catecholamines via adrenoceptors (AR). In this work, we measured gene expression of AR in left heart atria and ventricles in rats exposed to cold stress and in cold-acclimated rats exposed to a novel stressor (immobilization). We found a significant increase in beta(3)-AR in left ventricle of rats exposed to acute (1 day) and long-term (28 days) cold, but no changes in beta(1)- and beta(2)-AR mRNA levels. However, single immobilization significantly decreased beta(2)-AR mRNA levels both in left atria and ventricles compared to acute cold stress. Application of a novel stressor (immobilization) to previously cold-acclimated animals did not show decrease of beta(2)-AR mRNA levels as seen in intact animals. Moreover, beta(1)- and beta(2)-AR did not show any significant changes. Surprisingly, the most prominent changes in the heart were detected for alpha(1B) AR gene expression. We found decreased levels of alpha(1B)-AR mRNA in the heart of rats exposed to cold and immobilization. We also found that exposure of cold acclimated rats to immobilization is responsible for additional decrease of alpha(1B)-AR mRNA levels in heart. It seems that while beta-AR undergoes adaptation, alpha(1B)-AR is probably prepared to modulate heart functions. Proposed mechanism of beta-AR adaptation needs to be elucidated. Thus, we have shown that gene expression of different AR subtypes in the heart is regulated differently by various stressors. A protective role of beta(2)-, beta(3)-AR, and alpha(1B)-AR in the process of heart adaptation to chronic stress exposure is proposed. PMID- 19120134 TI - Heart adrenoceptor gene expression and binding sites in the human failing heart. AB - Adrenergic regulation of the heart function is well documented by many studies. Catecholamines act through alpha(1)-, beta(1)-, beta(2)-, and beta(3) adrenoceptors (ARs) in the heart. There are many findings about the changes of beta(1)- and beta(2)-AR in heart failure (HF). On the other hand, the role of other AR subtypes is not clear yet. We focused on determining how HF could affect gene expression and specific ligand binding to alpha(1A)-, alpha(1B)-, alpha(1D) , beta(1)-, beta(2)-, and beta(3)-AR. Hearts from 11 patients with HF subjected to transplantation were investigated. As a control, corresponding parts from hearts not suitable for transplantation were used. We have found significantly higher mRNA levels of alpha(1A)-, alpha(1B)-,beta(1)-, and beta(2)-AR in the left ventricle of failing hearts compared to the levels in controls. beta(3)-AR mRNA levels in the left ventricle of failing hearts were not changed. No changes in mRNA levels of all receptors studied in other cardiac areas were found. On the other hand, binding studies showed a substantial decrease in left ventricles of failing hearts in all alpha(1)-AR subtypes and in beta(1)- and beta(2)-AR. However, the binding to beta(3)-AR was not changed. Our results suggest that alpha(1)-AR changes might be part of a compensatory mechanism, by which the heart suffering from the HF tries to secure its function, and it could be hypothesized that ineffective beta(3)-AR regulation might be involved in development of HF. According to our knowledge, this is the first report about the beta(3)-AR binding in HF. PMID- 19120135 TI - Neonatal programming of rat behavior by downregulation of alpha2A-adrenoreceptor gene expression in the brain. AB - Short-term knockdown of alpha2A-adrenergic receptor gene expression in the rat brain by siRNA or antisense oligodeoxynucleotide during the first days of life induced acute and long-lasting neurochemical and behavioral alterations. The acute effects in the neonatal rats were consistent with the known functions of the alpha2A-adrenergic receptors in the mature animals. The long-lasting alterations suggested involvement of receptor-specific gene expression during the critical period of brain development in early-life programming of anxiety-related behavior. PMID- 19120136 TI - Corticotropin-releasing hormone receptor subtypes in the rat anterior pituitary after two types of restraint stress. AB - The stress response in anterior pituitary (AP) is mediated by corticotropin releasing hormone (CRH) acting through CRH-R1 and -R2, however, the function of CRH-R2 in AP is still not fully elucidated. We used 1-h long restraint (IMO) as well as restraint combined with water immersion (IMO+C). Using real-time PCR we quantified mRNA expression of CRH-R1, CRH-R2alpha-soluble and -insoluble, and cAMP response element binding (CREB), with reference gene GAPDH. In control AP, CRH-R1 mRNA was up to 20-fold higher than levels of CRH-R2alpha-soluble or CRH R2alpha-insoluble mRNA. IMO reduced CRH-R1 mRNA to 47% and 63% of control levels 1 and 2 h after the onset of stressor, respectively, while IMO+C did not produce significant changes. Our data demonstrated that these stressors did not change CRH-R2alpha mRNA, unlike the very significant response of CRH-R1 to IMO. PMID- 19120137 TI - Hypoxia differently modulates gene expression of inositol 1,4,5-trisphosphate receptors in mouse kidney and HEK 293 cell line. AB - Hypoxia is a state of insufficient oxygen supply of the tissue or cell. Kidney tissue is highly sensitive to oxygen deprivation and easily develops renal ischemic injury. Calcium transporters very sensitively react to oxygen deficiency. We investigated whether hypoxia affects the gene expression of intracellular calcium transporters in the intact kidney, and we compared the response to that of HEK 293 cells. Our results showed that, while in mouse kidney tissue hypoxia elevates mRNA for inositol 1,4,5-trisphosphate receptors (IP3R) type 1 (IP3R1) and ryanodine receptors (RyR) type 2 (RyR2), in culture of HEK 293 cells the gene expression of all IP3Rs decreased without affecting viability of the cells. RyR2 mRNA in HEK 293 cells was not significantly changed, but RyR1 gene expression was significantly increased by hypoxia. The different response of kidney tissue and HEK 293 cells to hypoxia could be due to unequal differentiation state of the cells in intact tissue and cultured embryonic cell line. The physiological relevance of this observation remains to be determined. PMID- 19120138 TI - Reduced hypothalamo-pituitary-adrenal axis stress responses in late pregnancy: central opioid inhibition and noradrenergic mechanisms. AB - In late pregnancy, the hypothalamo-pituitary-adrenal (HPA) axis is less responsive to a range of psychological and physical stressors as a result of reduced central drive to the corticotropin-releasing hormone (CRH) neurons in the paraventricular nucleus (PVN). Most stressors activate the brain stem noradrenergic system, which innervates the majority of networks involved in regulating stress responses, including the PVN. Forced swimming, systemic interleukin-1beta (IL-1beta), and cholecystokinin (CCK) all activate brain stem noradrenergic cell groups, stimulate noradrenaline release in the PVN, and activate the HPA axis in nonpregnant rats. However, in late pregnancy we have shown that forced swimming and IL-1beta fail to evoke noradrenaline release in the PVN and hence HPA axis responses are suppressed. HPA axis responses to IL 1beta and CCK can be reinstated in pregnant rats by systemic administration of the opioid receptor antagonist naloxone, and when infused directly into the PVN, naloxone restores noradrenaline release in the PVN following IL-1beta treatment. Adrenaline release into the blood following stress is also attenuated in late pregnancy, despite increased adrenomedullary expression of tyrosine hydroxylase mRNA at this time. This review describes the mechanisms underlying attenuated HPA axis stress responses in pregnancy, focusing on the role of endogenous opioids and the central noradrenergic system. PMID- 19120139 TI - Gender-specific regulation of the hypothalamo-pituitary-adrenal axis and the role of vasopressin during the neonatal period. AB - Studies in arginine vasopressin (AVP)-deficient Brattleboro rats suggest that AVP is the predominant secretagogue during the perinatal period. Here we tested the hypothesis that congenital lack of vasopressin differentially modifies the stress reactivity of male and female rat pups. Vasopressin-producing (heterozygous, AVP+) and AVP-deficient (AVP-) Brattleboro rat pups of both genders were used. In 10-day-old pups, 24-h maternal separation and single, as well as repeated, ether inhalation induced remarkable adrenocorticotropin (ACTH) elevation only in AVP+ pups, supporting the role of vasopressin in hypothalamo-pituitary-adrenal (HPA) axis regulation. Surprisingly, the corticosterone elevations were even more pronounced in AVP- pups, suggesting the possibility of an ACTH-independent corticosterone-secretion regulation. In the case of maternal separation, both the plasma ACTH and corticosterone levels were higher in females than in males, while in case of ether inhalation only the ACTH levels were higher in females. Gender did not influence the stress reactivity or the effect of the genotype. We conclude that the gender of the pups did not profoundly influence HPA axis activity (the mechanism seems to be the same), but in contrast to the general view, we suggest that the females have a more active HPA axis than the males already during the neonatal period. However, the resting corticosterone elevation well known in adult females- is missing. PMID- 19120140 TI - Sex-specific prenatal programming: a risk for fibromyalgia? AB - Women are four to eight times more likely to be affected by fibromyalgia syndrome (FMS). A lack of cortisol, potentially due to an adrenocortical deficit is postulated in FMS. The cause of such adrenal insufficiency is unknown. It could be assumed that stress exposure during critical periods contributes to vulnerability for FMS. These critical periods might include prenatal periods in which adversities may lead to an impaired development of the adrenal cortex, especially in females. More than 50% of FMS patients report major life events before the onset of the disease. Possibly due to adrenal insufficiency they may not be able to dampen their stress response by secreting sufficient glucocorticoids. Thus, stress mediators, such as catecholamines and pro inflammatory cytokines, may be disinhibited and affect brain function. This might result in an enhanced responsiveness to external and internal pain- and fatigue eliciting stimuli. In a study with female FMS patients (N= 93) those patients with a shorter gestational length (<38 weeks) showed a lower cortisol awakening response (CAR) than FMS subjects with a gestational length >38 weeks (F((3,31))= 2.94, P= 0.038). Additionally, more than 70% reported severe psychological stress alone or in combination with other factors at disease onset. PMID- 19120141 TI - Response of the adrenomedullary system to early postnatal stress in the Brattleboro rat. AB - The sympathetic-adrenomedullary system and the pituitary-adrenocortical axis are linked to each other by chemical signals transferring information between both endocrine systems. Here we addressed the question of whether the neuropeptide arginine vasopressin (AVP) is involved in this type of information transfer during early postnatal development. The impact of congenital absence of AVP on the endocrine stress response was investigated using the AVP-deficient Brattleboro rat. Under resting conditions, we failed measure a significant difference in plasma norepinephrine levels between 10-day-old AVP-deficient homozygous juveniles versus heterozygous AVP-producing littermates. Interestingly, repeated ether exposure resulted in a reduction of plasma epinephrine levels in both genotypes. In the adrenal, we detected increased levels of the epinephrine-synthesizing enzyme phenylethanolamine-N methyltransferase (PNMT) after ether inhalation in vasopressin-deficient pups only. These data provide insight into the development of the regulation of stress related epinephrine secretion during ontogenesis. Furthermore, our results imply that the congenital absence of AVP affects the synthesis of PNMT in response to defined stressor exposure. PMID- 19120142 TI - Pheochromocytoma: an endocrine stress mimicking disorder. AB - A pheochromocytoma is an endocrine tumor that can uniquely mimic numerous stress associated disorders, with variations in clinical manifestations resulting from different patterns of catecholamine secretion and actions of released catecholamines on physiological systems. PMID- 19120143 TI - Unexplained symptomatic paroxysmal hypertension in pseudopheochromocytoma: a stress response disorder? AB - Among overall numbers of patients tested for pheochromocytoma, less than 2% harbor the tumor. Among the rest, there is often no satisfactory explanation for the signs and symptoms leading to suspicion of pheochromocytoma. This group includes patients with severe symptomatic paroxysmal hypertension, often referred to as pseudopheochromocytoma, a condition that can be debilitating for patients and perplexing for clinicians. Similar to patients with the real tumor, patients with pseudopheochromocytoma can be misdiagnosed with panic disorder. However, pseudopheochromocytoma is characterized by an absence of panic or emotional distress preceding the onset of hypertension and symptoms of catecholamine excess. Because the clinical manifestations of pseudopheochromocytoma are similar, if not identical, to those due to excess circulating catecholamines in patients with the tumor, the most attractive explanation for the disorder is that it involves altered function of the autonomic nervous system. In line with this hypothesis, recent findings suggest that enhanced adrenal release of epinephrine and exaggerated cardiovascular responsiveness to catecholamines both contribute to the paroxysmal hypertension and symptoms of catecholamine excess in pseudopheochromocytoma. From this pattern, one would predict that therapeutic interventions that inhibit adrenal secretion of epinephrine or block adrenoceptor mediated responses to catecholamines might provide a logical approach to therapy. PMID- 19120144 TI - Catecholamines and estrogen are involved in the pathogenesis of emotional stress induced acute heart attack. AB - Emotional stress triggers takotsubo cardiomyopathy in postmenopausal women. Clinical analysis of autonomic nervous function has revealed a transient increase of sympathetic nervous activity and decrease of vagal nervous activity. Immobilization (IMO) stress of rats can reproduce the electrocardiographic and left ventriculographic changes that occur in takotsubo cardiomyopathy, both of which are prevented by combined blockade of alpha- and beta-adrenoceptors. Estrogen supplementation partially attenuated these cardiac changes. It also attenuated the IMO-induced increase of c-Fos immunoreactivity, or c-fos mRNA expression in the lateral septum, medial amygdaloid nucleus, paraventricular hypothalamic nucleus, dorsomedial hypothalamic nucleus, laterodorsal tegmental nucleus, and locus ceruleus; these regions contain central sympathetic neurons and neurons with immunoreactive estrogen receptors. It also downregulated c-fos mRNA expression in the adrenal gland and the heart, suggesting an increase of estrogen attenuated the stress-induced hypothalamo-sympathoadrenal outflow from the central nervous system to the target organs. Estrogen treatment also upregulated the levels of cardioprotective substances, such as atrial natriuretic peptide and heat shock protein 70, in the heart. These data suggest that reduction of estrogen levels following menopause might be involved in the primary cause of takotsubo cardiomyopathy both by indirect action on the nervous system and by direct action on the heart. PMID- 19120145 TI - Attenuated pre-ejection period response to tyramine in patients with cardiac sympathetic denervation. AB - Stress is a well-known factor affecting cardiac contractility through the cardiac sympathetic nerves. A positive inotropic effect of the cardiac sympathetic nerves on the myocardium is reflected by pre-ejection period (PEP) shortening. Patients with Parkinson disease (PD) and neurogenic orthostatic hypotension (NOH) (PD + NOH) or with pure autonomic failure (PAF) have markedly decreased myocardial 6 [(18)F]Fluorodopamine-derived radioactivity, reflecting cardiac sympathetic denervation. The functional effects of the cardiac sympathetic denervation have been unknown. We measured PEP and heart rate-corrected PEP (PEPI) responses to i.v. tyramine (1 mg/min) in 13 patients (9 PD + NOH and 4 PAF) with low 6 [(18)F]Fluorodopamine-derived radioactivity and in subjects with normal radioactivity (15 multiple system atrophy with NOS patients (MSA + NOS). Baseline PEP and PEPI did not differ between the groups. By 10 min after initiation of tyramine infusion, PEP and PEPI were significantly lower (P < 0.01) in MSA + NOS, compared to base line, whereas PEP and PEPI remained unchanged in the PD + NOH/PAF group. The PEP and PEPI decrease was larger in the MSA + NOS group than in the PD + NOH/PAF group (P < 0.05). One of the functional consequences of cardiac sympathetic denervation is failure to increase contractility in response to stimuli that depend on endogenous norepinephrine release. PMID- 19120146 TI - The role of norepinephrine and insulin resistance in an early stage of hypertension. AB - The interrelationship between activity of sympathetic nervous system and metabolic risk factors in youth with hypertension (HT) has been poorly studied. The aim of our present study was to assess the interrelationship between metabolic risk factors, such as insulin resistance, concentration of plasminogen activator inhibitor (PAI)-1, and catecholamines in an early stage of HT onset. An oral glucose tolerance test was performed in 17 young males with early-diagnosed nontreated HT grade 1 and 16 gender-, age-, and BMI-matched normotensive controls. Concentrations of glucose, insulin, epinephrine, norepinephrine, PAI-1, and plasma renin activity (PRA) were determined in venous plasma. Insulin sensitivity indices (ISIs) proposed by Cederholm, Matsuda, and Gutt were calculated. HT had higher baseline levels of norepinephrine, insulin (P= 0.02), and PAI-1 (P= 0.04). ISIs were lower in HT subjects (P < 0.001). Baseline concentrations of epinephrine were negatively associated with HDL cholesterol (r= 0.415, P= 0.02), ISI Matsuda (r=-0.361, P= 0.04), ISI Cederholm (r=-0.354, P= 0.04), and ISI Gutt (r=-0.429, P= 0.01), and positively with PRA (r= 0.609, P < 0.0001). Positive association was found between baseline concentrations of norepinephrine and PAI-1 (r= 0.418, P= 0.02). The sympathetic overactivity, which occurs in the early stage of HT may contribute to reduced insulin sensitivity even in young patients and intensify the undesirable development of metabolic cardiovascular risk factors and progress of the disease. PMID- 19120147 TI - Endocrine factors in stress and psychiatric disorders: focus on anxiety and salivary steroids. AB - Glucocorticoids and other steroids produced in the adrenal cortex are altered in chronic stress situations associated with enhanced anxiety. A useful tool to evaluate changes in adrenal steroids during stress and anxiety under both laboratory and real-life stress situations is determination of steroids in saliva. The main advantages of this technique are its noninvasiveness and its measurement of biologically active free hormone levels. Salivary cortisol is a valuable indicator of the activity of the hypothalamic-pituitary-adrenocortical axis, which is known to be altered in psychiatric disorders. Measurements of salivary cortisol helped to reveal changes that would otherwise remained undetected, such as increase in cortisol release in spontaneously occurring panic attacks. By selecting only the subjects with high and low trait anxiety, we have brought evidence confirmed by others that high trait anxiety may be associated with an inability to respond with adequate cortisol release during stress. Papers on the relationship between salivary dehydroepiandrosterone and trait anxiety or anxiety disorders are rare, and this stress hormone deserves more attention. Almost nothing is known on aldosterone and anxiety. We have modified the methodology of aldosterone radioimmunoassay by concentrating the saliva and validated it biologically by demonstrating daily variation and gender differences. We have provided the first data on the relationship between aldosterone and trait anxiety. Obtained results show a significant negative correlation between morning salivary aldosterone concentrations and trait anxiety scores in women (luteal phase), but not in men. A more proper elucidation of the association between aldosterone and anxiety seems to be an important target of further research. PMID- 19120148 TI - Salivary cortisol levels in Brazilian citizens of distinct socioeconomic and cultural levels. AB - We have analyzed the perceived stress index, the basal salivary cortisol levels, and the awakening cortisol response (ACR) in 86 volunteers of low (LSES) and high socioeconomic status (HSES). The LSES presented higher perceived stress index and basal salivary cortisol levels, nonaltered ACR, or cortisol diurnal rhythm. We have concluded that the LSES is associated with high perceived stress index and salivary cortisol levels, which could impact negatively in health, and that it is related to the daily life stress experienced by individuals in the LSES group. Because the LSES corresponds to about 30% of the total Brazilian population, this conclusion might have a great impact on public health policies and costs. PMID- 19120149 TI - Corticosterone regulates pERK1/2 map kinase in a chronic depression model. AB - Neurotransmitter- or neurotrophin-regulated intracellular signaling in the hippocampus is hypothesized to contribute to depression and antidepressant (ADT) efficacy. Extracellular signal-regulated kinase 1/2 (ERK1/2) is downstream of several receptor types and regulates transcriptional activity of many targets; ERK1/2 may thereby influence mood and affect. Using a novel, ADT-sensitive depression model in mice, we show that prior corticosterone exposure decreases motivated behavior, sucrose consumption, and pERK1/2 in the dentate gyrus, but not in CA1/CA3. Notably, prefrontal cortical targets were also regulated. Our data suggest ADTs restore hippocampal pERK1/2 after stress-related insult, and potentially reveal a novel role for prefrontal neurotrophins in depressive-like symptomology. PMID- 19120150 TI - Fast action of neuroactive steroids on plasma membrane calcium pump in PC12 cells. AB - Calcium ions are essential to proper neurotransmission. Impairment in cytosolic Ca(2+) concentration and Ca(2+) signaling disturbs neuronal activity, leading to pathological consequences. In cells, a high-affinity plasma membrane calcium pump (PMCA) keeps free Ca(2+) in the nanomolar range. Among four genes encoding the enzyme, PMCA2 and 3 are primary in excitable cells. To elucidate the relationship between PMCAs' composition and susceptibility for neurosteroid regulation, we obtained PC12 cells with suppressed neuron-specific isoforms and analyzed the effect of selected steroids on Ca(2+) uptake. Our results indicate that hormones affected Ca(2+) transport activity and that this effect depended on both PMCA isoform composition and steroid structure. PMID- 19120151 TI - Neuroendocrine activation during combined mental and physical stress in women depends on trait anxiety and the phase of the menstrual cycle. AB - The aim of this study was to investigate the influence of trait anxiety and menstrual cycle phase on neuroendocrine activation during combined mental and physical stress procedure in 40 healthy female subjects. Women at the upper (anxious) and lower (nonanxious) limits of the normal range of a trait anxiety scale were exposed to the stress procedure consisting of a mental component (Stroop test) and handgrip exercise. Salivary cortisol levels, cardiovascular parameters, and cognitive performance in the Stroop test were evaluated. Stress induced cortisol levels and the rise in systolic blood pressure were affected by both trait anxiety and menstrual cycle phase. The stress model used induced a significant cortisol elevation only in anxious women in the follicular phase. This group of women also exhibited greater increases in systolic blood pressure in response to handgrip exercise as compared to anxious ones in the luteal phase and to nonanxious women in either phase. In nonanxious women, stress-induced cortisol levels positively correlated with cognitive performance. In contrast, a negative correlation trend was observed in anxious subjects. Thus, in subjects with low but not high trait anxiety, enhanced cortisol concentrations seem to be associated with better cognitive performance. The results suggest that women with high trait anxiety exhibit greater cardiovascular and hormonal sensitivity to stress stimuli during the follicular phase. PMID- 19120152 TI - Drive for leanness, anorexia nervosa, and overactivity: the missing link. AB - Anorexia nervosa (AN) is a psychiatric condition characterized by deliberate reduction of body weight. Some patients with AN exhibit overactivity that can worsen their state of malnutrition. Employing an evolutionary psychiatry line of inquiry, we propose that rigidity of thinking and overactivity are behavioral phenotypic changes in AN patients that are normal to tree-dwelling mammals, such as monkeys. Such behavior can lead to good functioning as ballet dancers and athletes but lead to certain disadvantages in other areas of modern life. The overactivity in AN, though under conscious control may be neurobehavioral and driven subconsciously by disordered cerebral neuropsychopharmacology. PMID- 19120153 TI - Rotenone damages striatal organotypic slice culture. AB - Epidemiological studies suggest the involvement of pesticides in the etiology of Parkinson's disease. Exposure to rotenone results in degeneration of the nigrostriatal pathway through inhibition of complex I. Organotypic striatal slice cultures were prepared from brains of adult mice and treated with rotenone (0.01, 0.05, 0.1, and 1 mM) for 48 h. Lactate dehydrogenase activity was elevated by 167% at 1 mM of rotenone. Using fluorescent indicators, membrane damage was up to 130% as measured by propidium iodide fluorescence, and superoxide (DHE) and nitric oxide (DAF-FM) formation were increased by 195% and 774% at 1 mM of rotenone, respectively, compared to controls. The study concludes that formation of radicals mediated striatal degeneration by rotenone. PMID- 19120154 TI - Differential effects of the new glucocorticoid receptor antagonist ORG 34517 and RU486 (mifepristone) on glucocorticoid receptor nuclear translocation in the AtT20 cell line. AB - Glucocorticoid agonists bind to cytoplasmic glucocorticoid receptors (GRs) and subsequently translocate as an agonist-GR complex into the nucleus. In the nucleus the complex regulates the transcription of target genes. A number of GR antagonists (RU486, progesterone, RU40555) have also been shown to induce receptor translocation. These compounds should be regarded as partial agonists. For the nonselective progesterone receptor antagonists, RTI3021-012 and RTI3021 022, it was shown that GR antagonism is possible without the induction of GR translocation. In the present studies, the new GR antagonist, ORG 34517, was investigated for its potential to induce GR translocation and to antagonize corticosterone-induced GR translocation in the AtT20 (mouse pituitary) cell line. ORG 34517 was compared to RU486. In contrast to RU486, ORG 34517 (at doses up to 3 x 10(-7) M) did not induce GR translocation, but was able to block corticosterone (3 x 10(-8) M) induced GR translocation. ORG 34517 can be regarded as a true competitive GR antagonist without partial agonistic activities. PMID- 19120155 TI - Ethanol and stress activate catecholamine synthesis in the adrenal: effects on bone. AB - Ethanol consumption and mental stress activate the sympathetic nervous system, which can adversely affect bone. We compared six groups of 10 young adult rats, three with and three without 2 h daily restraint stress. Two groups consumed food and water ad libitum, two received food and 6% (w/v) ethanol as drinking water, and two received the amount of food consumed by ethanol rats the previous day plus water ad libitum (pairfed). After 6 weeks, rats were killed. Plasma, femurs, lumbar vertebrae, and adrenals were harvested. Femoral dimensions were measured and biomechanical properties were tested by three-point bending. Plasma osteocalcin, vertebral osteocalcin mRNA levels, and adrenomedullary tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine N methyl transferase (PNMT) mRNA levels were quantified. Daily restraint decreased weight gain and femoral length compared to dietary controls, and appeared to partially preserve bone strength, especially in calorie-restricted pairfed rats. Femoral strength was significantly affected by treatment in that bones of pairfed controls were weakest, ethanol drinkers were intermediate, and ad libitum restrained were strongest. Femoral yield load, displacement, and work at yield load were negatively correlated with TH and DBH mRNA levels, but not PNMT, suggesting a negative influence of norepinephrine. Plasma osteocalcin and dry weight of lumbar 3-5 vertebrae were unaffected; however, osteocalcin mRNA in second lumbar vertebrae was positively correlated with TH, DBH, and PNMT levels. Ethanol consumption at this level had little effect on femur morphology or strength. In contrast, the data suggested possible stimulation rather than inhibition of vertebral bone formation. PMID- 19120156 TI - Adrenomedullary response to hypoglycemia in first-degree relatives of patients with rheumatoid arthritis. AB - Our recent studies showed blunted adrenomedullary responses to insulin-induced hypoglycemia in premenopausal females with rheumatoid arthritis (RA) and systemic sclerosis, suggesting dysregulation of the adrenomedullary hormonal system (AMHS). Since no relationship has been found between degree of AMHS dysfunction and clinical or inflammatory parameters in those patients, we hypothesize the presence of an inherited perturbation of the AMHS. To test this hypothesis, we evaluated adrenomedullary responses to insulin-induced hypoglycemia (0.1 IU/kg) in premenopausal female subjects: 17 glucocorticoid-naive RA patients, 15 healthy first-degree family members (FDR), and 18 age- and body mass index-matched healthy controls. Our results demonstrate that when compared to controls, RA patients had lower baseline epinephrine levels (P= 0.01) and lower area under response curve (AUC) levels of norepinephrine (P < 0.001) and epinephrine (P < 0.003). In contrast, FDR had lower (P= 0.001) AUC levels of norepinephrine compared to controls and higher (P= 0.033) AUC levels of epinephrine compared to RA patients. There were no significant differences in epinephrine response between FDR and controls. Although we found lower norepinephrine responses to hypoglycemia in FDR of RA patients, adrenomedullary responses to hypoglycemia does not appear to be altered to the degree found in RA patients. PMID- 19120157 TI - Sympathetic nervous system response to orthostatic stress in female patients with rheumatoid arthritis. AB - Sympathoneural and adrenomedullary impairments have been suggested in patients with rheumatoid arthritis (RA). In the present study, sympathoneural and adrenomedullary responses to orthostasis were evaluated in eight female RA patients and in eight matched healthy controls. The testing consisted of sequence of stabilization period in supine position, legs-up position, orthostasis, and supine position. In each body position, blood samples were drawn and ECG was recorded. Plasma levels of epinephrine (EPI), norepinephrine (NE) and neuropeptide Y (NPY) were measured and sympathoneural activity was evaluated by analysis of heart rate variability (HRV). Higher baseline NE levels were found in RA patients (P= 0.034), without any difference in response to orthostasis between the study groups. Levels of EPI tended to be lower in RA patients in base line (P= 0.053) and in response to orthostasis (P= 0.079). The RA and control groups did not differ in NPY levels or in HRV parameters considered to reflect sympathetic activity. A subtle tendency to decreased adrenomedullary reactivity but no evidence for abnormal sympathetic responses to orthostasis was found in RA females. PMID- 19120158 TI - Adrenocortical response to low-dose ACTH test in female patients with rheumatoid arthritis. AB - Alterations in adrenal steroid production have been suggested in females with rheumatoid arthritis (RA). The aim of the present study was to assess adrenocortical function in RA females. We examined 11 female RA patients (RA: age 30 +/- 2 years, BMI 21.0 +/- 0.7 kg/m(2)) and 10 matched healthy controls (C: age 31 +/- 1 years, BMI 21.6 +/- 0.6 kg/m(2)). Low-dose adrenocorticotropic hormone (ACTH) test (i.v. bolus of 1 microg synthetic ACTH) was performed at 10.00 h with blood sampling every 15 min for 90 min. Cortisol, 17-OH-progesterone (17OHP), androstenedione (ASD), and dehydroepiandrosterone (DHEA) were assayed in plasma. Baseline cortisol levels were higher in RA patients (RA: 385 +/- 38 versus C: 229 +/- 28 nmol/L, P= 0.007). In both study groups, ACTH administration increased all the four steroids measured (P < 0.001). Cortisol response to ACTH administration was diminished in RA patients when compared to controls (Delta(max): 284 +/- 24 in RA versus 424 +/- 31 nmol/L in C, P= 0.002). ACTH-induced maximal rise in plasma DHEA was significantly lower in RA patients when compared to controls (Delta(max): 2.59 +/- 0.68 in RA versus 5.57 +/- 1.25 ng/mL in C, P= 0.015). No significant between-groups differences were found in responses of ASD or 17OHP. The molar ratio of ASD:cortisol was significantly lower (P < 0.05) in RA patients at base line, but did not differ during ACTH test. After ACTH bolus, the cortisol:17OHP ratio decreased significantly in the RA group (P < 0.001), whereas there was no change in the control group. The present results show decreased secretion of cortisol and DHEA in RA patients in response to ACTH, suggesting a subtle HPA hypofunction at the adrenocortical level. PMID- 19120159 TI - Animal biodiversity and emerging diseases prediction and prevention. Introduction. PMID- 19120161 TI - Global factors driving emerging infectious diseases. AB - The extinction of species across the globe is accelerating directly or indirectly from human activities. Biological impoverishment, habitat fragmentation, climate change, increasing toxification, and the rapid global movement of people and other living organisms have worked synergistically to diminish ecosystem function. This has resulted in unprecedented levels of disease emergence driven by human-induced environmental degradation, which poses a threat to the survival and health of biodiversity. What is often overlooked in the discussion of the health consequences to humans is that critically endangered wildlife species are at grave risk of extinction by disease outbreaks. As habitat becomes more compressed and with migration routes cut off, gene pools of small species are stranded in isolated habitat fragments. Species now are vulnerable to encroachment, malnutrition, environmental pollutants, and epidemics from domestic animals and humans. Furthermore, the continuous degradation of ecosystems is leading to increased stress, immunosuppression, and greater susceptibility to disease. Disease can be catastrophic to a diminished stressed population, becoming in some instances the leading factor of local, regional, and global extinctions. The strategies of the new field of conservation medicine include long-term monitoring, health assessment, and interventions to protect species at risk. We particularly must minimize the threat of any potentially catastrophic disease outbreaks resulting from anthropogenic changes to the environment. Current and future diagnostic molecular techniques offer new opportunities to identify tools for the management and possible treatment of diseases in imperiled species. PMID- 19120162 TI - A resolution by the society for tropical veterinary medicine in support of "one health". AB - At its ninth biennial conference, the membership of the Society for Tropical Veterinary Medicine (STVM) adopted a resolution in support of "One Health," an initiative that promotes collaboration between veterinary and human medical professionals and allied health scientists. The resolution is published on the website of the STVM and has been shared with related veterinary medical and medical associations and societies. PMID- 19120163 TI - Community-based animal health care. AB - Livestock contribute to the livelihoods of around 70% of the world's poor. Heifer International provides high quality livestock, training, and related support to communities to assist with food security and to improve livelihoods. Heifer provides training and extension services so families can provide improved animal housing, management, breeding, nutrition, and veterinary health. Heifer conducts training programs for para-vets or Community Animal Health Workers (CAHWs) in areas where local professional veterinary care is not available. Beneficiaries get animal health and husbandry services through trained technicians who are selected from within the community and trained in improved local practices, using basic selection guidelines. The selected community men and women get hands-on training based on the principle of community-based animal health care, using participatory principles in the farmers' fields. Basic tasks of CAHWs in the Heifer-supported project are described. Heifer International is providing sustainable solutions to the well-being of the livestock where it works. PMID- 19120164 TI - The Caribbean animal health network: new tools for harmonization and reinforcement of animal disease surveillance. AB - The Caribbean Animal Health Network (CaribVET) is a collaboration of veterinary services, diagnostic laboratories, research institutes, universities, and regional/international organizations to improve animal health in the Caribbean. New tools were used by the network to develop regional animal health activities: (1) A steering committee, a coordination unit, and working groups on specific diseases or activities were established. The working group on avian influenza used a collaborative Web site to develop a regionally harmonized avian influenza surveillance protocol and performance indicators. (2) A specific network was implemented on West Nile virus (WNV) to describe the WNV status of the Caribbean countries, to perform a technology transfer of WNV diagnostics, and to establish a surveillance system. (3) The CaribVET Web site (http://www.caribvet.net) encompasses information on surveillance systems, diagnostic laboratories, conferences, bibliography, and diseases of major concern in the region. It is a participatory Web site allowing registered users to add or edit information, pages, or data. An online notification system of sanitary information was set up for Guadeloupe to improve knowledge on animal diseases and facilitate early alert. PMID- 19120165 TI - Infectious diseases and the illegal wildlife trade. AB - We provide a compilation of pathogens directly associated with illegally traded wildlife. We find that these pathogens span the gamut of taxonomic origins, affect most vertebrate taxa, and can have negative consequences for human and animal health and the global economy. Significantly, published health assessments of illegally traded wild animals are extremely scarce, making the results of this survey an underestimation of the true extent of the problem. Concerted action aimed at preventing further negative health effects is warranted. PMID- 19120166 TI - Tropical veterinary parasites at Harvard University's Museum of Comparative Zoology. AB - Tropical veterinary parasites have been maintained by the Museum of Comparative Zoology (MCZ) at Harvard University since the mid 1800s. Most of these are maintained by the Department of Invertebrate Zoology, but many vectors and intermediate hosts are maintained by the Departments of Entomology and Malacology. The largest collections are of avian and mammalian ticks (Acarina) that are important as both parasites and vectors. Nematodes are second in numbers, followed by cestodes, trematodes, and several minor helminth groups, crustacean parasites of fish, and protozoan parasites of various hosts. The MCZ directed or participated in several major expeditions to tropical areas around the globe in the early 1900s. Many of these expeditions focused on human parasites, but hundreds of veterinary and zoonotic parasites were also collected from these and numerous, smaller, tropical expeditions. Host sources include companion animals, livestock, laboratory species, domestic fowl, reptiles, amphibians, exotics/zoo animals, commercially important fishes, and other wildlife. Specimens are curated, either fixed whole in vials or mounted on slides as whole mounts or histopathological sections. The primary emphasis of MCZ's current work with tropical veterinary parasites is on voucher specimens from epidemiological, experimental, and clinical research. PMID- 19120167 TI - Use of plant-derived products to control arthropods of veterinary importance: a review. AB - The use of synthetic products in veterinary pest management is becoming increasingly problematic. Issues, including pest resistance, product withdrawal, undesirable environmental persistence, and high mammalian toxicity associated with synthetic pesticides, are driving research to identify new pest management approaches. One approach employs the repellent/toxic effects of plant-derived products (PDPs). Several pesticides based on PDPs are already available in some areas of pest management. This review highlights instances in which such products have been used with success against pests of domestic animals, livestock, apiculture, and poultry. PMID- 19120168 TI - Differential expression of the tick protective antigen subolesin in anaplasma marginale- and A. phagocytophilum-infected host cells. AB - Subolesin was recently shown in vaccine and RNA interference (RNAi) studies to protect against tick infestations and to affect tick feeding, reproduction, and development as well as infection of host cells by Anaplasma marginale and A. phagocytophilum. Recent experiments provided evidence that infection of both tick and vertebrate host cells with these two pathogens modified gene expression. We therefore hypothesized that infection of host cells with A. marginale and A. phagocytophilum affects expression of subolesin. Subolesin mRNA levels were determined by real-time reverse transcriptase (RT)-PCR in uninfected and A. marginale-infected Dermacentor variabilis guts and salivary glands and IDE8 cultured tick cells and in uninfected and A. phagocytophilum-infected Ixodes scapularis nymphs, ISE6-cultured tick cells, and the human cell line HL-60. In addition, the effect of subolesin on Anaplasma spp. infection/multiplication was characterized by RNAi in tick tissues and/or cultured tick and human cells. These experiments presented evidence of differential expression of subolesin in A. marginale- and A. phagocytophilum-infected cells. Subolesin was differentially expressed in A. marginale-infected ticks in a tissue-specific manner in which mRNA levels increased in response to A. marginale infection in tick salivary gland cells but not in the gut cells. Subolesin knockdown by RNAi reduced Anaplasma infection/multiplication only in cells in which infection increased subolesin expression, i.e., in A. marginale-infected D. variabilis salivary glands and IDE8 cells. The results reported herein further support the role of subolesin in Anaplasma-host interactions and suggest a putative role of subolesin in vaccines for the control of pathogen infection/multiplication in ticks. PMID- 19120169 TI - Immunological control of the poultry red mite. AB - In the current study whole poultry red mite antigens were extracted and birds were immunized subcutaneously with either antigen in adjuvant (antigen group) or PBS in adjuvant (control group). Immune responses of birds following immunization were investigated by ELISA and Western blotting, while vaccine efficacy was assessed by feeding of red mites on birds. Immunized birds showed a significant (P < 0.05) increase in IgY titers after immunization compared to controls, while immunoglobulin A (IgA) and IgM did not change significantly. However, the antigen group had a generally higher increase in all immunoglobulin titers compared to the controls. Western blotting identified a number of protein bands at different molecular weights, although these were not different between treatments. PCR analysis of whole mite protein identified bacterial DNA that might have confounded immunological data. In addition, there was a trend toward reduced survival rate of red mites feeding on antigen-immunized birds, but the difference was not statistically significant compared to controls. This study demonstrates the potential for somatic red mite antigens to stimulate an antibody-mediated immune response, although this response did not confer protection to birds. PMID- 19120170 TI - Defining the role of subolesin in tick cell culture by use of RNA interference. AB - Development of tick vaccines provides new opportunities for control of tick infestations and tick-borne diseases. Recently, the tick-protective protein, subolesin, was identified in a cell line derived from Ixodes scapularis by expression library immunization and a mouse model of tick infestations. While subolesin was conserved among ixodid tick species, the biological function of this gene is unknown. Subolesin expression in ticks was silenced by RNA interference (RNAi) to provide information on the gene's function, and silencing of subolesin profoundly impacted tick survival, feeding, and reproduction. In this research we used RNAi in the IDE8 tick cell line to further study the role of subolesin in development of cultured tick cells. The cells were incubated with subolesin double-stranded (ds)RNA and cell growth was monitored. Incorporation of dsRNA by tick cells was monitored with Cy3-labeled dsRNA. After 72 h cells were harvested for cell counts, morphology, and for confirmation of gene silencing by reverse transcriptase-PCR. While the expression of subolesin in treated cells was reduced 80 +/- 9% by RNAi as compared with mock-treated cells, cell growth did not appear to be affected over the 72-h period. This is the first report of the use of RNAi in tick cell culture. RNAi is a powerful tool for studying tick gene function and will likely contribute to our understanding of the role that tick genes play in cell development and infection with pathogens. PMID- 19120171 TI - Molecular cloning, sequence analysis, and immune recognition of Bm95 from Thai strains of Rhipicephalus (Boophilus) microplus. AB - The efficacy of existing vaccines against Thai strains of Rhipicephalus (Boophilus) microplus remains to be determined, and these vaccine antigens should be compared to homologues derived from local strains. The purpose of this study was to compare Bm95 from Thai R. microplus to the previously reported sequence from Argentinean ticks. mRNA was isolated from R. microplus midgut samples, and cDNA was amplified with Bm95-specific primers. The cDNA and protein sequences of Thai Bm95 were 94 and 92% identical, respectively, to Argentinean Bm95. Although the sequence was similar to Argentinean Bm95, there were 45-amino acid differences among the homologues. Amplicons encoding Bm95 were cloned into pPICZalphaA and expressed in Pichia pastoris strain KM71H. The recombinant plasmid Bm95 (rBm95) was reactive to bovine hyperimmune serum to Thai R. microplus midgut, indicating that the Thai rBm95 was immunogenic. Further work is needed to compare the efficacies of Thai and Argentinean rBm95 for protection of cattle against infestation with Thai R. microplus. PMID- 19120172 TI - Primary midgut, salivary gland, and ovary cultures from Boophilus microplus. AB - Primary cell cultures from different tick organs are a valuable tool for host parasite research in the study of the protozoan Babesia sp., which infects different organs of the tick. In this work we describe the generation of midgut, salivary gland, and ovary primary cell cultures from dissections of Boophilus microplus. Midguts, salivary glands, and ovaries were dissected from B. microplus ticks on different days after bovine infestation; different enzymatic disaggregating protocols were tested in the presence of proteolytic enzymes, such as trypsin and collagenase type I and II, for tissue disaggregation and primary cell culture generation. The dissected tick organs obtained 18-20 days after bovine infestation showed a major cellular differentiation and were easier to identify by cellular morphology. The enzymatic disaggregation results showed that each tissue required a different proteolytic enzyme for optimal disaggregation; collagenase type I produced the most complete disaggregation for ovaries but not for midgut or salivary glands. Collagenase type II was effective for salivary glands but performed poorly on ovaries and midgets, and typsin was effective for midguts only. The midgut and ovary primary cell cultures were maintained for 4 weeks in optimal conditions after the cells were no longer viable. The salivary gland cell cultures were viable for 8 months. PMID- 19120173 TI - Molecular analysis of calreticulin expressed in salivary glands of Rhipicephalus (Boophilus) microplus indigenous to Thailand. AB - The tropical cattle tick, Rhipicephalus (Boophilus) microplus, is an important ectoparasite of livestock in Thailand that causes economic losses due to the direct effects of tick feeding and by the pathogens they transmit. Intensive acaricide use has several drawbacks, which spurred efforts to develop anti-tick vaccines. Vaccines targeting concealed antigens localized in the tick midgut result in reduced tick fecundity, but molecules localized in the tick salivary glands, which could play a role in pathogen transmission, remain largely unexplored for R. microplus. Calreticulin (CRT) is a protein found in tick salivary glands and saliva, and CRT might facilitate tick feeding and pathogen transmission through anti-thrombotic and complement-inhibition activities. This then suggests that CRT should be evaluated as a vaccine candidate antigen to control cattle ticks in Thailand. The objective of this work was to clone, sequence, and analyze cDNA encoding CRT from salivary glands of R. microplus indigenous to Thailand. Nucleotide sequence analysis showed an open reading frame of 1233 bp. Comparison of the amino acid sequence showed 65-99% identities to other known CRTs from Oryctolagus cuniculus, Rattus norvegicus, Homo sapiens, Bos taurus, R. sanguineus, and R. microplus. The N- and P-domains of CRT were the most conserved, whereas the C-domain was high acid and more variable. CRT primary sequences were most conserved among mammals. Further investigations are warranted to determine whether immunization with Thai B. microplus CRT can affect tick performances and experimental pathogen transmission. PMID- 19120174 TI - Characterization of a Vitellogenin Gene Fragment in Boophilus microplus Ticks. AB - The objective of this study was to isolate, clone, and characterize a fragment of the vitellogenin (Vg) gene from a B. microplus tick strain from Mexico. Using cDNA and specific primers, an 1800-bp fragment was amplified, cloned, and transformed in into E. coli, and then sequenced. Comparative analysis with a previously reported sequence showed 99% identity at both the nucleotide and amino acid level. The predicted amino acid sequence of the Mexican Vg has 6 positive mutations. There is an insertion of an aspartic acid on position 26 and a deletion on position 552 with respect to the reported sequence. There were 11 predicted glycosylation sites conserved in both strains. It is concluded that there is a high sequence homology of Vg in both strains. PMID- 19120175 TI - Emergence and potential of high-throughput and integrative approaches in pathology. AB - In recent years a major revolution has occurred in the analysis and understanding of pathogenesis and host-pathogens/parasite interactions. This revolution has been achieved through the emergence of the high-throughput integrative approaches used in the "omics" fields-such as genomics, transcriptomics, proteomics, interactomics, and metabolomics. The novelty of these approaches has resulted from the development of high-throughput apparatus, assisted by the increasing power and software of computers that allow for high-speed, multifactorial simultaneous analysis of numerous samples. This level of integration allows for in-depth analysis of mechanisms, pace, and patterns of the evolution and adaptation of pathogens. This evolution from linear to multifactorial approaches has opened new ways of creating and characterizing new vaccines, diagnostic candidates, and drug targets. PMID- 19120176 TI - "Pathogeno-proteomics". AB - Many scientists working on pathogens (viruses, bacteria, fungi, parasites) are betting heavily on data generated by longitudinal genomic-transcriptomic proteomic studies to explain biochemical host-vector-pathogen interactions and thus to contribute to disease control. Availability of genome sequences of various organisms, from viruses to complex metazoans, led to the discovery of the functions of the genes themselves. The postgenomic era stimulated the development of proteomic and bioinformatics tools to identify the locations, functions, and interactions of the gene products in tissues and/or cells of living organisms. Because of the diversity of available methods and the level of integration they promote, proteomics tools are potentially able to resolve interesting issues specific not only to host-vector-pathogen interactions in cell immunobiology, but also to ecology and evolution, population biology, and adaptive processes. These new analytical tools, as all new tools, contain pitfalls directly related to experimental design, statistical treatment, and protein identification. Nevertheless, they offer the potency of building large protein-protein interaction networks for in silico analysis of novel biological entities named "interactomes," a way of modeling host-vector-pathogen interactions to define new interference strategies. PMID- 19120177 TI - Study of bovine trypanotolerance by whole transcriptome analysis. AB - African trypanosomiases are parasitic diseases transmitted by tse-tse flies, considered as the main sanitary obstacle to animal production development in sub Saharan Africa. However, if trypanosomiases have dramatic consequences on zebu (Bos indicus) populations, they have a weaker impact on the western African taurine (Bos taurus), which is known to be naturally tolerant to trypanosome infection. Mechanisms governing this trypanotolerant trait are still poorly understood, but today, recent postgenomic biotechnologies, such as the SAGE technique (serial analysis of gene expression) allow us to explore the full transcriptome. Twelve SAGE libraries were constructed from two trypanotolerant animals (N'Dama and Baoule) and one susceptible species of cattle (the Sudanese zebu) during an experimental Trypanosoma congolense infection; 43,458 different tags were obtained at several particular points during the infection (before infection, at the maximum of parasitemia, the maximum of anemia, and at the end of the experiment after value normalization). Bioinformatics analyses highlighted some interesting gene variations with respect to the trypanotolerance status of the animal. PMID- 19120178 TI - Mitochondrial DNA origin of indigenous malagasy chicken. AB - We report the mitochondrial DNA (mtDNA) characterization of 77 indigenous chickens (fighting and meat birds) from Madagascar, using DNA sequences of the first hypervariable segment of the D-loop. Comparison with reference samples from the African continent and Asia revealed two mtDNA haplogroups, suggesting a dual geographic and genetic origin for the indigenous Malagasy chickens. The most common haplogroup was present in 65 individuals of the two types; it is likely of Indonesian origin. The second haplogroup was observed in 12 fighting birds and meat chickens; it could be of African continental origin and/or the result of recent introgression with commercial lines. We further studied a G/A single nucleotide polymorphism at nucleotide position 1892 bp of the coding sequence of the Mx gene that is reported to be one of the candidate susceptible/resistant genes to viral infection in chicken. Our results indicate the "susceptible" allele G is the most common with frequencies of 65% and 70% in Malagasy fighting and meat chickens, respectively. However, the allelic frequency difference between the two types of chickens is not significant (P > 0.05). These results are discussed in light of our current linguistic and archaeological knowledge on the origin of indigenous Malagasy chickens. PMID- 19120179 TI - The effect of habitat fragmentation and species diversity loss on hantavirus prevalence in Panama. AB - Habitat fragmentation and diversity loss due to increased conversion of natural habitats to agricultural uses influence the distribution and abundance of wildlife species and thus may change the ecology of pathogen transmission. We used hantaviruses in Panama as a research model to determine whether anthropogenic environmental change is associated with changes in the dynamics of viral transmission. Specifically, we wanted to determine whether hantavirus infection was correlated with spatial attributes of the landscape at both large and small scales or whether these changes are mediated by changes in community composition. When analyzed at coarse spatial scales, hantavirus reservoirs were more commonly found in disturbed habitats and edge habitats than in forested areas. At local scales, reservoir species dominance was significantly correlated with the slope of the terrain. To evaluate the effect of small-mammal diversity loss on infection dynamics, we implemented an experiment with selective species removal at experimental sites. Seroprevalence of hantavirus was higher in the community of small mammals and increased through time in the experimental sites. The higher seroprevalence in experimental plots suggests that greater diversity likely reduces encounter rates between infected and susceptible hosts. Our studies suggest that habitat loss and fragmentation and species diversity loss are altering hantavirus infection dynamics in Panama. Our work represents a multidisciplinary approach toward disease research that includes biodiversity concerns such as environmental change and degradation, human settlement patterns, and the ecology of host and nonhost species, work that may be especially important in tropical countries. PMID- 19120180 TI - Recent discoveries of new hantaviruses widen their range and question their origins. AB - Hantaviruses belong to the Bunyaviridae family. While usually hosted by wild mammals, they are potentially pathogenic for humans, and several serologically distinct groups associated with different syndromes have been identified. Yet, investigations have mostly been conducted where human infections by hantaviruses constitute a real and well-identified public health problem, i.e., the holarctic and neotropical areas. Some hantaviruses have also been described from a Suncus murinus in India and a Bandicota indica in Thailand. In addition, recent investigations in Cambodia revealed new Hantavirus types. More recently, two new Hantavirus species were described: Sangassou from a Hylomyscus simus, and Tanganya from a Crocidura theresae, both from Africa (Guinea), thus strongly questioning the current views about geographic range, evolution, and epidemiology of hantaviruses. In such a framework, we have conducted a survey of Hantavirus diversity in Southeast Asia which allows us to isolate the Thailand virus and address questions about the taxonomy of their rodent hosts. Here we present a molecular analysis of representatives of all currently known Hantavirus species, thus allowing the comparison between the newly described ones with a large range sample of rodent hantaviruses. Our results clearly point to the presence of a particular lineage of hantaviruses in Southeast Asia. It also strongly suggests that new viruses, additional mammalian hosts and different related syndromes in humans are likely to be discovered in the near future, particularly in Southeast Asia and in Africa, where Muridae rodents are highly diversified. Furthermore, additional work is also urgently needed to investigate the hantaviruses associated with Crociduridae and Soricidae. PMID- 19120181 TI - Characterization of anaplasma infections in Sicily, Italy. AB - This study aimed to characterize infection with Anaplasma marginale, A. phagocytophilum, A. ovis, and A. platys in humans, animals, and ticks in Sicily, Italy, during 2003-2006. Serologic (competitive ELISA [cELISA]) and indirect immunofluorescence antibody [IFA]; N= 1990) and DNA (polymerase chain reaction [PCR]; N= 2788) tests were conducted on horse, donkey, cattle, sheep, goat, pig, dog, cat, roe deer, wild boar, human, and tick samples. The results reported herein suggested that in Sicily cattle are a major reservoir for A. marginale, dogs for A. platys, and sheep and goats for A. ovis. Domestic animals, such as cattle, horses, donkeys, sheep, dogs, and cats, may serve as reservoir for A. phagocytophilum, but different strains may infect ruminants and humans. All Anaplasma spp. characterized in Sicily had some distinctive genotypes for this region. Low genetic diversity was observed in A. ovis and A. platys, whereas A. marginale and A. phagocytophilum strains showed high genetic diversity. These results expanded our knowledge about the prevalence of Anaplasma spp. in Sicily and provided information to understand the epidemiology of these infections and implement measures to diagnose, treat, and control transmission to humans and animals in this region. PMID- 19120182 TI - Possible differential host tropism in Anaplasma phagocytophilum strains in the Western United States. AB - Despite organization into a single species, Anaplasma phagocytophilum, of bacteria from diverse hosts, there is evidence that different hosts vary in susceptibility to local strains of this bacterial pathogen. In particular, there is evidence that clinical hosts (humans, horses, and dogs) in the western part of the United States may be infected with strains of bacteria that differ from those in the reported wildlife reservoir, the dusky-footed woodrat. There is a lack of spatial overlap between clinical cases with woodrat exposure, genetic studies show highly distinctive strains in woodrats compared with those from clinical hosts, and experimental studies failed to transfer infection from woodrats into horses. These data suggest that multiple distinct A. phagocytophilum strains could circulate in western U.S. ecosystems. Host barriers to different bacterial strains would suggest the need for careful discrimination of A. phagocytophilum strains and further research to understand the host-pathogen interactions that result in differential outcomes in infection. PMID- 19120183 TI - Anaplasma marginale Yucatan (Mexico) Strain. Assessment of low virulence and potential use as a live vaccine. AB - Anaplasma marginale Yucatan strain was found to have low virulence in cattle. We studied the virulence of this isolate by experimental inoculation of 113 susceptible cattle at increasing doses, after which only one animal required treatment for clinical disease. Subsequently, 104 cattle received a live vaccine of this strain by inoculation, which induced immunoprotection after heterologous challenged exposure with a different A. marginale isolate. In this study 14% of the immunized cattle required treatment as compared with the control nonimmunized cattle, in which 56% required treatment. The A. marginale vaccine strains used for the immunization studies had MSP1a variable regions that were different from those used for the challenge exposure. PMID- 19120184 TI - Pathologic evidence of ehrlichiosis in calves inoculated with Ehrlichia chaffeensis. AB - An immunocompetent animal disease model based on infection with Ehrlichia chaffeensis would facilitate research toward understanding mechanisms responsible for the broad range of clinical signs associated with human monocytic ehrlichiosis (HME). The adaptability of this model for the experimental feeding of tick species and stages and for testing therapies comparable to those for human diseases are additional advantages of large animal models. Herein, we summarize pathology reports for calves that developed fatal disease after experimental inoculation with E. chaffeensis. Elevated liver enzyme levels and lung pathology among these calves corroborated earlier reports of severe HME. Thus, an experimental disease model based on infection of outbred immunocompetent hosts with E. chaffeensis could be within our grasp for the first time. PMID- 19120185 TI - First report of bovine anaplasmosis caused by Anaplasma centrale in Europe. AB - Anaplasma centrale (Rickettsiales: Anaplasmataceae) is used as a live vaccine for cattle against the pathogenic Anaplasma marginale in tropical and subtropical areas. Herein we report a clinical case of bovine anaplasmosis associated with A. centrale infection in Italy, together with the first molecular identification and phylogenetic analysis of this Anaplasma species or subspecies in Europe. PMID- 19120186 TI - Flow cytometry to evaluate Anaplasma marginale parasitemia using a fluorescent nucleic acid stain. AB - In this work we describe a flow cytometry-based method using SYTO16 (a DNA intercalating agent) to quantify Anaplasma marginale-infected erythrocytes in blood from bovine animals. The linearity and reproducibility of the results obtained with SYTO16 labeling followed by flow cytometry analysis make it a suitable approach for measurement of parasitemia in A. marginale infections. PMID- 19120187 TI - Immunophenotype of dogs with subclinical ehrlichiosis. AB - The purpose of this study was the characterization of the different subsets of lymphocyte cells in dogs with subclinical ehrlichiosis in order to contribute to the knowledge of the immune response developed in the course of this disease. Thirty-eight dogs with subclinical ehrlichiosis and 15 unaffected dogs from two shelters in the area of Valencia (eastern Spain) were included in the study. The study of lymphocyte populations was made by flow cytometry. Monoclonal antibodies against CD3, CD4, CD8, and CD21 were used. Based on our results, the most common findings of the subclinical phase of canine ehrlichiosis were lymphocytosis, relative neutropenia, and a decrease in the neutrophil/lymphocyte ratio. Lymphocytosis in these dogs was from an increase of T lymphocyte counts. Tc cell counts in dogs with subclinical ehrlichiosis were higher than in healthy dogs. This rise in the number of Tc lymphocytes resulted in a reduced percentage of Th lymphocytes and in a decrease in the CD4/CD8 ratio. PMID- 19120188 TI - Comparison between different polymerase chain reaction methods for the diagnosis of Ehrlichia canis infection. AB - The aim of this study was to compare different polymerase chain reaction (PCR) methods for the detection of Ehrlichia canis in blood samples and to relate these results to clinical findings and serology to E. canis using the indirect fluorescent antibody (IFA) test. Nine seropositive and nine seronegative dogs were included in this study. DNA was extracted once and used in one simple PCR and five nested PCR protocols previously described. In selected dogs (three seropositive and one seronegative) blood samples were aseptically collected in order to attempt the isolation of E. canis in the DH82 cell line. Results show that nested PCR protocols seem to be more sensitive than the simple PCR. Considering only nested PCR protocols, 33% of the IFA-positive samples were PCR positive using the five different protocols. The rest of the IFA-positive samples were PCR positive or negative depending on the protocol used. Clinical signs and laboratory findings compatible with canine monocytic ehrlichiosis (CME) were found in 67% of dogs positive by the IFA test. All samples positive by both techniques (IFA test and PCR) were from dogs suffering from clinical CME. IFA negative samples were PCR negative, except 22% that were PCR positive when using only one of the nested PCR protocols. Isolation of the agent was exclusively achieved in the only case in which the IFA test and all the PCR protocols were also positive. PMID- 19120189 TI - Phylogenetic analysis of Mexican Babesia bovis isolates using msa and ssrRNA gene sequences. AB - Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2), and msa-2b). Small subunit ribosomal (ssr)RNA gene is subject to evolutive pressure and has been used in phylogenetic studies. To determine the phylogenetic relationship among B. bovis Mexican isolates using different genetic markers, PCR amplicons, corresponding to msa-1, msa-2c, msa-2b, and ssrRNA genes, were cloned and plasmids carrying the corresponding inserts were sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 12 geographically different B. bovis isolates and a reference strain. Overall sequence identities of 47.7%, 72.3%, 87.7%, and 94% were determined for msa-1, msa-2b, msa-2c, and ssrRNA, respectively. A robust phylogenetic tree was obtained with msa-2b sequences. The phylogenetic analysis suggests that Mexican B. bovis isolates group in clades not concordant with the Mexican geography. However, the Mexican isolates group together in an American clade separated from the Australian clade. Sequence heterogeneity in msa-1, msa-2b, and msa-2c coding regions of Mexican B. bovis isolates present in different geographical regions can be a result of either differential evolutive pressure or cattle movement from commercial trade. PMID- 19120190 TI - Enhancement of the Mexican bovine babesiosis vaccine efficacy by using Lactobacillus casei. AB - To evaluate the effect of Lactobacillus casei on the effectiveness of the Mexican bovine babesiosis mixed vaccine, 20 bovines were randomly allocated into four groups of five animals (I, II, III, and IV). At day -2 animals in groups I and II were inoculated with saline solution by intramuscular route (i.m.) and animals in groups III and IV were inoculated with L. casei. At day 0 bovines in groups I and III were inoculated i.m. with bovine normal erythrocytes and animals of groups II and IV were inoculated with the babesiosis vaccine. Twenty-four days later each bovine was challenged with Babesia bovis- and B. bigemina-infected erythrocytes. The average rectal temperature in groups I and III was higher (P < 0.05) than that in the vaccinated groups after challenge. The average packed cell volume was lower (P < 0.01) in the control groups than in the vaccinated groups. At day 10 after challenge, the average anti-Babesia antibody level was higher in group IV than in group II. At day 7 after vaccination, the percentage of bovines positive to gamma interferon, as determined by real-time PCR, was 20, 0, 40, and 80 for groups I, II, III, and IV, respectively. All animals in control groups (I and III) were treated against babesiosis to avoid their death because they showed signs of babesiosis. The results indicate that L. casei, inoculated 2 days before the inoculation of the Mexican bivalent bovine babesiosis vaccine, improves the vaccine's efficiency. PMID- 19120191 TI - Infection and seroconversion of susceptible animals introduced into a babesiosis endemic area. AB - The study aimed to determine the incubation period of Babesia sp. infection in naive cattle and to monitor the serological response once exposed to natural Boophilus microplus (Rhipicephalus microplus)-infested paddocks. The study was carried out on a farm located in Veracruz, Mexico. Five groups of five steers were relocated every 3 months from a tick-free area to a tick-infested paddock. Animals were introduced in October, January, April, July, and October. Blood samples were taken daily until day 21 to determine packed cell volume (PCV), percentage of parasitized erythrocytes (PPE), and antibody titers to Babesia bigemina and B. bovis by the indirect fluorescent antibody procedure. Detection of Babesia in blood was also performed by species-specific PCR. The estimated incubation period was 6-14 days post introduction to paddocks (PIP), with fever (41 degrees C) for at least 3 days. PCV decreased by >25% and Babesia parasites were observed during the clinical phase of the disease. The highest individual PPEs (0.44% and 0.22% for B. bovis and B. bigemina, respectively) were observed from animals introduced in October. The four other groups showed a mean PPE ranging from 0.002-0.146% at day 14 PIP. All animals were detected as PCR positive between 8-14 days PIP. The highest antibody titers were 1:3328. The environmental conditions were favorable throughout the year for tick reproduction as the farm showed enzootic stability and hyperendemic conditions for bovine babesiosis. In this type of farm, strategic tick control could be accompanied by babesiosis vaccination, particularly for cattle relocated from tick-free areas. PMID- 19120192 TI - Expression analysis of heat shock protein 20 and rhoptry-associated protein 1a in sexual stages and kinetes of Babesia bigemina. AB - Heat shock protein 20 (HSP-20) and rhoptry-associated protein 1a (RAP-1a) are two proteins considered as candidates to be included in vaccines or diagnostics methods for the control of bovine babesiosis. It has been hypothesized that both genes have a basic function in the cellular physiology of erythrocyte-infecting stages; it is not known if they have a functional role in tick stages. The objective of this work was to analyze whether hsp-20 and rap-1a are expressed in sexual stages and kinetes of Babesia bigemina. Purified RNA from sexual stages and kinetes was analyzed by reverse transcriptase (RT)-PCR with specific primers for hsp-20 or rap-1a. Expression analysis was carried out using an indirect immunofluorescence test with specific antibodies against HSP-20 and RAP-1a. Results obtained by RT-PCR showed amplicons for hsp-20 and rap-1a in sexual stages and kinetes. Positive signals were also detected when sexual stages and kinetes were analyzed with specific antibodies for HSP-20 and RAP-1a. The results obtained here confirm the hypothesis that the genes hsp-20 and rap-1a from B. bigemina are expressed in sexual stages and kinetes and stress the importance of these proteins in the cellular physiology of tick stages. PMID- 19120193 TI - Molecular characterization of babesia bovis strains using PCR restriction fragment length polymorphism analysis of the msa2-a/b genes. AB - The merozoite surface antigen-2 (msa-2) family of Babesia bovis is a group of variable genes that share conserved 5' and 3' ends and encode for membrane anchored glycoproteins that have been postulated as vaccine candidates. In this work, we analyzed the sequences of three of these genes (msa-2a1, a2, and 2b) from two geographically distant strains and detected a certain degree of genotypic diversity that could be exploited to work out new molecular tools for the discrimination of B. bovis field samples. Here we describe a PCR restriction assay that was developed based on this observation and tested on several B. bovis strains and isolates. The results show a strain-specific band pattern in geographically distant isolates, indicating the presence of differentially located BspMI restriction sites. This approach provides a simple method for the differentiation of American B. bovis strains. PMID- 19120194 TI - msa-1 and msa-2c gene analysis and common epitopes assessment in Mexican Babesia bovis isolates. AB - Babesia bovis msa-1 and msa-2c genes belong to the variable merozoite surface antigen gene family. These genes code for antigenic proteins present on the merozoite surface (MSA) and are involved in the parasite invasion to the bovine erythrocyte. Previous studies carried out on MSA-1 have evidenced antigen allelic variation in B. bovis isolates from similar endemic regions, as well as in isolates from different geographic regions of the world (Argentina, Australia, Israel). Studies conducted on MSA-2c, however, have shown that this antigen is widely conserved on isolates from distinct geographic regions. In this study, it was hypothesized that MSA-1 and MSA-2c antigens would contain common epitopes despite the presence of nucleotide sequence differences found in 13 B. bovis isolates and strains collected in geographically distant regions of Mexico. Bioinformatics analysis of the primary structure from DNA fragments derived from PCR amplification, cloning, and sequencing of msa-1 and msa-2c genes from the 13 B. bovis populations revealed that the msa-1 gene product present in the various isolates tested is less conserved among isolates obtained within a similar geographic region in Mexico (51-99.7% sequence identity). Results obtained by immunoblot analysis of B. bovis protein extracts reacted with a monoclonal antibody to MSA-1 42-kDa antigen, conclusively showed cross-reactive common epitopes only in Mexican isolates having high sequence identity (>/=99%, eight isolates). Sequence analysis and multiple alignment of deduced MSA-2c demonstrated a high degree of sequence identity (90-100%) among the Mexican B. bovis isolates and strains. Immunoblot results using a polyclonal antibody to MSA 2c reacted against the protein extracts recognized conserved epitopes in at least nine of the B. bovis isolates. The results obtained in this study agree with those previously reported by other researchers and confirm that, based in sequence identity conservation in Mexican B. bovis isolates and strains so far collected and analyzed, MSA-2c represents an ideal antigen worth evaluating as a vaccine candidate. PMID- 19120195 TI - Water Buffalos as carriers of Babesia bovis in Argentina. AB - The tick-transmitted hemoprotozoan Babesia bovis is a major causative agent of bovine babesiosis, an often fatal disease of cattle. The disease is widespread in the northeastern region of Argentina, where an increasing part of the livestock is composed of water buffalos. Although clinical cases of buffalo babesiosis have not been reported so far, the pathogen-transmitting tick vector has been occasionally observed by us to be feeding on water buffalos. We therefore set out to examine whether buffalos may constitute a reservoir of the parasite. Competitive enzyme-linked immunosorbent assay (cELISA) detected B. bovis-specific antibodies in 20% of investigated buffalos (21/103), while direct detection of the pathogen by nested PCR was demonstrated in 34% of the animals (35/103). In one field, more than 60% of investigated animals (22/36) tested positive by nested PCR. These results are discussed in the context of buffalo babesiosis reported in other countries and in view of the currently effected control measures against bovine babesiosis in the region. PMID- 19120196 TI - Effect of high- and low-molecular-weight components of Lactobacillus casei on resistance against Babesia microti in NIH mice. AB - Three assays were carried out in order to assess the effect of low- and high molecular-weight components (LMWc and HMWc, respectively) of Lactobacillus casei on the resistance of mice to Babesia microti infection. In the first study, 23 components were identified in a total extract of L. casei by SDS-PAGE. In the second experiment, 15 components (ranging from 19 to 148 kDa) from L. casei (viable) and 12 other components (from 21 to 148 kDa) from L. casei (dead) were recognized by the sera of immunized mice in the extracts of sonicated L. casei. On the basis of these results, samples of sonicated L. casei were separated by preparative gel electrophoresis, and then those gel fragments containing the HMWc (range 63 to 111 kDa) and LMWc (range 19 to 59 kDa) were cut and eluted to constitute the LMWc and HMWc of L. casei. In the third study, five groups of 6 mice each received the following treatments: PBS in the control (C) group, L. casei LMWc in the LcL group, L. casei HMWc in the LcH group, total sonicated L. casei (LMWc + HMWc) in the LcT group, and viable L. casei in the LcV group. At day 0 each mouse in all groups was challenged with 2 x 10(4)B. microti parasitized erythrocytes. At day 8 after challenge, the average percentage of parasitized erythrocytes +/- SEM was significantly higher (P < 0.05) in the C group (11.8 +/- 0.9) as compared with LcL (7.7 +/- 1.5), LcH (10.3 +/- 6.9), LcT (3.6 +/- 0.7), and LcV (6.9 +/- 1.9) groups. The results suggest that L. casei LMWc can induce an early protective immune response, similar to that generated by LcV in mice, against B. microti infection. PMID- 19120197 TI - Improved molecular tools for detection of Babesia bigemina. AB - Molecular detection of Babesia bigemina involves a nested PCR protocol and reverse line blot hybridization (RLBH) assay based on the 18S gene. In this study, we report the development of molecular tools for improving B. bigemina detection in bovine blood-a one-step PCR assay based on the amplification of rap 1a paralogous and a new RLBH Babesia spp. 18S probe. The one-step PCR assay is highly specific, with an estimated analytical sensitivity corresponding to 0.00002% parasitemia. The RLBH assay, with a new B. bigemina probe, allows the detection of all tested B. bigemina isolates showing no cross-hybridization with B. bovis 18S gene. By developing this highly specific and sensitive one-step PCR and upgrading the RLBH assay for B. bigemina, we have improved molecular assays which, together with serologic methods, provide valuable tools for epidemiologic studies of bovine babesiosis. PMID- 19120198 TI - In vitro nematocidal activity of plant extracts of Mexican flora against Haemonchus contortus fourth larval stage. AB - Fourteen plant extracts were evaluated in vitro against the fourth larval stage of Haemonchus contortus. The plants species used were Tagetes erecta, Argemone mexicana, and Castela tortuosa. The assays were run in 24-well cell culture plates at room temperature with three replicates. After exposure, aliquots were taken from the corresponding wells and transferred to a microscope for observation. Evaluation criteria were based on the average of live and/or dead larvae. ANOVA test and Tukey test were used to determine significant differences among the treatments. After 96 h, the T. erecta acetonic extract produced 99.7% lethal activity, followed by C. tortuosa hexanic extract (85.8%) and T. erecta methanolic extract (58.3%) (P < 0.0001). PMID- 19120199 TI - Evaluation of multinutritional pellets containing Duddingtonia flagrans chlamydospore for the control of ovine haemonchosis. AB - This research evaluated the use of multinutritional pellets (MP) containing Duddingtonia flagrans chlamydospores as vehicles for oral administration in sheep against Haemonchus contortus. Pellets containing urea, wheat coat, soy meal, molasses, and D. flagrans chlamydospores were offered for 10 days to six nematode free sheep (group 1, treated). Group 2 (control), also six nematode-free sheep, was supplied for the same period with nonfungi pellets. Feces were collected daily from each animal and deposited on water agar plates containing 500 H. contortus infective larvae. Three plates were used as three replicates per treatment, and all plates were incubated for 7 days. Microscopic observations revealed trap formation and the captured larvae. Washes from the plate content were passed through a Baermann funnel to collect noncaptured larvae and to quantify nematode reduction by fungal action. A larval reduction of 82% was found in fungal culture plates. The use of MP containing D. flagrans chlamydospores could be considered as a viable alternative control tool against sheep haemonchosis. PMID- 19120200 TI - Lethal activity of two Bacillus thuringiensis strains against Haemonchus contortus histotropic larvae. AB - The in vitro lethal activity of two Bacillus thuringiensis strains (IB-16 and IB 61) against Haemonchus contortus histotropic fourth stage larvae (L(4)) was evaluated. Each B. thuringiensis strain was used as soluble proteins at different concentrations. In addition, the in vitro lethal dose (LD(50)) of each strain was determined against H. contortus L(4) at different times. Data were analyzed by ANOVA and Tukey tests. Less than 50% mortality was observed at 50 and 100 microg/mL during all bioassays. More than 50% mortality was observed at 200 microg/mL at 3 and 5 days after interaction with IB-16 and IB-61, respectively. A 100% mortality was observed with IB-16 on day 5. The LD(50) with IB-16 was 248 microg/mL on day 3, and IB-61 showed an LD(50) at 227 microg/mL on the day 5. Both toxins may have potential as biocontrol agents. PMID- 19120201 TI - The little-known scenario of anthelmintic resistance in equine cyathostomes in Italy. AB - Cyathostomes (Nematoda, Cyathostominae) cause colic, decreased performance and growth, peripheral edema, and dysorexia in equids. Also, the synchronous emergence of cyathostome larvae encysted in the colon and cecal wall causes larval cyathostominosis, which is characterized by protein-depriving enteropathy, chronic diarrhea, edema, weight loss, colitis, and may be fatal. In the past few decades, drug resistance in cyathostomes has become widespread, especially for benzimidazoles. Resistance to tetrahydropyrimidines is still confined to a few areas, and the recent reports of reduced efficacy in macrocyclic lactones are of great concern. Although cyathostomes are widely distributed in Italy, with a prevalence up to 100%, information on the presence of drug-resistant cyathostomes is scarce. The first study investigating drug resistance was conducted on a few farms in 1992 and showed the presence of cyathostome populations resistant to the (pro)benzimidazoles in northern Italy. A second survey was carried out 14 years later, with the focus on farms in central and southern regions. Cyathostomes resistant to fenbendazole and pyrantel pamoate were found, while macrocyclic lactones remained 98-100% effective. Recently, the efficacy of oxibendazole (97%) and moxidectin (100%) in removing fenbendazole-resistant cyathostomes in Italy has also been demonstrated. These few studies indicate that drug-resistant cyathostomes are present in multiple regions of Italy, but geographically and numerically broader investigations of farms are necessary. Finally, there is the undoubted need to use the remaining fully effective drugs (such as moxidectin) in a proper way to preserve their efficacy as much as possible. PMID- 19120202 TI - Cytokine response in the intestinal mucosa of hamsters infected with Taenia solium. AB - Taenia solium grows in experimentally infected hamsters. An inflammatory reaction in the intestinal mucosa surrounding the scolex of the worms is produced. We searched for mRNA of Th1 and Th2 cytokines by in situ hybridization in intestinal biopsies. Hamsters were infected with T. solium cysticerci and necropsied on different days post infection (d.p.i.). Tissue from the small intestine was taken from the area surrounding the tapeworm scolex, fixed, and processed for histology. Antisense probes for the detection of interferon (IFN)-gamma, interleukin (IL)-4, IL-5, and IL-13 were used. Kinetics of each cytokine was defined through detection on specific mRNA by counting the number of positive infected hamsters and of positive cells per 100 enterocytes on different d.p.i. IFN-gamma was detected as of d.p.i. 2; all animals were positive on d.p.i. 4 and 8; and on d.p.i. 16, only 20% were still positive. IL-13 had a pattern similar to IFN-gamma, but all hamsters remained positive until d.p.i. 16 when the experiment was terminated. IL-4 was positive in 40% of infected hamsters on d.p.i. 6. On d.p.i. 8, IL-5 was only detected in 20% but increased to 100% by d.p.i. 16. These data suggest that tapeworms induce a mixed Th1/Th2 response with a polarization toward Th2 at 2 weeks post infection, which may influence the expulsion of worms. PMID- 19120203 TI - New epidemiological and molecular insights into feline lungworm infection. AB - Aelurostrongylus abstrusus (Nematoda, Strongylida) lives in the terminal respiratory bronchioles and alveolar ducts of cats. The infection may present with respiratory signs of varying severity and can be life threatening. Despite the importance of A. abstrusus in feline medicine and recent reports of clinical cases of aelurostrongylosis in Italy, epidemiological data are sparse and molecular information available on the parasite are lacking. The present work reports recent insights into the epidemiology of feline lungworm infection in Italy and contributes to the molecular knowledge of A. abstrusus. Individual stool samples were collected from 100 animals and examined with flotation and Baermann methods. The risk factors were analyzed by binary logistic multiple regression models. All positive fecal samples (n= 16) were subjected to the PCR coupled sequencing of the rDNA internal trancribed spacer 2 (ITS2). The detected infection rate differs significantly from data reported in the literature, and stray and free-ranging cats and the presence of respiratory symptoms were risk factors for the infection. The molecular characterization showed that the ITS2 sequence is a promising genetic target for a holistic approach to the study of cat lungworm infection. Epidemiological and molecular insights and avenues are discussed. PMID- 19120204 TI - Phenotype characterization of Pelibuey native lambs resistant to Haemonchus contortus. AB - Specific phenotypic traits of the blood-feeder nematode Haemonchus contortus were characterized on Pelibuey native, cross-breed sheep in a tropical region of Mexico with background of anthelmintic problems. Data were collected on 4-month old lambs (n= 63) and analyzed weekly in three stages: stage 1 (S1, not infected (0-3 weeks)); S2, first infection with 350 H. contortus infective larvae (L(3)) per kg of body weight by oral route (3-10 weeks); and S3, second infection with 175 H. contortus L(3) per kg of body weight by oral route (10-18 weeks). Once infection took place, sheep grazed in a free-nematode experimental paddock, and hemoncosis-related traits, such as the number of eggs per gram of feces (epg), packed cell volume (%pvc), and eosinophils (%eo), were measured each week. Results were analyzed on specific traits and stages using SAS statistics. Statistical differences of the main phenotypic traits (P < 0.05) were observed throughout S2 on Pelibuey high (hr) and low (lr) responders, respectively: epg mean, 475 +/- 340.5 and 1439 +/- 243.6; %pvc mean, 31 +/- 0.3 and 27 +/- 0.2; and %eo mean, 2.5 +/- 0.3 and 1.99 +/- 0.2. In addition, lr and hr sheep grazed for 3 months on an area infected with Haemonchus, Cooperia, and Strongyloides, and the hr sheep showed a low number of epg after being exposed to multiple infections, so that no further treatment was required. These results showed that the selection of animals resistant to nematodes can be an alternative method of control against anthelmintic resistance problems in order to improve animal health and avoid a negative environmental impact. PMID- 19120205 TI - In vitro fasciolicide activity of some plant extracts against newly excysted flukes. AB - The aim of the present study was to evaluate, under in vitro conditions, the activity of 20 plant extracts against newly excysted Fasciola hepatica metacercariae. The plants were selected on the basis of anthelmintic activity, traditional use, availability, solubility, and permanence in the liver. For the in vitro evaluation, 24-well Nunc-immunoplates containing RPMI-1640 culture medium bovine sera plus antibiotics were used. Extract concentrations of 500 and 50 mg/mL were placed in triplicate. Approximately 10 newly excysted flukes were placed in each well, and the plates were incubated in a CO(2) incubator at 5% atmosphere for 3 days. In each plate four control wells containing nontreated flukes were included. All procedures were undertaken under an air-flow cabinet. Efficacy was measured on days 1 and 3 by considering the motility of the live or dead trematodes evaluated with the aid of an inverted microscope. Results indicated that, from all extracts evaluated, Tilia mexicana showed an average efficacy for concentrations of 500 and 50 mg/mL, respectively, of 100 and 100%; Eriobotrya japonica, 100 and 95%; Coffea arabica, 100 and 100%; Crataegus mexicana, 95 and 90%; and Ternstroemia pringlei 100 and 100%. From this preliminary evaluation, we conclude that further and more detailed evaluations at lower concentrations should be undertaken to understand the real efficacy of these plant extracts. PMID- 19120206 TI - In silico fasciolicide activity of three experimental compounds in sheep. AB - The aim of the present study was to evaluate the fasciolicide activity of three experimental drugs, selected by an in silico system called TOMOCOMD-CARDD, in sheep. Drugs were identified by the computer system, and, after statistical selection, 24 Pelibuey sheep were infected on days 0 and 30, each with 200 metacercariae of Fasciola hepatica. When the infection reached 8 and 4 weeks of age, respectively, four groups of six animals each were formed. Group 1 received thiacetazone 150 mg/animal/p.o. Group 2 was treated with 3,5,5, trimethyloxazolidine 2,4-dione at 450 mg/animal/p.o. G3 received guanabenz acetate at a dose of 1.5 mg/animal/p.o. G4 served as an untreated control. Monitoring of the animals was followed by individual coprological examinations and slaughter of the animals 15 days after treatment to collect and count flukes from the liver. Efficacy was measured as the reduction in the percentage of flukes of treated animals relative to untreated controls. Results indicated an efficacy of 80.0, 43.8, and 100% for 8-week-old flukes and 62.1, 57.9, and 100% for 4-week-old flukes in the three experimental groups, respectively. Even though guanabenz acetate showed a high efficacy, it was highly toxic since two animals died approximately 24 h after being treated. We conclude that further investigations should be conducted to perform computer-aided prediction of drugs aimed to detect fasciolicide activity. PMID- 19120207 TI - Immune response of Theileria sp.-infected sheep to recombinant Theileria proteins. AB - Sheep and goats in northwest China suffer from theileriosis from infection with Theileria sp. (China), resulting in large economic losses. To investigate the immune response to infection with Theileria sp. (China), parameters of cellular and humoral immunity of experimentally infected sheep against two recombinantly expressed Theileria proteins were investigated. The in vitro proliferative response of blood mononuclear cells to a recombinant T. annulata membrane protein and a recombinant Theileria sp. (China) homologue to T. annulata surface protein, both putative membrane proteins, was significantly elevated and significant amounts of specific immunoglobulins were produced against both. PMID- 19120208 TI - Amblyomma variegatum ticks and heartwater on three Caribbean Islands. AB - Amblyomma variegatum tick infestation, tick infection by Ehrlichia ruminantium (ER), and ER genetic diversity were studied in the Caribbean Islands of Guadeloupe, Marie-Galante, and Antigua between 2003 and 2005. Nested PCR for pCS20 was used to detect ER, while ER strains were characterized by sequencing or by restriction fragment length polymorphism (RFLP) profiles of map-1 PCR products. In 2003 in Guadeloupe, the prevalence of tick-infested herds was 35.6%. In Marie-Galante 79.1% of herds in 2003 and 73.8% in 2005 were infested, while only an average of 2.2% of the herds were infected in Antigua between this same period. In Marie-Galante, 19.1% of ticks were ER positive, and ER-infected ticks were found in 33.3% of the herds. In Antigua only 5.8% of the ticks were ER positive. High ER tick infection rate combined with a very high level of tick infestation highlight the risk of heartwater in Marie-Galante and Guadeloupe more than in Antigua. The three islands still represent a reservoir for tick and heartwater in the Caribbean. Nine different African and Caribbean map-1 ER genotypes were identified. This diversity was observed even in restricted areas, and identical map-1 genotypes were observed on all three islands. This high genetic diversity of ER strains suggests that there was a simultaneous introduction of several strains from African countries into the Caribbean region. PMID- 19120209 TI - The application of PCR and reverse line blot hybridization to detect arthropod borne hemopathogens of dogs and cats in Trinidad. AB - Arthropod-borne diseases are important causes of morbidity and mortality of companion animals in Trinidad. As clinical signs are vague, more sensitive methods to diagnose these diseases based on the polymerase chain reaction (PCR) followed by reverse line blot hybridization (RLB) of amplified products are being developed. An RLB of 14 oligonucleotide probes coupled with polymerase chain amplified regions of 16S rRNA or 18S rRNA genes of hemoparasites from cats and dogs detected Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, feline mycoplasmas ("Candidatus Mycoplasma haemominutum,"Mycoplasma haemofelis), and some unknown species within the Babesia/Theileria group and the Anaplasma/Ehrlichia tribe. Amplified products were obtained from blood samples collected from 348 dogs and 15 cats. Overall, hemopathogen DNA was detected in 92 (26.4%) dogs and six (40.0%) cats. E. canis (49, 14.1%) and feline mycoplasma (5, 33.3%) DNA were most frequently identified in dogs and cats, respectively. B. canis vogeli (1, 6.7%) and E. canis (1, 6.7%) were also detected in cats. Mixed infections of Anaplasma/Ehrlichia sp. and Babesia sp. were present in five (1.4%) dogs, while mixed infections of the feline mycoplasmas were present in two (13.3%) cats, one of which was also positive for E. canis. Pyrexia was significantly associated with a positive RLB result in dogs (P= 0.00, chi(2), 1 df). This is the first reported application of macro-arraying techniques to detect arthropod-borne hemopathogens of companion animals in the Americas and the first detection of DNA of B. canis vogeli and E. canis in cats in Trinidad. PMID- 19120210 TI - Immunization with tick salivary gland extracts. AB - Tick burdens cause direct damage to hosts and transmit several disease agents, the majority of which are secreted into feeding lesions through tick salivary glands. Reduced incidence of naturally transmitted tick-borne diseases was recently observed among cattle immunized with tick salivary gland extracts (TSGE). The aim of this work was to compare the ultrastructure of salivary glands from Rhipicephalus (Boophilus) microplus removed from cattle that were immunized with TSGE to those from control cattle injected with saline. Partially fed female ticks that infested these cattle were collected, and their salivary glands were removed and compared with light microscopy. More extensive salivary gland damage was observed in ticks collected from cattle immunized with TSGE than those from saline controls. Salivary glands from immunized cattle had extensive necrosis in agranular c and f cells; partial necrosis in granular b, c, and e cells; and slight necrosis in a and d cells. These results indicated that host immunization with TSGE causes detrimental effects to female tick salivary glands and correlated with the reduced incidence of naturally transmitted tick-borne diseases observed among these cattle. PMID- 19120211 TI - Study on Theileria lestoquardi antigens as potential vaccine candidates. AB - Theileria lestoquardi is the causative agent of malignant theileriosis of sheep and goats, causing morbidity and mortality in these animals worldwide. Western blot analysis based on T. lestoquardi schizont antigens was carried out using sera collected from Iranian sheep, which had been immunized with T. lestoquardi schizont-infected cells. The results of Western blot analysis demonstrated that schizont-immunized animals produced antibodies reacting with protein bands at 73, 42, 20, 14, and 12 kDa. Comparison of the results of the current Western blotting test with earlier studies of Theileria spp. revealed two immunogenic schizont proteins with molecular weights of 73 and 42 kDa shared between T. annulata and T. lestoquardi. Two other proteins with molecular weights of 14 and 12 kDa have not been previously found in other Theileria species. Our results suggest that the 73-kDa protein could be a potential vaccine candidate and that the 14- and 12 kDa proteins could be considered as diagnostic antigens. PMID- 19120212 TI - Field collection and genetic classification of tick-borne Rickettsiae and Rickettsiae-like pathogens from South Texas: Coxiella burnetii isolated from field-collected Amblyomma cajennense. AB - We are reporting the first known isolation of the Q-fever agent Coxiella burnetii from field-collected cayenne ticks Amblyomma cajennense in North America. Q-fever affects a number of domestic ungulates where it can lead to abortion in sheep and goats. There is far less known about the disease's effects on wild species, primarily because of the tendency of the disease to self resolve and to provide long-term immunity to subsequent infections. The first recovery of C. burnetii in North America was from the tick species Dermacentor andersoni. Since the original isolation C. burnetii has been recovered from five other North American tick species. The currently accepted mode for the majority of human infections is inhalation. The Centers for Disease Control and Prevention, Division of Viral and Rickettsial Diseases, Rickettsial Zoonoses Branch asserts the Q-fever agent as requiring as few as one organism to cause disease via inhalation in susceptible humans. However, with more and more isolations from ticks, evidence linking C. burnetii and ticks is mounting. The true role of tick species as competent vectors is still unconfirmed. Preemptive field collections of possible vector arthropods, hosts, and reservoirs can provide invaluable baseline environmental data that will prove supportive in follow-up studies and abatement efforts. PMID- 19120213 TI - Existence of splicing variants in homologues of Theileria lestoquardi clone-5 gene's transcripts in Theileria annulata and Theileria parva. AB - Clone 5 has been described as an immunogenic protein and was used to establish an ELISA for malignant theileriosis. Molecular characterization of the gene product revealed alternative splicing at the single intron resulting in two mRNA transcripts, translating into a long and a short protein form. Homologues of clone 5 exist in Theileria annulata and T. parva according to the available annotated GenBank sequences, showing however only the long protein forms in these parasites (GenBank accession numbers CAI73679, EAN33624). The present study aimed to determine whether two splice variants of homologues of clone 5 occur in T. annulata and T. parva. PMID- 19120214 TI - Small-scale expressed sequence tag analysis of Theileria uilenbergi: identification of a gene family encoding potential antigenic proteins. AB - Recently, Theileria sp. (China) has been designated as T. luwenshuni[formerly Theileria sp. (China 1)] and T. uilenbergi[formerly Theileria sp. (China 2)]. A cDNA library of T. uilenbergi merozoites was constructed and subjected to random sequencing. Among the obtained sequences were three highly identical cDNA clones, indicating a gene family. Bioinformatic analyses indicated these genes contain signal peptides and encode potential immunogenic proteins. The presence of tandemly arranged and additional variants of these genes was shown. Analysis of one recombinantly expressed clone revealed immunoreactivity for serum from Theileria-infected animals. No cross-reaction with serum of T. lestoquardi-, Babesia motasi-, or Anaplasma ovis-infected animals was observed, indicating a potential antigen for development of serological diagnostic tools. PMID- 19120215 TI - Comparison of the molecular structure of the TaSP gene of Theileria annulata from Sudanese isolates. AB - The polymorphic region of the Theileria annulata surface protein (TaSP) was cloned and sequenced from different isolates of cattle and cell lines from different areas of Sudan. Amino acid sequence alignment revealed a high diversity showing amino acid and length polymorphism, both within and between parasite isolates. The generation of TaSP diversity may allow the evasion of host immunity by the parasite since TaSP is a highly antigenic parasite protein. PMID- 19120216 TI - Cutaneous basophilia in the resistance of goats to Amblyomma cajennense nymphs after repeated infestations. AB - The acquisition of resistance in goats against Amblyomma cajennense after repeated infestations and the role of inflammatory cells in this mechanism were investigated. Ten naive goats aged 6 months were distributed into two groups: test (n= 5), infested thrice at 30-day intervals; and control (n= 5), infested once. Nymphs (n= 115 per animal per infestation) were released inside alimentary chambers glued to the animal's dorsum and the following biological parameters were evaluated: yielding rate, nymphal engorgement weight, engorgement period, ecdise rate, and ecdise period. Skin fragments of tick bite sites were collected at 24, 48, 72, and 120 h post attachment for histopathology and inflammatory cells counts. The engorgement weight decreased significantly (P < 0.05) from the first infestation onward; nymphs weighed 41.7% and 37.1% less after the second and third infestations, respectively, as compared to those collected after the first infestation (12.55 mg +/- 3.21). Furthermore, the ecdise period increased significantly (P < 0.05) in the third infestation (18 days +/- 2.83) in comparison with the first infestation (15 days +/- 0.82) and the ecdise rate was significantly lower after the second infestation (71.91%+/- 17.38) in comparison with the first infestation (96.89%+/- 3.38). There were no significant differences with regard to both the engorgement period and yielding rate. A cutaneous basophilia was evidenced between 48 and 72 h (P < 0.05) after both the second and third infestations. We conclude that goats develop resistance against nymphs of A. cajennense and that basophils may play an important role in such a mechanism. PMID- 19120217 TI - Guinea pigs develop cutaneous basophilia after repeated infestations by nymphs of the tick Amblyomma triste. AB - This study investigated the development of resistance in guinea pigs to nymphs of Amblyomma triste ticks after repeated infestations. Guinea pigs were infested thrice, at 30-day intervals, with 30 nymphs of A. triste per animal per infestation. Acquisition of resistance was evaluated by determining: nymph yielding rate, engorgement period, and weight. Skin biopsies of tick bite sites were collected at 24, 48, and 96 h after tick attachment for inflammatory cell counts. Engorged nymphs weighed 5.53 mg +/- 1.9 in re-infested hosts (56.6% less than in primary infestation) and took 6.9 days +/- 2.16 to feed in the third infestation (14.5% more than in the first infestation). Guinea pigs yielded 78%+/ 7.2 of nymphs in the re-infestation (11.6% less than in the primary infestation). In addition, a marked increase in basophil influx was observed from the first infestation onward; 69% of infiltrated cells were basophils in re infested hosts 24 h after tick attachment. No basophils were seen at this time in primary infested animals. That number increased to 84.7% 48 h post attachment in re-infested hosts (73.2% more than in the primary infested ones) and decreased markedly 96 h post attachment in both groups (2.6% and 1% of basophils in the third and first infestation, respectively). We conclude that guinea pigs acquire resistance against nymphs of A. triste ticks after repeated infestations and that a cutaneous basophilia must play a crucial role in such a mechanism. PMID- 19120218 TI - Effect of Amblyomma cajennense ticks on the immune response of BALB/c mice and horses. AB - This work evaluated the effect of the Amblyomma cajennense tick on the immune response of BALB/c mice and on horse lymph node cell proliferation. We observed that mice do not develop resistance to nymphs of this tick species and that lymphocyte proliferation of this host is inhibited by tick saliva, nymphal extract, or infestations. Horse lymph node cell proliferation is inhibited by tick saliva as well. Mice lymphocytes under the effect of tick saliva, nymphal extract, or infestations display a predominantly Th-2 cytokine production pattern. Observed results partially explain this tick's disease vectoring capacity and broad host range. PMID- 19120219 TI - Development of an indirect ELISA for the diagnosis of equine piroplasmosis. AB - An indirect ELISA (iELISA) for the detection of specific anti-Theileria equi antibodies in horse serum was developed. Its performance showed good concordance (K= 0.79) when compared with a competitive ELISA recommended by the World Organisation for Animal Health. Horse serum samples from two provinces located in the north and east of Argentina (Formosa and Entre Rios, respectively) were analyzed by this iELISA. A high percentage of positive horses were found in Formosa, consistent with the climatic conditions of the region that are apt for the development of tick vectors. Surprisingly, seropositive animals were also detected, although in a lower proportion, in Entre Rios, which has a temperate weather and is presumably tick free. Breeding of thoroughbred racing horses is an important economic asset of Argentina. Since equine piroplasmosis is a constraint for horse export, the epidemiological situation in different regions of the country needs to be assessed for the implementation of control measures. The iELISA developed in this work provides a convenient tool for this type of study. PMID- 19120220 TI - Fungal formulations to control Rhipicephalus sanguineus engorged females. AB - Entomopathogenic fungi are promising alternatives to chemical acaricides in controlling tick populations; however, biological formulations are required to improve conidial performance of these pathogens in the field. This study evaluated the effect of in vitro treatment of Rhipicephalus sanguineus with conidial formulations of Beauveria bassiana (Bb) or Metarhizium anisopliae (Ma). Conidia were formulated (at 10(8) conidia mL(-1)) in (1) emulsifiable concentrate (EC), (2) polymerized cellulose gel (PCG), or both (3) emulsifiable concentrate and polymerized cellulose gel combined (EC+PCG). Engorged females were weighed and exposed to one of six treatment groups: EC+Bb, EC+Ma, PCG+Bb, PCG+Ma, EC+PCG+Bb, EC+PCG+Ma; or to control groups: C1 (untreated) or (C2) treated with Tween 80 aqueous solution, EC or PCG. After exposure, ticks were placed in petri plates and kept at 27 +/- 1 degrees C and 80% relative humidity (RH). Ticks treated with formulated conidia of entomopathogenic fungi showed significantly higher control percentage in comparison to that of control groups. The formulation of B. bassiana with EC and PCG combined (EC+PCG+Bb) caused the highest control percentage (86.79%) of R. sanguineus. This study concludes that entomopathogenic fungus conidial formulations are important to ensure effective tick control. PMID- 19120221 TI - Performance of Metarhizium anisopliae and Its combination with deltamethrin against a pyrethroid-resistant strain of Boophilus microplus in a stall test. AB - The use of the entomopathogenic fungus, Metarhizium anisopliae, to control ticks is a promising alternative to chemical acaricides, but both chemical and biological products could also work together in an integrated management. The current study evaluated the performance of M. anisopliae and its combination with deltamethrin to control a Boophilus microplus strain resistant to pyrethroid. Calves were held in individual pens and artificially infested with B. microplus. They were treated with an aqueous conidial suspension of M. anisopliae, a deltamethrin solution, or both fungus and deltamethrin combined. After treatment, engorged females that naturally dropped off from calves were counted daily and evaluated in the laboratory. The mean mortality rate was 32.57% in the groups treated with M. anisopliae, 38.58% in those groups treated with deltamethrin, and 30.92% in those treated with fungus and chemical combined. Additionally, ticks that dropped off from calves after 2 days had a lower nutrient index (NI) and egg production index (EPI) than those that dropped off at a later day after treatment. This indicates that M. anisopliae may reduce B. microplus population only for a few days after treatment when a simple aqueous conidial suspension was used. PMID- 19120222 TI - In vitro efficacy of amitraz and cypermethrin on Boophilus microplus from dairy farms in Lara State, Venezuela. AB - In Venezuela, the acaricides, amitraz and cypermethrin, are widely used to control tick infestations in cattle infested with Boophilus microplus. This work aims to determine the in vitro efficacy of amitraz and cypermethrin through the adult immersion test. A sample of 480 engorged females was collected from four dairy farms (180 ticks/farm) and divided in groups of 10 ticks averaging 220 mg/tick. Commercial preparation of amitraz and cypermethrin were used to prepare the working concentration of amitraz (52, 104, 208, 416 and 832 ppm), and cypermethrin (37.5, 75, 150, 300, 600 ppm). The control groups were submerged only in water. Ticks were placed in a glass lamina at 27 degrees C and 85% relative humidity (RH) and incubated for 18 days. The eggs masses were collected, weighed, separated in vials, and allowed to hatch at 85% RH and 27 degrees C for 21 days. Both the emerged larvae and eggs were quantified in each tube in order to determine the efficacy percentage and the resistance factor. The efficacy average to amitraz was 30.89%, 50.75%, 50.62%, 59.72%, and 60.53% at concentrations of 52, 104, 208, 416, and 832 ppm, respectively. The efficacy of cypermethrin was 14.3%, 11.82%, 17.25%, 20.23%, and 43.23% at concentrations of 37.5, 75, 150, 300, and 600 ppm, respectively. The amitraz resistance factor to each farm was 3.14, 1.19, 1.88, and 1.89, whereas with cypermethrin it was 8.89, 6.03, 7.12, and 18.45. Statistically significant differences between the effectiveness of different concentrations of amitraz or cypermethrin were not observed. Higher concentrations do not increase the efficacy of the products. PMID- 19120223 TI - The effects of global change on the threat of exotic arthropods and arthropod borne pathogens to livestock in the United States. AB - Arthropod-borne diseases are an important part of the group of foreign animal diseases that command attention from federal, state, and local animal health authorities in the United States because of the potential for adverse local and regional animal health impacts and also because of possible losses of export markets. Diseases of concern are listed by the US Animal Health Association and also by the Office International des Epizooties. Global change is causing the emergence of newly recognized diseases and altering enzootic and epizootic circumstances for known disease problems. Selected examples of arthropod-borne diseases of importance are discussed in terms of their potential for introduction, spread, and impact on livestock and human health in the United States. PMID- 19120224 TI - The poultry red mite Dermanyssus gallinae as a potential carrier of vector-borne diseases. AB - The poultry red mite Dermanyssus gallinae is an obligatory blood-sucking parasite that is considered to be one of the most important ectoparasites in the poultry industry, mainly because it is responsible for important economic losses, leads to a reduction of welfare of laying hens, and may pose a disease risk to humans. As a result of these problems, much of the current research on this parasite targets new methods of control. Less attention has been paid to the importance of D. gallinae as a carrier of vector-borne diseases. Some authors have mentioned the possible involvement of D. gallinae in the transmission (both in vitro and directly isolated from the mites) of viral and bacterial agents. Our research group has demonstrated the presence of Mycobacterium spp. within D. gallinae. DNA coding for Mycobacterium spp. was successfully amplified from unfed adult D. gallinae, larvae, and eggs by using reverse transcription-polymerase chain reaction targeting the 16S rRNA gene. The results have suggested the possible transovarial and transstadial transmission of pathogens by D. gallinae. PMID- 19120225 TI - Examples of probable host-pathogen co-adaptation/co-evolution in isolated farmed animal populations in the mountainous regions of North Vietnam. AB - In Vietnam, for a number of specific geographical and historical reasons, the mountainous areas have preserved an exceptional diversity of wild and domestic animal species of high socioeconomic interest. This endemic genetic diversity fosters a rapid response to environmental change in mostly isolated local communities and, in particular, fosters the constant adaptation of ecosystems common to humans and farmed and wild animal populations and pathogens. During a 2 year study carried out in several mountainous regions of North Vietnam near the Chinese border, we surveyed 1697 breeders in 249 villages and gathered 5815 biological samples among the four main domesticated species of food animals: chickens, cattle, buffaloes, and goats. Serological analyses were carried out by ELISA on 726 sera in order to assess the prevalence of antibodies specific to two major diseases suspected to be present in the region: avian influenza (AI) and peste des petits ruminants (PPR). The results reported here reveal the presence of antibodies specific to AI, but not the H5N1 highly pathogenic strain, and the presence of antibodies specific to PPR, confirming that this disease, never previously described in Southeast Asia, is present in this region, with no mortality and little or no evidence of clinical cases. These are probably situations of co-evolutive epidemiological equilibrium between pathogen populations, which may have lost their virulence, and animal populations that have acquired genetic resistances over generations, either naturally or through genetic introgression from related wild species better adapted to such pathogens. These results suggest the need for more research, both short-term and, more globally, long-term. PMID- 19120226 TI - Antibiotic clearance of Ehrlichia canis from dogs infected by intravenous inoculation of carrier blood. AB - Ehrlichia canis is the etiologic agent of canine monocytic ehrlichiosis (CME) and is a useful model for zoonotic tick-borne pathogens, many of which infect dogs. The purpose of this study was to evaluate rifampin and doxycycline regimens for clearance of E. canis infections in addition to alleviation of CME. Beagles were infected with E. canis by intravenous inoculation with carrier blood and treated with either rifampin or doxycycline after the acute phase of CME. Improved hematological values demonstrated that both treatments effectively relieved signs of the disease. Peripheral blood from all dogs became PCR negative after antibiotic treatment, suggesting that these infections were eliminated and that rifampin is an effective alternative chemotherapeutic agent for treatment of CME. PMID- 19120227 TI - Frequency and type of renal lesions in dogs naturally infected with leptospira species. AB - The aim of this study was to determine the frequency and type of renal lesions associated with positive titers against Leptospira sp. in a stray dog population. Three hundred fifty pairs of kidneys and an equal number of serum samples were collected from dogs captured by the dog pound of Merida, Yucatan, Mexico. Euthanasia of dogs was performed following the regulations of the Official Mexican Health Ministry (NOM-033-ZOO-1995). Serum samples were evaluated with the microscopic agglutination test, and tissue samples were processed and fixed in paraffin. After staining with hematoxylin and eosin, the frequency of renal lesions was determined and classified. As an additional evaluation, samples with interstitial nephritis were stained by the Warthin-Starry method in order to observe the presence of spirochete forms that could be morphologically compatible with Leptospira spp. We found that 98% of cases presented at least one type of lesion. The main histological lesions found were mesangial proliferative glomerulonephritis (MPGN) in 63.7% (n= 223), mesangial proliferative glomerulonephritis and interstitial nephritis (MPGN+IN) in 34% (n= 119), nephrosclerosis in 0.57% (n= 2), mesangial glomerulonephritis in 0.28% (n= 1), and interstitial nephritis (IN) in 0.28% (n= 1). Thirty-four percent (n= 122) of the dogs were seropositive to Leptospira sp., mainly against serovar canicola. Among dogs with IN (alone or associated with MPGN) (n= 120), 49.1% were seropositive to Leptospira sp., but only 17% of them showed spirochete forms compatible with the bacteria. A statistical association between seropositive dogs and the presence of MPGN+IN was determined (P < 0.0001; odds ratio 2.7, confidence interval 1.7-4.5). We concluded that the frequency of renal lesions found in this study is high and L. canicola is probably the most common circulating serovar in dogs from this area. Dogs that have been in contact with Leptospira spp. have a higher risk of developing renal lesions of the type MPGN+IN. PMID- 19120228 TI - Pharmaceutical proteins in plants. A strategic genetic engineering approach for the production of tuberculosis antigens. AB - Tuberculosis (TB) is a re-emerging disease that is considered a major human health priority as well as an important disease of livestock. TB is also a zoonosis, and Mycobacterium tuberculosis and M. bovis, the human and bovine causative agents, respectively, are very closely related. Protection against TB is essentially achieved through vaccination with the Bacille Calmetle-Guerin (BCG) strain of M. bovis. Protection is, however, incomplete, and novel improved vaccines are currently under investigation. Production of protective antigens in transgenic plants, or "pharming," is a promising emerging approach, and a zoonosis-like TB is a good model for investigating the potential of this approach. Pharma-Planta, a European Commission-funded project and consortium, was set up to address this topic, within which a component is aimed at assessing the production efficacy and stability of the TB antigens in different compartments of the plant cell. This article is meant to introduce this promising approach for veterinary medicine by describing the ongoing project and its specific genetic engineering strategy. PMID- 19120229 TI - Rickettsial agents in Slovakian ticks (Acarina, Ixodidae) and their ability to grow in Vero and L929 cell lines. AB - A total of 80 adult ticks (55 Haemaphysalis inermis, 12 Dermacentor reticulatus, 11 D. marginatus, 2 Ixodes ricinus) were collected from vegetation in three areas of Slovakia (forest and pasture habitat) in central Europe. Forty-six (46 ticks) (57.5%) of all species tested were positive by the hemocyte test, PCR assays based on the gltA and ompA genes showed a Rickettsiaceae infection in 77.5% of the ticks, whereas only one H. inermis tick was positive for Anaplasmataceae on a 16S rRNA-based PCR. Isolation of rickettsiae was attempted on all collected ticks by means of the shell vial technique, 52 isolates of which were inoculated into Vero cells and 28 into L929 cells. Rickettsiae were detected in 50% (40/80) of the cell lines using the Gimenez staining method, whereas 33.8% (27/80) of the cell lines were PCR-positive for Rickettsia species. The presence of rickettsiae was shown by PCR to be around 30.8% (16/52) in Vero and 39.3% (11/28) in L929 cell lines. Sequencing results showed that detected infections were Rickettsia sp., R. raoultii, and Anaplasma phagocytophilum in ticks, and R. slovaca in cell lines. This is the first report of R. raoultii in Slovakia. Observations by electron microscopy of the R. slovaca isolate from Vero cell lines showed a microcapsular layer, typical Gram-negative cell wall, and a cytoplasmic membrane. PMID- 19120230 TI - Toward prevention of cowdriosis. AB - Mass production of Ehrlichia ruminantium variants from different regions of sub Saharan Africa is one of the difficulties that must be overcome in producing a heartwater vaccine. Vaccine productivity can be limited by endogenous induction of interferon (IFN), which inhibits the propagation of Ehrlichia ruminantium (ER) in cell culture. Different kinds of endothelial cells, in which ER multiply efficiently, could be grown in a scalable way in VueLife Teflon bags on Cytodex 3 microcarriers where bead-to-bead transfer of cells occurs. The digital holographic microscope designed at the Universite Libre de Bruxelles allows detection of the most appropriate time to harvest intracellular microorganisms for vaccine production. PMID- 19120231 TI - Prevalence of leptospirosis in dairy cattle from small rural production units in Toluca Valley, State of Mexico. AB - In order to know the seroprevalence of Leptospira spp. in stabled dairy cattle, a study was conducted from 2004 to 2006 in which 416 sera were tested using a microscopic agglutination test conducted on microplates. A collection of culture reference antigens, each representing a serogroup, was used for these tests. Results showed that 10.33% (43) of the animals had antibody titers ranging from 1:100 to 1:1600. The main serovars detected in these tests were L. interrogans serovar hardjo and L. interrogans serovar canicola. It is important to note that these serovars represent a high risk for transmission to other susceptible animal species, between individuals, and to human health. This serological survey provides useful information establishing the presence or absence of these serovars in this type of herd. The range of antigens used in this study included serovars representative of all common serogroups. PMID- 19120232 TI - Milk production increase in a dairy farm under a six-year Brucellosis control program. AB - The present work aims to quantify milk production in a bovine dairy herd during a 6-year brucellosis control program in Hidalgo, Mexico, where bovine brucellosis is endemic. This 6-year longitudinal pilot study comprised 74 monthly samplings to determine the incidence of brucellosis and to quantify daily milk production. To determine the monthly incidence of brucellosis, an average of 346 Holstein cows was examined each month with the card and Rivanol tests. These animals had been vaccinated as calves with the normal dose of Brucella abortus RB51 and were revaccinated annually with a reduced dose. Brucellosis is endemic in Mexico, where the control programs include vaccination and diagnosis; nevertheless, it is uncommon to carry out other essential control practices, such as separation and elimination of positive cows. In this herd, the cows positive to the card and rivanol tests were separated in specific units, especially at the moment of delivery. These cows were placed at the end of the line for milking and were eliminated from the herd at the end of their productive cycle. In this dairy herd, cows were milked three times a day and there was a monthly average of 300 cows in production. At the beginning of this study the prevalence of brucellosis was 8.43%; from days 180-330 the incidence was from 0.51% to 0.90%. Between days 360-570, the incidence diminished to 0%; between days 600 to 1140, it increased to 4.46%; and from days 1440 to 2220 the incidence was kept beneath 1%. The average of dairy milk production for each cow per day started with 24 L, increasing in direct proportion to the decrease in the presence of new cases of brucellosis: in the subsequent years, the production increased successively to 25, 27, 28, 29, and 30 L. The daily average per year of milk production in the herd was also quantified: at the beginning of the study it was 7220 L and in subsequent years was 7470, 7710, 8340, 8790, 8970, and 9150 L, respectively. We conclude that a direct relation existed between the application of a program of brucellosis control and the increase in milk production. PMID- 19120233 TI - Prevalence of pathogens associated with bovine mastitis in bulk tank milk in Mexico. AB - Mastitis as cause of low milk production coupled with the use of medication to control it cause dairy farms to register large economic losses. Microorganisms' prevalence, and somatic cell counts (SCCs) were determined in 112 Holstein bovine herds in two bulk tank milk-screening assays. Detection of Staphylococcus aureus, Streptococcus agalactiae, and Mycoplasma spp. as microorganisms primarily responsible for clinical and subclinical mastitis and their relationship with SCCs was evaluated by Student's t-test and the kappa test. Prevalence of Mycoplasma was 55%; of S. aureus, 30%; of Streptococcus uberis, 37.5%; and of Staphylococcus coagulase-negative, 38.3%. The geometric mean of the SCC was 465,000 cells/mL. No significant differences were observed in the SCCs between the positive and negative samples of pathogens isolated (P > 0.5). There was a low kappa value of Mycoplasma correlation between samplings (kappa value = 0.10). This work aimed to understand the relationship between the prevalence of mastitis pathogens in bulk tank milk and SCCs in bovine herds in the central part of Mexico. PMID- 19120234 TI - Prevention of Blackleg by an immunogen of Clostridium chauvoei. AB - Diseases in livestock caused by Clostridium spp. are of concern in Mexico. There are no good-quality vaccines against these infections, and for this reason several outbreaks have occurred in recent years. The objective of this work was to study the immunogenic capacity of a 156-kDa recombinant protein of Clostridium chauvoei that has shown 80% protection against this disease in guinea pigs. This immunogenic protein was cloned in the expression vector pBluescript and was used to immunize C. chauvoei-free bovine animals that were kept in an endemic area. Three experimental groups were studied. In group 1, 30 bovines were vaccinated by subcutaneous route with one dose of 350 microg/animal of the recombinant protein of 156 kDa. In group 2, 30 bovines were vaccinated with the same concentration of this protein plus aluminium hydroxide as adjuvant. Group 3 was vaccinated with a commercial bacterin by intramuscular route with a dose of 5 mL/animal. In each group, five animals were inoculated with saline solution and remained as controls without vaccination. Blood samples were obtained each month during a 6-month period. Serum samples were analyzed by agglutination test and Western blotting. The recombinant protein of 156 kDa was recognized by serum samples from all the animals in groups 1 and 2. Only two animals from group 3 recognized this protein. During the time of the experiment any cases of this disease were observed. However, other studies with a longer time or greater stress conditions that would favor occurrence of the disease would be required to confirm whether this immunogen is also protective in bovines. PMID- 19120235 TI - Evaluation of BCG vaccine and Mycobacterium bovis culture filtrate proteins against bovine tuberculosis. AB - The objective of this study was to evaluate the degree of protection against TB of three immunogens in calves experimentally challenged with a pathogenic Mycobacterium bovis strain by identification of lesions during necropsy. Twenty four calves free of TB were distributed into four groups: group 1 was inoculated with M. bovis CFPE, group 2 with CFPE/IFN-gamma, group 3 with M. bovis BCG, and group 4 remained as control. After 6 months all animals were challenged with an M. bovis field strain, and 6 months later the animals were euthanized and the presence of lesions was evaluated. Several degrees of protection were observed in the vaccinated groups, particularly in the group vaccinated with BCG, as shown by the absence or decrease in severity of lesions. Vaccination could be useful in high-prevalence zones where it is economically unfeasible to slaughter animals. PMID- 19120236 TI - Human brucellosis in urban and peri-urban areas of Kampala, Uganda. AB - A retrospective case-control study of human brucellosis in urban, peri-urban, and rural areas in Kampala, Uganda was undertaken to find the risks associated with the disease using the medical records of Mulago National Referral Hospital (Mulago Hospital). From the Brucella agglutination test (BAT) records between June 2004 and May 2006, 652 positive results were found. The case-control study showed that living in urban areas was a risk factor for brucellosis. The numbers of improved and cross-breed cattle per 1000 households were calculated on the basis of data obtained from interviews of 75 randomly selected local councils (LCls) in an area between 5 and 20 km radii from the city center of Kampala. Cattle-keeping activities were, however, unpopular in urban areas compared to peri-urban and rural areas. Poor correlation between the distribution of human brucellosis cases and the distribution of cattle suggested that most of the brucellosis cases resulted from consumption of raw milk transported from peri urban and rural areas of Kampala and/or dairy production areas outside Kampala. PMID- 19120237 TI - Development of specific oligonucleotide probes to detect Vibrio species. AB - Many species of Vibrio are responsible for diseases in marine organisms and for economic losses to the aquaculture industry. The aim of this preliminary study was to obtain species-specific DNA zones to be used as potential probes from a phylogenetic analysis of the 23S ribosomal RNA (rRNA) gene of different Vibrio species from marine and human organisms. Species-specific probes were identified for V. parahaemolyticus, V. fortis, V. splendidus, and for two clusters of taxonomically related species, namely V. harveyi/campbelli and V. lentus/aestuarianus. A reverse line blot assay showed that the designed probes can specifically detect the different Vibrio species, thereby proving that these probes can be used to evaluate the presence of pathogenic and nonpathogenic Vibrio species in the sea and in marine organisms to assist in the investigation of environmental risks. PMID- 19120238 TI - Burden of zoonotic diseases in Venezuela during 2004 and 2005. AB - Emerging zoonotic diseases have increased in importance in human and animal health during the last 10 years. Each of these diseases has emerged from an unsuspected quarter and has caused severe problems. In countries, such as Venezuela, the burden of these zoonoses has been understudied. In this report we analyze the status of such diseases in Venezuela for the period 2004-2005. During this period a significant number of animal and human cases of these diseases have been found, particularly for rabies, encephalitis, leishmaniasis, and cysticercosis among others. Surveillance needs to be reinforced for these zoonoses. PMID- 19120239 TI - Viral association with the elusive rickettsia of viper plague from Ghana, West Africa. AB - We previously reported a rickettsial heartwater-like disease in vipers from Ghana that resembled heartwater in its gross lesions, was apparently transmitted by ticks (Aponomma and Amblyomma), and responded clinically favorably to early treatment with tetracycline. Cell culture showed consistent cytopathic effects in bovine endothelial cells, viper cells, and mouse cells, and inhibition of cytopathic effect by tetracycline in vitro. A type D retrovirus was observed in vacuoles in all infected cells. The virus and rickettsia infection was associated with transfer of cytopathic effect, regardless of cell species. Close association of virus and rickettsia may indicate a dual infection etiology of viper plague. PMID- 19120240 TI - Mammal and bird species distribution at the woodland-pasture interface in relation to the circulation of ticks and pathogens. AB - The tick vector Ixodes ricinus can be found in pastures in landscapes where pasture and woodland intermix, such as in the bocage. To identify which mammal or bird species could potentially transport ticks between woodland and pasture, we did a species inventory on four French bocage sites in 2005. Fifty-eight species were observed, 21 of which were found in three different habitats: the middle of the pasture, the hedgerow ecotone, and the woodland. Because the bocage is a meeting point for wildlife, domestic animals, and humans, species using the three habitats could potentially be epidemiologic agents for circulation of tick vectors as well as of pathogens. Thus, the bocage could be used as a model landscape for the study of epidemiologic cycles and potential emergence of multiple-host pathogens. PMID- 19120241 TI - Zoonoses and climate variability. AB - Leishmaniasis in the Americas is transmitted by Lutzomyia spp., which have many animal reservoirs. Previous studies indicated potential changes in vectors of climate-related distribution, but impact outcomes need to be further studied. We report climatic and El Nino events during 1985-2002 that may have had an impact on leishmaniasis in 11 southern departments of Colombia: Amazonas, Caqueta, Cauca (Ca), Huila, Meta (Mt), Narino, Putumayo (Py), Tolima, Valle (Va), Vaupes (Vp), and Vichada. Climatic data were obtained by satellite and epidemiologic data were obtained from the Health Ministry. NOAA climatic classification and SOI/ONI indexes were used as indicators of global climate variability. Yearly variation comparisons and median trend deviations were made for disease incidence and climatic variability. During this period there was considerable climatic variability, with a strong El Nino for 6 years and a strong La Nina for 8. During this period, 19,212 cases of leishmaniasis were registered, for a mean of 4756.83 cases/year. Disease in the whole region increased (mean of 4.98%) during the El Nino years in comparison to the La Nina years, but there were differences between departments with increases during El Nino (Mt 6.95%, Vp 4.84%), but the rest showed an increase during La Nina (1.61%-64.41%). Differences were significant in Va (P= 0.0092), Py (P= 0.0001), Ca (P= 0.0313), and for the whole region (P= 0.0023), but not in the rest of the departments. The importance of climate change is shown by shifts in insect and animal distributions. These data reflect the importance of climate on transmission of leishmaniasis and open further investigations related to forecasting and monitoring systems, where understanding the relationship between zoonoses and climate variability could help to improve the management of these emerging and reemerging diseases. PMID- 19120242 TI - Use of cymelarsan in goats chronically infected with Trypanosoma evansi. AB - Toxicity and therapeutic trials using Cymelarsan (an arsenical compound) against Trypanosoma evansi infection were carried out using chronically infected goats. For the toxicity trial, 40 goats were divided into four groups of 10 animals each; the first three groups received s.c. injections of 5, 10, and 15 mg/kg bw of Cymelarsan, respectively, and the last one served as control. No systemic reaction was observed in any goat throughout the experiment. For the therapeutic trial, 15 adult female goats were inoculated intravenously with at least 1 x 10(5)T. evansi isolated in the Canary Islands. Six months after inoculation, the animals were treated with Cymelarsan at single dose of 0.3 mg/kg (5 animals), 0.5 mg/kg (5 animals), and 0.625 mg/kg (5 animals). At 4 and 6 weeks after treatment, two goats belonging to 0.3 mg/kg group showed recurrence of trypanosomes. Parasitemia, however, was negative in all animals belonging to 0.5 and 0.625 mg/kg groups until the end of the experiment (6 months after treatment). Thus, it can be concluded that Cymelarsan is a safe trypanocidal drug for goats and that the curative dose is 0.5 mg/kg or above. PMID- 19120243 TI - Fluorescence-based detection of Leishmania infantum DNA in phlebotomus vectors. AB - The geographical diffusion of leishmaniasis can depend on the factors limiting the distribution of sandfly vectors. In Sicily, as in all Mediterranean areas, sandflies are present almost all year round because the climate permits perpetuation of this vector's life cycle. Transmission can occurs in rural and domestic habitats through the bite of infected female phlebotomine sandflies. In Italy, the visceral form of leishmaniasis is commonly found, which is due exclusively to L. infantum. Two species of sandfly are considered the main vectors: Phlebotomus perniciosus and P. perfiliewi. Fluorescence resonance energy transfer technology was used to determine the parasite load in phlebotomine vectors, and the test was targeted on a 117-bp fragment of kinetoplast DNA minicircles. The assay was evaluated, focusing on analytical sensitivity, discriminatory power, and reliability of quantification. During 2005, a total of 1686 sandflies were trapped in various Sicilian provinces and in farms randomly selected using black light traps. We found 20, 30, and 16 sandflies positive for Leishmania for each kind of analyzed phlebotomine sandfly, respectively, corresponding to 6.5% for the gravid, 2.7% for the fed, and 6.3% for other groups. Previously the insects were identified on the basis of morphology and the most prevalent sandfly species were P. Sergentomyia minuta, P. perfiliewi, and P. perniciosus. PMID- 19120244 TI - Secretome of animal trypanosomes. AB - Animal trypanosomosis is one of the most severe constraints to agricultural development in sub-Saharan Africa and is also an important disease of livestock in Latin America and Asia. The causative agents are various species of protozoan parasites belonging to the genus Trypanosoma, among which T. congolense and T. evansi are the major pathogenic species. The extracellular position of trypanosomes obliges us to consider both the parasite and its excreted/secreted factors in the course of the physiopathologic process. The advent of proteomics led us to propose a comparative approach of the proteome (i.e., the whole parasite content) and the secretome (i.e., naturally excreted/secreted molecules) of T. congolense and T. evansi with particular attention to common and specific molecules between strains of differing virulence and pathogenicity. The molecular identification of differentially expressed trypanosome molecules correlated with either the virulence process or the pathogenicity will provide new potential molecular targets for improved field diagnosis and chemotherapy of animal trypanosomosis. PMID- 19120245 TI - Therapeutic DNA vaccine against Trypanosoma cruzi infection in dogs. AB - Chagas' disease is an important health problem in most Latin American countries, and a concern in dog populations, which act as a reservoir. We showed in previous studies that a therapeutic DNA vaccine could partially control the pathology after Trypanosoma cruzi infection in mice, and this vaccine may represent an alternative treatment for Chagas' disease. Here we evaluated the therapeutic efficacy of this vaccine in experimentally infected dogs for up to 2 months after infection. Our results suggest that DNA vaccine treatment may affect the immune response and delay Chagas' disease progression in T. cruzi-infected dogs, and confirm the potential of this novel treatment. PMID- 19120246 TI - Characterization of DNA sequences that confer complement resistance in Leishmania chagasi. AB - Serial passage of axenically cultured Leishmania chagasi promastigotes results in a progressive diminution in resistance to complement-mediated lysis (CML), whereas high CML resistance is seen in infectious metacyclic promastigotes from the sandfly vector as well as metacyclic-like promastigotes within low-passage cultures at stationary growth phase. As we previously reported, in a screen seeking to identify novel genes involved in CML resistance: (1) a genomic cosmid library derived from DNA of CML-resistant L. chagasi promastigotes was transfected into high-passage (constitutively CML-sensitive) L. chagasi promastigotes; (2) transformants were screened for acquisition of CML-resistance; (3) multiple cosmid-transfectants exhibited partial CML resistance; and (4) the sequence for one of the cosmids (Cosmid 51) was determined. This report extends the analysis of Cosmid 51, and identifies by deletion analysis a subregion of the cosmid insert that is critical to the CML-resistance phenotype of Cosmid 51 transformants. We also report the sequence determination and initial CML resistance activity of another cosmid that also confers partial resistance to CML. PMID- 19120247 TI - Presence of Trypanosoma theileri in Spanish Cattle. AB - Trypanosoma theileri (Laveran, 1902) has been diagnosed in many countries and is commonly considered as a nonpathogenic hemoparasite, although some authors have described clinical signs in cattle infected with T. theileri. In April and May, 2005, 12 blood samples were received at the Exopol Diagnostic Laboratory (Zaragoza, Spain) from a Spanish bull-fighting farm located at Seville province. Clinical exploration of the animals revealed fever, progressive weight loss, anemia, and frequent recumbent position. Microscopic examination showed Theileria spp. in all cases (12), and in four of them, T. theileri was also observed. The clinical picture observed in the animals could be compatible with T. theileria infection. However, the contribution of T. theileri to the clinical signs seen at least in four cases is unknown. Further studies are necessary to determine the pathogenicity of T. theileri in the different animal species. To our knowledge, this is the first isolation of T. theileri in Spain. PMID- 19120248 TI - Use of phlebotomine sand flies as indicator of leishmania prevalence in an endemic area. AB - An entomological survey was conducted on phlebotomine sand flies to discover the presence of Leishmania parasites in six Sicilian provinces. During the season from June to October 2005, 20,346 sand fly specimens were collected by using a "black light" trap in eight cattle and sheep farms. The most prevalent species was Phlebotomus perfiliewi (51.60%) followed by P. perniciosus (24.37%), and Sergentomyia minuta (24.03%). Among the 11,441 females collected, 284 (2.70%) were gravid and 708 (6.74%) were blood-fed females. A total of 194 gravid females, 274 blood-fed females, and 254 females with no evidence of blood meal were analyzed for L. infantum by PCR analysis, showing 9.79% gravid females, 5.84% blood-fed females, and 5.51% females with no evidence of blood meal positive for Leishmania DNA. These results support the use of gravid female sand flies for the evaluation of prevalence of Leishmania. PMID- 19120249 TI - Leishmania DNA quantification by real-time PCR in naturally infected dogs treated with miltefosine. AB - A new drug that has just become available in India for treatment of human visceral leishmaniasis (VL) is miltefosine, an alkyphospholipid that was originally developed as an oral antineoplastic agent. Miltefosine is not only directly toxic for Leishmania parasite, but it also enhances both T cell and macrophage activation and production of microbicidal reactive nitrogen and oxygen intermediates. It is highly effective in the treatment of Leishmania infection in mice and human beings. However, adverse effects in dogs treated with miltefosine have been reported, but there are no data on the efficacy of this drug for the treatment of canine visceral leishmaniasis (CVL). The aim of this study was to use a real-time PCR assay to monitor the Leishmania load in the blood samples and lymph node aspirates of 18 naturally infected dogs before and after treatment with miltefosine (2 mg/kg for 30 days). The results of our study showed that the therapy with miltefosine shows a drastic and progressive reduction of parasite load in lymph node aspirates, but does not suppress the parasite in lymph nodes. In all dogs the real-time PCR assay demonstrated an irregular presence of parasites in blood. Therefore, blood does not seem a suitable substrate for the purpose of quantifying Leishmania DNA. PMID- 19120250 TI - Serologic and molecular evaluation of Leishmania infantum in cats from Central Spain. AB - Infection by different Leishmania spp. in cats has been reported in many countries. In Spain, since the first Leishmania infection described in 1933, sporadic clinical cases in cats have been reported. Various serologic studies performed in other areas of Spain have shown seroprevalences ranging between 1.7 and 60%. The aim of the present study was to determine the prevalence of leishmaniasis in cats from Central Spain (Madrid), and to assess the existence of associations between Leishmania infantum infection and relevant data obtained from each cat. Two-hundred thirty-three cats attended at the Veterinary Teaching Hospital in Madrid between September 2005 and June 2006 were tested for L. infantum using the indirect immunofluorescent antibody (IFA) test (cutoff: 1:100) and PCR. PCR testing was performed on the samples to detect Leishmania infection, targeting the kinetoplast DNA (kDNA). Our results showed a seroprevalence of 1.29% (3/233) using IFA test. Another seven cats were also seroreactive to L. infantum one dilution under the cutoff (1:50). Considering all the seroreactive samples, the percentage of positive animals to L. infantum was 4.29%. Only one of the cats (0.43%) included in the study was PCR-positive. Relative lymphocytosis and an increase in alanine aminotransferase (ALT) value were statistically associated with seroreactivity to L. infantum. Our results demonstrate the presence of cats seroreactive to L. infantum in Central Spain, an endemic area for this disease in dogs. PMID- 19120251 TI - Comparing therapeutic efficacy between ivermectin, selamectin, and moxidectin in canaries during natural infection with Dermanyssus gallinae. AB - The aim of this study was to compare the efficacy of three spot-on drugs on canaries during Dermanyssus gallinae natural infections and during the breeding season. Three groups of canary couples (seven couples each) were included: group A was treated with ivermectin, B with selamectin, and C with moxidectin. All the drugs were administered topically infrascapularly. The parasitic charge was estimated before the treatment (t(0)) and after 8 (t(1)), 16 (t(2)), 24 (t(3)), and 32 (t(4)) days following the initial treatment. No significant differences were detected among the three tested drugs for the five repeats for each of the four mite stages (egg, larva, nymphs, and fed and unfed adults). With regard to the decrease in the mean numbers of red mites, ivermectin and selamectin exerted their efficacy at t(2), contrary to moxidectin at t(3). PMID- 19120252 TI - Diseases of backyard turkeys in the Mexican tropics. AB - With the purpose of identifying the causes of disease and death in backyard turkeys, 768 interviews with growers were carried out in communities of the coastal region of Oaxaca, Mexico. Later, a clinical sanitary evaluation was performed on five randomly selected turkeys of different age and sex. The laboratory tests included: necropsy for clinical diagnosis of diseases and for a histopathological examination of tissues with macroscopic lesions to identify microscopic lesions; hemagglutination inhibition tests to diagnose Newcastle disease and avian influenza; agglutination test to identify Mycoplasma gallisepticum, M. synoviae, and Salmonella pullorum; and enzyme-linked immunosorbent assay to identify infectious bronchitis and infectious bursal disease. Both internal and external parasites were identified. The data obtained by the interviews showed that sudden death, avian pox, respiratory disease, and diarrhea were the most important manifestations in backyard turkeys. The laboratory tests found avian pox, avian influenza, and infectious bronchitis; sensitivity to M. synoviae and M. gallisepticum; protozoal infections by Eimeria meleagridis and E. dispersa; internal parasite infestation by Heterakis gallinae and Ascaridia gallinae; and mycotoxicoses. PMID- 19120253 TI - Epidemiological scenario of giardiosis in dogs from central Italy. AB - In order to update and implement the data on the epidemiological situation of giardiosis in the Italian arena, stool samples from 240 dogs living in the Abruzzo region (central Italy) were examined for prevalence, risk factors, and genotypes. Giardia duodenalis cysts were detected in 26.6% of the dogs and kennel dogs tested. Dogs younger than 12 months (38%) and with diarrhea (46%) were statistically more affected. Species-specific G. duodenalis assemblages (C and D) were identified in kennel and in privately owned dogs, while the zoonotic assemblage A was identified in privately owned dogs. In light of these results, giardiosis in dogs, and mainly in well-cared dogs, is still a problem of concern and may pose a public health risk. PMID- 19120254 TI - Localization of Entamoeba histolytica amebopore in amebic liver abscesses in hamsters. AB - Amebopore was purified from axenically grown trophozoites of the Entamoeba histolytica strain HM1:IMSS. The purification procedure involved Mono Q anion exchange chromatography and electroelution. Sequence analysis of the final product revealed that amebopore A was completely pure. Polyclonal antibodies against the purified amebopore were obtained from rabbits, and Western blot studies demonstrated their specificity. Sections of experimental, acute (1, 2, 3, and 4 days), amebic liver abscesses produced in hamsters were stained with the anti-amebopore antibody; in all the analyzed stages, amebopore appeared as a constitutively expressed cytoplasmic molecule in trophozoites. No extracellular or hepatocyte-membrane amebopore was found. This study is the first to trace amebopore in an in vivo model of amebic liver abscesses. PMID- 19120255 TI - Secreted proteins of Avibacterium paragallinarum are lethal for chicken embryo. AB - Avibacterium paragallinarum causes infectious coryza in chickens. This bacterium secretes proteins of 110 kDa (a putative RTX protein) and 120 kDa. Expression of these proteins increases by the addition of CaCl(2), MgSO(4), MnSO(4), or ferric ammonium citrate and diminishes with CuSO(4) or ZnCl(2). Protein expression is optimal at 37 degrees C and pH 7.5. Mortality (90-100%) of chicken embryos was observed when secreted proteins (SPs) from A. paragallinarum reference or field isolates (serogroup A or C) were inoculated via yolk sac and was not observed when SPs from A. avium, a chicken respiratory tract indigenous bacterium, were inoculated. A. paragallinarum SPs could contain toxins responsible for the embryo deaths. Indeed, presence of the putative RTX protein of 110 kDa was confirmed by Western blotting with antibodies against the Actinobacillus pleuropneumoniae RTX ApxI, a closely related RTX protein. PMID- 19120256 TI - Virulence of Metarhizium anisopliae to eggs and immature stages of Stomoxys calcitrans. AB - Stomoxys calcitrans control is primarily based on the use of chemical products; however, new methods for control have been investigated. Biological control has been proposed as a promising alternative to traditional chemicals. The current study evaluated three methods in vitro to treat S. calcitrans eggs with the fungus Metarhizium anisopliae and assessed the virulence of this fungus to immature stages of this fly. In the first method of egg exposure, eggs were immersed in conidial suspensions and transferred to Petri plates with high humidity. A high mortality was observed in both treated and control groups. In the second method, the eggs were transferred to rearing medium right after immersion in conidial suspensions; the suspensions with highest conidial concentrations, 10(7) and 10(8) conidia mL(-1), caused 96.25% and 100% mortality, respectively. In the third method, the eggs were sprayed with conidial suspensions on the rearing medium. A high mortality was observed at 10(7) and 10(8) conidia mL(-1), 78.3% and 100%, respectively. The second method had the lowest mortality in the control group and was considered the most useful method for evaluating the effect of entomopathogenic fungi on S. calcitrans eggs. While eggs were very susceptible to M. anisopliae infection, larvae and pupae were not. PMID- 19120257 TI - Virulence of Metarhizium anisopliae and Beauveria bassiana to Ctenocephalides felis felis. AB - Ctenocephalides felis felis is a flea species that parasitizes dogs and cats. The current study verified the in vitro virulence of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana to eggs and adults of C. felis felis. The fungal isolates were cultivated on rice grains in plastic bags. Conidial suspensions were then prepared by immersing the grains in Tween 80 aqueous solution (0.01%). Eggs and adults of C. felis felis were obtained from a laboratory colony. Experiments were composed of one control and four treatment groups (10(5), 10(6), 10(7), and 10(8) conidia mL(-1)). Both insect pathogens inhibited egg hatchability and caused adult mortality. Among isolates used in this study, M. anisopliae (isolate E9) was the most successful at inhibiting hatching, while B. bassiana (isolate 986) was the most powerful for causing mortality in adults. The results indicate that these entomopathogenic fungi have promising potential for use in controlling both C. felis felis egg and adult stages. PMID- 19120258 TI - Toward a PCR-independent molecular diagnosis of veterinary and medically relevant pathogenic organisms. AB - Bloodstream infections caused by bacteria and fungi are a major problem worldwide. These bloodstream infections can affect both people and livestock, placing a significant burden upon developed and developing economies. In this paper we describe a multiplexed testing format, which can identify a range of bacteria and fungi within a single blood sample. Key to this technique is the specificity and sensitivity of the nucleotide probes that capture the sample. The sensitivity and specificity of the probes may allow detection of disease-causing microorganisms without the need for polymerase chain reaction amplification if the dynamics of probe binding can be observed in real time. PMID- 19120259 TI - Hematological and plasma biochemistry of the adult wild African grasscutter (Thryonomys swinderianus). AB - Hematological and plasma biochemical values of wild grasscutters were evaluated to determine their potential to transmit zoonotic pathogens. Three 5-mL blood samples were collected from each of 1000 grasscutters caught in the wild for hematology, biochemical, and parasitological tests. Hematological and biochemical values were compared with those from captive-reared grasscutters. There are significantly (P < 0.05) higher lymphocyte, eosinophil, and basophil values for wild grasscutters compared to those that are captive reared. Parasitological examination revealed a 15% prevalence of blood protozoa in the wild grasscutters. Blood pathogens encountered were Trypanosoma sp. (66.7%) and Plasmodium sp. (33.3%), with 20.7% mixed infection. Sex does not significantly (P > 0.05) affect blood protozoa infection, while season does. We therefore concluded that wild grasscutters serve as efficient reservoir hosts for agents of African trypanosomiasis and malaria in the tropical humid rainforest region of Nigeria. PMID- 19120260 TI - The IDS conducts standardization of assays for the measurement of autoantibodies and of T cells in autoimmune diabetes. Preface. PMID- 19120262 TI - Type 1 Diabetes TrialNet--an international collaborative clinical trials network. AB - Type 1 Diabetes TrialNet is an international consortium of clinical research centers aimed at the prevention or delay of type 1 diabetes (T1D). The fundamental goal of TrialNet is to counter the T1D disease process by immune modulation and/or enhancement of beta cell proliferation and regeneration. To achieve this goal, TrialNet researchers are working to better understand the natural history of the disease, to identify persons at risk, and to clinically evaluate novel therapies that balance potential risks and benefits. The particular focus is on studies of preventive measures. In addition, TrialNet evaluates therapies in individuals with newly diagnosed T1D with preserved beta cell function to help determine the risk/benefit profile and gain an initial assessment of potential efficacy in preservation of beta cell function, so that promising agents can be studied in prevention trials. In addition, TrialNet evaluates methodologies that enhance the conduct of its clinical trials, which includes tests of outcome assessment methodology, the evaluation of surrogate markers, and mechanistic studies laying the foundation for future clinical trials. PMID- 19120263 TI - What will it take to get therapies approved for type 1 diabetes? AB - The development of therapies for T1D has been neglected in favor of efforts in advancing therapies for the larger T2D population. Pharmaceutical companies have also been deterred by lack of clarity around the regulatory expectations for such therapies. The prospects for therapy for new-onset T1D have brightened in some respects because of convergence among regulators and clinical experts in views about how these therapies should be assessed. The most important consensus is that the primary efficacy end point for treatments directed at the underlying autoimmune cause of T1D should be endogenous insulin secretion, as reflected by standardized C-peptide measurements. Most T1D therapeutic development efforts are directed at new-onset disease, which represents a small proportion of the entire T1D population. A major deficiency in T1D therapeutic development is the lack of activity in advancing therapies for people with established T1D, a population that far outnumbers those with new-onset disease. Complete remission of new-onset or established T1D will almost certainly require a combination of two or more therapies to address the underlying cause of the disease and restore normal insulin function. PMID- 19120264 TI - Islet glia, neurons, and beta cells. AB - Type 1 diabetes (T1D) is caused by autoimmune beta cell destruction. The early events triggering T1D and the forces that keep diabetic autoimmunity pancreas specific have been unclear. Our discovery that autoimmune islet destruction is not beta-cell-exclusive but includes cytotoxic T cell targeting of peri-islet glia, evoked the possibility that T1D pathogenesis may involve neuronal elements beyond beta cell/immune interactions. Recently, we have found that sensory afferent neurons are a critical component in prediabetes initiation, promoting islet inflammation through altered glucose homeostasis and progressive beta cell stress. These factors orchestrate a catastrophic cascade culminating in insulin insufficiency mediated by an autoimmune-prone host. This neuro-immuno endocrinological triad explains diabetic inflammation as a consequence of local neuropeptide deficiency, leading to an innovative concept of disease pathogenesis with novel therapeutic implications. PMID- 19120265 TI - Hedgehog signaling during expansion of human pancreatic islet-derived precursors. AB - A detailed understanding of the molecular process involved in the proliferation of pancreatic precursor cells would provide key elements for developing new therapeutic strategies to cure type 1 diabetes. In the present study we investigated the potential involvement of hedgehog signaling in proliferating human pancreatic islet-derived mesenchymal (hPIDM) cells, a population of cells that can be successfully expanded and induced to differentiate into an insulin secreting phenotype. Here we report that in these precursor cells a hedgehog signaling pathway is activated, as shown by Gli1 expression, and that a dose dependent inhibition of such a pathway by cyclopamine results in a significant reduction of cell proliferation. PMID- 19120261 TI - The Environmental Determinants of Diabetes in the Young (TEDDY) Study. AB - The etiology of type 1 diabetes (T1D) remains unknown, but a growing body of evidence points to infectious agents and/or components of early childhood diet. The National Institutes of Health has established the TEDDY Study consortium of six clinical centers in the United States and Europe and a data coordinating center to identify environmental factors predisposing to, or protective against, islet autoimmunity and T1D. From 2004-2009, TEDDY will screen more than 360,000 newborns from both the general population and families already affected by T1D to identify an estimated 17,804 children with high-risk HLA-DR,DQ genotypes. Of those, 7,801 (788 first-degree relatives and 7,013 newborns with no family history of T1D) will be enrolled in prospective follow-up beginning before the age of 4.5 months. As of May 2008, TEDDY has screened more than 250,000 newborns and enrolled nearly 5,000 infants--approximately 70% of the final cohort. Participants are seen every 3 months up to 4 years of age, with subsequent visits every 6 months until the subject is 15 years of age. Blood samples are collected at each visit for detection of candidate infectious agents and nutritional biomarkers; monthly stool samples are collected for infectious agents. These samples are saved in a central repository. Primary endpoints include (1) appearance of one or more islet autoantibodies (to insulin, GAD65 or IA-2) confirmed at two consecutive visits; (2) development of T1D. By age 15, an estimated 800 children will develop islet autoimmunity and 400 will progress to T1D; 67 and 27 children have already reached these endpoints. PMID- 19120267 TI - Developing a novel model system to target insulin-reactive CD8 T cells. AB - CD8 T cells play an important role in autoimmune diabetes development, and therefore removing these cells may protect against disease. To test this, we designed a novel method using engineered cells (InsCD3-zeta) to target insulin specific CD8 T cells. Insulin-reactive target cells were cultured with InsCD3 zeta CD8 T cells and cytotoxicity was assessed. Activated, but not naive, InsCD3 zeta CD8 T cells readily killed insulin-reactive target CD8 T cells. This approach to immunotarget relevant pathogenic CD8 T cells may be a therapeutic option to delay or prevent type 1 diabetes. PMID- 19120266 TI - Humanized mice for the study of type 1 diabetes and beta cell function. AB - Our understanding of the basic biology of diabetes has been guided by observations made using animal models, particularly rodents. However, humans are not mice, and outcomes predicted by murine studies are not always representative of actual outcomes in the clinic. In particular, investigators studying diabetes have relied heavily on mouse and rat models of autoimmune type 1-like diabetes, and experimental results using these models have not been representative of many of the clinical trials in type 1 diabetes. In this article, we describe the availability of new models of humanized mice for the study of three areas of diabetes. These include the use of humanized mice for the study of (1) human islet stem and progenitor cells, (2) human islet allograft rejection, and (3) human immunity and autoimmunity. These humanized mouse models provide an important preclinical bridge between in vitro studies and rodent models and the translation of discoveries in these model systems to the clinic. PMID- 19120268 TI - Lowered expressions of the NF-kappaB family members in dendritic cells from NOD mice are associated with a reduced expression of GATA-2. AB - In type 1 diabetes, dendritic cells (DCs) display defective phenotype and function and possibly play crucial roles in the pathogenesis of this disease. In the present study, we compared transcription profiles of CD11c(+) bone marrow (BM)-derived DCs from NOD mice with those from NON mice, focusing on the NF kappaB/Rel family members and associated molecules. The BMDCs from NOD mice displayed reduced mRNA expressions of NF-kappaB components, p65, p50, p52, and RelB, compared to NON mice: the proportions of each molecule relative to those of NON DCs were 53.9, 54.1, 54.0, and 37.0%, respectively, which were accompanied with lowered expressions of downstream immunomodulatory molecules, including IL 6, CD80, CD86, 4-1BB, and CD40. The reduction of NF-kappaB components possibly underlies the defective phenotype and function of DCs from NOD mice, and could predispose to autoimmunity. PMID- 19120269 TI - Measurement of CD8 T cell responses in human type 1 diabetes. AB - Type 1 diabetes (T1D) is a T cell-mediated autoimmune disease targeting pancreatic beta-cells. Despite this textbook definition, it is quite striking that neither the diagnosis nor the therapy nor the follow-up of T1D "belong" to immunologists, but rather to endocrinologists whose only option is to limit the consequences of the disease. Immune therapies would seem better suited to correct the causes of T1D, but critical laboratory tools are missing for early diagnosis, prognostic stratification, and therapeutic follow-up. The immune markers routinely available are limited to autoantibodies, which have some intrinsic limitations. Because T cells are central pathogenic actors of T1D, the quest for their measurement appeared to offer a path towards new autoimmune markers. Given the strong association between T1D susceptibility and the HLA class II locus, investigators have long been focused on CD4(+) T cells. However, data gathered in the NOD mouse and the examination of human insulitis point to a critical role of CD8(+) T cells in the pathogenesis of T1D. These observations have revived interest in trying to measure CD8(+) T cell responses in human T1D. Achievement of this goal mainly depends on two factors. First, the relevant epitopes need to be identified. Second, appropriate readouts and measurement techniques need to be selected. This review summarizes recent advances on both of these battlefronts, and discusses the potential clinical applications of T cell assays. PMID- 19120270 TI - Comparative analysis of the islet proteome between NOD/Lt and ALR/Lt mice. AB - Although alloxan-resistant (ALR) mice share 70% of the NOD genome, ALR islets are remarkably resistant to oxidative stress and autoimmunity. Using 2D PAGE comparative analysis, we have characterized 55 proteins that are differentially expressed between the NOD and ALR islet proteome. Ten proteins were found to be highly expressed in the NOD islets. Typically, proteins associated with pancreatic inflammation and autoimmunity, such as amylase and carboxypeptidase, are highly expressed in the NOD islets. Forty-five proteins showed significantly higher expression in the ALR islets. Among these, 30 are proteins implicated in the regulation of intracellular stress including heat-shock proteins, disulfide isomerase-associated proteins, ROS detoxification enzymes, and apoptotic regulators. Our results clearly demonstrate that the 30% unique ALR genome encodes protective determinants expressed at islet levels, which render the islets of this strain of mice resistant to oxidative stress and autoimmunity. PMID- 19120271 TI - MicroRNA genes. AB - Human type 1 diabetes (T1D) is a chronic autoimmune disorder with complex genetic inheritance. To date, more than 19 insulin-dependent diabetes mellitus (IDDM) susceptibility loci have been mapped to specific chromosome regions in the human. MicroRNAs (miRNAs) are a recently discovered class of evolutionarily conserved small noncoding RNAs that negatively regulate the expression of protein-coding genes without affecting mRNA levels. There are a growing number of reports that miRNAs link to the regulation of different biological pathways associated with human diseases. However, the potential role of miRNAs in human T1D is still unknown. To investigate the possible involvements of miRNAs in human T1D on a genome-wide basis, we have mapped 530 miRNAs and compared their locations to the current IDDM loci. We found that at least 27 miRNAs are located in 9 human IDDM loci. More interestingly, some of them potentially target autoimmune- and beta cell-related genes. Our data represent a genome-wide search for a potential correlation between the genomic position of miRNAs and specific IDDM loci, indicating that miRNAs may be susceptibility candidates or biomarkers for human T1D. PMID- 19120272 TI - Independent association of tumor necrosis factor polymorphism with type 1 diabetes susceptibility. AB - The contribution of SNPs in TNF genes to type 1 diabetes (T1D) is not well established and may be confounded by the linkage disequilibrium within the HLA genes. Seven SNPs in the TNF genes (TNFA and TNFB) were genotyped in a Korean cohort (398 T1D patients and 1422 nondiabetic controls), along with HLA DRB1, DQB1, and MICA (MHC class I chain-related genes). Among them, three SNPs (TNFB+318, TNFA-857, and TNFA-308) and two common TNF haplotypes showed significant association with the risk of T1D (P= 5 x 10(-3)-10(-5)). T1D patients were more often heterozygous for the alleles at the TNFB+318 (OR = 1.7, P= 10( 3)) and TNFA-308 (OR = 1.7, P < 10(-5)) than were the controls. Genetic association analyses of the DRB1, DQB1, and MICA alleles with the risk of T1D revealed dramatic associations in several alleles as expected. Independent analyses to discern the genetic effects of TNF polymorphisms on the risk of T1D suggested that these genetic influences might be not totally dependent on the nearby HLA genes. Our results support the hypothesis that two susceptibility loci in the MHC (one in the HLA class II and another in the central MHC region) act epistatically to increase susceptibility to T1D. PMID- 19120273 TI - HLA class I supertypes in type 1 diabetic children in an urban children's hospital. AB - Type 1 diabetes is an autoimmune disorder characterized by progressive destruction of insulin-secreting beta cells of the pancreas, in which CD8(+) T cells play a critical role. The diversity in the HLA alleles expressed among various racial and ethnic groups leads to great variability in antigen presentation and recognition by CD8(+) T cells in the context of MHC class I molecules. To date, studies aimed at identifying disease-relevant antigenic epitopes have focused on using mice transgenic for HLA-A*0201, a common allele, particularly among Caucasians. We present HLA class I typing data from 88 children with type 1 diabetes at the Children's Hospital at Montefiore, where the patient population is ethnically diverse, but largely minority. When categorized into the HLA supertypes A2, A3, B7, and C1, 77% of those studied have alleles belonging to at least one supertype, and of these patients, 65% do not belong to the A2 supertype, which is the supertype represented by the HLA-A*0201 allele. These results support the need for studies using HLA transgenic mice expressing MHC molecules representative of a variety of HLA supertypes, particularly when searching for antigenic epitopes applicable for study among largely urban, minority pediatric populations. PMID- 19120274 TI - Congenic mapping of the MHC-linked susceptibility to type 1 diabetes in the NOD mouse: at least two genes contribute to the Idd16 effect. AB - Major histocompatibility complex (MHC)-linked susceptibility to type 1 diabetes consists of multiple components. Previous studies with NOD mice congenic for the MHC from a sister strain, the CTS mouse, mapped Idd16, a second component of MHC linked susceptibility to type 1 diabetes, in the region adjacent to, but distinct from Idd1 in the class II region. In this study, three lines of subcongenic strains were established from the original congenic strain. Phenotypic analysis of the strains indicated that MHC-linked susceptibility to type 1 diabetes consists of at least three components: Idd1, Idd16.2 adjacent to Idd1, and Idd16.1 telomeric to Idd16.2. PMID- 19120275 TI - STAT4 polymorphism is associated with early-onset type 1 diabetes, but not with late-onset type 1 diabetes. AB - In an effort to discover non-HLA genes affecting susceptibility to type 1 diabetes (T1D), we have investigated the association of polymorphisms in STAT4, an important signaling molecule of IL-12, gammaIFN, and IL-23, in a sample of 389 T1D patients and 152 nondiabetic controls in Korea. Four SNPs on chromosome 2q, which were recently found to be associated with rheumatoid arthritis, were examined for association and linkage disequilibrium. We found that neither alleles or genotypes among all four SNPs nor reconstructed haplotypes of the three SNPs within the same LD block (rs7574865, rs8179673, and rs10181656) were associated with susceptibility to T1D. When we stratified T1D patients into early onset and late-onset subgroups on the basis of fewer or more than 7.8 years of age at diagnosis, however, the minor alleles of three SNPs (rs7574865, rs8179673, and rs10181656) showed a significant association with susceptibility to T1D in the early-onset subgroup (i.e., rs7574865, OR = 1.44 [1.03-2.01], P < 0.05), but not in the late-onset subgroup, suggesting that STAT4 is related to earlier development of T1D. The analysis of genotypes and haplotypes of the same SNPs (rs7574865, rs8179673, and rs10181656) showed very comparable degrees of risk for T1D. The age at diagnosis is lowest in the patients carrying the homozygotes of a minor allele, middle in the heterozygotes, and highest in the homozygotes of a major allele, suggesting the dosage effects of risk alleles on the age of onset of disease. Recognizing that only the early-onset cases might represent the true autoimmune T1D in Asian populations, we see that STAT4 alleles and haplotype might influence cytokine signaling and, therefore, development of T1D. PMID- 19120277 TI - Molecular scanning of the gene for thioredoxin, an antioxidative and antiapoptotic protein, and genetic susceptibility to type 1 diabetes. AB - Oxidative stress has been implicated in the destruction of beta cells in type 1 diabetes (T1D). Thioredoxin has been shown to protect cells from oxidative stress and apoptosis. In this study, we screened for sequence variants of the human thioredoxin gene (TXN), and studied the association of the variants in persons with T1D in Japanese. The frequency of the A allele of the G/A SNP in the 3' flanking region was highest in T1D (8.4%), followed by type 2 diabetes (6.8%), and the lowest in the controls (5.9%), suggesting the contribution of TXN polymorphism to susceptibility to T1D. PMID- 19120276 TI - From Type 1, through LADA, to type 2 diabetes: a continuous spectrum? AB - This study aims to explore whether diabetes mellitus from T1DM, through LADA, to T2DM presents a continuous spectrum in terms of HLA-DQ genetic background. We recruited 223 patients with autoimmune T1DM, 215 with LADA, 206 with T2DM, and 228 nondiabetic controls, and then defined their HLA-DQA1 and -DQB1 genotypes and haplotypes. T1DM patients were divided into two groups depending on age of onset of disease: juvenile-onset (JO; before the age of 20) and adult-onset (AO; after the age of 20). LADA patients were sorted according to the GADA titer: the LADA1 group had titers higher than 175 U/mL, whereas the LADA2 group had lower titers. The susceptible haplotypes of T1DM were DQA1*03-DQB1*0303, DQA1*03-DQB1*0401, and DQA1*05-DQB1*0201. The protective haplotype was DQA1*0102-DQB1*0602. The frequency of DQA1*03-DQB1*0303 in JO, AO, LADA1, LADA2, T2DM, and control groups were 38.2%, 34.2%, 25.3%, 18.9%, 17.5%, and 16.5%, respectively. The frequencies of DQA1*05-DQB1*0201 were 21.2%, 15.0%, 12.7%, 4.6%, 3.6% and 3.3%; the frequencies of DQA1*03-DQB1*0401 were 11.3%, 9.4%, 11.3%, 5.4%, 4.4% and 3.3%; and the frequencies of DQA1*0102-DQB1*0602 were 1.4%, 1.7%, 0.7%, 4.6%, 7.0%, and 5.3%. The linear-by-linear association showed that the frequency of DQA1*03 DQB1*0303, DQA1*05-DQB1*0201, and DQA1*03-DQB1*0401 presented a decremental tendency in JO, AO, LADA1, LADA2, T2DM, and control groups. The preliminary data demonstrated that the susceptible haplotypes of the HLA-DQ gene present a continuous spectrum from typical T1DM, through LADA, to T2DM, which deserves further investigation. PMID- 19120278 TI - Risk conferred by HLA-DR and DQ for type 1 diabetes in 0-35-year age group in Sweden. AB - HLA DR4-DQ8 and DR3-DQ2 haplotypes account for 89% of newly diagnosed cases of type 1 diabetes (T1D) in Sweden. The presence of a single copy of DQ6 confers protection. The aim of the present study is to evaluate whether the risk conferred by high risk HLA DR and DQ to T1D is similar in all regions of Sweden and see whether there are any significant regional differences. The subjects comprised 799 consecutively diagnosed T1D patients and 585 age-, sex-, and geography-matched healthy controls in the age group 0-35 years. HLA typing for high-risk haplotypes was previously performed using PCR-SSOP and RFLP. The results showed that HLA DR3-DR4 gave an odds ratio of 8.14 for the whole of Sweden. However, when the study group was divided into six geographical regions, subjects from Stockholm had the highest OR, followed by those from Lund, Linkoping, Gothenburg, Umea, and Uppsala. Absolute protection was conferred by the presence of DQ6 in subjects from the Linkoping region, but varied in the other regions. The frequency of DR3 and DQ2, DR4 and DQ8, DR15, and DQ6 in patients showed high linkage for each region, but were different between regions. IN CONCLUSION: The risk conferred by high-risk HLA varies in different regions for a homogenous population in Sweden. The results highlight the important role played by the various environmental factors in the precipitation of T1D. PMID- 19120279 TI - Closing the circle between the bedside and the bench: Toll-like receptors in models of virally induced diabetes. AB - Animal models provide many strategies to unravel the complex interplay of genetic, immunologic, and environmental factors involved in the pathogenesis of type 1A (autoimmune) diabetes. Diabetes can be studied at multiple levels, and new technological advancements provide insights into the functioning of organelle and cellular structures. The role of innate immunity in the response to environmental pathogens has provided possible biochemical and molecular mechanisms which can explain certain clinical events in diabetes. These investigations may uncover new therapies and strategies to prevent type 1A diabetes. PMID- 19120280 TI - Toll-like receptors and diabetes. AB - Toll-like receptors (TLRs) recognize molecular patterns relating to a variety of microbial infections. Stimulation through TLRs leads to activation of antigen presenting cells, production of inflammatory cytokines creating inflammation, and production of type 1 interferons (IFNs) that include IFN-alpha and -beta, and exerts direct effects on regulatory cells. These effects can direct the immune response, dealing with the immediate problems of infection and activating more specific responses of the adaptive immune system. However, it has recently been recognized that these receptors may recognize endogenous ligands that include DNA, RNA, and proteins that arise from cellular stress. This may have an effect on autoimmune responses in a number of ways, both activating and inhibitory. The means by which infection or endogenous stimuli through TLRs may influence autoimmunity will be discussed. PMID- 19120281 TI - Different KIRs confer susceptibility and protection to adults with latent autoimmune diabetes in Latvian and Asian Indian populations. AB - KIRs (killer Ig-like receptors) expressed on natural killer (NK) cells are an important component of innate (and adaptive) immunity. They are either activatory or inhibitory, and certain KIRs are known to interact with specific motifs of HLA Class I molecules, which is very crucial in determining whether a cell is targeted to lysis or otherwise. Latent autoimmune diabetes in adults (LADA) is a slowly progressive form of autoimmune diabetes, with an adult onset (>30 years). Because autoantibodies and autoimmunity involved are involved in the etiology of LADA, KIRs might play an important role in conferring susceptibility to or protection against the disease. The purpose of this study was to identify killer immunoglobulin-like receptor (KIR) genes, which are associated with susceptibility to and protection against type 1 diabetes in Latvian and Asian Indian patients with LADA. KIR and HLA-C ligand genotyping was performed using PCR-SSP in LADA patients from Latvia (n= 45) with age- and sex-matched controls (n= 92) and from India (n= 86) with controls (n= 98). Results showed that in Latvian patients with LADA, KIRs 2DL1, 2DS2, and 2DS4 were associated with susceptibility and KIR 2DS5 with protection. In Asian Indian LADA patients, KIRs 2DL5 and 3DL1 were associated with susceptibility and KIRs 2DS1 and 2DS3 with protection. Stratification analyses for KIRs that bind to HLA-C1 and C2 were performed. We concluded that KIRs are important in conferring susceptibility (or protection) to adult patients with LADA in both our study populations. However the KIR genes (and their HLA-C ligands) conferring susceptibility or protection in these two populations differ, showing a role of ethnicity in disease susceptibility. PMID- 19120283 TI - The distribution of adiponectin receptors on human peripheral blood mononuclear cells. AB - Adiponectin, an adipocytokine with anti-inflammatory and insulin-sensitizing properties, may provide a mechanism by which insulin resistance accelerates autoimmunity in type 1 diabetes (T1D). Its actions are mediated by two receptors, adiponectin receptors 1 (ADIPOR1) and 2 (ADIPOR2). In this study, we measured their distribution on human peripheral mononuclear cells by flow cytometry. ADIPOR1 is present approximately on 1% of T cells, 93% of monocytes, 47% of B cells, and 21% of NK cells (P < 0.01 for difference between subsets). The distribution of ADIPOR2 was found to be similar (r= 0.992, P < 0.01), and staining could be blocked in an antigen-specific manner. We were also able to confirm our finding at an RNA level by PCR using sequence-specific primers. Our data are consistent with an immunoregulatory role for adiponectin in T1D. PMID- 19120282 TI - Innate immune pathways in virus-induced autoimmune diabetes. AB - We recently hypothesized that Toll-like receptor-induced innate upregulation by Kilham rat virus (KRV) in the BioBreeding diabetes-resistant (BBDR) rat model plays a key role in the mechanism of diabetes induction. To address this hypothesis, we analyzed innate immune signaling pathways upregulated by KRV in vitro and in vivo. We demonstrate that KRV activates the signal transducer and activator of transcription (STAT)-1 in spleen cells in vitro and that this activation can be blocked by TLR9 inhibition. We also show that KRV upregulates STAT-1 in pancreatic lymph nodes early after virus infection. Our data may implicate TLR9-induced STAT-1 signaling pathways in KRV-induced innate immune activation in the BBDR rat and raise the possibility that these pathways are involved in mediating autoimmune diabetes. PMID- 19120284 TI - The role of Toll-like receptors 3 and 9 in the development of autoimmune diabetes in NOD mice. AB - Innate immunity is mediated, at least in part, through a number of receptors known as Toll-like receptors (TLRs), which are activated by different microbial stimuli. Adaptive immunity, including autoimmunity, follows the innate response in a more specific manner. To investigate the roles of TLR3 and TLR9 in the development of type 1 diabetes, we generated NOD mice that are deficient in TLR3 and 9, respectively. There was no obvious difference in the incidence of spontaneous diabetes between TLR3-deficient mice and TLR3 heterozygous mice. However, TLR9-deficient mice were markedly protected from the disease compared to TLR9 heterozygous mice. Our results suggest that different TLRs play a varying role in autoimmune diseases. PMID- 19120285 TI - TLR agonists abrogate co-stimulation blockade-induced mixed chimerism and transplantation tolerance. AB - We investigated the mechanisms by which Toll-like receptor (TLR) agonists affect the induction of mixed chimerism and skin allograft survival in mice treated with co-stimulation blockade (CB). We report that TLR agonists prevent the generation of mixed chimerism by breaking tolerance in the alloreactive CD4(+) and CD8(+) T cell compartments, and that type I interferon (IFN) is important in this process. Understanding how environmental perturbations affect CB-induced transplantation tolerance may lead to more effective regimens that can be used as an approach for the treatment of type I diabetes, for which the transplantation of pancreatic islets is a promising therapy. PMID- 19120286 TI - Exendin-4 therapy in NOD mice with new-onset diabetes increases regulatory T cell frequency. AB - Recent studies, albeit controversial, have suggested that the incretin exendin-4 (Ex-4) is capable of inducing beta cell proliferation in vivo. Furthermore, this compound has been shown to enhance the ability of other agents (e.g., anti-CD3, antilymphocyte serum) to reverse type 1 diabetes (T1D) in NOD mice. However, the mechanisms underlying this beneficial action for disease reversal remain largely unclear. Herein, we tested the hypothesis that Ex-4 therapy may act as a stimulator of regulatory T cells (Tregs). We evaluated the effect of Ex-4 (Byetta; 0.2 microg/mouse/day for 30 days) treatment on the frequency and function of Tregs and changes in the cytokine profile of NOD mice with recently diagnosed T1D. In comparison to that of saline-treated control NOD mice, the frequency of Tregs was increased in Ex-4-treated mice. Suppression assays demonstrated a trend towards increased Treg suppression after administration of Ex-4, but were limited by small sample size. Lastly, Ex-4 treatment induced production of IL-10, indicating a possible shift towards a more Th2-like phenotype. Taken collectively, these data suggest that in addition to its potential effects on beta cell proliferation, Ex-4 may also act as a regulator of the immune response. PMID- 19120287 TI - Role of increased ROS dissipation in prevention of T1D. AB - Protection of pancreatic beta cells is an approach to prevent autoimmune type 1 diabetes (T1D) and to protect transplanted islets. Reactive oxygen species (ROS) are important mediators of beta cell death during the development of T1D. We have examined the role of elevated ROS dissipation in the prevention of T1D using the ALR mouse strain. The selection of ALR, for resistance against alloxan-induced free radical-mediated diabetes, led to a strain of mice with an elevated systemic as well as pancreatic ROS dissipation. Independent genetic mapping studies have identified ALR-derived diabetes protective loci. Conplastic and congenic mouse as well as cell line studies have confirmed the genetic mapping and demonstrated that the elevated ROS dissipation protects ALR beta cells from autoimmune destruction. Our data support the hypothesis that elevated ROS dissipation protects beta cells against autoimmune destruction and prevents T1D development. PMID- 19120288 TI - Apoptotic non-beta cells suppress beta cell antigen-reactive T cells and induce beta cell antigen-specific regulatory T cells. AB - Steady-state cell apoptosis plays an important role in maintenance of self tolerance. Based on this notion, the use of apoptotic cells to restore self tolerance to beta cell antigens is a rational approach to type 1 diabetes (T1D) prevention. Our previous study demonstrated that transfusion of apoptotic beta cells induced immune tolerance to beta cell antigens in NOD mice. However, concerned about the limited beta cell source for future clinical applications, we attempted in the present study to develop a more practical approach for T1D prevention using apoptotic non-beta cells. We found that UVB-irradiation-induced apoptotic NOD splenic stromal cells significantly suppressed beta cell antigen specific T cell proliferation in vitro and in vivo. Furthermore, TCR-transgenic CD4(+) T cells primed by the antigens to which they were specific in the presence of UVB-irradiated stromal cells were rendered unresponsive to the antigen restimulation, a result that was partially attributed to the induced IL-10 producing regulatory T cells. Of more interest, transfusion of UVB-irradiated stromal cells appeared to induce beta cell antigen-responding IL-10-producing regulatory T cells in vivo. Most importantly, transfusion of UVB-irradiated stromal cells effectively prevented T1D in NOD mice, which is consistent with these findings. This study suggests that it is possible to use apoptotic non-beta cells such as peripheral blood mononuclear cells to induce beta cell antigen specific tolerance, thereby preventing T1D in humans. PMID- 19120289 TI - Persistence of residual beta cells and islet autoimmunity during increasing duration of diabetes in NOD mice and experimental approaches toward reversing new onset disease with bioactive peptides. AB - The precise fate of beta cells and the presence of islet infiltrates after onset of type 1 diabetes have not yet been fully characterized. Recently we showed that in newly diabetic NOD mice an appreciable number of beta cells remain. This was also observed during the first 2 weeks of diabetes in NOD mice without treatment with insulin. However, the mean number of beta cells per unit islet cross sectional area decreased with increasing duration of disease. In contrast, glucagon and somatostatin cell numbers showed an increase. The persistence of insulitis in several islets until 4 weeks of diabetes suggests ongoing beta cell autoimmunity over a protracted phase. Combined daily treatment of newly diabetic NOD mice with epidermal growth factor (EGF) and gastrin for the first 14 days of diabetes resulted in temporary restoration of normoglycemia in 7 of 15 mice. We speculate that the residual beta cells present soon after onset of diabetes may respond to experimental regeneration. Treatment of newly diabetic NOD mice with the bioactive peptides EGF and gastrin resulted in partial and temporary reversal of diabetes. We propose that peptide therapies combined with other benign immunomodulatory approaches to rescue and preserve beta cells in the long term and to prevent recurring autoimmunity may be more effective than peptide therapy alone in reversing diabetes in NOD mice. PMID- 19120290 TI - TGFbeta plasmid construction and delivery for the prevention of type 1 diabetes. AB - Studies of animals with spontaneous autoimmune diabetes have revealed that autoreactive T cells that mediate islet beta cell destruction can be manipulated by the administration of Th(2) cytokines. Using gene delivery to express the targeted protein, we can overcome the need for frequent administration of cytokines on account of their short half-lives. In this study, the effect of hTGFbeta gene delivery was evaluated both in vitro and in vivo using an adenovirus vector (Ad) constructed with an hTGFbeta cDNA. In vitro transfection assays of the construct in HepG2, beta cell lines, and islets showed good expression levels of hTGFbeta and activation of smad3. Ad-hTGFbeta enhanced differentiation and proliferation in the beta cell line or islets without causing apoptosis. Of interest, Ad-hTGFbeta transduction in CD4(+)CD25(-) T cells resulted in a significant enhanced expression of CD25 and a regulatory T cell specific transcription factor, Foxp3. To evaluate in vivo efficacy, Ad-hTGFbeta was intravenously injected into 7-week-old NOD mice and compared to the transduction using the vector only. The Ad-hTGFbeta group had persistent gene expression for longer than 5 weeks, and high TGFbeta serum level was secreted. There was no difference in the degree of insulitis between the Ad-hTGFbeta group and controls. Although we found favorable in vitro results, such as decrease in islet apoptosis, enhanced proliferation and differentiation, and increase in the level of CD4(+)CD25(+) regulatory T cells, there was no difference in reduction of the development of T1D between controls and Ad-hTGFbeta-injected mice. Nevertheless, if we find the appropriate mode and timing of TGFbeta gene transduction, Ad-hTGFbeta gene therapy might be useful in therapeutic cytokine delivery for the treatment of T1D. PMID- 19120291 TI - Intravenous administration of proinsulin 1 or 2-expressing fiber-mutant recombinant adenovirus vector protects against the development of diabetes in NOD mice. AB - Insulin has been reported as a major autoantigen in both human and murine type 1 diabetes (T1D). Insulin1-knockout NOD mice with only insulin2 are protected against the development of autoimmune diabetes, suggesting that insulin1 has strong immunogenicity and insulin2 has weak immunogenicity or a possible protective role in the pathogenesis of type 1 diabetes. In this study, we have developed fiber-mutant adenovirus vectors that express murine proinsulin1 or proinsulin2 (named Ad.Pins1-RGD/Ad.Pins2-RGD) and administered those virus vectors to the NOD mouse to evaluate modulation of autoimmune responses. The intravenous administration of either Ad.Pins1-RGD or Ad.Pins2-RGD at 3 and 5 weeks of age strongly suppressed the development of overt diabetes, accompanied by a significant reduction of insulin autoantibody (IAA), and suppression of disease was similar between administration of Ad.Pins1-RGD and that of Ad.Pins2 RGD. Our study suggests that systemic administration of fiber-mutant adenovirus vectors, which induce transient expression of proinsulin, may be applicable to a gene therapy inducing tolerance to insulin. PMID- 19120292 TI - IFN-alpha can both protect against and promote the development of type 1 diabetes. AB - Viral infection has been hypothesized to be one of the environmental triggers for the development of type 1 diabetes. Infection induces a large amount of interferon-alpha (IFN-alpha) produced by dendritic cells and other cells. To test the role of IFN-alpha in the development of diabetes, we have used three different experimental approaches in both diabetes-prone and -resistant animal models for type 1 diabetes. Our results suggested that a viral mimic or IFN-alpha can either suppress or promote the development of autoimmune diabetes, depending on the model system. It is likely that IFN-alpha plays a complex role in the etiology of type 1 diabetes. PMID- 19120293 TI - Update on worldwide efforts to prevent type 1 diabetes. AB - This paper reviews worldwide efforts to interdict the type 1 diabetes (T1D) disease process, during the stage of evolution of the disease prior to the time of disease onset. The goal of intervention before disease onset is to arrest immune destruction and thus prevent or delay clinical disease. In this regard, there have been several large-scale multicenter randomized controlled clinical trials designed to prevent T1D. These have tested nicotinamide, parenteral insulin, oral insulin, nasal insulin, and the elimination of cow's milk from infant feeding. PMID- 19120294 TI - Tobacco exposure and diabetes-related autoantibodies in children: results from the ABIS study. AB - Passive smoking has decreased in recent years ("increased hygiene"). Less environmental tobacco smoke (ETS) gives increased hygiene that, if the hygiene hypothesis is true, in turn might give more autoimmune diseases. The presence of auto antibodies is considered to be an early indicator of type 1 diabetes (T1D). Because tobacco exposure may influence the immune system, we analyzed the relation between passive smoking and development of autoantibodies. A subsample (n= 8794) of the children in the ABIS study was used for this analysis. The parents answered questionnaires on smoking from pregnancy and onwards, and blood samples from the children aged 2.5-3 years were analyzed for GADA and IA-2A. Results showed that there was no significant difference in the prevalence of GADA or IA-2A (>95 percentile) between tobacco-exposed and nonexposed children. It was concluded that passive smoking does not seem to influence development of diabetes related autoantibodies early in life. PMID- 19120295 TI - Variations in incidence of type 1 diabetes in different municipalities of stockholm. AB - This article reports a test of the hypothesis that municipalities within the County of Stockholm have varying incidence rates of type 1 diabetes (T1D), suggesting a strong etiologic environmental component to the disease. The study group included T1D patients in the age group from birth to 18 years who were diagnosed each year from 20 municipalities in Stockholm County during the 1990 2003. Specific incidence rates by age, sex, and socioeconomic characteristics (income level, proportion of taxpayers, proportion of foreigners, population density and green cover) were estimated annually together with age standardization. chi(2) analyses were used for the statistical assessment of variability in incidence. During the study period, 733 newly diagnosed T1D patients aged 0-18 years were recorded from the 20 municipalities under study. The overall age-standardized incidence in these 20 municipalities was 24.38 (22.65-26.21) per 100,000, with 45.35 (32.08-62.29) as highest and 13.41 (9.53 18.35) as lowest estimated incidence. For all socioeconomic variables statistically significant heterogeneity was demonstrated in the standardized incidence rate. High green index was positively associated with the incidence of T1D, as was low population density. For the three remaining socioeconomic variables no clear patterns of associations with incidence of T1D were seen. This study demonstrates a considerable and statistically significant variation between the lowest and highest values in the incidence and prevalence rates for T1D in municipalities of Stockholm County. Such variation seems unlikely to be explained by genetic differences since the population is homogeneous. Our study provides support for the hypothesis that environmental factors have a major influence on the pathogenesis of T1D. PMID- 19120296 TI - Eradication of Helicobactor pylori may trigger onset of type 1 diabetes: a case report. AB - In patients with idiopathic thrombocytopenic purpura (ITP), which is considered an autoimmune disease, eradication of Helicobacter pylori cures a significant number of patients. However, here we report an adult patient who developed type 1 diabetes (T1D), which is also considered an autoimmune disease, after eradication of Helicobacter pylori. Although further investigation is needed to understand the pathophysiology of T1D, we would like to emphasize that clinicians should consider the risk of its development after eradication of Helicobactor pylori. PMID- 19120297 TI - Diabetogenic effects of the most prevalent enteroviruses in Finnish sewage. AB - Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes (T1D) and also sometimes precipitate the clinical disease. We have recently demonstrated that (1) enterovirus-positive islet cells were seen on postmortem pancreatic specimens of several T1D patients but not in the corresponding samples of nondiabetic controls, and (2) several different enteroviruses can be associated with T1D. Enterovirus infections are transmitted from person to person by fecal-oral or respiratory routes, which means that infections usually start from the respiratory or gastrointestinal mucosa. Regardless of the clinical symptoms of the disease, viral replication continues in the submucosal lymphatic tissue for several weeks, up to a couple of months, and during that time the virus is excreted into the feces and translocated to the environment. Monitoring of sewage samples for enteroviruses can be used as a tool in epidemiologic studies of enterovirus. Finland has successfully used environmental control data in poliovirus surveillance for decades. About 24 samples have been collected annually from the Helsinki region, which covers about 20% of the population. In the present study, we have reanalyzed the sewage samples of the years 1993-2004 for nonpolio enteroviruses by inoculating them into five different continuous cell lines known to cover a wide range of serotypes. Isolated strains were identified by RT-PCR and VP1 sequencing. The most commonly detected serotypes were coxsackie B viruses (CBV1 5) and echoviruses (E6, 7, 11, 25, 30). Diabetogenic effects of the most prevalent enterovirus serotypes were studied in primary human beta cells. PMID- 19120298 TI - Co-transplantation of stromal cells interferes with the rejection of allogeneic islet grafts. AB - Side effects associated with current immunosuppressive therapy complicate the use of islet transplantation as a treatment for type 1 diabetes. Multipotent mesenchymal stromal cells (MSCs) have demonstrated immunomodulatory activity. The purpose of this study was to investigate whether a murine stromal cell line affects graft rejection in a fully major histocompatibility complex (MHC) mismatched islet transplant model. We show that stromal cells have an inhibitory effect on T cell proliferation in vitro, and that they slow down the rejection of allogeneic islets. These findings indicate a possibility to use MSCs as a treatment to prolong the survival of islet grafts. PMID- 19120299 TI - Anti-CD20 treatment prolongs syngeneic islet graft survival and delays the onset of recurrent autoimmune diabetes. AB - Type 1 diabetes is an autoimmune disease characterized by T cell-mediated destruction of pancreatic islet beta cells. Pancreatic islet transplantation with long-term immunosuppressive drug treatment is an accepted therapeutic option for patients with type 1 diabetes suffering from disabling hypoglycemia on insulin treatment. Here we investigated the replacement of immunosuppressive drug treatment with immune tolerance establishment induced by temporary B cell depletion therapy for islet transplantation. The result suggested that the combined therapy of B cell depletion and syngeneic islet transplantation may reverse the disease in hCD20/NOD mice. PMID- 19120300 TI - Autologous hematopoietic stem cell transplantation for type 1 diabetes. AB - In this review, we present (1) the scientific basis for the use of high-dose immunosuppression followed by autologous peripheral blood hematopoietic stem cell transplantation for newly diagnosed type 1 diabetes (T1D); (2) an update of the clinical and laboratory outcome of 20 patients transplanted at the University Hospital of the Ribeirao Preto Medical School, University of Sao Paulo, Brazil, and followed up to January/2008, including 4 relapses among 19 patients without previous ketoacidosis; (3) a commentary on criticisms to our article that appeared in four articles from the scientific literature; and (4) a discussion of the prospectives for cellular therapy for T1D. PMID- 19120301 TI - Time course of islet loss after intraportal transplantation. AB - A high percentage of islets fail to survive intraportal transplantation, but the absolute amount and time course of this loss are uncertain. We have devised a technique to directly quantitate the number of surviving islets using simultaneous selective transplantation of islets and inert beads into the posterior lobes of the liver. Islet:bead ratio did not change significantly within the first 2 hours, but fell progressively thereafter, giving calculated islet survival rates of 89, 43, and 66% at days 1, 7 and 28, respectively. This technique provides a baseline from which to develop strategies to improve the survival of intraportally transplanted islets. PMID- 19120302 TI - Human regulatory CD8 T cells. AB - Administration of a humanized monoclonal anti-CD3 antibody (mAb) to patients with type 1 diabetes (T1D) increases their C-peptide responses and the CD8/CD4 ratio. Incubation of human peripheral blood mononuclear cells (PBMC) with mAb in vitro has been shown to induce CD8(+) regulatory T cells (Tregs) capable of inhibiting proliferation of CD4(+) T cells. We hypothesized that CD8(+) Tregs function through secretion of cytokines. To test that possibility, we generated CD8(+) Tregs, sorted them by FACS, incubated them with syngeneic CD8-depleted PBMC in the presence of staphylococcal enterotoxin B (SEB), and measured proliferation of T cells and cytokines. Using neutralizing anti-cytokine mAbs, we show that the inhibitory effect of CD8(+) Tregs could be partially alleviated by anti-CCL-4, anti-TNF, and to a lesser extent anti-IL2, suggesting that these cytokines contribute to CD8(+) Treg function. PMID- 19120304 TI - Conserved epitopes in the protein tyrosine phosphatase family of diabetes autoantigens. AB - IA2 and phogrin are important targets of humoral and cell-mediated autoimmunity in type 1 diabetes in man. They belong to a conserved subfamily of transmembrane protein tyrosine phosphatases (PTPs) associated with the regulatory pathway of secretion. To examine potential cross-reactivity between PTP family members we tested sera from T1D patients for reactivity to IA2, and the Drosophila (FLYDA) and C. elegans (IDA) orthologs using radioimmunoprecipitation assays of (35)S Met labeled in vitro translated products of the cytosolic domains of these proteins. Approximately 80% of sera reacted with at least one probe. Of these, 82.5% showed reactivity to human IA2, 74.1% to FLYDA, and 33.7% to IDA. The majority of sera that bound FLYDA and/or IDA also recognized IA2. This raises the possibility that in some cases reactivity to IA2 may have arisen by molecular mimicry. PMID- 19120303 TI - Type 1 diabetes versus type 2 diabetes with onset in persons younger than 20 years of age. AB - Type 1 diabetes (T1D) is the most common form of diabetes in children in Western countries. There have been no large studies of childhood diabetes from India. We undertook the MEDI study (Multicenter Survey of Early Onset Diabetes in India) to assess the proportion of various subtypes of diabetes among the young subjects presenting to the endocrinology divisions of seven large teaching hospitals in different regions of India. In addition, we compared the clinical features of T1D and type 2 diabetes (T2D) in Indian subjects. Patients with onset of disease at younger than 20 years of age were included in this study. Six hundred and three subjects (603) were studied of whom 535 subjects (89%) had T1D, 36 (6%) had T2D, 18 (3%) had diabetes related to tropical pancreatitis or other forms of chronic pancreatitis, while other subtypes accounted for the rest. Compared to those with T2D, subjects with T1D were younger, had a lower C-peptide level, higher prevalence of ketosis, lower prevalence of acanthosis nigricans, and lower LDL and triglyceride levels. When compared with that of T2D, a higher proportion of patients with T1D were positive for GAD-65 and IA-2 antibodies, and this difference was statistically significant for GAD-65 antibodies. Overall, this large multicenter study showed that T1D is the commonest form of diabetes in childhood. T2D is the next most common kind, while chronic pancreatitis-related diabetes is uncommon. PMID- 19120305 TI - Humoral immune response to islet autoantigens in Japanese patients with type 1 diabetes. AB - In this study, we evaluated autoantibodies to IA-2 (IA-2As), glutamic acid decarboxylase 65 (GADAs), and islet cell antibodies (ICAs) in 233 patients with type 1 diabetes (M:F = 90:143, mean duration 4.0 +/- 6.7 yr) as a cross-sectional study. Of 233 patients with type 1 diabetes, IA-2A was detected in 58% of patients with duration within 2 weeks, 61% of patients with duration <1 yr, 41% of patients with diabetes for 1-3 yr, 29% for 4-9 yr, and 21% for >or=10 yr. These prevalences were similar to those of ICA, while the prevalence of GADA was not influenced by duration of diabetes with positivity of 63-74%. Thus, as the duration of diabetes became longer, the frequency of GADA(+)/IA-2A(-) patients increased and the frequency of GADA(+)/IA-2A(+) patients decreased. However, the frequency of GADA(-)/IA-2A(+) patients was not influenced by duration of diabetes. The prevalence of IA-2A was significantly higher in abrupt-onset group (68%, n= 79) compared to the slowly progressive group (23%, n= 22) in new-onset patients (P= 0.0001). However, there was no difference in the IA-2A frequency between these two groups (abrupt-onset 26%, n= 53 vs. slowly progressive 24%, n= 21) in patients with long-standing disease, suggesting that IA-2A positivity might persist in patients with slowly progressive type 1 diabetes. These results emphasize the heterogeneity of humoral autoimmunity to protein tyrosine phosphatase-like molecules, but not to GAD, in patients with type 1 diabetes. PMID- 19120306 TI - Identification of a major humoral epitope in Slc30A8 (ZnT8). AB - The human zinc transporter Slc30A8 (ZnT8) is a major target of humoral autoimmunity in human type 1A diabetes. However, despite extensive conservation, the majority of human autoimmune sera fail to recognize the murine ortholog. Moreover, Slc30A8 appears not to be a significant target of humoral autoimmunity in the NOD mouse. We therefore "humanized" the murine protein by site-directed mutagenesis. Only conversion of Q324 to arginine (equivalent to R325 in the human protein) partially restored reactivity to a pool of sera selected for high titers to the human probe. Additionally, the reciprocal mutation (human R325 to Q) abolished reactivity for 38/103 (36.9%) of ZnT8(+) sera. We conclude that the C terminal domain of human ZnT8 contains at least two discrete epitopes, one of which is critically dependent upon the arginine residue at position 325. PMID- 19120307 TI - SlC30A8 is a major target of humoral autoimmunity in type 1 diabetes and a predictive marker in prediabetes. AB - Type 1A diabetes (T1D) results from autoimmunity targeted at a limited number of molecules that are expressed in the pancreatic beta cell. Putative novel autoantigen candidates were identified from microarray expression profiling of human and rodent islet cells. The highest ranking candidate was Slc30A8 (zinc transporter 8; ZnT8), which was screened by radioimmunoprecipitation assays against new-onset T1D and prediabetic sera. Such assays detected 63% of subjects with new-onset diabetes, but fewer than 2% of controls, 3% of those with type 2 diabetes, and 10% of patients with other autoimmune disorders. ZnT8 autoantibodies were found, however, in 26% of T1D subjects previously classified as autoantibody-negative on the basis of existing markers (GADA, IA2 A, IAA, and ICA). We conclude that SLC30A8 provides an important additional and independent predictive marker for T1D. PMID- 19120308 TI - Prevalence of organ-specific autoantibodies in childhood- and adult-onset type 1 diabetes. AB - There are no studies that compare the prevalence of organ-specific autoantibodies (OSAs) between adult (>or= 16 years) and childhood-onset type 1 diabetes (T1D). We evaluated the prevalence of the following OSAs: thyroid peroxidase, thyroid receptor, parietal cell, intrinsic factor, tissue transglutaminase, adrenal cortex, mitochondrial, smooth muscle, liver kidney microsomal, and ovarian autoantibodies. Three hundred twenty-seven (327) adults were screened for one or more of these OSAs. The prevalence of all the OSAs studied was similar in both groups. The most prevalent OSA observed was tissue transglutaminase (childhood onset disease = 14.3%; adult-onset disease = 13.6%). This study did not demonstrate a distinct difference in the prevalence of OSAs between adult- and childhood-onset T1D patients. PMID- 19120309 TI - Carboxypeptidase-H autoantibodies differentiate a more latent subset of autoimmune diabetes from phenotypic type 2 diabetes among Chinese adults. AB - This study aimed to investigate whether carboxypeptidase-H antibody (CPH-Ab) can help identify latent autoimmune diabetes in adults (LADA). Phenotypic type 2 diabetic (T2D) patients (n= 1296) were studied for CPH Abs and autoantibodies to glutamic acid decarboxylase (GAD-Abs). CPH-Ab(+) T2D patients also underwent testing for insulinoma protein tyrosine phosphatase (IA-2A). Clinical features were compared among CPH-Ab(+), GAD-Ab(+), and Ab(-) T2D patients. Some of the antibody-positive patients were followed up for 3 years to assess beta cell function. The prevalence of CPH-Abs in T2D patients was 4.8%, significantly higher than that in controls. Double positivity was rare between CPH-Abs and GAD Abs or IA-2A. Compared to patients with Ab(-) T2D, those with CPH-Ab(+) T2D had lower BMI, lower fasting C-peptide (FCP) levels, and more frequent ketosis, while not as much as did those with GAD-Ab(+) T2D. The mild beta cell dysfunction in patients with CPH-Ab(+) T2D was associated with their longer duration of diabetes. No marked change of C-peptide in the CPH-Ab(+) group was found during follow-up. These findings demonstrated that CPH-Abs may allow discrimination of a more latent subset of adult-onset autoimmune diabetes (LADA) whose features are intermediate between those with classic GAD-Ab(+) LADA and patients with Ab(-) T2D. PMID- 19120310 TI - Latent autoimmune diabetes in an adult. AB - We report a case of latent autoimmune diabetes in adults (LADA), also known as slowly progressive type 1 diabetes (SPT1D), followed up for changes, including reactivity to the GAD65 antibody epitope for the 9-year period from impaired glucose tolerance (IGT) to the insulin-dependent stage (IDDM). This 69-year-old male was identified as having IGT by health checkup in 1998. As he was GAD65 positive (high titer), we initiated close clinical follow-up. In 2003, a 75-g oral glucose tolerance test showing a diabetic pattern confirmed that he had progressed to diabetes. During this period, fasting plasma glucose remained within normal range and insulin secretion was unchanged compared to that at the time of IGT diagnosis. His fasting plasma glucose and HbA1c levels began to increase in 2004 and serum C-peptide began to decrease in 2005. Insulin treatment was started in August 2006. GAD65 antibody titer was high (13900 U/ml) in 1998 and has remained high throughout follow-up. The patient's GAD65 antibodies were initially directed to the b96.11-defined epitope only, recognized as an indicator of T1D-like pathogenesis in our former study. During follow-up, he developed reactivity to more epitopes (MICA-3 and MICA-4, DPC, and DPA). The course of this case suggests that the b96.11-defined epitope is important for distinguishing LADA patients who progress to IDDM from those who do not and that epitope maturation is restricted to LADA patients who progress to IDDM, an observation similar to that in children at risk for developing typical IDDM. PMID- 19120311 TI - The sentinel role of CD8 T cells in regulating CD4 T cell responses to proinsulin in beta-islet cell autoimmunity. AB - Circulating CD4 T cells specific for peptide epitopes of proinsulin and other autoantigens are markers of autoimmune beta cell destruction in type 1 diabetes, while the role of CD8 T cells is still largely unknown. Here we show that CD8 T cells of a diabetic patient--after rechallange with proinsulin peptides--secrete IFNgamma and granzyme B, markers of their effector capacity. On the other hand, CD8 T cells of the same patient in a "cross-talk" with proinsulin-specific CD4 T cells suppress their proliferation. If confirmed in larger numbers of subjects with beta-islet cell autoimmunity, these results may help us to understand the role of CD8 cells in disease progression and extend our knowledge of disease pathogenesis. PMID- 19120312 TI - Diverse foxp3 expression in children with type 1 diabetes and celiac disease. AB - Imbalance between different types of T lymphocytes, such as T helper (Th) and regulatory T cells (Tregs), has been reported to play a part in the pathogenesis behind such autoimmune diseases as type 1 diabetes (T1D) and celiac disease (CD). Defects in Tregs are proposed to at least partly explain the imbalance of Th cells found in children with immunologic diseases. Peripheral blood mononuclear cells from 24 children with T1D and/or CD, and reference children (that is, those without any of these diseases) were stimulated with disease-associated antigens (insulin, gluten, transglutaminase [tTG]), and phytohemagglutinin (PHA). The mRNA expression of the Treg-associated marker FOXP3 was analyzed with multiplex real time RT-PCR. Children with T1D showed both a low spontaneous (P < 0.05) and PHA induced (P < 0.01) expression of FOXP3 mRNA compared to children with CD. Children with T1D also had a low PHA-induced FOXP3 mRNA expression compared to the group of children diagnosed with both T1D and CD (P < 0.05). Spontaneous (P < 0.05) and PHA-induced (P < 0.05) FOXP3 mRNA expression was high in children with CD compared to reference children. In contrast, stimulation with insulin tended to induce high FOXP3 mRNA expression in T1D children compared to reference children (P= 0.057). In conclusion, children with only T1D generally showed a lower FOXP3 mRNA expression than did children with CD, or with T1D in combination with CD, which suggests impaired regulation of the immune system in children with T1D. PMID- 19120313 TI - Autoreactive T cell response in CD25-negative fraction of peripheral blood mononuclear cells in established type 1 diabetes. AB - CD4(+)CD25(+) T cells (Tregs) play a critical role in maintaining dominant peripheral tolerance, and pathogenic autoreactive T cells may be frequent in the CD25-negative fraction of peripheral blood mononuclear cells (PBMCs) from patients with autoimmune disease. We therefore investigated whether T cell autoimmune responses to recombinant GAD65 can be detected by the use of ELISPOT assay in the CD25-negative fraction of PMBCs from Japanese type 1 diabetes (T1D) patients. The frequency of CD4(+)CD25(+) T cells was not different among patients with newly developed T1D, established T1D, and healthy controls. The CD25 positive cell-depleted fraction was obtained by negative selection with antihuman CD25 magnetic beads, reducing the number of CD4(+)CD25(+) T cells from 4-5% to less than 1%. In whole PBMC fraction, there was a significant elevation of IFN gamma spots in PBMCs from recently diagnosed patients with T1D (P < 0.05), whereas the number of IFN-gamma spots from patients with established T1D was not significant. In the CD25-negative fraction, unlike whole PBMCs, we observed the significant IFN-gamma spots to GAD65 in the fraction from patients with established T1D (P < 0.05), but not in those with recently diagnosed disease. The phenomena were not observed for IL-4 spots. Our data suggest a possible role of Tregs maintaining dominant peripheral tolerance in T1D and application of further improved T cell assay detecting autoimmunity even in established T1D. PMID- 19120314 TI - Gene expression profiles stratified according to type 1 diabetes mellitus susceptibility regions. AB - The MHC region (6p21) aggregates the major genes that contribute to susceptibility to type 1 diabetes (T1D). Three additional relevant susceptibility regions mapped on chromosomes 1p13 (PTPN22), 2q33 (CTLA-4), and 11p15 (insulin) have also been described by linkage studies. To evaluate the contribution of these susceptibility regions and the chromosomes that house these regions, we performed a large-scale differential gene expression on lymphomononuclear cells of recently diagnosed T1D patients, pinpointing relevant modulated genes clustered in these regions and their respective chromosomes. A total of 4608 cDNAs from the IMAGE library were spotted onto glass slides using robotic technology. Statistical analysis was carried out using the SAM program, and data regarding gene location and biological function were obtained at the SOURCE, NCBI, and FATIGO programs. Three induced genes were observed spanning around the MHC region (6p21-6p23), and seven modulated genes (5 repressed and 2 repressed) were seen spanning around the 6q21-24 region. Additional modulated genes were observed in and around the 1p13, 2q33, and 11p15 regions. Overall, modulated genes in these regions were primarily associated with cellular metabolism, transcription factors and signaling transduction. The differential gene expression characterization may identify new genes potentially involved with diabetes pathogenesis. PMID- 19120315 TI - Evaluation of cytokine production from peripheral blood mononuclear cells of type 1 diabetic patients. AB - This study aims to evaluate the production of cytokines, tumor necrosis factor (TNF), and interleukin 10 (IL-10) in peripheral blood mononuclear cells (PBMCs) from type 1 diabetic (T1D) patients by means of intracellular staining, flow cytometry, and ELISA and to correlate it with inadequate (IN) and adequate (A) metabolic controls. We studied 28 patients with T1D and 20 healthy individuals (C) paired by sex and age. T1D patients were divided in patients with IN and A metabolic control. PBMC cultures were stimulated with LPS to evaluate TNF or were stimulated with PMA/ionomycin or concanavalin A to evaluate IL-10. The TNF levels in supernatant of stimulated cultures, evaluated by ELISA, of diabetic patients were similar to those of healthy individuals, although the percentage of CD 33(+) cells that were positive for TNF was higher in the T1D IN group compared to the T1D A group (P= 0.01). Similarly, the IL-10 levels evaluated by ELISA in stimulated cultures of T1D patients were not different from those in the control group; moreover, the percentage of CD3(+) cells positive for intracellular IL-10 were higher in the T1D IN group compared to C groups (P= 0.007). The increased levels of cytokines in T1D IN diabetic patients, with reduction in the A group, suggests that hyperglycemia stimulates an inflammatory state that can result in a deficient immune cellular response. The data suggest that assessment by intracellular staining seems to be more accurate than the ELISA technique in evaluating diabetic patients. PMID- 19120316 TI - Insulitis in human type 1 diabetes. AB - Pancreatic tissues were analyzed immunohistologically in patients with autoimmune and fulminant type 1 diabetes (T1D) and control subjects. Both beta and alpha cells were decreased in fulminant T1D, but only beta cells were significantly decreased in autoimmune T1D. Insulitis was seen in both subtypes of T1D, but it remained longer in autoimmune than in fulminant T1D. Lymphocytic infiltration to the exocrine pancreatic tissue was observed only in fulminant T1D, whereas immunologically abnormal findings, such as increased expression of MHC class I molecule and Fas antigen in islet cells and Fas-ligand expression in infiltrating lymphocytes, were detected only in autoimmune T1D. From these findings, together with clinical features, it could be concluded that in autoimmune T1D, beta cells are assumed to be destroyed through a long-standing autoimmune process, whereas in fulminant T1D, beta cells seem to be destroyed very rapidly, probably by a destructive process triggered by viral infection. PMID- 19120318 TI - Induction of apoptosis of Beta cells of the pancreas by advanced glycation end products, important mediators of chronic complications of diabetes mellitus. AB - We herein report cytotoxicity of advanced glycation end-products (AGEs) on pancreatic beta cells. AGEs stimulated reactive oxygen species (ROS) generation but did not arrest proliferation of the INS-1 cell line. Pancreatic beta cell lines or primary cultured islets possess a receptor for AGE (RAGE), and its expression increased after AGE treatment. TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in INS-1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde (AGE2) or glucoaldehyde (AGE3), compared with those conjugated with glucose (AGE1). Reaction of INS-1 cells to Ki67, which is a cellular marker for proliferation, was also increased after AGE treatment. The ability of primary cultured islets to secrete insulin was retained even after AGE treatment under either low or high glucose conditions. The antiserum against RAGE partially prevented AGE-induced cellular events. Treatment of beta cells with the antioxidant metallothionein results in a significant reduction in pathologic changes. AGEs might be able to induce apoptosis as well as proliferation of pancreatic beta cell lines or primary cultured islets. Moreover, antibody array showed that RAD51 and RAD52 were significantly decreased in AGE2-treated INS-1 cells. AGEs might inhibit homologous DNA recombination for repairing DNA of INS-1 cells damaged by ROS generation. It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic beta cells. AGEs might deteriorate function of pancreatic beta cells in patients with long-term hyperglycemia. PMID- 19120317 TI - Immunoregulatory pathways controlling progression of autoimmunity in NOD mice. AB - The activation, expansion, and survival of regulatory T cells (Tregs) as well as the expression of their suppressive capacities result from distinct signaling pathways involving various membrane receptors and cytokines. Multiple studies have shown that thymus-derived naturally occurring Tregs constitutively express the forkhead/winged helix transcription factor FoxP3 in addition to high levels of CD25, the negative co-stimulatory molecule CTLA-4, and the glucocorticoid induced TNF receptor-related protein GITR. At variance, adaptive or induced Tregs acquire these phenotypic markers as they differentiate in the periphery, following adequate stimulation in the appropriate environment, together with their capacity to produce immunomodulatory cytokines (mainly, IL-4, IL-10 and TGF beta) and to display regulatory capacities. However, none of these molecules but FoxP3 are restricted to Tregs since they may also be expressed and upregulated on activated effector T cells. This explains why different hypotheses were proposed to interpret interesting reports showing that in vivo abrogation of CTLA-4 signaling using neutralizing CTLA-4 antibodies triggers different autoimmune or immune-mediated manifestations. Thus, an effect on pathogenic T cell effectors and/or Tregs has been proposed. Here we present and discuss recent results we obtained in the nonobese diabetic (NOD) mouse model of spontaneous autoimmune diabetes, arguing for a key role of CTLA-4 in the functional activity of Tregs. Moreover, data are presented that simultaneous blockade of CTLA4 and TGF-beta further impairs immunoregulatory circuits that control disease progression. PMID- 19120319 TI - Preparation and testing of diabetes autoantibody controls. AB - Stocks of the WHO islet cell antibody, GAD(65) antibody, and IA-2 antibody standard (NIBSC 97/550) are now very limited. We have therefore made and tested a series of control preparations in which human monoclonal autoantibodies to IA-2 and to GAD(65) were diluted in antibody-negative human serum to different concentrations. Three different diabetes autoantibody controls (DAC 1-3) were made as was a negative control preparation. Aliquots containing 1 mL of autoantibodies in serum were freeze-dried. After reconstitution (with 1 mL of water) the controls were tested by (125)I-IA-2 immunoprecipitation assay (IPA), (125)I-GAD(65) IPA, GAD(65) Ab ELISA and IA-2 Ab ELISA (kits from RSR Ltd.) and for ICA by immunofluorescence test (IFT). DAC1 is particularly suitable as a control for the (125)I IA-2 IPA; DAC2 is suitable for the (125)I-GAD(65) IPA, GAD(65) Ab ELISA, and IA-2 Ab ELISA; and DAC3 is suitable for the ICA IFT. Freeze dried preparations showed good stability at 37 degrees C. Reconstituted liquid preparations were stable when stored at 4 degrees C and at 37 degrees C. Availability of an essentially unlimited supply of these reagents should be useful in establishing reproducible and comparable measurements of diabetes autoantibodies in different laboratories using different assays. PMID- 19120320 TI - Tumor necrosis factor alpha inducing spatial interactions between calcium-sensing receptor and L-type voltage-dependent calcium channel. AB - The temporal and spatial regulation of intracellular calcium concentration ([Ca(2+)](i)) is very crucial for maintaining calcium ion homeostasis within cells, and consequently in the regulation of cellular functions such as beta cell replication and differentiation, insulin secretion, and apoptosis. Calcium ion regulatory proteins playing major roles in these processes include L-type voltage dependent calcium channels (L-type VDCCs) and calcium-sensing receptors (CaRs). Tumor necrosis factor alpha (TNF-alpha), a cytokine, is widely known to activate nuclear factor-kappa B (NF-kappaB) transcription in beta cells. Confocal fluorescence imaging data suggest increased co-localization of CaRs with L-type VDCCs upon treatment of beta cells with TNF-alpha, thereby indicating increased membrane-delimited spatial interactions between these two membrane proteins. PMID- 19120321 TI - Tracking down immune markers from alternative system pathway factors in a diabetic population. AB - Hyperglycemia associated with type 1 diabetes (T1D) alters the host immune system, resulting in a predisposition to infectious diseases. The high risk of infection in the diabetic population may lead to life-threatening situations. The early proteins of the alternative complement system pathway, constituting factors P, B, and D, have been shown to play an important role in preventing infection because they form a membrane attack complex (MAC)-C5-9, which debilitates the target microbes and/or molecules via cytotoxic and cytolytic reactions. Patients who are devoid of or contain low levels of these proteins may be susceptible to developing chronic infections. We have observed striking differences in partially fractionated serum proteins in diabetic patients (type 2) relative to controls, through single and two-dimensional gel electrophoresis. Our data, obtained from 50 diabetic patients in the age group of 25-45 years, who had the disease for fewer than 5 years, indicated patterns in low- and high-molecular-weight proteins, which could be grouped into five different categories with minor differences in their respective levels of protein expression. Immunoblot assay could barely detect the presence of properdin expression in diabetic patients. Quantization by ELISA in 99 patients indicated low levels of properdin expression in 70% of 50 diabetic patients (6.5 +/- 3 mug/mL) when compared to nondiabetic controls (19.5 +/- 8.5 mug/mL). This study concluded that patients with low expression of properdin should be advised to take extensive preventive measures and seek early management with appropriate treatments against infection. PMID- 19120322 TI - Ear acupuncture or local anaesthetics as pain relief during postpartum surgical repair: a randomised controlled trial. AB - OBJECTIVE: To evaluate two methods of pain relief during postpartum surgical repair in regard to effectiveness, wound healing and patient evaluation. DESIGN: A randomised controlled trial testing a pragmatic set-up of brief training of clinicians. SETTING: Delivery ward at a Danish district hospital with approximately 1600 annual deliveries. POPULATION: Primiparous women with a vaginal delivery at term who needed surgical repair of lacerations to the labia or the vagina, perineal lacerations of first or second degree or mediolateral episiotomies. METHODS: The trial was set up to evaluate the effect of a brief 2 hour hands-on training in the use of ear acupuncture. All midwives (n = 36) in the department had previous experience in using acupuncture for obstetric pain relief. Pain and wound healing were evaluated using validated scores. Data collection was performed by research assistants blinded towards treatment allocation. Randomisation was computer assisted. A total of 207 women were randomised to receive ear acupuncture (105) and local anaesthetics (102), respectively. MAIN OUTCOME MEASURES: The primary outcome was pain during surgical repair. Secondary outcomes were wound healing at 24-48 hours and 14 days postpartum, participant satisfaction, revision of wound or dyspareunia reported 6 months postpartum. RESULTS: Pain during surgical repair was more frequently reported by participants allocated to ear acupuncture compared with participants receiving local anaesthetics (89 versus 54%, P < 0.01). Pain intensity during surgical repair was also reported higher (Visual Analogue Scale score 3.5 versus 1.5, P < 0.01). The ear acupuncture group received more additional pain relief during repair (53 versus 19%, P < 0.01). No difference was observed in wound healing at 24-48 hours or 14 days postpartum. Revision of wounds was rare, and no difference occurred in this trial. Comparable proportions of participants reported dyspareunia at 6 months. Patient satisfaction with the allocated pain relief method was lower in the ear acupuncture group (69 versus 91%, P < 0.01) and fewer women would recommend the method to a friend (74 versus 91%, P < 0.01). CONCLUSIONS: Ear acupuncture as used in this trial was less effective for pain relief compared with a local anaesthetic. No difference was observed in wound healing, need for revision of wound or dyspareunia. Patient satisfaction with allocated pain-relief method was lower in the ear acupuncture group. PMID- 19120323 TI - Compound heterozygosity for ARS component B mutations in a Dutch patient with mal de Meleda. PMID- 19120324 TI - Pulsed dye laser vs. intense pulsed light for port-wine stains: a randomized side by-side trial with blinded response evaluation. AB - BACKGROUND: Pulsed dye lasers (PDLs) are considered the treatment of choice for port-wine stains (PWS). Studies have suggested broadband intense pulsed light (IPL) to be efficient as well. So far, no studies have directly compared the PDL with IPL in a randomized clinical trial. OBJECTIVES: To compare efficacy and adverse events of PDL and IPL in an intraindividual randomized clinical trial. METHODS: Twenty patients with PWS (face, trunk, extremities; pink, red and purple colours; skin types I-III) received one side-by-side treatment with PDL (V-beam Perfecta, 595 nm, 0.45-1.5 ms; Candela Laser Corporation, Wayland, MA, U.S.A.) and IPL (StarLux, Lux G prototype handpiece, 500-670 and 870-1400 nm, 5-10 ms; Palomar Medical Technologies, Burlington, MA, U.S.A.). Settings depended on the preoperative lesional colour. Treatment outcome was evaluated by blinded, clinical evaluations and by skin reflectance measurements. RESULTS: Both PDL and IPL lightened PWS. Median clinical improvements were significantly better for PDL (65%) than IPL (30%) (P = 0.0004). A higher proportion of patients obtained good or excellent clearance rates with the PDL (75%) compared with IPL (30%) (P = 0.0104). Skin reflectance also documented better results after PDL (33% lightening) than IPL (12% lightening) (P = 0.002). Eighteen of 20 patients preferred to receive continued treatments with PDL (P = 0.0004). No adverse events were observed with PDL or IPL. CONCLUSIONS: Both the specific PDL and IPL types of equipment used in this study lightened PWS and both were safe with no adverse events. However, the PDL conveyed the advantages of better efficacy and higher patient preference. PMID- 19120325 TI - Quality of life and psychological distress in patients with cutaneous lymphoma. AB - BACKGROUND: Cutaneous lymphomas may have a profound impact on patients' health related quality of life (HRQoL) and psychological well-being. OBJECTIVES: To evaluate HRQoL and psychological distress in patients with cutaneous lymphoma, and to evaluate them in relation to personal and clinical characteristics. METHODS: Patients with cutaneous T-cell lymphoma or cutaneous B-cell lymphoma (CBCL) were consecutively recruited in a dermatological hospital. Data on HRQoL were collected using a dermatology-specific questionnaire, the Skindex-29, and an oncology-specific questionnaire, the EORTC QLQ-C30. RESULTS: Of 95 patients, there were 24 with CBCL, 59 with mycosis fungoides (MF) and 12 with Sezary syndrome (SS). The most frequent items reported in Skindex-29 were itching and sensitive skin, being annoyed by the disease, worry that it could get worse, affected interactions, and impairment in sexual life. The most frequent problems appearing from the EORTC QLQ-C30 analysis were fatigue, pain and insomnia. A worse HRQoL was observed for all the scales in patients with SS, followed by MF, and CBCL. HRQoL impairment in all histotypes was higher in women than in men, in patients with probable anxiety or depression, and when the disease worsened. The highest prevalence of probable anxiety or depression was observed in patients treated with systemic steroids (60%) and interferon (50%). CONCLUSIONS: The detailed evaluation of HRQoL and psychological problems in patients with cutaneous lymphomas, and their relationship with clinical variables, may give important information on the burden of the disease for patients, and thus improve communication and satisfaction with care. PMID- 19120326 TI - Suppression of mTOR complex 2-dependent AKT phosphorylation in melanoma cells by combined treatment with rapamycin and LY294002. AB - BACKGROUND: Inhibition of mTOR complex 1 (mTORC1) with rapamycin leads to phosphorylation of AKT in some cancer cells, with unknown biological consequences. The role of this phosphorylation in melanoma is unknown, although preliminary clinical data indicate poor activity of rapalogues in melanoma. OBJECTIVES: We aimed at elucidating the role of AKT phosphorylation after mTORC1 inhibition in melanoma cells. METHODS: Western blotting, apoptosis assays, cell cycle analyses and viability assays were performed to analyse the effects of rapamycin and LY294002 treatment on melanoma cells. For suppression of mTOR complex 2 (mTORC2) an siRNA directed against rictor was used. RESULTS: Rapamycin showed limited effects on cell viability but resulted in strong and lasting AKT phosphorylation in melanoma cells. Combined PI3K/mTOR inhibition with LY294002 had pronounced effects on viability but also led to increased AKT phosphorylation after prolonged treatment. In contrast, combination of rapamycin plus LY294002 suppressed AKT phosphorylation. Suppression of AKT phosphorylation did not correlate with decreases in cell viability. Inhibition of mTORC2 led to reduced levels of phosphorylated AKT. CONCLUSIONS: mTORC1 inhibition with rapamycin and with LY294002 can lead to AKT phosphorylation in melanoma cells via mTORC2. Combination of rapamycin and LY294002 suppresses AKT phosphorylation but without significant effect on treatment efficacy. PMID- 19120327 TI - Refractory urticarial vasculitis responsive to anti-B-cell therapy. PMID- 19120328 TI - Troglitazone suppresses transforming growth factor-beta1-induced collagen type I expression in keloid fibroblasts. AB - BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)-gamma agonists are increasingly used in patients with diabetes and some studies have suggested a beneficial effect on organ fibrosis. However their effects on dermal fibrosis in keloids are unknown. OBJECTIVE: To investigate the effect of the PPAR-gamma agonist troglitazone on transforming growth factor (TGF)-beta1-induced collagen type I expression in keloid fibroblasts. METHODS: Keloid fibroblasts were cultured and exposed to different concentrations of troglitazone in the presence of TGF-beta1. The mRNA expression of PPAR-gamma was determined by semiquantitative reverse transcriptase-polymerase chain reaction. The protein of PPAR-gamma, Smad2, Smad3, phoshpo-Smad2/3 and collagen type I was determined by Western blotting and collagen synthesis was evaluated by measuring (3)H-proline incorporation. The effect of troglitazone on cell viability was evaluated by the colorimetric conversion of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide. RESULTS: PPAR-gamma was expressed at a moderate level in keloid fibroblasts. Troglitazone depressed TGF-beta1-stimulated collagen type I expression and collagen synthesis in keloid fibroblasts in a concentration dependent manner. Moreover, troglitazone inhibited expression and phosphorylation of TGF-beta1-induced Smad2/3. Cell viability was unaffected. These inhibitory effects of troglitazone were reversed by the PPAR-gamma-specific antagonist GW9662. CONCLUSIONS: Our data suggest that PPAR-gamma is present in keloid fibroblasts and PPAR-gamma activation inhibits TGF-beta1-induced collagen type I expression at least in part by decreasing collagen synthesis. PPAR-gamma may be a promising therapeutic target for keloids. PMID- 19120329 TI - Efficacy and safety of tacrolimus ointment 0.1% vs. betamethasone 17-valerate 0.1% in the treatment of chronic paronychia: an unblinded randomized study. AB - BACKGROUND: Recent studies have established the pivotal role of irritants and allergens in development of chronic paronychia and the significant improvement with corticosteroid therapy. OBJECTIVES: The objective of this randomized, unblinded, comparative study was to compare the efficacy of tacrolimus ointment 0.1% vs. betamethasone 17-valerate 0.1% in the treatment of chronic paronychia. METHODS: Forty-five patients with chronic paronychia were randomized 1:1:1 to apply twice daily either betamethasone 17-valerate 0.1% or tacrolimus 0.1% ointment or emollient. Protective measures were counselled to all patients. Treatment duration was 3 weeks and patients were followed for an additional 6 weeks. RESULTS: Eight patients in the betamethasone group were considered as cured, two as improved and four as nonresponders at the end of the treatment period. Thirteen patients in the tacrolimus group were considered as cured and one as improved at the end of the treatment period. Nine patients in the emollient group were considered as stable and six failed to respond. Both betamethasone and tacrolimus groups presented statistically significantly greater cure or improvement rates when compared with the emollient group (P<0.001). CONCLUSIONS: Tacrolimus ointment appears to be a more efficacious agent than betamethasone 17-valerate or placebo for the treatment of chronic paronychia. PMID- 19120330 TI - Care of patients with psoriasis: an audit of U.K. services in secondary care. AB - BACKGROUND: Medical professionals require data about the structure and delivery of dermatological services in primary and secondary care in order to identify and tackle variations in standards and monitor the impact of healthcare reforms. The British Association of Dermatologists (BAD) commissioned an audit of the provision of care for patients with psoriasis. OBJECTIVES: To assess the staffing and facilities in dermatology units in the U.K. with a focus on the provision of care for patients with psoriasis. METHODS: Data were collected from 100 dermatology units in the U.K. for 1 year using a questionnaire and a web-based collection system. RESULTS: Key results are as follows. Eighteen per cent (18/98) of units had fewer than 2.0 whole-time equivalent consultants and 20% had no specialist dermatology nurse. Only 23% of units collected diagnostic data on outpatients, and half were unable to supply details about the number of attendances for psoriasis. Seventy-seven units reported admitting patients to dedicated dermatology beds, general medical beds, or both; three-quarters of units had access to dedicated adult dermatology beds. Pharmacy services were not always available for dermatology patients. Only 21 units (21%) had dedicated clinics for patients with psoriasis and 56% of units lacked a clinical psychology service willing to accept adult dermatology patients; 59% (55/93) lacked psychological services for children. Fifty-five per cent had no systemic drug monitoring clinic. Phototherapy was run by dermatology nurses in 93% (88/95) of the units and by physiotherapists in 11% (10/94). Biologics for psoriasis were prescribed in 75% (73/97) of units and in 88% (64/73) of these the BAD guidelines for the use of biologics were known to be followed. Of the seventy-three units prescribing biologic therapies, 64% had a nurse trained in the assessment and administration of biologics, 71% had facilities for outpatient infusions (e.g. for infliximab) and 39% were restricted in prescribing biologic agents because of financial constraints. A quality-of-life score was either inadequately or never recorded in outpatient records in 81% of units, increasing to 88% for inpatient records. The Psoriasis Area and Severity Index score was inadequately or never recorded in 79% of outpatient records and 82% of inpatient records. CONCLUSIONS: Units varied in their capacity to meet BAD guidelines and standards. Among the most significant deficiencies identified were a shortage of specialist dermatology nurses, treatment delivery by untrained nurses and financial constraints on the prescription of biologics for psoriasis. Gaps in data collection and record keeping jeopardize efforts to improve standards of care. PMID- 19120331 TI - Dermoscopy of cutaneous leishmaniasis. AB - BACKGROUND: Dermoscopy has been proposed as a diagnostic tool in the case of skin infections and parasitosis but no specific dermoscopic criteria have been described for cutaneous leishmaniasis (CL). OBJECTIVES: To describe the dermoscopic features of CL. METHODS: Dermoscopic examination (using the DermLite Foto; 3Gen, LLC, Dana Point, CA, U.S.A.) of 26 CL lesions was performed to evaluate specific dermoscopic criteria. RESULTS: We observed the following dermoscopic features: generalized erythema (100%), 'yellow tears' (53%), hyperkeratosis (50%), central erosion/ulceration (46%), erosion/ulceration associated with hyperkeratosis (38%) and 'white starburst-like pattern' (38%). Interestingly, at least one vascular structure described in skin neoplasms was observed in all cases: comma-shaped vessels (73%), linear irregular vessels (57%), dotted vessels (53%), polymorphous/atypical vessels (26%), hairpin vessels (19%), arborizing telangiectasia (11%), corkscrew vessels (7%) and glomerular like vessels (7%). Combination of two or more different types of vascular structures was present in 23 of 26 CL lesions (88%), with a combination of two vascular structures in 13 cases (50%) and three or more in 10 cases (38%). CONCLUSIONS: Characteristic dermoscopic structures have been identified in CL. Important vascular patterns seen in melanocytic and nonmelanocytic tumours are frequently observed in this infection. PMID- 19120332 TI - Chronic urticaria: do urticaria nonexperts implement treatment guidelines? A survey of adherence to published guidelines by nonexperts. AB - BACKGROUND: Guidelines including level of evidence and grade of recommendation were recently published for chronic urticaria (CU). OBJECTIVES: To describe the therapeutic approach in patients with CU, and to depict how recent guidelines are implemented in the daily practice of management of CU. METHODS: We performed a cross-sectional multicentre study through a questionnaire answered by 139 specialists. In total, 695 patients were evaluated, mean+/-SD age 42.3+/-15 years, 62.1% women. Of the patients, 168 were treated by an allergist, 473 by a dermatologist and in 54 cases the specialist was not stated. The drug prescribed was the main variable, and chi2 and Fisher's tests were utilized for the statistical analysis. RESULTS: Nonsedating anti-H1 antihistamines taken regularly were the most common drugs prescribed, followed by nonsedating anti-H1 antihistamines taken as needed, corticosteroids, sedating antihistamines taken regularly, sedating antihistamines taken as needed, anti-H2 antihistamines, leukotriene antagonists, ciclosporin and doxepin. Nonsedating antihistamines plus corticosteroids was the most frequent drug combination prescribed. When comparing between allergists and dermatologists we found a positive and significant correlation only between prescription of cetirizine, dexchlorfeniramine, leukotriene antagonists and anti-H2 antihistamines and being treated by an allergist. A positive correlation was found with desloratadine and being seen by a dermatologist. We did not find any difference in CU management in the rest of the treatments studied. CONCLUSIONS: It is surprising that a large amount of sedating antihistamines was prescribed. In many instances these were prescribed as needed. This fact could have a negative impact on urticaria control and patient satisfaction. It seems difficult for the nonexpert to differentiate between CU and any kind of physical urticaria. PMID- 19120333 TI - Medium-dose ultraviolet (UV) A1 vs. narrowband UVB phototherapy in atopic eczema: a randomized crossover study. AB - BACKGROUND: Ultraviolet (UV) A1 and narrowband (NB)-UVB have been reported to be effective treatments for atopic eczema (AE). OBJECTIVES: We aimed to compare the efficacy of medium-dose UVA1 and NB-UVB mono-phototherapy in patients with AE. METHODS: A randomized double-blind controlled crossover trial (ClinicalTrials.gov Identifier: NCT00419406) was conducted in which patients with AE received a 6 week course of both medium-dose UVA1 and NB-UVB. Clinical efficacy was assessed using the Six Area, Six Sign, Atopic Dermatitis (SASSAD) score and a visual analogue scale for pruritus. Assessment of health-related quality of life was performed using the Skindex-29. Total immunoglobulin E (IgE) and eosinophilic cationic protein (ECP) were evaluated at baseline and after each phototherapy course. RESULTS: Twenty-eight patients who completed both UVA1 and NB-UVB phototherapy courses on an intention-to-treat basis were analysed according to the crossover design. Both interventions were associated with significant clinical improvement but there was no significant difference between treatments with respect to the mean +/- SD relative reduction (RR) of the clinical scores (SASSAD, 43.7 +/- 31.4% vs. 39.4 +/- 24.1%, P = 0.5; pruritus score, 16 +/- 61.8% vs. 25.2 +/- 30.5%, P = 0.5, respectively). There was no significant difference in the mean +/- SD RR of the Skindex-29 after UVA1 and NB-UVB phototherapy (12.7 +/- 18.8% vs. 16.5 +/- 17.6%, P = 0.1). Changes in the total IgE and ECP levels following UVA1 and NB-UVB did not differ significantly (P = 0.3 and P = 0.9, respectively). CONCLUSIONS: A 6-week course of NB-UVB and UVA1 phototherapy of AE resulted in significant clinical improvement. With regard to efficacy and tolerability, both phototherapeutic modalities may be considered comparably good. PMID- 19120334 TI - Morphological and genetic analysis of steatocystoma multiplex in an Asian family with pachyonychia congenita type 2 harbouring a KRT17 missense mutation. PMID- 19120335 TI - Metastatic prostate cancer presenting as paraneoplastic pemphigus: a favourable clinical response to combined androgen blockade and conventional immunosuppressive therapy. PMID- 19120336 TI - Enhancement of bioavailability by lowering of fat content in topical formulations. AB - BACKGROUND: The cosmetic properties of topical formulations are important parameters for the adherence to treatment, where modern oil-in-water emulsions are considered more acceptable compared with ointments. After application of an emulsion to the skin, the concentration of active ingredients in the formulation residue on the skin will increase, due to evaporation of volatile ingredients. OBJECTIVES: The aim of the present study was to investigate the effect of changes in vehicle fatty content on the skin penetration of two active ingredients: benzyl nicotinate (BN) and betamethasone valerate (BV). METHODS: Formulations containing 0.5% BN and 0.3% BV in vehicles with different lipid content (10-80%) were applied in a randomized and double-blind manner to the forearm of healthy volunteers. The changes in skin colour (erythema and blanching) were then monitored visually and with a new noninvasive instrument. RESULTS: The BN formulation containing 10% fat induced erythema more rapidly and with higher intensity than the formulations with higher fat content. Increased efficacy was also observed from the low-fat content formulation of BV, which gave more blanching than the formulations with high fat content. CONCLUSIONS: The rate of penetration of the active ingredients was inversely related to the lipid content, i.e. simple changes of the cosmetic properties by modifications of the lipid content may affect the efficacy of a formulation. PMID- 19120337 TI - First experience with enteric-coated mycophenolate sodium (Myfortic) in severe recalcitrant adult atopic dermatitis: an open label study. AB - BACKGROUND: Severe atopic dermatitis (AD) is often treated successfully with oral immunosuppressive drugs such as cyclosporin (CsA) or oral corticosteroids. However, some patients develop adverse effects or are unresponsive to these first choice oral immunosuppressive drugs. OBJECTIVES: To evaluate whether enteric coated mycophenolate sodium (EC-MPS) is an effective treatment in patients with severe, recalcitrant AD. METHODS: Ten patients with severe, recalcitrant AD were treated with EC-MPS 720 mg twice daily for 6 months. All patients had to discontinue other oral immunosuppressive drugs due to adverse effects (n = 8) or nonresponsiveness (n = 2). Disease activity was monitored using the Severity Scoring of Atopic Dermatitis (modified SCORAD) index and the Leicester Sign Score (LSS). Additionally, the level of serum thymus and activation-regulated cytokine (TARC) was measured. During treatment, safety laboratory examination was performed. Total serum immunoglobulin E (IgE) was followed during treatment. Use of topical corticosteroids was recorded before and during treatment. RESULTS: Compared with baseline, the mean scores for disease activity significantly decreased during treatment with EC-MPS [modified SCORAD (P = 0.04), LSS severity (P = 0.01), LSS extent (P = 0.01)]. In addition, serum TARC levels and total serum IgE levels significantly decreased after treatment compared with before (P = 0.03; P = 0.05). Disease activity decreased after approximately 2 months of treatment and stabilized during the 6-month treatment period. No differences in the amount of topical corticosteroids used in the 6 months prior to treatment compared with the 6-month treatment period were found (P = 0.4). None of the patients discontinued use of EC-MPS and only mild adverse effects were seen. CONCLUSIONS: In this study EC-MPS at a dose of 720 mg twice daily for 6 months has proven to be an effective and well-tolerated treatment for patients with severe, recalcitrant AD. PMID- 19120338 TI - Autosomal dominant junctional epidermolysis bullosa. AB - BACKGROUND: Epidermolysis bullosa (EB) encompasses a heterogeneous group of inherited skin disorders associated with trauma-induced blistering. The junctional forms of EB (JEB), Herlitz JEB, non-Herlitz JEB and JEB associated with pyloric atresia have all been attributed to autosomal recessive inheritance. We describe a 7-year-old girl with defective dental enamel, trauma-induced blistering and subsequent scarring. Her mother, a carrier of the mutation p.G627V in the collagen XVII gene (COL17A1) had evidence of hypoplastic dental enamel without skin blistering. Her grandmother had non-Herlitz JEB as a result of a compound heterozygous mutation in COL17A1 (p.G627V and c.3514ins25). OBJECTIVES: To explore the molecular, ultrastructural and immunofluorescence findings of the first case of dominant JEB. METHODS: Mutational analysis of COL17A1 was performed on the proband's genomic DNA. In addition, transmission electron microscopy and immunofluorescence microscopy were performed on a nonlesional skin biopsy from the proband and an unrelated healthy control. RESULTS: Direct sequencing revealed a heterozygous glycine substitution mutation, p.G627V, in COL17A1. No discernible morphological abnormalities were found on transmission electron microscopy; however, immunofluorescence microscopy revealed findings of an altered distribution pattern for type XVII collagen epitopes close to the dermal epidermal junction. CONCLUSION: This report describes the first case of dominant JEB. Although some heterozygous mutations in COL17A1 are known to cause dental abnormalities none were associated with skin fragility. The dominant-negative interference between the proband's mutated type XVII collagen and the wild-type allele appears to render the skin prone to trauma-induced blister formation. Alternatively, other undisclosed modifying genetic or epigenetic factors might explain why the patient gets blistering whereas her mother, who has the same COL17A1 mutation, has no skin fragility. PMID- 19120339 TI - Kindler syndrome: a focal adhesion genodermatosis. AB - Kindler syndrome (OMIM 173650) is an autosomal recessive genodermatosis characterized by trauma-induced blistering, poikiloderma, skin atrophy, mucosal inflammation and varying degrees of photosensitivity. Although Kindler syndrome is classified as a subtype of epidermolysis bullosa, it has distinct clinicopathological and molecular abnormalities. The molecular pathology of Kindler syndrome involves loss-of-function mutations in a newly recognized actin cytoskeleton-associated protein, now known as fermitin family homologue 1, encoded by the gene FERMT1. This protein mediates anchorage between the actin cytoskeleton and the extracellular matrix via focal adhesions, and thus the structural pathology differs from other forms of epidermolysis bullosa in which there is a disruption of the keratin intermediate filament-hemidesmosome network and the extracellular matrix. In the skin, fermitin family homologue 1 is mainly expressed in basal keratinocytes and binds to the cytoplasmic tails of beta1 and beta3 integrins as well as to fermitin family homologue 2 and filamin-binding LIM protein 1. It also plays a crucial role in keratinocyte migration, proliferation and adhesion. In this report, we review the clinical, cellular and molecular pathology of Kindler syndrome and discuss the role of fermitin family homologue 1 in keratinocyte biology. PMID- 19120340 TI - Negative predictive value of drug skin tests in investigating cutaneous adverse drug reactions. AB - BACKGROUND: Drug skin tests are useful in aetiological analyses of cutaneous adverse drug reactions to determine if the drug can be rechallenged, or to avoid a cross-reaction with a substitute drug. OBJECTIVES: To evaluate the negative predictive value of drug skin tests. METHODS: We retrospectively analysed the files of patients referred for drug reactions. We have enrolled those having strictly determined drug reactions with clinical features, delayed onset after drug intake, drug causality assessment, and negative drug skin tests followed by drug administration. Oral provocation tests or substitution tests with a drug of the same class as that suspected of causing the drug reactions were performed. RESULTS: From 1957 files analysed, 200 patients were included. After 403 patch tests, 403 prick tests and 304 intradermal tests, which were all negative, 260 oral provocation tests and 143 substitution tests were done; 307 different drugs were rechallenged. There were 42 positive drug re-administrations in 27 oral provocation tests and 15 substitution tests. The negative predictive value of our drug skin tests was 89.6%. The negative predictive value for beta-lactams was 87% for oral provocation tests and 96% for substitution tests, and for corticosteroids it was 100% and 74%, respectively. CONCLUSIONS: Negative drug skin tests do not eliminate the responsibility of a drug in drug reactions, and must be followed by drug re-administration under hospital surveillance. PMID- 19120341 TI - Human immunodeficiency virus atropy induces modification of subcutaneous adipose tissue architecture: in vivo visualization by high-resolution magnetic resonance imaging. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection generally induces lipodystrophy. For targeted treatment a better understanding of its development is necessary. The utility of high-resolution magnetic resonance imaging (MRI) is explored. OBJECTIVES: The present study presents a way to visualize the adipose tissue architecture in vivo and to inspect modifications associated with the atrophy. METHODS: High-resolution MRI scans with surface coils were performed on the calf and at the lumbar region of three groups of patients: HIV patients with lipoatrophy, HIV patients without lipoatrophy and healthy volunteers. All patients underwent a clinical examination. In addition, dual energy X-ray absorptiometry (DEXA) measurements were taken. On the MRI scans adipose tissue thickness and adipose nodule size were measured. Results High-resolution MRI enabled identification of a clear disorganization of adipose tissue in patients with lipoatrophy. In addition, these patients presented a very small adipose tissue thickness on the calf and a very small nodule size. RESULTS: led to the hypothesis that adipose tissue disorganization appears before changes in DEXA measurements or clinically visible modifications. CONCLUSIONS: High-resolution MRI enabled visualization in vivo of precise changes in tissue organization due to HIV lipoatrophy. This imaging technique should be very informative for better monitoring of the atrophy. PMID- 19120342 TI - The ultrastructure of acantholysis in pemphigus vulgaris. PMID- 19120343 TI - Characterization of angiotensin-converting enzyme expression during epidermis morphogenesis in humans: a potential marker for epidermal stem cells. AB - BACKGROUND: Recent evidence has revealed that angiotensin-converting enzyme (ACE) participates in cutaneous wound healing and contributes to the pathophysiological process of some skin diseases. However, little is known about the role of ACE in epidermis morphogenesis during development. OBJECTIVE: To clarify the expression pattern of ACE during embryonic development of human skin. METHODS: Skin samples were obtained from aborted fetuses at different gestational ages and from healthy individuals. Localization of ACE, together with beta(1)-integrin, keratin 19 (K19) and p63 was examined by immunofluorescence and immunohistochemical staining. RESULTS: In human fetal skin, at 11-13 weeks of gestation, ACE-positive cells were observed in the primitive epidermis. As the fetuses developed, ACE positive cells appeared in all the epidermal layers. From 21 weeks of gestation, ACE expression was largely restricted to the basal layer of the fetal epidermis. In contrast, ACE-positive cells were found only in the adult skin basal layer which harbours epidermal stem cells. To explore the possible link between ACE and epidermal stem cells, we further examined the expression of beta(1)-integrin, K19 and p63, the putative markers for epidermal stem cells. Consistent with the results of ACE expression, from 21 weeks of gestation, the expression of beta(1) integrin, K19 and p63 was mainly confined to the basal layer. Immunofluorescent double labelling revealed that ACE-positive cells substantially overlapped with beta(1)-integrin-, K19- and p63-positive cells. CONCLUSIONS: Our results suggest that ACE may play a role in human epidermis morphogenesis during fetal life and serve as an unrecognized marker for keratinocyte progenitor cells. PMID- 19120344 TI - The CYP26 inhibitor R115866 potentiates the effects of all-trans retinoic acid on cultured human epidermal keratinocytes. AB - BACKGROUND: All-trans retinoic acid (RA) is known to regulate keratinocyte proliferation and differentiation, and retinoids are used as therapeutic agents in certain dermatological disorders, such as psoriasis and acne. Epidermal expression of the heparin-binding epidermal growth factor-like growth factor (HB EGF) is induced by RA treatment and HB-EGF is responsible for RA-mediated epidermal hyperplasia in vivo. RA also induces HB-EGF expression in cultured keratinocytes and alters their differentiating phenotype. R115866 is a specific inhibitor of the cytochrome P450 isoform CYP26, which is involved in the metabolic inactivation pathway of RA. Thereby, R115866 is thought to be able to increase the intracellular levels of endogenous RA. OBJECTIVES: To determine whether or not R115866 potentiates the effect of low concentrations of RA on keratinocytes. METHODS: We analysed HB-EGF, involucrin and keratin 10 mRNA and protein levels in autocrine human keratinocyte cultures incubated for 18 h with RA or R115866 alone and with RA and R115866 combinations. RESULTS: RA induced HB EGF and involucrin expression in a concentration-dependent manner, whereas it inhibited keratin 10 expression. R115866 alone had no effect on the expression of these genes. However, when R115866 was combined with low concentrations of RA, HB EGF and involucrin expression was induced. CONCLUSION: These results strongly suggest that R115866 potentiates the effects of RA on epidermal keratinocytes when RA is present at low concentrations. PMID- 19120345 TI - CCL18 is expressed in patients with bullous pemphigoid and parallels disease course. AB - BACKGROUND: The autoimmune skin disease bullous pemphigoid (BP) is characterized by subepidermal blister formation and a strong dermal infiltrate of mononuclear cells and eosinophils as well as a T-helper (Th) 2-dominated cytokine milieu. CCL18 is a chemokine, with unknown receptor counterpart, frequently associated with inflammatory Th2-type responses. OBJECTIVES: The study was performed to investigate an association of CCL18 with BP. METHODS: CCL18 was determined by enzyme-linked immunosorbent assay in serum and blister fluid of patients with BP, pemphigus vulgaris and healthy individuals. In vitro chemotaxis assays were performed to demonstrate migration of peripheral blood mononuclear cells to BP blister fluid. Immunohistology and immunofluorescence staining were used to evaluate CCL18 expression in skin. RESULTS: We have found that the levels of CCL18 in sera from patients with BP are 84% higher than those normally observed in healthy individuals. In addition, blister fluid of patients with BP is extremely rich in CCL18, reaching concentrations which are fivefold and sevenfold higher than those found in the sera of patients with BP and healthy individuals, respectively. Using immunofluorescence techniques we identified Langerhans cells, antigen-presenting cells of the dermis and eosinophils as producers of CCL18 in BP skin. We studied the possibility of using CCL18 expression as a biomarker linked to BP by monitoring the serum levels of CCL18 and the disease course of nine patients with BP over a maximum period of 54 months. In this study, CCL18 levels correlated with the disease course in most of the patients. CONCLUSIONS: Our data implicate CCL18 as a functionally relevant chemokine in BP, mediating recruitment of blood mononuclear cells into the hallmark infiltrated skin lesion. The high correlation of CCL18 expression and BP disease suggests that blood levels of this chemokine can be used as an easy method to monitor disease progression and/or efficacy of therapeutic interventions. PMID- 19120346 TI - Economic evaluation of systemic therapies for moderate to severe psoriasis. AB - BACKGROUND: New biologics have dramatically changed therapeutic options for psoriasis, albeit at additional cost. OBJECTIVES: To determine the cost effectiveness and optimal treatment sequence for moderate to severe psoriasis. METHODS: Psoriasis Area and Severity Index (PASI) response rates from 22 randomized controlled trials evaluating biologic (adalimumab, efalizumab, etanercept, infliximab) and nonbiologic systemic (methotrexate, ciclosporin) agents were considered. Short-term efficacy was based on relative probabilities of achieving PASI response (50/75/90) in a meta-analysis of trials. Published evidence and assumptions were used to predict long-term efficacy. Treatment benefits were determined by the relationship between PASI response and the EuroQOL 5D health utility measure. Costs included therapy, administration, monitoring and hospitalization. Incremental cost-effectiveness ratios (ICERs) were calculated and treatments ranked relative to supportive care. RESULTS: Infliximab provided the most incremental quality-adjusted life-years (QALYs) vs. supportive care (0.18 QALYs; 95% confidence interval, CI 0.13-0.24), followed by adalimumab (0.16 QALYs; 95% CI 0.11-0.22). Methotrexate and ciclosporin were less beneficial (0.13 and 0.08 QALYs, respectively) but were cost saving and considered the first two treatments in the optimal sequence. Comparing biologics, adalimumab was most cost effective (ICER pound30 000 per QALY), followed by etanercept ( pound37 000 per QALY), efalizumab ( pound40 000 per QALY) and infliximab ( pound42 000 per QALY). CONCLUSIONS: Methotrexate and ciclosporin are cost effective but require monitoring for toxicities. Of the biologics, adalimumab was most cost effective following conventional systemic treatment failure or inadequate response. Payers and policymakers will have to decide how to utilize their budgets effectively for treating patients with moderate to severe psoriasis. PMID- 19120347 TI - Multiple cytokine-producing peripheral T-cell lymphoma. PMID- 19120348 TI - AL amyloidosis and progression to multiple myeloma with gain(1q). PMID- 19120349 TI - Methylation of tumour suppressor gene promoters in the presence and absence of transcriptional silencing in high hyperdiploid acute lymphoblastic leukaemia. AB - Promoter methylation is a common phenomenon in tumours, including haematological malignancies. In the present study, we investigated 36 cases of high hyperdiploid (>50 chromosomes) acute lymphoblastic leukaemia (ALL) with methylation-specific multiplex ligase-dependent probe amplification to determine the extent of aberrant methylation in this subgroup. The analysis, which comprised the promoters of 35 known tumour suppressor genes, showed that 16 genes displayed abnormal methylation in at least one case each. The highest number of methylated gene promoters seen in a single case was thirteen, with all but one case displaying methylation for at least one gene. The most common targets were ESR1 (29/36 cases; 81%), CADM1 (IGSF4, TSLC1; 25/36 cases; 69%), FHIT (24/36 cases; 67%) and RARB (22/36 cases; 61%). Interestingly, quantitative reverse transcription-polymerase chain reaction showed that although methylation of the CADM1 and RARB promoters resulted in the expected pattern of downregulation of the respective genes, no difference could be detected in FHIT expression between methylation-positive and -negative cases. Furthermore, TIMP3 was not expressed regardless of methylation status, showing that aberrant methylation does not always lead to gene expression changes. Taken together, our findings suggest that aberrant methylation of tumour suppressor gene promoters is a common phenomenon in high hyperdiploid ALL. PMID- 19120350 TI - Prevalence and clinical correlates of JAK2 mutations in Down syndrome acute lymphoblastic leukaemia. AB - Recurrent, prognostically significant chromosomal abnormalities occur in approximately 75% of paediatric acute lymphoblastic leukaemia (ALL), but only infrequently in children with Down syndrome (DS) and ALL. Recently, novel somatic activating mutations in the gene Janus kinase 2 (JAK2) were reported in 18% of DS ALL. Here we report identification and clinical correlates of JAK2 mutations in an independent cohort. JAK2 activating mutations occurred in 10/53 DS ALL cases (18.9%). Mutations were overrepresented in males (P < 0.03), occurred once in association with high hyperdiploidy and were not significantly correlated with age, initial white blood count, or event-free survival. Our results confirm the significance of JAK-STAT pathway activation in DS ALL. PMID- 19120351 TI - Immunoglobulin isotypes in multiple myeloma: laboratory correlates and prognostic implications in total therapy protocols. PMID- 19120352 TI - Weekly imatinib dosage for chronic eosinophilic leukaemia expressing FIP1L1 PDGFRA fusion transcript: extended follow-up. PMID- 19120353 TI - Regulation of iron metabolism through GDF15 and hepcidin in pyruvate kinase deficiency. AB - Iron absorption is inadequately increased in patients with chronic haemolytic anaemia, which is commonly complicated by iron overload. Growth differentiation factor 15 (GDF15) has been identified as a bone marrow-derived factor that abrogates hepcidin-mediated protection from iron overload under conditions of increased erythropoiesis. Increased concentrations of GDF15 have been reported in beta-thalassaemia patients and GDF15 has been found to suppress hepcidin expression in vitro. To further study the interdependencies of iron metabolism and erythropoiesis in vivo, the concentrations of hepcidin and GDF15 were determined in sera from 22 patients with pyruvate kinase deficiency (PKD) and 21 healthy control subjects. In PKD patients, serum hepcidin levels were 13-fold lower than in controls (2.0 ng/ml vs. 26.2 ng/ml) and GDF15 was significantly higher (859 pg/ml vs. 528 pg/ml). Serum hepcidin concentrations correlated positively with haemoglobin and negatively with serum GDF15. These results suggest that GDF15 contributes to low hepcidin expression and iron loading in PKD. PMID- 19120355 TI - Calcium oxalate crystals in the bone marrow. PMID- 19120354 TI - Ethnic differences in F cell levels in Jamaica: a potential tool for identifying new genetic loci controlling fetal haemoglobin. AB - High levels of fetal haemoglobin (HbF) are protective in beta haemoglobinopathies. The proportion of erythrocytes containing HbF (F-cells, FC) was measured in healthy adults of African and Caucasian ancestry to assess the feasibility of localizing genes for the FC trait using admixture mapping. Participants were Afro-Caribbean (AC) blood donors and residents of a rural enclave with a history of recent German admixture (Afro-German, AG) recruited in Jamaica, and Caucasian Europeans recruited in Jamaica and the UK. FC levels were significantly different between groups (P < 0.001); the geometric mean FC level in the AC sample (n = 176) was 3.75% [95% confidence interval (CI) 3.36-4.18], AG sample (n = 631) was 2.77% (95% CI 2.63-2.92), and among Caucasians (n = 1099) was 3.26% (95% CI 3.13-3.39). After adjustment for age, sex, haemoglobin electrophoresis pattern, and HBG2 genotype, FC levels in the AC group remained significantly different (P < 0.001) from those in the Caucasian and the AG group but the difference between the Caucasian and AG groups became non-significant (P = 0.46) despite substantial differences in average ancestry. The data confirm ethnic differences in FC levels and indicate the potential usefulness of these populations for admixture mapping of genes for FC levels. PMID- 19120356 TI - Circulating endothelial cells (CECs) and progenitors (CEPs) in severe haemophiliacs with different clinical phenotype. PMID- 19120357 TI - Inappropriate antidiuretic hormone production by leukaemic blasts in acute myeloid leukaemia. PMID- 19120359 TI - Clinical implications of ELA2-, HAX1-, and G-CSF-receptor (CSF3R) mutations in severe congenital neutropenia. AB - Congenital Neutropenia (CN) is a heterogeneous bone marrow failure syndrome characterized by a maturation arrest of myelopoiesis at the level of the promyelocyte/myelocyte stage with peripheral blood absolute neutrophil counts below 0.5 x 10(9)/l. There are two major subtypes of CN as judged by inheritance: an autosomal dominant subtype, e.g. defined by neutrophil elastase mutations (approximately 60% of patients) and an autosomal recessive subtype (approximately 30% of patients), both presenting with the same clinical and morphological phenotype. Different mutations have been described (e.g. HAX1, p14 etc) in autosomal recessive CN, with HAX1 mutations in the majority of these patients. CN in common is considered as a preleukemic syndrome, since the cumulative incidence for leukemia is more than 25% after 20 years of observation. Leukemias occur in both, the autosomal dominant and recessive subtypes of CN. The individual risk for each genetic subtype needs to be further evaluated. Numbers of patients tested for the underlying genetic defect are still limited. Acquired G-CSFR (CSF3R) mutations are detected in approximately 80% of CN patients who developed acute myeloid leukemia independent of the ELA2 or HAX1 genetic subtype, suggesting that these mutations are involved in leukemogenesis. As the majority of patients benefit from G-CSF administration, HSCT should be restricted to non responders and patients with leukaemic transformation. PMID- 19120358 TI - Expansion of haematopoietic stem cells from normal donors and bone marrow failure patients by recombinant hoxb4. AB - In this study six versions of recombinant human hoxb4 proteins were produced and their effectiveness evaluated in expanding human haematopoietic stem and progenitor cells in vitro and in vivo. An N-terminal-tat and C-terminal histidine tagged version of hoxb4 (T-hoxb4-H) showed the highest activity in expanding colony forming cells (CFCs) and long-term culture-initiating cells (LTC-ICs) when used at 50 nmol/l concentration in cell culture. Human cord blood CD34(+) cells cultured with 50 nmol/l T-hoxb4-H showed a significant increase in severe combined immunodeficient mouse-repopulating cells (SRCs). In a mouse model of immune-mediated bone marrow (BM) failure, T-hoxb4-H showed an additive effect with cyclosporine in alleviating pancytopenia. In addition, T-hoxb4-H expanded CFC and LTC-IC on BM samples from patients with refractory severe aplastic anaemia and myelodysplastic syndromes: after culturing with 50 nmol/l T-hoxb4-H for 4 d, BM cells from 10 of the 11 patients showed increases in CFC and LTC-IC, and the increase in LTC-IC was statistically significant in samples from four patients. Recombinant human hoxb4 could be a promising therapeutic agent for BM failure. PMID- 19120361 TI - Lenalidomide in renal insufficiency--balancing the risks and benefits. PMID- 19120362 TI - Could cytogenetics and minimal residual disease replace conventional risk criteria in adults with Ph-negative acute lymphoblastic leukaemia? PMID- 19120363 TI - From a medical mistake to a clinical warning: the case of HBV mutant virus reactivation in haematological patients. PMID- 19120364 TI - Is rituximab effective in acquired von Willebrand syndrome? PMID- 19120365 TI - Angiogenic mediators of the angiopoietin system are highly expressed by CD10 positive lymphoma cells in angioimmunoblastic T-cell lymphoma. AB - Angioimmunoblastic T-cell lymphoma (AILT) is a malignant disease of peripheral T cell origin that is characterized by a prominent proliferation of high endothelial venules in the lymph node. To investigate angiogenic mechanisms in AILT we measured the angiogenic mediator gene expression levels in the lymph nodes of 54 non-Hodgkin lymphoma patients, by immunostaining and quantitative reverse transcription polymerase chain reaction. Angiogenic mediators angiopoietin (Ang) 1 (ANGPT1), Ang2 (ANGPT2) and their receptor, Tie2 (TEK), vascular endothelial growth factor (VEGF; VEGFA) and its receptor, VEGFR2 (KDR), and hepatocyte growth factor (HGF) and its receptor, c-Met (MET) were all more highly expressed in AILT lymph nodes (16 cases) than in B-cell lymphomas (24 cases). Moreover, significantly higher Ang1 and Tie2 expression was detected in AILT cases with CD10-positive neoplastic T-cells by comparison with unspecified peripheral T-cell lymphoma (14 cases). Immunostaining confirmed the expression of Ang1 and VEGF by both neoplastic T-cells and follicular dendritic cells. These results suggest that the angiopoietin system may play an important role in the development of high vascularity in AILT lymph nodes. Consequently, as neoplastic T-cells and follicular dendritic cells are both increased in AILT and may represent an important source of angiogenic mediators, targeting these cells with anti-angiogenic strategies might represent a novel therapy for AILT. PMID- 19120366 TI - Age-associated difference in gene expression of paediatric acute myelomonocytic lineage leukaemia (FAB M4 and M5 subtypes) and its correlation with prognosis. AB - Acute myeloid leukaemia, French-American-British M4 and M5 subtypes (AML-M4/M5) is frequently associated with MLL gene rearrangement and its incidence is relatively high among infants. Clinically, paediatric AML-M4/M5 has been considered as an intermediate or undefined prognostic group. In this study, we analysed gene expression of 40 paediatric AML-M4/M5 patients excluding inv(16) and t(8;21) patients, and found striking differences among the patients in an age associated manner. In particular, most of the infants displayed very distinct gene expression. On the basis of this difference, we divided paediatric patients into three subgroups (A, B and C) with the average age of 0.3, 3.1 and 6.6 years old respectively. All subgroups included patients with MLL gene rearrangement as well as normal and other karyotypes. Surprisingly, gene expression signatures of MLL gene rearrangement differed substantially among these subgroups. In addition, subgroup C presented extremely poor outcome (3-year event-free survival 28%) whilst eight patients with MLL gene rearrangement in subgroup C had all relapsed within 18 months. These results suggest that age is an important factor contributing to the biology of AML-M4/M5 and the sub-grouping procedures developed in this study could be a powerful tool to identify unfavourable risk patients within paediatric AML-M4/M5. PMID- 19120367 TI - Peripheral blood progenitor cell collection adverse events for childhood allogeneic donors: variables related to the collection and safety profile. AB - The use of children as haematopoietic stem cell donors represents an ethical dilemma. For some investigators, this is even more questionable if the children underwent peripheral blood progenitor cell (PBPC) collection. Due to these controversies, information related to adverse events (AE) in paediatric donors and variables related to the collection is scanty. We analysed 152 PBPC collections, and compared the complications and results between young children, older children and adults. The pattern of AE was shown to vary according to the age of the donor. Older paediatric donors and adults had a higher incidence of complaints related to PBPC priming (54.3% vs. 79.7%, respectively) than the youngest children (12%). On the other hand, these donors had a lower incidence of AE during PBPC collection (19.6% older children, 37.3% adults) mainly related to hypocalcaemia, than the youngest donors, who suffered mainly cardiovascular complications due to hypovolaemia (51.7%). The only variables related to collected cell dose were total blood volume processed per donor body weight, and CD34(+) cell count before apheresis. The donor/recipient body weight ratio predicted the outcome of collection in a single large volume leukapheresis. Donors with body weight ratio >or=0.75 had 4.69 times higher likelihood to reach the minimum target cell dose. PMID- 19120368 TI - Prevalence of the JAK2 V617F mutation in patients with unprovoked venous thromboembolism of common sites and without overt myeloproliferative neoplasms. PMID- 19120369 TI - BCL2 protein expression in follicular lymphomas with t(14;18) chromosomal translocations. AB - The t(14;18)(q32;q21) chromosomal translocation induces BCL2 protein overexpression in most follicular lymphomas. However the expression of BCL2 is not always homogeneous and may demonstrate a variable degree of heterogeneity. This study analysed BCL2 protein expression pattern in 33 cases of t(14;18) positive follicular lymphomas using antibodies against two different epitopes (i.e. the widely used antibody BCL2/124 and an alternative antibody E17). 16/33 (49%) cases demonstrated strong BCL2 expression. In 10/33 (30%) cases, BCL2 expression was heterogeneous and in some of these, its loss appeared to be correlated with cell proliferation, as indicated by Ki67 expression. Double immunofluorescence labelling confirmed an inverse BCL2/Ki67 relationship, where in 24/28 (86%) cases cellular expression of BCL2 and Ki67 was mutually exclusive. In addition, seven BCL2 'pseudo-negative' cases were identified in which immunostaining was negative with antibody BCL2/124, but positive with antibody E17. Genomic DNA sequencing of these 'pseudo-negative' cases demonstrated eleven mutations in four cases and nine of these were missense mutations. It can be concluded that in follicular lymphomas, despite carrying the t(14;18) translocations, BCL2 protein expression may be heterogeneous and loss of BCL2 could be related to cell proliferation. Secondly, mutations in translocated BCL2 genes appear to be common and may cause BCL2 pseudo-negative immunostaining. PMID- 19120370 TI - Is refractory anaemia with ring sideroblasts and thrombocytosis (RARS-T) a necessary or useful diagnostic category? AB - Both the 2001 World Health Organisation (WHO) classification of haematopoietic neoplasms and the 2008 WHO classification revision include a distinctive diagnostic category, refractory anaemia with ring sideroblasts and thrombocytosis (RARS-T), to describe those rare patients who have both >or=15% ring sideroblasts and a sustained elevated platelet count. Recently, it has become clear that patients meeting WHO criteria for RARS-T have clonal JAK2(V617F) and MPL(W515) mutations at a similar rate to essential thrombocythaemia (ET). Given that the provisional classification of RARS-T as a myelodysplastic syndrome/myeloproliferative neoplasm (MDS/MPN) overlap syndrome, rather than as a form of MPN (i.e., ET), rests principally upon the presence of ring sideroblasts, which are a non-specific morphological finding, these new molecular results prompt reconsideration of the necessity for a distinctive RARS-T category. Here we review the historical developments that led up the definition of RARS-T as a disease entity, and we discuss conceptual understanding of RARS-T and arguments against continued use of RARS-T as a separate diagnostic category. PMID- 19120371 TI - Newer antifungal agents for invasive fungal infections in patients with haematological malignancy. AB - Since 2001 five new systemically administered antifungal agents have been approved for clinical use. This represents a major advance for antifungal therapy in haematological malignancy patients undergoing chemotherapy or haematopoietic stem cell transplant (HSCT). The echinocandins are a new class of antifungals with a novel mode of action. Capsofungin has already established itself as a valuable therapy for candidaemia and salvage therapy of invasive aspergillosis. Although both anidulafungin and micafungin are approved for treatment of candidiasis, their role in invasive aspergillosis requires more clinical trial evaluation. Of the two newer triazoles, voriconazole has been recommended in international guidelines as primary therapy for acute invasive aspergillosis. Posaconazole has a broad spectrum of activity in vitro and a potentially key role in antifungal prophylaxis in high-risk HSCT recipients and during prolonged neutropenia. Although some of these drugs have important interactions with other medications, and potential toxicities, they are safer to use and more efficacious than amphotericin B deoxycholate. Their arrival gives more choices to treat rarer mycoses and will facilitate clinical trial assessment of combination therapy of aspergillosis where single agent therapy gives less than 50% success rates. PMID- 19120372 TI - Dysregulation of innate immunity: hereditary periodic fever syndromes. AB - The hereditary periodic fever syndromes encompass a rare group of diseases that have lifelong recurrent episodes of inflammatory symptoms and an acute phase response in common. Clinical presentation can mimic that of lymphoproliferative disorders and patients often go undiagnosed for many years. These syndromes follow an autosomal inheritance pattern, and the major syndromes are linked to specific genes, most of which are involved in regulation of the innate immune response through pathways of apoptosis, nuclear factor kappaBeta activation and cytokine production. In others, the link between the protein involved and inflammation is less clear. The recurrent inflammation can lead to complications, such as renal impairment due to amyloidosis and vasculitis, visual impairment, hearing loss, and joint destruction, depending on the specific syndrome. In recent years, treatment options for these diseases have improved significantly. Early establishment of an accurate diagnosis and start of appropriate therapy improves prognosis in these patients. PMID- 19120373 TI - Ependymoblastoma: dear, damned, distracting diagnosis, farewell!*. AB - Ependymoblastoma is a diagnostic label that has been applied to a variety of rare central nervous system (CNS) tumors over the last eight decades. Consequently, there is uncertainty about whether such an entity exists and what its characteristic features might be. The current study, based on 14 cases from our institutional archives and identified by the search terms "ependymoblastoma,"ependymoblastomatous," "ependymoblastic" or "PNET with ependymal differentiation," aimed to test the hypothesis that the ependymoblastoma is a distinct and recognizable entity. Ependymoblastic rosettes are a key diagnostic feature and were present in 11/14 (79%) tumors, eight (73%) of which were embryonal tumors with abundant areas of neuropil-like differentiation. Three other cases showed rare ependymoblastic rosettes in the histopathological setting of a typical primitive neuroectodermal tumor (PNET), medulloblastoma (MB) or atypical teratoid/rhabdoid tumor (AT/RT). The remaining cases were all embryonal tumors with structures that mimicked ependymoblastic rosettes. Our results indicate that ependymoblastic rosettes are most frequently encountered in embryonal tumors with abundant neuropil and less frequently in other CNS embryonal neoplasms, including PNET, MB and AT/RT. We believe that ependymoblastoma as a diagnosis is neither precise nor specific and that it is time once and for all to retire this diagnosis from the lexicon of neuropathology. PMID- 19120374 TI - Health-related quality of life in women with breast cancer in Belgrade (Serbia). PMID- 19120375 TI - Breast MRI in a case of "early onset" lactating adenoma. PMID- 19120376 TI - Breast cancer and multiple myeloma at initial presentation. PMID- 19120377 TI - Clinical presentation of primary actinomycosis of the breast. PMID- 19120378 TI - Resource use and cost of diagnostic workup of women with suspected breast cancer. AB - We estimated resource use and costs associated with a diagnostic workup for suspected breast cancer among Medicare beneficiaries. Using Medicare claims data, we found that the average cost of a diagnostic workup for suspected breast cancer -whether it eventuated in a breast cancer diagnosis or not--was $361, and did not vary by presentation (signs/symptoms or screening mammography). In the aggregate, we estimate that Medicare spends approximately $679 million annually on diagnostic workups for women with suspected breast cancer, and that false positive mammograms result in diagnostic costs of approximately $250 million. PMID- 19120379 TI - Breast cancer: unique communication challenges and strategies to address them. AB - Women with breast cancer have become increasingly more involved on a national and local level in advocating for resources to fight cancer. However, in the context of the relationship with their physicians and other cancer caregivers, much remains to be done in providing them with adequate support. In this paper, we highlight the difficulties in communication related to breast cancer and describe strategies and approaches that may be helpful in improving the communication throughout the cancer trajectory. Specifically, breast cancer patients have high unmet information needs relevant to health information and dissatisfaction with the actual information they receive from their providers. These needs seem even more pronounced when patients are older, of lower socio-economic class and from differing cultural backgrounds which may affect their ability to express their desires for information and desire to be involved in decision-making about their treatment. Other communication challenges can be envisioned as occurring at key points across the cancer trajectory: diagnosis disclosure, treatment failure, transition to palliative care, and end of life discussions. These involve techniques as basic as how to establish trust and rapport and determine a patient's information and decision-making preferences and as complex as giving bad news. These strategies are now viewed as essential skills in that they can affect patient distress and quality of life, satisfaction, and malpractice litigation as well as practitioner stress and burnout. PMID- 19120380 TI - Psychosocial consequences of genetic counseling: a population-based follow-up study. AB - We aimed to examine the psychosocial impact of genetic counseling for hereditary breast and ovarian cancer 1 year following genetic counseling. We conducted a population-based prospective follow-up study of 213 women who received genetic counseling for hereditary breast and ovarian cancer, 319 women who underwent mammography (Reference Group I), and a random sample of 1,070 women from the general population (Reference Group II). The prevalence of anxiety decreased by 4.7% (95% CI: -3.5; 12.8) from baseline to 1 year of follow-up in the Genetic Counseling Group. Likewise, it decreased by 2.5% (95% CI: -4.5; 9.5) in Reference Group I and by 1.1% (95% CI: -2.3; 4.7) in Reference Group II. The prevalence of depression increased equally (1-3%) in the three study groups. 52% of the women referred for genetic counseling experienced cancer-specific distress at baseline and this proportion decreased to 41% after 12 months of follow-up. This decrease of 10.8% (95% CI: 1.4; 20.8) exceeded the decrease observed in both Reference Groups. However, it was statistically significant only in the case of Reference Group II (p=0.006). Our findings indicate that genetic counseling can help alleviate cancer-specific distress among women with a family history of breast and ovarian cancer. Further, genetic counseling does not appear to have an adverse impact on general anxiety, symptoms of depression, or health-related quality of life. PMID- 19120381 TI - Effects of exercise on quality of life in women living with breast cancer: a systematic review. AB - The goal of this systematic review was to examine the effect of exercise on quality of life (QOL) in women living with breast cancer. Data bases searched were MEDLINE, EMBASE, CINAHL, PubMed, and PEDro. Keywords were "breast cancer and quality of life" in combination with "exercise" or "physical activity" (with associated Mesh terms). Limits were English or French language. Included studies were independently reviewed for methodological quality (van Tulder et al.) and assigned a level of evidence (Centre for Evidence-Based Medicine). Nine relevant randomized controlled trials were included: four of moderate methodological quality and five of high methodological quality. There was strong evidence that exercise positively influences QOL in women living with breast cancer. Exercise can be an effective strategy to improve QOL in women living with breast cancer. Future research is necessary to determine optimal exercise types and parameters. PMID- 19120382 TI - The structural landscape of the health care system for breast cancer care: results from the Los Angeles Women's Health Study. AB - The structure of health care has been rapidly evolving in response to financial pressures and demands to improve quality. Little work has documented the structure of care and its impact in the context of breast cancer care. We conducted a survey to characterize Los Angeles physicians caring for breast cancer patients and the structural landscape of the healthcare system in which they practice. Cross-sectional survey of physicians who treated a population based cohort of breast cancer patients. We surveyed 477 physicians, targeting all Los Angeles County medical oncologists, radiation oncologists, and surgeons reported by patients participating in the Los Angeles Women's Health Study (77% response rate). Specialty-specific questionnaires were developed. Items were based on the structure and quality of care literature, cognitive interviews with cancer care specialists, and existing physician survey instruments. Breast cancer care providers in Los Angeles are diverse, with one-third non-white and 46% speaking a non-English language. Group practice is most common, (37% single specialty, 16% group-model HMO, 8% multi-specialty group). Minimal teaching involvement predominates. Mean new breast cancer patient volumes are relatively high (8 per month overall; six for surgeons), representing 46% of new cancer patients. Physicians reported high career satisfaction levels (83-92%). Physicians were least satisfied with the amount of time spent with patients (82%). Data from this study represent important building blocks for further analyses to determine the impact of structural characteristics on the quality of care that breast cancer patient's experience. PMID- 19120383 TI - Breast-conservation treatment outcomes: a community hospital's experience. AB - In the United States, the majority of early breast cancer patients choose breast conserving treatment in the community setting, yet there is a paucity of literature describing outcomes. In this paper, we describe our experience with breast-conserving treatment in a small community hospital. Our hospital tumor registry was used to identify breast cancer cases diagnosed at our hospital between 1997 and 2003. We limited our study to those women with initial attempts at breast-conserving surgery (BCS) who had follow-up oncology treatment at on campus affiliated oncological services. We looked at factors that influence survival for early stage 0-II disease such as tumor and patient characteristics, completeness of local surgical tumor excision, and adjuvant treatment. We also evaluated the percentage of cases in which the initial BCS did not achieve adequate surgical oncological results and the number and type of subsequent surgeries that were required to achieve this goal. There were 185 cases with a median patient age of 55 and a median follow-up time of 53 months. Most tumors were stage 0-I (68%) or stage II (23%). A single surgery was deemed sufficient to achieve the desired oncological outcome in 54% of cases; the remaining cases (46%) required additional surgeries. A final margin of 5 mm or greater was successfully achieved in 81% of cases. Ninety-two percent of the patients underwent radiotherapy, 65% received hormonal therapy, and 49% underwent chemotherapy. One hundred and sixty one patients had successful breast-conserving surgeries (87%) and 24 patients (13%) ultimately required mastectomy. There were four loco-regional recurrences and 19 deaths during the study period. Our disease free survival rate for early-stage cancer (stage 0-II) was 91% at 5 years. Our study shows that high-quality patient outcomes for breast-conserving treatment can be achieved in the community setting. PMID- 19120384 TI - Lupus vulgaris due to Mycobacterium bovis bacillus Calmette-Guerin (BCG) at the site of previous BCG vaccination. PMID- 19120385 TI - Omphalith. PMID- 19120386 TI - Herpes zoster complicated by delayed intracranial haemorrhage. PMID- 19120387 TI - Narrowband ultraviolet B phototherapy for inflammatory vitiligo with raised borders associated with Sjogren's syndrome. PMID- 19120388 TI - Cutaneous giant plasmacytoma associated with monoclonal gammopathy of unknown significance. PMID- 19120389 TI - Plantar congenital fibrolipomatous hamartomas. PMID- 19120390 TI - Long-term control of recalcitrant psoriasis with combination infliximab and methotrexate. PMID- 19120391 TI - Recall injection-site reactions to etanercept in a patient with psoriasis. PMID- 19120392 TI - DRESS syndrome caused by efalizumab: comment. PMID- 19120393 TI - Targetoid pattern of hair regrowth in alopecia areata: a case report. PMID- 19120394 TI - Contact allergy to ketoconazole cross-sensitive to miconazole. PMID- 19120395 TI - Chromosome 22q11.2 deletion syndrome associated with severe eczema. PMID- 19120396 TI - Epidermotropic skin metastasis from gastric cancer: immunohistochemical analysis using cytokeratins. PMID- 19120397 TI - Trichomegaly of the eyelashes following treatment with cetuximab. PMID- 19120398 TI - Generalized livedo reticularis as the first sign of metastatic breast carcinoma. PMID- 19120399 TI - Subungueal pyogenic granuloma secondary to docetaxel therapy. PMID- 19120400 TI - Nephrogenic systemic fibrosis in a Chinese renal-transplant recipient. PMID- 19120401 TI - A case of solitary collagenoma localized on the upper lip mimicking mucocele. PMID- 19120402 TI - Melanonychia caused by Stenotrophomonas maltophilia. PMID- 19120403 TI - Inflammatory nodules around the axilla: an uncommon localization of orf virus infection. PMID- 19120406 TI - Development of executive function: more than conscious reflection. PMID- 19120405 TI - A hierarchical competing systems model of the emergence and early development of executive function. AB - The hierarchical competing systems model (HCSM) provides a framework for understanding the emergence and early development of executive function--the cognitive processes underlying the conscious control of behavior--in the context of search for hidden objects. According to this model, behavior is determined by the joint influence of a developmentally invariant habit system and a conscious representational system that becomes increasingly influential as children develop. This article describes a computational formalization of the HCSM, reviews behavioral and computational research consistent with the model, and suggests directions for future research on the development of executive function. PMID- 19120407 TI - When in competition against engrained habits, is conscious representation sufficient or is inhibition of the habit also needed? PMID- 19120408 TI - Dynamic executives. PMID- 19120410 TI - Cue salience and infant perseverative reaching: tests of the dynamic field theory. AB - Skilled behavior requires a balance between previously successful behaviors and new behaviors appropriate to the present context. We describe a dynamic field model for understanding this balance in infant perseverative reaching. The model predictions are tested with regard to the interaction of two aspects of the typical perseverative reaching task: the visual cue indicating the target and the memory demand created by the delay imposed between cueing and reaching. The memory demand was manipulated by imposing either a 0- or a 3-second delay, and the salience of the cue to reach was systematically varied. Infants demonstrated fewer perseverative errors at 0-delay versus 3-second delay based on the cue salience, such that a more salient visual cue was necessary to overcome a longer delay. These results have important implications for understanding both the basic perceptual-motor processes that produce reaching in infants and skilled flexible behavior in general. PMID- 19120411 TI - Identifying unreliable informants: do children excuse past inaccuracy? AB - In three experiments (N = 123; 148; 28), children observed a video in which two speakers offered alternative labels for unfamiliar objects. In Experiment 1, 3- to 5-year-olds endorsed the label given by a speaker who had previously labeled familiar objects accurately, rather than that given by a speaker with a history of inaccurate labeling, even when the inaccurate speaker erred only while blindfolded. In Experiments 2 and 3, 3- to 7-year-olds showed no preference for the label given by a previously inaccurate but blindfolded speaker, over that given by a second inaccurate speaker with no obvious excuse for erring. Children based their endorsements on speakers' history of accuracy or inaccuracy irrespective of the speakers' information access at the time, raising doubts that children made mentalistic interpretations of speakers' inaccuracy. PMID- 19120412 TI - Early bilingualism enhances mechanisms of false-belief reasoning. AB - In their first years, children's understanding of mental states seems to improve dramatically, but the mechanisms underlying these changes are still unclear. Such 'theory of mind' (ToM) abilities may arise during development, or have an innate basis, developmental changes reflecting limitations of other abilities involved in ToM tasks (e.g. inhibition). Special circumstances such as early bilingualism may enhance ToM development or other capacities required by ToM tasks. Here we compare 3-year-old bilinguals and monolinguals on a standard ToM task, a modified ToM task and a control task involving physical reasoning. The modified ToM task mimicked a language-switch situation that bilinguals often encounter and that could influence their ToM abilities. If such experience contributes to an early consolidation of ToM in bilinguals, they should be selectively enhanced in the modified task. In contrast, if bilinguals have an advantage due to better executive inhibitory abilities involved in ToM tasks, they should outperform monolinguals on both ToM tasks, inhibitory demands being similar. Bilingual children showed an advantage on the two ToM tasks but not on the control task. The precocious success of bilinguals may be associated with their well-developed control functions formed during monitoring and selecting languages. PMID- 19120413 TI - Neurocognitive development of relational reasoning. AB - Relational reasoning is an essential component of fluid intelligence, and is known to have a protracted developmental trajectory. To date, little is known about the neural changes that underlie improvements in reasoning ability over development. In this event-related functional magnetic resonance imaging (fMRI) study, children aged 8-12 and adults aged 18-25 performed a relational reasoning task adapted from Raven's Progressive Matrices. The task included three levels of relational reasoning demands: REL-0, REL-1, and REL-2. Children exhibited disproportionately lower accuracy than adults on trials that required integration of two relations (REL-2). Like adults, children engaged lateral prefrontal cortex (PFC) and parietal cortex during task performance; however, they exhibited different time courses and activation profiles, providing insight into their approach to the problems. As in prior studies, adults exhibited increased rostrolateral PFC (RLPFC) activation when relational integration was required (REL-2 > REL-1, REL-0). Children also engaged RLPFC most strongly for REL-2 problems at early stages of processing, but this differential activation relative to REL-1 trials was not sustained throughout the trial. These results suggest that the children recruited RLPFC while processing relations, but failed to use it to integrate across two relations. Relational integration is critical for solving a variety of problems, and for appreciating analogies; the current findings suggest that developmental improvements in this function rely on changes in the profile of engagement of RLPFC, as well as dorsolateral PFC and parietal cortex. PMID- 19120415 TI - Co-speech gesture as input in verb learning. AB - People gesture a great deal when speaking, and research has shown that listeners can interpret the information contained in gesture. The current research examines whether learners can also use co-speech gesture to inform language learning. Specifically, we examine whether listeners can use information contained in an iconic gesture to assign meaning to a novel verb form. Two experiments demonstrate that adults and 2-, 3-, and 4-year-old children can infer the meaning of novel intransitive verbs from gestures when no other source of information is present. The findings support the idea that gesture might be a source of input available to language learners. PMID- 19120414 TI - Developmental changes in visual object recognition between 18 and 24 months of age. AB - Two experiments examined developmental changes in children's visual recognition of common objects during the period of 18 to 24 months. Experiment 1 examined children's ability to recognize common category instances that presented three different kinds of information: (1) richly detailed and prototypical instances that presented both local and global shape information, color, textural and featural information, (2) the same rich and prototypical shapes but no color, texture or surface featural information, or (3) that presented only abstract and global representations of object shape in terms of geometric volumes. Significant developmental differences were observed only for the abstract shape representations in terms of geometric volumes, the kind of shape representation that has been hypothesized to underlie mature object recognition. Further, these differences were strongly linked in individual children to the number of object names in their productive vocabulary. Experiment 2 replicated these results and showed further that the less advanced children's object recognition was based on the piecemeal use of individual features and parts, rather than overall shape. The results provide further evidence for significant and rapid developmental changes in object recognition during the same period children first learn object names. The implications of the results for theories of visual object recognition, the relation of object recognition to category learning, and underlying developmental processes are discussed. PMID- 19120416 TI - To generalize or not to generalize: spatial categories are influenced by physical attributes and language. AB - The current work explored the conditions under which infants generalize spatial relationships from one event to another. English-learning 5-month-olds habituated to a tight- or loose-fit covering event dishabituated to a change in fit during a containment test event, but infants habituated to a visually similar occlusion event did not. Thus, infants' responses appeared to be driven by the physical nature of the fit rather than visual similarity. This response pattern was replicated with Korean-speaking adults, but English-speaking adults showed no sensitivity to change in fit for either event. These findings suggest that language development links linguistic forms to universal, pre-existing representations of meaning, and that linguistic experience can shape sensitivity to distinctions that are marked in one's native language. PMID- 19120417 TI - Toddlers can adaptively change how they categorize: same objects, same session, two different categorical distinctions. AB - Two experiments demonstrate that 14- to 18-month-old toddlers can adaptively change how they categorize a set of objects within a single session, and that this ability is related to vocabulary size. In both experiments, toddlers were presented with a sequential touching task with objects that could be categorized either according to some perceptually salient dimension corresponding to a taxonomic distinction (e.g. animals vs. vehicles) or to some less obvious dimension (e.g. rigid vs. deformable). In each experiment, children with larger productive vocabularies responded to both dimensions, showing evidence of sensitivity to each way of categorizing the items. Children with smaller productive vocabularies attended only to the taxonomically related categorical grouping. These experiments confirm that toddlers can adaptively shift the basis of their categorization and highlight the dynamic interaction between the child and the current task in early categorization. PMID- 19120418 TI - Training and transfer effects of executive functions in preschool children. AB - Executive functions, including working memory and inhibition, are of central importance to much of human behavior. Interventions intended to improve executive functions might therefore serve an important purpose. Previous studies show that working memory can be improved by training, but it is unknown if this also holds for inhibition, and whether it is possible to train executive functions in preschoolers. In the present study, preschool children received computerized training of either visuo-spatial working memory or inhibition for 5 weeks. An active control group played commercially available computer games, and a passive control group took part in only pre- and posttesting. Children trained on working memory improved significantly on trained tasks; they showed training effects on non-trained tests of spatial and verbal working memory, as well as transfer effects to attention. Children trained on inhibition showed a significant improvement over time on two out of three trained task paradigms, but no significant improvements relative to the control groups on tasks measuring working memory or attention. In neither of the two interventions were there effects on non-trained inhibitory tasks. The results suggest that working memory training can have significant effects also among preschool children. The finding that inhibition could not be improved by either one of the two training programs might be due to the particular training program used in the present study or possibly indicate that executive functions differ in how easily they can be improved by training, which in turn might relate to differences in their underlying psychological and neural processes. PMID- 19120419 TI - What makes relational reasoning smart? Revisiting the perceptual-to-relational shift in the development of generalization. AB - Development of reasoning is often depicted as involving increasing use of relational similarities and decreasing use of perceptual similarities ('the perceptual-to-relational shift'). We argue that this shift is a special case of a broader developmental trend: increasing sensitivity to the predictive accuracy of different similarity types. To test this hypothesis, we asked participants (3-, 4 , 5-year-olds and adults) to generalize novel information on two types of problems--offspring problems, where relational matches yield accurate generalizations, and prey problems, where perceptual matches yield accurate generalizations. On offspring problems, we replicated prior findings of increasing relational matches with age. However, we observed decreasing relational matches on prey problems. Provided feedback on their responses, 3-year olds showed the same trend. Findings suggest that the relational shift commonly observed in categorization and analogical reasoning may reflect a general increase in children's sensitivity to cue validity rather than an overall preference to generalize over perceptual similarity. PMID- 19120420 TI - Logic and belief across the lifespan: the rise and fall of belief inhibition during syllogistic reasoning. AB - Popular reasoning theories postulate that the ability to inhibit inappropriate beliefs lies at the heart of the human reasoning engine. Given that people's inhibitory capacities are known to rise and fall across the lifespan, we predicted that people's deductive reasoning performance would show similar curvilinear age trends. A group of children (12-year-olds), young adults (20-year olds), and older adults (65+-year-olds) were presented with a classic syllogistic reasoning task and a decision-making questionnaire. Results indicated that on syllogisms where beliefs and logic conflicted, reasoning performance showed the expected curvilinear age trend: Reasoning performance initially increased from childhood to early adulthood but declined again in later life. On syllogisms where beliefs and logic were consistent and sound reasoning did not require belief inhibition, however, age did not affect performance. Furthermore, across the lifespan we observed that the better people were at resisting intuitive temptations in the decision-making task, the less they were biased by their beliefs on the conflict syllogisms. As with the effect of age, one's ability to override intuitions in the decision-making task did not mediate reasoning performance on the no-conflict syllogisms. Results lend credence to the postulated central role of inhibitory processing in those situations where beliefs and logic conflict. PMID- 19120421 TI - At 11 months, prosody still outranks statistics. AB - English-learning 7.5-month-olds are heavily biased to perceive stressed syllables as word onsets. By 11 months, however, infants begin segmenting non-initially stressed words from speech. Using the same artificial language methodology as Johnson and Jusczyk (2001), we explored the possibility that the emergence of this ability is linked to a decreased reliance on prosodic cues to word boundaries accompanied by an increased reliance on syllable distribution cues. In a baseline study, where only statistical cues to word boundaries were present, infants exhibited a familiarity preference for statistical words. When conflicting stress cues were added to the speech stream, infants exhibited a familiarity preference for stress as opposed to statistical words. This was interpreted as evidence that 11-month-olds weight stress cues to word boundaries more heavily than statistical cues. Experiment 2 further investigated these results with a language containing convergent cues to word boundaries. The results of Experiment 2 were not conclusive. A third experiment using new stimuli and a different experimental design supported the conclusion that 11-month-olds rely more heavily on prosodic than statistical cues to word boundaries. We conclude that the emergence of the ability to segment non-initially stressed words from speech is not likely to be tied to an increased reliance on syllable distribution cues relative to stress cues, but instead may emerge due to an increased reliance on and integration of a broad array of segmentation cues. PMID- 19120422 TI - Recognizing the role of perception in action at 6 months. AB - The present research examined whether infants as young as 6 months of age would consider what objects a human agent could perceive when interpreting her actions on the objects. In two experiments, the infants took the agent's actions of repeatedly reaching for and grasping one of two possible objects as suggesting her preference for that object only when the agent could detect both objects, not when the agent's perceptual access to the second object was absent, either because a large screen hid the object from the agent (Experiment 1), or because the agent sat with her back toward the object (Experiment 2). These results suggest that young infants recognize the role of perception in constraining an agent's goal-actions. PMID- 19120423 TI - The relationship between second-order false belief and display rules reasoning: the integration of cognitive and affective social understanding. AB - To investigate the relation between cognitive and affective social understanding, Japanese 4- to 8-year-olds received tasks of first- and second-order false beliefs and prosocial and self-presentational display rules. From 6 to 8 years, children comprehended display rules, as well as second-order false belief, using social pressures justifications decreasingly and motivational justifications with embedded perspectives increasingly with age. Although not related to either type of display across ages, second-order tasks were associated with both types of display tasks only at 8 years when examined in each age group. Results suggest that children base their second-order theory of mind and display rules understanding on distinct reasoning until middle childhood, during which time the originally distinct aspects of social understanding are integrated. PMID- 19120424 TI - Behavioral genetic analyses of prosocial behavior in adolescents. AB - Prosocial behavior is an important aspect of normal social and psychological development. Adult and child twin studies typically estimate the heritability of prosocial behavior to be between 30 and 50%, although relatively little is known about genetic and environmental influences upon prosocial behavior in adolescence. We therefore examined reports of prosocial behavior in a large longitudinal family study of 1160 adolescent twin pairs (aged between 13 and 19 years). Prosocial behavior was assessed at two time points by self-report and at the second time point by additional parent-ratings using the Strengths and Difficulties Questionnaire (SDQ; Goodman, 1997). Adolescent females were reported to be significantly more prosocial than males (p < .001). Univariate analyses primarily showed moderate heritability and large nonshared environmental influences. There was a moderate genetic correlation between self- and parent reported prosocial behaviour, suggesting that both types of rater were tapping into genetically overlapping constructs. Longitudinal analyses revealed that continuity was largely explained by genes. Unique environmental influences were predominantly time-specific and were the major source of individual differences. PMID- 19120425 TI - Motor and cognitive control in a normative sample of 7-year-olds. AB - The aim of the present study was to explore the relationship between cognitive and motor control by correlating individual performance on a variety of complex tasks in a normative sample of over 100 7-year-olds. While there are a few studies including children with specific developmental disorders (i.e. ADHD and DCD) showing that they share similar comorbid deficits, it is still unclear whether control processes share variance in normative samples. Results presented here reveal on the one hand that some aspects of cognitive and motor control are significantly interrelated, even when controlling for age. On the other hand, individuals' ability to efficiently respond to the speed and accuracy demands of any of the included tasks appeared to constitute an important aspect of executive functioning. PMID- 19120426 TI - Early gesture selectively predicts later language learning. AB - The gestures children produce predict the early stages of spoken language development. Here we ask whether gesture is a global predictor of language learning, or whether particular gestures predict particular language outcomes. We observed 52 children interacting with their caregivers at home, and found that gesture use at 18 months selectively predicted lexical versus syntactic skills at 42 months, even with early child speech controlled. Specifically, number of different meanings conveyed in gesture at 18 months predicted vocabulary at 42 months, but number of gesture+speech combinations did not. In contrast, number of gesture+speech combinations, particularly those conveying sentence-like ideas, produced at 18 months predicted sentence complexity at 42 months, but meanings conveyed in gesture did not. We can thus predict particular milestones in vocabulary and sentence complexity at age by watching how children move their hands two years earlier. PMID- 19120427 TI - Preschoolers continue to trust a more accurate informant 1 week after exposure to accuracy information. AB - To determine whether children retain a preference for a previously accurate informant only in the short term or for long-term use, 3- and 4-year-old children were tested in two experiments. In both experiments, children were given accuracy information about two informants and were subsequently tested for their selective trust in the two informants (Experiment 1: immediately, 1 day and 1 week later; Experiment 2: immediately, 4 days and 1 week later). Both age groups preferred to trust the accurate informant not only immediately after receiving accuracy information but also at subsequent time-points. Children who were immediately able to explicitly identify the accurate informant were significantly more likely to seek and accept information from her 1 week later. However, even when they had not been asked to explicitly identify the accurate informant both age groups still maintained their preference for her. Thus, by 3 years of age, children spontaneously choose a previously accurate informant up to 1 week after exposure to information regarding her accuracy. PMID- 19120428 TI - Biological and environmental initial conditions shape the trajectories of cognitive and social-emotional development across the first years of life. AB - Human development is thought to evolve from the dynamic interchange of biological dispositions and environmental provisions; yet the effects of specific biological and environmental birth conditions on the trajectories of cognitive and social emotional growth have rarely been studied. We observed 126 children at six time points from birth to 5 years. Intelligence, maternal sensitivity, and child social engagement were repeatedly tested. Effects of neonatal vagal tone (VT) and maternal postpartum depressive symptoms on growth-rates were assessed. Cognitive development showed a substantial growth-spurt between 2 and 5 years and social engagement increased rapidly across the first year and more gradually thereafter. VT improved cognitive and social-emotional growth-rates across the first year, whereas maternal depressive symptoms interfered with growth from 2 to 5 years. Differences between infants with none, one, or two non-optimal birth conditions increased with age. Findings shed light on the dynamics of early development as it is shaped by biological and environmental initial conditions. PMID- 19120429 TI - The narrow fellow in the grass: human infants associate snakes and fear. AB - Why are snakes such a common target of fear? One current view is that snake fear is one of several innate fears that emerge spontaneously. Another is that humans have an evolved predisposition to learn to fear snakes. In the first study reported here, 9- to 10-month-old infants showed no differential spontaneous reaction to films of snakes versus other animals. In the second study, 7- to 18 month-old infants associated snakes with fear: As predicted, they looked longer at films of snakes while listening to a frightened human voice than while listening to a happy voice. In the third study, infants did not look differentially to still photos of snakes and other animals, indicating that movement is crucial to infants' association of snakes with fear. These results offer support for the view that humans have a natural tendency to selectively associate snakes with fear. PMID- 19120430 TI - Insulin analogues: a step forward in the care in diabetes. PMID- 19120431 TI - Insulin analogues: an example of applied medical science. AB - Insulin analogues were developed to try and achieve more physiological insulin replacement from injection in the subcutaneous site. Their pharmacokinetics and pharmacodynamics differ from human insulin when injected subcutaneously because of alterations in the amino acid sequence of the insulin molecule. The rapid acting insulin analogues, lispro, aspart and glulisine, have a rapid onset of action and shorter duration of action because of changes to the B26-30 portion of insulin inhibiting formation of dimers and hexamers. They appear to improve postprandial glucose, incidence of hypoglycaemia and patient satisfaction and, when used in combination with basal insulin analogues, improve glycosylated haemoglobin in comparison to conventional insulin therapy. Additionally, they have been successfully used in children, pregnant women, in pump therapy and as part of premixed biphasic regimens. The two basal insulin analogues, glargine and detemir, developed by adjusting the isoelectric point and adding a fatty acid residue, respectively, have a protracted duration of action and a relatively smooth profile. Their pharmacokinetic and pharmacodynamic profiles have been assessed using euglycaemic clamp protocols. Both analogues have a longer duration of action, less of a peak of activity and a reduced variability with repeated injection. There is some evidence to suggest that detemir may have a slight hepatoselective effect. Clinical studies have shown a lower relative risk of hypoglycaemia and detemir appears to have a weight-sparing action. Insulin analogues represent a successful example of applied medical science. PMID- 19120432 TI - The rapid-acting properties of insulin aspart are preserved in elderly people with type 2 diabetes. AB - AIMS: Elderly type 2 diabetes patients may face increased risk of hypoglycaemic episodes because of unpredictable eating habits. The pharmacokinetics and pharmacodynamics of insulin aspart (IAsp) were studied in elderly patients to examine the potential for postprandial dosing. METHODS: Nineteen type 2 diabetes subjects, aged > or =65 years, were enrolled in this randomized, double-blind, crossover trial. Subjects received 0.3 U/kg IAsp or regular human insulin during a glucose clamp procedure. RESULTS: IAsp showed a faster onset of action with significantly higher values for area under the glucose infusion rate curves, AUC(GIR(0-120 min)) and AUC(GIR(0-300 min)) (p = 0.0001). Maximum metabolic activity was higher (4.4 vs. 3.8 mg/kg/min, p = 0.0039) and occurred earlier with IAsp (196 vs. 278 min, p < 0.0001). Late metabolic activity (AUC(GIR (300-600 min))) was significantly lower with IAsp (p = 0.0006). CONCLUSION: Clinical studies are required to confirm whether postprandial administration of IAsp is appropriate for elderly patients. PMID- 19120433 TI - Once-daily insulin detemir is comparable to once-daily insulin glargine in providing glycaemic control over 24 h in patients with type 2 diabetes: a double blind, randomized, crossover study. AB - Once-daily dosing with insulin detemir and insulin glargine were compared in a double-blind, randomised, crossover study in type 2 diabetes subjects previously treated with other antihyperglycaemic medications. Blood glucose was measured through continuous glucose monitoring (CGM). Insulin dose was adjusted daily during the titration phase to achieve target blood glucose values of (70-120 mg/dL) during the basal period, defined as 2400-0600 hours. The last meal of the day started at 1800 h and basal insulin was injected at 2000 h. The CGM data for a 24-h period on the second consecutive day after achieving target blood glucose levels were compared between treatments. Twenty-nine subjects completed the study. Over a 24-h measurement period, once-daily dosing with insulin detemir provided glycaemic control very similar to that of once-daily insulin glargine in patients with type 2 diabetes after both had been titrated to the same glucose target. Insulin detemir- and insulin glargine-treated subjects had similar mean 24-h glucose values (133 +/- 21 mg/dL compared with 126+/-20 mg/dL respectively, p = 0.385) and similar glucose values during the basal period (105 +/- 23 mg/dL compared with 98 +/- 19 mg/dL, respectively p = 0.204).Target basal glycaemic control was achieved in all subjects in a mean of 3.8 days for detemir and 3.5 days for glargine (p = 0.360). The mean dose of detemir was similar to that of glargine (26.3 and 26.6 units/day, respectively, p = 0.837). In this study, once daily dosing of insulin detemir provided 24-h glycaemic control similar to that of insulin glargine in patients with type 2 diabetes. PMID- 19120437 TI - Balancing excitation, inhibition and endocannabinoids (Commentary on Ademark et al.). PMID- 19120438 TI - Endocannabinoid-dependent plasticity at GABAergic and glutamatergic synapses in the striatum is regulated by synaptic activity. AB - Long-term depression (LTD) at striatal synapses is mediated by postsynaptic endocannabinoid (eCB) release and presynaptic cannabinoid 1 receptor (CB(1)R) activation. Previous studies have indicated that eCB mobilization at excitatory synapses might be regulated by afferent activation. To further address the role of neuronal activity in synaptic plasticity we examined changes in synaptic strength induced by the L-type calcium channel activator 2,5-dimethyl-4-[2 (phenylmethyl)benzoyl]-1H-pyrrole-3-carboxylic acid methyl ester (FPL 64176, FPL) at glutamatergic and gamma-aminobutyric acid (GABA)ergic synapses in the striatum. We found that the basic mechanisms for FPL-mediated eCB signaling are the same at glutamatergic and GABAergic synapses. FPL-induced LTD (FPL-LTD) was blocked in slices treated with the CB(1)R antagonist AM251 (2 microm), but established depression was not reversed by AM251. FPL-LTD was temperature dependent, blocked by protein translation inhibitors and prevented by intracellular loading of the anandamide transporter inhibitor VDM11 (10 microm) at both glutamatergic and GABAergic synapses. FPL-LTD at glutamatergic synapses required paired-pulse afferent stimulation, while FPL-LTD at GABAergic synapses could be induced even in the absence of explicit afferent activation. By evaluating tetrodotoxin-insensitive spontaneous inhibitory postsynaptic currents we found that neuronal firing is vital for eCB release and LTD induction at GABAergic synapses, but not for short-term depression induced by CB(1)R agonist. The data presented here suggest that the level of neuronal firing regulates eCB signaling by modulating release from the postsynaptic cell, as well as interacting with presynaptic mechanisms to induce LTD at both glutamatergic and GABAergic synapses in the striatum. PMID- 19120440 TI - Involvement of mitochondrial signaling pathways in the mechanism of Fas-mediated apoptosis after spinal cord injury. AB - Activation of the Fas receptor has been recently linked to apoptotic cell death after spinal cord injury (SCI). Although it is generally considered that Fas activation mediates apoptosis predominantly through the extrinsic pathway, we hypothesized that intrinsic mitochondrial signaling could be involved in the underlying mechanism of Fas-induced apoptosis after SCI. In the present study, we utilized the Fejota clip compression model of SCI at T5-6 in C57BL/6 Fas deficient (lpr) and wild-type mice. Complementary studies were conducted using an in vitro model of trauma or a Fas-activating antibody to induce apoptosis in primary neuronal-glial mixed spinal cord cultures. After in vivo SCI, lpr mice, in comparison with wild-type mice, exhibited reduced numbers of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells at the lesion, reduced expression of truncation of Bid (tBid), apoptosis-inducing factor, activated caspase-9 and activated caspase-3, and increased expression of the antiapoptotic proteins Bcl-2 and Bcl-xL. After in vitro neurotrauma or the induction of Fas signaling by the Jo2 activating antibody, lpr spinal cord cultures showed an increased proportion of cells retaining mitochondrial membrane integrity and a reduction of tBid expression, caspase-9 and caspase-3 activation, and TUNEL-positive cells as compared to wild-type spinal cord cultures. The neutralization of Fas ligand (FasL) protected against traumatically induced or Fas-mediated caspase-3 activation and the loss of mitochondrial membrane potential and tBid expression in wild-type spinal cord cultures. However, in lpr spinal cord cultures, FasL neutralization had no protective effects. In summary, these data provide direct evidence for the induction of intrinsic mitochondrial signaling pathways following Fas activation after SCI. PMID- 19120439 TI - Calcyon is necessary for activity-dependent AMPA receptor internalization and LTD in CA1 neurons of hippocampus. AB - Calcyon is a single transmembrane endocytic protein that regulates clathrin assembly and clathrin-mediated endocytosis in the brain. Ultrastructural studies indicate that calcyon localizes to spines, but whether it regulates glutamate neurotransmission is not known. Here, we show that deletion of the calcyon gene in mice inhibits agonist-stimulated endocytosis of alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid receptors (AMPARs), without altering basal surface levels of the GluR1 or GluR2 subunits. Whole-cell patch-clamp studies of hippocampal neurons in culture and CA1 synapses in slices revealed that knockout (KO) of calcyon abolishes long-term synaptic depression (LTD), whereas mini analysis in slices indicated basal transmission in the hippocampus is unaffected by the deletion. Further, transfection of green fluorescent protein-tagged calcyon rescued the ability of KO cultures to undergo LTD. In contrast, intracellular dialysis of a fusion protein containing the clathrin light-chain binding domain of calcyon blocked the induction of LTD in wild-type hippocampal slices. Taken together, the present studies involving biochemical, immunological and electrophysiological analyses raise the possibility that calcyon plays a specialized role in regulating activity-dependent removal of synaptic AMPARs. PMID- 19120441 TI - Engineering angiogenesis following spinal cord injury: a coculture of neural progenitor and endothelial cells in a degradable polymer implant leads to an increase in vessel density and formation of the blood-spinal cord barrier. AB - Angiogenesis precedes recovery following spinal cord injury and its extent correlates with neural regeneration, suggesting that angiogenesis may play a role in repair. An important precondition for studying the role of angiogenesis is the ability to induce it in a controlled manner. Previously, we showed that a coculture of endothelial cells (ECs) and neural progenitor cells (NPCs) promoted the formation of stable tubes in vitro and stable, functional vascular networks in vivo in a subcutaneous model. We sought to test whether a similar coculture would lead to the formation of stable functional vessels in the spinal cord following injury. We created microvascular networks in a biodegradable two component implant system and tested the ability of the coculture or controls (lesion control, implant alone, implant + ECs or implant + NPCs) to promote angiogenesis in a rat hemisection model of spinal cord injury. The coculture implant led to a fourfold increase in functional vessels compared with the lesion control, implant alone or implant + NPCs groups and a twofold increase in functional vessels over the implant + ECs group. Furthermore, half of the vessels in the coculture implant exhibited positive staining for the endothelial barrier antigen, a marker for the formation of the blood-spinal cord barrier. No other groups have shown positive staining for the blood-spinal cord barrier in the injury epicenter. This work provides a novel method to induce angiogenesis following spinal cord injury and a foundation for studying its role in repair. PMID- 19120442 TI - AMPA receptors in the medial amygdala are critical for establishing a neuroendocrine memory in the female rat. AB - We sought to examine AMPA receptor (AMPAR) function in the medial posterodorsal amygdala (MePD), as glutamate neurotransmission is critical for the neural response to vaginal-cervical stimulation that initiates pregnancy or pseudopregnancy. Female rats were infused with the AMPAR antagonist CNQX or vehicle directly into the MePD via bilaterally implanted cannulae, then either returned to their homecage (HC), or received 15 mounts-without-intromissions (MO) or 15 intromissions (15I) from a male. Expression of the activity marker EGR-1 was used to determine the CNQX concentration which would prevent mating-induced activation of MePD neurons. Separate cannulated females received CNQX infusions into the MePD prior to receiving 15I, and the oestrous cycle length was monitored by daily vaginal lavages. Infusion of CNQX (500 nm) blocked mating-induced neural activation and lengthened the oestrous cycle, demonstrating AMPAR involvement in the formation of pseudopregnancy. To further explore this involvement, separate groups of 15I, MO and HC females were killed 90 min or 3 h after testing treatment. Brain sections were immunolabeled for AMPAR-subunit GluR1 phosphorylated at one of two sites (Serine-831 or Serine-845), or total GluR1 and GluR2, and immunofluorescence intensity was measured in the MePD, hippocampus and hypothalamus. A mating-induced increase in Serine-831 phosphorylation after 3 h was observed only in the MePD, whereas there was no effect on Serine-845 phosphorylation. Additionally, we observed a time-dependent increase in total GluR1 staining intensity. These results suggest an increased AMPAR function in the MePD after receipt of VCS, and a role for AMPAR in the neural response to VCS resulting in pseudopregnancy. PMID- 19120443 TI - Enrichment and photoperiod interact to affect spatial learning and hippocampal dendritic morphology in white-footed mice (Peromyscus leucopus). AB - In seasonally changing environments, individuals must coordinate endogenous processes with ambient conditions. Winter is a challenging time to survive and reproduce. In order to anticipate decreased food availability and low temperatures in winter, many rodents use decreasing day lengths as a precise temporal cue. Short day lengths alter several adaptations, including reproduction, immune function, aggressive behavior and spatial learning in non tropical rodents. Specifically, short days impair spatial learning in white footed mice (Peromyscus leucopus) and alter dendritic complexity in the hippocampus. The goal of the current study was to determine whether short days constrain neural plasticity. If short days limit the capacity for plasticity, then environmental enrichment, a manipulation that induces morphological changes, should alter dendritic morphology in long, but not short, days. Male white-footed mice were assigned to long (16 : 8 LD) or short (8 : 16 LD) photoperiod in either enriched or standard cages. Enrichment consisted of a large cage, cage mates, Habitrail tubes, a nest box and a running wheel. Mice were tested in the Morris water maze. Reproductive tissues were collected and weighed; brains were processed for dendritic morphology. Short days impaired spatial learning. Short days also reduced spine density on apical dendrites within the CA3 region of the hippocampus. However, enrichment prevented short-day-induced deficits in learning and also increased hippocampal spine density in the CA1 and CA3 regions in short day mice. These results suggest that day length and other non-photic environmental factors interact to regulate dendritic morphology, and that short photoperiods do not constrain the capacity for functional neural plasticity. PMID- 19120445 TI - Perception of global gestalt by temporal integration in simultanagnosia. AB - Patients with bilateral parieto-occipital brain damage may show intact processing of individual objects, while their perception of multiple objects is disturbed at the same time. The deficit is termed 'simultanagnosia' and has been discussed in the context of restricted visual working memory and impaired visuo-spatial attention. Recent observations indicated that the recognition of global shapes can be modulated by the spatial distance between individual objects in patients with simultanagnosia and thus is not an all-or-nothing phenomenon depending on spatial continuity. However, grouping mechanisms not only require the spatial integration of visual information, but also involve integration processes over time. The present study investigated motion-defined integration mechanisms in two patients with simultanagnosia. We applied hierarchical organized stimuli of global objects that consisted of coherently moving dots ('shape-from-motion'). In addition, we tested the patients' ability to recognize biological motion by presenting characteristic human movements ('point-light-walker'). The data revealed largely preserved perception of biological motion, while the perception of motion-defined shapes was impaired. Our findings suggest separate mechanisms underlying the recognition of biological motion and shapes defined by coherently moving dots. They thus argue against a restriction in the overall capacity of visual working memory over time as a general explanation for the impaired global shape recognition in patients with simultanagnosia. PMID- 19120444 TI - Prefrontal and midline interactions mediating behavioural control. AB - Top-down control processes are thought to interact with bottom-up stimulus-driven task demands to facilitate the smooth execution of behaviour. Frontal and midline brain areas in humans are believed to subserve these control processes but their distinct roles and the interactions between them remain to be fully elucidated. In this functional magnetic resonance imaging (fMRI) study, we utilized a GO/NO GO task with cued and uncued inhibitory events to investigate the effect of cue induced levels of top-down control on NO-GO trial response conflict. We found that, on a within-subjects, trial-for-trial basis, high levels of top-down control, as indexed by left dorsolateral prefrontal activation prior to the NO GO, resulted in lower levels of activation on the NO-GO trial in the pre supplementary motor area. These results suggest that prefrontal and midline regions work together to implement cognitive control and reveal that intra subject variability is reflected in these lateral and midline interactions. PMID- 19120446 TI - Characterization of the rice carotenoid cleavage dioxygenase 1 reveals a novel route for geranial biosynthesis. AB - Carotenoid cleavage products--apocarotenoids--include biologically active compounds, such as hormones, pigments and volatiles. Their biosynthesis is initiated by the oxidative cleavage of C-C double bonds in carotenoid backbones, leading to aldehydes and/or ketones. This step is catalyzed by carotenoid oxygenases, which constitute an ubiquitous enzyme family, including the group of plant carotenoid cleavage dioxygenases 1 (CCD1s), which mediates the formation of volatile C(13) ketones, such as beta-ionone, by cleaving the C9-C10 and C9'-C10' double bonds of cyclic and acyclic carotenoids. Recently, it was reported that plant CCD1s also act on the C5-C6/C5'-C6' double bonds of acyclic carotenes, leading to the volatile C(8) ketone 6-methyl-5-hepten-2-one. Using in vitro and in vivo assays, we show here that rice CCD1 converts lycopene into the three different volatiles, pseudoionone, 6-methyl-5-hepten-2-one, and geranial (C(10)), suggesting that the C7-C8/C7'-C8' double bonds of acyclic carotenoid ends constitute a novel cleavage site for the CCD1 plant subfamily. The results were confirmed by HPLC, LC-MS and GC-MS analyses, and further substantiated by in vitro incubations with the monocyclic carotenoid 3-OH-gamma-carotene and with linear synthetic substrates. Bicyclic carotenoids were cleaved, as reported for other plant CCD1s, at the C9-C10 and C9'-C10' double bonds. Our study reveals a novel source for the widely occurring plant volatile geranial, which is the cleavage of noncyclic ends of carotenoids. PMID- 19120447 TI - Arabidopsis thaliana CYP77A4 is the first cytochrome P450 able to catalyze the epoxidation of free fatty acids in plants. AB - An approach based on an in silico analysis predicted that CYP77A4, a cytochrome P450 that so far has no identified function, might be a fatty acid-metabolizing enzyme. CYP77A4 was heterologously expressed in a Saccharomyces cerevisiae strain (WAT11) engineered for cytochrome P450 expression. Lauric acid (C(12:0)) was converted into a mixture of hydroxylauric acids when incubated with microsomes from yeast expressing CYP77A4. A variety of physiological C(18) fatty acids were tested as potential substrates. Oleic acid (cis-Delta(9)C(18:1)) was converted into a mixture of omega-4- to omega-7-hydroxyoleic acids (75%) and 9,10 epoxystearic acid (25%). Linoleic acid (cis,cis-Delta(9),Delta(12)C(18:2)) was exclusively converted into 12,13-epoxyoctadeca-9-enoic acid, which was then converted into diepoxide after epoxidation of the Delta(9) unsaturation. Chiral analysis showed that 9,10-epoxystearic acid was a mixture of 9S/10R and 9R/10S in the ratio 33 : 77, whereas 12,13-epoxyoctadeca-9-enoic acid presented a strong enantiomeric excess in favor of 12S/13R, which represented 90% of the epoxide. Neither stearic acid (C(18:0)) nor linolelaidic acid (trans,trans Delta(9),Delta(12)C(18:2)) was metabolized, showing that CYP77A4 requires a double bond, in the cis configuration, to metabolize C(18) fatty acids. CYP77A4 was also able to catalyze the in vitro formation of the three mono-epoxides of alpha-linolenic acid (cis,cis,cis-Delta(9),Delta(12),Delta(15)C(18:3)), previously described as antifungal compounds. Epoxides generated by CYP77A4 are further metabolized to the corresponding diols by epoxide hydrolases located in microsomal and cytosolic subcellular fractions from Arabidopsis thaliana. The concerted action of CYP77A4 with epoxide hydrolases and hydroxylases allows the production of compounds involved in plant-pathogen interactions, suggesting a possible role for CYP77A4 in plant defense. PMID- 19120448 TI - Amino acid limitation regulates the expression of genes involved in several specific biological processes through GCN2-dependent and GCN2-independent pathways. AB - Evidence has accumulated that amino acids play an important role in controlling gene expression. Nevertheless, two components of the amino acid control of gene expression are not yet completely understood in mammals: (a) the target genes and biological processes regulated by amino acid availability, and (b) the signaling pathways that mediate the amino acid response. Using large-scale analysis of gene expression, the objective of this study was to gain a better understanding of the control of gene expression by amino acid limitation. We found that a 6 h period of leucine starvation regulated the expression of a specific set of genes: 420 genes were up-regulated by more than 1.8-fold and 311 genes were down-regulated. These genes were involved in the control of several biological processes, such as amino acid metabolism, lipid metabolism and signal regulation. Using GCN2-/- cells and rapamycin treatment, we checked for the role of mGCN2 and mTORC1 kinases in this regulation. We found that (a) the GCN2 pathway was the major, but not unique, signaling pathway involved in the up- and down-regulation of gene expression in response to amino acid starvation, and (b) that rapamycin regulates the expression of a set of genes that only partially overlaps with the set of genes regulated by leucine starvation. PMID- 19120449 TI - Destabilization of psychrotrophic RNase HI in a localized fashion as revealed by mutational and X-ray crystallographic analyses. AB - The Arg97 --> Gly and Asp136 --> His mutations stabilized So-RNase HI from the psychrotrophic bacterium Shewanella oneidensis MR-1 by 5.4 and 9.7 degrees C, respectively, in T(m), and 3.5 and 6.1 kJ x mol(-1), respectively, in DeltaG(H2O). These mutations also stabilized the So-RNase HI derivative (4x-RNase HI) with quadruple thermostabilizing mutations in an additive manner. As a result, the resultant sextuple mutant protein (6x-RNase HI) was more stable than the wild-type protein by 28.8 degrees C in T(m) and 27.0 kJ x mol(-1) in DeltaG(H2O). To analyse the effects of the mutations on the protein structure, the crystal structure of the 6x-RNase HI protein was determined at 2.5 A resolution. The main chain fold and interactions of the side-chains of the 6x RNase HI protein were basically identical to those of the wild-type protein, except for the mutation sites. These results indicate that all six mutations independently affect the protein structure, and are consistent with the fact that the thermostabilizing effects of the mutations are roughly additive. The introduction of favourable interactions and the elimination of unfavourable interactions by the mutations contribute to the stabilization of the 6x-RNase HI protein. We propose that So-RNase HI is destabilized when compared with its mesophilic and thermophilic counterparts in a localized fashion by increasing the number of amino acid residues unfavourable for protein stability. PMID- 19120450 TI - Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced chemokine release in both TRAIL-resistant and TRAIL-sensitive cells via nuclear factor kappa B. AB - Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in a variety of tumour cells, but not in most normal cells, and has attracted considerable attention for its potential use in cancer therapy. Recently, increasing evidence has shown that TRAIL is involved in inflammation, although much of this evidence is controversial. In this article, it is shown that TRAIL induces CXCL2, CCL4 and CCL20 secretion in a nuclear factor kappa B dependent manner. The dominant negative constructs of tumour necrosis factor receptor-associated death domain protein (TRADD) and tumour necrosis factor receptor-associated factor 2 are unable to block TRAIL-induced chemokine up regulation, and the dominant negative construct of TRADD may even enhance TRAIL triggered signals. Using small interfering RNA, receptor interacting protein has been demonstrated to be essential for TRAIL-induced chemokine release. Furthermore, it has been demonstrated that p38 mitogen-activated protein kinase is involved in TRAIL-induced chemokine release without any effects on nuclear factor kappa B activation, suggesting that some unknown transcription factors may be activated by TRAIL. Using a xenograft tumour model, it has been illustrated that TRAIL can also induce chemokine release in vivo. Although these chemokines induced by TRAIL are inflammatory chemokines, their functions are not restricted to inflammation and require further examination. Our results indicate that attention should be paid to the side-effects of TRAIL treatment, not only in TRAIL-resistant but also in TRAIL-sensitive tumour cells. PMID- 19120452 TI - Coq10, a mitochondrial coenzyme Q binding protein, is required for proper respiration in Schizosaccharomyces pombe. AB - It has been widely accepted that most coenzyme Q (CoQ) exists freely in the mitochondrial membrane as a CoQ pool. However, the recent identification of a mitochondrial CoQ-binding protein, termed Coq10, in budding yeast has the potential to change our current view of CoQ status in membranes. Here, we studied the counterpart of budding yeast Coq10 (also termed Coq10) in fission yeast. Fission yeast coq10 null mutants exhibited a similar, but less severe, phenotype to CoQ-deficient fission yeast, including the requirement for antioxidants for proper growth on minimal medium, increased sensitivity to H(2)O(2), high levels of H(2)S production, and a deficiency in respiration. The coq10 null mutant produced nearly normal levels of CoQ10, suggesting that coq10 does not belong to the group of CoQ biosynthetic genes. To elucidate the role of Coq10, we expressed recombinant coq10 in Escherichia coli, and found that CoQ8 was present in purified recombinant Coq10. Mutational analysis of 13 conserved residues of Coq10 revealed that two hydrophobic amino acid residues, leucine 63 (L63) and tryptophan 104 (W104), play an important role in Coq10 binding to CoQ. An L63A/W104A double mutant of Coq10 exhibited lower CoQ-binding activity than either of the single mutants, and was unable to complement the coq10 deletion in fission yeast. A human Coq10 ortholog was able to functionally compensate for the absence of coq10 in fission yeast, suggesting that Coq10 is important for proper respiration in a variety of organisms. PMID- 19120451 TI - Characterization of a monoclonal antibody as the first specific inhibitor of human NTP diphosphohydrolase-3 : partial characterization of the inhibitory epitope and potential applications. AB - The study and therapeutic modulation of purinergic signaling is hindered by a lack of specific inhibitors for NTP diphosphohydrolases (NTPDases),which are the terminating enzymes for these processes. In addition, little is known of the NTPDase protein structural elements that affect enzymatic activity and which could be used as targets for inhibitor design. In the present study, we report the first inhibitory monoclonal antibodies specific for an NTPDase, namely human NTPDase3 (EC 3.6.1.5), as assessed by ELISA, western blotting, flow cytometry, immunohistochemistry and inhibition assays. Antibody recognition of NTPDase3 is greatly attenuated by denaturation with SDS, and abolished by reducing agents, indicating the significance of the native conformation and the disulfide bonds for epitope recognition. Using site-directed chemical cleavage, the SDS-resistant parts of the epitope were located in two fragments of the C-terminal lobe ofNTPDase3 (i.e. Leu220-Cys347 and Cys347-Pro485), which are both required for antibody binding. Additional site-directed mutagenesis revealed the importance of Ser297 and the fifth disulfide bond (Cys399-Cys422) for antibody binding, indicating that the discontinuous inhibitory epitope is located on the extracellular C-terminal lobe of NTPDase3. These antibodies inhibit recombinant NTPDase3 by 60-90%, depending on the conditions. More importantly, they also efficiently inhibit the NTPDase3expressed in insulin secreting human pancreatic islet cells in situ. Because insulin secretion is modulated by extracellular ATP and purinergic receptors, this finding suggests the potential application of these inhibitory antibodies for the study and control of insulin secretion. PMID- 19120453 TI - Dimerization of the yeast eukaryotic translation initiation factor 5A requires hypusine and is RNA dependent. AB - Post-translational modification of the highly conserved K51 residue of the Saccharomyces cerevisiae eukaryotic translation initiation factor 5A (eIF5A) to form hypusine, is essential for its many functions including the binding of specific mRNAs. We characterized hypusinated yeast eIF5A by size-exclusion chromatography and native PAGE, showing that the protein exists as a homodimer. A K51R mutant, which was not functional in vivo eluted as a monomer and inhibition of hypusination abolished dimerization. Furthermore, treatment of dimeric eIF5A with RNase A resulted in disruption of the dimer, leading us to conclude that RNA binding is also required for dimerization of eIF5A. We present a model of dimerization, based on the Neurospora crassa structural analogue, HEX-1. PMID- 19120454 TI - Regulators of G-protein signalling are modulated by bacterial lipopeptides and lipopolysaccharide. AB - Regulators of G-protein signalling accelerate the GTPase activity of G(alpha) subunits, driving G proteins in their inactive GDP-bound form. This property defines them as GTPase activating proteins. Here the effect of different Toll like receptor agonists on RGS1 and RGS2 expression in murine bone marrow-derived macrophages and J774 cells was analysed. After stimulation with TLR2/1 or TLR2/6 lipopeptide ligands and the TLR4/MD2 ligand lipopolysaccharide, microarray analyses show only modulation of RGS1 and RGS2 among all the regulators of G protein signalling tested. Real-time PCR confirmed modulation of RGS1 and RGS2. In contrast to RGS2, which was always downregulated, RGS1 mRNA was upregulated during the first 30 min after stimulation, followed by downregulation. Similar results were also found in the murine macrophage cell line J774. The ligand for intracellular TLR9 modulates RGS1 and RGS2 in a similar manner. However, the TLR3 ligand poly(I:C) permanently upregulates RGS1 and RGS2 expression indicating a different modulation by the MyD88- and TRIF-signalling pathway. This was confirmed using MyD88(-/-) and TRIF(-/-) bone marrow-derived macrophages. Modulation of RGS1 and RGS2 by Toll-like receptor ligands plays an important role during inflammatory and immunological reactions after bacterial and viral infection. PMID- 19120455 TI - Competition between innate multidrug resistance and intracellular binding of rhodamine dyes. AB - The present study aimed to elucidate the contribution of the intracellular binding of drugs to multidrug resistance. For this purpose, uptake of rhodamines was studied in cells whose mitochondria had been uncoupled with carbonyl cyanide m-chlorophenylhydrazone. Surprisingly, in a variety of drug-untreated cells, presumed to be sensitive to multidrug resistance-type drugs, rhodamines were excluded from entering the cells. Thus, the amount of rhodamine 123 taken up into parental untreated K562 cells was less than the amount bound to the cell exterior. Rhodamine uptake was prevented by an active efflux pump. The efflux was inhibited by 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) and MK571 and, to a lesser extent, by ATP depletion, indomethacin, probenecid and vanadate. All the inhibitors, apart from NBD-Cl, are known to modulate multidrug resistance associated protein (MRP) 1. Because MRP1 was expressed in all the cell lines tested and the efflux of rhodamines in MRP1 over-expressing cells was abolished by NBD-Cl, it appears that rhodamines are excluded from these cells by MRP1. On the other hand, the uptake of rhodamines into cells respiring with their coupled mitochondria demonstrated diminished sensitivity to NBD-Cl and MK571. Thus, active pumping into the mitochondria allowed enhanced uptake into the cells, overcoming the innate resistance. The innate resistance provided by MRP1 to cells prevents rhodamine dyes, and possibly drugs such as doxorubicin, from achieving equilibration of their concentration in the cytoplasm with their concentration in the external medium. The protection provided to multidrug resistance cells by ABC transporters has to overcome competition by passive uptake of the drugs and binding/uptake of the drugs into intracellular targets. PMID- 19120456 TI - Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology. AB - Quantitative PCR (Q-PCR or real-time PCR) approaches are now widely applied in microbial ecology to quantify the abundance and expression of taxonomic and functional gene markers within the environment. Q-PCR-based analyses combine 'traditional' end-point detection PCR with fluorescent detection technologies to record the accumulation of amplicons in 'real time' during each cycle of the PCR amplification. By detection of amplicons during the early exponential phase of the PCR, this enables the quantification of gene (or transcript) numbers when these are proportional to the starting template concentration. When Q-PCR is coupled with a preceding reverse transcription reaction, it can be used to quantify gene expression (RT-Q-PCR). This review firstly addresses the theoretical and practical implementation of Q-PCR and RT-Q-PCR protocols in microbial ecology, highlighting key experimental considerations. Secondly, we review the applications of (RT)-Q-PCR analyses in environmental microbiology and evaluate the contribution and advances gained from such approaches. Finally, we conclude by offering future perspectives on the application of (RT)-Q-PCR in furthering understanding in microbial ecology, in particular, when coupled with other molecular approaches and more traditional investigations of environmental systems. PMID- 19120457 TI - Spatial comparison of total vs. active bacterial populations by coupling genetic fingerprinting and clone library analyses in the NW Mediterranean Sea. AB - Spatial distributions of both total (i.e. 16S rDNA-based fingerprints) and active (i.e. 16S rRNA-based fingerprints) bacterial populations, together with total bacterial activity measured by 3H-leucine incorporation, were studied along a 98 km transect in the NW Mediterranean Sea. Capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) fingerprinting was coupled to a clone library, allowing CE-SSCP peaks identification and the monitoring of the spatial variation of bacterial phylotypes. Up to 80% of the community peaks matched those obtained from clone library sequences, accounting for 86.7% of the total fingerprinting area. A good agreement was found between the relative abundance of Prochlorococcus in the CE-SSCP fingerprints and flow cytometry counts (r2=0.66, P<0.05). The largest differences between total and active bacterial populations distribution were found at depths with higher bacterial activity (i.e. surface and deep chlorophyll maximum, DCM). SAR11 at the surface and Gammaproteobacteria at the DCM were the most abundant groups on the 16S rDNA-based fingerprints. However, their ratio of relative importance between rRNA : rDNA was <1 in most cases. Conversely, ratios observed for Prochlorococcus, were consistently >1 both at the surface and at the DCM. These results emphasize the need for combining rDNA- and rRNA-based analyses to better understand the functional role of individual bacterial populations in situ. PMID- 19120458 TI - Single-cell vs. bulk activity properties of coastal bacterioplankton over an annual cycle in a temperate ecosystem. AB - The connections between single-cell activity properties of heterotrophic planktonic bacteria and whole community metabolism are still poorly understood. Here, we show flow cytometry single-cell analysis of membrane-intact (live), high nucleic acid (HNA) content and actively respiring (CTC+) bacteria with samples collected monthly during 2006 in northern Spain coastal waters. Bulk activity was assessed by measuring 3H-Leucine incorporation and specific growth rates. Consistently, different single-cell relative abundances were found, with 60-100% for live, 30-84% for HNA and 0.2-12% for CTC+ cells. Leucine incorporation rates (2-153 pmol L(-1) h(-1)), specific growth rates (0.01-0.29 day(-1)) and the total and relative abundances of the three single-cell groups showed marked seasonal patterns. Distinct depth distributions during summer stratification and different relations with temperature, chlorophyll and bacterial biovolume suggest the existence of different controlling factors on each single-cell property. Pooled leucine incorporation rates were similarly correlated with the abundance of all physiological groups, while specific growth rates were only substantially explained by the percentage of CTC+ cells. However, the ability to reduce CTC proved notably better than the other two single-cell properties at predicting bacterial bulk rates within seasons, suggesting a tight linkage between bacterial individual respiration and biomass production at the community level. PMID- 19120459 TI - Influence of nutrient status and grazing pressure on the fate of Francisella tularensis in lake water. AB - The natural reservoir of Francisella tularensis, the causative agent of tularaemia, is yet to be identified. We investigated the possibility that Francisella persists in natural aquatic ecosystems between outbreaks. It was hypothesized that nutrient-rich environments, with strong protozoan predation, favour the occurrence of the tularaemia bacterium. To investigate the differences in adaptation to aquatic environments of the species and subspecies of Francisella, we screened 23 strains for their ability to survive grazing by the ciliate Tetrahymena pyriformis. All the Francisella strains tested were consumed at a low rate, although significant differences between subspecies were found. The survival and virulence of gfp-labelled F. tularensis ssp. holarctica were then studied in a microcosm experiment using natural lake water, with varying food web complexities and nutrient availabilities. High nutrient conditions in combination with high abundances of nanoflagellates were found to favour F. tularensis ssp. holarctica. The bacterium was observed both free-living and within the cells of a nanoflagellate. Francisella tularensis entered a viable but nonculturable state during the microcosm experiment. When studied over a longer period of time, F. tularensis ssp. holarctica survived in the lake water, but loss of virulence was not prevented by either high nutrient availability or the presence of predators. PMID- 19120460 TI - Arbuscular mycorrhizal dynamics in a chronosequence of Caragana korshinskii plantations. AB - Arbuscular mycorrhizal (AM) fungi in a chronosequence of 5-42-year-old Caragana korshinskii plantations in the semi-arid Loess Plateau region of northwestern China were investigated. AM fungi colonization, spore diversity and PCR-denatured gradient gel electrophoresis-based AM fungal SSU rRNA gene sequences were analyzed. AM fungi colonization [measured as the percent of root length (%RLC), vesicular (%VC) and arbuscular (%AC) colonization] and spore density were significantly correlated with sampling month, but not with plant age, except for %RLC. The percent of vesicular colonization was negatively correlated with soil total nitrogen and organic carbon, and spore density was negatively correlated with soil moisture and available phosphorus. Ten distinguishable AM fungal spore morphotypes, nine Glomus and one Scutellospora species, were found. Nine AM fungal Glomus phylotypes were identified by sequencing, but at each sampling time only four to six AM fungal phylotypes were detected. The AM fungal community was significantly seasonal, whereas the AM fungal species richness did not increase with plantation age. A significant change in AM fungal colonization and community composition over an annual cycle was observed in this study, and our results suggest that the changes of AM are the product of the interaction between host phenology, soil characteristics and habitat. Understanding these interactions is essential if habitat restoration is to be effective. PMID- 19120461 TI - Rhizobia with different symbiotic efficiencies nodulate Acaciella angustissima in Mexico, including Sinorhizobium chiapanecum sp. nov. which has common symbiotic genes with Sinorhizobium mexicanum. AB - Bacteria from nodules of the legume Acaciella angustissima native to the south of Mexico were characterized genetically and their nodulation and competitiveness were evaluated. Phylogenetic studies derived from rpoB gene sequences indicated that A. angustissima is nodulated by Sinorhizobium mexicanum, Rhizobium tropici, Mesorhizobium plurifarium and Agrobacterium tumefaciens and by bacteria related to Sinorhizobium americanum, Sinorhizobium terangae, Rhizobium etli and Rhizobium gallicum. A new lineage related to S. terangae is recognized based on the sequences of gyrA, nolR, recA, rpoB and rrs genes, DNA-DNA hybridization and phenotypic characteristics. The name for this new species is Sinorhizobium chiapanecum and its type strain is ITTG S70T. The symbiotic genes nodA and nifH were similar to those from S. mexicanum strains, which are Acaciella symbionts as well, with nodA gene sequences grouped within a cluster of nod genes from strains that nodulate plants from the Mimosoideae subfamily of the Leguminosae. Sinorhizobium isolates were the most frequently obtained from A. angustissima nodules and were among the best strains to promote plant growth in A. angustissima and to compete in interstrain nodule competition assays. Lateral transfer of symbiotic genes is not evident among the genera that nodulate A. angustissima (Rhizobium, Sinorhizobium and Mesorhizobium) but may occur among the sympatric and closely related sinorhizobia that nodulate Acaciella. PMID- 19120462 TI - An initial characterization of the mercury resistance (mer) system of the thermophilic bacterium Thermus thermophilus HB27. AB - The evolutionary origin of the broadly distributed mer system, which plays an important role in mercury detoxification and biogeochemistry, is presently unknown. The phylum Deinococcus/Thermus was found to be one of the deepest branching bacterial lineage to have a homolog of merA, which specifies reduction of ionic to elemental mercury, and the mercuric reductase (MerA) of Thermus thermophilus HB27 was found to be basal to all bacterial MerA when this protein's phylogeny was constructed. A merA mutant of HB27 was fourfolds more sensitive to mercury toxicity than the wild type (wt), and lost detectable MerA-specific activities. The merA gene in HB27 was transcribed on a polycistronic message downstream from ORF encoding for homologs of O-acetyl-l-homoserine/O-acetyl serine (OAH/OAS) sulfhydrylase and MerR, the mer operon transcription regulator, from a promoter located 69 nucleotides upstream of the sulfhydrylase translation start codon. The transcription of the putative mer operon in HB27 was induced 66.8+/-15.8-fold by exposure to 1 muM HgCl2. The optimal temperature for MerA specific activity corresponded to this strain's optimal growth temperature, 70 degrees C. Thus, T. thermophilus is the earliest mercury-resistant bacterium identified to date, a finding consistent with the hypothesis that the mer system originated among thermophilic microorganisms from geothermal environments. PMID- 19120463 TI - Sulfonate desulfurization in Rhodococcus from wheat rhizosphere communities. AB - Organically bound sulfur makes up about 90% of the total sulfur in soils, with sulfonates often the dominant fraction. Actinobacteria affiliated to the genus Rhodococcus were able to desulfonate arylsulfonates in wheat rhizospheres from the Broadbalk long-term field wheat experiment, which includes plots treated with inorganic fertilizer with and without sulfate, with farmyard manure, and unfertilized plots. Direct isolation of desulfonating rhizobacteria yielded Rhodococcus strains which grew well with a range of sulfonates, and contained the asfAB genes, known to be involved in sulfonate desulfurization by bacteria. Expression of asfA in vitro increased >100-fold during growth of the Rhodococcus isolates with toluenesulfonate as sulfur source, compared with growth with sulfate. By contrast, the closely related Rhodococcus erythropolis and Rhodococcus opacus type strains had no desulfonating activity and did not contain asfA homologues. The overall actinobacterial community structure in wheat rhizospheres was influenced by the sulfur fertilization regime, as shown by specific denaturing gradient gel electrophoresis of PCR amplified 16S rRNA gene fragments, and asfAB clone library analysis identified nine different asfAB genotypes closely affiliated to the Rhodococcus isolates. However, asfAB-based multiplex restriction fragment length polymorphism (RFLP)/terminal-RFLP analysis of wheat rhizosphere communities revealed only slight differences between the fertilization regimes, suggesting that the desulfonating Rhodococcus community does not specifically respond to changes in sulfate supply. PMID- 19120464 TI - Nitrate-reducing, sulfide-oxidizing bacteria as microbial oxidants for rapid biological sulfide removal. AB - The emission of hydrogen sulfide into the atmosphere of sewer systems induces the biological production of sulfuric acid, causing severe concrete corrosion. As a possible preventive solution, a microbial consortium of nitrate-reducing, sulfide oxidizing bacteria (NR-SOB) was enriched in a continuously stirred tank reactor in order to develop a biological technique for the removal of dissolved sulfide. The consortium, dominated by Arcobacter sp., was capable of removing 99% of sulfide. Stable isotope fractioning of the sulfide indicated that the oxidation was a biological process. The capacity of the NR-SOB consortium for rapid removal of sulfide was demonstrated by using it as an inoculum in synthetic and real sewage. Removal rates up to 52 mg sulfide-S g VSS(-1) h(-1) were achieved, to our knowledge the highest removal rate reported so far for freshwater species in the absence of molecular oxygen. Further long-term incubation experiments revealed the capacity of the bacteria to oxidize sulfide without the presence of nitrate, suggesting that an oxidized redox reserve is present in the culture. PMID- 19120465 TI - Quantitative analysis of cellulose degradation and growth of cellulolytic bacteria in the rumen. AB - Ruminant animals digest cellulose via a symbiotic relationship with ruminal microorganisms. Because feedstuffs only remain in the rumen for a short time, the rate of cellulose digestion must be very rapid. This speed is facilitated by rumination, a process that returns food to the mouth to be rechewed. By decreasing particle size, the cellulose surface area can be increased by up to 10(6)-fold. The amount of cellulose digested is then a function of two competing rates, namely the digestion rate (K(d)) and the rate of passage of solids from the rumen (K(p)). Estimation of bacterial growth on cellulose is complicated by several factors: (1) energy must be expended for maintenance and growth of the cells, (2) only adherent cells are capable of degrading cellulose and (3) adherent cells can provide nonadherent cells with cellodextrins. Additionally, when ruminants are fed large amounts of cereal grain along with fiber, ruminal pH can decrease to a point where cellulolytic bacteria no longer grow. A dynamic model based on STELLA software is presented. This model evaluates all of the major aspects of ruminal cellulose degradation: (1) ingestion, digestion and passage of feed particles, (2) maintenance and growth of cellulolytic bacteria and (3) pH effects. PMID- 19120466 TI - High genetic similarity between two geographically distinct strains of the sulfur oxidizing symbiont 'Candidatus Thiobios zoothamnicoli'. AB - The giant marine ciliate Zoothamnium niveum (Ciliophora, Oligohymenophora) is obligatorily covered by a monolayer of putative chemoautotrophic sulfur-oxidizing (thiotrophic) bacteria. For Z. niveum specimens from the Caribbean Sea it has been demonstrated that this ectosymbiotic population consists of only a single pleomorphic phylotype described as Candidatus Thiobios zoothamnicoli. The goal of our study was to identify and phylogenetically analyse the ectosymbiont(s) of a recently discovered Z. niveum population from the Mediterranean Sea, and to compare marker genes encoding key enzymes of the carbon and sulfur metabolism between the two symbiont populations. We identified a single bacterial phylotype representing the ectosymbiont of Z. niveum from the Mediterranean population showing 99.7% 16S rRNA gene (99.2% intergenic spacer region) similarity to the Caribbean Z. niveum ectosymbiont. Genes encoding enzymes typical for an inorganic carbon metabolism [ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO)] and for sulfur metabolism (5'-adenylylsulfate reductase, dissimilatory sulfite reductase) were detected in both symbiotic populations. The very high amino acid sequence identity (97-100%) and the high nucleic acid sequence identity (90-98%) of these marker enzymes in two geographically distant symbiont populations suggests that the association of Z. niveum with Cand. Thiobios zoothamnicoli is very specific as well as temporally and spatially stable. PMID- 19120467 TI - Lack of endosymbiont release by two Lucinidae (Bivalvia) of the genus Codakia: consequences for symbiotic relationships. AB - Associations between marine invertebrates and chemoautotrophic bacteria constitute a wide field for the study of symbiotic associations. In these interactions, symbiont transmission must represent the cornerstone allowing the persistence of the association throughout generations. Within Bivalvia, in families such as Solemyidae or Vesicomyidae, symbiont transmission is undoubtedly vertical. However, in Lucinidae, symbiont transmission is described in the literature as 'environmental', symbionts being acquired from the environment by the new host generations. Hence, if there is transmission, symbionts should be transmitted from adults to juveniles via the environment. Consequently, we should observe a release of the symbiont by adults. We attempted to detect such a release within two Lucinidae species of the genus Codakia. We sampled 10 Codakia orbicularis and 20 Codakia orbiculata distributed in 10 crystallizing dishes containing filtered seawater. During 1 month of investigation, we analyzed water of the dishes in order to detect any release of a symbiont using catalyzed report deposition-FISH techniques. For 140 observations realized during this period, we did not observe any release of symbionts. This suggests that the idea of host-to host passage in Lucinidae is inaccurate. We could therefore consider that the transmission mode from generation to generation does not occur within Lucinidae, symbiosis appearing to be advantageous in this case only for the host, and constitutes an evolutionary dead-end for the bacteria. PMID- 19120468 TI - Annual growth layers as proxies of past growth conditions for benthic microbial mats in a perennially ice-covered Antarctic lake. AB - Perennial microbial mats can be the dominant autotrophic community in Antarctic lakes. Their seasonal growth results in clearly discernible annual growth layering. We examined features of live microbial mats from a range of depths in Lake Hoare, Antarctica, that are likely to be preserved in these layers to determine their potential as proxies of past growth performance. Cyanobacteria dominated the mat for all but the deepest depth sampled. Changes in areal concentrations of phycobilin pigments, organic matter and extracellular polysaccharide and in species composition did not correspond to changes in various water column properties, but showed a linear relationship with irradiance. Carbonate accumulation in the mats correlated with biomass markers and may be inferred as an index of mat performance. We examined the carbonate content of annual layers laid down from 1958-1959 to 1994-1995 in sediment cores from 12 m depth. The carbonate content in the layer showed a significant correlation with the mean summer air temperature. These data suggest a link between air temperature and microbial mat growth performance, and suggest that it is mediated via irradiance. Laminated microbial mats in Antarctic lakes have the potential to act as fine-resolution records of environmental conditions in the recent past, although interpretation is complex. PMID- 19120469 TI - Evaluation of taxa-specific real-time PCR, whole-cell FISH and morphotaxonomy analyses for the detection and quantification of the toxic microalgae Alexandrium minutum (Dinophyceae), Global Clade ribotype. AB - The dinoflagellate genus Alexandrium contains neurotoxin-producing species that have adversely affected the aquaculture industry in many countries. The morphological similarity between Alexandrium species has led to the development of molecular methods for the discrimination, enumeration and monitoring of toxic and nontoxic species. A quantitative real-time PCR assay (qRT-PCR) targeting the internal transcribed spacer 1-5.8S rRNA gene using hybridization probe technology was developed for the potentially toxic species Alexandrium minutum (Global Clade) (GC). The assay was specific with a detection limit of less than one cell equivalent. The assay was used to detect and quantify A. minutum (GC) in seawater samples collected during summer 2007 in Cork Harbour, Ireland. The results were compared with those obtained using whole-cell FISH (WC-FISH) and morphotaxonomy analyses. Alexandrium minutum did not reach high bloom concentrations over the sampling period (maximum of c. 6 x 10(4) cells L(-1)), and the average concentrations determined using qRT-PCR, WC-FISH and morphotaxonomy did not significantly differ in eight of nine comparisons. Regression curves showed positive relationships between the methods; WC-FISH and qRT-PCR slightly under- and overestimated, respectively, the A. minutum concentrations compared with the morphotaxonomy method. The qRT-PCR assay for A. minutum (GC) offers high throughput sample analysis and may prove suitable for implementation in microalgae monitoring programmes and assist in population dynamics studies of the species. PMID- 19120470 TI - Innate immunity to pathogens: diversity in receptors for microbial recognition. PMID- 19120471 TI - The long pentraxin PTX3 as a prototypic humoral pattern recognition receptor: interplay with cellular innate immunity. AB - The innate immune system consists of a cellular arm and a humoral arm. Components of humoral immunity include diverse molecular families, which represent functional ancestors of antibodies. They play a key role as effectors and modulators of innate resistance in animals and humans, interacting with cellular innate immunity. The prototypic long pentraxin, pentraxin 3 (PTX3), represents a case in point of this interplay. Gene targeting of this evolutionarily conserved long pentraxin has unequivocally defined its role at the crossroads of innate immunity, inflammation, matrix deposition, and female fertility. Phagocytes represent a key source of this fluid-phase pattern recognition receptor, which, in turn, facilitates microbial recognition by phagocytes acting as an opsonin. Moreover, PTX3 has modulatory functions on innate immunity and inflammation. Here, we review the studies on PTX3 which emphasize the complexity and complementarity of the crosstalk between the cellular and humoral arms of innate immunity. PMID- 19120472 TI - Conserved domains of the class A scavenger receptors: evolution and function. AB - The class A scavenger receptors are phagocytic pattern recognition receptors that are well represented in vertebrate genomes. The high level of conservation among vertebrates implies that this is an evolutionarily conserved family of receptors and indicates the presence of a common ancestral gene. The identity of this ancestral gene is not clear, as it appears that many of the domains of the scavenger receptors (e.g. collagenous, scavenger receptor cysteine rich) originated early in evolutionary history and are found in many combinations, often in genes of unknown function. These early receptors may function in cell cell recognition, aggregation, or lipid recognition, and their involvement in pattern recognition, phagocytosis, and homeostasis may have been adaptations of such conserved patterns. Herein, we reclassify the class A scavenger receptors based on recent discoveries of new members of this family, describe the evolution of the various domains of the class A scavenger receptors, and discuss the appearance and function of these domains through evolutionary history. PMID- 19120473 TI - Regulatory molecules required for nucleotide-sensing Toll-like receptors. AB - Toll-like receptors (TLRs) play an important role in innate immune responses against bacteria and viruses. TLRs localize either on the cell surface or in intracellular vesicular compartments. The cell-surface TLRs, including TLR1, TLR2, TLR4, and TLR6, recognize microbial membrane lipids, whereas TLR3, TLR7, TLR8, and TLR9 recognize pathogen-derived nucleotides in intracellular compartments. TLR7 and TLR9 respond to host-derived nucleotides as well, and they have been implicated in a variety of autoimmune diseases. Safety mechanisms are required to avoid detrimental autoimmune responses. TLR7 and TLR9 are sequestered in the endoplasmic reticulum (ER) in a resting state and traffic to endolysosomes upon ligand-induced stimulation. Sequestration in the ER is a mechanism controlling TLR7/9 responses. A chaperone, gp96, in the ER is reported to regulate TLR7/9 maturation. gp96 is associated with TLR9 and is required for ligand-induced activation of TLR7/9. Two molecules in the ER are reported to regulate TLR7/9 trafficking to endolysosomes. PRAT4A (a protein associated with TLR4 A) is associated with TLR9 and is required for ligand-induced trafficking of TLR9 to endolysosomes. UNC93B1 is specifically associated with TLR3, TLR7, TLR9, and TLR13 and regulates ligand-induced trafficking of TLR7 and TLR9 from the ER to endolysosomes. These molecules are potential therapeutic targets for controlling dysregulated TLR7/9 responses in autoimmune diseases. PMID- 19120474 TI - Functional evolution of the TICAM-1 pathway for extrinsic RNA sensing. AB - The type I interferon (IFN) is a host defense factor against microbial pathogens in vertebrates. In mammals, retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5) in the cytoplasm are regarded as sensors for double-stranded RNA (dsRNA) and trigger IFN regulatory factor-3 (IRF-3) activation followed by type I IFN induction through the mitochondrial antiviral signaling (MAVS) adapter. This intrinsic pathway appears to link the main protective responses against RNA virus infection in mammals. On the other hand, human Toll-like receptor 3 (TLR3) is localized in the endosomal membrane or cell surface and signals the presence of extrinsic dsRNA. In response to RNA stimulation, TLR3 recruits the Toll-interleukin 1 receptor domain (TIR) containing adapter molecule 1 (TICAM-1) adapter and induces IRF-3 activation followed by IFN-beta promoter activation. Human TLR3 is localized limitedly extent in myeloid dendritic cells, fibroblasts, and epithelial cells. The TICAM-1 and cytoplasmic MAVS pathways converge at the IRF-3-activating kinase in human cells. The reason for the involvement of this extrinsic mode of IFN-inducing pathways in the dsRNA response remains unknown. In fish, two TLRs, i.e. endoplasmic TLR3 and cell surface TLR22, participate in teleost IFN production without the activation of IRF-3. TLR22 is distinct from mammalian TLR3 in terms of cellular localization, ligand selection, and tissue distribution. TLR22 may be a functional substitute for human cell surface TLR3 and may serve as a surveillance molecule for detecting dsRNA virus infection and alerting the immune system for antiviral protection in fish. In this review, we discuss the fundamentals of the extrinsic dsRNA recognition system, which has evolved to induce cellular effectors to cope with dsRNA virus infection across different vertebrate species. PMID- 19120475 TI - RNA recognition and signal transduction by RIG-I-like receptors. AB - Viral infection is detected by cellular sensor molecules as foreign nucleic acids and initiates innate antiviral responses, including the activation of proinflammatory cytokines and type I interferon (IFN). Recent identification of cytoplasmic viral sensors, such as retinoic acid-inducible gene-I-like receptors (RLRs), highlights their significance in the induction of antiviral innate immunity. Moreover, it is intriguing to understand how they can discriminate endogenous RNA from foreign viral RNA and initiate signaling cascades leading to the induction of type I IFNs. This review focuses on the current understanding of the molecular machinery underlying RNA recognition and subsequent signal transduction by RLRs. PMID- 19120476 TI - Approaching the RNA ligand for RIG-I? AB - Innate and antigen-specific antiviral immunity are triggered by immunorecognition of viral nucleic acids. The helicase retinoic acid-inducible gene I (RIG-I) (also known as DDX58) is the key sensor of negative strand RNA viruses in the cytosol of cells. RNA containing a triphosphate at the 5'-end was shown to activate RIG I, but the exact structure of RNA supporting 5'-triphosphate recognition, the requirement of a 5'-triphosphate group, as well as the existence of RNA structures detected by RIG-I in the absence of 5'-triphosphate remain controversial. Here, we revisit the literature on RIG-I and RIG-I ligands. The literature proposes at least six different RIG-I ligands: (i) single strand with a 5'-triphosphate, (ii) double-stranded RNA with a 5'-triphosphate, (iii) 5' triphosphate single-stranded RNA with A- and U-rich 3'-sequences, (iv) double stranded RNA of intermediate length (>300 and <2000 bp) without 5'-triphosphate, (v) blunt-end short double-stranded RNA (23-30 bp) without 5'-triphosphate, and (vi) short double-stranded RNA (23-30 bp) with 5'-monophosphate. RIG-I thus seems promiscuous for a variety of different RNA molecules, very similar to the Toll like receptors, of which 10 family members are sufficient for the safe detection of the microbial cosmos. In the light of these outstanding publications, it seems an unlikely possibility that there is a fundamental shortcoming in the design of all studies. Looking closely, the only issue that comes to mind is the in vitro transcription technique used by all investigators without confirming the identity of RNA products. This technique, together with the different biological systems used, the lack of dose responses and of proper comparison of different published ligands and controls leave us with more questions than answers as to what the exact RIG-I ligand is, if in fact it exists. PMID- 19120478 TI - Viral sensors: diversity in pathogen recognition. AB - Innate sensors of viral infection detect viral products and initiate the signal cascades that lead to the antiviral response. Several proteins have been identified to play a role in this process, mostly members of the Toll-like receptor and retinoic acid-inducible gene I-like receptor families. These receptors have been demonstrated to function in part by recognizing a diverse yet unique repertoire of nucleic acid substrates. Upon recognition of their ligands, these sensors activate distinct signaling pathways that lead to the secretion of type I interferon and inflammatory cytokines. It remains to be seen, however, if these sensors are redundant or whether each serves a unique function. In this work, we review the current knowledge of viral sensors, speculate on how they may function in vivo, and explore the potential reasons for their diversity. PMID- 19120477 TI - Innate immunity to virus infection. AB - The innate immune system is essential for the initial detection of invading viruses and subsequent activation of adaptive immunity. Three classes of receptors, designated retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), Toll-like receptors (TLRs), and nucleotide oligomerization domain (NOD) like receptors (NLRs), sense viral components, such as double-stranded RNA (dsRNA), single-stranded RNA, and DNA. RLRs and TLRs play essential roles in the production of type I interferons (IFNs) and proinflammatory cytokines in cell type-specific manners. While the RLRs play essential roles in the recognition of RNA viruses in various cells, plasmacytoid dendritic cells utilize TLRs for detecting virus invasion. NLRs play a role in the production of mature interleukin-1 beta to dsRNA stimulation. Activation of innate immune cells is critical for mounting adaptive immune responses. In this review, we discuss recent advances in our understanding of the mechanisms of viral RNA recognition by these different types of receptors and its relation to acquired immune responses. PMID- 19120479 TI - Inflammasomes: guardians of cytosolic sanctity. AB - The innate immune system is critical in recognizing bacterial and viral infections to evoke a proper immune response. Certain members of the intracellular nucleotide-binding and oligomerization domain (NOD)-like receptor (NLR) family detect microbial components in the cytosol and trigger the assembly of large caspase-1-activating complexes termed inflammasomes. Autoproteolytic maturation of caspase-1 zymogens within these inflammasomes leads to maturation and secretion of the pro-inflammatory cytokines interleukin-1 beta (IL-1 beta) and IL-18. The NLR proteins ICE protease-activating factor (IPAF), NALP1b (NACHT domain-, leucine-rich repeat-, and PYD-containing protein 1b), and cryopyrin/NALP3 assemble caspase-1-activating inflammasomes in a stimulus dependent manner. Bacterial flagellin is sensed by IPAF, whereas mouse NALP1b detects anthrax lethal toxin. Cryopyrin/NALP3 mediates caspase-1 activation in response to a wide variety of microbial components and in response to crystalline substances such as the endogenous danger signal uric acid. Genetic variations in Nalp1 and cryopyrin/Nalp3 are associated with autoinflammatory disorders and increased susceptibility to microbial infection. Further understanding of inflammasomes and their role in innate immunity should provide new insights into the mechanisms of host defense and the pathogenesis of autoimmune diseases. PMID- 19120481 TI - Crosstalk between components of the innate immune system: promoting anti microbial defenses and avoiding immunopathologies. AB - Because it reaches full functional efficacy rapidly upon encounter with a pathogen, the innate immune system is considered as the first line of defense against infections. The sensing of microbes or of transformed or infected cells, through innate immune recognition receptors (referred to as activating I2R2), initiates pro-inflammatory responses and innate immune effector functions. Other I2R2 with inhibitory properties bind self-ligands constitutively expressed in host. However, this dichotomy in the recognition of foreign or induced self versus constitutive self by I2R2 is not always respected in certain non infectious conditions reminiscent of immunopathologies. In this review, we discuss that immune mechanisms have evolved to avoid inappropriate inflammatory disorders in individuals. Molecular crossregulation exists between components of I2R2 signaling pathways, and intricate interactions between cells from both innate and adaptive immune systems set the bases of controlled immune responses. We also pinpoint that, like T or B cells, some cells of the innate immune system must go through education processes to prevent autoreactivity. In addition, we illustrate how gene expression profiling of immune cell types is a useful tool to find functional homologies between cell subsets of different species and to speculate about unidentified functions of these cells in the responses to pathogen infections. PMID- 19120483 TI - Structure and regulation of cytoplasmic adapter proteins involved in innate immune signaling. AB - Initiation of the innate immune response requires agonist recognition by a pathogen recognition receptor. Following ligand binding, conformational rearrangement of the receptor creates a molecular scaffold from which signal transduction is propagated via complex cellular signaling pathways. This in turn leads to the induction of a pro-inflammatory immune response. A critical component of these signaling pathways is the homotypic interaction of receptor and adapter proteins via specific protein interaction domains. Within the innate immune signaling cascade, homotypic interactions between members of the death domain family and the Toll/interleukin-1 receptor domain are particularly important. Here we discuss the current understanding of the molecular basis of these homotypic receptor:adapter interactions and their role in innate immune signal transduction. PMID- 19120480 TI - Function of Nod-like receptors in microbial recognition and host defense. AB - Nucleotide oligomerization domain (NOD)-like receptors (NLRs) are a specialized group of intracellular proteins that play a critical role in the regulation of the host innate immune response. NLRs act as scaffolding proteins that assemble signaling platforms that trigger nuclear factor-kappaB and mitogen-activated protein kinase signaling pathways and control the activation of inflammatory caspases. Importantly, mutations in several members of the NLR family have been linked to a variety of inflammatory diseases consistent with these molecules playing an important role in host-pathogen interactions and the inflammatory response. In this review, we focus on the role of Nod1 and Nod2 in host defense and in particular discuss recent finding regarding the role of Nlrc4, Nlpr1, and Nlrp3 inflammasomes in caspase-1 activation and subsequent release of proinflammatory cytokines such as interleukin-1 beta. PMID- 19120482 TI - DAP10- and DAP12-associated receptors in innate immunity. AB - The DAP10 and DAP12 signaling subunits are highly conserved in evolution and associate with a large family of receptors in hematopoietic cells, including dendritic cells, plasmacytoid dendritic cells, neutrophils, basophils, eosinophils, mast cells, monocytes, macrophages, natural killer cells, and some B and T cells. Some receptors are able to associate with either DAP10 or DAP12, which contribute unique intracellular signaling functions. Studies of humans and mice deficient in these signaling subunits have provided surprising insights into the physiological functions of DAP10 and DAP12, demonstrating that they can either activate or inhibit immune responses. DAP10- and DAP12-associated receptors have been shown to recognize both host-encoded ligands and ligands encoded by microbial pathogens, indicating that they play an important role in innate immune responses. PMID- 19120484 TI - RNA-based viral immunity initiated by the Dicer family of host immune receptors. AB - Suppression of viral infection by RNA in a nucleotide sequence homology-dependent manner was first reported in plants in early 1990 s. Studies in the past 15 years have established a completely new RNA-based immune system against viruses that is mechanistically related to RNA silencing or RNA interference (RNAi). This viral immunity begins with recognition of viral double-stranded or structured RNA by the Dicer nuclease family of host immune receptors. In fungi, plants and invertebrates, the viral RNA trigger is processed into small interfering RNAs (siRNAs) to direct specific silencing of the homologous viral genomic and/or messenger RNAs by an RNaseH-like Argonaute protein. Deep sequencing of virus derived siRNAs indicates that the immunity against viruses with a positive-strand RNA genome is induced by Dicer recognition of dsRNA formed during the initiation of viral progeny (+)RNA synthesis. The RNA-based immune pathway in these organisms overlaps the canonical dsRNA-siRNA pathway of RNAi and may require amplification of viral siRNAs by host RNA-dependent RNA polymerase in plants and nematodes. Production of virus-derived small RNAs is undetectable in mammalian cells infected with RNA viruses. However, infection of mammals with several nucleus-replicating DNA viruses induces production of virus-derived microRNAs capable of silencing host and viral mRNAs as found for viral siRNAs. Remarkably, recent studies indicate that prokaryotes also produce virus-derived small RNAs known as CRISPR RNAs to guide antiviral defense in a manner that has yet to be defined. In this article, we review the recent progress on the identification and mechanism of the key components including viral sensors, viral triggers, effectors, and amplifiers, of the small RNA-directed viral immunity. We also highlight some of the many unresolved questions. PMID- 19120486 TI - Toll-like receptor signaling in the lysosomal pathways. AB - The lysosomal pathway digests material received by two main routes, phagocytosis and autophagy. Cells use phagocytosis to ingest extracellular particles by invaginations of the plasma membrane. In autophagy, a double membrane structure isolates portions of the cytoplasm to target it for degradation. During infection, phagocytes use both of these cellular functions to restrict microbial replication and at the same time to orchestrate an appropriate response against the invader. Toll-like receptor recognition of a pathogen initiates an innate immune response against the pathogen that includes production of inflammatory cytokines, upregulation of costimulatory molecules to prime an adaptive immune response, and activation of phagocytosis and autophagy. Signaling through this family of receptors also produces a hybrid response in which proteins that participate in autophagy are recruited to phagosomes, resulting in expedited microbial elimination. In this review, we discuss recent views on how Toll-like receptors direct microbes to final destruction by regulating the different pathways that lead to the lysosome. PMID- 19120485 TI - Autophagy and pattern recognition receptors in innate immunity. AB - Autophagy is a physiologically and immunologically controlled intracellular homeostatic pathway that sequesters and degrades cytoplasmic targets including macromolecular aggregates, cellular organelles such as mitochondria, and whole microbes or their products. Recent advances show that autophagy plays a role in innate immunity in several ways: (i) direct elimination of intracellular microbes by digestion in autolysosomes, (ii) delivery of cytosolic microbial products to pattern recognition receptors (PRRs) in a process referred to as topological inversion, and (iii) as an anti-microbial effector of Toll-like receptors and other PRR signaling. Autophagy eliminates pathogens in vitro and in vivo but, when aberrant due to mutations, contributes to human inflammatory disorders such as Crohn's disease. In this review, we examine these relationships and propose that autophagy is one of the most ancient innate immune defenses that has possibly evolved at the time of alpha-protobacteria-pre-eukaryote relationships, leading up to modern eukaryotic cell-mitochondrial symbiosis, and that during the metazoan evolution, additional layers of immunological regulation have been superimposed and integrated with this primordial innate immunity mechanism. PMID- 19120487 TI - Pattern recognition receptors and control of adaptive immunity. AB - The mammalian immune system effectively fights infection through the cooperation of two connected systems, innate and adaptive immunity. Germ-line encoded pattern recognition receptors (PRRs) of the innate immune system sense the presence of infection and activate innate immunity. Some PRRs also induce signals that lead to the activation of adaptive immunity. Adaptive immunity is controlled by PRR induced signals at multiple checkpoints dictating the initiation of a response, the type of response, the magnitude and duration of the response, and the production of long-term memory. PRRs thus instruct the adaptive immune system on when and how to best respond to a particular infection. In this review, we discuss the roles of various PRRs in control of adaptive immunity. PMID- 19120488 TI - Inflammatory signals in dendritic cell activation and the induction of adaptive immunity. AB - Pathogen invasion induces a rapid inflammatory response initiated through the recognition of pathogen-derived molecules by pattern recognition receptors (PRRs) expressed on both immune and non-immune cells. The initial wave of pro inflammatory cytokines and chemokines limits pathogen spread and recruits and activates immune cells to eradicate the invaders. Dendritic cells (DCs) are responsible for initiating a subsequent phase of immunity, dominated by the action of pathogen-specific T and B cells. As for the early pro-inflammatory response, DC activation is triggered by PRR signals. These signals convert resting DCs into potent antigen-presenting cells capable of promoting the expansion and effector differentiation of naive pathogen-specific T cells. However, it has been argued that signals from PRRs are not a prerequisite for DC activation and that pro-inflammatory cytokines have the same effect. Although this may appear like an efficient way to expand the number of DCs that initiate adaptive immunity, evidence is accumulating that DCs activated indirectly by inflammatory cytokines are unable to induce functional T-cell responses. Here, we review the differences between PRR-triggered and cytokine-induced DC activation and speculate on a potential role for DCs activated by inflammatory signals in tolerance induction rather than immunity. PMID- 19120489 TI - Microbe sensing, positive feedback loops, and the pathogenesis of inflammatory diseases. AB - The molecular apparatus that protects us against infection can also injure us by causing autoimmune or autoinflammatory disease. It now seems that at times, defects within the sensing arm of innate immunity contribute to diseases of this type. The initiation of an immune response is often microbe dependent and, in many cases, Toll-like receptor (TLR) dependent. Positive feedback loops triggering immune activation may occur when TLR signaling pathways stimulate host cells in an unchecked manner. Or, immune activation may persist because of failure to eradicate an inciting infection. Or on occasion, endogenous DNA may trigger specific immune responses that beget further responses in a TLR-dependent autoamplification loop. Specific biochemical defects that cause loop-related autoimmunity have been revealed by random germline mutagenesis and by gene targeting. We have also developed some insight into critical points at which feedback loops can be interrupted. PMID- 19120492 TI - Review series on helminths, immune modulation and the hygiene hypothesis: nematode coevolution with adaptive immunity, regulatory networks and the growth of inflammatory diseases. PMID- 19120490 TI - Systems biology of innate immunity. AB - Systems biology is the comprehensive and quantitative analysis of the interactions between all of the components of biological systems over time. Systems biology involves an iterative cycle, in which emerging biological problems drive the development of new technologies and computational tools. These technologies and tools then open new frontiers that revolutionize biology. Innate immunity is well suited for systems analysis, because the relevant cells can be isolated in various functional states and their interactions can be reconstituted in a biologically meaningful manner. Application of the tools of systems biology to the innate immune system will enable comprehensive analysis of the complex interactions that maintain the difficult balance between host defense and inflammatory disease. In this review, we discuss innate immunity in the context of the systems biology concepts, emergence, robustness, and modularity, and we describe emerging technologies we are applying in our systems-level analyses. These technologies include genomics, proteomics, computational analysis, forward genetics screens, and analyses that link human genetic polymorphisms to disease resistance. PMID- 19120493 TI - Review series on helminths, immune modulation and the hygiene hypothesis: the broader implications of the hygiene hypothesis. AB - Man has moved rapidly from the hunter-gatherer environment to the living conditions of the rich industrialized countries. The hygiene hypothesis suggests that the resulting changed and reduced pattern of exposure to microorganisms has led to disordered regulation of the immune system, and hence to increases in certain inflammatory disorders. The concept began with the allergic disorders, but there are now good reasons for extending it to autoimmunity, inflammatory bowel disease, neuroinflammatory disorders, atherosclerosis, depression associated with raised inflammatory cytokines, and some cancers. This review discusses these possibilities in the context of Darwinian medicine, which uses knowledge of evolution to cast light on human diseases. The Darwinian approach enables one to correctly identify some of the organisms that are important for the 'Hygiene' or 'Old Friends' hypothesis, and to point to the potential exploitation of these organisms or their components in novel types of prophylaxis with applications in several branches of medicine. PMID- 19120494 TI - Review series on helminths, immune modulation and the hygiene hypothesis: how might infection modulate the onset of type 1 diabetes? AB - The development of type 1 diabetes is influenced by both genetic and environmental factors. The current rise in the incidence of diabetes is occurring more rapidly than can be accounted for by genetic change, highlighting the influence of environmental modifiers. Considerable effort has been expended to identify infectious agents that might be responsible for this rise in incidence, but no single infectious agent has been linked to this dramatic increase in type 1 diabetes. There has been increasing interest in the possibility that infections of historical importance that might have shaped our immune systems over evolutionary time may also have played a role in down-modulating some autoimmune and allergic disorders. In this review, some of the ways in which certain organisms might have influenced the onset of autoimmunity are discussed. PMID- 19120497 TI - After thirty years of Indoor Air conferences, from 1978 Copenhagen to 2008 Copenhagen, where do we stand? Editorial. PMID- 19120495 TI - Review series on helminths, immune modulation and the hygiene hypothesis: immunity against helminths and immunological phenomena in modern human populations: coevolutionary legacies? AB - Although the molecules and cells involved in triggering immune responses against parasitic worms (helminths) remain enigmatic, research has continued to implicate expansions of T-helper type 2 (Th2) cells and regulatory T-helper (T(reg)) cells as a characteristic response to these organisms. An intimate association has also emerged between Th2 responses and wound-healing functions. As helminth infections in humans are associated with a strong Th2/T(reg) immunoregulatory footprint (often termed a 'modified Th2' response), plausible links have been made to increased susceptibility to microbial pathogens in helminth-infected populations in the tropics and to the breakdowns in immunological control (allergy and autoimmunity) that are increasing in frequency in helminth-free developed countries. Removal of helminths and their anti-inflammatory influence may also have hazards for populations exposed to infectious agents, such as malaria and influenza, whose worst effects are mediated by excessive inflammatory reactions. The patterns seen in the control of helminth immunity are discussed from an evolutionary perspective. Whilst an inability to correctly regulate the immune system in the absence of helminth infection might seem highly counter-adaptive, the very ancient and pervasive relationship between vertebrates and helminths supports a view that immunological control networks have been selected to function within the context of a modified Th2 environment. The absence of immunoregulatory stimuli from helminths may therefore uncover maladaptations that were not previously exposed to selection. PMID- 19120496 TI - Review series on helminths, immune modulation and the hygiene hypothesis: mechanisms underlying helminth modulation of dendritic cell function. AB - Dendritic cells (DCs) play a central role in activating CD4 T (T helper, Th) cells. As a component of their response to pathogen-associated stimuli, DCs produce cytokines and express surface molecules that provide important cues to modulate the effector functions of responding Th cells. Much is known of how DCs respond to, and influence immune response outcome to, bacterial and viral pathogens. However, relatively little is understood about how DCs respond to helminth parasites. This is an area of considerable interest since it impacts our understanding of the initiation of Th2 responses, which are stereotypically associated with helminth infections, and the regulation of allergic and autoimmune pathologies which evidence suggests are less severe or absent in individuals infected with helminths. This review attempts to summarize our understanding of the effects of helminth products on dendritic cell biology. PMID- 19120498 TI - Indoor airborne endotoxin assessment in homes of Paris newborn babies. AB - The aims of this study were first to assess airborne endotoxin levels in the dwellings of 162 newborns living in Paris twice during a 1-year period, and second, to identify predictors for endotoxin concentrations using questionnaire data in relation to housing factors and living conditions. Air samples were collected on a glass fiber filter in polystyrene filter holders, using a pump at a flow rate of 3.5 l/min for 24 h placed in the main room of the home. Endotoxin levels were measured using a chromogenic kinetic Limulus Amoebocyte Lysate test. Geometric means (geometric standard deviation) of airborne endotoxin levels at two different visits were respectively 0.509 (4.289) EU/m3 and 0.557 (3.029) EU/m3. Airborne endotoxin levels were significantly increased: (i) in cold season (P = 0.024), with (ii) the presence of visible cockroaches in the previous 12 months at home (P < 0.001), (iii) increased number of inhabitants per square meter (P = 0.012), (iv) the high frequency of cleaning with the floor cloths (P = 0.0014), and (v) the low frequency of vacuuming (P = 0.0045). This study provided for the first time airborne endotoxin levels issued from repeated measurements in Paris dwellings. PRACTICAL IMPLICATIONS This analysis contributed to identify a few factors that determined indoor airborne endotoxin levels. However, the predictive model including housing factors and living conditions poorly estimated endotoxin levels. Consequently, multiple samples and longer sampling periods might improve the estimate of long-term airborne endotoxin exposure especially its variability, in cohort studies. PMID- 19120499 TI - Indoor air quality assessment in and around urban slums of Delhi city, India. AB - The present study aims at investigating the indoor air quality (IAQ) in selected households in one of the urban slums i.e. the Nizamuddin slums in Delhi, the capital city of India. The study includes investigations and assessments on associated health effects on the occupants living in inefficiently designed houses having poor ventilation. The monitoring of indoor air pollutants e.g. the respirable suspended particulate matter (RSPM), the carbon dioxide (CO2), the carbon monoxide (CO), the sulphur dioxide (SO2) and the nitrogen dioxide (NO2) for all three seasons i.e. summer (April-June 2004), rainy (July-September 2004) and winter (December 2004-February 2005) have been conducted. In addition, the spirometry tests on the occupants, particularly the womenfolk and children have been performed to determine the incidence of acute respiratory infections (ARI). Questionnaire survey has also been conducted in the households during the study period to investigate the sick building syndrome (SBS). The study reveals maximum concentration of indoor air pollutants in households during winters (December 2004-February 2005) associated with aggravated respiratory problems like cough, phlegm, wheezing, and breathlessness among occupants particularly the women occupants. Besides, decrement in lung function indices (i.e. FVC and/or FEV1) due to increased concentrations of RSPM and CO2 indoors during winter period has also been observed in the women respondents. The study concludes that women and children indoors are most vulnerable to respiratory problems compared to other sexes. A high SBS score is observed in these 'urban poor' households because of inadequate ventilation. PRACTICAL IMPLICATIONS: 'High indoor airborne pollutants during winter are associated with respiratory problems for women and children in houses in urban slum in Delhi. The work demonstrated the need of further studies of indoor air quality for the 'urban poor' in developing countries.' PMID- 19120500 TI - Particle dose estimation from frying in residential settings. AB - Fumes produced during frying have been implicated as a potential cause for the increased incidence of adenocarcinoma. Particulate matter exposure has also been linked with other pulmonary and coronary disease. This study investigated the contribution of frying in residential settings to ultrafine and fine particulate matter (UFP, PM2.5, respectively) exposure in homes. Production rates of 44 +/- 26 particles (pt)/cm3 s (mean +/- standard deviation) and 0.13 +/- 0.12 microg/m3 s were found for UFP and PM2.5, respectively, from frying a variety of foods at medium heat in a loft-style apartment. Rates of 290 +/- 150 pt/cm3 s and 3.5 +/- 4.9 microg/m3 s were found for UFP and PM2.5, respectively, from frying with vegetable oil alone in five homes; the higher rates were ascribed to differences between the homes rather than the absence of food. The elimination of UFP and PM2.5 was found to be primarily through exhaust fans in these homes, and it was found to follow a first-order process with an elimination rate constant of 6.1 x 10(-4) +/- 2.5 x 10(-4) s(-1). The dose to an individual from frying was estimated based on the measured production and elimination rates and found to be significant when compared with the typical daily dose incurred within a home because of outside sources. PRACTICAL IMPLICATIONS: The contribution of indoor sources to particulate matter exposure in homes remains poorly understood. Yet common household activities such as frying may produce substantial concentrations of potentially toxic particles. Because of the potential adverse health impacts associated with exposure to air pollution, potentially vulnerable individuals may be advised to remain indoors at certain times so as to reduce their overall exposure. Such interventions can be negated without proper guidance regarding the exposure involved in various indoor activities such as cooking. This paper outlines a methodology to estimate the dose to particulate matter incurred during frying and shows that this can represent a significant source of daily exposure. PMID- 19120501 TI - Effect of floor surface temperature on blood flow and skin temperature in the foot. AB - A total of 16 healthy college students participated as subjects to elucidate the hypothesis that blood flow and skin temperature in foot are affected by the floor surface temperature. The floor surface temperature was controlled by varying the temperature of water (tw) flowing underneath the floor, and it ranged from tw 15 to 40 degrees C at 5 degrees C intervals. The blood flow rate was measured in the dorsal right toe, and skin temperatures were measured for 60 min at 8 points: the neck, right scapular, left hand, right shin, left bottom of the toe, right instep, left finger, and rectum. The blood flow rate in the foot tissue was increased until the foot skin temperature warmed up to 34 degrees C (P = 0.000). The final skin temperatures on the bottom of the toe were 19.4 +/- 2.44 degrees C for tw 15 degrees C, 22.4 +/- 2.45 degrees C for tw 20 degrees C, 24.8 +/- 2.80 degrees C for tw 25 degrees C, 27.7 +/- 2.13 degrees C for tw 30 degrees C, 30.6 +/- 2.06 degrees C for tw 35 degrees C, 33.2 +/- 1.45 degrees C for tw 40 degrees C, 34.2 +/- 1.55 degrees C for tw 45 degrees C, and 35.2 +/- 1.65 degrees C for tw 50 degrees C. Considering blood flow and comfort, the partial floor heating system is suggested and the recommended floor surface temperature range is 27-33 degrees C. PRACTICAL IMPLICATIONS: A warm floor surface can serve to satisfy occupants when the ambient temperature maintained at 20 degrees C which represents an energy conscious temperature. A warm floor can induce high blood perfusion in the feet and consequently improve an occupant's health by treating many vascular-related disorders. Even in a well-insulated residential building, a partially heated floor system could prevent overheating while providing surface warmth. PMID- 19120504 TI - Themed issue: pulp biology and clinical practice. PMID- 19120502 TI - Sources of nitrogen dioxide (NO2) in New Zealand homes: findings from a community randomized controlled trial of heater substitutions. AB - Houses in New Zealand have inadequate space heating and a third of households use unflued gas heaters. As part of a large community intervention trial to improve space heating, we replaced ineffective heaters with more effective, non-polluting heaters. This paper assesses the contribution of heating and household factors to indoor NO2 in almost 350 homes and reports on the reduction in NO2 levels due to heater replacement. Homes using unflued gas heaters had more than three times the level of NO2 in living rooms [geometric mean ratio (GMR) = 3.35, 95% CI: 2.83 3.96, P < 0.001] than homes without unflued gas heaters, whereas homes using gas stove-tops had significantly elevated living room NO2 levels (GMR = 1.42, 95% CI: 1.05-1.93, P = 0.02). Homes with heat pumps, flued gas heating, or enclosed wood burners had significantly lower levels of NO2 in living areas and bedrooms. In homes that used unflued gas heaters as their main form of heating at baseline, the intervention was associated with a two-third (67%) reduction in NO2 levels in living rooms, when compared with homes that continued to use unflued gas heaters. Reducing the use of unflued gas heating would substantially lower NO2 exposure in New Zealand homes. PRACTICAL IMPLICATIONS: Understanding the factors influencing indoor NO2 levels is critical for the assessment and control of indoor air pollution. This study found that homes that used unflued gas combustion appliances for heating and cooking had higher NO2 levels compared with homes where other fuels were used. These findings require institutional incentives to increase the use of more effective, less polluting fuels, particularly in the home environment. PMID- 19120505 TI - Assessment of pulp vitality: a review. AB - BACKGROUND: One of the greatest diagnostic challenges in clinical practice is the accurate assessment of pulp status. This may be further complicated in paediatric dentistry where the practitioner is faced with a developing dentition, traumatized teeth, or young children who have a limited ability to recall a pain history for the tooth in question. A variety of pulp testing approaches exist, and there may be confusion as to their validity or appropriateness in different clinical situations. AIM: The aim of this paper is to provide the clinician with a comprehensive review of current pulp testing methods. A key objective is to highlight the difference between sensitivity testing and vitality testing. A biological basis for pulp testing is also provided to allow greater insight into the interpretation of pulp testing results. The rationale for, and methods of, assessing pulpal blood flow are described. PMID- 19120506 TI - Pulpal status of human primary teeth with physiological root resorption. AB - OBJECTIVE: The overall aim of this study was to determine whether any changes occur in the pulpal structure of human primary teeth in association with physiological root resorption. METHODS: The experimental material comprised 64 sound primary molars, obtained from children requiring routine dental extractions under general anaesthesia. Pulp sections were processed for indirect immunofluorescence using combinations of: (i) protein gene product 9.5 (a general neuronal marker); (ii) leucocyte common antigen CD45 (a general immune cell marker); and (iii) Ulex europaeus I lectin (a marker of vascular endothelium). Image analysis was then used to determine the percentage area of staining for each label within both the pulp horn and mid-coronal region. Following measurement of the greatest degree of root resorption in each sample, teeth were subdivided into three groups: those with physiological resorption involving less than one-third, one-third to two-thirds, and more than two-thirds of their root length. RESULTS: Wide variation was evident between different tooth samples with some resorbed teeth showing marked changes in pulpal histology. Decreased innervation density, increased immune cell accumulation, and increased vascularity were evident in some teeth with advanced root resorption. Analysis of pooled data, however, did not reveal any significant differences in mean percentage area of staining for any of these variables according to the three root resorption subgroups (P > 0.05, analysis of variance on transformed data). CONCLUSIONS: This investigation has revealed some changes in pulpal status of human primary teeth with physiological root resorption. These were not, however, as profound as one may have anticipated. It is therefore speculated that teeth could retain the potential for sensation, healing, and repair until advanced stages of root resorption. PMID- 19120507 TI - Histological comparison of pulpal inflammation in primary teeth with occlusal or proximal caries. AB - OBJECTIVE: A number of clinical and histological studies have investigated caries related changes in the primary tooth pulp, but the effect of caries site, as a clinical variable, has not been previously considered. This study sought to compare inflammatory changes within the pulp of primary molars according to the location of the caries lesion (occlusal or proximal). METHODS: Eighty-three primary molars were extracted under general anaesthesia for caries and/or orthodontic reasons, and were split immediately after removal and fixed in 10% formalin. Teeth were then decalcified, sectioned, and stained with haematoxylin and eosin for histological examination using light microscopy. Caries depth was measured using a graticle, and the site of the caries lesion noted as occlusal or proximal. Samples were further classified into one of five subgroups according to the observed degree of pulpal inflammation. RESULTS: Key findings were that where caries depth was less than 50% of the total dentine thickness, there were no significant differences in inflammatory status according to caries site. In contrast, marked inflammatory changes were significantly more likely throughout the coronal pulp of teeth with proximal caries compared to teeth with occlusal caries where caries depth was equal to, or greater than, 50% of the total dentine thickness (P = 0.017, Fisher's exact test). CONCLUSION: Primary teeth with proximal carious lesions extending more than 50% through the dentine thickness appear to have more extensive inflammatory pulpal changes than teeth with occlusal caries of a similar depth. This finding has clinical implications and may help inform treatment decisions in the management of primary teeth with deep carious lesions. PMID- 19120508 TI - Mineral trioxide aggregate in paediatric dentistry. AB - OBJECTIVE: The aim of this study was to present a review of the reported literature on: (i) the physical and chemical properties; and (ii) clinical applications of mineral trioxide aggregate (MTA) in the practice of paediatric dentistry. METHOD: Electronic literature search of scientific papers from January 1993 to June 2008 was carried out on the MEDLINE, Embase, Entrez Pubmed, and Scopus databases using specific key words. The search yielded 448 papers, out of which 100 were identified as conforming to the applied criteria. These papers formed the basis of the review and the clinical scenarios presented which demonstrate the application of MTA in the practice of paediatric dentistry. CONCLUSION: Paediatric dentists have successfully employed MTA in a variety of endodontic/restorative applications since the late 1990s. Clinical impressions have generally been favourable and support the findings of laboratory and animal based investigations. Very few clinical studies have been reported so far in humans, and although these have been positive, the body of research is currently insufficient to enable a meaningful systematic review and meta-analysis. PMID- 19120509 TI - Dental pulp stem cells: what, where, how? AB - INTRODUCTION: It is now accepted that progenitor/stem cells reside within the post-natal dental pulp. Studies have identified several niches of multipotent mesenchymal progenitor cells, known as dental pulp stem cells, which have a high proliferative potential for self-renewal. These progenitor stem cells are now recognized as being vital to the dentine regeneration process following injury. Understanding the nature of these progenitor/stem cell populations in the pulp is important in determining their potentialities and development of isolation or recruitment strategies for use in regeneration and tissue engineering. Characterization of these cells, and determination of their potentialities in terms of specificity of regenerative response, may help direct new clinical treatment modalities. Such novel treatments may involve controlled direct recruitment of the cells in situ and possible seeding of stem cells at sites of injury for regeneration or use of the stem cells with appropriate scaffolds for tissue engineering solutions. Such approaches may provide an innovative and novel biologically based new generation of clinical materials and/or treatments for dental disease. AIM: This study aimed to review the body of knowledge relating to stem cells and to consider the possibility of these cell populations, and related technology, in future clinical applications. PMID- 19120510 TI - Systemic inflammatory response syndrome after percutaneous nephrolithotomy: an assessment of risk factors. AB - OBJECTIVES: To analyze the risk factors for systemic inflammatory response syndrome (SIRS) after percutaneous nephrolithotomy (PCNL) and to quantitatively predict the probability of SIRS after PCNL. METHODS: Medical records on 209 patients who underwent PCNL were retrospectively analyzed. The chi2 test, the t test and a logistic regression model were used to identify key risk factors of SIRS after PCNL. A predictive equation was then formulated to assess the risk of SIRS according to the results from the logistic model. Subsequently, the accuracy of the equation by calculating sensitivity, specificity, overall correct percentage, and positive and negative predictive values was tested. RESULTS: The incidence of SIRS after PCNL was 23.4%. The key risk factors for SIRS following PCNL were: the number of tracts, receipt of a blood transfusion, stone size, and presence of pyelocaliectasis. Other factors added no independent risk to the development of SIRS. The calculated values for sensitivity, specificity, overall percentage correct, positive predictive value and negative predictive value were 44.9%, 95.0%, 83.3%, 73.3%, and 84.9%, respectively. CONCLUSIONS: Number of tracts, receipt of a blood transfusion, stone size and presence of pyelocaliectasis are identified as the key risk factors for SIRS after PCNL. The predictive equation allows for an individualized and quantitative assessment of the probability of SIRS after PCNL. PMID- 19120511 TI - Genetic polymorphism of sulfotransferase 1A1, cigarette smoking, hazardous chemical exposure and urothelial cancer risk in a Taiwanese population. AB - OBJECTIVES: To investigate the association between genetic polymorphism of sulfotransferase1A1 (SULT1A1), cigarette smoking, hazardous chemical exposure and urothelial cancer risk in a Taiwanese population. METHODS: In a hospital-based case-control study, a total of 300 urothelial cancer (UC) cases and 300 cancer free controls frequency-matched by age and gender were recruited from September 1998 to December 2005. The SULT1A1 arginine213histidine (Arg213His) polymorphism was genotyped using a polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: We found that the significantly increased UC risks of ever smokers and heavy smokers (> or =28 pack-years) were 2.1 (95% confidence interval [CI] = 1.4-3.3) and 2.2 (95% CI = 1.3-3.6), respectively. An increased UC risk of 1.8 (95% CI = 0.8-3.8) was observed among individuals with more than one item of hazardous chemical exposure, but it was not statistically significant. Compared with study subjects carrying the SULT1A1 Arg/Arg genotype, those with SULT1A1 Arg/His or His/His genotypes have a significantly decreased UC risk (Odds ratio [OR] = 0.5, 95% CI = 0.3-0.8). Heavy smokers carrying the SULT1A1 Arg/Arg genotype have a significantly increased UC risk (OR = 5.2, 95% CI = 2.3-11.6). Individuals who had been exposed to more than one item of hazardous chemicals and who carried the SULT1A1 Arg/Arg genotype have a significantly increased UC risk (OR = 3.7, 95% CI = 1.4-9.7). The highest significant increased UC risk (OR = 16.1, 95% CI = 2.9-87.2) was observed among ever smokers with hazardous chemical exposure and the SULT1A1 Arg/Arg genotype. CONCLUSIONS: SULT1A1 Arg213His polymorphism is associated with the development of UC, especially among cigarette smokers exposed to hazardous chemicals. PMID- 19120512 TI - Hand-assisted laparoscopic radical cystectomy and extracorporeal urinary diversion: experience with 31 cases. AB - OBJECTIVES: To report our techniques and experience with hand-assisted laparoscopic radical cystectomy and extracorporeal urinary diversion for bladder cancer. METHODS: Between May 2004 and November 2007, 31 patients (mean age 61.3 years, range 40-79) underwent hand-assisted laparoscopic radical cystectomy with extracorporeal urinary diversion for bladder cancer. Five patients had previously undergone abdominal surgeries. Data were collected with respect to patient demographics, perioperative outcomes and short-term oncological follow up. RESULTS: Twenty-four patients underwent an ileal conduit and seven patients underwent an orthotopic neobladder. Mean operative time was 365.7 min (range 245 to 530). Estimated blood loss was 250.9 cc (range 100 to 500), with a transfusion rate of 9.7%. Oral liquids were resumed at 4.3 days and the mean hospital stay was 19.7 days. There were no intraoperative complications. Postoperative early complications (within 30 days of surgery) occurred in six patients (19.4%). Two wound infections, one urinary leak, one wound dehiscence, one bowel obstruction and one alimentary tract hemorrhage were all treated conservatively. Late complications occurred in three patients (two parastomal hernias and one ureteroenteric stricture). With a mean follow up of 18 months, 27 patients had no evidence of disease. One patient died because of cancer and one died for unrelated causes. One was alive with local recurrences and one with lung metastasis. CONCLUSIONS: Hand-assisted laparoscopic radical cystectomy is a safe, reproducible and minimally invasive option for bladder cancer patients. PMID- 19120513 TI - Naftopidil versus tamsulosin hydrochloride for lower urinary tract symptoms associated with benign prostatic hyperplasia with special reference to the storage symptom: a prospective randomized controlled study. AB - OBJECTIVES: In order to compare the clinical efficacy of naftopidil (Naf) and tamsulosin hydrochloride (Tam), which differ in their selectivity to alpha receptor subtypes, we performed a multi-center prospective randomized controlled study. METHODS: Men complaining of lower urinary tract symptoms due to benign prostatic hypertrophy, were randomized into two treatment groups: one receiving 50 mg Naftopidil daily (Naf group, n = 31 pts), and one receiving 0.2 mg Tam once daily (Tam group, n = 28 pts). Baseline symptom scores were compared to those at 2 weeks and at the end of the observation period (6-8 weeks). RESULTS: In the Naf group at 2 weeks, the score of the daytime frequency significantly improved from 3.5 to 2.2 (P = 0.03), and the score of nocturia improved significantly from 3.5 to 2.2 (P = 0.0004), respectively. In the Tam group at 2 weeks, however, no significant improvement was noted in the increased score of daytime frequency (P = 0.1) or nocturia (P = 0.2). At 2 weeks, the storage symptom score of the frequency to the combined score of daytime frequencies and the score of nocturia was better in the Naf group (improved from 7.0 to 4.4, P = 0.0017) than in the Tam group (from 6.8 to 4.9, P = 0.08) (P < 0.05). At 6-8 weeks, the effects of the two drugs on lower urinary tract symptoms were comparable. CONCLUSIONS: Naf demonstrated a significant early response to improve storage symptoms at 2 weeks, including daytime frequency and nocturia, compared with Tam. PMID- 19120514 TI - Three-dimensional 26-core biopsy-based patient selection criteria for nerve sparing radical prostatectomy. AB - OBJECTIVES: Most of the previously reported patient selection criteria for nerve sparing radical prostatectomy were based on conventional sextant biopsy, and those based on extended biopsy have been scarcely investigated. In the current study, we developed patient selection criteria for nerve-sparing RP based on the three-dimensional 26-core (3D26) biopsy-derived variables. METHODS: We evaluated 202 non-nerve-spared sides in 109 consecutive patients in whom prostate cancer was diagnosed by the 3D26 biopsy and who underwent RP without neoadjuvant treatment. Associations of clinical and pathological variables with side-specific posterolateral extraprostatic extension (SS-PL-EPE) were analyzed. Subgroup analysis using transperineal 14-core (TP14) and transrectal 12-core (TR12) biopsies as representative subsets of the 3D26 biopsy was also performed. RESULTS: Maximum cancer length in positive cores > or =5 mm and biopsy Gleason score > or =4 + 3 were independent and were significant risk factors of SS-PL-EPE in the 3D26 cohort at multivariate analysis. In the prostatic side with none, one, and two risk factors, the incidences of SS-PL-EPE were 0, 14 and 52% in the 3D26 cohort, 3.4,15 and 57% in the TP14 cohort and 2.6, 20 and 61% in the TR12 cohort, respectively. CONCLUSIONS: We developed simple patient selection criteria for nerve-sparing RP. According to our criteria, the nerve-sparing side can be selected in the majority of patients who undergo the 3D26, TP14 or TR12 biopsy with a less-than-4% risk of SS-PL-EPE. PMID- 19120515 TI - Effects of a novel beta(3)-adrenoceptor agonist, AJ-9677, on relaxation of the detrusor muscle: an in vitro study. AB - OBJECTIVES: To examine the relaxant effects of AJ-9677, a novel beta(3) adrenoceptor agonist, on the isolated rat, monkey and human detrusor muscle. METHODS: The isolated detrusor strips of rats, monkeys and humans were mounted in organ baths containing Krebs solution. By the cumulative addition of beta adrenoceptor agonists (isoproterenol, AJ-9677, CL 316,243 and salbutamol in rats; isoproterenol, AJ-9677 and CL 316,243 in monkeys and humans), concentration relaxation curves were obtained. The maximal relaxation responses and pEC(50) values were calculated. In rats, concentration-relaxation curves to isoproterenol and AJ-9677 were obtained in the presence and absence of propranolol or SR 59230A. RESULTS: Isoproterenol, AJ-9677, CL 316,243 and salbutamol induced concentration-dependent relaxation in rats. The rank order of their relaxing potency in the rat detrusor muscle was AJ-9677 > isoproterenol > CL 316,243 > salbutamol. Isoproterenol and AJ-9677 also produced a concentration-dependent relaxation with high potency in monkeys and humans, whilst CL 316,243 had low relaxing potency. According to the antagonist studies in rats, propranolol and SR 59230A caused a rightward shift of the concentration-relaxation curves to isoproterenol or AJ-9677, respectively. CONCLUSIONS: AJ-9677 has a high relaxant potency on the rat, monkey and human detrusor smooth muscle, and it may have the potential to treat overactive bladder. PMID- 19120516 TI - Renal cell carcinoma with a huge solitary metastasis to the contralateral adrenal gland: a case report. AB - Renal cell carcinoma (RCC) is capable of metastasizing to several organs. Synchronous isolated contralateral adrenal metastasis of the primary RCC is, however, very rare. Herein we report a case of RCC with a huge solitary metastasis to the contralateral adrenal gland that was surgically treated. We scheduled nephrectomy for the left primary RCC and adrenalectomy for the right adrenal tumor. However, at surgery we found a huge right adrenal tumor that had invaded the right kidney, right renal vein, and inferior vena cava. Therefore right nephrectomy was performed simultaneously with resection and reconstruction of the inferior vena cava. Pathological findings demonstrated that the left renal tumor and right adrenal tumor had the same histology. Although the patient required hemodialysis, he remains well at six months postoperatively. So far, there have been only two cases of a solitary contralateral metastatic adrenal tumor that was larger than the primary RCC, thus the present case is the third one. PMID- 19120517 TI - Primary large cell neuroendocrine carcinoma of the urinary bladder. AB - Large cell neuroendocrine carcinoma (LCNEC) of the urinary bladder is rare. It is a type of neuroendocrine carcinoma morphologically distinct from small cell carcinoma. We report here a case of primary LCNEC of the urinary bladder. We observed a very large invasive tumor, which was not able to be detected three months previously by cystoscopy or computed tomography. The tumor cells morphologically and immunohistochemically resembled that of pulmonary LCNEC. With prompt cystoprostatectomy and chemotherapy, the patient is free of disease 16 months after diagnosis. Although LCNEC is usually very aggressive, it may be controlled by early diagnosis and treatment. PMID- 19120518 TI - Varicocele complicating spontaneous arteriovenous fistula. AB - We describe herein a rare case of a varicocele complicating spontaneous arteriovenous fistula. A 40-year-old man was referred to our hospital in November 2006, complaining of a non-tender mass in the left scrotum at the age of 15 and thereafter. On examination, his left scrotum revealed a large varicocele, but no manifest superficial thrill was noted. Scrotal ultrasonograpy revealed approximately 7 cm large varicocele. Computed tomography angiography revealed the existence of an arteriovenous fistula between the left testicular artery and the veins of the left pampiniform plexus. We laparoscopically carried out internal spermatic vessels ligation under the diagnosis of a varicocele complicating a spontaneous arteriovenous fistula. The postoperative course was uneventful. At 18 months postoperatively, the varicocele and fistula had not recurred. PMID- 19120520 TI - New years greetings. PMID- 19120521 TI - How to write titles and abstracts for readers. PMID- 19120522 TI - Japanese guideline for diagnosis and treatment of interstitial cystitis. AB - Interstitial cystitis (IC) is a disease of the urinary bladder with lower urinary tract symptoms such as urinary frequency, bladder hypersensitivity and/or bladder pain and resultant serious impairment of quality of life. In Japan, assuming that IC is very rare, research activity and medical care of IC have been sparse until 2001, when the Society of Interstitial Cystitis of Japan (SICJ) and a patient support group were founded.(1,2) Subsequently the International Consultation on Interstitial Cystitis Japan (ICICJ) was held in Kyoto in 2003.(3) On the other hand, the etiology of IC has not been well clarified, which complicates its diagnosis and treatment at clinical settings. We have thus developed the Japanese Clinical Guideline,(4) which is targeted at healthcare professionals including specialists in urology and women's health care who may engage in the diagnosis and treatment of IC. This article is the English translation of a shortened version of the Guideline for convenience of readers worldwide. PMID- 19120523 TI - Bladder cancer biomarkers and their role in surveillance and screening. AB - Early detection of bladder cancer and its recurrences is essential for improved prognosis and long-term survival. The detection and follow-up of these patients is currently based on cystoscopy, which is expensive and invasive, and, in most cases, cytology, which is non-invasive but not very sensitive. During recent years, many urine-based tests have been developed and tested in different patient populations. In this review we discuss new developments for biomarkers in bladder cancer that have potential use in surveillance and screening. In almost all publications authors compare sensitivity of the test with a concomitantly executed cystoscopy, for example, determine cross-sectional sensitivity. However, it has also been shown that false positive test results may be followed by a positive cystoscopy in the near future, showing that cystoscopy itself does not provide 100% sensitivity. This suggests that for a proper evaluation of urine based tests, longitudinal studies should be carried out and the results communicated to the urologist. PMID- 19120525 TI - Estrogen and urinary incontinence. AB - Hormone therapy (HT) has been one of the most commonly used methods to treat postmenopausal disorders including lower urinary tract symptoms such as urinary incontinence (UI). Although it has been suggested that HT improves urinary tract symptoms, little evidence has so far been presented to support this. Recently however, large randomized clinical trials have shown the adverse effects of estrogen on UI and indicate that HT should not be used for prevention or relief of UI in postmenopausal women. Here we review the effects of HT on UI in the light of these clinical trials. PMID- 19120524 TI - Neuroendocrine differentiation in the progression of prostate cancer. AB - Neuroendocrine (NE) cells originally exist in the normal prostate acini and duct, regulating prostatic growth, differentiation and secretion. Clusters of malignant NE cells are found in most prostate cancer (PCa) cases. NE differentiation (NED) is the basic character of the prostate, either benign or malignant. NE cells hold certain peptide hormones or pro-hormones, which affect the target cells by endocrine, paracrine, autocrine and neuroendocrine transmission in an androgen independent fashion due to the lack of androgen receptor. NED is accessed by immunohistochemical staining or measurement of serum levels of NE markers. The extent of NED is associated with progression and prognosis of PCa. Chromogranin A (CGA) is the most important NE marker. In metastatic PCa, pretreatment serum CGA levels can be a predictor for progression and survival after endocrine therapy. It is recommended to measure longitudinal change in serum CGA. The NE pathway can also be a therapeutic target. PMID- 19120526 TI - Neural tube defects: prevalence, etiology and prevention. AB - Spina bifida and anencephaly, the common form of neural tube defects, affect approximately 300 000 newborns each year worldwide. The effectiveness of folic acid supplementation in preventing their occurrence or recurrence has been unambiguous since 1991. In general, the prevalence of these abnormalities has decreased in the past 20 to 30 years because of periconceptional folate supplementation, food fortification in several countries, avoiding exposure to environmental factors, and increased accuracy of prenatal screening for fetal anomalies. Women who are planning to conceive should be informed about the importance of folic acid in fetal development and advised to take 400 microg/day of folic acid supplements. Food fortification with folic acid will ultimately be necessary to reduce the number of patients. Primary prevention of neural tube defects by the periconceptional intake of folic acid is a major public health opportunity and has wide implications in reducing the mortality and morbidity of offspring. PMID- 19120527 TI - Computed tomography angiogram: accuracy in renal surgery. AB - OBJECTIVES: To determine the sensitivity and specificity of computed tomography angiogram (CTA) in detecting number and location of renal arteries and veins as well as crossing vessels causing uretero-pelvic junction obstruction (UPJO), and to determine if this can be used in decision-making algorithms for treatment of UPJO. METHODS: A prospective study was carried out in patients undergoing open, laparoscopic and robotic renal surgery from April 2005 until October 2006. All patients were imaged using CTA with 1.25 collimation of arterial and venous phases. Each multi-detector CTA was then read by one radiologist and his results were compared prospectively with the actual intra-operative findings. RESULTS: Overall, 118 patients were included. CTA had 93% sensitivity, 77% specificity and 90% overall accuracy for detecting a single renal artery, and 76% sensitivity, 92% specificity and 90% overall accuracy for detecting two or more renal arteries (Pearson chi(2) = 0.001). There was 95% sensitivity, 84% specificity and 85% overall accuracy for detecting the number of renal veins. CTA had 100% overall accuracy in detecting early dividing renal artery (defined as less than 1.5 cm branching from origin), and 83.3% sensitivity, specificity and overall accuracy in detecting crossing vessels at UPJ. The percentage of surgeons stating CTA to be helpful as pre-operative diagnostic tool was 85%. CONCLUSION: Computed tomography angiogram is simple, quick and can provide an accurate pre-operative renal vascular anatomy in terms of number and location of renal vessels, early dividing renal arteries and crossing vessels at UPJ. PMID- 19120528 TI - Discarding antimicrobial prophylaxis for transurethral resection of bladder tumor: a feasibility study. AB - OBJECTIVES: To evaluate the feasibility of discarding antimicrobial prophylaxis (AMP) for transurethral resection of bladder tumor (TURBT). METHODS: One-hundred and sixty-two patients undergoing TURBT, with no risk factors for infectious complications were included in this prospective study between April 2006 and April 2008. Forty-four patients received single oral dose of 200 mg levofloxacin for AMP (LVFX group), and the remaining 118 received no AMP (no AMP group). Rates of postoperative infectious complications were compared between the two groups. RESULTS: There was no significant difference between the two groups in terms of baseline characteristics, including age, sex, history of TURBT, number of tumors, duration of operation and days of catheterization. Symptomatic urinary tract infections (UTIs) occurred in four patients (3.4%) in the no AMP group and one patient (2.3%) in the LVFX group (P = 0.61). All patients having UTIs were men, and were successfully treated by immediate administration of antibiotics. No patients developed extra-urinary tract infections. CONCLUSIONS: Based on our findings, AMP for TURBT in patients with no risk factors for infectious complications is not necessary. The use of antibiotics might be deferred until postoperative infections develop. PMID- 19120529 TI - Technical acquisition and dosimetric assessment of iodine-125 permanent brachytherapy in localized prostate cancer: our first series of 100 patients. AB - OBJECTIVES: To assess our initial experience in the treatment of localized prostate cancer using low-dose-rate brachytherapy (LDR-brachytherapy) with iodine 125. METHODS: One-hundred consecutive patients received LDR-brachytherapy between July 2004 and October 2006. Seventy-six patients were treated with seed implantation alone, whereas 24 patients were treated with a combination of brachytherapy and external beam radiotherapy. The minimal percentage of the dose received by 90% of the prostate gland (%D90), the percentage prostate volume receiving 100% of the prescribed minimal peripheral dose (V100), and the operation time were compared among every 10 consecutive patients. RESULTS: The means of %D90 and V100 were 109.6% and 93.4%, respectively. When compared with the first 10 patients, both D90 and V100 showed significant improvement in the following 10 consecutive patients. Similarly, the mean operation time decreased significantly according to the accumulated number of patients. CONCLUSIONS: Our initial experience with the first 100 cases suggests that LDR-brachytherapy needs accumulation of many more patients to obtain high-quality post-implant dosimetric outcomes. PMID- 19120530 TI - Current use of the artificial urinary sphincter and its long-term durability: a nationwide survey in Japan. AB - OBJECTIVES: Although the artificial urinary sphincter (AUS) is one of the most effective surgical treatments for severe urinary incontinence, little is known about its use in Japan. A nationwide survey was done to determine contemporary trends in AUS use and its long-term durability. METHODS: Data on AUS units sold in Japan were provided directly by Takai Hospital Supply Co., Ltd., Tokyo, Japan, and a survey form was sent to all 44 institutes where AUS implantation had been carried out. The survey included various demographic and preoperative variables, surgical variables, and postoperative outcomes. RESULTS: Between 1994 and 2007, a total of 100 AUS devices had been provided in Japan. Of the 44 institutes, 24 responded to the survey, and a total of 64 patients were enrolled in the study. Post-urological surgery incontinence accounted for 81.3% of the indications. During the mean follow-up of 50 months, mechanical failure occurred in four (6.2%), and the device was removed in 13 (20.3%) due to infection (14.0%), erosion (4.7%), or urination difficulty (1.5%). Of the 58 patients evaluated, 91.4% reported social continence. Five- and 10-year failure-free rates were 74.8% and 70.1%, respectively. On multivariate analysis, operative time was an independent predictor of treatment failure (P = 0.0334). CONCLUSIONS: Considering recent trends in prostate surgery, the AUS may be significantly underused in Japan. Although excellent long-term durability has been achieved, a learning effect appears to be evident. The Japanese urological community needs to provide appropriate patients with this treatment option. PMID- 19120531 TI - Renal ischemia time in laparoscopic surgery: an experimental study in a porcine model. AB - OBJECTIVES: Laparoscopic nephron sparing surgery is becoming more common, only limited by experience of the surgeon and the renal ischemia time. It is currently unknown if laparoscopic surgery alters the accepted 30 min threshold for renal ischemia. The purpose of this study was to define the normothermic ischemic threshold in the laparoscopic setting. METHODS: Twenty-four domestic female pigs underwent a laparoscopic transperitoneal procedure after randomization to one of five treatment groups: 30, 60, 75, 90, and 120 min of unilateral renal ischemia. One animal was used as a control specimen and was not exposed to any surgery or pneumoperitoneum. The contralateral kidney in each animal was used as an individual control for the corresponding time of pneumoperitoneum. The animals were euthanized after 3 weeks, when both renal units were harvested and data were analyzed. RESULTS: Three animals died secondary to complications from their procedure and were replaced. There was no association between urine volume and ischemic time, but urine creatinine and creatinine clearance decreased significantly as ischemic time increased. The control animal and 30 min ischemic group had similar results. Functional renal parameters decreased in those animals exposed to greater than 75 min of ischemia. The histologic analysis did not show any significant differences among the experimental groups. CONCLUSION: Renal function begins to deteriorate between 75 and 90 min of renal vascular occlusion in this two-renal unit laparoscopic porcine model. PMID- 19120532 TI - Incidence of inguinal hernia after prostate surgery: open radical retropubic prostatectomy versus open simple prostatectomy versus transurethral resection of the prostate. AB - Our objective was to determine the incidence of inguinal hernia (IH) after surgery for prostatic diseases. Medical records of 395 patients who underwent radical retropubic prostatectomy (RRP; n = 155), open simple prostatectomy (OP; n = 35), or transurethral resection of the prostate (TURP; n = 205) at the Chibaken Saiseikai Narashino Hospital from April 2000 to March 2007 were retrospectively evaluated. The incidence of IH was 23.9% in the RRP group, 18.9% in the OP group, and 2% in the TURP group. Overall, 91.9% in the RRP and 83.3% in the OP group developed an IH within 2 years postoperatively. The laterality of IH after open surgery was mainly on the right side. Subclinical IH were seen in 25% of RRP cases. The existence of subclinical IH was the only significant risk factor for postoperative IH in this analysis. Furthermore, OP and RRP procedures significantly increased the risk of postoperative IH compared with TURP. The hernia-free ratios were significantly lower after RRP and OP than after TURP (vs RRP: P < 0.001; vs OP: P < 0.001). Our findings confirm that a lower abdominal incision itself is associated with postoperative IH in patients undergoing prostate surgery. Attention must be paid to pre-existing subclinical IH through careful preoperative assessment. Patients should be followed for more than 2 years due to the high incidence of postoperative IH. PMID- 19120533 TI - Vitamin D for prevention of chronic disease: the need for continued research. PMID- 19120534 TI - Endoscopic ultrasound-guided fine-needle aspiration when combined with positron emission tomography improves specificity and overall diagnostic accuracy in unexplained mediastinal lymphadenopathy and staging of non-small-cell lung cancer. AB - BACKGROUND: The aim of this study was to assess the incremental value of endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) to positron emission tomography (PET) in the diagnosis of unexplained mediastinal lymphadenopathy and staging of non-small-cell lung cancer (NSCLC). METHODS: Patients who had both EUS-guided FNA and PET were retrospectively identified from an EUS database at a tertiary hospital. All EUS-guided FNA were carried out by one endoscopist between August 2002 and April 2005, either for the diagnosis of unexplained mediastinal lymphadenopathy or for the staging of NSCLC. Results of PET and EUS were compared with histology. A true histological positive result was defined as histological involvement in either surgery (mediastinoscopy or resection) or EUS-guided FNA. A true histological negative result was defined as negative involvement at surgery (mediastinoscopy or resection). RESULTS: Forty nine patients who had both PET scanning and EUS-guided FNA for diagnosis of unexplained mediastinal lymphadenopathy or staging of NSCLC were identified. Of these, 33 (73% males, n = 24, age range = 44-78 years, mean = 62 years) had surgical confirmation of mediastinal lymph node pathology. In these patients, PET alone showed sensitivity, 95%; specificity, 90%; positive predictive value, 87%; negative predictive value, 90% and accuracy, 88%; whereas the addition of EUS guided FNA increased the overall specificity and positive predictive value to 100%, with an overall accuracy of 97%. CONCLUSIONS: This study suggests that EUS guided FNA complements PET by improving the overall specificity and thereby the accuracy for diagnosis of unexplained mediastinal lymphadenopathy. It provides a minimally invasive technique to assess the mediastinum in patients with NSCLC and is particularly valuable in cases in which PET findings are equivocal. PMID- 19120535 TI - Chronic urticaria: the autoimmune paradigm. AB - Chronic urticaria is a disease consisting of spontaneous pruritic welts, present on all or most days for more than 6 weeks. It is commonly supposed to be allergic in origin, although allergy is not the cause in the majority of cases, and it has therefore been termed 'chronic idiopathic urticaria'. Recent evidence indicates that at least a subset of patients in whom no extrinsic or internal cause can be identified are in fact autoimmune in origin. This is based mainly on the detection of pathogenic autoantibodies to the high-affinity immunoglobulin E receptor FcepsilonR1, which are thought to activate cutaneous mast cells. In this article, we review the evidence that has given rise to this autoimmune 'paradigm' and its impact on diagnosis and management. PMID- 19120536 TI - Disease mongering: expanding the boundaries of treatable disease. AB - Traditionally, the promotional activities of medical industries have been product specific. In recent years, however, there have been examples where companies have worked through partnerships, which have included clinicians, to expand the boundaries of treatable disorders. The main motivation appears to be to increase sales of commercial products. The term 'disease mongering' has been applied to these activities. Whereas some disease awareness programmes may bring benefits in the form of improved recognition and management of disorders, the presence of strong commercial interests probably distorts the traditional processes by which treatable diseases have been defined. This can result in individual patients being exposed to potential harms, with little expectation of benefit and will place an unwarranted burden on the publicly funded health-care system. None of this can happen without the collaboration of the medical profession that needs to be aware of the risks of becoming involved in commercially supported 'consensus' groups that are reviewing the definition and management of diseases. PMID- 19120537 TI - Tako-tsubo-like cardiomyopathy after EpiPen administration. AB - Tako-tsubo-like cardiomyopathy is characterized by acute chest pain, electrocardiographic changes and increased cardiac enzymes in the absence of obstructive coronary vessel disease. We describe the development of tako-tsubo like cardiomyopathy in an elderly woman after the use of an EpiPen for generalized urticaria and angioedema. As adrenaline may participate in the pathogenesis of this condition, the need for careful patient selection and education in the use of adrenaline self-injectors remains imperative. PMID- 19120538 TI - Cerebral venous thrombosis associated with ulcerative colitis. AB - Thromboembolic complications, such as deep venous thrombosis and pulmonary embolism, are well recognized in patients with inflammatory bowel disease (IBD). We describe three cases of cerebral venous thrombosis complicating ulcerative colitis. Cerebral venous thrombosis is a rare but potentially devastating complication of IBD, and the diagnosis needs to be considered in any patient with IBD presenting with neurological symptoms. PMID- 19120539 TI - Charcot shoulder in syringomyelia. PMID- 19120540 TI - A perfect host. PMID- 19120541 TI - Drug fever due to labetalol. PMID- 19120542 TI - Restless leg syndrome, dopamine agonists and sleep attacks. PMID- 19120544 TI - Death as an 'adverse outcome'. PMID- 19120545 TI - Air pollution: the new cardiovascular risk factor. PMID- 19120546 TI - Expression of genes encoding kinin receptors in peripheral blood mononuclear cells from patients with acute coronary syndromes. AB - BACKGROUND: Inflammation plays a critical role in all stages of atherogenesis, including plaque destabilization leading to the rupture and local thrombosis, clinically manifested as unstable angina (UA) or myocardial infarction (MI). Recent data report enhanced expression of numerous pro-inflammatory genes in patients with acute coronary syndrome (ACS) both in plaque and in inflammatory cells. Kinins are peptides involved in vasodilation, vascular permeability, pain and inflammation. Their effects are mediated by two receptors, B1 and B2. As the role of kinins in ACS is not clear, the aim of the study was to assess the expression of the genes encoding kinin receptors in patients with ACS. METHODS: The study was carried out on 40 patients with ACS and 10 age-matched healthy subjects (control (C)). To evaluate gene expression of B1 and B2 kinin receptors, total mRNA was extracted from peripheral blood mononuclear cells and the number of mRNA copies was assessed by quantitative reverse transcriptase-polymerase chain reaction. RESULTS: In patients with MI and UA, the B1 receptor (B1R)/B2 receptor (B2R) ratio was inversed compared with healthy subjects (C group) (MI vs C: 1.54 +/- 0.39 vs 0.36 +/- 0.04; P < 0.01; UA vs C: 2.13 +/- 0.98 vs 0.36 +/- 0.04; P < 0.05 respectively). B2R gene mRNA level was markedly lower in MI group versus C group (24 216 +/- 5409 copies/microg vs 39 908 +/- 5309 copies/microg; P < 0.05). The difference in B1R gene expression between MI and C group was negligible. We have not observed differences in studied genes expression between UA and C groups. CONCLUSION: Patients with ACS show inverted B1R/B2R ratio. Such disturbance in kinin signalling may reflect increased activation of circulating mononuclears, which are important participants of atherosclerotic plaque development and eventually rupture. PMID- 19120547 TI - Retinal vein occlusion: an approach to diagnosis, systemic risk factors and management. AB - Retinal vein occlusion (RVO) is the second most common retinal vascular disease after diabetic retinopathy and is a common cause of visual morbidity and blindness in the elderly. A large proportion of patients with RVO have a history of cardiovascular disease, hypertension, diabetes mellitus or open-angle glaucoma. Although RVO is sometimes associated with thrombophilias and coagulation abnormalities, the role of coagulation factors in the development of RVO remains unclear. This review did not find strong evidence to support an extensive work-up for thrombophilic and coagulation diseases for the vast majority of patients. However, when tests for common cardiovascular risk factors for RVO are negative, evaluation for potential coagulation disorders may be indicated, particularly in young patients and in patients with bilateral RVO, a history of previous thromboses or a family history of thrombosis. PMID- 19120548 TI - Heart transplantation: who should be considered and when? AB - Heart transplantation is now firmly established as the most effective therapy for selected patients with end-stage heart disease who have exhausted all other treatment options. The excellent long-term survival and quality of life achieved by the large majority of heart transplant recipients has led to considerable broadening of the recipient eligibility criteria over the past 24 years of continuous transplant activity in Australia and New Zealand. Although these changes in recipient eligibility criteria have resulted in a marked increase in the potential pool of transplant recipients, heart transplantation remains a relatively rare procedure because of the scarcity of suitable cardiac donors. The primary aim of this review was to provide clinicians with guidelines that enable them to identify patients who are potential candidates for heart transplantation and those who are not. Appropriate timing of referral is also discussed. Apart from their role in the evaluation and management of patients referred for heart transplantation, cardiopulmonary transplant units provide a number of other important services. These include assistance in the diagnostic evaluation and management of patients with primary myocardial diseases and the investigation of novel alternative therapies for patients with end-stage heart disease. PMID- 19120549 TI - Arterial stiffness is associated with raised levels of the inflammatory marker erythrocyte sedimentation rate among ischaemic stroke patients. AB - We studied the relationship of arterial stiffness, measured by carotid-femoral pulse wave velocity and inflammation, measured by serum erythrocyte sedimentation rate among 334 ischaemic stroke patients. There was a significant correlation between carotid-femoral pulse wave velocity and erythrocyte sedimentation rate (P = 0.001), a relationship independent of age, hypertension, diabetes and smoking. Arterial stiffness and inflammation are associated among ischaemic stroke patients and are independent of established vascular risk factors. PMID- 19120550 TI - Decreasing mortality from non-Hodgkin lymphoma in Australia. AB - Based on the number of new cases (incidence), non-Hodgkin lymphoma is an increasingly common cancer in Australia and many developed countries. Until recently, mortality trends have been stable or slightly increasing. However, since the year 2000, mortality has decreased every year by an average of 5.1% per year (95% confidence interval (CI) -7.1 to -3.1%), whereas incidence has continued to increase at 0.9% per year (95%CI 0.6 to 1.2%). It was not possible with the population-based registry data available to us to untangle the causes of the decrease in mortality. The stable mortality rates during the 1990s (in the face of increasing incidence) might have been because of introduction of novel therapies such as autologous stem cell transplant for relapsed diffuse large cell lymphoma or the purine analogue-based therapy for indolent lymphomas. A plausible explanation for the large decrease in mortality since 2000 is the introduction of the monoclonal antibody rituximab. PMID- 19120552 TI - Chylous ascites. PMID- 19120553 TI - Spontaneous peroneal compartment syndrome causing acute foot drop. PMID- 19120554 TI - Indigenous health: a continuing litany of no improvement. PMID- 19120556 TI - Caution is advised in the treatment of chronic hyponatremia. PMID- 19120566 TI - Coerced medication in psychiatric care. PMID- 19120567 TI - Coerced medication in psychiatric inpatient care: literature review. AB - AIM: This paper is a report of a narrative review of the coercive use of medication in inpatient psychiatric care to identify a demographic and clinical profile of patients who are forcibly medicated, and to examine patient and staff views of this practice. BACKGROUND: Lack of compliance with medication is associated with quicker relapse and increased risk to self and others in mental disorder. It is this increased risk which provides the ethical and legal grounds for detaining and treating psychiatric patients without their consent. Legislation for involuntary psychiatric treatment exists in all European Union member states and in other western countries. DATA SOURCES: Online bibliographic databases from 1980 to 2008 were searched, including British Nursing Index, CINAHL, PsycINFO, EMBASE and MEDLINE. Search terms relating to coercion, force, chemical restraint, rapid tranquilization, inpatients and psychiatry were used. REVIEW METHODS: Titles and abstracts were reviewed. All peer reviewed papers concerning coercion in the administration of medication in inpatient psychiatric care were included and a narrative review was conducted. RESULTS: Fourteen papers from seven countries were included. Patients who have experience coerced medication tend to be aged in their 30s, with a diagnosis of schizophrenia, bipolar or other psychotic disorders, and are often involuntarily admitted. Assault or threat of assault is the main reason for giving forced medication. CONCLUSION: There is a dearth of literature in the area of coercion in administration of medication and much more research is needed examining all aspects of this contentious practice. PMID- 19120568 TI - Informed consent to healthcare interventions in people with learning disabilities -an integrative review. AB - AIM: This paper is a report of an integrative review of informed consent to healthcare interventions in people with learning disabilities. BACKGROUND: Consent to treatment lies at the heart of the relationship between patient and healthcare professional. In order for people with learning disabilities to have equity of access to health care, they need to be able to give informed consent to health interventions--or be assessed as incompetent to give consent. DATA SOURCES: The British Nursing Index (BNI), CINAHL, MEDLINE, Social Care Online, ERIC and ASSIA and PsycINFO databases were searched using the search terms: Consent or informed choice or capacity or consent to treat* or consent to examin* AND Learning disab* or intellectual* disab* or mental* retard* or learning difficult* or mental* handicap*. The search was limited to papers published in English from January 1990 to March 2007. REVIEW METHODS: An integrative review was conducted and the data analysed thematically. RESULTS: Twenty-two studies were reviewed. The main themes identified were: life experience, interaction between healthcare professionals and participants, ability to consent, and psychometric variables. A consensus seemed to emerge that capacity to consent is greater in people with higher cognitive ability and verbal skills, but that the attitudes and behaviour of healthcare professionals was also a crucial factor. CONCLUSION: The findings support use of the functional approach to assessing mental capacity for the purpose of obtaining informed consent. Future research into informed consent in people with learning disabilities is needed using real life situations rather than hypothetical vignettes. PMID- 19120569 TI - Hope, expectations and recovery from illness: a narrative synthesis of qualitative research. AB - AIM: This paper is a report of a narrative literature review conducted to explore how expectations and wants are distinguished in empirical research on hope and illness and the related issues of realistic hope and temporality. BACKGROUND: Particularized hope has been identified as comprising wants and expectations. The distinction is important in relation to debates around 'realistic' hopes, the temporal dimension of hope and hope-sustaining strategies. DATA SOURCES: A systematic search of the literature was undertaken for qualitative research papers published between January 1996 and July 2007 relating to hope and recovery in adults with physical ill health. Seven papers were identified. REVIEW METHODS: A narrative synthesis approach was adopted and the papers were appraised for quality using the Critical Appraisal Skills Programme method. Textual descriptions and tabulation were used to compare central themes and thematic analysis was used to explore the findings. RESULTS: A lack of conceptual clarity in relation to hope as a want or an expectation was identified. Healthcare professionals' role in encouraging 'realistic' hopes emerged as problematic because of the lack of understanding about the possible benefits of hope and difficulties in identifying 'realistic' hopes. There has been limited research exploring temporality and the impact of hope-sustaining activities. CONCLUSION: The lack of clarity about particularized hope, its dimensions, properties or different forms limits knowledge about the conditions under which hope is a positive force versus when it can be damaging. Distinguishing conceptually between hope as a want and hope as an expectation has potential value in improving healthcare practice and informing future investigations. PMID- 19120572 TI - Next-of-kin caregivers in palliative home care--from control to loss of control. AB - AIM: This paper is a report of a study to describe situations influencing next-of kin caregivers' ability to manage palliative care in the home. BACKGROUND: Palliative home care often involves major resources from next-of-kin, municipal and county personnel. Next-of-kin caregivers' situation is depicted as so demanding and exhausting that it can affect their mental health and limit their ability to continue as a caregiver. METHODS: A qualitative study using the Critical Incident Technique was conducted. Nine next-of-kin caregivers from palliative home care were strategically chosen and data were collected using audiotaped interviews in 2005. FINDINGS: The analysis resulted in two main areas: 'Maintaining control' and 'Losing control'. Next-of-kin caregivers wanted to maintain control over their lives by being continuously available 24 hours a day and by supporting and taking complete responsibility for all of the patient's needs. They lost this control when professional assistance was lacking and they described feelings of inadequacy when their physical energy or time was insufficient. They felt incapable as the patient's physical or mental persona metamorphosed or as serious symptoms developed that they could not control. CONCLUSION: Next-of-kin caregivers and the patients must feel free to choose where palliative home care will be provided and that their choice at any time can be re-evaluated, due to insecurity or workload changes. When planning for palliative care, they should be informed that having only one caregiver reduces the chances of completing palliative care in the home. PMID- 19120573 TI - Illness consequences after myocardial infarction: problems with physical functioning and return to work. AB - AIM: This paper is a report of a study to explore health problems, physical and mental functioning, and physical activity in working-age patients after myocardial infarction, in order to assess the possible effects of these factors on return to work. BACKGROUND: A diagnosis of myocardial infarction may discourage patients from continuing an active working life. Enabling myocardial infarction patients to return to work has benefits for both individuals and society. METHODS: A convenience sample was recruited of 88 patients, or =1 of the six antibiotic resistance gene(s). The tetA(P), tetB(P) and tet(M) genes were in 53%, 22% and 8%, and the erm(B), erm(Q) and mef(A) genes in 26%, 1% and 18% of the isolates, respectively. The mef(A) gene and flanking regions were sequenced. The tet(M), erm(B), erm(Q) and mef(A) genes transfer independently from C. perfringens donors to the Enterococcus faecalis recipient. CONCLUSIONS: Six resistance genes were found in the environmental C. perfringens with the most common being the tetA(P) gene and the erm(Q) gene the least common. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time conjugal transfer of macrolide resistance genes and/or the tet(M) gene from C. perfringens has been demonstrated. The data presented supports the hypothesis that antibiotic resistant environmental C. perfringens are capable of acting as reservoirs for these antibiotic resistance genes. PMID- 19120612 TI - Production and characterization of pure Clostridium spore suspensions. AB - AIMS: A general protocol was derived for optimizing the production of pure, high concentration Clostridium endospore suspensions. METHODS AND RESULTS: Two sporulation methods were developed that yielded high concentrations of notably pure Clostridium sporogenes, C. hungatei and C. GSA-1 (Greenland ice core isolate) spore suspensions (10 ml of 10(9) spores ml(-1) with >99% purity each). Each method was derived by evaluating combinations of three sporulation conditions, including freeze drying of inocula, heat shock treatment of cultures, and subsequent incubation at suboptimal temperatures that yielded the highest percentage of sporulation. Pure spore suspensions were characterized in terms of dipicolinic acid content, culturability, decimal reduction time (D) value for heat inactivation (100 degrees C) and hydrophobicity. CONCLUSIONS: While some Clostridium species produce a high percentage of spores with heat shock treatment and suboptimal temperature incubation, other species require the additional step of freeze drying the inocula to achieve a high percentage of sporulation. SIGNIFICANCE AND IMPACT OF THE STUDY: Pure Clostridium spore suspensions are required for investigating species of medical and environmental importance. Defining the conditions for optimal spore production also provides insight into the underlying mechanisms of Clostridium sporulation. PMID- 19120613 TI - Regional differences in bacterial flora in harbour porpoises from the North Atlantic: environmental effects? AB - AIMS: Microbiological findings in harbour porpoises from different regions of the North Atlantic were compared. Results in animals from the North and Baltic Seas were evaluated over a period of 18 years for changes in the microbiological flora. METHODS AND RESULTS: Microbiological investigations were performed on 1429 organ samples from the lung, liver, kidney, spleen, intestine, and mesenteric lymph nodes from harbour porpoises of the German North and Baltic Seas, Greenlandic, Icelandic and Norwegian waters. A large variety of bacteria, including potentially pathogenic bacteria like Brucella sp., Clostridium perfringens, Escherichia coli, Erysipelothrix rhusiopathiae, beta-haemolytic streptococci and Staphylococcus aureus were isolated. Those bacteria were associated with bronchopneumonia, gastroenteritis, hepatitis, pyelonephritis, myocarditis and septicemia. CONCLUSIONS: Organs from animals originating from Greenlandic and Icelandic waters showed clearly less bacterial growth and fewer associated pathological lesions compared to animals from the German North and Baltic Seas and Norwegian waters. SIGNIFICANCE AND IMPACT OF THE STUDY: Differences in bacterial findings and associated lesions between harbour porpoises from the German North and Baltic Seas and animals from Greenlandic, Norwegian and Icelandic waters may result from higher stress due to anthropogenic activities such as chemical pollutants in the North and Baltic Seas. PMID- 19120614 TI - Microbial characterization and quantification of an anaerobic sludge degrading dimethyl phthalate. AB - AIMS: Characterization and quantification of microbial community in dimethyl phthalate (DMP)-degrading anaerobic sludge using molecular techniques. METHODS AND RESULTS: An enriched anaerobic sludge effectively degrading over 99% of dimethyl phthalate in an upflow anaerobic sludge blanket (UASB) reactor for 530 days was characterized and quantified by 16S rRNA-based molecular methods. A total of 78 Bacteria clones were classified into 22 operational taxonomic units (OTUs) in nine divisions, including Firmicutes, Proteobacteria, Chloroflexi, Thermotogae, Bacteroidetes/Chlorobi, Spirochaetes, Acidobacteria and two candidate divisions. The two most abundant OTUs were likely responsible, respectively, for the de-esterification of DMP and the subsequent phthalate degradation. The outer layer of the granule was dominated by Bacteria; whereas the interior was by Archaea, of which 89 +/- 5% were acetoclastic Methanosaetaceae and 11 +/- 5% hydrogenotrophic Methanomicrobiales. CONCLUSIONS: Twenty-two Bacteria OTUs in DMP-degrading anaerobic sludge distributed in nine divisions. The two most abundant OTUs were likely responsible respectively for the de-esterification of DMP and the subsequent phthalate degradation. Layered granular microstructure of DMP-degrading anaerobic sludge suggested that the rate of DMP de-esterification is faster than its inward diffusion rate. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is the first study to characterize and quantify the microbial community in the anaerobic phthalic ester degrading sludge from the anaerobic reactor. PMID- 19120615 TI - Mycofumigation with Oxyporus latemarginatus EF069 for control of postharvest apple decay and Rhizoctonia root rot on moth orchid. AB - AIMS: To characterize the volatile antifungal compound produced by Oxyporus latemarginatus EF069 and to examine in vitro and in vivo fumigation activity of the fungus. METHODS AND RESULTS: An antifungal volatile-producing strain, O. latemarginatus EF069 inhibited the mycelial growth of Alternaria alternata, Botrytis cinerea, Colletotrichum gloeosporioides, Fusarium oxysporum f. sp. lycopersici, and Rhizoctonia solani by mycofumigation. An antifungal volatile compound was isolated from the hexane extract of wheat bran-rice hull cultures of O. latemarginatus EF069 by repeated silica gel column chromatography and identified as 5-pentyl-2-furaldehyde (PTF). The purified PTF inhibited mycelial growth of R. solani in a dose-dependent manner. The mycofumigation with solid cultures of EF069 also reduced effectively the development of postharvest apple decay caused by B. cinerea and Rhizoctonia root rot of moth orchid caused by R. solani. CONCLUSIONS: Oxyporus latemarginatus EF069 showed in vitro and in vivo fumigation activity against plant pathogenic fungi by producing 5-pentyl-2 furaldehyde. SIGNIFICANCE AND IMPACT OF THE STUDY: Oxyporus latemarginatus EF069 producing an antifungal volatile compound may be used as a biofumigant for the control of fungal plant diseases. PMID- 19120616 TI - Mixed aerobic and anaerobic microbial communities in benzene-contaminated groundwater. AB - AIMS: To investigate the factors affecting benzene biodegradation and microbial community composition in a contaminated aquifer. METHODS AND RESULTS: We identified the microbial community in groundwater samples from a benzene contaminated aquifer situated below a petrochemical plant. Eleven out of twelve groundwater samples with in situ dissolved oxygen concentrations between 0 and 2.57 mg l(-1) showed benzene degradation in aerobic microcosm experiments, whereas no degradation in anaerobic microcosms was observed. The lack of aerobic degradation in the remaining microcosm could be attributed to a pH of 12.1. Three groundwaters, examined by 16S rRNA gene clone libraries, with low in situ oxygen concentrations and high benzene levels, each had a different dominant aerobic (or denitrifying) population, either Pseudomonas, Polaromonas or Acidovorax species. These groundwaters also had syntrophic organisms, and aceticlastic methanogens were detected in two samples. The alkaline groundwater was dominated by organisms closely related to Hydrogenophaga. CONCLUSIONS: Results show that pH 12.1 is inimical to benzene biodegradation, and that oxygen concentrations below 0.03 mg l(-1) can support aerobic benzene-degrading communities. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings will help to guide the treatment of contaminated groundwaters, and raise questions about the extent to which aerobes and anaerobes may interact to effect benzene degradation. PMID- 19120617 TI - Transposition of IS10R in Lactococcus lactis. AB - AIMS: To characterize the transposition mechanism of the IS-element IS10R and study how this element is involved in gene disruption in Lactococcus lactis. METHODS AND RESULTS: The gene flciA confers immunity against lactococcin A in lactococci. However, the immunity function was lost when flciA was co-expressed with the regulator gene nisR on a plasmid in L. lactis NZ9000. By PCR and DNA sequencing, it was revealed that flciA in immune-negative transformants was disrupted by the IS-element IS10R. Such gene disruption did not occur when flciA was expressed alone nor when the plasmid-located nisR was mutated, suggesting that nisR is directly involved in the transposition. The sequence 5'-CACTTAACC 3', which was found in flciA and at both ends of the inserted IS10R, was identified as target site by site-directed mutagenesis. CONCLUSIONS: IS10R transposes in L. lactis NZ9000 in a nisR-dependent fashion and employs the sequence 5'-CACTTAACC-3' as integration site. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first time IS10R and aspects of its transposition are described in the industrial important bacterium L. lactis. The highly controllable insertion of IS10R into a target site might present a great potential as a gene disruption system. PMID- 19120618 TI - Modelling the number of viable vegetative cells of Bacillus cereus passing through the stomach. AB - AIMS: Model the number of viable vegetative cells of B. cereus surviving the gastric passage after experiments in simulated gastric conditions. MATERIALS AND METHODS: The inactivation of stationary and exponential phase vegetative cells of twelve different strains of Bacillus cereus, both mesophilic and psychrotrophic strains isolated from food and faeces from healthy and ill individuals, in simulated gastric conditions was determined using decimal reduction times at low pH (D(pH)). Subsequently inactivation rates were calculated. Inclusion of the inactivation rates into models describing the course of the gastric pH after the consumption of meal of solid food and the transfer of food from the stomach to the small intestine resulted in numbers of viable Bacillus cereus vegetative cells able to pass the stomach. CONCLUSIONS: According to the model, 3-26% of the ingested vegetative cells from Bacillus cereus may survive the gastric passage, dependent on the growth phase of the vegetative cells, the type of strains, and the age of the consumer. SIGNIFICANCE AND IMPACT OF THE STUDY: Vegetative cells of Bacillus cereus may be involved in the onset of diarrhoeal disease to a greater extent than expected since up to 26% of the ingested cells survive simulated gastric conditions. PMID- 19120619 TI - Selection of Trichoderma strains capable of increasing laccase production by Pleurotus ostreatus and Agaricus bisporus in dual cultures. AB - AIMS: To select Trichoderma strains for enhanced laccase production in Pleurotus ostreatus or Agaricus bisporus cultures. METHODS AND RESULTS: Laccase production by P. ostreatus and A. bisporus was evaluated in liquid (axenic) and solid (dual cultures) malt extract medium. Oxidation of ABTS, DMP and syringaldazine was evaluated in order to assess the potential of Trichoderma strains to enhance laccase production by basidiomycetes. Selected Pleurotus-Trichoderma interactions yielded higher increases in laccase volumetric activity and an additional laccase isoform was produced. By contrast, Agaricus-Trichoderma interactions lead to smaller increases on laccase volumetric activity, probably as result of repression (or degradation) towards one of the laccases isoforms. CONCLUSIONS: The strains of P. ostreatus and A. bisporus assessed in this work showed good potential as laccase producers. The Trichoderma-mediated biological stimulation of laccase production by P. ostreatus and A. bisporus is relevant in order to develop highly productive processes. SIGNIFICANCE AND IMPACT OF THE STUDY: Extracellular laccases from basidiomycetes are produced only in small amounts. It is therefore important to increase process productivity for potential industrial applications. The results from this study enable the selection Trichoderma strains capable of increasing laccase production by P. ostreatus or A. bisporus in dual cultures. PMID- 19120620 TI - Design and evaluation of PCR primers which differentiate Escherichia coli O157:H7 and related serotypes. AB - AIMS: To develop methods to differentiate Escherichia coli O157:H7 and related serotypes by the use of amplicon length polymorphism (ALP) analysis based on identifying DNA sequence deletions within highly homologous regions of three sequenced E. coli strains. METHODS AND RESULTS: Potential primer locations along the ancestral genomic backbone were identified and evaluated against three sequenced genomes and then applied to a reference set of pathogenic E. coli strains. All 16 primer combinations generated the expected diagnostic fragments as predicted for the E. coli K12 MG1655, O157:H7 EDL933, and O157:H7B Sakai genomes. CONCLUSIONS: This study defines a collection of primers distributed along the length of the E. coli genome that were applied to ALP analysis methods to successfully differentiate between serotypes of E. coli O157:H7 and other E. coli serotypes. SIGNIFICANCE AND IMPACT OF THE STUDY: ALP-PCR analysis method was validated as an independent method of classification when compared with that of rep-PCR. The principles underlying ALP analysis can be readily applied for the detection and differentiation of other closely related microbial species because of the abundance of complete DNA sequence data for a large number of microbial genomes. PMID- 19120621 TI - Bacterial community dynamics during reduction of odorous compounds in aerated pig manure slurry. AB - AIMS: To study the microbial community responsible for the reduction of the polluting load during aerobic digestion of pig slurry. METHODS AND RESULTS: We analysed bacterial succession by nonculture-based methods and determined the physicochemical parameters and polluting substances during 6 days of aerobic digestion. The bacterial subpopulations evolved by aeration, predominantly Bacillus spp., degraded organic matter and vigorously consumed oxygen, as indicated by low oxidation-reduction potential (ORP). In this phase, the volatile fatty acid (VFA) levels drastically decreased, and VFAs were almost depleted on day 4. Simultaneously, the ammonia concentration decreased to its lowest level on day 4; thereafter, it increased until the end of the process. After the decrease in the total organic carbon content in the supernatant of the decomposed slurry, the ORP increased (approximately 0 mV), and the microbial community showed an abundance of lineages belonging to the phylum Proteobacteria. CONCLUSIONS: Bacillus was the predominant member of the bacterial community driving the VFA removal process. Their predominance was related to the presence of available carbon, including VFAs and changes in ORP. SIGNIFICANCE AND IMPACT OF THE STUDY: Information on the relationships among the involved microbes, polluting materials and physicochemical parameters will aid process design and retrofitting of the process. PMID- 19120622 TI - Detailed studies on Quercus infectoria Olivier (nutgalls) as an alternative treatment for methicillin-resistant Staphylococcus aureus infections. AB - AIMS: To investigate the antimethicillin-resistant Staphylococcus aureus (MRSA) mechanism of Quercus infectoria (nutgalls) extract and its components. METHODS AND RESULTS: Ethanol extract, an ethyl acetate fraction I, gallic acid and tannic acid could inhibit the growth of clinically isolated MRSA strains with minimum inhibitory concentration values between 63 and 250 microg ml(-1). Clumps of partly divided cocci with thickened cell wall were observed by transmission electron microscopy in the cultures of MRSA incubated in the presence of the ethanol extract, the ethyl acetate fraction I and tannic acid. Because cell wall structure of the organism structures seemed to be a possible site for antibacterial mechanisms, their effect with representative beta-lactam antibiotics were determined. Synergistic effects with fractional inhibitory concentration index ranged from 0.24 to 0.37 were observed with 76% and 53% of the tested strains for the combination of the ethanol extract with amoxicillin and penicillin G, respectively. CONCLUSIONS: The appearance of pseudomulticellular bacteria in the treated cells and the synergistic effect of the plant extract with beta-lactamase-susceptible penicillins suggest that the extract may interfere with staphylococcal enzymes including autolysins and beta lactamase. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results provide scientific data on the use of the nutgalls, which contain mainly tannin contents up to 70% for the treatment of staphylococcal infections. PMID- 19120623 TI - Identification and tracing of Enterococcus spp. by RAPD-PCR in traditional fermented sausages and meat environment. AB - AIMS: Four local small-scale factories were studied to determine the sources of enterococci in traditional fermented sausages. METHODS AND RESULTS: Different points during the production of a traditional fermented sausage type (fuet) were evaluated. Randomly amplified polymorphic DNA (RAPD)-PCR was used to type 596 Enterococcus isolates from the final products, the initial meat batter, the casing, the workers' hands and the equipment. Species-specific PCR-multiplex and the partial sequencing of atpA gene and 16S rRNA gene sequencing allowed the identification of the isolates: Enterococcus faecalis (31.4%), Enterococcus faecium (30.7%), Enterococcus sanguinicola (14.9%), Enterococcus devriesei (9.7%), Enterococcus malodoratus (7.2%), Enterococcus gilvus (1.0%), Enterococcus gallinarum (1.3%), Enterococcus casseliflavus (3.4%), Enterococcus hermanniensis (0.2%), and Enterococcus durans (0.2%). A total of 92 different RAPD-PCR profiles were distributed among the different factories and samples evaluated. Most of the genotypes found in fuet samples were traced back to their source. CONCLUSIONS: The major sources of enterococci in the traditional fermented sausages studied were mainly the equipment followed by the raw ingredients, although a low proportion was traced back to human origin. SIGNIFICANCE AND IMPACT OF THE STUDY: This work contributes to determine the source of enterococcal contamination in fermented sausages and also to the knowledge of the meat environment. PMID- 19120625 TI - Phyllosphere microbiology with special reference to diversity and plant genotype. AB - The phyllosphere represents the habitat provided by the aboveground parts of plants, and on a global scale supports a large and complex microbial community. Microbial interactions in the phyllosphere can affect the fitness of plants in natural communities, the productivity of agricultural crops, and the safety of horticultural produce for human consumption. The structure of phyllosphere communities reflects immigration, survival and growth of microbial colonists, which is influenced by numerous environmental factors in addition to leaf physico chemical properties. The recent use of culture-independent techniques has demonstrated considerable previously unrecognized diversity in phyllosphere bacterial communities. Furthermore, there is significant recent evidence that plant genotype can play a major role in determining the structure of phyllosphere microbial communities. The main aims of this review are: (i) to discuss the diversity of phyllosphere microbial populations; (ii) to consider the processes by which microbes colonize the phyllosphere; (iii) to address the leaf characteristics and environmental factors that determine the survival and growth of colonists; (iv) to discuss microbial adaptations that allow establishment in the phyllosphere habitat and (v) to evaluate evidence for plant genotypic control of phyllosphere communities. Finally, we suggest approaches and priority areas for future research on phyllosphere microbiology. PMID- 19120624 TI - Plant growth promotion and biological control of Pythium aphanidermatum, a pathogen of cucumber, by endophytic actinomycetes. AB - AIMS: To evaluate the potential of Actinoplanes campanulatus, Micromonospora chalcea and Streptomyces spiralis endophytic in cucumber roots, to promote plant growth and to protect seedlings and mature plants of cucumber from diseases caused by Pythium aphanidermatum, under greenhouse conditions. METHODS AND RESULTS: Three endophytic isolates, out of 29, were selected through tests aimed at understanding their mechanisms of action as biocontrol agents and plant growth promoters. When applied individually or in combination, they significantly promoted plant growth and reduced damping-off and crown and root rot of cucumber. The combination of the three isolates resulted in significantly better suppression of diseases and plant growth promotion, than where the plants were exposed to individual strains. CONCLUSIONS: The three selected actinomycete isolates colonized cucumber roots endophytically for 8 weeks, promoted plant growth and suppressed pathogenic activities of P. aphanidermatum on seedling and mature cucumber plants. SIGNIFICANCE AND IMPACT OF THE STUDY: The results clearly show that the endophytic, glucanase-producing actinomycetes used, especially as a combined treatment, could replace metalaxyl, which is the currently recommended fungicide for Pythium diseases in the United Arab Emirates. These endophytic isolates also have the potential to perform as plant growth promoters, which is a useful attribute for crop production in nutrient impoverished soils. PMID- 19120626 TI - Effects of anthrax lethal toxin on human primary keratinocytes. AB - AIMS: To investigate the effects of anthrax lethal toxin (LeTx) on human primary keratinocytes. METHODS AND RESULTS: We show here that human primary keratinocytes are resistant to LeTx-triggered cytotoxicity. All but one of the MEKs (mitogen activated protein kinase kinases) are cleaved within 3 h, and the cleavage of MEKs in keratinocytes leads to their subsequent proteasome-mediated degradation at different rates. Moreover, LeTx reduced the concentration of several cytokines except RANTES in culture. CONCLUSIONS: Our results indicate that primary keratinocytes are resistant to LeTx cytotoxicity, and MEK cleavage does not correlate with LeTx cytotoxicity. Although LeTx is considered as an anti inflammatory agent, it upregulates RANTES. SIGNIFICANCE AND IMPACT OF THE STUDY: According to a current view, the action of LeTx results in downregulation of the inflammatory response, as evidenced by diminished expression of several inflammatory biomarkers. Paradoxically, LeTx has been reported to attract neutrophils to cutaneous infection sites. This paper, which shows that RANTES, a chemoattractant for immune cells, is upregulated after exposure of keratinocytes to LeTx, although a number of other markers of the inflammatory response are downregulated. Our results might explain why the exposure of keratinocytes to LeTx results in the recruitment of neutrophils to cutaneous infection sites, while the expression of several inflammatory biomarkers is diminished. PMID- 19120627 TI - Engineered Saccharomyces cerevisiae that produces 1,3-propanediol from D-glucose. AB - AIMS: Saccharomyces cerevisiae is a safe micro-organism used in fermentation industry. 1,3-Propanediol is an important chemical widely used in polymer production, but its availability is being restricted owing to its expensively chemical synthesis. The aim of this study is to engineer a S. cerevisiae strain that can produce 1,3-propanediol at low cost. METHODS AND RESULTS: By using D glucose as a feedstock, S. cerevisiae could produce glycerol, but not 1,3 propanediol. In this study, we have cloned two genes yqhD and dhaB required for the production of 1,3-propanediol from glycerol, and integrated them into the chromosome of S. cerevisiae W303-1A by Agrobacterium tumefaciens-mediated transformation. Both genes yqhD and dhaB functioned in the engineered S. cerevisiae and led to the production of 1,3-propanediol from D-glucose. CONCLUSION: Saccharomyces cerevisiae can be engineered to produce 1,3-propanediol from low-cost feedstock D-glucose. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report on developing S. cerevisiae to produce 1,3 propanediol by using A. tumefaciens-mediated transformation. This study might lead to a safe and cost-efficient method for industrial production of 1,3 propanediol. PMID- 19120628 TI - Azoxystrobin and soil interactions: degradation and impact on soil bacterial and fungal communities. AB - AIMS: To provide an independent assessment of azoxystrobin effects on nontarget soil bacteria and fungi and generate some baseline information on azoxystrobin's persistence in soil. METHODS AND RESULTS: Plate based assay showed that azoxystrobin exhibited differential toxicity upon cultured fungi at different application rates. While (14)C labelled isotopes experiments showed that less than 1% of azoxystrobin was mineralized, degradation studies revealed over 60% azoxystrobin breakdown over 21 days. PCR DGGE analysis of 16S and 18S rRNA genes from different soil microcosms showed that azoxystrobin had some effects on fungal community after 21 days (up to 84 days) of incubation in either light or dark soil microcosms. Light incubations increased fungal diversity while dark incubations reduced fungal diversity. Bacterial diversity was unaffected. CONCLUSIONS: Significant biotic breakdown of parent azoxystrobin occurred within 21 days even in the absence of light. Azoxystrobin under certain conditions can reduce fungal soil diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: One of the few independent assessments of azoxystrobin (a widely used strobilurins fungicide) effects on soil fungi when used at the recommended rate. Azoxystrobin and metabolites may persist after 21 days and affect soil fungi. PMID- 19120629 TI - New cell-based indicator assays for the detection of human cytomegalovirus infection and screening of inhibitors of viral immediate-early 2 protein activity. AB - AIMS: Expression of early (E) genes of human cytomegalovirus (HCMV) is stimulated cooperatively by the activities of host cell transcription factors and the viral immediate-early 2 (IE2) protein. Taking advantage of the IE2-dependent inducibility of E gene promoters, in this study, we generated cell-based assays in which the expression of the enhanced green fluorescence protein (EGFP) reporter gene was driven by the UL54 or UL112/113 E promoters. METHODS AND RESULTS: Cell clones derived from a stably transfected human cell line permissive to HCMV replication showed a specific and inducible dose- and time-dependent EGFP response to HCMV infection. The sensitivity of these indicator cells for detecting infectious particles of clinical isolates of HCMV was comparable to that of a conventional plaque assay. The HCMV-induced EGFP expression was completely prevented by treatment of indicator cells with fomivirsen, an antisense oligodeoxynucleotide designed to block IE2 expression, and this inhibitory activity was also observed when the IE2 protein alone was constitutively expressed in EGFP indicator cells. CONCLUSIONS: The EGFP-based cell assays have proved to be a rapid, sensitive, quantitative and specific system for detection of HCMV and selection of antivirals. SIGNIFICANCE AND IMPACT OF THE STUDY: These new cell-based assays can be exploited as functional assays to detect infectious HCMV particles, as well as to screen antiviral compounds that interfere with IE2 activity. PMID- 19120630 TI - Effects of organic matter on acidic electrolysed water for reduction of foodborne pathogens on lettuce and spinach. AB - AIMS: To evaluate the efficacy of acidic electrolysed water (EW) in the presence of organic matter (bovine serum) on the inoculated surfaces of lettuce and spinach. MATERIALS AND RESULTS: Lettuce and spinach leaves were inoculated with a cocktail of three strains each of Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes and treated with deionized water, acidic EW and acidic EW containing bovine serum (5, 10, 15 and 20 ml l(-1)) for 15 s, 30 s, 1 min, 3 min and 5 min at room temperature (22 +/- 2 degrees C). In the absence of bovine serum, acidic EW treatment reduced levels of cells below the detection limit (0.7 log) in 5 min. In the presence of bovine serum, bactericidal activity of acidic EW decreased with increasing serum concentration. CONCLUSIONS: Organic matter reduces the effectiveness of acidic EW for reducing pathogens on the surfaces of lettuce and spinach. SIGNIFICANCE AND IMPACT OF THE STUDY: From a practical standpoint, organic matter reduces the efficacy of acidic EW. This study was conducted to confirm the effect of organic matter on the properties of acidic EW in the inactivation of foodborne pathogens on the surface of vegetables. PMID- 19120631 TI - Auxin-treatment induces recovery of phytoplasma-infected periwinkle. AB - AIMS: To test the effect of auxin-treatment on plant pathogenic phytoplasmas and phytoplasma-infected host. METHODS AND RESULTS: In vitro grown periwinkle shoots infected with different 'Candidatus Phytoplasma' species were treated with indole 3-acetic acid (IAA) or indole-3-butyric acid (IBA). Both auxins induced recovery of phytoplasma-infected periwinkle shoots, but IBA was more effective. The time period and concentration of the auxin needed to induce recovery was dependent on the 'Candidatus Phytoplasma' species and the type of auxin. Two 'Candidatus Phytoplasma' species, 'Ca. P. pruni' (strain KVI, clover phyllody from Italy) and 'Ca. P. asteris' (strain HYDB, hydrangea phyllody), were susceptible to auxin treatment and undetected by nested PCR or detected only in the second nested PCR in the host tissue. 'Ca. P. solani' (strain SA-I, grapevine yellows) persisted in the host tissue despite the obvious recovery of the host plant and was always detected in the direct PCR. CONCLUSIONS: Both auxins induced recovery of phytoplasma-infected plants and affected tested 'Candidatus Phytoplasma' species in the same manner, implying that the mechanism involved in phytoplasma elimination/survival is common to both, IAA and IBA. SIGNIFICANCE AND IMPACT OF THE STUDY: The results imply that in the case of some 'Candidatus Phytoplasma' species, IBA-treatment could be used to eliminate phytoplasmas from in vitro grown Catharanthus roseus shoots. PMID- 19120632 TI - Archaea diversity within a commercial biogas plant utilizing herbal biomass determined by 16S rDNA and mcrA analysis. AB - AIMS: The Archaea diversity was evaluated in an agricultural biogas plant supplied with cattle liquid manure and maize silage under mesophilic conditions. METHODS AND RESULTS: Two different genes (16S rRNA; methyl-coenzyme-M-reductase, MCR) targeted by three different PCR primer sets were selected and used for the construction of three clone libraries comprising between 104 and 118 clones. The clone libraries were analysed by restriction fragment polymorphism (RFLP). Between 11 and 31 operational taxonomic units (OTUs) were detected and assigned to orders Methanomicrobiales, Methanosarcinales and Methanobacteriales. Over 70% of all Archaea OTUs belong to the order Methanomicrobiales which mostly include hydrogenotrophic methanogens. Acetotrophic methanogens were detected in minor rates. Similar relative values were obtained by a quantitative real-time PCR analysis. CONCLUSIONS: The results implied that in this biogas plant the most of the methane formation resulted from the conversion of H(2) and CO(2). SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports, for the first time, a molecular analysis of the archaeal community in this type of agricultural biogas plants. Therein the hydrogenotrophic methanogenesis seems to be the major pathway of methane formation. These results are in contrast with the common thesis that in biogas fermentations the primary substrate for methanogenesis is acetate. PMID- 19120633 TI - Isolation of highly copper-tolerant fungi from the smelter of the Naganobori copper mine, an historic mine in Japan. AB - AIMS: Copper is a critical metal of modern industry, and is the most widespread heavy metal contaminant in wastewater. Therefore, isolation of copper-tolerant microbes having the potential as biosorbent is fascinating not only from an environmental microbiology, but also from a biotechnology view point. In this study, we attempted to isolate highly copper-tolerant microbes from soil samples of the Nabanobori copper mine, the oldest mine in Japan. METHODS AND RESULTS: As a result of an enrichment culture, two fungal strains were isolated from soil of the smelter remains. The isolates could grow in a maximum of 200 mmol l(-l) Cu(2+), and grew under a wide pH range. The Cu(2+)-binding capacity of nontreated biomass of the isolates was around 35 mg Cu(2+) g(-1)-biomass. Analysis of 18S rDNA suggested that the isolates belong to the Aspergillus/Penicillium clade, but they represented a distinct lineage against known neighbours. CONCLUSION: The isolates were highly copper-tolerant, and their Cu(2+)-binding capacity was comparable to well-studied fungal sorbents. The isolates were implied as novel species. Soil of the historic old mine under weather-beaten conditions might be a suitable source for metal-tolerant microbes. SIGNIFICANCE AND IMPACT OF THE STUDY: The present results advance our understanding of metal-tolerant microbes, and offer a new tool for both environmental control and metal recovery operations. PMID- 19120634 TI - Phytase production by Sporotrichum thermophile in a cost-effective cane molasses medium in submerged fermentation and its application in bread. AB - AIMS: Phytase production by Sporotrichum thermophile in a cost-effective cane molasses medium in submerged fermentation and its application in bread. METHODS AND RESULTS: The production of phytase by a thermophilic mould S. thermophile was investigated using free and immobilized conidiospores in cane molasses medium in shake flasks, and stirred tank and air-lift fermenters. Among surfactants tested, Tweens (Tween-20, 40 and 80) and sodium oleate increased phytase accumulation, whereas SDS and Triton X-100 inhibited the enzyme production. The mould produced phytase optimally at a(w) 0.95, and it declined sharply below this a(w) value. The enzyme production was comparable in air-lift and stirred tank reactors with a marked reduction in fermentation time. Among the matrices tried, Ca-alginate was the best for conidiospore immobilization, and fungus secreted sustained levels of enzyme titres over five cycles. The phytic acid in the dough was efficiently hydrolysed by the enzyme accompanied by the liberation of soluble phosphate in the bread. CONCLUSIONS: The phytase production by S. thermophile was enhanced in the presence of Tween-80 in cane molasses medium. A peak in enzyme production was attained in 48 h in the fermenter when compared with that of 96 h in shake flasks. Ca-alginate immobilized conidiospores germinated to produce fungal growth that secreted sustained levels of phytase over five cycles. The bread made with phytase contained reduced level of phytic acid and a high-soluble phosphate. SIGNIFICANCE AND IMPACT OF THE STUDY: The phytase accumulation by S. thermophile was increased by the surfactants. The sustainability of enzyme production in stirred tank and air-lift fermenters suggested the possibility for scaling up of phytase. The bread made with phytase contained low level of antinutrient, i.e. phytic acid. PMID- 19120635 TI - Characterization of an acquired dps-containing gene island in the lactic acid bacterium Oenococcus oeni. AB - AIMS: To identify novel actors responsible for the marked adaptation of the Oenococcus oeni species to its environment. METHODS AND RESULTS: Genomic surveillance of the available genome sequences from O. oeni indicated the presence of a small ORF, encoding a protein named Dps(A). The cloned gene complemented the dps(-) mutant of Escherichia coli and conferred resistance to hydrogen peroxide, wine, and metals. The dps(A) gene was flanked by IS-related elements. The entire region was characterized by an anomalously high GC content compared to those reported for oenococcal genomes. The dps(A) gene was present in 15 of the 38 tested isolates. Positive strains originated from different geographical areas and sources. No change in tolerance to wine or to oxidative stress was observed between O. oeni strains harbouring dps(A) and those not harbouring this gene. CONCLUSIONS: Some O. oeni have acquired a functional homologue to the dps gene from E. coli as part of a mobile element. SIGNIFICANCE AND IMPACT OF THE STUDY: Dps(A) probably increases the bacterial fitness in response to environmental challenges. However, the physiological condition under which it adds a selective advantage to O. oeni during winemaking remains to be found. PMID- 19120636 TI - Comparative hydrocarbon utilization by hydrophobic and hydrophilic variants of Pseudomonas aeruginosa. AB - AIMS: To investigate hydrocarbon degradation by hydrophobic, hydrophilic and parental strains of Pseudomonas aeruginosa. METHODS AND RESULTS: Partitioning of hydrocarbon-degrading P. aeruginosa strain in a solvent/aqueous system yielded hydrophobic and hydrophilic fractions. Exhaustive partitioning of aqueous-phase cells yielded the hydrophilic variants (L), while sequential fractionation of the hydrophobic phase cells yielded successive fractions exhibiting increasing cell surface hydrophobicity (CSH). In hydrocarbon adherence assays (bacterial attachment to hydrocarbon), L had a value of 20%, which increased from 61.7% in first hydrophobic fraction (H(1)) to 72.2% in the third (H(3)). Crude oil degradation by L was 70%, but increased from 82% in H(1) to 93% in H(3). L variant produced most exopolysaccharides and reduced surface tension from about 73 to 49 mN m(-1). Rhamnolipid production was highest in L, but was not detected in all crude oil cultures. CONCLUSIONS: Hydrophobic subpopulations of hydrocarbon degrading P. aeruginosa exhibited greater hydrocarbon-utilizing ability than hydrophilic ones, or the parental strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Results demonstrate that a population of P. aeruginosa consists of cells with different CSH which affect hydrocarbon utilization. This potentially provides the population with the capacity to utilize different hydrophobic substrates found in petroleum. Judicious selection of such hydrophobic subpopulations can enhance hydrocarbon pollution bioremediation. PMID- 19120637 TI - An antifungal protein from Bacillus amyloliquefaciens. AB - AIMS: To isolate and characterize an antifungal protein from the culture broth of the bacterium Bacillus amyloliquefaciens. METHODS AND RESULTS: The antifungal protein designated as baciamin was isolated and exhibited a molecular mass around 50 kDa. Baciamin manifested a broad spectrum of antifungal activity. Baciamin could induce membrane permeabilization of tested fungi. Its antifungal activity was retained after incubation with trypsin and EDTA. Various ions tested did not affect its antifungal activity. Baciamin reduced the activity of HIV-1 reverse transcriptase (RT). It also inhibited proliferation of hepatoma, breast cancer and colon cancer cell lines. Baciamin augmented nitric oxide production by mouse macrophages. CONCLUSIONS: Bacillus amyloliquefaciens produces a broad-spectrum antifungal protein, baciamin. It induces membrane permeabilization in fungi but not in rabbit erythrocytes. Its antifungal activity is relatively thermostable, pH- and trypsin-stable. It demonstrates antiproliferative activity towards various tumour cells, nitric oxide-inducing activity towards macrophages, and inhibitory activity towards HIV-1 RT. SIGNIFICANCE AND IMPACT OF THE STUDY: Baciamin represents one of the few bacterial antifungal proteins reported to date. Most of the previously isolated antifungal molecules of bacterial origin are either peptides or ring compounds. Baciamin also exhibits other exploitable activities such as antitumour and immuno-enhancing activities. PMID- 19120638 TI - Isolation of Saccharomyces cerevisiae strains from different food matrices and their preliminary selection for a potential use as probiotics. AB - AIMS: To isolate acid- and bile-resistant Saccharomyces cerevisiae strains directly from food samples and to preliminarily select them on the basis of fundamental probiotic properties. METHODS AND RESULTS: A rapid screening method allowed the isolation and selection of 20 acid- and bile-resistant yeasts from foods, avoiding time-consuming isolation steps. The strains were characterized for their specific survival in simulated gastric juice and in intestinal fluid after pre-exposure at low pH. Ten isolates demonstrated a satisfactory survival percentage in intestinal fluid after pre-exposure to gastric juice and appreciable lipolytic and proteolytic properties, as demonstrated by the API-ZYM test. By using molecular methods five strains were identified as Saccharomyces cerevisiae, three as Candida spp., one as Candida pararugosa and one as Pichia spp. The Saccharomyces cerevisiae strains showed considerable probiotic properties, achieving a 80< % <90 survival through the simulated gastrointestinal tract, as well as interesting glucosidase activities. CONCLUSIONS: The research represents an efficient strategy to select and identify Saccharomyces cerevisiae strains with desirable acid and bile resistances. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper reports the direct selection of potentially probiotic yeasts from foods and provides indications about the ability of Saccharomyces cerevisiae strains to survive conditions simulating the human gastrointestinal tract. PMID- 19120639 TI - Characterization of starter lactic acid bacteria from the Finnish fermented milk product viili. AB - AIMS: Phenotypic and molecular methods were used to identify and compare the strain composition of three industrial dairy starters used for the manufacture of viili. METHODS AND RESULTS: Preliminary differentiation was made by phenotypic methods. Genotypic differentiation was carried out using polymerase chain reaction (PCR) and further characterization at strain level by pulsed-field gel electrophoresis (PFGE). The isolates could be assigned as acid-producing Lactococcus lactis strains of both lactis and cremoris subspecies, and aroma producers, identified as L. lactis subsp. lactis biovar diacetylactis and Leuconostoc mesenteroides. PCR analysis discriminated between the lactococcal subspecies, and cluster analysis of the digestion patterns of PFGE analysis revealed different genotypes in each subspecies. Each Leuconostoc-genotype seemed to be specific to only a single starter mix. CONCLUSIONS: The work proved that in addition to L. lactis subsp. lactis biovar diacetylactis and Leuc. mesenteroides subsp. cremoris, commercial viili starters of traditional origin may contain (i) only L. lactis subsp. cremoris, (ii) both L. lactis subsp. cremoris and L. lactis subsp. lactis as a minority, and - as a new discovery - (iii) only L. lactis subsp. lactis. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained give an overview of the microbial population of viili starters and can be exploited in the development of optimized starter cultures for industrial-scale manufacture of viili. PMID- 19120640 TI - Identification and characterization of the two-component cell lysis cassette encoded by temperate bacteriophage phiPYB5 of Lactobacillus fermentum. AB - AIMS: To characterize the two-component cell lysis cassette comprised of holin (Hyb5) and endolysin (Lyb5) encoded by Lactobacillus fermentum temperate bacteriophage phiPYB5, and illustrate the potential application of Lyb5 as therapeutic agents. METHODS AND RESULTS: The hyb5-lyb5 cassette was cloned from the genome library of phiPYB5, and the hyb5, lyb5 and hyb5-lyb5 cassette were expressed in E. coli BL21, respectively. The molecular weight of Hyb5 indicated by SDS-PAGE was 19 kDa, and Lyb5 was 45 kDa. Both Hyb5 and Lyb5 protein could induce cell lysis alone, resulting in the leakage of beta-galactosidase. However, the Hyb5-Lyb5 cassette lysed the host cells more rapidly and extensively. By zymogram analysis, Lyb5 exhibited a broad lytic spectrum. CONCLUSIONS: Overexpression of hyb5, lyb5 and hyb5-lyb5 cassette were carried out in E. coli and Lyb5 exhibited a broad lytic spectrum. SIGNIFICANCE AND IMPACT OF THE STUDY: The Lyb5 produced in E. coli exhibited a broad lytic spectrum against Gram positive strains including Staphylococcus aureus as well as Gram-negative strains such as Salmonella typhi, suggesting that Lyb5 provides a potential alternative of diagnostic tools and therapeutic agents. PMID- 19120641 TI - Occurrence and distribution of culturable enteroviruses in wastewater and surface waters of north-eastern Spain. AB - AIMS: Update information regarding occurrence and levels of culturable enteroviruses in several types of surface polluted waters in north-eastern Spain and determine the proportion of the different species and serotypes. METHODS AND RESULTS: The best procedures on hand in our laboratory for concentrating and quantifying culturable enteroviruses from different water sample types were used. Sequencing was used for typing the virus isolates. Geometric means of enteroviruses densities expressed in plaque forming units per litre were 968 in raw sewage, 12.51 in secondary effluents, 0.017 in tertiary effluents, 0.4 in river water and 0.36 in seawater. Enterovirus densities in wastewater revealed certain seasonality with a maximum at the end of spring - beginning of the summer. Coxsackievirus B, and amid them serotype CB4, were the most abundant species and serotypes detected. CONCLUSIONS: Densities of enteroviruses in different north-eastern Spain surface waters are similar to those present in industrialized countries with temperate climate. No wild polioviruses were detected. Distribution of species showed a clear prevalence of coxsackieviruses. SIGNIFICANCE AND IMPACT OF THE STUDY: Information regarding enteroviruses in this geographical area provides valuable information to estimate the risk of enteroviruses transmission through water and for complementing clinical epidemiological data. PMID- 19120642 TI - Modelling and predicting the effect of temperature, water activity and pH on growth of Streptococcus iniae in Tilapia. AB - AIMS: To evaluate and model the growth of Streptococcus iniae affect by temperatures (10-45 degrees C), water activity (A(w); 0.995-0.957), and pH (5-8). METHODS AND RESULTS: Temperatures, A(w), and pH were adjusted. The behaviour of S. iniae was studied and modelled. Growth curves were fitted by using logistic, Gompertz, and Baranyi models. The maximum growth rates obtained from the primary model were then modelled as a function of temperature, A(w), and pH using the Belehradek-type models for secondary model. The optimum values for growth were found to be in the range of 35-40 degrees C, A(w) 0.995-1, and pH 6-7. The statistical characteristics of the models were validated by r(2), mean square error, bias, and accuracy factors. The results of validation indicated that Baranyi model performed the best. CONCLUSIONS: The effect of temperature, A(w)/NaCl, pH control of S. iniae in tilapia could be satisfactorily predicted under current experimental conditions, and the proposed models could serve as a tool for this purpose. SIGNIFICANCE AND IMPACT OF THE STUDY: The suggested predictive model can be used for risk assessment concerning S. iniae in tilapia. PMID- 19120643 TI - Distribution of norovirus in oyster tissues. AB - AIMS: To study the distribution of norovirus (NV) in oyster tissues. METHODS AND RESULTS: Five monoclonal antibodies (MAbs) against VP1 were selected from Balb/c mice. Immunohistochemical analysis was performed to detect native NV in different tissues of artificially contaminated oysters using the MAbs. The data showed that the gills and the digestive glands are efficient tissues for accumulation of the NV. In addition, the NV was found on the cilia of the mantle after filtering. In our study, only NV RNA in the gills could be detected by reverse transcriptase polymerase chain reaction. CONCLUSIONS: NV was bioaccumulated in the gills, stomach, digestive diverticula and cilia of the mantle. Furthermore, the results suggested that the viral load of the gills and the digestive glands is heavier than that of the other tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: This, to our knowledge, is the first paper to report the distribution of NV in oyster tissues by immunoassay after artificial contamination. Further understanding of the NV distribution in oyster may help us to sample appropriate tissues for detection of the virus. PMID- 19120645 TI - Biosynthetic characterization and biochemical features of the third natural nisin variant, nisin Q, produced by Lactococcus lactis 61-14. AB - AIMS: To characterize the genetic and biochemical features of nisin Q. METHODS AND RESULTS: The nisin Q gene cluster was sequenced, and 11 putative orfs having 82% homology with the nisin A biosynthesis gene cluster were identified. Nisin Q production was confirmed from the nisQ-introduced nisin Z producer. In the reporter assay, nisin Q exhibited an induction level that was threefold lower than that of nisin A. Nisin Q demonstrated an antimicrobial spectrum similar to those of the other nisins. Under oxidizing conditions, nisin Q retained a higher level of activity than nisin A. This higher oxidative tolerance could be attributed to the presence of only one methionine residue in nisin Q, in contrast to other nisins that contain two. CONCLUSIONS: The 11 orfs of the nisin producers were identical with regard to their functions. The antimicrobial spectra of the three natural nisins were similar. Nisin Q demonstrated higher oxidative tolerance than nisin A. SIGNIFICANCE AND IMPACT OF THE STUDY: Genetic and biochemical features of nisin Q are similar to those of other variants. Moreover, owing to its higher oxidative tolerance, nisin Q is a potential alternative for nisin A. PMID- 19120644 TI - Genetic changes that correlate with the pine-oil disinfectant-reduced susceptibility mechanism of Staphylococcus aureus. AB - AIMS: To identify factors associated with the Staphylococcus aureus pine-oil disinfectant-reduced-susceptibility (PD(RS)) mechanism and to describe one possible PD(RS) model. METHODS AND RESULTS: Comparative genomic sequencing (CGS) and microarray analysis were utilized to detect mutations and transcriptome alterations that occur in a S. aureus PD(RS) mutant. Mutant analysis, antimicrobial gradient plates, growth studies and 3-hydroxy-3-methylglutaryl coenzyme A synthase assays were then performed to confirm the biological consequences of the 'omics' alterations detected in a PD(RS) mutant. CGS uncovered three mutations in a PD(RS) mutant in a(n): alcohol dehydrogenase (adh), catabolite control protein A (ccpA) and an NADPH-flavin oxidoreductase (frp). These mutations lead to increased growth rates; increased transcription of an NAD-dependent D-lactate dehydrogenase gene (ddh); and increased flux through the mevalonate pathway. PD(RS) mutants demonstrated reduced susceptibility to bacitracin and farnesol, and one PD(RS) mutant displayed upregulation of bacA, a bacitracin-resistance gene. Collectively, this evidence demonstrates altered undecaprenol metabolism in PD(RS) mutants. CONCLUSIONS: The PD(RS) mechanism proposed results from increased catabolic capabilities and increased flux through the mevalonate pathway as well as altered bactoprenol physiology. SIGNIFICANCE AND IMPACT OF THE STUDY: A novel mechanism that bacteria utilize to overcome the killing effects of PD formulations is proposed that is unique from the PD(RS) mechanism of the enterobacteraciae. PMID- 19120646 TI - Characterization of monoterpene biotransformation in two pseudomonads. AB - AIMS: To study the metabolic profile of Pseudomonas rhodesiae and Pseudomonas fluorescens in water-organic solvent systems using terpene substrates for both growth and biotransformation processes and to determine the aerobic or anaerobic status of these degradation pathways. MATERIALS AND METHODS: Substrates from pinene (alpha-pinene, alpha-pinene oxide, beta-pinene, beta-pinene oxide, turpentine) and limonene (limonene, limonene-1,2-oxide, orange peel oil) families were tested. For the bioconversion, the terpene-grown biomass was concentrated and used either as whole cells or as a crude enzymatic extract. CONCLUSION: Pseudomonas rhodesiae was the most suitable biocatalyst for the production of isonovalal from alpha-pinene oxide and did not metabolize limonene. Pseudomonas fluorescens was a more versatile micro-organism and metabolized limonene in two different ways. The first (anaerobic, cofactor-independent, noninducible) allowed limonene elimination by synthesizing alpha-terpineol. The second (aerobic, cofactor-dependent) involved limonene-1,2-oxide as an intermediate for energy production through a beta-oxidation process. SIGNIFICANCE AND IMPACT OF THE STUDY: Enzymatic isomerization of beta- to alpha-pinene was described for the first time for both strains. Alpha-terpineol production by P. fluorescens was very efficient and appeared as a promising alternative for the commercial production of this bioflavour. PMID- 19120647 TI - A Bayesian modelling framework to estimate Campylobacter prevalence and culture methods sensitivity: application to a chicken meat survey in Belgium. AB - AIMS: To estimate the true prevalence of Campylobacter and the diagnostic sensitivity of routine detection methods by applying a Bayesian modelling approach. METHODS AND RESULTS: Results from a Belgium-wide survey of Campylobacter contamination in chicken meat preparations (n = 656 samples) showed that Campylobacter was detected in 24.2% of the samples by enrichment, compared with 41% detected by direct plating. Combining positive results from both methods increased the apparent prevalence to 48.02%. Bayesian model was set up in WinBUGS software, the model estimates Campylobacter prevalence as 60% (95% Credibility interval (CI): 47-82%), and the sensitivity of enrichment culture and direct plating as 41% (95% CI: 31-52%) and 69% (95% CI: 50-85%), respectively. CONCLUSIONS: The parallel use of direct plating and enrichment culture adds value for Campylobacter detection from chicken meat preparations, but the false negative results from each culture method must be taken into account. SIGNIFICANCE AND IMPACT OF THE STUDY: Monitoring data could be strongly biased by the microbiological techniques used to generate it. To circumvent this bias, we describe an applied Bayesian framework for better interpretation of Campylobacter survey data in view of the imperfect test characteristics of routine culture methods. PMID- 19120648 TI - Effect of flagella expression on adhesion of Achromobacter piechaudii to chalk surfaces. AB - AIMS: To examine flagella role and cell motility in adhesion of Achromobacter piechaudii to chalk. METHODS AND RESULTS: Transmission electron microscopy revealed that stationary cells have thicker and longer flagella than logarithmic cells. SDS-PAGE analysis showed that flagellin was more abundant in stationary cells than logarithmic ones. Sonication or inhibition of flagellin synthesis caused a 30% reduction in adhesion to chalk. Preincubation of chalk with flagella extracts reduced adhesion, by 50%. Three motility mutants were isolated. Mutants 94 and 153 were nonmotile, expressed normal levels of flagellin, have regular flagella and exhibited reduced adhesion. Mutant 208 expressed low levels of flagellin, no flagella and a spherical cell shape but with normal adhesion capacity. CONCLUSIONS: Multiple cell surface factors affect the adhesion efficiency to chalk. Flagella per se through physical interaction and through cell motility contribute to the adhesion process. The adhesion behaviour of mutant 208 suggests that cell shape can compensate for flagellar removal and motility. SIGNIFICANCE AND IMPACT OF THE STUDY: Physiological status affects bacterial cell surface properties and hence adhesion efficiency to chalk. This interaction is essential to sustain biodegradation activities and thus, remediation of contaminated chalk aquifers. PMID- 19120649 TI - Molecular monitoring of inactivation efficiencies of bacteria during pulsed electric field treatment of clinical wastewater. AB - AIMS: The applicability of an alternative wastewater disinfection concept based on the pulsed electric field (PEF) treatment is tested with molecular biology techniques using clinical wastewaters. METHODS AND RESULTS: Hospital wastewater was treated with the PEF technology. The inactivation efficiencies of bacteria were successfully monitored with real-time polymerase chain reaction (PCR). As the differentiation between living and dead bacterial cells is important for the determination of the disinfection efficiency, propidium monoazide (PMA) was applied. PMA selectively penetrates cells with compromised membranes and intercalates into the DNA inhibiting a subsequent PCR amplification. The rates of reduction were examined for specific pathogens and wastewater populations using PCR-denaturing gradient gel electrophoresis. The results showed that the main part of the bacterial population could be inactivated efficiently with the PEF treatment. Moreover, it was demonstrated that naturally occurring nuclease activities were not affected by the PEF treatment in contrast to a thermal treatment. CONCLUSIONS: The results indicated that the PEF treatment is an appropriate alternative disinfection concept for the treatment of clinical wastewaters and surpass the disadvantages of other disinfection methods. SIGNIFICANCE AND IMPACT OF THE STUDY: With the use of propidium monoazide for live-dead distinction, a new concept could be developed for the evaluation of disinfection methods. PMID- 19120650 TI - Proposed mechanism of inactivating Escherichia coli O157:H7 by ultra-high pressure in combination with tert-butylhydroquinone. AB - AIMS: Investigating mechanisms of lethality enhancement when Escherichia coli O157:H7, and selected E. coli mutants, were exposed to tert-butylhydroquinone (TBHQ) during ultra-high pressure (UHP) treatment. METHODS AND RESULTS: Escherichia coli O157:H7 EDL-933, and 14 E. coli K12 strains with mutations in selected genes, were treated with dimethyl sulfoxide solution of TBHQ (15-30 ppm), and processed with UHP (400 MPa, 23 +/- 2 degrees C for 5 min). Treatment of wild-type E. coli strains with UHP alone inactivated 2.4-3.7 log CFU ml(-1), whereas presence of TBHQ increased UHP lethality by 1.1-6.2 log CFU ml(-1); TBHQ without pressure was minimally lethal (0-0.6 log reduction). Response of E. coli K12 mutants to these treatments suggests that iron-sulfur cluster-containing proteins ([Fe-S]-proteins), particularly those related to the sulfur mobilization (SUF system), nitrate metabolism, and intracellular redox potential, are critical to the UHP-TBHQ synergy against E. coli. Mutations in genes maintaining redox homeostasis and anaerobic metabolism were associated with UHP-TBHQ resistance. CONCLUSIONS: The redox cycling activity of cellular [Fe-S]-proteins may oxidize TBHQ, potentially leading to the generation of bactericidal reactive oxygen species. SIGNIFICANCE AND IMPACT OF THE STUDY: A mechanism is proposed for the enhanced lethality of UHP by TBHQ against E. coli O157:H7. The results may benefit food processors using UHP-based preservation, and biologists interested in piezophilic micro-organisms. PMID- 19120651 TI - A novel finding that Streptomyces clavuligerus can produce the antibiotic clavulanic acid using olive oil as a sole carbon source. AB - AIMS: This study aims to establish whether commercially available food oils can be used by Streptomyces clavuligerus as sole carbon sources for growth and clavulanic acid production. METHODS AND RESULTS: Batch cultures in bioreactors showed that Strep. clavuligerus growth and clavulanic acid yields in a P-limited medium containing 0.6% (v/v) olive oil were respectively 2.5- and 2.6-fold higher than in a glycerol-containing medium used as control. Glycerol- and olive oil grown cells present different macromolecular composition, particularly lipid and protein content. CONCLUSIONS: Streptomyces clavuligerus uses olive oil as the sole carbon and energy source for growth and clavulanic acid production. Yields and production rates in olive oil are comparable to those reported for oil containing complex media. Differences in yields and in the macromolecular composition indicate that different metabolic pathways convert substrate into product. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of oils being used as the sole carbon source by Strep. clavuligerus. Apart from economic benefits, interesting questions are raised about Strep. clavuligerus physiology. Defined culture media allow physiological studies to be performed in the absence of interference by other compounds. Understanding how Strep. clavuligerus catabolises oils may have an economic impact in clavulanic acid production. PMID- 19120652 TI - Laccase isoforms with unusual properties from the basidiomycete Steccherinum ochraceum strain 1833. AB - AIMS: To isolate and characterize the laccase isoforms from S. ochraceum 1833 - a new active producer of high extracellular laccase activity. METHODS AND RESULTS: Three laccase isoforms (laccases I, II and III) with 57.5, 59.5 and 63 kDa molecular masses respectively were purified from S. ochraceum 1833 and in contrast to the known laccases had strongly pronounced absorption at 611 nm with molar extinction coefficients ranging from 7170 to 7830 mol(-1) l cm(-1). All isoforms showed maximal activity with ABTS at low pH ( 0.05) however, the initial rate of germination was significantly increased at 37 degrees C compared to 20 degrees C (P < 0.05). The optimum pH range for germination was 6.5-7.5, with a decreased rate and extent of germination occurring at pH 5.5 and 8.5. CONCLUSIONS: This study demonstrates that sodium taurocholate and chenodeoxycholate initiate germination of C. difficile spores and is concentration dependant. Temperature and pH influence the rate and extent of germination. SIGNIFICANCE AND IMPACT OF THE STUDY: This manuscript enhances the knowledge of the factors influencing the germination of C. difficile spores. This may be applied to the development of potential novel strategies for the prevention of C. difficile infection. PMID- 19120668 TI - Analysis of non-Saccharomyces yeast populations isolated from grape musts from Sicily (Italy). AB - AIMS: The aim of this study was to identify the non-Saccharomyces yeast populations present in the grape must microflora from wineries from different areas around the island of Sicily. METHODS AND RESULTS: Yeasts identification was conducted on 2575 colonies isolated from six musts, characterized using Wallerstein Laboratory (WL) nutrient agar, restriction analysis of the amplified 5.8S-internal transcribed spacer region and restriction profiles of amplified 26S rDNA. In those colonies, we identified 11 different yeast species originating from wine musts from two different geographical areas of the island of Sicily. CONCLUSIONS: We isolated non-Saccharomyces yeasts and described the microflora in grape musts from different areas of Sicily. Moreover, we discovered two new colony morphologies for yeasts on WL agar never previously described. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation is a first step in understanding the distribution of non-Saccharomyces yeasts in grape musts from Sicily. The contribution is important as a tool for monitoring the microflora in grape musts and for establishing a new non-Saccharomyces yeast collection; in the future, this collection will be used for understanding the significance of these yeasts in oenology. PMID- 19120670 TI - The journal of cardiac surgery: a home where the heart is. PMID- 19120671 TI - Superior flow pattern of internal thoracic artery over saphenous vein grafts during OPCAB procedures. AB - BACKGROUND: The advantages of the use of the internal thoracic artery (ITA) over that of the saphenous vein (SV) for revascularization of the circumflex (Cx) or right coronary artery (RCA) are still debated. METHODS: Serial intraoperative flow measurements were carried out in 306 consecutive patients (mean age 64 years, mean Euroscore 5.1) undergoing off-pump coronary artery bypass grafting (OPCAB). The LAD was grafted in 302 patients [293 ITA (97%), 9 SV], the Cx in 252 patients [117 ITA (46%), 135 SV], and the RCA in 260 patients [36 ITA (14%), 224 SV]. RESULTS: Averages of 3.7+/-1.0 distal anastomoses/patient were constructed. Mean pulsatile index (PI) was significantly better for the single ITA/Cx-grafts (2.8+/-1.9, n=92) than for the single SV/Cx-grafts (3.3+/-1.7, n=43, p<0.05), whereas the mean flow did not differ (28+/-22 and 31+/-25 mL/min respectively, p=ns). Accordingly, the mean PI was significantly better for the single ITA/RCA grafts (2.2+/-1.2, n=36) than for the single SV/RCA-grafts (3.4+/-2.6, n=178, p<0.01), whereas the mean flow did not differ (30+/-16 and 32+/-22 mL/min respectively, p=ns). The incidence of perioperative myocardial infarction tended to be lower in patients receiving an ITA to either the Cx or the RCA than in those receiving a SV, but the difference did not reach statistical significance [2/92 of ITA/Cx (2.2%) vs. 2/43 of SV/Cx (4.6%), 1/36 of ITA/RCA (2.8%) vs. 8/178 of SV/RCA (4.5%)]. CONCLUSION: The internal thoracic artery provides superior flow properties than the SV to the Cx and RCA regions with reduced perioperative ischemia. Whether this advantage persists after adjusting for the grade of the proximal coronary stenosis needs further studies. PMID- 19120672 TI - Versatility of hemisternotomy for cardiac surgery. AB - BACKGROUND: Hemisternotomy has been suggested as a way to reduce morbidity by limiting the invasiveness of surgical interventions but it is often limited to aortic valve disease. This study reviews the experience of one center employing hemisternotomy and compares patient outcomes, both in-hospital and post discharge, with a matched group of full sternotomy patients. METHODS: Propensity scores were used to match all hemisternotomy valve cases (Hemi) to full sternotomy valve cases (Full) (1:2). An in-hospital composite outcome (COMP) was defined as mortality, stroke, deep sternal wound infection, sepsis, or return to operating room (OR) for bleeding or valve dysfunction. Provincial administrative health databases were used to determine freedom from mortality and hospital readmission for cardiac cause. RESULTS: During the study period, 70 patients received hemisternotomy for various cardiac surgical interventions with only 38 patients undergoing isolated aortic valve replacement. Examining valve surgery exclusively, 65 Hemi were matched to 130 Full. In-hospital complications were low in both groups, with 1.0% mortality and a non-significant trend toward COMP in the Full group (Hemi=4.6%; Full=8.5%; p=0.39). Ventilation time was significantly decreased in Hemi (median four vs. six hours; p=0.002). At two years follow-up, survival was excellent for both (Hemi=95.0%; Full=93.6%) and freedom from cardiac morbidity (Hemi=76.8%, Full=73.2%) was comparable. CONCLUSION: Hemisternotomy appears to be a safe, effective, and versatile alternative for many cardiac surgical interventions. With a median follow-up of four years, this study represents the longest cardiac morbidity follow-up for hemisternotomy patients. However, we were unable to conclusively show a morbidity benefit with this incision. PMID- 19120673 TI - In-hospital outcomes of delayed sternal closure after open cardiac surgery. AB - BACKGROUND: The purpose of this prospective study was to evaluate the current incidence, survival, morbidity, and predictors of mortality for open sternotomy at our center. METHODS: Prolonged open chest was used in 126 of 2485 cardiac operations (5.0%) between June 2006 and January 2008. The indications were hemodynamic instability (98), bleeding (14), arrhythmia (five), cardiac edema (three), tamponade (three), and cardiac arrest (three). Delayed sternal closure (DSC) was performed in 81 of 126 patients at a mean of 2.35+/-1.73 days (range: 0.5 to 10 days). RESULTS: Sixty-five of the 81 who had DSC (81.4%) survived. Forty-five patients (35.7%) died before DSC and 16 after this method. The 65 survivors (51.6%) were discharged from the hospital at a mean of 15.6+/-8.4 days (range: 7 to 50 days). The most common causes of death included low cardiac output (67.2%) and multiorgan failure (26.2%). Superficial sternal wound infection occurred in one of 81 (1.2%) patients after DSC, mediastinitis in four (4.9%), and sternal dehiscence in two (2.4%) patients. New onset of acute renal failure and the presence of intraaortic balloon pump (IABP) were predictive of in hospital death. CONCLUSIONS: This study shows that open chest with DSC is useful in the treatment of postoperatively impaired cardiac function, intractable bleeding, and persistent arrhythmias. It can be carried out with a relatively low incidence of sternal complications. Patients with new onset acute renal failure and who require IABP continue to have a high mortality rate. PMID- 19120674 TI - The Freestyle stentless bioprosthesis in more than 1000 patients: a single-center experience over 10 years. AB - BACKGROUND AND AIM: Early and mid-term clinical outcomes after aortic valve replacement (AVR) with stentless bioprostheses in a large cohort of patients are presented. METHODS: Between April 1996 and November 2005, 1014 patients underwent AVR with the stentless Medtronic Freestyle bioprosthesis, with 168 using the full root technique. The mean age was 73+/-3 (range: 20 to 90) years. Follow-up included 2953 patient-years and was 95% complete for adverse events. RESULTS: Operative mortality was 3.4% (N=34). Overall survival was 46+/-9% at nine years and similar to age- and gender-matched German general population. Freedom from prosthetic valve endocarditis, major bleeding, neurological events, and reoperation after nine years was 97+/-6%, 92+/-7%, 70+/-16%, and 92+/-9%, respectively. Freedom from structural valve deterioration was 97+/-5% at 9 years. During the learning phase, mean transprosthetic gradients of 23.5+/-3.0 mmHg and 24.8+/-3.1 mmHg were observed for valve sizes 21 and 23 mm, respectively, 10 days after subcoronary implantation in 1997, which could be lowered to 16+/-2.1 mmHg and 14.9+/-0.9 mmHg in 2005, respectively, with increasing experience of the surgeons. During the follow-up period, mean gradients dropped on average by 15 mmHg in patients presenting higher gradients at discharge. CONCLUSIONS: The Freestyle stentless bioprosthesis showed encouraging midterm durability with low rates of valve-related morbidity, and can be safely implanted without increased operative risk even during the learning phase. Special training of the surgeons is recommended to achieve optimal hemodynamic performance. PMID- 19120675 TI - Anomalous origin of the left coronary artery from the right pulmonary artery. AB - Anomalous origin of the left coronary artery (LCA) from the pulmonary artery (ALCAPA) is a rare congenital anomaly, one which requires a high index of suspicion in order to be diagnosed promptly and managed accurately. ALCAPA from the right branch pulmonary artery (RPA) is a small subset of ALCAPA in general, with only a few reported cases in the world literature. We report two cases of ALCAPA from the RPA: the first case, an infant, presented in severe cardiac failure requiring preoperative extracorporeal membrane oxygenation (ECMO) for stabilization; and the second, a neonate, presented with severe aortic coarctation (CoA). Both patients underwent successful complete surgical repair using a modified surgical technique, and are doing well in midterm follow-up. We describe our operative technique and review the literature of this rare congenital anomaly. PMID- 19120676 TI - Left internal mammary artery (LIMA) flow reserve in ischemic hypertrophied hearts. AB - It is widely accepted that in coronary artery surgery, the left internal mammary artery (LIMA) can be used as the single source of blood inflow to multiple coronary grafts as in the T-graft technique. We, herein, present clinical evidence suggesting that in ischemic hypertrophied hearts, single arterial inflow from the LIMA to multiple grafts based on the LIMA may not be sufficient and may not meet myocardial demands, at least during the early perioperative period. This observation was made in two patients in whom a vein graft, previously based on the LIMA, was also connected to the aorta. By providing additional inflow from the aorta, flows to the LAD significantly increased. PMID- 19120677 TI - The safest method of sternal wire extraction post-midsternotomy closure. AB - BACKGROUND: An incident of fatal pericardial bleeding immediately after the extraction of a sternal wire prompted a search for the most appropriate method for removing sternotomy wire sutures. A model sternum was devised to explore this problem, and several commonly used techniques for wire extraction were evaluated. METHODS: A wooden sternal model was constructed to simulate the dimensional properties of a sternum overlying the mediastinal cavity, and to imitate its tensile characteristics. A Monofil CrNi-316L (Johnson & Johnson, Brunswick, NJ, USA), No. 7 CCS, 9 metric, 4x45-cm wire was passed vertically through drilled holes. The suture was then crossed and pulled, thus joining the two boards and approximating the wire to their deep surface. A latex balloon filled with dye was placed inside under the boards. Wire holders were used to extract the wires, using a linear pulling technique and a coiling around the wire-holder tip technique. Sixty repetitions were performed for each method. RESULTS: In 60 trials of direct linear wire pulling, balloon rupture occurred in 33 (55%), whereas tearing was noted only 15 times out of 60 attempts (25%) when the tense coiling method was used. CONCLUSIONS: Sternotomy wire sutures should be extracted using a controlled technique that ensures safety to vital tissues in close proximity to the sternal bone. The tense coiling procedure offers superior safety when compared to the direct pulling process, demonstrated by a lower incidence of balloon rupture because of the lesser degree of wire flexure. This technique has become the method of choice in our medical center. PMID- 19120678 TI - Aortoentric fistula following ruptured abdominal aortic aneurysm surgery: aortobifemoral dacron bypass graft is detected by colonoscopy. AB - Aortoenteric fistula is a dramatic and rare complication, which may occur following abdominal aortic surgery. In this report, we present images of a 65 year-old patient who presented with acute peritonitis two months after aortobifemoral bypass grafting operation with a bifurcated dacron graft. Colonoscopy clearly showed the pathology leading to acute abdomen: Intestinal fistulization of the graft. PMID- 19120679 TI - Tricuspid valve myxoma: a case report and a collective review of the literature. AB - Tricuspid valve myxoma is extremely rare. An asymptomatic young patient discovered by chance an echogenic mass on the tricuspid valve by echocardiography, which was proved to be a myxoma. He underwent a successful myxoma resection via port-access approach. He had an uneventful postoperative course. Port-access approach is an alternative to conventional open heart surgical technique in terms of its minimally invasive purpose for myxoma resection. Moreover, it is of great importance in reducing the risk of intraoperative embolization and tumor implantation by minimizing the heart maneuver. PMID- 19120680 TI - Impact of minimal access valve surgery on clinical outcomes: current best available evidence. AB - Recent years have seen a surge in the enthusiasm to perform minimal access valve surgery to reduce morbidity and improve clinical outcomes. Despite tremendous enthusiasm on the part of proponents of minimal access valve surgery, skepticism still exists about the actual impact of minimal access valve surgery in reducing postoperative morbidity. This review article attempts to evaluate the current best available evidence on the impact of minimal access valve surgery on postoperative clinical outcomes. METHODS: The English language scientific literature was reviewed primarily by searching Medline from 1966 through February 2008 using PubMed interface. All blinded or unblinded randomized clinical trials, comparing minimal access valve surgery with conventional valve surgery through a full sternotomy, recruiting adult human patients undergoing valve repair, or replacement and reporting impact of these two approaches on at least 1 pertinent clinical or economic outcome, were included. RESULTS: Current best available evidence from randomized clinical trials (Grade A, Level 1b) does not show any significant quantitative differences between minimal access valve surgery and conventional valve surgery for perioperative mortality or other primary outcome events of stroke, renal failure, or respiratory failure. There are small but statistically significant benefits for minimal access valve surgery for surrogate outcomes of ventilation time, intensive care unit stay, and total length of stay. CONCLUSION: The published evidence is thin and a large multicenter randomized clinical trial with preferably standardization of minimal access valve surgery techniques and long term follow-up is required to validate the safety and efficacy of minimal access valve surgery. PMID- 19120681 TI - Mitral valve replacement in a patient with idiopathic hypereosinophilic syndrome and pulmonary arterial hypertension. AB - BACKGROUND AND AIMS: Idiopathic hypereosinophilic syndrome, a rarely seen systemic disease, may cause cardiac valvular lesions by eosinophilic infiltration. This report describes management of a 25-year-old woman with idiopathic hypereosinophilic syndrome, severe mitral stenosis, and pulmonary arterial hypertension. METHODS: The patient was presented with haemoptysia and dyspnea on exertion. Echocardiography showed severe mitral stenosis and pulmonary arterial hypertension. RESULTS: After hematological stabilization, she underwent mitral valve replacement using a No. 27 bovine pericardial valve. In the intensive care unit she had a pulmonary hypertensive crisis, which ameliorated gradually with sedation and nitroglycerin. She was extubated and discharged on the second and seventh days, respectively. CONCLUSION: Surgical experience for the patients with mitral dysfunction caused by idiopathic hypereosinophilic syndrome is limited. When mitral valve replacement is needed, the ideal type of prosthesis remains unclear and the presence of pulmonary arterial hypertension further complicates the management. We think that bioprosthetic valves would be the appropriate choice in eosinophilic mitral dysfunction requiring valve replacement. PMID- 19120682 TI - Left ventricular free wall rupture subsequent to early myocardial reinfarction after successful percutaneous transluminal coronary angiography. AB - Left ventricular free wall rupture is a rare complication of acute myocardial infarction. Scarcely has a cardiac rupture subsequent to early reinfarction been reported. A left ventricular free wall rupture subsequent to reinfarction after successful percutaneous transluminal coronary angiography in a 78-year-old female is herein described. PMID- 19120683 TI - Ventricular fibrillation with prominent early repolarization associated with a rare variant of KCNJ8/KATP channel. AB - BACKGROUND: Early repolarization in the inferolateral leads has been recently recognized as a frequent syndrome associated with idiopathic ventricular fibrillation (VF). We report the case of a patient presenting dramatic changes in the ECG in association with recurrent VF in whom a novel genetic variant has been identified. CASE REPORT: This young female (14 years) was resuscitated in 2001 following an episode of sudden death due to VF. All examinations including coronary angiogram with ergonovine injection, MRI, and flecainide or isoproterenol infusion were normal. The patient had multiple (>100) recurrences of VF unresponsive to beta-blockers, lidocaine/mexiletine, verapamil, and amiodarone. Recurrences of VF were associated with massive accentuation of the early repolarization pattern at times mimicking acute myocardial ischemia. Coronary angiography during an episode with 1.2 mV J/ST elevation was normal. Isoproterenol infusion acutely suppressed electrical storms, while quinidine eliminated all recurrences of VF and restored a normal ECG over a follow-up of 65 months. Genomic DNA sequencing of K(ATP) channel genes showed missense variant in exon 3 (NC_000012) of the KCNJ8 gene, a subunit of the K(ATP) channel, conferring predisposition to dramatic repolarization changes and ventricular vulnerability. PMID- 19120684 TI - Re: Beta1 and beta2-adrenergic receptor polymorphisms and idiopathic ventricular arrhythmias. PMID- 19120686 TI - Fractionated electrograms in atrial fibrillation. PMID- 19120688 TI - Isolation and characterization of DUSP11, a novel p53 target gene. AB - p53 regulates the expression of genes involved in cell cycle control, apoptosis and DNA damage repair. Here we demonstrate that DUSP11 (dual specificity phosphatase 11), a member of the protein tyrosine phosphatase family that binds to RNA-RNP complexes and RNA splicing factors, is a p53 target gene. Consistent with this, the expression of DUSP11 is induced in a p53-dependent manner after treatment with DNA damaging agents. Chromatin immunoprecipitation analysis showed that p53 binds to 2 putative p53 DNA binding sites in the promoter region of DUSP11. Colony formation and proliferation assays demonstrated that the ectopic expression of wildtype, but not catalytical inactive, DUSP11 leads to growth arrest. Furthermore inhibition of DUSP11 expression by shRNA increases the proliferation of normal and DNA damaged cells in tissue culture. Finally we show that the splicing factor SAM68 (Src-associated protein in mitotic cells) binds to DUSP11 in vitro and in vivo. Taken together these results suggest that DUSP11 contributes to p53-dependent inhibition of cell proliferation and that it might be involved in regulating RNA splicing through SAM68. PMID- 19120689 TI - Up-regulation of reverse cholesterol transport key players and rescue from global inflammation by ApoA-I(Milano). AB - Recombinant-ApoA-I(M) (rApoA-I(M)) administration has been shown to regress and stabilize atherosclerotic plaques. However, the mechanisms responsible for these beneficial effects are not fully understood. The aims of the present study were to define whether the benefits of rApoA-I(M) treatment were mediated via an enhanced reverse cholesterol transport (RCT) and/or anti-inflammation-related mechanisms. Advanced aortic lesions were induced in New Zealand White rabbits (n= 16). Animals were randomized to placebo or rApoA-I(M) (rApoA-I(M)/phospholipids; ETC-216), two infusions 4 days apart. Four days after last dose, aortas and livers were processed for cholesterol content, expression of RCT-related receptors (ATP-binding cassette A-1 [ABCA-1] and scavenger receptor BI [SR-BI]), and inflammation-related markers (inducible nitric oxide synthase [iNOS] and capase-3). Oxidative stress was assessed in the vessel wall and in plasma. rApoA I(M) administration resulted in a significant reduction in the hepatic and aortic cholesterol content without differences in plasma levels. This effect was associated with an up-regulation of vessel wall ABCA-1, as well as a hepatic and arterial-wall SR-BI up-regulation. Systemic and atherosclerotic-plaque inflammation markers were significantly reduced by the rApoA-I(M) administration, as demonstrated by a reduction in circulating oxidative stress markers and prostaglandin F1-alpha levels, and the down-regulation of the iNOS and caspase 3 in the aortic lesions. rApoA-IM up-regulated the ABCA-1 and SR-BI levels to a greater extent than the wild-type form of apoA-I in in vitro studies done with lipid-rich macrophages. Our data suggest that rApoA-I(M) administration enhances RCT and induces a 'rescue' from the global inflammatory status associated with atherosclerotic disease. The Milano form of apoA-I seems to be more efficient in RCT than the apoA-I wild-type. PMID- 19120690 TI - Antagomir-mediated silencing of endothelial cell specific microRNA-126 impairs ischemia-induced angiogenesis. AB - MicroRNAs are negative regulators of gene expression that play a key role in cell type specific differentiation and modulation of cell function and have been proposed to be involved in neovascularization. Previously, using an extensive cloning and sequencing approach, we identified miR-126 to be specifically and highly expressed in human endothelial cells (EC). Here, we demonstrate EC specific expression of miR-126 in capillaries and the larger vessels in vivo. We therefore explored the potential role of miR-126 in arteriogenesis and angiogenesis. Using miR-reporter constructs, we show that miR-126 is functionally active in EC in vitro and that it could be specifically repressed using antagomirs specifically targeting miR-126. To study the consequences of miR-126 silencing on vascular regeneration, mice were injected with a single dose of antagomir-126 or a control 'scramblemir' and exposed to ischemia of the left hindlimb by ligation of the femoral artery. Although miR-126 was effectively silenced in mice treated with a single, high dose (HD) of antagomir-126, laser Doppler perfusion imaging did not show effects on blood flow recovery. In contrast, quantification of the capillary density in the gastrocnemius muscle revealed that mice treated with a HD of antagomir-126 had a markedly reduced angiogenic response. Aortic explant cultures of the mice confirmed the role of miR-126 in angiogenesis. Our data demonstrate a facilitary function for miR-126 in ischemia-induced angiogenesis and show the efficacy and specificity of antagomir-induced silencing of EC-specific microRNAs in vivo. PMID- 19120691 TI - Measuring and monitoring apoptosis and drug toxicity in HIV patients by ligation mediated polymerase chain reaction. AB - Apoptosis has a critical role in normal physiology while its dysregulation has causal links with certain pathologies. A biochemical hallmark of apoptosis, internucleosomal genomic DNA fragmentation, is detectable by ligation-mediated polymerase chain reaction (LM-PCR). Here we converted LM-PCR into a new apoptosis quantifier by dividing trace quantities of 600 bp apoptotic amplicons into those of a single copy house-keeping gene, generating the LM-PCR 'value'. Dynamic range was approximately 17-fold correlating with a approximately 200-fold difference in degree of apoptotic fragmentation. Inter- and intra-gel reliability were both excellent, supporting LM-PCR's utility with large sample sets. Validation experiments comprising cell exposure to staurosporine over time revealed LM-PCR is as sensitive as caspase-3/ELISA and more sensitive than terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling/flourescence activated cell sorting (TUNEL/FACS) for distinguishing low degrees of apoptosis (the spectrum most relevant in vivo). The LM-PCR profile mirrored that of caspase 3/ELISA but not TUNEL/FACS. We then applied this molecular tool to clinical investigation. Increased apoptosis is implicated in lipoatrophy (subcutaneous fat wasting), a serious, persistent toxicity of some nucleoside analogue reverse transcriptase inhibitors (NRTIs) used in anti-HIV highly active antiretroviral therapy (HAART). We demonstrated in 105 peripheral blood mononuclear cell samples that elevated LM-PCR values are seen during therapy with stavudine (d4T), a particularly toxic NRTI (P< 0.0001 versus no HAART, unpaired t-test). Elevated values were also independently associated with clinical evidence of lipoatrophy (P= 0.007, multiple logistic regression modelling) but not with patient age, CD4 T-cell count nor HIV viral load (P> 0.8 for each). Together these data demonstrate that LM-PCR is a robust and reliable quantifier of apoptosis with potential for basic science and clinical investigation. PMID- 19120692 TI - Haemopexin affects iron distribution and ferritin expression in mouse brain. AB - Haemopexin (Hx) is an acute phase plasma glycoprotein, mainly produced by the liver and released into plasma where it binds heme with high affinity and delivers it to the liver. This system provides protection against free heme mediated oxidative stress, limits access by pathogens to heme and contributes to iron homeostasis by recycling heme iron. Hx protein has been found in the sciatic nerve, skeletal muscle, retina, brain and cerebrospinal fluid (CSF). Recently, a comparative proteomic analysis has shown an increase of Hx in CSF from patients with Alzheimer's disease, thus suggesting its involvement in heme detoxification in brain. Here, we report that Hx is synthesised in brain by the ventricular ependymal cells. To verify whether Hx is involved in heme scavenging in brain, and consequently, in the control of iron level, iron deposits and ferritin expression were analysed in cerebral regions known for iron accumulation. We show a twofold increase in the number of iron-loaded oligodendrocytes in the basal ganglia and thalamus of Hx-null mice compared to wild-type controls. Interestingly, there was no increase in H- and L-ferritin expression in these regions. This condition is common to several human neurological disorders such as Alzheimer's disease and Parkinson's disease in which iron loading is not associated with an adequate increase in ferritin expression. However, a strong reduction in the number of ferritin-positive cells was observed in the cerebral cortex of Hx-null animals. Consistent with increased iron deposits and inadequate ferritin expression, malondialdehyde level and Cu-Zn superoxide dismutase-1 expression were higher in the brain of Hx-null mice than in that of wild-type controls. These data demonstrate that Hx plays an important role in controlling iron distribution within brain, thus suggesting its involvement in iron-related neurodegenerative diseases. PMID- 19120695 TI - Small-molecule peptides inhibit Z alpha1-antitrypsin polymerization. AB - The Z variant of 1-antitrypsin (AT) polymerizes within the liver and gives rise to liver cirrhosis and the associated plasma deficiency leads to emphysema. In this work, a combinatorial approach based on the inhibitory mechanism of (alpha1) AT was developed to arrest its pathogenic polymerization. One peptide, Ac-TTAI NH(2), emerged as the most tight-binding ligand for Z (alpha1)-AT. Characterization of this tetrapeptide by gel electrophoresis and biosensor analysis revealed its markedly improved binding specificity and affinity compared with all previously reported peptide inhibitors. In addition, the peptide is not cytotoxic to lung cell lines. A model of the peptide-protein complex suggests that the peptide interacts with nearby residues by hydrogen bonds, hydrophobic interactions, and cavity-filling stabilization. The combinatorially selected peptide not only effectively blocks the polymerization but also promotes dissociation of the oligomerized (alpha1)-AT. These results are a significant step towards the potential treatment of Z (alpha1)-AT related diseases. PMID- 19120693 TI - Dexamethasone inhibits lipopolysaccharide-induced hydrogen sulphide biosynthesis in intact cells and in an animal model of endotoxic shock. AB - Dexamethasone (1 mg/kg, i.p.) administered either 1 hr before or 1 hr after E. coli lipopolysaccharide (LPS, 4 mg/kg, i.p.) in conscious rats inhibited the subsequent (4 hrs) rise in plasma cytokine (interleukin [IL]-1beta, tumour necrosis factor [TNF]-alpha), nitrate/nitrite (NO(x)), soluble intercellular adhesion molecule-1 (sICAM-1) concentration and lung/liver myeloperoxidase activity indicative of an anti-inflammatory effect. Dexamethasone also reduced the LPS-evoked rise in plasma hydrogen sulphide (H(2)S) concentration, liver H(2)S synthesizing activity and expression of cystathionine gamma lyase (CSE) and inducible nitric oxide synthase (iNOS). Mifepristone (RU-486) inhibited these effects. Dexamethasone (1-10 microM) reduced the LPS-evoked release of IL-1beta, TNF-alpha and L-selectin, decreased expression of CSE and iNOS and diminished nuclear factor kappaB (NF-kappaB)-DNA binding in isolated rat neutrophils. In contrast, NaHS (100 microM) increased L-selectin release from rat neutrophils. Dexamethasone also reduced LPS-induced up-regulation of CSE in foetal liver cells. 6-amino-4-(4-phenoxyphenylethylamino) quinazoline (QNZ, 10 nM), a selective inhibitor of transcription via the NF-kappaB pathway, abolished LPS induced up-regulation of CSE expression. We propose that inhibition of CSE expression and reduction in formation of the pro-inflammatory component of H(2)S activity contributes to the anti-inflammatory effect of dexamethasone in endotoxic shock. Whether H(2)S plays a part in the anti-inflammatory effect of this steroid in other forms of inflammation such as arthritis or asthma warrants further study. PMID- 19120697 TI - The role of CCAAT/enhancer binding protein (C/EBP)-alpha in osteogenesis of C3H10T1/2 cells induced by BMP-2. AB - The balance between osteogenesis and adipogenesis of mesenchymal stem cells is disrupted in various human diseases. Investigating the mechanisms that fine-tune this balance is of medical importance. Identification of potential target gene which can be used to study the relationship between them could be really helpful for this purpose. In the current study, we used C3H10T1/2 as model cells and through which two models of both osteogenesis induced by bone-morphogenetic protein (BMP)-2 and transdifferentiation from osteogenesis to adipogenesis were established. We investigated the role of CCAAT/enhancer binding protein (C/EBP) alpha in these two systems. Then from epigenetic point of view, we elucidated the underlying molecular mechanisms preliminarily. The results showed that down regulations of both C/EBP-alpha expression and its inducibility in response to insulin, fetal bovine serum, methylisobutylxanthine and dexamethasone (IFMD) adipogenic hormonal cocktail were observed in terminal stage of osteogenesis of C3H10T1/2 cells induced by BMP-2. And overexpression of C/EBP-alpha could lead to inhibition of osteogenesis differentiation and rescue attenuation of potential of adipogenic conversion in this process. Furthermore, we provided evidence that remarkable DNA hypermethylation and histones 3 and 4 hypoacetylation in -1286 bp/1065 bp promoter region of C/EBP-alpha were involved in both of down regulations. Our data suggest that C/EBP-alpha functions as regulator in the balance between osteogenesis and adipogenesis of C3H10T1/2 cells and may be a therapeutic target. PMID- 19120699 TI - Contractility in health and disease. PMID- 19120698 TI - Structural and mechanistic insights into Mps1 kinase activation. AB - Mps1 is one of the several essential kinases whose activation is required for robust mitotic spindle checkpoint signalling. The activity of Mps1 is tightly regulated and increases dramatically during mitosis or in response to spindle damage. To understand the molecular mechanism underlying Mps1 regulation, we determined the crystal structure of the kinase domain of Mps1. The 2.7-A resolution crystal structure shows that the Mps1 kinase domain adopts a unique inactive conformation. Intramolecular interactions between the key Glu residue in the C helix of the N-terminal lobe and the backbone amides in the catalytic loop lock the kinase in the inactive conformation. Autophosphorylation appears to be a priming event for kinase activation. We identified Mps1 autophosphorylation sites in the activation and the P+1 loops. Whereas activation loop autophosphorylation enhances kinase activity, autophosphorylation at the P+1 loop (T686) is associated with the active kinase. Mutation of T686 autophosphorylation site impairs both autophosphorylation and transphosphorylation. Furthermore, we demonstrated that phosphorylation of T676 may be a priming event for phosphorylation at T686. Finally, we identified two critical lysine residues in the loop between helices EF and F that are essential for substrate recruitment and maintaining high levels of kinase activity. Our studies reveal critical biochemical mechanisms for Mps1 kinase regulation. PMID- 19120700 TI - The potential role of MLC phosphatase and MAPK signalling in the pathogenesis of vascular dysfunction in heart failure. AB - The clinical syndrome of heart failure is associated with both a resting vasoconstriction and reduced sensitivity to nitric oxide mediated vasodilatation, and this review will focus on the role of myosin light chain (MLC) phosphatase in the pathogenesis of the vascular abnormalities of heart failure. Nitric oxide mediates vasodilatation by an activation of guanylate cyclase and an increase in the production of cGMP, which leads to the activation of the type I cGMP dependent protein kinase (PKGI). PKGI then activates a number of targets that produce smooth muscle relaxation including MLC phosphatase. MLC phosphatase is a holoenzyme consisting of three subunits; a 20 kD subunit of unknown function, an approximately 38-kD catalytic subunit and a myosin targeting subunit (MYPT1). Alternative splicing of a 31 bp 3 exon generates MYPT1 isoforms, which differ by a COOH-terminus leucine zipper (LZ). Further, PKGI-mediated activation of MLC phosphatase requires the expression of a LZ+ MYPT1. Congestive heart failure is associated with a decrease in LZ+ MYPT1 expression, which results in a decrease in the sensitivity to cGMP-mediated smooth muscle relaxation. Beyond their ability to reduce afterload, angiotensin converting enzyme (ACE) inhibitors have a number of beneficial effects that include maintaining the expression of the LZ+ MYPT1 isoform, thereby conserving normal sensitivity to cGMP-mediated vasodilatation, as well as differentially regulating genes associated with mitogen activated protein kinase (MAPK) signalling. ACE inhibition reduces circulating angiotensin II and thus limits the downstream activation of MAPK signalling pathways, possibly preventing the alteration of the vascular phenotype to preserve normal vascular function. PMID- 19120702 TI - MicroRNAs in embryonic stem cells and early embryonic development. AB - MicroRNAs (miRNAs) are involved in the regulation of a broad range of biological processes. There is an extensive literature on the roles of miRNAs in metazoan development, from early embryogenesis to lineage commitment, and formation and maturation of cellular subtypes and complex tissues. This review will present the evidence that miRNAs are essential for the earliest stages of metazoan development, including establishment and maintenance of the pluripotent embryonic stem cells (ESCs) from which all foetal tissues arise. We will also discuss specific miRNAs that have been associated with early embryogenesis and pluripotency. Finally, we will point out evidence showing that there is strong and highly redundant miRNA regulation of a common set of target genes in the early stages of embryonic development and in ESCs. PMID- 19120705 TI - Editorial: neurobiological models of childhood depression and parenting models of childhood anxiety. PMID- 19120701 TI - Smooth muscle signalling pathways in health and disease. AB - Smooth muscle contractile activity is a major regulator of function of the vascular system, respiratory system, gastrointestinal system and the genitourinary systems. Malfunction of contractility in these systems leads to a host of clinical disorders, and yet, we still have major gaps in our understanding of the molecular mechanisms by which contractility of the differentiated smooth muscle cell is regulated. This review will summarize recent advances in the molecular understanding of the regulation of smooth muscle myosin activity via phosphorylation/dephosphorylation of myosin, the regulation of the accessibility of actin to myosin via the actin-binding proteins calponin and caldesmon, and the remodelling of the actin cytoskeleton. Understanding of the molecular 'players' should identify target molecules that could point the way to novel drug discovery programs for the treatment of smooth muscle disorders such as cardiovascular disease, asthma, functional bowel disease and pre-term labour. PMID- 19120704 TI - Statins are detrimental to human placental development and function; use of statins during early pregnancy is inadvisable. PMID- 19120706 TI - Emanuel Miller Lecture: early onset depressions--meanings, mechanisms and processes. AB - BACKGROUND: Depressive syndromes in children and adolescents constitute a serious group of mental disorders with considerable risk for recurrence. A more precise understanding of aetiology is necessary to improve treatment and management. METHODS: Three neuroactive agents are purported to be involved in the aetiology of these disorders: serotonin, brain-derived neurotrophic factor and cortisol. A literature review was conducted to determine their contributions to the emergence of unipolar depressions in the adolescent years. RESULTS: Serotonin, brain derived neurotrophic factor and cortisol may operate in concert within two distinct functional frameworks: atypical early epigenesis arising in the first few years of life and resulting in the formation of a vulnerable neuronal network involving in particular the amygdala and ventral prefrontal cortex. Individuals with this vulnerability are likely to show impaired mood regulation when faced with environmental demands during adolescence and over the subsequent decades; and acquired neuroendangerment, a pathological brain process leading to reduced synaptic plasticity, in particular in the hippocampus and perhaps the nucleus accumbens and ventral tegmentum. This may result in motivational, cognitive and behavioural deficits at any point in the lifespan most apparent at times of environmental demand. CONCLUSIONS: The characteristics, course and outcome of a depressive episode may depend on the extent of the involvement of both atypical early neurogenesis and acquired neuroendangerment. PMID- 19120707 TI - Learned social hopelessness: the role of explanatory style in predicting social support during adolescence. AB - BACKGROUND: Almost no research has examined the impact of explanatory style on social adjustment. We hypothesised that adolescents with a pessimistic style would be less likely to develop and maintain social support networks. METHODS: Seven hundred and nineteen students (351 males and 366 females; 2 unknown; M(AGE) = 12.28, SD = .49) completed an anonymous survey in Grades 7 through 10. Explanatory style was assessed in Grades 7 and 9, sadness was assessed in Grades 7 through 10, and quantity and quality of social support was assessed in Grades 8, 9, and 10. RESULTS: Structural equation modelling was used to conduct cross lagged panel analyses of the four waves of data. Pessimistic explanatory style predicted lower levels of social support, and lower social support from the family predicted higher levels of pessimistic explanatory style. Additional analyses suggested that the effects could not be explained by sadness or by assuming that pessimistic adolescents where less liked by their peers. CONCLUSIONS: Pessimistic adolescents feel unable to influence their social worlds in positive ways and consequently may not take actions to develop and maintain social support networks. PMID- 19120703 TI - MicroRNA involvement in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the third cause of cancer-related death worldwide. Curative options for HCC are limited and exclusively available for patients carrying an early stage HCC. In advanced stages, traditional chemotherapy proved to be only marginally effective or even toxic. Thus, the identification of new treatment options is needed. New targets for non conventional treatment will necessarily take advantage of progresses on the molecular pathogenesis of HCC. MicroRNAs (miRNAs) are a group of tiny RNAs with a fundamental role in the regulation of gene expression. Aberrant expression of several miRNAs was found to be involved in human hepatocarcinogenesis. miRNA expression signatures were correlated with bio-pathological and clinical features of HCC. In some cases, aberrantly expressed miRNAs could be linked to cancer associated pathways, indicating a direct role in liver tumourigenesis. For example, up-regulation of mir-221 and mir-21 could promote cell cycle progression, reduce cell death and favour angiogenesis and invasion. These findings suggest that miRNAs could become novel molecular targets for HCC treatment. The demonstration of in vivo efficacy and safety of anti-miRNA compounds has opened the way to their use in clinical trials. PMID- 19120708 TI - Impaired prioritization of novel onset stimuli in autism spectrum disorder. AB - BACKGROUND: Deficiency in the adaptive allocation of attention to relevant environmental stimuli is an associated feature of autism spectrum disorder (ASD). Recent evidence suggests that individuals with ASD may be specifically impaired in attentional prioritization of novel onsets. METHOD: We investigated modulation of attention by novel onset stimuli in 22 children with ASD and 22 age- and IQ matched typically developing (TD) children using a preview visual search task (Donk & Theeuwes, 2003). In preview search, a subset of search stimuli (old) is presented briefly before the remaining stimuli (new) with the effect that search times for targets appearing among the new elements are typically shorter than for those appearing among the old elements. RESULTS: Whereas the TD group exhibited faster reaction time (RT) to targets occurring as novel search elements, the ASD group performed similarly in target new and old conditions, indicating impaired attentional prioritization of novel onsets. Group differences in eye-movement behavior, including fixation frequency and saccadic error for novel onset stimuli, were consistent with the RT findings. Attentional modulation by novel onsets varied inversely with social-communicative symptom severity in the ASD group. CONCLUSIONS: The results provide further evidence of reduced sensitivity to novel onsets in ASD, and suggest that impaired processing of dynamic stimuli, possibly associated with abnormalities in the dorsal visual processing stream, may be implicated in the core symptoms of ASD. PMID- 19120709 TI - Construct validity of the five-factor Strengths and Difficulties Questionnaire (SDQ) in pre-, early, and late adolescence. AB - BACKGROUND: The Strengths and Difficulties Questionnaire (SDQ) is designed to measure psychological adjustment in children and adolescents. Psychometric evaluations of the instrument have shown satisfactory convergent and discriminant validity, while factor analysis studies have shown mixed results across countries. In the present study, the construct validity of the five-factor SDQ is evaluated in a large community sample of Norwegian pre-, early, and late adolescents. METHODS: The sample consisted of 26,269 children and adolescents (10 19 years) with valid answers on all 25 items of the SDQ self-report. Complete parent/proxy data of respective pre-adolescent children was available for 6,645 cases. A Lisrel approach to Confirmatory Factor Analysis (CFA) was used to evaluate the five-factor model and the presence of a positive construal factor. In the sample of pre-adolescents and their parents/proxies, convergent and discriminant validity was evaluated by a CFA approach to multitrait-multimethods (MTMM). RESULTS: Fit statistics for the hypothesized five-factor model were satisfactory, but introducing correlated error terms for some of the items led to significant model improvement in all age groups. All factor loadings were higher than .30, except for item 11 (good friend). The loadings differed across age groups and differed markedly between the parent/proxy and self-report measures. The MTMM showed that the source of ratings made a difference on the validity of all subscale ratings, with self-reports discriminating more on ratings of emotional and peer problems, and parents/proxies discriminating more on hyperactivity symptoms. A positive construal factor was identified but had a modest effect compared with the original five traits. Results suggested an unclear construct and meaning of the Prosocial behaviour subscale. CONCLUSION: The results of the present study indicated support for the proposed five-factor structure of the SDQ (Goodman, 2001) across a wide age range (10-19years), including older adolescents and different informants. However, some improvements should be considered to improve internal reliability and conceptual clarity. PMID- 19120711 TI - Verbal short-term memory span in children: long-term modality dependent effects of intrauterine growth restriction. AB - BACKGROUND: Recent reports showed that children born with intrauterine growth restriction (IUGR) are at greater risk of experiencing verbal short-term memory span (STM) deficits that may impede their learning capacities at school. It is still unknown whether these deficits are modality dependent. METHODS: This long term, prospective design study examined modality-dependent verbal STM functions in children who were diagnosed at birth with IUGR (n = 138) and a control group (n = 64). Their STM skills were evaluated individually at 9 years of age with four conditions of the Visual-Aural Digit Span Test (VADS; Koppitz, 1981): auditory-oral, auditory-written, visuospatial-oral and visuospatial-written. Cognitive competence was evaluated with the short form of the Wechsler Intelligence Scales for Children--revised (WISC-R95; Wechsler, 1998). RESULTS: We found IUGR-related specific auditory-oral STM deficits (p < .036) in conjunction with two double dissociations: an auditory-visuospatial (p < .014) and an input output processing distinction (p < .014). Cognitive competence had a significant effect on all four conditions; however, the effect of IUGR on the auditory-oral condition was not overridden by the effect of intelligence quotient (IQ). CONCLUSIONS: Intrauterine growth restriction affects global competence and inter modality processing, as well as distinct auditory input processing related to verbal STM functions. The findings support a long-term relationship between prenatal aberrant head growth and auditory verbal STM deficits by the end of the first decade of life. Empirical, clinical and educational implications are presented. PMID- 19120710 TI - G x E interaction in the organization of attachment: mothers' responsiveness as a moderator of children's genotypes. AB - BACKGROUND: Scholars acknowledge that both biologically-founded child temperament and environmental factors are influential in determining the quality of infant mother attachment. We present evidence for gene by environment (G x E) interaction in the organization of attachment. METHODS: Participants were 88 typically developing infants and their mothers. Molecular genetic measures of the infants focused on the polymorphism in the serotonin transporter gene (5-HTTLPR, ss/sl vs. ll genotype). Mothers' responsiveness to their infants at 7 months was observed in lengthy naturalistic interactions, and was conceptualized as the environmental influence. RESULTS: For infants with a short allele (ss/sl ), variation in mothers' responsiveness was significantly associated with attachment security, assessed at 15 months in the Strange Situation. For those infants, low responsiveness predicted particularly high risk for insecure attachment, and high responsiveness offset that risk. For infants homozygous for the long allele (ll ), there was no association between responsiveness and attachment organization. CONCLUSIONS: The findings show that the quality of early care serves to amplify or offset the risk conferred by genotype. PMID- 19120713 TI - Impaired conflict resolution and alerting in children with ADHD: evidence from the Attention Network Task (ANT). AB - BACKGROUND: An important theory of attention suggests that there are three separate networks that execute discrete cognitive functions. The 'alerting' network acquires and maintains an alert state, the 'orienting' network selects information from sensory input and the 'conflict' network resolves conflict that arises between potential responses. This theory holds promise for dissociating discrete patterns of cognitive impairment in disorders where attentional deficits may often be subtle, such as in attention deficit hyperactivity disorder (ADHD). METHODS: The Attentional Network Test (ANT), a behavioural assay of the functional integrity of attention networks, was used to examine the performance of 73 children with ADHD and 73 controls. RESULTS: Performance on the ANT clearly differentiated the children with and without ADHD in terms of mean and standard deviation (SD) of reaction time (RT), the number of incorrect responses made and the number of omission errors made. The ADHD group demonstrated deficits in the conflict network in terms of slower RT and a higher number of incorrect responses. The ADHD group showed deficits in the alerting network in terms of the number of omission errors made. There was no demonstration of a deficit in the orienting network in ADHD on this task. CONCLUSIONS: The children with ADHD demonstrated deficits in the alerting and conflict attention networks but normal functioning of the orienting network. PMID- 19120714 TI - High blood pressure in children and adolescents: to treat or not to treat is not the question. PMID- 19120712 TI - Association of ADHD, tics, and anxiety with dopamine transporter (DAT1) genotype in autism spectrum disorder. AB - BACKGROUND: Autism spectrum disorder (ASD) is associated with high rates of psychiatric disturbance to include attention-deficit/hyperactivity disorder (ADHD), tic disorder, and anxiety disorders. The aim of the present study was to examine the association between a variable number tandem repeat (VNTR) functional polymorphism located in the 3'-untranslated region of the dopamine transporter gene (DAT1) and the severity of these symptoms as well as the association between the DAT1 DdeI polymorphism and severity of tics. METHODS: Parents (n = 62) and teachers (n = 57) completed a DSM-IV-referenced rating scale for 67 children with ASD. RESULTS: According to parent ratings, children with the 10-10 repeat allele (versus a combined group of all other genotypes) exhibited less severe symptoms of hyperactivity and impulsivity as well as less severe language deficits. Teacher ratings indicated that social anxiety and tic symptoms were more severe for children with the 10-10 genotype versus all others. Exploratory analyses provided preliminary support for the notion that heterozygosity (9-10 repeat genotype) may be a risk/protective factor. There were no associations of tic severity with the DAT1 DdeI polymorphism. CONCLUSION: Collectively, these results suggest that the extraordinary variability in ASD clinical phenotypes may be explained in part by the same genes that are implicated in a host of other psychiatric disorders in non-ASD populations. Nevertheless, replication with independent samples is necessary to confirm this preliminary finding. PMID- 19120715 TI - Metabolic and antihypertensive effects of combined angiotensin receptor blocker and diuretic therapy in prediabetic hypertensive patients with the cardiometabolic syndrome. AB - Hypertensive patients with the cardiometabolic syndrome (CMS) are at increased risk for type 2 diabetes and cardiovascular disease. The authors examined effects of valsartan and hydrochlorothiazide (HCTZ) combined and alone on insulin sensitivity (using homeostasis model assessment-insulin resistance [HOMA-IR]), and inflammatory/metabolic biomarkers in prediabetic hypertensive persons with CMS. Eligible patients entered 16-week therapy with valsartan 320 mg/d (n=189), HCTZ 25 mg/d (n=190), or valsartan/HCTZ 320/25 mg/d (n=187). At the end point, there were no statistically significant differences in HOMA-IR among the 3 groups. HCTZ significantly increased hemoglobin A(1c) and triglyceride concentrations and lowered serum potassium levels vs valsartan. HCTZ also increased plasma aldosterone and C-reactive protein levels. Blood pressure reduction and blood pressure control rates were highest with valsartan/HCTZ. There were no differences between combination valsartan/HCTZ or monotherapies on a measure of insulin sensitivity; however, the negative metabolic effects of HCTZ (increase in triglyceride and hemoglobin A(1c) values) were absent with valsartan/HCTZ, indicating an ameliorating effect of valsartan on these measures. PMID- 19120716 TI - Sex differences in short-term heart rate variability in patients with newly diagnosed essential hypertension. AB - The authors report the results of the analysis of heart rate variability (HRV) indices during 5 minutes of supine rest and 5 minutes of standing and conventional indices of autonomic function in 69 men and 51 women with untreated newly diagnosed hypertension matched for body mass index and resting blood pressure. Mean RR interval, standard deviation of normal-to-normal RR intervals, low-frequency RR spectral power, HRV during deep breathing at 6 breaths per minute, and the 30:15 ratio (maximum RR interval 30th beat/minimum RR interval 15th beat) were significantly lower in women (P=.01, .02, .001, .04, .01, respectively) compared with men. Low frequency RR in normalized units was lower in women in the supine position alone (P=.03). HRV was significantly lower in women with untreated newly diagnosed hypertension compared with men. The authors interpret these results as indicating an increase in baseline cardiac sympathovagal balance in female hypertensive patients. PMID- 19120717 TI - Results of an olmesartan medoxomil-based treatment regimen in hypertensive patients. AB - The efficacy and safety of an olmesartan medoxomil (OM)-based treatment algorithm was tested in a double-blind, randomized, placebo-controlled titration study in 276 patients with stage 1 or 2 hypertension. After placebo run-in, patients were randomized to placebo (12 weeks) or OM 20 mg/d (weeks 1-3). OM was up-titrated to 40 mg/d (weeks 4-6), then OM/hydrochlorothiazide (HCTZ) 40/12.5 mg/d (weeks 7-9) and OM/HCTZ 40/25 mg/d (weeks 10-12) were started if blood pressure (BP) remained > or =120/80 mm Hg at each time interval. End points were change from baseline in mean systolic BP (primary) and mean diastolic BP (secondary). OM-based treatment was well tolerated and changed BP by -22.3/-12.1 mm Hg from baseline vs -0.1/+0.8 mm Hg for placebo (P<.0001). Cumulative goal BP (<140/90 mm Hg) was achieved in 74.1% and 30.7% of OM- compared with placebo-treated patients, respectively (P<.0001). BP normalized (<120/80 mm Hg) in 44.8% of OM- vs 1.4% of placebo treated patients with stage 1 hypertension (P<.0001). PMID- 19120718 TI - Barriers to antihypertensive medication adherence among adults--United States, 2005. AB - Antihypertensive agents are one of the most commonly prescribed classes of medications in the country, but patient adherence rates are low. To better understand why rates are low, the authors used data from the 2005 HealthStyles survey and found that among the 1432 respondents who received prescriptions for antihypertensive medications, 407 (28.4%) reported having difficulty taking their medication. "Not remembering'' was the most common reason reported (32.4%), followed by cost (22.6%), having no insurance (22.4%), side effects (12.5%), other reasons (12.3%), not thinking there is any need (9.3%), and having no health care provider (4.7%). In a multivariate model, younger age, lower income, having mental function impairment, and having had a blood pressure check more than 6 months earlier were factors significantly associated with reporting difficulty taking prescribed antihypertensive drugs. Control of hypertension is a significant public health issue, and alleviating barriers to medication adherence should be a major goal toward hypertension management. PMID- 19120719 TI - Early antihypertensive efficacy of olmesartan medoxomil. AB - The authors assessed the early antihypertensive efficacy of olmesartan medoxomil (OM) in a 12-week prospective observational study. Of 2221 patients with untreated hypertension who received OM (mainly 10 or 20 mg), 331 patients whose blood pressure (BP) was measured at 1 week after initiation of treatment were defined as the "early BP determination group,'' whereas the remaining 1890 patients were defined as the ;;standard BP determination group.'' Baseline characteristics, doses of OM, concomitant drugs used, and BP during treatment did not differ between the 2 groups. The achievement rate of BP target (<140/90 mm Hg) was 28.4% at 1 week in the early BP determination group and 28.3% at 2 weeks in the standard BP determination group (P=NS). Rates of adverse drug reactions in the 2 groups were similar. The present study suggests that OM is safe and effective in reducing BP at an early time point of treatment. PMID- 19120720 TI - Cyclic rapid fluctuation of hypertension and hypotension in pheochromocytoma. PMID- 19120721 TI - Intracerebral hemorrhage as the presenting feature of concurrent pheochromocytoma and paragangliomas. PMID- 19120722 TI - Should the heart rate of hypertensive patients influence clinical decisions? PMID- 19120723 TI - Can an angiotensin receptor blocker be used in a patient in whom angioedema developed with an angiotensin-converting enzyme inhibitor? PMID- 19120724 TI - Treatment of systolic hypertension and severe asymptomatic aortic regurgitation. PMID- 19120725 TI - Analysis of recent papers in hypertension. PMID- 19120726 TI - Embracing the theory/practice gap. PMID- 19120727 TI - A review of quality assessment of the methodology used in guidelines and systematic reviews on oral mucositis. AB - AIMS AND OBJECTIVES: The objective of this study was to identify and to assess the quality of evidence-based guidelines and systematic reviews we used in the case of oral mucositis, to apply general quality criteria for the prevention and treatment of oral mucositis in patients receiving chemotherapy, radiotherapy or both. DESIGN: Systematic review. METHODS: Literature searches were carried out in several electronic databases and websites. Publications were included if they concerned oral mucositis involving adults treated for cancer and had been published after 1 January 2000. As far as systematic reviews were concerned, the article had to report a search strategy, if the search was minimally conducted in the database PubMed or Medline and the articles included in the review were subjected to some kind of methodological assessment. The Appraisal of Guidelines for Research and Education (AGREE) instrument was used to assess the quality of the guidelines and the Overview Quality Assessment Questionnaire (OQAQ) was used for the quality of systematic reviews. RESULTS: Thirty-one articles met the inclusion criteria of which 11 were guidelines and 20 were systematic reviews. Nine of the 11 guidelines did not explicitly describe how they identified, selected and summarised the available evidence. Reviews suffered from lack of clarity, for instance, in performing a thorough literature search. The quality varied among the different guidelines and reviews. CONCLUSION: Most guidelines and systematic reviews had serious methodological flaws. RELEVANCE TO CLINICAL PRACTICE: There is a need to improve the methodological quality of guidelines and systematic reviews for the prevention and treatment of oral mucositis if they are to be used in clinical practice. PMID- 19120728 TI - Systematic review on the effectiveness of caffeine abstinence on the quality of sleep. AB - AIM: The aim of the present study is to review the effects of caffeine abstinence on the quality of sleep. BACKGROUND: Insomnia is a common problem and abstinence from caffeine is the most popular component in sleep hygiene advice. However, there have been inconsistent results relating to the effectiveness of caffeine abstinence in improving sleep. DESIGN: Systematic review. METHODS: We browsed several electronic databases and reference lists of articles about the correlation of caffeine consumption and sleep deprivation. We selected the articles according to predefined inclusion and exclusion criteria. Two reviewers assessed the quality of trials, which were selected according to the Jadad quality assessment scale. We included the trials scoring three or above in the systematic review and extracted their data. We assessed the heterogeneity of the studies before we considered whether or not to combine the studies' results. RESULTS: Three randomised control trials fulfilled the selection criteria among which two trials scored > or =3 on the Jadad scale. We included these two trials in our systematic review. The designs and outcome measurements of these two trials were not homogeneous, therefore, we did not combine their results. Instead, we conducted a critical appraisal. In one trial, caffeine abstinence was associated with significant lengthening of sleep duration (p < 0.01) and better sleep quality (p < 0.05). In another trial, subjects had less difficulty falling asleep on days when they drank decaffeinated coffee (p < 0.05). CONCLUSIONS: The results showed that caffeine abstinence for a whole day could improve sleep quality. Thus, health practitioners were recommended to include caffeine abstinence in the instructions for sleep hygiene. RELEVANCE TO CLINICAL PRACTICE: This study demonstrates the effectiveness of caffeine abstinence in improving sleep quality. It provides evidence for the practice of including caffeine abstinence in sleep hygiene advice. PMID- 19120730 TI - Factors related to incidence of eye disorders in Korean patients at intensive care units. AB - AIMS: The purpose of this study was to identify the incidence rate of eye disorders and further to identify factors related to incidence of eye disorders in intensive care unit patients. BACKGROUND: Although the incidence of eye disorders in intensive care unit patients is very high, it is difficult to come to an accurate estimate of number of eye disorders in intensive care unit patients because eye disorders are often perceived to be minor problems. DESIGN: A retrospective, descriptive survey design was used. METHOD: The medical records of a total 235 patients with eye disorders were identified through a review of the medical records of 2,500 patients hospitalised in the intensive care units of Yonsei University Hospital, Seoul, Korea from January to December, 2004. To examine factors related to eye disorders, 522 patients without eye disorders from the cohort of 2,265 patients were randomly selected. Factors related to incidence of eye disorder were included in a multiple logistic regression model, after screening by the chi-squared test. RESULTS: The incidence rate for eye disorders in the intensive care unit patients was 8.6%. From the multiple regression model, the following odds ratios (95% confidence intervals) of eye disorder were estimated; length of stay in intensive care unit of at least seven days: 2.8 (1.70-4.70); death: 2.5 (1.47-4.29); drowsy mental state: 2.2 (1.10-4.37); stupor mental state: 7.0 (3.20-15.45); coma mental state: 10.8 (3.47-33.74); no self respiration: 1.9 (1.00-3.52); positive end expiratory pressure: 2.9 (1.66-4.92); sedatives: 4.2 (2.26-7.74); muscle relaxants: 2.3 (1.11-4.95). CONCLUSIONS: Factors related to incidence of eye disorders in intensive care unit patients identified in this study support the need to pay attention to eye problems and eye care in intensive care unit patients. RELEVANCE TO CLINICAL PRACTICE: An evidence-based eye care protocol should be provided as routine care to prevent eye complication, especially in critically ill patients with mechanical ventilators, positive end expiratory pressure, sedatives or muscle relaxants and for patients whose mental status is decreased. PMID- 19120729 TI - Removal of oral secretion prior to position change can reduce the incidence of ventilator-associated pneumonia for adult ICU patients: a clinical controlled trial study. AB - AIM: The purpose of this study was to explore the effect of oral secretion on aspiration and reducing ventilator-associated pneumonia. BACKGROUND: Ventilator associated pneumonia is a serious hospital-acquired infection with reported incidence rate of 12.2% and mortality rate of 29.3%. Oral secretion is purported as a media which brings the oropharyngeal pathogens down to the respiratory track. METHODS: Two-group comparison study design was adopted. Subjects were recruited from an adult general intensive care unit of a medical centre in Taipei city. Patients in the study group received suction of oral secretion before each positional care, in contrast with patients in the control group who received routine care. RESULTS: Ventilator-associated pneumonia was found in 24 of 159 (15.1%) patients in the control group and in five of 102 (4.9%) patients in the study group with a reduction of risk ratio of 0.32 (95% CI 0.11-0.92). Eight of the 24 ventilator-associated pneumonia patients died in the control group; however, none of those ventilator-associated pneumonia patients died in the study group. The increased chance of survival was 1.50 (95% CI 1.13-1.99). The length of stay in ICU and duration of mechanical ventilation were reduced in the study group. In consideration of cost, the cost of tubes used to remove oral secretion is much less than the one used to do continuous subglottal suction. CONCLUSION: Removal of oral secretion is effective in reducing the incidence of ventilator associated pneumonia with minimum cost intervention. RELEVANCE TO CLINICAL PRACTICE: This study provides evidence that removal of oral secretion prior to position change is cost effective to reduce the incidence of ventilator associated pneumonia. As such intervention is an easy task, routine removal of oral secretion is recommended as the standard of daily nursing care of patients on ventilator. PMID- 19120731 TI - Misconceived and maladaptive beliefs about heart disease: a comparison between Taiwan and Britain. AB - AIMS AND OBJECTIVES: To assess and compare misconceived and maladaptive beliefs about coronary heart disease between Taiwanese and British people with heart disease. BACKGROUND: Holding misconceived and maladaptive beliefs about heart disease has deleterious effects on a patient's quality of life. Cultural contexts influence a person's responses to illness, but little information exists about the ways in which cultural values influence a person's attributions and coping behaviours regarding their heart disease. DESIGN: A cross-sectional survey using a descriptive comparative design was carried out in Taiwan and Britain. METHODS: A structured questionnaire was used to elicit misconceived or potentially maladaptive beliefs about heart disease. People with a confirmed diagnosis of coronary heart disease were recruited from a teaching hospital in Taipei (n = 238) and a tertiary cardiothoracic centre in the North of England (n = 204). RESULTS: Taiwanese patients held more misconceived and maladaptive beliefs about heart disease than those in Britain. Both British and Taiwanese patients showed high agreement on 'stress is one of the main causes of heart disease' and 'always avoid stress', but these beliefs were more common in Taiwanese patients (p < 0.001). Misconceptions about avoiding anything that might bring on angina were common in both countries. CONCLUSIONS: While there were some similarities in the cardiac beliefs held by the two groups of patients, Taiwanese patients held significantly more misconceptions. This may be explained by differences between the patients' cultures, including social norms, health care provision and health information from the media. RELEVANCE TO CLINICAL PRACTICE: Interventions should be developed to dispel maladaptive beliefs thereby altering the coping actions of patients. The meaning and purpose of specific behaviours needs to be interpreted within the context of a patient's culture. Awareness of cultural perspectives may help to develop nursing care plans that are more successful as they are more individually applicable to patients. PMID- 19120732 TI - 'A pacemaker in my heart' -- classification of questions asked by pacemaker patients as a basis for intervention. AB - AIMS: The research aimed to identify the information patients find necessary, following pacemaker implantation. BACKGROUND: Although pacemaker devices do not have an adverse impact on lifestyle, they evoke anxiety related to the patient's activities and lifestyle. DESIGN: Survey. METHODS: A convenience sample of participants was taken from the entire population of patients who attended the cardiology clinic between January-June 2007; 274 individual meetings were held with 123 pacemaker patients in three periods, reflecting different stages of recovery. In the meeting, patients were invoked to ask any question they may have regarding pacemaker implantation. The questions were collected from the patients and categorised chronologically, in accordance with their frequency in three periods, reflecting different stages of recovery. RESULTS: Eight categories, representing common issues and content were raised: motion and effort, environmental influences, personal hygiene, knowledge concerning the pacemaker operation, medical treatment, eating and drinking, clothing and general questions. Findings show that the common factor for most of the questions was the loss of confidence in the various aspects of life. The largest relative question proportion was in the motion and effort (27%) (e.g. may I swim? how many kg may I lift?) and environmental influences (26%) (e.g. may I use a cellular phone? may I use a shaving machine?). CONCLUSIONS: A coherent, continuous pattern was found, characterising the different points of measurement where, at the first point, questions were characterised as more existential, related to daily routine activities and as time passed and patients were exposed to non-daily activities and conditions, other questions were raised. RELEVANCE TO CLINICAL PRACTICE: Pacemaker implant guidance should be based upon experimental evidence, arranged according to each category's relative weight and take into account the patient's point of view. PMID- 19120733 TI - Coronary heart disease patients' perception of their symptoms and sense of control are associated with their quality of life three years following hospital discharge. AB - BACKGROUND: Recent intervention studies have attempted to address cardiac rehabilitation patients' illness beliefs to improve care. Their findings were mainly limited to short-term behaviour changes such as return to work or attendance on rehabilitation programmes. Evidence about what aspects of the patients' illness beliefs influence long-term quality of life in cardiac rehabilitation remains sparse. OBJECTIVE: To examine the association between coronary heart disease patients' illness beliefs and their quality of life three years after hospital discharge. DESIGN: A longitudinal prospective survey. METHODS: Coronary heart disease patients (n = 253) completed validated questionnaires at three points. Patients' illness beliefs were measured twice: before hospital discharge and nine months after discharge - using the Illness Perception Questionnaire. Quality of life was assessed as the outcome measure at three years after discharge, using the Health Survey SF36v2 questionnaire. Demographic, illness characteristics and cardiac rehabilitation attendance were also assessed. RESULTS: Two multiple regression analyses were performed with each of the SF36v2 physical and mental health summary scores as outcome variables. Analysis of baseline data showed 43% of the variance in the physical health summary score at three years was accounted for by a perceived lower number of symptoms, belief that the illness was controllable, admission as an emergency and no previous history of cardiac illness; 22% of the variance in the mental health summary score was accounted for by perceived lower number of symptoms and no previous history of cardiac illness. Repeat analysis using the data collected nine months after discharge showed similar findings. CONCLUSION: Coronary heart disease patients' perception of their symptoms and sense of control at time of discharge were significantly associated with their quality of life three years after discharge. RELEVANCE TO CLINICAL PRACTICE: This study promotes the use of the assessment of patients' illness beliefs as a routine practice to improve care. PMID- 19120734 TI - Sickness-related dysfunction in persons with self-reported multiple chemical sensitivity at four levels of severity. AB - AIM: To examine quality of life outcome for persons who self-report chemical sensitivity, often referred to as multiple chemical sensitivity. BACKGROUND: Multiple chemical sensitivity is poorly understood with few providers specialising in its treatment. This lack of treatment and the ubiquity of chemicals engender severe life impacts such as job loss, financial loss, social isolation and even homelessness for persons who experience these sensitivities. DESIGN: Survey. METHOD: We examined chemical incitants, symptoms and sickness related behavioural dysfunction as measured by the Sickness Impact Profile in 254 persons self-identified with multiple chemical sensitivity. RESULTS: Chemicals rated as causing the most symptomatology in respondents were pesticide, formaldehyde, fresh paint, new carpet, diesel exhaust, perfume and air fresheners. The five highest rated symptoms in this sample were tiredness/lethargy, difficulty concentrating, muscle aches, memory difficulties and long-term fatigue. Overall mean Sickness Impact Profile score was 25.25%, showing serious impairment, with the most serious dysfunction in the categories of work (55.36%), alertness behaviour (53.45%) and recreation and pastimes (45.20%). CONCLUSION: Multiple chemical sensitivity is an important health care issue because it often includes serious dysfunction, is poorly understood by providers and poses extensive financial and treatment obstacles for those who experience it. RELEVANCE TO CLINICAL PRACTICE: Persons with multiple chemical sensitivity seek medical treatment in a variety of contexts and informed providers can both avoid iatrogenic harm due to medical exposures and provide any possible treatment for the chemical sensitivities. Understanding the impact of the health condition is crucial to communicate with and treat persons who experience the sensitivities. PMID- 19120735 TI - Predicting pressure ulcer risk: a multifactorial approach to assess risk factors in a large university hospital population. AB - AIMS: The purpose of this study was: (1) to determine the combination of risk factors which best predicts the risk of developing pressure ulcers among inpatients in an acute care university hospital; (2) to determine the appropriate weight for each risk factor; and (3) to derive a concise and easy-to-use risk assessment tool for daily use by nursing staff. BACKGROUND: Efficient application of preventive measures against pressure ulcers requires the identification of patients at risk. Adequate risk assessment tools are still needed because the predictive value of existing tools is sometimes unsatisfactory. DESIGN: Survey. METHODS: A sample of 34,238 cases admitted to Essen University Clinics from April 2003 and discharged up to and including March 2004, was enrolled into the study. Nursing staff recorded data on pressure ulcer status and potential risk factors on admission. Predictors were identified and weighted by multivariate logistic regression. We derived a risk assessment scale from the final logistic regression model by assigning point values to each predictor according to its individual weight. RESULTS: The period prevalence rate of pressure ulcers was 1.8% (625 cases). The analysis identified 12 predictors for developing pressure ulcers. With the optimum cut-off point sensitivity and specificity were 83.4 and 83.1%, respectively, with a positive predictive value of 8.4% and a negative predictive value of 99.6%. The diagnostic probabilities of the derived scale were similar to those of the original regression model. CONCLUSIONS: The predictors mostly correspond to those used in established scales, although the use of weighted factors is a partly novel approach. Both the final regression model and the derived scale show good prognostic validity. RELEVANCE TO CLINICAL PRACTICE: The derived risk assessment scale is an easy-to-understand, easy-to-use tool with good prognostic validity and can assist in effective application of preventive measures against pressure ulcer. PMID- 19120736 TI - Injured road users' health-related quality of life after telephone intervention: a randomised controlled trial. AB - OBJECTIVE: To investigate whether nursing intervention via telephone follow-up can affect health-related quality of life among road users in the postinjury phase. BACKGROUND: Injuries in the traffic environment often cause both physical impairment and psychological trauma. Therefore, it is important to investigate whether nursing via telephone follow-up after discharge could affect health related quality of life among injured road users. DESIGN: A randomised controlled study. METHOD: Between April 2003-April 2005, car occupants, cyclists and pedestrians, aged 18-70 years, all injured in the traffic environment, were randomly assigned to an intervention (n = 288) or control group (n = 280). All patients were initially managed by one emergency department with the intervention group being followed up by nurse led telephone follow-up three weeks after discharge, while the control group did not receive any telephone follow-up. Data were collected from a mail survey using the non-disease-specific health-related quality of life instrument EQ5D, at baseline and after six months. RESULTS: In general, the intervention group rated significantly higher health-related quality of life than the control group. This difference was most pronounced in the group of those provided with advice as part of telephone follow-up and significantly for the general health status. Car occupants gained most advantage from the nursing telephone follow-up, with significantly lower problems in the dimensions of pain/discomfort and usual activities. CONCLUSION: Nursing intervention via telephone follow-up was effective at increasing the health-related quality of life of injured road users. Early identification of the concerns of the injured and specific advice provided by the intervening nurse seems to be of greatest weight. Thus, more research is needed regarding the usefulness of early support and the advantage of this from a health economic perspective. RELEVANCE TO CLINICAL PRACTICE: There is a need for changes in attitudes and working routines to identify individuals with low health-related quality of life and to give more extensive support from this point of view. PMID- 19120737 TI - Single ability among activities of daily living as a predictor of agitation. AB - AIMS: The aims of this study were to investigate whether the interaction effect between restraint and functional ability, and single ability during activities of daily living can predict agitation among residents with dementia. BACKGROUND: Agitated behaviour is one of the most common symptoms of dementia and might endanger the patients themselves, caregivers and institutions. However, the prevalence of problem behaviours and its associated factors at long-term care facilities in Taiwan are less understood. DESIGN: A direct observation was used to observe the agitated behaviours of residents with dementia in special care units. METHODS: Residents of dementia special care units who were diagnosed with dementia in eight long-term care facilities were recruited. Measurements included: demographic data, the Barthel Index, the Mini-Mental State Examination and the Cohen-Mansfield Agitation Inventory. RESULTS: The number of subjects who were identified with problem behaviours was 163 (43.5%). Significant differences in sex, being restrained, restrained time, age, family visits, functional status and mental status were found between the agitated and non-agitated groups. However, mental status, family visits, walking ability, being restrained and getting in and off toilet were five independent factors associated with agitated behaviours after controlling for all other factors. CONCLUSIONS: It is recommended that strategies be constructed to encourage the family to periodically visit older residents and to develop restraint-free environments in long-term care facilities. RELEVANCE TO CLINICAL PRACTICE: Residents with dementia require significant daily living support and behaviour management as their illness progress. PMID- 19120738 TI - Understanding inadequate pain management in the clinical setting: the value of the sequential explanatory mixed method study. AB - AIM: The purpose of this paper is to critically explore the sequential explanatory mixed method research design and how it can enhance our understanding of pain management. BACKGROUND: The general prevalence of pain after surgery has not changed significantly over several decades despite the widespread introduction of new pain relieving technologies. The majority of postoperative pain studies use quantitative methods which offer little understanding of the underlying processes of care. Understanding can be illuminated by using an explanatory mixed method research design. DESIGN: Discursive paper. METHOD: This paper focuses on the methodological considerations when using a mixed method design. Two previously published mixed methods studies illustrate how findings can inform practice. In the first, 85 women undergoing surgery completed questionnaires to measure pain, anxiety and depression. Telephone interviews explored their pain experiences. The second study considered frequency and patterns of anxiety in the immediate pre and postoperative period. Semi structured telephone interviews, identified contributing events/situations amenable to nursing intervention. DISCUSSION: Reasons for growing popularity, criticisms, paradigmatic considerations and epistemological roots of pragmatism are explored. The two explanatory mixed method studies provide examples of these studies and how 'inferences' from quantitative and qualitative data can inform practice. CONCLUSION: This paper connects quantitative and qualitative data, drawing on two research studies, to give greater understanding to the management of pain. Knowledge of the processes responsible for inadequate pain management can be illuminated by using explanatory mixed methods research designs. RELEVANCE TO CLINICAL PRACTICE: Nursing requires knowledge which reflects the complexity of human health. The explanatory mixed method study can elucidate the problem under scrutiny, e.g. prevalence of pain or anxiety. The qualitative phase can generates an understanding of contributing factors and insights for care delivery. The implicit desire to change and influence practice makes it relevant for those closely aligned to practice. PMID- 19120739 TI - The use of physical restraint in institutional care of older people in Finland: nurses' individual, communal and alternative modes of action. AB - AIMS AND OBJECTIVES: The aim of the study was to map nursing staff's individual, communal and alternative modes of action in situations where they used physical restraint of older people in Finland. BACKGROUND: The use of physical restraint in institutional care of older people involves modes of action that are linked to the personalities and modes of operation of individual nurses or to communal modes of operation mutually agreed on in the workplace. Nurse's individual modes of operation are linked to consideration towards older people when using physical restraint. Communal modes of operation include joint discussion among nurses, decision-making and written guidelines concerning the use of physical restraint. DESIGN: A quantitative survey. METHODS: The data were collected from nurses (n = 1148) working in healthcare centre wards, municipal and private nursing homes in Finland using a structured questionnaire (response rate 78%). RESULTS: Older nurses and those with the longest working experience were most active in using individual modes of operation in situations where physical restraint was applied. Nurses' participation in continuing education in care of older people increased the use of both individual and alternative modes of action. The alternative modes of action were one that understands the older person and one that focuses on negotiation. The use of physical restraint was often discussed in the workplace during information sessions or during work, but only 33% had written guidelines on the use of physical restraint. CONCLUSIONS: Education can be used to increase nurses' ability to use alternative modes of action and thus decrease the use of physical restraint in institutional care of older people. Nursing management has a key role in making sure that units caring for older people receive written guidelines on the use of physical restraint. RELEVANCE TO CLINICAL PRACTICE: Locums especially need more education to avoid the use of physical restraint. PMID- 19120740 TI - A developmental approach to the treatment of bipolar disorder: IPSRT with an adolescent. AB - AIM: This case study explains how a psychotherapy previously used with adults can be used with adolescents by focusing on the specific developmental issues associated with adolescence. BACKGROUND: Bipolar disorder is a damaging disorder to experience during the developmental phase of adolescence. Interpersonal social rhythm psychotherapy has been developed as an adjunct to medication for managing bipolar disorder and shows some promising outcomes in adults. DESIGN: This is a single case study design drawn from a larger randomised control trial of two psychotherapies for bipolar disorder. The case study addressed the question: How can Interpersonal social rhythm therapy be applied with adolescents who have bipolar disorder? METHOD: This study used a purposeful sampling process by selecting the youngest adolescent participating in the randomised control trial. All the subject's sessions of Interpersonal social rhythm therapy were taped, transcribed and analysed. The analysis involved describing the process of psychotherapy as it occurred over time, mapping the process as a trajectory across the three phases of psychotherapy experience and focusing the analysis around the impact of bipolar disorder and IPSRT on adolescent developmental issues, specifically the issue of identity development. RESULTS: Interpersonal social rhythm therapy allowed the therapist to address developmental issues within its framework. As a result of participation in the psychotherapy the adolescent was able to manage her mood symptoms and develop a sense of identity that was age-appropriate. CONCLUSION: Interpersonal social rhythm therapy provided the adolescent in the case study the opportunity to consider what it meant to have bipolar disorder and to integrate this meaning into her sense of self. RELEVANCE TO CLINICAL PRACTICE: Bipolar disorder is a chronic and recurring disorder that can have a serious impact on development and functioning. Interpersonal social rhythm therapy provides an approach to nursing care that enables adolescents to improve social functioning. PMID- 19120741 TI - Commentary on Birch D and Draper J (2008) A critical literature review exploring the challenges of delivering effective palliative care to older people with dementia. Journal of Clinical Nursing 17, 1144-1163. PMID- 19120742 TI - Commentary on Allan H (2007) The rhetoric of caring and the recruitment of overseas nurses: the social production of a care gap. Journal of Clinical Nursing 16, 2204-2212. PMID- 19120743 TI - Commentary on Ayaz S & Kubilay G (2009) Effectiveness of the PLISSIT model for solving the sexual problems of patients with stomas. Journal of Clinical Nursing 18, 89-98. PMID- 19120744 TI - Editorial: Practice development: who cares and so what? PMID- 19120745 TI - Editorial: Practice development: 'to be what we want to be'. PMID- 19120746 TI - Metasynthesis: withdrawing life-sustaining treatments: the experience of family decision-makers. AB - AIM: The present study was undertaken to synthesise findings from qualitative investigations of family participation in decisions to withdraw and/or withhold life-sustaining treatment from a seriously ill family member. BACKGROUND: As a consequence of increasing effectiveness and sophistication of available medical interventions, death is commonly preceded by a decision to withdraw or withhold potentially life-sustaining treatments. These decisions take place in a bioethical context characterised by the pre-eminence of self-determination, but patients are typically too ill to make their own decisions. Thus, family members are called upon to participate in these morally consequential decisions on the patient's behalf. DESIGN: Metasynthesis. METHOD: Metasynthesis is a form of inquiry that provides for integration of qualitative studies' findings to strengthen knowledge for practice and advance theoretical development. This metasynthesis was conducted using the constant comparative methods of grounded theory. RESULTS: Family members engage in a process of participation in decision making that is comprised of three major categories: reframing reality, relating and integrating. Surrogates used both cues and information as they reframed their understanding of the patient's status. Relationships with providers and with other family members powerfully influenced the decision-making process. Integrating was characterised by reconciling and going forward. This part of the process has both intrapersonal and interpersonal aspects and describes how family decision-makers find meaning in their experience and move forward in their lives. CONCLUSIONS: This synthesis provides a more comprehensive and empirically supported understanding of family members' experiences as they participate in treatment decisions for dying family members. Relevance to clinical practice. This metasynthesis provides evidence to improve family care during treatment withdrawal/withholding decision-making and a theoretical model that can be used to guide creation of clinical practice guidelines. Through increased understanding of family members' experiences, clinicians can more effectively support family decision-making processes. PMID- 19120747 TI - Living with a relative who has a spinal cord injury: a grounded theory approach. AB - AIMS: This paper reports a study which explored family carers' experiences of taking care of a relative who has a spinal cord injury. BACKGROUND: Spinal cord injury is one of the most disastrous injuries a person may experience. Although one family member experiences the injury, the entire family is affected. In Taiwan, family carers carry the primary responsibility of providing care on a 24 hour basis during the rehabilitation hospitalisation. However, very limited research exists regarding their experiences. DESIGN: This qualitative study has a cross-sectional and descriptive-explorative design. METHOD: Grounded theory was used to explore the psychosocial implications for family carer who has a relative with spinal cord injury. Data collection and analysis. Data were collected through in-depth, tape-recorded, semi-structured interviews and observation of a group discussion and two religious activities in a rehabilitation hospital in Taiwan. Fifteen family carers participated in this study comprising eight individual interviews and seven participating in the group discussion. Data collection and analysis occurred concurrently. RESULTS: The findings resulted in the core category 'living with a relative who has a spinal cord injury' and identified the experience of taking care of a relative who has a spinal cord injury. Three stages, including four categories: stage 1 'A catastrophic life event'; stage 2 'Confronting challenges'; and stage 3 'Family resilience' or 'Family breakdown'. CONCLUSIONS: This study shows that family carers experience a catastrophic life event, they have to confront challenges and the positive consequence is family resilience, the negative consequence is family breakdown. Relevance to clinical practice. Knowing how prospective family carers view their own situation and what they need is essential to provide effective nursing care for the family that has been changed dramatically by a spinal cord injury. PMID- 19120748 TI - Qualitative analysis of patients' intensive care experience during mechanical ventilation. AB - AIM: To understand patients' intensive care experience while receiving mechanical ventilation in intensive care units. BACKGROUND: The mechanically ventilated patient's experience in the intensive care unit is unique. Notably lacking are international studies on patients' experience, particularly those living in Asia. A better understanding of patients' experience is needed for nurses to develop approaches to take care of these patients. DESIGN: A phenomenological approach formed by the ideas of Heidegger was used. METHODS: Eleven participants surviving from mechanical ventilation were interviewed in-depth. Patients were asked to describe their experience by responding to the question 'what is it like to experience mechanical ventilation treatment at an ICU?'. Giorgi's phenomenological analysis procedure was used to analyse the data. RESULTS: Five mutually exclusive themes emerged, which were: 'being in an unconventional environment', 'physical suffering', 'psychological suffering', 'self encouragement' and 'self-reflection'. 'Self-encouragement' and 'self-reflection' enhanced patients' self-confidence, which was beneficial to recovering. CONCLUSION: Patient's experiences while receiving mechanical ventilation in the intensive unit were poignant and frightening. RELEVANCE TO CLINICAL PRACTICE: Critical care nurses should place the highest priority on recognising and meeting the needs of ventilated patients in intensive care units. An intensive care unit should be a place for the patient to live as a human being; not just a place to survive. PMID- 19120750 TI - Leadership as part of the nurse consultant role: banging the drum for patient care. AB - AIMS AND OBJECTIVES: This paper draws upon an evaluation of the first group of nurse consultants in Scotland. The evaluation aimed to identify the extent to which they fulfilled the remit of their posts which comprised four core functions. One of these functions was to provide professional leadership and this paper focuses on this element of the role and aims to explore it in relation to the attributes of transformational leadership. BACKGROUND: Nurse consultants were introduced in the UK in 2000. Their purpose was to achieve better outcomes for patients and strengthen leadership. Nursing research identifies leadership as a key element of the role, with postholders adopting transformational leadership approaches. Research from the fields of sociology and psychology identifies difficulties in formulating a coherent theory of leadership, arguing for better understanding of leadership processes. DESIGN: Qualitative. METHODS: This paper draws on 31 semi-structured interviews with four nurse consultants who were interviewed twice over six to nine months and 23 other 'stakeholders' who worked with them. RESULTS: Varied leadership activity at ward, NHS Trust and strategic levels was identified. Postholders used approaches that resonated with the attributes of transformational leadership. Leadership processes included developing a vision for the service, acting as mediator and champion, and exerting control over complex change initiatives. Techniques of leadership included taking a 'softly softly' approach, pacing change initiatives and arguing assertively with those in senior positions. Interview findings also identified the level of preparation that was required to meet the remit of the posts, highlighting the importance of interpersonal skills and intellectual effort in achieving outcomes. CONCLUSIONS: Nurse consultants require considerable technical expertise, cognitive and interpersonal skills, and the ability to take risks. The data suggest that the leadership attributes required are transformational in nature but that they also exceed those identified in much of the literature. These posts require appropriate support if they are to be sustainable. Relevance to clinical practice. This paper adds to our understanding of the complexity of the nurse consultant role and highlights the challenge of providing appropriate professional development for postholders. PMID- 19120749 TI - Willingness to use respite care among family caregivers in Northern Taiwan. AB - AIM AND OBJECTIVES: Investigate the use of respite care among family caregivers and examine factors related to their willingness to use such care. BACKGROUND: Respite care is a pivotal service for supporting family caregivers. Although research related to respite care is not a new field in western societies, such studies have rarely been done for Asian populations. DESIGN: A cross-sectional study. METHODS: The study was conducted using a convenience sample of 100 family caregivers obtained through four home health care agencies in Northern Taiwan. Structured face-to face interviews were conducted in home settings. Caregivers were divided into a willingness group and a non-willingness group. The quantitative data were analysed using t-test, chi-square and multiple logistic regression. RESULTS: Sixty-five per cent of the caregivers interviewed did not know about respite care services. The prevalence of respite care use was 11%. Eighty-nine per cent of the caregivers had never used respite care: 60.7% of them reported that they would never use it and 39.3% of the caregivers indicated that they were willing to use respite care in the future. This study showed that willingness to use respite care was higher when the respite services matched the needs of the caregivers as well as the caregivers perceived higher caregiving burden. CONCLUSIONS: These results suggest that respite care services should be explained more explicitly to family caregivers. Also, any respite care plan must consider the family caregivers burden, both perceived and realistically. RELEVANCE TO CLINICAL PRACTICE: The willingness to use respite care is a culturally sensitive issue. Health care providers must act as advocates to improve respite care policy. They must ensure that respite care fits caregivers' needs and they must help caregivers overcome the social or cultural barriers to respite care. PMID- 19120751 TI - Nurse prescribers' interactions with and perceptions of pharmaceutical sales representatives. AB - AIMS AND OBJECTIVES: The aim of our study was to investigate the perceptions of pharmaceutical sales representatives by nurse prescribers. BACKGROUND: Nurses with advanced training have earned prescriptive authority in North America, Europe and other parts of the world. These nurses are being increasingly targeted by pharmaceutical sales representatives. There is a paucity of data regarding nurses' perceptions of pharmaceutical sales representatives. DESIGN: Survey. METHODS: A convenience sample of nurse prescribers was recruited to complete an Internet questionnaire about their interactions with and perceptions of sales representatives. The data were collected over one month ending in January 2007. There were 39 survey items ranging from perception-based items assessed by Likert type scale to open-ended queries. Descriptive statistics were used to summarise the results. RESULTS: Ninety-two nurses completed this survey, which demonstrated good internal consistency yielding a Cronbach's alpha coefficient of 0.83. Positive perceptions of pharmaceutical representatives included: explaining their products clearly (80.4%) and knowledge about their medications (88.0%). Negative aspects included: lack of consideration of nurses' time (50%) and failure to equally discuss medication strengths and weaknesses (21.8%). Perhaps the most alarming finding was that 35.9% of respondents indicated that sales representatives suggested paybacks for promoting their drugs. CONCLUSIONS: Nurses with prescriptive authority generally perceive interactions with pharmaceutical sales representatives as positive. However, they also have concerns about the nature and methods of some of their activities. Nations that have nurses with prescribing authority can benefit from observing both the mis-steps and the positive inroads that have already been made by the profession in the USA and other countries. RELEVANCE TO CLINICAL PRACTICE: Appropriate use of pharmaceutical sales representatives' services may enhance the ability of nurse prescribers to deliver optimal nursing care. Methods, such as counter-detailing may be necessary to maintain an evidence-based approach as the controlling factor. PMID- 19120752 TI - Understanding the practical and theoretical development of social rehabilitation through action research. AB - AIM: This paper aims to present and reflect on the process of practical and theoretical change in an action research project in which the issue of social rehabilitation was developed. BACKGROUND: Action research is a useful method to change and advance practice. Consequently, grounded knowledge, which forms the basis of the practical change, is developed. 'Social rehabilitation' was the field of clinical practice which needed enhancement as the literature lacked nurse-led social rehabilitation programmes in the neurology field. DESIGN: This was a cyclic action research project. METHODS: The project took place in a highly specialised hospital in Spain and aimed to promote social life of neurological patients and relatives through the planning, implementation and evaluation of a nurse-led social rehabilitation programme based on individual needs. In this project, which lasted 30 months, multiple triangulation of sources and methods was applied. Thirty-seven nurses participated and 22 and 18 patients and their relatives constituted a baseline and an intervention group, respectively. RESULTS: Several issues were carefully treated in this action research project to develop practical and theoretical knowledge about social rehabilitation: the validity and reliability of the project, the work organisation of the wards, the nurses' attitudes towards new care, the researcher-participant relationship and the controversial role of professionals in social rehabilitation. CONCLUSION: The nurses' emancipation in the process of practice change led to practical and theoretical assimilation of social rehabilitation. Intensive work on practitioners' attitudes and the provision of empirical evidence were key interventions to foster controversial roles and enhance services. RELEVANCE TO CLINICAL PRACTICE: Details about the process of change of nursing practice in response to clients' needs have been provided. Some reflections about how to integrate social care and knowledge about social rehabilitation in clinical daily practice are made. PMID- 19120753 TI - A longitudinal study of stress and psychological distress in nurses and nursing students. AB - AIM: The aim of this study was to investigate how differences in life events and stress contribute to psychological distress in nurses and nursing students. BACKGROUND: Stress is an issue for nursing students and qualified nurses leading to psychological distress and attrition. DESIGN: A longitudinal study using four time waves was conducted between 1994-1997. METHODS: Measures were taken of stress, life events and psychological distress in addition to a range of demographic data. Data were analysed using descriptive statistics, linear modelling and mixed-effects modelling. The study was set in Scotland, UK and used newly qualified nurses and nursing students from four university departments of nursing over four years. The study was initiated with 359 participants (147 nurses and 212 nursing students) and complete data were obtained for 192 participants. RESULTS: Stress levels, psychological distress and life events are all associated within time and across time. At baseline, life events and stress contributed significantly to psychological distress. The pattern of psychological distress differed between the nursing students and the newly qualified nurses with a high level in the nurses after qualifying and starting their career. CONCLUSION: Stress, individual traits, adverse life events and psychological distress are all interrelated. Future lines of enquiry should focus on the transition between being a nursing student and becoming a nurse. RELEVANCE TO CLINICAL PRACTICE: Stress and psychological distress may have negative outcomes for the retention of nursing students in programmes of study and newly qualified nurses in the nursing workforce. PMID- 19120754 TI - Sleep quality and morningness-eveningness of shift nurses. AB - AIM AND OBJECTIVE: The aim of the study was to analyse, while controlling for identified covariates, the effects of morningness-eveningness on sleep quality for shift nurses. BACKGROUND: Shift nurses had greater difficulty falling asleep or staying asleep, thus resulting in higher rates of retiring from hospital. Existing research has addressed the effects of manpower demand and personal preferences on shift assignment; however, the concept of endogenous rhythms is rarely considered. METHODS: This analysis included 137 nurses between the ages of 21-58. Nurses completed the Horne and Ostberg questionnaire to assess morningness eveningness and the Pittsburgh Sleep Quality Index (PSQI) questionnaire to measure self-reported sleep quality over the last month. The 18-point Chinese version had a Cronbach's reliability coefficient of 0.79 overall and 0.86 respectively. This study analysed correlates of sleep quality by comparing the groups with better or worse sleep quality according to the median of PSQI. Univariate and multivariate analyses were used for the risk factors of worse sleep quality. RESULTS: The result showed that the strongest predictor of sleep quality was morningness-eveningness not the shift schedule or shift pattern for nurses under controlling the variable of age. Greater age and longer years employed in nursing significantly decreased the risk of worse sleep quality. The confounding age factor was properly controlled; evening types working on changing shifts had higher risk of poor sleep quality compared to morning types. CONCLUSIONS: Morningness-eveningness was the strongest predictor of sleep quality under controlling the variable of age in shift nurses. IMPLICATIONS FOR CLINICAL PRACTICE: Our results suggested that determining if nurses were attributed to morning or evening types is an important sleep issue before deciding the shift assignment. PMID- 19120755 TI - Factors associated with perceived sleep quality of nurses working on rotating shifts. AB - AIMS: To explore nurses' perceived sleep quality and examine factors that contribute to insufficient sleep quality. BACKGROUND: Shift work is an important source of disturbances in the health and well-being of nurses. However, nursing services must be available on a 24-hour basis, making shift work a necessity. Sleep disorders tend to occur among nurses typically working on a rotating schedule. Although many studies related to nurses' sleep quality have been carried out in the West, few have investigated factors linked to nurses' sleep quality in Hong Kong. DESIGN: A cross-sectional study. METHOD: The study was conducted during the period November 2005-June 2006 in two local hospitals in Hong Kong. Nurses (n = 163) completed a self-reported questionnaire. Demographic data and information on health status, strain and symptom levels and perceived sleep quality were collected. RESULTS: More than 70% of the nurses reported having insufficient sleep and strain and symptom levels were higher in this group. Older age, perceived poor sleep status, gastrointestinal symptoms and higher strain and symptom levels were risk factors that contributed to insufficient sleep. CONCLUSIONS: Evaluation of internal stressors and modification of shift work schedules are important areas of future research; these should aim at finding the best compromise between productivity and employees' sleep quality, health and performance. RELEVANCE TO CLINICAL PRACTICE: Healthcare workers' job task analysis, the evaluation of internal stressors and the modification of shift work schedules are important areas of future research and should result in the best compromise between productivity and employees' sleep quality, health and performance. PMID- 19120756 TI - Hepatitis B-Hepatitis C seroprevalences and blunt-penetrating object injuries in housekeepers in Turkey: a survey study. AB - AIM: This study aims to identify housekeepers' use of protective measures, provide data about hepatitis B virus (HBV) and hepatitis C virus (HCV) seroprevalences and HBV immunisation, and investigate blunt-penetrating injuries in patient care services, routine cleaning services and orderly services. BACKGROUND: Hospitals have been described as hazardous work environments with an increase in HBV-HCV seroprevalences and blunt-penetrating object injuries. This situation creates great risks and hazards for housekeepers in their jobs. DESIGN: Survey. METHODS: The study population was housekeepers who work in university hospitals. A total of 824 housekeepers were surveyed by using a 20-item questionnaire. The questionnaire included the sociodemographic characteristics of housekeepers and the risk level of the unit employed. Blood samples were taken from the housekeepers. RESULTS: Their mean age was 32.5 years. The majority of the housekeepers (52.5%) were women and graduates of primary school (51.1%). The mean length of employment was 2.6 years, 73% were working on medical/surgical units, 91.2% were working in routine cleaning and 70.9% had been injured with various blunt and penetrating objects while working in the hospital in the past six months. The obtained result for seroprevalence for HBV-HCV was 2.2%. Only 27.5% of the housekeepers had been immunised with Hepatitis B vaccine. A large percentage of housekeepers in this study had used universal precautions. CONCLUSION: This study showed high seroprevalence rates for HBV-HCV and blunt penetrating object injuries in housekeepers. Therefore, more effort is necessary to increase the use of protective measures against HBV-HCV and blunt-penetrating object injuries in housekeepers. RELEVANCE TO CLINICAL PRACTICE: Hospitals need to take protective measures and implement innovative educational and support programmes organised for specific groups of housekeepers. PMID- 19120757 TI - The Asthma Self-Care Scale: preliminary development and psychometric testing. PMID- 19120758 TI - Commentary on Tannen A, Dassen T and Halfens R (2008) Differences in prevalence of pressure ulcers between the Netherlands and Germany--associations between risk, prevention and occurrence of pressure ulcers in hospitals and nursing homes. Journal of Clinical Nursing 17, 1237-1244. PMID- 19120759 TI - Commentary on Hughes M (2007) Parents' and nurses' attitudes to family-centred care: an Irish perspective. Journal of Clinical Nursing 16, 2341-2348. PMID- 19120760 TI - Commentary on McIntosh and Tolson J D (2009) Leadership as part of the nurse consultant role: banging the drum for patient care. Journal of Clinical Nursing 18, 219-227. PMID- 19120761 TI - Commentary on Banning M (2008) A review of clinical decision making: models and current research. Journal of Clinical Nursing 17, 187-195. PMID- 19120763 TI - Subungual melanoma: histological examination of 50 cases from early stage to bone invasion. AB - Subungual melanoma is a rare form of malignant melanoma. It is extremely difficult to differentiate it histologically from benign melanonychia striata or melanocytic nevus, especially in the early stage. We divided 50 cases of subungual melanoma into four groups according to clinical progress, and examined their histological findings in each respective stage. In the early stage (19 cases), atypical melanocytes were polygonal showing slight nuclear atypia with no mitoses at all. In six out of 19 cases (31.6%), the atypical melanocytes proliferated more in the hyponychium than in the nail matrix, and only very few in the nail bed. Periungual pigmentation (Hutchinson's sign) appeared from the early stage in almost all cases. With stage progression (middle stage, 13 cases; progressive stage, 13 cases; and bone invasive stage, five cases) the number of atypical melanocytes and their degree of nuclear atypia increased, and the ascent of atypical melanocytes and pagetoid spread became conspicuous. Mitoses became apparent only from the progressive stage. From these observations, we would like to propose three new pathological clues of early stage subungual melanoma: (i) "skip lesion", proliferation of the tumor cells are more prominent in the hyponychium than in the nail bed or nail matrix; (ii) histological confirmation of Hutchinson's sign; and (iii) epithelial thickening and/or compact arrangement of the elongated basal cells. PMID- 19120764 TI - Ten cases of sebaceous carcinoma arising in nevus sebaceus. AB - Although nevus sebaceus is known to develop various types of secondary neoplasms, it rarely causes carcinoma and only 14 cases of secondary sebaceous carcinoma have been reported. In this study, 10 cases of sebaceous carcinoma arising in nevus sebaceus were collected. The clinicopathological features and results of immunohistochemical examinations with adipophilin, perilipin and p53 were summarized. Sebaceous carcinoma arising in nevus sebaceous predominantly occurred on the scalp (8/10) of elderly women (mean age, 67.7 years). No case was associated with Muir-Torre syndrome. We found several pathological features of sebaceous carcinoma; that is, made up mainly of germinative cells, moderate nuclear atypia without pleomorphism and many mitoses (4-28/10 high-power field). Adipophilin and perilipin antibodies highlighted lipid drops in the cytoplasm of the malignant cells in all cases. Overexpression of p53 was seen in all cases. In two cases there were coexisting benign-looking sebaceous lesions at the periphery of the main cancer nodule, and in these lesions p53 showed low positivity compared with the clearly malignant area. There was co-occurrence of another neoplasm in three cases with trichoblastoma, sebaceoma and syringocystadenoma papilliferum, respectively. All cases were treated by excision of the malignant lesion, with or without inclusion of the nevus sebaceus. In a follow-up period of 1-7 years, there was no case of recurrence, lymph node metastases or distant metastases. With these specific pathological and immunohistochemical findings using adipophilin, perilipin and p53, we have to consider the possibility that there is a tendency to underdiagnose secondary sebaceous carcinomas in nevus sebaceus. These clinicopathological features of sebaceous carcinomas developing in the nevus sebaceus seem to indicate different biological entities from de novo sebaceous carcinoma. PMID- 19120765 TI - Topical tacrolimus 0.1% ointment in the treatment of localized scleroderma. An open label clinical and histological study. AB - Localized scleroderma or morphea, although a self-limited and benign disease, may leave substantial physical and cosmetic deformity necessitating treatment but treatment remains to date unsatisfactory. The aim of our study was to evaluate the efficacy of topical tacrolimus ointment in the treatment of morphea. Thirteen patients with morphea used tacrolimus 0.1% cream b.i.d. without occlusion for 4 months. Patients were followed up for up to a year. A 4-mm biopsy was taken before starting treatment in seven patients and 4 months after continuous use of tacrolimus 0.1% ointment, next to the previous biopsy site. Masson trichrome and elastica stains were performed to evaluate the distribution of elastic fibers as well as the streptavidin-biotin horseradish peroxide immunohistochemical method for the detection of CD20/L-26, CD3, CD8, CD4, CD1a, human leukocyte antigen-DR and CD25. Four patients had a less than 25% improvement, two patients responded by 50-70% and the remaining seven by more than 70%. Patients with thick, well established lesions responded poorly in comparison to others with less thick and more erythematous ones. Patients with mild-to-moderate fibrosis histologically were more likely to improve after treatment, while the lymphocytic infiltrate decreased regardless of initial degree before treatment. It was concluded that topical tacrolimus 0.1% cream may be used in patients with morphea, particularly with early inflammatory lesions, even as a first-line treatment. PMID- 19120766 TI - Five cases of necrotizing fasciitis: lack of skin inflammatory signs as a clinical clue for the fulminant type. AB - Herein, we describe five patients with necrotizing fasciitis (NF) who had variable outcomes and clinical manifestations. At the onset, all patients exhibited purpura with or without blister and ulceration accompanied by severe pain and tenderness in the affected skin. Out of five patients, three lacked inflammatory signs such as redness and heat, and two of the three patients showed fulminant progression and died despite intensive treatments including surgical debridement, antimicrobial therapy, close monitoring and physiological support. Tissue specimens from the patients without skin inflammatory signs showed mild neutrophil infiltration in addition to necrosis from the epidermis to subcutaneous fat, and variable amounts of thrombi. Furthermore, numerous bacteria were detected by Gram stain. By contrast, the remaining two patients with skin inflammatory signs revealed slower progression, and tissue specimens from both patients showed heavy neutrophil infiltration, but bacteria were hardly detected. Therefore, these cases suggest the possibility that the paucity of skin inflammatory signs, such as redness and heat, in NF may be a clinical clue to predict the fulminant type. PMID- 19120767 TI - Giant microcystic adnexal carcinoma of the scalp. AB - Microcystic adnexal carcinoma (MAC) is an uncommon, locally aggressive tumor. It typically involves the upper lip of middle-aged adults, and in rare instances the scalp. We report a Japanese woman with a giant MAC on the scalp. Physical examination revealed a 110 mm x 120 mm induration on her parietal region. Microscopically, the tumor showed both pilar and sweat gland differentiation. Resection included the cranium; for reconstruction we used a titan mesh allograft and covered it with a free latissimus dorsi muscle flap and a mesh skin graft. Ours is the first case of a MAC measuring more than 100 cm2 arising on the scalp of an individual in the third decade of life. PMID- 19120768 TI - Postmenopausal frontal fibrosing alopecia in a Japanese woman with Sjogren's syndrome. AB - Postmenopausal frontal fibrosing alopecia (PFFA) is a rare alopecia that develops in the frontoparietal scalp of postmenopausal women. Etiology of PFFA is unknown. Most of cases of PFFA have been reported in European and North American countries. Herein, we report a Japanese case of PFFA associated with Sjogren's syndrome. A 66-year-old woman had had slowly progressive, band-like, scarring alopecia on her frontoparietal scalp. Hair follicles on the margin showed follicular keratosis. Histologically, fibrosis and lymphocytic infiltration were mild. This case suggests that PFFA may show mild inflammatory reaction and mild fibrosis in Japanese women. The association with immunological disorders including Sjogren's syndrome should be studied further. PMID- 19120769 TI - Transition of psoriasiform drug eruption to psoriasis de novo evidenced by histopathology. AB - Drug reaction can be one of the triggering factors either for exacerbation of pre existing psoriasis or precipitation of psoriasis de novo. Herein, we report a case with psoriasiform drug eruption due to pravastatin, but it relapsed after discontinuance of the drug. Serial investigation of the histopathology and immunohistochemistry with anti-signal transducer and activator of transcription 3 (STAT3) revealed drug reaction-associated lesions intermingled with psoriasiform changes, which later predominated, suggesting a conversion into psoriasis de novo. PMID- 19120770 TI - Linear IgA dermatosis: report of an infantile case and analysis of 213 cases in Japan. AB - A 3-year-old boy presented with multiple vesicles, showing a rosette-like arrangement around the crusts. Histopathological and immunohistochemical examinations demonstrated subepidermal blistering with neutrophilic infiltration associated with deposition of IgA, but not IgG, linearly distributed along the basement membrane zone (BMZ) of the epidermis. Indirect immunofluorescence revealed circulating antibodies (IgA class, x160) against the BMZ of guinea pig lip skin. Based on the diagnosis of linear IgA dermatosis (LAD) of childhood, administration of dexamethasone (2 mg/day) was started, and the eruptions diminished immediately. Western blot analysis using extract of the HaCaT cell as a substrate, demonstrated the corresponding antigen at 120-kDa molecular weight. There have been 213 cases of LAD reported in Japan including conference abstracts and these were studied to determine whether infantile cases differed from adult ones, and whether cases associated with IgG as well as IgA (IgA/G type), differed from the cases associated with IgA only (IgA type). IgG contributed less frequently to the infantile type (age of onset, < or =15 years) than to the adult type (age of onset, > or =16 years). Clinical appearance did not show any obvious difference between the IgA/G type and IgA type. However, three-quarters of cases showing localization of antigen to the dermal side were the IgA/G type. PMID- 19120771 TI - Use of topical povidone-iodine resulting in an iododerma-like eruption. AB - Iododerma is a rare skin eruption that is usually induced by the systemic use of iodide-containing radiographic contrast medium or treatment with oral potassium iodide therapy. Iododerma has also rarely been reported to occur following topical application of iodine. We herein report the case of a 42-year-old male who developed multiple pinpoint pustules on both lower extremities. Three days after the eruption began, the patient started applying topical 10% povidone iodine solution to the lesions. During this treatment, the lesions enlarged into multiple 2-9-cm pus-filled bullae, limited to the treated areas. Lesional bacterial culture swab and Gram stain were negative. Blood cultures were also negative and the patient's white blood cell count was normal. A punch biopsy specimen revealed pseudoepitheliomatous hyperplasia and dermal-epidermal separation with epidermal necrosis and underlying abscess formation. A superficial and deep perivascular, interstitial and perifollicular infiltrate containing numerous neutrophils, with abscess formation and eosinophils was also seen. This histology was consistent with iododerma and the patient's lesions involuted following cessation of topical iodine use, leaving only post inflammatory hyperpigmentation. In summary, our patient likely had a folliculitis that was then treated with topical povidone-iodine that led to iododerma. Although it remains possible that the reaction was an unusual contact hypersensitivity response, the ulceration, pseudoepitheliomatous hyperplasia and abscess formation do support an iododerma-like eruption. PMID- 19120772 TI - Primary cutaneous anaplastic large cell lymphoma successfully treated with local thermotherapy using pocket hand warmers. AB - Apart from for cutaneous deep fungal or mycobacterial infections, thermotherapy has been used for various malignant tumors. We report a case of primary cutaneous anaplastic large cell lymphoma, which responded quite well to topical thermotherapy using chemical pocket hand warmers. The treatment resulted in an immediate tumor regression without recurrence. This method is simple and might be a useful tool against solitary cutaneous lymphoma, especially of elderly patients with poor performance status or with various systemic complications. PMID- 19120773 TI - Case of cystic schwannoma. AB - Schwannoma is a well-defined tumor arising from the nerve sheath. It may present as a solitary mass in any part of the body, but is more commonly seen in the head, neck and extremities. We describe a 32-year-old female patient with a small dermal nodule on the forehead, which was 2 cm in diameter, round and of a cystic nature. Microscopic examination revealed that it consisted of compact spindle cells arranged partly in short bundles or a fascicular pattern with outstanding cystic degeneration manifested as a large, unilocular cavity in the center of the tumor tissue. The tumor cells were positive for S-100 protein and negative for epithelial membrane antigen (EMA), while the tumor capsule was positive for EMA. There was no S-100-positive membrane-like structure lining the cyst wall. PMID- 19120774 TI - Two cases of subungual melanoma in situ. AB - Melanonychia, which is characterized by brown or black pigmentation within the nail plate, includes heterogeneous conditions such as pigmented nevus, subungual melanoma and lentigo. We treated two cases of subungual melanoma in situ. One case was a 58-year-old woman who suffered from a malignant melanoma in situ of the left third fingernail, who had also suffered from melanonychia of the fingers for more than 30 years. She had a past history of carcinoma of the uterine cervix. The other patient was a 42-year-old man, who suffered from a malignant melanoma in situ of the right fifth fingernail. He had a past history of carcinoma of the stomach for which he had undergone surgery 2 years earlier. Both cases were accompanied by Hutchinson's sign on the fingertip skin, and the presence of this sign led to the correct diagnosis of subungual melanoma in situ. Judging from previously reported cases, it is unlikely that patients with malignant melanoma have an increased risk of carcinoma of the uterine cervix or of the stomach. PMID- 19120775 TI - Occupational generalized argyria after exposure to aerosolized silver. PMID- 19120776 TI - Debridement using a dermatome driven with compressed air (airdermatome). PMID- 19120777 TI - The enamel preservation fee? PMID- 19120778 TI - Clinical performance of low fusing framework veneering ceramic materials. PMID- 19120779 TI - The use of dental implants in combination with removable partial dentures: a case report. AB - Implant treatment protocols for fully edentulous patients include implant supported fixed restorations with a minimum of five to six implants or overdentures on two to four implants. However, implant placement in the posterior areas is sometimes not permitted by anatomic and/or financial restraints, or patients' unwillingness to have extensive surgical procedures. The purpose of this case report is to present a treatment option where bilateral distal extension removable partial dentures (RPDs) are used in combination with anterior fixed implant prostheses with semi-precision attachments. The initial treatment plan of a 65-year-old female with failing dentition involved the fabrication of overdentures supported by four implants placed in the interforaminal area in both arches. Because of inadequate space for the retentive elements of the overdenture and the patient's objection to surgical procedures for the placement of additional implants to support a fixed restoration, the treatment plan was modified. Fixed ceramometal cement-retained implant restorations with semi precision attachments adjacent to the distal abutments were fabricated in the maxilla and the mandible. Bilateral distal extension RPDs were placed in both arches. The patient's benefits were increased comfort, good esthetics in the anterior area, improved phonetics, and masticatory function. Retentive element maintenance requirements were similar to conventional RPDs. More extended controlled clinical studies are needed to establish the long-term success of this treatment option. CLINICAL SIGNIFICANCE: Removable partial dentures may offer an attractive treatment option for an edentulous patient, as they may combine an FPD in the anterior segment with a removable appliance in the posterior areas. A satisfying restoration can be achieved with fewer implants. PMID- 19120781 TI - Soft tissue cone-beam computed tomography: a novel method for the measurement of gingival tissue and the dimensions of the dentogingival unit. AB - The dimensions and relationships of the structures of the dentogingival unit have been greatly overlooked because of the inability to easily and precisely determine them. The purpose of the present study was to develop a soft tissue cone-beam computed tomography (ST-CBCT) to improve soft tissue image quality and allow the determination of the dimensions and relationships of the structures of the dentogingival unit. Two separate CBCT scans were obtained from three patients with different periodontal biotypes. The first was a scan following standard methods; however, for the ST-CBCT the patients wore a plastic lip retractor and retracted their tongues toward the floor of their mouths. With the first scan, only measurements of the distance of the cementoenamel junctional (CEJ) to the facial bone crest, and the width of the facial alveolar bone were possible. In contrast, ST-CBCT allowed measurements of the distance of the gingival margin to the facial bone crest, the gingival margin to the CEJ, and width of the facial gingiva. ST-CBCT scans allowed a clear visualization, measurement of the dimensions, and analysis of the relationship of the structures of the periodontium and dentogingival attachment apparatus. CLINICAL SIGNIFICANCE: The dimensions and relationships of the structures of the dentogingival attachment apparatus are essential aspects in many fields of dentistry and this report describes a simple, novel, and noninvasive technique to determine them. This technique may aid clinicians in the planning and execution of procedures in several dental specialties. PMID- 19120783 TI - Reproducibility of the pink esthetic score--rating soft tissue esthetics around single-implant restorations with regard to dental observer specialization. AB - PURPOSE: The pink esthetic score (PES) evaluates the esthetic outcome of soft tissue around implant-supported single crowns in the anterior zone by awarding seven points for the mesial and distal papilla, soft-tissue level, soft-tissue contour, soft-tissue color, soft-tissue texture, and alveolar process deficiency. The aim of this study was to measure the reproducibility of the PES and assess the influence exerted by the examiner's degree of dental specialization. MATERIALS AND METHODS: Fifteen examiners (three general dentists, three oral maxillofacial surgeons, three orthodontists, three postgraduate students in implant dentistry, and three lay people) applied the PES to 30 implant-supported single restorations twice at an interval of 4 weeks. Using a 0-1-2 scoring system, 0 being the lowest, 2 being the highest value, the maximum achievable PES was 14. At the second assessment, the photographs were scored in reverse order. Differences between the two assessments were evaluated with the Spearman's rank correlation coefficient (R). The Wilcoxon signed-rank test was used for comparisons of differences between the ratings. A significance level of p < 0.05 was chosen for both tests. RESULTS: Observer results indicated that the agreement between the first and second rating for all occupational groups was 70.5%, with a broad correlation between the two ratings and a high statistical significance (Spearman's R = 0.58, p = 0; Wilcoxon T = 163,182, Z = 3.383599, p = 0.000716). The most agreement between the first and second rating was obtained by orthodontists with 73.5% (R = 0.67), and the least by lay people 65.9% (R = 0.50). Very poor and very esthetic restorations showed the smallest deviations. Orthodontists were found to have assigned significantly poorer ratings than any other group. The assessment of postgraduate students and laypersons were the most favorable. CLINICAL SIGNIFICANCE: The PES allows for a more objective appraisal of the esthetic short- and long-term results of various surgical and prosthetic implant procedures. It reproducibly evaluates the peri-implant soft tissue around single-implant restorations and results in good intra-examiner agreement. However, an effect of observer specialization on rating soft-tissue esthetics can be shown. PMID- 19120785 TI - Shear bond strength of a sealant to contaminated-enamel surface: influence of erbium : yttrium-aluminum-garnet laser pretreatment. AB - BACKGROUND: Salivary contamination is one of the factors that can disturb the sealing process and interfere in the longevity of pit and fissure sealants. Erbium : yttrium-aluminum-garnet (Er : YAG) laser could influence the bond strength of enamel and increase the acid resistance. PURPOSE: To evaluate the influence of Er : YAG laser on the shear bond strength of a sealant to a salivary contaminated enamel surface. METHODS: Twenty-four third molars had the roots sectioned 2 mm coronal to the cementoenamel junction. The crowns were mesiodistally sectioned providing 48 halves that were embedded in polyester resin. Enamel was flattened and a 2-mm diameter bonding area was demarcated. Specimens were randomly assigned to two groups according to the superficial pretreatment-37% phosphoric acid (A) and Er : YAG laser (80 mJ/2 Hz) + phosphoric acid (L), which were subdivided into two groups (N = 12), without salivary contamination (C) and with salivary contamination (SC). To contaminate the specimens, 0.25 mL of human fresh saliva was applied for 20 seconds and then dried. Fluroshield sealant was applied in all specimens. After storage, shear bond strength of samples were tested in a universal testing machine. RESULTS: Means in MPa were: AC-14.61 (+/-2.52); ASC-6.66 (+/-2.34); LC-11.91 (+/-1.34); and LSC-2.22 (+/-0.66). Statistical analysis revealed that surfaces without salivary contamination and with acid treatment had the highest mean (p < 0.05). The group with salivary contamination treated by Er : YAG laser followed by phosphoric acid application presented the lowest bond values (p < 0.05). CONCLUSIONS: The phosphoric acid etching under dry condition yielded better bonding performance. Er : YAG laser was not able to increase the effectiveness of conventional acid etching of enamel in the bond of sealants in both dry and wet conditions. CLINICAL SIGNIFICANCE: Under the conditions of this study, the conventional etching protocol (phosphoric acid without salivary contamination) is still preferable to laser-conditioning enamel surface prior to sealant application. PMID- 19120787 TI - Bleaching agents with varying concentrations of carbamide and/or hydrogen peroxides: effect on dental microhardness and roughness. AB - PURPOSE: To evaluate the effect of low and highly concentrated bleaching agents on microhardness and surface roughness of bovine enamel and root dentin. METHODS: According to a randomized complete block design, 100 specimens of each substrate were assigned into five groups to be treated with bleaching agents containing carbamide peroxide (CP) at 10% (CP10); hydrogen peroxide (HP) at 7.5% (HP7.5) or 38% (HP38), or the combination of 18% of HP and 22% of CP (HP18/CP22), for 3 weeks. The control group was left untreated. Specimens were immersed in artificial saliva between bleaching treatments. Knoop surface microhardness (SMH) and average surface roughness (Ra) were measured at baseline and post-bleaching conditions. RESULTS: For enamel, there were differences between bleaching treatments for both SMH and Ra measurements (p = 0.4009 and p = 0.7650, respectively). SMH significantly increased (p < 0.0001), whereas Ra decreased (p = 0.0207) from baseline to post-bleaching condition. For root dentin, the group treated with CP10 exhibited the significantly highest SMH value differing from those groups bleached with HP18/CP22, HP7.5, which did not differ from each other. Application of HP38 resulted in intermediate SMH values. No significant differences were found for Ra (p = 0.5975). Comparing the baseline and post bleaching conditions, a decrease was observed in SMH (p < 0.0001) and an increase in Ra (p = 0.0063). CONCLUSION: Bleaching agents with varying concentrations of CP and/or HP are capable of causing mineral loss in root dentin. Enamel does not perform in such bleaching agent-dependent fashion when one considers either hardness or surface roughness evaluations. CLINICAL SIGNIFICANCE Bleaching did not alter the enamel microhardness and surface roughness, but in root dentin, microhardness seems to be dependent on the bleaching agent used. PMID- 19120789 TI - Critical appraisal: Protocol for the prevention and management of root caries. AB - This critical appraisal deviates from our standard format to address a timely topic of particular interest to clinicians. PMID- 19120790 TI - Occlusion (part 2 of 2). AB - In the previous Journal of Esthetic and Restorative Dentistry's Talking with Patients, we defined occlusion and malocclusion and discussed when and why malocclusion is a concern. In this issue we will discuss the diagnosis and treatment of malocclusions. PMID- 19120791 TI - The history of Toxoplasma gondii--the first 100 years. AB - In this paper the history of Toxoplasma gondii and toxoplasmosis is reviewed. This protozoan parasite was first discovered in 1908 and named a year later. Its medical importance remained unknown until 1939 when T. gondii was identified in tissues of a congenitally infected infant, and veterinary importance became known when it was found to cause abortion storms in sheep in 1957. The discovery of a T. gondii specific antibody test, Sabin-Feldman dye test in 1948 led to the recognition that T. gondii is a common parasite of warm-blooded hosts with a worldwide distribution. Its life cycle was not discovered until 1970 when it was found that felids are its definitive host and an environmentally resistant stage (oocyst) is excreted in feces of infected cats. The recent discovery of its common infection in certain marine wildlife (sea otters) indicates contamination of our seas with T. gondii oocysts washed from land. Hygiene remains the best preventive measure because currently there is no vaccine to prevent toxoplasmosis in humans. PMID- 19120792 TI - How epigenomics contributes to the understanding of gene regulation in Toxoplasma gondii. AB - How apicomplexan parasites regulate their gene expression is poorly understood. The complex life cycle of these parasites implies tight control of gene expression to orchestrate the appropriate expression pattern at the right moment. Recently, several studies have demonstrated the role of epigenetic mechanisms for control of coordinated expression of genes. In this review, we discuss the contribution of epigenomics to the understanding of gene regulation in Toxoplasma gondii. Studying the distribution of modified histones on the genome links chromatin modifications to gene expression or gene repression. In particular, coincident trimethylated lysine 4 on histone H3 (H3K4me3), acetylated lysine 9 on histone H3 (H3K9ac), and acetylated histone H4 (H4ac) mark promoters of actively transcribed genes. However, the presence of these modified histones at some non expressed genes and other histone modifications at only a subset of active promoters implies the presence of other layers of regulation of chromatin structure in T. gondii. Epigenomics analysis provides a powerful tool to characterize the activation state of genomic loci of T. gondii and possibly of other Apicomplexa including Plasmodium or Cryptosporidium. Further, integration of epigenetic data with expression data and other genome-wide datasets facilitates refinement of genome annotation based upon experimental data. PMID- 19120793 TI - Calmodulin localization and its effects on endocytic and phagocytic membrane trafficking in Paramecium multimicronucleatum. AB - In ciliates, calmodulin (CaM), as in other cells, has multiple functions, such as activation of regulatory enzymes and modulating calcium-dependent cellular processes. By immunogold localization, CaM is concentrated at multiple sites in Paramecium. It is seen scattered over the cytosol, but bound to its matrix, and is concentrated at the pores of the contractile vacuole complexes and with at least three microtubular arrays. It was localized peripheral to the nine-doublet microtubules of the ciliary axonemes. The most striking localization was on the akinetic side only of the cytopharyngeal microtubular ribbons opposite the side where the discoidal vesicles, acidosomes and the 100-nm carrier vesicles bind and move. CaM was also present at the periphery of the postoral microtubular bundles along which the early vacuole moves and was associated with the cytoproct microtubules that guide the spent digestive vacuoles to the cytoproct. It was not found on the membranes of, or in the interior of nuclei, mitochondria, phagosomes, and trichocysts, and was only sparsely scattered over the cytosolic sides of discoidal vesicles, acidosomes, lysosomes, and digestive vacuoles. Together the associations with specific microtubular arrays and the effects of trifluoperazine and calmidazolium indicate that CaM is involved (i) in vesicle transport to the cytopharynx area for vacuole formation and subsequent vacuole acidification, (ii) in early vacuole transport along the postoral fiber, and (iii) in transporting the spent vacuole to the cytoproct. Higher CaM concentrations subjacent to the cell's pellicle and close to the decorated tubules of the contractile vacuole complex may support a role for CaM in ion traffic. PMID- 19120794 TI - Trichopodiella faurei n. sp. (Ciliophora, Phyllopharyngea, Cyrtophoria): morphological description and phylogenetic analyses based on SSU rRNA and group I intron sequences. AB - A new marine cyrtophorian ciliate Trichopodiella faurei n. sp., which belongs to the order Dysteriida, family Hartmannulidae, was investigated at the morphological and molecular levels. A combination of morphological features of the organism including the oval body shape, 2-3 contractile vacuoles, 22-28 nematodesmal rods in the cytopharyngeal basket, and 31-39 somatic kineties, distinguishes it from all other known congeners. In reconstructed small subunit (SSU) rRNA phylogenies, T. faurei groups with Isochona, a representative genus of the subclass Chonotrichia. The similarity of the infraciliature between hartmannulids and several chonotrichian examples also suggests that these taxa should be closely related. A new S943 intron belonging to group IC1 was identified in the SSU rRNA gene of this species. This intron is phylogenetically related to the S891 introns previously found in the suctorians Acineta sp. and Tokophrya lemnarum, and their internal guide sequences share four nucleotides, indicating that these introns were vertically inherited from a common phyllopharyngean ancestor and that reverse splicing might have been involved in the transposition. PMID- 19120795 TI - The zooflagellates Stephanopogon and Percolomonas are a clade (class Percolatea: Phylum Percolozoa). AB - The enigmatic marine protozoan Stephanopogon was first classified with ciliate protozoa because its pellicle also has rows of cilia. As ciliates have nuclear dimorphism with separate germline and somatic nuclei, Stephanopogon with several identical nuclei was regarded as a model for a hypothetical homokaryotic ancestor of ciliates. When electron microscopy revealed radical differences from ciliates this idea was abandoned, but its evolutionary position remains controversial, affinities with three other phyla being suggested. We sequenced 18S rDNA from Stephanopogon aff. minuta and actin genes from it and Stephanopogon apogon to clarify their evolutionary position. Phylogenetic analyses of 18S rRNA nest S. aff. minuta and Stephanopogon minuta securely within the protozoan phylum Percolozoa with zooflagellates of the genus Percolomonas, their closest relatives, comprising the clade Percolatea. This supports a previous grouping of Stephanopogon (order Pseudociliatida) with Percolomonas (order Percolomonadida) as a purely zooflagellate class Percolatea within Percolozoa, in contrast to the fundamentally amoeboid Heterolobosea, which are probably ancestral to Percolatea. Stephanopogon actins evolve exceptionally fast: actin trees place them as a long branch within bikont eukaryotes without revealing their sisters. We establish Percolomonadidae fam. n. for Percolomonas, excluding Pharyngomonas kirbyi g., sp. n. and Pharyngomonas (=Tetramastix=Percolomonas) salina comb. n., which unlike Percolomonas have two anterior and two posterior cilia and a pocket-like pharynx, like "Macropharyngomonas", now grouped with Pharyngomonas as a new purely zooflagellate class Pharyngomonadea, within a new subphylum Pharyngomonada; this contrasts them with the revised ancestrally amoeboflagellate subphylum Tetramitia. We discuss evolution of the percolozoan cytoskeleton and different body forms. PMID- 19120796 TI - Morphology and small subunit rDNA gene sequence of Pseudoamphisiella quadrinucleata n. sp. (Ciliophora, Urostylida) from the South China Sea. AB - The urosylid genus Pseudoamphisiella was established by Song (1996) with hitherto only two known congeners. In the present work, the morphology and infraciliature of a new member, Pseudoamphisiella quadrinucleata n. sp., a form with conspicuous alveolar layer and four macronuclear nodules isolated from the coastal waters both near Hong Kong and near Guangzhou, South China were investigated using living observation and protargol silver impregnation methods. Pseudoamphisiella quadrinucleata differs from other two known forms mainly by the number of macronuclear nodules: constantly four vs. two in Pseudoamphisiella alveolata and 24-57 in Pseudoamphisiella lacazei. To support this, the sequence of the small subunit rDNA of P. quadrinucleata n. sp. showed 14 and 74 nucleotides in comparison with that of the two known congeners, respectively, which hence firmly supports the validity of the new species. PMID- 19120797 TI - Micronucleus-specific bacterium Holospora elegans irreversibly enhances stress gene expression of the host Paramecium caudatum. AB - The bacterium Holospora is an endonuclear symbiont of the ciliate Paramecium. Previously, we reported that paramecia bearing the macronuclear-specific symbiont Holospora obtusa survived better than symbiont-free paramecia, even under high temperatures unsuitable for growth. The paramecia with symbionts expressed high levels of hsp70 mRNAs even at 25 degrees C, a usual growth temperature. We report herein that paramecia bearing the micronuclear-specific symbiont Holospora elegans also acquire the heat-shock resistance. Even after the removal of the bacteria from the hosts by treatment with penicillin, the resulting aposymbiotic paramecia nevertheless maintained their heat shock-resistant nature for over 1 yr. Like symbiotic paramecia, these aposymbiotic paramecia also expressed high levels of both hsp60 and hsp70 mRNAs even at 25 degrees C. Moreover, analysis by fluorescent in situ hybridization with a probe specific for Holospora 16S rRNA revealed that the 16S rRNA of H. elegans was expressed around the nucleoli of the macronucleus in the aposymbiotic cells. This result suggests the possible transfer of Holospora genomic DNA from the micronucleus into the macronucleus in symbiotic paramecia. Perhaps this exogenous DNA could trigger the aposymbiotic paramecia to induce a stress response, inducing higher expression of Hsp60 and Hsp70, and thus conferring heat-shock resistance. PMID- 19120798 TI - Molecular characterization of two types of rDNA units in a single strain of Candida albicans. AB - Strains of the opportunistic fungal pathogen Candida albicans vary in the presence or absence of a self-splicing group I intron ribozyme (Ca.LSU) in the 25S rRNA gene on chromosome R. Strains of C. albicans typically either lack or contain this ribozyme. However, some strains have both intron-containing and intronless rRNA genes (rDNA). Pulsed-field gel electrophoresis analysis of undigested and restricted DNA showed at least six different karyotypes among eight independent colonies of such a heteroallelic strain. In each case, the variation was in chromosome R, and was due to changes in the number of rDNA units. In strains with only one type of rDNA, chromosome R also varied considerably. Polymerase chain reaction amplification spanning the rDNA unit demonstrated that intron-containing rDNA units are tandemly arrayed, and are immediately adjacent to intronless units in the same cluster. Both types of units were present in the rDNA clusters of both R chromosomes. Possible explanations of these results are loss of Ca.LSU group I intron through purifying selection and/or a relaxation of the commonly accepted concerted evolution of the rDNA units. PMID- 19120799 TI - Nuclear genome sequence survey of the Dinoflagellate Heterocapsa triquetra. AB - Dinoflagellates have among the largest nuclear genomes known, but we know little about their contents or organisation. Given the interest in dinoflagellate ecology, cell biology, and evolutionary biology, there are many reasons to thoroughly investigate the contents of dinoflagellate genomes, but because of their large size the only thorough samples to date have relied on expressed sequence tag surveys to analyse cDNAs. To complement this, there are some studies of the physical properties of dinoflagellate chromosomes, but no direct survey of the nature of the sequences contained within them. To start to build a picture of the contents of these genomes, we have sequenced over 230,000 bp from the nuclear genome of Heterocapsa triquetra, which has been estimated to be 18-23 billion base pairs in total. The survey includes one putative gene with two relict spliced leaders, one putative pseudogene, and a small number of low-complexity repeats, transposons, and other putative selfish elements, all of which account for about 5% of the survey. Another 5% of the survey was long, complex repeats, some highly represented. By far the greatest fraction of the survey (89.5%) is made up of non-repeated sequence with no similarity to any other known sequence. PMID- 19120800 TI - A Sarcocystid misidentified as Hepatozoon didelphydis: molecular data from a parasitic infection in the blood of the Southern mouse opossum (Thylamys elegans) from Chile. AB - The blood of 21 adult South American mouse opossums (Thylamys elegans) captured from April through August of 2005 in central Chile was examined for parasites. Light microscopic analysis of blood smears initially suggested that a highly pleomorphic Hepatozoon species typical of American opossums was infecting erythrocytes. Unexpectedly, amplification by PCR and sequencing of a DNA fragment of the small subunit rDNA combined with phylogenetic analyses indicated that the parasite is not a member of the suborder Adeleorina, which includes the Haemogregarina and Hepatozoon species, but that it is a clearly distinct member of the suborder Eimeriorina, which includes the cyst-forming family Sarcocystidae. Therefore, a reclassification of this unusual intraerythrocytic apicomplexan will require additional life cycle, microscopic, and molecular analyses. PMID- 19120801 TI - The unique adaptation of the life cycle of the coelomic gregarine Diplauxis hatti to its host Perinereis cultrifera (Annelida, Polychaeta): an experimental and ultrastructural study. AB - The coelomic gregarine Diplauxis hatti exhibits a unique adaptation of its life cycle to its polychaete host Perinereis cultrifera. Experimental and ultrastructural observations on natural populations from the English Channel showed that release of parasite spores is concomitant with the polychaete spawning. As the development of P. cultrifera is direct, the notochete larva ingest parts of the jelly coat covered with numerous sporocysts of D. hatti during hatching. Transepithelial migration of the sporozoites takes place in the gut of three- or four-segment notochete larvae and syzygies of about 20 microm are observed in the coelom. Growth of these young syzygies is slow: after 18-24 mo they reach only 60-70 microm. They exhibit active pendular movements. In the English Channel, female and male gametogenesis of P. cultrifera begins at 19 mo and 2 yr, respectively; the somatic transformations (epitoky) in the last 4 mo of their 3-year life. During epitoky, the syzygies undergo an impressive growth and reach 700-800 microm within a few weeks. A shift from pendular to active peristaltic motility is observed when the syzygies reach 200-250 microm. When gamogony occurs, syncytial nuclear divisions are initiated and cellularization produces hundred to thousands of male and female gametes of similar size. The male gametes exhibit a flagellum with 3+0 axoneme. The mixing of the gametes ("danse des gametes") and fertilization are observed during 4-5 h. Zygotes differentiate sporoblasts with eight sporozoites. The sporozoites exhibit the canonical structure of Apicomplexa, a polarized cell with micronemes and rhoptries. PMID- 19120802 TI - Gene cloning and biochemical characterization of an alcohol dehydrogenase from Euglena gracilis. AB - Euglena gracilis is a freshwater free-living organism able to grow with ethanol as carbon source; to facilitate this metabolism several alcohol dehydrogenase (ADH) activities have been detected. We report the gene cloning, over-expression, and biochemical characterization of a medium-chain NAD(+)-dependent ADH from E. gracilis (EgADH). The enzyme's amino acid sequence displayed the highest percentages of similarity and identity with ADHs of bacteria and fungi. In the predicted three-dimensional model, all the residues involved in Zn(2+), cofactor, and substrate binding were conserved. A conventional signal peptide for import into mitochondria could not be clearly identified. The protein of 37 kDa was over expressed, purified to homogeneity, and kinetically characterized. The enzyme's optimal pH was 7.0 for ethanol oxidation displaying a V(m) of 11.7+/-3.6 U/mg protein and a K(m) of 3.2+/-0.7 mM for this substrate. Isopropanol and isopentanol were also utilized, although with less efficiency. It showed specificity for NAD(+) with a K(m) value of 0.39+/-0.1 mM and Mg(2+) or Zn(2+) were essential for activity. The recombinant EgADH reported here may help to elucidate the roles that different ADHs have on the metabolism of short- and long chain alcohols in this microorganism. PMID- 19120803 TI - Phylogeny of six genera of the subclass Haptoria (Ciliophora, Litostomatea) inferred from sequences of the gene coding for small subunit ribosomal RNA. AB - The small subunit ribosomal RNA genes of nine species belonging to six genera of litostome ciliates, namely Amphileptus aeschtae, Chaenea teres, Chaenea vorax, Lacrymaria marina, Litonotus paracygnus, Loxophyllum sp.-GD-070419, Loxophyllum jini, Loxophyllum rostratum, and Phialina salinarum, were sequenced for the first time. Phylogenetic trees were constructed using different methods to assess the inter- and intra-generic relationships of haptorians, of which Chaenea, Lacrymaria, Litonotus, and Phialina were analyzed for the first time based on molecular data. Monophyly of the order Pleurostomatida was strongly confirmed, and the two existing families of pleurostomatids, created on the basis of morphology, were confirmed by molecular evidence. Within the Pleurostomatida, Siroloxophyllum utriculariae occupied a well-supported position basal to the Loxophyllum clade, supporting the separation of these genera from one another. Both the subclass Haptoria and the order Haptorida were partially unresolved, possibly paraphyletic assemblages of taxa in all analyses, creating doubts about the traditional placement of some haptorid taxa. The existing sequence of L. rostratum in GenBank (DQ411864) was conspicuously different from that of the isolate from Qingdao, China sequenced in the present work, indicating that they are different species. The isolate from Qingdao was verified as L. rostratum by morphological analysis, and the published morphology of existing GenBank record of L. rostratum is different from it. Based on both morphological and molecular evidence, the latter may be congeneric with an undescribed species of Loxophyllum from Guangdong Province, China. PMID- 19120804 TI - Parastrombidinopsis minima n. sp. (Ciliophora: Oligotrichia) from the coastal waters of northeastern Taiwan: morphology and small subunit ribosomal DNA sequence. AB - Parastrombidinopsis minima n. sp. is investigated, using live observations, protargol preparations, and molecular data. In living cells, the ranges of cell length are 85-95 microm, cell width 60-70 microm, and oral diameter 40-50 microm. In protargol-impregnated specimens, cell length ranges between 43 and 71 microm, cell width between 23 and 42 microm, and oral diameter between 13 and 24 microm. The numbers of external oral polykinetids are 12-16 and of somatic kineties are 11-13. There are always two ovoid macronuclei (9-16 x 4-9 microm). Based on the analysis of morphologic data, the new species can be placed in the family Strombidinopsidae, but based on the small subunit rRNA gene sequence data, the Parastrombidinopsis species are more closely associated with strobilidiids and tintinnids. PMID- 19120807 TI - Island flap employment in the cases of eye outer canthus skin loss. PMID- 19120806 TI - Clinical features and natural history of acquired cold urticaria in a tertiary referral hospital: a 10-year prospective study. AB - BACKGROUND: Acquired cold urticaria (ACU) represents a heterogeneous group of disorders that share a common clinical feature: the development of urticaria or angioedema after cold exposure. We present epidemiological and clinical data of subjects with ACU, natural progression and we examine possible parameters that could correlate with disease severity. METHODS: During a 10-year period in all subjects with ACU, detailed record of personal history, laboratory testing, cold stimulation testing (CST), atopy assessment and disease severity took place. In a re-evaluation visit at the end of the surveillance period, ACU progression was assessed from patients in a subjective way. RESULTS: Four thousand one hundred fifty-seven individuals with chronic urticaria were referred, and 352 (198 males, 154 females, 8.47% of patients with chronic urticaria) presented definite symptoms of physical urticarias, while 95 individuals (49 males, 46 females, 27% of patients with physical urticarias) were detected with ACU. Sixty-two participants were included in study analysis. Thirty-two patients (51.6%) were female; the mean age was 41.5 +/- 15.6 years, while the mean age at disease onset was 32.5 +/- 15.6 years; half were < or = 30 years old at disease onset. The mean duration of surveillance was 9.0 +/- 6.9 years. During this time interval, 18 patients (29.0%) showed the same or even worse symptomatology, 26 patients reported some improvement (41.9%), while in 18 patients, symptoms resolved completely (29.0%); the mean time to resolution was 5.6 +/- 3.5 years. Disease severity was the only variable statistically significantly related to disease progression (P = 0.004). CONCLUSIONS: Cold urticaria is a chronic persistent disorder with occasional severe clinical manifestations. PMID- 19120809 TI - What do human economies, large islands and forest fragments reveal about the factors limiting ecosystem evolution? AB - What factors limit ecosystem evolution? Like human economies, ecosystems are arenas where agents compete for locally limiting resources. Like economies, but unlike genes, ecosystems are not units of selection. In both economies and ecosystems, productivity, diversity of occupations or species and intensity of competition presuppose interdependence among many different agents. In both, competitive dominants need abundant, varied resources, and many agents' products or services, to support the activity and responsiveness needed to maintain dominance. Comparing different-sized land masses suggests that productivity is lower on islands whose area is too small to maintain some of the interdependences that maintain diversity, productivity and competitiveness in mainland ecosystems. Islands lacking the rare, metabolically active dominants that make competition so intense in mainland ecosystems are more easily invaded by introduced exotics. Studies of islets in reservoirs identify mechanisms generating these phenomena. These phenomena suggest how continued fragmentation will affect future 'natural' ecosystems. PMID- 19120810 TI - Male-male competition, female mate choice and their interaction: determining total sexual selection. AB - Empirical studies of sexual selection typically focus on one of the two mechanisms of sexual selection without integrating these into a description of total sexual selection, or study total sexual selection without quantifying the contributions of all of the mechanisms of sexual selection. However, this can provide an incomplete or misleading view of how sexually selected traits evolve if the mechanisms of sexual selection are opposing or differ in form. Here, we take a two-fold approach to advocate a direction for future studies of sexual selection. We first show how a quantitative partitioning and examination of sexual selection mechanisms can inform by identifying illustrative studies that describe both male-male competition and female mate choice acting on the same trait. In our sample, the most common trait where this occurred was body size, and selection was typically linear. We found that male-male competition and female mate choice can be reinforcing or opposing, although the former is most common in the literature. The mechanisms of sexual selection can occur simultaneously or sequentially, and we found they were more likely to be opposing when the mechanisms operated sequentially. The degree and timing that these mechanisms interact have important implications for the operation of sexual selection and needs to be considered in designing studies. Our examples highlight where empirical data are needed. We especially lack standardized measures of the form and strength of selection imposed by each mechanism of sexual selection and how they combine to determine total sexual selection. Secondly, using quantitative genetic principles, we outline how the selection imposed by individual mechanisms can be measured and combined to estimate the total strength and form of sexual selection. We discuss the evolutionary consequences of combining the mechanisms of sexual selection and interpreting total sexual selection. We suggest how this approach may result in empirical progress in the field of sexual selection. PMID- 19120811 TI - Testing multiple hypotheses for the maintenance of male homosexual copulatory behaviour in flour beetles. AB - Diverse animal groups exhibit homosexual interactions, yet the evolutionary maintenance of such behaviours remains enigmatic as they do not directly increase reproductive success by generating progeny. Here, we use Tribolium castaneum flour beetles, which exhibit frequent male homosexual copulations, to empirically test several hypotheses for the maintenance of such behaviours: (1) establishing social dominance; (2) practice for future heterosexual encounters; and (3) indirect sperm translocation. We found no evidence that Tribolium males use homosexual copulations either to establish dominance or to practice behaviours that increase their subsequent heterosexual reproductive performance. Our results provide limited support for the hypothesis of indirect sperm translocation: when males from two genetic strains mated with females immediately following a homosexual copulation, females produced progeny sired not only by the directly mating male, but also by that male's homosexual partner. However, this phenomenon was detected in only 7% of homosexual pairs, and in each case such indirectly sired progeny accounted for < 0.5% of females' total progeny. Direct observations indicated that mounting males often released spermatophores during homosexual copulations. These observations suggest that homosexual copulations may be a behavioural mechanism that allows males to expel older, potentially low-quality sperm. Additional work is needed to test this new hypothesis, and to determine whether sperm release during homosexual copulations occurs in other groups. PMID- 19120812 TI - Overdominant maintenance of diversity in the sea star Pisaster ochraceus. AB - When individuals have higher evolutionary fitness because of being heterozygous at a given gene region, it is known as overdominance. Although overdominant selection could represent an important mechanism for maintaining genetic variation within species, the prevalence of this mode of selection appears to be relatively low. Identification of cases of true single-locus heterozygote advantage are thus useful reference points in our overall understanding of how various forms of balancing selection influence and maintain genetic variation in natural populations. Here we report the apparent long-term maintenance of diversity via overdominant selection with homozygous lethality at an elongation factor locus in the sea star Pisaster ochraceus. Observing this pattern in a gene with such major effects on protein assembly indicates that overdominant selection could be a more prevalent factor in maintaining allelic diversity in the wild than previously recognized. PMID- 19120813 TI - Experimental evidence that competition and habitat use shape the individual fitness surface. AB - A key prediction made by theories of density-dependent competition is that resource overlap should increase the intensity of competition. By extension, we can predict that competition should lead to density-dependent natural selection. I studied natural selection on limb length and body size in a total of seven populations of Anolis sagrei over 3 years in the Bahamas. Experimental manipulations of population density on small off-shore cays revealed that the strength of natural selection on body size increased with density, suggesting that density-dependent intraspecific competition drives natural selection. At low density, reduced competition revealed significant selection on limb length driven by changes in perch diameter, indicating that selection favoured a match between morphology and habitat. The role habitat played in shaping selection was further illuminated by inter-annual changes in vegetation structure stemming from variation in precipitation among years. Thus, changes in both the intensity of competition across spatial replicates, and in resource availability through time, revealed changes in the targets of natural selection. Results provide empirical support for the long-standing hypothesis that density-dependent natural selection shapes the fitness surface of Greater Antilles anoles. PMID- 19120814 TI - Demographic factors and genetic variation influence population persistence under environmental change. AB - Population persistence has been studied in a conservation context to predict the fate of small or declining populations. Persistence models have explored effects on extinction of random demographic and environmental fluctuations, but in the face of directional environmental change they should also integrate factors affecting whether a population can adapt. Here, we examine the population-size dependence of demographic and genetic factors and their likely contributions to extinction time under scenarios of environmental change. Parameter estimates were derived from experimental populations of the rainforest species, Drosophila birchii, held in the lab for 10 generations at census sizes of 20, 100 and 1000, and later exposed to five generations of heat-knockdown selection. Under a model of directional change in the thermal environment, rapid extinction of populations of size 20 was caused by a combination of low growth rate (r) and high stochasticity in r. Populations of 100 had significantly higher reproductive output, lower stochasticity in r and more additive genetic variance (V(A)) than populations of 20, but they were predicted to persist less well than the largest size class. Even populations of 1000 persisted only a few hundred generations under realistic estimates of environmental change because of low V(A) for heat knockdown resistance. The experimental results document population-size dependence of demographic and adaptability factors. The simulations illustrate a threshold influence of demographic factors on population persistence, while genetic variance has a more elastic impact on persistence under environmental change. PMID- 19120815 TI - Evaluation of offspring size-number invariants in 12 species of lizard. AB - The optimal division of resources into offspring size vs. number is one of the classic problems in life-history evolution. Importantly, models that take into account the discrete nature of resource division at low clutch sizes suggest that the variance in offspring size should decline with increasing clutch size according to an invariant relationship. We tested this prediction in 12 species of lizard with small clutch sizes. Contrary to expectations, not all species showed a negative relationship between variance in offspring size and clutch size, and the pattern significantly deviated from quantitative predictions in five of the 12 species. We suggest that the main limitation of current size number models for small clutch sizes is that they rely on assumptions of hierarchical allocation strategies with independence between allocation decisions. Indeed, selection may favour alternative mechanisms of reproductive allocation that avoid suboptimal allocation imposed by the indivisible fraction at low clutch sizes. PMID- 19120816 TI - Metapopulation persistence in fragmented landscapes: significant interactions between genetic and demographic processes. AB - We formulated a mathematical model in order to study the joint influence of demographic and genetic processes on metapopulation viability. Moreover, we explored the influence of habitat structure, matrix quality and disturbance on the interplay of these processes. We showed that the conditions that allow metapopulation persistence under the synergistic action of genetic and demographic processes depart significantly from predictions based on a mere superposition of the effects of each process separately. Moreover, an optimal dispersal rate exists that maximizes the range of survival rates of dispersers under which metapopulation persists and at the same time allows the largest sustainable patch removal and patch-size reduction. The relative impact of patch removal and patch-size reduction depends both on matrix quality and the dispersal strategy of the species: metapopulation persistence is more affected by patch size reduction (patch removal) for low (high)-dispersing species, in presence of a low (high) quality matrix. Avoidance of inbreeding, through increased dispersal when the rate of inbreeding in a population is large, has positive effects on low dispersing species, but impairs the persistence of high-dispersing species. Finally, size heterogeneity between patches largely influences metapopulation dynamics; the presence of large patches, even at the expense of other patches being smaller, can have positive effects on persistence in particular for species of low dispersing ability. PMID- 19120817 TI - HSP70 expression in the Copper butterfly Lycaena tityrus across altitudes and temperatures. AB - The ability to express heat-shock proteins (HSP) under thermal stress is an essential mechanism for ectotherms to cope with unfavourable conditions. In this study, we investigate if Copper butterflies originating from different altitudes and/or being exposed to different rearing and induction temperatures show differences in HSP70 expression. HSP70 expression increased substantially at the higher rearing temperature in low-altitude butterflies, which might represent an adaptation to occasionally occurring heat spells. On the other hand, high altitude butterflies showed much less plasticity in response to rearing temperatures, and overall seem to rely more on genetically fixed thermal stress resistance. Whether the latter indicates a higher vulnerability of high-altitude populations to global warming needs further investigation. HSP70 expression increased with both colder and warmer induction temperatures. PMID- 19120818 TI - Testing evolutionary stasis and trends in first lower molar shape of extinct Italian populations of Terricola savii (Arvicolidae, Rodentia) by means of geometric morphometrics. AB - Extinct populations of Terricola savii have been investigated in order to analyse evolutionary stasis and correlation of first lower molar shape with climatic proxies by means of geometric morphometrics. Evolutionary stasis, its recognition and explanation are central topics in evolutionary paleobiology. In this study, tooth shape variation of the arvicolid T. savii has been analysed through time. In addition to explicit multivariate tests of stasis based on landmark and semi landmark geometric morphometrics, first lower molar M(1) shape has been decomposed in orthogonal axes of variation and tested for correlation with climate changes. Multivariate tests were consistent with evolutionary stasis. Yet, according to univariate tests, the dominant dimension of shape variation shows a temporal trend well correlated with a climatic proxy, i.e. delta(18)O. The remaining variation does not show any trend. Adaptation to current climatic condition might occur even without affecting shape as a whole. Phenotypic plasticity of this species could be invoked to explain evolutionary stasis, as a long time pattern. PMID- 19120819 TI - The evolution of sex-determining mechanisms: lessons from temperature-sensitive mutations in sex determination genes in Caenorhabditis elegans. AB - Sexual reproduction is one of the most taxonomically conserved traits, yet sex determining mechanisms (SDMs) are quite diverse. For instance, there are numerous forms of environmental sex determination (ESD), in which an organism's sex is determined not by genotype, but by environmental factors during development. Important questions remain regarding transitions between SDMs, in part because the organisms exhibiting unique mechanisms often make difficult study organisms. One potential solution is to utilize mutant strains in model organisms better suited to answering these questions. We have characterized two such strains of the model nematode Caenorhabditis elegans. These strains harbour temperature sensitive mutations in key sex-determining genes. We show that they display a sex ratio reaction norm in response to rearing temperature similar to other organisms with ESD. Next, we show that these mutations also cause deleterious pleiotropic effects on overall fitness. Finally, we show that these mutations are fundamentally different at the genetic sequence level. These strains will be a useful complement to naturally occurring taxa with ESD in future research examining the molecular basis of and the selective forces driving evolutionary transitions between sex determination mechanisms. PMID- 19120820 TI - Decompositions of Price's formula in an inhomogeneous population structure. AB - The central tool for the study of allele frequency change due to selection is the remarkably simple but powerful formula of Price [Nature 227 (1970) 520]. Here, I provide what might be called a structural analysis of this formula. The formula essentially accumulates the average allele frequency change over many instances of a fitness-determining interaction, but there are different ways of organizing this average and these lead to quite different computational algorithms. I present three of these: an analysis by population state, an analysis by recipient and an analysis by actor. A comparison of these can lead to a heightened understanding of the different factors behind selective allele frequency change. In particular, I pay attention to the effects of structural inhomogeneity on reproductive value (RV) and emphasize that Price's formula measures RV-weighted allele frequency change. I examine in detail a simple example as a crucial way of cementing the different theoretical pathways. My aim was to produce a simple transparent presentation and therefore I work with a simple population structure and have omitted a number of technical details that are found elsewhere. PMID- 19120821 TI - A sex-linked SCAR marker in Bryonia dioica (Cucurbitaceae), a dioecious species with XY sex-determination and homomorphic sex chromosomes. AB - Genetic crosses between the dioecious Bryonia dioica (Cucurbitaceae) and the monoecious B. alba in 1903 provided the first clear evidence for Mendelian inheritance of dioecy and made B. dioica the first organism for which XY sex determination was experimentally proven. Applying molecular tools to this system, we developed a sex-linked sequence-characterized amplified region (SCAR) marker for B. dioica and sequenced it for individuals representing the full geographic range of the species from Scotland to North Africa. For comparison, we also sequenced this marker for representatives of the dioecious B. cretica, B. multiflora and B. syriaca, and monoecious B. alba. In no case did any individual, male or female, yield more than two haplotypes. In northern Europe, we found strong linkage between our marker and sex, with all Y-sequences being identical to each other. In southern Europe, however, the linkage between our marker and sex was weak, with recombination detected within both the X- and the Y homologues. Population genetic analyses suggest that the SCAR marker experienced different evolutionary pressures in northern and southern Europe. These findings fit with phylogenetic evidence that the XY system in Bryonia is labile and suggest that the genus may be a good system in which to study the early steps of sex chromosome evolution. PMID- 19120822 TI - Egg jelly influences sperm motility in the externally fertilizing frog, Crinia georgiana. AB - Recent in vitro fertilization studies have revealed female and male x female interaction effects on the probability of fertilization. These findings suggest a mechanism of cryptic female choice via sperm-egg interactions. The egg jelly of anuran amphibians contains proteins that facilitate the chemoattraction and binding of sperm for fertilization. Here we show that egg jelly also influences the onset of motility and swimming velocity of motile sperm in the frog Crinia georgiana. Moreover, we found significant among female variation in the effects of egg jelly on sperm motility. We discuss this finding with respect to male and female effects on nonrandom fertilization observed in this species. PMID- 19120823 TI - President's Editorial--AAFS and Forensic Science: the next 60 years. PMID- 19120824 TI - Fetal age estimation using orbital measurements: 3D CT-scan study including the effects of trisomy 21. AB - This study evaluates a new method of fetal age estimation based on orbital measurement and including the potential trisomy 21 of the fetus. Six orbital and one facial CT-scan measurements were taken on 71 fetuses ranging from 14 to 41 weeks of gestational age. Forty-eight fetuses were "normal" and 23 fetuses presented the Down syndrome (trisomy 21). The anatomy/imagery correspondence was evaluated comparing our results to those obtained by direct bone measurements on the same fetuses and revealed no significant differences between the two kinds of measurements. Moreover, a multiple linear stepwise regression was realized to estimate fetal age and showed that the "trisomy 21 parameter" is not conserved in the final determination model. Therefore, we conclude that the good radio anatomical correspondence offers an interesting alternative to direct bone measurement (necessitating dissections) and allows a reliable fetal age determination, whatever is the trisomy 21 condition of the fetus. PMID- 19120825 TI - Comparative analysis of the HV1 and HV2 regions of human mitochondrial DNA by denaturing high-performance liquid chromatography. AB - Denaturing high-performance liquid chromatography (DHPLC) was evaluated as a sequencing-independent means of detecting the presence of sequence differences in pair-wise mixtures of nonconcordant amplicons of human mitochondrial DNA (mtDNA). A total of 920 pair-wise combinations of HV1 and HV2 mtDNA amplicons from 95 individuals were assayed by DHPLC for sequence concordance/nonconcordance. For the 72 combinations of amplicons from different individuals who shared identical DNA sequences, DHPLC assays consistently indicated sequence concordance between the samples. This was in 100% agreement with sequencing data. For the 849 combinations of amplicons which differed in sequence, DHPLC detected the presence of sequence nonconcordance in all but 13 assays to yield 98.5% concordance with sequencing. Thus, DHPLC can be used to detect a diversity of sequence differences (transitions, transversions, insertions, and deletions) in the mtDNA D-loop. Accordingly, DHPLC may have utility as a presumptive indicator of mtDNA sequence concordance samples, as a screen for heteroplasmy/situational mixtures, and as a means for the physical fractionation of the individual contributors to an mtDNA mixture prior to sequencing. PMID- 19120826 TI - Estimating projectile perpendicular impact velocity on metal sheet targets from the shape of the target hole. AB - The correlation between bullet hole shapes in metal and projectile impact velocity was examined. A series of shots were fired from an M-16A1 assault rifle of 5.56 mm caliber toward a 1-mm thick metal target. All shots were fired at a perpendicular angle to the metal sheets, and the velocity was measured just before the projectile hit the target. Velocities ranged between 400 and 900 m/sec. From the replica of the shooting hole, a perpendicular plane was created, showing the symmetrical properties of the hole. The best mathematical equation describing the shape of the entrance hole was the exponential function in the form: Y x = A + Be kx. The empirical equation of the hole defined using the regression method is: Y x,V = 8.268/V 0.578018 e(0.584x/V0.005). This equation describes the general shape of shooting holes created by velocities ranging from 440 to 750 m/sec. From this equation, one can estimate the bullet velocity when it hits the target. PMID- 19120827 TI - Proteolytic-based method for the identification of human growth hormone. AB - Human growth hormone (HGH) is a relatively small protein consisting of 191 amino acids and has an average mass of 22,125 amu. The forensic analysis of proteins such as HGH must meet the analytical sufficiency requirements for the laboratory and consists of a binary approach. A suspected sample is analyzed as the whole protein for retention time and mass determination using high performance liquid chromatography equipped with a photodiode array and liquid chromatography mass spectrometry. Further fragmentation of the protein using a proteolytic enzyme adds another dimension to the specificity of the analysis. Porcine trypsin digests proteins in a very predictable manner and yields peptide fragments of the original protein that can be used as a means for fingerprinting the larger biomolecule. In silico, or theoretical, digestion of HGH by trypsin yields 21 peptides ranging in size from 1 to 23 amino acids in length. The larger fragments containing higher numbers of amino acids give more specificity to identifying a protein based on a fragment produced by the digestion of trypsin. Herein, the analysis of HGH using a proteolytic approach is presented that meets the Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) recommendations for the identification of unknown substances. PMID- 19120828 TI - Fatal accident cause and conclusion. AB - The Toolmarks Laboratory prepared a report concerning a traffic accident involving the death of a road workman. The driver of the vehicle that hit the workman claimed that the wheels had failed to respond when turning left at a roundabout. A traffic investigator photographed the rack and pinion assembly of the steering system, which was subsequently removed and brought to the Toolmarks Laboratory. The rack and pinion assembly of the steering system was rebuilt, and examination showed that the system functioned properly. Specifically, the front wheels responded correctly to the steering input. Laboratory photographs of the steering system were taken in two different positions, from the same angles as the investigator's photographs at the scene of the accident. It was clear that the steering system had not been assembled properly in the garage. PMID- 19120829 TI - The application of remote sensing for detecting mass graves: an experimental animal case study from Costa Rica. AB - Detection of mass graves utilizing the hyperspectral information in airborne or satellite imagery is an untested application of remote sensing technology. We examined the in situ spectral reflectance of an experimental animal mass grave in a tropical moist forest environment and compared it to an identically constructed false grave which was refilled with soil, but contained no cattle carcasses over the course of a 16-month period. The separability of the in situ reflectance spectra was examined with a combination of feature selection and five different nonparametric pattern classifiers. We also scaled up the analysis to examine the spectral signature of the same experimental mass grave from an air-borne hyperspectral image collected 1 month following burial. Our results indicate that at both scales (in situ and airborne), the experimental grave had a spectral signature that was distinct and therefore detectable from the false grave. In addition, we observed that vegetation regeneration was severely inhibited over the mass grave containing cattle carcasses for up to a period of 16 months. This experimental study has demonstrated the real utility of airborne hyperspectral imagery for the detection of a relatively small mass grave (5 m(2)) within a specific climatic zone. Other climatic zones will require similar actualistic modeling studies, but it is clear that the applications of this technology provide the international community with both an early detection tool and a tool for ongoing monitoring. PMID- 19120836 TI - Letter to the editor--stuffed toy animals as transitional objects in an adult suicide. PMID- 19120837 TI - Commentary on: Comparison of presumptive blood test kits including Hexagon OBTI. PMID- 19120839 TI - Commentary on: "Elevated morphine concentrations determined during infant death investigations: artifacts of withdrawal of care". PMID- 19120841 TI - Commentary on: Electrocution in Sydney-a 10-year case review. PMID- 19120843 TI - Contrast-enhanced ultrasound: emerging modality for liver lesion characterization. PMID- 19120844 TI - Optimal imaging of hepatic mass lesions. PMID- 19120845 TI - Intraductal ultrasound in biliary disorders. PMID- 19120846 TI - Inflammatory cytokine gene polymorphisms and gastric cancer. PMID- 19120847 TI - Early changes of the portal tract on microcomputed tomography images in a newly developed rat model for Budd-Chiari syndrome. AB - BACKGROUND AND AIM: The effect of increased sinusoidal pressure on the portal tract in Budd-Chiari syndrome (BCS) is as yet not elucidated. Our aim was to investigate portal changes in a newly-developed rat model for BCS. METHODS: We created an outflow obstruction in Sprague-Dawley rats (n = 6) by diameter reduction of the inferior vena cava. Left and right liver lobes with portal vein contrast were scanned using microcomputed tomography, and volumes of the portal tree and liver parenchyma were computed by the ANALYZE software program. RESULTS: Portal branching density was significantly lower in BCS than the shams, and decreased over time (P < 0.01). There was a significant drop in volume of both parenchyma and the portal tree in the left but not right lobes. At 6 weeks post surgery, the perfusion index (i.e. ratio between both volumes) became equal to (left) or even higher than (right) the shams, suggesting a new equilibrium with preserved portal perfusion. Histological findings were consistent with those observed in humans. CONCLUSION: As early as day 2, a significant loss of peripheral portal branches was seen, which progressed over time. Inter-lobar differences in vascular abnormalities suggest compensatory mechanisms. Despite a decrease in both liver and portal vein volume, relative portal perfusion appeared spared. PMID- 19120848 TI - Education and imaging. Hepatobiliary and pancreatic: Caroli's disease. PMID- 19120849 TI - Education and imaging. Hepatobiliary and pancreatic: malignant gallbladder polyp. PMID- 19120851 TI - Education and imaging. Gastrointestinal: mesenteric panniculitis. PMID- 19120850 TI - Education and imaging. Gastrointestinal: mantle cell lymphoma diagnosed by balloon enteroscopy. PMID- 19120852 TI - Education and imaging. Gastrointestinal: small bowel volvulus. PMID- 19120853 TI - Adenocarcinoma of the esophagogastric junction: do we see more or less? PMID- 19120854 TI - Does the level of hepatitis B virus vaccination in health-care workers need improvement? PMID- 19120855 TI - Natural history of chronic hepatitis B virus infection: what determines prognosis after cirrhotic decompensation? PMID- 19120856 TI - Diabetes mellitus and prognosis after curative therapy for hepatocellular carcinoma: alas, still grave for those who are hyperglycemic. PMID- 19120857 TI - Endotherapy for pain in chronic pancreatitis. AB - Pain is the most distressing symptom of chronic pancreatitis. Although the pathogenesis of pain is still poorly understood, an increase in intraductal pressure may be the dominant factor. The management of pain can involve medical, endoscopic, neurolytic, and surgical therapies. Endotherapy includes pancreatic sphincterotomy, extraction of stones, placement of stent, and dilatation of strictures, sometimes preceded or followed by extracorporeal shock-wave lithotripsy. Several studies have now shown that endotherapy provides partial or complete relief of pancreatic pain in a majority of patients with an acceptable frequency of early and late complications. Endotherapy should now graduate from an experimental form of treatment to a realistic treatment option in patients with chronic or relapsing pain, particularly in the setting of calcific chronic pancreatitis. PMID- 19120858 TI - Proteomic identification of biomarkers related to Helicobacter pylori-associated gastroduodenal disease: challenges and opportunities. AB - Helicobacter pylori colonize the stomach of over half the world's population. While 80-90% H. pylori-infected individuals have clinically asymptomatic gastritis, 10-15% develop peptic ulcer, and 1-2% gastric malignancies. These variable clinical outcomes have led to an interest in prognostic indicators. The current disease paradigm suggests that host genetics and bacterial virulence both play important roles in modulating the final outcome of H. pylori infection. Elucidation of the interaction between host and bacterium is essential to clarify pathogenesis and to develop new strategies for prevention and treatment. Proteomic technology is a powerful tool for simultaneously monitoring proteins and protein variation on a large scale in biological samples. It has provided an unprecedented opportunity to survey a cell's translational landscape comprehensively, and the results may allow in-depth analyses of host and pathogen interactions. Using this high-throughput platform and taking advantage of complete sequences for both the H. pylori and the human genome in available databases, we have identified several crucial proteins that have pathogenic and prognostic potential. Among them, antibodies to AhpC and GroEs of H. pylori could be utilized for identification of patients who are at high risk of disease complications after H. pylori infection. Evolving proteomic technologies, together with appropriate clinical phenotyping and genotype information should enhance understanding of disease pathogenesis and lead to more precise prediction of variable disease outcomes. It will also facilitate development of biomarkers for diagnosis, treatment, and prevention of H. pylori infection. PMID- 19120859 TI - Changing trends in the proportion of adenocarcinoma of the esophagogastric junction in a large tertiary referral center in Japan. AB - INTRODUCTION: A dramatic increase in incidence of adenocarcinoma of the esophagogastric junction (EGJ) over the past two decades has been reported in the West. However, epidemiological data from Asian countries have not shown a similar trend. The aim of this study was to determine the incidence of adenocarcinoma of the EGJ in a cohort of consecutive patients operated on for gastric adenocarcinoma at a major cancer referral center in Japan. METHOD: We reviewed pathological reports of all patients who underwent surgery for advanced gastric adenocarcinoma between 1962 and 2005 at the National Cancer Centre Hospital in Tokyo. Adenocarcinoma of the EGJ was defined from images recorded for each patient, in accordance with the classification of Siewert and Stein. The proportion of adenocarcinoma at the EGJ among operated gastric adenocarcinoma patients was compiled at five-year intervals and serial comparison made. RESULTS: A total of 6953 patients with advanced gastric adenocarcinoma were operated on; adenocarcinoma of EGJ was found in 520 patients. The overall proportion of adenocarcinoma of the EGJ increased from 2.3% (1962-1965) to 10.0% (2001-2005). The proportion of Siewert Type II rose from 28.5% (1962-1965) to 57.3% (2001 2005), while that of Type I remained at around 1%. CONCLUSION: An increasing trend of adenocarcinoma of EGJ is observed in this study of patients operated on for gastric adenocarcinoma from 1962 to 2005 in a large tertiary referral center in Japan. PMID- 19120860 TI - The alteration of enterochromaffin cell, mast cell, and lamina propria T lymphocyte numbers in irritable bowel syndrome and its relationship with psychological factors. AB - BACKGROUND/AIMS: Psychological factors and subtle histopathological changes have been implicated in irritable bowel syndrome (IBS). The aims of the present study were to investigate whether the numbers of enterochromaffin (EC) cells, mast cells, and lamina propria T lymphocytes are altered in IBS, and evaluate the relationship of such alterations with psychological factors. METHODS: Forty-two consecutive IBS patients (M : F = 17:25, mean age 48 years) fulfilling the Rome III criteria and twelve asymptomatic healthy controls underwent rectal biopsy. Immunostaining was performed for EC cells, mast cells, and lamina propria T lymphocytes. RESULTS: The IBS group included five post-infectious (PI) IBS and 37 non-PI IBS patients. Significantly more EC cells, mast cells and lamina propria T lymphocytes were observed in PI IBS patients. Mast cells significantly increased in non-PI IBS-D (diarrhea) patients, but not in non-PI IBS-C (constipation) and non-PI IBS-M (mixed) patients. Enterochromaffin cell numbers were not significantly altered in non-PI IBS patients. Anxiety and depression scores did not differ between IBS patients with and without abnormal increase in EC cell or mast cell counts, defined as more than the mean of controls + 2 standard deviations. Enterochromaffin cell, mast cell, or lamina propria T lymphocyte numbers were poorly correlated with anxiety and depression scores in the IBS group. CONCLUSIONS: Enterochromaffin cells, mast cells, and lamina propria T lymphocytes significantly increase in PI IBS, whereas only mast cells significantly increase in non-PI IBS-D. Such histopathological changes do not seem to be directly associated with psychological factors. PMID- 19120861 TI - Utility of a 13C-methacetin breath test in evaluating hepatic injury in rats. AB - BACKGROUND AND AIM: Methacetin is thought to be a good substrate for the evaluation of different cytochrome P450 enzymatic systems of liver microsomes because of its rapid metabolism and lack of toxicity in small doses. Recent studies indicate that a methacetin breath test may be a non-invasive alternative for the evaluation of liver function since it correlates well with the severity of liver damage. It may also discriminate between different stages of liver cirrhosis and correlates with the Child-Pugh score. The application of this test in experimental liver damage in animal models has not yet been examined. This study aimed to evaluate the efficacy of the (13)C-methacetin breath test in assessing the extent of hepatic injury in models of acute liver failure, liver cirrhosis, and fatty liver in rats. METHODS: Absorption of methacetin given per os or intraperitoneally in normal rats was evaluated. The association between liver mass and (13)C-methacetin breath test results was assessed in a 70% hepatectomy rat model. Fulminant hepatic failure was induced by three consecutive intraperitoneal injections of thioacetamide, 300 mg/kg, at 24 h intervals. For induction of liver cirrhosis, rats were given intraperitoneal injections of thioacetamide, 200 mg/kg, twice a week for 12 weeks. A methionine-choline deficient diet was used for the induction of fatty liver. Rats were analyzed for (13)C-methacetin by BreathID (MBID) using molecular correlation spectrometry. BreathID continuously sampled the animal's breath for 60 min and displayed the results on the BreathID screen in real-time. RESULTS: Methacetin was absorbed well irrespective of the administration method in normal rats. Liver mass was associated with peak amplitude, complete percent dose recovery (CPDR) at 30 and 60 min and MBID peak time. A high degree of association was also demonstrated with MBID results in acute hepatitis (peak amplitude, 19.6 +/- 3.4 vs 6.3 +/- 1.63.4; CPDR30, 6.0 +/- 3.3 vs 1.2 +/- 0.5; CPDR60, 13.3 +/- 4.5 vs 3.2 +/- 1.4; and peak time, 31.0 +/- 14.9 vs 46.9 +/- 10.8 min) and liver cirrhosis (peak amplitude, 24.4 +/- 2.3 vs 15.6 +/- 6.4; CPDR30, 7.9 +/- 1.2 vs 2.7 +/- 1.0; CPDR60, 17.8 +/- 2.6 vs 8.8 +/- 2.1; and peak time, 30.2 +/- 1.5 vs 59.6 +/- 14.5 min), but not with grade of liver steatosis. CONCLUSIONS: Methacetin is well absorbed and exclusively metabolized in the liver. MBID is a sensitive test and may be a useful tool for the evaluation of functional liver mass in animal models of acute liver failure and cirrhosis. However, MBID could not distinguish between fatty liver and normal liver in rats. PMID- 19120862 TI - Education and imaging. Gastrointestinal: beef tapeworm (Taenia saginata). PMID- 19120863 TI - Education and imaging. Gastrointestinal: ascariasis. PMID- 19120864 TI - Education and imaging. Gastrointestinal: colonoscopic removal of an inverted appendix. PMID- 19120865 TI - Education and imaging. Hepatobiliary and pancreatic: cystic duct obstruction after metallic stents. PMID- 19120866 TI - Education and imaging. Hepatobiliary and pancreatic: isolated hydatid cyst of the pancreas. PMID- 19120867 TI - Road most traveled: gut-specific migration signals and leucocyte entry to the intestine. PMID- 19120868 TI - Inflammatory bowel disease, a dysregulated host-microbiota interaction: are probiotics a new therapeutic option? PMID- 19120869 TI - Individualizing therapy for pancreatic cancer. PMID- 19120870 TI - Newly acquired hepatitis C--many hurdles from diagnosis until treatment initiation. PMID- 19120871 TI - Gastroesophageal reflux disease: an Asian perspective. AB - The prevalence of gastroesophageal reflux disease (GERD) ranges from 2.5% to 7.1% in most population-based studies in Asia. There is evidence that GERD and its complications are rising, coinciding with a decline in Helicobacter pylori (H. pylori) infection. Asian GERD patients share similar risk factors and pathophysiological mechanisms with their Western counterparts. Possible causes for the lower prevalence of GERD include less obesity and hiatus hernia, a lesser degree of esophageal dysmotility, a high prevalence of virulent strains of H. pylori, and low awareness. Owing to the lack of precise translation for 'heartburn' in most Asian languages, reflux symptoms are often overlooked or misinterpreted as dyspepsia or chest pain. Furthermore, a symptom-based diagnosis with a therapeutic trial of the proton pump inhibitor (PPI) may be hampered by the high prevalence of H. pylori-related disease. The risk stratification for prompt endoscopy, use of a locally-validated, diagnostic symptom questionnaire, and response to H. pylori'test and treat' help improve the accuracy of the PPI test for diagnoses. PPI remain the gold standard treatment, and 'on-demand' PPI have been shown to be a cost-effective, long-term treatment. The clinical course of GERD is benign in most patients in Asia. The risk of progression from non erosive reflux disease to erosive esophagitis is low, and treatment response to a conventional dose of PPI is generally higher. Although H. pylori eradication may lead to more resilient GERD in a subset of patients, the benefits of H. pylori eradication outweigh the risks, especially in Asian populations with a high incidence of gastric cancer. PMID- 19120872 TI - Is there a link between food and intestinal microbes and the occurrence of Crohn's disease and ulcerative colitis? AB - The pathogenesis of Crohn's disease (CD) and ulcerative colitis (UC) is not fully understood. The interaction between intestinal environmental factors of food and intestinal microbes and the immunological system of hosts seems to be an important aspect. We have reviewed the relationship of the daily consumption of dietary animal meat and fats, dairy products, sugar, and other factors that may be linked to the occurrence of CD and UC from the literature and Japanese epidemiological data. In the present study, we reviewed the association between food and intestinal microbes and other factors contributing to the occurence of inflammatory bowel disease (IBD) from epidemiological data and case-control studies of IBD in the literature that appeared on Medline, and assessed the reports of intestinal microbes involved in the occurrence of IBD. We found several papers describing the positive association of animal meat and sweets and sugar with the occurrence of CD and UC. An analysis of Japanese epidemiological data suggested that the registered number of patients with CD or UC started to increase more than 20 years after an increased daily consumption of dietary animal meat and fats, and milk and dairy products, and after a decreased consumption of rice. Many studies implied a positive role of intestinal microbes in the occurrence of IBD. Intestinal environmental factors, such as Westernized food and intestinal microbes, seem to be involved in the increased occurrence of IBD. PMID- 19120873 TI - Probiotic administration alters the gut flora and attenuates colitis in mice administered dextran sodium sulfate. AB - BACKGROUND: Probiotics are used in the therapy of inflammatory bowel disease. This study aimed to determine whether prior administration of probiotic lactobacilli and bifidobacteria would prevent disease and change gut flora in an animal model of colitis. METHODS: Swiss albino mice received a probiotic mixture (four Lactobacillus and four Bifidobacterium species) or medium (control) for a week prior to induction of colitis by oral 4% dextran sodium sulfate (DSS) for seven days. Appropriate non-colitis controls were used. Histological damage was assessed (n = 5 per group), as was expression of mRNA for tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, transforming growth factor (TGF)-beta1 and SOCS-1 in the colonic mucosa (n = 6 per group). Secretion of TNF-alpha was measured in distal colon organ culture (n = 5-6 per group). Levels of Bacteroides, Bifidobacterium, and Lactobacillus acidophilus in feces were quantified by real time polymerase chain reaction (PCR) targeting 16S rDNA. RESULTS: Compared to untreated DSS colitis, probiotic treatment significantly reduced weight loss (P < 0.05), shifted histological damage to lesser grades of severity (P < 0.001), reduced mRNA expression of TNF-alpha and TGF-beta1 (P < 0.05), and down-regulated production of TNF-alpha from distal colon explants (P < 0.05). Colitis induced a significant reduction in the relative proportions of Bifidobacterium, Bacteroides and Lactobacillus acidophilus group bacteria in feces, and these levels were significantly increased in probiotic-treated mice compared to DSS mice (P < 0.001). CONCLUSION: Prior administration of probiotic bacteria reduced mucosal inflammation and damage in DSS-induced colitis. DSS colitis was associated with significant changes in the fecal anaerobic bacterial flora and these changes were modulated by administration of probiotic bacteria. PMID- 19120874 TI - Expression of nerve growth factor and tyrosine kinase receptor A and correlation with perineural invasion in pancreatic cancer. AB - BACKGROUND AND AIM: Perineural invasion (PNI) is one of the most common routes of invasion in pancreatic cancer and the exact mechanism is still not clear. The aim of the present study was to investigate the effect of nerve growth factor (NGF) and tyrosine kinase receptor A (TrkA) on PNI and to clarify the possible mechanism of PNI in pancreatic cancer. METHODS: Expressions of NGF/TrkA were examined in 51 human primary pancreatic cancer using immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR). The molecular findings were correlated with PNI, clinicopathological parameters and expression of Ki-67. RESULTS: Immunohistochemical analysis indicated that the presence and kind of PNI are prognostic parameters (P = 0.002,, P = 0.004). Tumors with high NGF expression exhibited more frequent presence of PNI (P = 0.033). NGF expression was significantly correlated with metastasis of lymph nodes and involvement of surgical margins (P = 0.006, 0.015). TrkA expression was significantly correlated with degree of PNI (P = 0.017). Negative correlations were found between expression of NGF/TrkA and Ki-67. As shown by RT-PCR, mRNA levels of NGF/TrkA with PNI were significantly higher than that without PNI. CONCLUSIONS: In pancreatic cancer, overexpression of NGF may contribute to PNI by prompting the hyperplasia of nerves, restraining the apoptosis of tumor cells and specifically combining NGF and TrkA. PMID- 19120875 TI - Liver histology in benign biliary stricture: fibrosis to cirrhosis . . . and reversal? AB - BACKGROUND: Secondary biliary cirrhosis is a potential complication of post cholecystectomy bile duct stricture (PCBDS). This study addresses the factors that determine the severity of pathological changes on liver biopsy and the correlation with long-term outcome following repair. METHODS: Liver biopsies obtained at surgery for repair of PCBDS in 71 patients were reviewed and pathological changes were scored from 0 to 3. Patients with fibrosis score 0-2 were categorized as the non-cirrhotic group and those with score 3 (secondary biliary cirrhosis) were categorized as the cirrhotic group. Clinical and biochemical parameters, stricture type and outcome were analyzed by univariate and multivariate analysis for correlation with degree of fibrosis. Follow-up liver biopsies (3-60 months) after stricture repair were obtained in five patients. RESULTS: There were 58 patients in the non-cirrhotic group and 13 in the cirrhotic group. On univariate analysis, portal hypertension and prolonged injury-repair duration correlated with secondary biliary cirrhosis. Patients with a fair outcome in the cirrhotic group (4/13) had derangements in liver function tests but had patent biliary enteric anastomosis on evaluation. Of the five patients in whom liver biopsies were obtained at follow up, two had regression, two were static, and one had progression. CONCLUSION: All patients with PCBDS had varying degrees of fibrosis. Prolonged injury-repair interval and portal hypertension were the important parameters correlating with secondary biliary cirrhosis. Early repair of biliary stricture is recommended to prevent liver fibrosis. A successful relief of biliary obstruction may halt and/or reverse pathological changes in the liver. PMID- 19120876 TI - Epidemiological study of hepatitis E virus infection in the general population of Okinawa, Kyushu, Japan. AB - BACKGROUND AND AIM: The aim of this study was to investigate the prevalence of hepatitis E virus (HEV) infection in the general population of Japan by determining presence of the antibody to HEV (anti-HEV). METHODS: The prevalence of HEV infection was determined by positivity of serum antibody to HEV (anti HEV). RESULTS: On retrospective analysis, a significant decrease in anti-HEV prevalence was found in Okinawa healthy residents from 1995 (15.8%) to 2005 (5.5%) (P < 0.0001). In 2005, the anti-HEV prevalence was significantly higher in Okinawa wild boar hunters (25.3%) than in the residents (male 7.7% and female 4.1%) (P < 0.0001). A significant difference was found in the history of consumption of undercooked or raw boar meat between anti-HEV positive and negative hunters (100% vs 64.3%) (P = 0.0018). CONCLUSIONS: In conclusion, the anti-HEV prevalence has decreased in the residents of this area, but HEV infection has continued at a high rate in the hunters through the custom of eating undercooked or raw boar meat. PMID- 19120877 TI - Using a surveillance system to identify and treat newly acquired hepatitis C infection. AB - BACKGROUND AND AIM: Hepatitis C treatment uptake in Australia is low. We describe the rate of acute hepatitis C treatment in the Australian state of Victoria by linking the centralized passive notifications system for hepatitis C with the Australian Trial in Acute Hepatitis C (ATAHC), a nationwide clinical trial aimed at providing people with newly acquired hepatitis C with 24 weeks' pegylated interferon monotherapy. METHODS: Mandatory notifications of clinical or laboratory evidence of hepatitis C were further investigated for evidence of newly acquired infection. Followed-up individuals were then screened for eligibility for the ATAHC study (which included documented hepatitis C antibody positivity within the previous 6 months) and offered acute hepatitis C treatment if this was the case. We examined the first 18 months of the recruitment. RESULTS: A total of 4591 hepatitis C cases were notified with 414 (9%) of these flagged as being potentially newly acquired. Through follow-up of doctors and patients, 160 of these were confirmed as newly acquired; 87 of these 160 (54%) were potentially eligible for ATAHC and were referred to ATAHC researchers. Fourteen (16%) were successfully enrolled in ATAHC. Eight individuals commenced acute hepatitis C treatment during this period. CONCLUSION: The use of hepatitis C surveillance system has been successful in identifying cases of newly acquired hepatitis C which are often difficult to identify in a clinical setting. In addition, marginalized patients who may otherwise never have been referred to a clinic are able to access hepatitis C treatment and specialist services. Despite this, only eight out of 87 eligible individuals (9%) began acute hepatitis C treatment. PMID- 19120878 TI - Is there yet any place for reagent strips in diagnosing spontaneous bacterial peritonitis in cirrhotic patients? An accuracy and cost-effectiveness study in Brazil. AB - BACKGROUND: Diagnosis of spontaneous bacterial peritonitis (SBP) is currently based on ascitic cell counting, but there is a need for a more simple and rapid diagnostic tool. The objectives of this study are to evaluate the accuracy of reagent strips in diagnosing SBP and compare their costs with total and differential cell counts. PATIENTS AND METHODS: 71 cirrhotic in- and outpatients were consecutively included (159 samples). Spontaneous bacterial peritonitis was defined as neutrophil cells >or= 250/microL. The cutoff values for each reagent strip were defined by a receiver operating characteristic (ROC) curve. Sensitivity (S), Specificity (Sp), Positive and Negative Predictive Values (PPV and NPV), Accuracy (Ac) and cost-effectiveness (US$) in comparison to cell count exam were calculated. RESULTS: Spontaneous bacterial peritonitis was diagnosed in 17 patients (23.9%), 11 of them with positive culture (64.7%). The best cutoff points found in ROC curves were 1+ for Multistix 10 SG and ca. 75 for Choiceline 10 (Multistix 10 SG S = 80%, Sp = 98.5%, PPV = 90.9%, NPV = 96.2%, Ac = 95%; Choiceline 10 S = 76.9%, Sp = 97.7%, PPV = 87%, NPV = 95.6%, Ac = 94%). In terms of cost-effectiveness by cost/accuracy, cell count was 41.5, Multistix 10 SG 0.57, and Choiceline 10, 0.19 (P < 0.001). CONCLUSION: Reagent strips are a useful tool for diagnosing SBP in cirrhotic patients, but they have some limitations. Strips are especially indicated when total and differential cell counts are not quickly available or sometimes unavailable. They are also indicated as screening test in emergency rooms to anticipate the diagnosis of SBP and allow its early treatment. It's an interesting option in developing countries. PMID- 19120879 TI - Hepatobiliary and pancreatic: biliary imaging with 3-dimensional ultrasound. PMID- 19120880 TI - Hepatobiliary and pancreatic: cirrhosis secondary to hepatic sarcoidosis. PMID- 19120881 TI - Gastrointestinal: dissecting esophageal hematoma with a Mallory-Weiss tear. PMID- 19120882 TI - Gastrointestinal: rolled-up tongue; a sign of previous caustic injury. PMID- 19120883 TI - Gastrointestinal: strangulated Meckel's diverticulum. PMID- 19120884 TI - Proceedings of the 12th Taishotoyama Symposium on Gastroenterology, April 27-28, 2007, Shimoda, Japan. PMID- 19120886 TI - Novel single nucleotide polymorphism of the VEGF gene as a risk predictor for gastroduodenal ulcers. AB - BACKGROUND AND AIM: The gastroduodenal ulcer (GDU) mostly develops on the lesser curvature side of stomach and the anterior wall of duodenal bulb, irrespective of the etiologies including Helicobacter pylori infection, non-steroidal anti inflammatory drugs, alcohol, etc. However, a clear explanation as to why ulcers are prevalent in these areas has still not been given. The current study was designed to evaluate whether the vascular endothelial growth factor (VEGF) polymorphism could predict susceptibility to GDU through deranged angiogenic activities. METHODS: A large scale case-control study based on known single nucleotide polymorphisms (SNP) of VEGF and another case control study based on the novel SNP of VEGF was performed through the SNP-IT assay using the SNP stream 25 k system. A site-directed mutagenesis and functional assay was executed to document the biological effect of a novel VEGF SNP on angiogenesis. RESULTS: Even though the case-control study between non-ulcer dyspepsia (NUD) and gastric ulcer (GU) patients was done in 10 SNP of the VEGF gene including -2488C/T, -634G/C, 7C/T, 3436G/C, 6112C/A, 6894C/T, 9374G/A, 9812C/T, 13128C/T, and 13553C/T, the analysis showed no statistically significant association between NUD and GU. Denaturing high-performance liquid chromatography analysis could identify two novel SNP of the VEGF gene, -1780T/C and IVS-99 G/C, among which -1780T/C showed a very strong association between NUD and GDU, presenting with OR=2.93 on codominant analysis (P<0.001), OR=8.62 on dominant analysis (P<0.001), and OR=3.21 on recessive analysis (P<0.001). The promoter assay using a site-directed mutagenesis and in vitro angiogenesis assay showed repressed transcription of the VEGF gene in gastric epithelial cells and defective tube formation in endothelial cells, both transfected with a plasmid containing -1780C/C mutant of VEGF gene. CONCLUSION: The novel VEGF polymorphism -1780T/C could significantly predict the predisposition to GDU after the exposure to etiologic risks, based on defective angiogenic activity. PMID- 19120887 TI - Increased pentosidine, an advanced glycation end-product, in urine and tissue reflects disease activity in inflammatory bowel diseases. AB - BACKGROUND: Under inflammatory conditions with strong oxidative stresses, advanced glycation end-products (AGE), carbonyl compounds, are produced. The concentration of pentosidine, an AGE, reportedly correlates with complications of diabetes mellitus and worsening of rheumatoid arthritis, but its role in the pathogenesis of inflammatory bowel diseases (IBD) is unclear. METHODS: Immunohistochemistry was performed with antibodies against pentosidine, and 8-OH 2-deoxyguanosine. The urinary concentration of pentosidine was also quantified by enzyme-linked immunosorbent assay method. RESULTS: Pentosidine expression was up regulated in the inflamed tissue of IBD. The expression of both pentosidine and 8 OH-2-deoxyguanosine was similar and increased in the inflamed epithelium and infiltrating cells (neutrophils and lymphocytes). The urinary concentration of pentosidine in active ulcerative colitis was significantly greater than that in inactive ulcerative colitis (0.12+/-0.15 vs 0.021+/-0.011 microg/mg of Cr, P<0.05), and was greater in active Crohn's disease than in inactive Crohn's disease (0.071+/-0.086 vs 0.039+/-0.023 microg/mg of Cr). CONCLUSIONS: The urinary pentosidine level correlated with the activity of ulcerative colitis and may be a marker for disease activity in ulcerative colitis. PMID- 19120888 TI - Changes in the expression of claudins in active ulcerative colitis. AB - BACKGROUND AND AIM: Epithelial barrier function is impaired in ulcerative colitis (UC), but the pathophysiological mechanisms leading to this barrier defect are still far from clear. Because epithelial barrier function is primarily regulated by the most apical intercellular junction, referred to as the tight junction (TJ), we investigated the expression of TJ proteins on rectal epithelial mucosa in UC. METHODS: Biopsies from the rectum of patients with active UC and of controls were studied. Tight-junction proteins were dual stained using specific antibodies (claudin-1, -2, -3, -4, and -7) as primary antibodies, and Cy-3 conjugated anti-rabbit IgG and Alexa488-conjugated anti-mouse IgG as secondary antibodies. Samples were analyzed by immunofluorescence microscopy. Immunoblotting and real-time polymerase chain reaction were performed to quantify TJ proteins and mRNA, respectively. RESULTS: At TJ, claudin-4 and -7 staining was down-regulated in active UC, whereas claudin-2 staining was up-regulated. Claudin 4 and -7 proteins were down-regulated on immunoblotting, whereas claudin-2 protein was up-regulated in active UC. Claudin-1 and -3 expression levels were unchanged in controls and active UC. In active UC claudin-2 mRNA was increased, whereas claudin-4 and -7 mRNA were decreased. CONCLUSIONS: Down-regulation of claudin-4 and claudin-7, and up-regulation of claudin-2, might lead to altered TJ structure and be related to the impaired epithelial function in active UC. PMID- 19120889 TI - Gene expression profiling in Helicobacter-induced MALT lymphoma with reference to antigen drive and protective immunization. AB - We have previously shown that long-term infection of BALB/c mice with gastric Helicobacter species results in the development of histopathological lesions that resemble those seen in patients diagnosed with gastric mucosa associated lymphoid tissue (MALT) lymphoma. This paper describes analysis of this disease at the molecular level through the use of microarray technology and immunohistochemical staining. We were able to monitor the genetic changes in the gastric mucosa characterized by distinct transcriptional signatures and correlate these with histological changes as the infection progressed from a chronic inflammatory infiltrate through to MALT lymphoma. This model system also enabled us to further dissect the role of antigen presentation and prophylactic immunization in the disease process. Antimicrobial therapy to eradicate the antigen correlated with significant reduction in pathology and major changes in the gene expression profile. Subsequent reintroduction of the antigen resulted in rapid tumor development which correlated with an increase in aggressively proliferating cells and changes in the cellular composition of the tumor. The response in vaccinated animals showed that the protected animals exhibited a strikingly different transcriptional profile compared to those of non-protected or control mice, indicating that the vaccination targeted the appropriate site leaving a long lasting signature. The genes which were most significantly up-regulated included a number of adipocyte-specific factors, such as fat-cell specific cytokines and adipocyte surface markers. This study allowed for us to highlight the significance of antigen presentation in this disease and to hypothesis mechanisms associated with protective immunity. PMID- 19120890 TI - Microcirculatory alteration in low-grade gastric mucosa-associated lymphoma by Helicobacter heilmannii infection: its relation to vascular endothelial growth factor and cyclooxygenase-2. AB - BACKGROUND: There are clinical reports that Helicobacter heilmannii, as well as Helicobacter pylori, has been clinically reported to cause gastric low-grade mucosa-associated lymphoid tissue-type (MALT) lymphoma, although its precise mechanism remains to be clarified. Thus, the present study was undertaken to elucidate the alteration of the microcirculatory structure and the relation to angiogenetic factors in mice infected with H. heilmannii for 3 and 6 months. METHODS: Immunohistochemical studies have been performed by FITC-dextran intra aortic infusion or CD31, vascular endothelial growth factor-A, cyclooxygenase 2 antibodies using our recently established model of gastric mucosa-associated lymphoid tissue-type gastric B-cell lymphoma in C57BL/6 mice. RESULTS: Increased microcirculatory network was recognized surrounding the MALT lymphoma tissues by both the FITC-dextran infusion method and CD31 immunoreactivity. Vascular endothelial growth factor-A immunoreactivity was recognized within the lymphoma tissues as well as in the marginal area, while cyclooxygenase-2 immunoreactivity was localized in the area surrounding the MALT lymphoma tissues. CONCLUSION: Increased microvascular network as well as enhanced VEGF-A immunoreactivity was shown to be related to expansion of the MALT lymphoma formed by Helicobacter heilmannii infection. PMID- 19120891 TI - Sonic hedgehog and CDX2 expression in the stomach. AB - Sonic hedgehog (Shh) is an essential regulator of patterning processes throughout development, and CDX proteins act as the master regulators for intestinal development and differentiation. Shh and CDX2 seem to be interdependently linked with cellular differentiation through different signal cascades. We have recently shown that the loss of Shh and aberrant expression of CDX2 in Helicobacter pylori (H. pylori)-associated atrophic gastritis can be modified by H. pylori eradication prior to incomplete intestinal metaplasia. On the other hand, abnormal signaling of the hedgehog pathway has been reported in gastric cancer, especially diffuse-type cancer and advanced gastric cancer, and Shh acts as a proliferation factor in both the normal mucosa and malignant lesions. CDX2 expressed in the early stage of gastric carcinogenesis is associated with the intestinal phenotypic region and thus with a better outcome. However, it remains unclear how Shh and CDX2 are involved with intestinal transformation and further carcinogenesis. PMID- 19120892 TI - Gatifloxacin-based triple therapy as a third-line regimen for Helicobacter pylori eradication. AB - BACKGROUND AND AIM: This study was designed to investigate the efficacy of gatifloxacin (GAT)-based triple therapy as a third-line treatment for Helicobacter pylori (H. pylori) eradication, according to the assessment of the susceptibility to GAT and gyrA mutation. METHODS: Fourteen patients who had eradication failure following both clarithromycin-based triple therapy and metronidazole-based triple therapy, or who were infected with H. pylori isolates that were resistant to both clarithromycin and metronidazole after failure of clarithromycin-based triple therapy, were enrolled. These patients were randomly assigned to two groups: (i) rabeprazole and amoxicillin (RA) and (ii) rabeprazole, amoxicillin, and GAT for 7 days (RAG). The minimal inhibitory concentrations were determined by the agar dilution method. The gyrA gene was examined by sequencing. RESULTS: The eradication rate was 0% in the RA group and 75% in the RAG group. The eradication rate in the RAG group was 100% in patients infected with GAT-susceptible bacteria and/or bacteria without gyrA mutations, but was only 33.3% in those infected with GAT-resistant bacteria or bacteria with gyrA mutations. CONCLUSION: Although GAT may be a promising candidate for third line therapy, its selection must be based on the results of drug susceptibility testing or gyrA analyses. PMID- 19120893 TI - Tailored eradication therapy based on fecal Helicobacter pylori clarithromycin sensitivities. AB - BACKGROUND AND AIM: Helicobacter pylori (H. pylori) eradication rates using the PPI/AC regimen (proton pump inhibitor + amoxicillin + clarithromycin) are declining. We trialed tailoring eradication regimens according to clarithromycin (CAM) susceptibility. METHODS: The subjects were 70 H. pylori positive adults. They were randomly allocated to a tailored group and a control group. In the tailored group, subjects with CAM-sensitive strains were given PPI/AC eradication therapy, and those with CAM-resistant strains were given PPI/AM (metronidazole instead of clarithromycin) therapy. The control group were all given PPI/AC therapy. CAM sensitivity was measured by collecting fecal specimens, and extracting the DNA. The 23S rRNA domain, associated with CAM susceptibility in H. pylori, was amplified using a nested polymerase chain reaction (PCR), and DNA sequencing was used to detect point mutations at A2143G and A2144G. RESULTS: Eradication rates were 94.3% in the tailored group and 71.4% in the control group. In particular, the eradication rate was 100% for CAM-resistant strains in the tailored group. CONCLUSIONS: In Japan, where CAM-resistant H. pylori strains are expected to continue to increase, tailored eradication therapy according to CAM sensitivity will be of benefit. PMID- 19120894 TI - Growth inhibitory action of cranberry on Helicobacter pylori. AB - BACKGROUND AND AIM: Cranberry is a fruit that originated in North America, and it has been used by Native Americans for bacterial infections. Recent studies have revealed it to be effective for preventing refractory urinary infections, while also suggesting that it plays a possible role in the eradication of Helicobacter pylori (H. pylori). METHODS: The H. pylori strains used in the present study were NCTC11637 and 11638. Sugar and organic acid-rich, and polyphenol-rich fractions were obtained from cranberry juice concentrate by Amberlite XAD7HP-column chromatography. The H. pylori growth inhibition was estimated by OD(660) and titration in liquid culture, and by an agar dilution plate method. The shapes of the bacteria were analyzed by scanning electron microscopy. RESULTS: Cranberry extract suppressed bacterial proliferation in a dose-dependent manner. In the comparison with other juices, polyphenol-rich fruits (cranberries, blueberries, and red grapes) showed similar growth inhibitory activity, whereas polyphenol poor fruits (oranges, pineapples, apples, and white grapes) did not show any activity. The polyphenol-rich fraction of cranberry maintained the H. pylori growth inhibitory activity. More bacteria in a coccoid form were observed after culture with cranberry. CONCLUSION: Cranberry extract inhibited H. pylori proliferation and it is suggested that polyphenols are responsible for this action. The morphological analysis suggested that cranberry induces H. pylori to develop a coccoid form, thereby inhibiting its growth bacteriostatically. Further basic studies to clarify these mechanisms in combination with in vivo studies are needed. PMID- 19120895 TI - Impact of transnasal ultrathin esophagogastroduodenoscopy (UT-EGD) in the evaluation of esophageal peristaltic function. AB - BACKGROUND: We used transnasal ultrathin esophagogastroduodenoscopy (UT-EGD) to simultaneously perform realtime esophageal manometry and observe esophageal peristalsis. METHODS: The subjects were 22 healthy volunteers and 10 patients with proton-pump inhibitor (PPI) dependent gastroesophageal reflux disease (GERD). We induced the primary peristaltic wave associated with swallowing and observed it endoscopically in the lower esophagus, at the same time measuring the intraesophageal pressure using a manometry catheter. RESULTS: The mean primary peristaltic amplitude associated with swallowing was 65.6+/-47.4 mmHg in the volunteer group, and 28.0+/-25.6 mmHg in the GERD group. Although peristalsis was observed endoscopically in the GERD group, in some cases incomplete peristalsis left a small but definite lumen and in these subjects, the primary peristaltic wave was almost flat. CONCLUSIONS: The use of an ultrathin transnasal endoscope makes possible simultaneous manometry and endoscopic observation of the esophagus. This combination should prove useful in the evaluation of esophageal peristaltic function, such as in the diagnosing of GERD. PMID- 19120896 TI - The spectra of functional gastrointestinal disorders (FGID) in a Japanese hospital outpatient department according to the ROME II Integrative Questionnaire. AB - BACKGROUND AND AIM: To investigate the spectra of functional gastrointestinal disorders (FGID) in outpatients in Japan, FGID were diagnosed and classified using ROME II. METHODS: Patients who visited Social Insurance Shiga Hospital with symptoms lasting >12 weeks over a 12-month period without organic diseases were enrolled. A total of 61 patients answered the ROME II Integrative Questionnaire. The answers were analyzed automatically by a computer program according to the Codes for Rome II Integrative Questionnaire. Age-adjusted ratios of FGID patients were investigated. RESULTS: Of 61 patients, 54 were diagnosed as having FGID. The ratio of male/female was 29/25, and the age was distributed from 26 to 92 years (mean, 57.8 years). There was no significant difference in age between the sexes. The adjusted ratio of FGID patients increased with age with significant correlation (P=0.00385). Of the 54 patients, 23 (42.6%) had overlapping FGID. A total of 85 FGID were diagnosed in 54 patients (including 15 cases of unspecified functional bowel disorder, 14 of irritable bowel syndrome [IBS], 13 of functional fecal incontinence, 12 of functional abdominal bloating, and eight of functional dyspepsia [FD]). The adjusted ratio of patients was significantly correlated to age in cases of unspecified functional bowel disorder, functional fecal incontinence, and functional abdominal bloating, but not in IBS or FD. CONCLUSIONS: The ROME II Integrative Questionnaire and automatic analyzer were useful tools in diagnosing and classifying FGID. Approximately 40% FGID patients had overlapping conditions. The adjusted ratio of patients in some FGID was significantly correlated with age. PMID- 19120898 TI - Taishotoyama Symposium Barriers to colorectal cancer screening: economics, capacity and adherence. AB - Colorectal cancer is the second leading cause of cancer death in the United States (U.S.). Fecal occult blood testing has been shown in randomized controlled trials to decrease mortality from colorectal cancer. The incidence and mortality associated with colorectal cancer has also been illustrated to be decreased with sigmoidoscopy and colonoscopy through case-control and prospective cohort studies. Current research focuses on determining which screening strategy is optimal, and how we may improve implementation. Primary screening colonoscopy may be most effective; however, this strategy would require up to 2.6 procedures per patient over their lifetime. Based on the U.S. census this equates to 7.5 million procedures annually; the current capacity in the U.S. is insufficient to provide this strategy for all eligible persons. Computed tomographic or magnetic resonance colonography (virtual colonoscopy) may be an attractive alternative, but capacity is also insufficient for implementation. Moreover, since virtual colonoscopy is a diagnostic but not therapeutic test, economic analysis has illustrated that this strategy will not be cost-effective compared to conventional colonoscopy unless it becomes much less expensive or is associated with greater adherence. Fecal DNA testing is a promising technology but current biomarkers are insufficiently sensitive to constitute a viable strategy. Newer tests such as self-propelled, self-guided endoscopes (Aer-O-Scope), 'active' endoscopes that decrease looping (Neoguide) and colon capsule endoscopy require formal evaluation through clinical trials prior to endorsement for colorectal cancer screening. Less than half of all eligible persons in the U.S. adhere to screening recommendations. Factors associated with screening adherence include the patient's level of education, income, access to health care, a family history of colon cancer, male gender and recommendation from the primary care physician. Conversely, non-adherence is associated with concern over the bowel preparation and fear of discomfort and embarrassment with the procedure. Moreover, it appears that the presence of multiple strategies may be the cause of non-adherence in many cases. Patients who are given a choice of several strategies are less likely to adhere to any strategy than patients who are recommended a single strategy. To increase adherence to colorectal cancer screening it is recommended that the patient be instructed to undergo the test to which they are most likely to adhere. Future research should focus on interventions to improve patient adherence to screening, and on developing accurate tests that will achieve high levels of adherence. PMID- 19120897 TI - Relationships between Helicobacter pylori infection status, endoscopic, histopathological findings, and cytokine production in the duodenum of Crohn's disease patients. AB - BACKGROUND: The chronic inflammatory process in patients with Crohn's disease (CD) may affect any part of the gastrointestinal (GI) tract. The pathogenesis of CD involves immunological abnormalities, including deficient or excessive expression of cytokines. We examined Helicobacter pylori infection status, endoscopic and histopathological findings, and cytokine production in the duodenum of CD patients in comparison with controls. METHODS: Thirty-eight CD patients underwent diagnostic upper GI endoscopy. Twelve age- and sex-matched health checkup examinees were used as controls. H. pylori infection status was assessed by the (13)C-urea breath test. At the time of endoscopy, two biopsy specimens each were obtained from the second portion of the duodenum, one for hematoxylin-eosin staining and immunohistochemical analysis with anti-CD68 antibody, and one for in vitro organ culture. Interleukin (IL)-6 and -8 levels were measured in organ culture supernatant by enzyme-linked immunosorbent assay. RESULTS: H. pylori infection was significantly (P<0.05) more frequent in controls (42%) than in CD patients (8%). In the duodenum, erosions or ulcers were more frequent in CD patients (53%) than in controls (8%). Mononuclear cell infiltration in the duodenum was more severe in CD patients than in controls and IL-6 production was higher, whereas IL-8 production showed no significant difference. CD68+ cells in the duodenum were more prominent in CD patients than in controls. CONCLUSIONS: H. pylori infection is unlikely in CD patients, but they show immunological abnormalities in the duodenum, possibly from innate immune responses. PMID- 19120899 TI - Identification of the expression profile of apoptotic esophageal cancer cells by adenoviral-fragile histidine triad treatment. AB - BACKGROUND: The fragile histidine triad (FHIT) functions as a tumor suppressor, and giving adenoviral-FHIT (Ad-FHIT) is thus expected to be clinically beneficial. Much attention has recently been focused on which genes are commonly regulated by Ad-FHIT, and which genes are dominant in Ad-FHIT-induced apoptotic cells. METHODS: Ad-FHIT apoptosis-induced cells (H1299 and TE4) and non-apoptosis induced cells (TE2) were used in the current experiments. The total RNA extracted from Ad-FHIT or control was labeled with Cy3-dCTP or Cy5-dCTP and hybridized with 19,192 genes on a chip. A microarray analysis for each gene was carried out with high reproducibility provided by seven independent experiments and duplicated oligos on a chip. RESULTS: We listed the upregulated genes based on the TE4:TE2 expression ratio, such as c-Src, Jak-1, and sialyltransferase, which are expected to be target pathways as well as the downregulated genes, including CASP8 and CASP10, after Ad-FHIT treatment in esophageal cancer. CONCLUSIONS: The current microarray analysis indicated that the apoptosis of esophageal cancer observed after giving Ad-FHIT was possibly induced by activation of the c-Src gene and inactivation of the CASP8 gene. PMID- 19120900 TI - What is the clinical significance of stromal angiogenesis in Barrett's esophagus? AB - BACKGROUND: Enriched blood vessels occur in the stroma of Barrett's esophagus and are related to the ease of tumor invasion in the early stage of the cancer. Hypoxia-inducible factor (HIF) is closely related with angiogenesis through expression of vascular endothelial growth factor. The aim of the present study was to determine the predictors for angiogenesis in Barrett's esophagus and evaluate their clinical significance. METHODS: Between December 2003 and May 2004, 209 patients with endoscopically and histologically proven Barrett's esophagus were enrolled. Before endoscopic examination, all participants answered structured questionnaires for gastroesophageal reflux symptoms and drug usage. HIF-1alpha and COX-2 protein expressions, cellular proliferation, and apoptosis were investigated immunohistochemically in biopsy samples taken from the esophagus of each patient. The degree of angiogenesis was determined by CD34 immunostaining analysis. Predictors for angiogenesis were evaluated with multivariate logistic regression analysis. RESULTS: Sixty (28.7%) of the 209 enrolled patients with Barrett's esophagus had a high CD34 score. Factors proven as positive predictors for a high CD34 score were presence of gastroesophageal reflux symptoms, reflux esophagitis, COX-2 protein expression, and the proliferating cell nuclear antigen (PCNA) index. Administration of proton-pump inhibitors and HIF-1alpha protein expression were not predictors. CONCLUSIONS: Reflux esophagitis and gastroesophageal reflux symptoms were positive predictors for enriched angiogenesis in the stromal portion of Barrett's esophagus, which has malignant potential because the epithelial cells express COX-2 and have accelerated cellular proliferation. PMID- 19120901 TI - Decreased levels of adiponectin in obese patients with gastroesophageal reflux evaluated by videoesophagography: possible relationship between gastroesophageal reflux and metabolic syndrome. AB - BACKGROUND: Body mass index (BMI) and obesity are reportedly associated with symptoms of gastroesophageal reflux disease (GERD). The present study was designed to investigate the effect of metabolic disorders including obesity on the levels of functional gastroesophageal reflux by videoesophagography. METHODS: Twenty-one patients with GERD-associated symptoms were examined by videoesophagography. On their initial visit, all patients completed the Japanese version of the Carlsson-Dent self-administered questionnaire (QUEST). The findings of videoesophagography were evaluated by the X-ray severity scores for gastroesophageal reflux (XRSS), which were defined for the total diagnosis of functional gastroesophageal reflux. Correlation between XRSS scores and physical or metabolic markers was evaluated. RESULTS: The mean XRSS in the QUEST-positive group (4.7+/-0.6) was significantly higher than that in the QUEST-negative group (3.3+/-0.5, P<0.05). XRSS correlated positively with BMI (P<0.05) and waist circumference (P<0.05), but negatively with high-density lipoprotein-cholesterol (P<0.05), serum adiponectin (P<0.05) and active ghrelin (P<0.05). In the multivariate analysis, serum adiponectin level, BMI and triglyceride independently affected the XRSS. CONCLUSION: Videoesophagography is a useful diagnostic modality for the evaluation of patients with GERD symptoms. Functional gastroesophageal reflux is seen in obese patients, especially with decreased levels of adiponectin. PMID- 19120902 TI - Extracellular HSP70 blocks CD40L-induced apoptosis and tubular formation in endothelial cells. AB - BACKGROUND: Recent studies have shown that CD40, a key player in angiogenesis and tubular formation, is an extracellular receptor of the heat shock protein 70 (HSP70)-peptide complex in endothelial cells. The aim of the present study was to determine the effect of extracellular HSP70 treatment on CD40L-suppressed apoptosis and CD40L-induced tubular formation in human umbilical vein endothelial cells (HUVEC). METHODS: The apoptotic index of CD40L-stimulated HUVEC with or without recombinant human HSP70 was evaluated using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay analysis. Binding of HSP70 peptide complex to CD40 on HUVEC was determined by double-labeling immunofluorescence methods. To evaluate the biological activity of CD40 engagement pretreated with rhHSP70 (0.5, 1 and 3 ng/mL), the extent of new capillary-like networking structure (tubular formation) formation in HUVEC was counted using an Olympus digital camera. Vascular invasion into MNK-28 cell clusters was assessed by counting the number of tubular structures extending from the HUVEC into growth factor-depleted Matrigel. Scores for CD34, HSP70 and CD40L expression levels in gastric cancer tissues were determined by immunostaining. RESULTS: CD40L stimulation inhibited vincristine-induced apoptosis of HUVEC in a dose-dependent manner. Extracellular HSP70 treatment significantly blocked the inhibition of apoptosis by CD40L in HUVEC exposed to vincristine. HSP70-peptide complex bound to CD40 on HUVEC. Extracellular HSP70 treatment also significantly reduced CD40L-induced tubular formation in a dose-dependent manner. HSP70 treatment also suppressed invasive tubular formation into MKN-28 cells clusters by CD40L-activated HUVEC. There was a significant relationship between CD40L expression levels and microvessel density; however, the relationship between HSP70 expression level and microvessel density in gastric cancer tissues was not significant. CONCLUSIONS: Extracellular HSP70 treatment blocks CD40L inhibition of apoptosis and CD40L induction of tubular formation in HUVEC. PMID- 19120903 TI - Increased intestinal expression of heme oxygenase-1 and its localization in patients with ulcerative colitis. AB - BACKGROUND: Heme oxygenase-1 (HO-1) is regarded as a sensitive and reliable indicator of cellular oxidative stress. Two end products of heme degradation, carbon monoxide (CO) and bilirubin, are involved in the protective role of HO-1 against oxidative injury. We have demonstrated enhanced expression of this enzyme and increased concentration of CO in experimental models of colitis, but the role of HO-1 in patients with ulcerative colitis (UC) has not been extensively investigated. The aim of the present study was to determine the intestinal levels and localization of ho-1 mRNA and HO-1 protein in patients with UC. METHODS: Eighteen patients with UC and 13 patients with colon cancer were prospectively selected from subjects who underwent colonoscopy. Biopsy specimens were obtained from the inflamed mucosa of UC patients and from the normal mucosa at least 5 cm from the margin of carcinoma. The expression of ho-1 mRNA was assayed by real time polymerase chain reaction (PCR). The colonic expression of HO-1 was determined by immunohistochemistry and western blotting using a monoclonal antibody against HO-1. RESULTS: The expression of ho-1 mRNA and HO-1 protein was significantly increased in the colonic mucosa of patients with active UC compared with normal mucosa. In the patients with active UC, mononuclear cells in the submucosa of the colon were positive for HO-1, and there was negligible staining in the epithelial cells. CONCLUSION: The present findings are evidence of the induction of HO-1 in the colon of UC patients. PMID- 19120904 TI - Endoscopic survey of low-dose-aspirin-induced gastroduodenal mucosal injuries in patients with ischemic heart disease. AB - BACKGROUND AND AIM: Low-dose aspirin is effective for the prevention of cardiovascular and cerebrovascular events, but the frequency of gastrointestinal injuries among users of low-dose aspirin in Japan is currently unknown. In the present study endoscopic examination was performed to investigate the frequency of gastroduodenal injuries associated with low-dose aspirin in patients with ischemic heart disease (IHD). METHODS: Screening upper endoscopic examinations were prospectively performed on 131 patients with IHD who were not receiving antiulcer treatment. Endoscopic findings such as ulcers and flat erosions were assessed as mucosal injuries. RESULTS: Mucosal injuries were found in 62 of 101 (61.4%) low-dose aspirin users and three of 30 (10%) nonaspirin users. There were significantly more mucosal injuries among low-dose aspirin users than among the non-users (P<0.0001). Gastroduodenal ulcers were found in 19 of 101 (18.8%) low dose aspirin users compared with one of 30 (3.3%) nonaspirin users. The frequency of mucosal injuries was not associated with the duration of aspirin treatment. CONCLUSION: Endoscopy frequently reveals low-dose aspirin-induced gastroduodenal injuries in patients with IHD. PMID- 19120905 TI - Proton pump inhibitors and recurrent bleeding in peptic ulcer disease. AB - Peptic ulcer disease (PUD) is one of the main lesions responsible for upper gastrointestinal (GI) bleeding, as well as esophageal varices and Mallory-Weiss tear. Helicobacter pylori and non-steroidal anti-inflammatory drugs (NSAIDs)/aspirin are the major responsible causes. In cases of upper GI bleeding, urgent endoscopy is performed after stabilization of vital signs. There are several modalities for controlling bleeding in PUD, such as ethanol injection or hypertonic saline with epinephrine. Recurrent bleeding occurs in 20% of patients after endoscopic therapy. The combination of endoscopic intervention and a proton pump inhibitor (PPI) is necessary to achieve hemostasis of active bleeding. It has been reported that high-dose omeprazole (80 mg bolus injection, then 8 mg/h continuous infusion for 72 h, then 40 mg/day orally for 1 week) can reduce recurrent bleeding, the need for surgery and mortality from hemorrhagic shock in patients with high-risk peptic ulcer bleeding, as compared with standard-dose omeprazole. The metabolism of PPIs is dependent upon P450 2C19 genotypes and the clinical usefulness of genotypic analysis remains to be determined. PMID- 19120906 TI - Involvement of bone marrow-derived stromal cells in gastrointestinal cancer development and metastasis. AB - BACKGROUND: The involvement of bone marrow (BM) in tumor-stroma reactions or tumor development has not been examined in a cancer allograft, which has otherwise been appropriate for assessing therapeutic modalities. We investigated the fate of BM-derived cells in colon cancer allografts and liver metastases in mice. METHODS: C57BL/6 mice were irradiated and rescued by BM transplantation from green fluorescent protein (GFP)-transgenic mice. MC38 colon cancer cells were stably transfected with the pDsRed gene in order to identify tumor cells by fluorescence. These were inoculated into the mice to generate subcutaneous allografted tumors or liver metastases. The tumors were observed under confocal microscopy and fluorescent immunohistochemistry to determine the fate of tumor versus BM-derived cells. RESULTS: GFP-positive (GFP(+)) cells were consistently identified as vimentin(+), alpha-smooth muscle actin (alphaSMA)(+), spindle shaped stromal cells in both the subcutaneous tumors and the liver metastases. GFP(+) cells of leukocyte lineage also infiltrated the tumors. Neither GFP(+) CD31(+) endothelial cells nor GFP(+) DsRed(+) cells were detected in the tumor. CONCLUSIONS: BM-derived cells frequently and consistently infiltrated the tumor allografts and metastases as interstitial cells and leukocytes. Cells derived from the fusion of BM cells and tumor cells were not observed. This model may be appropriate for the clarification of the effects of anticancer therapies and the study of BM-derived cells in tumor-host interactions. PMID- 19120907 TI - Expression of inducible nitric oxide synthase and nitric oxide-modified proteins in Helicobacter pylori-associated atrophic gastric mucosa. AB - BACKGROUND AND AIM: Induction of inducible nitric oxide synthase (iNOS) may be involved in carcinogenesis of the stomach, because nitric oxide (NO) derived from iNOS can exert DNA damage and post-transcriptional modification of target proteins. In the present study, we investigated the correlation between endoscopic findings and iNOS mRNA expression/NO-modified proteins in the gastric mucosa. METHODS: Fifty patients were prospectively selected from subjects who underwent upper gastrointestinal chromoendoscopy screening for abdominal complaints. The Helicobacter pylori (H. pylori) status of patients was determined by anti-H. pylori IgG antibody levels. We classified the mucosal area of the fundus as F0, fine small granules; F1, edematous large granules without a sulcus between granules; F2, reduced-size granules with a sulcus between granules; and F3, irregular-sized granules with extended sulcus between granules. Gastritis was graded using the visual analog scale of the Updated Sydney System. The expression of interleukin (IL)-8 and iNOS mRNA was assayed in gastric biopsy specimens by reverse transcription-polymerase chain reaction. NO-modified proteins were analyzed by Western blotting using novel monoclonal antibodies against nitrotyrosine. RESULTS: A total of 91.7% (11/12) of the F0 group was H. pylori negative, whereas 94.7% (36/38) of the F1-3 groups was H. pylori-positive. Spearman's analysis showed good correlation between the endoscopic grading and the score of chronic inflammation (r=0.764) and glandular atrophy (r=0.751). The expression of IL-8 mRNA was significantly increased in F1, F2, and F3 cases compared with the F0 group, with no significant differences among them. iNOS mRNA was significantly increased in the F3 group compared with the other groups, with increased nitration of tyrosine residues of proteins. CONCLUSION: The proposed classification by chromoendoscopy is useful for screening patients for atrophic and iNOS-expressing gastric mucosa with NO-modified proteins in H. pylori associated atrophic gastric mucosa. PMID- 19120908 TI - Normalization of pH level and gastric mucosa after eradication of H. pylori in the remnant stomach. AB - BACKGROUND: The Updated Sydney System (USS) is used to evaluate chronic gastritis and chronic atrophic gastritis (CAG) due to H. pylori infection. Here, we investigated USS scores and gastric juice pH levels in H. pylori infection positive or -eradicated patients with remnant stomach after surgery. METHODS: Gastric juice pH levels were measured using pH test-tape in 197 patients (112 H. pylori-positive and 85 H. pylori-negative after eradication) who had undergone distal gastrectomy and conventional H. pylori eradication therapy. RESULTS: In H. pylori infection-positive remnant stomach cases, gastric juice pH showed a reverse correlation with pepsinogen I/II ratio, and H. pylori infection-negative patients following eradication showed associations with the degree of atrophy and intestinal metaplasia at both the anastomosis and in the corpus. Further, pH levels in these patients were normalized time depending after the eradication in the remnant stomach. CONCLUSIONS: Eradication therapy for the remnant stomach contributes to the possible improvement of stomach conditions by controlling the pH level of gastric juice. This effect will be protective against the risk of secondary stomach carcinogenesis in the remnant stomach. PMID- 19120909 TI - Correlations among total colonoscopic findings, clinical symptoms, and laboratory markers in ulcerative colitis. AB - BACKGROUND AND AIM: Colonoscopy plays an integral role in the diagnosis, management and surveillance of ulcerative colitis (UC). In the present study we assessed the relationship between endoscopic and histological findings, clinical symptoms, and laboratory data. METHODS: We performed total colonoscopy examinations in 54 consecutive patients with UC. Seven segments (appendiceal region, cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum) were scored for endoscopic and histological activity. The patients were also evaluated using a symptom-activity index and laboratory markers: C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell (WBC) and platelet (plt) counts. RESULTS: There was a significant positive correlation between endoscopic and histological scores (r(s)=0.738), and between symptom activity score, endoscopic score (r(s)=0.444), and histological score (r(s)=0.557). Although the endoscopic and histological scores of distal colonic lesions (rectum-sigmoid, endoscopic: r(s)=0.515, histological: r(s)=0.624) correlated with clinical symptoms, there was no similar correlation for the proximal colon (appendiceal region-descending; endoscopic, r(s)=0.268, histological, r(s)=0.329). CRP, ESR, and WBC count also correlated with the sum of endoscopic and histological scores (CRP, r(s)=0.447, r(s)=0.369; ESR, r(s)=0.483, r(s)=0.589; WBC, r(s)=0.338, r(s)=0.330), whereas platelet count did not (r(s)=0.171, r(s)=0.210). In particular, CRP and ESR were well correlated with the activity of proximal colonic lesions (CRP, r(s) = 0.474, r(s)=0.480; ESR, r(s) = 0.423, r(s)=0.529) rather than with that of distal lesions (CRP, r(s)=0.236, r(s)=0.212; ESR, r(s)=0.368, r(s)=0.497). CONCLUSIONS: In this study, clinical symptoms reflected the activity of distal colonic lesions, whereas CRP and ESR reflected the activity of proximal lesions. Therefore, total colonoscopy may be indicated when CRP or ESR is elevated in UC patients in clinical remission. PMID- 19120910 TI - Evaluation of EORTC QLQ-C30 questionnaire in patients undergoing in-hospital chemotherapy for gastrointestinal cancer in Japan. AB - BACKGROUND AND AIM: There have been few studies in Japan of the utility of quality of life (QOL) questionnaires as an evaluation of chemotherapy for gastrointestinal (GI) cancer. The present study investigated whether QOL can be an indicator of the clinical benefit of chemotherapy, by analyzing the changes in the QOL scores of patients who underwent in-hospital chemotherapy for GI cancer. METHODS: From August 2004 to August 2006, 75 patients with GI cancer who were scheduled to undergo in-hospital chemotherapy were studied. The QOL score was measured with a questionnaire, the European Organization for Research Treatment of Cancer (EORTC) QLQ-C30 (version 3.0, Japanese version), before chemotherapy, and at 2 weeks and 1 month after the initiation of chemotherapy. Patients were divided into three groups according to the clinical response to chemotherapy (partial response [PR], no change [NC], and progressive disease [PD]). RESULTS: In total, 65 patients were included in the analysis. Global QOL and physical functioning scores worsened significantly at 2 weeks. Pain and constipation scores improved significantly at 1 month. Nausea and vomiting, appetite loss, and diarrhea scores worsened significantly at 2 weeks. The global QOL score improved significantly after chemotherapy in the PR group. The pain score improved significantly in the PR and NC groups after chemotherapy. QOL scores improved in the NC group to almost the same level as in the PR group, whereas QOL worsened in the PD group. CONCLUSIONS: Measurement of QOL score is a meaningful marker of the benefit of chemotherapy, other than tumor reduction. The NC group could be considered to have responded with improved QOL. PMID- 19120911 TI - Subclassification of superficial cardia cancer in relation to the endoscopic esophagogastric junction. AB - BACKGROUND: The incidence of adenocarcinoma of the gastric cardia has been reported to be increasing, but the endoscopic characteristics of the mucosal background of the tumor remain unclear. The purpose of the present study was to evaluate the relationship between the location of the adenocarcinoma according to the esophagogastric junction (EGJ) and mucosal characteristics. METHODS: Patients with superficial adenocarcinoma of the gastric cardia diagnosed pathologically were enrolled and divided into the above-EGJ group and the below-EGJ group according to tumor location. The EGJ was judged as the lower end of the esophageal longitudinal vessels. We retrospectively reviewed endoscopic findings with respect to the classification of reflux esophagitis, hiatus hernia, valvular appearance of the cardia and the pattern of atrophic gastritis. RESULTS: The incidence of reflux esophagitis in the above-EGJ group and below-EGJ group was 75.0% and 30.0%, respectively. The severity of hiatus hernia and the valvular appearance of the cardia of patients in the above-EGJ group were worse than in the patients in the below-EGJ group. The pattern of atrophic gastritis in the patients in the above-EGJ group was the closed type, whereas patients in the below-EGJ group had the open type. CONCLUSION: There were significant differences between cancers above or below the EGJ, so classification of adenocarcinoma of the gastric cardia according to location is recommended. PMID- 19120912 TI - The influence of the eradication of Helicobacter pylori on gastric ghrelin, appetite, and body mass index in patients with peptic ulcer disease. AB - BACKGROUND AND AIM: Helicobacter pylori (H. pylori) infection has been known to influence the gastric leptin and ghrelin secretion, for which the exact pathogenic role has not been documented yet. This study was designed to investigate the influence of H. pylori eradication on plasma or gastric levels of ghrelin, body mass index (BMI), and resultant levels of appetite in patients with peptic ulcer disease. METHODS: Twenty-two patients with H. pylori-associated active duodenal or gastric ulcer were treated with 7 weeks of antisecretory medication followed with 7 days of eradication regimen. The plasma and tissue ghrelin levels, tumor necrosis factor-alpha (TNF-alpha) mRNA, BMI, and appetite scale were checked before and after treatment. An additional endoscopic examination was conducted in 10 patients taking both ulcer treatment and H. pylori eradication. RESULTS: Gastric ghrelin mRNA expression was significantly increased after either ulcer healing or H. pylori eradication, whereas gastric TNF-alpha mRNA expression was decreased after ulcer treatment and H. pylori eradication. In parallel with these changes, the visual analog scales for hunger and prospective food consumption were significantly increased after ulcer healing and H. pylori eradication. An increase in BMI was not statistically related to ulcer healing and H. pylori eradication therapy. In the subgroup analysis of 10 patients performed with additional endoscopic examination, ulcer treatment was associated with increased plasma ghrelin level and tissue ghrelin expression irrelevant to H. pylori eradication. CONCLUSION: Restored tissue levels of ghrelin and improved status of appetite was achieved with gastric ulcer healing and H. pylori eradication. PMID- 19120913 TI - Confocal endomicroscopy in patients with ulcerative colitis. AB - BACKGROUND: Confocal endomicroscopy is a new technology that provides microscopic images of cellular morphology in the gastrointestinal tract in vivo. In particular, confocal microendoscopy allows histological diagnosis during the endoscopic examination. We examined patients with ulcerative colitis (UC) using confocal endoscopy and investigated the features of confocal endomicroscopic images of inflamed and non-inflamed rectal mucosa. METHODS: Seventeen patients with UC, 12 in the active phase and 5 in the non-active phase, and 14 non-UC control patients who had colon polyps or colon cancer underwent colonoscopy, during which fluorescence images of the rectal mucosa were obtained with confocal endomicroscopy. Histological findings of rectal biopsy specimens were compared with the microendoscopic images. RESULTS: Confocal microendoscopy allowed observation of cellular and subcellular structures. The crypts of normal colonic mucosa were small, round and regularly arranged, and the crypt lumens of the colonic glands were small and round. The crypts of colonic mucosa in non-active UC were small, round and slightly irregular in arrangement and the crypt lumens of the colonic glands were small and round. Inflammatory cells and capillaries were visible in the lamina propria. The crypts of colonic mucosa in active UC were large, variously shaped and irregular in arrangement. The crypt lumens of the colonic glands were large and the crypt structure could not be recognized in some areas. Numerous inflammatory cells and capillaries were visible in the lamina propria. CONCLUSIONS: Images taken with the confocal microendoscope provided information that was equivalent to conventional histology, and improve our understanding of the differences in rectal tissues (crypts, capillaries and inflammatory cells) between normal controls and active and non-active UC patients during ongoing endoscopy. PMID- 19120914 TI - Bacteriophages and their application in food safety. AB - In recent years it has become widely recognized that bacteriophages have several potential applications in the food industry. They have been proposed as alternatives to antibiotics in animal health, as biopreservatives in food and as tools for detecting pathogenic bacteria throughout the food chain. Bacteriophages are viruses that only infect and lyse bacterial cells. Consequently, they display two unique features relevant in and suitable for food safety. Namely, their safe use as they are harmless to mammalian cells and their high host specificity that allows proper starter performance in fermented products and keeps the natural microbiota undisturbed. However, the recent approval of bacteriophages as food additives has opened the discussion about 'edible viruses'. In this review, we examine the promising uses of phages for the control of foodborne pathogens and the drawbacks on which more research is needed to further exploit these biological entities. PMID- 19120915 TI - Chitinolytic activity of endophytic Streptomyces and potential for biocontrol. AB - AIMS: Biological sources for the control of plant pathogenic fungi remain an important objective for sustainable agricultural practices. Actinomycetes are used extensively in the pharmaceutical industry and agriculture owing to their great diversity in enzyme production. In the present study, therefore, we evaluated chitinase production by endophytic actinomycetes and the potential of this for control of phytopathogenic fungi. METHODS AND RESULTS: Endophytic Streptomyces were grown on minimum medium supplemented with chitin, and chitinase production was quantified. The strains were screened for any activity towards phytopathogenic fungi and oomycetes by a dual-culture in vitro assay. The correlation between chitinase production and pathogen inhibition was calculated and further confirmed on Colletotrichum sublineolum cell walls by scanning electron microscopy. CONCLUSIONS: This paper reports a genetic correlation between chitinase production and the biocontrol potential of endophytic actinomycetes in an antagonistic interaction with different phytopathogens, suggesting that this control could occur inside the host plant. SIGNIFICANCE AND IMPACT OF THE STUDY: A genetic correlation between chitinase production and pathogen inhibition was demonstrated. Our results provide an enhanced understanding of endophytic Streptomyces and its potential as a biocontrol agent. The implications and applications of these data for biocontrol are discussed. PMID- 19120916 TI - Detection and monitoring of anaerobic rumen fungi using an ARISA method. AB - AIM: To develop an automated ribosomal intergenic spacer region analysis (ARISA) method for the detection of anaerobic rumen fungi and also to demonstrate utility of the technique to monitor colonization and persistence of fungi, and diet induced changes in community structure. METHODS AND RESULTS: The method could discriminate between three genera of anaerobic rumen fungal isolates, representing Orpinomyces, Piromyces and Neocallimastix species. Changes in anaerobic fungal composition were observed between animals fed a high-fibre diet compared with a grain-based diet. ARISA analysis of rumen samples from animals on grain showed a decrease in fungal diversity with a dominance of Orpinomyces and Piromyces spp. Clustering analysis of ARISA profile patterns grouped animals based on diet. A single strain of Orpinomyces was dosed into a cow and was detectable within the rumen fungal population for several weeks afterwards. CONCLUSIONS: The ARISA technique was capable of discriminating between pure cultures at the genus level. Diet composition has a significant influence on the diversity of anaerobic fungi in the rumen and the method can be used to monitor introduced strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Through the use of ARISA analysis, a better understanding of the effect of diets on rumen anaerobic fungi populations is provided. PMID- 19120917 TI - Maple sap as a rich medium to grow probiotic lactobacilli and to produce lactic acid. AB - AIMS: To demonstrate the feasibility of growing lactobacilli and producing lactic acid using maple sap as a sugar source and to show the importance of oligosaccharides in the processes. METHODS AND RESULTS: Two maple sap samples (Cetta and Pinnacle) and purified sucrose were used as carbon sources in the preparation of three culture media. Compared with the sucrose-based medium, both maple sap-based media produced increased viable counts in two strains out of five by a factor of four to seven. Maple sap-based media also enhanced lactic acid production in three strains. Cetta sap was found to be more efficient than Pinnacle sap in stimulating lactic acid production and, was also found to be richer in various oligosaccharides. The amendment of the Pinnacle-based medium with trisaccharides significantly stimulated Lactobacillus acidophilus AC-10 to grow and produce lactic acid. CONCLUSIONS: Maple sap, particularly if rich in oligosaccharides, represents a good carbon source for the growth of lactobacilli and the production of lactic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a proof-of-concept, using maple sap as a substrate for lactic acid production and for the development of a nondairy probiotic drink. PMID- 19120918 TI - Efficiency of KrCl excilamp (222 nm) for inactivation of bacteria in suspension. AB - AIMS: To examine the killing efficiency of UV KrCl excilamp against gram-positive and gram-negative bacteria. METHODS AND RESULTS: Vegetative cells of Bacillus cereus, Bacillus subtilis, Escherichia coli O157:H7, Staphylococcus aureus and Streptococcus pyogenes at initial populations from 10(2) to 10(7) colony-forming units (CFU) ml(-1) were treated by KrCl excilamp in sterile Ringer's solution with and without H(2)O(2). The number of viable cells was determined using spread plating techniques and nutrient agar method with subsequent incubation at 28 degrees C or 37 degrees C for 24 h. At estimated populations of 10(2)-10(5) CFU ml(-1) E. coli O157:H7 and Staph. aureus were the most sensitive and showed 100% disinfection within 15 s (29.2 mJ cm(-2)). Bacillus subtilis was more sensitive to UV treatment than B. cereus. The UV/H(2)O(2) inactivation rate coefficients within this population range were two times higher than those observed for UV treatment alone. No effect of H(2)O(2) was observed at 10(7) CFU ml(-1) for Bacillus sp. and Strep. pyogenes. CONCLUSIONS: The narrow-band UV radiation at 222 nm was effective in the rapid disinfection of bacteria in aqueous suspensions. SIGNIFICANCE AND IMPACT OF THE STUDY: KrCl excilamps represent UV sources which can be applied for disinfection of drinking water in advanced oxidation processes. PMID- 19120919 TI - Comparison of environmental sampling methods for detecting Salmonella in commercial laying flocks in the UK. AB - AIMS: To investigate the performance of the Salmonella National Control Programme (NCP) sampling/testing methods in laying flocks of domestic fowl. METHODS AND RESULTS: Eighty-five visits were made to 69 flocks representative of the main production systems (cage, barn and free-range) infected with Salmonella. In each visit, three methodologies were compared: (i) the European Union (EU) baseline survey method (five faeces and two dust samples); (ii) an in-house (Veterinary Laboratories Agency, VLA) 'wet' method that involved collecting 10 dust and 10 faeces samples into jars with buffered peptone water; and (iii) a method involving two samples of pooled faeces and one of dust (cultured as one sample of each type), which has been adopted for the NCP for laying flocks across the EU. CONCLUSIONS: The 'wet' method was the most sensitive, and the NCP the least, although individual NCP samples were the most sensitive ones. SIGNIFICANCE AND IMPACT OF THE STUDY: The apparent lower sensitivity of the NCP method may be compensated by repeated sampling of flocks (twice during rear and several times during lay). Sampling using VLA methodology should be advocated for farms aiming to disclose low-level Salmonella before restrictions on the sale of eggs from Salmonella Enteritidis or Salmonella Typhimurium-infected flocks are in place. PMID- 19120920 TI - Tetracycline-resistance genes in gram-negative isolates from estuarine waters. AB - AIMS: To investigate the diversity and dissemination of tetracycline resistance genes in isolates from estuarine waters. METHODS AND RESULTS: Forty-two out of 164 multi-resistant isolates previously obtained were resistant or less susceptible to tetracycline, as evaluated by the disc diffusion method. Minimal inhibitory concentration for resistant bacteria ranged from 16 to 256 mg l(-1). Screening of tet genes by polymerase chain reaction showed that 88% of the isolates carried at least one of the genes tested, namely tet(A) (present in 13 isolates), tet(B) (present in 13 isolates), tet(C) (present in 3 isolates), tet(D) (present in 1 isolate), tet(E) (present in 6 isolates) and tet(M) (present in 1 isolate). One isolate carried tet(A) and tet(M). To our knowledge, this study presents the first description of a tet(D) gene in Morganella morganii. Hybridization revealed that tet genes were plasmid-located in 31% of the isolates. Those isolates were included as donors in conjugation experiments and 38% transferred tetracycline resistance. CONCLUSIONS: A considerable diversity of tet genes was detected in the estuary. Frequently, these genes were associated with plasmids and could be transferred to Escherichia coli. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented provide further evidence of the role played by estuarine reservoirs in antibiotic resistance maintenance and dissemination. PMID- 19120921 TI - A new approach for improving cordycepin productivity in surface liquid culture of Cordyceps militaris using high-energy ion beam irradiation. AB - AIMS: To obtain a higher cordycepin production using Cordyceps militaris mutant obtained by a new mutagenesis technique called 'ion beam'. METHODS AND RESULTS: Successful irradiation of C. militaris NBRC 9787 by a proton beam with high energy was performed, and 30 classes of 8-azaadenine- and 28 classes of 8 azaaguanine-resistant mutants were obtained on mutant screening, of which seven classes were selected as promising preliminary mutants having an antibacterial ability as an index of cordycepin production. In a surface liquid culture technique, some of the 8-azaadenine-resistant mutants gave a better performance for the cordycepin productivity; in contrast, among the 8-azaaguanine-resistant mutants, it was shown that mutant no. G81-3 was much better than the control in the metabolic rate of glucose and the cordycepin productivity. In primary optimization using the enriched medium, the cordycepin production was 3.1 and 1.8 g l(-1) on 21-day culture for mutant no. G81-3 and the control, respectively. The cordycepin production obtained by the mutant was 72% more than the control. CONCLUSIONS: The mutant obtained by proton beam irradiation had higher productivity of cordycepin than that of the control. SIGNIFICANCE AND IMPACT OF THE STUDY: The mutant obtained by irradiation had a superior production performance of cordycepin, and therefore, it could be used in the realm of applied industrial biotechnology for the large-scale production of cordycepin. PMID- 19120922 TI - False-negative beta-D-glucuronidase reactions in membrane lactose glucuronide agar medium used for the simultaneous detection of coliforms and Escherichia coli from water. AB - AIMS: Testing for beta-D-glucuronidase activity has become the basis of many methods for the detection of Escherichia coli in both food and water. Used in combination with tests for the presence of beta-D-glucuronidase, these tests offer a simple method for simultaneously detecting coliforms and E. coli. The purpose of this study was to determine the effectiveness of several different procedures in detecting beta-D-glucuronidase activity and hence in detecting E. coli. METHODS AND RESULTS: The ability of membrane lactose glucuronide agar (MLGA), Colilert-18, MI agar, Colitag and Chromocult agar to detect beta-D glucuronidase activity was tested with over 1000 isolates of E. coli recovered from naturally contaminated water samples. Four of the media gave very similar results but MLGA failed to detect glucuronidase activity in 15.6% of the cultures tested. CONCLUSIONS: MLGA had very poor sensitivity for the detection of beta-D glucuronidase activity in strains of E. coli isolated from naturally contaminated water. This is probably because of the fact that beta-D-glucuronidase activity is pH-sensitive and that acid is formed by E. coli during fermentation of lactose in MLGA. SIGNIFICANCE AND IMPACT OF THE STUDY: The detection of E. coli in drinking water is the primary test used to establish faecal contamination. The poor sensitivity of MLGA in detecting beta-D-glucuronidase activity suggests that this medium and others containing high concentrations of fermentable carbohydrate should not be used for the detection of E. coli. PMID- 19120923 TI - Microbiological challenge of four protective devices for the reconstitution of cytotoxic agents. AB - AIMS: To evaluate the susceptibility to microbial contamination that occurs during simulated handling of protective devices for the preparation of cytotoxic drug solutions. METHODS AND RESULTS: Four devices, i.e. Chemoprotect spike, Clave connector, PhaSeal and Securmix were challenged with low and high inocula of micro-organisms. The cells, transferred to the connected vials during repeated manipulations of the devices were counted by means of solid-phase cytometry. Of the four devices, PhaSeal afforded the lowest transfer of micro-organisms. Secondly, the efficiency of procedures for the disinfection of an artificially contaminated rubber stopper was compared prior to connection of the vial to the PhaSeal device. Spraying or swabbing alone was inadequate, as opposed to a combination of spraying [0.5% or 2.0% (w/v) chlorhexidine in isopropanol] and swabbing [70% (v/v) isopropanol]. CONCLUSIONS: Although Phaseal afforded the lowest transfer of micro-organisms, adequate disinfection of the vial prior to connection remains required. SIGNIFICANCE AND IMPACT OF THE STUDY: Unlike aspects of operator protection, which are well documented, the microbiological safety of protective devices for the preparation of cytotoxic drugs has not been addressed in the literature. This study estimates the susceptibility to microbial contamination during handling of four commonly used devices. PMID- 19120924 TI - Culture of Staphylococcus xylosus in fish processing by-product-based media for lipase production. AB - AIMS: The objective of this study was to demonstrate that fish-processing by products could be used as sole raw material to sustain the growth of Staphylococcus xylosus for lipase production. METHODS AND RESULTS: Bacterial growth was tested on supernatants generated by boiling (100 degrees C for 20 min) of tuna, sardine, cuttlefish and shrimp by-products from fish processing industries. Among all samples tested, only supernatants generated from shrimp and cuttlefish by-products sustained the growth of S. xylosus. Shrimp-based medium gave the highest growth (A(600) = 22) after 22 h of culture and exhibited the maximum lipase activity (28 U ml(-1)). This effect may be explained by better availability of nutrients, especially, in shrimp by-products. Standard medium (SM) amendments to sardine and tuna by-product-based media stimulated the growth of S. xylosus and the highest A(600) values were obtained with 75% SM. Lipase activity, however, remained below 4 U ml(-1) for both sardine and tuna by-product based media. CONCLUSIONS: Fish by-products could be used for the production of highly valuable enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of fish by products in producing S. xylosus-growth media can reduce environmental problems associated with waste disposal and, simultaneously, lower the cost of biomass and enzyme production. PMID- 19120925 TI - Influence of baking enzymes on antimicrobial activity of five bacteriocin-like inhibitory substances produced by lactic acid bacteria isolated from Lithuanian sourdoughs. AB - AIM: To evaluate the effect of four different baking enzymes on the inhibitory activity of five bacteriocin-like inhibitory substances (BLIS) produced by lactic acid bacteria (LAB) isolated from Lithuanian sourdoughs. METHODS AND RESULTS: The overlay assay and the Bioscreen methods revealed that the five BLIS exhibited an inhibitory effect against spore germination and vegetative outgrowth of Bacillus subtilis, the predominant species causing ropiness in bread. The possibility that the observed antibacterial activity of BLIS might be lost after treatment with enzymes used for baking purposes was also examined. CONCLUSIONS: The enzymes tested; hemicellulase, lipase, amyloglucosidase and amylase had little or no effect on the majority of the antimicrobial activities associated with the five BLIS studied. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests a potential application in the sourdough baking industry for these antimicrobial producing LAB strains in the control of B. subtilis spore germination and vegetative outgrowth. PMID- 19120926 TI - Effects of acacia (Acacia auriculaeformis A. Cunn)-associated fungi on mustard (Brassica juncea (L.) Coss. var. foliosa Bailey) growth in Cd- and Ni contaminated soils. AB - AIMS: To evaluate the effect of Acacia auriculaeformis-associated fungi on the growth of mustard [Brassica juncea (L.) Coss. var. foliosa Bailey] in Cd- and Ni contaminated soils and design novel plant-fungi associations for bioremediation purpose. METHODS AND RESULTS: Endophytic Trichoderma H8 and rhizosphere Aspergillus G16 were applied for rhizoremediation of Cd-, Ni-, and Cd-Ni combination-contaminated soils through association with B. juncea (L.) Coss. var. foliosa. Compared with the noninoculated control plants, inoculation with Trichoderma H8 produced 109%, 41% and 167% more fresh weight (FW) plant yields in the Cd-, Ni-, and Cd-Ni-contaminated soils, respectively (P < 0.05). Similarly, plants inoculated with Aspergillus G16 produced 109%, 47% and 44% more FW plant yields in these contaminated soils, respectively. Plants co-inoculated with these two strains produced 118%, 100% and 178% more FW plant yields, respectively. The inoculations also increased the translocation factors and metal bioconcentration factors. CONCLUSIONS: The efficiency of phytoextraction for B. juncea (L.) Coss. var. foliosa was enhanced after inoculating with Acacia-associated fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of plant-fungi association may be a promising strategy to remediate metal-contaminated soils. PMID- 19120927 TI - Comparison of TEMPO EC and TBX medium for the enumeration of Escherichia coli in cheese. AB - AIMS: TEMPO EC (Escherichia coli) is based on glucuronidase activity and is a test for use with the TEMPO system for the automated 24 h enumeration of E. coli in food products. In this study, TEMPO EC was compared with TBX medium, the current standard plate method for the enumeration of E. coli in cheese. METHODS AND RESULTS: For comparative purposes, both naturally (92) and artificially contaminated (31) cheese samples were studied. Pearson correlation coefficients were determined as 0.954 and 0.978 between the two methods for naturally and artificially contaminated samples, respectively. Regression analysis yielded the following equations: log(10) TEMPO EC = 0.340 + 0.889 log(10) TBX medium and log(10) TEMPO EC = 0.174 + 0.899 log(10) TBX medium for naturally and artificially contaminated samples, respectively. In general, absolute differences did not exceed one log between results obtained by the two methods. CONCLUSIONS: Statistical analysis of the results showed good agreement between the two enumeration methods. SIGNIFICANCE AND IMPACT OF THE STUDY: TEMPO EC is a practical and reliable alternative to the current standard plate method for the enumeration of E. coli in foods. PMID- 19120928 TI - Comparison of solid-phase cytometry and the plate count method for the evaluation of the survival of bacteria in pharmaceutical oils. AB - AIM: To compare the survival of four bacterial strains (Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa) in pharmaceutical oils, including jojoba oil/tea tree oil, carbol oil, jojoba oil and sesame oil. METHODS AND RESULTS: Oils were spiked with the test bacteria in a concentration of 10(4) CFU ml(-1). Bacteria were extracted from oils with phosphate-buffered saline containing 0.5% Tween 20. Aliquots of the pooled water layers were analysed by solid-phase cytometry and plate counting. Plate counts dropped to zero for all test strains exposed for 24 h to three of the four oils. In contrast, significant numbers of viable cells were still detected by SPC, except in the jojoba oil/tea tree oil mixture and partly in sesame oil. CONCLUSIONS: Exposure of bacteria for 24 h to the two oils containing an antimicrobial led to a loss of their culturability but not necessarily of their viability. The antibacterial activity of the jojoba oil/tea tree oil mixture supersedes that of carbol oil. SIGNIFICANCE AND IMPACT OF THE STUDY: These in vitro data suggest that the jojoba oil/tea tree oil mixture more than carbol oil inhibits bacterial proliferation when used for intermittent self-catherization. PMID- 19120929 TI - Antitumour and antimicrobial activities of endophytic streptomycetes from pharmaceutical plants in rainforest. AB - AIMS: The aim of this study was to screen antitumour and antimicrobial activities of endophytic actinomycetes isolated from pharmaceutical plants in rainforest in Yunnan province, China. METHODS AND RESULTS: Antitumour activity was studied by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and antimicrobial activity was determined by agar well diffusion method. The high bioactive endophytic isolates were identified and further investigated for the presence of polyketide synthases (PKS-I, PKS-II) and nonribosomal peptide synthetases (NRPS) sequences by specific amplification. The molecular identification confirmed that the 41 isolates showed significant activities were members of the genus Streptomyces. Among them, 31.7% of endophytic streptomycete cultures were cytotoxic against A549 cells, 29.3% against HL-60 cells, 85.4% against BEL-7404 cells, 90.2% against P388D1 cells, 65.9% were active against Escherichia coli, 24.4% against Staphylococcus aureus, 31.7% against Staphylococcus epidermidis, 12.2% against Candida albicans and no strain displayed antagonistic activity against Klebsiella pneumoniae. High frequencies of positive PCR amplification were obtained for PKS-I (34.1%), PKS-II (63.4%) and NRPS (61.0%) biosynthetic systems. CONCLUSIONS: Many endophytic streptomycetes isolated from pharmaceutical plants in rainforest possess remarkable and diverse antitumour and antimicrobial bioactivities. SIGNIFICANCE AND IMPACT OF THE STUDY: These endophytic streptomycetes are precious resources obtained from rainforests, and they could be a promising source for bioactive agents. PMID- 19120930 TI - Anti-listerial activity of bacteriocin-producing Lactobacillus curvatus CWBI-B28 and Lactobacillus sakei CWBI-B1365 on raw beef and poultry meat. AB - AIM: The study aimed to evaluate the effect of the bacteriocins produced by Lactobacillus sakei CWBI-B1365 and Lactobacillus curvatus CWBI-B28 on the growth and survival of Listeria monocytogenes in raw beef and poultry meat. METHODS AND RESULTS: The sakacin P and sakacin G structural genes were identified in Lact. curvatus CWBI-B28 and Lact. sakei CWBI-B1365 using PCR amplification, respectively. The effect of the two bacteriocinogenic strains either alone or together, and that of the nonbacteriocin-producing strain Lact. sakei LMG17302, on the growth of L. monocytogenes was evaluated in beef and poultry meat. In raw beef, the pathogenic bacteria were inhibited by the bacteriocinogenic strains. The bacteriocinogenic strains had no activity in raw chicken meat when inoculated separately, while they showed a clear anti-Listeria effect when applied together. CONCLUSION: Sakacin G producing Lact. sakei and sakacin P producing Lact. curvatus may be applied in raw beef to inhibit L. monocytogenes. In poultry meat, the inhibition of L. monocytogenes could only be achieved by a combined application of these bacteriocin-producing strains. SIGNIFICANCE AND IMPACT OF THE STUDY: In some meat products, the combined application of different class IIa bacteriocin producing lactic acid bacterium can enhance the anti-listerial activity. PMID- 19120931 TI - Polyhydroxyalkanoates production from carbohydrates by a genetic recombinant Aeromonas sp. AB - AIMS: To develop an Aeromonas strain able to utilize inexpensive carbon sources such as starch for the synthesis of polyhydroxyalkanoates (PHA). METHODS AND RESULTS: A recombinant Aeromonas sp. (strain KC007-1) was constructed by introducing the PHB synthesis genes (phaCAB) into the bacterium. Strain KC001-R1 can not only use carbohydrate (including starch) for growth but also accumulate significant amounts of polyhydroxybutyrate (PHB) in the cells. CONCLUSIONS: One of the present focuses on PHA production has been on lowering the production costs. Starch is an example of an inexpensive carbohydrate for use in industrial production of PHA. We have demonstrated that by introducing the phaCAB operon into Aeromonas sp. allowed the bacterium able to accumulated PHB using this substrate. SIGNIFICANCE AND IMPACT OF THE STUDY: Aeromonas spp. are able to synthesize PHA using fatty acids as carbon source. Although good robust growth results with use of starch as sole carbon source for Aeromonas, PHA synthesis does not occur. Strain KC007-R1 showed the ability to accumulate PHA in relative high amount with both carbohydrates and fatty acids as carbon source, and can be cultivated to a significant amount of cell mass and hence is a potential strain for further development for industrial applications. PMID- 19120932 TI - Determination of lactic acid bacteria producing biogenic amines in wine by quantitative PCR methods. AB - AIMS: To develop rapid methods allowing enumeration of lactic acid bacteria producing biogenic amines in wines and to analyse wine samples by the methods. METHODS AND RESULTS: Methods based on quantitative PCR targeting bacterial genes involved in histamine, tyramine and putrescine production were developed and applied to detect and quantify the bacteria producing these biogenic amines in wine. Analysis of 102 samples revealed low populations of the targeted bacteria in grape must samples, an increased bacteria biomass in wine samples after alcoholic fermentation, reaching the highest population levels (above 10(6) cells ml(-1)) during spontaneous malolactic fermentation. A minimum of 10(3) ml(-1) producing cells was required for production of more than 1 mg l(-1) of biogenic amines. Accumulation of putrescine in wine was correlated with the presence of bacteria carrying an ornithine decarboxylation pathway. Trials of winemaking showed that the use of selected bacteria for inducing malolactic fermentation was efficient to limit the proliferation of undesirable bacteria and the production of biogenic amines. CONCLUSION: Methods using quantitative PCR are efficient to enumerate biogenic amines-producing cells in wine. SIGNIFICANCE AND IMPACT OF THE STUDY: The methods can help to better control and to improve winemaking conditions in order to avoid biogenic amine production. PMID- 19120933 TI - Production and characterization of a low-molecular-weight bacteriocin from Bacillus licheniformis MKU3. AB - AIMS: Enhancing production and characterization of a low-molecular-weight bacteriocin from Bacillus licheniformis MKU3. METHODS AND RESULTS: The culture supernatant of B. licheniformis MKU3 exhibited bacteriocin-like activity against gram-positive and -negative bacteria and different fungi and yeast. SDS-PAGE analysis of the extracellular proteins of B. licheniformis MKU3 revealed a bacteriocin-like protein with a molecular mass of 1.5 kDa. This bacteriocin activity was found to be stable under a pH range of 3.0-10.0 and at temperatures up to 100 degrees C for 60 min, but inactivated by proteinase K, trypsin or pronase E. An experimental fractional factorial design for optimization of production medium resulted in a maximum activity of bacteriocin (11,000 AU ml( 1)) by B. licheniformis MKU3. CONCLUSIONS: A low-molecular-weight bacteriocin like protein from B. licheniformis MKU3 exhibited a wide spectrum of antimicrobial activity against several gram-positive bacteria, several fungi and yeast. A 3.6-fold increase in the production of bacteriocin was achieved using the culture medium optimized through a fractional factorial design. SIGNIFICANCE AND IMPACT OF THE STUDY: A bacteriocin with wide spectrum of activity against gram-positive bacterial pathogens, filamentous fungi and yeast suggested its potential clinical use. Statistical method facilitated optimization of cultural medium for the improved production of bacteriocin. PMID- 19120934 TI - Wide variation in effectiveness of laboratory disinfectants against bacteriophages. AB - AIMS: The purpose of this study was to identify an effective disinfectant for the inactivation of the bacteriophages (phages) being used in our laboratory, as published studies on phage inactivation are far from unanimous in their conclusions. METHODS AND RESULTS: The phages studied were three closely related strains of Myoviridae and three strains of Siphoviridae. Three disinfectants which are used commonly in microbiology laboratories were evaluated: Virkon (1%), ethanol (75%) and sodium hypochlorite (2500 ppm available chlorine). The most effective of these was Virkon, which inactivated all six phages rapidly. Ethanol was effective against the Myoviridae but had little effect on the Siphoviridae. Sodium hypochlorite was the least effective of the disinfectants evaluated. CONCLUSIONS: The findings of this study demonstrate a wide diversity in the effectiveness of disinfectants tested for inactivation of phages. SIGNIFICANCE AND IMPACT OF THE STUDY: Of the disinfectants tested Virkon is the most suitable choice for those unable to carry out disinfection validation studies, or where a broad spectrum disinfectant against phages is required. All of the phages in this study showed resilience to inactivation by sodium hypochlorite, and therefore this disinfectant is an unwise choice for use against phage without first assessing its effectiveness. PMID- 19120937 TI - The Clip score: is it time to be clipped away? PMID- 19120938 TI - Propensity for non-alcoholic fatty liver disease: more evidence for ethnic susceptibility. PMID- 19120939 TI - Clinical scoring systems for predicting outcome after surgery for colorectal liver metastases: towards a better multidisciplinary approach. PMID- 19120940 TI - Management of advanced hepatocellular carcinoma in the era of targeted therapy. AB - Systemic chemotherapy has had a disappointing track record in the management of advanced hepatocellular carcinoma (HCC). Single-agent doxorubicin produces a response rate of 10-15%, but without any survival benefit, and combination chemotherapy has also yielded unimpressive results. With recent advances in the knowledge of hepato-carcinogenesis, there has been encouraging development in the systemic therapy of advanced HCC patients, and particularly in the targeted therapy of advanced HCC. Among the newly identified targets, exciting results have been shown in targeting the anti-angiogenic pathway and the Raf/mitogen activated protein kinase pathways. Bevacizumab, both as a single agent and in combination with other agents, has shown initial encouraging activity in treating advanced HCC. More recently, single-agent sorafenib, a putative multitargeted kinase inhibitor, has shown to prolong the overall survival of patients with advanced HCC in the pivotal phase III Sorafenib HCC Assessment Randomized Protocol (SHARP) and Oriental study. Currently, sorafenib is the only approved targeted therapy for patients with advanced HCC. In addition, however, promising early results have been reported for other molecular-targeted drugs including erlotinib and sunitinib. Future progress seems likely to depend on using controlled clinical trials to optimize synergistic combination treatments. PMID- 19120941 TI - Surveillance programmes for hepatocellular carcinoma: evident-based medicine is not evidence-based medicine. PMID- 19120943 TI - Resolving the 50-year debate around using and misusing Likert scales. PMID- 19120944 TI - Thinking 'no' but saying 'yes' to student presence in general practice consultations: politeness theory insights. PMID- 19120945 TI - More students, less capacity? An assessment of the competing demands on academic medical staff. AB - CONTEXT: Medical student numbers in England have increased by more than 60% over the last decade. The capacity of universities and the National Health Service (NHS) to deliver education and training to these expanded cohorts is not clear. METHODS: We carried out an interview study in three English medical schools, involving 60 academic, administrative, clinical and managerial staff, sampled across disciplines. RESULTS: Given the expansion in medical student numbers, capacity in medical schools must increase. This requirement has become even more immediate in light of the shift towards more resource-intensive curricula. However, the aims of the Research Assessment Exercise and NHS policies are at odds with attempts to build teaching capacity. Although monies have been made available to recruit new staff and to backfill clinical time spent teaching, the success of these strategies is questioned by interviewees. Other initiatives, such as the new consultant contract and educational quality assurance processes, have the potential to promote the importance of teaching but are presently perceived as being inadequate so to do. As was consistently expressed by interviewees, within the competing triad of research, service delivery and teaching, the latter is perceived as taking a poor third place. CONCLUSIONS: That research, service delivery and education are in competition will come as no surprise to UK academic staff. However, our results show a striking uniformity of opinion. We would question whether existing NHS and higher education policies enable medical schools and health care organisations to deliver education of the highest quality to the enlarged student population. PMID- 19120946 TI - War as a positive medical educational experience. AB - CONTEXT: There is little empirical evidence in the literature regarding the nature of the experiences of medical students during war. In this study we set out to assess and analyse the experiences of medical students and residents in Beirut, Lebanon during the 2006 Lebanon-Israel war. METHODS: Students and residents working at the American University of Beirut Medical Center and the Beirut Government University Hospital during the July 2006 war were invited to participate. A total of 21 students or residents took part in either a focus group or one of 11 semi-structured interviews. These were recorded, transcribed and analysed to bring out common themes. RESULTS: Although the wartime experience was stressful, medical students and residents reported many largely positive effects of war on their medical training. Students adapted to the difficult conditions and became more resourceful. Participants connected emotionally with their patients and were able to show more compassion. They felt pride in their chosen profession and had higher levels of motivation for dealing with the normally irksome aspects of medical practice. Practical skills associated with the treatment of wartime injuries were learned, as were more general lessons about life priorities. CONCLUSIONS: Despite the curtailing of formal educational activities, participants benefited from positive learning experiences in a wartime environment. Strikingly, participants 'cared' more for their patients. Further research looking at the underlying cause of this increased level of compassion may be useful in the education of all medical students, not just those involved in conflict. PMID- 19120947 TI - Effect of legislative changes in drug promotion on medical students: questionnaire survey. AB - OBJECTIVE: The aim of this follow-up study was to examine whether the legislative changes that took place in Finland in 2004 had an impact on the interactions between pharmaceutical companies and medical students. According to a previous survey, information provided by pharmaceutical companies represented one of the most important sources of information on pharmaceutical products for medical students and students frequently attended promotional events. METHODS: The authors collected the survey data using questionnaires distributed to medical students in Finland's five medical departments in spring 2005. A total of 1523 students (44% of all medical students in Finland) responded to the questionnaire. Results were compared with the findings of a previous study conducted in 2000. RESULTS: We found a dramatic drop in how often students attended promotions given by pharmaceutical company representatives (PCRs), with 17% versus 68% of students in the clinical phase of study attending at least twice a month (P < 0.001). Other educational events organised by pharmaceutical companies were attended by 3% versus 22% of clinical students (P < 0.001). In addition, presentations by PCRs and industry-sponsored educational events were not regarded as such important sources of information as they had been earlier and the perceived influence of promotion on future prescribing habits had decreased (12% versus 25% indicated that promotion influences prescribing; P < 0.001). Almost two-thirds of the students indicated that basic medical education should provide them with more efficient tools for critical assessment of the claims made by pharmaceutical marketing departments. CONCLUSIONS: Legislative reform has decreased the amount of contact between the pharmaceutical industry and medical students and diminished the role of industry-sponsored promotion as a source of information on pharmaceutical products. PMID- 19120948 TI - Perceptions of learning as a function of seminar group factors. AB - CONTEXT: Small-group learning is advocated for enhancing higher-order thinking and the development of skills and attitudes. Teacher performance, group interaction and the quality of assignments have been shown to affect small-group learning in hybrid and problem-based curricula. OBJECTIVES: This study aimed to examine the perceptions of student groups as to how teacher performance, group interaction and the quality of assignments are related to one another and to learning effects in seminars of 15-30 students in a hybrid curriculum. METHODS: We constructed a 28-item questionnaire and administered it to 639 students attending 32 seminars in Years 1-4 of an undergraduate veterinary curriculum. We performed factor analysis and reliability analysis of the questionnaire. We used correlation and regression analyses to explore the interactions of the four factor model, with teacher performance, group interaction and quality of assignments as independent variables and the perceived learning effect of the seminars as the dependent variable. RESULTS: The response rate was 99%. Teacher performance (beta = 0.78) and group interaction (- 0.28) significantly influenced the perceived learning effect. The total effect of the quality of assignments (through effects on teacher performance and group interaction) was 0.47. DISCUSSION: The strong relationship between teacher performance and learning effect suggests that students rely strongly on their teachers. The negative effect of group interaction may reflect poor alignment of teaching and assessment and poor organisation of group processes. This should be further examined. Comparative studies on seminars are also recommended. PMID- 19120949 TI - 'The unknown becomes the known': collective learning and change in primary care teams. AB - CONTEXT: The growing emphasis on teamwork within the National Health Service (NHS) has made it a priority to understand how health care teams learn together and cope with change. OBJECTIVES: This study aimed to explore how collective learning and change happen in primary care teams and how the process varies across the disciplines of general medical practice, pharmacy and dentistry. METHODS: This study reports on qualitative data gathered from 10 primary care teams over 1 year, by means of observational visits and 38 semi-structured interviews. RESULTS: Informal collective learning is a powerful team coping mechanism that develops through experiential, evolving and implicit learning processes. These processes are predominantly relational in that they rely on the extent to which team members know and understand one another as people. This makes shared learning an effective but 'messy' dynamic, the motivation for which is internally generated by the team itself. Teams report that if they cannot learn together, they cannot meet patient needs. CONCLUSIONS: These findings demonstrate that teams share their knowledge because they believe it has value, not because they are driven by external incentives or are monitored. This challenges the prevailing assumption that, to be effective, interprofessional learning should be externally managed. As health care develops, it will become increasingly important to consider how to support the internal learning processes of care teams as they navigate complex organisational changes and the shared learning experiences that characterise those changes. Those who support learning and development within the NHS should therefore focus on how relational processes, as well as educational content, contribute to a team's collective learning capability and the quality of care its members provide. PMID- 19120950 TI - Using job analysis to identify core and specific competencies: implications for selection and recruitment. AB - OBJECTIVE: Modern postgraduate medical training requires both accurate and reliable selection procedures. An essential first step is to conduct detailed job analysis studies. This paper reports data on a series of job analyses to develop a competency model for three secondary care specialties (anaesthesia, obstetrics and gynaecology, and paediatrics). METHODS: Three independent job analysis studies were conducted. The content validity of the resulting competency domains was tested using a questionnaire-based study with specialty trainees (specialist registrars [SpRs]) and consultants drawn from the three specialties. Job analysis was carried out in the Yorkshire and the Humber region in the UK. The validation study was administered with additional participants from the West Midlands and Trent regions in the UK. This was an exploratory study. The outcome is a set of competency domains with data on their importance at senior house officer, SpR and consultant grade in each specialty. RESULTS: The study produced a model comprising 14 general competency domains that were common to all the three specialties. However, there were significant between-specialty differences in both definitions of domains and the ratings of importance attached to them. CONCLUSIONS: The results indicate that a wide range of attributes beyond clinical knowledge and academic achievement need to be considered in order to ensure doctors train and work within a specialty for which they have a particular aptitude. This has significant implications for developing selection criteria for specialty training. Future research should explore the content validity of these competency domains in other secondary care specialties. PMID- 19120951 TI - Pilot study of a 'RIME'-based tool for giving feedback in a multi-specialty longitudinal clerkship. AB - CONTEXT: The Reporter-Interpreter-Manager-Educator (RIME) evaluation framework is intuitive and reliable. Our preceptors' frustration with using summative tools for formative feedback and the hypothesis that the RIME vocabulary might improve students' and preceptors' experiences with feedback prompted us to develop and pilot a RIME-based feedback tool. METHODS: The tool was based on the RIME vocabulary, which has previously been used for evaluation. As interpersonal skills and professionalism are difficult areas in which to give feedback, we added these as explicit categories. We piloted the tool in a longitudinal, 5 month, multi-specialty clerkship. Preceptors completed pre- and post-introductory workshop surveys. Students completed post-workshop and post-clerkship surveys. RESULTS: Preceptors (n = 14) and students (n = 8) preferred RIME-based feedback to 'usual feedback' (previously given using end-of-clerkship evaluation forms). After the initial workshop, preceptors expected that giving feedback, including critical feedback, would be easier. After the 5-month clerkship, students reported receiving more feedback than in previous clerkships and rated feedback given using this tool more highly (P = 0.002; effect size 1.2). Students also felt it helped them understand specifically how to improve their performance (P = 0.003; effect size 1.2). DISCUSSION: In this pilot study, preceptors and students preferred feedback with a specific RIME-based tool. Students felt such feedback was more useful and helped them identify specifically how to improve. Whether this method can improve student performance through improved feedback remains an area for further research. PMID- 19120952 TI - Acceptability and educational impact of a peer feedback model for significant event analysis. AB - CONTEXT: A model of independent, external review of significant event analysis by trained peers was introduced by NHS Scotland in 1998 to support the learning needs of general practitioners (GPs). Engagement with this feedback model has increased over time, but participants' views and experiences are largely unknown and there is limited evidence of its educational impact. This is important if external feedback is to play a potential role in appraisal and future revalidation. OBJECTIVE: The study aimed to explore aspects of the acceptability and educational impact of this external feedback model with participating GPs. METHODS: Semi-structured interviews were carried out with nine GPs. Participants were sampled to reflect their level of learning need (low, moderate or high) to gain a range of views and experiences. Transcribed interviews were analysed for content. RESULTS: This system of external peer feedback is generally acceptable to participants. It complemented and enhanced the appraisal process. External feedback had positive educational outcomes, particularly in imparting technical knowledge on how to analyse significant events. Training issues for peer reviewers were suggested that would further enhance the educational gain from participation. There was disagreement over whether this type of feedback could or should be used as supporting evidence of the quality of doctors' work to educational and regulatory authorities. CONCLUSIONS: The findings add to the evidence for the acceptability and educational impact of external review by trained peers. Aligning such a model with the current national appraisal system may provide GPs with a more robust demonstration of participation in reflective learning. PMID- 19120953 TI - Understanding student concerns about peer physical examination using an activity theory framework. AB - CONTEXT: Peer physical examination (PPE) has been employed for several decades as part of the formal curriculum for learning clinical skills. Most of the existing studies exploring students' attitudes towards PPE are single-site and use quantitative methods. Currently, there is a lack of theoretical underpinning to PPE as a learning method. METHODS: Using an adaptation of the Examining Fellow Students questionnaire, we captured qualitative data from Year 1 medical students about their views and concerns around learning using PPE. The study was set in six schools across five countries (the UK, Australia, New Zealand, Japan and Hong Kong). Students provided free text comments that were later transcribed and analysed using framework analysis. RESULTS: A total of 617 students provided comments for analysis. This paper focuses on several related themes about the complexities of students' relationships within the context of PPE and their reflections on peer examination in comparison with genuine patient examination. Students drew parallels and differences between the peer examiner-examinee relationship and the doctor-patient relationship. They explained how these two types of relationship differed in nature and in terms of their levels of interaction. DISCUSSION: Our findings illuminate the interactional and complex nature of PPE, drawing out concerns and ambiguities around relationships, community and rules. We discuss our results in light of Engestrom's model of activity theory (AT) and provide recommendations for educational practice and further research based on the principles of AT. PMID- 19120954 TI - Relationship between candidate communication ability and oral certification examination scores. AB - CONTEXT: Structured case-based oral examinations are widely used in medical certifying examinations in the USA. These orals assess the candidate's decision making skills using real or realistic patient cases. Frequently mentioned but not empirically evaluated is the potential bias introduced by the candidate's communication ability. OBJECTIVE: This study aimed to assess the relationship between candidate communication ability and medical certification oral examination scores. METHODS: Non-doctor communication observers rated a random sample of 90 candidates on communication ability during a medical oral certification examination. The multi-facet Rasch model was used to analyse the communication survey and the oral examination data. The multi-facet model accounts for observer and examiner severity bias. anova was used to measure differences in communication ability between passing and failing candidates and candidates grouped by level of communication ability. Pearson's correlations were used to compare candidate communication ability and oral certification examination performance. RESULTS: Candidate separation reliability values for the communication survey and the oral examination were 0.85 and 0.97, respectively, suggesting accurate candidate measurement. The correlation between communication scores and oral examination scores was 0.10. No significant difference was found between passing and failing candidates for measured communication ability. When candidates were grouped by high, moderate and low communication ability, there was no significant difference in their oral certification examination performance. CONCLUSIONS: Candidates' communication ability has little relationship to candidate performance on high-stakes, case-based oral examinations. Examiners for this certifying examination focused on assessing candidate decision-making ability and were not influenced by candidate communication ability. PMID- 19120955 TI - Evaluation of a novel assessment form for observing medical residents: a randomised, controlled trial. AB - CONTEXT: Teaching faculty cannot reliably distinguish between satisfactory and unsatisfactory resident performances and give non-specific feedback. OBJECTIVES: This study aimed to test whether a novel rating form can improve faculty accuracy in detecting unsatisfactory performances, generate more rater observations and improve feedback quality. METHODS: Participants included two groups of 40 internal medicine residency faculty staff. Both groups received 1-hour training on how to rate trainees in the mini-clinical evaluation exercise (mini-CEX) format. The intervention group was given a new rating form structured with prompts, space for free-text comments, behavioural anchors and fewer scoring levels, whereas the control group used the current American Board of Internal Medicine Mini-CEX form. Participants watched and scored six scripted videotapes of resident performances 2-3 weeks after the training session. RESULTS: Intervention group participants were more accurate in discriminating satisfactory from unsatisfactory performances (85% versus 73% correct; odds ratio [OR] 2.13, 95% confidence interval [CI] 1.16-3.14, P = 0.02) and yielded more correctly identified unsatisfactory performances (96% versus 52% correct; OR 25.35, 95% CI 9.12-70.46), but were less accurate in identifying satisfactory performances (73% versus 95% correct; OR 0.15, 95% CI 0.05-0.39). Intervention group participants averaged one fewer declared intended feedback item (4.7 versus 5.7) and showed no difference in the amount of feedback that was above minimal in quality. Intervention group participants generated more written evaluative observations (10.8 versus 5.7). Inter-rater agreement improved with the new form (Fleiss' kappa, 0.52 versus 0.30). CONCLUSIONS: Modifying the currently used direct observations process may produce more recorded observations, increase inter-rater agreement and improve overall rater accuracy, but it may also increase severity error. PMID- 19120956 TI - Sheep blowfly strike risk and management in Great Britain: a survey of current practice. AB - The methods used for the control of sheep blowfly strike (ovine cutaneous myiasis) and the farm management factors associated with strike prevalence were examined using data from questionnaire survey returns provided by 966 sheep farmers in Great Britain, based on the period between March 2003 and February 2004. Overall, 91% of participants treated prophylactically with insecticides against blowfly strike; 39% treated twice and 11% treated more than three times in the year. Insect growth regulators (IGRs) were the most commonly chosen product (40%), especially the IGR cyromazine. Only 12% of farmers opted to dip their sheep in organophosphate insecticide against fly strike and 2% of farmers reported applying inappropriate products against strike to their sheep, such as ivermectin or 'drenches'. Farmers worming their ewes more often were 0.8 times less likely to report blowfly strike, but those who wormed their lambs more often were 1.2 times more likely to report strike. Pure-breed flocks were 0.7 times less likely to record an outbreak of blowfly strike than cross-breed flocks. Strike was less likely in ewe flocks grazed at higher altitude; however, this relationship with altitude was not seen in lambs. The results show that insecticides remain the primary tool used by almost all farmers to prevent strike and that the type of insecticides used and means of application have altered dramatically over the past 15 years. However, the prevalence of strike has remained almost unchanged over this period. Clearly careful attention to the type and timing of insecticide application, in association with a detailed understanding of the husbandry factors that predispose sheep to higher strike risk, is essential to allow the optimal management of strike problems. PMID- 19120957 TI - Ontogenetic study of three Calliphoridae of forensic importance through cuticular hydrocarbon analysis. AB - The ontogeny of the cuticular hydrocarbons of three dipterans of importance to forensic entomology, Calliphora vomitoria (Linne), Calliphora vicina (Robineau Desvoidy) and Protophormia terraenovae (Robineau-Desvoidy) (Diptera: Calliphoridae), was explored using gas chromatography analysis. The stages examined ranged from eggs to 8-day-old adults. Cuticular hydrocarbon profiles revealed that odd linear alkanes dominate in all three species. Short-chain carbon compounds in larvae and post-feeding larvae were seen to evolve to long chain carbon compounds in pupae and adults. Discriminant analysis of cuticular hydrocarbon profiles showed a clear differentiation among the different stages (larvae, post-feeding larvae, pupae and adults) and within stages, according to the age of individuals. This study concluded that the postmortem interval can be determined from the composition of cuticular waxes in Calliphoridae in forensic situations. PMID- 19120958 TI - Distribution and molecular detection of Theileria and Babesia in questing ticks from northern Spain. AB - A total of 562 questing adult ixodid ticks, collected during 2003-05 in 10 recreational mountain areas in northern Spain, were analysed for piroplasm infection. Reverse line blot (RLB) analysis using a panel of probes for 23 piroplasm species identified 16 different piroplasms, with an overall prevalence of 9.3%. Most were Theileria spp.-positive (7.7%), 3.0% were positive for Babesia spp. and 1.4% of ticks harboured both genera. Ixodes ricinus (Linnaeus, 1758), the most abundant tick in the vegetation, ranked third with regard to piroplasm infection prevalence (11.4%) after Rhipicephalus bursa (Canestrini & Fanzago, 1878) (16.0%) and Haemaphysalis punctata (Canestrini & Fanzago, 1878) (13.5%). Infection was detected in 6.2% of Dermacentor reticulatus (Fabricius, 1794) and in 1.1% of Haemaphysalis inermis (Birula, 1895), but was absent from Haemaphysalis concinna (Koch, 1844). Ixodes ricinus carried more piroplasm species (13), followed by H. punctata (10), D. reticulatus (8), R. bursa (3) and H. inermis (1). Although most of the positive ticks harboured a single infection (76.9%), mixed infections with two or three different piroplasm species were also detected (23.1%). The various tick-pathogen associations found are discussed and prevalences of infection in ticks are compared with previous results on piroplasms infecting animals in the same region. PMID- 19120959 TI - Resistance to insect growth regulator insecticides in populations of sheep lice as assessed by a moulting disruption assay. AB - Low-volume, backline applications with the benzoylphenyl urea insecticides triflumuron and diflubenzuron represent in excess of 70% of treatments for the control of sheep lice, Bovicola ovis (Schrank) (Phthiraptera: Trichodectidae), in Australia. Reports of reduced effectiveness from 2003 and subsequent controlled treatment trials suggested the emergence of resistance to these compounds in B. ovis populations. A laboratory assay based on the measurement of moulting success in nymphs was developed and used to assess susceptibility to diflubenzuron and triflumuron in louse populations collected from sheep where a control failure had occurred. These tests confirmed the development of resistance to triflumuron and diflubenzuron in at least two instances, with estimated resistance ratios of 67 94X at LC(50). PMID- 19120960 TI - Resistance to benzoylphenyl urea insecticides in Australian populations of the sheep body louse. AB - A laboratory bioassay based on the failure of eggs laid by diflubenzuron-treated sheep body lice Bovicola ovis (Schrank) (Phthiraptera; Trichodectidae) to produce nymphs is described. The test is capable of detecting benzoylphenyl urea resistant populations. A susceptible discriminating dose (topical treatment of female lice with 0.10 microg diflubenzuron) suitable for screening suspect resistant populations is suggested but testing of further populations that are naive to these insect growth regulator insecticides is warranted. PMID- 19120961 TI - Ovicidal and adulticidal effects of monoterpenoids against permethrin-resistant human head lice, Pediculus humanus capitis. AB - The improper use of pediculicides containing permethrin has led to the development of resistance. Thus, new alternatives for control are needed. Plant derived insecticides are attractive alternatives to common chemical insecticides because most of them are environmentally friendly and non-toxic to mammals. The toxic activity of 23 monoterpenoids belonging to several chemical classes was tested against the eggs of permethrin-resistant head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae). Significant differences in ovicidal action were observed among the tested substances. The most effective chemicals were hydrocarbons and ethers, followed by ketones, alcohols, phenols and esters. A linear relationship between egg mortality and knockdown time (KT(50)) on adults by the tested components revealed that most of the components were effective on both egg and adult stages. The monoterpenoids described herein are good candidates as effective pediculicides. PMID- 19120962 TI - Incursion and range expansion in the bluetongue vector Culicoides imicola in the Mediterranean basin: a phylogeographic analysis. AB - The bluetongue (BT) vector Culicoides imicola Kieffer (Diptera: Ceratopogonidae) has undergone widespread range expansion across most of the Mediterranean basin, concomitant with the largest BT epizootic outbreaks on record. Knowledge of the substructure of this vector expansion would be useful for identifying specific source-expansion systems. To this end we analysed the haplotype diversity of the mitochondrial cytochrome oxidase I gene in 273 C. imicola from 88 Mediterranean sites and outgroups. All the C. imicola haplotypes (n = 26) formed a single, distinct clade in comparison with haplotypes of four other species of the Imicola group from southern Africa, confirming C. imicola as a single phylospecies. Haplotype distribution showed extreme differentiation across the Mediterranean basin, with four common haplotypes each predominating in different areas. Eastern and western areas characterized by distinct BT incursions accounted for most of the molecular variance in haplotype composition. Shared common haplotypes identified one area of incursion and expansion encompassing the western half of the Mediterranean basin, with evidence of population growth, and another system encompassing Anatolian Turkey, the Aegean Islands and mainland Greece. A third area of range expansion was identified in the central Mediterranean, with a possible source in Algeria and unsampled parts of central North Africa. We conclude that the expansion of C. imicola in the Mediterranean basin consists of at least three incursions followed by expansions and that the western system experiences conditions promoting high population growth. PMID- 19120963 TI - Accidental importation of the mosquito Aedes albopictus into the Netherlands: a survey of mosquito distribution and the presence of dengue virus. AB - In the summer of 2005, the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae) was found for the first time in the Netherlands. It was collected on the premises of several horticultural companies that import the ornamental plant Dracaena sanderiana (Sparagalus: Dracaenaceae [Agavaceae]), known as Lucky bamboo, from southern China, an area endemic for this mosquito species and for arboviruses transmitted by this vector. Here we report the results of a 1-year survey of the distribution and vector status of Ae. albopictus in Lucky bamboo nurseries in the Netherlands (July 2006-June 2007). As it had been established previously that the presence of this species was linked to the import of Lucky bamboo, the survey was conducted only on sites owned by relevant import companies. In total, 569 adult Ae. albopictus were collected with mosquito traps from 15 of the 17 (88%) glasshouses used by Lucky bamboo importers, none of which were found to be infected with dengue virus. On two occasions there was evidence that Ae. albopictus had escaped from the glasshouses, but, overall, there was no evidence that a population had become established in the greenhouses or elsewhere. PMID- 19120964 TI - A recent survey of mosquito fauna in Guangdong Province,southern China, with a review of past records [corrected]. AB - The southern province of Guangdong has long been subject to endemic mosquito borne diseases. In recent years, this region of China has experienced rapid, extensive economic development involving environmental change, making much of the scant knowledge of its mosquito fauna obsolete. This paper reviews previous mosquito surveys, some of which may be too old to be of relevance to present-day conditions, and presents the results of a recent survey of adult and immature mosquitoes. The main vectors of mosquito-borne diseases endemic to the area, such as dengue and Japanese encephalitis virus, develop in container habitats. A three year survey was carried out, between 2004 and 2006, of 4131 breeding containers in residential areas and in open, sparsely populated areas, of which approximately 50% were positive for the presence of mosquitoes, and 10 156 larvae and pupae were collected and identified. Twelve species were found in both residential and sparsely populated areas: Aedes albopictus (Skuse) (Diptera: Culicidae), Ae. lineatopennis (Ludlow), Ae. vexans (Meigen), Tanakaius togoi (Theobald), Culex barraudi Edwards, Cx dispectus Bram, Cx malayi (Leicester), Cx pallidothorax Theobald, Cx quinquefasciatus Say, Cx sitiens Wiedemann, Lutzia fuscanus Wiedemann and Tripteroides bambusa (Yamada). Armigeres subalbatus (Coquillett) was found only in containers in villages, whereas Ae. macfarlanei Edwards, Cx mimeticus Noe, Cx sinensis Theobald, Cx vegans Wiedemann, Cx wilfredi Colless and Mansonia uniformis (Theobald) were found only in non- or sparsely populated areas. In residential areas, the rank order of most common species, as measured by the proportion of containers colonized, was Ae. albopictus > Cx quinquefasciatus > Lu. fuscanus, whereas in sparsely populated areas the rank order was Cx quinquefasciatus > Ae. albopictus > Lu. fuscanus. Light traps in non or sparsely populated areas caught 5995 adult mosquitoes of 25 species, some of which are not container breeders. The most common species were: Anopheles sinensis Wiedemann, An. maculatus Theobald, An. minimus Theobald, Ta. togoi, Cx bitaeniorhynchus Giles, Cx malayi, Cx quinquefasciatus, Cx sinensis Theobald, Cx sitiens, Cx tritaeniorhynchus Giles and Lu. fuscanus. It is noteworthy that nine species caught had not been previously recorded in Guangdong Province, highlighting the deficient knowledge of the current composition and distribution of the mosquito fauna of this part of China. PMID- 19120965 TI - Seasonal and spatial dynamics of ectoparasite infestation of a threatened reptile, the tuatara (Sphenodon punctatus). AB - The conservation of threatened vertebrate species and their threatened parasites requires an understanding of the factors influencing their distribution and dynamics. This is particularly important for species maintained in conservation reserves at high densities, where increased contact among hosts could lead to increased rates of parasitism. The tuatara (Sphenodon punctatus) (Reptilia: Sphenodontia) is a threatened reptile that persists at high densities in forests (approximately 2700 tuatara/ha) and lower densities in pastures and shrubland (< 200 tuatara/ha) on Stephens Island, New Zealand. We investigated the lifecycles and seasonal dynamics of infestation of two ectoparasites (the tuatara tick, Amblyomma sphenodonti, and trombiculid mites, Neotrombicula sp.) in a mark recapture study in three forest study plots from November 2004 to March 2007, and compared infestation levels among habitat types in March 2006. Tick loads were lowest over summer and peaked from late autumn (May) until early spring (September). Mating and engorgement of female ticks was highest over spring, and larval tick loads subsequently increased in early autumn (March). Nymphal tick loads increased in September, and adult tick loads increased in May. Our findings suggest the tuatara tick has a 2- or 3-year lifecycle. Mite loads were highest over summer and autumn, and peaked in March. Prevalences (proportion of hosts infected) and densities (estimated number of parasites per hectare) of ticks were similar among habitats, but tick loads (parasites per host) were higher in pastures than in forests and shrub. The prevalence and density of mites was higher in forests than in pasture or shrub, but mite loads were similar among habitats. We suggest that a higher density of tuatara in forests may reduce the ectoparasite loads of individuals through a dilution effect. Understanding host parasite dynamics will help in the conservation management of both the host and its parasites. PMID- 19120966 TI - Identification of mosquito bloodmeals using mitochondrial cytochrome oxidase subunit I and cytochrome b gene sequences. AB - Primer pairs were designed and protocols developed to selectively amplify segments of vertebrate mitochondrial cytochrome oxidase subunit 1 (COI) and cytochrome b (Cyt b) mtDNA from the bloodmeals of mosquitoes (Diptera: Culicidae). The protocols use two pairs of nested COI primers and one pair of Cyt b primers to amplify short segments of DNA. Resultant sequences are then compared with sequences in GenBank, using the BLAST function, for putative host identification. Vertebrate DNA was amplified from 88% of our sample of 162 wild caught, blood-fed mosquitoes from Oregon, U.S.A. and GenBank BLAST searches putatively identified 98% of the amplified sequences, including one amphibian, seven mammalian and 14 avian species. Criteria and caveats for putative identification of bloodmeals are discussed. PMID- 19120967 TI - Identification of immune-responsive genes in the mosquito Culex quinquefasciatus infected with the filarial parasite Wuchereria bancrofti. AB - Several antimicrobial/parasitic peptides are known to be upregulated in mosquitoes upon infection with parasites. The aim of this study was to identify immune-responsive genes in the vector mosquito, Culex quinquefasciatus Say (Diptera: Culicidae) against the human lymphatic filarial parasite, Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae). Suppression subtractive hybridization was performed using RNA from filarial infected and non-infected mosquitoes to obtain differentially expressed transcripts, and their identities were confirmed through reverse transcriptase polymerase chain reaction (RT-PCR). Out of 23 clones selected from the suppression subtractive library, three corresponded to antimicrobial peptide genes, defensins, and four corresponded to regulatory serpin peptide genes. RT-PCR using defensin-specific primers and sequencing of the product showed a 284-bp defensin cDNA. Sequence alignment with defensins of the mosquitoes Anopheles gambiae s.s. Giles and Aedes aegypti (L.) showed maximum homology with the former. Similarly, that of serpin-specific primers showed a 406-bp cDNA encoding serpins. Sequence alignment showed maximum homology with that of An. gambiae, as in the case of defensins. Hence, this investigation revealed upregulation of defensins and serpins in Cx quinquefasciatus infected with W. bancrofti. Antimicrobial peptide genes such as defensins may have limited or no specific role in regulating parasite development. Serpins may prove to be facilitating molecules, by regulating melanization of the parasite. However, the exact functions of these molecules in the immune system of the vector mosquito are yet to be investigated. PMID- 19120968 TI - The effect of predatory fish exudates on the ovipostional behaviour of three mosquito species: Culex quinquefasciatus, Aedes aegypti and Culex tarsalis. AB - Three mosquito species, Culex tarsalis Coquillett, Culex quinquefasciatus Say and Aedes aegypti L. (Diptera: Culicidae), were examined in laboratory binary choice experiments to investigate whether fish exudates from the mosquitofish, Gambusia affinis (Baird & Girard) (Cyprinodontiformes: Poecilliidae), deter oviposition and whether the responses of these mosquito species to fish exudates in oviposition sites are consistent with the risk of predation from fish experienced by each species in their respective natural breeding habitats. Culex tarsalis was deterred significantly from egg laying by the presence of fish exudates in oviposition cups, consistent with high levels of predation by fish in natural breeding sites. Egg laying by Cx quinquefasciatus was slightly reduced in water with fish exudates, but was not consistently deterred by water conditioned by mosquitofish, consistent with the species' relatively low risk of fish predation in natural habitats. Oviposition by container-breeding Ae. aegypti was not deterred by the presence of fish exudates in oviposition cups, consistent with a low risk of predation by fish in natural habitats. PMID- 19120969 TI - Control of phlebotomine sandflies in confined spaces using diffusible repellents and insecticides. AB - The control of phlebotomine sandflies (Diptera: Psychodidae), the vectors of leishmaniasis, is directed mostly against adults as larvae develop in unknown or inaccessible habitats. In the current study we tested geraniol, a natural plant derived product, as a space repellent and the synthetic pyrethroid prallethrin as a diffusible insecticide. Geraniol was dispersed in the air using diffusers with an electric fan and prallethrin was evaporated using electrically heated evaporators. Both substances were tested in inhabited bedrooms and in tents. Geraniol failed to effect significant reductions in the numbers of either Phlebotomus papatasi Scopoli in rooms or Phlebotomus sergenti Parrot in tents. In laboratory experiments, geraniol proved ineffective in preventing sandflies from feeding. By contrast, prallethrin was highly effective in reducing the number of sandflies in rooms as well as in tents. Exposure of sandflies to prallethrin in laboratory experiments caused 97% mortality rates. Both prallethrin and, to a lesser extent, geraniol reduced the number of Culex mosquitoes captured in tents. Electric liquid-vaporizers with 1.5% prallethrin are highly effective in protecting people from sandfly bites in confined spaces and may be useful in combating cutaneous leishmaniasis. PMID- 19120970 TI - Regulation of marA, soxS, rob, acrAB and micF in Salmonella enterica serovar Typhimurium. AB - Importance of the overexpression of AcrAB efflux pumps in the low-level resistance of pathogens to antimicrobials requires a better understanding of the AcrAB regulation. The goal of the present research was to study the transcription of acrAB, as well as the genes that play a role in its regulation in Salmonella enterica serovar Typhimurium. We monitored the transcription of these genes during growth at 30 degrees C and 37 degrees C, and thoroughly studied the effect of salicylate, paraquat and decanoate. The strengths of the promoters' activities were ordered from strong to weak as micF > rob > acrAB > soxS, marRAB. At both temperatures, marRAB was mainly upregulated by salicylate and decanoate, soxS by paraquat and acrAB and micF by all three compounds. rob was always downregulated. Transcription rates of all promoters were at the greatest level at the beginning of the stationary phase and, except for soxS, levels of transcription and induction were greater at 37 degrees C. Results show that despite the promoters' similar activity and the sequence homology between Escherichia coli and S. typhimurium, regulation of the investigated genes of both strains differed in the response to temperature. This difference was found to be dependent on the promoters' sequence, as the marRAB and acrAB promoters maintained their original dependence on temperature when they were analyzed in the other strain. Hence, it is most likely that the nutrient-limited environment at 37 degrees C in the human body will lead to increased transcription of marA, acrAB and micF in Salmonella, enhancing the resistance properties of the bacteria. PMID- 19120971 TI - Herpes simplex virus type 1 dysregulates anti-fungal defenses preventing monocyte activation and downregulating toll-like receptor-2. AB - We investigated the interplay occurring between pathogens in the course of dual infections, using an in vitro model in which the THP-1 monocytic cell line is first infected with HSV-1 and then exposed to Ca or Cn. These three pathogens share some pathogenic features: they cause opportunistic infections, target macrophages and are neurotropic. Here, we show that HSV-1-infected THP-1 cells exhibited augmented phagocytosis against the two opportunistic fungi but reduced capability to counteract fungal infection: the better ingestion by monocytes was followed by facilitated fungal survival and replication. Reduced IL-12 production was also observed. Cytofluorimetric analysis showed that HSV-1-infected monocytes exhibit: (i) downregulated TLR-2 and TLR-4, critical structures in fungal recognition; (ii) reduced expression of CD38 and CD69, known to be important markers of monocyte activation; and (iii) enhanced expression of apoptosis and necrosis markers, in the absence of altered cell proliferation. Overall, these findings imply that HSV-1 infection prevents monocyte activation, thus leading to a significant dysfunction of the monocyte-mediated anti-Candida response; HSV-1 induced apoptosis and necrosis of monocytes further contribute to this impairment. PMID- 19120972 TI - Receptor activator of nuclear factor-kappa B ligand induces osteoclast formation in RAW 264.7 macrophage cells via augmented production of macrophage-colony stimulating factor. AB - RAW 264.7 macrophage cells differentiate into osteoclast-like cells in the presence of RANKL. Participation of M-CSF in RANKL-induced osteoclast formation of RAW 264.7 cells was examined. TRAP-positive osteoclast-like cells appeared in RAW 264.7 cells cultured in the presence of RANKL. RANKL-induced osteoclast formation was markedly inhibited by anti-M-CSF antibody. RANKL augmented M-CSF mRNA expression and M-CSF production in RAW 264.7 cells. Further, anti-M-CSF antibody inhibited the expression of RANK, c-fms, c-fos and TRAP mRNA in RANKL stimulated RAW 264.7 cells. However, anti-M-CSF antibody did not affect the expression of DC-STAMP in the stimulated cells. Therefore, RANKL was suggested to induce osteoclast formation in RAW 264.7 cells via augmented production of M-CSF. The putative role of M-CSF in RANKL-induced osteoclast formation of RAW 264.7 cells is discussed. PMID- 19120973 TI - A WT1 protein-derived, naturally processed 16-mer peptide, WT1(332), is a promiscuous helper peptide for induction of WT1-specific Th1-type CD4(+) T cells. AB - The Wilms' tumor gene WT1 is overexpressed in various tumors, and the WT1 protein has been demonstrated to be an attractive target antigen for cancer immunotherapy. A WT1 protein-derived 16-mer peptide, WT1(332) (KRYFKLSHLQMHSRKH), which was naturally generated through processing in cells and could elicit Th1 type CD4(+) helper T cell responses with an HLA-DRB1*0405-restriction has previously been identified by us. In the present study, it has been demonstrated that WT1(332) can induce WT1(332)-specific CD4(+) T cell responses with the restriction of not only HLA-DRB1*0405 but also HLA-DRB1*1501, -DRB1*1502, or DPB1*0901. These HLA class II-restricted WT1(332)-specific CD4(+) T cell lines produced IFN-gamma but neither IL-4 nor IL-10 with WT1(332) stimulation, thus showing a Th1-type cytokine profile. Furthermore, HLA-DRB1*1501 or -DRB1*1502 restricted WT1(332)-specific CD4(+) T cell lines responded to WT1-expressing transformed cells in an HLA-DRB1-restricted manner, which is consistent with our previous finding that WT1(332) is a naturally processed peptide. These results indicate that the natural peptide, WT1(332), is a promiscuous WT1-specific helper epitope. WT1(332) is expected to apply to cancer patients with various types of HLA class II as a WT1-specific helper peptide in combination with HLA class I restricted WT1 peptides. PMID- 19120974 TI - Experimental autoimmune uveoretinitis initiated by non-phagocytic destruction of inner segments of photoreceptor cells by Mac-1(+) mononuclear cells. AB - EAU in mice is a model of human posterior uveitis. EAU is a Th1-dependent disease that has been assumed to target the neural retina and related tissues; however, in situ effector cells and the target have not yet been clearly demonstrated. In the present study, we induced EAU in B10R mice by immunizing them with human interphotoreceptor retinoid-binding protein peptide 161-180. Histological examinations revealed that EAU occurred approximately 11 days after the immunization and reached a peak on day 14. Retinae from normal or EAU mice were treated with proteases to obtain mono-dispersed cells. The mono-dispersed cells thus obtained were separated into three to four fractions by discontinuous Percoll density-gradient (e.g. PBS/40/60) centrifugation. In normal mice, 94% of the total cells were recovered in two fractions (i.e. PBS/40 and pellet); and these fractions mainly contained inner and outer segments and cell bodies of photoreceptor cells and RPE cells, respectively. In EAU mice, additional cells (i.e. inflammatory cells) were obtained at the 40/60 interface. Electron microscopic examination showed that tissue damage during EAU was initiated by non phagocytic destruction of inner segments by Mac-1(+) mononuclear cells on day 11, followed by phagocytic activity of macrophages against outer segments and RPE cells on day 14. In vitro culturing of normal retinal cells with EAU infiltrates suggested the involvement of TNF-alpha and NO in the tissue damage. These results indicate that EAU was initiated by non-phagocytic destruction of inner segments of photoreceptor cells by Mac-1(+) mononuclear cells. PMID- 19120975 TI - Variable gene family usage of protective and non-protective anti-Vibrio cholerae O1 LPS antibody heavy chains. AB - Vibrio cholerae causes cholera, an enteric disease of humans that is a worldwide problem. The O1 serogroup of Vibrio cholerae contains two predominant serotypes (Inaba and Ogawa) of LPS, a proven protective antigen for humans and experimental animals. We generated B-cell hybridomas from mice immunized with either: (i) two doses of purified Inaba LPS; (ii) two doses of an Inaba hexasaccharide conjugate (terminal six perosamine bound to a protein carrier), (iii) four doses of purified Inaba LPS; or (iv) a low dose of purified Inaba LPS followed by a booster with the Inaba conjugate. We showed previously that the first and third immunization protocols induce vibriocidal antibodies, as does the fourth; the second protocol induces antibodies that bind Inaba and Ogawa LPS but are not vibriocidal. Anti-LPS mAbs derived from hybridomas resulting from each immunization protocol were characterized for binding to Inaba and Ogawa LPS, their vibriocidal or protective capacity, and the variable heavy chain family they expressed. LPS immunogens selected different LPS-specific B cells expressing six different Vh chain families. Protective and non-protective mAbs could express variable regions from the same family. One mAb was specific for Inaba LPS, the other mAbs were cross-reactive with both LPS serotypes. Sequence comparison suggests that the pairing of a specific light chain, somatic mutation, or the specific VDJ recombination can modulate the protective capacity of mAbs that express a common variable heavy chain family member. PMID- 19120976 TI - Antimicrobial susceptibilities of Propionibacterium acnes isolated from patients with acne vulgaris. AB - Antibiotic susceptibilities of Propionibacterium acnes in Japan were determined. Erythromycin-resistance was found in 10.4% (5/48) of the strains, and four of these were cross-resistance to clindamycin. Although the erythromycin ribosome methylase gene erm(X) was looked for, no strain carrying erm(X) was found. Sequencing analysis revealed that all of the erythromycin-resistant strains had a mutation in the peptidyl transferase region of the 23S rRNA gene: G2057A, A2058G, or A2059G. Consequently, our results show that P. acnes resistance to macrolides is caused by a mutation in the 23S rRNA gene, and has been increasing in Japan. PMID- 19120977 TI - Analysis of severe acute respiratory syndrome coronavirus structural proteins in virus-like particle assembly. AB - SARS-CoV has four major structural proteins: the N, S, M, and E proteins. To investigate the mechanism of SARS-CoV assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pCAGGS and transfected them into 293T cells. When all four expression vectors were co-transfected VLP formed, as confirmed using electron microscopy. Using a rabbit polyclonal antibody specific to the N protein, N-protein-containing particles similar in size to the VLP were also observed by immunoelectron microscopy, indicating that the VLP contained the N protein. Co-immunoprecipitation analyses demonstrated an interaction between the N and M proteins, suggesting that N protein binds directly to M protein to be incorporated into VLP. PMID- 19120978 TI - Milbank quarterly. PMID- 19120979 TI - What ever happened to N-of-1 trials? Insiders' perspectives and a look to the future. AB - CONTEXT: When feasible, randomized, blinded single-patient (n-of-1) trials are uniquely capable of establishing the best treatment in an individual patient. Despite early enthusiasm, by the turn of the twenty-first century, few academic centers were conducting n-of-1 trials on a regular basis. METHODS: The authors reviewed the literature and conducted in-depth telephone interviews with leaders in the n-of-1 trial movement. FINDINGS: N-of-1 trials can improve care by increasing therapeutic precision. However, they have not been widely adopted, in part because physicians do not sufficiently value the reduction in uncertainty they yield weighed against the inconvenience they impose. Limited evidence suggests that patients may be receptive to n-of-1 trials once they understand the benefits. CONCLUSIONS: N-of-1 trials offer a unique opportunity to individualize clinical care and enrich clinical research. While ongoing changes in drug discovery, manufacture, and marketing may ultimately spur pharmaceutical makers and health care payers to support n-of-1 trials, at present the most promising resuscitation strategy is stripping n-of-1 trials to their essentials and marketing them directly to patients. In order to optimize statistical inference from these trials, empirical Bayes methods can be used to combine individual patient data with aggregate data from comparable patients. PMID- 19120980 TI - The relative merits of population-based and targeted prevention strategies. AB - CONTEXT: Preventive medicine has historically favored reducing a risk factor by a small amount in the entire population rather than by a large amount in high-risk individuals. The use of multivariable risk prediction tools, however, may affect the relative merits of this strategy. METHODS: This study uses risk factor data from the National Health and Nutrition Examination Survey III to simulate a population of more than 100 million Americans aged thirty or older with no history of CV disease. Three strategies that could affect CV events, CV mortality, and quality-adjusted life years were examined: (1) a population-based strategy that treats all individuals with a low- or moderate-intensity intervention (in which the low-intensity intervention represents a public health campaign with no demonstrable adverse effects), (2) a targeted strategy that treats individuals in the top 25 percent based on a single risk factor (LDL), and (3) a risk-targeted strategy that treats individuals in the top 25 percent based on overall CV risk (as predicted by a multivariable prediction tool). The efficiency of each strategy was compared while varying the intervention's intensity and associated adverse effects, and the accuracy of the risk prediction tool. FINDINGS: The LDL-targeted strategy and the low-intensity population-based strategy were comparable for CV events prevented over five years (0.79 million and 0.75 million, respectively), as were the risk-targeted strategy and moderate intensity population-based strategy (1.56 million and 1.87 million, respectively). The risk-targeted strategy, however, was more efficient than the moderate-intensity population-based strategy (number needed to treat [NNT] 19 vs. 62). Incorporating a small degree of treatment-related adverse effects greatly magnified the relative advantages of the risk-targeted approach over other strategies. Reducing the accuracy of the prediction tool only modestly decreased this greater efficiency. CONCLUSIONS: A population-based prevention strategy can be an excellent option if an intervention has almost no adverse effects. But if the intervention has even a small degree of disutility, a targeted approach using multivariable risk prediction can prevent more morbidity and mortality while treating many fewer people. PMID- 19120981 TI - Revisiting rose: comparing the benefits and costs of population-wide and targeted interventions. AB - CONTEXT: Geoffrey Rose's two principal approaches to public health intervention are (1) targeted strategies focusing on individuals at a personal increased risk of disease and (2) population-wide approaches focusing on the whole population. Beyond his discussion of the strengths and weaknesses of these approaches, there is no empiric work examining the conditions under which one of these approaches may be better than the other. METHODS: This article uses mathematical simulations to model the benefits and costs of the two approaches, varying the cut points for treatment, effect magnitudes, and costs of the interventions. These techniques then were applied to the specific example of an intervention on blood pressure to reduce cardiovascular disease. FINDINGS: In the general simulation (using an inverse logit risk curve), lower costs of intervention, treating people with risk factor values at or above where the slope on the risk curve is at its steepest (for targeted interventions), and interventions with larger effects on reducing the risk factor (for population-wide interventions) provided benefit/cost advantages. In the specific blood pressure intervention example, lower-cost population-wide interventions had better benefit/cost ratios, but some targeted treatments with lower cutoffs prevented more absolute cases of disease. CONCLUSIONS: These simulations empirically evaluate some of Rose's original arguments. They can be replicated for particular interventions being considered and may be useful in helping public health decision makers assess potential intervention strategies. PMID- 19120982 TI - Helping smokers quit: understanding the barriers to utilization of smoking cessation services. AB - CONTEXT: Counseling smokers to quit smoking and providing them with pharmaceutical cessation aides are among the most beneficial and cost-effective interventions that clinicians can offer patients. Yet assistance with quitting is not universally covered by health plans or offered by all clinicians. Analysis of stakeholders' perspectives and interests can identify the barriers to more widespread provision of cessation services and suggest strategies for the public policy agenda to advance smoking cessation. METHODS: Review of literature and discussions with representatives of stakeholders. FINDINGS: All stakeholders health plans, employers, clinicians, smokers, and the government-face barriers to broader smoking cessation activities. These range from health plans' perceiving that covering counseling and pharmacotherapy will increase costs without producing commensurate health care savings, to clinicians' feeling unprepared and uncompensated for counseling. Like other preventive measures aimed at behavior, efforts directed at smoking cessation have marginal status among health care interventions. State governments can help correct this status by increasing Medicaid coverage of treatment and expanding coverage for state employees. The federal government can promote the adoption of six initiatives recommended by a government subcommittee on cessation: set up a national quit line, develop a media campaign to encourage cessation, include cessation benefits in all federally funded insurance plans, create a research infrastructure to improve cessation rates, develop a clinician training agenda, and create a fund to increase cessation activities through a new $2 per pack cigarette excise tax. Both the federal and state governments can increase cessation by adopting policies such as the higher cigarette tax and laws prohibiting smoking in workplaces and public places. CONCLUSIONS: Public policy efforts should assume greater social responsibility for smoking cessation, including more aggressive leadership at the state and federal levels, as well as through advocacy, public health, and clinician organizations. PMID- 19120984 TI - Revealing the hidden complexities of mtDNA inheritance. AB - Mitochondrial DNA (mtDNA) is a pivotal tool in molecular ecology, evolutionary and population genetics. The power of mtDNA analyses derives from a relatively high mutation rate and the apparent simplicity of mitochondrial inheritance (maternal, without recombination), which has simplified modelling population history compared to the analysis of nuclear DNA. However, in biology things are seldom simple, and advances in DNA sequencing and polymorphism detection technology have documented a growing list of exceptions to the central tenets of mitochondrial inheritance, with paternal leakage, heteroplasmy and recombination now all documented in multiple systems. The presence of paternal leakage, recombination and heteroplasmy can have substantial impact on analyses based on mtDNA, affecting phylogenetic and population genetic analyses, estimates of the coalescent and the myriad of other parameters that are dependent on such estimates. Here, we review our understanding of mtDNA inheritance, discuss how recent findings mean that established ideas may need to be re-evaluated, and we assess the implications of these new-found complications for molecular ecologists who have relied for decades on the assumption of a simpler mode of inheritance. We show how it is possible to account for recombination and heteroplasmy in evolutionary and population analyses, but that accurate estimates of the frequencies of biparental inheritance and recombination are needed. We also suggest how nonclonal inheritance of mtDNA could be exploited, to increase the ways in which mtDNA can be used in analyses. PMID- 19120983 TI - Waste in the U.S. Health care system: a conceptual framework. AB - CONTEXT: Health care costs in the United States are much higher than those in industrial countries with similar or better health system performance. Wasteful spending has many undesirable consequences that could be alleviated through waste reduction. This article proposes a conceptual framework to guide researchers and policymakers in evaluating waste, implementing waste-reduction strategies, and reducing the burden of unnecessary health care spending. METHODS: This article divides health care waste into administrative, operational, and clinical waste and provides an overview of each. It explains how researchers have used both high level and sector- or procedure-specific comparisons to quantify such waste, and it discusses examples and challenges in both waste measurement and waste reduction. FINDINGS: Waste is caused by factors such as health insurance and medical uncertainties that encourage the production of inefficient and low-value services. Various efforts to reduce such waste have encountered challenges, such as the high costs of initial investment, unintended administrative complexities, and trade-offs among patients', payers', and providers' interests. While categorizing waste may help identify and measure general types and sources of waste, successful reduction strategies must integrate the administrative, operational, and clinical components of care, and proceed by identifying goals, changing systemic incentives, and making specific process improvements. CONCLUSIONS: Classifying, identifying, and measuring waste elucidate its causes, clarify systemic goals, and specify potential health care reforms that-by improving the market for health insurance and health care-will generate incentives for better efficiency and thus ultimately decrease waste in the U.S. health care system. PMID- 19120985 TI - Population genetics of Galapagos land iguana (genus Conolophus) remnant populations. AB - The Galapagos land iguanas (genus Conolophus) have faced significant anthropogenic disturbances since the 17th century, leading to severe reduction of some populations and the extinction of others. Conservation activities, including the repatriation of captive-bred animals to depleted areas, have been ongoing since the late 1970s, but genetic information has not been extensively incorporated. Here we use nine species-specific microsatellite loci of 703 land iguanas from the six islands where the species occur today to characterize the genetic diversity within, and the levels of genetic differentiation among, current populations as well as test previous hypotheses about accidental translocations associated with early conservation efforts. Our analyses indicate that (i) five populations of iguanas represent distinct conservation units (one of them being the recently discovered rosada form) and could warrant species status, (ii) some individuals from North Seymour previously assumed to be from the natural Baltra population appear related to both Isabela and Santa Cruz populations, and (iii) the five different management units exhibit considerably different levels of intrapopulation genetic diversity, with the Plaza Sur and Santa Fe populations particularly low. Although the initial captive breeding programmes, coupled with intensive efforts to eradicate introduced species, saved several land iguana populations from extinction, our molecular results provide objective data for improving continuing in situ species survival plans and population management for this spectacular and emblematic reptile. PMID- 19120986 TI - Genetic and acoustic population structuring in the Okinawa least horseshoe bat: are intercolony acoustic differences maintained by vertical maternal transmission? AB - The origin and meaning of echolocation call frequency variation within rhinolophid bats is not well understood despite an increasing number of allopatric and sympatric examples being documented. A bimodal distribution of mean regional call frequency within the Okinawa-jima Island population of Rhinolophus cornutus pumilus (Rhinolophidae) provided a unique opportunity to investigate geographic call frequency variation early in its development. Individual resting echolocation frequencies, partial mitochondrial DNA D-loop sequences and genotypes from six microsatellite loci were obtained from 288 individuals in 11 colonies across the entire length of the island, and nearby Kume-jima Island. Acoustic differences (5-8 kHz) observed between the north and south regions have been maintained despite evidence of sufficient nuclear gene flow across the middle of the island. Significant subdivision of maternally inherited D-loop haplotypes suggested a limitation of movement of females between regions, but not within the regions, and was evidence of female philopatry. These results support a 'maternal transmission' hypothesis whereby the difference in the constant frequency (CF) component between the regions is maintained by mother offspring transmission of CF, the restricted dispersal of females between regions and small effective population size. We suggest that the mean 5-8 kHz call frequency difference between the regions might develop through random cultural drift. PMID- 19120987 TI - Genetic patterns across multiple introductions of the globally invasive crab genus Carcinus. AB - The European green crab Carcinus maenas is one of the world's most successful aquatic invaders, having established populations on every continent with temperate shores. Here we describe patterns of genetic diversity across both the native and introduced ranges of C. maenas and its sister species, C. aestuarii, including all known non-native populations. The global data set includes sequences from the mitochondrial cytochrome c oxidase subunit I gene, as well as multilocus genotype data from nine polymorphic nuclear microsatellite loci. Combined phylogeographic and population genetic analyses clarify the global colonization history of C. maenas, providing evidence of multiple invasions to Atlantic North America and South Africa, secondary invasions to the northeastern Pacific, Tasmania, and Argentina, and a strong likelihood of C. maenas x C. aestuarii hybrids in South Africa and Japan. Successful C. maenas invasions vary broadly in the degree to which they retain genetic diversity, although populations with the least variation typically derive from secondary invasions or from introductions that occurred more than 100 years ago. PMID- 19120988 TI - Drastic post-LGM expansion and lack of historical genetic structure of a subtropical fig-pollinating wasp (Ceratosolen sp. 1) of Ficus septica in Taiwan. AB - The climatic oscillations of the last glacial period have had profound influences on the demography and levels of genetic diversity of extant species. Molecular evidence of glacial effects on temperate species has been well documented, whereas little is known regarding that on subtropical species. Here we present analyses based on partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene (1052 bp) and genotypes at 15 microsatellite loci to investigate the historical demography, population structure and ongoing gene flow of an undescribed fig-pollinating wasp (Ceratosolen sp. 1) of Ficus septica in subtropical Taiwan. Reconstructed historical demography based on the coalescent tree of COI sequences suggests that C. sp. 1 has undergone a drastic population expansion which was tightly coupled with climatic changes since the last glacial maximum (LGM). The magnitude of the population size change was approximately 500 fold, indicating that the population of this wasp and its host was likely highly compressed during the last glacial period. The lack of significant population differentiation (F(ST) < 0.02, for all pairwise F(ST) values) may be due to rapid postglacial expansion facilitated by long-distance dispersal, although a low frequency of first-generation migrants was detected. Our results clearly demonstrate how recent climatic changes since the LGM and dispersal ability have jointly shaped the genetic composition of a subtropical fig-pollinating wasp. PMID- 19120989 TI - Dispersal in the spiny damselfish, Acanthochromis polyacanthus, a coral reef fish species without a larval pelagic stage. AB - The spiny damselfish, Acanthochromis polyacanthus, is widely distributed throughout the Indo-Australian archipelago. However, this species lacks a larval dispersal stage and shows genetic differentiation between populations from closely spaced reefs. To investigate the dispersal strategy of this unique species, we used microsatellite markers to determine genetic relatedness at five dispersal scales: within broods of juveniles, between adults within a collection site (approximately 30 m(2)), between sites on single reefs, between nearby reefs in a reef cluster, and between reef clusters. We sampled broods of juveniles and adults from seven reefs in the Capricorn-Bunker and Swain groups of the Great Barrier Reef. We found that extra-pair mating is rare and juveniles remain with their parents until fledged. Adults from single sites are less related than broods but more related than expected by chance. However, there is no evidence of inbreeding suggesting the existence of assortative mating and/or adult migration. Genetic differences were found between all of the reefs tested except between Heron and Sykes reefs, which are separated only by a 2-km area of shallow water (less than 10 m). There was a strong correlation between genetic distance, geographical distance and water depth. Apparently, under present-day conditions spiny damselfish populations are connected only between sites of shallow water, through dispersal of adults over short distances. Assuming that dispersal behaviour has not changed, the broad distribution of A. polyacanthus as a species is likely based on historical colonization patterns when reefs were connected by shallow water at times of lower sea levels. PMID- 19120990 TI - Polyploid evolution and plastid DNA variation in the Dactylorhiza incarnata/maculata complex (Orchidaceae) in Scandinavia. AB - The Dactylorhiza incarnata/maculata complex (Orchidaceae) was used as a model system to understand genetic differentiation processes in a naturally occurring polyploid complex with much of ongoing diversification and wide distribution in recently glaciated areas in northern Europe. Data were obtained for 12 hypervariable regions in the plastid DNA genome. A total of 166 haplotypes were found in a sample of 1099 plants. Allopolyploid taxa have inherited their plastid genomes from D. maculata s.l. Overall haplotype diversity of the combined group of allopolyploid taxa was comparable to that of maternal D. maculata s.l., but populations of allopolyploids were also more strongly differentiated from each other and contained lower numbers of haplotypes than populations of D. maculata s.l. In addition to haplotypes found in extant D. maculata s.l., the allopolyploids also contained several distinct and widespread haplotypes that were not found in any of the parental lineages. Some of these haplotypes were shared between widespread allopolyploids. Divergent allopolyploids with small distributions did not seem to originate from local polyploidization events, but rather as segregates of already existing allopolyploids. Genetic diversification of allopolyploid Dactylorhiza is the result of repeated polyploid formation, secondary hybridization and introgression between already existing polyploids and extant representatives of parental lineages, hybridization between independently derived polyploid lineages, and phyletic diversification in the group of allopolyploids. Although some polyploid taxa must have evolved after the last glaciation, genetic material from the parental lineages has been transferred continuously for longer periods of time. This combination of processes may explain the taxonomic complexity encountered in Dactylorhiza and other polyploid complexes distributed in previously glaciated parts of Europe. PMID- 19120991 TI - Airborne-pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla. AB - The reproductive isolation barriers and the mating patterns among Pinus pumila, P. parviflora var. pentaphylla and their hybrids were examined by flowering phenology and genetic assays of three life stages: airborne-pollen grains, adults and seeds, in a hybrid zone on Mount Apoi, Hokkaido, Japan. Chloroplast DNA composition of the airborne-pollen was determined by single-pollen polymerase chain reaction. Mating patterns were analysed by estimating the molecular hybrid index of the seed parent, their seed embryos and pollen parents. The observation of flowering phenology showed that the flowering of P. pumila precedes that of P. parviflora var. pentaphylla by about 6 to 10 days within the same altitudinal ranges. Although this prezygotic isolation barrier is effective, the genetic assay of airborne-pollen showed that the two pine species, particularly P. pumila, still have chances to form F(1) hybrid seeds. Both parental species showed a strong assortative mating pattern; F(1) seeds were found in only 1.4% of seeds from P. pumila mother trees and not at all in P. parviflora var. pentaphylla. The assortative mating was concluded as the combined result of flowering time differentiation and cross-incompatibility. In contrast to the parental species, hybrids were fertilized evenly by the two parental species and themselves. The breakdown of prezygotic barriers (intermediate flowering phenology) and cross-incompatibility may account for the unselective mating. It is suggested that introgression is ongoing on Mount Apoi through backcrossing between hybrids and parental species, despite strong isolation barriers between the parental species. PMID- 19120992 TI - Comparative phylogeography of two sibling species of forest-dwelling rodent (Praomys rostratus and P. tullbergi) in West Africa: different reactions to past forest fragmentation. AB - Two sibling species of the rodent genus Praomys occur in West African forests: P. tullbergi and P. rostratus. By sampling across their geographical ranges (459 individuals from 77 localities), we test the hypothesis that climatic oscillations during the Quaternary made an impact on the observed pattern of cytochrome b sequence variation. We show that, although these two species have parapatric geographical distributions, their phylogeographical histories are dissimilar, which could be related to their distinct ecological requirements. Since the arid phases of the Pleistocene were characterized by isolated forest patches, and intervening wetter periods by forest expansion, these changes in forest cover may be the common mechanism responsible for the observed phylogeographical patterns in both of these species. For example, in both species, most clades had either allopatric or parapatric geographical distributions; however, genetic diversity was much lower in P. tullbergi than in P. rostratus. The genetic pattern of P. tullbergi fits the refuge hypothesis, indicating that a very small number of populations survived in distinct forest blocks during the arid phases, then expanded again with forest recovery. In contrast, a number of populations of P. rostratus appear to have survived during the dry periods in more fragmented forest habitats, with varying levels of gene flow between these patches depending on climatic conditions and forest extent. In addition, historical variations of the West African hydrographic network could also have contributed to the pattern of genetic differentiation observed in both species. PMID- 19120993 TI - The roles of allopatric divergence and natural selection in quantitative trait variation across a secondary contact zone in the lizard Anolis roquet. AB - Populations of the Caribbean lizard, Anolis roquet, are thought to have experienced long periods of allopatry before recent secondary contact. To elucidate the effects of past allopatry on population divergence in A. roquet, we surveyed parallel transects across a secondary contact zone in northeastern Martinique. We used diagnostic molecular mitochondrial DNA markers to test fine scale association of mitochondrial DNA lineage and geological region, multivariate statistical techniques to explore quantitative trait pattern, and cline fitting techniques to model trait variation across the zone of secondary contact. We found that lineages were strongly associated with geological regions along both transects, but quantitative trait patterns were remarkably different. Patterns of morphological and mitochondrial DNA variation were consistent with a strong barrier to gene flow on the coast, whereas there were no indications of barriers to gene flow in the transitional forest. Hence, the coastal populations behaved as would be predicted by an allopatric model of divergence in this complex, while those in the transitional forest did not, despite the close proximity of the transects and their shared geological history. Patterns of geographical variation in this species complex, together with environmental data, suggest that on balance, selection regimes on either side of the secondary contact zone in the transitional forest may be more convergent, while those either side of the secondary contact zone on the coast are more divergent. Hence, the evolutionary consequences of allopatry may be strongly influenced by local natural selection regimes. PMID- 19120994 TI - Using patterns of genetic structure based on microsatellite loci to test hypotheses of current hybridization, ancient hybridization and incomplete lineage sorting in Conradina (Lamiaceae). AB - Hybridization and/or incomplete sorting of ancestral polymorphism are commonly implicated to explain discordant phylogenetic analyses of closely related species complexes. One genus in which these phenomena have been suggested to have played major roles based on phylogenetic data is Conradina, a genus of mints (Lamiaceae) endemic to the southeastern USA containing several endangered species. The goals of this study were to use microsatellite data to better understand patterns of genetic structure in Conradina, to test hypotheses of recent or ancient hybridization and incomplete lineage sorting, and to clarify species boundaries. Individuals from 55 populations representing all Conradina species were genotyped using 10 microsatellite loci. Analyses of the patterns of genetic structure in Conradina revealed a clear differentiation of populations following recognized species boundaries, indicating that species have diverged from one another genetically and interspecific hybridization has not occurred recently. Neither ancient hybridization nor incomplete lineage sorting is supported as the sole cause of species nonmonophyly, suggesting that both may have contributed to patterns found in phylogenetic trees; however, analyses of other types of data may be more appropriate to distinguish between these two hypotheses. Because all described species appear to be valid entities, the current listing status of most endangered species of Conradina is appropriate; however, populations of Conradina canescens are genetically differentiated into three groups, each of which may merit species status, and several recently discovered populations of Conradina in Dunn's Creek State Park in Florida are highly differentiated genetically and also appear to represent a new species. PMID- 19120995 TI - Genetic consequences of habitat fragmentation in plant populations: susceptible signals in plant traits and methodological approaches. AB - Conservation of genetic diversity, one of the three main forms of biodiversity, is a fundamental concern in conservation biology as it provides the raw material for evolutionary change and thus the potential to adapt to changing environments. By means of meta-analyses, we tested the generality of the hypotheses that habitat fragmentation affects genetic diversity of plant populations and that certain life history and ecological traits of plants can determine differential susceptibility to genetic erosion in fragmented habitats. Additionally, we assessed whether certain methodological approaches used by authors influence the ability to detect fragmentation effects on plant genetic diversity. We found overall large and negative effects of fragmentation on genetic diversity and outcrossing rates but no effects on inbreeding coefficients. Significant increases in inbreeding coefficient in fragmented habitats were only observed in studies analyzing progenies. The mating system and the rarity status of plants explained the highest proportion of variation in the effect sizes among species. The age of the fragment was also decisive in explaining variability among effect sizes: the larger the number of generations elapsed in fragmentation conditions, the larger the negative magnitude of effect sizes on heterozygosity. Our results also suggest that fragmentation is shifting mating patterns towards increased selfing. We conclude that current conservation efforts in fragmented habitats should be focused on common or recently rare species and mainly outcrossing species and outline important issues that need to be addressed in future research on this area. PMID- 19120996 TI - Polymorphic MHC loci in an asexual fish, the amazon molly (Poecilia formosa; Poeciliidae). AB - Genes of the major histocompatibility complex (MHC) encode molecules that control immune recognition and are highly polymorphic in most vertebrates. The remarkable polymorphisms at MHC loci may be maintained by selection from parasites, sexual selection, or both. If asexual species show equal (or higher) levels of polymorphisms at MHC loci as sexual ones, this would mean that sexual selection is not necessary to explain the high levels of diversity at MHC loci. In this study, we surveyed the MHC diversity of the asexual amazon molly (Poecilia formosa) and one of its sexual ancestors, the sailfin molly (P. latipinna), which lives in the same habitat. We found that the asexual molly has polymorphic MHC loci despite its clonal reproduction, yet not as polymorphic as the sexual species. Although the nucleotide diversity was similar between the asexual and sexual species, the sexual species exhibited a greater genotypic diversity compared to the asexual one from the same habitats. Within-genome diversity was similar for MHC class I loci, but for class IIB, the sexual species had higher diversity compared to the asexual--despite the hybrid origins and higher levels of heterozygosity at microsatellite loci in the asexual species. The level of positive selection appears to be similar between the two species, which suggests that these polymorphisms are maintained by selection. Thus, our findings do not allow us to rule out the sexual selection hypothesis for the evolution of MHC diversity, and although the sexual fish has higher levels of MHC-diversity compared to the asexual species, this may be due to differences in demography, parasites, or other factors, rather than sexual selection. PMID- 19120997 TI - Widespread mito-nuclear discordance with evidence for introgressive hybridization and selective sweeps in Lycaeides. AB - We investigated the extent and potential cause(s) of mitochondrial introgression within the polytypic North American Lycaeides species complex (Lepidoptera). By comparing population genetic structure based on mitochondrial DNA (COI and COII) and nuclear DNA (251 polymorphic amplified fragment length polymorphism markers), we detected substantial mito-nuclear discordance, primarily involving a single mitochondrial haplotype (h01), which is likely due to mitochondrial introgression between differentiated Lycaeides populations and/or species. We detected reduced mitochondrial genetic diversity relative to nuclear genetic diversity in populations where mitochondrial haplotype h01 occurs, suggesting that the spread of this haplotype was facilitated by selection. We found no evidence that haplotype h01 is associated with increased fitness (in terms of survival to eclosion, fresh adult weight, and adult longevity) in a polymorphic Lycaeides melissa population. However, we did find a positive association between mitochondrial haplotype h01 and infection by the endoparasitic bacterium Wolbachia in one out of three lineages tested. Linkage disequilibrium between mitochondrial haplotype h01 and Wolbachia infection status may have resulted in indirect selection favouring the spread of haplotype h01 in at least one lineage of North American Lycaeides. These results illustrate the potential for introgressive hybridization to produce substantial mito-nuclear discordance and demonstrate that an individual's mitochondrial and nuclear genome may have strikingly different evolutionary histories resulting from non-neutral processes and intrinsic differences in the inheritance and biology of these genomes. PMID- 19120998 TI - The evolution of sexual and parthenogenetic Warramaba: a window onto Plio Pleistocene diversification processes in an arid biome. AB - Environmental changes over the Plio-Pleistocene have been key drivers of speciation patterns and genetic diversification in high-latitude and mesic environments, yet comparatively little is known about the evolutionary history of species in arid environments. We applied phylogenetic and phylogeographic analyses to understand the evolutionary history of Warramaba grasshoppers from the Australian arid zone, a group including sexual and parthenogenetic lineages. Sequence data (mitochondrial COI) showed that the four major sexual lineages within Warramaba most likely diverged in the Pliocene, around 2-7 million years ago. All sexual lineages exhibited considerable phylogenetic structure. Detailed analyses of the hybrid parthenogenetic species W. virgo and its sexual progenitors showed a pattern of high phylogenetic diversity and phylogeographic structure in northern lineages, and low diversity and evidence for recent expansion in southern lineages. Northern sexual lineages persisted in localized refugia over the Pleistocene, with sustained barriers promoting divergence over this period. Southern parts of the present range became periodically unsuitable during the Pleistocene, and it is into this region that parthenogenetic lineages have expanded. Our results strongly parallel those for sexual and parthenogenetic lineages of the gecko Heteronotia from the same region, indicating a highly general effect of Plio-Pleistocene environmental change on diversification processes in arid Australia. PMID- 19120999 TI - Sea-level changes and palaeo-ranges: reconstruction of ancient shorelines and river drainages and the phylogeography of the Australian land crayfish Engaeus sericatus Clark (Decapoda: Parastacidae). AB - Historical sea levels have been influential in shaping the phylogeography of freshwater-limited taxa via palaeodrainage and palaeoshoreline connections. In this study, we demonstrate an approach to phylogeographic analysis incorporating historical sea-level information in a nested clade phylogeographic analysis (NCPA) framework, using burrowing freshwater crayfish as the model organism. Our study area focuses on the Bass Strait region of southeastern Australia, which is marine region encompassing a shallow seabed that has emerged as a land bridge during glacial cycles connecting mainland Australia and Tasmania. Bathymetric data were analysed using Geographical Information Systems (GIS) to delineate a palaeodrainage model when the palaeocoastline was 150 m below present-day sea level. Such sea levels occurred at least twice in the past 500 000 years, perhaps more often or of larger magnitude within the last 10 million years, linking Victoria and Tasmania. Inter-locality distance measures confined to the palaeodrainage network were incorporated into an NCPA of crayfish (Engaeus sericatus Clark 1936) mitochondrial 16S rDNA haplotypes. The results were then compared to NCPAs using present-day river drainages and traditional great-circle distance measures. NCPA inferences were cross-examined using frequentist and Bayesian procedures in the context of geomorphological and historical sea-level data. We found distribution of present-day genetic variation in E. sericatus to be partly explained not only by connectivity through palaeodrainages but also via present-day drainages or overland (great circle) routes. We recommend that future studies consider all three of these distance measures, especially for studies of coastally distributed species. PMID- 19121000 TI - Lineage diversification and historical demography of a sky island salamander, Plethodon ouachitae, from the Interior Highlands. AB - Sky islands provide ideal opportunities for understanding how climatic changes associated with Pleistocene glacial cycles influenced species distributions, genetic diversification, and demography. The salamander Plethodon ouachitae is largely restricted to high-elevation, mesic forest on six major mountains in the Ouachita Mountains. Because these mountains are separated by more xeric, low elevation valleys, the salamanders appear to be isolated on sky islands where gene flow among populations on different mountains may be restricted. We used DNA sequence data along with ecological niche modelling and coalescent simulations to test several hypotheses related to diversifications in sky island habitats. Our results revealed that P. ouachitae is composed of seven well-supported lineages structured across six major mountains. The species originated during the Late Pliocene, and lineage diversification occurred during the Middle Pleistocene in a stepping stone fashion with a cyclical pattern of dispersal to a new mountain followed by isolation and divergence. Diversification occurred primarily on an east-west axis, which is likely related to the east-west orientation of the Ouachita Mountains and the more favourable cooler and wetter environmental conditions on north slopes compared to south-facing slopes and valleys. All non genealogical coalescent methods failed to detect significant population expansion in any lineages. Bayesian skyline plots showed relatively stable population sizes over time, but indicated a slight to moderate amount of population growth in all lineages starting approximately 10 000-12 000 years ago. Our results provide new insight into sky island diversifications from a previously unstudied region, and further demonstrate that climatic changes during the Pleistocene had profound effects on lineage diversification and demography, especially in species from environmentally sensitive habitats in montane regions. PMID- 19121001 TI - Global phylogeography and seascape genetics of the lemon sharks (genus Negaprion). AB - Seascapes are complex environments, and populations are often isolated by factors other than distance. Here we investigate the role of coastal habitat preference and philopatry in shaping the distribution and population structure of lemon sharks. The genus Negaprion comprises the amphiatlantic lemon shark (N. brevirostris), with a relict population in the eastern Pacific, and its Indo-West Pacific sister species, the sicklefin lemon shark (N. acutidens). Analyzing 138 individuals throughout the range of N. brevirostris (N = 80) and N. acutidens (N = 58) at microsatellite loci (nine and six loci, respectively) and the mitochondrial control region, we find evidence of allopatric speciation corresponding to the Tethys Sea closure (10-14 million years ago) and isolation of the eastern Pacific N. brevirostris population via the emergence of the Isthmus of Panama (approximately 3.5 million years ago). There is significant isolation by oceanic distance (R(2) = 0.89, P = 0.005), defined as the maximum distance travelled at depths greater than 200 m. We find no evidence for contemporary transatlantic gene flow (m, M = 0.00) across an oceanic distance of approximately 2400 km. Negaprion acutidens populations in Australia and French Polynesia, separated by oceanic distances of at least 750 km, are moderately differentiated (F(ST) = 0.070-0.087, P < or = 0.001; Phi(ST) = 0.00, P = 0.99), with South Pacific archipelagos probably serving as stepping stones for rare dispersal events. Migration between coastally linked N. brevirostris populations is indicated by nuclear (m = 0.31) but not mitochondrial (m < 0.001) analyses, possibly indicating female natal site fidelity. However, philopatry is equivocal in N. acutidens, which has the lowest control region diversity (h = 0.28) of any shark yet studied. Restricted oceanic dispersal and high coastal connectivity stress the importance of both local and international conservation efforts for these threatened sharks. PMID- 19121003 TI - Recent divergence, intercontinental dispersal and shared polymorphism are shaping the genetic structure of amphi-Atlantic peatmoss populations. AB - Several lines of evidence suggest that recent long-distance dispersal may have been important in the evolution of intercontinental distribution ranges of bryophytes. However, the absolute rate of intercontinental migration and its relative role in the development of certain distribution ranges is still poorly understood. To this end, the genetic structure of intercontinental populations of six peatmoss species showing an amphi-Atlantic distribution was investigated using microsatellite markers. Methods relying on the coalescent were applied (IM and MIGRATE) to understand the evolution of this distribution pattern in peatmosses. Intercontinental populations of the six peatmoss species were weakly albeit significantly differentiated (average F(ST) = 0.104). This suggests that the North Atlantic Ocean is acting as a barrier to gene flow even in bryophytes adapted to long-range dispersal. The im analysis suggested a relatively recent split of intercontinental populations dating back to the last two glacial periods (9000-289,000 years ago). In contrast to previous hypotheses, analyses indicated that both ongoing migration and ancestral polymorphism are important in explaining the intercontinental genetic similarity of peatmoss populations, but their relative contribution varies with species. Migration rates were significantly asymmetric towards America suggesting differential extinction of genotypes on the two continents or invasion of the American continent by European lineages. These results indicate that low genetic divergence of amphi-Atlantic populations is a general pattern across numerous flowering plants and bryophytes. However, in bryophytes, ongoing intercontinental gene flow and retained shared ancestral polymorphism must both be considered to explain the genetic similarity of intercontinental populations. PMID- 19121002 TI - Inferring the history of speciation in house mice from autosomal, X-linked, Y linked and mitochondrial genes. AB - Patterns of genetic differentiation among taxa at early stages of divergence provide an opportunity to make inferences about the history of speciation. Here, we conduct a survey of DNA-sequence polymorphism and divergence at loci on the autosomes, X chromosome, Y chromosome and mitochondrial DNA in samples of Mus domesticus, M. musculus and M. castaneus. We analyzed our data under a divergence with gene flow model and estimate that the effective population size of M. castaneus is 200,000-400,000, of M. domesticus is 100,000-200,000 and of M. musculus is 60,000-120,000. These data also suggest that these species started to diverge approximately 500,000 years ago. Consistent with this recent divergence, we observed considerable variation in the genealogical patterns among loci. For some loci, all alleles within each species formed a monophyletic group, while at other loci, species were intermingled on the phylogeny of alleles. This intermingling probably reflects both incomplete lineage sorting and gene flow after divergence. Likelihood ratio tests rejected a strict allopatric model with no gene flow in comparisons between each pair of species. Gene flow was asymmetric: no gene flow was detected into M. domesticus, while significant gene flow was detected into both M. castaneus and M. musculus. Finally, most of the gene flow occurred at autosomal loci, resulting in a significantly higher ratio of fixed differences to polymorphisms at the X and Y chromosomes relative to autosomes in some comparisons, or just the X chromosome in others, emphasizing the important role of the sex chromosomes in general and the X chromosome in particular in speciation. PMID- 19121004 TI - Population-genomic approach reveals adaptive floral divergence in discrete populations of a hawk moth-pollinated violet. AB - Local adaptation to contrasting biotic or abiotic environments is an important evolutionary step that presumably precedes floral diversification at the species level, yet few studies have demonstrated the adaptive nature of intraspecific floral divergence in wild plant populations. We combine a population-genomic approach with phenotypic information on floral traits to examine whether the differentiation in metric floral traits exhibited by 14 populations of the southern Spanish hawk moth-pollinated violet Viola cazorlensis reflects adaptive divergence. Screening of many amplified fragment length polymorphism (AFLP) loci using a multiple-marker-based neutrality test identified nine outlier loci (2.6% of the total) that departed from neutral expectations and were potentially under selection. Generalized analysis of molecular variance revealed significant relationships between genetic distance and population divergence in three floral traits when genetic distance was based on outlier loci, but not when it was based on neutral ones. Population means of floral traits were closely correlated with population scores on the first principal coordinate axis of the genetic distance matrix using outlier loci, and with the allelic frequencies of four of the outlier loci. Results strongly support the adaptive nature of intraspecific floral divergence exhibited by V. cazorlensis and illustrate the potential of genome scans to identify instances of adaptive divergence when used in combination with phenotypic information. PMID- 19121005 TI - Small membrane proteins found by comparative genomics and ribosome binding site models. AB - The correct annotation of genes encoding the smallest proteins is one of the biggest challenges of genome annotation, and perhaps more importantly, few annotated short open reading frames have been confirmed to correspond to synthesized proteins. We used sequence conservation and ribosome binding site models to predict genes encoding small proteins, defined as having 16-50 amino acids, in the intergenic regions of the Escherichia coli genome. We tested expression of these predicted as well as previously annotated genes by integrating the sequential peptide affinity tag directly upstream of the stop codon on the chromosome and assaying for synthesis using immunoblot assays. This approach confirmed that 20 previously annotated and 18 newly discovered proteins of 16-50 amino acids are synthesized. We summarize the properties of these small proteins; remarkably more than half of the proteins are predicted to be single transmembrane proteins, nine of which we show co-fractionate with cell membranes. PMID- 19121019 TI - How valid are the results of the St. Petersburg-USA orphanage intervention study and what do they mean for the world's children? PMID- 19121020 TI - Institutional effects on children: design issues and substantive findings. PMID- 19121007 TI - The effects of early social-emotional and relationship experience on the development of young orphanage children. The St. Petersburg-USA Orphanage Research Team. PMID- 19121021 TI - Earlier is better: a meta-analysis of 70 years of intervention improving cognitive development in institutionalized children. PMID- 19121025 TI - Stable carbon isotopes reveal dynamics of respiratory metabolism. PMID- 19121026 TI - Spores of ectomycorrhizal fungi: ecological strategies for germination and dormancy. PMID- 19121027 TI - Fungal proteins in the extra-radical phase of arbuscular mycorrhiza: a shotgun proteomic picture. PMID- 19121028 TI - Shaping the calcium signature. AB - In numerous plant signal transduction pathways, Ca2+ is a versatile second messenger which controls the activation of many downstream actions in response to various stimuli. There is strong evidence to indicate that information encoded within these stimulus-induced Ca2+ oscillations can provide signalling specificity. Such Ca2+ signals, or 'Ca2+ signatures', are generated in the cytosol, and in noncytosolic locations including the nucleus and chloroplast, through the coordinated action of Ca2+ influx and efflux pathways. An increased understanding of the functions and regulation of these various Ca2+ transporters has improved our appreciation of the role these transporters play in specifically shaping the Ca2+ signatures. Here we review the evidence which indicates that Ca2+ channel, Ca2+-ATPase and Ca2+ exchanger isoforms can indeed modulate specific Ca2+ signatures in response to an individual signal. PMID- 19121029 TI - Allometry and stoichiometry of unicellular, colonial and multicellular phytoplankton. AB - Phytoplankton life forms, including unicells, colonies, pseudocolonies, and multicellular organisms, span a huge size range. The smallest unicells are less than 1 microm3 (e.g. cyanobacteria), while large unicellular diatoms may attain 10(9) microm3, being visible to the naked eye. Phytoplankton includes chemo organotrophic unicells, colonies and multicellular organisms that depend on symbionts or kleptoplastids for their capacity to photosynthesize. Analyses of physical (transport within cells, diffusion boundary layers, package effect, turgor, and vertical movements) and biotic (grazing, viruses and other parasitoids) factors indicate potential ecological constraints and opportunities that differ among the life forms. There are also variations among life forms in elemental stoichiometry and in allometric relations between biovolume and specific growth. While many of these factors probably have ecological and evolutionary significance, work is needed to establish those that are most important, warranting explicit description in models. Other factors setting limitations on growth rate (selecting slow-growing species) await elucidation. PMID- 19121030 TI - Modular gene expression in Poplar: a multilayer network approach. AB - By applying a multilayer network approach to an extensive set of Poplar microarray data, a genome-wide coexpression network has been detected and explored. Multilayer networks were generated from minimum spanning trees (MSTs) using Kruskal's algorithm from random jack-knife resamplings of half of the full data set. The final network is obtained from the union of all the generated MSTs. The gene expression correlations display a highly clustered topology, which is more pronounced when introducing links appearing in relatively few of the generated MSTs. The network also reveals a modular architecture, reflecting functional groups with relatively frequent gene-to-gene communication. Furthermore, the observed modular structure overlaps with different gene activities in different tissues, and closely related tissues show similar over- and/or under-expression patterns at the modular scale. It is shown that including links that appear in a few of the generated MSTs increases the information quality of the network. In other words, a link may be 'weak' because it reflects rare signaling events rather than merely a signal weakened by noise. The method allows, from comparisons of random 'null networks', tuning to maximize the information obtainable. PMID- 19121031 TI - Why does elevated CO2 affect time of flowering? An exploratory study using the photoperiodic flowering mutants of Arabidopsis thaliana. AB - Evidence is accumulating that the effect of CO(2) on time of flowering involves interactions with photoperiod, but the basis for this interaction is unclear. Here, which components of the photoperiod flowering pathway account for this interaction in Arabidopsis thaliana were examined. Ten mutants deficient in particular loci in the photoperiod pathway, as well as the wild type, were grown under short and long days at either ambient or elevated CO(2). Leaf number at flowering and the number of days required for induction of flowering were determined. Elevated CO(2) interacted with both the photoreceptors and the subsequent transduction reactions in the photoperiod pathway. The direction and magnitude of the effects varied with photoperiod. Elevated CO(2) also affected flowering by increasing rate of leaf production. The net effect of elevated CO(2) on time of flowering varies because CO(2) has a complex array of effects on different elements of the developmental pathway leading to flower induction that may either hasten or delay flowering depending upon the influence of other environmental factors such as photoperiod. PMID- 19121032 TI - Role of phi cells and the endodermis under salt stress in Brassica oleracea. AB - Phi cell layers were discovered in the 19th century in a small number of species, including members of the Brassicaceae family. A mechanical role was first suggested for this structure; however, this has never been demonstrated. The main objective of the present work was to analyse the ultrastructure of phi cells, their influence on ion movement from the cortex to the stele, and their contribution to salt stress tolerance in Brassica oleracea. Transmission electron microscopy and X-ray microanalysis studies were used to analyse the subcellular structure and distribution of ions in phi cells and the endodermis under salt stress. Ion movement was analysed using lanthanum as an apoplastic tracer. The ultrastructural results confirm that phi cells are specialized cells showing cell wall ingrowths in the inner tangential cell walls. X-ray microanalysis confirmed a build-up of sodium. Phi thickenings were lignified and lanthanum moved periplasmically at this level. To the best of our knowledge, this is the first study reporting the possible role of the phi cells as a barrier controlling the movement of ions from the cortex to the stele. Therefore, the phi cell layer and endodermis seem to be regulating ion transport in Brassica oleracea under salt stress. PMID- 19121033 TI - Hydraulic conductance and K+ transport into the xylem depend on radial volume flow, rather than on xylem pressure, in roots of intact, transpiring maize seedlings. AB - The investigation of transport processes in roots has so far been hampered by a lack of adequate methods to study water and solute transport simultaneously in intact, transpiring plants. The role of xylem tension in regulating volume flow and nutrient transport could not be addressed properly. In order to overcome limitations of conventional, massive-invasive methods, a gravimetric technique was used to measure water uptake by maize roots while simultaneously recording xylem pressure and xylem K(+) activity in individual xylem vessels by means of a K(+)-selective xylem probe. This minimal-invasive approach allowed the calculation of the radial K(+)flux into the root xylem and the radial root hydraulic conductance on transpiring seedlings. By changing the light regime or the osmotic pressure of the external solution, radial water and K(+) flux could be varied in order to study the interaction between water and solute transport. A major finding was that both radial K(+) transport and hydraulic conductance strongly depended on radial volume flow, whereas xylem pressure had little (if any) effect on these parameters. Results are discussed with respect to relevant membrane transport processes and their regulation by volume flow. PMID- 19121034 TI - On the metabolic origin of the carbon isotope composition of CO2 evolved from darkened light-acclimated leaves in Ricinus communis. AB - The (13)C isotopic signature (delta(13)C) of CO(2) respired from plants is widely used to assess carbon fluxes and ecosystem functioning. There is, however, a lack of knowledge of the metabolic basis of the delta(13)C value of respired CO(2). To elucidate the physiological mechanisms driving (12)C/(13)C fractionation during respiration, the delta(13)C of respired CO(2) from dark-acclimated leaves during the night, from darkened leaves during the light period, and from stems and roots of Ricinus communis was analysed. The delta(13)C of potential respiratory substrates, the respiratory quotient and the activities of phosphoenolpyruvatecarboxylase (PEPc) and key respiratory enzymes were also measured. It is shown here that the CO(2) evolved from darkened light-acclimated leaves during the light period is (13)C-enriched, and that this correlates with malate accumulation in the light and rapid malate decarboxylation just after the onset of darkness. Whilst CO(2) evolved from leaves was generally (13)C-enriched (but to a lesser extent during the night), CO(2) evolved from stems and roots was depleted compared with the putative respiratory substrates; the difference was mainly caused by intensive PEPc-catalysed CO(2) refixation in stems and roots. These results provide a physiological explanation for short-term variations of delta(13)C in CO(2), illustrating the effects of variations of metabolic fluxes through different biochemical pathways. PMID- 19121035 TI - Pronounced differences in diurnal variation of carbon isotope composition of leaf respired CO2 among functional groups. AB - The first broad species survey of diurnal variation in carbon (C) isotope signatures of leaf dark-respired CO(2) (delta(13)C(res)) is presented here and functional differences and diurnal dynamics are linked to fractionation in different respiratory pathways, based on (13)C-labelling experiments. delta(13)C(res) was analysed with a rapid in-tube incubation technique in 16 species. A large diurnal increase in delta(13)C(res) (4-8 per thousand) occurred in evergreen, slow-growing and aromatic species and correlated significantly with cumulative photosynthesis, whereas no variation occurred in herbaceous, fast growing plants or temperate trees. The diurnal increase in delta(13)C(res) declined almost proportionally to reductions in cumulative light and was reduced in growing compared with mature leaves. Pyruvate positional labelling provided direct evidence that functional groups differ in C allocation between respiratory pathways owing to different metabolic demands for growth, maintenance and secondary metabolism. Diurnal increase in C flux through pyruvate dehydrogenase (for investment in, for example, isoprene or aromatic compounds) combined with consistently low Krebs cycle activity resulted in pronounced increase in delta(13)C(res) in evergreen and aromatic species. By contrast, fast growing herbs with high respiratory demand exhibited no diurnal changes since C was fully respired. Hence, diurnal delta(13)C(res) pattern may provide information for C allocation in plants. PMID- 19121036 TI - Rhizosphere bacteria containing 1-aminocyclopropane-1-carboxylate deaminase increase yield of plants grown in drying soil via both local and systemic hormone signalling. AB - Decreased soil water availability can stimulate production of the plant hormone ethylene and inhibit plant growth. Strategies aimed at decreasing stress ethylene evolution might attenuate its negative effects. An environmentally benign (nonchemical) method of modifying crop ethylene relations - soil inoculation with a natural root-associated bacterium Variovorax paradoxus 5C-2 (containing the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase that degrades the ethylene precursor ACC), was assessed with pea (Pisum sativum) plants grown in drying soil. Inoculation with V. paradoxus 5C-2, but not with a transposome mutant with massively decreased ACC deaminase activity, improved growth, yield and water-use efficiency of droughted peas. Systemic effects of V. paradoxus 5C-2 included an amplified soil drying-induced increase of xylem abscisic acid (ABA) concentration, but an attenuated soil drying-induced increase of xylem ACC concentration. A local bacterial effect was increased nodulation by symbiotic nitrogen-fixing bacteria, which prevented a drought-induced decrease in nodulation and seed nitrogen content. Successfully deploying a single bacterial gene in the rhizosphere increased yield and nutritive value of plants grown in drying soil, via both local and systemic hormone signalling. Such bacteria may provide an easily realized, economic means of sustaining crop yields and using irrigation water more efficiently in dryland agriculture. PMID- 19121037 TI - Dual purpose secondary compounds: phytotoxin of Centaurea diffusa also facilitates nutrient uptake. AB - Traits that allow more efficient foraging for a deficient resource could increase the competitiveness of a species in resource-poor habitats. Considering the metal nutrient mobilization ability of many allelochemicals, it is hypothesized that, along with the reported toxic effect on the neighbors, these compounds could be directly involved in resource acquisition by the allelopathic plant. Using nutrient manipulation treatments in hydroponic culture, this hypothesis was tested using Centaurea diffusa, an invasive species that produces the putative phytotoxin 8-hydroxyquinoline (8HQ). The exudation of 8HQ by C. diffusa was very limited and transient. It was further shown that: C. diffusa utilizes 8HQ for its own acquisition of iron, a nutrient deficient in many of its alkaline, invaded habitats; there possibly exists a unique mechanism for the uptake of the 8HQ complexed iron (Fe) in C. diffusa, which is novel to the nongraminaceous species; although phytotoxic at very low concentrations, the toxic effect of 8HQ showed a conditional response in the presence of metals, and was significantly reduced when 8HQ was complexed with copper (Cu) and Fe. This study, in addition to elucidating one of the possible adaptive mechanisms conferring competitive advantage to C. diffusa, also outlines measures to negate the phytotoxicity of its putative allelochemical. The results indicate that the exudation of 8HQ by C. diffusa could be primarily for nutrient acquisition. PMID- 19121038 TI - The plant pathogenic fungus Gaeumannomyces graminis var. tritici improves bacterial growth and triggers early gene regulations in the biocontrol strain Pseudomonas fluorescens Pf29Arp. AB - In soil, some antagonistic rhizobacteria contribute to reduce root diseases caused by phytopathogenic fungi. Direct modes of action of these bacteria have been largely explored; however, commensal interaction also takes place between these microorganisms and little is known about the influence of filamentous fungi on bacteria. An in vitro confrontation bioassay between the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) and the biocontrol bacterial strain Pseudomonas fluorescens Pf29Arp was set up to analyse bacterial transcriptional changes induced by the fungal mycelium at three time-points of the interaction before cell contact and up until contact. For this, a Pf29Arp shotgun DNA microarray was constructed. Specifity of Ggt effect was assessed in comparison with one of two other filamentous fungi, Laccaria bicolor and Magnaporthe grisea. During a commensal interaction, Ggt increased the growth rate of Pf29Arp. Before contact, Ggt induced bacterial genes involved in mycelium colonization. At contact, genes encoding protein of stress response and a patatin-like protein were up-regulated. Among all the bacterial genes identified, xseB was specifically up-regulated at contact by Ggt but down-regulated by the other fungi. Data showed that the bacterium sensed the presence of the fungus early, but the main gene alteration occurred during bacterial-fungal cell contact. PMID- 19121039 TI - A pH signaling mechanism involved in the spatial distribution of calcium and anion fluxes in ectomycorrhizal roots. AB - Mycorrhization is a typical example of a host-pathogen symbiotic interaction where the pathogen cell biology and the host immune response coevolved several functional links. Here, the role played by ion fluxes across the root concerning nutrient uptake, osmoregulation, growth and signaling events is addressed. An ion selective vibrating probe system was used to determine the net fluxes of protons (H(+)), calcium (Ca(2+)) and anions (A(-)) along nonmycorrhizal and ectomycorrhizal (ECM) roots of Eucalyptus globulus colonized by Pisolithus sp. These data show that, from five root zones analyzed, the main effect of fungal colonization was localized to the elongation zone. Here, strong changes in ion dynamics and rhizosphere acidification capacity were observed. Additionally, ion fluxes exhibited periodic fluctuations. To verify whether these fluctuations corresponded to sustained oscillations, continuous wavelet time spectrum analysis was applied and it was determined that H(+) and A(-) fluxes from ECM roots had longer periods than nonmycorrhizal roots. By contrast, Ca(2+) oscillations were completely abolished following fungal interaction. These results are interpreted in the light of a working model in which nutrient uptake and stimulation of growth are mediated by ECM fungi and may be pH-dependent. Furthermore, the variations detected in ECM roots for H(+) and A(-) fluxes suggest a main contribution from the plant, while the results obtained for Ca(2+) point to a significant involvement of the fungus. PMID- 19121040 TI - Inoculum potential of Rhizopogon spores increases with time over the first 4 yr of a 99-yr spore burial experiment. AB - In disturbed or pioneer settings, spores and sclerotia of ectomycorrhizal fungi serve as the necessary inoculum for establishment of ectomycorrhizal-dependent trees. Yet, little is known about the persistence of these propagules through time. Here, live field soil was inoculated with known quantities of basidiospores from four pine-associated species of Rhizopogon; these samples were then buried in retrievable containers, and pine seedling bioassays of serially diluted spore samples were used to measure spore viability. In the first 4 yr, no evidence of loss of spore viability was found in the four Rhizopogon species tested, but all four species exhibited dormancy in which a maximum of 1-8% of their spores were initially receptive to pine roots. There were some differences between species in overall inoculum potential of their spores, but all species broke dormancy at a statistically similar rate. This result provides evidence for spore dormancy in a common ectomycorrhizal genus, but it also precludes our ability to estimate the longevity of the spores accurately. Nevertheless these results, coupled with the observed patterns of Rhizopogon spore banks, suggest that at least decade-long durations are likely. As this experiment progresses, the true longevity of the spores will eventually be revealed. PMID- 19121041 TI - An outlook on the fungal internal transcribed spacer sequences in GenBank and the introduction of a web-based tool for the exploration of fungal diversity. AB - The environmental and distributional data associated with fungal internal transcribed spacer (ITS) sequences in GenBank are investigated and a new web based tool with which these sequences can be explored is introduced. All fungal ITS sequences in GenBank were classified as either identified to species level or insufficiently identified and compared using BLAST. The results are made available as a biweekly updated web service that can be queried to retrieve all insufficiently identified sequences (IIS) associated with any fungal genus. The most commonly available annotation items in GenBank are isolation source (55%); country of origin (50%); and specific host (38%). The molecular sampling of fungi shows a bias towards North America, Europe, China, and Japan whereas vast geographical areas remain effectively unexplored. Mycorrhizal and parasitic genera are on average associated with more IIS than are saprophytic taxa. Glomus, Alternaria, and Tomentella are the genera represented by the highest number of insufficiently identified ITS sequences in GenBank. The web service presented (http://andromeda.botany.gu.se/emerencia.html#genus_search) offers new means, particularly for mycorrhizal and plant pathogenic fungi, to examine the IIS in GenBank in a taxon-oriented framework and to explore their metadata in an easily accessible and time-efficient manner. PMID- 19121042 TI - Physiological differences among two Penstemon species and their hybrids in field and common garden environments. AB - Hybrids can exhibit unique combinations of the physiological traits of their parents. These particular combinations may influence hybrid fitness and the evolutionary trajectory of a hybrid zone. Here, a hybrid zone between Penstemon newberryi and Penstemon davidsonii along an elevational gradient was examined, and physiological traits of parents and hybrids were measured in their native environment and a common garden. Gas exchange rates of nine different crosses were also measured. Alpine P. davidsonii had less negative pre-dawn water potential and lower water use efficiency (WUE) than its montane relative P. newberryi in a common garden and in field measurements. The species difference in WUE was attributable to lower conductance in P. newberryi in the field, but to a higher photosynthetic rate in this species in the common garden. The alpine species took less time to produce mature fruits and reached maximum photosynthetic rate at a lower temperature. Natural hybrids were intermediate for most characters. F(1) hybrids had lower conductance than progeny of natural hybrids. The intermediate WUE of natural hybrids may be one factor that allows them to persist in intermediate environments. Comparisons of different crosses suggest that the genotypic composition of hybrids influences their physiological performance. PMID- 19121043 TI - The consequences of monoecy and protogyny for mating in wind-pollinated Carex. AB - Monoecy and protogyny are widespread in wind-pollinated plants and have been interpreted as outcrossing mechanisms, though few studies have investigated their function. Carex, a large genus of anemophilous herbs, is predominantly monoecious and many species are protogynous. We investigated whether monoecy and protogyny limit self-pollination in seven Carex species. We conducted field experiments comparing stigmatic pollen loads and seed set between intact and emasculated stems. We tested for self-compatibility and evaluated pollen limitation of seed set by supplemental pollination. Finally, we measured outcrossing rates in open pollinated and emasculated stems using allozyme markers. Emasculated stems captured significantly less pollen than open-pollinated stems and set less seed. Pollen deposition during the female-only phase for intact stems was only 12% of the total captured. Outcrossing rates for three species indicated high selfing (range t = 0.03-0.39). Allozyme loci in the remaining species were monomorphic also suggesting high selfing. These results demonstrate that neither monoecy nor protogyny is particularly effective at limiting self-fertilization. Selection for the avoidance of selfing is unlikely to maintain monoecy in many Carex species although protogyny may provide limited opportunities for outcrossing. We propose that geitonogamy in self-compatible wind-pollinated species with unisexual flowers may be widespread and provides reproductive assurance. PMID- 19121045 TI - I say mentor, you say mentee. PMID- 19121044 TI - Microsatellite primer resource for Populus developed from the mapped sequence scaffolds of the Nisqually-1 genome. AB - In this study, 148 428 simple sequence repeat (SSR) primer pairs were designed from the unambiguously mapped sequence scaffolds of the Nisqually-1 genome. The physical position of the priming sites were identified along each of the 19 Populus chromosomes, and it was specified whether the priming sequences belong to intronic, intergenic, exonic or UTR regions. A subset of 150 SSR loci were amplified and a high amplification success rate (72%) was obtained in P. tremuloides, which belongs to a divergent subgenus of Populus relative to Nisqually-1. PCR reactions showed that the amplification success rate of exonic primer pairs was much higher than that of the intronic/intergenic primer pairs. Applying ANOVA and regression analyses to the flanking sequences of microsatellites, the repeat lengths, the GC contents of the repeats, the repeat motif numbers, the repeat motif length and the base composition of the repeat motif, it was determined that only the base composition of the repeat motif and the repeat motif length significantly affect the microsatellite variability in P. tremuloides samples. The SSR primer resource developed in this study provides a database for selecting highly transferable SSR markers with known physical position in the Populus genome and provides a comprehensive genetic tool to extend the genome sequence of Nisqually-1 to genetic studies in different Populus species. PMID- 19121048 TI - The role of race: should race be a factor in health care screening and treatment? PMID- 19121049 TI - The cost of being female: women and the individual insurance market. PMID- 19121050 TI - Screening for diabetes: managing illness before it occurs. PMID- 19121051 TI - Patient page. Diabetes: answers to common questions. PMID- 19121052 TI - Supporting consumers' access to research: how nurses can help patients use evidence to make medical decisions. PMID- 19121053 TI - Elective cesarean section: why women choose it and what nurses need to know. PMID- 19121054 TI - Breast and ovarian cancer: a new model for educating women. PMID- 19121055 TI - Type 2 diabetes: fueling the surge of cardiovascular disease in women. PMID- 19121056 TI - SBAR for maternal transports: going the extra mile. PMID- 19121057 TI - Simulation-based learning: how simulators help nurses improve clinical skills and preserve patient safety. PMID- 19121058 TI - Succession planning: why grooming their replacements is critical for nurse leaders. PMID- 19121059 TI - Preventing flu in health workers: vaccination challenge is launched. PMID- 19121061 TI - A most rewarding day. PMID- 19121062 TI - Evidence of the presence of T helper type 17 cells in chronic lesions of human periodontal disease. AB - INTRODUCTION: Periodontal disease is a chronic inflammation of the attachment structures of the teeth, triggered by potentially hazardous microorganisms and the consequent immune-inflammatory responses. In humans, the T helper type 17 (Th17) lineage, characterized by interleukin-17 (IL-17) production, develops under transforming growth factor-beta (TGF-beta), IL-1beta, and IL-6 signaling, while its pool is maintained by IL-23. Although this subset of cells has been implicated in various autoimmune, inflammatory, and bone-destructive conditions, the exact role of T lymphocytes in chronic periodontitis is still controversial. Therefore, in this study we investigated the presence of Th17 cells in human periodontal disease. METHODS: Gingival and alveolar bone samples from healthy patients and patients with chronic periodontitis were collected and used for the subsequent assays. The messenger RNA expression for the cytokines IL-17, TGF beta, IL-1beta, IL-6, and IL-23 in gingiva or IL-17 and receptor activator for nuclear factor-kappaB ligand in alveolar bone was evaluated by real-time polymerase chain reaction. The production of IL-17, TGF-beta, IL-1beta, IL-6, and IL-23 proteins was evaluated by immunohistochemistry and the presence of Th17 cells in the inflamed gingiva was confirmed by immunofluorescence confocal microscopy for CD4 and IL-17 colocalization. RESULTS: Our data demonstrated elevated levels of IL-17, TGF-beta, IL-1beta, IL-6, and IL-23 messenger RNA and protein in diseased tissues as well as the presence of Th17 cells in gingiva from patients with periodontitis. Moreover, IL-17 and the bone resorption factor RANKL were abundantly expressed in the alveolar bone of diseased patients, in contrast to low detection in controls. CONCLUSION: These results provided strong evidence for the presence of Th17 cells in the sites of chronic inflammation in human periodontal disease. PMID- 19121063 TI - Cytokines in gingival crevicular fluid of adolescents and young adults. AB - BACKGROUND/AIM: The purpose of this study was to compare the levels of the cytokines interleukin-1beta (IL-1beta), IL-4, and IL-8 in the gingival crevicular fluid (GCF) of adolescents and young adults. METHODS: Twenty-five adolescents aged between 14 and 16 years (Group A) and 20 periodontally healthy young adults aged between 25 and 35 years (Group B) were selected from two private dental clinics limited to pedodontics and periodontics respectively in Piraeus Greece. All subjects were systemically healthy. Clinical examination included probing pocket depth (PPD), presence or absence of plaque, and bleeding on probing (BOP). GCF was collected from four sites per subject. IL-1beta, IL-4, and IL-8, measured as total amounts (pg/30 s), were evaluated in 180 samples using a commercially available sandwich enzyme-linked immunosorbent assay. RESULTS: IL-1beta mean levels of Groups A and B were adjusted for BOP and PPD. Differences of IL-1beta mean levels between the two age groups were statistically significant (F = 50.245, P < 0.001) in favour of Group A. Adolescents showed statistically significantly lower mean levels of IL-4 than young adults in the presence of BOP (F = 10.690, P = 0.001). There was no statistically significant difference between adolescents and adults for the means of IL-8 adjusted for BOP and plaque presence (F = 2.032, P = 0.161). CONCLUSIONS: Within the limits of this study the differences reported in mean levels of IL-1beta and IL-4 may be attributed to the different age status. PMID- 19121064 TI - The host cytokine response to Porphyromonas gingivalis is modified by gingipains. AB - BACKGROUND/AIMS: Clinical studies indicate that primary proinflammatory cytokines, such as interleukin-1beta (IL-1beta) are elevated in the gingival crevice around teeth with periodontitis but the secondary cytokines and chemokines, IL-6 and IL-8, are not. The human gingival epithelial cells (HGECs) lining the gingival sulcus respond to perturbation by microbes of dental plaque by releasing a wide range of cytokines. Porphyromonas gingivalis, a putative periodontal pathogen, possesses numerous virulence factors some of which directly impact on the host response. In the present study, we sought to determine how P. gingivalis influences the inflammatory cytokine responses. METHODS: HGECs were challenged with P. gingivalis and other putative periodontal pathogens, and the resultant production of IL-1beta, IL-6, and IL-8 was assayed by enzyme-linked immunosorbent assay (ELISA). Culture supernatants and recombinant human cytokines were challenged with live P. gingivalis wild-type and gingipain-deficient strains and the resultant cytokine profile was assessed by ELISA and Western blot. RESULTS: We show here that primary HGECs challenged with live P. gingivalis result in high levels of IL-1beta but not the related secondary cytokines IL-6 and IL-8. We further demonstrate that cytokine response differences are the result of the action of P. gingivalis proteases, with lysine gingipain being the most effective. CONCLUSION: We conclude that P. gingivalis, through lysine gingipain, can subvert the protective host proinflammatory response by direct cytokine degradation. Changes in the crevicular cytokine profile have consequences in periodontal disease pathogenesis that should be considered in the development of diagnostic and therapeutic modalities. PMID- 19121066 TI - Factors involved in the T helper type 1 and type 2 cell commitment and osteoclast regulation in inflammatory apical diseases. AB - INTRODUCTION: Periapical chronic lesion formation involves activation of the immune response and alveolar bone resorption around the tooth apex. However, the overall roles of T helper type 1 (Th1), Th2, and T-regulatory cell (Treg) responses and osteoclast regulatory factors in periapical cysts and granulomas have not been fully determined. This study aimed to investigate whether different forms of apical periodontitis, namely cysts and granulomas, show different balances of Th1, Th2 regulators, Treg markers, and factors involved in osteoclast chemotaxis and activation. METHODS: Gene expression of these factors was assessed using quantitative real-time polymerase chain reaction, in samples obtained from healthy gingiva (n = 8), periapical granulomas (n = 20), and cysts (n = 10). RESULTS: Periapical cysts exhibited a greater expression of GATA-3, while a greater expression of T-bet, Foxp3, and interleukin-10 (IL-10) was seen in granulomas. The expression of interferon-gamma, IL-4, and transforming growth factor-beta was similar in both lesions. Regarding osteoclastic factors, while the expression of SDF-1alpha/CXCL12 and CCR1 was higher in cysts, the expression of RANKL was significantly higher in granulomas. Both lesions exhibited similar expression of CXCR4, CKbeta8/CCL23, and osteoprotegerin, which were significantly higher than in control. CONCLUSION: Our results showed a predominance of osteoclast activity in granulomas that was correlated with the Th1 response. The concomitant expression of Treg cell markers suggests a possible suppression of the Th1 response in granulomas. On the other hand, in cysts the Th2 activity is augmented. The mechanisms of periradicular lesion development are still not fully understood but the imbalance of immune and osteoclastic cell activity in cysts and granulomas seems to be critically regulated by Treg cells. PMID- 19121067 TI - Association between Bifidobacteriaceae and the clinical severity of root caries lesions. AB - BACKGROUND/AIMS: The isolation of members of the family Bifidobacteriaceae (bifids) from oral samples has been sporadic and a recent cloning study has suggested that they are not detectable in root caries lesions. METHODS: To better understand the presence of bifids in root caries we obtained clinical samples (15 of each) from sound exposed root surfaces, leathery remineralizing root lesions, and soft active root lesions. We investigated each for the presence of bifids using a mupirocin-containing selective medium and identified the isolates using 16S recombinant RNA sequencing. RESULTS: The proportion of bifids, as a percentage of the total anaerobic count, was significantly related to the clinical status of the sites sampled, being 7.88 +/- 1.93 in the infected dentine from soft lesions, 1.61 +/- 0.91 in leathery lesions, and 0.05 +/- 0.39 in plaque from sound exposed root surfaces. Bifids were isolated from all soft lesions, 13 of 15 leathery lesions, and five of the plaque samples. Bifidobacterium dentium was isolated from four of the plaque samples, from 13 samples from leathery lesions, and from 12 of the 15 samples of infected dentine from the soft active lesions. Parascardovia denticolens and Scardovia genomospecies C1 were each isolated from samples associated with all three clinical conditions whereas Scardovia inopicata and Bifidobacterium subtile were both isolated from the infected dentine of the leathery and soft lesions. Bifidobacterium breve was isolated from the infected dentine of soft root caries lesions. CONCLUSION: Bifids may be routinely isolated from root caries lesions using appropriate cultural methods. PMID- 19121065 TI - Multiple components contribute to ability of saliva to inhibit influenza viruses. AB - INTRODUCTION: Saliva is a potentially important barrier against respiratory viral infection but its mechanism of action is not well studied. METHODS: We tested the antiviral activities of whole saliva, specific salivary gland secretions, and purified salivary proteins against strains of influenza A virus (IAV) in vitro. RESULTS: Whole saliva or parotid or submandibular/sublingual secretions from healthy donors inhibited IAV based on hemagglutination inhibition and neutralization assays. This differs from human immunodeficiency virus (HIV), for which only submandibular/sublingual secretions are reported to be inhibitory. Among purified salivary proteins, MUC5B, scavenger receptor cysteine-rich glycoprotein 340 (salivary gp-340), histatins, and human neutrophil defensins (HNPs) inhibited IAV at the concentrations present in whole saliva. In contrast, some abundant salivary proteins (acidic proline-rich proteins and amylase) had no activity, nor did several other less abundant salivary proteins with known activity against HIV (e.g. thrombospondin or serum leukocyte protease inhibitor). Whole saliva and MUC5B did not inhibit neuraminidase activity of IAV and viral neutralizing and aggregating activity of MUC5B was potentiated by the neuraminidase inhibitor oseltamivir. Hence, MUC5B inhibits IAV by presenting a sialic acid ligand for the viral hemagglutinin. The mechanism of action of histatins requires further study. CONCLUSIONS: These findings indicate that saliva represents an important initial barrier to IAV infection and underline the complexity of host defense activity of oral secretions. Of interest, antiviral activity of saliva against IAV and HIV differs in terms of specific glandular secretions and proteins that are inhibitory. PMID- 19121068 TI - Toll-like receptors 2 and 5 in human gingival epithelial cells co-operate with T cell cytokine interleukin-17. AB - BACKGROUND/AIM: Periodontitis begins as the result of perturbation of the gingival epithelial cells caused by subgingival bacteria interacting with the epithelial cells via pattern recognition receptors. Toll-like receptors (TLRs) have been shown to play an important role in the recognition of periodontal pathogens so we have studied the interaction of TLR ligands with TLR2 and TLR5 for cytokine production in the cultures of gingival epithelial cells. METHODS: Immunohistochemistry was used for the localization of TLR2 and TLR5 in tissue specimens. Enzyme-linked immunosorbent assays were performed to detect the levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), released from gingival epithelial cell cultures following stimulation with TLR ligand alone or in combination with IL-17. RESULTS: Both TLR2 and TLR5 were increased in periodontitis (2128 +/- 159 vs. 449 +/- 59 and 2456 +/- 297 vs. 679 +/- 103, respectively, P < 0.001) including gingival epithelial cells that stained strongly. Cultured gingival epithelial cells stimulated with their respective ligands (HKLM, a TLR2 ligand that is also found in Porphyromonas gingivalis, and flagellin, a TLR5 ligand that is also found in Treponema denticola) produced both IL-1beta and TNF-alpha. To mimic T-cell help, IL-17 was added. This further greatly enhanced TLR ligand-induced IL-1beta (P < 0.001) and TNF-alpha (P < 0.01) production. CONCLUSIONS: These findings show how pathogen associated molecular patterns, shared by many different periodontopathogenic bacteria, stimulate the resident gingival epithelial cells to inflammatory responses in a TLR-dependent manner. This stimulation may be particularly strong in periodontitis and when T helper type 17 cells provide T-cell help in intercellular cooperation. PMID- 19121069 TI - Molecular analyses of bacterial DNA in extirpated heart valves from patients with infective endocarditis. AB - BACKGROUND/AIMS: Infective endocarditis (IE) is caused by a microbial infection of the endothelial surface of the heart. Although blood culture examinations are commonly used to determine the associated bacterial species, molecular techniques, which enable rapid identification of targeted bacterial species, have recently been applied in clinical cases. METHODS: Nine heart valve specimens from IE patients (six subacute cases and three acute cases) were extirpated and collected, then bacterial DNA was extracted. Bacterial species in the specimens were determined by two different molecular methods and the results were compared with those from a conventional blood culture technique. In addition, a comparison between the two molecular methods was carried out using known numbers of six streptococcal species. RESULTS: The conventional blood culture method revealed the bacterial species in eight cases, while one was found to be negative. Multiple species were identified in most of the cases by both molecular methods; however, those specified by one method were not always consistent with those specified by the other. Furthermore, the species determined by the blood culture technique were not always identified by the molecular methods. We also found that the two molecular methods used in the present study were extremely sensitive to detect from 1 to 100 cells of individual oral streptococcal species. CONCLUSION: Our results suggest that species specified by molecular methods may have disseminated incidentally into the bloodstream, so interpretation of such results should be carefully undertaken in clinical situations. PMID- 19121070 TI - Nitric oxide production by a human osteoblast cell line stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide. AB - BACKGROUND/AIM: Human osteoblasts induced by inflammatory stimuli express an inducible nitric oxide synthase (iNOS). The aim of the present study was to test the hypothesis that Aggregatibacter actinomycetemcomitans lipopolysaccharide stimulates the production of nitric oxide (NO) by a human osteoblast-like cell line (HOS cells). METHODS: Cells were stimulated directly with A. actinomycetemcomitans lipopolysaccharide or pretreated with the following l-NIL (an iNOS inhibitor), anti-CD14, Toll-like receptor 2 (TLR2), or TLR4 antibody before stimulation with A. actinomycetemcomitans lipopolysaccharide. The role of the cyclic nucleotides was assessed by pretreating the cells with the following; ODQ (a guanylyl cyclase inhibitor); SQ22536 (an adenylyl cyclase inhibitor); db cAMP (a cyclic adenosine monophosphate analog); br-cGMP (a cyclic guanosine monophosphate analog); forskolin (an adenylyl cyclase activator), IBMX [a non specific phosphodiesterase (PDE) inhibitor], or KT5720 [a protein kinase A (PKA) inhibitor]. The cells were also preincubated with genistein [a protein tyrosine kinase (PTK) inhibitor], bisindolylmaleimide [a protein kinase C (PKC) inhibitor], BPB [a phospholipase A2 (PLA2) inhibitor], and NDGA (a lipoxygenase inhibitor). The iNOS activity and nitrite production in the cell cultures were determined spectrophotometrically. RESULTS: The results showed that A. actinomycetemcomitans lipopolysaccharide stimulated both iNOS activity and nitrite production by HOS cells; this was reduced by l-NIL, anti-CD14, or anti TLR4 antibody, SQ22536, KT5720, genistein, bisindolylmaleimde, BPB, and NDGA, but was enhanced by db-cAMP, IBMX, and forskolin. CONCLUSION: These results therefore suggest that A. actinomycetemcomitans lipopolysaccharide may induce the production of NO by HOS cells via a CD14-TLR4 molecule complex, a cAMP-PKA pathway, as well as by a PTK, PKC, PLA2, and lipoxygenase-dependent mechanism. PMID- 19121071 TI - Zinc and copper play a role in coaggregation inhibiting action of Porphyromonas gingivalis. AB - BACKGROUND/AIM: We investigated the mechanisms of adherence of salivary and serum proteins, which mimic gingival crevicular fluid (GCF), to Porphyromonas gingivalis, and the effects of these adhered proteins on coaggregation and hemagglutination properties. METHODS: The amounts of salivary and serum proteins adhering to P. gingivalis were determined using (3)H-labeled and non-labeled proteins. The coaggregation between P. gingivalis and Streptococcus oralis or Streptococcus gordonii was observed. Hemagglutination was evaluated using sheep erythrocytes. Proteins that interacted with zinc or copper in saliva and serum and on P. gingivalis were examined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: The amount of salivary or serum proteins that adhered to the surface of P. gingivalis strains was increased by cations, especially zinc and copper ions. The pretreatment of bacterial cells with salivary or serum proteins before the assay inhibited coaggregation with gram positive bacteria and hemagglutination. These phenomena were enhanced by the presence of zinc or copper ions during the pretreatment of P. gingivalis with proteins. We detected protein bands that were related to these cations in saliva and serum and on P. gingivalis. CONCLUSIONS: Our findings suggest that zinc and copper ions markedly enhanced the adhesion and accumulation of salivary and serum proteins on cells of P. gingivalis and inhibited the coaggregation and hemagglutination of P. gingivalis. These cations might be useful for limiting the settlement of P. gingivalis in the gingival sulcus with the goal of preventing periodontal disease. PMID- 19121072 TI - Detection of oral bacteria in cardiovascular specimens. AB - BACKGROUND/AIMS: Oral bacteria, including cariogenic and periodontal pathogens, are thought to be etiological factors in the development of cardiovascular diseases. To define this relationship, we analyzed the distribution of oral bacterial species in cardiovascular specimens. METHOD: Following acceptance into the study, 203 consecutive patients were analyzed, from whom 82 aortic valve specimens, 35 mitral valve specimens, and 86 aortic aneurysmal wall specimens, of which 16 contained aneurysmal thrombus tissues, were obtained. In addition, a total of 58 dental plaque specimens were collected from the same group of patients who underwent heart valve replacement or removal of aortic aneurysms. Bacterial DNA was extracted from both cardiovascular tissues and dental plaque in those cases and then species-specific polymerase chain reaction assays were used to analyze the occurrences of six oral streptococcal and six periodontal bacterial species. RESULTS: Streptococcus mutans was the most frequently detected species in the cardiovascular specimens, followed by Aggregatibacter actinomycetemcomitans. As for dental plaque specimens from patients who underwent cardiovascular operations, most of the tested periodontitis-related species as well as oral streptococci were detected at high frequencies. Furthermore, the positive rate of S. mutans in cardiovascular specimens from patients whose dental plaque specimens were also positive for S. mutans was 78%, which was significantly higher than any other tested species when the same analysis was performed. CONCLUSION: Our results suggest that specific oral bacterial species, such as S. mutans and A. actinomycetemcomitans, are related to bacteremia and may be etiologic factors for the development of cardiovascular diseases. PMID- 19121073 TI - Application of stereological principles for quantification of bacteria in intact dental biofilms. AB - INTRODUCTION: Quantative confocal laser scanning microscopy (CLSM) in combination with fluorescent in situ hybridization (FISH) may help to increase our knowledge about biofilm formation. The purpose of this study was to develop and evaluate a stereological method for quantification of bacteria in intact biofilm. The method was applied in a quantitative study of the proportion of streptococci relative to other bacteria in initial in-situ-grown dental biofilms as a function of time. METHODS: Biofilms were collected on standardized glass slabs mounted in intra oral appliances and worn by 10 individuals for 6, 12, 24, and 48 h. Biofilms were analysed using CLSM. Quantification of bacteria labelled with 16S ribosomal RNA oligonucleotide probes was performed with stereological tools: the unbiased counting frame and the two-dimensional fractionator. RESULTS: Results showed a notable increase in the total number of bacteria and streptococci over time, with a considerable inter-individual variation at each time-point. After 48 h there was a 12.5-fold difference between individuals in the total number of bacteria and a 12.6-fold difference in the number of streptococci. The number of streptococci exceeded that of other bacteria and over the examination period there was a relatively constant relationship between the number of streptococci and other bacteria (streptococci vs. non-streptococci: median 15.2; minimum 1.0; maximum 89.3). CONCLUSION: The study demonstrates that the combined use of FISH and stereology is a relevant and reliable tool for obtaining unbiased information about the numerical contributions of specific bacterial populations during early biofilm formation. PMID- 19121074 TI - Oral candidiasis: a comparison between conventional methods and multiplex polymerase chain reaction for species identification. AB - BACKGROUND/AIM: Oral candidiasis is the most common fungal infection in dental practice, and is caused by yeasts that are normally present in the endogenous flora. METHODS: To evaluate a rapid diagnostic method for identification of Candida oral isolates, a multiplex polymerase chain reaction (PCR) was carried out on colonies and on oral rinse solutions from 95 subjects with suspected oral candidiasis and results were compared with those from seven commonly used phenotypic identification systems. RESULTS: Between four and nine species were characterized in the samples by the phenotypic methods. PCR identified the same species in 60 (74%) samples from both colony and oral rinse solutions. Statistical analysis, carried out only for the three most frequently isolated species (Candida albicans, Candida glabrata, and Candida tropicalis), showed good concordance in the comparison of multiplex PCR with API 20C AUX and with the Rapid Yeast Identification Panel; conversely, significant differences were registered in the comparison between the molecular method and other phenotypic systems, including four chromogenic media and the automated system Vitek2. DISCUSSION: Multiplex PCR was rapid and effective in the identification of Candida species and allowed the detection of more than one species in the same sample. PMID- 19121075 TI - Distribution, regulation and role of the agmatine deiminase system in mutans streptococci. AB - The agmatine deiminase system (AgDS) was identified in seven strains of mutans streptococci. Genes encoding the AgDS of Streptococcus rattus FA-1 were sequenced and found to share homology with the agu genes of Streptococcus mutans UA159. With the exception of Streptococcus sobrinus, the AgDS of mutans streptococci appear to be sensitive to carbohydrate catabolite repression. Agmatine inhibited bacterial growth, suggesting that the AgDS degrades a deleterious substance into useful compounds. PMID- 19121076 TI - Gene targeting demonstrates that inducible nitric oxide synthase is not essential for resistance to oral candidiasis in mice, or for killing of Candida albicans by macrophages in vitro. AB - INTRODUCTION: Oral candidiasis is caused by opportunistic infections with the yeast Candida albicans. Previous studies have demonstrated important roles for innate immunity and T helper type 1-mediated inflammatory reactions in recovery from infection, with macrophages and neutrophils as key effector cells. Both effector cell types use the inducible isoform of nitric oxide synthase (iNOS) to generate candidacidal molecules, but it is not clear whether nitric oxide (NO) is an absolute requirement for candidacidal effector activity. METHODS: In this study we directly investigated the role of iNOS-derived NO in resistance to murine experimental oral candidiasis, using iNOS knockout mice. RESULTS: Knockout mice were no more susceptible to oral candidiasis than wild-type controls. Bone marrow-derived macrophages from the knockout mice killed C. albicans yeasts efficiently in vitro, and were still able to produce nitrites in an iNOS independent manner, albeit less efficiently than wild-type controls. There were no significant differences in local mucosal production of interleukins 6, 12, 17A, or 23, interferon-gamma, or transforming growth factor-beta 24 h after oral challenge with C. albicans. CONCLUSION: These data suggest that iNOS-derived NO is not required for resistance to oral candidiasis in vivo, and that bone marrow derived macrophages may have iNOS-independent means of generating reactive nitrogen species. PMID- 19121077 TI - Parasite Immunology has established itself over the last 30 years as one of the leading specialist journals in the field. Editorial. PMID- 19121078 TI - Interferon-gamma-mediated activation of enterocytes in immunological control of Encephalitozoon intestinalis infection. AB - The microsporidian Encephalitozoon intestinalis develops within intestinal epithelial cells (enterocytes) and is an important opportunistic diarrhoeal pathogen associated with AIDS. Little is known about the protective immune response against the parasite although in mice IFN-gamma is involved and is required to prevent dissemination of the infection to other organs. The present study was designed to establish a suitable short-term in vitro culture technique for E. intestinalis that would enable studies of the role of cytokines such as IFN-gamma in the effector phase of immunity. Encephalitozoon intestinalis reproduced considerably better in the murine enterocyte cell line CMT-93 than in the three human enterocyte cell lines Caco-2, HT29 and HCT-8. Treatment of CMT-93 cells with IFN-gamma significantly reduced parasite reproduction in a dose- and time-dependent manner. IFN-gamma also inhibited development of the parasite in Caco-2 cells. Neither production of NO nor Fe deprivation appeared to be involved in IFN-gamma-mediated parasite killing. However studies suggested that tryptophan catabolism by indoleamine 2,3-dioxygenase played an important part in inactivation of E. intestinalis. PMID- 19121079 TI - Proteomic analysis of excretory/secretory products released by Teladorsagia circumcincta larvae early post-infection. AB - Teladorsagia circumcincta is an important parasitic nematode of domestic small ruminants. Drug resistance in this species is common so alternative methods of control are required. As animals develop immunity to T. circumcincta, vaccination is a valid option. Little is known about the antigens that play a role in stimulating immunity at this host/parasite interface. As responses generated between 1 and 5 dpi are known to affect development of these nematodes in their gastric niche, we focused on proteins released during the early stages of infection. To identify molecules potentially involved in immunity, we undertook a proteomics analysis of proteins released from larvae harvested at 1-, 3- and 5 days post-infection (dpi). This analysis produced peptide sequence data that was used to search information available in T. circumcincta expressed sequence tag (EST) databases and enabled identification of a number of excretory/secretory (ES) proteins. Immunoblots were performed to assess the relative molecular weight of ES antigens that were targets of local IgA responses in mucus from sheep rendered immune to infection. ELISA was performed to assess antigen-specific mucus IgA levels in individual sheep. These experiments provided preliminary evidence that the proteins identified in the larval secretome were subject to these antibody responses. PMID- 19121080 TI - Germinal centre and marginal zone B cells expand quickly in a second Plasmodium chabaudi malaria infection producing mature plasma cells. AB - Antibodies and B cells are critical in the protective immune response to the blood stage of the malaria parasite, Plasmodium chabaudi. However, little is known about the development of memory B cells and their differentiation into plasma cells during infection or after re-infection. Here we have shown that B cells with phenotypic characteristics of memory cells (CD19(+)IgD(-) CD38(+), IgG1(+)) are generated in a primary Plasmodium chabaudi chabaudi infection of mice. In addition, we observed that germinal centre cells (CD19(+), GL7(+), MHCII(hi)) and Marginal Zone B cells (CD19(+)CD23(-)IgD(-)) show faster expansion on re-infection than in the primary, though other subsets do not. Interestingly, though both IgM(-) and IgM(+) memory cells are produced, IgM(+) memory cells do not expand on second infection. The second infection quickly produced mature bone marrow plasma cells (intracellular Ig(hi), CD138(hi), CD9(+), B220(-)), compared to primary infection; which generates a very large population of immature splenic plasma cells (B220+). This analysis suggests that a memory B cell population is generated after a single infection of malaria, which on re-infection responds quickly producing germinal centres and generating long-lived plasma cells making the second encounter with parasite more efficient. PMID- 19121081 TI - Immune recognition of the surface associated antigen, Tc-SAA-1, from infective larvae of Teladorsagia circumcincta. AB - A cDNA encoding a surface-associated antigen was amplified by reverse transcriptase polymerase chain reaction (PCR) from RNA extracted from Teladorsagia circumcincta exsheathed third stage larvae (xL3). The protein encoded by this cDNA, Tc-SAA-1, displays 77% identity over 162 amino acid residues to a surface associated antigen from Ancylostoma caninum (Ac-SAA-1). Antiserum raised against a bacterially-expressed recombinant form of Tc-SAA-1 reacted with a native protein in somatic and surface extracts of xL3 but not with L4 or adult parasites. Limited binding of anti-Tc-SAA-1 antibody was observed on the cuticular surface of xL3 s, however, regions of localization underlying the cuticle were observed. Incubation of xL3 T. circumcincta with anti-SAA rabbit serum failed to significantly inhibit penetration of the abomasal mucosa in vitro. IgA in abomasal mucus derived from sheep that had received a trickle infection of T. circumcincta bound recombinant Tc-SAA-1. PMID- 19121082 TI - Protein-energy malnutrition decreases immune response to Leishmania chagasi vaccine in BALB/c mice. AB - Protein-energy malnutrition and visceral leishmaniasis are important problems of public health affecting millions of people worldwide. Vaccine efficacy depends on the ability of individuals to mount an appropriate immune response and may be inadequate in malnourished persons. In this study, we used a mouse model to verify the effect of combined protein, iron and zinc deficiency in the response to Leishmania chagasi antigen vaccine. BALB/c mice were fed with a low-protein (3% casein), iron- and zinc-deficient diet or control diet (14% casein and sufficient in zinc and iron). After malnutrition establishment, mice were vaccinated subcutaneously with L. chagasi Ag plus saponin. After vaccination, mice were nutritionally repleted and then all mice were challenged with L. chagasi promastigotes. Four weeks later, liver and spleen parasite load was evaluated. Our data show that vaccine caused a significant reduction in parasite load in spleen and liver from mice fed with control diet. However, splenic parasitism was increased in mice fed with deficient diet and this diet caused a reduction in splenocyte IFN-gamma production in response to the vaccine in repleted mice. These data suggest that malnutrition may alter immune response to L. chagasi vaccine in BALB/c model of infection, even after nutritional repletion. PMID- 19121083 TI - Effect of Nippostrongylus brasiliensis L3 ES on inflammatory mediator gene transcription in lipopolysaccharide lung inflammation. AB - The anti-inflammatory properties of parasitic helminths have been largely linked to their excretory-secretory (ES) products. Some studies have noted a lack of TNF alpha production and limited recruitment of neutrophils into the lungs after Nippostrongylus brasiliensis infection. We previously reported that instillation of ES from L3 larvae of N. brasiliensis to the lungs could inhibit the recruitment of neutrophils on a background of LPS-induced inflammation. A similar reduction in neutrophil recruitment was observed in this study. This reduction was associated with the significant inhibition in gene transcription of the adhesion molecule, ICAM-1, and the chemokine, MIP-2 in bronchoalveolar lavage (BAL) cells. The LPS-stimulated gene transcription of the pro-inflammatory cytokines TNF-alpha and IL-1beta was also significantly reduced by L3 ES. Inducible nitric oxide synthase (iNOS) is normally elevated in classically activated macrophages, however, in this case gene transcription of iNOS was inhibited by L3 ES and may suggest a phenotype change to anti-inflammatory. The general inhibition of pro-inflammatory mediators observed in this study suggests that infective stage L3 larvae excrete and/or secrete inhibitory products capable of modifying the normally potent LPS inflammatory response. PMID- 19121086 TI - Clinicopathology of stromal invasion in lung adenocarcinoma. AB - In the World Health Organization classification, lung adenocarcinoma with mixed subtypes is defined as invasive carcinoma with evidence of vascular, pleural, or stromal invasion. The histological criteria for stromal invasion, however, are not clearly established. A total of 157 peripheral pure bronchioloalveolar carcinoma (BAC) or lung adenocarcinoma with mixed BAC and others were reviewed. All cases had been resected between 1986 and 2000 and measured < or =30 mm in maximum dimension. Destruction of alveolar framework (DAF) was defined as distortion or discontinuity of the alveolar framework by tumor growth. The extra alveolar area involvement (EAAI) was defined as tumor growth outside the alveolar framework, which includes the following areas: bronchial wall, perivascular connective tissue and/or the vascular wall, interlobular septum and the visceral pleura. Survival of patients with adenocarcinoma without DAF (n = 41) was 100%. Even when adenocarcinoma involved DAF and lacked EAAI (n = 21), survival was 100%. The 5 year survival rate of groups with two invasion signs (n = 34) was 90.1%, and that of groups with three to five invasion signs (n = 61) was 66.7%. Tumor growth outside the alveolar framework is the hallmark of stromal invasion. PMID- 19121087 TI - L-type amino-acid transporter 1 as a novel biomarker for high-grade malignancy in prostate cancer. AB - To find reliable biomarkers for high-grade malignancy, the relationship between immunohistochemical L-type amino-acid transporter 1 (LAT1) expression of biopsy samples, determined with the newly developed monoclonal antibody against human LAT1, and prognosis of patients with prostate cancer, was investigated. The intensity and score of immunohistochemical LAT1 expression of first biopsy samples were assessed using the modified Sinicrope et al. method and were found to be correlated with poor survival for the study group of 114 surgically treated patients as a whole (P = 0.0002 and 0.0270, respectively). LAT1 intensity further had a significant relationship (P = 0.0057) with prognosis in pathological T3 + T4 groups. Multivariate analysis indicated that the LAT1 intensity and score were more reliable prognostic markers, compared with the Gleason score and the Ki-67 labeling index. A relationship of the LAT1 intensity and score with prognosis could also be confirmed in 63 patients with inoperable cancer (P = 0.0070 and <0.0001, respectively). Similarly, significant differences in prognosis were confirmed in clinical T3 + T4 groups (P = 0.0091 and 0.0244, respectively). Moreover, the combination of LAT1 expression and Gleason score was found to have a more reliable correlation with prognosis. Thus, elevated LAT1 expression in prostate cancers is a novel independent biomarker of high-grade malignancy, which can be utilized together with the Gleason score, which is mainly dependent on cellular and structural atypia, to assess prognosis. PMID- 19121088 TI - Therapeutic strategy targeting the mTOR-HIF-1alpha-VEGF pathway in ovarian clear cell adenocarcinoma. AB - Malignant tumors usually involve a relatively hypoxic state, which induces overexpression of hypoxia-inducible factor-1alpha (HIF-1alpha) to satisfactorily enable the tumor to survive. Thus, inhibition of the mammalian target of rapamycin (mTOR) pathway including HIF-1alpha is expected to play a major role in suppression of tumor cell growth, having recently drawn much attention as an anti cancer therapeutic strategy for various malignant tumors. In the present study, which compared clear cell adenocarcinoma (CLA) of the ovary with serous adenocarcinoma (SEA), the immunohistochemical expression of mTOR, phosphorylated mTOR (p-mTOR), HIF-1alpha, and vascular endothelial growth factor (VEGF) was examined in surgically resected specimens of 29 SEA and 47 CLA. There were no significant differences in expression of mTOR, HIF-1alpha and VEGF between SEA and CLA, but it was noted that p-mTOR expression was more prominent in CLA than SEA. Then, using the cell lines of CLA (RMG-1 and W3uF), an experimental study was designed to clarify whether tumor suppression due to downregulation of mTOR activity could represent a promising therapeutic strategy for CLA. After treatment of an analogue of rapamycin (everolimus), expression of mTOR, p-mTOR, HIF-1alpha and VEGF was examined on western blot. As a result, although mTOR expression remained unchangeable, expression of p-mTOR, HIF-1alpha and VEGF was shown to be sharply depressed. The same expression alterations were demonstrated in the xenograft model treated with everolimus. In conclusion, mTOR-targeted therapy through usage of drugs such as everolimus may be more effective for CLA of the ovary because of its significant expression of p-mTOR. PMID- 19121089 TI - Role of 3'-phosphoinositides in oncogenic KRAS-induced modulation of shape and motility of airway epithelial cells. AB - The authors' previous study demonstrated that oncogenic KRAS modulates the shape and motility of airway epithelial cells. To explore detailed mechanism mediating these events, the possible involvement of phosphatidylinositides (PIP) was investigated. The intracellular localization of PIP was visualized with a pleckstrin homology domain-enhanced green fluorescent protein (EGFP) construct. PIP accumulated at the leading edges of polarizing epithelial cells, while they co-localized with cortical actin at cell-cell contacts, suggesting that PIP play important roles in the cytoskeletal organization. Transduction of oncogenic KRAS induced multiple pseudopodia and disrupted cortical actin, enhancing motility. A mitogen activated protein kinase kinase (MEK) inhibitor reduced the accumulation of PIP at membranes and development of pseudopodia, and restored stable cortical actin, reducing the motility. A phosphoinositide 3-kinase (PI3K) inhibitor also reduced accumulation of PIP at membranes, formation of pseudopodia and motility, but its effect on cortical actin was indistinct. The KRAS V12/S35 mutant, activating only the MEK pathway, induced multiple pseudopodia and disrupted the cortical actin. The KRAS V12/C40 mutant, activating only the PI3K pathway, also induced pseudopodia, but its effect on cortical actin was obscure. Taken together, oncogenic KRAS could cause the accumulation of PIP via the PI3K and MEK pathways and modulate the cell shape and migration. PMID- 19121090 TI - Epithelioid angiomyolipoma of the kidney. AB - Epithelioid angiomyolipoma (eAMLoma) is an uncommon renal mesenchymal tumor with malignant potential and is frequently associated with tuberous sclerosis (TSC). It is composed of polygonal large-sized tumor cells arranged in an epithelioid manner. Differential diagnosis from renal cell carcinoma (RCC) is often challenging because of its epithelioid morphology. Herein is reported three cases of eAMLoma, involving one in a 28-year-old man with TSC and two in women without TSC (34 and 62 years of age, respectively). The male TSC patient had microscopic conventional AMLomas in the same kidney. All patients were positive for melanoma (reactive with HMB45 antibody, and positive for melan A, tyrosinase and microphthalmia transcription factor) and smooth muscle markers (positive for alpha-smooth muscle-specific actin), but not for epithelial markers (cytokeratin, epithelial membrane antigen). In particular, the translocation RCC is an important differential diagnostic candidate, in terms of the positive reaction with HMB45 and morphological similarity. The present tumor samples did not show any reactivity for transcription factor binding to IGHM enhancer 3 or transcription factor EB, which excluded the possibility of translocation RCC. The possibility of eAMLoma should be evaluated as a diagnostic candidate, especially in cases of renal tumors (i) in young patients; (ii) associated with TSC; or (iii) with an epithelioid morphology and a high nuclear grade. PMID- 19121091 TI - Adult teratoid Wilms' tumor with prominent neuroepithelial differentiation. AB - Teratoid Wilms' tumor is a rare variant of Wilms' tumor (WT) that has been reported exclusively in pediatric patients. The present paper describes a teratoid WT in a 50-year-old Korean man with a giant right renal mass. Radical nephrectomy was performed under the impression that the mass was a renal cell carcinoma. Grossly, the removed kidney contained a giant well-encapsulated mass measuring 24 x 18 x 10 cm with cystic changes, necrosis, and hemorrhage. On microscopy triphasic patterns of WT and prominent heterologous components were seen, including rhabdomyoblasts, neuroepithelial tissue, fat, cartilage, and various types of mature epithelium. This is the first reported case of a teratoid WT occurring in an adult kidney. PMID- 19121092 TI - Multifocal metanephric adenoma in childhood. AB - Metanephric adenoma is the most commonly occurring member of the metanephric tumor family, which also includes metanephric adenofibroma and metanephric stromal tumor. According to the World Health Organization classification, however, it is not commonly multifocal. Reported herein is the case of a 9-year old boy with multifocal metanephric adenoma. Histologically, surgical sections showed multifocal proliferation of small rounded and uniform cells with smooth nuclear contours, scant pale-staining cytoplasm, dark-staining nuclei, and inconspicuous nucleoli: the cells were arranged in sheets and acinal, ductal, glomeruloid, and papillary structures. On immunohistochemistry the tumor cells were positive for vimentin, cytokeratins (CAM5.2, AE1/AE3, and CK18), and WT1, but negative for cytokeratin 7 (CK7) and epithelial membrane antigen (EMA). The Ki-67 labeling index was <1%. In addition, cytogenetic analysis indicated a normal karyotype (46XY). Other histologically similar tumors are papillary renal cell carcinoma and nephroblastoma, and it is necessary to distinguish metanephric adenoma from those tumors because of malignancy. In contrast to those tumors, metanephric adenoma has inconspicuous nucleoli, loss of CK7 and EMA expression, and no mitotic figures. Thus, the histological and immunohistochemical features of the present case were compatible with metanephric adenoma. PMID- 19121093 TI - Intraductal tubular carcinoma, intestinal type, of the pancreas. AB - Presented herein is an unusual case of intraductal tubular carcinoma, intestinal type, of the pancreas. This tumor was characterized by intraductal adenoma with a few malignant foci, and also by entire involvement of the main pancreatic duct and no involvement of its branches. A 67-year-old man was admitted to hospital because of abdominal pain. On endoscopy and endoscopic retrograde cholangiopancreatography, irregular pancreatic duct was seen. No mucus secretion was observed on endoscopy. Because a biopsy showed tubular atypical cells, pancreato-duodenectomy was performed. Grossly, the entire main pancreatic duct had intraductal tumor, sparing its branches. No intraductal mucus was noted. Microscopically, the entire main pancreatic duct had proliferation of tubular adenomatous tumor without secretory mucins. Goblet cells were present in some areas. No pyloric type tubules were recognized. Malignant transformation was present in a few areas. No invasive features were recognized. On mucin histochemistry the tumor cell cytoplasm contained a little or no neutral and acidic mucus, and no secretory mucins were recognized. Immunohistochemically, the tumor cells were positive for cytokeratins (CK), CK 8, 9, 18, 19 and 20, epithelial membrane antigen, CDX2, carbohydrate antigen 19-9, and Ki-67 (labeling 30%), MUC2, MUC5AC and MUC6, and CD10. The tumor cells were negative for C-erbB2, MUC1, trypsin, pancreatic amylase and pancreatic lipase. The tumor cells were negative for p53 protein, but the malignant foci were positive for p53 protein and had high Ki-67 antigen (labeling 60%). The patient was free of disease 4 years after the operation. In summary, presented here is an extremely rare case of intraductal tubular carcinoma, intestinal type, showing focal malignant foci. PMID- 19121094 TI - Immunoreactivity of a monoclonal antibody against human placental alkaline phosphatase. PMID- 19121095 TI - Detecting hydrogen peroxide in leaves in vivo - a comparison of methods. AB - Four hydrogen peroxide detecting probes, 3,3'-diaminobenzidine (DAB), Amplex Red (AR), Amplex Ultra Red (AUR) and a europium-tetracycline complex (Eu(3)Tc) were infiltrated into tobacco leaves and tested for sensitivity to light, toxicity, subcellular localization and capacity to detect H(2)O(2) in vivo. In the absence of leaves, in water solutions, AUR was very much sensitive to strong light, AR showed slight light sensitivity, while DAB and Eu(3)Tc were insensitive to irradiation. When infiltrated into the leaves, the probes decreased the photochemical yield (Phi(PSII)) in the following order of effect AR > DAB > AUR > Eu(3)Tc. With the exception of Eu(3)Tc, all probes stimulated the build-up of non photochemical quenching either temporally (DAB, AUR) or permanently (AR), showing that their presence may already limit the photosynthetic capacity of leaves, even in the absence of additional stress. This should be taken into account when using these probes in plant stress experiments. Confocal laser scanning microscopy studies with the three fluorescent H(2)O(2) probes showed that the localizations of Eu(3)Tc and AUR were mainly intercellular. AR partly penetrated into leaf chloroplasts but probably not into the thylakoid membranes. Photosynthesis related stress applications of AR seem to be limited by the low availability of internal leaf peroxidases. Applications of AR for kinetic H(2)O(2) measurements would require a co-infiltration of external peroxidase, imposing another artificial modifying factor and thus taking experiments further from ideal, in vivo conditions. Our results suggest that the studied H(2)O(2) probes should be used in leaf studies with caution, carefully balancing benefits and artifacts. PMID- 19121096 TI - Ureide metabolism during seedling development in French bean (Phaseolus vulgaris). AB - French bean (Phaseolus vulgaris) is a legume that transports most of the atmospheric nitrogen fixed in its nodules to the aerial parts of the plant as ureides. Changes in ureide content and in enzymatic activities involved in their metabolism were identified in the cotyledons and embryonic axes during germination and early seedling development. Accumulation of ureides (ca. 1300 nmol per pair of cotyledons) was observed in the cotyledons of dry seeds. Throughout germination, the total amount of ureides slightly decreased to about 1200 nmol, but increased both in cotyledons and in embryonic axes after radicle emergence. In the axes, the ureides were almost equally distributed in roots, hypocotyls and epicotyls. The pattern of ureide distribution was not affected by the presence of nitrate or sucrose in the media up to 6 days after imbibition. Ureides are synthesized from purines because allopurinol (a xanthine dehydrogenase inhibitor) blocks the increase of ureides. Allantoin and allantoate degrading activities were detected in French bean dried seeds, whereas no ureidoglycolate-degrading activity was detected. During germination, the levels of the three activities remain unchanged in cotyledons. After radicle emergence, the levels of activities in cotyledons changed. Allantoin-degrading activity increased, allantoate-degrading activity decreased and ureidoglycolate-degrading activity remained undetectable in cotyledons. In developing embryonic axes, the three activities were detected throughout germination and early seedling development. The embryonic axes are able to synthesize ureides, because those compounds accumulated in axes without cotyledons. PMID- 19121097 TI - The oxidative stress caused by salinity in two barley cultivars is mitigated by elevated CO2. AB - Changes in antioxidant metabolism because of the effect of salinity stress (0, 80, 160 or 240 mM NaCl) on protective enzyme activities under ambient (350 micromol mol(-1)) and elevated (700 micromol mol(-1)) CO(2) concentrations were investigated in two barley cultivars (Hordeum vulgare L., cvs Alpha and Iranis). Electrolyte leakage, peroxidation, antioxidant enzyme activities [superoxide dismutase (SOD), EC 1.15.1.1; ascorbate peroxidase (APX), EC 1.11.1.11; catalase (CAT), EC 1.11.1.6; dehydroascorbate reductase (DHAR), EC 1.8.5.1; monodehydroascorbate reductase (MDHAR), EC 1.6.5.4; glutathione reductase (GR), EC 1.6.4.2] and their isoenzymatic profiles were determined. Under salinity and ambient CO(2), upregulation of antioxidant enzymes such as SOD, APX, CAT, DHAR and GR occurred. However, this upregulation was not enough to counteract all ROS formation as both ion leakage and lipid peroxidation came into play. The higher constitutive SOD and CAT activities together with a higher contribution of Cu,Zn SOD 1 detected in Iranis might possibly contribute and make this cultivar more salt-tolerant than Alpha. Elevated CO(2) alone had no effect on the constitutive levels of antioxidant enzymes in Iranis, whereas in Alpha it induced an increase in SOD, CAT and MDHAR together with a decrease of DHAR and GR. Under combined conditions of elevated CO(2) and salinity the oxidative damage recorded was lower, above all in Alpha, together with a lower upregulation of the antioxidant system. So it can be concluded that elevated CO(2) mitigates the oxidative stress caused by salinity, involving lower ROS generation and a better maintenance of redox homeostasis as a consequence of higher assimilation rates and lower photorespiration, being the response dependent on the cultivar analysed. PMID- 19121098 TI - Chloroplastic glutamine synthetase is activated by direct binding of aluminium. AB - Acidification of soils may release water soluble, toxic aluminium species from clay minerals. Al interferes with a wide range of physical and cellular processes. Glutamine synthetase (GS, EC 6.3.1.2) is the key enzyme of primary N assimilation, as well as ammonia reassimilation and detoxification. Plant GS requires two magnesium ions per subunit for activity, which makes GS a potential target of metal stress. The objective of this investigation was to prove that Al from an organic metal complex is able to activate GS, and Al becomes bound to the polypeptide structure of the GS molecule. Aluminium(III)-nitrilotriacetic acid complex (Al(III)NTA) activated the GS prepared from wheat (Triticum aestivum L.) leaves, as Al(3+) did in vivo, but could not functionally substitute magnesium ions, which were also necessary for the activity in the in vitro GS assay. GS2 was isolated by non-denaturing polyacrylamide gel electrophoresis, and the Al and Mg content of the enzyme was determined by inductively coupled plasma atomic emission spectroscopy. The GS octamer remained intact and contained Mg(2+) bound to its specific sites after the electrophoretic separation. Al was detected in the Al(III)NTA-treated sample bound to the structure of the enzyme protein, potentially occupying one of the specific metal-binding sites of the subunits. Our results indicate that the activatory effect of the Al(III)NTA complex is because of specific binding of aluminium to the polypeptide chain of GS2, however presence of magnesium at least on one of the metal-binding sites is essential to the active state of the enzyme. PMID- 19121099 TI - Effects of NaCl on responses of ectomycorrhizal black spruce (Picea mariana), white spruce (Picea glauca) and jack pine (Pinus banksiana) to fluoride. AB - Black spruce (Picea mariana), white spruce (Picea glauca) and jack pine (Pinus banksiana) were inoculated with Suillus tomentosus and subjected to potassium fluoride (1 mM KF and 5 mM KF) in the presence and absence of 60 mM NaCl. The NaCl and KF treatments reduced total dry weights in jack pine and black spruce seedlings, but they did not affect total dry weights in white spruce seedlings. The addition of 60 mM NaCl to KF treatment solutions alleviated fluoride-induced needle injury in ectomycorrhizal (ECM) black spruce and white spruce, but had little effect in jack pine seedlings. Both KF and 60 mM NaCl treatments reduced E values compared with non-treated control seedlings. However, with the exception of small reductions of K(r) by NaCl treatments in black spruce, the applied KF and NaCl treatments had little effect on K(r) in ECM plants. Chloride tissue concentrations in NaCl-treated plants were not affected by the presence of KF in treatment solutions. However, shoot F concentrations in ECM black spruce and white spruce treated with 5 mM KF + 60 mM NaCl were significantly reduced compared with the 5 mM KF treatment. The results point to a possible competitive inhibition of F transport by Cl. We also suggest that the possibility that aquaporins may be involved in the transmembrane transport of F should be further investigated. PMID- 19121100 TI - Photosynthesis, non-photochemical pathways and activities of antioxidant enzymes in a resilient evergreen oak under different climatic conditions from a valley savanna in Southwest China. AB - Plants in the savanna-valleys in Southwest China are annually exposed to different combinations of multiple stresses from the hot-rainy, to chill-dry, and to warm-dry seasons. This study monitored seasonal changes in photosynthesis and photoprotection in an evergreen oak (Cyclobalanopsis helferiana) from one of these valleys for four years during which usual and abnormal drought occurred. In general, during the study period with decreasing xylem water potential (Psix), photosynthetic gas exchange, quantum yield of photosystem II (PSII) photochemistry and activities of most of the measured antioxidant enzymes decreased, while activities of the xanthophyll cycle and associated non photochemical energy dissipation and glutathione peroxidase (GP) (EC 1.11.1.9) increased. In a fairly severe chill period, high concentration of reactive oxygen species induced high activities of most of the antioxidant enzymes and relatively stronger decrease in gas exchange. In the most severe dry period, even when predawn Psix decreased down to -4 MPa, considerable Pn (maximum photosynthetic rate) (4 micromol m(-2) s(-1)) was still maintained in midmorning. At this time, most of the antioxidant enzyme activities decreased to the lowest values, whereas the xanthophyll cycle and associated non-photochemical energy dissipation and GP activities increased to their highest levels. High predawn antheraxanthin and zeaxanthin contents were observed in the severe and very severe drought periods. Superoxide dismutase maintained high and fairly constant activity (1500-1800 U mg(-1) protein) and predawn maximum photochemistry efficiency of PSII was always above 0.8 throughout the whole study period. These results indicated that the photosynthetic apparatus of the oak leaves was highly capable of maintaining its function under the multiple stresses in different seasons in the present valley savanna. PMID- 19121101 TI - Relative abundance of Delta(5)-sterols in plasma membrane lipids of root-tip cells correlates with aluminum tolerance of rice. AB - We investigated variations in aluminum (Al) tolerance among rice plants, using ancestor cultivars from the family line of the Al-tolerant and widely cultivated Japonica cultivar, Sasanishiki. The cultivar Rikuu-20 was Al sensitive, whereas a closely related cultivar that is a descendant of Rikuu-20, Rikuu-132, was Al tolerant. These two cultivars were compared to determine mechanisms underlying variations in Al tolerance. The sensitive cultivar Rikuu-20 showed increased permeability of the plasma membrane (PM) and greater Al uptake within 1 h of Al treatment. This could not be explained by organic acid release. Lipid composition of the PM differed between these cultivars, and may account for the difference in Al tolerance. The tolerant cultivar Rikuu-132 had a lower ratio of phospholipids to Delta(5)-sterols than the sensitive cultivar Rikuu-20, suggesting that the PM of Rikuu-132 is less negatively charged and less permeabilized than that of Rikuu 20. We used inhibitors of Delta(5)-sterol synthesis to alter the ratio of phospholipids to Delta(5)-sterols in both cultivars. These inhibitors reduced Al tolerance in Rikuu-132 and its Al-tolerant ancestor cultivars Kamenoo and Kyoku. In addition, Rikuu-132 showed a similar level of Al sensitivity when the ratio of phospholipids to Delta(5)-sterols was increased to match that of Rikuu-20 after treatment with uniconazole-P, an inhibitor of obtusifoliol-14alpha-demethylase. These results indicate that PM lipid composition is a factor underlying variations in Al tolerance among rice cultivars. PMID- 19121102 TI - Functional differentiation of the sugar beet root system as indicator of developmental phase change. AB - Developmental phase transitions in the plant root system have not been well characterized. In this study we compared the dynamics of sucrose accumulation with changes in gene expression analyzed with cDNA-amplified fragment length polymorphism (AFLP) in the developing tap root of sugar beet (Beta vulgaris, L.) during the first 9 weeks after emergence (WAE). Although differences between lines were evident as soon as 9 WAE, sucrose showed a marked increase in the rate of accumulation between 4 and 6 WAE and a remarkable shift in gene expression was observed between 5 and 6 WAE. These changes were evident in two unrelated genetic backgrounds and suggest that physiological and gene expression changes represent a functional differentiation of the tap root. These changes were considered as indicators of a developmental change in the sugar beet root system. To identify genes and metabolic pathways involved in this developmental shift, a root cDNA library was hybridized with probes enriched for 3- and 7-WAE transcripts and differentially expressed transcripts were analyzed by cDNA microarray. Several genes involved in the regulation of tissue development were found to be differentially regulated. Genes involved in protein metabolism, disease-related and secretory system were upregulated before the functional differentiation transition, while genes under hormonal control were upregulated after the functional differentiation transition. This developmental phase change of the root system is important to understand plant developmental regulation at the whole-plant level and will likely be useful as early selection parameter in breeding programs. PMID- 19121104 TI - Welcome to the first issue of the seventh volume of the Plant Biotechnology Journal.. PMID- 19121103 TI - Involvement of actin filaments in rhizoid morphogenesis of Spirogyra. AB - The role of actin filaments in rhizoid morphogenesis was studied in Spirogyra. When the algal filaments were severed, new terminal cells started tip growth and finally formed rhizoids. Actin inhibitors, latrunculin B and cytochalasin D, reversibly inhibited the process. A mesh-like structure of actin filaments (AFs) was formed at the tip region. Gd(3+) inhibited tip growth and decreased AFs in the tip region. Either a decrease in turgor pressure or lowering of the external Ca(2+) concentration also induced similar results. It was suggested that the mesh like AF structure is indispensable for the elongation of rhizoids. A possible organization mechanism of the mesh-like AF structure was discussed. PMID- 19121105 TI - Resetting and regulation of Flowering Locus C expression during Arabidopsis reproductive development. AB - The epigenetic regulation of the floral repressor Flowering Locus C (FLC) is one of the critical factors that determine flowering time in Arabidopsis thaliana. Although many FLC regulators, and their effects on FLC chromatin, have been extensively studied, the epigenetic resetting of FLC has not yet been thoroughly characterized. Here, we investigate the FLC expression during gametogenesis and embryogenesis using FLC::GUS transgenic plants and RNA analysis. Regardless of the epigenetic state in adult plants, FLC expression disappeared in gametophytes. Subsequently, FLC expression was reactivated after fertilization in embryos, but not in the endosperm. Both parental alleles contributed equally to the expression of FLC in embryos. Surprisingly, the reactivation of FLC in early embryos was independent of FRIGIDA (FRI) and SUPPRESSOR OF FRIGIDA 4 (SUF4) activities. Instead, FRI, SUF4 and autonomous-pathway genes determined the level of FLC expression only in late embryogenesis. Many FLC regulators exhibited expression patterns similar to that of FLC, indicating potential roles in FLC reprogramming. An FVE mutation caused ectopic expression of FLC in the endosperm. A mutation in PHOTOPERIOD-INDEPENDENT EARLY FLOWERING 1 caused defects in FLC reactivation in early embryogenesis, and maintenance of full FLC expression in late embryogenesis. We also show that the polycomb group complex components, Fertilization-Independent endosperm and MEDEA, which mediate epigenetic regulation in seeds, are not relevant for FLC reprogramming. Based on our results, we propose that FLC reprogramming is composed of three phases: (i) repression in gametogenesis, (ii) reactivation in early embryogenesis and (iii) maintenance in late embryogenesis. PMID- 19121106 TI - NRAMP genes function in Arabidopsis thaliana resistance to Erwinia chrysanthemi infection. AB - AtNRAMP3 and AtNRAMP4 are two Arabidopsis metal transporters sharing about 50% sequence identity with mouse NRAMP1. The NRAMP1/Slc11A1 metal ion transporter plays a crucial role in the innate immunity of animal macrophages targeted by intracellular bacterial pathogens. AtNRAMP3 and AtNRAMP4 localize to the vacuolar membrane. We found that AtNRAMP3 is upregulated in leaves challenged with the bacterial pathogens Pseudomonas syringae and Erwinia chrysanthemi, whereas AtNRAMP4 expression is not modified. Using single and double nramp3 and nramp4 mutants, as well as lines ectopically expressing either of these genes, we show that AtNRAMP3 and, to a lesser extent, AtNRAMP4 are involved in Arabidopsis thaliana resistance against the bacterial pathogen E. chrysanthemi. The susceptibility of the double nramp3 nramp4 mutant is associated with the reduced accumulation of reactive oxygen species and ferritin (AtFER1), an iron storage protein known to participate in A. thaliana defense. Interestingly, roots from infected plants accumulated transcripts of AtNRAMP3 as well as the iron deficiency markers IRT1 and FRO2. This finding suggests the existence of a shoot to-root signal reminiscent of an iron-deficiency signal activated by pathogen infection. Our data indicate that the functions of NRAMP proteins in innate immunity have been conserved between animals and plants. PMID- 19121107 TI - Gibberellin controls the nodulation signaling pathway in Lotus japonicus. AB - Root nodule formation is regulated by several plant hormones, but the details of the regulation of the nodulation signaling pathway are largely unknown. In this study, the role of gibberellin (GA) in the control of root nodule symbiosis was investigated at the physiological and genetic levels in Lotus japonicus. Exogenous application of biologically active GA, GA(3), inhibited the formation of infection threads and nodules, which was counteracted by the application of a biosynthesis inhibitor of GA, Uniconazole P. Nod factor-induced root hair deformation was severely blocked in the presence of GA, which was phenocopied by nsp2 mutants. The number of spontaneous nodules triggered by the gain-of-function mutation of calcium/calmodulin-dependent kinase (CCaMK) or the lotus histidine kinase 1 (LHK1) was decreased upon the addition of GA; moreover, the overexpression of the gain-of-function mutation of L. japonicus, SLEEPY1, a positive regulator of GA signaling, resulted in a reduced nodule number, without other aspects of root development being affected. These results indicate that higher GA signaling levels specifically inhibit the nodulation signaling pathway. Nod factor-dependent induction of NSP2 and NIN was inhibited by exogenous GA. Furthermore, the cytokinin-dependent induction of NIN was suppressed by GA. From these results, we conclude that GA inhibits the nodulation signaling pathway downstream of cytokinin, possibly at NSP2, which is required for Nod factor dependent NIN expression. These results clarify the roles of GA in the nodulation signaling pathway, and in relation to the cytokinin signaling pathway for nodulation in L. japonicus. PMID- 19121108 TI - How plants cope with biotic interactions. AB - In their natural environment, plants interact with many different organisms. The nature of these interactions may range from positive, for example interactions with pollinators, to negative, such as interactions with pathogens and herbivores. In this special issue, the contributors provide several examples of how plants manage both positive and negative biotic interactions. This review aims to relate their findings to what we know about the complex natural environments in which plants have evolved. Molecular analyses of plant genomes and expression profiles have shown how intricately plants may regulate responses to single or multiple biotic interactions. Plant responses are fine-tuned by signalling hormone interactions. When multiple organisms interact with a single plant this may result in antagonistic or synergistic effects. The emerging fields of ecogenomics and metabolomics undoubtedly will refine our understanding of the multilayered regulation that plants use to manage relationships with their biotic environment. However, we can only understand why plants have such an intricate regulatory apparatus if we consider the ecological context of plant biotic interactions. PMID- 19121109 TI - Pollinators, flowering phenology and floral longevity in two Mediterranean Aristolochia species, with a review of flower visitor records for the genus. AB - The pollination of Aristolochia involves the temporary confinement of visitors inside the flower. A literature review has shown that some species are visited by one or a few dipteran families, while others are visited by a wider variety of dipterans, but only some of these are effective pollinators. We observed flowering phenology and temporal patterns of pollinator attendance in diverse populations of Aristolochia baetica and A. paucinervis, two species that grow in SW Spain, frequently in mixed populations. The two species had overlapping floral phenologies, extended flowering periods and long-lived flowers. A. baetica attracted a higher number of visitors than A. paucinervis. Drosophilids and, to a lesser extent, phorids, were the main pollinators of A. baetica, whereas in A. paucinervis, phorids were the only pollinators. Attendance to A. paucinervis flowers by phorids in mixed populations was markedly lower than in pure populations. This effect was more evident in years with lower pollinator density. Our results suggest that A. baetica and A. paucinervis may compete for pollinators in mixed populations. PMID- 19121110 TI - Pollination by deceit in Paphiopedilum barbigerum (Orchidaceae): a staminode exploits the innate colour preferences of hoverflies (Syrphidae). AB - Paphiopedilum barbigerum T. Tang et F. T. Wang, a slipper orchid native to southwest China and northern Vietnam, produces deceptive flowers that are self compatible but incapable of mechanical self-pollination (autogamy). The flowers are visited by females of Allograpta javana and Episyrphus balteatus (Syrphidae) that disperse the orchid's massulate pollen onto the receptive stigmas. Measurements of insect bodies and floral architecture show that the physical dimensions of these two fly species correlate with the relative positions of the receptive stigma and dehiscent anthers of P. barbigerum. These hoverflies land on the slippery centralised wart located on the shiny yellow staminode and then fall backwards through the labellum entrance. They are temporarily trapped in the inflated chamber composed of the interconnected labellum and column. The attractive staminode of P. barbigerum strongly reflects the colour yellow (500 560 nm), a colour preferred innately by most pollen-eating members of the Syrphidae. No scent molecules were detected using GC mass spectrometry analysis, showing that the primary attractant in this system is visual, not olfactory. Pollination-by-deceit in P. barbigerum is contrasted with its congener, P. dianthum, a brood site mimic that is pollinated by ovipositing females of E. balteatus. As the natural rate of fruit set in P. barbigerum (mean 26.3% pooled over three seasons) is lower than that of P. dianthum (mean 58.5% over two seasons), the evolution of false brood sites in some Paphiopedilum spp. should be selectively advantageous as they may provide an increase in the attention and return rates of dependable pollinators to flowers that always lack a reward. PMID- 19121111 TI - Perfume-collecting male euglossine bees as pollinators of a basal angiosperm: the case of Unonopsis stipitata (Annonaceae). AB - Pollination of Unonopsis stipitata (Annonaceae) by males of two perfume collecting bees, Euglossa imperialis and Eulaema bombiformis (Euglossini) is described. This is the first detailed account of this pollination mode in a member of a basal angiosperm family. Pollinator behaviour, identification of the odour bouquet and electrophysiological reaction of one of the two pollinators to the odour bouquet were determined. The collected odour is produced by 'osmophores' located adaxially on the petals. Starch and polysaccharides accumulated in petals are metabolized during odour emission. Mainly monoterpenes were detected in the scent samples, among them trans-carvone oxide. This molecule is thought by several authors to be the key attractant for male Eulaema bees and may be pivotal for convergent evolution of the perfume-collecting syndrome among dicotyledonous and monocotyledonous plants. It is speculated that Unonopsis, which on the basis of molecular age dating is considered a relatively recent genus of the Annonaceae (being 15-30 million years old), has diversified in relation to male euglossine bee pollinators. PMID- 19121112 TI - Florivory increases selfing: an experimental study in the wild strawberry, Fragaria virginiana. AB - Florivores are antagonists that damage flowers, and have direct negative effects on flowering and pollination of the attacked plants. While florivory has mainly been studied for its consequences on seed production or siring success, little is known about its impact on mating systems. Damage to flowers can alter pollinator attraction to the plant and may therefore modify patterns of pollen transfer. However, the consequences of damage for mating systems can take two forms: a decrease in flower number reduces opportunities for intra-inflorescence pollen deposition (geitonogamy), which, in turn, may lead to a decrease in selfing; whereas a decrease in floral display may also reduce overall visitation and thus increase the chances of self-pollination via facilitated or autonomous autogamy. We investigated the effects of damage by a bud-clipping weevil (Anthonomus signatus) in Fragaria virginiana in an experimental setting mimicking natural conditions. We found that increased damage led to an increase in selfing, a result consistent with the increased autogamy pathway. We discuss the implications of this finding and evaluate the generality of florivore-mediated mating system expression. PMID- 19121113 TI - The interaction between Cistaceae and a highly specific seed-harvester ant in a Mediterranean scrubland. AB - We studied the interaction between the ant Goniomma kugleri and Cistaceae in a Cistus ladanifer-dominated scrubland, in southwestern Spain. We monitored seed harvesting, and studied ant preferences among Cistaceae seeds and their capture efficiencies for preferred seeds. For the stand of C. ladanifer, we estimated seed losses due to the ants. Harvesting was restricted to two seasons: mid-autumn to late winter, and late spring. Ant diet relied on Cistaceae seeds: during autumn and winter 90% of seeds returned to nests were of C. ladanifer, and the remaining fraction also comprised Cistaceae seeds. At this time, the ants harvested seeds directly from the plants. In late spring, the ant diet consisted of Tuberaria guttata s.l. seeds. Goniomma kugleri selectively collected Cistaceae seeds. For preferred species, seed removal rates at the colony level and seed capture times invested by individual workers were correlated with seed size. Because of shorter capture time and higher success frequency, capture efficiency in terms of number of seeds captured per unit time was higher for small-seeded species. Although each ant colony collected large numbers (up to 10(5)) of C. ladanifer seeds over the autumn-winter season, the impact of ant removal on the annual seed output was moderate, at around 20%. It is likely that, in C. ladanifer, the staggered seed release period, and the pulsed exposure of seed clumps in capsules through progressive locule dehiscence, effectively minimise seed losses to the ants. PMID- 19121114 TI - Phytophthora infestans isolates from Northern China show high virulence diversity but low genotypic diversity. AB - Phenotypic and genotypic characteristics of 48 Phytophthora infestans isolates, collected in five provinces in Northern China between 1997 and 2003, were determined and compared with reference isolates. Characterisation included mating type, virulence, mitochondrial DNA (mtDNA) haplotype and DNA fingerprinting patterns based on simple sequence repeats (SSR) and amplified fragment length polymorphisms (AFLP). All isolates had the A1 mating type, mtDNA haplotype IIa and an identical SSR genotype (designated as SG-01-01) that differed from SSR genotypes found in the reference isolates, including those representing the 'old' US-1 lineage that dominated the P. infestans population worldwide prior to 1980. In contrast, the virulence spectra were highly variable and virulence to all resistance genes present in the standard differential set (R1 to R11) was found. AFLP analysis revealed some diversity; eight different AFLP genotypes were found that could be grouped into two major clusters. This study shows that there is very little genotypic diversity in the P. infestans population in Northern China. The occurrence of many different races within this rather uniform population is discussed in the framework of recent insights into the molecular determinants of avirulence in potato-P. infestans'gene-for-gene' interactions. PMID- 19121115 TI - Resistance in pepper plants induced by Fusarium oxysporum f. sp. lycopersici involves different defence-related genes. AB - Inoculation with Fusarium oxysporum f. sp. lycopersici (FOL) protects pepper plants from subsequent infection with Phytophthora capsici. In the present paper, the level of local and systemic protection achieved by plants induced with FOL was evaluated by quantifying the pathogen biomass and using real-time PCR. Differences in the amount of pathogen were found in stems and roots between FOL treated and untreated plants, while pathogen biomass could not be detected in leaves of induced plants. Five defence-related genes coding for a PR-1 protein, a beta-1,3-glucanase, a chitinase, a peroxidase and a sesquiterpene cyclase were up regulated 48 h after treatment in all the tissues studied, and maximal mRNAs levels were found in leaves. PMID- 19121116 TI - Changes in C12:0, C18:1, C18:2 and C20:2 fatty acid content in wheat treated with resistance inducers and infected by powdery mildew. AB - This work presents a global investigation of total fatty acid (FA) content in wheat in relation to treatment with four inducers of resistance and to powdery mildew infection. Linolenic acid (C18:3), linoleic acid (C18:2) and palmitic acid (16:0) were the most abundant FAs in wheat leaves. We investigated the effect of the following inducers of resistance: Iodus40, heptanoyl salicylic acid (HSA), Milsana and trehalose on FA accumulation. Previous studies established that lipid metabolism is altered by these compounds, and we therefore aimed to characterise their impact at the FA level. During a time course experiment, content (quantitative analysis) and percentage (qualitative analysis) of FAs were compared in treated plants and in controls, as well as in plants inoculated with Blumeria graminis f. sp. tritici (i) and non-inoculated (ni) plants. No change in C18:3 content was observed. C18:1 in Iodus 40-treated (ni) plants showed a quantitative 1.2-fold increase. Lauric acid (C12:0) content quantitatively increased after Iodus 40 (2.8-fold), Milsana (4.8-fold) and trehalose (4.0-fold) treatment in (i) plants. However, eicosadienoic acid (C20:2) quantitatively decreased in (ni) plants after Iodus 40 (1.5-fold) and Milsana (2.3-fold) treatment. The amount of C18:2 increased (1.6-fold) after HSA treatment in (i) plants. All these variations in FA content were correlated with variations in the corresponding relative percentages. Our work provides the first evidence for alterations in C12:0, C18:1, C18:2 and C20:2 FA content caused by four resistance inducers. We also compared the amount and percentage of each FA in untreated (i) and (ni) plants. In (i) plants, eicosadienoic acid (C20:2) increased and C18:2 decreased slightly. The potential involvement of these FAs during induced resistance and infection is discussed. PMID- 19121117 TI - Suppression of SERK gene expression affects fungus tolerance and somatic embryogenesis in transgenic lettuce. AB - The Somatic embryogenesis receptor-like kinase (SERK) gene plays an important role in plant somatic and zygotic embryogenesis induction. The gene encodes an LRR-containing receptor-like kinase protein. Studies have been carried out focusing on different aspects of its function, but definitive conclusions on its role are far from being reached. SERK expression is generally detected in cells in which somatic or zygotic embryogenesis has been triggered. Transgenic lettuce lines were produced to silence the endogenous SERK gene using antisense RNA. The average number of seeds per flower in the R(1) and R(2) generations was similar for both transgenic and non-transgenic lines. However, a reduction in the number of viable grained seeds was observed in four studied transgenic lines. Endogenous SERK expression analysis revealed the absence of detectable LsSERK gene transcripts in three transgenic lines, which presented a reduction in their ability to form in vitro somatic embryonic structures. In addition, transgenic lines showed enhanced susceptibility to the pathogenic fungus Sclerotinia sclerotiorum, when compared to control plants. The results support the idea that SERK genes might not only be involved in plant growth and development, but probably also in a general mechanism of biotic and abiotic stress perception. PMID- 19121118 TI - MYB72, a node of convergence in induced systemic resistance triggered by a fungal and a bacterial beneficial microbe. AB - Colonisation of plant roots by selected beneficial Trichoderma fungi or Pseudomonas bacteria can result in the activation of a systemic defence response that is effective against a broad spectrum of pathogens. In Arabidopsis thaliana, induced systemic resistance (ISR) triggered by the rhizobacterial strain Pseudomonas fluorescens WCS417r is regulated by a jasmonic acid- and ethylene dependent defence signalling pathway. Jasmonic acid and ethylene also play a role in Trichoderma-induced resistance. To further investigate the similarities between rhizobacteria- and Trichoderma-induced resistance, we studied the response of Arabidopsis to root colonisation by Trichoderma asperellum T34. In many aspects T34-ISR was similar to WCS417r-ISR. First, colonisation of the roots by T34 rendered the leaves more resistant to the bacterial pathogen Pseudomonas syringae pv. tomato, the biotrophic oomycete Hyaloperonospora parasitica and the necrotrophic fungus Plectosphaerella cucumerina. Second, treatment of the roots with T34 primed the leaf tissue for enhanced jasmonic acid-responsive gene expression and increased formation of callose-containing papillae upon pathogen attack. Third, T34-ISR was fully expressed in the salicylic acid impaired mutant sid2, but blocked in the defence regulatory mutant npr1. Finally, we show that the root-specific transcription factor MYB72, which is essential in early signalling steps of WCS417r-ISR, is also required for T34-ISR. Together, these results indicate that the defence pathways triggered by beneficial Trichoderma and Pseudomonas spp. strains are highly similar and that MYB72 functions as an early node of convergence in the signalling pathways that are induced by these different beneficial microorganisms. PMID- 19121119 TI - Induction of systemic resistance in Arabidopsis thaliana in response to a culture filtrate from a plant growth-promoting fungus, Phoma sp. GS8-3. AB - The plant growth-promoting fungus (PGPF), Phoma sp. GS8-3, isolated from a zoysia grass rhizosphere, is capable of protecting cucumber plants against virulent pathogens. This fungus was investigated in terms of the underlying mechanisms and ability to elicit systemic resistance in Arabidopsis thaliana. Root treatment of Arabidopsis plants with a culture filtrate (CF) from Phoma sp. GS8-3 elicited systemic resistance against the bacterial speck pathogen Pseudomonas syringae pv. tomato DC3000 (Pst), with restricted disease development and inhibited pathogen proliferation. Pathway-specific mutant plants, such as jar1 (jasmonic acid insensitive) and ein2 (ethylene insensitive), and transgenic NahG plants (impaired in salicylate signalling) were protected after application of the CF, demonstrating that these pathways are dispensable (at least individually) in CF mediated resistance. Similarly, NPR1 interference in npr1 mutants had no effect on CF-induced resistance. Gene expression studies revealed that CF treatment stimulated the systemic expression of both the SA-inducible PR-1 and JA/ET inducible PDF1.2 genes. However, pathogenic challenge to CF-treated plants was associated with potentiated expression of the PR-1 gene and down-regulated expression of the PDF1.2 gene. The observed down-regulation of the PDF1.2 gene in CF-treated plants indicates that there may be cross-talk between SA- and JA/ET dependent signalling pathways during the pathogenic infection process. In conclusion, our data suggest that CF of Phoma sp. GS8-3 induces resistance in Arabidopsis in a manner where SA and JA/ET may play a role in defence signalling. PMID- 19121120 TI - Phylogenetic and transcriptional analysis of a strictosidine synthase-like gene family in Arabidopsis thaliana reveals involvement in plant defence responses. AB - Protein domains with similarity to plant strictosidine synthase-like (SSL) sequences have been uncovered in the genomes of all multicellular organisms sequenced so far and are known to play a role in animal immune responses. Among several distinct groups of Arabidopsis thaliana SSL sequences, four genes (AtSSL4 AtSSL7) arranged in tandem on chromosome 3 show more similarity to SSL genes from Drosophila melanogaster and Caenorhabditis elegans than to other Arabidopsis SSL genes. To examine whether any of the four AtSSL genes are immune-inducible, we analysed the expression of each of the four AtSSL genes after exposure to microbial pathogens, wounding and plant defence elicitors using real-time quantitative RT-PCR, Northern blot hybridisation and Western blot analysis with antibodies raised against recombinant AtSSL proteins. While the AtSSL4 gene was constitutively expressed and not significantly induced by any treatment, the other three AtSSL genes were induced to various degrees by plant defence signalling compounds, such as salicylic acid, methyl jasmonate and ethylene, as well as by wounding and exposure to the plant pathogens Alternaria brassicicola and cucumber mosaic virus. Our data demonstrate that the four SSL-coding genes are regulated individually, suggesting specific roles in basal (SSL4) and inducible (SSL5-7) plant defence mechanisms. PMID- 19121123 TI - The source of enhanced cognitive control in bilinguals: evidence from bimodal bilinguals. AB - Bilinguals often outperform monolinguals on nonverbal tasks that require resolving conflict from competing alternatives. The regular need to select a target language is argued to enhance executive control. We investigated whether this enhancement stems from a general effect of bilingualism (the representation of two languages) or from a modality constraint that forces language selection. Bimodal bilinguals can, but do not always, sign and speak at the same time. Their two languages involve distinct motor and perceptual systems, leading to weaker demands on language control. We compared the performance of 15 monolinguals, 15 bimodal bilinguals, and 15 unimodal bilinguals on a set of flanker tasks. There were no group differences in accuracy, but unimodal bilinguals were faster than the other groups; bimodal bilinguals did not differ from monolinguals. These results trace the bilingual advantage in cognitive control to the unimodal bilingual's experience controlling two languages in the same modality. PMID- 19121124 TI - The cognitive benefits of interacting with nature. AB - We compare the restorative effects on cognitive functioning of interactions with natural versus urban environments. Attention restoration theory (ART) provides an analysis of the kinds of environments that lead to improvements in directed attention abilities. Nature, which is filled with intriguing stimuli, modestly grabs attention in a bottom-up fashion, allowing top-down directed-attention abilities a chance to replenish. Unlike natural environments, urban environments are filled with stimulation that captures attention dramatically and additionally requires directed attention (e.g., to avoid being hit by a car), making them less restorative. We present two experiments that show that walking in nature or viewing pictures of nature can improve directed-attention abilities as measured with a backwards digit-span task and the Attention Network Task, thus validating attention restoration theory. PMID- 19121125 TI - When the truth is not too hard to handle: an event-related potential study on the pragmatics of negation. AB - Our brains rapidly map incoming language onto what we hold to be true. Yet there are claims that such integration and verification processes are delayed in sentences containing negation words like not. However, studies have often confounded whether a statement is true and whether it is a natural thing to say during normal communication. In an event-related potential (ERP) experiment, we aimed to disentangle effects of truth value and pragmatic licensing on the comprehension of affirmative and negated real-world statements. As in affirmative sentences, false words elicited a larger N400 ERP than did true words in pragmatically licensed negated sentences (e.g., "In moderation, drinking red wine isn't bad/good..."), whereas true and false words elicited similar responses in unlicensed negated sentences (e.g., "A baby bunny's fur isn't very hard/soft..."). These results suggest that negation poses no principled obstacle for readers to immediately relate incoming words to what they hold to be true. PMID- 19121126 TI - With a clean conscience: cleanliness reduces the severity of moral judgments. AB - Theories of moral judgment have long emphasized reasoning and conscious thought while downplaying the role of intuitive and contextual influences. However, recent research has demonstrated that incidental feelings of disgust can influence moral judgments and make them more severe. This study involved two experiments demonstrating that the reverse effect can occur when the notion of physical purity is made salient, thus making moral judgments less severe. After having the cognitive concept of cleanliness activated (Experiment 1) or after physically cleansing themselves after experiencing disgust (Experiment 2), participants found certain moral actions to be less wrong than did participants who had not been exposed to a cleanliness manipulation. The findings support the idea that moral judgment can be driven by intuitive processes, rather than deliberate reasoning. One of those intuitions appears to be physical purity, because it has a strong connection to moral purity. PMID- 19121127 TI - Differential processing of consonants and vowels in lexical access through reading. AB - Do consonants and vowels have the same importance during reading? Recently, it has been proposed that consonants play a more important role than vowels for language acquisition and adult speech processing. This proposal has started receiving developmental support from studies showing that infants are better at processing specific consonantal than vocalic information while learning new words. This proposal also received support from adult speech processing. In our study, we directly investigated the relative contributions of consonants and vowels to lexical access while reading by using a visual masked-priming lexical decision task. Test items were presented following four different primes: identity (e.g., for the word joli, joli), unrelated (vabu), consonant-related (jalu), and vowel-related (vobi). Priming was found for the identity and consonant-related conditions, but not for the vowel-related condition. These results establish the privileged role of consonants during lexical access while reading. PMID- 19121128 TI - Preference invites categorization. AB - Past research indicates that positive affect (relative to neutral or negative affect) reduces processing and makes categorization less differentiated. The present experiment demonstrated that preference, even though affectively pleasant, invites finer categorization. Expertise is already known to influence categorization; hence, the present experiment used an associative conditioning task (novel symbols paired with positively or negatively valenced photographs) to create new preferences, thereby demonstrating that preference influences categorization independently of preexisting expertise. These findings cast new light on established theory of affect and cognitive processing and suggest new implications for consumer preference and goal pursuit. PMID- 19121129 TI - An in-group advantage in detecting intergroup anxiety. AB - We examined the possibility of an in-group advantage in detecting intergroup anxiety. Specifically, we videotaped White and Black participants while they engaged in same-race or interrace interactions. Then we asked White and Black observers to view these videotapes (unaware of the racial context) and provide their impressions of participants' anxiety. Two results pointed to an in-group advantage in detecting intergroup anxiety. First, only same-race observers perceived a modulation of participants' anxious behavior as a function of racial context. This held true not only for relatively subjective perceptions of global anxiety, but also for perceptions of single, discrete behaviors tied to anxiety. Second, we found that only same-race observers provided descriptions of anxiety that tracked reliably with participants' cortisol changes during the task. These results suggest that White and Black Americans may have difficulty developing a sense of shared emotional experience. PMID- 19121130 TI - Looking to the future to appreciate the present: the benefits of perceived temporal scarcity. AB - Both psychological research and conventional wisdom suggest that it can be difficult to attend to and derive enjoyment from the pleasant things in life. The present study examined whether focusing on the imminent ending of a positive life experience can lead to increased enjoyment. A temporal distance manipulation was used to make college graduation seem more or less close at hand. Twice a week over the course of 2 weeks, college students were told to write about their college life, with graduation being framed as either very close or very far off. As predicted, thinking about graduation as being close led to a significant increase in college-related behaviors and subjective well-being over the course of the study. The present research provides support for the counterintuitive hypothesis that thinking about an experience's ending can enhance one's present enjoyment of it. PMID- 19121131 TI - Gender recognition of human faces using color. AB - A continuing question in the object recognition literature is whether surface properties play a role in visual representation and recognition. Here, we examined the use of color as a cue in facial gender recognition by applying a version of reverse correlation to face categorization in CIE L*a*b* color space. We found that observers exploited color information to classify ambiguous signals embedded in chromatic noise. The method also allowed us to identify the specific spatial locations and the components of color used by observers. Although the color patterns found with human observers did not accurately mirror objective natural color differences, they suggest sensitivity to the contrast between the main features and the rest of the face. Overall, the results provide evidence that observers encode and can use the local color properties of faces, in particular, in tasks in which color provides diagnostic information and the availability of other cues is reduced. PMID- 19121132 TI - Twelve-month-old infants benefit from prior experience in statistical learning. AB - A decade of research suggests that infants readily detect patterns in their environment, but it is unclear how such learning changes with experience. We tested how prior experience influences sensitivity to statistical regularities in an artificial language. Although 12-month-old infants learn adjacent relationships between word categories, they do not track nonadjacent relationships until 15 months. We asked whether 12-month-old infants could generalize experience with adjacent dependencies to nonadjacent ones. Infants were familiarized to an artificial language either containing or lacking adjacent dependencies between word categories and were subsequently habituated to novel nonadjacent dependencies. Prior experience with adjacent dependencies resulted in enhanced learning of the nonadjacent dependencies. Female infants showed better discrimination than males did, which is consistent with earlier reported sex differences in verbal memory capacity. The findings suggest that prior experience can bootstrap infants' learning of difficult language structure and that learning mechanisms are powerfully affected by experience. PMID- 19121133 TI - An attentional mechanism for selecting appropriate actions afforded by graspable objects. AB - An object may afford a number of different actions. In this article, we show that an attentional mechanism inhibits competing motor programs that could elicit erroneous actions. Participants made a speeded key press to categorize the second of two successively presented door handles that were rotated at varying orientations relative to one another. Their responding hand was compatible or incompatible with the graspable part of the door handles (rightward or leftward facing). Compatible responses were faster than incompatible responses if the two handles shared an identical orientation, but they were slower if the two handles were aligned at slightly dissimilar orientations. Such suppressive surround effects are hallmarks of attentional processing in the visual domain, but they have never been observed behaviorally in the motor domain. This finding delineates a common mechanism involved in two of the most important functions of the brain: processing sensory data and preparing actions based on that information. PMID- 19121134 TI - Powerful people make good decisions even when they consciously think. PMID- 19121135 TI - The sting of intentional pain. PMID- 19121136 TI - Is relative pitch specific to pitch? AB - Melodies, speech, and other stimuli that vary in pitch are processed largely in terms of the relative pitch differences between sounds. Relative representations permit recognition of pitch patterns despite variations in overall pitch level between instruments or speakers. A key component of relative pitch is the sequence of pitch increases and decreases from note to note, known as the melodic contour. Here we report that contour representations are also produced by patterns in loudness and brightness (an aspect of timbre). The representations of contours in different dimensions evidently have much in common, as contours in one dimension can be readily recognized in other dimensions. Moreover, contours in loudness and brightness are nearly as useful as pitch contours for recognizing familiar melodies that are normally conveyed via pitch. Our results indicate that relative representations via contour extraction are a general feature of the auditory system, and may have a common central locus. PMID- 19121137 TI - The fragility of intergroup relations: divergent effects of delayed audiovisual feedback in intergroup and intragroup interaction. AB - Intergroup interactions between racial or ethnic majority and minority groups are often stressful for members of both groups; however, the dynamic processes that promote or alleviate tension in intergroup interaction remain poorly understood. Here we identify a behavioral mechanism-response delay-that can uniquely contribute to anxiety and promote disengagement from intergroup contact. Minimally acquainted White, Black, and Latino participants engaged in intergroup or intragroup dyadic conversation either in real time or with a subtle temporal disruption (1-s delay) in audiovisual feedback. Whereas intergroup dyads reported greater anxiety and less interest in contact after engaging in delayed conversation than after engaging in real-time conversation, intragroup dyads reported less anxiety in the delay condition than they did after interacting in real time. These findings have theoretical and practical implications for understanding intergroup communication and social dynamics and for promoting positive intergroup contact. PMID- 19121138 TI - Reciprocity is not give and take: asymmetric reciprocity to positive and negative acts. AB - Unlike economic exchange, social exchange has no well-defined "value." It is based on the norm of reciprocity, in which giving and taking are to be repaid in equivalent measure. Although giving and taking are colloquially assumed to be equivalent actions, we demonstrate that they produce different patterns of reciprocity. In five experiments utilizing a dictator game, people reciprocated in like measure to apparently prosocial acts of giving, but reciprocated more selfishly to apparently antisocial acts of taking, even when the objective outcomes of the acts of giving and taking were identical. Additional results demonstrate that acts of giving in social exchanges are perceived as more generous than objectively identical acts of taking, that taking tends to escalate, and that the asymmetry in reciprocity is not due to gaining versus losing resources. Reciprocity appears to operate on an exchange rate that assigns value to the meaning of events, in a fashion that encourages prosocial exchanges. PMID- 19121139 TI - Category markers or attributes: why do labels guide infants' inductive inferences? AB - To clarify the role of labels in early induction, we compared 16-month-old infants' (n=114) generalization of target properties to test objects when objects were introduced by the experimenter in one of the following ways: (a) with a general attentional phrase, (b) highlighted with a flashlight and a general attentional phrase, (c) via a recorded voice that labeled the objects using a naming phrase, (d) with a label consisting of a count noun embedded within a naming phrase, (e) with a label consisting of a single word that was not marked as belonging to a particular grammatical form class, and (f) with a label consisting of an adjective. Infants relied on object labels to guide their inductive inferences only when the labels were presented referentially, embedded within an intentional naming phrase, and marked as count nouns. These results suggest that infants do not view labels as attributes of objects; rather, infants understand that count-noun labels are intentional markers denoting category membership. PMID- 19121140 TI - The case of the transmogrifying experimenter: affirmation of a moral schema following implicit change detection. AB - The meaning-maintenance model posits that threats to schemas lead people to affirm unrelated schemas. In two studies testing this hypothesis, participants who were presented with a perceptual anomaly (viz., the experimenter was switched without participants consciously noticing) demonstrated greater affirmation of moral beliefs compared with participants in a control condition. Another study investigated whether the schema affirmation was prompted by unconscious arousal. Participants witnessed the changing experimenter and then consumed a placebo. Those who were informed that the placebo caused side effects of arousal did not show the moral-belief affirmation observed in the previous studies, as they misattributed their arousal to the placebo. In contrast, those who were not informed of such side effects demonstrated moral-belief affirmation. The results demonstrate the functional interchangeability of different meaning frameworks, and highlight the role of unconscious arousal in prompting people to seek alternative schemas in the face of a meaning threat. PMID- 19121141 TI - Reorienting when cues conflict: evidence for an adaptive-combination view. AB - Proponents of a geometric module claim that human adults accomplish spatial reorientation in a fundamentally different way than young children and nonhuman animals do. However, reporting two experiments that used a conflict paradigm, this article shows striking similarities between human adults and young children, as well as nonhuman animals. Specifically, Experiment 1 demonstrates that adults favor geometric information in a small room and rely on features in a larger room, whereas Experiment 2 demonstrates that experience in a larger room produces dominance of features over geometric cues in a small room-the first human case of reliance on features that contradict geometric information. Thus, use of features during reorientation depends on the size of the environment and learning history. These results clearly undermine the modularity claim and the view that feature use during reorientation is purely associative, and we discuss the findings within an adaptive-combination view, according to which a weighting system determines use of feature or geometric cues during reorientation. PMID- 19121142 TI - Construal level and procrastination. AB - According to construal-level theory, events that are distant in time tend to be represented more abstractly than are events that are close in time. This mental association between level of abstractness and temporal distance is proposed to be a bidirectional relationship, such that level of representation of an event should also have effects on the time when the activity is performed. In the present studies, participants were asked to respond to a questionnaire via e-mail within 3 weeks. The questionnaire was designed to induce either an abstract or a concrete construal. Using a variety of manipulations of construal level, the studies supported the predictions of construal-level theory. Individuals were less likely to procrastinate performing the task when the questionnaire induced a more concrete construal. Furthermore, this effect did not depend on the attractiveness, importance, or perceived difficulty of the task. PMID- 19121143 TI - Power, distress, and compassion: turning a blind eye to the suffering of others. AB - Responses to individuals who suffer are a foundation of cooperative communities. On the basis of the approach/inhibition theory of power (Keltner, Gruenfeld, & Anderson, 2003), we hypothesized that elevated social power is associated with diminished reciprocal emotional responses to another person's suffering (feeling distress at another person's distress) and with diminished complementary emotion (e.g., compassion). In face-to-face conversations, participants disclosed experiences that had caused them suffering. As predicted, participants with a higher sense of power experienced less distress and less compassion and exhibited greater autonomic emotion regulation when confronted with another participant's suffering. Additional analyses revealed that these findings could not be attributed to power-related differences in baseline emotion or decoding accuracy, but were likely shaped by power-related differences in the motivation to affiliate. Implications for theorizing about power and the social functions of emotions are discussed. PMID- 19121144 TI - Transplant International. Editorial. PMID- 19121145 TI - Minimization of steroids in liver transplantation. AB - Because of the markedly improved short-term results of liver transplantation (LT) and persistently high number of long-term complications, the attention of transplant physicians should be focused on minimizing immunosuppressive therapy as much as possible. Steroid-based immunosuppression is responsible for a substantial post-LT morbidity and mortality, hence, minimization of its use is of utmost importance to improve the quality of life of the successfully transplanted liver recipient. This literature review shows that LT can be performed safely with steroid-minimal immunosuppression without compromising graft and patient survival. The tendency in clinical practice is to move more and more from steroid withdrawal to steroid avoidance protocols. PMID- 19121146 TI - Calcineurin inhibitor minimization protocols in liver transplantation. AB - Liver transplant recipients are at increasingly high risk for suffering from impaired renal function and probable need of renal replacement therapy. Extended criteria organs and transplantation of patients with higher model for end-stage liver disease scores further increase this problem. Acute and chronic nephrotoxicity are the trade-off in immunosuppression with potent calcineurin inhibitors (CNIs). As a good renal function is associated with better graft and patient survival, CNI minimization protocols have been developed. Current strategies to overcome CNI toxicity include reduction or withdrawal of CNIs concurrently with switching over to mammalian target of rapamycin inhibitor or mycophenolate mofetil (MMF)-based regimens. This strategy caused an improvement in renal function in a significant number of liver transplantation patients according to several studies. However, total CNI avoidance seems to result in higher rejection rates. To prevent chronic renal dysfunction in patients prone to or with acute renal failure, CNI delay - with induction therapy for bridging - followed by low-dose CNI in combination with MMF are proven strategies without risking higher rejection rates. An individualized, tailor-made immunosuppressive regime, with a special focus on renal function is recommended. This review gave an overview on CNI minimization protocols in liver transplantation also focusing on recently analyzed studies. PMID- 19121147 TI - Free distribution of insecticide treated bed nets to pregnant women in Kinshasa: an effective way to achieve 80% use by women and their newborns. AB - OBJECTIVE: To determine whether long lasting insecticide treated bed nets (LLINs) distributed free of charge to pregnant women at their first antenatal clinic visit in Kinshasa, DRC are used from the time of distribution to delivery and 6 months after delivery. METHODS: Women were enrolled into a cohort study at their first antenatal care (ANC) visit and provided LLINs free of charge. Reported use of these nets was then measured at the time of delivery (n = 328) and in a random sample of women (n = 100) 6 months post-delivery using an interviewer administered, structured questionnaire. RESULTS: At baseline, only 25% of women reported having slept under a bed net the night before the interview. At the time of delivery, after being provided an LLIN for free, this increased to 79%. Six months post-delivery (n = 100), 80% of women reported sleeping under a net with a child under the age of 5 the night before the interview. CONCLUSIONS: Freely distributed bed nets are acceptable, feasible and result in high usage. Free distribution of bed nets during antenatal clinic visits may be a highly effective way to rapidly increase the use of bed nets among both pregnant women and their newborn infants in areas with high levels of ANC attendance. PMID- 19121148 TI - The impact of primary health care on malaria morbidity--defining access by disease burden. AB - OBJECTIVES: Primary care facilities are increasingly becoming the focal point for distribution of malaria intervention strategies, but physical access to these facilities may limit the extent to which communities can be reached. To investigate the impact of travel time to primary care on the incidence of hospitalized malaria episodes in a rural district in Kenya. METHODS: The incidence of hospitalized malaria in a population under continuous demographic surveillance was recorded over 3 years. The time to travel to the nearest primary health care facility was calculated for every child between birth and 5 years of age and trends in incidence of hospitalized malaria as a function of travel time were evaluated. RESULTS: The incidence of hospitalized malaria more than doubled as travel time to the nearest primary care facility increased from 10 min to 2 h. Good access to primary health facilities may reduce the burden of disease by as much as 66%. CONCLUSIONS: Our results highlight both the potential of the primary health care system in reaching those most at risk and reducing the disease burden. Insufficient access is an important risk factor, one that may be inequitably distributed to the poorest households. PMID- 19121149 TI - Feasibility of satellite image-based sampling for a health survey among urban townships of Lusaka, Zambia. AB - OBJECTIVES: To describe our experience using satellite image-based sampling to conduct a health survey of children in an urban area of Lusaka, Zambia, as an approach to sampling when the population is poorly characterized by existing census data or maps. METHODS: Using a publicly available Quickbird image of several townships, we created digital records of structures within the residential urban study area using ArcGIS 9.2. Boundaries were drawn to create geographic subdivisions based on natural and man-made barriers (e.g. roads). Survey teams of biomedical research students and local community health workers followed a standard protocol to enroll children within the selected structure, or to move to the neighbouring structure if the selected structure was ineligible or refused enrollment. Spatial clustering was assessed using the K-difference function. RESULTS: Digital records of 16 105 structures within the study area were created. Of the 750 randomly selected structures, six (1%) were not found by the survey teams. A total of 1247 structures were assessed for eligibility, of which 691 eligible households were enrolled. The majority of enrolled households were the initially selected structures (51%) or the first selected neighbour (42%). Households that refused enrollment tended to cluster more than those which enrolled. CONCLUSIONS: Sampling from a satellite image was feasible in this urban African setting. Satellite images may be useful for public health surveillance in populations with inaccurate census data or maps and allow for spatial analyses such as identification of clustering among refusing households. PMID- 19121150 TI - Anthropometrically derived dosing and drug costing calculations for treating visceral leishmaniasis in Bihar, India. AB - OBJECTIVE AND METHOD: To estimate drug costs of treating visceral leishmaniasis (VL) based on data on the VL population structure from the high-burden, antimony resistant area of Northern Bihar, India. RESULTS: Paromomycin is the cheapest option ($7450 to treat 1000 patients). Treating 1000 patients with oral miltefosine would cost $119,250 at the current private market price or $64,383 $75,129 at preferential public sector price depending on the size of the order. With AmBisome it would be $163,600 or $229,500 depending on the dose (10 or 15 mg/kg total). These costs are without considering other direct costs (daily intramuscular injections for 3 weeks for paromomycin; intravenous devices and hospitalization for AmBisome; directly observed treatment if applied for miltefosine) and indirect costs. CONCLUSION: These calculations provide useful basic information for projections. PMID- 19121151 TI - Laminin receptor 1 is differentially expressed in thoracic and limb wounds in the horse. AB - Healing of wounds located on the distal limbs of horses is often complicated by retarded epithelialization and the development of exuberant granulation tissue (proud flesh). Treatments that definitively resolve this pathological process are still unavailable. Molecular studies of the repair mechanism might contribute to the development of new therapeutic strategies. The study presented herein aimed to clone the full length cDNA and to study the spatio-temporal expression profile of mRNA and protein for LAMR1, previously attributed a role in wound epithelialization, during the repair of body and limb wounds in the horse. Cloning was achieved by screening a cDNA library previously derived from 7-day wound biopsies. Expression was studied in unwounded skin and in samples from 1-, 2-, 3-, 4- and 6-week-old wounds of the body and limb. Temporal gene expression was determined by real time polymerase chain reaction (RT-PCR) while protein expression was mapped immunohistochemically. Full-length cDNA for equine LAMR1 was shown to be highly similar to that of other species. The mRNA expression of LAMR1 was significantly up-regulated only in thoracic wounds, 4 and 6 weeks following wounding (upon epithelialization). Cutaneous wounding induced protein expression at both locations. Our data suggest that up-regulation of LAMR1 protein might favour epithelialization during wound healing. However, its interaction with ligands other than laminin complicates data interpretation. Future studies should quantitatively verify the temporal expression of this protein in order to provide the basis for targeted therapies that might enhance epithelialization. PMID- 19121153 TI - Use of a wireless system to measure invasive arterial blood pressure in ponies - preliminary study. AB - OBJECTIVE: To evaluate the feasibility and functionality of intra-carotid wireless device implantation in ponies, and to investigate its short-term complications. STUDY DESIGN: Prospective preliminary study. ANIMALS: Five mixed breed, adult, intact male ponies weighing 104 +/- 28.8 kg (mean +/- SD) underwent surgery. Arterial blood pressure data were continuously collected from four animals. METHODS: General anesthesia was induced on two consecutive days. On the first day, an intra-arterial wireless device was implanted in the right carotid artery. On the next day, a transcutaneous intra-arterial catheter was placed in the left facial artery. Data from both sources were collected. Post-mortem examination was performed. RESULTS: Surgical time was 27.1 +/- 11.85 minutes. All catheters remained in place with some extra vascular migration. Complications included mild seroma and hematoma. CONCLUSION: The wireless system allowed continuous monitoring in ponies throughout anesthesia and at rest and may allow for the recording of arterial blood pressure and heart rate when it would be difficult to achieve with a conventional system (e.g. during recovery from anesthesia). CLINICAL RELEVANCE: The wireless invasive blood pressure monitor may allow continuous measurements when only intermittent measurements would be feasible with a wired system. PMID- 19121154 TI - Determination of the minimum alveolar concentration of isoflurane in Shetland ponies using constant current or constant voltage electrical stimulation. AB - OBJECTIVE: To determine the minimum alveolar concentration (MAC) of isoflurane in Shetland ponies using a sequence of three different supramaximal noxious stimulations at each tested concentration of isoflurane rather than a single stimulation. STUDY DESIGN: Prospective, experimental trial. ANIMALS: Seven 4-year old, gelding Shetland ponies. METHODS: The MAC of isoflurane was determined for each pony. Three different modes of electrical stimulation were applied consecutively (2 minute intervals): two using constant voltage (90 V) on the gingiva via needle- (CVneedle) or surface-electrodes (CVsurface) and one using constant current (CC; 40 mA) via surface electrodes applied to the skin over the digital nerve. The ability to clearly interpret the responses as positive, the latency of the evoked responses and the inter-electrode resistance were recorded for each stimulus. RESULTS: Individual isoflurane MAC (%) values ranged from 0.60 to 1.17 with a mean (+/-SD) of 0.97 (+/-0.17). The responses were more clearly interpreted with CC, but did not reach statistical significance. The CVsurface mode produced responses with a longer delay. The CVneedle mode was accompanied by variable inter-electrode resistances resulting in uncontrolled stimulus intensity. At 0.9 MAC, the third stimulation induced more positive responses than the first stimulation, independent of the mode of stimulation used. CONCLUSIONS: The MAC of isoflurane in the Shetland ponies was lower than expected with considerable variability among individuals. Constant current surface electrode stimulations were the most repeatable. A summation over the sequence of three supramaximal stimulations was observed around 0.9 MAC. CLINICAL RELEVANCE: The possibility that Shetland ponies require less isoflurane than horses needs further investigation. Constant current surface-electrode stimulations were the most repeatable. Repetitive supramaximal stimuli may have evoked movements at isoflurane concentrations that provide immobility when single supramaximal stimulation was applied. PMID- 19121155 TI - Effects of changing body position on oxygenation and arterial blood pressures in foals anesthetized with guaifenesin, ketamine, and xylazine. AB - OBJECTIVE: To investigate the impact of a change in body position on blood gases and arterial blood pressures in foals anesthetized with guaifenesin, ketamine, and xylazine. STUDY DESIGN: Prospective, randomized experimental study. ANIMALS: Twelve Quarter Horse foals, age of 5.4 +/-0.9 months and weighing 222 +/- 48 kg. METHODS: Foals were anesthetized with guaifenesin, ketamine, and xylazine for 40 minutes in lateral recumbency and then assigned to a change in lateral recumbency after hoisting (Group 1, n = 6), or no change (Group 2, n = 6). Oxygen 15 L minute(-1) was insufflated into the endotracheal tube throughout anesthesia. Arterial blood pressure, heart rate, respiratory rate (f(R)), inspired fraction of oxygen (FIO(2)), and end-tidal carbon dioxide (PE'CO(2)) were measured every 5 minutes. Arterial pH and blood gases [arterial partial pressure of oxygen (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2))] were measured at 10, 30, and 40 minutes after induction, and 5 minutes after hoisting. Alveolar dead space ventilation and PaO(2)/FIO(2) were calculated. Two repeated measures models were used. All hypothesis tests were two-sided and significance level was alpha = 0.05. All values are presented as least square means +/- SE. RESULTS: Values at time-matched points from the two groups were not significantly different so they were combined. Arterial partial pressure of oxygen decreased significantly from 149 +/- 14.4 mmHg before hoisting to 92 +/- 11.6 mmHg after hoisting (p = 0.0013). The PaO(2)/FIO(2) ratio decreased from 275 +/- 30 to 175 +/- 24 (p = 0.0055). End-tidal carbon dioxide decreased significantly from 48.7 +/- 1.6 to 44.5 +/- 1.2 mmHg (p = 0.021). Arterial partial pressure of carbon dioxide, blood pressures and heart rates measured 5 minutes after hoisting were not different from measurements obtained before hoisting. CONCLUSION AND CLINICAL RELEVANCE: Hoisting decreased PaO(2) in anesthetized healthy foals. Administration of supplemental oxygen is recommended to counter the decrease in oxygenation and PaO(2) measurement is necessary to detect early changes. PMID- 19121156 TI - Comparative study on the sedative effects of morphine, methadone, butorphanol or tramadol, in combination with acepromazine, in dogs. AB - OBJECTIVE: To compare the effects of morphine (MOR), methadone (MET), butorphanol (BUT) and tramadol (TRA), in combination with acepromazine, on sedation, cardiorespiratory variables, body temperature and incidence of emesis in dogs. STUDY DESIGN: Prospective randomized, blinded, experimental trial. ANIMALS: Six adult mixed-breed male dogs weighing 12.0 +/- 4.3 kg. METHODS: Dogs received intravenous administration (IV) of acepromazine (0.05 mg kg(-1)) and 15 minutes later, one of four opioids was randomly administered IV in a cross-over design, with at least 1-week intervals. Dogs then received MOR 0.5 mg kg(-1); MET 0.5 mg kg(-1); BUT 0.15 mg kg(-1); or TRA 2.0 mg kg(-1). Indirect systolic arterial pressure (SAP), heart rate (HR), respiratory rate (f(R)), rectal temperature, pedal withdrawal reflex and sedation were evaluated at regular intervals for 90 minutes. RESULTS: Acepromazine administration decreased SAP, HR and temperature and produced mild sedation. All opioids further decreased temperature and MOR, BUT and TRA were associated with further decreases in HR. Tramadol decreased SAP whereas BUT decreased f(R) compared with values before opioid administration. Retching was observed in five of six dogs and vomiting occurred in one dog in MOR, but not in any dog in the remaining treatments. Sedation scores were greater in MET followed by MOR and BUT. Tramadol was associated with minor changes in sedation produced by acepromazine alone. CONCLUSIONS AND CLINICAL RELEVANCE: When used with acepromazine, MET appears to provide better sedation than MOR, BUT and TRA. If vomiting is to be avoided, MET, BUT and TRA may be better options than MOR. PMID- 19121157 TI - Yohimbine antagonizes the anaesthetic effects of ketamine-xylazine in captive Indian wild felids. AB - OBJECTIVE: To determine the effectiveness of yohimbine as an antagonist of ketamine-xylazine anaesthesia in captive Asiatic lions (Panthera leo persica), tigers (Panthera tigris) and leopards (Panthera pardus). STUDY DESIGN: Prospective clinical trial. ANIMALS: Fifty-two healthy adult lions, 55 adult leopards and 16 adult male tigers. METHODS: Captive wild felids in Indian zoos were anaesthetized with a combination of ketamine (2.2-2.6 mg kg(-1)) and xylazine (1.1-1.3 mg kg(-1)) using a dart propelled from a blowpipe. Time to onset of anaesthesia, lateral recumbency and induction time were measured, and physiological variables (respiration, heart rate and rectal temperature) were recorded once after the onset of complete anaesthesia. Anaesthesia was antagonized at various time periods with an intravenous administration of either 0.1 or 0.15 mg kg(-1) yohimbine. Onset of arousal and time to complete anaesthetic recovery were recorded. RESULTS: A total of 123 immobilizations were conducted between 2000 and 2005. Anaesthetic induction was achieved in 15-25 minutes in all animals. Incidents of sudden recovery or life-threatening effects associated with immobilizations were not observed. Yohimbine effectively antagonized anaesthesia in all animals within 10 minutes without any excitatory behaviour compared to control animals. No adverse reactions/side effects to yohimbine were recorded except that a few leopards exhibited seizure-like signs for a short period immediately after yohimbine administration. The duration of anaesthesia had no significant effect on the recovery time in any of the animals. CONCLUSION AND CLINICAL RELEVANCE: Yohimbine antagonized the xylazine portion of ketamine-xylazine anaesthesia and thereby hastened recovery from anaesthesia in Asiatic lions, tigers and leopards. PMID- 19121158 TI - The cardiorespiratory and anesthetic effects of clinical and supraclinical doses of alfaxalone in cats. AB - OBJECTIVE: To determine the cardiorespiratory and anesthetic effects of 0, 5, 15, and 50 mg kg(-1) intravenous (IV) alfaxalone in hydroxypropyl beta cyclodextrin (Alfaxan; Jurox Pty Ltd, Rutherford, NSW, Australia) in cats. STUDY DESIGN: Four treatments of alfaxalone were administered in sequential order. ANIMALS: Eight healthy adult cats (four male; four female) weighing between 3.71 and 5.91 kg. METHODS: Cats were instrumented for hemodynamic measurements. Four (0, 5, 15, and 50 mg kg(-1)) IV doses of alfaxalone were administered over one minute, with a 3 hour washout period between doses 0, 5, and 15 mg kg(-1) on Day 0. The 50 mg kg( 1) treatment was administered 24 hours later. Measurements of heart rate, aortic systolic, mean, and diastolic blood pressures, pulmonary arterial and right atrial mean pressures, cardiac output, respiratory rate, tidal and minute volumes, and arterial blood pH and blood gases (PaO(2), PaCO(2)) were performed at pre-determined intervals. Systemic vascular resistance and rate pressure product were calculated. The quality of induction, maintenance, and recovery from anesthesia and the response to noxious stimulation were categorically scored. RESULTS: Alfaxalone administration resulted in dose-dependent cardiorespiratory depression. Decreases in arterial blood pressure and increases in heart rate occurred at higher doses. Most variables returned to baseline by 15-30 minutes. Respiratory rate, minute volume, and PaO(2) decreased. Apnea was the most common side effect. Induction and maintenance quality were judged to be good to excellent at all doses and quality of recovery good to excellent at all but the 50 mg kg(-1) dose. The duration of anesthesia and unresponsiveness to noxious stimulation increased with dose. The administration of the 50 mg kg(-1) dose produced marked cardiorespiratory depression and apnea. CONCLUSIONS AND CLINICAL RELEVANCE: Alfaxalone produced dose-dependent anesthesia, cardiorespiratory depression and unresponsiveness to noxious stimulation in unpremedicated cats. Hypoventilation and apnea were the most common side effects. PMID- 19121159 TI - The effect of volatile anaesthetics on the relative sensitivity of facial and distal thoracic limb muscles to vecuronium in dogs. AB - OBJECTIVE: To compare n. facialis-m. nasolabialis (nF-mNL) and n. ulnar-mm. carpi flexorii (nU-mCF) sensitivity to vecuronium during halothane or isoflurane anaesthesia. STUDY DESIGN: Randomized, prospective, experimental study. ANIMALS: Forty-four client-owned dogs (19 male, 25 female) undergoing surgery; mean age: 5.0 years; mean body mass: 24.7 kg. METHODS: Thirty minutes after acepromazine (0.05 mg kg(-1)) and morphine (0.5 mg kg(-1)), anaesthesia was induced with intravenous (IV) thiopental and maintained with either halothane (n = 22) or isoflurane (randomly allocated) in oxygen. The lungs were mechanically ventilated and end-tidal inhaled anaesthetic (FE'(IAA)) maintained at 1.2 x MAC values. Neuromuscular transmission at nF-uNL and nU-mCF was monitored using the train of four count. Vecuronium (50 microg kg(-1) IV) was injected (t = 0) after 15 trains, 50-60 minutes after inhalational anaesthesia began, when FE'(IAA) had been constant for >15 minutes. Times of the disappearance (-) and reappearance (+) of the fourth (T4) and first twitch (T1) were recorded allowing the calculation of: latent (t = 0 to T4-) and manifest onset times (t = 0 to T1-) duration of blockade (T1- to T1+) and drug effect (T4- to T4+) and recovery time (T1+-T4+). Student's paired t-test was used to compare simultaneous responses at nF-uNL and nU-mCF. An unpaired t-test was used to compare anaesthetic effects. RESULTS: Latent and manifest onset times were significantly (p < 0.05) briefer, blockade and drug effects were significantly longer and recovery from blockade were significantly slower in the nF-mNL unit in both halothane and isoflurane recipients. Profound block duration and drug action were significantly longer and recovery from blockade were significantly slower in halothane recipients at both nerve-muscle units. CONCLUSION AND CLINICAL RELEVANCE: The nF-mNL was more sensitive than nU-mCF to vecuronium, particularly in halothane-anaesthetized dogs. PMID- 19121160 TI - Brightness of venous blood in South American camelids: implications for jugular catheterization. AB - OBJECTIVE: To compare the brightness of South American camelid venous blood to that of Equidae. STUDY DESIGN: Prospective clinical evaluation. ANIMALS: Twelve South American camelids (eight llamas, four alpacas), eight horses and ponies (control group). METHODS: Appropriately sized catheters were placed in the jugular vein of each animal under local anaesthesia. The blood spilt before the catheter was capped was caught on a white tile. A sample of blood was drawn for blood-gas analysis. The brightness of the blood (both on the tile and in the syringe) was matched to a colour chart (1 = darkest red, 8 = brightest red) by a single observer under bright light conditions. Packed cell volume (PCV) and partial pressure of oxygen (PvO(2)) in the blood were also measured on the syringe blood. Normally distributed data were compared using a two tailed t-test, and non-normally distributed data were compared using a Mann-Whitney U-test. Significance was set at p < 0.05. RESULTS: Camelid venous blood was significantly brighter red than that of horses and ponies both on the white tile (p = 0.0003) and in the syringe (p = 0.0001). PCV was significantly lower in camelids (32 +/- 4%) compared with horses (37 +/- 5%). Partial pressure of oxygen values were similar between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Jugular venous blood in alpacas and llamas is significantly brighter red than that of horses. Colour should not be used as a sole determinant of venous or arterial catheterization in this species. PMID- 19121161 TI - A comparison of epidural buprenorphine plus detomidine with morphine plus detomidine in horses undergoing bilateral stifle arthroscopy. AB - OBJECTIVE: To compare the analgesic efficacy of buprenorphine plus detomidine with that of morphine plus detomidine when administered epidurally in horses undergoing bilateral stifle arthroscopy. STUDY DESIGN: Prospective, randomized, blinded clinical trial. ANIMALS: Twelve healthy adult horses participating in an orthopedic research study. Group M (n = 6) received morphine (0.2 mg kg(-1)) and detomidine (0.15 mg kg(-1)) epidurally; group B (n = 6) received buprenorphine (0.005 mg kg(-1)) and detomidine (0.15 mg kg(-1)) epidurally. METHODS: Horses received one of two epidural treatments following induction of general anesthesia for bilateral stifle arthroscopy. Heart rate (HR), mean arterial blood pressure (MAP), end-tidal CO(2) (Pe'CO(2)), and end-tidal isoflurane concentrations (E'Iso%) were recorded every 15 minutes following epidural administration. Post operative assessment was performed at 1, 2, 3, 6, 9, 12, and 24 hours after standing; variables recorded included HR, respiratory rate (f(R)), abdominal borborygmi, defecation, and the presence of undesirable side effects. At the same times post-operatively, each horse was videotaped at a walk and subsequently assigned a lameness score (0-4) by three ACVS diplomates blinded to treatment and who followed previously published guidelines. Nonparametric data were analyzed using Wilcoxon's rank-sum test. Inter- and intra-rater agreement were determined using weighted kappa coefficients. Statistical significance was set at p 1 year) returned to their intended use (37 brood mares, 2 racehorses, and 1 show horse) with no chronic problems related to the surgical procedure. CONCLUSION: None of the variables examined were associated with survival. Outcomes were similar to other large studies of surgical colic in the horse. Self-limiting diarrhea is common after large colon resection and the prognosis for survival after hospital discharge is favorable. CLINICAL RELEVANCE: Horses that survive the early postoperative period and are discharged after large colon resection and anastomosis have a good chance for long-term survival with minimal negative impact on quality of life and use. PMID- 19121176 TI - Urinary tract infection in dogs with thoracolumbar intervertebral disc herniation and urinary bladder dysfunction managed by manual expression, indwelling catheterization or intermittent catheterization. AB - OBJECTIVE: To evaluate risk factors for lower urinary tract infection (UTI) in dogs with intervertebral disc disease (IVDD) that had manual expression (ME), indwelling catheterization (IDC) or intermittent catheterization (ITC) for urinary bladder management. STUDY DESIGN: Randomized-clinical trial. ANIMALS: Dogs (n=62) treated with urinary bladder dysfunction requiring surgery for IVDD and control dogs (n=30) that had surgery for reasons other than IVDD. METHODS: Treated dogs were randomly assigned to ME, IDC, or ITC. Urine was collected for culture and antimicrobial susceptibility testing before and after treatment. Incidence and risk factors for UTI were evaluated. Bacterial isolates and antimicrobial resistance patterns were described. RESULTS: Mean (+/-SD) time to urination was significantly longer for IDC dogs (7.4+/-2.75 days) than ME dogs (4.2+/-2.63) and ITC dogs (4.9+/-3.12). Thirteen treated dogs (21%) and no control dogs developed UTI: 4/25 (16%) ME, 8/25 (32%) IDC, and 1/12 (8%) ITC. Enterobacter sp. was most frequently isolated (4/13; 31%). Duration of treatment was the only risk factor for UTI and each additional day of treatment increased the risk of UTI 1.5 times. CONCLUSION: For dogs with acute IVDD, the duration of required urinary bladder management establishes the risk of UTI, not the urinary bladder management technique. CLINICAL RELEVANCE: Duration of treatment for urinary bladder dysfunction is a risk factor for UTI in dogs recovering from acute IVDD. Treatment for urinary bladder management should be limited where possible and no method of treatment is preferred. For dogs managed by IDC, voluntary urination might occur before clinically suspected. PMID- 19121177 TI - Spinal epidural abscess in two calves. AB - OBJECTIVE: To report clinical signs, diagnostic and surgical or necropsy findings, and outcome in 2 calves with spinal epidural abscess (SEA). STUDY DESIGN: Clinical report. ANIMALS: Calves (n=2). METHODS: Calves had neurologic examination, analysis and antimicrobial culture of cerebrospinal fluid (CSF), vertebral column radiographs, myelography, and in 1 calf, magnetic resonance imaging (MRI). A definitive diagnosis of SEA was confirmed by necropsy in 1 calf and during surgery and histologic examination of vertebral canal tissue in 1 calf. RESULTS: Clinical signs were difficulty in rising, ataxia, fever, apparent spinal pain, hypoesthesia, and paresis/plegia which appeared 15 days before admission. Calf 1 had pelvic limb weakness and difficulty standing and calf 2 had severe ataxia involving both thoracic and pelvic limbs. Extradural spinal cord compression was identified by myelography. SEA suspected in calf 1 with discospondylitis was confirmed at necropsy whereas calf 2 had MRI identification of the lesion and was successfully decompressed by laminectomy and SEA excision. Both calves had peripheral neutrophilia and calf 2 had neutrophilic pleocytosis in CSF. Bacteria were not isolated from CSF, from the surgical site or during necropsy. Calf 2 improved neurologically and had a good long-term outcome. CONCLUSION: Good outcome in a calf with SEA was obtained after adequate surgical decompression and antibiotic administration. CLINICAL RELEVANCE: SEA should be included in the list of possible causes of fever, apparent spinal pain, and signs of myelopathy in calves. PMID- 19121178 TI - Long-term outcome in dogs after circumcostal gastropexy or gastrocolopexy for gastric dilatation with or without volvulus. PMID- 19121180 TI - Rationale for randomized controlled trials and for intention-to-treat analysis in transfusion medicine: are they one and the same? AB - Articles on the appropriateness of intention-to-treat (ITT) vs. as-treated (AT) analyses of randomized controlled trials (RCTs) have highlighted issues relevant to drug trials, such as approaches suitable for 'explanatory' vs. 'pragmatic' RCTs. These considerations are less relevant to transfusion medicine RCTs, especially those of red blood cell transfusion therapies where the main issue is whether to include in the analysis randomized patients who did not receive transfusion. This article discusses issues pertinent specifically to transfusion medicine RCTs, and the thesis presented here is that the primary analysis of any transfusion medicine RCT must be based on the ITT principle. This is because the rationale for randomization is to avoid selection bias and to generate experimental arms that are comparable with respect to all, known and unknown, confounding factors. Only the entire arms produced by random allocation of subjects are so comparable, and only an ITT analysis of these entire arms fulfills the purpose of randomization. Withdrawals of randomized patients compromise the comparability of the groups and the rationale for randomization. Deviations from the ITT principle may be valid only when other conditions are met to ensure that non-adherence to ITT will not bias the results. For RCTs of red blood cell transfusion therapies, such conditions include that the RCTs be double blind and that transfusion criteria should be applied consistently. Nonetheless, the rationale for ITT can be reversed in equivalence and non-inferiority trials where the finding of no difference is the objective of the research; thus, both ITT and AT analyses should be presented in these settings. PMID- 19121181 TI - A new method of concentrating hepatitis B virus (HBV) DNA and HBV surface antigen: an application of the method to the detection of occult HBV infection. AB - BACKGROUND: The risk of post-transfusion hepatitis B virus (HBV) infection has been reduced after the implementation of HBV nucleic acid amplification technology (NAT). However, the problem of HBV DNA-positive and HBV surface antigen (HBsAg)-negative occult HBV infections remains to be solved. This is in part due to the HBV DNA load being too low to detect these occult HBV infections using mini-pool NAT. In Japan, the assay for the antibody against the HBV core antigen (anti-HBc) has not completely excluded occult HBV infection. To solve this problem, we have developed a new method of concentrating HBV DNA and HBsAg simultaneously to increase the sensitivity of detection tests. METHODS: Virus concentration is achieved by the enhancement of the agglutination of viruses using poly-L-lysine in the presence of a bivalent metal. Poly-L-lysine-coated magnetic beads are used to shorten the time of each step of the concentration procedure. Seventy-seven anti-HBc-positive and HBsAg-negative donations were examined. HBsAg and anti-HBc were tested by enzyme immunoassay (EIA) (AxSYM; Abbott) and haemagglutination inhibition test (Japanese Red Cross), respectively. RESULTS: HBV surface antigen and HBV DNA levels were concentrated up to four- to sevenfold. Using this method, 35 of the 77 anti-HBc-positive and HBsAg-negative donors were HBV DNA-positive by individual NAT and a further five donors became HBV DNA-positive by HBV concentration. Twenty-seven of 40 occult HBV infections became HBsAg-positive by HBsAg concentration. CONCLUSION: Our new method of concentrating HBV and HBsAg increased the sensitivities of EIA and HBV NAT, and enabled us to detect 27 of 40 occult HBV infections by HBsAg EIA. PMID- 19121182 TI - Sterility screening of platelet concentrates: questioning the optimal test strategy. AB - BACKGROUND: Routine bacterial monitoring of apheresis platelet concentrates (APC) and pooled platelet concentrates (PPC) was introduced in two German blood services using culture and real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). The results of testing are reviewed and used to discuss different strategies for detection of bacterial contamination of PCs. STUDY DESIGN AND METHODS: Two thousand three hundred and sixty-two APCs and 1993 PPCs have been tested by real-time RT-PCR and the BacT/Alert automated culturing system using aerobic and anaerobic culture bottles. After standard processing of PCs and storage of 22-24 h at 20-24 degrees C with agitation, samples were taken under aseptic conditions. Reactive culture bottles were confirmed as positive and bacterial isolates were identified by 16S rRNA analysis and biochemical tests. RESULTS: Seventeen of 2362 tested APCs were reactive in culture and one also in RT-PCR. Of these, 13 APCs were identified as initially positive as Staphylococcus warneri (n = 1, positive in aerobic and anaerobic culture), Propionibacterium acnes (n = 12, positive only in anaerobic culture) and four were initially reactive. Two of 1993 PPCs were initially reactive (anaerobic) and two more were confirmed positive (anaerobic) from a repeat culture and identified as P. acnes. All remaining specimens were tested negative. CONCLUSION: Our study demonstrates that the predominant organisms implicated in platelet bacterial contamination are part of the human skin flora. Inoculating blood culture systems and anaerobic cultivation detects these bacteria after approximately 3-7 days when blood products have been transfused. Based on the presented data different screening strategies are discussed. PMID- 19121183 TI - Trans-sialidase inhibition assay for the detection of Trypanosoma cruzi infection in blood donor samples from Argentina. AB - BACKGROUND AND OBJECTIVES: Conventional serology tests for Trypanosoma cruzi blood banks screening are neither sensitive nor specific enough, and currently no gold standard assay is available. Trans-sialidase inhibition assay (TIA) detects neutralizing antibodies against T. cruzi trans-sialidase. Conventional serology inconclusive, positive and negative blood donor samples were evaluated by employing TIA as a supplementary test. MATERIALS AND METHODS: Three hundred and twenty-one blood donor samples were tested using a combination of assays. Based on the results of testing, these were divided into a number of groups. All samples were tested by TIA. RESULTS: In conventional serology inconclusive samples 48.1% were TIA-positive, 1/54 conventional serology positive samples was TIA-negative. All negative samples from donors without epidemiological risks were TIA-negative; 1/48 was positive in those with epidemiological risk. CONCLUSION: Trans-sialidase inhibition assay application in blood banks may be useful to resolve inconclusive samples, and thus improves donor counseling and allows individual re-entry. The use of TIA in samples from negative conventional test donors but positive epidemiological antecedents may contribute to decrease transfusional risk. PMID- 19121184 TI - The characterization and impact of microparticles on haemostasis within fresh frozen plasma. AB - BACKGROUND: We have previously demonstrated that clot formation in fresh-frozen plasma (FFP) is influenced by the presence of microparticles (MP). In this study, the cellular source(s), properties and influence of MPs on clot formation within FFP were further characterized. METHODS: Fresh-frozen plasma was prepared after an overnight hold of whole blood at 4 degrees C. We examined the effect of a 0.2 microm filtration device designed to remove cellular MPs on thrombin generation test (TGT) and Thrombelastography (TEG(R)) as well as clotting factors and physiological inhibitors: prothrombin time (PT); activated partial thromboplastin time (APTT), fibrinogen (Fg), factor VIII (FVIII), von Willebrand factor antigen (VWF:Ag), antithrombin III (AT-III) and protein C (PC). MPs were measured using a functional assay and also by flow cytometry. RESULTS: Microparticle levels by functional assay were reduced by filtration (pre- 5.11 vs. post- 4.43 nmol/l phosphatidylserine equivalent, P < 0.0001). Flow cytometry showed that the most numerous MPs were derived from red blood cells, with ~87% binding annexin V, most of which (94%) were removed by filtration. MP removal had minimal effect on the PT, APTT, Fg, VWF:Ag, AT-III or PC or FVIII, but a major effect on TGT (endogenous thrombin potential: pre- 1722 vs. post- 990 nM thrombin, P < 0.0001; peak thrombin: pre- 91 vs. post- 44 nM thrombin, P < 0.0001), which in turn reflected the changes seen in TEG(R), where post-filtration clots started forming more slowly and the rate of clot formation was reduced. CONCLUSION: These data suggest that MPs contribute towards clot formation in FFP. PMID- 19121185 TI - Rationalizing blood transfusion in cardiac surgery: the impact of a red cell volume-based guideline on blood usage and clinical outcome. AB - BACKGROUND AND OBJECTIVES: Cardiac surgery is currently considered one of the heaviest users of red blood cells. An explanation may be found, in part, in considering the effect of the heavy clear fluid load associated with cardiopulmonary bypass. This may result in the artificial depression of haemoglobin concentration, overestimating the requirement for red cell transfusion if this is the sole parameter considered. To address this issue, we examined the impact of a red cell volume-based transfusion guideline on transfusion requirement. MATERIALS AND METHODS: This was a single-centre, randomized controlled trial. The cohort of 86 patients was allocated to receive red cells as per the red cell volume guideline (group RCV) or standard haemoglobin concentration-based departmental policy (group C). Outcome measures were red cell transfusion and clinical outcome. RESULTS: All preoperative data were comparable between the two groups. A significantly fewer percentage of patients in group RCV were transfused red cells (RCV = 32.6% vs. C = 53.5%, P = 0.05). No significant difference was found between any of the outcome measures with the exception of median hospital stay (RCV = 5.9 days vs. C = 6.8 days, P = 0.02). CONCLUSION: In elective cardiac surgery patients, considering haemoglobin concentration alone may overestimate the requirement for red cell transfusion. More research is required to determine the impact of restrictive transfusion policies on clinical outcome following cardiac surgery. PMID- 19121186 TI - An international collaborative study to establish a replacement World Health Organization International Standard for human immunodeficiency virus 1 RNA nucleic acid assays. AB - BACKGROUND AND OBJECTIVES: An international collaborative study was undertaken to identify a replacement for the World Health Organization (WHO) 1st International Standard for human immunodeficiency virus 1 (HIV-1) RNA for use in nucleic acid based techniques (NAT) (code 97/656). In the original study to establish the 1st International Standard, a second candidate material (code 97/650) had been shown to perform well and this was re-evaluated to establish whether it would be a suitable replacement. MATERIALS AND METHODS: Eight laboratories from six different countries participated in the collaborative study to evaluate the candidate replacement standard. A total of eight different NATs were used, five in a quantitative format and three qualitative, of which five were commercially available. RESULTS: The results showed that the estimates of RNA copies in the current study were generally in line with those of the original study and there was no evidence of any drift in overall levels expressed in International Units (IU) for the candidate standard between the two studies. Furthermore, it was shown to be stable over long-term storage at -20 degrees C. CONCLUSIONS: The candidate material code 97/650 was established by the WHO as the 2nd International Standard for HIV-1 RNA for use in NAT and assigned a unitage of 5.56 log(10) (363 078) IU/vial. PMID- 19121187 TI - Overview of revised measures to prevent malaria transmission by blood transfusion in France. AB - Plasmodial transmission by blood donation is rare in non-endemic countries, but a very serious complication of blood transfusion. The French national blood service (Etablissement Francais du Sang and Centre de Transfusion sanguine des Armees) intended to revise the measures to strengthen blood safety with regard to Plasmodiae as transmissible pathogens. To limit the risk of transmission during infusion, serious additive measures have been taken for more than a decade in France, which is the European country with the highest rate of exposure to imported plasmodial infections or malaria. These measures were revised and strengthened after the occurrence of a lethal transfusion-transmitted infection in 2002, but did not prevent another occurrence in 2006. This report examines the weaknesses of the systems and aims at emphasizing the safety measures already taken and addresses issues to best respond to that risk. PMID- 19121188 TI - Delayed intravascular haemolysis following multiple asymptomatic ABO-incompatible red blood cell transfusions in a patient with hepatic failure. AB - ABO-incompatible red blood cell (RBC) transfusions have rarely been associated with delayed haemolysis. However, we report the case of a 75-year-old man (blood type O) with hepatic disease, who received 5 units of incompatible type B RBCs over 8 days. The patient did not develop symptomatic or biochemical evidence of haemolysis until 7-8 days after the first incompatible RBC unit. The patient had a low anti-B antibody titre (1 : 64) prior to the first transfusion. The onset of haemolysis was temporally associated with an increase in anti-B and the infusion of fresh-frozen plasma. In conclusion, a patient with hepatic failure experienced a delayed haemolytic transfusion reaction after receiving multiple ABO incompatible RBC transfusions that were initially well-tolerated. We speculate that the delayed haemolysis may have resulted from an anamnestic antibody response to the initial incompatible transfusion, or possibly as a result of the transfusion of fresh-frozen plasma, which might have repleted low complement levels. PMID- 19121189 TI - Donors with a rare pheno (geno) type. PMID- 19121192 TI - Blood conservation strategies in major orthopaedic surgery: efficacy, safety and European regulations. AB - Several major orthopaedic surgical procedures may result in significant blood loss and the need for allogeneic blood transfusion (ABT). However, overall concerns about adverse effects of ABT have prompted the review of transfusion practice and the search for transfusion alternatives to decrease or avoid the use of ABT. These strategies include the correction of perioperative anaemia, pharmacological and non-pharmacologic measures to reduce blood loss, preoperative autologous blood donation and perioperative red blood cell salvage. We have reviewed the efficacy and safety of these strategies and where appropriate offer evidence-based recommendations on their use in orthopaedic surgery. We also reviewed the European regulations on ABT alternatives. Pharmacological alternatives need to be used with a total adherence to European regulations in their legal and off-label use. The administration and use of pharmacological agents to stimulate erythropoiesis or reduce blood loss needs to be within the context of attempting to use allogenic blood in a rational manner. As for autologous blood, European Directives cover preoperative autologous blood donation, but not its clinical use, and perioperative red blood cell salvage devices, but not the product yielded by them. Therefore, the development of quality standards and good practice guidelines for perioperative red blood cell salvage, as well as its inclusion in the haemovigilance programme, is urgently needed. Finally, it is noteworthy that some recommendations given for ABT alternatives are not supported by a high level of evidence and that the goal of performing major orthopaedic surgical procedures without the use of ABT may be better accomplished by combining several of these techniques within a defined algorithm. PMID- 19121193 TI - Inactivation of hepatitis A virus by heat and high hydrostatic pressure: variation among laboratory strains. AB - BACKGROUND AND OBJECTIVES: Hepatitis A virus (HAV) transmission via contaminated blood products has been reported. Cell-adapted HAV strains are generally used to confirm virus inactivation in manufacturing blood products, but the strains may differ in their sensitivity to inactivation treatment. To select an appropriate cell-adapted HAV strain for virus validation, we compared the inactivation efficiency among four strains under two different physical inactivation treatments: heat and high hydrostatic pressure. MATERIALS AND METHODS: The cell adapted HAV strains used here were KRM238, KRM003 (subgenotype IIIB), KRM031 (IA), and TKM005 (IB). The strains were treated at 60 degrees C for up to 10 h or under high hydrostatic pressure (up to 420 MPa). The reduction in HAV infectivity was measured by an immunofocus-staining method. RESULTS: The heat treatment at 60 degrees C for 10 h reduced HAV infectivity in the range of 3 to 5 log(10) among the strains; KRM238 and TKM005 were harder to inactivate than the other two. The high hydrostatic pressure treatment at 420 MPa also reduced infectivity in the range of 3 to 5 log(10) among the strains, and KRM031 was easier to inactivate than the other strains. CONCLUSION: Heat treatment and high hydrostatic pressure treatment revealed differences in inactivation efficiencies among cell-adapted HAV strains, and each strain reacted differently depending on the treatment. KRM238 may be the best candidate for virus validation to ensure the safety of blood products against viral contamination, as it is harder to inactivate and it replicates better in cell culture than the other strains. PMID- 19121194 TI - Predictors for non-compliance of notified hepatitis C virus-positive blood donors with recommendation to seek medical counselling. AB - BACKGROUND: Notification of blood donors represents the commonest method of informing asymptomatic individuals of abnormal test results indicating exposure to hepatitis C virus (HCV) infection. Such notification is therefore important from both health and economic perspectives. This study aimed to identify predictors for non-compliance of HCV-positive blood donors with the National Blood Services recommendation to seek medical counselling. STUDY DESIGN AND METHODS: The current research is a cross-sectional study. Telephone interviews were conducted with 201 blood donors identified as HCV positive following blood donation during 2001-2002 (40% response rate). RESULTS: About 25% of all the notified blood donors did not seek any counselling; 29% (44/150) of those who requested medical advice from their primary care physicians (general practitoner's) were not referred to specialists. Age, alcohol consumption and non practice of health-promoting behaviour were independent predictors of non compliance with the blood services' recommendation. In particular, smoking (odds ratio, 2.0; 95% confidence interval 1.0-4.2) and not undergoing professional teeth cleaning (odds ratio 2.8; 95% confidence interval 1.3-6.1) were found to be significant predictors of non-compliance. CONCLUSION: The study provides essential data regarding the extent and risk factors for non-compliance of HCV positive blood donors with recommendation to seek medical advice. Our results can assist in identifying blood donors who would not seek counselling, based on demographic factors and past exposure to risk factors for HCV. Improvements in the notification process and additional training of general practitoners regarding the management of HCV disease are needed. PMID- 19121195 TI - Efficacy of transfusion of platelet concentrates obtained by manual pooling or by semiautomated pooling of buffy-coats: a retrospective analysis of count increment, corrected count increment and transfusion interval. AB - BACKGROUND AND OBJECTIVE: The preparation of platelet concentrates from pooling buffy-coats can be done manually or semiautomatically with the OrbiSac BC System. The objective of this study was to compare the clinical outcome of platelet transfusions prepared by these two methods in terms of 1-h count increment (1-h CI), 1-h corrected count increment (1-h CCI) and transfusion interval. MATERIALS AND METHODS: Platelet transfusions were identified retrospectively for inclusion in the control group (manual pooling) or the test group (automated pooling). Transfusion outcome variables were 1-h CI, 1-h CCI and transfusion interval. The impact of 16 patient- and platelet concentrate-related variables on transfusion efficacy was investigated by multiple regression analysis. RESULTS: The database analysed consisted of 205 control transfusions given to 36 patients and 219 test transfusions given to 36 patients. Mean platelet content and mean 1-h CI were statistically higher in the test group when compared to the control group (P < 0.05), but mean 1-h CCI and transfusion interval were not. Multiple regression analysis resulted in models explaining 13% of the variance of 1-h CI, 8% of the variance of 1-h CCI and 17% of the variance of transfusion interval (P < 0.05). CONCLUSION: There are no significant differences between the two preparation methods regarding the clinical outcome of platelet transfusions, as the difference in 1-h CI is explained by differences in platelet content. PMID- 19121196 TI - Blood transfusion requirements and independent predictors of increased transfusion requirements among adult patients on extracorporeal membrane oxygenation -- a single centre experience. AB - BACKGROUND/OBJECTIVES: More adults undergo extracorporeal membrane oxygenation (ECMO) now. They have high transfusion requirements. This study described transfusion requirements of adults during ECMO in a single institution, and determined factors associated with high transfusion requirements. MATERIALS/METHODS: Retrospective analysis was done on the amount of blood products received by adults during ECMO. Predictors of increased average daily transfusion requirements during ECMO and increased ECMO duration (which correlated positively with total transfusion requirements) were determined. RESULTS: Forty-one patients (median age 50 years) underwent 42 ECMO sessions for respiratory failure (16.7%), cardiogenic shock (76.2%) or massive pulmonary embolism (7.1%). They received 569 red blood cells, 852 platelets, 126 fresh frozen plasma (FFP) and 220 cryoprecipitate in total during median ECMO duration of 5 (1-15) days. On multivariate analysis, average daily red blood cell transfusion increased with nadir haemoglobin (Hb) during ECMO (Hb(nadir)) of < 7.5 g/dl (P < 0.001). Average daily platelet transfusion increased with recent antiplatelet agents (P = 0.015) and maximum Hb decline of > 5.5 g/dl during ECMO (P = 0.011). Average daily platelet transfusion > 3 units was also associated with increased ECMO duration (P = 0.024). Average daily FFP transfusion was increased in patients with hypertension (P = 0.007) and Hb(nadir) < 7.5 g/dl (P = 0.050). Patients with sepsis (P = 0.009) or without surgery (P = 0.009) had increased ECMO duration, which correlated positively with total transfusion requirements during the entire ECMO session. ECMO improved mortality of patients with fulminant myocarditis, respiratory failure and massive pulmonary embolism. CONCLUSION: Adult ECMO patients with lower Hb(nadir) require more daily red blood cell and FFP. Hypertension increases daily FFP requirements. Recent antiplatelet agents, larger Hb decline and longer ECMO duration increase daily platelet requirements. Patients with sepsis or on ECMO for medical reasons have longer ECMO duration, which is associated with total transfusion requirements. Some of these factors may be identified early to optimize blood product support. PMID- 19121197 TI - Pilot study of oxygen transport rate of banked red blood cells. AB - BACKGROUND AND OBJECTIVES: Dynamic oximetry provides a new way to assess the effect of blood storage on the oxygen transport rate (OTR). MATERIALS AND METHODS: In dynamic oximetry, the rate at which oxyhemoglobin becomes deoxyhemoglobin is measured optically, thereby, indirectly measuring the rate at which oxygen leaves the red blood cell (RBC) making it available for transfer to tissues. Extending the physiologic diffusion time in an in vitro apparatus, consisting of a diffusion system and gas exchanger capable of controlling the surface area and the time of exposure for oxygenation and deoxygenation, makes OTR measurement feasible. Eight normal blood donor units, collected in adenine, dextrose, sorbitol, sodium chloride and mannitol , were stored for 8 weeks under standard conditions and serially sampled for OTR. RESULTS: We report that the OTR at the time of blood bank donation appears to be singular for each donor, that the interdonor differences are maintained over time, and that the individual OTR increased 1.72-fold (95% CI 1.51, 1.95) over 8 weeks, adjusting for sex, age and plasma cholesterol level. CONCLUSION: Oxygen transport rate increases during storage; blood units with similar haemoglobin content may have significant differences in OTR. Studies examining blood parameters at the time of donation and blood storage on patient outcomes should consider measuring OTR, as it may contribute to differences in observed efficacy of tissue oxygenation. PMID- 19121198 TI - Iron status in Norwegian blood donors: comparison of iron status in new blood donors registered in 1993-1997 and in 2005-2006. AB - BACKGROUND AND OBJECTIVES: The impact of a poor iron status on the difficulties to keep recruitment of new donors at pace with the ongoing increased demand for blood transfusions was studied by comparing the iron status of new donors recruited in 1993-1997 and in 2005-2006. MATERIALS AND METHODS: Iron status was defined by haemoglobin and serum ferritin. Inclusion criteria for approving new donors were haemoglobin >/= 12.5 g/dl for women and >/= 13.5 g/dl for men, and serum ferritin > 15 microg/l for both genders. Data were gathered retrospectively from 943 subjects (55% women) in the 1990 ties and prospectively from 1013 subjects (63% women) 10 years later. RESULTS: In women, there was a significant fall in haemoglobin and serum ferritin mean values from 13.2 to 13.1 g/dl and from 30.9 to 26.9 microg/l, respectively. Rejection due to low haemoglobin was significantly increased from 14% to 24%. In men, there were minor changes that did not affect rejection rates. CONCLUSION: Iron status of women who want to serve as blood donors has deteriorated in the last 10 years, leading to an increased rejection due to haemoglobin below the inclusion criterion for blood donors. PMID- 19121199 TI - Blood group A(1) and A(2) revisited: an immunochemical analysis. AB - BACKGROUND AND OBJECTIVE: The basis of blood group A(1) and A(2) phenotypes has been debated for many decades, and still the chemical basis is unresolved. The literature generally identifies the glycolipid chemical differences between blood group A(1) and A(2) phenotypes as being poor or no expression of A type 3 and A type 4 structures on A(2) red cells, although this assertion is not unanimous. MATERIALS AND METHODS: Using purified glycolipids and specific monoclonal antibodies, we revisited the glycolipid basis of the A(1) and A(2) phenotypes. Purified glycolipids were extracted from four individual A(1) and four individual A(2) blood units. One blood unit from an A weak subgroup was also included. Monoclonal anti-A reagents including those originally used to define the basis of A(1) and A(2) phenotypes were used in a thin layer chromatography - enzyme immunoassay to identify the presence of specific glycolipids. RESULTS: A type 3 glycolipid structures were found to be present in large amounts in all phenotypes. In contrast, the A type 4 glycolipid structure was virtually undetectable in the A(2) phenotype, but was present in the A(1) and A subgroup samples. CONCLUSION: The major glycolipid difference between the A(1) and A(2) phenotypes is the dominance of A type 4 glycolipids in the A(1) phenotype. PMID- 19121200 TI - Neonatal transfusions. PMID- 19121203 TI - Morbidity associated with "self-rated health" in epithelial ovarian cancer survivors. AB - BACKGROUND: Epithelial ovarian cancer survivors (EOCSs) frequently report multiple complaints after their treatment. The objective was to study somatic and mental morbidity in EOCSs associated with their Self-Rated Health (SRH) assessed by a single item.Findings were compared to age-matched controls from the general population. METHODS: In a cross -sectional follow-up design 189/287 (66%) EOCSs treated at The Norwegian Radiumhospital 1979-2003 responded to a mailed questionnaire on demographic data, and somatic and mental morbidity. SRH last week was rated on item #29 of the European Organization and Treatment of Cancer Quality of Life Questionnaire in 84/189 (97%) of responding EOCSs. For comparisons "good" and "poor" SRH groups were defined by the median score on the SRH item. RESULTS: EOCSs with "poor SRH" had higher level of somatic symptoms, anxiety, depression and fatigue than those with "good SRH" (p < .001). In multivariate analyses somatic symptoms, age and fatigue, were significantly associated with the SRH score in EOCSs, but not the cancer-related variables (FIGO stage, recurrence in < 6 months or chemotherapy ever). The model explained 70% of the variance in SRH in linear and 77% in logistic regression analyses. The distribution of the SRH scores in EOCSs did not differ significantly from that of normative controls; however a higher proportion of controls recorded a high SRH score. CONCLUSION: SRH is strongly related to common somatic complaints, impairment and fatigue but not to cancer-related variables. A single question concerning SRH last week might be a quick screening method for collecting important information on symptoms in EOCSs, in addition to cancer-related questions. PMID- 19121204 TI - Inhaled corticosteroids for asthma: impact of practice level device switching on asthma control. AB - BACKGROUND: As more inhaled corticosteroid (ICS) devices become available, there may be pressure for health-care providers to switch patients with asthma to cheaper inhaler devices. Our objective was to evaluate impact on asthma control of inhaler device switching without an accompanying consultation in general practice. METHODS: This 2-year retrospective matched cohort study used the UK General Practice Research Database to identify practices where ICS devices were changed without a consultation for > or =5 patients within 3 months. Patients 6 65 years of age from these practices whose ICS device was switched were individually matched with patients using the same ICS device who were not switched. Asthma control over 12 months after the switch was assessed using a composite measure including short-acting beta-agonist and oral corticosteroid use, hospitalizations, and subsequent changes to therapy. RESULTS: A total of 824 patients from 55 practices had a device switch and could be matched. Over half (53%) of device switches were from dry powder to metered-dose inhalers. Fewer patients in switched than matched cohort experienced successful treatment based on the composite measure (20% vs. 34%) and more experienced unsuccessful treatment (51% vs. 38%). After adjusting for possible baseline confounding factors, the odds ratio for treatment success in the switched cohort compared with controls was 0.29 (95% confidence interval [CI], 0.19 to 0.44; p < 0.001) and for unsuccessful treatment was 1.92 (95% CI, 1.47 to 2.56; p < 0.001). CONCLUSION: Switching ICS devices without a consultation was associated with worsened asthma control and is therefore inadvisable. PMID- 19121205 TI - Characteristics of chronic obstructive pulmonary disease in Spain from a gender perspective. AB - BACKGROUND: The objective of this study was to analyze the clinical and management characteristics of chronic obstructive pulmonary disease (COPD) in men and women, to determine possible gender-associated differences between the two groups of patients. METHODS: An observational and descriptive epidemiological study (EPIDEPOC study). The study included patients with stable COPD and aged >or= 40 years, evaluated in primary care. Data were collected relating to sociodemographic variables, clinical characteristics, quality of life (SF-12), severity of disease and treatment. The results obtained in men and women were compared. RESULTS: A total of 10,711 patients (75.6% males and 24.4% females) were evaluated. Significant differences were found between males and females in relation to the following parameters: age (67.4 +/- 9.2 years in men vs 66.1 +/- 10.8 in women, p < 0.05), smoking (91.9% of the men were smokers or ex-smokers vs 30% of the women), comorbidity (the frequency of hypertension, diabetes, anxiety and depression was greater in women, while ischemic heart disease was more common in men), mental component of quality of life (49.4 +/- 10.3 in men vs 44.6 +/- 11.9 in women, p < 0.05) and severity of disease (56.5 +/- 13.3% in men vs 60.7 +/- 3.2 in women, p < 0.05). As regards treatment, the percentage use of long acting b2-adrenergic agonists, anticholinergic agents, theophyllines and mucolytic agents was significant greater in men. The total annual cost of COPD was greater in males than in females (1989.20 +/- 2364.47 euro vs 1724.53 +/- 2106.90, p < 0.05). CONCLUSION: The women with COPD evaluated in this study were younger, smoked less and have more comorbidity, a poorer quality of life, and lesser disease severity than men with COPD. However, they generated a lesser total annual cost of COPD than men. PMID- 19121206 TI - An unusual cause of dysphagia in ductal breast cancer due to submucosal oropharyngeal metastatic spread: a case report. AB - INTRODUCTION: Invasive ductal and lobular carcinomas represent 67.9% and 6.3% of breast carcinoma, respectively. Metastatic breast cancer typically involves the lungs, bones, brain, and liver. Studies have shown differing patterns of metastatic spread between ductal and lobular carcinoma. Lobular carcinoma is more likely to metastasise to the gastrointestinal tract. CASE PRESENTATION: We report the case of a 49 year old white woman with invasive ductal carcinoma with lobular differentiation who developed submucosal oropharyngeal metastases nearly two years after her original diagnosis after presenting with odynophagia and dysphagia. The patient's symptoms preceded any associated radiological or endoscopic abnormalities by at least 9 months. Repeat computed tomography scan and eventual oropharyngeal biopsy confirmed submucosal metastatic invasive ductal carcinoma, suggesting occult submucosal spread. CONCLUSION: This case illustrates the importance of maintaining a high index of suspicion for metastatic disease in patients with invasive breast cancer who present with unusual symptoms and a careful search for metastatic sites. PMID- 19121207 TI - Antinuclear antibodies and their detection methods in diagnosis of connective tissue diseases: a journey revisited. AB - It has been more than 50 years since antinuclear antibodies were first discovered and found to be associated with connective tissue diseases. Since then different methods have been described and used for their detection or confirmation. For many decades immunofluorescent antinuclear antibody test has been the "gold standard" in the diagnosis of these disorders. However to increase the sensitivity and specificity of antinuclear antibody detection further approaches were explored. Today a battery of newer techniques are available some of which are now considered better and are competing with the older methods. This article provides an overview on advancement in antinuclear antibody detection methods, their future prospects, advantages, disadvantages and guidelines for use of these tests. PMID- 19121208 TI - Ophthalmic medicolegal cases in Upper Egypt. AB - PURPOSE: To describe the pattern of ophthalmic medicolegal cases with emphasis on cases of assault, and to acquaint ophthalmologists with rules pertaining to expert testimony and medical reports. METHODS: A retrospective study was carried out to review files of 247 medicolegal cases from Upper Egypt seen by the senior author in 8 years. These were classified categorically and were analyzed from various characteristics and aspects. The scheme for examination of subjects and for formulating the medicolegal report is described. RESULTS: The different categories were assault in 224 cases (90.5%), military recruitment evasion in 8 cases (3.25%), occupational disability claims in 8 cases (3.25%) and medical malpractice in 7 cases (3%). Thirty two cases (13%) presented with alleged functional visual loss, of them 25 cases (10%) were malingering. Traumatic lens subluxation or dislocation was seen in 37 (13.5%) cases and phthisis and atrophia bulbi was the presenting sign in 55 (22.3%) cases. Twenty percent of assault cases were females. There were no differences in incidence between the provinces in Upper Egypt. Assault tools inflicted injuries are described, as well as the outcome of these cases. Claims against military recruits could not be substantiated. Occupational claims for damages were false. Alleged medical negligence cases were rejected based on accepted standards of care and not on unexpected complications. CONCLUSION: Medical reports have to be structured, detailed, accurate and unbiased. Data in this work are useful for statistical and planning purposes in the medicolegal domain. PMID- 19121209 TI - A forensic aspect of age characteristics of dentine using transversal microradiography: a case report. AB - BACKGROUND: Translucency of dentine is the result of occlusion of the corresponding dentinal tubules by a mineral substance which has a refractive index similar to that of the rest of the dentine. CASE PRESENTATION: This case report describes the microradiographic features of an upper cadaveric canine. Transverse microradiograph is one of the methods assessing apical dentine translucency for various dental and medical reasons. CONCLUSION: Estimation of age using teeth structures may be of primary value in forensic dentistry, especially when soft tissues are severely destructed. PMID- 19121210 TI - Is U.S. health care an appropriate system? A strategic perspective from systems science. AB - CONTEXT: Systems science provides organizational principles supported by biologic findings that can be applied to any organization; any incongruence indicates an incomplete or an already failing system. U.S. health care is commonly referred to as a system that consumes an ever- increasing percentage of the gross domestic product and delivers seemingly diminishing value. OBJECTIVE: To perform a comparative study of U.S. health care with the principles of systems science and, if feasible, propose solutions. DESIGN: General systems theory provides the theoretical foundation for this observational research. MAIN OUTCOME MEASURES: A degree of compliance of U.S. health care with systems principles and its space time functional location within the dynamic systems model. RESULTS OF COMPARATIVE ANALYSIS: U.S. health care is an incomplete system further threatened by the fact that it functions in the zone of chaos within the dynamic systems model. CONCLUSION: Complying with systems science principles and the congruence of pertinent cycles, U.S. health care would likely dramatically improve its value creation for all of society as well as its resiliency and long-term sustainability.Immediate corrective steps could be taken: Prioritize and incentivize health over care; restore fiscal soundness by combining health and life insurance for the benefit of the insured and the payer; rebalance horizontal/providers and vertical/government hierarchies. PMID- 19121211 TI - The impact of obstructive sleep apnea variability measured in-lab versus in-home on sample size calculations. AB - BACKGROUND: When conducting a treatment intervention, it is assumed that variability associated with measurement of the disease can be controlled sufficiently to reasonably assess the outcome. In this study we investigate the variability of Apnea-Hypopnea Index obtained by polysomnography and by in-home portable recording in untreated mild to moderate obstructive sleep apnea (OSA) patients at a four- to six-month interval. METHODS: Thirty-seven adult patients serving as placebo controls underwent a baseline polysomnography and in-home sleep study followed by a second set of studies under the same conditions. The polysomnography studies were acquired and scored at three independent American Academy of Sleep Medicine accredited sleep laboratories. The in-home studies were acquired by the patient and scored using validated auto-scoring algorithms. The initial in-home study was conducted on average two months prior to the first polysomnography, the follow-up polysomnography and in-home studies were conducted approximately five to six months after the initial polysomnography. RESULTS: When comparing the test-retest Apnea-hypopnea Index (AHI) and apnea index (AI), the in home results were more highly correlated (r = 0.65 and 0.68) than the comparable PSG results (r = 0.56 and 0.58). The in-home results provided approximately 50% less test-retest variability than the comparable polysomnography AHI and AI values. Both the overall polysomnography AHI and AI showed a substantial bias toward increased severity upon retest (8 and 6 events/hr respectively) while the in-home bias was essentially zero. The in-home percentage of time supine showed a better correlation compared to polysomnography (r = 0.72 vs. 0.43). Patients biased toward more time supine during the initial polysomnography; no trends in time supine for in-home studies were noted. CONCLUSION: Night-to-night variability in sleep-disordered breathing can be a confounding factor in assessing treatment outcomes. The sample size of this study was small given the night-to-night variability in OSA and limited understanding of polysomnography reliability. We found that in-home studies provided a repeated measure of sleep disordered breathing less variable then polysomnography. Investigators using polysomnography to assess treatment outcomes should factor in the increased variability and bias toward increased AHI values upon retest to ensure the study is adequately powered. PMID- 19121212 TI - Establishment and characterization of three new human breast cancer cell lines derived from Chinese breast cancer tissues. AB - BACKGROUND: Breast cancer is a major malignancy affecting females worldwide. It is the most common cause of death from cancer in women. Cell lines are widely used in laboratory research and particularly as in vitro models in cancer research. But we found that the routinely used breast cancer cell lines were mostly derived from Caucasians or African-Americans. There were few standard models to study the pathogenic mechanism at molecular level and cell signaling pathway of breast cancer for Asian patients. It is quite necessary to establish new breast cancer cell lines from xanthoderm to study the pathogenic mechanism and therapeutic methods. RESULTS: Three new breast cancer cell lines, designated BC-019, BC-020 and BC-021, were successfully established and characterized from breast invasive ductal carcinoma tissues of three Chinese female patients. These new cell lines growing as adherent monolayer with characteristic epithelial morphology could be maintained continuously in vitro, and they were ER-, PR- and C-erbB-2-positive. Their chromosomes showed high hyperdiploidy and complex rearrangements, and they displayed aggressive tumorigencity in tumorigenesis test. CONCLUSION: The three newly established breast cancer cell lines from Chinese patients were tested for a number of, and the results indicate that the cell lines were in good quality and could be served as new cell models in breast cancer study. PMID- 19121213 TI - Phylogenetic relationships in Cortinarius, section Calochroi, inferred from nuclear DNA sequences. AB - BACKGROUND: Section Calochroi is one of the most species-rich lineages in the genus Cortinarius (Agaricales, Basidiomycota) and is widely distributed across boreo-nemoral areas, with some extensions into meridional zones. Previous phylogenetic studies of Calochroi (incl. section Fulvi) have been geographically restricted; therefore, phylogenetic and biogeographic relationships within this lineage at a global scale have been largely unknown. In this study, we obtained DNA sequences from a nearly complete taxon sampling of known species from Europe, Central America and North America. We inferred intra- and interspecific phylogenetic relationships as well as major morphological evolutionary trends within section Calochroi based on 576 ITS sequences, 230 ITS + 5.8S + D1/D2 sequences, and a combined dataset of ITS + 5.8S + D1/D2 and RPB1 sequences of a representative subsampling of 58 species. RESULTS: More than 100 species were identified by integrating DNA sequences with morphological, macrochemical and ecological data. Cortinarius section Calochroi was consistently resolved with high branch support into at least seven major lineages: Calochroi, Caroviolacei, Dibaphi, Elegantiores, Napi, Pseudoglaucopodes and Splendentes; whereas Rufoolivacei and Sulfurini appeared polyphyletic. A close relationship between Dibaphi, Elegantiores, Napi and Splendentes was consistently supported. Combinations of specific morphological, pigmentation and molecular characters appear useful in circumscribing clades. CONCLUSION: Our analyses demonstrate that Calochroi is an exclusively northern hemispheric lineage, where species follow their host trees throughout their natural ranges within and across continents. Results of this study contribute substantially to defining European species in this group and will help to either identify or to name new species occurring across the northern hemisphere. Major groupings are in partial agreement with earlier morphology-based and molecular phylogenetic hypotheses, but some relationships were unexpected, based on external morphology. In such cases, their true affinities appear to have been obscured by the repeated appearance of similar features among distantly related species. Therefore, further taxonomic studies are needed to evaluate the consistency of species concepts and interpretations of morphological features in a more global context. Reconstruction of ancestral states yielded two major evolutionary trends within section Calochroi: (1) the development of bright pigments evolved independently multiple times, and (2) the evolution of abruptly marginate to flattened stipe bulbs represents an autapomorphy of the Calochroi clade. PMID- 19121214 TI - Acute aortic dissection type A discloses Corpus alienum. AB - We report an unusual case of an aortic type A dissection with a corpus alienum which compresses the right ventricle. The patient successfully underwent an aortic root replacement in deep hypothermia with re-implantation of the coronary arteries using a modified Bentall procedure and the resection of the corpus alienum. Intraoperative finding reveals 3 greatly adhered gauze compresses, which were most likely forgotten in the operation 34 years ago. PMID- 19121215 TI - Dissociable processes underlying decisions in the Iowa Gambling Task: a new integrative framework. AB - BACKGROUND: The Iowa Gambling Task (IGT) is a common paradigm used to study the interactions between emotions and decision making, yet little consensus exists on the cognitive process determining participants' decisions, what affects them, and how these processes interact with each other. A novel conceptual framework is proposed according to which behavior in the IGT reflects a balance between two dissociable processes; a cognitively demanding process that tracks each option's long-term payoff, and a lower-level, automatic process that is primarily sensitive to loss frequency and magnitude. METHODS: A behavioral experiment was carried out with a modified version of IGT. In this modified version, participants went through an additional phase of interaction, designed to measure performance without further learning, in which no feedback on individual decisions was given. A secondary distractor task was presented in either the first or the second phase of the experiment. Behavioral measures of performance tracking both payoff and frequency sensitivity in choices were collected throughout the experiment. RESULTS: Consistent with our framework, the results confirmed that: (a) the two competing cognitive processes can be dissociated; (b) that learning from decision outcomes requires central cognitive resources to estimate long-term payoff; and (c) that the decision phase itself can be carried out during an interfering task once learning has occurred. CONCLUSION: The experimental results support our novel description of the cognitive processes underlying performance in the Iowa Gambling Task. They also suggest that patients' impairments in this and other gambling paradigms can originate from a number of different causes, including a failure in allocating resources among cognitive strategies. This latter interpretation might be particularly useful in explaining the impairments of patients with ventromedial prefrontal cortex lesions and, by extension, the contribution of this brain region to human decision making. PMID- 19121216 TI - Relationship between body image disturbance and incidence of depression: the SUN prospective cohort. AB - BACKGROUND: Body image disturbance is an increasing problem in Western societies and is associated with a number of mental health outcomes including anorexia, bulimia, body dysmorphia, and depression. The aim of this study was to assess the association between body image disturbance and the incidence of depression. METHODS: This study included 10,286 participants from a dynamic prospective cohort of Spanish university graduates, who were followed-up for a median period of 4.2 years (Seguimiento Universidad de Navarra - the SUN study). The key characteristic of the study is the permanently open recruitment that started in 1999. The baseline questionnaire included information about body mass index (BMI) and the nine figure schemes that were used to assess body size perception. These variables were grouped according to recommended classifications and the difference between BMI and body size perception was considered as a proxy of body image disturbance. A subject was classified as an incident case of depression if he/she was initially free of depression and reported a physician-made diagnosis of depression and/or the use of antidepressant medication in at least one of the follow-up questionnaires. The association between body image disturbance and the incidence of depression was estimated by calculating the multivariable adjusted Odds Ratio (OR) and its 95% Confidence Interval (95% CI), using logistic regression models. RESULTS: The cumulative incidence of depression during follow up in the cohort was 4.8%. Men who underestimated their body size had a high percentage of overweight and obesity (50.1% and 12.6%, respectively), whereas women who overestimated their body size had a high percentage of underweight (87.6%). The underestimation exhibited a negative association with the incidence of depression among women (OR: 0.72, 95% CI: 0.54 - 0.95), but this effect disappeared after adjusting for possible confounding variables. The proportion of participants who correctly perceived their body size was high (53.3%) and gross misperception was seldom found, with most cases selecting only one silhouette below (42.7%) or above (2.6%) their actual BMI. CONCLUSION: We found no association between body image disturbance and subsequent depression in a cohort of university graduates in Spain. PMID- 19121217 TI - Pulmonary Kaposi sarcoma in a human immunodeficiency virus - infected woman: a case report. AB - Approximately 90% to 95% of Kaposi sarcoma cases occur in human immunodeficiency virus - infected homosexual and bisexual men. Pulmonary Kaposi sarcoma is uncommon in women, and rarely considered as a potential cause of diffuse lung disease in women with acquired immunodeficiency syndrome. The disease is usually mistaken clinically for pulmonary infection. A 32-year-old woman was admitted with a 2-month history of dyspnea, evening fever, hemoptysis, weight loss, and generalized adenomegaly. Physical examination showed erythematous macules in the lower limbs. Skin and open lung biopsy demonstrated Kaposi sarcoma. Computerized tomography demonstrated peribronchovascular interstitial thickening. Although uncommon, pulmonary Kaposi sarcoma should be considered in the differential diagnosis of diffuse lung disease in women with AIDS. PMID- 19121218 TI - Mental hospital reform in Asia: the case of Yuli Veterans Hospital, Taiwan. AB - BACKGROUND: Yuli Veterans Hospital (YVH) has been the largest mental hospital for the patients with chronic and severe mental illness in Taiwan for the past 50 years. While this hospital used to be a symbol of hopelessness among patients and their families and an unspoken shame among Taiwan psychiatry and mental health circles it now represents an example of how an old, custodial hospital can be transformed into a very different institution. In this case study we will describe the features of this transformation, which, over the past 20 years, has aimed to help extended stay inpatients with severe mental illness to integrate into the local community of Yuli even though it is not their original home. METHODS: Using historical documents and oral narratives from Yuli inhabitants, workers and patients of YVH, we will offer a case study of the Yuli model. RESULTS: There are four main components of the Yuli model: holistic medical support, vocational rehabilitation, case management, and the residential program. The four components help patients recover two essential features of their lives: vocational life and ordinary daily routines. As the process of recovery evolves, patients gradually regain inner stability, dignity, self-confidence, and a sense of control. The four components are critical to rebuild the structure and order of life of the patients and are indispensable and interdependent parts of one service package. They operate simultaneously to benefit the patients to the greatest degree possible. DISCUSSION: There are many challenges to the further development and financial viability of the model of services developed at YVH. There are also important questions concerning the replicability of the Yuli model in other sociocultural and service system contexts. CONCLUSION: This case study reveals the possibility of transforming a custodial mental hospital into a hospital providing high quality care. Hospital and community are not in opposition. They are part of a continuum of care for the patients. We reinterpret and redefine the boundary and function of hospital and community, and thereby create a new service model, the Yuli Model, to help patients to reintegrate into the community. The Yuli model, which particularly focuses on the needs of people with long-standing illness and prolonged hospital stay, illustrates one approach to linking hospital and community in a creative and constructive manner. PMID- 19121219 TI - Analysis of the platypus genome suggests a transposon origin for mammalian imprinting. AB - BACKGROUND: Genomic imprinting is an epigenetic phenomenon that results in monoallelic gene expression. Many hypotheses have been advanced to explain why genomic imprinting evolved in mammals, but few have examined how it arose. The host defence hypothesis suggests that imprinting evolved from existing mechanisms within the cell that act to silence foreign DNA elements that insert into the genome. However, the changes to the mammalian genome that accompanied the evolution of imprinting have been hard to define due to the absence of large scale genomic resources between all extant classes. The recent release of the platypus genome has provided the first opportunity to perform comparisons between prototherian (monotreme; which appear to lack imprinting) and therian (marsupial and eutherian; which have imprinting) mammals. RESULTS: We compared the distribution of repeat elements known to attract epigenetic silencing across the entire genome from monotremes and therian mammals, particularly focusing on the orthologous imprinted regions. There is a significant accumulation of certain repeat elements within imprinted regions of therian mammals compared to the platypus. CONCLUSIONS: Our analyses show that the platypus has significantly fewer repeats of certain classes in the regions of the genome that have become imprinted in therian mammals. The accumulation of repeats, especially long terminal repeats and DNA elements, in therian imprinted genes and gene clusters is coincident with, and may have been a potential driving force in, the development of mammalian genomic imprinting. These data provide strong support for the host defence hypothesis. PMID- 19121220 TI - Preventing weight gain: the baseline weight related behaviors and delivery of a randomized controlled intervention in community based women. AB - BACKGROUND: Women aged 25-45 years represent a high risk group for weight gain and those with children are at increased risk because of weight gain associated with pregnancy and subsequent lifestyle change. Average self-reported weight gain is approximately 0.60 kg per year, and weight gain is associated with increased risk of chronic disease. There are barriers to reaching, engaging and delivering lifestyle interventions to prevent weight gain in this population. METHODS: This study investigated the baseline weight related behaviors and feasibility of recruiting and delivering a low intensity self-management lifestyle intervention to community based women with children in order to prevent weight gain, compared to standard education. The recruitment and delivery of the cluster-randomized controlled intervention was in conjunction with 12 primary (elementary) schools. Baseline data collection included demographic, anthropometric, behavioral and biological measures. RESULTS: Two hundred and fifty community based women were randomized as clusters to intervention (n = 127) or control (n = 123). Mean age was 40.4 years (SD 4.7) and mean BMI 27.8 kg/m2 (SD 5.6). All components of this intervention were successfully delivered and retention rates were excellent, 97% at 4 months.Nearly all women (90%) reported being dissatisfied with their weight and 72% attempted to self-manage their weight. Women were more confident of changing their diet (mean score 3.2) than physical activity (mean score 2.7). This population perceived they were engaging in prevention behaviors, with 71% reporting actively trying to prevent weight gain, yet they consumed a mean of 68 g fat/day (SD30 g) and 27 g saturated fat/day (SD12 g) representing 32% and 13% of energy respectively. The women had a high rate of dyslipidemia (33%) and engaged in an average of 9187 steps/day (SD 3671). CONCLUSION: Delivery of this low intensity intervention to a broad cross-section of community based women with children is feasible. Women with children are engaging in lifestyle behaviours which do not confer adequate health benefits. They appear to be motivated to attend prevention programs by their interest in weight management. Interventions are required to strengthen and sustain current attempts at achieving healthy lifestyle behaviours in women to prevent weight gain. PMID- 19121221 TI - A first-generation microsatellite-based genetic linkage map of the Siberian jay (Perisoreus infaustus): insights into avian genome evolution. AB - BACKGROUND: Genomic resources for the majority of free-living vertebrates of ecological and evolutionary importance are scarce. Therefore, linkage maps with high-density genome coverage are needed for progress in genomics of wild species. The Siberian jay (Perisoreus infaustus; Corvidae) is a passerine bird which has been subject to lots of research in the areas of ecology and evolutionary biology. Knowledge of its genome structure and organization is required to advance our understanding of the genetic basis of ecologically important traits in this species, as well as to provide insights into avian genome evolution. RESULTS: We describe the first genetic linkage map of Siberian jay constructed using 117 microsatellites and a mapping pedigree of 349 animals representing five families from a natural population breeding in western Finland from the years 1975 to 2006. Markers were resolved into nine autosomal and a Z-chromosome specific linkage group, 10 markers remaining unlinked. The best-position map with the most likely positions of all significantly linked loci had a total sex average size of 862.8 cM, with an average interval distance of 9.69 cM. The female map covered 988.4 cM, whereas the male map covered only 774 cM. The Z chromosome linkage group comprised six markers, three pseudoautosomal and three sex-specific loci, and spanned 10.6 cM in females and 48.9 cM in males. Eighty one of the mapped loci could be ordered on a framework map with odds of >1000:1 covering a total size of 809.6 cM in females and 694.2 cM in males. Significant sex specific distortions towards reduced male recombination rates were revealed in the entire best-position map as well as within two autosomal linkage groups. Comparative mapping between Siberian jay and chicken anchored 22 homologous loci on 6 different linkage groups corresponding to chicken chromosomes Gga1, 2, 3, 4, 5, and Z. Quite a few cases of intra-chromosomal rearrangements within the autosomes and three cases of inter-chromosomal rearrangement between the Siberian jay autosomal linkage groups (LG1, LG2 and LG3) and the chicken sex chromosome GgaZ were observed, suggesting a conserved synteny, but changes in marker order, within autosomes during about 100 million years of avian evolution. CONCLUSION: The constructed linkage map represents a valuable resource for intraspecific genomics of Siberian jay, as well as for avian comparative genomic studies. Apart from providing novel insights into sex-specific recombination rates and patterns, the described maps - from a previously genomically uncharacterized superfamily (Corvidae) of passerine birds - provide new insights into avian genome evolution. In combination with high-resolution data on quantitative trait variability from the study population, they also provide a foundation for QTL-mapping studies. PMID- 19121222 TI - Interactions among type I and type II interferon, tumor necrosis factor, and beta estradiol in the regulation of immune response-related gene expressions in systemic lupus erythematosus. AB - INTRODUCTION: Systemic lupus erythematosus (SLE) is a prototypical autoimmune disease characterized by various clinical manifestations. Several cytokines interact and play pathological roles in SLE, although the etiopathology is still obscure. In the present study we investigated the network of immune response related molecules expressed in the peripheral blood of SLE patients, and the effects of cytokine interactions on the regulation of these molecules. METHODS: Gene expression profiles of peripheral blood from SLE patients and from healthy women were analyzed using DNA microarray analysis. Differentially expressed genes classified into the immune response category were selected and analyzed using bioinformatics tools. Since interactions among TNF, IFNgamma, beta-estradiol (E2), and IFNalpha may regulate the expression of interferon-inducible (IFI) genes, stimulating and co-stimulating experiments were carried out on peripheral blood mononuclear cells followed by analysis using quantitative RT-PCR. RESULTS: Thirty-eight downregulated genes and 68 upregulated genes were identified in the functional category of immune response. Overexpressed IFI genes were confirmed in SLE patient peripheral bloods. Using network-based analysis on these genes, several networks including cytokines--such as TNF and IFNgamma--and E2 were constructed. TNF-regulated genes were dominant in these networks, but in vitro TNF stimulation on peripheral blood mononuclear cells showed no differences in the above gene expressions between SLE and healthy individuals. Co-stimulating with IFNalpha and one of TNF, IFNgamma, or E2 revealed that TNF has repressive effects while IFNgamma essentially has synergistic effects on IFI gene expressions in vitro. E2 showed variable effects on IFI gene expressions among three individuals. CONCLUSIONS: TNF may repress the abnormal regulation by IFNalpha in SLE while IFNgamma may have a synergistic effect. Interactions between IFNalpha and one of TNF, IFNgamma, or E2 appear to be involved in the pathogenesis of SLE. PMID- 19121223 TI - Large scale multiplex PCR improves pathogen detection by DNA microarrays. AB - BACKGROUND: Medium density DNA microchips that carry a collection of probes for a broad spectrum of pathogens, have the potential to be powerful tools for simultaneous species identification, detection of virulence factors and antimicrobial resistance determinants. However, their widespread use in microbiological diagnostics is limited by the problem of low pathogen numbers in clinical specimens revealing relatively low amounts of pathogen DNA. RESULTS: To increase the detection power of a fluorescence-based prototype-microarray designed to identify pathogenic microorganisms involved in sepsis, we propose a large scale multiplex PCR (LSplex PCR) for amplification of several dozens of gene-segments of 9 pathogenic species. This protocol employs a large set of primer pairs, potentially able to amplify 800 different gene segments that correspond to the capture probes spotted on the microarray. The LSplex protocol is shown to selectively amplify only the gene segments corresponding to the specific pathogen present in the analyte. Application of LSplex increases the microarray detection of target templates by a factor of 100 to 1000. CONCLUSION: Our data provide a proof of principle for the improvement of detection of pathogen DNA by microarray hybridization by using LSplex PCR. PMID- 19121224 TI - Influence of immediate and permanent obturators on facial contours: a case series. AB - INTRODUCTION: Rehabilitation of patients after surgical removal of carcinomas in facial skeleton is one of the most difficult therapies of the stomatognathic system. Significant deformation of tissues, dysfunctions of the stomatognathic system with concurrent biological imbalance of the oral cavity environment frequently affect the treatment to become arduous. Scars and contraction of the oral crevice may cause serious psychological deficiencies that are another aspect of the treatment schedule. CASE PRESENTATION: Three Turkish patients ages 46 (male), 61 (male) and 24 (female) who experienced similar operations were rehabilitated with maxillary obturators. The situations was ideal for patient no 1. Patient no 2 could not receive an immediate obturator and patient no 3 rejected using permanent obturator. The paper describes the advantages of a surgical obturator which is constructed before operation and inserted immediately following partial maxillectomy and expresses long term complications when neglecting the use of definite obturator prosthesis, in the light of three cases. CONCLUSION: The primary objective of oral-maxillofacial and plastic surgeons and prosthodontists when treating tumors is to eliminate disease and to improve the quality of life including the facial contours which influences the psychological condition of patient. Neglecting immediate obturator construction may cause serious facial appearance problems due to soft tissue contracture. When permanent obturator is rejected, serious contracture of soft tissues and facial disharmony is inevitable. PMID- 19121225 TI - Failure of E. coli bacteria to induce preterm delivery in the rat. AB - BACKGROUND: We sought to develop a model of bacterially induced preterm delivery in rats to parallel similar models in mice. METHODS: Female Sprague-Dawley rats on day 17 of gestation (normal term = 21-22 days) were inoculated into the uterus with either 2 x 10(9)-7 x 10(10) killed E. coli organisms, 1-4 x 10(8) live E. coli or sterile solution. These inoculations were made either via trans-cervical catheter or by direct intrauterine injection at laparotomy. Animals were then observed for delivery for variable periods up to term. Necropsies were performed and fetal viability was assessed. RESULTS: No rats delivered prematurely after bacterial exposure (27 animals observed for at least 48 hours), and all animals followed to term (n = 3) delivered live pups. No dams exhibited signs of systemic illness. There was a statistically significant but small negative effect of killed E. coli on fetal viability (100% of 80 fetuses from 6 control pregnancies and 93% of 182 fetuses from 14 bacterially-treated pregnancies were alive at necropsy, p = 0.014). Live bacteria had a larger effect on fetal viability, with only 64% of 14 fetuses, 47% of 28 fetuses and 32% of 31 fetuses surviving after trans-cervical administration of 7 x 10(7), 2 x 10(8) and 4 x 10(8) E. coli, respectively. CONCLUSION: Unlike mice, it has proven difficult to induce preterm labor in the rat using E. coli as a stimulating agent. The relevant literature is reviewed and hypotheses are offered to explain this phenomenon. PMID- 19121226 TI - Hepatitis B virus subgenotypes D1 and D3 are prevalent in Pakistan. AB - BACKGROUND: As the hepatitis B genotyping is important for assessing its clinical implications and geographical distribution, the sub-genotypes have been found useful for determination of specific genomic markers related to hepatocarcinogenesis. In Pakistan, there is no reported data on molecular evolutionary analysis of HBV. A study was, therefore, much needed to evaluate the spectra of mutations present in the strains prevalent here. FINDINGS: to confirm specificity of PCR typing, phylogenetic analysis of the pre-S1 region and the divergence was studied through 13 sequences of 362 bp (accession number EF432765 EF432777). A total of 315 serum samples, selected from HBsAg positive patients representing the major ethnic groups, residing in Karachi, Sindh were tested for genotyping. Genotype D (219/315) was found to be the most prevalent (70%) amongst our patients. The rest of the genotypes A and a mixture of A and D (AD) were distributed as 20%, and 10% respectively. Phylogenetic tree demonstrated clustering of 11 samples with subgenotype D1 sequences and the remaining two strains on a branch within D3 samples. All samples intermixed with strains from other countries and were found to be closely related to Indian, Iranian and Egyptian HBV strains with 98.7 - 99.0% homology. CONCLUSION: This study confirms the predominance of genotype D in southeastern Asia and presence of subgenotypes DI and D3 in the Pakistani infected patients. More studies are required to investigate the reason for fewer inclusions of D3 compared to the D1 in Pakistani HBV strains. PMID- 19121227 TI - Correlating the site of tympanic membrane perforation with Hearing loss. AB - BACKGROUND: It is recognized that the size of tympanic membrane perforation is proportional to the magnitude of hearing loss, however, there is no clear consensus on the effect of the location (site) of the perforation on the hearing loss. Hence the study is set to investigate the relationship between the location of perforation on TM and hearing loss. METHODS: A cross-sectional prospective study of consecutive adult patients with perforated TM conducted in the ENT clinic of University College Hospital Ibadan between January 1st 2005 and July 31st 2006. Instruments used for data collection/processing include questionnaires, video and micro-otoscopy, Pure tone audiometer, image J and SPSS packages. RESULTS: Sixty-two patients (22-males, 40-females), aged 16-75 years (mean = 35.4 +/- 4) with 77 perforated ear drums were studied and 15(24.2%) had bilateral TM perforations, 21 (33.9%) right unilateral and 26(41.9%) left unilateral. The locations of the TM perforations were 60(77.9%) central, 6(9.6%) antero-inferior, 4(5.2%) postero-inferior, 4(5.2%) antero-superior and 3(3.9%) postero-superior respectively with sizes ranging from 1.51%-89.05%, and corresponding hearing levels 30 dB - 80 dB (59% conductive and 41% mixed). Fifty nine percent had pure conductive hearing loss and the rest mixed. Hearing losses (dBHL) increased with the size of perforations (P = 0.01, r = 0.05). Correlation of location of perforations with magnitude of hearing loss in acute TM perorations was (P = 0.244, r = 0.273) and for chronic perforations (p = 0.047 & r = 0.31). CONCLUSION: The location of perforation on the tympanic membrane (TM) has no effect on the magnitude of hearing loss in acute TM perforations while it is significant in chronic ones. PMID- 19121228 TI - Redox biology of Mycobacterium tuberculosis H37Rv: protein-protein interaction between GlgB and WhiB1 involves exchange of thiol-disulfide. AB - BACKGROUND: Mycobacterium tuberculosis, an intracellular pathogen encounters redox stress throughout its life inside the host. In order to protect itself from the redox onslaughts of host immune system, M. tuberculosis appears to have developed accessory thioredoxin-like proteins which are represented by ORFs encoding WhiB-like proteins. We have earlier reported that WhiB1/Rv3219 is a thioredoxin like protein of M. tuberculosis and functions as a protein disulfide reductase. Generally thioredoxins have many substrate proteins. The current study aims to identify the substrate protein(s) of M. tuberculosis WhiB1. RESULTS: Using yeast two-hybrid screen, we identified alpha (1,4)-glucan branching enzyme (GlgB) of M. tuberculosis as a interaction partner of WhiB1. In vitro GST pull down assay confirmed the direct physical interaction between GlgB and WhiB1. Both mass spectrometry data of tryptic digests and in vitro labeling of cysteine residues with 4-acetamido-4' maleimidyl-stilbene-2, 2'-disulfonic acid showed that in GlgB, C95 and C658 are free but C193 and C617 form an intra-molecular disulfide bond. WhiB1 has a C37XXC40 motif thus a C40S mutation renders C37 to exist as a free thiol to form a hetero-disulfide bond with the cysteine residue of substrate protein. A disulfide mediated binary complex formation between GlgB and WhiB1C40S was shown by both in-solution protein-protein interaction and thioredoxin affinity chromatography. Finally, transfer of reducing equivalent from WhiB1 to GlgB disulfide was confirmed by 4-acetamido-4' maleimidyl-stilbene 2, 2'-disulfonic acid trapping by the reduced disulfide of GlgB. Two different thioredoxins, TrxB/Rv1471 and TrxC/Rv3914 of M. tuberculosis could not perform this reaction suggesting that the reduction of GlgB by WhiB1 is specific. CONCLUSION: We conclude that M. tuberculosis GlgB has one intra-molecular disulfide bond which is formed between C193 and C617. WhiB1, a thioredoxin like protein interacts with GlgB and transfers its electrons to the disulfide thus reduces the intra-molecular disulfide bond of GlgB. For the first time, we report that GlgB is one of the in vivo substrate of M. tuberculosis WhiB1. PMID- 19121229 TI - Anti-atopic dermatitis effects and the mechanism of lactic acid bacteria isolated from Mongolian fermented milk. AB - We investigated the anti-allergic effects of one strain (T120) of a lactic acid bacteria (LAB) isolated from Mongolian fermented milk using atopic dermatitis (AD) model mice (NC/Nga mice). Strain T120 has already been identified as Enterococcus faecium and shown to induce strong production of IL-12 (Kimura et al. 2006). In in vitro studies, strain T120 suppressed total IgE production and induced IL-12 and IFN-gamma production by splenocytes of NC/Nga mice. The additional examination of various neutralization antibodies was performed to elucidate in detail the mechanism of depressed IgE production by strain T120. As a result, it became clear that IL-12 induced by strain T120 increased production of IFN-gamma and total IgE production was mainly controlled by the IFN-gamma. In order to define the cells which produce IL-12 powerfully by this strain, antigen presenting cells (APCs) such as macrophages and dendritic cells (DCs) were removed from the splenocytes, and the reactivity of these cells to the strain was examined. Induction of IL-12 and IFN-gamma by strain T120 became significantly very low by removal of APCs from splenocytes. Therefore, it was clear that strain T120 acted on APCs and induced production of IL-12. Further, this strain enhanced the production of IL-10 by splenocytes. In in vivo studies, intraperitoneal injection of strain T120 inhibited serum IgE elevation and atopic dermatitis symptoms in NC/Nga mice. These results suggest that an anti-allergic effect of strain T120 depends on the increased production of IL-12 by APCs activated by the strain and following the increased production of IFN-gamma. Further, activation of regulatory T cells by strain T120 may inhibit atopic disease. PMID- 19121230 TI - Synbiotic effect of Lactobacillus helveticus M92 and prebiotics on the intestinal microflora and immune system of mice. AB - The synbiotic effect of the oral treatment of Swiss albino mice with milk-based diets supplemented with Lactobacillus helveticus M92 and various kinds of prebiotics was investigated. Survival, competition, adhesion and colonization, as well as, immunomodulating capability of Lb. helveticus M92, in synbiotic combination, in the gastrointestinal tract (GIT) of mice, were monitored. After the mice were fed with synbiotics, the lactic acid bacteria (LAB) counts in faeces were increased and reduction of enterobacteria and sulphite-reducing clostridia was observed. Similar results were obtained in homogenates of small and large intestine of mice on the 1st and 14th day, after feeding with synbiotics. After the mice were orally given viable Lb. helveticus M92 cells, alone or in combination with prebiotic, the concentration of faecal SIgA and total serum IgA antibodies from all immunized mice were higher compared with the control. The specific humoral immune response was not evoked after oral administration, therefore their synbiotic application is suitable. Among inulin, lactulose and raffinose, Lb. helveticus M92 in combination with inulin, has shown the best synbiotic effect on intestinal and faecal microflora and immune system of mice. PMID- 19121231 TI - Interaction between Lactobacillus kefir and Saccharomyces lipolytica isolated from kefir grains: evidence for lectin-like activity of bacterial surface proteins. AB - Several microbial interactions involving yeast and lactobacilli have been suggested in fermented products. Co-aggregation between Lactobacillus kefir and yeast Saccharomyces lipolytica isolated from kefir grains was studied by scanning electron microscopy and aggregation assays. Six out of twenty Lb. kefir strains were able to co-aggregate with Sacch. lipolytica CIDCA 812 and showed thermolabile non-covalently bound surface molecules involved in this interaction. Co-aggregation inhibition after Lb. kefir pre-treatment with 5 m-LiCl or 20 g SDS/l showed that bacterial S-layer proteins play an important role in this interaction. Presence of different sugar (mannose, sucrose and fructose) or yeast pre-treatment with sodium periodate inhibited co-aggregation between Lb. kefir and Sacch. lipolytica. Co-aggregating Lb. kefir strains were also able to agglutinate with human red blood cells and they lost this ability after treatment with 5 m-LiCl. These results and the capacity of purified S-layer proteins of Lb. kefir to haemagglutinate, strongly suggest that a lectin-like activity of bacterial surface proteins (S-layer) mediates the aggregation with yeast cells. PMID- 19121232 TI - Evaluation of the serum fructosamine test to monitor plasma glucose concentration in the transition dairy cow. AB - The usefulness of the serum fructosamine (Fser) to monitor the retrospective glucose concentrations in transitional dairy cows (n=17) was evaluated. In weekly blood samples (3 weeks before to 5 weeks after calving) concentrations of plasma glucose and serum fructosamine, beta-hydroxybutyrate (beta OHB) and total proteins were determined. The observed Fser concentrations (271+/-55 mean value, range 152-423 mumol/l) were within the range reported in the literature, and showed a progressive and significant decrease after calving. Mean plasma glucose concentration was 60.6+/-5.0 (range 39.9-82.2) mg/dl increasing from week 3 before calving to the week of calving and then decreasing during the next 5 weeks of lactation. This decrease was coincident with inverse relationships between plasma glucose and milk yield (P=0.03) and serum beta OHB (P<0.001). Linear regression analysis performed between serum fructosamine and (a) plasma glucose concentration of the same sampling and (b) plasma glucose concentration of 1, 2 and 3 weeks preceding the sampling, did not show significant and systematizing positive correlations. Persistent hypoproteinaemias that could affect the fructosamine concentrations were not found: mean value and range of serum proteins was 6.3+/-1.0 and 4.8-7.8 g/dl, respectively, and no correlation was found between serum proteins and Fser (P=0.26). Results did not support the possibility of retrospective monitoring of the plasma glucose concentration by serum fructosamine in dairy cows in the transition period. PMID- 19121233 TI - Prevalence of intramammary infection in Dutch dairy herds. AB - A survey was carried out in 2003 in 49 dairy herds to determine the overall and pathogen-specific prevalence of intramammary infection (IMI) in Dutch dairy herds, and to compare the distribution with four studies performed from 1973 to 1985 in The Netherlands. Herds were randomly selected stratified over the 12 Dutch provinces, had at least 40 lactating cows and participated in the Dutch milk recording system. Quarter milk samples were collected from all 408 cows with a somatic cell count (SCC) >or=250,000 cells/ml and 145 heifers with SCC >or=150,000 cells/ml at the last milk test before the farm visit. Additionally, samples were collected from 519 (approximately 25%) of the remaining low-SCC cows and heifers with a SCC at the last milk test before the farm visit of <250 000 and <150 000 cells/ml, respectively. Bacterial growth occurred in 37.3% of milk samples of high-SCC cows and in 21.1% of low-SCC cows. Coagulase-negative staphylococci (CNS) were the most frequently isolated group of bacteria (10.8% of quarters) and were found in all herds. Prevalence of Staphylococcus aureus IMI was lower in 2003 than in 1973, respectively 1.8% and 6.2% of quarters. Prevalence of Streptococcus uberis and Str. dysgalactiae IMI was almost the same in the five samplings during the 30-year period, at 1.1-1.7 and 0.9-1.5%, respectively. Str. agalactiae was not found in this study. Prevalence of CNS IMI was higher in lactating heifers, while prevalence of Str. uberis, Str. dysgalactiae and penicillin-resistant Staph. aureus IMI was higher in older cows. Because distribution of pathogens changes over time, herd-level samples for bacteriological culturing must be taken regularly to monitor udder health. Additionally, national mastitis prevalence studies give important information through monitoring the national udder health status. PMID- 19121234 TI - Effect of bovine lactoferrin on the internalization of coagulase-negative staphylococci into bovine mammary epithelial cells under in-vitro conditions. AB - Coagulase-negative staphylococci (CNS) have emerged as bovine mastitis pathogens in many countries. CNS mastitis is generally mild but can persist in the udder for long periods. Pathogenesis of CNS intramammary infection is not well understood. In the present study, adhesion, invasion and intracellular replication of twenty-two CNS strains isolated from bovine mastitis and the effect of bovine lactoferrin (bLf) on the internalization were studied in vitro in a bovine mammary epithelial (BME) cell model. The CNS strains were of Staphylococcus chromogenes, Staph. simulans, Staph. epidermidis, Staph. haemolyticus and Staph. cohnii urealyticus; two strains of Staph. aureus were used as controls. Seven of the CNS strains originated from persistent and five from transient mastitis infections. The in-vitro susceptibility of the strains to bLf was also investigated. All CNS species examined had an adhesive ability equal to that of Staph. aureus, but internalization varied among staphylococcal strains. The antagonistic effect of bLf on the adhesion and invasion of CNS strains was weak, but bLf significantly decreased intracellular replication and replication rates of CNS. No correlation between the in-vitro susceptibility of the strain to bLf or internalization among clinical signs of mastitis was established. No difference between the persistent and transient CNS strains in adhesion, invasion or replication rate was recorded. This in-vitro BME cell model can be used to study the virulence potential of mastitis pathogens, although the severity and persistence of eventual infections shall be further investigated in vivo. The role of bLf in intramammary infection caused by CNS may be limited. PMID- 19121235 TI - Prevalence of pathogens in milk samples of dairy cows with clinical mastitis and in heifers at first parturition. AB - Prevalence of mastitis pathogens in milk samples from dairy cows and heifers was studied over a period of 1 year (Aug 2005-Aug 2006) in ten dairy herds in Germany. Milk samples (n=8240) were collected from heifers without clinical mastitis at parturition (n=6915), from primiparous cows with clinical mastitis (n=751) and from older cows with clinical mastitis (n=574). Coagulase negative staphylococci (CNS) were the predominant group of bacteria isolated (46.8% of samples) from clinically healthy quarters of primiparous cows around parturition, followed by streptococci (12.6%), coliforms (4.7%) and Staphylococcus aureus (4.0%). Thirty-three percent of samples were negative on culture (range on farm level, 12.0-46.4%). In cases of clinical mastitis in primiparous and older cows, streptococci were the predominant finding (32.1 and 39.2%) followed by CNS (27.4 and 16.4%), coliforms (10.3 and 13.1%) and Staph. aureus (10.0 and 11.7%). Negative results were obtained from 21.3% (range, 0.0-30.6%) and 19.5% (range, 0.0-32.6%) of these samples. Results indicated substantial differences in the prevalence of pathogens among herds. There was a positive within-herd correlation between the monthly prevalences for Streptococcus dysgalactiae between the three groups of samples. This correlation was also found between clinical samples of primiparous and older cows for Staph. aureus. These correlations were not found for the other pathogens. Besides herd, prevalence of pathogens was influenced by parity, type of sample and season. PMID- 19121236 TI - Milk production and composition in Malaguena dairy goats. Effect of genotype for synthesis of alpha s1-casein on milk production and its interaction with dietary protein content. AB - A study was carried out to analyse the production and composition of milk from Malaguena goats of different genotypes for the content of alpha s1-casein. Twenty goats were used, half of them with a high (HG) and half with a low (LG) genetic capability for alpha s1-casein synthesis. Among the ten HG goats, seven had BB and three had AB genotypes, whereas there were seven EF and three FF genotypes among the LG goats. The goats were fed a diet differing in crude protein content, 136 and 177 g/kg dry matter for diet 1 (D1) and diet 2 (D2), respectively. For each genotype group, a trial was conducted, with each of the two diets in a two period balanced change-over design, in which the milk production and composition were analysed. The genotype was found to determine milk composition, with that produced by the HG goats being higher in total solids, protein, alpha s1-casein, alpha s2-casein and fat. Analysis of milk production values and, especially, those of the yields of the different components, revealed an interaction between the factors. HG goats that consumed D1 were more productive than LG goats, but this ceased to be so when the D1 diet was replaced by D2. However, LG goats, although less productive than HG ones when the D1 diet was consumed, showed increased productive capacity when D1 was replaced by D2. The main difference for the fatty acid profile of the dairy fat is that the milk fat produced by the HG goats contained higher levels of short and medium-chain fatty acids. From these results, it is concluded that, in terms of favouring production, the advantage of replacing animals with a low capacity for alpha s1-casein synthesis with those of a high capacity seems to depend on the protein level in the diet. Similarly, the advantage of changing to a diet of higher protein content depended on the animal's genotype. PMID- 19121237 TI - Supplemental fish oil does not alter immune competence or the pathophysiological response to an intramammary infusion of endotoxin in peri-partum multiparous Holstein cows. AB - The objective was to determine the effects of supplementing the diet with fish oil during the peri-partum period on the immune competence and the pathophysiological response to a lipopolysaccharide-induced mastitis challenge. Multiparous Holstein cows (n=30) were completely randomized to one of two treatments at 3 weeks pre-partum. Treatments differed only in the source of supplemental lipid and included either Energy Booster or fish oil. Treatment diets were fed from -21 d relative to expected date of parturition until 10 d post partum. Treatments were fed as a bolus prior to the a.m. feeding. The dose of lipid during the pre-partum period was 250 g/d, whereas the amount of lipid supplemented post partum was adjusted to the level of intake, approximately 0.92% of the previous day's dry matter intake. Ex-vivo analyses of immune competence were measured including the antimicrobial activity of whole blood against Escherichia coli, Salmonella typhimurium and Candida albicans as well as the production of interferon-gamma by peripheral blood mononuclear cultures. At 7 days in milk cows were infused with 100 microg of Esch. coli lipopolysaccharide into one rear quarter. Supplementing fish oil increased plasma concentrations of eicosapentaenoic and docosahexaenoic acids, but had no affect on the proportions of arachidonic acid at calving. Fish oil did not influence the production of interferon-gamma or the antimicrobial activity of whole blood against any of the microorganisms. Furthermore, fish oil had no ameliorative effect on either the local or the systemic acute phase response following an intramammary lipopolysaccharide challenge in early lactating Holstein cows. Supplementing fish oil in the diet of peri-partum cows will not protect them from deleterious effects of an excessive acute phase response. PMID- 19121238 TI - Monitoring major mastitis pathogens at the population level based on examination of bulk tank milk samples. AB - The objective was to determine the proportions and bacterial counts of major mastitis pathogens in samples of bulk tank milk (BTM), as well as to clarify the relationship between these bacteria counts and bulk tank somatic cell count (BTSCC). The purpose was to judge the importance of the counts of mammary pathogens for BTSCC at the population level. Samples of BTM were collected from 268 randomly selected anonymous dairy herds (with approximately 29 000 dairy cows). Staphylococcus aureus, other coagulase-positive staphylococci, Streptococcus agalactiae and Streptococcus dysgalactiae were grouped as contagious pathogens, and Streptococcus uberis, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis and Enterococcus faecium as environmental pathogens. Based on assessment of the dominant pathogen in each herd, environmental pathogens were found to dominate. Counts of specific mammary pathogens in BTM samples did not exceed a geometric mean of 1000 cfu/ml. Significant differences were documented in BTSCC between BTM samples containing Str. agalactiae, Staph. aureus, and Str. dysgalactiae and BTM samples that were pathogen-free. Geometric means of BTSCC associated with these pathogens were noticeably higher than the overall BTSCC (Str. agalactiae 243 628; Staph. aureus 205 610; Str. dysgalactiae 203 978; overall 173 000 cells/ml). It follows that the somatic cell count (SCC) associated with these pathogens contributed substantially to the overall BTSCC. Environmental pathogens predominated in samples of BTM with SCC <300x103/ml and contagious mastitis pathogens predominated in BTM samples with SCC >300x103/ml. No correlation was detected between bacterial counts of specific pathogens and BTSCC. This study revealed that the assessment of bacterial counts of mammary pathogens in samples of BTM in relation to BTSCC is applicable for the monitoring of changes in the occurrence of major mastitis pathogens in dairy herds at a national level. PMID- 19121239 TI - Suitability of high pressure-homogenized milk for the production of probiotic fermented milk containing Lactobacillus paracasei and Lactobacillus acidophilus. AB - High pressure homogenization (HPH) is one of the most promising alternatives to traditional thermal treatment for food preservation and diversification. In order to evaluate its potential for the production of fermented milks carrying probiotic bacteria, four types of fermented milks were manufactured from HPH treated and heat treated (HT) milk with and without added probiotics. Microbiological, physicochemical and organoleptic analyses were carried out during the refrigerated period (35 d at 4 degrees C). HPH application to milk did not modify the viability of the probiotic cultures but did increase the cell loads of the starter cultures (ca. 1 log order) compared with traditional products. The coagula from HPH-milk was significantly more compacted (P<0.05) (higher firmness) than that obtained with HT-milk, and it had the highest values of consistency, cohesiveness and viscosity indexes compared with fermented milks produced without HPH treatment. All the samples received high sensory analysis scores for each descriptor considered. HPH treatment of milk can potentially diversify the market for probiotic fermented milks, especially in terms of texture parameters. PMID- 19121240 TI - Use of FTIR and mass spectrometry for characterization of glycated caseins. AB - This study investigates the potential of Fourier transform infrared spectroscopy (FTIR) to monitor glycation-induced changes in protein structure. Aqueous solutions of sodium caseinate and glucose (1:2 w/w, pH 6.7) were heated at 90 degrees C for 0, 10, 20, 40 and 60 min. Evidence for caseinate glycation was obtained by mass spectrometry techniques (electrospray (ESI) and matrix-assisted laser desorption ionisation (MALDI)). FTIR was able to discriminate between glycated and non-glycated sodium caseinate, when the data were analysed by multivariate statistical methods; principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA). The techniques used were complementary and provided different levels of information about the glycated samples. PMID- 19121241 TI - Focusing on casein gene cluster and protein profile in Garganica goat milk. AB - A survey was carried out in eight goat dairy farms, a total of 71 individual Garganica goat milk samples were collected for genomic DNA extraction. Casein alleles and haplotype frequencies of Garganica population were estimated. Individual milks were also analysed for chemical composition, rheological properties, and protein profile. The strong A* allele of CSN1S1 was predominant in the population investigated, the weak allele F of CSN1S1 showed a relatively high frequency and the null alleles N and 01 were first observed in this breed. At CSN1S2 locus the strong A* allele was the most frequent, followed by the F allele and the null allele. The strong A* allele was predominant at CSN2 locus, and relatively high incidence of null allele 0 was observed. CSN3 locus was monomorphic for B* allele. The exact test of sample differentiation based on haplotype frequencies discriminate the farms into two groups characterized by the highest frequency of strong (S-CSN1S1) or weak (W-CSN1S1) alleles at CSN1S1. Protein and casein contents were higher in the group characterized by strong allele than in the group with weak allele at CSN1S1. The 2D electrophoresis technique was performed to screen goat casein variability at the protein level and to evaluate global casein genotype (alphas1, alphas2, beta and kappa-CN). Gels displayed the protein profile associated with casein genotype, and demonstrated differences in the protein expression deriving from interactions between loci. The variability of goat casein loci in Garganica goat breed could be exploited to differentiate the population on the basis of milk utilization and could represent a strategy to preserve the genotype of this autochthonous breed. PMID- 19121242 TI - Exopolysaccharide-producing mesophilic lactic cultures for preparation of fat free Dahi - an Indian fermented milk. AB - Forty seven exopolysaccharide (EPS) producing mesophilic lactic acid bacteria have been isolated from Dahi and raw milk and selected cultures were evaluated for their influence on rheological and sensory properties of fat-free Dahi. Two isolates namely B-6 and KT-24 that showed promising technological attributes were identified as Lc. lactis subsp. lactis strains. B-6 produced 184+/-2 mg/l EPS in deproteinized whey medium compared with 193+/-1 mg/l by KT-24. EPS produced by B 6 was a heteropolysaccharide (consisting of glucose and mannose, 1:7 x 4) with molecular weight of 3.0x104 Da whereas KT-24 EPS was a homopolysaccharide (rhamnose) having molecular weight of 4.5x104 Da. Both EPS producing cultures showed significant changes in rheological and sensory properties of fat-free Dahi. Dahi prepared by these cultures was more viscous, adhesive, sticky, showed lower susceptibility to whey separation, and received higher sensory scores than Dahi prepared with non-EPS producing culture. PMID- 19121243 TI - Crude goat whey fermentation by Kluyveromyces marxianus and Lactobacillus rhamnosus: contribution to proteolysis and ACE inhibitory activity. AB - Unsupplemented acid goat whey containing 0.96% protein and 2.76% lactose was fermented aerobically with 32 microflora extracted from various raw milk cheeses and dairy products. These microflora were screened for their ability to hydrolyse whey proteins (alpha-lactalbumin and/or beta-lactoglobulin) and to generate peptides inhibitors of Angiotensin I Converting Enzyme. Five microflora were able to degrade whey protein. The most efficient microflora was able to fully hydrolyse alpha-lactalbumin and to a lesser extend beta-lactoglobulin. It was extracted from Bamalou des Pyrenees cheese. Micro-organisms involved consisted of yeast Kluyveromyces marxianus and lactobacillus Lactobacillus rhamnosus. Both were able to produce ACE inhibitory peptides after whey fermentation. PMID- 19121245 TI - Camera in the emergency department: the evolution of stroke telemedicine. PMID- 19121246 TI - Acute pericarditis: appendicitis of the heart? PMID- 19121244 TI - Stroke telemedicine. AB - Stroke telemedicine is a consultative modality that facilitates care of patients with acute stroke at underserviced hospitals by specialists at stroke centers. The design and implementation of a hub-and-spoke telestroke network are complex. This review describes the technology that makes stroke telemedicine possible, the members that should be included in a telestroke team, the hub-and-spoke characteristics of a telestroke network, and the format of a typical consultation. Common obstacles to the practice of telestroke medicine are explored, such as medicolegal, economic, and market issues. An example of a state based telestroke network is thoroughly described, and established international telestroke networks are presented and compared. The opportunities for future advances in telestroke practice, research, and education are considered. PMID- 19121247 TI - The business of providing high-quality content to Mayo Clinic Proceedings' readers: 2009 and beyond. PMID- 19121248 TI - Frequency and predictors of urgent coronary angiography in patients with acute pericarditis. AB - OBJECTIVES: To determine the frequency of urgent coronary angiography in patients with acute pericarditis and to examine clinical characteristics associated with coronary angiography. PATIENTS AND METHODS: This is a retrospective analysis of all incident cases of acute viral or idiopathic pericarditis evaluated at Mayo Clinic's site in Rochester, MN, between January 1, 2000, and December 31, 2006. The main outcome measures were use of urgent coronary angiography and rate of concomitant coronary artery disease in patients with pericarditis. RESULTS: There were 238 patients with a final diagnosis of acute pericarditis (mean age, 47.7+/ 17.9 years; 157 [66.0%] were male). On the initial electrocardiogram, 146 patients (61.3%) had ST-segment elevation, and 92 (38.7%) had no ST-segment elevation. Coronary angiography was performed in 40 patients (16.8% of all patients); the frequency was 5-fold higher among those with ST-segment elevation (24.7% vs 4.3%; P<.001). Additionally, 7 patients (4.8%) with ST-segment elevation received thrombolytics before transfer to our institution; no patients without ST-segment elevation received thrombolysis (P=.05). Characteristics associated with a higher likelihood of coronary angiography included typical anginal chest pain, ST-segment elevation, previous percutaneous coronary intervention, elevated troponin T values, diaphoresis, and male sex. Coronary angiography revealed concomitant mild to moderate coronary artery disease in 14 (35.0%) of the 40 patients who underwent this procedure. CONCLUSION: Urgent coronary angiography is commonly performed in patients with acute pericarditis, particularly those with ST-segment elevation, typical myocardial infarction symptoms, and elevated troponin T values. Coronary artery disease was present angiographically in one-third of patients undergoing the procedure. Although patients with ST-segment elevation myocardial infarction must receive prompt reperfusion, clinicians must also consider the diagnosis of pericarditis to avoid unneeded coronary angiography. PMID- 19121249 TI - Incidence and temporal trends of primary immunodeficiency: a population-based cohort study. AB - OBJECTIVE: To determine the incidence and temporal trends of primary immunodeficiency diseases (PIDs) and examine whether an association exists between delayed diagnosis and increased morbidity. PATIENTS AND METHODS: We performed a historical cohort study to describe the epidemiology of PIDs in Olmsted County, Minnesota, during a 31-year period from January 1, 1976, through December 31, 2006, using the Rochester Epidemiology Project. Incidence and trends over time, presence of comorbid conditions, and trends in management were determined. RESULTS: During the 31-year study period, 158 new cases of PIDs were diagnosed, with an overall incidence rate of 4.6 per 100,000 person-years. The rate of PIDs from 2001 through 2006 (10.3 per 100,000 person-years) was nearly 5 times higher than that from 1976 through 1980 (2.4 per 100,000 person-years). The associations between continuous variable(s) and categorical outcome(s) were assessed by using the Wilcoxon rank sum test. Longer delay in diagnosis was significantly associated with recurrent sinusitis (P<.001), recurrent pneumonia (P=.03), and subsequent treatment with immunoglobulins (P<.001). On the basis of Kaplan-Meier survival estimates, the proportion of patients surviving at 10 years after diagnosis was 93.5% (95% confidence interval, 85.9%-97.1%). However, older age at diagnosis was significantly associated with mortality (P=.01). CONCLUSION: This is one of the first population-based studies to examine the temporal trends of PIDs. The incidence of PIDs increased markedly between 1976 and 2006. In this cohort, a delay in diagnosis was common and was associated with increased morbidity. Despite substantial morbidity, most patients with PIDs can expect a normal life span. PMID- 19121250 TI - Congenital heart disease associated with selective serotonin reuptake inhibitor use during pregnancy. AB - OBJECTIVE: To determine the risk of congenital cardiac abnormalities associated with use of selective serotonin reuptake inhibitors (SSRIs) during pregnancy. PATIENTS AND METHODS: We conducted a retrospective review of the medical records of all pregnant women presenting at Mayo Clinic's site in Rochester, MN, from January 1, 1993, to July 15, 2005, and identified 25,214 deliveries. A total of 808 mothers were treated with SSRIs at some point during their pregnancy. We reviewed the medical records of the newborns exposed to SSRIs during pregnancy to analyze their outcomes, specifically for congenital heart disease and persistent pulmonary hypertension of the newborn. RESULTS: Of the study patients, 808 (3.2%) took an SSRI at some point during the antenatal period. Of the 25,214 deliveries, 208 newborns (0.8%) were diagnosed as having congenital heart disease. Of the 808 women exposed to SSRI during pregnancy, 3 (0.4%) had congenital heart disease compared with 205 (0.8%) of the 24,406 women not exposed to an SSRI (P=.23). Of the total number of deliveries, 16 newborns were diagnosed as having persistent pulmonary hypertension of the newborn, none of whom had exposure to SSRIs (P>.99). CONCLUSION: Our data are reassuring regarding the safety of using SSRIs during pregnancy. PMID- 19121251 TI - Effect of second-generation sulfonylureas on survival in patients with diabetes mellitus after myocardial infarction. AB - OBJECTIVE: To examine possible adverse effects of sulfonylureas on survival among patients with diabetes mellitus (DM) who experience a myocardial infarction (MI). PATIENTS AND METHODS: Residents of Olmsted County, Minnesota, with an MI that met standardized criteria from January 1, 1985, through December 31, 2002, were followed up for mortality. RESULTS: Among 2189 patients with MI (mean+/-SD age, 68+/-14 years; 1237 men [57%]), 409 (19%) had DM. The 23 patients treated with first-generation sulfonylureas, biguanides, or thiazolidinediones were excluded from analyses. Among the remaining 386 patients with DM, 120 (31%) were taking second-generation sulfonylureas, 180 (47%) were taking insulin, and 86 (22%) were receiving nonpharmacological treatment. Patients with DM treated with second generation sulfonylureas were more likely to be men and have higher creatinine clearance than those treated with insulin. After adjusting for age, sex, Killip class, duration of DM, creatinine clearance, and reperfusion therapy or revascularization, patients treated with second-generation sulfonylureas had a lower risk of death than did diabetic patients receiving insulin (hazard ratio, 0.41; 95% confidence interval, 0.21-0.80; P=.009). CONCLUSION: These population based data do not support the concern about an adverse effect of second generation sulfonylureas on survival after MI and underscore the importance of population-based studies of surveillance of drug safety. PMID- 19121252 TI - Reasons to exercise caution when considering a screening program for type 2 diabetes mellitus. PMID- 19121253 TI - Back to Wilson and Jungner: 10 good reasons to screen for type 2 diabetes mellitus. PMID- 19121255 TI - Subclinical hypothyroidism: an update for primary care physicians. AB - Subclinical hypothyroidism (SCH), also called mild thyroid failure, is diagnosed when peripheral thyroid hormone levels are within normal reference laboratory range but serum thyroid-stimulating hormone (TSH) levels are mildly elevated. This condition occurs in 3% to 8% of the general population. It is more common in women than men, and its prevalence increases with age. Of patients with SCH, 80% have a serum TSH of less than 10 mIU/L. The most important implication of SCH is high likelihood of progression to clinical hypothyroidism. The possibility that it is a cardiovascular risk factor has been a subject of debate. Large-scale randomized studies are needed for evidence-based recommendations regarding screening for mild thyroid failure and levothyroxine therapy for this condition. Currently, the practical approach is routine levothyroxine therapy for persons with a persistent serum TSH of more than 10.0 mIU/L and individualized therapy for those with a TSH of less than 10.0 mIU/L. PMID- 19121254 TI - Secondary prevention of atherothrombotic events after ischemic stroke. AB - Atherosclerotic vascular disease is the leading cause of ischemic stroke, resulting in occlusive or severely stenotic lesions of major intracranial or extracranial arteries and narrowing of small penetrating arteries of the brain. Atherosclerosis of the coronary arteries (ie, coronary artery disease) is an indirect cause of cardioembolic stroke secondary to myocardial infarction. Ischemic heart disease may also be complicated by atrial fibrillation and cardioembolic stroke. Prevention of recurrent stroke and other ischemic events, including myocardial infarction, is a key component of treatment for patients with symptomatic ischemic cerebrovascular disease. Prevention of recurrent stroke involves controlling those factors that promote the course of atherosclerosis, including hypertension, hyperlipidemia, diabetes mellitus, and smoking, as well as such local interventions as carotid endarterectomy and endovascular treatment. Nevertheless, administration of antiplatelet agents remains the core of management for preventing recurrent stroke and other cardiovascular events in at risk patients. PMID- 19121256 TI - 25-year-old man with flank pain, hematuria, and proteinuria. PMID- 19121257 TI - Cannabinoid hyperemesis relieved by compulsive bathing. AB - Cannabinoid hyperemesis is a clinical syndrome characterized by repeated vomiting and associated learned compulsive hot water bathing behavior due to long-term marijuana use. Research has identified type 1 cannabinoid receptors in the intestinal nerve plexus that have an inhibitory effect on gastrointestinal motility. This inhibitory effect may lead to hyperemesis in marijuana users. The thermoregulatory role of endocannabinoids may be responsible for the patient's need to take hot showers. We report 2 cases of cannabinoid hyperemesis that demonstrate this unusual adverse effect of marijuana use. PMID- 19121259 TI - Delirium and pulmonary embolism in the elderly. PMID- 19121258 TI - Perioperative cardiovascular assessment of patients undergoing noncardiac surgery. AB - Preoperative assessment of the cardiac patient before noncardiac surgery is common in the clinical practice of the medical consultant, anesthesiologist, and surgeon. Currently, most noncardiac surgical procedures are performed for patients of advanced age, and the number of such surgeries is likely to increase with the aging of the population. These same patients have an increased prevalence of cardiovascular disease, especially ischemic heart disease, which is the primary cause of perioperative morbidity and mortality associated with noncardiac surgery. Since 1996, 3 American College of Cardiology/American Heart Association guideline documents have been published, each reflecting the available literature, with recommendations for the preoperative cardiovascular evaluation and treatment of the patient undergoing noncardiac surgery. Our review describes the 2007 American College of Cardiology/American Heart Association guidelines, the most recent revision, focusing on a newly recommended 5-step algorithmic approach to managing this clinical problem, particularly for the patient with known or suspected coronary heart disease. Continued emphasis should be given to preoperative clinical risk stratification, with noninvasive testing reserved for those patients in whom a substantial change in medical management would be anticipated based on results of testing. Pharmacologic therapy holds more promise than coronary revascularization for the reduction of major adverse perioperative cardiac events that might complicate noncardiac surgery. PMID- 19121260 TI - Midventricular variant of transient apical ballooning: a likely demonstration of its pathophysiologic mechanism. PMID- 19121261 TI - [For the service of internal medicine]. PMID- 19121262 TI - [Clinicopathological sessions in the XXI century]. PMID- 19121263 TI - [Evolution of opiate use in two Primary Health Care Areas in the community of Madrid (Spain) and their referral hospitals between 2002 and 2006]. AB - BACKGROUND AND OBJECTIVE: Describe the evolution of prescription opioids from 2002 to 2006 in Primary Health Care Area 2 and 5 in the Madrid Community and its referral hospitals (University Hospitals of la Princesa and La Paz). MATERIALS AND METHODS: A retrospective observational study of prescriptions made by calculating defined daily doses per million inhabitants. RESULTS: In Primary Care, transdermal fentanyl (78.39% of the total in 2006 in area 2 and 76.69% in the area 5) is the opioid prescribed most on the third step on the WHO analgesic ladder. This is followed by buprenofin (12.30% and 13.23%) and morphine (6.94% and 8.45%). As for the second step, tramadol is the most prescribed opioid with 85.12% and 83.56% of total consumption. We see a growth in consumption in the third step that goes from a defined daily doses per one million inhabitants of 897.67 in 2003 in area 5 and 1056.42 in area 2 to 1490.15 and 1601.48, respectively in 2006 and the second step goes from 1545.25 and 1629.94 in 2003 to 1904.70 and 1730.66 in both areas in 2006. In the hospitals analyzed, there is a similar progression, with a gradual increase in the use of fentanyl and a trend towards reduction in the consumption of morphine. CONCLUSIONS: Our data reflect the increase in consumption of opioids, despite declining morphine. It is noteworthy that transdermal fentanyl is the opioid prescribed most. PMID- 19121264 TI - [Risk factors, etiology and prognosis in patients with ischemic stroke and diabetes mellitus]. AB - INTRODUCTION: The objective of this work is to describe the clinical, etiological and prognostic differences in diabetic and non-diabetic patients presenting ischemic stroke. MATERIALS AND METHODS: We performed a retrospective review of all patients with ischemic stroke requiring admission to the Neurology Service at the University Hospital of Gran Canaria Dr. Negrin (Spain) during our study period (1 Jan, 2004 -31 Dec, 2006). We collected demographic, clinical and prognostic variables. RESULTS: A total of 633 patients were included, 256 (40.4%) were diabetic and 377 (59.6%) were non-diabetic. The group of diabetic patients presented a mean age of 69.2 < or = 9.3 years (65.4 < or = 13.9 years in non diabetic). Mean made up 63.7% of the diabetic patients and women 36.3% (62.2% men and 37.8% women in non-diabetics). A total of 85.9% of diabetic patients presented arterial hypertension (62.9% in non-diabetics); 49.2% had previous hyperlipidemia (31% in non-diabetics); 22% had ischemic heart disease (12.7% in nondiabetics); and 8.6% had peripheral arteriopathy (4.8% in non-diabetics). In the group of diabetic patients, 25% presented atherothrombotic stroke (16.4% in non-diabetics); 20.3% were cardioembolic (24.4% in non-diabetics); 26.1% were lacunar (22% in non-diabetics); and 25.7% had undetermined origin (27.2% in non diabetics). The modified Rankin Scale at discharge was +/- in 65.3% of diabetic patients and 62.5% in non-diabetics. CONCLUSIONS: In our series, diabetic patients frequently present other conventional vascular risk factors and show a tendency to more atherothrombotic strokes. We have found no significant differences in relation to prognosis. PMID- 19121265 TI - [Profitability of the bronchoscopy in the diagnosis of focal pulmonary malignant lesions]. AB - INTRODUCTION: We define focal pulmonary lesion (FPL) as an intra-parenchymatous pulmonary lesion that is well circumscribed and completely surrounded by healthy lung. It is considered that the profitability of the fine needle aspiration puncture (FNAP) in FPL < or = 2 cm is better than that of the fibrobronchoscopy (FBC). OBJECTIVE: To analyze the diagnostic profitability of the FNAP in the malignant FPL and study if it varies according to site, size and histology. MATERIAL AND METHODS: We analyzed all the FBCs of our Unit between 01/2000 and 12/2001 in patients with solitary FLP < or = 6 cm with a definitive diagnosis of malignancy. The diagnostic profitability by size, site and histology was analyzed with Pearson's chi(2) statistics. RESULTS: 124 patients. Mean FBC per patient was 1.3. A total of 101 cases (82%) were diagnosed with FBC, 15 by thoracotomy and 8 by FNAP. Global diagnostic profitability of the FBC was 0.82 and the transbronchial biopsy 0.76. There are no diagnostic profitability differences by size (< or = 2 cm vs > 2 cm) (0.81 vs 0.82 p = 0.96), site (peripheral vs central) (0.79 vs 0.85 p = 0.41) and histology (epidermoid vs adenocarcinoma) (0.89 vs 0.75 p = 0.21). CONCLUSION: Profitability of the FBC in malignant FPL in our hospital is elevated without differences by size, site or histology. In our site, the initial diagnostic approach of the FLP is done with FBC. PMID- 19121266 TI - [Serious adverse events derived from the drug interactions of antiretroviral therapy]. AB - BACKGROUND AND AIM: Antiretroviral drugs can cause drug interactions. MATERIAL AND METHODS: Three clinical cases are described regarding HIV-infected patients in which a clinically relevant adverse effect occurred due to a pharmacokinetic interaction. RESULTS: Case 1: A 43-year old woman being treated with tenofovir DF, emtricitabine and lopinavir/ritonavir who presents ischemia in both upper extremities following an ergotamine syndrome. Case 2: A 54-year old man being treated with zidovudine, lamivudine and lopinavir/ritonavir who presents Cushing syndrome following to use of inhaled fluticasone. Case 3: A 45-year old man being treated with tenofovir DF, emtricitabine and atazanavir/ritonavir who presents a virological failure as consequence of concomitant use of omeprazole. CONCLUSIONS: Potential drug interactions must be considered when other concomitant drugs are used with antiretroviral therapy especially when one of these is a P 450 cytochrome enzymatic inductor or inhibitor. PMID- 19121267 TI - [Rapid diagnosis unit in a third level hospital. Descriptive study of the first year and a half]. AB - BACKGROUND AND AIM: We show a descriptive study of a Rapid Diagnosis Unit as an alternative of the traditional hospital admission. MATERIAL AND METHODS: An Internist, a Radiologist and a Nurse have constituted Rapid Diagnosis Unit. A retrospective and descriptive study of 1132 patients visited in the first and a half- year is described. RESULTS: 1132 patients have been attended, 71% coming from Emergency Department and 26% from Primary Health Care. Mean stay was 9 days. Anaemia, wasting syndrome and lymph nodes enlargement were the most likely complaints. The outcome of these patients was diverse: 38% have been discharged, 52% carried on as outpatients in our hospital and 10% required hospitalization. In 199 patients (18%) cancer was diagnosed. Colon, lymphoma and lung cancer were the most common malignant disorders involved. About 4,5 beds per day have been saved from the Internal Medicine Department. CONCLUSIONS: Rapid Diagnosis Unit is an efficient alternative to conventional hospital admissions for patients in study of potential and serious disorders allowing an out-patient management. PMID- 19121268 TI - [Febrile syndrome in an elderly patient]. AB - Fever in the elderly as compared with young adults has a clinical expression, prevalence, and diagnosis that require a specific type of evaluation. The significance of fever in the elderly gains importance with the increase of the population's age and it must be considered an alarm signal in these patients. Etiologically, the primary cause of prolonged fever with unspecific origin in this population, is arteritis of giant cells, followed by (according to epidemiological environment) tuberculosis and hematological neoplasias. There is no consensus on diagnostic approaches, but it is essential to take great care in the process due to risk associated with invasive tests and low tolerance to them. PMID- 19121269 TI - [Non-invasive imaging techniques of the coronary tree]. AB - The technological development has made it possible to effectively evade the difficulties involved in the non-invasive study of the coronary tree, which is its small diameter and continuous movement. Improvement in the spatial and temporal resolution of the cardiac multidetector computed tomography has made it possible to incorporate this technique into the diagnostic armamentarium of coronary disease, with better results than the conventional tests. Its principal limitation is the need to use contrast and ionizing radiation. The cardiac magnetic resonance makes it possible to visualize the proximal coronary origins and segments without needing to use contrast or radiation. This makes it a reference technique in experienced sites when there is suspicion of a coronary abnormality, even as a <> technique in at risk populations, giving that it is innocuous. PMID- 19121270 TI - [Interstitial cells of Cajal: another contribution to modern medicine]. AB - The contributions of Santiago Ramon y Cajal to Modern medicine are innumerable. This large scientist and humanist, a man ahead of his time, spoke for the first time such concepts as angiogenesis, which represents one of the most innovative fields for oncology today, in addition to conducting experimental research on the degeneration and regeneration of the nervous system by which laid the foundation morphofunctional of neuroscience today. In this case, we wanted to remember the significance of the discovery of so-called <>, which have been involved in multiple processes and digestive diseases we collect, and especially his recent relationship with gastrointestinal stromal tumors with the therapeutic implications this could bring. PMID- 19121272 TI - [Ventricular pseudotachycardia]. PMID- 19121271 TI - [Fever and skin lesions in patients treated with phenobarbital]. PMID- 19121273 TI - [Uncommon cause of dysphagia in the elderly patient]. PMID- 19121274 TI - [A 64-year old woman with pulmonary nodule and arterial hypertension]. PMID- 19121275 TI - [Clinicopathological conferences of the SEMI]. PMID- 19121276 TI - [Predictive factors of severe disease among patients who consult to Emergency due to fever]. PMID- 19121277 TI - [Acute rhabdomyolysis and renal failure associated to status asthmaticus]. PMID- 19121278 TI - [Hypocomplementemia in the obstetric antiphospholipid syndrome]. PMID- 19121281 TI - Automated detection of haemozoin-containing monocytes for the diagnosis of malaria in microscopically negative cases during pregnancy. AB - Plasmodium falciparum sequesters in the placenta. Cell-Dyn automated flow cytometric haematology analysers have the capacity to detect haemozoin-containing circulating leukocytes during routine FBC analysis. In Lambarene, Gabon, 685 FBCs of pregnant women were analysed, yielding 86.8% sensitivity and 78.5% specificity compared to microscopy. In a subset of 37 Cell-Dyn positive but microscopy negative samples, PCR detected five positive cases. This methodology may serve as an adjunct rapid diagnostic tool for malaria during pregnancy, even in microscopically negative cases. PMID- 19121282 TI - H6PDH interacts directly with 11beta-HSD1: implications for determining the directionality of glucocorticoid catalysis. AB - Tissue specific amplification of glucocorticoid action through NADPH-dependent reduction of inactive glucocorticoid precursors by 11beta-hydroxysteroid dehydrogenase (11beta-HSD1) contributes to the development of visceral obesity, insulin resistance and Type 2 Diabetes. Hexose-6-phosphate dehydrogenase (H6PDH) is believed to supply NADPH for the reductase activity of 11beta-HSD1 in the lumen of the endoplasmic reticulum (ER), where the two enzymes are co-localized. We report here expression and purification of full-length and truncated N terminal domain (NTD) of H6PDH in a mammalian expression system. Interestingly, both full-length H6PDH and the truncated NTD are secreted into the culture medium in the absence of 11beta-HSD1. Purified full-length H6PDH is a bi-functional enzyme with glucose-6-phosphate dehydrogenase (G6PDH) activity as well as 6 phosphogluconolactonase (6PGL) activity. Using co-immunoprecipitation experiments with purified H6PDH and 11beta-HSD1, and with cell lysates expressing H6PDH and 11beta-HSD1, we observe direct physical interaction between the two enzymes. We also show the modulation of 11beta-HSD1 directionality by H6PDH using overexpression and siRNA knockdown systems. The NTD retains the ability to interact with 11beta-HSD1 physically as well as modulate 11beta-HSD1 directionality indicating that the NTD of H6PDH is sufficient for the regulation of the 11beta-HSD1 activity. PMID- 19121283 TI - Biochemical characterization and homology modeling of a purine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus: insights into mechanisms of protein stabilization. AB - We report the biochemical and structural characterization of the purine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus (SsIAG-NH). SsIAG-NH is a homodimer of 70kDa specific for adenosine, guanosine and inosine. SsIAG-NH is highly thermophilic and is characterized by extreme thermodynamic stability (T(m), 107 degrees C), kinetic stability and remarkable resistance to guanidinium chloride-induced unfolding. A disulfide bond that, on the basis of SDS-PAGE is positioned intersubunits, plays an important role in thermal stability. SsIAG-NH shares 43% sequence identity with the homologous pyrimidine specific nucleoside hydrolase from S. solfataricus (SsCU-NH). The comparative sequence alignment of SsIAG-NH, SsCU-NH, purine non-specific nucleoside hydrolase from Crithidia fasciculata and purine-specific nucleoside hydrolase from Trypanosoma vivax shows that, only few changes in the base pocket are responsible for different substrate specificity of two S. solfataricus enzymes. The structure of SsIAG-NH predicted by homology modeling allows us to infer the role of specific residues in substrate specificity and thermostability. PMID- 19121284 TI - Tight junctions and the regulation of gene expression. AB - Cell adhesion is a key regulator of cell differentiation. Cell interactions with neighboring cells and the extracellular matrix regulate gene expression, cell proliferation, polarity and apoptosis. Apical cell-cell junctions participate in these processes using different types of proteins, some of them exhibit nuclear and junctional localization and are called NACos for Nuclear Adhesion Complexes. Tight junctions are one type of such cell-cell junctions and several signaling complexes have been identified to associate with them. In general, expression of tight junction components suppresses proliferation to allow differentiation in a coordinated manner with adherens junctions and extracellular matrix adhesion. These tight junction components have been shown to affect several signaling and transcriptional pathways, and changes in the expression of tight junction proteins are associated with several disease conditions, such as cancer. Here, we will review how tight junction proteins participate in the regulation of gene expression and cell proliferation, as well as how they are regulated themselves by different mechanisms involved in gene expression and cell differentiation. PMID- 19121285 TI - Apolipoprotein E peptide-modified colloidal carriers: the design determines the mechanism of uptake in vascular endothelial cells. AB - Supramolecular structures, particularly micelles and liposomes equipped with uptake-mediating address compounds, have attracted much attention as pharmaceutical formulations. Their development requires an understanding of the mechanism by which the carrier systems interact with and translocate into the target cells. We developed an apolipoprotein E-derived peptide, called A2, that efficiently translocates across cell membranes. Upon coupling of two palmitoyl chains (P2), the highly cationic sequence acquires detergent-like properties such as a strong tendency to self-associate and the ability to integrate into lipid bilayers. Confocal laser scanning microscopy and fluorescence activated cell sorting were used to compare the internalization of the fluorescence-labeled monomeric A2 with the uptake of the colloidal P2A2 micelles and P2A2-tagged liposomes into endothelial cells of blood vessels. Specific inhibitors of endocytosis were used to identify the underlying mechanisms. b.End3 and BAEC cells as example of endothelial cells of small capillaries and large vascular vessels, respectively, were examined. The uptake of monomeric A2 was characterized by poor cellular selectivity. A2 was efficiently internalized into both cell lines via at least two different mechanisms. Besides an endocytotic uptake route, a second passive pathway exists, that leads to a rapid distribution of A2 within the cytoplasm. Also liposomes tagged with P2A2 were non-selectively internalized into both b.End3 and BAEC cells. Their nonselective uptake was mediated by clathrin- and caveolin-independent endocytosis. In contrast, micellar P2A2 entered b.End3 cells via clathrin-mediated endocytosis, while no uptake of P2A2 into BAEC cells was observed. In conclusion, the specific clathrin-mediated uptake mode of P2A2 micelles might provide the basis for a blood brain barrier specific targeting. PMID- 19121287 TI - MMSET is overexpressed in cancers: link with tumor aggressiveness. AB - MMSET is expressed ubiquitously in early development and its deletion is associated with the malformation syndrome called Wolf-Hirschhorn syndrome. It is involved in the t(4;14) (p16;q32) chromosomal translocation, which is the second most common translocation in multiple myeloma (MM) and is associated with the worst prognosis. MMSET expression has been shown to promote cellular adhesion, clonogenic growth and tumorigenicity in multiple myeloma. MMSET expression has been recently shown to increase with ascending tumor proliferation activity in glioblastoma multiforme. These data demonstrate that MMSET could be implicated in tumor emergence and/or progression. Therefore, we compared the expression of MMSET in 40 human tumor types--brain, epithelial, lymphoid--to that of their normal tissue counterparts using publicly available gene expression data, including the Oncomine Cancer Microarray database. We found significant overexpression of MMSET in 15 cancers compared to their normal counterparts. Furthermore MMSET is associated with tumor aggressiveness or prognosis in many types of these aforementioned cancers. Taken together, these data suggest that MMSET potentially acts as a pathogenic agent in many cancers. The identification of the targets of MMSET and their role in cell growth and survival will be key to understand how MMSET is associated with tumor development. PMID- 19121286 TI - Flotillin-1 stabilizes caveolin-1 in intestinal epithelial cells. AB - Flotillins and caveolins represent two types of resident proteins associated with lipid rafts in mammalian cells, however, their possible cross-talk in regulating lipid raft functions remains poorly understood. In this report, we observed that siRNA-mediated down-regulation of flotillin-1 expression which disrupted lipid raft-mediated endocytosis of BODIPY FL C(5)-lactosylceramide also substantially decreased caveolin-1 level in SK-CO15 human intestinal epithelial cells. The decrease in caveolin-1 expression appeared to be specific for flotillin-1 knock down and was not observed after down-regulation of flotillin-2. The decrease in caveolin-1 level in flotillin-1-depleted cells was not due to suppression of its mRNA synthesis and was not mimicked by cholesterol depletion of SK-CO15 cells. Furthermore, flotillin-1 dependent down-regulation of caveolin-1 was reversed after cell exposure to lysosomal inhibitor, chloroquine but not proteosomal inhibitor, MG262. Our data suggest that flotillin-1 regulates caveolin-1 level by preventing its lysosomal degradation in intestinal epithelial cells. PMID- 19121288 TI - Hepatic overexpression of dominant negative Mlx improves metabolic profile in diabetes-prone C57BL/6J mice. AB - Mlx and ChREBP form a heterodimer to regulate glucose-mediated gene expression in the liver. This study was performed to determine if the metabolic syndrome might be improved using dominant negative Mlx (dnMlx). An adenovirus bearing dnMlx was constructed and used to test the inhibitory effect of dnMlx on lipogenesis both in vitro and in vivo. Adenoviral overexpression of dnMlx in rat hepatocytes inhibited expression of glucose-regulated genes, including Chrebp and Transketolase, which constitute a positive feedback loop in the regulation of Chrebp gene expression. Adenoviral overexpression of dnMlx in 25-week-old male C57BL/6J mice reduced hepatic triglyceride contents and improved glucose intolerance by inhibiting expression of Glucose-6-phosphatase and Elovl6 mRNA in addition to lipogenic enzymes. In conclusion, overexpression of dnMlx improves glucose intolerance by inhibiting expression not only of lipogenic enzymes but also other important genes such as Glucose-6-phosphatase and Elovl6. PMID- 19121289 TI - Half-of-the-sites reactivity of transketolase from Saccharomyces cerevisiae. AB - Cleavage by yeast transketolase of the donor substrate, D-xylulose 5-phosphate, in the absence of the acceptor substrate was studied using stopped-flow spectrophotometry. One mole of the substrate was shown to be cleaved in the prestationary phase, leading to the formation of one mole of the reaction product per mole enzyme, which has two active centers. This observation indicates that only one out of the two active centers functions (i.e., binds and cleaves the substrate) at a time. Such half-of-the-sites reactivity of transketolase conforms well with our understanding, proposed previously, that the active centers of the enzyme operate in sequence (in phase opposition): the cleavage of a ketose within one center (first phase of the transketolase reaction) is paralleled by its formation in the other center (glycolaldehyde residue is condensed with the acceptor substrate, and the second stage of the transketolase reaction is thereby completed) [M.V. Kovina, G.A. Kochetov, FEBS Lett. 440 (1998) 81-84]. PMID- 19121290 TI - An EGF-like peptide sequence from Dictyostelium enhances cell motility and chemotaxis. AB - Dictyostelium discoideum possesses more EGF-like (EGFL) domains than any other sequenced eukaryote. Here we show that a synthetic EGFL peptide (DdEGFL1) based upon an amino acid sequence from a cysteine-rich Dictyostelium protein, functions extracellularly to enhance random cell motility and cAMP-mediated chemotaxis in Dictyostelium by 625% and 85%, respectively, in strain NC4 and by 620% and 80% in strain AX3. Quinacrine inhibited peptide-enhanced random motility but not chemotaxis in strain AX3 providing evidence that PLA2 is the predominant regulator of this process. While LY294002 alone had no significant effect on either event, in combination with quinacrine it dramatically inhibited both processes suggesting that both PI3K and PLA2-mediated signaling are required for EGFL peptide-enhanced cell movement. DdEGFL1 also sustained the threonine phosphorylation of a 210kDa protein that is dephosphorylated during Dictyostelium starvation. Taken together, these results suggest an important role for certain EGFL peptides in Dictyostelium cell movement. PMID- 19121291 TI - Efficient lowering of triglyceride levels in mice by human apoAV protein variants associated with hypertriglyceridemia. AB - Variation in the apolipoprotein A5 (APOA5) gene has consistently been associated with increased plasma triglyceride (TG) levels in epidemiological studies. In vivo functionality of these variations, however, has thus far not been tested. Using adenoviral over-expression, we evaluated plasma expression levels and TG lowering efficacies of wild-type human apoAV, two human apoAV variants associated with increased TG (S19W, G185C) and one variant (Q341H) that is predicted to have altered protein function. Injection of mice with adenovirus encoding wild-type or mutant apoAV resulted in an identical dose-dependent elevation of human apoAV levels in plasma. The increase in apoAV levels resulted in pronounced lowering of plasma TG levels at two viral dosages. Unexpectedly, the TG-lowering efficacy of all three apoAV variants was similar to wild-type apoAV. In addition, no effect on TG-hydrolysis-related plasma parameters (free fatty acids, glycerol and post heparin lipoprotein lipase activity) was apparent upon expression of all apoAV variants. In conclusion, our data indicate that despite their association with hypertriglyceridemia and/or predicted protein dysfunction, the 19W, 185C and 341H apoAV variants are equally effective in reducing plasma TG levels in mice. PMID- 19121292 TI - Inhibitory study of protein arginine methyltransferase 1 using a fluorescent approach. AB - Protein arginine methyltransferases (PRMTs) play important roles in both normal physiology and human diseases. Deregulation of PRMT activity has been linked to several pathological states such as cancer and cardiovascular disorders. Herein, we report our work of designing and using new fluorescent reporters to perform single-step analysis of substrate binding and methylation by PRMT1. Both fluorescence intensity and anisotropy of the two reporters, R4-FL and H4-FL, were shown to effectively manifest enzyme-substrate interactions, highlighting their application in investigating PRMT inhibitors. In particular, the methylation process of R4-FL can be directly studied using fluorescence intensity readout. By combining the fluorescent measurement with radioactive analysis, we determined that AMI-1 inhibits PRMT1 activity through the mechanism of blocking peptide substrate binding. PMID- 19121293 TI - Cholestane-3beta,5alpha,6beta-triol-induced reactive oxygen species production promotes mitochondrial dysfunction in isolated mice liver mitochondria. AB - Reactive oxygen species (ROS) have been implicated in oxysterol-induced apoptosis. However, the mechanism of ROS production induced by oxysterols within cells is not clear. Considering that mitochondria is the main source of intracellular ROS, and play a key role in oxysterol-induced apoptosis, we investigated the effect of oxysterol cholestane-3beta,5alpha,6beta-triol (Triol) on ROS production and mitochondrial function in isolated mice liver mitochondria. Triol at higher concentrations (10-50 microM) enhanced the production of O(2)(-) and H(2)O(2) in isolated mitochondria, which might be due to its stimulation to the activities of complexes I and II of mitochondrial electron transfer chain, and its inhibition to glutathione peroxidase activity. The same concentrations of Triol induced obviously oxidative damage of mitochondrial membrane lipids and proteins, as demonstrated by the increased MDA level and the decreased protein thiols content. Furthermore, Triol caused mitochondrial dysfunction, including the opening of mitochondrial permeability transition pore, the decrease of mitochondrial membrane potential (DeltaPsi(m)), and the release of cytochrome c. Antioxidant butylated hydroxytoluene significantly inhibited oxidative damage, the decrease of DeltaPsi(m), and the release of cytochrome c, implying that ROS might mediate mitochondrial dysfunction induced by Triol. We concluded that Triol induced mitochondrial ROS production and subsequently oxidative damage, leading to the mitochondrial dysfunction, thus suggesting a putative mechanism of apoptosis activation by oxysterols in vascular cells. PMID- 19121294 TI - 5,6-Dihydrocyclopenta[c][1,2]-dithiole-3(4H)-thione is a promising cancer chemopreventive agent in the urinary bladder. AB - It has been widely recognized that induction of Phase 2 enzymes is an effective and sufficient strategy for achieving protection against carcinogenesis. Nrf2 is the unifying master regulator of these enzymes and its activation in various tissues, including the urinary bladder, is associated with inhibition of carcinogenesis. 5,6-Dihydrocyclopenta[c][1,2]-dithiole-3(4H)-thione (CPDT) is a highly potent inducer of Phase 2 enzymes and an activator of Nrf2. In vivo, it is particularly effective in the bladder, showing significant effects in this tissue when dosed to rats at levels as low as 0.98 micromol/(kgday) (0.17 mg/(kg day)). The activities of key Phase 2 enzymes, including glutathione S-transferase, NAD(P)H:quinone:oxidoreductase 1 and glutamate cysteine synthetase, and levels of glutathione were elevated by CPDT in rat bladder in vivo and in cultured bladder cells in vitro. In the bladder, enzyme induction and Nrf2 activation appear to occur exclusively in the epithelium. This is highly significant, since almost all bladder cancers develop from the epithelium. Studies in cultured bladder cells using siRNA to knock down Nrf2 or in cells with total Nrf2 knockout showed that the ability of CPDT to induce Phase 2 enzymes depends completely on Nrf2. In conclusion, CPDT potently and preferentially induces Phase 2 enzymes in the bladder epithelium and Nrf2 is its key mediator. PMID- 19121295 TI - Anticancer activity of an essential oil from Cymbopogon flexuosus. AB - The essential oil from a lemon grass variety of Cymbopogon flexuosus was studied for its in vitro cytotoxicity against twelve human cancer cell lines. The in vivo anticancer activity of the oil was also studied using both solid and ascitic Ehrlich and Sarcoma-180 tumor models in mice. In addition, the morphological changes in tumor cells were studied to ascertain the mechanism of cell death. The in vitro cytotoxicity studies showed dose-dependent effects against various human cancer cell lines. The IC(50) values of oil ranged from 4.2 to 79 microg/ml depending upon the cell line. In 502713 (colon) and IMR-32 (neuroblastoma) cell lines, the oil showed highest cytotoxicity with IC(50) value of 4.2 and 4.7 microg/ml, respectively. Intra-peritoneal administration of the oil significantly inhibited both ascitic and solid forms of Ehrlich and Sarcoma-180 tumors in a dose-dependent manner. The tumor growth inhibition at 200 mg/kg (i.p.) of the oil observed with both ascitic and solid tumor forms of Ehrlich Ascites carcinoma was 97.34 and 57.83 respectively. In case of Sarcoma-180, the growth inhibition at similar dose of oil was 94.07 and 36.97% in ascitic and solid forms respectively. Morphological studies of the oil treated HL-60 cells revealed loss of surface projections, chromatin condensation and apoptosis. The mitochondria showed apparent loss of cristae in the cells undergoing apoptosis. The morphological studies of Sarcoma-180 solid tumor cells from animals treated with the oil revealed condensation and fragmentation of nuclei typical of apoptosis. Morphological studies of ascites cells from animals treated with the oil too revealed the changes typical of apoptosis. Our results indicate that the oil has a promising anticancer activity and causes loss in tumor cell viability by activating the apoptotic process as identified by electron microscopy. PMID- 19121296 TI - Effects of Artemisia capillaris ethyl acetate fraction on oxidative stress and antioxidant enzyme in high-fat diet induced obese mice. AB - The current study examined the effects of Artemisia capillaris ethyl acetate (ACE) fraction on oxidative stress and antioxidant enzyme in high-fat diet induced obese mice. C57BL/6J mice were randomly assigned to normal diet group and high-fat diet groups. Normal diet groups were fed with an ACE fraction-free diet or 0.1 g ACE fraction/kg body weight. The high-fat diet groups were fed with an ACE fraction-free diet, 0.05 g ACE fraction, 0.1 g ACE fraction or 0.03 g rosiglitazone/kg body weight. ACE fraction strengthened the antioxidative defense system with an increased activity of superoxide dismutase (SOD) and decreased activities of glutathione-peroxidase (GSHpx) and catalase (CAT) and content of reduced glutathione (GSH). Accordingly, ACE fraction was found to reduce the accumulation of reactive oxygen species (ROS), such as superoxide radical and hydrogen peroxide which decrease the generation of oxidative stress damage substrates, such as the thiobarbituric acid reactive substances (TBARS) and carbonyl value. ACE fraction was found to be excellent for strengthening the antioxidant defense system, reducing the generation of ROS and damaging oxidative substances in the liver of high-fat diet induced obese mice. PMID- 19121297 TI - Amino acids levels and lipid peroxidation in maple syrup urine disease patients. AB - OBJECTIVE: In the present study we correlated the amino acids, branched-chain alpha-keto acids and alpha-hydroxy acids levels with the thiobarbituric acid reactive species (TBARS) measurement, a lipid peroxidation parameter, in plasma from treated MSUD patients in order to examine whether these accumulated metabolites could be associated to the oxidative stress present in MSUD. DESIGN AND METHODS: TBARS, amino acids, branched-chain alpha-keto acids and alpha hydroxy acids concentrations were measured in plasma samples from treated MSUD patients. RESULTS: We verified that plasma TBARS was increased, whereas tryptophan and methionine concentrations were significantly reduced. Furthermore TBARS measurement was inversely correlated to methionine and tryptophan levels. CONCLUSIONS: Considering that methionine and tryptophan have antioxidant activities, the data suggest that the imbalance of these amino acids may be involved with lipid peroxidation in MSUD. PMID- 19121298 TI - MUC1 568 A/G genotype-dependent cancer antigen 15-3 levels in breast cancer patients. AB - OBJECTIVES: CA 15-3 is a widely used tumor marker for breast cancer. We have investigated whether the MUC1 568 A/G polymorphism can influence CA 15-3 levels in healthy women and patients with breast tumors. DESIGN AND METHODS: CA 15-3 was measured in 208 healthy women, in 67 with benign disease, and in 162 women with breast cancer. All subjects were genotyped for the MUC1 568 A/G polymorphism. RESULTS: Significant differences were observed between mean CA 15-3 levels of control subjects grouped according to the MUC1 568 genotype (mean+/-SD): AA (10.3+/-3.8), AG (15.9+/-5.0) and GG (19.0+/-5.6) U/mL, p<0.0001. Similar (median) results were observed in women with benign breast disease: AA (10.2), AG (14.2) and GG (16.6) U/mL, p<0.0001, and those with breast cancer: AA (10.4), AG (17.1) and GG (23.9) U/mL, p<0.0001. CONCLUSIONS: The MUC1 568 A/G polymorphism strongly influences CA 15-3 levels in healthy women and women with either benign or malignant breast tumors. PMID- 19121299 TI - Ice-binding proteins from enoki and shiitake mushrooms. AB - Fungi have developed a variety of mechanisms for tolerating cold, including production of proteins that bind to ice, as shown by their ability to slightly lower the freezing point. At present, only one of these proteins, from the snow mold Typhula ishikariensis, and partial transcripts of a similar protein from shiitake mushroom, Lentinula edodes, have been identified. Here, we report the full sequences of ice-binding proteins from shiitake and another mushroom, the cold-adapted Flammulina populicola (enoki mushroom), and show that the recombinant proteins have ice-binding activity. The three proteins share 50-55% identities and are similar to other ice-binding proteins recently identified in ice bacteria and sea ice diatoms. The possibility that ice-binding protein genes have spread among these phyla by horizontal transfer is discussed. PMID- 19121301 TI - The specific targeting of guidance receptors within neurons: who directs the directors? AB - Guidance molecules present in both axonal and dendritic growth cones mediate neuronal responses to extracellular cues thereby ensuring correct neurite pathfinding and development of the nervous system. Little is known though about the mechanisms employed by neurons to deliver these receptors, specifically and efficiently, to the extending growth cone. A deeper understanding of this process is crucial if guidance receptors are to be manipulated to promote nervous system repair. Studies in other polarised cells, notably epithelial, have elucidated fundamental routes to the intracellular segregation of molecules mediated by endosomal pathways. Due to their extreme complexity and specialisation, neurons appear to have built upon these generic systems to evolve sophisticated trafficking networks. A striking feature is the axon initial segment which acts like a valve to tightly regulate the flux of molecules both entering and leaving the axon. Once in the growth cone, further controls operate to enhance the retention or rejection, as appropriate, of membrane receptors. We discuss the current state of knowledge regarding the intracellular trafficking of axon guidance receptors and how this relates to their developmental roles. We highlight the various facets still to be properly elucidated and by building on existing data regarding neuronal polarity and intracellular sorting mechanisms suggest ways to fill these gaps. PMID- 19121300 TI - Morphogenetic movements driving neural tube closure in Xenopus require myosin IIB. AB - Vertebrate neural tube formation involves two distinct morphogenetic events- convergent extension (CE) driven by mediolateral cell intercalation, and bending of the neural plate driven largely by cellular apical constriction. However, the cellular and molecular biomechanics of these processes are not understood. Here, using tissue-targeting techniques, we show that the myosin IIB motor protein complex is essential for both these processes, as well as for conferring resistance to deformation to the neural plate tissue. We show that myosin IIB is required for actin-cytoskeletal organization in both superficial and deep layers of the Xenopus neural plate. In the superficial layer, myosin IIB is needed for apical actin accumulation, which underlies constriction of the neuroepithelial cells, and that ultimately drive neural plate bending, whereas in the deep neural cells myosin IIB organizes a cortical actin cytoskeleton, which we describe for the first time, and that is necessary for both normal neural cell cortical tension and shape and for autonomous CE of the neural tissue. We also show that myosin IIB is required for resistance to deformation ("stiffness") in the neural plate, indicating that the cytoskeleton-organizing roles of this protein translate in regulation of the biomechanical properties of the neural plate at the tissue-level. PMID- 19121302 TI - Gli3 is required autonomously for dorsal telencephalic cells to adopt appropriate fates during embryonic forebrain development. AB - The Gli3 zinc finger transcription factor is expressed in developing forebrain, with the highest levels of expression in dorsal telencephalon. In Gli3(-/-) embryos the dorsal telencephalon is abnormally small and fails to develop dorsomedial telencephalic structures, including hippocampus and cortical hem, while the ventral telencephalon appears to expand. A hurdle to understanding the underlying mechanisms is that abnormalities of developing Gli3(-/-) telencephalic cells in Gli3(-/-) mutants result from a combination of their own cell autonomous defects and defects in the Gli3(-/-) cells that surround them. Here we used chimeras to identify some of the defects of Gli3(-/-) telencephalic cells that are likely to be autonomous by studying how Gli3(-/-) cells develop when surrounded by a majority of wild-type cells. We found that Gli3(-/-) cells are present in all components of the Gli3(-/-)<-->Gli3(+/+) chimeric forebrain, including dorsomedial structures, in proportions that either equal or exceed proportions found elsewhere in the embryo. Gli3(-/-) cells segregate from Gli3(+/+) cells to form many abnormal structures particularly in dorsal telencephalon. Gli3(-/-) cells in some locations are misspecified: in those parts of the dorsal telencephalon near to its boundaries with the diencephalon and the ventral telencephalon, mutant cells express sets of transcription factors expressed by wild-type cells on the other side of the boundary. Elsewhere in the dorsal telencephalon, in the diencephalon and in the ventral telencephalon, mutant cells express sets of transcription factors similar to those expressed by their immediately surrounding wild-type cells. We propose that an important cell autonomous action of Gli3 is to regulate the competence of dorsal telencephalic cells, preventing cells near to its boundaries expressing regulatory factors normally restricted to adjacent tissues. PMID- 19121303 TI - Three distinct RNA localization mechanisms contribute to oocyte polarity establishment in the cnidarian Clytia hemisphaerica. AB - Egg animal-vegetal polarity in cnidarians is less pronounced than in most bilaterian species, and its normal alignment with the future embryonic axis can be disturbed by low-speed centrifugation. We have analyzed the development of oocyte polarity within the transparent and autonomously functioning gonads of Clytia medusae, focusing on the localization of three recently identified maternal mRNAs coding for axis-directing Wnt pathway regulators. Animal-vegetal polarity was first detectable in oocytes committed to their final growth phase, as the oocyte nucleus (GV) became positioned at the future animal pole. In situ hybridization analyses showed that during this first, microtubule-dependent polarization event, CheFz1 RNA adopts a graded cytoplasmic distribution, most concentrated around the GV. CheFz3 and CheWnt3 RNAs adopt their polarized cortical localizations later, during meiotic maturation. Vegetal localization of CheFz3 RNA was found to require both microtubules and an intact gonad structure, while animal localization of CheWnt3 RNA was microtubule independent and oocyte autonomous. The cortical distribution of both these RNAs was sensitive to microfilament-disrupting drugs. Thus, three temporally and mechanistically distinct RNA localization pathways contribute to oocyte polarity in Clytia. Unlike the two cortical RNAs, CheFz1 RNA was displaced in fertilized eggs upon centrifugation, potentially explaining how this treatment re-specifies the embryonic axis. PMID- 19121304 TI - The role of Notch signalling and numb function in mechanosensory organ formation in the spider Cupiennius salei. AB - In the spider Cupiennius salei the mechanosensory organs of the legs are generated from epithelial sensory precursor groups which are specified by elevated levels of the achaete-scute homologues CsASH1 and CsASH2. Neural precursors delaminate from the groups and occupy positions basal and proximal to the accessory cells which remain in the epithelium. Here we analyse the role of Notch signalling and numb function in the development of the mechanosensory organs of the spider. We show that Notch signalling is required for several processes: the selection of the sensory precursor groups, the maintenance of undifferentiated sensory precursors, the binary cell fate decision between accessory and neural fate and the differentiation of sensory neurons. Our data suggest that Numb antagonises Notch signalling in the neural precursors, which results in activation of the neural cell fate determinant Prospero and delamination of the neural precursors from the epithelium. Prospero is expressed de novo in sensory neural precursors and we assume that the expression of the gene is regulated by the Notch to Numb ratio within the sensory precursors. Interestingly, the spider numb RNAi phenotype resembles the numb/numblike loss of function phenotypes in the mammalian nervous system, indicating that the interaction between Notch signalling and Numb might play a similar role in both systems. PMID- 19121305 TI - Critical time-window for the actions of adrenal glucocorticoids in behavioural sensitisation to cocaine. AB - Glucocorticoids, secreted by the adrenals in response to stress, have profound effects on behavioural responsiveness to psychostimulant drugs. We have studied the critical time-window for the influence of corticosterone on behavioural sensitisation to cocaine in relation to i) the stage of behavioural sensitisation, and ii) the time of drug exposure. Previously, we have identified a mouse strain (DBA/2) in which surgical removal of the adrenals (adrenalectomy) fully prevented locomotor sensitisation to cocaine. To investigate the role of corticosterone in expression of behavioural sensitisation, the glucocorticoid receptor antagonist mifepristone (RU38486) was administered to previously sensitised mice prior to a cocaine challenge. Furthermore, adrenalectomised mice were given corticosterone replacement at different intervals prior to each drug administration, to investigate the role of the glucocorticoid in initiation of behavioural sensitisation, and in relation to the time of drug exposure. Administration of mifepristone to previously sensitised animals failed to block expression of cocaine-induced behavioural sensitisation. In adrenalectomised mice, intermittent replacement of corticosterone (1 mg/kg i.p., either 2 h or 5 min prior to each cocaine administration), did not reverse the sensitisation deficit. By contrast, chronic corticosterone replacement (20% pellet) partially restored initiation of behavioural sensitisation. These data indicate that the presence of corticosterone facilitates the initiation rather than the expression of behavioural sensitisation to cocaine. However, because high corticosterone concentrations only partially reversed the sensitisation deficit of adrenalectomised mice, the adrenal glucocorticoid seems necessary, but not sufficient, for full behavioural sensitisation to cocaine in the DBA/2 strain. PMID- 19121306 TI - WAFL, a new protein involved in regulation of early endocytic transport at the intersection of actin and microtubule dynamics. AB - We have previously identified a new gene with sequence homology to the WASP family of actin regulators denoted WAFL (WASP and FKBP-like). Here we report a possible biological function for WAFL, by demonstrating an association to early endosomes via its central coiled-coil domain. Further we show by functional and structural studies that WAFL is associated with both microtubules and the actin filament system, the two means of transport of early endosomes. In addition, WAFL interacts with WASP-interacting protein (WIP) and actin, thus linking WAFL to actin dynamics. The use of RNAi depletion of WAFL shows that WAFL-deficient cells display delayed transport of endosomal cargo. Our findings are compatible with a model whereby WAFL is involved in the transport of early endosomes at the level of transition between microfilament-based and microtubule-based movement. PMID- 19121307 TI - Type I interferon receptor signalling is induced during demyelination while its function for myelin damage and repair is redundant. AB - The type I interferons, interferon-beta and alpha (IFN-beta, IFN-alpha), are widely used for the treatment of autoimmune demyelination in the central nervous system (CNS). Their effects on de- and remyelination through the broadly expressed type I IFN receptor (IFNAR), however, are highly speculative. In order to elucidate the role of endogenous type I interferons for myelin damage and recovery we induced toxic demyelination in the absence of IFNAR1. We demonstrate that IFNAR signalling was induced during acute demyelination since the cytokine IFN-beta as well as the IFN-dependent genes IRF7, ISG15 and UBP43 were strongly upregulated. Myelin damage, astrocytic and microglia response, however, were not significantly reduced in the absence of IFNAR1. Furthermore, motor skills of IFNAR1-deficient animals during non-immune demyelination were unaltered. Finally, myelin recovery was found to be independent from endogenous IFNAR signalling, indicating a redundant role of this receptor for non-inflammatory myelin damage and repair. PMID- 19121308 TI - AtAzg1 and AtAzg2 comprise a novel family of purine transporters in Arabidopsis. AB - In plants, nucleobase biochemistry is highly compartmented relying upon a well regulated and selective membrane transport system. In Arabidopsis two proteins, AtAzg1 and AtAzg2, show substantial amino acid sequence similarity to the adenine guanine-hypoxanthine transporter AzgA of Aspergillus nidulans. Analysis of single and double mutant lines harboring T-DNA insertion alleles AtAzg1-1 and AtAzg2-1 reveal a marked resistance to growth in the presence of 8-azaadenine and 8 azaguanine but not to other toxic nucleobase analogues. Conversely, yeast strains expressing AtAzg1 and AtAzg2 gain heightened sensitivity to growth on 8 azaadenine and 8-azaguanine. Radio-labeled purine uptake experiments in yeast and in planta confirm the function of AtAzg1 and AtAzg2 as plant adenine-guanine transporters. PMID- 19121309 TI - Cold stability of intrinsically disordered proteins. AB - Contrary to globular proteins, intrinsically disordered proteins (IDPs) lack a folded structure and they do not lose solubility at elevated temperatures. Although this should also be true at low temperatures, cold stability of IDPs has not been addressed in any scientific work so far. As direct characterization of cold-denaturation is difficult, we approached the problem through a freezing induced loss-of-function model of globular-disordered functional protein pairs (m calpain-calpastatin, tubulin-Map2c, Hsp90-ERD14). Our results affirm that in contrast with globular proteins IDPs are resistant to cold treatment. The theoretical and functional aspects of this observation are discussed. PMID- 19121310 TI - Gibberellin biosynthesis in bacteria: separate ent-copalyl diphosphate and ent kaurene synthases in Bradyrhizobium japonicum. AB - Gibberellins are ent-kaurene-derived diterpenoid phytohormones produced by plants, fungi, and bacteria. The distinct gibberellin biosynthetic pathways in plants and fungi are known, but not that in bacteria. Plants typically use two diterpene synthases to form ent-kaurene, while fungi use only a single bifunctional diterpene synthase. We demonstrate here that Bradyrhizobium japonicum encodes separate ent-copalyl diphosphate and ent-kaurene synthases. These are found in an operon whose enzymatic composition indicates that gibberellin biosynthesis in bacteria represents a third independently assembled pathway relative to plants and fungi. Nevertheless, sequence comparisons also suggest potential homology between diterpene synthases from bacteria, plants, and fungi. PMID- 19121311 TI - The NtAMI1 gene functions in cell division of tobacco BY-2 cells in the presence of indole-3-acetamide. AB - Tobacco (Nicotiana tabacum) Bright Yellow-2 (BY-2) cells can be grown in medium containing indole-3-acetamide (IAM). Based on this finding, the NtAMI1 gene, whose product is functionally equivalent to the AtAMI1 gene of Arabidopsis thaliana and the aux2 gene of Agrobacterium rhizogenes, was isolated from BY-2 cells. Overexpression of the NtAMI1 gene allowed BY-2 cells to proliferate at lower concentrations of IAM, whereas suppression of the NtAMI1 gene by RNA interference (RNAi) caused severe growth inhibition in the medium containing IAM. These results suggest that IAM is incorporated into plant cells and converted to the auxin, indole-3-acetic acid, by NtAMI1. PMID- 19121312 TI - ApoC-III gene expression is sharply increased during adipogenesis and is augmented by retinoid X receptor (RXR) agonists. AB - ApoC-III, a lipoprotein lipase inhibitor and a key regulator of the metabolism of triglyceride-rich lipoproteins, is mainly synthesized in liver and intestine, but little is known about its expression in adipocytes. We here show that apoC-III mRNA levels are dramatically induced during adipocyte differentiation. Among many transcriptional factors involved in adipocyte differentiation, RXRalpha, acting alone and not as a heterodimer partner of other nuclear receptors, solely regulates apoC-III gene expression. These results suggest that apoC-III may play a specific role in lipid storage and mobilization in adipocytes, non-lipoprotein secreting cells, and indicate the functional role of RXRalpha during adipocyte differentiation. PMID- 19121313 TI - A possible role of vimentin on the cell surface for the activation of latent transforming growth factor-beta. AB - Latent TGF-beta (LTGF-beta) has to be converted to active TGF-beta for its activities. Previously, we reported that certain fragments of latency associated peptide (LAP) augmented LTGF-beta activation via increase in binding of LTGF-beta to the endothelial cell (EC) surface followed by cell-associated proteolysis. By searching for EC membrane proteins crosslinked with the LAP fragment, we identified the molecule bound to LAP fragment as vimentin. Moreover, the LAP fragment-induced LTGF-beta activation was attenuated by anti-vimentin antibody. These results indicate that binding of the LAP fragment to vimentin on the cell surface is indispensable for LTGF-beta activation by the LAP fragment. PMID- 19121314 TI - Acetyl-l-carnitine inhibits TNF-alpha-induced insulin resistance via AMPK pathway in rat skeletal muscle cells. AB - In this study, we demonstrated effects of acetyl-l-carnitine (ALC) on insulin resistance induced by tumor necrosis factor-alpha (TNF-alpha) in rat L6 cells. TNF-alpha downregulated insulin-stimulated glucose uptake and increased Serine 307 phosphorylation of insulin receptor substrate-1 (IRS-1). However, the treatment of ALC improved insulin-stimulated glucose uptake via AMP-activated protein kinase (AMPK) activation in a dose-dependent manner. Together, our data suggest that ALC inhibits TNF-alpha-induced insulin resistance through AMPK pathway in skeletal muscle cells. PMID- 19121315 TI - Clinical challenges and images in GI. Image 1: huge peritoneal loose body in the pelvic cavity. PMID- 19121317 TI - The interleukin-6 family cytokine interleukin-11 regulates homeostatic epithelial cell turnover and promotes gastric tumor development. AB - BACKGROUND & AIMS: Gastric cancer is the second most common cause of cancer related mortality worldwide, mainly as a result of late-stage detection. Interleukin (IL)-11 is a multifunctional cytokine reported to be up-regulated in human gastric cancer. METHODS: We investigated the importance of IL-11 in gastric cancer progression by examining its role in a variety of mouse gastric tumor models, as well as in nonneoplastic and tumor tissues taken from gastric cancer patients. We then determined the transcriptional and translational outcomes of IL 11 overexpression in normal gastric mucosa and identified a novel gene signature important early in the progression toward gastric tumorigenesis. RESULTS: IL-11 was up-regulated significantly in 4 diverse mouse models of gastric pathology as well as in human biopsy specimens adjacent to and within gastric cancer. Removal of IL-11 co-receptor alpha significantly reduced HKbeta-/- mouse fundic hyperplasia and ablated gp130(757F/F) mouse tumorigenesis. Exogenous IL-11 but not IL-6 activated oncogenic signal transducer and activator of transcription-3, and altered expression of novel proliferative and cytoprotective genes RegIII beta, RegIII-gamma, gremlin-1, clusterin, and growth arrest specific-1 in wild type gastric mucosa, a gene signature common in gp130(757F/F) and HKbeta-/- tumors as well as nonneoplastic mucosa of gastric cancer patients. One week of chronic IL-11 administration in wild-type mice sustained the gene signature, causing pretumorigenic changes in both antrum and fundus. CONCLUSIONS: Increased gastric IL-11 alters expression of proliferative and cytoprotective genes and promotes pretumorigenic cellular changes. PMID- 19121316 TI - Glutamine increases autophagy under Basal and stressed conditions in intestinal epithelial cells. AB - BACKGROUND & AIMS: Glutamine plays a protective role in intestinal cells during physiologic stress; however, the protection mechanisms are not fully understood. Autophagy functions in bulk degradation of cellular components, but has been recognized recently as an important mechanism for cell survival under conditions of stress. We therefore sought to see if glutamine's actions involve the induction of autophagy in intestinal cells and, if so, the mechanisms that underlie this action. METHODS: Formation of microtubule-associated protein light chain 3 (LC3)-phospholipid conjugates (LC3-II) in rat intestinal epithelial IEC 18 cells and human colonic epithelial Caco-2(BBE) cells was determined by Western blotting and localized by confocal microscopy. Activation of mammalian target of rapamycin (mTOR) pathway, mitogen-activated protein (MAP) kinases, caspase-3, and poly (ADP-ribose) polymerase were monitored by Western blotting. RESULTS: Glutamine increased LC3-II as well as the number of autophagosomes. Glutamine induced LC3-II formation was paralleled by inactivation of mTOR and p38 MAP kinase pathways, and inhibition of mTOR and p38 MAP kinase allowed LC3-II induction in glutamine-deprived cells. Under glutamine starvation, LC3-II recovery after heat stress or the increase under oxidative stress was blunted significantly. Glutamine depletion increased caspase-3 and poly (ADP-ribose) polymerase activity after heat stress, which was inhibited by treatment with inhibitors of mTOR and p38 MAP kinase. CONCLUSIONS: Glutamine induces autophagy under basal and stressed conditions, and prevents apoptosis under heat stress through its regulation of the mTOR and p38 MAP kinase pathways. We propose that glutamine contributes to cell survival during physiologic stress by induction of autophagy. PMID- 19121318 TI - Compound heterozygous mutations affect protein folding and function in patients with congenital sucrase-isomaltase deficiency. AB - BACKGROUND & AIMS: Congenital sucrase-isomaltase (SI) deficiency is an autosomal recessive intestinal disorder characterized by a drastic reduction or absence of sucrase and isomaltase activities. Previous studies have indicated that single mutations underlie individual phenotypes of the disease. We investigated whether compound heterozygous mutations, observed in some patients, have a role in disease pathogenesis. METHODS: We introduced mutations into the SI complementary DNA that resulted in the amino acid substitutions V577G and G1073D (heterozygous mutations found in one group of patients) or C1229Y and F1745C (heterozygous mutations found in another group). The mutant genes were expressed transiently, alone or in combination, in COS cells and the effects were assessed at the protein, structural, and subcellular levels. RESULTS: The mutants SI-V577G, SI G1073D, and SI-F1745C were misfolded and could not exit the endoplasmic reticulum, whereas SI-C1229Y was transported only to the Golgi apparatus. Co expression of mutants found on each SI allele in patients did not alter the protein's biosynthetic features or improve its enzymatic activity. Importantly, the mutations C1229Y and F1745C, which lie in the sucrase domains of SI, prevented its targeting to the cell's apical membrane but did not affect protein folding or isomaltase activity. CONCLUSIONS: Compound heterozygosity is a novel pathogenic mechanism of congenital SI deficiency. The effects of mutations in the sucrase domain of SIC1229Y and SIF1745C indicate the importance of a direct interaction between isomaltase and sucrose and the role of sucrose as an intermolecular chaperone in the intracellular transport of SI. PMID- 19121319 TI - Exposed: the genetic underpinnings of ulcerative colitis relative to Crohn's disease. PMID- 19121320 TI - Molecular characterization of ghrelin and gastrin-releasing peptide in Atlantic cod (Gadus morhua): cloning, localization, developmental profile and role in food intake regulation. AB - Full-length complementary deoxyribonucleic acid as well as genomic sequences encoding for two gastrointestinal appetite-related peptides, ghrelin and for gastrin-releasing peptide (GRP) were cloned from Atlantic cod (Gadus morhua) stomach using reverse transcription and rapid amplification of complementary deoxyribonucleic acid ends. Semi-quantitative reverse transcriptase polymerase chain reaction shows that both ghrelin and GRP are widely distributed in several peripheral tissues and throughout cod brain, although expression levels are very low. During development, ghrelin was detected at the cleavage stage, with low expression levels persisting until the first-feeding stage, while GRP was detected at the blastula stage, showing increased expression from the pre hatching stage on. Juvenile cod fed medium rations displayed periprandial changes in gut ghrelin, but not GRP, expression, with higher expression levels at meal time compared to 2h before feeding time. Ghrelin gut mRNA expression was not affected by rations, whereas GRP gut mRNA expression was higher in fish fed high rations as compared to fish fed low rations. Neither ghrelin nor GRP gut mRNA expressions were affected by 30 days starvation or 5 days re-feeding. PMID- 19121321 TI - Invitro effect of Canavalia ensiformis urease and the derived peptide Jaburetox 2Ec on Rhodnius prolixus Malpighian tubules. AB - Ureases are metalloenzymes that are widespread among plants, fungi and bacteria. Urease isoforms (jack bean urease-JBU and canatoxin) from Canavalia ensiformis seeds are toxic to insects and fungi, suggesting a role in plant defense. The entomotoxic effect is due to the release of a 10-kDa peptide by cathepsin-like enzymes in the insect's midgut. Urease causes a decrease in post-feeding weight loss in Rhodnius prolixus, suggesting an effect on water balance. To investigate how this impairment occurs, we have evaluated the action of JBU and the urease derivated peptide Jaburetox-2Ec on R. prolixus Malpighian tubules and also investigated the involvement of second messengers. JBU and Jaburetox-2Ec affect serotonin-induced secretion from Malpighian tubules. This effect is not cAMP dependent, but the Jaburetox-2Ec effect is cGMP-dependent. Eicosanoid metabolites and calcium ions appear to be involved in JBU effect on diuresis, but are not involved in the action of Jaburetox-2Ec. Jaburetox-2Ec, but not JBU, causes a change in the transepithelial potential of the tubules. Canatoxin has a similar effect on tubules secretion, decreasing the secretion rate, but the urease from Helicobacter pylori has no significant effect. These data are helpful in our understanding of the actions of ureases and derived peptides on insects, and also reinforces the potential use of these proteins as biopesticides. PMID- 19121322 TI - Theoretical investigation of action potential duration dependence on extracellular Ca2+ in human cardiomyocytes. AB - Reduction in [Ca2+]o prolongs the AP in ventricular cardiomyocytes and the QTc interval in patients. Although this phenomenon is relevant to arrhythmogenesis in the clinical setting, its mechanisms are counterintuitive and incompletely understood. To evaluate in silico the mechanisms of APD modulation by [Ca2+]o in human cardiomyocytes. We implemented the Ten Tusscher-Noble-Noble-Panfilov model of the human ventricular myocyte and modified the formulations of the rapidly and slowly activating delayed rectifier K+ currents (IKr and IKs) and L-type Ca2+ current (ICaL) to incorporate their known sensitivity to intra- or extracellular Ca2+. Simulations were run with the original and modified models at variable [Ca2+]o in the clinically relevant 1 to 3 mM range. The original model responds with APD shortening to decrease in [Ca2+]o, i.e. opposite to the experimental observations. Incorporation of Ca2+ dependency of K+ currents cannot reproduce the inverse relation between APD and [Ca2+]o. Only when ICaL inactivation process was modified, by enhancing its dependency on Ca2+, simulations predict APD prolongation at lower [Ca2+]o. Although Ca2+-dependent ICaL inactivation is the primary mechanism, secondary changes in electrogenic Ca2+ transport (by Na+/Ca2+ exchanger and plasmalemmal Ca2+-ATPase) contribute to the reversal of APD dependency on [Ca2+]o. This theoretical investigation points to Ca2+-dependent inactivation of ICaL as a mechanism primarily responsible for the dependency of APD on [Ca2+]o. The modifications implemented here make the model more suitable to analyze repolarization mechanisms when Ca2+ levels are altered. PMID- 19121323 TI - PknB-mediated phosphorylation of a novel substrate, N-acetylglucosamine-1 phosphate uridyltransferase, modulates its acetyltransferase activity. AB - Identifying direct targets of kinases and determining how their activities are regulated are central to understanding how they generate biological responses. Genetic and biochemical studies have shown that Mycobacterium tuberculosis serine/threonine protein kinases PknA and PknB play a role in modulating cell shape and possibly cell division. In this report, we show that the enzyme N acetylglucosamine-1-phosphate uridyltransferase (GlmU) of M. tuberculosis is a novel substrate of PknB and is phosphorylated on threonine residues. GlmU carries out two important biochemical activities: a C-terminal domain catalyzes the transfer of acetyl group from acetyl coenzyme A to glucosamine-1-phosphate to produce N-acetylglucosamine-1-phosphate, which is converted into UDP-N acetylglucosamine by the transfer of uridine 5'-monophosphate (from uridine 5' triphosphate), a reaction catalyzed by the N-terminal domain. We determined the crystal structures of GlmU in apo form and UDP-N-acetylglucosamine-bound form, and analyzed them to identify threonine residues that may be accessible to PknB. The structure shows a two-domain architecture, with an N-terminal domain having an alpha/beta-like fold and with a C-terminal domain that forms a left-handed parallel beta-helix structure. Kinase assays with PknB using the N- and C terminal domains of GlmU as substrates illustrated that PknB phosphorylates GlmU in the C-terminal domain. Furthermore, mutational studies reveal one of the five threonines present in region 414-439 to be phosphorylated by PknB. Structural and biochemical analyses have shown the significance of a variable C-terminal tail in regulating acetyltransferase activity. Notably, we demonstrate that although PknB mediated phosphorylation of GlmU does not affect its uridyltransferase activity, it significantly modulates the acetyltransferase activity. These findings imply a role for PknB in regulating peptidoglycan synthesis by modulating the acetyltransferase activity of GlmU. PMID- 19121324 TI - Allostery wiring diagrams in the transitions that drive the GroEL reaction cycle. AB - Determining the network of residues that transmit allosteric signals is crucial to understanding the function of biological nanomachines. During the course of a reaction cycle, biological machines in general, and Escherichia coli chaperonin GroEL in particular, undergo large-scale conformational changes in response to ligand binding. Normal mode analyses, based on structure-based coarse-grained models where each residue is represented by an alpha carbon atom, have been widely used to describe the motions encoded in the structures of proteins. Here, we propose a new Calpha-side chain elastic network model of proteins that includes information about the physical identity of each residue and accurately accounts for the side-chain topology and packing within the structure. Using the Calpha-side chain elastic network model and the structural perturbation method, which probes the response of a local perturbation at a given site at all other sites in the structure, we determine the network of key residues (allostery wiring diagram) responsible for the T-->R and R''-->T transitions in GroEL. A number of residues, both within a subunit and at the interface of two adjacent subunits, are found to be at the origin of the positive cooperativity in the ATP driven T-->R transition. Of particular note are residues G244, R58, D83, E209, and K327. Of these, R38, D83, and K327 are highly conserved. G244 is located in the apical domain at the interface between two subunits; E209 and K327 are located in the apical domain, toward the center of a subunit; R58 and D83 are equatorial domain residues. The allostery wiring diagram shows that the network of residues are interspersed throughout the structure. Residues D83, V174, E191, and D359 play a critical role in the R''-->T transition, which implies that mutations of these residues would compromise the ATPase activity. D83 and E191 are also highly conserved; D359 is moderately conserved. The negative cooperativity between the rings in the R''-->T transition is orchestrated through several interface residues within a single ring, including N10, E434, D435, and E451. Signal from the trans ring that is transmitted across the interface between the equatorial domains is responsible for the R''-->T transition. The cochaperonin GroES plays a passive role in the R''-->T transition. Remarkably, the binding affinity of GroES for GroEL is allosterically linked to GroEL residues 350-365 that span helices K and L. The movements of helices K and L alter the polarity of the cavity throughout the GroEL functional cycle and undergo large-scale motions that are anticorrelated with the other apical domain residues. The allostery wiring diagrams for the T-->R and R''-->T transitions of GroEL provide a microscopic foundation for the cooperativity (anticooperativity) within (between) the ring (rings). Using statistical coupling analysis, we extract evolutionarily linked clusters of residues in GroEL and GroES. We find that several substrate protein binding residues as well as sites related to ATPase activity belong to a single functional network in GroEL. For GroES, the mobile loop residues and GroES/GroES interface residues are linked. PMID- 19121326 TI - Limiting similarity and niche theory for structured populations. AB - We develop the theory of limiting similarity and niche for structured populations with finite number of individual states (i-state). In line with a previously published theory for unstructured populations, the niche of a species is specified by the impact and sensitivity niche vectors. They describe the population's impact on and sensitivity towards the variables involved in the population regulation. Robust coexistence requires sufficient segregation of the impact, as well as of the sensitivity niche vectors. Connection between the population-level impact and sensitivity and the impact/sensitivity of the specific i-states is developed. Each i-state contributes to the impact of the population proportional to its frequency in the population. Sensitivity of the population is composed of the sensitivity of the rates of demographic transitions, weighted by the frequency and by the reproductive value of the initial and final i-states of the transition, respectively. Coexistence in a multi-patch environment is studied. This analysis is interpreted as spatial niche segregation. PMID- 19121325 TI - Functional analysis of the transmembrane domain in paramyxovirus F protein mediated membrane fusion. AB - To enter cells, enveloped viruses use fusion-mediating glycoproteins to facilitate the merger of the viral and host cell membranes. These glycoproteins undergo large-scale irreversible refolding during membrane fusion. The paramyxovirus parainfluenza virus 5 mediates membrane merger through its fusion protein (F). The transmembrane (TM) domains of viral fusion proteins are typically required for fusion. The TM domain of F is particularly interesting in that it is potentially unusually long; multiple calculations suggest a TM helix length between 25 and 48 residues. Oxidative cross-linking of single-cysteine substitutions indicates the F TM trimer forms a helical bundle within the membrane. To assess the functional role of the paramyxovirus parainfluenza virus 5 F protein TM domain, alanine scanning mutagenesis was performed. Two residues located in the outer leaflet of the bilayer are critical for fusion. Multiple amino acid substitutions at these positions indicate the physical properties of the side chain play a critical role in supporting or blocking fusion. Analysis of intermediate steps in F protein refolding indicated that the mutants were not trapped at the open stalk intermediate or the prehairpin intermediate. Incorporation of a known F protein destabilizing mutation that causes a hyperfusogenic phenotype restored fusion activity to the mutants. Further, altering the curvature of the lipid bilayer by addition of oleic acid promoted fusion of the F protein mutants. In aggregate, these data indicate that the TM domain plays a functional role in fusion beyond merely anchoring the protein in the viral envelope and that it can affect the structures and steady-state concentrations of the various conformational intermediates en route to the final postfusion state. We suggest that the unusual length of this TM helix might allow it to serve as a template for formation of or specifically stabilize the lipid stalk intermediate in fusion. PMID- 19121328 TI - Are hemianopic reading and visual exploration impairments visually elicited? New insights from eye movements in simulated hemianopia. AB - Hemianopic reading and visual exploration impairments are well-known clinical phenomena. Yet, it is unclear whether they are primarily caused by the hemianopic visual field defect itself or by additional brain injury preventing efficient spontaneous oculomotor adaptation. To establish the extent to which these impairments are visually elicited we simulated unilateral homonymous hemianopia in healthy participants, using a gaze-contingent display paradigm, and investigated its effect on reading and visual exploration. We demonstrate that simulated hemianopia induces the reading and visual exploration impairments of hemianopic patients. Over time, however, all participants showed efficient spontaneous oculomotor adaptation to the visual-sensory loss which improved their reading and visual exploration performance. Our results suggest that the hemianopic visual field defect is a major component of the chronic impairments of reading and visual and exploration found in hemianopic patients although it may not be their sole cause. PMID- 19121329 TI - Dexamethasone downregulates the expression of parathyroid hormone-related protein (PTHrP) in mesenchymal stem cells. AB - Parathyroid hormone-related protein (PTHrP) has been shown to have anabolic effects in women with postmenopausal osteoporosis. PTHrP promotes the recruitment of osteogenic cells and prevents apoptotic death of osteoblasts and osteocytes. The receptor responsible for the effects of PTHrP is the common PTH/PTHrP receptor (PTH1R). Glucocorticoids (GC) are commonly used as drugs to treat inflammatory diseases. Long-term GC treatments are often associated with bone loss which can lead to GC-induced osteoporosis. The aim of this work was to study the effects of the glucocorticoid dexamethasone (Dex) on the expression of PTHrP and PTH1R in adult human mesenchymal stem cells, the progenitor cells of osteoblasts. Adult human mesenchymal stem cells (hMSC) were cultured and differentiated by standard methods. The expression of PTHrP and PTH1R mRNA was assayed by real-time qPCR. The PTHrP release into the culture media was measured by an immunoradiometric assay. Treatment with Dex (10 nM) resulted in an 80% drop in the PTHrP release within 6 h. A 24 h Dex treatment also reduced the expression of PTHrP mRNA by up to 90%. The expression of PTH1R receptor mRNA was simultaneously increased up to 20-fold by 10 nM Dex. The effects of Dex on PTHrP and PTH1R were dose-dependent and experiments with the GC-receptor antagonist mifepristone showed an involvement of GC-receptors in these effects. In addition to the Dex-induced effects on PTHrP and PTH1R, Dex also increased mineralization and the expression of the osteoblast markers Runx2 and alkaline phosphatase. In our studies, we show that dexamethasone decreases the expression of PTHrP and increases the expression of the PTH1R receptor. This could have an impact on PTHrP-mediated anabolic actions on bone and could also affect the responsiveness of circulating PTH. The results indicate that glucocorticoids affect the signalling pathway of PTHrP by regulating both PTHrP and PTH1R expression and these mechanisms could be involved in glucocorticoid-induced osteoporosis. PMID- 19121327 TI - Protective effects of high-molecular weight polyethylene glycol (PEG) in human lung endothelial cell barrier regulation: role of actin cytoskeletal rearrangement. AB - Acute lung injury represents the result of multiple pathways initiated by local or systemic insults and is characterized by profound vascular permeability, pulmonary edema, and life-threatening respiratory failure. Permeability-reducing therapies are of potential clinical utility but are currently unavailable. We hypothesized that polyethylene glycol (PEG) compounds, inert and non-toxic polymers that serve as a surrogate mucin lining in intestinal epithelium, may attenuate agonist-mediated lung endothelial cell (EC) barrier dysfunction. High molecular weight PEG (PEG15-20) produced rapid, dose-dependent increases in transendothelial electrical resistance (TER) in human lung endothelium cultured on gold microelectrodes, reflecting increased paracellular integrity. The maximal effective concentration of 8% PEG induced a sustained 125% increase in TER (40 h), results similar to barrier-enhancing agonists such as sphingosine 1-phosphate (40% increase in TER). Maximal PEG barrier enhancement was achieved at 45-60 min and PEG effectively reversed both thrombin- and LPS-induced EC barrier dysfunction. Consistent with the increase in TER, immunofluorescent studies demonstrated that PEG produced significant cytoskeletal rearrangement with formation of well-defined cortical actin rings and lamellipodia containing the actin-binding proteins, cortactin and MLCK, known participants in cell-matrix and cell-cell junctional adhesion. Finally, PEG challenge induced rapid alterations in levels of MAP kinase and MLC phosphorylation. In summary, PEG joins a number of EC barrier-regulatory agents which rapidly activate barrier-enhancing signal transduction pathways which target the cytoskeleton and provides a potential therapeutic strategy in inflammatory lung injury. PMID- 19121330 TI - The hyperthermia mediated by 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) is sensitive to sex differences. AB - Female subjects have been reported to be less sensitive to the hyperthermic effects of 3,4-methylenedioxymethamine (MDMA) than males. Studies were designed to examine the cellular mechanisms involved in these sex sensitive differences. Gonadectomized female and male rats were treated with a 200 microg 100 microL(-1) of estrogen or 100 microg 100 microL(-1) of testosterone respectively every 5 days for a total of three doses. Rats were then challenged with either saline or MDMA (20 mg kg(-1), sc). Rats were then euthanized and aortas were constricted, in vitro, by serial phenylephrine (Phe) addition with or without the inhibitor of nitric oxide (NO) synthase, g-nitro-L-Arginine-Methyl Ester (L-NAME). Skeletal muscle uncoupling protein-3 (UCP3) expression was measured as well as plasma norepinephrine (NE) levels. All males but no females developed hyperthermia following MDMA treatment. The EC(50) for Phe dose response curves increased only in the females treated with MDMA and T(max) for Phe increased following L-NAME only in the females. Both males and females demonstrated an increase in plasma NE following MDMA treatment; however, males displayed a significantly greater NE concentration. Skeletal muscle UCP3 expression was 80% less in females than in males. These results suggest that the inability of MDMA to induce a thermogenic response in the female subjects may be due to four sex-specific mechanisms: 1) Female subjects have reduced sympathetic activation following MDMA challenge; 2) Female vasculature is less sensitive to alpha(1)-AR stimulation following MDMA challenge; 3) Female vasculature has an increased sensitivity to NO; 4) UCP3 expression in skeletal muscle is less in females. PMID- 19121331 TI - Acute nicotine exposure and modulation of a spinal motor circuit in embryonic zebrafish. AB - The zebrafish model system is ideal for studying nervous system development. Ultimately, one would like to link the developmental biology to various aspects of behavior. We are studying the consequences of nicotine exposure on nervous system development in zebrafish and have previously shown that chronic nicotine exposure produces paralysis. We also have made observations that the embryos moved in the initial minutes of the exposure as the bend rates of the musculature increased. This nicotine induced behavior manifests as an increase in the rate of spinal musculature bends, which spontaneously begin at approximately 17 h post fertilization. The behavioral observations prompted the systematic characterization of nicotine-induced modulation of zebrafish embryonic motor output; bends of the trunk musculature. We first characterized embryonic motor output in zebrafish embryos with and without their chorions. We then characterized the motor output in embryos raised at 28 degrees C and 25 degrees C. The act of dechorionation along with temperature influenced the embryonic bend rate. We show that nicotine exposure increases embryonic motor output. Nicotine exposure caused the musculature bends to alternate in a left-right-left fashion. Nicotine was able to produce this phenotype in embryos lacking supraspinal input. We then characterize the kinetics of nicotine influx and efflux and demonstrate that nicotine as low as 1 microM can disrupt embryonic physiology. Taken together, these results indicate the presence of nicotinic acetylcholine receptors (nAChRs) associated with embryonic spinal motor circuits early in embryogenesis. PMID- 19121332 TI - Regulation of gap junctional intercellular communication by TCDD in HMEC and MCF 7 breast cancer cells. AB - Previous studies suggest that many neoplastic tissues exhibit a decrease in gap junctional intercellular communication (GJIC). Many hydrocarbons and organochlorine compounds are environmental pollutants known to be carcinogenic. The effect of an organochlorine compound, TCDD, on GJIC in human breast cell lines has not been established. In the present study, we showed that TCDD causes an inhibition in the gap junctional activity in MCF-7 (breast cancer cells). In MCF-7 cells, an increase in the phosphorylated form of gap junctional protein, connexin 43 (Cx43), and PKC alpha was seen in the presence of TCDD. Gap junctional plaque formation was significantly decreased in MCF-7 cells in the presence of TCDD. Immunoprecipitation studies of PKC alpha showed that TCDD caused a significant 40% increase in the phosphorylated Cx43 in MCF-7 cells. TCDD also modulated the translocation of PKC alpha from the cytosol to the membrane and caused a 2-fold increase in the PKC alpha activity at 50 nM TCDD in MCF-7 cells. Calphostin C, an inhibitor of PKC alpha, showed a significant inhibition of PKC alpha activity in the presence of TCDD. Furthermore, TCDD also caused a decrease in the gap junctional activity and Cx43 protein in human mammary epithelial cells (HMEC). However, we observed a shift in the Cx43 plaques towards the perinuclear membrane in the presence of TCDD by confocal microscopy and Western blot. Overall, these results conclude that TCDD decreases GJIC by phosphorylating Cx43 via PKC alpha signaling pathway in MCF-7 cells; however, TCDD decreases the GJIC by affecting the localization of Cx43 in HMEC. These new findings elucidate the differential mode of effect of TCDD in the downregulation of GJIC in HMEC and MCF-7 cells. PMID- 19121333 TI - Chronic low-level arsenic exposure causes gender-specific alterations in locomotor activity, dopaminergic systems, and thioredoxin expression in mice. AB - Arsenic (As) is a toxic metalloid widely present in the environment. Human exposure to As has been associated with the development of skin and internal organ cancers and cardiovascular disorders, among other diseases. A few studies report decreases in intelligence quotient (IQ), and sensory and motor alterations after chronic As exposure in humans. On the other hand, studies of rodents exposed to high doses of As have found alterations in locomotor activity, brain neurochemistry, behavioral tasks, and oxidative stress. In the present study both male and female C57Bl/6J mice were exposed to environmentally relevant doses of As such as 0.05, 0.5, 5.0, or 50 mg As/L of drinking water for 4 months, and locomotor activity was assessed every month. Male mice presented hyperactivity in the group exposed to 0.5 mg As/L and hypoactivity in the group exposed to 50 mg As/L after 4 months of As exposure, whereas female mice exposed to 0.05, 0.5, and 5.0 mg As/L exhibited hyperactivity in every monthly test during As exposure. Furthermore, striatal and hypothalamic dopamine content was decreased only in female mice. Also decreases in tyrosine hydroxylase (TH) and cytosolic thioredoxin (Trx-1) mRNA expression in striatum and nucleus accumbens were observed in male and female mice, respectively. These results indicate that chronic As exposure leads to gender-dependent alterations in dopaminergic markers and spontaneous locomotor activity, and down-regulation of the antioxidant capacity of the brain. PMID- 19121335 TI - Thermal aggregation of a model allosteric protein in different conformational states. AB - Protein aggregation is of crucial importance in a wide variety of situations. High temperatures, combined with other denaturing conditions, have been used very extensively to decipher some of the fundamentals related to formation of amorphous and fibrillar protein aggregates. The present study reports on the dependency of thermal aggregation of bovine liver glutamate dehydrogenase (GDH), a well-characterized allosteric enzyme, on its conformational state. The initial phases of thermal aggregation of this protein was followed in the presence of a number of well-known allosteric ligands. Positive effectors were found to decrease the rate and extent of aggregation in a concentration dependent manner, while negative effectors did the reverse. ADP, one of the most characterized GDH activators was found to stabilize a specific protein conformation resulting in loss of propensity to aggregate. The importance of this observation related to control of protein-protein interactions leading to protein aggregation is discussed. PMID- 19121334 TI - Rodent models of nicotine reward: what do they tell us about tobacco abuse in humans? AB - Tobacco products are widely abused in humans, and it is assumed that nicotine is the key substrate in these products that produces addiction. Based on this assumption, several pre-clinical studies have utilized animal models to measure various aspects of nicotine addiction. Most of this work has focused on behavioral measures of nicotine and how other variables contribute to these effects. Here we discuss the most commonly used animal models including, self administration (SA), place conditioning (PC), and the intracranial self stimulation (ICSS) paradigms in rodents. The strengths, limitations and procedural variables of these models are reviewed, followed by a discussion of how the animal models have been used to study factors such as age, sex, stress, and the effects of tobacco products other than nicotine. These factors are discussed in light of their influences on human tobacco abuse. The rodent models are evaluated in the context of face, predictive, and construct validity, and we propose that inclusion of factors such as age, sex, stress and other constituents of tobacco aside from nicotine can increase the utility of these animal models by more closely mimicking human tobacco abuse. PMID- 19121337 TI - Dendritic cells from bench to bedside and back. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells of the immune system. They serve as the sentinels that capture antigens in the periphery, process them into peptides and present these to lymphocytes in lymph nodes. DCs play a key role in regulating immunity. Several DC-subsets exist, including myeloid-DCs (MDCs), plasmacytoid-DCs (PDCs) and Langerhans cells (LC). DCs not only instruct T- and B-lymphocytes, but also activate Natural Killer cells and produce interferons, thus linking the innate and adaptive immune system. Inflammatory-mediators and especially the Toll like receptor (TLR) family of proteins have been shown to play a pivotal role in inducing the immune activation program in DCs. TLRs recognize pathogen-associated-molecular-patterns (PAMPS) like LPS or flagellin and signal to alert immune cells in general, and DC in particular. DC activation, also referred to as DC maturation, thus results in immunity. In contrast, resting DC or DC receiving immune-inhibitory signals, like IL-10 and/or corticosteroids, induce immune tolerance via T cell deletion and induction of suppressive T cells, now termed regulatory T cells. Several mouse models have demonstrated that the immunological outcome is depending on the DC activation state; mature immune-activating DC protect mice from a tumor or pathogen while tolerogenic DC induces tolerance against transplanted tissues. Hence, DC acts at the interface of immunity and peripheral tolerance. PMID- 19121336 TI - Optical coherence tomography (OCT) reveals depth-resolved dynamics during functional brain activation. AB - Optical intrinsic signal imaging (OISI) provides two-dimensional, depth integrated activation maps of brain activity. Optical coherence tomography (OCT) provides depth-resolved, cross-sectional images of functional brain activation. Co-registered OCT and OISI imaging was performed simultaneously on the rat somatosensory cortex through a thinned skull during forepaw electrical stimulation. Fractional signal change measurements made by OCT revealed a functional signal that correlates well with that of the intrinsic hemodynamic signals and provides depth-resolved, layer-specific dynamics in the functional activation patterns indicating retrograde vessel dilation. OCT is a promising a new technology which provides complementary information to OISI for functional neuroimaging. PMID- 19121339 TI - Flagellar localization of a novel isoform of myosin, myosin XXI, in Leishmania. AB - Leishmania major genome analysis revealed the presence of putative genes corresponding to two myosins, which have been designated to class IB and a novel class, class XXI, specifically present in kinetoplastids. To characterize these myosin homologs in Leishmania, we have cloned and over-expressed the full-length myosin XXI gene and variable region of myosin IB gene in bacteria, purified the corresponding proteins, and then used the affinity purified anti-sera to analyze the expression and intracellular distribution of these proteins. Whereas myosin XXI was expressed in both the promastigote and amastigote stages, no expression of myosin IB could be detected in any of the two stages of these parasites. Further, myosin XXI expression was more predominant in the promastigote stage where it was preferentially localized in the proximal region of the flagellum. The observed flagellar localization was not dependent on the myosin head region or actin but was exclusively determined by the myosin tail region, as judged by over-expressing GFP conjugates of full-length myosin XXI, its head domain and its tail domain separately in Leishmania. Furthermore, immunofluorescence and immuno gold electron microscopy analyses revealed that this protein was partly associated with paraflagellar rod proteins but not with tubulins in the flagellar axoneme. Our results, for the first time, report the expression and detailed analysis of cellular localization of a novel class of myosin, myosin XXI in trypanosomatids. PMID- 19121338 TI - Does 5-bromo-2'-deoxyuridine (BrdU) disrupt cell proliferation and neuronal maturation in the adult rat hippocampus in vivo? AB - 5-Bromo-2'-deoxyuridine (BrdU) is frequently used as a mitotic marker in studies of cell proliferation. Recent studies have reported cytotoxic effects of BrdU on neural progenitor cells in embryonic and neonatal brains in vivo and in adult tissue studied in vitro. The present study was conducted to assess whether BrdU interferes with cell proliferation and neuronal maturation in the rat adult hippocampus in vivo. BrdU effects across a wide range of doses (40-480 mg/kg) on cell proliferation and the population of immature neurons in the adult hippocampus were investigated using immunohistochemical labeling methods for the cell cycle marker Ki67 and a marker for immature neurons, doublecortin. BrdU did not influence cell proliferation in the dentate gyrus or the population of immature neurons observed in the adult hippocampus relative to those observed in saline treated controls. Thus, in contrast with reports of deleterious effects of BrdU in embryonic and neonatal tissue and adult tissue studied in vitro, BrdU does not appear to have cytotoxic effects on proliferating hippocampal cells or immature neurons in vivo in rats. PMID- 19121340 TI - Bcl-XL mediates epidermal growth factor dependent cell survival in HC11 mammary epithelial cells. AB - Apoptosis is the predominant process controlling cell deletion during post lactational mammary gland remodeling. The members of the Bcl-2 protein family, whose expression levels are under the control of lactogenic hormones, internally control this mechanism. Epidermal growth factor (EGF) belongs to a family of proteins that act as survival factors for mammary epithelial cells upon binding to specific membrane tyrosine kinase receptors. Expression of EGF peaks during lactation and dramatically decreases in the involuting mammary gland. Though it was suggested that the protective effect of EGF is mediated through the phosphatidylinositol-3-kinase (PI3K) or MEK/ERK kinases activities, little is known about the downstream mechanisms involved on the anti-apoptotic effect of EGF on mammary epithelial cells; particularly the identity of target genes controlling apoptosis. Here, we focused on the effect of EGF on the survival of mammary epithelial cells. We particularly aimed at the characterization of the signaling pathways that were triggered by this growth factor, impinge upon expression of Bcl-2 family members and therefore have an impact on the regulation of cell survival. We demonstrate that EGF provokes the induction of the anti apoptotic isoform Bcl-XL and the phosphorylation and down-regulation of the pro apoptotic protein Bad. The activation of JNK and PI3K/AKT signaling pathways promotes the induction of Bcl-XL while AKT activation also leads to Bad phosphorylation and down-regulation. This protective effect of EGF correlates mainly with the up-regulation of Bcl-XL than with the down-regulation of Bad. In fact, HC11 cells unable to express bcl-X, die even in the presence of EGF. In this context, Bcl-XL emerges as a key anti-apoptotic molecule critical for mediating EGF cell survival. PMID- 19121341 TI - Binding of S100 proteins to RAGE: an update. AB - The Receptor for Advanced Glycation Endproducts (RAGE) is a multi-ligand receptor of the immunoglobulin family. RAGE interacts with structurally different ligands probably through the oligomerization of the receptor on the cell surface. However, the exact mechanism is unknown. Among RAGE ligands are members of the S100 protein family. S100 proteins are small calcium binding proteins with high structural homology. Several members of the family have been shown to interact with RAGE in vitro or in cell-based assays. Interestingly, many RAGE ligands appear to interact with distinct domains of the extracellular portion of RAGE and to trigger various cellular effects. In this review, we summarize the modes of S100 protein-RAGE interaction with regard to their cellular functions. PMID- 19121342 TI - Aggression and 5HTT polymorphism in females: study of synchronized swimming and control groups. AB - Aggression is a heterogeneous heritable psychological trait, also influenced by environmental factors. Previous studies, mostly conducted on male population, have found some associations of the aggression with the polymorphisms of genes, regulating the activity of serotonin (5-HT) in the brain. However, psychological as well as biochemical manifestations of the aggression are different in males and females. Our study aimed to investigate the association of 5-HTT gene polymorphism with different facets of aggression (BDHI) in females. Two groups: the synchronized swimming and non-athlete control, - were examined to study the possible modulation effect of sport on the association between 5-HTT gene polymorphism and aggression. It was found that in both groups the low-active 5 HTT polymorphism (SS) was associated with increased scores on Indirect Hostility scale and decreased scores on Negativism scale, compared to LL genotype. No interaction effect between sport and 5-HTT polymorphism was found. The higher percentage of LL-carriers and lower of LS-carriers in the synchronized swimming group compared to the control one was observed. This may be the sign of the importance of LL polymorphism of 5-HTT gene, previously associated with higher resistance to stress factors, for being an athlete, although this result has to be taken cautiously keeping in mind the stratification problem. Synchronized swimmers had lower scores on Assault, Negativism, Irritability and Verbal Hostility compared to age-matched control girls (in general and for each 5-HTT genotype separately), suggesting that they may have more matured emotional system (older control group has also lower scores on these scales). PMID- 19121343 TI - Marked increases in hippocampal neuron indoleamine 2, 3-dioxygenase via IFN-gamma independent pathway following transient global ischemia in mouse. AB - Indoleamine 2, 3-dioxygenase (IDO), which catabolizes L-tryptophan (L-TRP) to L kynurenine (L-KYN), is an immunoregulatory factor that is up-regulated via an interferon-gamma (IFN-gamma)-dependent and/or -independent mechanism. In this study, we investigated the localization of IDO and whether induction of IDO expression is an IFN-gamma-dependent and/or -independent mechanism in the CNS after cerebral ischemia. The expressions of IDO protein and mRNA were investigated at different time points following cerebral ischemia using immunohistochemistry, immunofluorescence and RT-PCR. Hippocampal neuron IDO mRNA and immunohistochemical staining were significantly up-regulated 72h after transient global ischemia. Although IFN-gamma is a dominant inducer of IDO, hippocampal neuron IDO was clearly up-regulated in IFN-gamma KO mice. In summary, this is the first finding that up-regulation of IDO in hippocampal neurons after transient global ischemia occurs via INF-gamma-independent mechanisms. PMID- 19121344 TI - Metabolic engineering of the L-valine biosynthesis pathway in Corynebacterium glutamicum using promoter activity modulation. AB - The previously constructed strain Corynebacterium glutamicumilvNM13 with acetohydroxy acid synthase, resistant to inhibition by all three branched-chain amino acids (L-valine, L-isoleucine and L-leucine), was used as a basis to develop a new type of valine producer by genetic engineering. The main strategy was to modulate expression of the genes involved in the biosynthesis of branched chain amino acids. The activity of the promoters P-ilvD (dihydroxyacid dehydratase) and P-ilvE (transaminase) was up-modulated and the activity of the promoters P-ilvA (threonine deaminase) and P-leuA (isopropylmalate synthase) was down-modulated by site-directed mutagenesis. A constructed weak promoter of ilvA (or leuA), which was introduced into the C. glutamicum chromosome via a gene replacement technique reduced the biosynthetic rate of isoleucine (or leucine), which lowered the mutant growth rate and increased valine production. Overexpression of ilvD and ilvE driven by the strong mutant promoters P-ilvDM7 and P-ilvEM6 resulted in an even higher level of valine production. Thus, the strain C. glutamicum ilvNM13 DeltapanB P-ilvAM1CG P-ilvDM7 P-ilvEM6, having all mutations constructed within the chromosome, produced 136 mM valine in a 48-h cultivation. PMID- 19121345 TI - Development of a polysulfone hollow fiber vascular bio-artificial pancreas device for in vitro studies. AB - A system for vascular hollow fiber bio-artificial pancreas development, optimization and in vitro testing was implemented and operated in a simple and fully described manner, allowing other researchers to test a variety of experimental conditions (different biomaterials, biologic tissue, addition of proteins or other adjuvants). In this work, a polysulfone hollow fiber was used as bioprotective material. Two different cell sources were co-immobilized with agarose microspheres in and experimented with the membrane device: rat islets of Langerhans and mouse beta-TC-3 insulinoma cells. The results obtained with islets of Langerhans were used as islet comparable insulin-release data. Beta-TC-3 cells were mainly used in these studies, due to higher control and reproducibility of cell number and behavior: addition of hemoglobin was beneficial for sustained cell viability, especially during cell insertion in the device (viability assessed by beta-TC-3 lactate dehydrogenase activity in the recirculating culture medium); cells did not adhere to the polysulfone membrane (assessed by SEM observation of membrane samples from dynamic cultures). Comparable device functionality and insulin-release results were attained with both cell types: device functionality was maintained for 7-9 days and maximum insulin-release during dynamic glucose challenges were 2.6 x 10(-3)+/-7 x 10(-5)microU/beta-cell x 8 h, with islets, and 9.3 x 10(-4)+/-2 x 10(-5)microU/beta-cell x 2 h, with beta-TC-3 cells. PMID- 19121346 TI - Identification and characterization of a native epitope common to norovirus strains GII/4, GII/7 and GII/8. AB - Norovirus is an important cause of acute non-bacterial gastroenteritis in humans. The norovirus genus is comprised of at least five genogroups based on sequence differences. The norovirus genogroup II (GII/4) strain is recognized as the predominant genotype worldwide. We expressed a 60 kDa full-length recombinant capsid protein of norovirus GII/4 in Escherichia coli and generated three monoclonal antibodies (MAbs) against it. Western blotting indicated that all three MAbs had reactivity against the recombinant capsid protein and a 58 kDa native capsid protein of norovirus obtained from stool samples. MAb-capture ELISA showed that MAb detected segmental strains within GII antigens in clinical material. To identify the existent range of this epitope, epitope analyses were processed by expressing 12 amino acids of the GST-fusion peptides. The epitope analyses revealed that the MAb N2C3 recognized a continuous native epitope (55)WIRNNF(60) in the shell domain, which not only belongs to strain GII/4, but also to strains GII/7 and GII/8. This is a new native epitope to be reported for norovirus GII/4. PMID- 19121347 TI - Lipid extrudates as novel sustained release systems for pharmaceutical proteins. AB - In order to develop improved lipid-based implants for proteins, the applicability of twin screw extrusion as a manufacturing strategy was investigated. Using lipid blends of low and high melting lipids, extrusion could be performed at moderate temperatures. In addition to the lipids, the implant systems contained 10% rh interferon alpha-2a (IFN-alpha) co-lyophilised with hydroxypropyl-beta cyclodextrin (HP-beta-CD), and 10% or 20% polyethylene glycol (PEG), respectively. Protein integrity was analysed by SDS-PAGE after extraction. FTIR spectroscopy was used as a non-invasive method to assess the secondary protein structure of IFN-alpha within the lipid extrudates. Both methods indicate the preservation of the protein structure. Depending on the diameter of the prepared extrudates and on the PEG content of the system, protein release occurred in a sustained manner over 15, 40, or 60 days. Interestingly, extrudates with a smaller diameter revealed a continuous release profile, whereas extrudates with a diameter of 1.9 mm delivered IFN-alpha in a triphasic profile with a burst, lag and linear release phase lasting over 13 or 26 days. In conclusion, lipid implants for proteins can be prepared by twin screw extrusion in an elegant and controllable manner that allows the adjustment of release characteristics. PMID- 19121348 TI - An injectable hyaluronic acid-tyramine hydrogel system for protein delivery. AB - Previously, we reported the independent tuning of mechanical strength (crosslinking density) and gelation rate of an injectable hydrogel system composed of hyaluronic acid-tyramine (HA-Tyr) conjugates. The hydrogels were formed through the oxidative coupling of tyramines which was catalyzed by hydrogen peroxide (H(2)O(2)) and horseradish peroxidase (HRP). Herein, we studied the encapsulation and release of model proteins using the HA-Tyr hydrogel. It was shown that the rapid gelation achieved by an optimal concentration of HRP could effectively encapsulate the proteins within the hydrogel network and thus prevented the undesired leakage of proteins into the surrounding tissues after injection. Hydrogels with different mechanical strengths were formed by changing the concentration of H(2)O(2) while maintaining the rapid gelation rate. The mechanical strength of the hydrogel controlled the release rate of proteins: stiff hydrogels released proteins slower compared to weak hydrogels. In phosphate buffer saline, alpha-amylase (negatively charged) was released sustainably from the hydrogel. Conversely, the release of lysozyme (positively charged) discontinued after the fourth hour due to electrostatic interactions with HA. In the presence of hyaluronidase, lysozymes were released continuously and completely from the hydrogel due to degradation of the hydrogel network. The activities of the released proteins were mostly retained which suggested that the HA-Tyr hydrogel is a suitable injectable and biodegradable system for the delivery of therapeutic proteins. PMID- 19121349 TI - One-step preparation of rifampicin/poly(lactic-co-glycolic acid) nanoparticle containing mannitol microspheres using a four-fluid nozzle spray drier for inhalation therapy of tuberculosis. AB - We prepared rifampicin (RFP)/poly(lactic-co-glycolic acid) (PLGA) nanoparticle containing mannitol (MAN) microspheres for inhalation therapy of tuberculosis using a four-fluid nozzle spray drier in one step. The RFP and PLGA acetone/methanol (2/1) solution and MAN aqueous solution were supplied through different liquid passages of the four-fluid nozzle spray drier to obtain MAN microspheres including RFP/PLGA nanoparticles. The mean diameter of RFP/PLGA nanoparticles dispersed in MAN was 213 nm. The (RFP/PLGA)/MAN microspheres showed good in vitro aerosol performance evaluated using a cascade impactor. In an in vitro study of uptake by alveolar macrophage cells evaluated using coumarin 6 as a marker, the PLGA microparticles were more efficiently taken in than the PLGA nanoparticles. In an in vivo study of uptake of RFP by alveolar macrophages in the lungs of rats, the uptake of RFP from the RFP/PLGA microspheres was very small. The uptake of RFP from the (RFP/PLGA)/MAN microspheres, however, was larger, about 4%, at 1 h after administration increasing to 9.3% at 4 h. An in vivo imaging study suggests that the micron-sized PLGA particles were rapidly excreted from the lungs, but that it may be difficult to clear the nano-sized PLGA particles, resulting in their retention. Thus, the RFP/PLGA nanoparticles were recognized by alveolar macrophages and the uptake of RFP increased. This study shows that it is feasible to prepare RFP/PLGA nanoparticle-containing microspheres using a four-fluid nozzle spray drier in one step, and that the particles are retained in the lungs for prolonged periods and targeted to alveolar macrophages. PMID- 19121350 TI - Nanoparticles for nasal vaccination. AB - The great interest in mucosal vaccine delivery arises from the fact that mucosal surfaces represent the major site of entry for many pathogens. Among other mucosal sites, nasal delivery is especially attractive for immunization, as the nasal epithelium is characterized by relatively high permeability, low enzymatic activity and by the presence of an important number of immunocompetent cells. In addition to these advantageous characteristics, the nasal route could offer simplified and more cost-effective protocols for vaccination with improved patient compliance. The use of nanocarriers provides a suitable way for the nasal delivery of antigenic molecules. Besides improved protection and facilitated transport of the antigen, nanoparticulate delivery systems could also provide more effective antigen recognition by immune cells. These represent key factors in the optimal processing and presentation of the antigen, and therefore in the subsequent development of a suitable immune response. In this sense, the design of optimized vaccine nanocarriers offers a promising way for nasal mucosal vaccination. PMID- 19121351 TI - Preliminary validation of the Yale Food Addiction Scale. AB - Previous research has found similarities between addiction to psychoactive substances and excessive food consumption. Further exploration is needed to evaluate the concept of "food addiction," as there is currently a lack of psychometrically validated measurement tools in this area. The current study represents a preliminary exploration of the Yale Food Addiction Scale (YFAS), designed to identify those exhibiting signs of addiction towards certain types of foods (e.g., high fat and high sugar). Survey data were collected from 353 respondents from a stratified random sample of young adults. In addition to the YFAS, the survey assessed eating pathology, alcohol consumption and other health behaviors. The YFAS exhibited adequate internal reliability, and showed good convergent validity with measures of similar constructs and good discriminant validity relative to related but dissimilar constructs. Additionally, the YFAS predicted binge-eating behavior above and beyond existing measures of eating pathology, demonstrating incremental validity. The YFAS is a sound tool for identifying eating patterns that are similar to behaviors seen in classic areas of addiction. Further evaluation of the scale is needed, especially due to a low response rate of 24.5% and a non-clinical sample, but confirmation of the reliability and validity of the scale has the potential to facilitate empirical research on the concept of "food addiction". PMID- 19121352 TI - Cloning of neuropeptide-like precursor 1 in the gray flesh fly and peptide identification and expression. AB - The neuropeptide-like precursor 1 (NPLP1) was first identified in a peptidomics experiment on Drosophila melanogaster. Limited data on this novel neuropeptide precursor suggest a role in the regulation of ecdysis in holometabolous larvae. In this study, we characterized the NPLP1 precursor in the gray flesh fly, Neobellieria bullata, which is an excellent model for physiological assays and hence to discover the role of the NPLP1 peptides. Antisera against three of the D. melanogaster NPLP1 neuropeptides stained an identical set of neurons in the central nervous system of N. bullata compared to D. melanogaster. A novel approach was applied to identify the N. bullata NPLP1 orthologs. Using a combination of affinity chromatography, mass spectrometry, cDNA cloning and RACE experiments, we obtained almost the complete coding sequence of the NPLP1 mRNA. Three encoded NPLP1 peptides were identified in central nervous system extracts by mass spectrometry. Neither doses of 25-250pmol of synthetic Neb-MGYamide and Neb-PQNamide peptides, nor the NPLP1 antisera did affect the speed of retraction, contraction and tanning in the pupariation bioassay. PMID- 19121353 TI - Clock genes regulate neurogenic transcription factors, including NeuroD1, and the neuronal differentiation of adult neural stem/progenitor cells. AB - The circadian clock system plays multiple roles in our bodies, and clock genes are expressed in various brain regions, including the lateral subventricular zone (SVZ) where neural stem/progenitor cells (NSPCs) persist and postnatal neurogenesis continues. However, the functions of clock genes in adult NSPCs are not well understood. Here, we first investigated the expression patterns of Clock and Bmal1 in the SVZ by immunohistochemistry and then verified how the expression levels of 17 clock and clock-related genes changed during differentiation of cultured adult NSPCs using quantitative RT-PCR. Finally, we used RNAi to observe the effects of Clock and Bmal1 on neuronal differentiation. Our results revealed that Clock and Bmal1 were expressed in the SVZ and double-stained with the neural progenitor marker Nestin and neural stem marker GFAP. In cultured adult NSPCs, the clock genes changed their expression patterns during differentiation, and interestingly, Bmal1 started endogenous oscillation. Moreover, gene silencing of Clock or Bmal1 by RNAi decreased the percentages of neuronal marker Map2-positive cells and expression levels of NeuroD1 mRNA. These findings suggest that clock genes are involved in the neuronal differentiation of adult NSPCs and may extend our understanding of various neurological/psychological disorders linked to adult neurogenesis and circadian rhythm. PMID- 19121354 TI - NAC1, a POZ/BTB protein that functions as a corepressor. AB - We now demonstrate that NAC1 acts as a corepressor for other POZ/BTB proteins. NAC1 is a POZ/BTB motif containing transcriptional repressor protein. In a mammalian two hybrid assay in neuronal (N2A) cells and non-neuronal (HEK 293T) cells, VP16 activation domain tagged NAC1 resulted in significant reversal of transcriptional inhibition with the Gal4-ZID, Gal4-BCL6, Gal4-ZF5, and kelch proteins Gal4-MAYVEN and Gal4-NRP/B fusion proteins. We also observed similar results with another corepressor, BCoR Gal4 fusion protein. NAC1 potentiated ZF5 mediated repression in Gal4-DBD fusion transient assays. GST pulldown assays further confirmed protein-protein interactions between these proteins and NAC1. Both the NAC1 isoforms demonstrated selective interaction through the POZ/BTB domain but not with the non-POZ/BTB region. Endogenous NAC1 and BCL6 physically associated in CNS regions. Strikingly, NAC1 did not interact with the pro myelocytic leukemia zinc finger protein (PLZF), another POZ/BTB protein that is not found in the adult brain. Therefore, we conclude that NAC1 functions as a corepressor for POZ/BTB proteins expressed in the mature CNS. PMID- 19121355 TI - 3D-pharmacophore model for RXR(gamma) agonists. AB - Three-dimensional pharmacophore models were generated for retinoid X receptor (RXR(gamma)) agonists using quantitative approach (CATALYST HypoRefine). One optimal pharmacophore model for selective RXR(gamma) agonists was determined through careful validation processes. The best quantitative model (Hypo-1) had five features and five excluded volumes: three hydrophobic aliphatic groups (HAL1, HAL2, and HAL3), one hydrophobic aromatic ring (HAR), and one hydrogen bond acceptor (HBA). The model was validated using a wide range of test molecules. It could predict agonist activity and identify highly potent molecules. The present results are valuable to discover and develop specific RXR(gamma) agonists with desired biological activities. PMID- 19121356 TI - Effects of beta-amyloid on behavioral and amino acids spectrum in rats' brain and their modulation by embryonic proteins. AB - One of the crucial events in the pathogenesis of neurodegenerative disorders linked with dementia-like Alzheimer's Disease (AD) is the disturbance in neurotransmission based on progressive deficit of neuromediators that is manifested by marked decrease in cognitive behavior, loss of memory and inability to learn as a result of impairment in synaptic plasticity of neurons. In this study we have used a new complex of proteoglycans of embryonic genesis (PEG) created by Prof. L. Mkrtchyan, as a possible therapeutic approach that can rescue neurons from further degeneration caused by beta-amyloid (Abeta). We attempt to reveal the biochemical (determination of neuroactive amino acids such as glutamate, GABA, taurine, glycine and aspartate) changes and behavior on Y-maze and avoidance/exploratory activity on elevated plus-maze task in rats' brain after modeling Alzheimer's disease by i.c.v. injection of Abeta25-35. Furthermore, in this study we analyzed the neuroprotective properties of PEG. Under the influence of PEG the concentration of all investigated amino acids both in cerebral cortex and hippocampus (except striatum changes) increased. In the present study we demonstrated that bilateral i.c.v. injection of aggregated Abeta25-35 in dosage 30nmol/rat resulted in impairment in spatial alternation behavior. Both preliminary (single) and double injection of PEG showed constant improvement of spatial memory after the first trial up to 90 days after i.c.v. injection of aggregated Abeta25-35. Our findings suggest that proteoglycans of embryonic genesis in neurodegenerative state show an expressed regulatory protective effect. PMID- 19121357 TI - Monitoring mouse serotonin transporter internalization in stem cell-derived serotonergic neurons by confocal laser scanning microscopy. AB - In the central nervous system serotonergic neurotransmission is terminated by the rapid removal of serotonin (5-hydroxytryptamine, 5HT) out of the extra-cellular space back into the presynaptic neuron. This task is fulfilled by a specific serotonin transporter (SERT) protein which controls the concentration of extra cellular 5HT. Consequently, one mechanism to regulate the efficacy of serotonergic neurotransmission is via modulation of the density of SERT molecules on the cell membrane. In this regard it has been shown, that chronic activation of the p38 mitogen-activated protein kinase (p38 MAPK) leads to enhanced SERT surface expression whereas activation of protein kinase C (PKC) reduces SERT surface expression. In addition, it has been reported that exposure to selective serotonin re-uptake inhibitors (SSRIs) leads to a down-regulation of SERT expression in vivo and in vitro in different cellular systems. Here, we have studied interactions between kinase- and SSRI-induced SERT internalization in mouse stem cell-derived serotonergic neurons expressing the native SERT allele in its natural surroundings. Therefore we established a method to quantify the amount of cell surface-expressed SERT molecules on individual cells by antibody detection combined with confocal laser scanning microscopy. Using this methodology we could show that activation of PKC, inhibition of the p38 MAPK as well as exposure to the SSRI citalopram each induced a significant reduction of cell surface-expressed SERT over time. Combinations of PKC activation, p38 MAPK inhibition and SSRI exposure led to a more pronounced down-regulation of SERT surface expression depending on the time of drug exposure. PMID- 19121359 TI - Hypothermia attenuates protective effects of ginkgolides on astrocytes from ischemia/reperfusion injury. AB - The neuroprotective roles of both hypothermia and ginkgolides have been well confirmed. We first examined whether hypothermia (32 or 28 degrees C) or ginkgolides have a protective effect on astrocytes against ischemia and reperfusion-induced injury. We demonstrated that ginkgolides, but not hypothermia, have a significantly time- and concentration-dependent protective role in ischemic astrocytes. We then investigated whether co-treatment with hypothermia and ginkgolides has a synergistic role to protect astrocytes against ischemia and reperfusion-induced injury. Cells were incubated with 18.75, 37.5 or 75 microg/ml of ginkgolides at 37, 32 or 28 degrees C for 24, 48 or 72 h before exposure to ischemia (24h) and then reperfusion (24h). Data showed that the co treatment induced a significant decrease, rather than an increase as we had expected, in their cellular viabilities and anti-apoptotic abilities as compared with the cells treated by ginkgolides only. Western blot analysis demonstrated that hypothermia (32 or 28 degrees C for 24h) has no effect on the expression of Hypoxia-inducible factor-1 alpha (HIF-1 alpha) protein, suggesting that HIF-1 alpha is not associated with the adverse effect of hypothermia on ginkgolides. The findings imply the importance of further investigating the effects of hypothermia on the pharmacological role or therapeutic efficacy of drugs commonly used clinically. PMID- 19121360 TI - Tardive dyskinesia associated with long-term administration of escitalopram and itopride in major depressive disorder. PMID- 19121358 TI - A genetic rat model of depression, Flinders sensitive line, has a lower density of 5-HT(1A) receptors, but a higher density of 5-HT(1B) receptors, compared to control rats. AB - Deficiencies in brain serotonergic neurotransmission, which is in part associated with the alteration of brain serotonin (5-HT) receptors, have been proposed as part of a neurochemical imbalance in affective disorders, including depression. The drugs used for the treatment of these disorders generally act through and/or on the serotonergic system. Different animal models of depression have provided researchers with tools to obtain a better understanding of drug actions and possibilities to obtain insight into the neurochemical bases of these disorders. The measurements of the 5-HT(1A) and 5-HT(1B) receptor densities in a rat model of depression, Flinders sensitive line (FSL) rats, and comparisons with Sprague Dawley (SPD) and Flinders resistant line (FRL) rats, are reported here. The receptor sites were quantified by autoradiography in more than 25 distinct brain regions known to have relatively large densities of respective sites. Some brain regions (e.g., dental gyrus, septal nucleus) were divided into several parts, according to previously known subdivisions, because of a substantial heterogeneity of these receptors. The densities in the FSL rats ("depressed" rats) were compared statistically to those in the SPD rats. In addition, comparisons were made to the densities in the FRL rats (rats not showing depressive symptoms). Comparisons were performed with the SPD and FRL rats because both of these strains have been used as control animals in studies of FSL rats. The results show that the densities of 5-HT(1A) receptors are not significantly different between the FSL and SPD rats, but they are significantly different from the FRL rats. 5-HT(1A) receptor density is significantly higher in the FRL rats than the SPD rats. The 5-HT(1B) receptors were significantly greater in the FSL rats than in either the SPD or FRL rats. In addition, the FRL rats have 5-HT(1B) receptor densities significantly lower in many brain regions than the SPD rats. The data presented here, in addition to previously reported differences in regional synthesis between these strains and the effect of acute citalopram on synthesis, suggest that SPD rats are likely a more appropriate control than FRL rats, when studies of FSL rats are performed with drugs acting directly or indirectly on, or through, the brain serotonergic system. However, comparisons, particularly of neurochemical and/or biological parameters in FRL rats, may reveal new insight into the alterations of 5-HT neurotransmission in this animal model of depression and possibly human depression, as well as the elevation of symptoms with treatments. The data also suggest that there could be a different fraction of 5-HT(1A) receptors in high and low affinity states in these strains, as well as the possibility of different intracellular signalling. PMID- 19121361 TI - Effects of acute and chronic administration of MK-801 on c-Fos protein expression in mice brain regions implicated in schizophrenia with or without clozapine. AB - This study investigated the effects of acute and chronic administration of the non-competitive NMDA receptor antagonists MK-801 on c-Fos protein expression in different brain regions of mice with or without clozapine. MK-801 (0.6 mg/kg) acute administration produced a significant increase in the expression of c-Fos protein in the layers III-IV of posterior cingulate and retrosplenial (PC/RS) cortex, which was consistent with the previous reports. Moreover, we presented a new finding that MK-801 (0.6 mg/kg) chronic administration for 8 days produced a significant increase of c-Fos protein expression in the PC/RS cortex, prefrontal cortex (PFC) and hypothalamus of mice. Among that, c-Fos protein expression in the PC/RS cortex of mice was most significant. Compared to acute administration, we found that MK-801 chronic administration significantly increased the expression of c-Fos protein in the PC/RS cortex, PFC and hypothalamus. Furthermore, pretreatment of mice with clozapine significantly decreased the expression of c-Fos protein induced by MK-801 acute and chronic administration. These results suggest that c-Fos protein, the marker of neuronal activation, might play an important role in the chronic pathophysiological process of schizophrenic model induced by NMDA receptor antagonist. PMID- 19121362 TI - Dietary exposure assessment of pregnant women to bisphenol-A from cans and microwave containers in Southern Spain. AB - Bisphenol-A (BPA), material for polycarbonate and epoxy resin synthesis, has been detected in canned food, among other food containers. In mammal studies, BPA transferred from mother to fetus, caused abnormality of reproductive organs, and advanced female puberty. BPA from canned food and microwave containers was analyzed using high performance liquid chromatography and gas chromatography-mass spectrometry. Population was cohort of mother-son pairs established at Granada University Hospital. Frequency of food consumption (including canned food) was studied with a semi-quantitative questionnaire. The most frequently consumed products were fish and juice cans, consumed 1-3 times/week by 34.7% and 22.3% of the study population, respectively. The women made little use of polymer microwave containers, 52.8% never using them and 45.9% using them <3 times/month. Estimated mean (standard deviation) intake of BPA was 1.1(0.839) microgram/day. No relationship was found between BPA exposure and maternal socio-demographic variables or newborn characteristics. This study offers the first estimate of BPA dietary intake by pregnant women in Southern Spain. The consumption of canned foods and drinks by these women means that their exposure was lower than EFSA estimates for the European population. Nevertheless it remains of concern, given the proven undesirable effects of low-level exposure and higher susceptibility of pregnant women. PMID- 19121363 TI - Characterization of the Trypanosoma cruzi ortholog of the SBDS protein reveals an intrinsically disordered extended C-terminal region showing RNA-interacting activity. AB - The human SBDS gene and its yeast ortholog SDO1 encode essential proteins that are involved in ribosome biosynthesis. SDO1 has been implicated in recycling of the ribosomal biogenesis factor Tif6p from pre-66S particles as well as in translation activation of 60S ribosomes. The SBDS protein is highly conserved, containing approximately 250 amino acid residues in animals, fungi and Archaea, while SBDS orthologs of plants and a group of protists contain an extended C terminal region. In this work, we describe the characterization of the Trypanosoma cruzi SBDS ortholog (TcSBDS). TcSBDS co-fractionates with polysomes in sucrose density gradients, which is consistent with a role in ribosome biosynthesis. We show that TcSBDS contains a C-terminal extension of 200 amino acids that displays the features of intrinsically disordered proteins as determined by proteolytic, circular dichroism and NMR analyses. Interestingly, the C-terminal extension is responsible for TcSBDS-RNA interaction activity in electrophoretic mobility shift assays. This finding suggests that Trypanosomatidae and possibly also other organisms containing SBDS with extended C-terminal regions have evolved an additional function for SBDS in ribosome biogenesis. PMID- 19121365 TI - Sleep to learn after stroke: implicit and explicit off-line motor learning. AB - After stroke, many individuals experience persistent motor impairments as well as altered patterns of sleep. Therefore, examining the role of sleep in motor skill learning following stroke is a critical issue. Other learning variables, such as type of instruction, may interact with sleep to influence sleep-dependent motor learning. Forty individuals with stroke and 40 control participants practiced a continuous motor tracking task and then either slept (sleep condition) or stayed awake (no-sleep condition) between practice and retention testing. Half were provided explicit information regarding the presence of a repeating sequence (explicit condition), while the other half were not (implicit condition). After stroke, individuals demonstrated sleep-dependent off-line motor learning of both the implicit and explicit version of the continuous tracking task; however, individuals with stroke who stayed awake between practice and retention testing did not demonstrate an improvement in motor performance at retention. Neither sleep nor instruction differentiated the performance of the healthy control participants. These data suggest that aspects of motor recovery after stroke may be modulated by sleep. PMID- 19121364 TI - A distributed neural system for top-down face processing. AB - Evidence suggests that the neural system associated with face processing is a distributed cortical network containing both bottom-up and top-down mechanisms. While bottom-up face processing has been the focus of many studies, the neural areas involved in the top-down face processing have not been extensively investigated due to difficulty in isolating top-down influences from the bottom up response engendered by presentation of a face. In the present study, we used a novel experimental method to induce illusory face-detection. This method allowed for directly examining the neural systems involved in top-down face processing while minimizing the influence of bottom-up perceptual input. A distributed cortical network of top-down face processing was identified by analyzing the functional connectivity patterns of the right fusiform face area (FFA). This distributed cortical network model for face processing includes both "core" and "extended" face processing areas. It also includes left anterior cingulate cortex (ACC), bilateral orbitofrontal cortex (OFC), left dorsolateral prefrontal cortex (DLPFC), left premotor cortex, and left inferior parietal cortex. These findings suggest that top-down face processing contains not only regions for analyzing the visual appearance of faces, but also those involved in processing low spatial frequency (LSF) information, decision-making, and working memory. PMID- 19121366 TI - Selective inhibition of cyclooxygenase-2 attenuates nitroglycerin-induced calmodulin-dependent protein kinase II alpha in rat trigeminal nucleus caudalis. AB - The nitric oxide donor, nitroglycerin (NTG) can trigger a migraine attack, after a delay of several hours in migraineurs, but not in healthy people. This long delay does not favor a pure vasodilatatory action. In rats, subcutaneous administration of NTG (10mg/kg) significantly and selectively increases the number of calmodulin-dependent protein kinase II alpha (CamKIIalpha) immunoreactive neurons in the trigeminal caudal nucleus (TNC) after 4h. The aim of our study was to determine if any isoforms of the cyclooxygenase (COX) enzyme might have a role in the NTG-induced increase of CamKIIalpha expression. In our experiments, we demonstrated that pretreatment with NS398, the selective COX-2 inhibitor attenuated the NTG-induced CamKIIalpha expression in the TNC at doses of 3 and 5mg/kg. In contrast, SC560, a selective COX-1 inhibitor failed to modulate this phenomenon in any of the dosages used (1, 5 and 10mg/kg). These findings suggest that COX-2, but not COX-1 derived metabolites are important factors in the NTG-induced CamKIIalpha expression. Thus this isoform may play a significant role in the induction of migraine. These data could help in the better understanding of the pathogenesis of headaches and the action of antimigraine drugs. PMID- 19121367 TI - Oculogyric crisis with exacerbation of psychosis: Possible mechanism and clinical implications. AB - Oculogyric crisis is a distressing acute/chronic side effect of neuroleptic medications. Chronic oculogyric crisis can be considered as a tardive hyperkinetic movement disorder and it may be associated with worsening of psychotic symptoms. Treatment strategies for chronic oculogyric crisis include; high potency antipsychotics and anticholinergics drugs for immediate relief and clozapine as a long-term treatment strategy. Here we are presenting case histories of four patients with oculogyric crisis and associated worsening of psychosis, its possible etiology and various treatment strategies. PMID- 19121368 TI - Enhanced adaptation of visual cortical cells to visual stimulation in aged cats. AB - Aging leads to compromised intracortical inhibition. Whether aging affects the adaptation of cortical neurons to sensory stimulation remains unknown. In this study, adaptation of V(1) neurons to visual stimuli was compared between young adult and aged cats using in vivo extracellular single-unit recording techniques. Our results indicated that neurons in aged cats exhibited significantly stronger adaptation to visual stimuli than did neurons in young adult cats. The enhanced adaptation of visual cortical cells to visual stimulation in the aging brain, which is of great significance to the energy savings in neuronal activity of senescent individuals, could be underlied by a simultaneous change of somatic afterhyperpolarization and synaptic depression mediated by an extensive age related GABAergic inhibition reduction at cortical and subcortical level. PMID- 19121369 TI - Transactivation by Runt related factor-2 of matrix metalloproteinase-13 in astrocytes. AB - We have previously shown functional expression by osteoblasts of signaling machineries required for neurotransmission in the brain. In this study, we have evaluated possible functional expression of different osseous genes in the brain. In embryonic and adult mouse brains, mRNA expression was invariably seen for the master regulator of osteoblastic differentiation Runt related factor-2 (Runx2), in addition to the partner protein core binding factor-beta and their targets such as osteopontin (OPN) and matrix metalloproteinase-13 (MMP13), but not for collagen-I or osteocalcin. In pluripotent P19 progenitor cells, Runx2 mRNA expression was drastically increased along with mRNA expression of an astrocytic marker, but not with neuronal marker mRNA expression. Both mRNA and corresponding protein were detected for Runx2 in cultured rat neocortical astrocytes and astrocytic C6 glioma cells. In C6 glioma cells, transient overexpression of Runx2 significantly increased mRNA expression of MMP13, but not of OPN. Moreover, transient overexpression of Runx2 significantly increased luciferase activity in C6 glioma cells transfected with the reporter plasmid linked to a wild-type Runx2 binding element in the MMP13 promoter, but not in cells with a mutated element. These results suggest that Runx2 signal input may lead to transactivation of MMP13 gene without affecting OPN expression in astrocytes. PMID- 19121370 TI - Oxidative stress-related aging: A role for prostate cancer? AB - Prostate cancer has the highest prevalence of any non-cutaneous cancer in the human body and essentially all men with circulating androgens will develop microscopic prostate cancer if they live long enough. Aging, considered as an impairment of body functions over time, caused by the accumulation of molecular damage in DNA, proteins and lipids, is also characterized by an increase in intracellular oxidative stress due to the progressive decrease of the intracellular ROS scavenging. The aging damage may eventually appear in age related health issues, which have a significant impact on the independence, general well-being and morbidity of the elderly. The association of aging with prostate cancer is undisputable as well as the association of aging with oxidative stress. Nevertheless, supportive evidence linking an increase in oxidative stress with prostate cancer is still scarce. This review is a comprehensive, literature-based analysis of the association of human prostate cancer with oxidative stress. The objective was to examine the involvement of reactive oxygen species in the mechanisms of prostatic carcinogenesis since the understanding of risk factors for prostate cancer has practical importance for public health, genetic and nutritional education, and chemoprevention. PMID- 19121373 TI - Directional information flows between brain hemispheres across waking, non-REM and REM sleep states: an EEG study. AB - The present electroencephalographic (EEG) study evaluated the hypothesis of a preferred directionality of communication flows between brain hemispheres across 24 h (i.e., during the whole daytime and nighttime), as an extension of a recent report showing changes in preferred directionality from pre-sleep wake to early sleep stages. Scalp EEGs were recorded in 10 normal volunteers during daytime wakefulness (eyes closed; first period: from 10:00 to 13:00 h; second period: from 14:00 to 18:00 h; third period: from 19:00 to 22:00 h) and nighttime sleep (four NREM-REM cycles). EEG rhythms of interest were delta (1-4 Hz), theta (5-7 Hz), alpha (8-11 Hz), sigma (12-15 Hz) and beta (16-28 Hz). The direction of the inter-hemispheric information flow was evaluated by computing the directed transfer function (DTF) from these EEG rhythms. Inter-hemispheric directional flows varied as a function of the state of consciousness (wake, NREM sleep, REM sleep) and in relation to different cerebral areas. During the daytime, alpha and beta rhythms conveyed inter-hemispheric signals with preferred Left-to-Right hemisphere direction in parietal and central areas, respectively. During the NREM sleep periods of nighttime, the direction of inter-hemispheric DTF information flows conveyed by central beta rhythms was again preponderant from Left-to-Right hemisphere in the stage 2, independent of cortical areas. No preferred direction emerged across the REM periods. These results support the hypothesis that specific directionality of communication flows between brain hemispheres is associated with wakefulness, NREM (particularly stage 2) and REM states during daytime and nighttime. They also reinforce the suggestive hypothesis of a relationship between inter-hemispheric directionality of EEG functional coupling and frequency of the EEG rhythms. PMID- 19121371 TI - Increased novelty-induced motor activity and reduced depression-like behavior in neuropeptide Y (NPY)-Y4 receptor knockout mice. AB - There is growing evidence that neuropeptide Y (NPY) acting through Y1 and Y2 receptors has a prominent role in modulating anxiety- and depression-like behavior in rodents. However, a role of other Y-receptors like that of Y4 receptors in this process is poorly understood. We now investigated male Y2, Y4 single and Y2/Y4 double knockout mice in behavioral paradigms for changes in motor activity, anxiety and depression-like behavior. Motor activity was increased in Y2, Y4 and Y2/Y4 knockout mice under changing and stressful conditions, but not altered in a familiar environment. Y4 and Y2 knockout mice revealed an anxiolytic phenotype in the light/dark test, marble burying test and in stress-induced hyperthermia, and reduced depression-like behavior in the forced swim and tail suspension tests. In Y2/Y4 double knockout mice, the response in the light/dark test and in the forced swim test was further enhanced compared with Y4 and Y2 knockout mice, respectively. High levels of Y4 binding sites were observed in brain stem nuclei including nucleus of solitary tract and area postrema. Lower levels were found in the medial amygdala and hypothalamus. Peripheral administration of pancreatic polypeptide (PP) induced Y4 receptor dependent c-Fos expression in brain stem, hypothalamus and amygdala. PP released peripherally from the pancreas in response to food intake, may act not only as a satiety signal but also modulate anxiety-related locomotion. PMID- 19121372 TI - Brain-derived neurotrophic factor modulates the severity of cognitive alterations induced by mutant huntingtin: involvement of phospholipaseCgamma activity and glutamate receptor expression. AB - The involvement of brain-derived neurotrophic factor (BDNF) in cognitive processes and the decrease in its expression in Huntington's disease suggest that this neurotrophin may play a role in learning impairment during the disease progression. We therefore analyzed the onset and severity of cognitive deficits in two different mouse models with the same mutant huntingtin but with different levels of BDNF (R6/1 and R6/1:BDNF+/- mice). We observed that BDNF modulates cognitive function in different learning tasks, even before the onset of motor symptoms. R6/1:BDNF+/- mice showed earlier and more accentuated cognitive impairment than R6/1 mice at 5 weeks of age in discrimination learning; at 5 weeks of age in procedural learning; and at 9 weeks of age in alternation learning. At the earliest age at which cognitive impairment was detected, electrophysiological analysis was performed in the hippocampus. All mutant genotypes showed reduced hippocampal long term potentiation (LTP) with respect to wild type but did not show differences between them. Thus, we evaluated the involvement of BDNF-trkB signaling and glutamate receptor expression in the hippocampus of these mice. We observed a decrease in phospholipaseCgamma activity, but not ERK, in R61, BDNF+/- and R6/1:BDNF+/- hippocampus at the age when LTP was altered. However, a specific decrease in the expression of glutamate receptors NR1, NR2A and GluR1 was detected only in R6/1:BDNF+/- hippocampus. Therefore, these results show that BDNF modulates the learning and memory alterations induced by mutant huntingtin. This interaction leads to intracellular changes, such as specific changes in glutamate receptors and in BDNF-trkB signaling through phospholipaseCgamma. PMID- 19121374 TI - Control of rodent and human spatial navigation by room and apparatus cues. AB - A growing body of literature indicates that rats prefer to navigate in the direction of a goal in the environment (directional responding) rather than to the precise location of the goal (place navigation). This paper provides a brief review of this literature with an emphasis on recent findings in the Morris water task. Four experiments designed to extend this work to humans in a computerized, virtual Morris water task are also described. Special emphasis is devoted to how directional responding and place navigation are influenced by room and apparatus cues, and how these cues control distinct components of navigation to a goal. Experiments 1 and 2 demonstrate that humans, like rats, perform directional responses when cues from the apparatus are present, while Experiment 3 demonstrates that place navigation predominates when apparatus cues are eliminated. In Experiment 4, an eyetracking system measured gaze location in the virtual environment dynamically as participants navigated from a start point to the goal. Participants primarily looked at room cues during the early segment of each trial, but primarily focused on the apparatus as the trial progressed, suggesting distinct, sequential stimulus functions. Implications for computational modeling of navigation in the Morris water task and related tasks are discussed. PMID- 19121376 TI - Nuclear gene targeting in Chlamydomonas as exemplified by disruption of the PHOT gene. AB - Chlamydomonas reinhardtii is the most powerful photosynthetic eukaryotic unicellular model organism. However, its potential is not fully exploitable since as in most green plants specific targeting of nuclear genes is not routinely possible. Recently, we have shown by repair of an introduced truncated model gene that transformation of Chlamydomonas with single stranded DNA greatly suppresses random integration of the DNA in the genome whereas homologous recombination (HR) is left unchanged. However, endogenous genes still could not be targeted. Here we present optimized transformation conditions that further improved HR and suppressed non-homologous DNA integration (NHI). The improved transformation strategy allowed us now to specifically inactivate in two different Chlamydomonas strains the nuclear PHOT gene, which encodes for the blue light photoreceptor phototropin (PHOT). The option to target moderately expressed Chlamydomonas nuclear genes with high efficiency now further improves the utility of this this alga for basic science and biotechnology. PMID- 19121375 TI - Soft-shell clam (Mya arenaria) p53: a structural and functional comparison to human p53. AB - The tumor suppressor p53 regulates genes involved in progression through the cell cycle, DNA repair, senescence or apoptosis in response to cell stress. Dysregulation of p53 can result in uncontrolled cellular proliferation. Invertebrate homologues to human p53 (Hsp53) have been identified, including a putative p53 gene (Map53) from the soft-shell clam (Mya arenaria). Predicted sequences for human and clam p53 proteins exhibit conservation in key domains. In light of this similarity, and the apparent dysregulation of Map53 under morphologically aberrant/pathologic conditions, we tested the hypothesis that the two proteins function in a similar manner. Plasmids expressing either Hsp53 or Map53 were introduced by transient transfection into the p53-null H1299 cell line. Functionality was assessed by monitoring the p53/mdm2 feedback loop and expression of p53-mediated downstream markers of growth arrest and apoptosis under non-stressed conditions. Hsp53 spontaneously induced markers of growth arrest, while Map53 expression induced neither cell arrest nor apoptosis. The difference in downstream activation is not likely the result of cytosolic sequestration since Map53, like Hsp53, localized almost exclusively to the nucleus. Functional similarity was observed in regulation by human MDM2, suggesting that the clam may have an mdm2 homologue. Protein modeling identified an apparent MDM2 binding site in Map53, supporting the observation of a potential Map53/MDM2 interaction. Significant amino acid differences present in the Map53 tetramerization domain may potentially affect p53 protein/protein interactions. Taken together, these data suggest that the Map53 shares some functional similarity with human p53 as well as with other invertebrates, positioning the mollusk at a critical juncture in evolution of this gene family. PMID- 19121377 TI - Conclusions and outlook. PMID- 19121378 TI - Toxicity studies of Tithonia diversifolia A. Gray (Asteraceae) in rats. AB - OBJECTIVE: To investigate the toxicity of an ethanolic extract of the aerial parts of Tithonia diversifolia, used in Nigeria to treat malaria, in rats. MATERIALS AND METHODS: A 70% ethanol extract was administered orally to adult Wistar rats at various dosages (400-1600 mg/kg) and the animals sacrificed and various organs examined at a range of times from 30 min up to 24 h after administration. RESULTS: The studies showed a dose- and time-dependent toxic effect, which was reversible on the kidney and liver while there was no noticeable adverse effect on the morphology of the heart, spleen and brain. CONCLUSION: A 70% ethanol extract of the aerial parts of Tithonia diversifolia, which had previously been shown to reduce parasitemia in mice infected with Plasmodium, displayed kidney and liver toxicity at the lowest dose tested. The use of this plant extract against malaria therefore raises concerns over its safety. PMID- 19121379 TI - The deficit in long-term potentiation induced by chronic administration of amyloid-beta is attenuated by treatment of rats with a novel phospholipid-based drug formulation, VP025. AB - Amyloid-beta (Abeta) peptides, the primary component of the amyloid plaques in Alzheimer's disease (AD), exert profound effects on neurons in vitro and negatively impact on neuronal function in vivo. One of the consequences of increased Abeta in the brain, either as a result of overexpression of the precursor amyloid precursor protein in transgenic mice, or injection into the brain is a decrease in one form of synaptic plasticity, long-term potentiation (LTP) in the hippocampus. Here we investigated the effect of infusion of Abeta for 28 days on LTP in dentate gyrus of rats and demonstrate that it was profoundly decreased compared with control-treated rats. We show that this effect is accompanied by increased activity of caspase 3, which is an indicator of cell stress. Significantly these changes were attenuated in animals which were pretreated with particles incorporating phosphatidylglycerol (VP025) and the evidence indicated that even when treatment was given 2 weeks after the start of the Abeta infusion, VP025 was capable of attenuating Abeta-induced changes. The evidence suggests that activation of caspase 3 was mediated by an Abeta-induced increase in sphingomyelinase, with the subsequent production of ceramide which is known to have a detrimental effect on neuronal function. PMID- 19121380 TI - Cerebral expression of DNA repair protein, Ku70, and its association with cell proliferation following cerebral hypoxia-ischemia in neonatal rats. AB - We hypothesized that increased Ku70 expression could be involved in recovery following cerebral hypoxia-ischemia. We investigated the progression of cerebral alterations in Ku70 expression at different time points (24 h, 72 h, 1 week, 4 weeks and 8 weeks) after hypoxia-ischemia (right carotid artery occlusion plus 1.5h of hypoxia) in neonatal rats. To determine whether in addition to its known role of DNA repair, Ku70 was associated with cell death or cell proliferation we performed double staining for Ku70 and DNA fragmentation or bromodeoxyuridine, respectively. The results show that Ku70 expression was increased in the infarct core and peri-infarct regions at 24h following hypoxia-ischemia. The increased Ku70 expression was transient in the infarct core with a loss of Ku70 positive cells over days. In contrast, in the peri-infarct region the expression of Ku70 remained increased at chronic times 8 weeks following the insult. Cells positive for DNA fragmentation were not co-localized with cells positive for Ku70 after an insult. However, most of the cells positive for bromodeoxyuridine indicative of cell proliferation were positive for Ku70 in the peri-infarct region at 8 weeks after the insult. Considering the roles of Ku70 in DNA repair or inhibiting apoptosis and its co-localization within cells that had undergone proliferation, Ku70 may be considered a potential novel target to enhance recovery following hypoxia-ischemia. PMID- 19121381 TI - Reconstruction of three-dimensional human skin model composed of dendritic cells, keratinocytes and fibroblasts utilizing a handy scaffold of collagen vitrigel membrane. AB - We previously we attempted to make a three-dimensional human skin model consisting of three different cells, dendritic cells, keratinocytes and fibroblasts (KDF-Skin) to evaluate immunoreactions in human skin; however, this model had various problems; for example (1) the incubation period for the construction of this model is long (about three weeks); (2) to construct the collagen gel, high amounts of fibroblasts are needed; and (3) the horny layer of keratinocytes in this skin model is thinner than that of keratinocytes in real human skin. In order to overcome these problems, a new three-dimensional human skin model utilizing a handy scaffold of collagen vitrigel membrane (VG-KDF-Skin) was constructed. As a result, after 14 days incubation, the epidermis layer of normal human keratinocytes was thicker than the keratinocyte layer of KDF-Skin. The incubation period for VG-KDF-Skin construction was 7 days shorter than that of KDF-Skin, and the number of fibroblasts needed to seed VG-KDF-Skin was four times fewer than that of KDF-Skin. After the application of sensitizers such as DNCB, VG-KDF-Skin induced the expression of CD86 and cytokine release. These results suggest that the new three-dimensional human skin model consisting of dendritic cells, keratinocytes, fibroblasts and collagen vitrigel membrane was more useful for alternative animal testing than the KDF-Skin model. PMID- 19121382 TI - Cytotoxic, genotoxic and oxidative stress induced by 1,4-naphthoquinone in B16F1 melanoma tumor cells. AB - Quinones have diverse pharmacological properties including antibacterial, antifungal, antiviral, anti-inflammatory, antipyretic and anticancer activity. The cytotoxic potential of 1,4-naphthoquinone (NQ14) was studied against B16F1 melanoma cells grown in vitro. NQ14 treatment resulted in a concentration dependent cytotoxicity as indicated by MTT assay and lactate dehydrogenase leakage assay. Depletion in cellular glutathione levels after 1h incubation with NQ14 correlated with the corresponding increase in reactive oxygen species generation as determined by 2',7'-dicholorofluorescein diacetate assay suggests the role of oxidative stress in cell death. The frequency of micronucleated binucleate cells increased with increasing doses of NQ14 with a corresponding decrease in the cytokinesis block proliferation index indicating the drug induced genotoxicity and cell division delay. Further, a dose-dependent decrease in the clonogenic cell survival indicated the potential of NQ14 to inhibit cell proliferation contributing to cell death. The cell death after NQ14 treatment may be attributed to apoptosis as seen in DNA ladder pattern along with necrosis as indicated in flow cytometric analysis of Annexin V/PI stained cells. Results of the present study demonstrate the cytotoxic and genotoxic potential of NQ14 by the induction of oxidative stress mediated mechanisms leading to tumor cell kill. PMID- 19121384 TI - Harmonization of terminology in developmental toxicology: the quest for a more precise description and a harmonized classification of fetal observations. AB - Harmonization of terminology in developmental toxicology is a prerequisite to ensure a better risk assessment of chemicals. As part of an international effort of the International Programme on Chemical Safety (IPCS) to harmonize terminology in developmental toxicology, workshops have taken place in Berlin since 1995. This publication reports the main outcomes of the Fifth and Sixth Berlin Workshops held in 2005 and 2007, respectively. The objective of the Fifth workshop was to discuss a draft international proposal for updating the glossary of descriptive terms for fetal abnormalities put forward by Wise et al. [Wise LD, et al. Terminology of developmental abnormalities in common laboratory mammals (version 1). Teratology 1997;55:249-92]. The participants were asked to classify the new external, visceral and skeletal observations included within this new version 2 of Terminology of Developmental Abnormalities in common Laboratory Mammals according to the two-category scheme (malformation and variation) agreed at previous Berlin workshops. The discussions held during the Sixth Workshop were mainly focused on the causes of uncertainty and low agreement regarding classification of some fetal observations as malformations or variations. Lack of precision in descriptive terms and insufficient knowledge of the postnatal consequences of fetal observations had been identified as major causes of uncertainty and lower agreement among evaluators regarding the classification of "grey zone anomalies", i.e. abnormalities that do not fit readily into one of the two categories (malformation or variation). Imprecise anatomical terms, observation terms that are too broad, lack of information on severity and the use of different terms for the same change or different severities of the same change, were found to be the main reasons that descriptive terms are often not sufficiently precise to allow accurate classification of findings. It was agreed that provision of additional information, including sub-location within the affected structure, more detailed description of the nature of the change, in conjunction with presentation of photographs wherever possible, and a grading for severity would make descriptive terms more precise, thereby reducing misclassifications. A better knowledge of the adversity and postnatal consequences of fetal observations was considered as the key issue for achieving a substantial reduction in the number of misclassifications and grey zone anomalies. The urgent need for additional research along this line as a prerequisite for a better risk assessment was emphasized by the participants. PMID- 19121383 TI - Discovery of transcriptional regulators and signaling pathways in the developing pituitary gland by bioinformatic and genomic approaches. AB - We report a catalog of the mouse embryonic pituitary gland transcriptome consisting of five cDNA libraries including wild type tissue from E12.5 and E14.5, Prop1(df/df) mutant at E14.5, and two cDNA subtractions: E14.5 WT-E14.5 Prop1(df/df) and E14.5 WT-E12.5 WT. DNA sequence information is assembled into a searchable database with gene ontology terms representing 12,009 expressed genes. We validated coverage of the libraries by detecting most known homeobox gene transcription factor cDNAs. A total of 45 homeobox genes were detected as part of the pituitary transcriptome, representing most expected ones, which validated library coverage, and many novel ones, underscoring the utility of this resource as a discovery tool. We took a similar approach for signaling-pathway members with novel pituitary expression and found 157 genes related to the BMP, FGF, WNT, SHH and NOTCH pathways. These genes are exciting candidates for regulators of pituitary development and function. PMID- 19121385 TI - Curcumin protects retinal cells from light-and oxidant stress-induced cell death. AB - Age-related macular degeneration (AMD) is a complex disease that has potential involvement of inflammatory and oxidative stress-related pathways in its pathogenesis. In search of effective therapeutic agents, we tested curcumin, a naturally occurring compound with known anti-inflammatory and antioxidative properties, in a rat model of light-induced retinal degeneration (LIRD) and in retina-derived cell lines. We hypothesized that any compound effective against LIRD, which involves significant oxidative stress and inflammation, would be a candidate for further characterization for its potential application in AMD. We observed significant retinal neuroprotection in rats fed diets supplemented with curcumin (0.2% in diet) for 2 weeks. The mechanism of retinal protection from LIRD by curcumin involves inhibition of NF-kappaB activation and down-regulation of cellular inflammatory genes. When tested on retina-derived cell lines (661W and ARPE-19), pretreatment of curcumin protected these cells from H(2)O(2) induced cell death by up-regulating cellular protective enzymes, such as HO-1, thioredoxin. Since, curcumin with its pleiotropic activities can modulate the expression and activation of many cellular regulatory proteins such as NF-kappaB, AKT, NRF2, and growth factors, which in turn inhibit cellular inflammatory responses and protect cells; we speculate that curcumin would be an effective nutraceutical compound for preventive and augmentative therapy of AMD. PMID- 19121386 TI - Identification of paraoxonase-1 in induced sputum from healthy volunteers. PMID- 19121387 TI - Distinct intracellular signaling pathways control the synthesis of IL-8 and RANTES in TLR1/TLR2, TLR3 or NOD1 activated human airway epithelial cells. AB - Inflammation is a central feature of many respiratory diseases. Airway epithelial cells are exposed to many agents present in the air that can alter their function and have important structural consequences for the airways. In this study, 19 Toll-Like Receptors (TLRs) and Nucleotide-binding Oligomerization Domain (NOD)1/NOD2 ligands were screened for their capacity to up-regulate Interleukin-8 (IL-8) and Regulated upon Activation Normal T cell Expressed and Secreted (RANTES) in airway epithelial cells. Three ligands (Pam3CSK4, Poly I:C and C12-ie DAP) were selected for their capacity to activate different receptor complexes (TLR1/TLR2, TLR3 and NOD1 respectively) while leading to the increase of both IL 8 and RANTES albeit with distinct kinetics. Using protein kinase inhibitors we found that the Nuclear Factor kappaB (NFkappaB) pathway is essential for the transcriptional regulation of both IL-8 and RANTES following the activation of TLR1/TLR2, TLR3 and NOD1. In contrast, the Mitogen-Activated Protein Kinases (MAPKs) Extracellular signal-Regulated Kinase (ERK)1/ERK2 and p38 MAPK were necessary for the transcription of IL-8 but not RANTES. Moreover, we found that the p38 MAPK was implicated in the post-transcriptional regulation of IL-8 following TLR3 activation. The distinction made between pathways involved in the regulation of IL-8 and RANTES gives rise to the possibility of designing more targeted clinical approaches based on the biological functions to be ablated. PMID- 19121388 TI - Feasibility, stability and release performance of a time-dependent insulin delivery system intended for oral colon release. AB - The aim of the present work was to evaluate the viability of a time-dependent delivery platform (Chronotopic) in preparing an insulin-based system intended for oral colon delivery. The main objectives were to assess the influence of the manufacturing process and storage conditions on the protein stability. Insulin loaded cores were manufactured by direct compression and were subsequently coated with hydroxypropyl methylcellulose (HPMC) in a top-spray fluid bed up to increasing weight gains, namely 20%, 60% and 100%. In order to evaluate the impact the operating conditions may have on the protein integrity, insulin and its main degradation products (A21-desamido insulin -A21, Other Insulin-Related Compounds -OIRCs, and High-Molecular Weight Proteins -HMWPs) were assayed on samples collected after each process step by chromatographic methods. Furthermore, long-term (4 degrees C) and accelerated (25 degrees C-60% RH) stability studies were carried out on tablet cores and coated systems by assessing insulin, A21, OIRC and HMWP percentages throughout a one-year storage period. In addition, the in vitro release behaviour was investigated during the same study period. The overall results indicated that the manufacturing process is not detrimental for insulin integrity and that 4 degrees C storage temperature alters neither the protein content nor the release performances of the device. It was therefore concluded that insulin-containing systems intended for oral colon delivery can be obtained by the Chronotopic technology. PMID- 19121389 TI - Mission to steroids. PMID- 19121391 TI - Enhanced expression of ERalpha in astrocytes modifies the response of cortical neurons to beta-amyloid toxicity. AB - Estrogen receptor alpha (ERalpha) is over-expressed in reactive glia under conditions of neuronal damage. To elucidate the functional significance of ERalpha overexpression, an in vitro model of reactive astrocytes with enhanced expression of ERalpha was obtained by growth in G5 culture supplement. Exposure of cortical neurons to beta-amyloid in the presence of either conditioned medium from reactive astrocytes previously treated with 17beta-estradiol (17betaE2) or transferring of 17betaE2-pretreated astrocytes, caused a greater neuroprotective effect compared to the respective control conditions, although reactive glia resulted being per se neuroprotective. Blockade of ERalpha overexpression by the ER antagonist ICI182,780 was not successful as ICI182,780 behaved as an agonist. However, complete prevention of 17betaE2 effect by ICI182,780 produced an increased sensitivity of neurons to beta-amyloid toxicity. A similar effect was observed when ERalpha knock-down was induced by siRNA. It is suggested that increased ERalpha expression in reactive glia may have a role in limiting neuronal damage. PMID- 19121390 TI - The genome of a lepidopteran model insect, the silkworm Bombyx mori. AB - Bombyx mori, the domesticated silkworm, is a major insect model for research, and the first lepidopteran for which draft genome sequences became available in 2004. Two independent data sets from whole-genome shotgun sequencing were merged and assembled together with newly obtained fosmid- and BAC-end sequences. The remarkably improved new assembly is presented here. The 8.5-fold sequence coverage of an estimated 432 Mb genome was assembled into scaffolds with an N50 size of approximately 3.7 Mb; the largest scaffold was 14.5 million base pairs. With help of a high-density SNP linkage map, we anchored 87% of the scaffold sequences to all 28 chromosomes. A particular feature was the high repetitive sequence content estimated to be 43.6% and that consisted mainly of transposable elements. We predicted 14,623 gene models based on a GLEAN-based algorithm, a more accurate prediction than the previous gene models for this species. Over three thousand silkworm genes have no homologs in other insect or vertebrate genomes. Some insights into gene evolution and into characteristic biological processes are presented here and in other papers in this issue. The massive silk production correlates with the existence of specific tRNA clusters, and of several sericin genes assembled in a cluster. The silkworm's adaptation to feeding on mulberry leaves, which contain toxic alkaloids, is likely linked to the presence of new-type sucrase genes, apparently acquired from bacteria. The silkworm genome also revealed the cascade of genes involved in the juvenile hormone biosynthesis pathway, and a large number of cuticular protein genes. PMID- 19121392 TI - Regulation of contractile activity by magnolol in the rat isolated gastrointestinal tracts. AB - This study examined the pharmacological property of magnolol, a phenolic compound purified from Magnolia officinalis, on the GI motility using the rat isolated gastrointestinal (GI) strips. Magnolol (0.3-30 microM) dose-dependently stimulated the tone and amplitude of spontaneous contractions in ileum longitudinal muscles. Magnolol at 3 microM significantly increased the contractions of jejunum longitudinal and colon circular muscles, but not the longitudinal muscle contractions in fundus, antrum and colon. Pretreatment of ileum strips with either atropine (0.5 microM) or 4-diphenyllacetoxy-N(2 chloriethyl)-piperidine (4-DAMP, 0.5 microM) dramatically inhibited the acetylcholine (ACh, 0.1 microM)- and magnolol (3 microM)-induced longitudinal muscle contractions, but they were not affected by methoctramine (0.5 microM) and hexamethonium (0.5 microM). Ondansetron (0.1 microM) and GR113808 (2 microM) significantly reduced the tone of ileum longitudinal muscle contractions stimulated by 5-HT (10 microM), but not the amplitude. Magnolol (3 microM) induced ileum longitudinal muscle contractions, both tone and amplitude, were significantly blocked by GR113808, but not by ondansetron. Taken together, magnolol differently regulates the spontaneous GI motility according to the region of GI tracts and orientation of smooth muscles, and magnolol-induced regulation of smooth muscle contractions in rat GI strips is likely to be mediated, at least in part, by activation of ACh and 5-HT receptors, possibly the M(3) and/or 5-HT(4) receptors. PMID- 19121393 TI - Leptin inhibits glycogen catabolism but does not modify acutely the suppressive effect of insulin on glucose production and glycogenolysis stimulated by 8-Br cAMP in rat liver perfused in situ. AB - Leptin, a hormone secreted by the adipocytes, plays a central role in glucose metabolism and the action of insulin. Here we assessed, by means of rat-liver perfusion, the direct influence of physiological (10 ng/ml) and supraphysiological (50 or 100 ng/ml) concentrations of leptin on the suppressive effect of insulin on the glucose production and glycogenolysis stimulated by 8 bromoadenosine-3':5'-monophosphate (8-Br-cAMP). Portal infusion of insulin (20 microU/ml) or leptin (10 ng/ml) reduced (p<0.05) the glucose production and glycogenolysis induced by 8-Br-cAMP (0.3 microM). However, portal infusion of physiological (10 ng/ml) and supraphysiological (50 or 100 ng/ml) concentrations of leptin together with the insulin did not modify the suppressive effect of the latter on the glucose production and glycogenolysis stimulated by 8-Br-cAMP. Moreover, prolonging the period of leptin infusion from 20 to 40 min also failed to influence the liver response to insulin. Thus, we conclude that: (a) leptin, at physiological levels, has a direct and acute effect, inhibiting the glucose production and glycogenolysis stimulated by 8-Br-cAMP; (b) leptin, at either physiological or supraphysiological concentrations, has no short-term influence on the suppressive effect of insulin on glycogen catabolism stimulated by 8-Br cAMP. PMID- 19121394 TI - Chitosan oligosaccharides attenuate hydrogen peroxide-induced stress injury in human umbilical vein endothelial cells. AB - Chitosan oligosaccharides (COS) have been reported to have anticancer activity, immuno-enhancing effect and antimicrobial activity. However, other biological activities are unknown. Herein, we investigated the protective effects of COS against hydrogen peroxide (H(2)O(2))-induced oxidative damage on human umbilical vein endothelial cells (HUVEC, ECV304 cells). After 24h pre-incubation with COS (25-200 microg/ml), the viability loss in ECV304 cells induced by H(2)O(2) (300 microM) for 12h was markedly restored in a concentration-dependent manner as measured by MTT assay. This effect was accompanied by a marked decrease in intracellular reactive oxygen species (ROS) by measuring intensity of DCFH fluorescence. COS also exerted preventive effects on suppressing the production of lipid peroxidation such as malondialdehyde (MDA), restoring activities of endogenous antioxidants including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), along with the capacity of increasing levels of nitric oxide (NO) and nitric oxide synthase (NOS), as were determined by commercial regent kits. In addition, pre-incubation of COS with ECV304 cells for 24h resulted in the reduction of apoptosis and the induction of cell cycle arrest in G(1)/S+M phase as assayed quantitatively by Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit using flow cytometry. Taken together, our findings suggest that COS can effectively protect HUVECs against oxidative stress by H(2)O(2), which might be of importance in the treatment of cardiovascular diseases. PMID- 19121395 TI - Cloning and expression of carp acetylcholinesterase gene in Pichia pastoris and characterization of the recombinant enzyme. AB - The gene encoding acetylcholinesterase (AChE) was cloned from common carp muscle tissue. The full-length cDNA was 2368 bp that contains a coding region of 1902 bp, corresponding to a protein of 634 amino acids. The deduced amino acid sequence showed a significant homology with those of ichthyic AChEs and several common features among them, including T peptide encoded by exon T in the C terminus. Three yeast expression vectors were constructed and introduced into the yeast Pichia pastoris. The transformant harboring carp AChE gene lacking exon T most effectively produced AChE activity extracellularly. The replacement of the native signal sequence with the yeast alpha-factor prepro signal sequence rather decreased the production. A decrease in cultivation temperature from 30 to 15 degrees C increased the activity production 32.8-fold. The purified recombinant AChE lacking T peptide, eluted as a single peak with a molecular mass of about 230 kDa on the gel filtration chromatography, exhibited the specific activity of 4970 U/mg. On the SDS-PAGE, three proteins with molecular masses of 73, 54, and 22 kDa were observed. These proteins were N-glycosylated, and their N-terminal sequence showed that the latter two were produced from the former probably by proteolytic cleavage at the C-terminal region. Thus, the recombinant AChE is homotrimer of three identical subunits with 73 kDa. The optimal temperature and pH of the recombinant were comparable to those of the native enzyme purified previously, but the values of kinetic parameters and the sensitivities to substrate inhibition and inhibitors were considerably different between them. PMID- 19121396 TI - Expression and purification of human TRPV1 in baculovirus-infected insect cells for structural studies. AB - TRPV1 is a ligand-gated cation channel that is involved in acute thermal nociception and neurogenic inflammation. By using the GP67 signal peptide, high levels of full-length human TRPV1 was expressed in High Five insect cells using the baculovirus expression system. The functional activity of the expressed TRPV1 was confirmed by whole-cell ligand-gated ion flux recordings in the presence of capsaicin and low pH and via specific ligand binding to the isolated cellular membranes. Efficient solubilization and purification protocols have resulted in milligram amounts of detergent-solubilized channel at 80-90% purity after Ni2+ IMAC chromatography and size exclusion chromatography. Western blot analysis of amino and carboxyl terminal domains and MS of tryptic digestions of purified protein confirmed the presence of the full-length human TRPV1. Specific ligand binding experiments confirmed the protein integrity of the purified human TRPV1. PMID- 19121397 TI - Expression and purification of NEIL3, a human DNA glycosylase homolog. AB - The base excision repair (BER) pathway is mainly responsible for the repair of a vast number of non-bulky lesions produced by alkylation, oxidation or deamination of bases. DNA glycosylases are the key enzymes that recognize damaged bases and initiate BER by catalyzing the cleavage of the N-glycosylic bond between the base and the sugar. Many of the mammalian DNA glycosylases have been identified by a combination of biochemical and bioinformatics analysis. Thus, a mammalian family of three proteins (NEIL1, NEIL2 and NEIL3) that showed homology to the Escherichia coli Fpg/Nei DNA glycosylases was identified. Two of the proteins, NEIL1 and NEIL2 have been thoroughly characterized and shown to initiate BER of a diverse number of oxidized lesions. However, much less is known about NEIL3. The biochemical properties of NEIL3 have not been elucidated. This is mainly due to the difficulty in the expression and purification of NEIL3. Here, we describe the expression and partial purification of full-length human NEIL3 and the expression, purification and characterization of a truncated human core-NEIL3 (amino acids 1-301) that contains the complete E. coli Fpg/Nei-like domain but lacks the C-terminal region. PMID- 19121399 TI - Dynamics of sheet nacre formation in bivalves. AB - Formation of nacre (mother-of-pearl) is a biomineralization process of fundamental scientific as well as industrial importance. However, the dynamics of the formation process is still not understood. Here, we use scanning electron microscopy and high spatial resolution ion microprobe depth-profiling to image the full three-dimensional distribution of organic materials around individual tablets in the top-most layer of forming nacre in bivalves. Nacre formation proceeds by lateral, symmetric growth of individual tablets mediated by a growth ring rich in organics, in which aragonite crystallizes from amorphous precursors. The pivotal role in nacre formation played by the growth-ring structure documented in this study adds further complexity to a highly dynamical biomineralization process. PMID- 19121398 TI - Heparin binding induces conformational changes in Adeno-associated virus serotype 2. AB - Adeno-associated virus serotype 2 (AAV2) uses heparan sulfate proteoglycan as a cell surface-attachment receptor. In this study the structures of AAV2 alone and complexed with heparin were determined to approximately 18A resolution using cryo electron microscopy and three-dimensional image reconstruction. A difference map showed positive density, modeled as heparin, close to the icosahedral twofold axes and between the protrusions that surround the threefold axes of the capsid. Regions of the model near the threefold place the receptor in close proximity to basic residues previously identified as part of the heparin binding site. The region of the model near the twofold axes identifies a second contact site, not previously characterized but which is also possibly configured by heparin binding. The difference map also revealed two significant conformational changes: (I) at the tops of the threefold protrusions, which have become flattened in the complex, and (II) at the fivefold axes where the top of the channel is widened possibly in response to movement of the HI loops in the capsid proteins. Ordered density in the interior of the capsid in the AAV2-heparin complex was interpreted as nucleic acid, consistent with the presence of non-viral DNA in the expressed capsids. PMID- 19121400 TI - In vitro and in vivo immune responses induced by a DNA vaccine encoding the VP2 gene of the infectious pancreatic necrosis virus. AB - The work presented here describes the construction of a plasmid encoding the VP2 gene of the infectious pancreatic necrosis virus (IPNV), its expression in BF-2 cells and an evaluation of its activity in brown trout (Salmo trutta L) soon after injection. Preliminary experiments to evaluate the potential of the plasmid to induce neutralizing antibodies were also performed. We established a BF-2 cell line that expresses VP2 constitutively and we examined the infection of these VP2 transfected BF-2 cells with homologous and heterologous viruses. The expression kinetics of IFN, and of the IFN-induced genes Mx and ISG15, was also evaluated in brown trout over a 15 day interval, and quantified by real-time or semi quantitative PCR. Type I IFN and Mx are markers of the non-specific innate immune response to viruses and they are involved in antiviral defences. Our results demonstrate that expression of the IPNV VP2 protein in BF-2 cells induces an antiviral state against IPNV and against the infectious haematopoietic necrosis virus (IHNV). In BF-2 infected cells, VP2 inhibited both the IPNV and IHNV induced cytopathic effect to some extent, as well as the virus yield. In vivo, VP2 was expressed in haematopoietic tissues such as the head kidney of 7 month old trout. In addition, it induced early immune responses and specific immunity 30 days after injection. IFN mRNA expression increased sharply on the 1st and 15th day post-injection and expression of other IFN-induced genes as Mx and ISG15 was also detected soon after vaccination of brown trout. Moreover, specific antibodies were detected 30 days after vaccination. These results suggest that the VP2 gene is a good candidate for the design of IPNV-DNA vaccines and to investigate the use of cytokines as co-stimulatory molecules. PMID- 19121401 TI - Modeling brain activation patterns for the default and cognitive states. AB - We argue that spatial patterns of cortical activation observed with EEG, MEG and fMRI might arise from spontaneous self-organisation of interacting populations of excitatory and inhibitory neurons. We examine the dynamical behavior of a mean field cortical model that includes chemical and electrical (gap-junction) synapses, focusing on two limiting cases: the "slow-soma" limit with slow voltage feedback from soma to dendrite, and the "fast-soma" limit in which the feedback action of soma voltage onto dendrite reversal potentials is instantaneous. For slow soma-dendrite feedback, we find a low-frequency (approximately 1 Hz) dynamic Hopf instability, and a stationary Turing instability that catalyzes formation of patterned distributions of cortical firing-rate activity with pattern wavelength approximately 2 cm. Turing instability can only be triggered when gap-junction diffusion between inhibitory neurons is strong, but patterning is destroyed if the tonic level of subcortical excitation is raised sufficiently. Interaction between the Hopf and Turing instabilities may describe the non-cognitive background or "default" state of the brain, as observed by BOLD imaging. In the fast-soma limit, the model predicts a high-frequency Hopf (approximately 35 Hz) instability, and a traveling-wave gamma-band instability that manifests as a 2-D standing-wave pattern oscillating in place at approximately 30 Hz. Small levels of inhibitory diffusion enhance and broaden the definition of the gamma antinodal regions by suppressing higher-frequency spatial modes, but gamma emergence is not contingent on the presence of inhibitory gap junctions; higher levels of diffusion suppress gamma activity. Fast-soma instabilities are enhanced by increased subcortical stimulation. Prompt soma-dendrite feedback may be an essential component of the genesis and large-scale cortical synchrony of gamma activity observed at the point of cognition. PMID- 19121402 TI - Reconsideration of systematic relationships within the order Euplotida (Protista, Ciliophora) using new sequences of the gene coding for small-subunit rRNA and testing the use of combined data sets to construct phylogenies of the Diophrys complex. AB - Comprehensive molecular analyses of phylogenetic relationships within euplotid ciliates are relatively rare, and the relationships among some families remain questionable. We performed phylogenetic analyses of the order Euplotida based on new sequences of the gene coding for small-subunit RNA (SSrRNA) from a variety of taxa across the entire order as well as sequences from some of these taxa of other genes (ITS1-5.8S-ITS2 region and histone H4) that have not been included in previous analyses. Phylogenetic trees based on SSrRNA gene sequences constructed with four different methods had a consistent branching pattern that included the following features: (1) the "typical" euplotids comprised a paraphyletic assemblage composed of two divergent clades (family Uronychiidae and families Euplotidae-Certesiidae-Aspidiscidae-Gastrocirrhidae), (2) in the family Uronychiidae, the genera Uronychia and Paradiophrys formed a clearly outlined, well-supported clade that seemed to be rather divergent from Diophrys and Diophryopsis, suggesting that the Diophrys-complex may have had a longer and more separate evolutionary history than previously supposed, (3) inclusion of 12 new SSrRNA sequences in analyses of Euplotidae revealed two new clades of species within the family and cast additional doubt on the present classification of genera within the family, and (4) the intraspecific divergence among five species of Aspidisca was far greater than those of closely related genera. The ITS1-5.8S ITS2 coding regions and partial histone H4 genes of six morphospecies in the Diophrys-complex were sequenced along with their SSrRNA genes and used to compare phylogenies constructed from single data sets to those constructed from combined sets. Results indicated that combined analyses could be used to construct more reliable, less ambiguous phylogenies of complex groups like the order Euplotida, because they provide a greater amount and diversity of information. PMID- 19121404 TI - Sex determination in flatfishes: Mechanisms and environmental influences. AB - Flounder of the genus Paralichthys exhibit a unique mode of sex determination where both low and high temperatures induce male-skewed sex ratios, while intermediate temperatures produce a 1:1 sex ratio. Male differentiation is thus easily induced in genetic females creating a combination of genetic (GSD) and environmental sex determination (ESD). Since male flounder become reproductively fit at substantially smaller body sizes than females, temperature or other environmental variables that elicit lower growth rates may also influence sex differentiation toward male development. This review covers our current knowledge of sex determination and differentiation in flatfishes including possible adaptive significance of ESD and involvement of factors such as aromatase (cyp19). PMID- 19121403 TI - Flow cytometric analysis of BrdU incorporation as a high-throughput method for measuring adult neurogenesis in the mouse. AB - INTRODUCTION: The generation of new neurons occurs throughout adulthood in discrete brain regions, and may be regulated by neuropsychiatric diseases and therapeutic drug treatments. Most current methods that study this process measure the labeling of newborn cells by 5-bromo-2-deoxyuridine (BrdU) using immunohistochemical methods followed by the microscopic counting of BrdU positive cells. This method is time consuming and labor intensive, typically taking several weeks to analyze. METHODS: Therefore, we characterized a method to measure BrdU incorporation in the adult mouse hippocampus in vivo by using flow cytometry, which normally allows analysis of data within a single day. RESULTS: The present study compared multiple BrdU dosing and loading protocols to determine a dosing strategy that produced the best signal to noise ratio. BrdU incorporation was also compared across different brain regions. The method was sensitive to a number of experimental disease manipulations. Induction of type-1 diabetes and depletion of norepinephrine reduced hippocampal cell proliferation. In contrast, chronic administration of electroconvulsive shock, a somatic treatment for depression, as well as chronic treatment with the antidepressant fluoxetine elevated hippocampal cell proliferation. This increase in cell proliferation with fluoxetine was detected as early as 14 days into treatment. Moreover, comparing measures of cell proliferation obtained by immunohistochemical and flow cytometric methods within the same animals were convergent and significantly correlated to each other. Flow cytometry was also sufficiently sensitive to quantify the survival of newly born cells. DISCUSSION: These experiments validate the utility of flow cytometry in analyzing hippocampal cell proliferation and survival in a reliable and high-throughput fashion. The speedy analysis afforded by flow cytometry lends itself to be utilized in novel drug discovery and physiology. PMID- 19121405 TI - Eosinophilic airway disorders associated with chronic cough. AB - Chronic cough is a major clinical problem. The causes of chronic cough can be categorized into eosinophilic and noneosinophilic disorders, the former being comprised of asthma, cough variant asthma (CVA), atopic cough (AC) and non asthmatic eosinophilic bronchitis (NAEB). Cough is one of the major symptoms of asthma. Cough in asthma can be classified into three categories; 1) CVA: asthma presenting solely with coughing, 2) cough-predominant asthma: asthma predominantly presenting with coughing but also with dyspnea and/or wheezing, and 3) cough remaining after treatment with inhaled corticosteroid (ICS) and beta2 agonists in patients with classical asthma, despite control of other symptoms. There may be two subtypes in the last category; one is cough responsive to anti mediator drugs such as leukotriene receptor antagonists and histamine H1 receptor antagonists, and the other is cough due to co-morbid conditions such as gastroesophageal reflux. CVA is one of the commonest causes of chronic isolated cough. It shares a number of pathophysiological features with classical asthma with wheezing such as atopy, airway hyperresponsiveness (AHR), eosinophilic airway inflammation and various features of airway remodeling. One third of adult patients may develop wheezing and progress to classical asthma. As established in classical asthma, ICS is considered the first-line treatment, which improves cough and may also reduce the risk of progression to classical asthma. AC proposed by Fujimura et al. presents with bronchodilator-resistant dry cough associated with an atopic constitution. It involves eosinophilic tracheobronchitis and cough hypersensitivity and responds to ICS treatment, while lacking in AHR and variable airflow obstruction. These features are shared by non asthmatic eosinophilic bronchitis (NAEB). However, atopic cough does not involve bronchoalveolar eosinophilia, has no evidence of airway remodeling, and rarely progresses to classical asthma, unlike CVA and NAEB. Histamine H1 antagonists are effective in atopic cough, but their efficacy in NAEB is unknown. AHR of NAEB may improve with ICS within the normal range. Taken together, NAEB significantly overlaps with atopic cough, but might also include milder cases of CVA with very modest AHR. The similarity and difference of these related entities presenting with chronic cough and characterized by airway eosinophilia will be discussed. PMID- 19121406 TI - IL-17-producing T lymphocytes in lung tissue and in the bronchoalveolar space after exposure to endotoxin from Escherichia coli in vivo--effects of anti inflammatory pharmacotherapy. AB - Interleukin (IL)-17 may play a critical role for the innate immune response in mammals. However, little is known about its production in T lymphocytes in comparison with other cells, in lung tissue and in the bronchoalveolar space in vivo. Even less is known about the effects of anti-inflammatory pharmacotherapy on this IL-17 production. In this study on mice we show that one single, intranasal exposure to endotoxin from Escherichia coli increases extracellular IL 17 protein in bronchoalveolar (BAL) samples during 3 days, and is accompanied by a local increase in neutrophils and other inflammatory cells. This endotoxin exposure also elevates IL-17 mRNA in lung tissue samples. Moreover, after endotoxin exposure, the absolute number of CD3-positive cells containing intracellular IL-17 protein is increased as well; from a moderate cell number in lung tissue samples and from virtually none in BAL samples; with the number in lung tissue exceeding that observed in BAL samples. Notably, we also demonstrate that among the cells that contain intracellular IL-17 protein after endotoxin exposure, the percentage of CD3-positive cells is similar to that of CD3-negative cells in lung tissue. In contrast, CD3-negative cells dominate among IL-17 containing cells in BAL samples. A high systemic dose of a glucocorticoid receptor agonist attenuates the endotoxin-induced increase in extracellular IL-17 protein in BAL samples, IL-17 mRNA in lung tissue samples, and in IL-17 containing CD3-positive cells in BAL and lung tissue samples. This is also true for the endotoxin-induced accumulation of neutrophils and other inflammatory BAL cells in vivo. A systemic dose of a calcineurin phosphatase inhibitor exerts a less complete and more selective effect on the endotoxin-induced increase in extracellular IL-17 protein and on neutrophils in BAL samples. In vitro, endotoxin also increases extracellular IL-17 protein in a co-culture of CD3 positive spleen cells and adherent mononuclear BAL cells; an increase that was inhibited by a glucocorticoid as well as by a calcineurin phosphatase inhibitor. In conclusion, endotoxin-induced IL-17 production and release from T lymphocytes originates from cells that reside in lung tissue and from cells that have been recruited to the bronchoalveolar space. In both compartments, there is also a substantial number of cells other than T lymphocytes that contain IL-17 after endotoxin exposure. The sustained IL-17 production from T lymphocytes and the associated neutrophil accumulation may be inhibited non-selectively through glucocorticoid receptor stimulation and more selectively through calcineurin phosphatase inhibition. PMID- 19121408 TI - The genetics of a pharma merger. AB - The 1990s and early years of this century have seen a series of large-scale mergers and acquisitions in the Pharmaceutical and Biotech arena. These activities each required integration at multiple levels. One of the most important activities is the integration of the R&D pipelines of the participants. We outline the combined portfolio and bioinformatic strategy that was used, and detail the lessons learned for the longer term, from the GlaxoWellcome-SmithKline Beecham merger in 2000. To date, this has been the largest merger of two equally sized Pharma R&D organisations. PMID- 19121409 TI - Kinase-targeted libraries: the design and synthesis of novel, potent, and selective kinase inhibitors. AB - Protein kinases continue to hold tremendous promise for therapeutic intervention, and the search for novel, safe and efficacious kinase inhibitors has intensified over the past decade. Given that most kinases are readily inhibited by organic small molecules and that a wealth of structural data exists on kinase-inhibitor complexes, there has been almost universal success in the design and identification of potent kinase inhibitors. The issues of non-selectivity and congested IP space, however, present formidable challenges for the successful clinical development of these compounds. We describe a systematic approach implemented at Abbott to enable the rapid discovery and design of novel and potent kinase inhibitors that provide additional opportunities for targeting new intellectual property space and achieving acceptable selectivity profiles. PMID- 19121407 TI - Airway smooth muscle as an immunomodulatory cell. AB - Although pivotal in regulating bronchomotor tone in asthma, airway smooth muscle (ASM) also modulates airway inflammation in asthma. ASM myocytes secrete or express a wide array of immunomodulatory mediators in response to extracellular stimuli, and in chronic severe asthma, increases in ASM mass may also render the airway irreversibly obstructed. Although the mechanisms by which ASM secretes cytokines and chemokines are shared with those regulating immune cells, there exist unique ASM signaling pathways that may provide novel therapeutic targets. This review provides an overview of our current understanding of the proliferative as well as synthetic properties of ASM. PMID- 19121410 TI - Novel approaches to the pharmacotherapy of pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) has evolved from an untreatable condition to a disease for which several classes of drugs have now been approved, including various prostanoids, endothelin receptor antagonists and phosphodiesterase-5 inhibitors. Because the pathogenesis of pulmonary hypertension is increasingly understood, various new substances are now under clinical investigation, including serotonin antagonists, vasoactive intestinal peptide, stimulators of soluble guanylate cyclase and tyrosine kinase inhibitors. Several of these compounds hold promise for the future therapy of PAH, especially as regression of pulmonary vascular remodeling appears to become a realistic possibility with the combination of established and novel treatments. PMID- 19121411 TI - G-protein-coupled receptor-focused drug discovery using a target class platform approach. AB - In recent years, several large pharmaceutical companies have taken a novel approach to drug discovery biology and chemistry in that they channel their efforts with respect to particular target classes, such as G-protein-coupled receptors (GPCRs), toward dedicated, specialized teams. Benefits of such an organizational structure are the prospects of establishing several target-family specific experimental techniques and skill sets, thereby enabling a comprehensive functional profiling of drug candidates in different pharmacological respects. In this context, the recently increased number of reports on GPCR ligand-biased signaling has further spurred the efforts in the pharmaceutical industry toward broader biological characterization of the test compounds, for example employing high-content screening to analyze different GPCR ligand-induced signaling pathways. The knowledge of the disease-relevant functional properties of the small molecule GPCR ligands enables target-specific chemical optimization and GPCR-subclass-directed library design. In the case of GPCRs, where little- although at present slowly expanding--structural information on the targets is available, the modeling of GPCR structures crucially depends on biological validation (typically supported by site-directed mutagenesis of the GPCR ligand binding site). In this review, we aim to recapitulate efforts in the pharmaceutical industry to address GPCR-directed drug discovery in a target-class directed platform approach: establishing GPCR-specific biological assay panels and creating computational chemistry methods for finding and optimizing small molecules modulating the activity of GPCRs. PMID- 19121412 TI - Hepatic cyst? PMID- 19121413 TI - Multidisciplinary approach to diagnosis and management of intraductal papillary mucinous neoplasms of the pancreas. AB - Intraductal papillary mucinous neoplasms have gained recognition in recent years as premalignant precursors to pancreatic cancer that enable early detection and often are found incidentally at imaging. Accurate diagnosis and optimal, finely tuned management of these lesions are important and require collaboration across various disciplines, including radiology, endoscopy, surgery, and pathology. Several imaging modalities can visualize these lesions adequately, each with specific advantages and disadvantages. Multidetector computed tomography and magnetic resonance cholangiopancreatography are generally the first-line imaging modalities; endoscopic imaging such as endoscopic ultrasound and endoscopic retrograde cholangiopancreatography are beneficial when the former 2 modalities are equivocal. Surgical candidates generally include patients with main duct lesions or branch duct lesions greater than 3 cm or any possessing a solid component. A management algorithm indicating when surgery should be pursued is proposed. For nonsurgical and postsurgical patients, follow-up management is important to monitor growth and recurrence, and risks from repeated radiation exposure should be taken into account. Furthermore, issues of multifocality and increased predisposition of the pancreas to ductal adenocarcinoma must be addressed at follow-up evaluation. A follow-up management algorithm also is proposed in this review. PMID- 19121414 TI - Strain-specific immunity may drive adaptive polymorphism in the merozoite surface protein 1 of the rodent malaria parasite Plasmodium chabaudi. AB - Clinical immunity against malaria is slow to develop, poorly understood and strongly strain-specific. Understanding how strain-specific immunity develops and identifying the parasite antigens involved is crucial to developing effective vaccines against the disease. In previous experiments we have shown that strain specific protective immunity (SSPI) exists between genetically distinct strains (cloned lines) of the rodent malaria parasite Plasmodium chabaudi chabaudi in mice [Cheesman, S., Raza, A., Carter, R., 2006. Mixed strain infections and strain-specific protective immunity in the rodent malaria parasite P. chabaudi chabaudi in mice. Infect. Immun. 74, 2996-3001]. In two subsequent studies, we identified the highly polymorphic Merozoite Surface Protein 1 (MSP-1) as being the principal candidate molecule for the control of SSPI against P. c. chabaudi malaria [Martinelli et al., 2005; Pattaradilokrat, S., Cheesman, S.J., Carter R., 2007. Linkage group selection: towards identifying genes controlling strain specific protective immunity in malaria. PLoS ONE 2(9):e857]. In the present study, we sequenced the whole msp1 gene of several genetically distinct strains of P. chabaudi and found high levels of genetic diversity. Protein sequence alignments reveal extensive allelic polymorphism between the P. chabaudi strains, concentrated primarily within five regions of the protein. The 3'-end sequence region, encoding the C-terminal 21 kDa region (MSP-1(21)), which is analogous and homologous to MSP-1(19) of Plasmodium falciparum, appears to have been subject to balancing selection. We have found that the strains with the lowest sequence identity at MSP-1(21) (i.e. AS/CB and AJ/CB) induce robust and reciprocal SSPI in experimental mice. In contrast, two strains that do not induce reciprocal SSPI are identical at the 21 kDa region. Final identification of the region(s) controlling SSPI will provide important information to help guide decisions about MSP-1 based vaccines. PMID- 19121416 TI - Bone structure and remodelling in stroke patients: early effects of zoledronate. AB - INTRODUCTION: We have reported that after an acute stroke, intravenous zoledronate prevented bone loss in the hemiplegic hip. Participants from the trial also volunteered for trans-iliac bone biopsy, to assess the early effects of stroke and zoledronate on iliac bone remodelling. METHODS: Patients with acute stroke were randomly assigned to a single intravenous dose of zoledronate 4 mg or placebo within 5 weeks of stroke. Biopsies from 14 patients (3 female, 11 male, mean age 71+/-11) were suitable for analysis. These were taken at mean 10 weeks (+/-2) post-stroke, and included 5 patients who had received zoledronate. Histomorphometry was performed on undecalcified sections using light and fluorescence microscopy. Static and dynamic indices of remodelling were compared to a local reference range from healthy controls. Osteoclasts and their precursors were identified on frozen sections using tartrate resistant acid phosphatase (TRAP) staining. Dual-energy x-ray absorptiometry (DXA) of the proximal femora was performed at baseline and 6 months later. RESULTS: The eroded surface in cancellous bone (ES/BS) was significantly higher in stroke patients than controls (5.7% vs. ref 1.6%, p<0.0001). Although ES/BS did not differ between zoledronate and placebo-treated groups, there were significantly fewer osteoclasts and their precursors in zoledronate-treated individuals (p=0.023). Bone formation indices (osteoid surface, OS/BS and mineralising surface, MS/BS) were significantly lower in stroke patients than controls and although OS/BS was higher in the zoledronate group than the placebo group (p=0.033), MS/BS was not different (p=0.924). There were no differences between hemiplegic and unaffected sides for any histomorphometric parameter despite asymmetric reductions in hip bone mineral density (p=0.013). CONCLUSION: Stroke patients had higher resorption indices and lower bone forming surfaces than controls, consistent with uncoupling of bone remodelling. These findings are preliminary and a larger study is required to evaluate the contributions of gender, age and hemiplegic status to the remodelling imbalance. Zoledronate therapy was associated with a reduction in osteoclastic cell numbers consistent with its known mode of action in bone. PMID- 19121417 TI - Leveraging the purchasing power of the staff nurse. PMID- 19121415 TI - Heat-shock proteins can promote as well as regulate autoimmunity. AB - Heat-shock proteins (Hsps) are among the most highly conserved and immunogenic proteins shared by microbial agents and mammals. Under physiological conditions, the ubiquitously distributed Hsps maintain the integrity and function of other cellular proteins when cells are exposed to stressful stimuli. However, owing to their conserved nature and stress inducibility, Hsps may become targets of immune response. The T cells and/or antibodies induced by a microbial Hsp may crossreact with the corresponding mammalian Hsp (molecular mimicry) and trigger an autoimmune response, which if unchecked can lead to immune pathology and clinical manifestations. Furthermore, enhanced expression of Hsp under stress can unveil previously hidden antigenic determinants that can initiate and perpetuate autoimmune reactivity. Also, the innate immune mechanisms activated by an Hsp can reinforce and even direct the type of adaptive immune response to that protein. Hsps have been implicated in the induction and propagation of autoimmunity in several diseases, including rheumatoid arthritis, atherosclerosis and type 1 diabetes. However, Hsps possess immunoregulatory attributes as well and therefore, are being exploited for immunomodulation of various immune-mediated disorders. PMID- 19121419 TI - The future of nursing. PMID- 19121420 TI - Construction of an integrated surgical suite in a military treatment facility. AB - The OR has long existed as a highly specialized and distinct entity within a hospital. Recent technological advances have allowed the OR to be seamlessly integrated with the larger hospital and, in a virtual sense, with the world. The modern, integrated OR can enhance patients' surgical experiences, improve surgeons' control of OR technology, and increase OR efficiency by reducing surgical times. The Naval Medical Center in San Diego, California, is one of the first military health care systems in the southwest to build a fully integrated, voice-controlled OR. The renovation project transformed 12 ORs into fully integrated surgical suites. PMID- 19121421 TI - Body piercing among Brazilian surgical patients. AB - The practice of body piercing is becoming more common worldwide. Surgical patients with body piercings require special preoperative assessment to ensure safety during and after surgery. In a study of Brazilian surgical patients at one facility, only 0.6% had body piercings. An RN or nursing student instructed each patient on piercing removal, and piercings were removed by the patients before surgery. PMID- 19121422 TI - Perceived effects of specialty nurse certification: a review of the literature. AB - Recent evidence suggests that specialty nurse education and certification may improve the quality of patient care. Specialty nurse certification also may improve nurses' job satisfaction and sense of empowerment, as well as positively affect collaboration with other health care team members. Despite the evidence that there are intrinsic rewards for specialty certification, the lack of extrinsic value to nurses makes it unlikely that greater numbers of nurses will be attracted to certification unless health care administrators increase opportunities for recognition and greater compensation. PMID- 19121423 TI - Volunteering speaks to the heart of nursing. PMID- 19121424 TI - Perioperative fluid management. AB - Technological advances have changed how traditional surgical procedures are performed. New knowledge and surgical skills are required to effectively monitor the patient and manage fluids administered perioperatively. Today, selective fluids are used in a variety of surgical specialties. Complications of fluid therapy can occur that are intrinsic to each procedure within a specialty. This article provides perioperative nurses with the information needed to evaluate different fluid media and their applications and identify patient care considerations. Possible complications and important postanesthesia care concerns also are discussed. PMID- 19121425 TI - Anticoagulants: be alert for errors. PMID- 19121426 TI - Do-not-resuscitate. Orders in surgery: decreasing the confusion. AB - Patients have the right to make decisions regarding their medical care, including the right to refuse treatment or to issue do-not-resuscitate orders as part of an advance directive. Health care providers must comply with the patient's wishes regarding care. Automatic suspension or continuation of a do-not-resuscitate order for a patient undergoing surgery cannot be justified. Surgical team members should consult with the patient and, if necessary, with an ethics expert or committee to determine whether the do-not-resuscitate order is to be maintained or completely or partially suspended during anesthesia and surgery. All surgical departments should have a written policy and procedure concerning the treatment of patients with do-not-resuscitate orders. PMID- 19121427 TI - Relation of clinically defined spontaneous reperfusion to outcome in ST-elevation myocardial infarction. AB - In patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI), a patent infarct-related artery on initial angiography was associated with better angiographic results and improved prognosis compared with patients without spontaneous reflow. Little is known about the prevalence, clinical course, and optimal management of patients presenting with clinical signs of spontaneous reperfusion (SR). The objective was to evaluate characteristics and clinical outcomes in patients with STEMI with clinical signs of SR. The study included 710 consecutive patients with STEMI eligible for reperfusion therapy who were followed up for 30 days. SR was defined as a >or=70% reduction in sum ST elevation and pain severity before initiation of reperfusion therapy. SR was observed in 155 patients (22%). Although almost all patients with STEMI without SR underwent primary reperfusion using primary PCI (398 of 555 patients; 72%) or thrombolysis (125 of 555; 23%), most patients with SR were initially treated conservatively, and primary PCI was performed in only 13 patients (8%). Although patients with SR had a higher incidence of recurrent in-hospital ischemia, they developed smaller myocardial infarctions and sustained less in-hospital cardiogenic shock, heart failure, and electrical complications and had lower 7- and 30-day mortality rates. On multivariate analysis, SR remained significantly associated with a lower incidence of the combined end point of 30-day mortality, congestive heart failure, and recurrent acute coronary syndrome. In conclusion, despite initial conservative therapy, the outcome of patients with SR was markedly better than for patients without SR who underwent primary reperfusion. PMID- 19121428 TI - ST resolution 1 hour after fibrinolysis for prediction of myocardial infarct size: insights from ASSENT 3. AB - Acute ST-segment elevation myocardial infarction requires prompt restoration of myocardial perfusion to salvage myocardium at risk of ischemic necrosis and improve clinical outcome. Early resolution of ST-segment elevation during the time after reperfusion has been associated with both these end points. From the ASsessment of the Safety and Efficacy of a New Thrombolytic regimen (ASSENT) 3 trial, 3,425 patients were analyzed to investigate whether the amount of ST segment resolution, divided into 3 groups (complete, >70%; partial, 30% to 70%; and no resolution, <30%), in the first hour after initiation of therapy was a predictor of final infarct size, estimated by peak creatine kinase and Selvester QRS score on the discharge electrocardiogram. Complete compared with partial and no ST resolution resulted in significantly (p<0.001) smaller infarct sizes of 10.5%, 13.2%, and 15.0% of the left ventricle and significantly (p=0.001) fewer patients with peak creatine >5 times the upper reference level at 50.3%, 71.8%, and 76.3%, respectively. In conclusion, our findings supported previous smaller studies suggesting that early resolution of ST elevation, as a sign of early myocardial reperfusion, resulted in less myocardial damage and preservation of left ventricular function. PMID- 19121429 TI - Usefulness of three posterior chest leads for the detection of posterior wall acute myocardial infarction. AB - A significant proportion of patients with myocardial infarction are missed upon initial presentation to the emergency department. The 12-lead electrocardiogram (ECG) has a low sensitivity for the detection of acute myocardial infarction, especially if the culprit lesion is in the left circumflex artery (LCA). This study was designed to evaluate the benefit of adding 3 posterior chest leads on top of the 12-lead ECG to detect ischemia resulting from LC disease, using a model of temporary balloon occlusion to produce ischemia. We studied 53 consecutive patients who underwent clinically indicated coronary interventions. At the time of coronary angiography, the balloon was inflated to produce complete occlusion of the proximal LCA. We recorded and analyzed the changes noted on the 15-lead ECG, which included 3 posterior leads in addition to the standard 12 leads. In response to acute occlusion of the LCA, the posterior chest leads showed more ST elevation than the other leads, and more patients had ST elevation in the posterior leads than in any other lead. The 15-lead ECG was able to detect>or=0.5 mm (74% vs 38%, p<0.0001) and >or=1 mm (62% vs 34%, p<0.0001) ST elevation in any 2 contiguous leads more frequently than the 12-lead ECG. In conclusion, the 15-lead ECG identified more patients with posterior myocardial wall ischemia because of temporary balloon occlusion of the LC than the 12-lead ECG. This information may enhance the detection of posterior MI in the emergency department and potentially facilitate early institution of reperfusion therapy. PMID- 19121430 TI - Effect of previous treatment with statins on outcome of patients with ST-segment elevation myocardial infarction treated with primary percutaneous coronary intervention. AB - Beyond lipid-lowering effects, statins have favorable effects on platelets, endothelial function, plaque stability, and inflammation. These "pleiotropic" effects could contribute to microvascular function preservation during ischemia. Data are limited about the impact of previous treatment with statins on outcomes of patients with ST-elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI). Accordingly, the aim was to evaluate the effect of previous statin treatment on clinical outcomes of such patients. A total of 950 consecutive patients with STEMI treated with primary PCI who were included in our primary PCI registry from January 2001 to July 2007 were studied. Excluded were patients with cardiogenic shock. Patients were allocated into 2 groups: those who received previous statin treatment (n=327) and those who did not (n=623). Patients who received previous statin treatment were older and more likely to be women; have diabetes, hypertension, hyperlipidemia, renal insufficiency, and anemia; or have had a previous myocardial infarction. Procedural characteristics were similar between the 2 groups. Despite the higher risk profile, patients who received previous statin treatment had a lower 30-day mortality rate (1.5% vs 3.8%; p=0.05). However, at 6 months, mortality differences were no longer evident and patients who received previous statin therapy had a higher rate of target-vessel revascularization (12.4% vs 7.6%; p=0.02). Multivariate analysis showed that previous statin treatment was associated with an odds ratio of 0.4 (95% confidence interval 0.13 to 0.96, p=0.045) for 30-day mortality. In conclusion, the present study suggested that previous therapy with statins in patients with STEMI treated using primary PCI may be associated with reduced short-term mortality. PMID- 19121431 TI - Outcomes and quality of life in patients>or=85 years of age with ST-elevation myocardial infarction. AB - The oldest old comprise the fastest growing segment of the US population. However, data are limited regarding the treatment and outcomes of ST-segment elevation myocardial infarction (STEMI) in this age group. We analyzed consecutive patients with STEMI>or=85 years old at a single center. Quality of life was assessed using the EQ-5D Index (range -0.11 to 1.00) and EQ-VAS (range 0 to 100). Of 1,847 patients admitted from 2002 to 2007 with STEMI, 73 (4%) were >or=85 years old (range 85 to 94). Median time from symptom onset to hospital arrival was 3 hours. Cardiogenic shock occurred in 33%. Primary percutaneous coronary intervention (PCI) was performed in 70% of patients, and the procedural success rate was 94%. Evidenced-based therapy included aspirin (97%), clopidogrel (93%), beta blockers (82%), angiotensin-converting enzyme inhibitors/angiotensin receptor blockers (74%), and statins (86%). The in-hospital mortality rate was 32%, and it was 54% in those with cardiogenic shock. Long-term follow-up was obtained in 96% of hospital survivors at a median of 429 days. Survival rates in patients discharged alive were 75% at 1 year and 65% at 2 years. Cardiogenic shock was the only independent predictor of in-hospital mortality (odds ratio 3.8, 95% confidence interval 1.2 to 11.7, p=0.02), and primary PCI was the only independent predictor of long-term survival (hazard ratio 0.3, 95% confidence interval 0.1 to 0.8, p=0.02). Mean EQ-5D Index was 0.78 and mean EQ-VAS was 70.5. In conclusion, in the oldest old with STEMI, aggressive treatment is associated with reasonable long-term survival and excellent quality of life. The exception may be patients presenting with cardiogenic shock, for whom short-term mortality remains exceedingly high. PMID- 19121432 TI - Thrombocytopenia in patients with an acute coronary syndrome (from the Global Registry of Acute Coronary Events [GRACE]). AB - The incidence of thrombocytopenia after hospital admission, patient and treatment characteristics, and outcomes in patients enrolled in the prospective multinational GRACE were examined. Heparin (unfractionated or low molecular weight) and glycoprotein IIb/IIIa-inhibition can be associated with immune mediated thrombocytopenia of clinical importance. The prevalence of thrombocytopenia in patients with acute coronary syndromes (ACSs) in general and specifically related to these therapies and associated outcomes have been studied little outside of clinical trials. Patients with an ACS were stratified into 4 groups of those with heparin-induced thrombocytopenia (HIT), those with glycoprotein IIb/IIIa-associated thrombocytopenia (GAT), those with other thrombocytopenia (not diagnosed as HIT or associated with glycoprotein inhibitors), and those with no thrombocytopenia. From June 2000 to September 2007, a total of 52,647 patients with an ACS and information for platelet count were enrolled in GRACE. Of these, 152 (0.3%) were reported to develop HIT, 324 (0.6%) developed GAT, and 368 (0.7%) developed other thrombocytopenia. Patients with HIT, GAT, or other thrombocytopenia were significantly more likely to die in the hospital versus those without these diseases (adjusted odds ratio [OR] 1.94, 95% confidence interval [CI] 1.07 to 3.53; adjusted OR 3.45, 95% CI 2.35 to 5.05; and adjusted OR 2.83, 95% CI 1.97 to 4.06, respectively). They were also more likely to experience major bleeding, (re)infarction, or stroke. In conclusion, in this large multinational registry, 1.6% of patients with ACS were reported to develop thrombocytopenia, with only 0.3% being HIT. Regardless of whether patients had clinically recognized HIT, GAT, or other thrombocytopenia, all 3 groups had significantly higher rates of major bleeding, recurrent infarction, stroke, and death. PMID- 19121433 TI - Comparison of six-month outcomes for primary percutaneous revascularization for acute myocardial infarction with drug-eluting versus bare metal stents (from the APEX-AMI study). AB - We evaluated the use and outcomes of drug-eluting stents (DESs) and bare metal stents (BMSs) in a large primary percutaneous coronary intervention (PCI) acute ST-elevation myocardial infarction (MI) trial. Recently concerns have been raised with "off-label" use of DESs for short- and long-term clinical outcomes. Limited randomized data exist evaluating DESs versus BMSs in ST-elevation MI. Patients (n=5,745) in the Assessment of Pexelizumab in Acute Myocardial Infarction (APEX AMI) trial were categorized by stent type used. Baseline variables and clinical outcomes were collected at 90 days and 6 months. Outcomes by stent type were adjusted for using conventional multivariable predictors of 90-day mortality (age, anterior location, total ST-segment deviation, and Killip class), time to PCI, and Thrombolysis In Myocardial Infarction grade flow. Stents were deployed (at the investigator's discretion) in 5,124 patients (89.2%) with acute MI, with DES use in 2,221 (43.3%) and BMS use in 2,903 (56.7%). Patients receiving DESs were younger (median 59 vs 63 years of age, p<0.001), had left anterior descending coronary artery PCI (57.9% vs 48.1%, p<0.001), and often were treated in the United States (58.2%). DES-treated patients had a lower adjusted mortality at 90 days (hazard ratio 0.73, 95% confidence interval [CI] 0.54 to 0.99, p=0.046) and trended toward lower mortality (hazard ratio 0.77, 95% CI 0.58 to 1.03, p=0.084) and recurrent MI (hazard ratio 0.81, 95% CI 0.59 to 1.11, p=0.186) at 6 months compared with BMSs. In conclusion, in this observational analysis of stent use from a large primary percutaneous intervention for acute MI trial, DESs appear as safe as BMSs with similar 6-month clinical outcomes with regard to death and recurrent MI. PMID- 19121434 TI - Comparison of drug-eluting stents and bare-metal stents for the treatment of unprotected left main coronary artery disease in acute coronary syndromes. AB - The aim of this study was to compare long term clinical outcome after left main stenting in a large real world population of patients treated with drug-eluting stents (DES) or bare-metal stents (BMS) in the setting of acute coronary syndromes. The advent of DES decreased the risk of unprotected left main coronary artery restenosis as compared with BMS, but safety concerns still exist, especially when high-risk patients presenting with acute coronary syndromes are considered. The Gruppo Italiano Studi Emodinamici-Societa' Italiana di Cardiologia Invasiva (GISE-SICI) registry is a retrospective, observational multicenter registry promoted by the Italian Society of Invasive Cardiology in which 19 high-volume participating centers enrolled 1,453 consecutive patients who underwent percutaneous coronary intervention on unprotected left main coronary artery between January 2002 and December 2006. From the registry, a total of 849 consecutive patients presenting with unstable angina or non-ST segment elevation myocardial infarction who underwent DES (n=611) or BMS (n=238) implantation were analyzed with extensive multivariable and propensity-score adjustments. At 3-year follow-up, the adjusted hazard ratio for the risk of mortality after DES implantation relative to BMS implantation was 0.90 (95% confidence intervals [CI] 0.59 to 1.38, p=0.617), and the adjusted hazard ratio for the risk of cardiac mortality was 0.75 (95% CI 0.45 to 1.27, p=0.287). DES were associated with significant reduction of myocardial infarction (hazard ratio 0.37, 95% CI 0.17 to 0.81, p=0.009) and target lesion revascularization rates (hazard ratio 0.38, 95% CI 0.21 to 0.69, p=0.001). Younger age, low ejection fraction, an increase in the cardiac biomarkers, absence of diabetes, and bifurcations showed a significant interaction with assigned treatment regarding myocardial infarction. The treatment effects on target lesion revascularization were consistent across multiple subgroups, with the possible exception of patients with nonbifurcational lesions. In conclusion, in a large population of patients with acute coronary syndromes and unprotected left main coronary artery disease DES were more effective than BMS in reducing myocardial infarction and target lesion revascularization. There was no evidence of a significant reduction in mortality with DES versus BMS. PMID- 19121435 TI - Comparison of long-term efficacy of the paclitaxel-eluting stent versus the bare metal stent for treatment of unprotected left main coronary artery disease. AB - The use of paclitaxel-eluting stents (PES) for the treatment of unprotected left main coronary artery (LMCA) disease is controversial. Between January 2003 and December 2006, a total of 287 patients undergoing percutaneous coronary intervention for LMCA lesions were consecutively registered. Of those patients, 178 received PES and 109 received bare-metal stents (BMS). Estimated perioperative mortality rates were 7.3% and 6.8% for the BMS and PES groups, respectively (p=0.51). PES recipients had distal left main bifurcation lesions more frequently compared with BMS recipients (72 vs 42%, p<0.01). At an average follow-up of 35 months, the rates of major adverse cardiac events (4.5 vs 23.9%, adjusted odds ratio [OR] 0.23, 95% confidence interval [CI] 0.09 to 0.58, p<0.001) and target-lesion revascularization (2.2 vs 13.8%, adjusted OR 0.26, 95% CI 0.08 to 0.83, p<0.001) were significantly lower in the PES group than in the BMS group. Overall thrombotic event rates were 1.1% and 4.6% in the PES and BMS groups, respectively (p=0.08). Angiographic follow-up was performed in 61% and 59% of PES and BMS recipients, respectively. The angiographic restenosis rate was significantly lower in the PES group as compared with the BMS group (3.7 vs 23.4%, p<0.001). In conclusion, PES implantation provides a safe, effective therapy for unprotected LMCA disease and decreases the risk of major adverse cardiac events compared with BMS at a mean follow-up of 35 months. PMID- 19121436 TI - Paclitaxel-eluting stent long-term outcomes in percutaneous saphenous vein graft interventions (PELOPS) study. AB - This study examined the 1-year clinical and angiographic follow-up results of implantation of paclitaxel-eluting stents (PES) in aortocoronary saphenous vein graft (SVG) lesions. Sixty-eight consecutive patients with 90 nonoccluded SVG lesions were treated with PES (Taxus), size ranging from 2.25 to 4.5 mm. Angiographic follow-up was performed on 63 patients (93%) and 83 lesions (92%) at 12 months; major adverse cardiac event (MACE) was recorded in all patients at 1 year. The mean age of patients was 71+/-8 years with predominance of men (75%); the mean graft age was 13+/-4 years. Glycoprotein IIb/ IIIa inhibitors were given in 21 patients (31%); embolic protection devices were used in 54 lesions (60%). On average, patients received 1.4 stents per lesion with a stent size of 3.4+/ 0.6 mm and a length of 35.8+/-27.0 mm. Angiographic follow-up revealed a late loss of 0.36+/-0.66 mm with an in-segment binary restenosis rate of 7%. The in hospital MACE was 7%, which was solely contributed by 5 patients with postprocedure non-Q myocardial infarction; the 1-year MACE was 15%, accounted by 1 noncardiac death and 9 patients with target vessel revascularization. Peripheral vascular disease and the use of glycoprotein IIb/ IIIa inhibitors were the independent predictors of MACE at 1 year. In conclusion, implantation of PES to treat degenerative aortocoronary SVG lesions is safe and associated with low late loss, angiographic restenosis, and MACE at 1-year follow-up. PMID- 19121437 TI - Three-year survival following multivessel percutaneous coronary intervention with bare-metal or drug-eluting stents in unselected patients. AB - Drug-eluting stents (DESs) have been shown to reduce the rate of repeat revascularization compared with bare-metal stents (BMSs) after multivessel percutaneous coronary intervention in carefully selected patients. However, the long-term safety and efficacy of DESs in patients with multivessel disease outside the setting of randomized trials was unknown. Therefore, all patients undergoing multivessel percutaneous coronary intervention with BMSs, sirolimus eluting stents (SESs), or paclitaxel-eluting stents (PESs) from January 2000 to December 2005 were investigated. The primary end point was all-cause mortality. A total of 1,720 patients were recruited in 3 consecutive sequential groups of BMS (n=701; January 2000 to April 2002), SES (n=293; April 2002 to February 2003), and PES (n=726; February 2003 to December 2005). Overall median follow-up was 1,440 days. There was improved 3-year survival in the SES group (93.7%) compared with both the BMS (86.1%) and PES groups (87.3%), which remained significant after propensity score adjustment for differences in baseline and procedural characteristics (SES vs BMS, adjusted hazard ratio 0.53, 95% confidence interval 0.30 to 0.94; SES vs PES, adjusted hazard ratio 0.49, 95% confidence interval 0.28 to 0.87). There was no difference in mortality between the PES and BMS groups. Both DES types significantly reduced the need for clinically driven target-vessel and target-lesion revascularization without an excess in myocardial infarction or stent thrombosis. In conclusion, both SESs and PESs significantly reduced the need for repeated revascularization in these patients with no excess in mortality. SESs might reduce mortality in patients undergoing multivessel percutaneous coronary intervention. PMID- 19121438 TI - Mechanism of paradoxical ventricular septal motion after coronary artery bypass grafting. AB - Paradoxical septal motion is commonly noted on echocardiography after coronary artery bypass grafting (CABG), but its mechanism is unclear. Cardiac magnetic resonance imaging was performed before and 3 months after CABG in 23 patients. On a mid-left ventricular short-axis cine image, the motion of myocardial landmarks during the cardiac cycle was ascertained relative to a stationary anterior reference point. Before CABG, the movement of the ventricular septum in systole was either posterior or neutral (median -2 mm) in 19 patients, whereas after CABG, the septum moved anteriorly in all 23 patients (+4 mm; p<0.001). (A positive sign indicates anterior motion in ventricular systole, and a negative sign denotes posterior motion.) The motion of the right ventricular free wall was reduced after CABG (-5 vs -3 mm; p=0.002), whereas anterior movement of the lateral left ventricular wall in systole increased (+4 vs +9 mm; p<0.001). There was a positive correlation between degree of anterior movement of the ventricular septum and right ventricular ejection fraction (r=0.47, p=0.023). In conclusion, after CABG, the entire left ventricle translocated anteriorly in systole. Despite preserved right ventricular function, there was restricted motion of the right ventricular free wall suggestive of postoperative adhesions. The pattern of movement observed offers a sound explanation for postoperative paradoxical septal motion. PMID- 19121439 TI - Relation of preoperative radial artery flow-mediated dilatation to nitric oxide bioavailability in radial artery grafts used in off-pump coronary artery bypass grafting. AB - The radial artery is prone to vasospasm after coronary bypass surgery, and endothelial dysfunction is likely to be a key factor. We investigated whether endothelial dysfunction in radial artery conduits is present, and can be identified, preoperatively using a simple noninvasive ultrasound test of radial artery endothelial response, flow-mediated dilatation (FMD). The study population consisted of 126 patients scheduled for coronary artery bypass grafting. The afternoon before operation, patients had noninvasive ultrasound assessment of endothelial function in the left radial artery by FMD, which measures change in arterial size after an increase in flow-an endothelial-dependent response. Surplus graft segments were obtained at operation and nitric oxide bioavailability within the vessels determined from ex vivo responses to acetylcholine. Preoperative FMD in the radial artery was associated with vasorelaxations to acetylcholine in radial artery grafts (p<0.001 for both dose response curves and maximum relaxations), although there was weak borderline association between FMD and vasorelaxations of saphenous vein grafts (p=0.07 for dose-response curves and p<0.05 for maximum relaxations). In multivariate analysis including cardiac risk factors, FMD was a predictor of vasorelaxations of radial artery grafts (beta=0.020, SE=0.009, p=0.030), independent of classic risk factors for atherosclerosis. In conclusion, there is significant interindividual variation in the endothelial function of vessels used for coronary artery bypass surgery, particularly the radial artery. These differences are present and can be identified preoperatively by FMD. PMID- 19121440 TI - On the use of the inverse electrocardiogram leads. AB - This viewpoint expresses reservations about using the inverse (or negative) aVR lead in the so-called Cabrera limb lead sequence and the implementation of inverse representations of any of the other 12 standard electrocardiographic leads. Instead, the author proposes that in patients with suspected acute coronary syndromes and a nondiagnostic electrocardiographic results, electrocardiography be repeated ("double electrocardiography"), with the V1 to V6 electrodes used to record leads V3R, V4R, V7 to V9 to the left of the spine, and V9R to the right of the spine to aid in triaging patients to the appropriate management. PMID- 19121441 TI - Differential use of warfarin for secondary stroke prevention in patients with various types of atrial fibrillation. AB - Anticoagulation therapy significantly reduces the incidence of thromboembolic events in patients with atrial fibrillation (AF), and warfarin therapy at discharge is a class I-indicated drug in patients with ischemic stroke with persistent or paroxysmal AF without contraindications. The aim was to determine whether participation in the Get With The Guidelines-Stroke (GWTG-S) quality improvement program would be associated with improved adherence to anticoagulation guidelines for patients with all types of AF. Adherence to warfarin treatment at hospital discharge was assessed in eligible patients with AF who presented with stroke or transient ischemic attack, based on type of AF. Of patients with stroke, 10.5% presented with some form of AF. When AF was documented using electrocardiography or telemetry (ECG) during the present admission, eligible patients were more likely to receive warfarin compared with patients for whom AF was reported using medical history only (78.8% vs 49.4%; p<0.0001). Improvement after GWTG-S participation in warfarin use was observed in patients with ECG-documented AF (73.8% at baseline vs 88.5% after the intervention; p<0.0001), but not patients using history only. Women and elderly patients were less likely to receive warfarin, and these gaps in treatment did not narrow during the quality improvement program for patients with ECG documented AF and those with history only. In conclusion, anticoagulation for stroke prevention was underused in general for patients with AF, even in such high-risk groups as patients with stroke. GWTG-S was associated with improved adherence for patients with ECG-documented AF, but patients with a history of AF alone were largely untreated. PMID- 19121442 TI - Effect of catheter ablation for isolated paroxysmal atrial fibrillation on longitudinal and circumferential left ventricular systolic function. AB - Isolated paroxysmal atrial fibrillation (AF) is commonly associated with left ventricular (LV) diastolic dysfunction but normal radial systolic contraction. We aim to investigate LV systolic function more precisely using 2-dimensional strain technique in patients with isolated paroxysmal AF and to evaluate evolution of longitudinal, circumferential, and radial (or transverse) strain components after catheter ablation of AF. Thirty patients with isolated paroxysmal AF were investigated by echocardiographic studies before and at 1-day, 1-month, 6-month, and 12-month intervals after radiofrequency ablation. Left heart dimensions and LV systolic and diastolic functions were evaluated at each time interval. LV systolic function was quantified by LV ejection fraction and by 2-dimensional strain evaluation, giving regional and global longitudinal, circumferential, transverse, and radial peak of percentage deformation. Patients with AF were compared with 30 control subjects, paired by age and by sex. Before AF ablation, LV ejection fraction, transverse and radial strains were not significantly different from control subjects. By contrast, global longitudinal and circumferential strains were significantly lower than controls (-17.7%+/-2.4% vs 21.5%+/-2.0% [p<0.01] and -16.0%+/-2.9% vs -20.7%+/-3.4% [p<0.01], respectively). At the end of follow-up, global longitudinal and circumferential strains were significantly improved (-20.8%+/-2.6% vs -17.7%+/-2.4% (p<0.01) and -18.5%+/-3.1% vs -16.0%+/-2.9% [p<0.05], respectively). Global longitudinal strain was not significantly different from normal control subjects at the end of follow-up. In conclusion, this prospective study demonstrates (1) the existence of early longitudinal and circumferential LV systolic function abnormalities in patients with isolated paroxysmal AF but normal ejection fraction and (2) reverse remodeling of these abnormalities after AF ablation. PMID- 19121443 TI - Usefulness of baseline electrocardiographic QRS complex pattern to predict response to cardiac resynchronization. AB - Cardiac resynchronization therapy (CRT) improves morbidity and mortality in patients with heart failure with QRS>120 ms, yet most patients studied in clinical trials manifested baseline left branch bundle block (LBBB). It is unclear whether benefits of CRT extend to patients with right branch bundle block (RBBB) or a paced QRS at baseline despite QRS>120 ms. Orthotopic heart transplantation- and ventricular assist device-free survival, symptomatic response, and echocardiographic response were evaluated in the 636 patients who underwent CRT at our institution from 2000 to 2007 in whom the baseline electrocardiogram showed LBBB (n=412; 65%), paced QRS (n=162; 26%), or RBBB (n=62; 10%). Mortality was assessed using the Social Security Death Index, and the medical record was analyzed for clinical data. A decrease in New York Heart Association class>or=0.5 after >or=6 months of CRT defined symptomatic response. Echocardiographic evidence of improved left ventricular function and reverse remodeling was evaluated after >or=6 months of CRT. Survival free from orthotopic heart transplantation and ventricular assist device placement was best in patients with LBBB and worst in those with RBBB, whereas patients with paced QRS had an intermediate prognosis (p=0.003). This finding remained significant after controlling for baseline differences among the 3 groups. Symptomatic response was observed most often in patients with LBBB (60%), occurred least often in patients with RBBB (14%), and was intermediate in patients with paced QRS (46%; p<0.001). Echocardiographic improvement showed a similar stepwise trend. In conclusion, patients with RBBB undergoing CRT had low rates of symptomatic and echocardiographic response, and their survival free from orthotopic heart transplantation or ventricular assist device placement was significantly worse than in patients with LBBB. Patients with conventionally paced QRS experienced an intermediate response. PMID- 19121444 TI - Relation of brain natriuretic peptide level to extent of left ventricular scarring in patients with chronic heart failure secondary to ischemic cardiomyopathy. AB - Multiple factors influence brain natriuretic peptide (BNP) release in patients with heart failure. We hypothesized that extensive myocardial scarring could result in an attenuated BNP response. A total of 115 patients with New York Heart Association class III chronic heart failure and ischemic cardiomyopathy were evaluated for ischemia, hibernation, and myocardial scarring by dipyridamole rubidium-positron emission tomographic scanning with fluorine-18, 2-fluoro-2 deoxyyglucose. Plasma BNP levels were determined within 2 weeks of the study. Left ventricular dimension and function were evaluated by echocardiography. Patients were categorized as having <33% myocardial scar (n=67) or>or=33% myocardial scar (n=48). BNP measurements were correlated with amount of myocardial scarring. Compared with patients with less scar, those with >or=33% scar had lower BNP levels (mean 317+/-364 vs 635+/-852 pg/ml, median 212 vs 357, p=0.016). Using multiple regression analysis, presence of scarring was associated with decreased BNP response (p=0.022). Further, patients with <33% scar in whom a higher BNP level was noted had more ischemia (51% vs 27%, p=0.01) and greater myocardial hibernation (22+/-14% vs 12+/-7%, p=0.02) compared with patients with >or=33% scar. In conclusion, in patients with chronic heart failure, a decreased BNP response indicated extensive myocardial scarring. PMID- 19121445 TI - Usefulness of hand-carried ultrasound to predict elevated left ventricular filling pressure. AB - Detection of increased left ventricular filling pressure (LVFP) is paramount in the evaluation and treatment of patients with left heart failure, yet difficult to assess directly at the bedside. Although inferior vena cava (IVC) examination can detect increased right atrial pressure, a used surrogate for high LVFP, this assumption has not been directly tested. We evaluated whether assessment of IVC size and respirophasic variation could be used to predict increased LVFP. Seventy two patients with chronic heart failure underwent right heart catheterization and limited echocardiographic examination focused on the IVC. IVC size and collapsibility were evaluated for their ability to predict a wedge pressure>or=15 mm Hg. In conclusion, an IVC maximum dimension of 2.0 cm and IVC collapsibility of 45% were the optimal cutoffs to predict an increased pulmonary capillary wedge pressure, with sensitivities of 75% and 83% and specificities of 83% and 71%, respectively. PMID- 19121446 TI - Comparison of beta blocker and digoxin alone and in combination for management of patients with atrial fibrillation and heart failure. AB - In patients with atrial fibrillation (AF) and heart failure (HF), beta blockers and digoxin reduce the ventricular rate, but controversy exists concerning how these drugs affect prognosis in this setting. This study compared the effects of beta blocker and digoxin on mortality in patients with both AF and HF. In a single-center institution, patients with AF and HF seen between January 2000 and January 2004 were identified and followed until September 2007. Of 1,269 consecutive patients with both AF and HF, 260 were treated with a beta blocker alone, 189 with beta blocker plus digoxin, 402 with digoxin alone, and 418 without beta blocker or digoxin (control group). During a follow-up of 881+/-859 days, 247 patients died. Compared with the control group, treatment with beta blocker was associated with a decreased mortality (relative risk=0.58, 95% confidence interval 0.40 to 0.85, p=0.005 for beta blocker alone and 0.59, 95% confidence interval 0.40 to 0.87, p=0.008 for beta blocker plus digoxin). By contrast, treatment with digoxin alone was not associated with a better survival (relative risk=0.97, 95% confidence interval 0.73 to 1.30, p=NS). Results remained significant after adjustment for potential confounders and similar when we considered, separately, HF with permanent or nonpermanent AF, presence or absence of coronary disease, and patients with decreased or preserved systolic function. In conclusion, in unselected patients with AF and HF, treatments with beta blocker alone or with beta blocker plus digoxin are associated with a similar decrease in the risk of death. Digoxin alone is associated with a worse survival chance, similar to that of patients without any rate control treatment. PMID- 19121447 TI - Practice variation among cardiovascular physicians in management of patients with mitral regurgitation. AB - Severe mitral regurgitation (MR), whether isolated or with coronary disease, was associated with adverse clinical outcomes. However, clinician practice is characterized by different thresholds for treatment. No data exist that described current practice patterns, factors that affected these patterns, or whether physicians followed American College of Cardiology/American Heart Association guidelines in clinical practice for patients with MR. Cardiovascular specialists were surveyed using e-mail, and 1,076 physicians completed the survey (71% response rate), including adult cardiologists (94%) and cardiac surgeons (5%) who practiced in the United States (78%), Canada (5%), and other nations (17%). Wide variations were noted regarding rates of referral of asymptomatic patients with severe MR for mitral valve repair. There was geographic and specialty-dependent heterogeneity in practice. Most physicians (65%) used medications to delay the progression of MR in the absence of guideline recommendations. A minority (28%) of respondents routinely quantitated MR by calculating the effective regurgitant orifice area. In patients undergoing percutaneous coronary intervention, MR severity was not assessed before the procedure by 1 in 4 providers, and the presence of MR frequently did not affect clinical care decisions. In conclusion, considerable variability existed in the clinical management of MR, particularly regarding referral of asymptomatic patients for mitral valve reparative surgery. Medications were frequently used to treat asymptomatic patients with MR in the absence of evidence of pharmacologic efficacy. MR was frequently not considered a relevant factor before treatment of patients with coexistent coronary artery disease. PMID- 19121448 TI - Re-examination of the electrocardiogram in boys with Duchenne muscular dystrophy and correlation with its dilated cardiomyopathy. AB - Duchenne muscular dystrophy (DMD) results in dilated cardiomyopathy (DC). Characteristic electrocardiographic (ECG) changes include short PR interval, right ventricular hypertrophy (RVH), prolonged QTc interval, and prominent Q waves in leads I, aVL, V5, and V6 or in leads II, III, aVF, V5, and V6. We re examined the prevalence and correlation of ECG changes with DC in DMD. Electrograms of 115 patients with DMD were evaluated. DC was defined as an echocardiographic ejection fraction<55%. PR interval and RVH were based on age based normal values. Abnormal Q waves were >or=4 mm. Abnormal QTc interval was >or=450 ms. ST-segment depression was defined as >0.5 mm. Fisher's exact test evaluated significant differences between groups and logistic regression determined whether number of ECG changes predicted DC. Forty had DC. No significant differences existed between the number of ECG changes in DC and non DC groups (p=0.279). Distribution of findings included short PR interval (43%), RVH (37%), prominent Q waves in leads V5 (34%) and V6 (33%), prominent Q waves in leads I, aVL, V5, and V6 (3, 1 with DC), prominent Q waves in leads II, III, aVF, V5, and V6 (9, 4 with DC), long QTc interval (0), ST depression (2, 1 with DC), and flat/biphasic ST segments (38, 15 with DC). In conclusion, ECG changes are similar in patients with DMD regardless of presence of DC. Previously reported characteristic ECG changes are seen in a minority of DMD cases. The most common findings are short PR interval and RVH. Prominent Q waves in leads II, III, aVF, V5, and V6 are more likely. PMID- 19121449 TI - Relation of coarctation of the aorta to the occurrence of ascending aortic dilation in children and young adults with bicuspid aortic valves. AB - Children with bicuspid aortic valve (BAV) have aortic dilation that is present and progressive from birth irrespective of the functional state of the valve. There are no published data comparing aortic dilation in children with BAV with and without aortic coarctation (CoA). The objectives of this study were to (1) compare differences in aortic dimensions and rates of dilation between children with isolated BAV with those of children with BAV and CoA and (2) identify risk factors for the development of aortic dilation. Patients with BAV with CoA (group A) and without CoA (group B) were identified from our echocardiographic database (1993 to 2006). Aortic measurements at 4 levels were obtained, and z scores were compared. Criteria for exclusion were severe aortic regurgitation/stenosis, previous aortic valvuloplasty, complex left-sided cardiac disease, ventricular septal defects, and Turner, Noonan, Williams, and Marfan syndromes. There were 600 echocardiograms in 247 patients. Group A had 192 echocardiograms in 53 patients (median age 11.3 years; range 0 to 30; median follow-up 7 years), and group B had 382 in 194 patients (median age 8.7 years; range 0 to 29; median follow-up 4 years). Group B had significantly greater ascending aorta dimensions (p<0.05) and significantly faster rates of aortic dilation (p<0.0001). The ascending aorta in patients with BAV and CoA did not dilate to the same degree as in patients with isolated BAV. In conclusion, valve morphologic characteristics and function and age at CoA repair had none to minimal impact on aortic dimensions. PMID- 19121450 TI - Prevalence of pericardial effusions in children with large atrial or ventricular septal defect. AB - Recently, a high percentage of adults with isolated secundum atrial septal defects (ASD) and ventricular septal defects (VSD) undergoing surgical repair had a circumferential pericardial effusion (PE), preoperatively. It was our impression that PEs are uncommon in children with unrepaired ASD or VSD. To evaluate this impression, we performed a retrospective study to determine the prevalence of PE in children with ASD compared with those with VSD and with children with normal cardiac anatomy. The preoperative echocardiograms of 90 patients with isolated surgical grade secundum ASDs, 75 patients with isolated VSDs, and 91 normal patients were reviewed to evaluate for the presence or absence of pericardial effusions and also the septal defect size. Only 1 patient (ASD group) of the total 256 had a global PE (0.4%). No large effusions were found in any group. Eighty-one children (32%) had any effusion present in the pericardial space. The prevalence of these small PEs was not significantly different between the ASD (36/90, 40%) and VSD (23/75, 31%, p=0.25) groups. The patients with ASD, however, had a significantly higher prevalence of PEs compared with the control group (22/91, 24%, p<0.03). In conclusion, the presence of global PEs in children with surgical grade ASD or patients with VSD is rare, unlike that reported in adults with the same lesions. Trace effusions are found more commonly in children with ASD than those in VSD or in normal children, which may herald a potential for larger effusions with a longer time before defect closure. PMID- 19121451 TI - Comparison of Nebivolol monotherapy versus Nebivolol in combination with other antihypertensive therapies for the treatment of hypertension. AB - Nebivolol is a novel, beta1-adrenergic receptor blocker with vasodilatory properties mediated through the activation of the L-arginine/nitric oxide pathway. Short-term randomized clinical trials have demonstrated that nebivolol has antihypertensive efficacy benefits comparable to other beta blockers but with a favorable safety and tolerability profile. The long-term safety and efficacy results of oral nebivolol (5, 10, or 20 mg) 1 time/day, as monotherapy or in combination with other antihypertensive agents, were assessed in a double-blind, multicenter, 9-month extension study of 3 phase III, double-blind, 3-month trials in patients with stage I to II hypertension (mean sitting diastolic blood pressures [BPs]>or=95 and 20 years) death rate in initially healthy middle-aged men. AB - The prognostic implications of heart rate (HR) change over years have never been assessed. It was hypothesized that an increase in HR in apparently healthy persons observed over years could be associated with an increase in mortality risk and conversely. A total of 5,139 asymptomatic working men (aged 42 to 53 years) free of clinically detectable cardiovascular disease were recruited from 1967 to 1972 and had their HRs measured at rest in standardized conditions every year for 5 consecutive years. HR change was defined as the difference between HR at examination 5 and HR at inclusion, and subjects were divided into tertiles according to decrease >4 beats/min, unchanged (from -4 to +3 beats/min), and increase >3 beats/min. After >20 years of mortality surveillance, 1,219 deaths were observed. After adjustments were made for confounding factors, including baseline HR at rest, and compared with subjects with unchanged HRs, subjects with decreased HRs during the 5 years had a 14% decreased mortality risk (RR 0.86, 95% confidence interval 0.74 to 1.00, p=0.05), whereas subjects with increased HRs during the 5 years had a 19% increased mortality risk (RR 1.19, and 95% confidence interval 1.04 to 1.37, p<0.012). In conclusion, change in HR at rest over 5 years was an independent predictor of mortality in middle-aged men. PMID- 19121453 TI - Reperfusion strategies in ST-elevation myocardial infarction. PMID- 19121454 TI - Life lessons from modern-day greats in cardiovascular disease. PMID- 19121455 TI - Religion, fasting and coronary artery disease. PMID- 19121456 TI - Management of dyslipidemia associated with protease inhibitors. PMID- 19121457 TI - Diurnal and seasonal patterns in inflammation-sensitive biomarker level. PMID- 19121458 TI - Evidence of early right ventricular and septal mechanical activation. PMID- 19121459 TI - Dr. Samuel levine and mitral stenosis. PMID- 19121460 TI - Submitting manuscripts to biomedical journals: common errors and helpful solutions. PMID- 19121461 TI - Chiropractic management of myofascial trigger points and myofascial pain syndrome: a systematic review of the literature. AB - OBJECTIVES: Myofascial pain syndrome (MPS) and myofascial trigger points (MTrPs) are important aspects of musculoskeletal medicine, including chiropractic. The purpose of this study was to review the most commonly used treatment procedures in chiropractic for MPS and MTrPs. METHODS: The Scientific Commission of the Council on Chiropractic Guidelines and Practice Parameters (CCGPP) was charged with developing literature syntheses, organized by anatomical region, to evaluate and report on the evidence base for chiropractic care. This article is the outcome of this charge. As part of the CCGPP process, preliminary drafts of these articles were posted on the CCGPP Web site www.ccgpp.org (2006-8) to allow for an open process and the broadest possible mechanism for stakeholder input. PubMed, Excerpta Medica Database, Cumulative Index to Nursing and Allied Health Literature, and databases for systematic reviews and clinical guidelines were searched. Separate searches were conducted for (1) manual palpation and algometry, (2) chiropractic and other manual therapies, and (3) other conservative and complementary/alternative therapies. Studies were screened for relevance and rated using the Oxford Scale and Scottish Intercollegiate Guidelines Network rating system. RESULTS: A total of 112 articles were identified. Review of these articles resulted in the following recommendations regarding treatment: Moderately strong evidence supports manipulation and ischemic pressure for immediate pain relief at MTrPs, but only limited evidence exists for long-term pain relief at MTrPs. Evidence supports laser therapy (strong), transcutaneous electrical nerve stimulation, acupuncture, and magnet therapy (all moderate) for MTrPs and MPS, although the duration of relief varies among therapies. Limited evidence supports electrical muscle stimulation, high voltage galvanic stimulation, interferential current, and frequency modulated neural stimulation in the treatment of MTrPs and MPS. Evidence is weak for ultrasound therapy. CONCLUSIONS: Manual-type therapies and some physiologic therapeutic modalities have acceptable evidentiary support in the treatment of MPS and TrPs. PMID- 19121462 TI - Chiropractic management of fibromyalgia syndrome: a systematic review of the literature. AB - OBJECTIVE: Fibromyalgia syndrome (FMS) is one of the most commonly diagnosed nonarticular soft tissue conditions in all fields of musculoskeletal medicine, including chiropractic. The purpose of this study was to perform a comprehensive review of the literature for the most commonly used treatment procedures in chiropractic for FMS and to provide evidence ratings for these procedures. The emphasis of this literature review was on conservative and nonpharmaceutical therapies. METHODS: The Scientific Commission of the Council on Chiropractic Guidelines and Practice Parameters (CCGPP) was charged with developing literature syntheses, organized by anatomical region, to evaluate and report on the evidence base for chiropractic care. This article is the outcome of this charge. As part of the CCGPP process, preliminary drafts of these articles were posted on the CCGPP Web site www.ccgpp.org (2006-8) to allow for an open process and the broadest possible mechanism for stakeholder input. Online comprehensive literature searches were performed of the following databases: Cochrane Database of Systematic Reviews; National Guidelines Clearinghouse; Cochrane Central Register of Controlled Trials; Manual, Alternative, and Natural Therapy Index System; Index to Chiropractic Literature, Cumulative Index to Nursing and Allied Health Literature; Allied and Complementary Medicine; and PubMed up to June 2006. RESULTS: Our search yielded the following results: 8 systematic reviews, 3 meta analyses, 5 published guidelines, and 1 consensus document. Our direct search of the databases for additional randomized trials did not find any chiropractic randomized clinical trials that were not already included in one or more of the systematic reviews/guidelines. The review of the Manual, Alternative, and Natural Therapy Index System and Index to Chiropractic Literature databases yielded an additional 38 articles regarding various nonpharmacologic therapies such as chiropractic, acupuncture, nutritional/herbal supplements, massage, etc. Review of these articles resulted in the following recommendations regarding nonpharmaceutical treatments of FMS. Strong evidence supports aerobic exercise and cognitive behavioral therapy. Moderate evidence supports massage, muscle strength training, acupuncture, and spa therapy (balneotherapy). Limited evidence supports spinal manipulation, movement/body awareness, vitamins, herbs, and dietary modification. CONCLUSIONS: Several nonpharmacologic treatments and manual type therapies have acceptable evidentiary support in the treatment of FMS. PMID- 19121463 TI - Chiropractic management of tendinopathy: a literature synthesis. AB - OBJECTIVE: Chronic tendon pathology is a soft tissue condition commonly seen in chiropractic practice. Tendonitis, tendinosis, and tendinopathy are terms used to describe this clinical entity. The purpose of this article is to review interventions commonly used by doctors of chiropractic when treating tendinopathy. METHODS: The Scientific Commission of the Council on Chiropractic Guidelines and Practice Parameters (CCGPP) was charged with developing literature syntheses, organized by anatomical region, to evaluate and report on the evidence base for chiropractic care. This article is the outcome of this charge. As part of the CCGPP process, preliminary drafts of these articles were posted on the CCGPP Web site www.ccgpp.org (2006-8) to allow for an open process and the broadest possible mechanism for stakeholder input. A literature search was performed using the PubMed; Cumulative Index to Nursing and Allied Health Literature; Index to Chiropractic Literature; Manual, Alternative, and Natural Therapy Index System; National Guidelines Clearinghouse; Database of Abstracts of Reviews of Effects; and Turning Research Into Practice databases. The inclusion criteria were manual therapies, spinal manipulation, mobilization, tendonitis, tendinopathy, tendinosis, cryotherapy, bracing, orthotics, massage, friction massage, transverse friction massage, electrical stimulation, acupuncture, exercise, eccentric exercise, laser, and therapeutic ultrasound. RESULTS: There is evidence that ultrasound therapy provides clinically important improvement in the treatment of calcific tendonitis. There is limited evidence of the benefit of manipulation and mobilization in the treatment of tendinopathy. Limited evidence exists to support the use of supervised exercise, eccentric exercise, friction massage, acupuncture, laser therapy, use of bracing, orthotics, and cryotherapy in the treatment of tendinopathy. CONCLUSION: Chiropractors often provide a number of conservative interventions commonly used to treat tendinopathy. PMID- 19121464 TI - Manipulative therapy for lower extremity conditions: expansion of literature review. AB - OBJECTIVE: The purpose of this study was to conduct a systematic review on manipulative therapy for lower extremity conditions and expand on a previously published literature review. METHODS: The Scientific Commission of the Council on Chiropractic Guidelines and Practice Parameters (CCGPP) was charged with developing literature syntheses, organized by anatomical region, to evaluate and report on the evidence base for chiropractic care. This article is the outcome of this charge. As part of the CCGPP process, preliminary drafts of these articles were posted on the CCGPP Web site www.ccgpp.org (2006-8) to allow for an open process and the broadest possible mechanism for stakeholder input. The Cumulative Index to Nursing and Allied Health Literature; PubMed; Manual, Alternative, and Natural Therapy Index System; Science Direct; and Index to Chiropractic Literature were searched from December 2006 to February 2008. Search terms included chiropractic, osteopathic, orthopedic, or physical therapy and MeSH terms for each region. Inclusion criteria required a diagnosis and manipulative therapy (mobilization and manipulation grades I-V) with or without adjunctive care. Exclusion criteria were pain referred from spinal sites (without diagnosis), referral for surgery, and conditions contraindicated for manipulative therapy. Clinical trials were assessed using a modified Scottish Intercollegiate Guidelines Network ranking system. RESULTS: Of the total 389 citations captured, 39 were determined to be relevant. There is a level of C or limited evidence for manipulative therapy combined with multimodal or exercise therapy for hip osteoarthritis. There is a level of B or fair evidence for manipulative therapy of the knee and/or full kinetic chain, and of the ankle and/or foot, combined with multimodal or exercise therapy for knee osteoarthritis, patellofemoral pain syndrome, and ankle inversion sprain. There is also a level of C or limited evidence for manipulative therapy of the ankle and/or foot combined with multimodal or exercise therapy for plantar fasciitis, metatarsalgia, and hallux limitus/rigidus. There is also a level of I or insufficient evidence for manipulative therapy of the ankle and/or foot combined with multimodal or exercise therapy for hallux abducto valgus. CONCLUSIONS: There are a growing number of peer-reviewed studies of manipulative therapy for lower extremity disorders. PMID- 19121465 TI - Forces applied to the cervical spine during posteroanterior mobilization. AB - OBJECTIVE: There is little information on manual forces applied during cervical mobilization, a common treatment technique. Potential variability of applied forces between therapists and treatment occasions, and factors associated with different force applications are unknown. The purpose of this study is to establish the baseline mechanical properties of cervical spine mobilization and to determine if the applied forces are affected by the characteristics of therapists and mobilized subjects. METHODS: Physiotherapists (n = 116) applied 4 grades of posteroanterior mobilization to the premarked C2 and C7 spinous (central technique) and articular processes (unilateral technique, one right and one left) of 1 of 35 asymptomatic subjects. Techniques were performed in randomized order, and the first one was repeated after 20 minutes. Load cells attached to the treatment table recorded forces in 3 directions. Before mobilization, subjects' spinal stiffness at the C2 and C7 spinous processes was measured using a custom device. Analyses of variance with Bonferroni post hoc tests determined technique and grade differences, intraclass correlation coefficients the reliability between therapists, and linear regression the factors associated with forces. RESULTS: Therapists apply distinct manual forces for different techniques and grades (P < .001). Variability between therapists is high, but intratherapist reliability is good (intraclass correlation coefficient [2,1] for different force parameters, 0.84-0.93). Mean peak forces increase from grades I to IV, ranging from 22 to 92 N for resultant forces. Greater vertical and caudad-cephalad forces are applied to C7 than C2 (P < .01), with higher mediolateral forces during unilateral techniques (P < .001). Male sex of the therapist or the mobilized subject is associated with higher forces, and C2 stiffness, thumb pain and postgraduate training with lower (P < .05). CONCLUSIONS: These results quantify cervical mobilization forces, which will inform future research aimed at improving its application and clinical effectiveness. PMID- 19121466 TI - Effects of 2 physiotherapy programs on pain perception, muscular flexibility, and illness impact in women with fibromyalgia: a pilot study. AB - OBJECTIVE: This study assessed the effect of 2 physiotherapy programs designed to improve flexibility and to reduce the impact of the illness and pain perception in women with the fibromyalgia syndrome (FMS), and compared the effects of the 2 programs in the short and intermediate term. METHODS: Twenty FMS patients were randomly assigned to 2 training groups, one following a program of kinesiotherapy and active muscular stretching and one using techniques of Global Myofascial Physiotherapy, according to the Mezieres method. Both groups met twice a week for 12 weeks, for a total of 150 minutes each week. Flexibility and illness impact were measured by means of a standard test, whereas pain was assessed by means of thumb palpation. Measurements were taken at the beginning and end of the program and 24 weeks after its end. RESULTS: Patients had achieved a statistically significant reduction in the severity of the disease and improved their flexibility level by the end of the program, but had returned to initial values after follow-up. Significant differences were not observed between the 2 treatment groups in the initial values or in the results at the end of the program or after the follow-up, so neither program proved better than the other. CONCLUSION: The FMS patients in this study improved their flexibility level and general well-being using both kinesiotherapy and stretching exercises techniques. PMID- 19121467 TI - Running posture and step length changes immediately after chiropractic treatment in a patient with xeroderma pigmentosum. AB - OBJECTIVE: This case study reports on selected measures of locomotion (running) in a 5-year-old patient with xeroderma pigmentosum after chiropractic care. CLINICAL FEATURE: A 5-year-old female patient (16.4 kg, 99.1 cm) with xeroderma pigmentosum (type A) volunteered to participate in the experiment with the consent of her parents. The patient had well-documented signs of delayed fine motor (eg, difficulty with writing, coloring, cutting) and gross motor control (eg, balance and coordination dysfunction and falling while running), and delayed speech. INTERVENTION AND OUTCOMES: Trunk forward lean angles, step lengths, and hip horizontal translations were assessed by video as the participant ran as fast as possible down a laboratory runway. After chiropractic manipulation (adjustments), the patient reduced the trunk forward lean angle to become more vertical (P = .000). In addition, the patient experienced an increase in step length (P = .031). No significant change in lateral translation was observed after the intervention. CONCLUSION: For this patient with xeroderma pigmentosum, chiropractic manipulation (adjustments) resulted in immediate changes in running performance. Further investigation is needed to examine the effect of chiropractic on locomotion in both symptomatic and asymptomatic patients. PMID- 19121468 TI - Nutrition in kidney disease: introduction. PMID- 19121470 TI - Why are chronic kidney disease patients anorexic and what can be done about it? AB - The progressive decline of glomerular filtration rate in chronic kidney disease patients is associated with a significant reduction in food intake. Approximately one third of chronic dialysis patients complain of a fair or poor appetite and this is related directly to poor patient outcomes. Appetite regulation involves the gastrointestinal tract (ghrelin as an appetite stimulant, and cholecystokinin, glucagon-like peptide-1, and neuropeptide YY as appetite inhibitors); the adipose tissue with leptin, a potent appetite inhibitor; the vagal system; and the brain, which integrates the stimuli in the hypothalamus area. Satiety relies on the melanocortin receptors with serotonin as the main neurotransmitter and is challenged with hunger peptides, namely, neuropeptide Y and agouti-related peptide. In nondialyzed chronic renal failure patients and in maintenance dialysis patients, anorexia is related mainly to the accumulation of unidentified anorexigenic compounds, inflammatory cytokines, and alterations in appetite regulation, such as amino acid imbalance, which increases the transport of free tryptophan across the blood-brain barrier. This creates a hyperserotoninergic state that is prone to low appetite. Treatment of anorexia involves counseling, starting dialysis treatments in uremic chronic kidney disease patients, increasing the dialysis dose, and possibly using appetite stimulants. PMID- 19121469 TI - Why is protein-energy wasting associated with mortality in chronic kidney disease? AB - Observational studies in chronic kidney disease (CKD) populations consistently have shown the strong mortality-predictability of such markers of protein-energy wasting (PEW) as hypoalbuminemia, low serum cholesterol levels, low body mass index, and reduced dietary protein intake. Even though the PEW-mortality association data traditionally are reported mostly in maintenance dialysis patients, emerging studies extend the existence of these associations to predialysis stages of CKD. Paradoxic risk factor patterns (reverse epidemiology) for both obesity and cholesterol recently have been reported in predialysis CKD, underscoring the overwhelming impact of PEW, a short-term killer, on reversing the long-term effect of conventional cardiovascular risk factors. Multiple pathophysiologic mechanisms have been suggested to explain the link between PEW and mortality in CKD, including derangements in muscle, adipose tissue, and the gastrointestinal, hematopoietic, and immune systems; complications related to deficiencies of multiple micronutrients; and the maladaptive activation of the inflammatory cascade. In addition to well-described pathophysiologic mechanisms involved in the higher mortality seen with PEW, we also discuss the potential role of novel factors such as circulating actin, gelsolin, and proinflammatory high-density lipoprotein. Whether PEW is causally related to adverse outcomes in CKD needs to be verified in randomized controlled trials of nutritional interventions. The initiation of major clinical trials targeting nutritional interventions with the goal of improving survival in CKD offer the promise of extending the survival of this vulnerable patient population. PMID- 19121471 TI - Prevention of cardiovascular disease in chronic kidney disease patients. AB - Therapeutic strategies to prevent cardiovascular disease are based on the ability of the intervention to reduce specific risk factors and subsequent end-organ damage, implying that these factors are causally involved in the pathogenesis of the disease. In subjects with normal kidney function, the control of classic, Framingham-type, risk factors (blood pressure, glycemia, lipids, smoking) reduces impressively the burden of cardiovascular disease. In chronic kidney disease (CKD), prevention is based on similar strategies in general and on reduction of proteinuria (blockade of the renin-angiotensin-aldosterone system [RAAS], salt restriction) in particular. Although several nontraditional risk factors are recognized already in early stages of CKD (stages 1 and 2), they become more prominent in stages 3 to 5, most likely responsible for a different pathology. In fact, cardiovascular and all-cause mortality increase dramatically from CKD stage 3b (glomerular filtration rate < 45 mL/min/1.72 m2) to stage 5, and the pattern of cardiac and vascular damage changes profoundly. It appears that arteriosclerosis is recognized more often than atherosclerosis, and left ventricular hypertrophy or cardiac fibrosis is prominent. Randomized controlled trials in advanced CKD and stage 5D using strategies targeting classic risk factors indicate that treatments are not as effective as in subjects with normal kidney function. Whether late stages of CKD or a risk pattern dominated by nonclassic risk factors need a different or an additional approach or whether we can use biomarkers to detect the turning point is currently a matter of intense discussion among nephrologists and clinical scientists. PMID- 19121472 TI - Do low-protein diets work in chronic kidney disease patients? AB - Patients with chronic kidney disease have been advised for many years to reduce their protein intake. This review addresses the biochemical, pathophysiologic, and nutritional background that underlies this recommendation. The clinical and therapeutic evidence for prescribing such diets is addressed, as well as the potential caveats. A proposed method for managing and monitoring patients also is provided. PMID- 19121473 TI - Causes and prevention of protein-energy wasting in chronic kidney failure. AB - Protein-energy wasting (PEW), defined as reduced somatic and/or circulating body protein mass, decreased fat mass, and usually reduced protein and energy intake, has a prevalence that is variously estimated to be 18% to 75% in maintenance hemodialysis and chronic peritoneal dialysis patients. PEW is associated with increased morbidity and mortality and often is preventable or treatable. Thus, it has been argued that maintenance hemodialysis and chronic peritoneal dialysis patients should be monitored routinely for PEW and treated for this condition, when it occurs. A trend toward PEW can emerge in early stage 3 chronic kidney disease with an increasing risk toward the development and worsening of PEW as chronic kidney disease progresses. A main cause of PEW is inflammation, which may occur with or without clinically evident illness and can be associated with the most severe forms of PEW. Another major cause of PEW is decreased nutrient intake relative to the patient's nutritional needs, and may be caused by anorexia, which may be engendered by uremic toxicity, emotional depression, medications, or inflammatory disorders. Nonanorexic causes of reduced nutrient intake include inadequate finances to purchase or prepare foods; medical or surgical illnesses that impair the person's ability to ingest, digest, assimilate, or process the nutrients; impaired cognitive function; other mental or physical disabilities; and loss of dentures. Losses of nutrients during dialysis treatments or in urine (eg, the nephrotic syndrome), acidemia, and hormonal disorders can contribute to the development of PEW. Early initiation and adequate doses of renal replacement therapy, rapid treatment of reversible inflammatory processes, ensuring an adequate nutrient intake, and prevention of acidemia may be used to prevent and treat PEW. PMID- 19121474 TI - Potential role of growth factors with particular focus on growth hormone and insulin-like growth factor-1 in the management of chronic kidney disease. AB - Prevention and treatment of chronic kidney disease (CKD) so far primarily has been based on early and aggressive treatment of hypertension. A number of other therapeutic approaches have the potential of being translated to the clinical area within the foreseeable future. In this review, we focus on growth factors and, in particular, on the role of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in the treatment of CKD and the management of its complications. Disturbances of the GH-IGF-1 axis in CKD have suggested therapeutic roles for both the inhibition, as well as the stimulation, of that axis in CKD. Experimental data have suggested that GH may have a detrimental effect on kidney growth, scarring, and the progression of CKD. Therapies based on the inhibition of GH showed some benefit, but failed to translate to clinical CKD. On the other hand, the administration of IGF-1 has been reported in a number of small studies to have conflicting effects on renal function. In end-stage renal disease, GH has been shown in a number of small-scale studies to modify inflammation, improve lipid profile, and favorably affect cardiovascular parameters. Studies also have shown some benefit of IGF-1 administration on the nutritional parameters of patients on renal replacement therapy. More recently, a larger randomized, double blind, placebo-controlled, 26-week, proof-of-concept clinical study was conducted to investigate the effect of GH (Norditropin; Novo Nordisk, Bagsvaerd, Denmark) in adult chronic hemodialysis (CHD) patients. Beneficial effects were observed on lean body mass, serum albumin, high-density lipoprotein cholesterol, and homocysteine levels, all known to be associated with mortality and morbidity in CHD. These studies suggested that GH therapy may prove beneficial in reducing morbidity and mortality in CHD patients. PMID- 19121475 TI - Assessment of body protein: energy status in chronic kidney disease. AB - The prevalence of protein-energy malnutrition progressively increases during the evolution of chronic kidney disease (CKD). As a consequence, it has been reported that 40% of patients present with symptoms of undernutrition at the entrance to chronic dialysis treatment. In patients established on maintenance hemodialysis, the prevalence of malnutrition varies from 20% to 60% according to which indicators of nutritional status are used. Protein-energy malnutrition is associated with an increase in overall and cardiovascular death risks both in CKD patients not yet on dialysis and in dialysis patients. Given the impact of protein-energy wasting on the outcome of CKD patients, screening malnutrition and monitoring protein-energy status appear of primary importance. Therefore, scientific and professional societies or foundations have developed guidelines for the assessment of nutritional status as well as for the treatment of malnourished CKD patients. Recently, an expert panel recommended the term protein energy wasting for loss of body protein mass and fuel reserves. According to these recommendations, protein-energy wasting should be diagnosed if 3 characteristics are present (low serum levels of albumin, transthyretin, or cholesterol), reduced body mass (low or reduced body mass or fat mass or weight loss with reduced intake of protein and energy), and reduced muscle mass (muscle wasting or sarcopenia, reduced mid-arm-muscle circumference). The present article addresses the methods for assessing protein-energy status, their specificities regarding the CKD staging, and the criteria for choosing among these methods when managing the follow-up evaluation of CKD patients. The practical implications of nutritional parameters for the management of CKD patients are illustrated by a case presentation. PMID- 19121476 TI - Correction of metabolic acidosis to ameliorate wasting in chronic kidney disease: goals and strategies. AB - Metabolic acidosis is an important cause of protein-energy wasting, commonly observed in chronic kidney disease (CKD). This wasting is, in part, a result of the imbalance between protein degradation and synthesis induced by metabolic acidosis. The increase in protein degradation seen with metabolic acidosis is largely secondary to increased activities of the adenosine triphosphate dependent, ubiquitin-proteasome system and branched-chain ketoacid dehydrogenase. Studies consistently have shown increased protein degradation with lower serum bicarbonate levels and/or arterial pH; however, the evidence for the anti anabolic effects of metabolic acidosis is less consistent. In contrast to these metabolic studies, many cross-sectional studies have shown a direct relationship between the severity of metabolic acidosis and the adequacy of nutritional status in CKD patients. Moreover, lower serum bicarbonate levels have been associated with better survival in some epidemiologic studies of patients undergoing maintenance hemodialysis. It is likely that these relationships are confounded by the direct association of dietary protein intakes with metabolic acidosis controlling the survival data for measures of dietary protein intakes, malnutrition, and inflammation shows a rather steep increase in the risk of death with lower serum bicarbonate levels. Two randomized controlled studies have shown that correction of metabolic acidosis is associated with reduction in risk for hospitalization in chronic peritoneal dialysis patients; the studies in maintenance hemodialysis patients have been small and inconsistent. For now, metabolic studies and data from clinical trials lend support to the recommendations made by the Nutrition Workgroup of the Kidney Disease Outcomes Quality Initiative to maintain serum bicarbonate levels of 22 mEq/L or greater in all CKD patients. Limited data suggest that a higher serum bicarbonate level (around 24 mEq/L) may be even more beneficial, particularly in chronic peritoneal dialysis patients. PMID- 19121477 TI - Nutrition support for the chronically wasted or acutely catabolic chronic kidney disease patient. AB - Because of the number of factors affecting the nutritional and metabolic status in patients with advanced chronic kidney disease or who are on maintenance dialysis, the prevention and treatment of protein-energy wasting (PEW) of chronic kidney disease should involve a comprehensive combination of maneuvers to diminish protein and energy depletion, in addition to therapies that will avoid further losses. The available evidence suggests that nutritional supplementation, administered orally or parenterally, is effective in the treatment of maintenance dialysis patients with PEW in whom oral dietary intake from regular meals cannot maintain adequate nutritional status. Increased oral nutrient intake during dialysis and at home is the ideal choice for this intervention. In clinical practice, the advantages of intradialytic oral nutritional supplements include proven efficacy and compliance. Therefore, at a minimum, oral nutritional supplementation given intradialytically should be attempted in maintenance dialysis patients with PEW, accompanied by individualized dietary advice for appropriate intake at home. In ones who cannot tolerate oral feeding, other forms of nutritional supplementation including intradialytic parenteral nutritional are a reasonable strategy. Although not proven conclusively, nutritional interventions in the form of supplementation may lead to considerable improvements in mortality, hospitalization, and treatment costs. PMID- 19121478 TI - Vitamin D therapy for chronic kidney disease. AB - Vitamin D has played a central role in the nephrologist's armamentarium, with active vitamin D analogues enjoying broad use for treatment of secondary hyperparathyroidism. Increasing data are now coming to light about the broader biological actions of vitamin D, including wide-ranging effects in several endocrine pathways, cardiovascular disease, infectious disease, and even the progression of chronic kidney disease (CKD). As additional agents are emerging to help with control of metabolic bone disease, these nontraditional pathways of vitamin D action will become increasingly important to consider when formulating a treatment plan. Although the only approved use for vitamin D analogues in CKD is the treatment of secondary hyperparathyroidism, well-conducted clinical trials may soon broaden the scope of this therapy. This article reviews the role of vitamin D therapy in CKD and looks to the answers that future research may bring. PMID- 19121482 TI - The curve of Spee. PMID- 19121483 TI - Divine proportions in the growing face. PMID- 19121484 TI - Is your practice turning green? PMID- 19121486 TI - Orthodontic bracket lost in airway. PMID- 19121487 TI - Treatment effects of mini-implants for en-masse retraction of anterior teeth. PMID- 19121489 TI - Orthodontic appliances and MRIs. PMID- 19121491 TI - Characterization of retrieved orthodontic miniscrew implants. AB - INTRODUCTION: The purposes of this study were to characterize the morphologic, structural, and compositional alterations and to assess any hardness changes in used orthodontic miniscrew implants. METHODS: Eleven miniscrew implants (Aarhus Anchorage System, Medicon eG, Tuttlingen, Germany) placed in 5 patients were retrieved after successful service of 3.5 to 17.5 months; none showed signs of mobility or failure. These implants, and brand-, type-, and size-matched specimens as controls, were subjected to multi-technique characterization. RESULTS: Optical microscopy indicated loss of gloss with variable discoloration. Scanning electron microscopy and x-ray microanalysis showed morphologic alteration of the miniscrew implant surfaces with integuments formed on the surface. The materials precipitated on the surfaces were sodium, potassium, chlorine, iron, calcium, and phosphorus from the contact of the implant with biologic fluids such as blood and exudates, forming sodium chloride, potassium chloride, and calcium-phosphorus precipitates. The composition of the implant was similar to that of a titanium alloy. X-ray microtomography analysis showed no bulk structure alterations. Vickers microhardness testing showed no increased bulk or surface hardness of the retrieved specimens compared with the controls, excluding the possibility of strain-hardening phenomena as a result of self tapping and self-drilling placement and related loading conditions. CONCLUSIONS: Used titanium-alloy miniscrew implants have morphologic and surface structural alterations including adsorption of an integument that is calcified as a result of contact of the implants with biologic fluids. Randomly organized osseointegration islets on these smooth titanium-alloy miniscrew surfaces might be enhanced by the extended period of retention in alveolar bone in spite of the smooth surface and immediate loading pattern of these implants. PMID- 19121493 TI - Effect of bracket base conditioning. AB - INTRODUCTION: The aim of this study was to compare the effect of a silicoating system, the influence of sandblasting, and the effect of a silane-coupling agent after sandblasting on the shear bond strength of stainless steel foil-mesh brackets. To simulate the oral environment, all specimens were thermocycled (6000 times at 5 degrees C and 55 degrees C) in a mastication device before testing. METHODS: Four bracket groups were tested: group 1 consisted of 20 metal brackets that were sandblasted on the base; group 2 contained 20 brackets that were sandblasted, and a silane-coupling agent was applied; in group 3, the surface of the base of 20 metal brackets was treated by using a tribochemical system; and group 4 was the control group. The brackets were bonded with a light-curing adhesive to extracted third molars, and the shear bond strength and the adhesive remnant index score were determined. The brackets of group 1 were reconditioned after debonding with sandblasting and tested again (group 5). RESULTS: Sandblasting and tribochemical treatment of brackets improved the shear bond strength of stainless steel brackets. Combined sandblasting and silane-coupling treatment offers no benefit of increased in-vitro strength. CONCLUSIONS: The bond of resins to tribochemically silicoated stainless steel brackets seems to be sufficient to strengthen the bond between the adhesive and the metal bracket. This treatment is mainly indicated for low-compliance patients or teeth that are difficult to bond. PMID- 19121495 TI - Effect of fluoride-releasing light-cured resin on shear bond strength of orthodontic brackets. AB - INTRODUCTION: The aim of this study was to analyze the effect of an enamel protective resin on the shear bond strength (SBS) of orthodontic brackets bonded with self-etching primer. METHODS: Eighty extracted premolars were randomly divided into 4 groups of 20, and metal brackets were bonded. Group 1 specimens were bonded with Transbond Plus self-etching primer (3M Unitek, Monrovia, Calif) and no fluoride resin; in group 2, a fluoride-releasing resin (Ortho-Coat, Pulpdent, Watertown, Mass) was used with the Transbond Plus self-etching primer; group 3 teeth were bonded with a new antimicrobial self-etching primer (Clearfil Protect Bond, Kuraray Medical, Osaka, Japan) with no fluoride resin; and the same protocol was used in group 4 with an application of Ortho-Coat. A universal testing machine was used to determine the SBS, and the adhesive remaining after debonding was assessed. RESULTS: There was no difference in SBS whether fluoride releasing resin was used. Groups 3 and 4 had higher SBS values than the other groups (P <0.001). The Kruskal-Wallis test showed no significant differences in the adhesive remnant index (chi-square = 0.019, P = 0.990). CONCLUSIONS: The application of enamel-protective resin did not affect the bond strength of orthodontic brackets to enamel with self-etching primer systems. PMID- 19121496 TI - Medication effects on the rate of orthodontic tooth movement: a systematic literature review. AB - INTRODUCTION: Recently, several reviews have been published on the effects of medications on bone physiology and the clinical side effects in orthodontics. However, the effects of medications on the rate of orthodontic tooth movement have not been evaluated. METHODS: A systematic literature review on the effects of medications and dietary supplements on the rate of experimental tooth movement was performed by using PubMed (1953-Oct 2007), Web of Science, and Biosis, complemented by a hand search. RESULTS: Forty-nine articles were included in the review, but their interpretation was hindered by the variability in experimental design, magnitude of force applied during tooth movement, and medication regimens. Therapeutic administration of eicosanoids resulted in increased tooth movement, whereas their blocking led to a decrease. Nonsteroidal anti inflammatory drugs (NSAIDs) decreased tooth movement, but non-NSAID analgesics, such as paracetamol (acetaminophen), had no effect. Corticosteroid hormones, parathyroid hormone, and thyroxin have all been shown to increase tooth movement. Estrogens probably reduce tooth movement, although no direct evidence is available. Vitamin D3 stimulates tooth movement, and dietary calcium seemed to reduce it. Bisphosphonates had a strong inhibitory effect. CONCLUSIONS: Medications might have an important influence on the rate of tooth movement, and information on their consumption is essential to adequately discuss treatment planning with patients. PMID- 19121497 TI - How well does Invisalign work? A prospective clinical study evaluating the efficacy of tooth movement with Invisalign. AB - INTRODUCTION: The purpose of this prospective clinical study was to evaluate the efficacy of tooth movement with removable polyurethane aligners (Invisalign, Align Technology, Santa Clara, Calif). METHODS: The study sample included 37 patients treated with Anterior Invisalign. Four hundred one anterior teeth (198 maxillary and 203 mandibular) were measured on the virtual Treat models. The virtual model of the predicted tooth position was superimposed over the virtual model of the achieved tooth position, created from the posttreatment impression, and the 2 models were superimposed over their stationary posterior teeth by using ToothMeasure, Invisalign's proprietary superimposition software. The amount of tooth movement predicted was compared with the amount achieved after treatment. The types of movements studied were expansion, constriction, intrusion, extrusion, mesiodistal tip, labiolingual tip, and rotation. RESULTS: The mean accuracy of tooth movement with Invisalign was 41%. The most accurate movement was lingual constriction (47.1%), and the least accurate movement was extrusion (29.6%)- specifically, extrusion of the maxillary (18.3%) and mandibular (24.5%) central incisors, followed by mesiodistal tipping of the mandibular canines (26.9%). The accuracy of canine rotation was significantly lower than that of all other teeth, with the exception of the maxillary lateral incisors. At rotational movements greater than 15 degrees, the accuracy of rotation for the maxillary canines fell significantly. Lingual crown tip was significantly more accurate than labial crown tip, particularly for the maxillary incisors. There was no statistical difference in accuracy between maxillary and mandibular teeth of the same tooth type for any movements studied. CONCLUSIONS: We still have much to learn regarding the biomechanics and efficacy of the Invisalign system. A better understanding of Invisalign's ability to move teeth might help the clinician select suitable patients for treatment, guide the proper sequencing of movement, and reduce the need for case refinement. PMID- 19121498 TI - Experimental determination of optimal force system required for control of anterior tooth movement in sliding mechanics. AB - INTRODUCTION: This study was designed to determine the optimum vertical height of the retraction force on the power arm that is required for efficient anterior tooth retraction during space closure with sliding mechanics. METHODS: Three adults (1 man, 2 women) with Angle Class II Division 1 malocclusions were selected for this study. In each subject, the maxillary right central incisor was the target tooth. Initial tooth displacements of that tooth with sliding mechanics with various heights of retraction forces were measured in vivo by a 2 point 3-dimensional displacement magnetic sensor device. The tooth's motion trajectories on the midsagittal plane were studied. RESULTS: The location of the center of rotation of the target tooth varied according to the different heights of the retraction forces. Controlled anterior tooth movement (ie, lingual-crown tipping, lingual-root movement) can be predicted, simulated, or even manipulated by different heights of retraction forces on the power arm in the sliding mechanics force system. A power arm length of 3 to 5 mm is estimated to produce controlled lingual-crown tipping (with the apex as the center of rotation) for efficient anterior tooth retraction during sliding space closure in adults with Angle Class II Division 1 malocclusion. CONCLUSIONS: Knowing and applying the correct height of retraction force on the power arm is the key to efficient anterior tooth retraction. PMID- 19121499 TI - Evaluation of esthetic brackets' resistance to torsional forces from the archwire. AB - INTRODUCTION: The aim of this study was to evaluate the resistance to deformation or fracture of esthetic brackets produced by archwire torsion. METHODS: Six types of maxillary right central incisor brackets were analyzed: traditional ceramic brackets (cer); ceramic brackets reinforced with a stainless steel slot (cer/ss); ceramic brackets reinforced with a gold slot (cer/gold); traditional polycarbonate brackets (poly); polycarbonate brackets reinforced with a stainless steel slot (poly/ss); and polycarbonate brackets reinforced with ceramic fillers and a stainless steel slot (poly/cer/ss). Stainless steel wire segments were used, and the testing instrument (Emic DL 10000, Sao Jose do Rio Preto, PR, Brazil) was moved at a rate of 1 inch per minute to generate the wire torsion. RESULTS: The brackets showed deformation or fracture resistance values (gf x mm) in decreasing order as follows: cer/ss (3528.1 +/- 516.6), cer/gold (2858.7 +/- 611.6), cer (2424.0 +/- 352.1), poly/cer/ss (2279.5 +/- 174.5), poly/ss (2142.0 +/- 275.7), and poly (1463.6 +/- 193.3). The cer/ss ceramic brackets showed the greatest statistically significant (P <0.01) values of resistance to fracture, and the poly brackets had the lowest statistically significant (P <0.01) values of resistance to deformation. The cer brackets showed no significant differences (P >0.01) from the cer/gold, the poly/cer/ss, and the poly/ss brackets. CONCLUSIONS: This suggested that the stainless steel slot might enhance resistance to deformation or fracture, although gold slots and ceramic fillers are ineffective for reinforcing esthetic brackets. PMID- 19121500 TI - Bond failure of gingivally offset mandibular premolar brackets: a randomized controlled clinical trial. AB - INTRODUCTION: The purpose of this randomized controlled clinical trial was to examine the clinical bond failure rate of gingivally offset mandibular premolar brackets and compare it with that of standard mandibular premolar brackets. METHODS: Eighty-three patients were enrolled in this trial. A total of 240 brackets--120 standard and 120 offset--were bonded in a split-mouth design. RESULTS: Ten standard brackets and 1 offset bracket debonded during the trial period. Survival times were analyzed by using the Kaplan-Meier nonparametric test, and comparisons between bracket types were made with the Mantel-Haenszel log rank test. These tests showed that the failure rate between the bracket types was statistically significant (P <0.0058). CONCLUSIONS: Gingivally offset mandibular premolar brackets have a lower bond failure rate than standard mandibular premolar brackets. PMID- 19121501 TI - Shear modulus of 5 flowable composites to the EverStick Ortho fiber-reinforced composite retainer: an in-vitro study. AB - INTRODUCTION: Although fiber-reinforced composites (FRC) are now available for use as orthodontic retainers, little is known about their bonding properties. Our aim in this study was to investigate the adhesive properties of various composites to a commercially available FRC retainer. METHODS: Five flowable composites (Grandio Flow [Voco GmbH, Cuxhaven, Germany], Synergy Flow [Coltene/Whaledent AG, Genf, Switzerland], Tetric Flow [Ivoclar Vivadent AG, Schaan Furstentum, Liechtenstein], Tetric Flow Chroma [Ivoclar Vivadent AG], and Transbond LR [3M Unitek, Monrovia, Calif) were tested for their shear bond strengths to the EverStick Ortho (Stick Tech Ltd Oy, Turku, Finland) FRC retainer stick. Each group contained 15 samples and underwent 1000 aging cycles between 5 degrees C and 55 degrees C. A testing machine was used to measure the shear bond strengths at the composite-FRC interface. All specimens were visually controlled for the location of the fracture line. RESULTS: Typical shear bond strengths were measured at 40 N or 8 MPa. No significant difference was found between the 5 composites (ANOVA, P < or = 0.05). The fracture line was consistently in the FRC retainer. CONCLUSIONS: All composites had similar bonding characteristics, and visual inspection consistently showed fractures in the FRC retainer. Because the point of least resistance was in the retainer, we concluded that all tested composites were equally effective. PMID- 19121502 TI - Nickel concentration in the saliva of patients with nickel-titanium orthodontic appliances. AB - INTRODUCTION: The purpose of this study was to examine whether nickel-titanium (Ni-Ti) archwires cause an increase of nickel concentration in the saliva of 18 orthodontic patients to estimate the possible risk of these archwires in patients who have nickel hypersensitivity. METHODS: Saliva samples were collected before orthodontic treatment, after placement of the bands and brackets, 2 weeks later and before placing the Ni-Ti archwires, immediately after placing the Ni-Ti archwires, 4 weeks after placing the wires, and 8 weeks after placing the wires. RESULTS: By using mass spectrometry, no statistically significant differences were found in the nickel concentrations in the samples taken without appliances, in those obtained 2 weeks after placement of the bands and brackets, and 4 and 8 weeks after placement of the archwires. Samples taken immediately after placement of the bands and brackets and the Ni-Ti archwires showed slight but significant increases in nickel concentration of 78 and 56 microg per liter, respectively, compared with the pretreatment value of 34 microg per liter. CONCLUSIONS: Nickel leaching occurred after placement of the bands and brackets and after placement of the Ni-Ti archwires, associated with an increase of the nickel ion concentration in the patient's saliva. This effect decreased within 10 weeks. PMID- 19121503 TI - Profile changes in orthodontic patients treated with mandibular advancement surgery. AB - INTRODUCTION: The potential to improve facial esthetics is often the deciding factor in treatment planning of borderline orthodontic patients who can be treated with either orthognathic surgery or dental camouflage. The purpose of this study was to determine the degree of skeletal and soft-tissue Class II disharmony necessary before a significant esthetic benefit is derived from mandibular advancement surgery. METHODS: Twenty laypeople, 20 orthodontists, and 20 oral surgeons rated the attractiveness of before and after treatment profiles of 20 mandibular advancement patients using a 5-point Likert scale. The Spearman rank correlation tested for relationships between amount of profile change and varying pretreatment ANB and profile angles. Plots of the distribution of profile changes with varying ANB and profile angles were then examined. RESULTS: There was a tendency for inverse correlations between profile change and profile angle, and for positive correlations between profile change and ANB angles, but only the relationship between profile change and ANB angles judged by the orthodontists was statistically significant (P <0.05). Orthodontists, oral surgeons, and laypeople found that profiles consistently improved when profile angles were < or = 159 degrees, < or = 158 degrees, and < or = 157 degrees, respectively. Orthodontists and oral surgeons found profiles consistently improved when ANB angles were > or = 5.5 degrees and > or = 6.5 degrees, respectively, whereas laypeople showed no trend between ANB angle and profile change. The incidence of having less desirable profiles after treatment was 2.6 to 5.0 times higher when the pretreatment profile angles were larger than the threshold profile angles, and 4.5 to 7.9 times higher when the pretreatment ANB angles were less than threshold ANB angles. CONCLUSIONS: Pretreatment profile angles < 160 degrees and ANB angles > 6 degrees are necessary for profiles to be consistently perceived as improved after surgery and to minimize the incidence of the profile worsening after treatment. PMID- 19121504 TI - Influence of anteroposterior mandibular positions on facial attractiveness in Japanese adults. AB - INTRODUCTION: Our aims in this study were to determine the anteroposterior facial relationship that is regarded as most attractive by Japanese laypersons in a questionnaire survey and to evaluate which analysis of the soft-tissue profile is most suitable for Japanese people. METHODS: We showed 262 Japanese laypersons (121 male, 141 female) 9 morphed profile images with Point B and menton anteriorly or distally moved by software and asked them to number them sequentially according to their attractiveness. To examine which analysis best reflects facial attractiveness as judged by laypersons, we made 5 types of analyses of the facial profile with 11 variables in the 9 images. RESULTS: The normal face was judged favorably; however, an attractive profile might be different for each subject. The 3 highest ranking profiles (normal face and moderate mandibular retrusions) were often favorites, and 2 profiles (severe mandibular protrusions) were liked the least for most subjects. However, the other images showed a wide range of distribution. Mandibular retrusion was generally more favored than mandibular protrusion and bimaxillary protrusion (severe chin retrusion) had a high attractiveness ranking and was well accepted in the Japanese population. CONCLUSIONS: To evaluate the profiles of Japanese subjects, it is important to evaluate not only the esthetic line defined by the nose and chin, but also the balance of the upper and lower lips defined by the posterior reference line--ie, Burstone's Sn-Pog' line. PMID- 19121505 TI - Age-dependant cephalometric standards as determined by multilevel modeling. AB - INTRODUCTION: The purpose of this study was to evaluate the feasibility of constructing age-dependant cephalometric standards for white subjects by using 3 data sets. METHODS: The data sets were the samples from the Fels Longitudinal Study (United States), the Michigan Growth Study (United States), and the Nijmegen Growth Study (The Netherlands). The 3 mixed-longitudinal samples provided data for 218 girls and 231 boys between 9 and 14 years of age and were compared based on 4 cephalometric angles: SNA, SNB, ANB, and SN/GoMe. Curve fitting and statistical comparisons were performed with multilevel modeling procedures. RESULTS: All 4 angles showed linear changes over time. SNA and SNB increased, whereas ANB and SN/GoMe decreased. Based on paired-sample comparisons, the samples displayed statistically significant (P <0.05) differences for 50% of the growth velocities and 8% to 17% of the intercepts (size of the angle at 11 years). The SNA and SNB angles showed small and inconsistent differences across the samples. The ANB angle for the Fels boys decreased less than in the other 2 samples. The Nijmegen and Fels girls had the greatest and the least decreases, respectively, in the SN/GoMe angle. Most sample differences decreased over time. CONCLUSIONS: Based on the growth differences identified, we concluded that sagittal and vertical jaw relationships have different patterns of growth in different samples of white subjects; indiscriminate pooling of data, to create age-dependant cephalometric standards for white subjects is not recommended. PMID- 19121506 TI - Computerized axiographic evaluation of the changes in sagittal condylar path inclination with dental and physical development. AB - INTRODUCTION: The aim of this study was to quantify the changes in sagittal condylar path inclination during maximum mandibular protrusion in relation to the stage of dental and physical development throughout the mixed dentition period. METHODS: A total of 161 children, 6.5 to 13 years of age, were divided by dental stage into 3 subgroups: early mixed dentition without anterior guidance, early mixed dentition with anterior guidance, and late mixed dentition. The control group consisted of 41 adults (mean age, 28 years). All subjects had normal temporomandibular joint function and neutral occlusion. Five maximum protrusion retrusion movements were recorded by using an ultrasound jaw-tracking system supported with software that automatically calculated the condylar path inclination angle (CPIA). RESULTS: Significant differences were found between the 3 subgroups of children. In the late mixed dentition subgroup, girls had significantly steeper condylar paths on the left side than boys. In the early mixed dentition with anterior guidance subgroup, the right CPIA was significantly larger than the left CPIA. Linear regression analysis showed that condylar path inclination was positively correlated with both body height and weight, although the correlation was statistically weak. CONCLUSIONS: These findings indicate that CPIA increases with the maturation of occlusion. PMID- 19121507 TI - Periodontal pathogen levels in adolescents before, during, and after fixed orthodontic appliance therapy. AB - INTRODUCTION: This purpose of this study was to document and investigate changes in periodontal pathogen levels before, during, and after orthodontic treatment in adolescents. METHODS: DNA gene probe analysis was used to quantify the levels of 8 periodontal pathogens before, during, and after treatment with fixed orthodontic appliances in 190 concurrently treated adolescent orthodontic patients. The 8 pathogens examined were Actinobacillus actinomycetemcomitans (AA), Porphyromonas gingivalis (PG), Prevotella intermedia (PI), Tannerella forsythia (TF), Eikenella corrodens (EC), Fusobacterium nucleatum (FN), Treponema denticola (TD), and Campylobacter rectus (CR). Chi-square tests were used to determine whether the percentages of subjects with high counts significantly changed over time. Logistic regression analyses were also performed to derive the relative risk of higher counts of pathogenic bacteria with fixed appliances at the various time intervals studied. RESULTS: For 6 (PI, TF, EC, FN, TD, CR) of the 8 pathogens, the percentages of subjects with high pathogen counts increased significantly after 6 months of fixed appliance treatment, but these returned to pretreatment levels by 12 months of orthodontic treatment. No pathogen level was significantly higher after 12 months of orthodontic treatment, and orthodontic treatment was found to be significantly protective for half of the pathogens (EC, FN, TD, CR) posttreatment. CONCLUSIONS: Orthodontic treatment with fixed appliances does not increase the risk of high levels of these periodontal pathogens. PMID- 19121508 TI - Cystatins and cathepsin B during orthodontic tooth movement. AB - INTRODUCTION: The lysosomal cysteine protease cathepsin B is known to play an important role in the resolution of organic matrix, a final step in bone resorption. Cystatins function as an inhibitor of cathepsin B. Determining the correlation between cathepsin B and cystatin levels in gingival crevicular fluid at various times might provide a better understanding of both the dynamics and the metabolic stages of orthodontic tooth movement. METHODS: Human gingival crevicular fluid was collected at the distal sulcus from the canines of persons not in orthodontic treatment, in retention, and in retraction at various times (initial, 1 day, 1 week, and 1 month postretraction). Cathepsin B and its inhibitor, cystatin, were found with fluorometry. RESULTS: The level of cathepsin B was varied in the retraction group; this was different from the retention and the nonorthodontic groups. Significant initial decreases after force application and subsequent increases by 1 month posttreatment were observed in the retraction group. The variations and differences among groups were negatively correlated with cystatin. CONCLUSIONS: The balance between enzyme and inhibitor might reflect the clinical status of orthodontic tooth movement and provide valuable information for the assessment of recall intervals and retention procedures. PMID- 19121509 TI - Effect of lip bumpers on mandibular arch dimensions. AB - INTRODUCTION: The aim of this systematic review was to examine the effects of lip bumper therapy on mandibular arch dimensions. METHODS: A literature survey of PubMed, EMBASE, Cochrane Central, and Cochrane Database of Systematic Reviews (www.cochrane.org) was conducted from December 1968 to January 2007. Human studies, randomized clinical trials, prospective and retrospective studies, and studies discussing the effect of lip bumpers on the arch and teeth were included. Two reviewers independently selected and extracted the data. RESULTS: Of the 52 studies found in the search, only 1 met the inclusion criteria. CONCLUSIONS: The results showed increases in arch dimensions that included an increase in arch length. This was attributed to incisor proclination, distalization, and distal tipping of the molars. There was also an increase in the arch width seen in the intercanine and deciduous intermolar and premolar distances. The long-term stability of the effects of the lip bumper need to be elucidated. PMID- 19121510 TI - Two-component mini-implant as an efficient tool for orthognathic patients. AB - A new type of orthodontic mini-implant called the C-implant can be an effective alternative to conventional 1-component screws in orthognathic patients because of its particular design. Its small size and 2-component structure make it easily applicable for various types of difficult intermaxillary fixation cases such as 2 jaw orthognathic surgery, 1-jaw surgery with genioplasty, and orthognathic surgery without presurgical orthodontic treatment. The 2-part design highly resists fracture or deformation during placement and removal, and the long-span head allows the patient to easily attach intermaxillary elastics for traction. This mini-implant can be used not only as an intermaxillary fixation screw but also as anchorage during presurgical and postsurgical orthodontic treatment. Better osseointegration potential of the surface-treated screw part allows it to endure heavy and dynamic forces. In this article, we attempted to show that this mini-implant is a good tool for effective anchorage in presurgical treatment, intermaxillary fixation during surgery, and postsurgical treatment of orthognathic patients. PMID- 19121512 TI - Category 2: Adult malocclusion with open bite. AB - This case report was displayed by the American Board of Orthodontics at the Annual Session of the American Association of Orthodontists, as an example of a successfully completed case from the 2006 ABO clinical exam. PMID- 19121511 TI - Nonsurgical treatment of an adult with mandibular asymmetry and unilateral posterior crossbite. AB - The patient was a woman, 24 years 8 months old, with mandibular asymmetry resulting in a unilateral posterior crossbite. In addition to moderate crowding in the mandibular arch, the malocclusion was further complicated by an asymmetric maxillary arch, a consequence of an extracted maxillary left second premolar. The mandibular arch was skewed, and there were severe dental compensations. The patient was treated nonsurgically, with unilateral extractions of teeth and asymmetric mechanics. Pretreatment and posttreatment records are shown, and the treatment strategy resulting in few side effects is described. PMID- 19121513 TI - Litigation, legislation, and ethics. When two heads are not better than one. PMID- 19121514 TI - Introducing a new entity into discourse: comprehension and production evidence for the status of Dutch er "there" as a higher-level expectancy monitor. AB - This paper reports on the ways in which new entities are introduced into discourse. First, we present the evidence in support of a model of indefinite reference processing based on three principles: the listener's ability to make predictive inferences in order to decrease the unexpectedness of upcoming words, the availability to the speaker of grammatical constructions that customize predictive inferences, and the use of "expectancy monitors" to signal and facilitate the introduction of highly unpredictable entities. We provide evidence that one of these expectancy monitors in Dutch is the post-verbal variant of existential er (the equivalent of the unstressed existential "there" in English). In an eye-tracking experiment we demonstrate that the presence of er decreases the processing difficulties caused by low subject expectancy. A corpus-based regression analysis subsequently confirms that the production of er is determined almost exclusively by seven parameters of low subject expectancy. Together, the comprehension and production data suggest that while existential er functions as an expectancy monitor in much the same way as speech disfluencies (hesitations, pauses and filled pauses), er is a higher-level expectancy monitor because it is available in spoken and written discourse and because it is produced more systematically than any disfluency. PMID- 19121515 TI - Effects of magnification methods and magnifier shapes on visual inspection. AB - This study investigated the effectiveness of the novel differential linear and differential nonlinear magnification methods, and the effects of magnifier shape and magnification power on visual search performance. With the differential magnification, objects that were more peripheral were scaled at a level higher than those in the vicinity of fixation. It was hypothesized that 1) owing to the ineffectiveness of scaling stimulus in the vicinity of fixation, the differential magnification methods would enhance magnification effectiveness and thereby improve inspection performance, and 2) because an elliptical lens shape resembles the shape of human visual field, for the same area of magnified view, an elliptical magnifier would be more effective than the circular one. In this study, objective performance measures of speed and accuracy and subjective workload evaluation using the NASA Task Load Index paradigm were used. The results showed that 4x magnification increased the overall inspection time as compared with 3x magnification, suggesting that a trade-off might have occurred between magnification and field size. The two hypotheses were not confirmed; however, a number of questions arose which need further investigation. Discussions of the concerned questions were made on i) the unfamiliarity of subjects with the new differential magnification methods, ii) perceptual discomfort due to image duplication, discontinuity, and distortion resulting from stimulus scaling, iii) the movement of fixation points, and iv) the competition for attention between foveal and peripheral magnified objects. This study prompts the authors to consider a training programme on the differential linear and nonlinear magnification methods in future studies. With proper instructions regarding the nature of lens and magnified images, subjects' performance with the novel differential magnification methods may be improved. PMID- 19121516 TI - Delay discounting correlates with proportional lateral frontal cortex volumes. AB - BACKGROUND: Functional neuroimaging experiments in healthy control subjects have shown that choosing between small and immediate rewards versus larger but deferred rewards in delay discounting (DD) tasks recruits mesofrontal and lateral frontal cortex. Might individual differences in frontocortical gray matter morphology be related to preference for immediate reward? METHODS: We related DD in a laboratory decision-making task to proportional frontocortical gray matter (GM) volumes calculated from segmented magnetic resonance images in 29 healthy adults. RESULTS: Dorsolateral and inferolateral frontal cortex GM volumes (corrected as a proportion of whole cerebral brain volume) each correlated inversely with preference for immediate gratification during decision making, as indexed by DD constant k. Conversely, neither proportional orbitofrontal or mesofrontal cortex GM volume nor cerebral brain volume (CBV) or total intracranial volume (ICV; a measure of maximal brain growth) significantly correlated with severity of DD. CONCLUSIONS: Severity of discounting of delayed rewards correlates with proportional lateral frontocortical GM morphology but not with whole brain measures. In light of evidence of frontocortical abnormalities in substance dependence and sociopathy, future studies can assess whether reduced frontocortical volume itself is a morphological marker or risk factor for inability to delay gratification in psychiatric disorders. PMID- 19121517 TI - Molecular determinants of dysregulated GABAergic gene expression in the prefrontal cortex of subjects with schizophrenia. AB - BACKGROUND: Prefrontal deficits in gamma-aminobutyric acid (GABA)ergic gene expression, including neuropeptide Y (NPY), somatostatin (SST), and parvalbumin (PV) messenger RNAs (mRNAs), have been reported for multiple schizophrenia cohorts. Preclinical models suggest that a subset of these GABAergic markers (NPY/SST) is regulated by brain-derived neurotrophic factor (BDNF), which in turn is under the inhibitory influence of small noncoding RNAs. However, it remains unclear if these mechanisms are important determinants for dysregulated NPY and SST expression in prefrontal cortex (PFC) of subjects with schizophrenia. METHODS: Using a postmortem case-control design, the association between BDNF protein, NPY/SST/PV mRNAs, and two BDNF-regulating microRNAs (miR-195 and miR-30a 5p) was determined in samples from the PFC of 20 schizophrenia and 20 control subjects. Complementary studies were conducted in cerebral cortex of mice subjected to antipsychotic treatment or a brain-specific ablation of the Bdnf gene. RESULTS: Subjects with schizophrenia showed deficits in NPY and PV mRNAs. Within-pair differences in BDNF protein levels showed strong positive correlations with NPY and SST and a robust inverse association with miR-195 levels, which in turn were not affected by antipsychotic treatment or genetic ablation of Bdnf. CONCLUSIONS: Taken together, these results suggest that prefrontal deficits in a subset of GABAergic mRNAs, including NPY, are dependent on the regional supply of BDNF, which in turn is fine-tuned through a microRNA (miRNA)-mediated mechanism. PMID- 19121518 TI - Flexible shaping: how learning in small steps helps. AB - Humans and animals can perform much more complex tasks than they can acquire using pure trial and error learning. This gap is filled by teaching. One important method of instruction is shaping, in which a teacher decomposes a complete task into sub-components, thereby providing an easier path to learning. Despite its importance, shaping has not been substantially studied in the context of computational modeling of cognitive learning. Here we study the shaping of a hierarchical working memory task using an abstract neural network model as the target learner. Shaping significantly boosts the speed of acquisition of the task compared with conventional training, to a degree that increases with the temporal complexity of the task. Further, it leads to internal representations that are more robust to task manipulations such as reversals. We use the model to investigate some of the elements of successful shaping. PMID- 19121519 TI - When knowing can replace seeing in audiovisual integration of actions. AB - The ability to predict the effects of actions is necessary to behave properly in our physical and social world. Here, we describe how the ability to predict the consequence of complex gestures can change the way we integrate sight and sound when relevant visual information is missing. Six drummers and six novices were asked to judge audiovisual synchrony for drumming point-light displays where the visual information was manipulated to eliminate or include the drumstick-drumhead impact point. In the condition with only the arm information novices were unable to detect asynchrony whereas drummers were able to. Additionally, in the conditions that included the impact point drummers perceived the best alignment when the sight preceded the sound, while in the arm only condition they perceived the best alignment when the sound occurred together with or preceded the sight, as it would be expected if they were predicting the beat occurrence. Taken together these findings suggest that humans can acquire, through practice, internal models of action which can be used to replace missing information when integrating multisensory signals from the environment. PMID- 19121520 TI - The evaluation of EEG response to photic stimulation in normal and diseased subjects. AB - In this paper, our aim is to determine two photic stimulation frequencies, which would represent normal and diseased subjects, separately. Following features were extracted for this aim; linear prediction coefficients (LPC), subband wavelet entropy (SWE), subband wavelet variance (SWV), and relative power (RP). After extracting related features, analysis of variance (ANOVA) statistical test was used for the statistical evaluation of these features. According to the obtained results, wavelet transform-based entropy gave the best results to determine the representing stimulation frequencies. As a result, 29 Hz stimulation frequency was determined as the most representative frequency for normal subjects, whereas 8 Hz stimulation frequency was determined as the most representative frequency for diseased subjects. PMID- 19121521 TI - Foreign accent syndrome as a developmental motor speech disorder. AB - INTRODUCTION: Foreign Accent Syndrome (FAS) is a relatively rare motor speech disorder in which the pronunciation of a patient is perceived by listeners of the same language community as distinctly foreign. FAS has been well documented in adult patients with etiologically heterogeneous, though mostly vascular brain lesions affecting the motor speech network of the language dominant hemisphere. In addition, reports exist of adult patients in whom FAS was due to a psychiatric illness. Although FAS has been reported in children, such accounts are rare and have remained largely anecdotal in that there have been no formally documented cases of FAS as a developmental motor speech disorder. METHODS AND RESULTS: For the first time, we describe the clinical, cognitive and neurolinguistic findings in two patients who in the absence of a history of psychiatric illness or acquired brain damage already presented with FAS at an early stage of speech and language development. In the first patient "developmental FAS" was associated with a dysharmonic distribution of neurocognitive test results indicating slight underdevelopment of visuo-spatial skills and visual memory. The second patient presented with "developmental FAS" associated with specific language impairment (SLI). Independent support for a diagnosis of FAS in both patients was obtained in an accent attribution experiment in which groups of native speakers of (Belgian) Dutch assessed the type of foreign accent of a sample of the patients' conversational speech. Both patients were judged as non-native speakers of Dutch by the majority of participants who predominantly identified the accent as French. CONCLUSION: This paper for the first time documents two patients who presented with FAS on a developmental basis. The finding that FAS does not only occur in the context of acquired brain damage or psychogenic illness but also exists as developmental motor speech impairment requires a re-definition of FAS as a clinical syndrome. PMID- 19121522 TI - Gait analysis using gravitational acceleration measured by wearable sensors. AB - A novel method for measuring human gait posture using wearable sensor units is proposed. The sensor units consist of a tri-axial acceleration sensor and three gyro sensors aligned on three axes. The acceleration and angular velocity during walking were measured with seven sensor units worn on the abdomen and the lower limb segments (both thighs, shanks and feet). The three-dimensional positions of each joint are calculated from each segment length and joint angle. Joint angle can be estimated mechanically from the gravitational acceleration along the anterior axis of the segment. However, the acceleration data during walking includes three major components; translational acceleration, gravitational acceleration and external noise. Therefore, an optimization analysis was represented to separate only the gravitational acceleration from the acceleration data. Because the cyclic patterns of acceleration data can be found during constant walking, a FFT analysis was applied to obtain some characteristic frequencies in it. A pattern of gravitational acceleration was assumed using some parts of these characteristic frequencies. Every joint position was calculated from the pattern under the condition of physiological motion range of each joint. An optimized pattern of the gravitational acceleration was selected as a solution of an inverse problem. Gaits of three healthy volunteers were measured by walking for 20s on a flat floor. As a result, the acceleration data of every segment was measured simultaneously. The characteristic three-dimensional walking could be shown by the expression using a stick figure model. In addition, the trajectories of the knee joint in the horizontal plane could be checked by visual imaging on a PC. Therefore, this method provides important quantitive information for gait diagnosis. PMID- 19121523 TI - Effects of reflex delays on postural control during unstable seated balance. AB - Patients with low-back pain (LBP) exhibit longer trunk muscle reflex latencies and poorer postural control than healthy individuals. We hypothesized that balance during a simulated postural control task would become impaired when the delays exhibited by LBP patients were incorporated into neuromuscular control. The task chosen for this investigation was seated balancing, which emphasizes trunk muscles' contribution in postural control. This task was modeled in Simulink as a fourth order linearized dynamic system with feedback delays. Optimization (minimizing error between experimental and model data) of state variables was used to determine neuromuscular control parameters. Experimental data were obtained from 7 subjects during 5 perturbation trials while balancing on the seat with eyes closed. Model accuracy, reflecting the ability of the model to capture the dynamics of seated balance, was correlated with seated balance performance (r=0.91, p<0.001). To minimize the risk of erroneous findings from inaccurate modeling, only the best five balancers' data were used for hypothesis testing. In these five subjects, feedback delays in modeled neuromuscular control were increased to determine their effect on task stability, trunk displacement and trunk moment. Simulations showed that longer delays found in LBP, in general, did not produce unstable balancing, but did result in increased trunk displacement (p<0.001) and trunk moment (p=0.001). This impairment in neuromuscular control in chronic LBP patients could possibly exacerbate their condition by increasing tissue strain (more spinal displacement) and stress (more spinal loading). PMID- 19121524 TI - Cellular strain assessment tool (CSAT): precision-controlled cyclic uniaxial tensile loading. AB - The development of a multi-sample strain device and elastomeric culture wells designed to systematically assess strain effects on cell cultures is presented in this report. This device enables one to precisely conduct experimental analyses in sterile conditions while delivering cyclic uniaxial tensile strain. The input to the computer interface allows one to alter variables of frequency, duration, and amplitude of strain. The influence of strain on the migration of human umbilical vein endothelial cell (HUVEC) cultured on 2D polydimethylsiloxane (PDMS) surfaces was examined to verify the utility of this system. PMID- 19121525 TI - Prediction of joint center location by customizable multiple regressions: application to clavicle, scapula and humerus. AB - Accurate spatial location of joint center (JC) is a key issue in motion analysis since JC locations are used to define standardized anatomical frames, in which results are represented. Accurate and reproducible JC location is important for data comparison and data exchange. This paper presents a method for JC locations based on the multiple regression algorithms without preliminary assumption on the behavior of the joint-of-interest. Regression equations were obtained from manually palpable ALs on each bone-of-interest. Results are presented for all joint surfaces found on the clavicle, scapula and humeral bone. Mean accuracy errors on the JC locations obtained on dry bones were 5.2+/-2.5 mm for the humeral head, 2.5+/-1.1 mm for the humeral trochlea, 2.3+/-0.9 mm for the humeral capitulum, 8.2+/-3.9 mm for the scapula glenoid cavity, 7.2+/-3.2 mm for the scapular aspect of the acromio-clavicular joint, 3.5+/-1.8mm for the clavicular aspect of the sternoclavicular joint and 3.2+/-1.4 mm for the clavicular aspect of the acromio-clavicular joint. In-vitro and in-vivo validation accuracy was 5.3 and 8.5mm, respectively, for the humeral head center location. Regression coefficients for joint radius dimension and joint surface orientation were also processed and reported in this paper. PMID- 19121526 TI - Unbiased treatment effect estimates by modeling the disease process of multiple sclerosis. AB - Gadolinium-enhancing lesions in the brain are commonly used as a primary outcome measure of disease activity in phase I/II clinical trials in multiple sclerosis (MS). The advent of effective therapy and the cost of clinical trials have led some researchers to adopt a one-arm study design with selection towards patients showing MRI activity. Regression to the mean is recognized as an important consideration in these trials, but the additional confounding effect of alternating active and inactive phases of disease has not been considered. Simulated data were generated from Poisson and normal distributions to mimic outcomes from phase I/II clinical trials of patients with relapsing-remitting MS under a constant or changing disease process model. In all cases, conventional comparison of pretreatment to on-treatment measurements overestimated the treatment effect. Although correction for regression to the mean provided unbiased estimates of the treatment effect under a constant disease process model, this correction also overestimated the treatment effect when disease activity changed over time. Conversely, unbiased estimates of the treatment effect under an alternating (active/inactive) disease process were obtained by correctly accounting for regression to the mean and the disease process. The implications of these results are discussed in terms of efficacy and safety. PMID- 19121527 TI - Total body burden and tissue distribution of polyfluorinated compounds in harbor seals (Phoca vitulina) from the German Bight. AB - Total body burden and tissue distribution of polyfluorinated compounds (PFCs) were investigated in harbor seals (Phoca vitulina) from the German Bight in 2007. A total number of 18 individual PFCs from the following groups could be quantified in the different tissues: perfluorinated carboxylic acids (PFCAs) and perfluorinated sulfonates (PFSAs) and their precursors perfluorinated sulfinates (PFSiAs), perfluorinated sulfonamides, and sulfonamido ethanols. Perfluorooctanesulfonate (PFOS) was the predominant compound in all measured seal tissues (up to 1665 ng g(-1) wet weight in liver tissue). The dominant PFCAs were perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA), but their concentrations were much lower compared to PFOS. The mean whole body burden in harbor seals of all detected PFCs was estimated to be 2665+/-1207 microg absolute. The major amount of the total PFCs burden in the bodies was in blood (38%) and liver (36%), followed by muscle (13%), lung (8%), kidney (2%), blubber (2%), heart (1%), brain (1%), thymus (<0.01%) and thyroid (<0.01%). These data suggest large differences in body burden and accumulation pattern of PFCs in marine mammals. PMID- 19121528 TI - Chromone and chromanone glucosides from Hypericum sikokumontanum and their anti Helicobacter pylori activities. AB - Chromone glucosides, takanechromones A-C (1, 2 and 5) and chromanone glucosides, named takanechromanones A and B (3 and 4), were isolated from the methanolic extracts of Hypericumsikokumontanum together with 27 known compounds. Their structures were established based on spectroscopic evidence. The isolated compounds and some chromone derivatives were assayed for antimicrobial activity against Helicobacter pylori and cytotoxicity against human cancer cell lines. PMID- 19121529 TI - Canine bone marrow cells differentiate into hepatocyte-like cells and placental hydrolysate is a potential inducer. AB - Hepatocyte growth factor (HGF) can stimulate human and rat bone marrow (BM) cells to differentiate into hepatocytes. A human placental hydrolysate (hPH) stimulates proliferation of hepatocytes, but its role as a potential inducer of BM cells to form hepatocytes is unclear. To determine if canine BM cells stimulated with HGF or hPH differentiate into hepatocyte-like cells, BM cells were cultured with HGF or hPH. The cultured cells underwent morphological examination, expression of albumin and cytokeratin 18 (CK18), hepatic function tests including uptake of low density lipoprotein (LDL) and cytochrome P (CYP) 450 activity. Albumin mRNA and protein expression of albumin and CK18 proteins were detected in cultures with HGF and hPH. Furthermore, these cells demonstrated LDL uptake and CYP450 activity. These results indicate that canine BM cells can differentiate into hepatocyte-like cells when stimulated by both HGF and that hPH may be an effective inducer of hepatic differentiation. PMID- 19121530 TI - Habituation of healthy dogs to treadmill trotting: repeatability assessment of vertical ground reaction force. AB - To assess the repeatability of kinetic gait analysis with a treadmill, 28 sound adult dogs were made to trot on an instrumented system. Vertical ground reaction force variables (Peak PFz and Impulse IFz) were collected by 10-s recordings, once a week, 4 weeks in succession. Data were analysed using a repeated-measures two-way ANOVA to investigate habituation to treadmill locomotion. Recorded data were stabilized from the end of the first and second sessions for IFz and PFz, respectively. The percentages of variance attributable to dogs, weeks, minutes and repetitions were, respectively, 72%, 10%, 7%, and 11% for PFz and 84%, 7%, 3%, and 6% for IFz. Habituation occurred after a single training session. Good repeatability was determined by a low coefficient of variation (3.4-4.7%). Adding a treadmill to kinetic gait analysis deserves consideration: reliable data are easily recorded using appropriate habituation and statistical model. PMID- 19121531 TI - Shear horizontal surface acoustic waves in semi-infinite piezoelectrics/metal superlattices. AB - In this paper, an analytical method is presented for the study of shear horizontal surface acoustic waves in semi-infinite piezoelectrics/metal superlattices. The results show that the high electromechanical coupling coefficient and wide usable frequency range can be obtained in these superlattices. PMID- 19121532 TI - Relationship between F-specific RNA phage genogroups, faecal pollution indicators and human adenoviruses in river water. AB - Recent studies have shown the increasing interest of F-specific RNA phage genotyping to identify major sources of faecal contamination in waters. This study, conducted in a river located in an urbanized watershed with recognized anthropogenic influences, was aimed at evaluating the relevance of direct phage genotyping by real-time RT-PCR. One hundred percent of positive results were obtained with a 5 mL aliquot of river water (n=31). Phage distribution was modified after cultivation, since the ratio of the two most abundant genogroups (II and I) reached 1.51 log(10) by direct RT-PCR-based method versus 0.30 log(10) after cultivation (n=8). For the first time, positive correlations between the concentrations of genogroup II, bacterial indicators and human adenoviruses were observed, which may indicate a human faecal pollution. No correlation between genogroups II and I has been revealed. The concentration of genogroup I was only correlated with water turbidity, suggesting an animal pollution coming from upstream after rainfall events. Among the microbiological parameters studied, only genogroup II/genogroup I ratio shows variations occurring in the major sources of faecal pollution. PMID- 19121533 TI - EDTA-assisted Pb phytoextraction. AB - Pb is one of the most widespread and metal pollutants in soil. It is generally concentrated in surface layers with only a minor portion of the total metal found in soil solution. Phytoextraction has been proposed as an inexpensive, sustainable, in situ plant-based technology that makes use of natural hyperaccumulators as well as high biomass producing crops to help rehabilitate soils contaminated with heavy metals without destructive effects on soil properties. The success of phytoextraction is determined by the amount of biomass, concentration of heavy metals in plant, and bioavailable fraction of heavy metals in the rooting medium. In general, metal hyperaccumulators are low biomass, slow growing plant species that are highly metal specific. For some metals such as Pb, there are no hyperaccumulator plant species known to date. Although high biomass-yielding non-hyperaccumulator plants lack an inherent ability to accumulate unusual concentrations of Pb, soil application of chelating agents such as EDTA has been proposed to enhance the metal concentration in above ground harvestable plant parts through enhancing the metal solubility and translocation from roots to shoots. Leaching of metals due to enhanced mobility during EDTA-assisted phytoextraction has been demonstrated as one of the potential hazards associated with this technology. Due to environmental persistence of EDTA in combination with its strong chelating abilities, the scientific community is moving away from the use of EDTA in phytoextraction and is turning to less aggressive alternative strategies such as the use of organic acids or more degradable APCAs (aminopolycarboxylic acids). We have therefore arrived at a point in phytoremediation research history in which we need to distance ourselves from EDTA as a proposed soil amendment within the context of phytoextraction. However, valuable lessons are to be learned from over a decade of EDTA-assisted phytoremediation research when considering the implementation of more degradable alternatives in assisted phytoextraction practices. PMID- 19121534 TI - First Janos Fischer Memorial Symposium. Editorial. PMID- 19121535 TI - Innervation of lung and heart in the ray-finned fish, bichirs. AB - Anatomical and functional studies of the autonomic innervation in the lung and the heart of the bichirs are lacking. The present review paper describes the presence of nerve fibers located in the muscle layers of the lung and its submucosa, the collection of unipolar neurons found in the submucosal and muscle layers of the glottis in a bichir species (Polypterus bichir bichir). Putative oxygen chemoreceptive, neuroepithelial cells (NECs) in the lung mucosa are also included. The latter share many immunohistochemical characteristics similar to those observed in the carotid body and neuroepithelial bodies of mammals. A packed collection of paraganglion cells is located within the trunk of the pulmonary vagus nerves. The paper also examines the occurrence of intracardiac neurons and nerve fibers in the heart of the above species. These studies show that various neurotransmitters may indicate different patterns of innervation in the lung and the heart of the bichirs. However, there is still much to be discovered about the lung and cardiovascular nervous control of these primitive fishes. PMID- 19121536 TI - Topo-optical investigations on the surface of bacterial cells during the phagocytosis of Klebsiella pneumoniae in mouse. AB - Polarisation optical methods provide the means to perform sub-microscopic investigations on structures containing spatially highly ordered molecules, for example the cell envelope of prokaryotic cells. Such structures can evoke birefringence, which can be enhanced or modified by different dyes or reagents, thus providing the possibility of a more specific investigation of the composition and structure of bacterial surface compounds. Klebsiella pneumoniae synthesises sterically different carbohydrate-rich structures, including those of the outermost capsular polysaccharide, the polysaccharide somatic antigen of the lipopolysaccharide molecule and the peptidoglycan layer of the cell wall. In the study reported here, the nature and intensity of topo-optical activity of these structures was analysed using the aldehyde-bisulphite-toluidine blue reaction, sialic acid topo-optical reactions and chlorpromazine-eosin charge transfer reactions. Furthermore, a mouse intraperitoneal model was used to analyse alterations in topo-optical characteristics of bacteria during phagocytosis. Both encapsulated and non-encapsulated bacterial cells changed their original pattern and orientation of birefringence after being phagocytosed. PMID- 19121537 TI - Delivery of sodium borocaptate to glioma cells using immunoliposome conjugated with anti-EGFR antibodies by ZZ-His. AB - Nanoparticles are effective of delivering cargo into cells. Here, sodium borocaptate (BSH) was encapsulated in liposomes composed of nickel lipid, and anti-epidermal growth factor receptor (EGFR) antibodies were conjugated to the liposomes using the antibody affinity motif of protein A (ZZ) as an adaptor (immunoliposomes). The immunoliposomes were used to deliver BSH into EGFR overexpressing glioma cells. Immunohistochemical analysis using an anti-BSH monoclonal antibody revealed that BSH was delivered effectively into the cells but not into EGFR-deficient glioma or primary astrocytes. In an animal model of brain tumors, both the liposomes and the BSH were only observed in the tumor. Moreover, the efficiency of (10)B's delivery into glioma cells was confirmed by inductively coupled plasma-atomic emission spectrometry (ICP-AES) both in vitro and in vivo. The results suggest that this system utilizing immunoliposomes provides an effective means of delivering (10)B into glioma cells in boron neutron capture therapy (BNCT). PMID- 19121539 TI - The use of a shape-memory poly(epsilon-caprolactone)dimethacrylate network as a tissue engineering scaffold. AB - Shape-memory polymers produced from many natural or synthetic raw polymers are able to undergo a shape transformation after exposure to a specific external stimulus. This feature enables their use in minimal-invasive surgery with a small, compact starting material switching over to a more voluminous structure in the body. The use of biomaterials in modern medicine calls for compatibility tests with cell types, encountering the biomaterial during a short-term or long term in vivo application. We analysed the cell behaviour of L929 mouse fibroblasts, human mesenchymal stem cells, human mesothelial cells and rat mesothelial cells on a biodegradable shape-memory polymer network to assess its suitability for medical applications. Further, we investigated the differentiation capacity of mesenchymal stem cells into osteoblasts and adipocytes on the polymer and we analysed the influence of the shape-memory effect on adherent cells. The polymer was cytocompatible for all tested cell types, supporting cell viability and proliferation. The differentiation capacity of mesenchymal stem cells was supported by the polymer and shape-memory effect activation did not affect the majority of adherent cells. PMID- 19121538 TI - Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. AB - Recently, macrophages have been characterized as having an M1 or M2 phenotype based on receptor expression, cytokine and effector molecule production, and function. The effects of macrophage phenotype upon tissue remodeling following the implantation of a biomaterial are largely unknown. The objectives of this study were to determine the effects of a cellular component within an implanted extracellular matrix (ECM) scaffold upon macrophage phenotype, and to determine the relationship between macrophage phenotype and tissue remodeling. Partial thickness defects in the abdominal wall musculature of Sprague-Dawley rats were repaired with autologous body wall tissue, acellular allogeneic rat body wall ECM, xenogeneic pig urinary bladder tissue, or acellular xenogeneic pig urinary bladder ECM. At 3, 7, 14, and 28 days the host tissue response was characterized using histologic, immunohistochemical, and RT-PCR methods. The acellular test articles were shown to elicit a predominantly M2 type response and resulted in constructive remodeling, while those containing a cellular component, even an autologous cellular component, elicited a predominantly M1 type response and resulted in deposition of dense connective tissue and/or scarring. We conclude that the presence of cellular material within an ECM scaffold modulates the phenotype of the macrophages participating in the host response following implantation, and that the phenotype of the macrophages participating in the host response appears to be related to tissue remodeling outcome. PMID- 19121542 TI - Reproductive glycogenetics--a critical factor in pregnancy success and species hybridisation. AB - Hybridisation occurs rarely in nature and experiments using interspecific transfer of embryos generally result in implantation failure. Here we show that appropriate glycosylation of the apposing surfaces of endometrium and trophoblast probably is an important factor and may play a critical role in pregnancy success. Examination of closely related species shows that each has its own specific pattern of glycosylation, or glycotype, at the fetomaternal interface and that interacting surfaces appear to show complementarity, suggesting the existence of a glycocode. Studies on a camel/llama hybrid show that for successful implantation to occur, a hybrid must have a placental glycosylation pattern similar to that of the host species, suggesting that the glycocode and appropriate glycosylation may be critical factors in the establishment and maintenance of pregnancy. This new field of reproductive glycogenetics is not only relevant to the development of new species but may also have important implications in the area of human fertility. PMID- 19121541 TI - A modification of the uterine artery restriction technique in the guinea pig fetus produces asymmetrical ultrasound growth. AB - AIMS: To evaluate the use of diathermy ablation of branches of the uterine artery to produce growth restriction in the fetal guinea pig, and to compare this new approach with the more conventional use of uterine artery ligation. The development of growth restriction was documented by measuring fetal biparietal diameter (BPD) and the resistance index (RI) of the umbilical artery blood flow velocity waveform. METHODS: At 30-35 days of gestation (term=70 days), one uterine artery was ligated in 29 sows. In another 16 sows, branches of one uterine artery were ablated using diathermy. Fetuses in contralateral horns were used as controls. Ultrasound measurements were made weekly, and at 59-69 days of gestation animals were euthanased to determine fetal position in utero as well as fetal and placental weights. In some fetuses, brain and liver weights were also recorded. RESULTS: Both surgical techniques resulted in similar reductions in fetal body and placental weights. The number of fetuses surviving to term was greater in the diathermy group (53%) compared to the ligation group (22%) (P<0.05). Results from these two groups were combined and referred to as "treated" fetuses. The brain/liver weight ratio was increased by 245% in the treated fetuses compared to control fetuses. Ultrasound measures of BPD in the treated fetuses were within the normal range. The mean RI of the treated group showed a slight but significant increase near term compared to the mean RI of the normal range. CONCLUSION: We have shown that the diathermy technique produces asymmetrical fetal growth restriction (with normal head size) in the guinea pig to the same extent as the conventional ligation technique. It is associated with a lower fetal mortality rate and therefore should be the preferred method. The minimal increase in umbilical artery resistance index only at the end of gestation amongst the intrauterine growth restriction (IUGR) fetuses suggests that an obliterative vasculopathy in the umbilical circulation is not the cause of growth failure when there is maternal uteroplacental restriction. PMID- 19121540 TI - The influence of Ni(II) on surface antigen expression in murine macrophages. AB - Biomedical alloys may release nickel ions during corrosion phenomena and, in addition to their interaction with oral tissues, these ions may also influence characteristic properties of the immune system cells. The aim of this study was to evaluate the effect of nickel chloride on the expression of functionally distinct surface antigens in murine RAW macrophages. The expression of the surface antigens CD14, CD40, MHC class I, MHC class II, CD80, CD86, CD54 was analyzed by flow cytometry. The bacterial endotoxin lipopolysaccharide (LPS) was used as a positive control to induce antigen expression. Cells were stimulated with NiCl(2) (0.1 and 0.5mm) in the presence and absence of LPS (0.1 or 25 microg/ml). After exposure periods of 6, 24 and 48 h, LPS caused a time- and dose dependent increase in the expression of all surface antigens. CD14 expression was up-regulated by 0.1 microg/ml LPS by about 10-fold after 24h and 100-fold after 48 h. After 48 h, NiCl(2) alone up-regulated the expression of all surface antigens between 2- and 4-fold, while in cells stimulated by LPS, 0.1mm NiCl(2) was effective only on CD14, CD40 and MHC class I. Moreover, 0.5mm NiCl(2) even inhibited the LPS-induced expression of all surface antigens, except for CD54, which was still significantly up-regulated. These results show that nickel chloride is able to induce an up-regulation of surface antigen expression, but a high concentration may impair essential functions of macrophages stimulated by LPS. PMID- 19121545 TI - Missed opportunities to diagnose Plasmodium falciparum malaria: results of a regional service evaluation. PMID- 19121543 TI - Anionic PAMAM dendrimers as drug delivery vehicles for transition metal-based anticancer drugs. AB - The use of anionic half-generation poly(amidoamine) dendrimers as drug delivery vehicles for [Pt(S,S-dach)(5,6-Me(2)phen)](2+) (56MESS) (where S,S-dach=1S,2S diaminocyclohexane; 5,6-Me(2)phen=5,6-dimethyl-1,10-phenanthroline) and [{Delta,Delta-Ru(phen)(2)}(2)(mu-bb7)](4+) (Rubb(7)) (where phen=1,10 phenanthroline; bb7=1,7-bis[4-(4'-methyl-2,2'-bipyridyl)heptane]) has been studied by nuclear magnetic resonance spectroscopy. From one- and two-dimensional (1)H NMR spectra both 56MESS and Rubb(7) were found to bind to the surface of generation 3.5, 4.5, 5.5 and 6.5 dendrimers through electrostatic interactions. The higher charge and larger size of Rubb(7) resulted in stronger binding to all dendrimer generations (K(b)> or =2 x 10(5)M(-1)) compared with 56MESS (K(b)> or =1 x 10(4)M(-1)). Interestingly, there appeared to be no observable trend between dendrimer size and binding constant strength. The size of the free and 56MESS bound dendrimers were examined using pulsed-gradient spin-echo NMR. The dendrimers ranged in hydrodynamic diameter from 11 to 20 nm and in all cases were larger than their corresponding full-generation dendrimer. Upon the addition of 56MESS the diameter of the dendrimers increased, consistent with surface binding. PMID- 19121544 TI - Aminoacid-derivatised picolinato-oxidovanadium(IV) complexes: characterisation, speciation and ex vivo insulin-mimetic potential. AB - The proligands PicMe-AaR (PicMe=methoxipicolyl-5-amide, where the amide substituent is an amino acid AaR=HisH, HisMe, IleH, IleMe, TrpH, TrpMe, HTyrEt, tBuTyrMe, HThrMe, tBuThrMe) and the complexes [VO(Pic-AaR)(2)] have been synthesised and characterised. A detailed EPR study of the VO(2+)/Pic-His systems in water revealed the predominance of the complex [VO(Pic-His)H(2)O] in the pH range 2-6, with tridentate coordination of Pic-His via the picolinate moiety and imidazole-Ndelta. Speciation analyses of the binary systems VO(2+)/Pic-Aa (Aa=His, Ile, Trp) and the ternary systems VO(2+)/Pic-Aa/B (Aa=His, Ile; B=citrate (cit), lactate (lac), phosphate) showed a predominance of the ternary complexes [VO(Pic-Aa)(cit/lac)] and [VO(Pic-Aa)(cit/lac)OH](-) in the physiological pH regime. If, in addition, human serum albumin (HAS) and apotransferrin (Tf) are present, with all of the low and high molecular mass constituents in their blood serum concentrations, about two thirds of VO(2+) is bound to the protein, while there is still a sizable amount of ternary complex [VO(Pic-Aa)(cit/lac)] present (about 1/4 for Pic-His and 1/3 for Pic-Ile) when the vanadium(IV) concentration is relatively high; at lower concentrations Tf is the predominant binder. Insulin-mimetic studies for VO(2+)/Pic-Aa (Aa=His, Ile, Tyr and Trp), based on a lipolysis assay with rat adipocytes, provided IC(50) values of 0.41(1) for VO(2+)/Pic-His and VO(2+)/Pic-Ile, which compares with 0.87(17) for VOSO(4). PMID- 19121546 TI - Suicide risk in rapid cycling bipolar patients. AB - BACKGROUND: Rapid-cycling (RC) is a course modifier of bipolar disorder which often implicates a poor prognosis. However, the relationship of RC with suicidal features as a marker of impairment has been understudied. METHODS: Three hundred and five patients (n=305) were included in a naturalistic, systematic prospective study in a single site setting. Patients with rapid-cycling (RC) were defined as having four or more manic, mixed or depressive episodes in the year prior to baseline assessment. The two groups were compared regarding clinical and sociodemographic variables, paying special attention to suicidal features. Statistical methods consisted of chi-square statistic for the comparison of categorical data, and Student's t test for dimensional variables normally distributed. Also, a General Linear Model was used to study the main effect of different sociodemographic and clinical variables on suicidality. All statistics were two-tailed, and significance was set at p less than 0.05. RESULTS: Fifty five patients (18%) were classified as RC, whilst 250 (82%) were considered as nonrapid-cycling (NRC). No significative difference was found in the prevalence of RC amongst bipolar I and II patients. RC was associated with depressive onset of bipolar disorder. The number of suicide attempts was significantly higher amongst RC (RC mean 0.82 [SD 1.85] vs. NRC 0.44[SD 0.94] t=2.09, p=0.37). Nonetheless, no significative differences were found between RC and NRC regarding the percentage of suicide attempters. On the other hand patients that presented RC showed a marked increase of lifetime history of suicidal ideation (Chi(2)=4.363, p=0.039). Finally, there were not any differences between RC patients and NRC in family history of suicide. DISCUSSION: Bipolar patients with RC are more likely to attempt suicide. Intensive treatment of this marker should be considered. PMID- 19121547 TI - Prostate delineation using CT and MRI for radiotherapy patients with bilateral hip prostheses. AB - BACKGROUND AND PURPOSE: Prostate cancer patients with bilateral hip prostheses (b HP) are rare, but pose unique challenges for high-dose radiotherapy. The purpose of this study was to determine the effect of b-HP on prostate delineation using CT and MR. METHODS AND MATERIALS: The planning CT and MR of 7 patients with b-HP were co-registered. Four observers delineated the prostate on both data sets, blinded to the volumes delineated by their peers and those on the alternate image set. Inter-modality differences and inter-observer variability were quantified by calculating 3D volume overlaps. The effect of prostheses physical extent on these variables was evaluated. MR geometric distortion was explored using the differences in vector distances between three intra-prostatic markers (IPMs) on CT and MR. RESULTS: Prostate volumes on CT were consistently larger than MR volumes in all patients when averaged over the 4 observers (+10% to +46%, p<0.001). The mean inter-modality volume overlap was 1.59, which varied significantly between patients (1.35 to 1.82, p=0.04). There was a trend toward smaller inter-observer variability in the MR volumes (1.95 vs. 1.71, p=0.08). No differences could be identified between the IPM vectors on CT and MR. CONCLUSIONS: Although limited by a small sample size, this study found that MR prostate volumes were significantly smaller than those from CT for b-HP patients. The largest differences between CT and MR prostate volumes and the greatest inter observer variability on CT tended to be seen in patients with the largest prostheses. b-HP do not appear to cause measurable MR geometric distortions in the sequences used. PMID- 19121548 TI - Predicting the anticipated emotional and behavioral responses to an avian flu outbreak. AB - BACKGROUND: The purpose of this study was to develop a model to predict the emotional and behavioral responses to an avian flu outbreak. METHODS: The participants were 289 university students ranging in age, income, and ethnic backgrounds. They were presented with scenarios describing avian flu outbreaks affecting their community. They reported their anticipated emotional responses (positive emotion, negative emotion) and behavioral responses (helping, avoidance, sacrifice, illegal behavior) as if the scenarios were actually occurring. They also were assessed on individual differences expected to predict their responses. RESULTS: Participants were only modestly familiar with the avian flu and anticipated strong emotional and behavioral responses to an outbreak. Path analyses were conducted to test a model for predicting responses. The model showed that age, sex, income, spirituality, resilience, and neuroticism were related to responses. Spirituality, resilience, and income predicted better emotional responses, and neuroticism and female sex predicted worse emotional responses. Age, sex, income, and spirituality had direct effects on behavior. The emotional responses were directly related to each behavior and mediated the effects of individual differences. CONCLUSION: Emotional responses may be important in predicting behavior after an outbreak of avian flu, and personal characteristics may predict both emotional and behavioral responses. PMID- 19121549 TI - Update on infection prevention in disaster planning: new resources and policies. PMID- 19121551 TI - Supporting enrolled nurse conversion--the impact of a compulsory bridging program. AB - In Australia, unlike the UK, two levels of nursing registration have been retained. In developing a course for students to convert from level two to level one of the register, the need for bridging biological science was identified. The one-week course focused on learning strategies and familiarisation with resources in science and aimed to support students to make the transition to university level study. An evaluation of the program examined students' perception of confidence, changes in their learning skills, and the impact of the bridging program on ongoing performance in their degree. Using questionnaires, confidence levels in returning to study, confidence levels in the biological sciences, the initiation of study contacts and the overall impact of the bridging program on university transition were evaluated. Results indicated a 51% increase in overall return to study confidence levels and a 63% increase in confidence with the biological sciences. Students indicated that the bridging program assisted their coping by being exposed to a range of methods, styles and approaches and they had the opportunity to build study contacts. While our research has implications for conversion programs, there are wider implications for the transition of all mature age students to the university sector. PMID- 19121550 TI - Design, synthesis and hypolipidemic activity of novel 2-(m-tolyloxy) isobutyric acid derivatives. AB - Novel 2-substituted isobutyric acid derivatives were synthesized and their hypolipidemic activity was evaluated in high cholesterol diet fed rat model. The amide 5a was found to decrease the levels of serum total cholesterol, LDL cholesterol and triglycerides in hyperlipidemic rats to a greater degree than the reference gemfibrozil. PMID- 19121552 TI - Pregnancy health status of sub-Saharan refugee women who have resettled in developed countries: a review of the literature. AB - OBJECTIVE: to present the literature relating to health status and pregnancy complications among sub-Saharan African women. BACKGROUND: sub-Saharan refugee women constitute a new and growing group of maternity service users in developed countries today. These women are perceived to be at high risk of pregnancy complication, based on concurrent disease and unusual medical conditions. As a result of these concerns, midwives may feel ill equipped to provide their pregnancy care. METHOD: searches were conducted of CINAHL, Maternity and Infant Care, MEDLINE and PsychINFO databases using the search terms 'migrants', 'Africa', 'sub-Saharan', 'pregnancy', 'refugees' and 'women'. Additional articles were located by pursuing references identified in key papers. FINDINGS: pregnant sub-Saharan women present as an at-risk population related to poor prior health, co-existing disease and cultural practices such as female genital mutilation. Nonetheless, principal pregnancy complications for this population include anaemia and high parity, rather than exotic disease. Higher rates of infant mortality and morbidity appear to persist following resettlement, and are not explained by maternal risk factors alone. Limited access to care is of concern. KEY CONCLUSIONS: further research is warranted into the impediments to care uptake among sub-Saharan African women. It is hoped that such research will inform the development of culturally appropriate and acceptable services for African refugees. IMPLICATIONS FOR PRACTICE: it is important that midwives are aware of common health problems among sub-Saharan women. Midwives also need to act to promote access to health services among this group. Social disadvantage and late access to care may impact on neonatal outcomes and thus warrant investigation. PMID- 19121553 TI - Evaluation and in situ assessment of photodegradation of polyaromatic hydrocarbons in semipermeable membrane devices deployed in ocean water. AB - Semipermeable membrane devices (SPMDs) were deployed in water using four different methods: a typical SPMD cage with and without a mesh cover, a bowl chamber and without any protection. In addition to routinely used performance reference compounds (PRCs), perdeuterated dibenz[a,h]anthracene was added. Due to its high sampler to water partition coefficient no measurable clearance due to diffusion was expected during the deployment period, hence any observed loss could be attributed to photodegradation. The loss of PRCs was measured and SPMD based water concentrations determined. Results showed that a typical SPMD deployment cage covered with mesh provided the best protection from photodegradation. Samplers which had undergone the highest photodegradation underestimated PAH water concentrations by up to a factor of 5 compared to the most protected SPMDs. This study demonstrates that the potential for photodegradation needs to be addressed when samplers are deployed in water of low turbidity. PMID- 19121554 TI - Mercury fluxes and pools in three subtropical forested catchments, southwest China. AB - Forested catchments are an important part of the mercury (Hg) cycle and a link between the atmospheric and the aquatic environments. In this study, Hg input and output fluxes and its retention were investigated at subtropical forested catchments in southwest China. Significantly enhanced atmospheric Hg inputs were observed, and the contribution of litterfall Hg plays a more important role at these subtropical forested catchments. The ratios of Hg output fluxes from stream water to total input were 2.5% and 1.2% for LGS and TSP, which were markedly lower than those reported from Europe and North America. The current annual input Hg only accounted for 0.8 and 1.8 per mille to the Hg stored in the upper 90 cm of soil in LGS and TSP. These suggest that subtropical forested catchments are important for retention of atmospheric mercury deposition in southwest China. PMID- 19121555 TI - Crescentic glomerulonephritis with ribbon-like immunofluorescence pattern. PMID- 19121556 TI - Subacute allograft failure with dysuria and hematuria in a kidney transplant recipient. PMID- 19121557 TI - Medium-dependent interactions of quinones with cytosine and cytidine: a laser flash photolysis study with magnetic field effect. AB - Laser flash photolysis and an external magnetic field have been used for the study of the interaction of two quinone molecules, namely, 9,10-anthraquinone (AQ) and 2-methyl 1,4-naphthoquinone (or menadione, MQ) with a DNA base, cytosine (C) and its nucleoside cytidine (dC) in two media, a homogeneous one composed of acetonitrile/water (ACN/H(2)O, 9:1, v/v) and a SDS micellar heterogeneous one. We have applied an external magnetic field for the proper identification of the transients formed during the interactions in micellar media. Cytosine exhibits electron transfer (ET) followed by hydrogen abstraction (HA) while dC reveals a reduced ET compared to C, with both quinones in organic homogeneous medium (ACN/H(2)O). Due to a higher electron affinity, AQ supports more faciler ET than MQ with dC in ACN/H(2)O but observations in SDS have been just the reverse. In SDS, ET from dC is completely quenched and a dominant HA is all that could be discerned. This work reveals two main findings: first, a drop in ET on addition of a ribose unit to C, which has been attributed to a role of keto-enol tautomerism in inducing ET from electron-rich nucleus and second, the effect of medium in controlling reaction mechanism by favoring HA with AQ although it is intrinsically more prone towards ET. PMID- 19121558 TI - Perceived intensity and unpleasantness of cutaneous and auditory stimuli: an evaluation of the generalized hypervigilance hypothesis. AB - According to the Generalized Hypervigilance Hypothesis (GHH) of McDermid et al., the unpleasantness of sensory stimuli, rather than their modality, determines whether they will be perceptually amplified in hypervigilant persons. In a test of this idea, ratings of the intensity of sensations evoked by cutaneous and auditory stimuli were obtained from individuals with chronic myofascial pain (fibromyalgia, temporomandibular disorders), and from (less hypervigilant) healthy control participants. In each modality, the stimuli spanned a wide intensity range, with the weakest stimuli being affectively neutral and the strongest being distinctly unpleasant, as determined by unpleasantness ratings. The pain patients showed robust perceptual amplification of the cutaneous pressure stimuli, and modest amplification of the auditory stimuli. In both cases, perceptual amplification extended to even the lowest stimulus intensities, a result that is not consistent with the predictions of the GHH. An alternative formulation, the attentional gain control model of hypervigilance, is proposed, according to which those types of stimuli that are associated with pain are amplified because of the attention that is habitually directed toward them. PMID- 19121559 TI - Skin regeneration: What determines it? PMID- 19121560 TI - Cardiac asystole at birth: Is hypovolemic shock the cause? AB - A birth involving shoulder dystocia can rapidly deteriorate-from a fetus with a reassuring tracing in the minutes before birth, to a neonate needing aggressive resuscitation. Infants experiencing a traumatic birth involving shoulder dystocia may be severely compromised, even when the preceding labor was uncomplicated. This paper presents two cases in which infants had normal heart beats recorded 5 10min before birth and were born with cardiac asystole following shoulder dystocia. Often, in cases of shoulder dystocia, infants shift blood to the placenta due to the tight compressive squeeze of the body in the birth canal (along with cord compression) and thereby may be born hypovolemic. Our hypothesis is that the occurrence of sudden cardiac asystole at birth is due to extreme hypovolemic shock secondary to the loss of blood. At birth, the sudden release of pressure on the infant's body results in hypoperfusion resulting in low central circulation and blood pressure. Severe hypovolemic shock from these effects leads to sudden cardiac arrest. Immediate cord clamping maintains the hypovolemic state by preventing the physiologic and readily available placental blood from returning to the infant. Loss of this blood initiates an inflammatory response leading to seizures, hypoxic-ischemic encephalopathy, and brain damage or death. Animal studies have shown that human umbilical stem cells injected into a rat's abdomen after induced brain damage, can protect the rat's brain from developing permanent injury. To prevent damage to newborns, the infant must receive the blood volume and stem cells lost at the time of descent and immediate cord clamping. Recommended countermeasures for research include: (1) resuscitation at the perineum with intact cord; or (2) milking the cord before clamping; or (3) autologous transfusion of placenta blood after the birth; or (4) rapid transfusion of O negative blood after birth and before seizures begin. PMID- 19121561 TI - Use of fluoroscopy in determining screw overshoot in the dorsal distal radius: a cadaveric study. AB - PURPOSE: To determine the ability of standard fluoroscopic imaging to assess whether screws placed from volar to dorsal during distal radius plating have penetrated the dorsal cortex of the radius. METHODS: Hard-copy digital images of cadaveric distal radii with screws of different lengths penetrating the dorsal cortex of the radius were shown to practitioners. The group included attending hand surgeons and hand fellows in training at multiple institutions. The participants were then asked to determine, from multiple images presented, whether the screws penetrated the dorsal cortex of the radius or were acceptable in length. RESULTS: Much more accurate assessment of screw positioning occurred with radial screw positions. Experienced practitioners showed higher accuracy when evaluating screws in any position. Both groups showed the least accuracy in evaluating screws in the ulnar column of the radius. For all evaluators, fluoroscopy was found to be 82% sensitive at detecting cortical penetration in the radial-most position, 77% sensitive in the central position, and 57% sensitive in the ulnar position. Evaluators with >3 years in practice were able to detect incorrect screw positions with 100% sensitivity in the radial-most position, 90% in the central position, and 75% in the ulnar position. Inexperienced evaluators detected incorrect screw positioning with 70% sensitivity in the radial position, 61% centrally, and 56% ulnarly. CONCLUSIONS: Accurate placement of screws used with volar fixation devices is important to avoid prominent hardware, especially screws that may penetrate dorsally into tendon compartments. Printed image intensifier images have limited sensitivity for the diagnosis of dorsal cortical penetration of a volarly inserted screw, particularly among less experienced observers and for the evaluation of the most ulnar screw positions. PMID- 19121562 TI - Osteochondral autografting for acute articular defect of the metacarpal head: case report. AB - Osteochondral injuries in the hand are common. The small size of the joints and associated articular surfaces make repair of these injuries difficult. Osteochondral autografting has been described in the knee for some time and has recently been reported for hand injuries. We describe the use of an osteochondral graft taken from the dorsal nonarticular portion of the metacarpal head for a defect in the metacarpophalangeal joint surface. PMID- 19121563 TI - Early outcome of spherical ceramic trapezial-metacarpal arthroplasty. AB - PURPOSE: A retrospective review of 50 ceramic, spherical implants for the treatment of trapezial-metacarpal osteoarthritis in 49 patients assessed clinical and radiographic outcome. METHODS: Thirty-five patients returned for a clinical examination and completed an outcome survey at a mean follow-up of 3 years. A radiographic review was available for all 50 implants with a mean follow-up of 3 years. Average patient age at surgery was 59 years (35 women and 15 men). Twenty five patients were retired and 24 were regularly employed preoperatively. RESULTS: Overall satisfaction was between "very satisfied" and "satisfied" for 32 patients, and none were dissatisfied. Twenty-nine patients rated both frequency and severity of pain as "less than before surgery" or "much less than before surgery." Average rating for thumb and hand strength was the "same as before surgery." Thumb pinch strength averaged 40 N (91% that of the opposite thumb). Postoperative thumb opposition was Kapandji stage 8 or better in 34 of 50 thumbs and stage 6 or better in patients who had an interphalangeal joint or metacarpophalangeal joint fusion. All 24 employed patients returned to work, but 2 required substantial modifications of pinch and grasp movements. Implant subsidence occurred in 47 of 50 thumbs, ranging from 1 to 13 mm. Trapezium fractures occurred in 15 thumbs. Eleven additional thumbs showed erosions and cystic changes within the trapezium or metacarpal. There was no evidence of arthritis developing in surrounding joints. In addition to adverse radiographic changes, there were 10 complications in 10 patients, including implant subluxation in 7 patients. CONCLUSIONS: Although most patients were satisfied with their function and pain relief, adverse radiographic findings were nearly universal. Implant subsidence, especially into the trapezium, was often severe, with some resulting in a trapezium fracture. Based on the radiographic outcome at this medium-term follow-up, we no longer use this implant. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 19121564 TI - Obesity and risk of the less commonly diagnosed subtypes of breast cancer. AB - OBJECTIVES: A set of common epidemiologic risk factors have been associated with the risk of breast cancer despite of its molecular sub-classifications. We implemented a case series study with the primary objective of evaluating if obesity is associated with the diagnostic risk of "ER+ and/or PR+, HER2+", "ER /PR-, HER2-", or "ER-/PR-, HER2+" relative to the most commonly diagnosed subtype of breast carcinoma, "ER+ and/or PR+, HER2-". METHODS: Demographic, clinical and pathologic data were collected from existing databases. The statuses of HER2/neu biomarker and hormone receptors were dichotomized as either positive or negative. Immunohistochemical staining was used to assess the prevalence of different subtypes. Body mass index was calculated from weight and height data collected at the time of consultation. CONCLUSIONS: Findings from the present study suggest that excess body weight decreases the diagnostic risk of "ER-/PR-, HER2-", or "ER /PR-, HER2-" relative to "ER+ and/or PR+, HER2-". Obese and overweight women are more likely to be diagnosed with to "ER+ and/or PR+, HER2-", the subtype that has best prognosis and mostly associated with personal lifestyle. Weight gain with the population attributable-risk factor of 21.3% contributes the most to the incidence of invasive post menopausal breast cancer. Younger pre-menopausal women were more likely to be diagnosed with "ER+ and/or PR+, HER2+". In younger women biology of breast cancers with positive expression for hormone receptors and epidermal growth factor is a complex that extends beyond the currently assessed prognostic markers. PMID- 19121565 TI - Learning and generation of goal-directed arm reaching from scratch. AB - In this paper, we propose a computational model for arm reaching control and learning. Our model describes not only the mechanism of motor control but also that of learning. Although several motor control models have been proposed to explain the control mechanism underlying well-trained arm reaching movements, it has not been fully considered how the central nervous system (CNS) learns to control our body. One of the great abilities of the CNS is that it can learn by itself how to control our body to execute required tasks. Our model is designed to improve the performance of control in a trial-and-error manner which is commonly seen in human's motor skill learning. In this paper, we focus on a reaching task in the sagittal plane and show that our model can learn and generate accurate reaching toward various target points without prior knowledge of arm dynamics. Furthermore, by comparing the movement trajectories with those made by human subjects, we show that our model can reproduce human-like reaching motions without specifying desired trajectories. PMID- 19121566 TI - Regional myocardial function after myocardial infarction in mice: a follow-up study by strain rate imaging. AB - BACKGROUND: New therapeutic strategies aim to reduce the extent of myocardial infarction (MI) to decrease long-term left ventricular (LV) remodeling. These innovations are often developed on murine models of MI and have led to the need for a sensitive tool allowing follow-up. The aim of this study was to investigate by strain rate (SR) imaging early and long-term alteration in regional LV function occurring after MI in mice. METHODS: Echocardiography was serially performed during a 4-month follow-up period in 3 groups of C57BL6 male mice: 7 normal, 5 sham operated, and 27 with left coronary artery ligation (the MI group). In addition to conventional measurements, SR was obtained from short-axis views in the anterior wall and posterior wall (PW). Triphenyltetrazolium chloride staining allowed the localization and measurement of the transmural extent of MI. A transmural MI was defined as an extension > 75% of the wall thickness. RESULTS: In the MI group, LV ejection fractions significantly decreased, while LV dimensions and PW thicknesses increased from baseline to 4 months. On day 3, SR could differentiate transmural from nontransmural (1 +/- 1 vs 10 +/- 1 s(-1); P < .05) and noninfarcted myocardium (25 +/- 1 s(-1)). SR values did not significantly change between day 3 and month 4 and could still differentiate those segments at month 4. Wall thickening was not able to differentiate transmural from nontransmural infarcted segments at day 3 (16 +/- 3% vs 21 +/- 3%; P = NS) or at month 4. CONCLUSION: In this murine model of MI, SR was able to predict the transmural extent of MI as early as 3 days after MI, then remained stable and still differentiated them at 4 months. PMID- 19121567 TI - Feasibility of prediction of myocardial viability with Doppler tissue imaging following percutaneous coronary intervention for ST elevation anterior myocardial infarction. AB - BACKGROUND: In patients with acute ST elevation myocardial infarction (STEMI), it is clinically important to determine the viability of akinetic segments soon after acute reperfusion therapy. The purpose of this study was to determine whether Doppler tissue imaging can predict myocardial viability in this clinical setting. METHODS: Twenty-four consecutive patients with the first acute anterior STEMI with akinetic apical segments were enrolled. Color-coded Doppler tissue imaging was performed. Myocardial velocity and strain values were determined from the septal and lateral walls at the normal basal and akinetic apical levels of the left ventricle on day 1 after percutaneous coronary intervention. The presence of isovolumic contraction of tissue velocity (TVivc) and strain rate (SRivc) were also determined. RESULTS: Twenty patients (mean age 62 +/- 15 years; 11 men) returned for follow-up echocardiography to assess wall motion recovery and viability. Ten patients who had recovery of akinetic segments showed lower baseline E/e' ratios than those without recovery (13.4 +/- 5.9 vs 19.1 +/- 5.7; P = .04). There was no difference between 19 recovered and 21 nonrecovered apical segments in all TV, SR, and strain values except early diastolic SR (SRe; 0.64 +/ 0.35 vs 0.43 +/- 0.25 s(-1); P = .04) at day 1. With a cutoff value of 0.32 s( 1) for SRe, the receiver operating characteristic curve for the prediction of recovery showed the highest sensitivity of 84%. The presence of TVivc had sensitivity and specificity of 79% and 33%, respectively, and for SRivc, sensitivity and specificity were 84% and 63%, respectively. With the combination of SRe and SRivc, specificity was increased to 78%. CONCLUSION: Patients with functional recovery from anterior STEMI showed better diastolic function, better SRe, and more isovolumic contraction. These parameters appear to be promising predictors for myocardial viability, and SR imaging was found to be a better method than TV imaging for the identification of viable myocardium in patients with STEMIs who underwent percutaneous coronary intervention. PMID- 19121568 TI - Doppler myocardial imaging compared to standard two-dimensional and Doppler echocardiography for assessment of diastolic function in patients with systemic amyloidosis. AB - BACKGROUND AND AIMS: Cardiac amyloidosis (AL) is characterized by early impairment of diastolic function with preservation of systolic function, as assessed by standard measures, until very late in the disease process> Role of diastolic color Doppler myocardial imaging (cDMI), that is diastolic color Doppler myocardial velocity imaging (MVI) and strain rate imaging (SRI), to assess distolic dysfunction in this population remains unclear. METHODS: 238 patients with biopsy proven systemic primary (AL) amyloidosis and 39 age and sex matched controls have been enrolled. Left ventricular diastolic cDMI measures were compared in patients and healthy subjects, to test if such diastolic dysfunction occurs before longitudinal left ventricular systolic dysfunction. RESULTS: Compared to Diastolic longitudinal, radial, and circumferential DMI, standard pulsed wave tissue Doppler imaging (PW-TDI) of the mitral annulus was the most accurate technique to detect early diastolic dysfunction in patients with AL. However, systolic longitudinal peak cDMI measures outperformed all of the diastolic measures, including PW-TDI, in separating patients with systemic AL, but no evidence of cardiac involvement on standard 2D and Doppler evaluation, from controls. CONCLUSIONS: Even when compared to diastolic cDMI measurements, standard PW-TDI of the mitral annulus was the most accurate diastolic measure to detect early left ventricular dysfunction in patients with AL amyloidosis. However, no diastolic measure approached the accuracy of longitudinal systolic cDMI measurements in identifying ventricular dysfunction in AL patients with normal standard 2D and Doppler examinations, compared to controls. PMID- 19121569 TI - A comparison of Tei index versus systolic to diastolic ratio to detect left ventricular dysfunction in pediatric patients. AB - BACKGROUND: The aim of this study was to determine the accuracy, sensitivity, specificity, predictive values, and likelihood ratios of the left ventricular (LV) Tei index (TX) and the ratio of systolic duration to diastolic duration (S/D) to detect ventricular dysfunction. METHODS: LV systolic and diastolic function were studied in a cohort of 68 children, 25 normal and 43 abnormal. Systolic dysfunction was defined as the presence of all of 3 criteria: ejection fraction < 50%, fractional shortening < 27%, and tissue Doppler systolic S wave < 1.5 standard deviations below normal (Z-1.5). Diastolic dysfunction was defined as the presence of all of 3 criteria: isovolumic relaxation time corrected for heart rate > 88 ms, ratio of transmitral Doppler flow early diastolic velocity (E) to mitral valve annular early diastolic tissue Doppler velocity (E') > 13, and E' < Z-1.5. The cohort was divided into 4 groups: normal systolic and diastolic function, systolic dysfunction, diastolic dysfunction, and combined systolic and diastolic dysfunction. TX > 0.5 and S/D > 1.0 were defined as abnormal. TX and S/D results were compared with normal function versus abnormal function categories using 2 x 2 classification tables and receiver operating characteristic curves. RESULTS: The overall accuracy of TX was 85%, and that of S/D was 82%. TX was most accurate (92%) for detecting diastolic and combined dysfunction, whereas S/D had similar accuracy for all LV function groups. The overall accuracy, sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio of TX were slightly greater than those for S/D for almost all categories. CONCLUSION: The overall accuracy, sensitivity, specificity, predictive values, and likelihood ratios of TX are slightly greater than those of S/D for most categories of dysfunction, although either may fail to detect dysfunction in 15% to 18% of cases. When both TX and S/D are used in combination, using the presence of either TX > 0.5 or S/D > 1.0 as indicative of dysfunction, few cases of LV dysfunction should elude detection. PMID- 19121570 TI - The effect of ulinastatin on the small intestine injury and mast cell degranulation in a rat model of sepsis induced by CLP. AB - INTRODUCTION: Sepsis could be initiated by the gastrointestinal tract injury and subsequent bacterial translocation. In the present experiment, we aimed to investigate effect of ulinastatin (UTI) on the small intestinal injury and bacterial translocation in septic rats and role of mast cells degranulation in its action. METHODS: Fifty-four male Wistar rats were randomly divided into three groups: sham laparatomy, cecal ligation and puncture (CLP), and CLP plus UTI. CLP was used to develop septic rat model and UTI was administered to rats intraperitoneally (50,000 U/kg) 30 min prior to CLP operation. After CLP or sham operation, variable parameters were investigated in three subsets of animals. One subset was used for measurements of nitrite and nitrate (NO(x)) concentration in plasma at 1, 6, 12, 18, and 24h and levels of NO(x) and iNOS mRNA in the small intestine, RMCP-II released into the small intestinal lumen, bacterial translocation and morphologic changes at 24h. The other subsets were used for the small intestinal motility and microvascular in vivo at 24h. RESULTS: Bacterial translocation, barrier injury, impaired motility and blood flow, mast cells degranulation of the small intestine in the CLP group were found more severe than that in the sham group. Elevated RMCP-II, NO(x), and iNOS mRNA levels were also detected in the CLP group. Application of UTI not only protected the small intestine from sepsis but also diminished changes of intestinal mast cells. CONCLUSION: UTI can significantly ameliorate the small intestinal injury and subsequent bacterial translocation by inhibiting mast cells degranulation in septic rats. PMID- 19121571 TI - Influence of cell culture media conditions on the osteogenic differentiation of cord blood-derived mesenchymal stem cells. AB - In this study the critical parameters directing osteogenic differentiation of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) were investigated, key factors and conditions identified and improved protocols for a more cell-type adapted differentiation developed. Today only little information about the specific conditions directing osteogenic development is available and current protocols for cultivation and differentiation of UCB-MSCs are based mainly on experience with bone marrow-derived MSCs (BM-MSCs) without further adaptation. Thus, protocols for improved osteoinduction are of particular interest. The goal of this study was to investigate the influence of three different culture media (A) alpha MEM, 15% FBS, (B) DMEM, 15% FBS and (C) MSCGM, 10% SingleQuot growth supplement on the osteogenic differentiation of UCB-MSCs. Moreover, a systematic analysis of two concentrations of dexamethasone (10(-8)M/10(-7)M) in combination with or without BMP-2 (10(-7)M) was carried out by detecting the expression of alkaline phosphatase (ALP), collagen-1 and the mineralization of ECM. We found that MSCGM, 10% SingleQuot had a supportive effect on the osteogenic differentiation of UCB-MSCs. In case of treatment with 10(-8)M dexamethasone, mineralization occurred in combination with BMP-2 exclusively, while a concentration of 10(-7)M dexamethasone led to a high amount of mineralized ECM and the expression of collagen-1 independent of BMP-2 addition. According to this data dexamethasone is the leading osteoinductive factor, but BMP-2 seems to have supportive properties in UCB-MSCs. In conclusion, MSCGM supplemented with 10% SingleQuot and 10(-7)M dexamethasone was the condition identified to be best for inducing the osteogenic differentiation of UCB-MSCs. PMID- 19121572 TI - Pharmacodynamic action and mechanism of volatile oil from Rhizoma Ligustici Chuanxiong Hort. on treating headache. AB - The volatile oil from Rhizoma Ligustici Chuanxiong Hort. (CXVO) is likely to be the mainly active ingredient of Chuangxiong in curing headache. In this study, oral administration of CXVO (45.0, 90.0, and 135.0 microl/kg) to mice significantly elevated the pain threshold in the hot-plate test and reduced the number of abdominal writhing caused by acetic acid. CXVO (90.0 and 135.0 microl/kg) not only reduced locomotor activity, but also prolonged the sleeping time induced by sodium pentobarbital (35 mg/kg), and the number of mice with sleeping time over 1 min by sodium pentobarbital (25 mg/kg) was markedly enlarged by CXVO (45.0, 90.0, 135.0 microl/kg) administration. The three doses of CXVO significantly increased the pain threshold of rabbits with headache due to hot radiation and the level of plasma ET of rats with headache due to nitroglycerin injection. Besides, for the nitroglycerin-induced headache rats, the c-fos gene expression in the brain stem and hypothalamus was remarkably inhibited and the level of plasma CGRP was reduced significantly after CXVO administration at both doses 90.0 and 135.0 microg/kg. The latter dosage could also raise the level of plasma 5-HT markedly. The study suggests that CXVO acts probably as the active ingredient of Rhizoma Ligustici Chuanxiong Hort. (CX) on treating headache and has potential to be an agent for treating headache. PMID- 19121573 TI - Inhibition of cough-reflex sensitivity by benzonatate and guaifenesin in acute viral cough. AB - Acute cough due to viral upper respiratory tract infection (URI) is the most common form of cough and accounts for tremendous expenditure on prescription and non-prescription cough products worldwide. However, few agents have been shown in properly conducted clinical trials to be effective for cough due to URI. The present study evaluated the effect of benzonatate 200mg (B), guaifenesin 600 mg (G), their combination (B+G), and placebo (P) on capsaicin-induced cough in 30 adult nonsmokers with acute URI. On 3 separate days within a 7-day period, 1h after ingesting randomly assigned study drug in a double-blind fashion, subjects underwent capsaicin cough challenge testing, which involved inhalation of incremental doubling concentrations of capsaicin until the concentration of capsaicin inducing 5 or more coughs (C(5)) was attained. Each subject received 3 of 4 possible study drugs. G (p=0.01) but not B (p=NS) inhibited cough-reflex sensitivity (log C(5)) relative to P. The combination of B+G suppressed capsaicin induced cough to a greater degree than B alone (p<0.001) or G alone (p=0.008). The mechanism by which the combination of B+G causes a potentiation of antitussive effect remains to be elucidated. Our results suggest that B+G may be an effective therapy for acute cough due to the common cold (URI). PMID- 19121574 TI - Handling leachate from glass cullet stockpiles. AB - Mixed glass cullet (crushed recycled glass containers) is stockpiled uncovered before use as roadway construction aggregate or daily cover in landfills. Rainwater that leaches through the stockpiles dissolves and suspends contaminants such as those from food residuals and paper labels. The objective of this study was to determine leachate quantity and quality from cullet stockpiles as a basis for development of Best Management Practices (BMPs). Four 35-tonne field stockpiles were set up for leachate analysis and to determine the effects of mechanical turning treatment on the leachate. Field-collected leachate and laboratory-generated washwater of cullet (water:cullet=3:1 by weight) were both analyzed for basic wastewater parameters, which showed pollutant levels comparable to or higher than those of untreated domestic wastewater or urban stormwater. While organic contamination decreased substantially (e.g., washwater BOD>95% reduction), TKN and total-phosphorus levels in leachate ranged between 11.6-154mgL(-1) and 1.6-12.0mgL(-1), respectively, and remained comparable to levels found in untreated domestic wastewater after four months. Turning enhanced the degradation of the organic constituents inside the stockpiles, which was confirmed by elevated temperatures. Based on this study, leachate from glass cullet stockpiles should not be released to surface water. For leachate from long term cullet stockpiles, release to groundwater should be only done after treatment to reduce nitrogen levels. PMID- 19121575 TI - 19th century London dust-yards: a case study in closed-loop resource efficiency. AB - The material recovery methods used by dust-yards in early 19th century London, England and the conditions that led to their development, success and decline are reported. The overall system developed in response to the market value of constituents of municipal waste, and particularly the high coal ash content of household 'dust'. The emergence of lucrative markets for 'soil' and 'breeze' products encouraged dust-contractors to recover effectively 100% of the residual wastes remaining after readily saleable items and materials had been removed by the thriving informal sector. Contracting dust collection to the private sector allowed parishes to keep the streets relatively clean, without the need to develop institutional capacity, and for a period this also generated useful income. The dust-yard system is, therefore, an early example of organised, municipal-wide solid waste management, and also of public-private sector participation. The dust-yard system had been working successfully for more than 50 years before the Public Health Acts of 1848 and 1875, and was thus important in facilitating a relatively smooth transition to an institutionalised, municipally-run solid waste management system in England. The dust-yards can be seen as early precursors of modern materials recycling facilities (MRFs) and mechanical-biological treatment (MBT) plants; however, it must be emphasised that dust-yards operated without any of the environmental and occupational health considerations that are indispensable today. In addition, there are analogies between dust-yards and informal sector recycling systems currently operating in many developing countries. PMID- 19121576 TI - Solid waste treatment opportunities in the Palestinian authority areas. AB - Municipal services in the Palestinian Authority (PA) areas, including the West Bank and Gaza Strip (WBGS), are facing serious difficulties that have been intensified following the outbreak of the Palestinian uprising in late September 2000. The solid waste management services, being the most essential services provided by the municipalities and village councils, are mostly affected by the ongoing harsh situation and hence proper solutions that take into account the actual amount of generated municipal solid waste and its composition is a pre requisite for planning proper treatment. Hence, a study was carried out to identify the actual status of solid waste in eight West Bank districts. A social survey was also conducted to collect information concerning the level of public awareness among communities surveyed to the perception of solid waste recycling and reuse. The results of the survey conducted in 2001-2002 were later reviewed during July-October 2008 to assess if the trend of domestic solid waste generation had changed. Based on the survey and post-assessment, it is found that political and economic conditions have both significantly impacted the trend of generated municipal solid waste and since no improvements in either condition are forthcoming, it is concluded that survey results could be used in a planning study. A possible handling of the generated wastes may entail transferring the recyclable waste to Israeli recycling industries, and in constructing three composting plants in different accessible locations in the West Bank. PMID- 19121577 TI - Stat3 activation is required for the growth of U87 cell-derived tumours in mice. AB - Previously we reported that Stat3 is persistently activated in GBM tumours and derived cell lines. Hypoxia, necrosis and neo-angiogenesis are hallmarks of GBM. To unfold the contribution of activated Stat3 to the growth of GBM, we generated human GBM cell line (U87)-derived stable clones expressing a dominant negative mutant (DN)-Stat3 in a hypoxia-inducible manner, and examined their tumour forming potentials in immune-compromised mice. We found that the parental and vector control cell-derived tumours grew steadily, whereas DN-Stat3-expressing clone-derived tumours failed to grow beyond 2mm of thickness in mouse flanks. This blockade of tumour growth was associated with induction of tumour cell apoptosis and suppression of tumour angiogenesis. Consistent with this, mice bearing orthotopically implanted DN-Stat3-expressing clones survived significantly longer than the control mice. These data suggest that activated Stat3 is required for the growth of GBM, and that targeting Stat3 may intervene with the growth of GBM. PMID- 19121579 TI - Internal mammary sentinel nodes: ignore, irradiate or operate? AB - INTRODUCTION: This study describes the results of internal mammary chain (IMC) biopsy, identifying factors that predict 'hot spots' and nodal metastases for patients in whom mapped IMC nodes were routinely dissected. METHODS: The nodal basin and status of every axillary and IMC site identified by lymphoscintigraphy were examined. Binary logistic regression analysed the relationship of several patients and tumour factors with IMC hot spots and metastases. RESULTS: Ninety of 490 patients (18.4%) had IMC sentinel lymph nodes (SLNs) identified by lymphatic mapping and dissected, and 20 of these (22.2%) were found to have metastases. Mapping to the IMC was most likely for women aged under 35 years (29.4%) (p=0.117), women aged 35-44 (22.6%) (p=0.034) or those with medial (23.7%) or central tumour location (22.2%) (p=0.014; p=0.062, respectively). Predictors of IMC positivity included age <35 years (p=0.063), grade 3 histology (p=0.018) and lymphatic vascular invasion (LVI) (p=0.032). Although IMC positivity was more likely with positive axillary nodes, this trend was not significant. CONCLUSION: We identified several factors (age <35 years, tumour grade and LVI) that independently predict IMC SLN identification and positivity for patients with stage I or II breast cancer. Where IMC hot spots are not dissected, we predict IMC positivity of 50% or more for young women (<35 years) or women with high grade or LVI positive tumours, and these women may benefit from more intensive chemotherapy and radiotherapy to the IMC. PMID- 19121580 TI - Safety and pharmacokinetics of plasma-derived mannose-binding lectin (MBL) substitution in children with chemotherapy-induced neutropaenia. AB - Mannose-binding lectin (MBL)-deficient children with cancer may benefit from substitution of the innate immune protein MBL during chemotherapy-induced neutropaenia. We determined the safety and pharmacokinetics of MBL substitution in a phase II study in MBL-deficient children. Twelve MBL-deficient children with cancer (aged 0-12 years) received infusions of plasma-derived MBL once, or twice weekly during a chemotherapy-induced neutropaenic episode (range: 1-4 weeks). Four patients participated multiple times. Target levels of 1.0 microg/ml were considered therapeutic. In total, 65 MBL infusions were given. No MBL-related adverse reactions were observed, and the observed trough level was 1.06 microg/ml (range: 0.66-2.05 microg/ml). Pharmacokinetics were not related to age after correction for body weight. The half-life of MBL, for a child of 25 kg, was 36.4h (range: 23.7-66.6h). No anti-MBL antibodies were measured 4 weeks after each MBL course. Substitution therapy with MBL-SSI twice weekly was safe and resulted in trough levels considered protective. PMID- 19121581 TI - Statistical media optimization for growth and PHB production from methanol by a methylotrophic bacterium. AB - Media components were optimized by statistical design for cell growth and PHB production of Methylobacterium extorquens DSMZ 1340. Four important components of growth media were optimized by central composite design. The growth increased from an OD=1.35 for Choi medium as control to an OD=2.15 for optimal medium. Then media components for PHB production were optimized. Optimization of five important factors was conducted by response surface method. The optimal composition of PHB production medium was found to be at 7.8 (g/L) Na2HPO4 x 12H2O, and surprisingly at zero concentration of (NH4)2SO4, KH2PO4, MgSO4 and MnSO4. The PHB production was found to be 2.95 (g/L) at this medium. RSM results indicated that a deficiency of nitrogen and magnesium is crucial for PHB accumulation in this microorganism. Also, PHB production was carried out in a 5 L fermentor at the optimum condition which resulted in 9.5 g/L PHB and 15.4 g/L cell dry weight with 62.3% polymer content. PMID- 19121582 TI - Carbonic anhydrase inhibitors: inhibition studies of a coral secretory isoform with inorganic anions. AB - The inhibition of a coral carbonic anhydrase (CA, EC 4.2.1.1) has been investigated with a series of inorganic anions such as halogenides, pseudohalogenides, bicarbonate, carbonate, nitrate, nitrite, hydrogen sulfide, bisulfite, perchlorate, sulfate. The full-length scleractinian coral Stylophora pistillata CA, STPCA, has a significant catalytic activity for the physiological reaction of CO(2) hydration to bicarbonate, similarly to the ubiquitous human isoforms hCA I (cytosolic) and hCA VI (secreted). The best STPCA anion inhibitors were bromide, iodide, carbonate, and sulfamate, with inhibition constants of 9.0 10.0 microM. PMID- 19121578 TI - The cancer survival gap between elderly and middle-aged patients in Europe is widening. AB - The present study is aimed to compare survival and prognostic changes over time between elderly (70-84 years) and middle-aged cancer patients (55-69 years). We considered seven cancer sites (stomach, colon, breast, cervix and corpus uteri, ovary and prostate) and all cancers combined (but excluding prostate and non melanoma skin cancers). Five-year relative survival was estimated for cohorts of patients diagnosed in 1988-1999 in a pool of 51 European populations covered by cancer registries. Furthermore, we applied the period-analysis method to more recent incidence data from 32 cancer registries to provide 1- and 5-year relative survival estimates for the period of follow-up 2000-2002. A significant survival improvement was observed from 1988 to 1999 for all cancers combined and for every cancer site, except cervical cancer. However, survival increased at a slower rate in the elderly, so that the gap between younger and older patients widened, particularly for prostate cancer in men and for all considered cancers except cervical cancer in women. For breast and prostate cancers, the increasing gap was likely attributable to a larger use of, respectively, mammographic screening and PSA test in middle-aged with respect to the elderly. In the period analysis of the most recent data, relative survival was much higher in middle-aged patients than in the elderly. The differences were higher for breast and gynaecological cancers, and for prostate cancer. Most of this age gap was due to a very large difference in survival after the 1st year following the diagnosis. Differences were much smaller for conditional 5-year relative survival among patients who had already survived the first year. The increase of survival in elderly men is encouraging but the lesser improvement in women and, in particular, the widening gap for breast cancer suggest that many barriers still delay access to care and that enhanced prevention and clinical management remain major issues. PMID- 19121583 TI - Synthesis and biological evaluation of radioiodinated 2NUBTA as a cerebral ischemia marker. AB - N-[4-(Benzothiazol-2-yl)phenyl]-11-(2-nitroimidazole-1-yl)undecanamide (2NUBTA) was synthesized and radiolabeled with iodine-131. In vitro evaluation of the [(131)I]2NUBTA using murine sarcoma S180 cells showed increase in radioactivity in hypoxic cells up to 4h, while it was not in aerobic cells. Its potential as a cerebral ischemia marker was evaluated using gerbil stroke models that had been subjected to right common carotid artery ligation to produce cerebral ischemia. The uptake in the right cerebral hemisphere decreased slowly than that of the left and the right/left hemisphere uptake ratios increased with time going on. It indicated that [(131)I]2NUBTA might be a possible cerebral ischemia marker. PMID- 19121584 TI - Determination of 1-aryl-4-propylpiperazine pKa values: the substituent on aryl modulates basicity. AB - In order to design a potential drug, it is important to know its pK(a) because the protonation state of the molecule will be critical for ligand-receptor interaction and for the pharmacokinetic of the molecule. pK(a) values of a series of 1-(substitutedphenyl)-4-propylpiperazines were measured to study how the presence of a substituent on the phenyl ring modulates the basicity of N-4 nitrogen. pK(a) values indicated that the position of the substituent was crucial. In general, the introduction of the substituent in ortho-position of the phenyl ring increased the basicity of the molecule. This effect appeared to be related to steric and conformational effects and not to the electronic properties of the substituent. On the other hand, meta- and para-substituted derivatives showed a slight decrease of pK(a) that was qualitatively consistent with the electronic properties of the substituent. PMID- 19121585 TI - In vitro study on the interaction between thiophanate methyl and human serum albumin. AB - Thiophanate methyl (MT) is one of the widely used fungicides to control important fungal diseases of crops, which has led to potential toxicological risk to public health. Several different transport proteins exist in blood plasma, but albumin only is bound by a wide diversity of xenobiotics reversibly with high affinity. We studied the interaction of MT with human serum albumin by using spectroscopic methods including fluorescence quenching technology, UV and Fourier transform infrared (FT-IR) spectroscopy under simulative physiological conditions. The result of fluorescence titration revealed that MT could quench the intrinsic fluorescence of HSA. The binding process was exothermic and spontaneous, as indicated by the thermodynamic analyses. In addition, the studies of FT-IR spectroscopy showed that the binding of MT to HSA changed molecular conformation of HSA. The results obtained from molecular modeling showed that the interaction between MT and HSA was dominated by hydrophobic force, and there was also hydrogen bond interaction between the pesticide and the residues of HSA, which was in good agreement with the result of binding mode. PMID- 19121587 TI - Reliability difference between spectral and entropic measures of erector spinae muscle fatigability. AB - This study investigated back muscle fatigability of the erector spinae (ES) muscles during submaximal contractions with the use of surface electromyography (sEMG). It was important to confirm the reliability of measurements for characterizing neuromuscular alterations by establishing the difference between the Shannon (information) entropy and the power spectrum analysis in subjects with low back pain (LBP). The data was collected on two different days, one week apart, and the between-days reliability of these measures was examined. Thirty two gender-matched subjects completed the modified Sorenson test; 16 of the subjects were female while 16 were male. The entropy of the sEMG signal was more reliable than either the median frequency (MF) or the slope of the MF. The intraclass correlation coefficient (ICC) was higher for the entropy than for the MF slope. The ICC values of entropy for between-day measurements were higher (0.82-0.85) than MF (0.54-0.64) and MF slope (0.26-0.30). The standard errors of measurement (SEM) values for entropy were lower (0.04-0.05) than MF (3.10-3.60) and MF slope (0.03-0.04). The Pearson correlation coefficients of the entropy were significantly higher (0.75-0.77) than those of the MF (0.38-0.47) and the MF slope (0.15-0.18). Therefore, the results of this study indicated that the entropy analysis could provide a reliable measure of muscle fatigability. PMID- 19121588 TI - Meniscal tissue explants response depends on level of dynamic compressive strain. AB - OBJECTIVE: Following partial meniscectomy, the remaining meniscus is exposed to an altered loading environment. In vitro 20% dynamic compressive strains on meniscal tissue explants has been shown to lead to an increase in release of glycosaminoglycans from the tissue and increased expression of interleukin-1alpha (IL-1alpha). The goal of this study was to determine if compressive loading which induces endogenously expressed IL-1 results in downstream changes in gene expression of anabolic and catabolic molecules in meniscal tissue, such as MMP expression. METHOD: Relative changes in gene expression of MMP-1, MMP-3, MMP-9, MMP-13, A Disintegrin and Metalloproteinase with ThromboSpondin 4 (ADAMTS4), ADAMTS5, TNFalpha, TGFbeta, COX-2, Type I collagen (COL-1) and aggrecan and subsequent changes in the concentration of prostaglandin E(2) released by meniscal tissue in response to varying levels of dynamic compression (0%, 10%, and 20%) were measured. Porcine meniscal explants were dynamically compressed for 2h at 1Hz. RESULTS: 20% dynamic compressive strains upregulated MMP-1, MMP-3, MMP 13 and ADAMTS4 compared to no dynamic loading. Aggrecan, COX-2, and ADAMTS5 gene expression were upregulated under 10% strain compared to no dynamic loading while COL-1, TIMP-1, and TGFbeta gene expression were not dependent on the magnitude of loading. CONCLUSION: This data suggests that changes in mechanical loading of the knee joint meniscus from 10% to 20% dynamic strain can increase the catabolic activity of the meniscus. PMID- 19121589 TI - A randomised open-label trial comparing long-term sub-cutaneous low-molecular weight heparin compared with oral-anticoagulant therapy in the treatment of deep venous thrombosis. AB - OBJECTIVE: To evaluate whether low-molecular-weight heparin (LMWH) could be equally (or more) effective than oral anti-vitamin-K agents (AVK) in the long term treatment of deep venous thrombosis (DVT). DESIGN: A randomised, open-label trial. MATERIAL AND METHODS: In this trial, 241 patients with symptomatic proximal DVT of the lower limbs confirmed by duplex ultrasound scan were included. After initial LMWH, patients received 6 months of treatment with full therapeutic dosage of tinzaparin or acenocoumarol. The primary outcome was the 12 month incidence of symptomatic recurrent venous thrombo-embolism (VTE). Duplex scans were performed at 6 and 12 months. RESULTS: During the 12-month period, six patients (5%) of 119 who received LMWH and 13 (10.7%) of 122 who received AVK had recurrent VTE (p=0.11). In patients with cancer, recurrent VTE tended to be lower in the LMWH group (two of 36 [5.5%]) vs. seven of 33 [21.2%]; p=0.06). One major bleeding occurred in the LMWH group and three in the AVK group. Venous re canalisation increased significantly at 6 months (73.1% vs. 47.5%) and at 12 months (91.5% vs. 69.2%) in the LMWH group. CONCLUSIONS: Tinzaparin was more effective than AVK in achieving re-canalisation of leg thrombi. Long-term tinzaparin was at least as efficacious and safe as AVK for preventing recurrent VTE, especially in patients with cancer. PMID- 19121590 TI - Just world beliefs moderate the relationship of pain intensity and disability with psychological distress in chronic pain support group members. AB - The impact of pain beliefs on coping and adjustment is well established. However, less is known about how beliefs unrelated to pain might impact upon this experience. In particular, just world beliefs could impact upon and be influenced by chronic pain, given that pain is not experienced in a vacuum but instead is experienced in a social context where justice issues are potentially salient. The focus of this study was the ability of personal and general just world beliefs to moderate the relationships psychological distress held with pain intensity and disability in chronic pain. The sample (N=95) was recruited from members of arthritis and fibromyalgia support groups to investigate these social beliefs in a controlled community pain context. A cross-sectional, questionnaire design was adopted. The personal just world belief was endorsed significantly more than the general just world belief, and endorsement of the personal just world belief was negatively correlated with pain intensity, disability and psychological distress, while the general just world belief was unrelated to these variables. When interaction terms relating to personal and general just world beliefs were entered simultaneously into regression analyses, the personal just world belief did not predict psychological distress. However, pain intensity positively predicted psychological distress at low but not high levels of the general just world belief, while disability predicted psychological distress at low and high levels of this belief. This suggests that a strong general just world belief has implications for psychological well-being in chronic pain, and as such this belief may occupy a potential coping function in this context. PMID- 19121591 TI - Flow imaging of fluids in porous media by magnetization prepared centric-scan SPRITE. AB - MRI has considerable potential as a non-destructive probe of porous media, permitting rapid quantification of local fluid content and the possibility of local flow visualization and quantification. In this work we explore a general approach to flow velocity measurement in porous media by combining Cotts pulsed field gradient flow encoding with SPRITE MRI. This technique permits facile and accurate flow and dispersion coefficient mapping of fluids in porous media. This new approach has proven to be robust in characterizing fluid behavior. This method is illustrated through measurements of flow in pipes, flow in sand packs and flow in porous reservoir rocks. Spatially resolved flow maps and local fluid velocity distribution were acquired. PMID- 19121586 TI - miRNA regulation of cytokine genes. AB - In this review we discuss specific examples of regulation of cytokine genes and focus on a new mechanism involving post-transcriptional regulation via miRNAs. The post-transcriptional regulation of cytokine genes via the destabilizing activity of AU-rich elements [AREs] and miRNAs is a pre-requisite for regulating the half-life of many cytokines and achieving the temporal and spatial distributions required for regulation of these genes. PMID- 19121592 TI - High-resolution 1H MAS RFDR NMR of biological membranes. AB - The combination of magic angle spinning (MAS) with the high-resolution (1)H NOESY NMR experiment is an established method for measuring through-space (1)H...(1)H dipolar couplings in biological membranes. The segmental motion of the lipid acyl chains along with the overall rotational diffusion of the lipids provides sufficient motion to average the (1)H dipolar interaction to within the range where MAS can be effective. One drawback of the approach is the relatively long NOESY mixing times needed for relaxation processes to generate significant crosspeak intensity. In order to drive magnetization transfer more rapidly, we use solid-state radiofrequency driven dipolar recoupling (RFDR) pulses during the mixing time. We compare the (1)H MAS NOESY experiment with a (1)H MAS RFDR experiment on dimyristoylphosphocholine, a bilayer-forming lipid and show that the (1)H MAS RFDR experiment provides considerably faster magnetization exchange than the standard (1)H MAS NOESY experiment. We apply the method to model compounds containing basic and aromatic amino acids bound to membrane bilayers to illustrate the ability to locate the position of aromatic groups that have penetrated to below the level of the lipid headgroups. PMID- 19121593 TI - Simulation of DNMR spectra using propagator formalism and Monte Carlo method. AB - A new program-ProMoCS-is presented for the simulation of dynamic nuclear magnetic resonance spectra. Its algorithm is based on the Monte Carlo method as the one of the previously introduced MC-DNMR but the theory of ProMoCS is explained by using the statistical approach of propagator formalism. Our new program is suitable for the calculation of dynamic NMR spectra of spin systems up to 12 1/2 spin nuclei, several conformers and any type of exchange between them. Mutual exchange of coupled spins can be simulated as well. While it keeps the main advantage of the Monte Carlo based method: calculation with significantly smaller matrices as compared with programs based on the simulation of the average density matrix, the maximum number of nuclei is increased significantly. Thus spectra of such systems can be simulated that was impossible previously. PMID- 19121594 TI - Active compensation of rf-pulse transients. AB - A new approach to compensate rf-pulse transients is proposed. Based on the idea of the response theory of a linear system, a formula is derived to obtain the required excitation voltage profile back from the intended target rf-pulse shape. The validity of the formula is experimentally confirmed by monitoring the rf field created inside the sample coil with a pickup coil. Since this approach realizes accurate rf-pulse shapes without reducing the Q-factor of the tank circuit of the probe, it can be used not only to suppress the transient tail of the rf-pulse, but also as a general concept for accurate rf-pulsing. PMID- 19121595 TI - High-resolution retinal imaging in young children using a handheld scanner and Fourier-domain optical coherence tomography. AB - PURPOSE: To test the feasibility and applicability of a handheld probe for Fourier-domain optical coherence tomography (Fd-OCT) retinal imaging in infants and children. METHODS: Thirty children ages 7 months to 9.9 years, with (10 of 30) or without (20 of 30) retinal pathology, were imaged with Fd-OCT. Imaging was performed under sedation in 10 of 30 children ages 7 months to 3.7 years. A high resolution Fd-OCT system (axial resolution: 4.5 mum; acquisition speeds: 1000 A scans/frame, 9 frames/second), constructed at the UC Davis Medical Center, in conjunction with a handheld scanner, was used for retinal imaging. RESULTS: Useful images were obtained from all selected patients. Image acquisition was possible in a conscious state in children as young as 3 years of age. All children tolerated the tests well. The most challenging situation for young children was the lack of an internal fixation target and the moving scanning line, which usually distracted them from a steady fixation. Despite these problems, image quality was comparable with scans previously obtained from an adult population. CONCLUSIONS: The flexible handheld scanner in association with high acquisition speed and high-resolution Fd-OCT allows retinal imaging in infants and children. This technology provides high-resolution documentation of retinal structure in a pediatric population for the first time. PMID- 19121596 TI - Field testing of the plusoptiX S04 photoscreener. AB - PURPOSE: To evaluate the diagnostic accuracy of the plusoptiX S04 digital photoscreener in a school screening program. METHODS: Between 2006 and 2007, 1343 information pamphlets/consent forms were sent to all junior kindergarten students in a local school district. Assistants from a local public health unit photographed 307 children. Of these, 271 children received an independent ophthalmic examination by a physician. Photographic results were compared with the those of the ophthalmic examination. Amblyopia risk factors were defined as anisometropia >1 D (sphere or cylinder), astigmatism >1.25 D, myopia >3 D, hyperopia >3.5 D, any manifest strabismus, and any media opacity. RESULTS: Photographic and examination results agreed in 94% of cases. Sensitivity in detecting amblyopia risk factors was 83%; specificity was 95%. The positive and negative predictive values were 73% and 97%, respectively. The untestable/unusable rate was 1%. CONCLUSIONS: These results compare favorably with a previously reported (but no longer available) digital photoscreening camera and are superior to results obtained with other off-axis photoscreening devices that require human interpretation. On the basis of these results, in a real-world screening program, the camera would falsely refer 4% of those screened and would fail to correctly refer 2%. The accuracy of the plusoptiX S04 camera in detecting amblyopia risk factors appears sufficiently high to consider its further deployment in a widespread school screening program. PMID- 19121598 TI - Population genetics of six miniSTR loci (D1S1677, D2S441, D4S2364, D10S1248, D14S1434, D22S1045) in a Polish population. AB - Allele frequency data and forensic efficiency parameters for six miniSTR loci: D1S1677, D2S441, D4S2364 (NC01), D10S1248, D14S1434, D22S1045 (NC02) were estimated from a sample of 116 unrelated individuals from Poland. No significant deviations from Hardy-Weinberg equilibrium expectations were detected. The combined power of discrimination for the six studied loci was 0.999995383. PMID- 19121597 TI - Correcting for fat mass improves DXA quantification of quadriceps specific strength in obese adults aged 50-59 years. AB - Dual-energy X-ray absorptiometry (DXA) is widely used for bone mineral density and body composition assessments. However, DXA is known to overestimate muscle mass in obese adults. We used single-slice CT (ssCT) to derive a correction factor to enhance accuracy of DXA estimation of specific strength (strength per unit muscle). One hundred and sixty-two adults (age: 55.0+/-2.7 yr, range: 50-59) were enrolled in this cross-sectional study and divided into groups based on body mass index (BMI: <30, 30-35, and > or =35). BMI groups did not differ in age, knee extensor strength (KES), thigh lean mass by DXA, or quadriceps cross sectional area (CSA) by ssCT. Specific strength (KES/CSA) correlated with an uncorrected estimate of DXA-specific strength (r = 0.82, 0.53, 0.84 and 0.74, 0.59, 0.57, p < 0.001) in the lowest to highest BMI groups in men and women, respectively. Stronger correlations were achieved through correcting for BMI, age, and sex in estimating DXA-specific strength (r = 0.81, 0.79, and 0.96 in the lowest to highest BMI groups in men and 0.94, 0.81, 0.85 in women, p < 0.0001). Quantification of knee extensor-specific strength by DXA in men with BMI > 30 and all BMI groups in women greatly improved using a correction factor for DXA estimates of thigh lean mass. PMID- 19121599 TI - An accidental case of aconite poisoning due to Kampo herbal medicine ingestion. AB - An accidental case of aconite intoxication occurred after a patient took a therapeutic dose of Kampo herbal medicine containing Aconiti tuber, Uzu but had used the wrong decoction procedure. The poisoning was likely caused by an increased level of Aconitum alkaloids in the decoction; the patient developed aconite intoxication due to incomplete decoction. Aconitum alkaloid levels in the leftover solution which the patient had drunk and in the decoction extracted from 3g Uzu were determined. It was found that decoction makes the medicine safer to drink. Older individuals, especially those with dementia, have a higher risk of aconite poisoning because they sometimes do not boil the medicine appropriately. PMID- 19121600 TI - The role of environmental and individual factors in the social epidemiology of chikungunya disease on Mayotte Island. AB - This article examines the role of environmental and individual factors in the social epidemiology of chikungunya disease on the island of Mayotte (South western Indian Ocean). In an epidemic setting, an interdisciplinary study combining interviews, observations, and serological tests was conducted to: (1) estimate the frequency and social distribution of chikungunya disease and (2) identify its principal cognitive, behavioral, and environmental determinants within a stratified random sample of the Mayotte population (n=888). Semi parametric tests and multiple correspondence analyses were used to describe the statistical relationships between the different classes of variables examined in this study and the presence of antibodies attributable to chikungunya. These analyses highlighted differences between two main types of populations: one more autochthonous, more urban and better educated population, which shared 'legitimate' representations of the disease-from a biomedical viewpoint; and the other more migrant, more suburban, and more deprived, which is characterized by folk theories of chikungunya virus infection. Moreover, a series of logistic regression models revealed that social disparities in the distribution of virus infection were primarily structured by the housing conditions and cognitive representations of the disease held by the participants. These results suggest that environmental and individual factors are equally crucial in epidemic settings, and that they could explain, to a considerable extent, the social differences observed in morbidity associated with recent emerging infectious diseases in tropical countries. PMID- 19121601 TI - Prediction of outcome following hypoxia/ischaemia in the human infant using cerebral impedance. AB - OBJECTIVE: Changes in cerebral impedance in the newborn piglet are able to discriminate, within 1-2 h of acute hypoxia, between animals which will have a good neurological outcome, and those who have suffered more severe hypoxia resulting in poor outcome. The aim of this study was to determine if cerebral impedance could be used to identify those human infants with an encephalopathy following acute hypoxia who subsequently have a poor neurological outcome. It is these infants who may benefit most from neural rescue treatment. METHODS: Twenty four newborn term infants with evidence of severe acute intrapartum hypoxia and encephalopathy were studied. Bioimpedance spectroscopy was commenced as soon as possible after birth and repeated every 30 min until the infant was 12 h old. Neurodevelopmental outcome was assessed at 12 months of age. RESULTS: Although cerebral impedance was different to control values, there was no significant difference in cerebral impedance between hypoxic babies with normal and those with abnormal development. CONCLUSION: Cerebral impedance was increased in hypoxic babies, as predicted from animal data, but the method was not suitable for discrimination of outcome. SIGNIFICANCE: Cerebral impedance is not useful for early identification of infants who subsequently have a poor outcome after acute intrapartum hypoxia and who may benefit from neural rescue treatment. PMID- 19121602 TI - Increase of theta/gamma ratio is associated with memory impairment. AB - OBJECTIVE: In this study the theta/gamma ratio was investigated as early marker of cognitive decline. METHODS: Forty-nine subjects with mild cognitive impairment (MCI) underwent EEG recording and MRI scan. The theta/gamma ratio of the relative power at the peak frequency was computed. Based on the tertiles values of the ratio, three groups with increasing values of theta/gamma ratio were obtained. The groups were characterized by the performance on cognitive tests. Changes in functional brain connectivity, as expressed by interhemisperic and intrahemispheric EEG linear coherence in the groups were also evaluated. RESULTS: Increase in theta/gamma ratio was associated with impairment in memory tests. This relationship was confirmed by correlation and multiple regression analysis. An independent association was found between theta/gamma ratio and alpha3/alpha2 power ratio. Coherence analysis showed modifications of interhemispheric functional coupling on temporal regions on slow frequencies. CONCLUSIONS: Theta/gamma ratio of relative power at peak frequency is significantly associated to memory decline. It could be a useful tool in detecting MCI subjects which are at major risk to develop Alzheimer's disease (AD) or other dementias. SIGNIFICANCE: A global modulation of brain rhythms could be driven by the pathological alterations of theta/gamma ratio. PMID- 19121603 TI - Ultrastructure and molecular phylogeny of two heterolobosean amoebae, Euplaesiobystra hypersalinica gen. et sp. nov. and Tulamoeba peronaphora gen. et sp. nov., isolated from an extremely hypersaline habitat. AB - We isolated two amoebae, Tulamoeba peronaphora gen. et sp. nov. and Euplaesiobystra hypersalinica gen. et sp. nov. from the high salinity waters (293 300 per thousand salinity) of a Korean solar saltern. These new species show features typical of Heterolobosea - a limax form with eruptive pseudopodial formation, flattened/discoidal mitochondrial cristae, cysts with plugged pores, and no discrete, stacked dictyosomes. 18S rRNA gene phylogenies place both species within the Heterolobosea. Tulamoeba peronaphora appears to lack a flagellate phase, and has one cyst pore that penetrates the cyst wall. In 18S rRNA gene trees, Tulamoeba peronaphora is specifically related to Pleurostomum flabellatum, an extreme halophile that is observed only as a flagellate. Its next closest relatives are Naegleria and Willaertia. Euplaesiobystra hypersalinica has 2-4 cyst pores in the ectocyst wall (only), and has a bi-flagellated flagellate phase with no obvious cytostome. Its closest described relative is Heteramoeba clara, which is marine, has a cytostome, lacks cyst pores, and has a different nucleolus organization. The Euplaesiobystra hypersalinica 18S rRNA gene is 99.5% identical to a sequence accessed under the nomen nudum 'Plaesiobystra hypersalinica' - we consider them the same species. Tulamoeba peronaphora grows at 75-250 per thousand salinity, while E. hypersalinica grows at 100-300 per thousand (at least) salinity. Both amoebae seem to be 'extreme halophiles', and their ancestors invaded high salinity environments independently of each other. These results provide more evidence that there is a substantial ecological and phylogenetic diversity of heterotrophic eukaryotes capable of growing in very high salinity environments, and these ecosystems may be more complex than usually assumed. PMID- 19121605 TI - Cholera in Ethiopia in the 1990 s: epidemiologic patterns, clonal analysis, and antimicrobial resistance. AB - In 1993, after 6 years of absence, cholera re-emerged in the Horn of Africa. Following its introduction to Djibouti, the disease spread to the central and southern areas of Ethiopia reaching Somalia in 1994. Cholera outbreaks persisted in Ethiopia with a recrudescence of cases in 1998. Twenty-two Vibrio cholerae O1 strains, selected to represent the 1998 history of cholera in Ethiopia, were characterized by random amplified polymorphic DNA patterns, BglI ribotyping and antimicrobial susceptibility. All isolates showed a unique amplified DNA pattern and a prevalent ribotype B8a. All strains were multidrug-resistant and harboured an IncC plasmid which conferred resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole and trimethoprim. These findings indicate that a group of closely related V. cholerae O1 strains was responsible for the cholera epidemic in Ethiopia in 1998. PMID- 19121606 TI - Endoscopic injection of submucosal bulking agents for the management of incontinent catheterizable channels. AB - BACKGROUND: Urinary and fecal continence can be achieved by constructing catheterizable continent channels that provide access to the bladder and bowel. Some patients develop persistent stomal leakage. A minimally invasive method of injection with a bulking agent for treatment of stomal incontinence was evaluated. METHODS: A retrospective review identified patients with incontinence of their catheterizable continent urinary channel (CUC) and/or antegrade continence enema (ACE). All patients underwent circumferential endoscopic sub mucosal injection of the channel with a bulking agent, performed at the level of the continence mechanism. The type of injected material, number of procedures required, and success rates were evaluated. RESULTS: Out of 157 patients with a CUC and/or ACE (total of 164 stomas), eight patients underwent the minimally invasive therapy (total of nine stomas). The initial reconstructive procedure was appendicovesicostomy in one patient, ileovesicostomy (Monti) in seven patients, and ACE in two patients. Amount of bulking agent injected varied from 1.4 to 7 cc (mean 3.72 cc). Follow up ranged from 1 to 39 months (median 15 months). Two patients received multiple injections. One patient had injection of both a CUC and ACE. At the time of final follow up, 6/7 (86%) patients with a CUC and 1/2 (50%) with an ACE were continent per catheterizable channel. CONCLUSION: Injection of a bulking agent provides an excellent minimally invasive treatment alternative for incontinence of a catheterizable channel. PMID- 19121604 TI - Genetic stability of vaccine strain Salmonella Typhi Ty21a over 25 years. AB - The attenuated live bacterial vaccine strain Salmonella enterica Serovar Typhi Ty21a is the main constituent of Vivotif, the only licensed oral vaccine against typhoid fever. The strain was developed in the 1970s by chemical mutagenesis. In the course of this mutagenesis, a number of mutations were introduced into the vaccine strain. Characterisation of the vaccine strain during development as well as release of master- and working seed lots (MSL and WSL) and commercial batches is based on phenotypic assays assessing microbiological and biochemical characteristics of Ty21a. In the current study, we have analysed by DNA sequencing the specific mutations originally correlated with the attenuation of strain Ty21a. These data demonstrate the stability of these mutations for MSLs and WSLs of Ty21a produced between 1980 and 2005. Finally, we have confirmed the correlation of these genetic mutations with the expected phenotypic attenuations for the seed lots used in vaccine manufacture over 25 years. PMID- 19121607 TI - Analysis of in vitro lymphocyte proliferation as a screening tool for cellular immunodeficiency. AB - Measuring lymphocyte response to mitogens and antigens is a mainstay of screening for cellular immunodeficiency. Few reports analyze performance as a screening tool in diverse patient cohorts. We studied proliferation assays performed at Children's Hospital Boston from 1996 to 2003 using mitogens phytohemagglutinin (PHA), concanavalin A (CONA) and pokeweed mitogen, and antigens tetanus (TT) and diphtheria (DT) toxoids, and compared a subset of patients with T cell dysfunction with adult controls using receiver operating characteristic analysis. Results were correlated with clinical data. CONA was superior to PHA in identifying patients with immunodeficiency. TT was second best. Interpretation based on raw CPM, a stimulation index, or reference to simultaneous controls all performed equally. Combining data from multiple mitogens and/or antigens did not enhance performance. Proliferation testing is a useful component of screening for cellular immunodeficiency, but is not a sensitive predictor of cellular immune compromise or risk of opportunistic infection. PMID- 19121608 TI - Cardiac dysautonomia and arterial distensibility in essential hypertensives. AB - INTRODUCTION: The central nervous system plays an important role in the regulation of blood pressure: the sympathetic nervous system may be a primary contributor to the development of some forms of essential hypertension. Hypertension is also associated with reduced distensibility of large arteries. The aim of our study is the evaluation of a correlation between cardiac dysautonomia (evaluated by means of heart rate variability [HRV]) and altered artery distensibility (evaluated by means of measurement of the time interval from the onset of the QRS wave and the detection of the last Korotkoff sound [QKD interval]). MATERIALS AND METHODS: HRV and QKD interval were evaluated in 23 patients (60.9+/-8.7 years) with untreated hypertension and in 20 control subjects (53.2+/-16.8 years). QKD interval and QKD(100-60) (that is QKD for a 100 mm Hg systolic blood pressure and 60 bpm heart rate) were measured during a 24 hours blood pressure monitoring. HRV was evaluated by means of a spectral method. Three main spectral components were distinguished: very low frequency (VLF), low frequency (LF) and high frequency (HF) component. RESULTS: Patients with reduced QKD(100-60) interval show reduced total power and spectral components values, with higher LF/HF ratio in basal conditions in comparison with control group. In patients with hypertension, QKD(100-60) values correlated significantly with LF/HF ratio (Spearman r=-0.551; p=0.006), HF spectral component (Spearman r=0.630; p=0.001) and total power (Spearman r=0.426; p=0.004). CONCLUSIONS: Our results suggest that sympathetic overactivity may be the contributor of reduced arterial distensibility observed in patients with essential hypertension. PMID- 19121609 TI - Influence of setting liquid composition and liquid-to-powder ratio on properties of a Mg-substituted calcium phosphate cement. AB - The influence of four variables on various properties of a Mg-substituted calcium phosphate cement (CPC) was investigated. The variables were the heat treatment temperature of the precipitated powders, the composition of the setting liquid, the liquid-to-powder ratio (LPR), and the time over which hardened specimens were cured in air. The properties analysed were the phase composition of the starting powder, the initial setting time, the evolution of the storage shear modulus (G') and the loss shear modulus (G'') with the cement paste curing time (t), and the compressive strength. The presence of alpha-TCP in CPC facilitated the setting and hardening properties due to its progressive dissolution and the formation of brushite crystals. As far as the liquid composition is concerned, in cases where citric acid was used, adding a rheology modifier (10 wt.% polyethylene glycol or 0.5 wt.% hydroxyl propylmethylcellulose) to the acid led to an increase in the initial setting time, while an increase in the acid concentration led to a decrease in the initial setting time. The initial setting time showed to be very sensitive towards the LPR. The evolution of G' and G'' with curing time reflected the internal structural changes of cement pastes during the setting process. The compressive strength of the wet-hardened cement specimens with and without Mg increased with curing time increasing, being slightly higher in the case of Mg substituted CPC. The results suggest that Mg-substituted CPC holds a promise for uses in orthopaedic and trauma surgery such as for filling bone defects. PMID- 19121610 TI - In vivo and in vitro evaluation of flexible, cottonwool-like nanocomposites as bone substitute material for complex defects. AB - The easy clinical handling and applicability of biomaterials has become a focus of materials research due to rapidly increasing time and cost pressures in the public health sector. The present study assesses the in vitro and in vivo performance of a flexible, mouldable, cottonwool-like nanocomposite based on poly(lactide-co-glycolide) and amorphous tricalcium phosphate nanoparticles (PLGA/TCP 60:40). Immersion in simulated body fluid showed exceptional in vitro bioactivity for TCP-containing fibres (mass gain: 18%, 2 days, HAp deposition). Bone regeneration was quantitatively investigated by creating four circular non critical-size calvarial defects in New Zealand White rabbits. The defects were filled with the easy applicable cottonwool-like PLGA/TCP fibres or PLGA alone. Porous bovine-derived mineral (Bio-Oss) was used as a positive control and cavities left empty served as a negative control. The area fraction of newly formed bone (4 weeks implantation) was significantly increased for TCP-containing fibres compared to pure PLGA (histological and micro-computed tomographic analysis). A spongiosa-like structure of the newly formed bone tissue was observed for PLGA/TCP nanocomposites, whereas Bio-Oss-treated defects afforded a solid cortical bone. PMID- 19121611 TI - Breast cancer diagnosis: the role of stereotactic vacuum-assisted aspiration biopsy. AB - AIM: The aim of this study is to evaluate the diagnostic accuracy and impact of the stereotactic vacuum-assisted aspiration biopsy (VAB) as a surgical treatment for non-palpable breast lesions. METHODS: A retrospective analysis of the diagnostic and therapeutic management of lesions having undergone VAB treatment was conducted. From February 2003 to September 2007, 525 stereotactic VABs were performed on 504 women using an 11-gauge needle device. Of these, 201 lesions were treated surgically. The concordance between VAB results and final pathology report after surgical excision was evaluated. Also examined was the impact of VAB on the quality of the surgical treatment. RESULTS: Stereotactic VABs performed with an 11-gauge device showed an underestimation rate of 23.8% for atypical ductal or lobular hyperplasia (AH). For ductal carcinoma in situ (DCIS) the underestimation rate was 31% and the underestimation rate for lobular carcinoma in situ (LCIS) was 14%. Only 38.2% of the patients with non-palpable lesions (201/525) were treated surgically and only 4% (20/504) of the patients underwent more than one surgical intervention. The VAB underestimation caused mistakes in the planning of the surgical therapy in only 9 out of 201 interventions (4.4%). CONCLUSIONS: This study confirms the efficacy of the VAB procedure in the diagnosis of non-palpable breast lesions and demonstrates its usefulness in therapeutic surgical planning. VAB treatment allows for the reduction of the number of surgical procedures required to diagnose and treat non-palpable breast lesions. PMID- 19121612 TI - The use of free vascularised bone grafts in spinal reconstruction. AB - The use of free vascularised bone grafts (FVBGs) is an infrequently performed surgical technique for the reconstruction of spinal defects. This field of surgery brings many challenges concerning choice of FVBG, planning of the operative procedure and selection of recipient vessels. This study aims to report our experience with FVBGs, with special emphasis on the planning and surgical technique. Over a period of 10 years (1994-2004), we used FVBG for anterior spinal reconstruction in 23 patients. In 21 patients, a free vascularised fibular graft was used, and in two cases a free vascularised iliac crest graft was used. The spinal segments reconstructed involved the cervical spine (4 cases), the thoracic spine (13 cases) and the thoraco-lumbar and lumbo-sacral spine (6 cases). Re-vascularisation of the FVBG proved to be technically feasible in 22 patients, but failed in one fibular graft due to difficulties with recipient vessels in the lumbar spine. When necessary, the fibula was osteotomized and folded in a double-, triple- or quadruple-barrel construction to increase the strength of the reconstruction. Technical challenges were met with respect to the choice of the recipient vessel at various anatomical sites. The use of FVBG is a valuable technique for the reconstruction of complex spinal disorders. Successful execution requires microvascular expertise with respect to graft harvesting and appropriate choice of recipient vessels. Adequate preoperative planning in a multidisciplinary setting and adherence to the basic principles for spinal reconstruction are required. PMID- 19121613 TI - A comparative study on two kinds of surgical procedures of penile corpora cavernosa augmentation. AB - OBJECTIVE: Penile augmentation surgery is still a controversial issue because of the uncertain indication, the possibility of severe complications and a variety of surgical techniques. The purpose of this study is to provide two surgical procedures of penile corpora cavernosa augmentation and to investigate its effect by implanting autogenous saphenous vein grafts or expanded polytetrafluoroethylene (ePTFE) vessel patches. METHODS: Between January 2001 and December 2005, 20 patients underwent surgeries in which bilateral longitudinal incisions were placed on the tunica albuginea and the penile corpora cavernosa were extended by means of implantation of saphenous grafts or PTFE artificial vessel patches. The patients included in this study presented either with congenital idiopathic micropenis or normal penile length and perimeter (dysmorphophobia). Before the operation, the penile length and perimeter in the flaccid and erectile states were as follows: flaccid length 2.5-7.5 cm and flaccid perimeter 3.0-7.5 cm; erectile length 4.9-10.5 cm and erectile perimeter 4.5-10.0 cm. RESULTS: Immediately after surgery, the penile corpus circumferential measurements (on table), showed remarkable increases which were 1.0-2.3 cm and 1.5-3.0 cm in the flaccid and erectile states, respectively; then, at 12 months to 5 years' follow-up, these girth gains had reduced by 0.5-1 cm in some cases. All cases in the two groups obtained satisfactory surgical results with satisfactory erection and no serious complications, such as infection and fistula. In 20 cases, 17 married cases resumed regular and satisfactory sexual activities 1 month after the operation without any functional limitation. CONCLUSION: The two kinds of surgical procedures for augmenting penile corpora cavernosa were proved to be effective and reliable, with few complications. Both saphenous grafts and ePTFE artificial vessel patches are excellent materials for reconstructing the tunica albuginea. These augmenting phalloplasties can not only be used for patients with micropenis, but also applied to satisfy the cosmetic and functional requests of patients with normal penile length and perimeter. However, the long-term outcomes of these surgical procedures need a further, detailed follow-up study. PMID- 19121614 TI - The radiation exposure of Computed Tomographic Angiography (CTA) in DIEP flap planning: low dose but high impact. PMID- 19121615 TI - Deep dopamine extravasation injury: a case report. AB - We report the case of a 3-month-old girl with Down's syndrome, who sustained a deep and massive extravasation of dopamine, resulting in segmented, full thickness skin necrosis and transient brachial plexus palsy of her left upper extremity. The patient was managed conservatively, including wound care, de bridement of necrotic tissue, secondary wound healing and intensive physical therapy. The patient showed a satisfactory outcome with complete secondary closure of her wounds and full brachial plexus recovery after 1 year of follow up. The mechanism of action of dopamine in the deep soft tissue, the difficulties of an adequate diagnosis of a deep dopamine extravasation and alternative treatments are presented in this article. PMID- 19121616 TI - Haemodynamic changes in the fingers after free radial forearm flap transfer: a prospective study using SPP. AB - Harvesting the radial forearm flap may cause circulatory problems in the donor arm. To investigate the influence on donor hands after radial forearm flap harvesting, we assessed the process of circulatory changes prospectively by measuring skin perfusion pressure (SPP) that is clinically useful in detecting vascular lesions. The records of 17 patients (14 men and 3 women aged 59.7+/-11.8 years) who had undergone free radial forearm flap transfer for head and neck reconstruction, between December 2005 and April 2007, were analysed. SPP in the thumb (finger I), the middle finger (III) and the little finger (V) was measured in the 17 patients preoperatively and 1 month and 3, 6, 9 and 12 months postoperatively. All statistical tests were two sided, with a significance level defined as p<0.05. Preoperatively, baseline SPP was more dominant in finger I than in finger V. Postoperatively, SPP changed significantly in both fingers, while it showed no change in finger III and tended to be higher in finger I than in the other two. Harvesting the free radial forearm flap reduces skin perfusion in the fingers of the donor arm and, we assume, leads to a re-distribution of blood flow to the fingers, with the residual ulnar artery still supplying more blood flow to finger I than to finger V. This suggests the presence of an autoregulating mechanism whereby blood perfusion to the fingers is controlled by the physiological demands of individual fingers. PMID- 19121617 TI - Reconstruction of a red lip that has a defect in one half, using the remaining red lip. AB - It is difficult to reconstruct the red lip and achieve good functional results such as normal sphincter force, sensitivity and movement of the lip. In addition, it is also difficult to attain colour and texture matches. We reconstruct a red lip that has a defect in one half, using the remaining red lip. Using the technique reported herein, we can achieve excellent functional and aesthetic results. Functionally, sphincter force and sensation recover early and the movement of the lip returns to normal. In addition, the reconstructed lip develops a symmetrical and natural appearance. PMID- 19121618 TI - Stripping custom microRNA microarrays and the lessons learned about probe-slide interactions. AB - Microarrays have been used extensively in gene expression profiling and genotyping studies. To reduce the high cost and enhance the consistency of microarray experiments, it is often desirable to strip and reuse microarray slides. Our genome-wide analysis of microRNA expression involves the hybridization of fluorescently labeled nucleic acids to custom-made, spotted DNA microarrays based on GAPSII-coated slides. We describe here a simple and effective method to regenerate such custom microarrays that uses a very low-salt buffer to remove labeled nucleic acids from microarrays. Slides can be stripped and reused multiple times without significantly compromising data quality. Moreover, our analyses of the performance of regenerated slides identifies parameters that influence the attachment of oligonucleotide probes to GAPSII slides, shedding light on the interactions between DNA and the microarray surface and suggesting ways in which to improve the design of oligonucleotide probes. PMID- 19121619 TI - Sample degradation leads to false-positive copy number variation calls in multiplex real-time polymerase chain reaction assays. AB - The recent implication of genomic copy number variations (CNVs) in multiple human genetic disorders has led to increased interest in CNV discovery technologies. There is a growing consensus that, in addition to the method used for detection, at least one additional technology should be employed for validation. Real-time quantitative polymerase chain reaction (qPCR) analysis, incorporating a normal (2N) copy number standard, is commonly used as a means of validating CNVs. Whereas it has previously been reported that formalin-fixed paraffin-embedded (FFPE) DNA samples can yield spurious CNV calls in real-time qPCR assays, here we report that sample degradation under standard laboratory storage conditions generates a significant increase in false-positive CNV results. Results suggest the possibility of biased degradation among genomic regions and emphasize the need to assess sample integrity immediately prior to real-time qPCR experiments. PMID- 19121620 TI - Ex vivo expansion of hematopoietic stem cell by fusion protein TAT-Zfx. AB - The relative inability of hemopoietic stem cells (HSCs) to reproduce themselves (self-renew) ex vivo imposes substantial limitations on the current use of HSC transplantation. Recently, the transcription factor Zfx has been demonstrated that played an important in controlling the self-renewal of hematopoietic stem cells. Here, we reported that Zfx could enable high-level expansion of HSCs in vitro, by combination of protein transduction domain, TAT. Furthermore, we also demonstrated that expanded HSCs population retains their normal in vivo potential of pluripotency. It is thus that TAT-Zfx has the potential to expand HSCs significantly in vitro, and will have enormous clinical potentials. PMID- 19121621 TI - A simple Calpha-SC potential with higher accuracy for protein fold recognition. AB - In this paper, an improved C(alpha)-SC energy potential designed for protein fold recognition was reported. It consists of three extremely simple interaction terms which are supposed to be the dominant interactions in protein folding: residue residue contact, hydrophobicity and pseudodihedral potentials. The potential function only contains 210 contacts, one hydrophobic and one torsion parameters, which have been optimized using an interior point algorithm of linear programming. Tests of the derived potential function on commonly used decoy sets illustrate that it outperforms most of the existing coarse-grained potentials in terms of its capabilities in recognizing native structures and consistency in achieving high Z-scores across decoy sets, and it has almost equivalent performance to the potentials which considered complex intra-molecular interactions. The results show that our scoring function is a generally prospective potential for protein structure prediction and modeling with regard to its recognition and computation efficacy. PMID- 19121622 TI - Bacterial neuraminidase increases IL-8 production in lung epithelial cells via NF kappaB-dependent pathway. AB - Bacterial neuraminidase, a sialic acid-degrading enzyme, is one of the virulent factors produced in pathogenic bacteria like as other bacterial components. However little is known about whether bacterial neuraminidase can initiate or modify a cellular response, such as cytokine production, in epithelial cells at infection and inflammation. We demonstrate here that bacterial neuraminidase, but not heat-inactivated neuraminidase, up-regulates expression of interleukin-8 (IL 8) mRNA and protein in lung epithelial A549 and NCI-H292 cells. We also show that bacterial neuraminidase significantly up-regulates IL-8 promoter activity as well as nuclear factor-kappaB (NF-kappaB) reporter activity. Moreover, inhibition of NF-kappaB signaling suppressed IL-8 mRNA expression induced by bacterial neuraminidase. Taken together, desialylation-induced IL-8 production in lung epithelial cells may play an important role in infection-associated inflammatory events. PMID- 19121623 TI - HDAC3 influences phosphorylation of STAT3 at serine 727 by interacting with PP2A. AB - Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors, and has been reported to be involved in the pathogenesis of various human diseases. Here, we show that treatment of HeLa cells with a histone deacetylase (HDAC) inhibitor, trichostatin A, or small-interfering RNA (siRNA) mediated repression of HDAC3, enhances phosphorylation of STAT3 at Ser727. Furthermore, dephosphorylation of STAT3 at Ser727 by protein phosphatase 2A (PP2A) was restored by treatment of cells with HDAC3 siRNA. We further found that formation of a complex between STAT3 and PP2A was enhanced in the presence of HDAC3. Importantly, small-interfering RNA-mediated repression of both HDAC3 and PP2A effectively enhanced leukemia inhibitory factor (LIF)-induced STAT3 activation. These results indicate that HDAC3 may act as a scaffold protein for PP2A to regulate the LIF/STAT3-mediated signaling pathway. PMID- 19121624 TI - Mechanism of inhibition of ribonucleotide reductase with motexafin gadolinium (MGd). AB - Motexafin gadolinium (MGd) is an expanded porphyrin anticancer agent which selectively targets tumor cells and works as a radiation enhancer, with promising results in clinical trials. Its mechanism of action is oxidation of intracellular reducing molecules and acting as a direct inhibitor of mammalian ribonucleotide reductase (RNR). This paper focuses on the mechanism of inhibition of RNR by MGd. Our experimental data present at least two pathways for inhibition of RNR; one precluding subunits oligomerization and the other direct inhibition of the large catalytic subunit of the enzyme. Co-localization of MGd and RNR in the cytoplasm particularly in the S-phase may account for its inhibitory properties. These data can elucidate an important effect of MGd on the cancer cells with overproduction of RNR and its efficacy as an anticancer agent and not only as a general radiosensitizer. PMID- 19121625 TI - Curcumin enhances non-inflammatory phagocytic activity of RAW264.7 cells. AB - Present study was performed to assess the effect of curcumin treatment on macrophage functions using RAW264.7 cells, a murine macrophage cell line. Phagocytic activity of RAW264.7 cells was enhanced by the treatment with curcumin for 48 hours while the nitric oxide synthesis from RAW264.7 cells following lipopolysaccharide exposure was blocked. The incubation of RAW264.7 cells with curcumin dose-dependently inhibited the stimulatory responses of macrophage triggered by lipopolysaccharide; the enhanced secretion of inflammatory cytokines such as TNF-alpha and IL-1beta and the up-regulated expression of surface antigens like CD14 and CD40. Curcumin alone, however, was able to increase the basal level of TNF-alpha secretion and elevated markedly the expression of CD14 and slightly CD40. The marked enhancement of both phagocytic activity and CD14 was detectable as early as 75min after curcumin treatment which is the minimum time period required for the phagocytosis and CD14 measurement, suggesting a signaling pathway distinct from that triggered by apoptotic cells. In conclusion, this study elucidates that curcumin treatment enhances the phagocytic activity with blocking nitric oxide synthesis, a scavenger function of macrophages in non inflammatory condition. In addition, this enhancement of phagocytic activity is triggered directly by the signals from curcumin itself not by apoptotic cells. PMID- 19121626 TI - CK2beta interacts with and regulates p21-activated kinases in Drosophila. AB - The role of CK2beta has been defined as the regulatory subunit of protein kinase CK2, which is a heterotetrameric complex composed of two CK2beta and two catalytic active CK2alpha subunits. The identification of other serine/threonine kinases such as A-Raf, Chk1, and c-Mos that interact with and are regulated by CK2beta has challenged this view and provided evidence for functions of CK2beta outside the CK2 holoenzyme. In this report we describe the first interaction of Drosophila CK2beta outside the CK2 holoenzyme with p21-activated kinase (PAK) proteins. This interaction is seen for distinct PAK and CK2beta isoforms. In contrast to the CK2alpha-CK2beta interaction, dimer formation of the CK2beta subunits is not a prerequisite for binding of PAK proteins. Our results support the idea that CK2beta can bind to PAK proteins in a CK2alpha independent manner and negatively regulates PAK kinase activity. PMID- 19121627 TI - Improvement of adaptive immunity by antigen-carrying biodegradable nanoparticles. AB - One of the most important aspects in vaccine development is to induce potent antigen-specific immune responses. In this study, we examined the immunological activities of antigen-carrying biodegradable poly(gamma-glutamic acid) (gamma PGA) nanoparticles (NPs) in mice. The immunization with ovalbumin (OVA)-carrying gamma-PGA NPs (OVA-NPs) could induce significant expansion of antigen-specific CD8(+) T cells. Unlike complete Freund's adjuvant, subcutaneous (s.c.) inoculation of OVA-NPs to footpad did not generate injection site swelling. Although OVA-NPs could induce both antigen-specific cellular and humoral immune responses, the dominant induction of either cellular or humoral immunity was found to depend on their administration routes. Strong antibody production was observed by s.c. immunization, yet no antibody was identified by intranasal immunization. Thus, gamma-PGA NPs are a safe and efficient antigen carrier with unique immunological properties. PMID- 19121628 TI - ROS and NF-kappaB are involved in upregulation of IL-8 in A549 cells exposed to multi-walled carbon nanotubes. AB - Carbon nanotubes (CNTs) have potential applications in biosensors, tissue engineering, and biomedical devices because of their unique physico-chemical, electronic and mechanical properties. However, there is limited literature data available concerning the biological properties and toxicity of CNTs. This study aimed to assess the toxicity exhibited by multi-walled CNTs (MWCNTs) and to elucidate possible molecular mechanisms underlying the biological effects of MWCNTs in A549 cells. Exposing A549 cells to MWCNTs led to cell death, changes in cell size and complexity, reactive oxygen species (ROS) production, interleukin-8 (IL-8) gene expression and nuclear factor (NF)-kappaB activation. Treatment of A549 cells with antioxidants prior to adding MWCNTs decreased ROS production and abrogated expression of IL-8 mRNA. Pretreatment of A549 cells with NF-kappaB inhibitors suppressed MWCNTs-induced IL-8 mRNA expression. These results indicate that MWCNTs are able to induce expression of IL-8 in A549 cells, at least in part, mediated by oxidative stress and NF-kappaB activation. PMID- 19121629 TI - Exercise training decreases expression of inflammation-related adipokines through reduction of oxidative stress in rat white adipose tissue. AB - Increased oxidative stress in adipocytes causes dysregulated expression of inflammation-related adipokines. We have examined the effects of exercise training on oxidative stress in rat white adipose tissue (WAT), especially focusing on inflammation-related adipokines. The levels of lipid peroxidation in WAT of exercise-trained (TR) rats were lower than those in control (C) rats. The content of manganese-containing superoxide dismutase in WAT of TR rats was increased as compared with those in C rats. In contrast, the expression of the NADPH oxidase NOX2 protein in WAT was downregulated by exercise training. Moreover, the levels of inflammation-related adipokines, such as tumor necrosis factor-alpha and monocyte chemoattractant protein-1, in WAT of TR rats were lower than those in C rats. The effects of exercise training were more remarkable in visceral WAT than in subcutaneous. These results suggest that exercise training decreases the expression of inflammation-related adipokines by reducing oxidative stress in WAT. PMID- 19121630 TI - Effects of folic acid deficiency and MTHFRC677T polymorphisms on cytotoxicity in human peripheral blood lymphocytes. AB - Apoptosis (APO) and necrosis (NEC) are two different types of cell death occurring in response to cellular stress factors. Cells with DNA damage may undergo APO or NEC. Folate is an essential micronutrient associated with DNA synthesis, repair and methylation. Methylenetetrahydrofolate reductase (MTHFR) regulates intracellular folate metabolism. Folate deficiency and MTHFR C677T polymorphisms have been shown to be related to DNA damage. To verify the cytotoxic effects of folate deficiency on cells with different MTHFR C677T genotypes, 15 human peripheral lymphocyte cases with different MTHFR C677T genotypes were cultured in folic acid (FA)-deficient and -sufficient media for 9 days. Cytotoxicity was quantified using the frequencies of APO and NEC as endpoints, the nuclear division index (NDI), and the number of viable cells (NVC). These results showed that FA is an important factor in reducing cytotoxicity and increasing cell proliferation. Lymphocytes with the TT genotype proliferated easily under stress and exhibited different responses to FA deficiency than lymphocytes with the CC and CT genotypes. A TT individual may accumulate more cytotoxicity under cytotoxic stress, suggesting that the effects of FA deficiency on cytotoxicity are greater than the effects in individuals with the other MTHFR C677T variants. PMID- 19121631 TI - Effect of the ghrelin receptor agonist TZP-101 on colonic transit in a rat model of postoperative ileus. AB - Ghrelin, the natural ligand of the growth hormone secretagogue receptor (ghrelin receptor), is an orexigenic gut hormone with prokinetic action in the upper gastrointestinal tract. Previously we have shown in a rodent model of postoperative ileus that the synthetic ghrelin receptor agonist TZP-101 prevents the delay in gastric emptying and improves small intestinal transit. The goal of the present study was to investigate whether TZP-101 affects colonic transit and food intake in rats with postoperative ileus. Fasted rats were treated with morphine and subjected to laparotomy under isoflurane anesthesia. Following surgery the animals were placed in clean home cages and fecal pellet output and food intake were monitored for 48 h. TZP-101 or vehicle were administered as 3 i.v. bolus infusions at 0 h, 2 h and 4 h post-surgery. TZP-101 (0.03-1 mg/kg) dose-dependently decreased the time to first bowel movement and increased fecal pellet output measured at 12 h and 24 h post-surgery compared to the vehicle. The administration of TZP-101 was not associated with a significant alteration in food intake. In conclusion, this study provides the first experimental evidence that a novel ghrelin receptor agonist improves large bowel function in rats with postoperative ileus, suggesting that TZP-101 may be useful in the clinic to accelerate upper gastrointestinal transit and to shorten the time to the first bowel movement following surgery. PMID- 19121632 TI - Stabilizing effects of eicosapentaenoic acid on Kv1.5 channel protein expressed in mammalian cells. AB - We investigated the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the stability of Kv1.5 channel protein. The expression and function of Kv1.5 (Kv1.5-FLAG) in transfected African green monkey kidney fibroblast cells as well as rat atrium were estimated by immunoblotting, immunoprecipitation, immunofluorescence and patch-clamp techniques. Both EPA and DHA immediately blocked Kv1.5 channel current in a dose-dependent manner, accompanied by reduction of their phosphorylation. Chronic treatment (for 12 h) with EPA at lower concentrations (0.3-10 muM) increased the level of Kv1.5-FLAG protein as well as Kv1.5 channel current without changes in its gating kinetics, prolonging its half-life; in contrast, both EPA and DHA at higher concentrations (30-100 muM) decreased the expression of Kv1.5-FLAG. EPA at the higher concentrations also decreased mRNA of Kv1.5 and synapse-associated protein 97 expression. EPA at the lower concentrations increased Kv1.5 expression in the endoplasmic reticulum, Golgi apparatus and cell membrane. EPA-induced increase of Kv1.5 channel expression and current was abolished by pretreatment with the protein transport inhibitor brefeldin A or colchicines, and by the Kv1.5 channel blocker 4 aminopyridine. Oral administration of EPA (30 mg/kg) increased the level of endogenous Kv1.5 in rat atria. These results indicate that chronic treatment with EPA at lower concentrations stabilizes Kv1.5 channel protein in the endoplasmic reticulum and Golgi apparatus thereby enhancing the Kv1.5 channel current on the cell membrane. PMID- 19121634 TI - Regulation of pancreas plasticity and malignant transformation by Akt signaling. AB - BACKGROUND & AIMS: Extensive evidence suggests that Akt signaling plays an important role in beta-cell mass and function, although its function in the regulation of the different pancreatic fates has not been adequately investigated. The goal of these studies was to assess the role of Akt signaling in the pancreatic differentiation programs. METHODS: For these experiments, we have generated a double reporter mouse model that provides activation of Akt signaling in a cell type-specific manner. This mouse model conditionally overexpresses a constitutively active form of Akt upon Cre-mediated recombination. Activation of Akt signaling in pancreatic progenitors and acinar and beta-cells was achieved by crossing this animal model to specific Cre-lines. RESULTS: We showed that overexpression of a constitutively active Akt in pancreatic and duodenal homeobox 1 (Pdx1) progenitors induced expansion of ductal structures expressing progenitor markers. This expansion resulted in part from increased proliferation of the ductal epithelium. Lineage-tracing experiments in mice with activation of Akt signaling in mature acinar and beta-cells suggested that acinar-to-ductal and beta-cell-to-acinar/ductal transdifferentiation also contributed to the expansion of the ductal compartment. In addition to the changes in cell plasticity, these studies demonstrated that chronic activation of Akt signaling in Pdx1 progenitors induced the development of premalignant lesions and malignant transformation in old mice. CONCLUSIONS: The current work unravels some of the molecular mechanisms of cellular plasticity and reprogramming, and demonstrates for the first time that activation of Akt signaling regulates the fate of differentiated pancreatic cells in vivo. PMID- 19121636 TI - Computational study of non-homogeneous distribution of Ca(2+) handling systems in cerebellar granule cells. AB - The spatiotemporal distribution of cytosolic free calcium concentration ([Ca(2+)](i)) in cerebellar granule cells (GrCs) is thought to be critical in defining the occurrence and direction of long-term changes in synaptic strength at cerebellar mossy fiber-GrC synapses. Despite this, the mechanisms responsible for shaping Ca(2+) transients in GrCs are not well understood. To investigate the interplay between Ca(2+) entry, extrusion, buffering and dendritic morphology in shaping Ca(2+) elevations in GrCs, we developed a model of Ca(2+) regulation in these cells and examined the requirements for reproducing fluorescence responses to depolarization and synaptic stimulation previously described in the literature. Two conclusions can be drawn from our simulation results. First, a significant progressive decrease in the amplitudes of depolarization-evoked fluorescence transients from the dendritic endings (digits) toward the soma of GrCs, can be reproduced in the model only if the density of Ca(2+) channels is considerably higher or the concentration of endogenous buffers is much lower in the digits than in the parent dendrites. In contrast, heterogeneities in the distribution of Ca(2+) pumps or in cytosolic fractional volume cannot account for the formation of [Ca(2+)](i) gradients in GrCs. Second, much lower amplitudes of fluorescence transients induced by depolarization and synaptic stimulation than expected from typical measurements of Ca(2+) and NMDA receptor-mediated currents can be reconciled with a pronounced slowing of the decay of fluorescence responses in the digits of GrCs after introducing a high-affinity Ca(2+) indicator if a high-capacity immobile Ca(2+) buffer (presumably plasma membrane associated) is suggested to be present in the soma and apical part of digits. Mitochondria also are likely to modulate synaptically evoked Ca(2+) responses in GrCs. The alternative hypotheses are thoroughly discussed and research avenues for their testing in future experiments are proposed. PMID- 19121635 TI - Role of down-regulated in adenoma anion exchanger in HCO3- secretion across murine duodenum. AB - BACKGROUND & AIMS: The current model of duodenal HCO(3)(-) secretion proposes that basal secretion results from Cl(-)/HCO(3)(-) exchange, whereas cyclic adenosine monophosphate (cAMP)-stimulated secretion depends on a cystic fibrosis transmembrane conductance regulator channel (Cftr)-mediated HCO(3)(-) conductance. However, discrepancies in applying the model suggest that Cl( )/HCO(3)(-) exchange also contributes to cAMP-stimulated secretion. Of 2 candidate Cl(-)/HCO(3)(-) exchangers, studies of putative anion transporter-1 knockout (KO) mice find little contribution of putative anion transporter-1 to basal or cAMP-stimulated secretion. Therefore, the role of down-regulated in adenoma (Dra) in duodenal HCO(3)(-) secretion was investigated using DraKO mice. METHODS: Duodenal HCO(3)(-) secretion was measured by pH stat in Ussing chambers. Apical membrane Cl(-)/HCO(3)(-) exchange was measured by microfluorometry of intracellular pH in intact villous epithelium. Dra expression was assessed by immunofluorescence. RESULTS: Basal HCO(3)(-) secretion was reduced approximately 55%-60% in the DraKO duodenum. cAMP-stimulated HCO(3)(-) secretion was reduced approximately 50%, but short-circuit current was unchanged, indicating normal Cftr activity. Microfluorimetry of villi demonstrated that Dra is the dominant Cl(-)/HCO(3)(-) exchanger in the lower villous epithelium. Dra expression increased from villous tip to crypt. DraKO and wild-type villi also demonstrated regulation of apical Na(+)/H(+) exchange by Cftr-dependent cell shrinkage during luminal Cl(-) substitution. CONCLUSIONS: In murine duodenum, Dra Cl(-)/HCO(3)(-) exchange is concentrated in the lower crypt-villus axis where it is subject to Cftr regulation. Dra activity contributes most basal HCO(3)(-) secretion and approximately 50% of cAMP-stimulated HCO(3)(-) secretion. Dra Cl(-)/HCO(3)(-) exchange should be considered in efforts to normalize HCO(3)(-) secretion in duodenal disorders such as ulcer disease and cystic fibrosis. PMID- 19121637 TI - Models of transcription factor binding: sensitivity of activation functions to model assumptions. AB - We present three models of how transcription factors (TFs) bind to their specific binding sites on the DNA: a model based on statistical physics, a Markov-chain model and a computational simulation. Comparison of these models suggests that the effect of non-specific binding can be significant. We also investigate possible mechanisms for cooperativity. The simulation model suggests that direct interactions between TFs are unlikely to be the main source of cooperativity between specific binding sites, because such interactions tend to lead to the formation of clusters on the DNA with undesirable side-effects. PMID- 19121638 TI - Mathematical and numerical analysis for a model of growing metastatic tumors. AB - In cancer diseases, the appearance of metastases is a very pejorative forecast. Chemotherapies are systemic treatments which aim at the elimination of the micrometastases produced by a primitive tumour. The efficiency of chemotherapies closely depends on the protocols of administration. Mathematical modeling is an invaluable tool to help in evaluating the best treatment strategy. Iwata et al. [K. Iwata, K. Kawasaki, N. Shigesad, A dynamical model for the growth and size distribution of multiple metastatic tumors, J. Theor. Biol. 203 (2000) 177.] proposed a partial differential equation (PDE) that describes the metastatic evolution of an untreated tumour. In this article, we conducted a thorough mathematical analysis of this model. Particularly, we provide an explicit formula for the growth rate parameter, as well as a numerical resolution of this PDE. By increasing our understanding of the existing model, this work is crucial for further extension and refinement of the model. It settles down the framework necessary for the consideration of drugs administration effects on tumour development. PMID- 19121640 TI - Antigenotoxic effect of apigenin against mitomycin C induced genotoxic damage in mice bone marrow cells. AB - The antigenotoxic effect of apigenin was studied against the genotoxic damage induced by mitomycin C on mouse bone marrow cells using sister chromatid exchanges (SCEs) and chromosomal aberrations (CAs) as parameters. Apigenin was studied at three different doses i.e. 10, 20 and 40 mg/kg b.w. and was found to be non-genotoxic at all the above three doses. Mitomycin C at 2 mg/kg b.w. was given along with the 10, 20 and 40 mg/kg bw of apigenin. A significant decrease in SCEs and CAs was observed, suggesting a protective role of apigenin against the genotoxicity of mitomycin C on mice bone marrow cells. PMID- 19121641 TI - The effect of sex hormones on bone metabolism of the otic capsule--an overview. AB - Bone resorption, which can occur after the menopause, has long been considered to due to the decrease of estrogen and so estrogen and estrogen/progestin treatment in women has been employed with the aim of slowing down the process. Other important factors have recently been considered, including follicle-stimulating hormone. The hormonal control of bone metabolism has taken on a new dimension since the description, within the last decade, of a major osteoclast inhibiting control system. The receptor activator of nuclear factor-kappaB (NF-kappaB) ligand (RANKL) produced by osteoblastic lineage cells, must bind with its receptor RANK, located on osteoclasts, in order to allow the maturation and activation of osteoclasts. The potential continuous bone loss is controlled by the decoy receptor osteoprotegerin (OPG) which competitively binds to RANKL and hence blocks the interaction of RANKL-RANK. Estrogen contributes to bone protection since it decreases the response of osteoclasts to RANKL and induces osteoclast apoptosis. But estrogen, alone and especially in synergy with progesterone, is a potent stimulator of prolactin release. Prolactin affects calcium metabolism and hyperprolactinemia associated with pregnancy, lactation, antipsychotic drug treatment, or aging is reflected in decreased bone mineral density. Long-term estrogen treatment in guinea pig results in hyperprolactinemia and has been shown to lead to hearing loss as well as bone dysmorphology of the otic capsule. Recent data show that prolactin decreases OPG and increases RANKL. OPG has been shown to be expressed at high levels in the cochlea and OPG knock out mice have indeed abnormal remodeling of the otic capsule and resorption of the auditory ossicles. So estrogen-induced hyperprolactinemia could oppose estrogen protection by the knock-down of the OPG bone protection system. This might explain why oral contraception treatment and hormone replacement therapies, involving estrogen together with progestin, increases the risk of otosclerosis and vestibular disorders. Hyperprolactinemia associated with pregnancy and lactation might also underlie the association of increased risk of otosclerosis with multiple pregnancies. PMID- 19121639 TI - Structural analysis of linear and conformational epitopes of allergens. AB - In many countries regulatory agencies have adopted safety guidelines, based on bioinformatics rules from the WHO/FAO and EFSA recommendations, to prevent potentially allergenic novel foods or agricultural products from reaching consumers. We created the Structural Database of Allergenic Proteins (SDAP, http://fermi.utmb.edu/SDAP/) to combine data that had previously been available only as flat files on Web pages or in the literature. SDAP was designed to be user friendly, to be of maximum use to regulatory agencies, clinicians, as well as to scientists interested in assessing the potential allergenic risk of a protein. We developed methods, unique to SDAP, to compare the physicochemical properties of discrete areas of allergenic proteins to known IgE epitopes. We developed a new similarity measure, the property distance (PD) value that can be used to detect related segments in allergens with clinical observed cross reactivity. We have now expanded this work to obtain experimental validation of the PD index as a quantitative predictor of IgE cross-reactivity, by designing peptide variants with predetermined PD scores relative to known IgE epitopes. In complementary work we show how sequence motifs characteristic of allergenic proteins in protein families can be used as fingerprints for allergenicity. PMID- 19121643 TI - Isolation, expression and characterization of two single-chain variable fragment antibodies against an endo-polygalacturonase secreted by Sclerotinia sclerotiorum. AB - Canola is a very important economic crop in the world and canola stem rot caused by Sclerotinia sclerotiorum (Lib.) de Bary, a necrotrophic, highly destructive and non-host-specific fungus, can reduce yield significantly. This fungus secretes numerous cell wall degrading enzymes including an endo polygalacturonase, SSPG1d, which has been detected at early stages of infection. In this report we describe the isolation of two recombinant antibodies of the single-chain variable fragment (ScFv) format from RNA of mice immunized with recombinant SSPG1d (rSSPG1d) or a peptide derived from SSPG1d (peptide 3796) that was predicted to be antigenic. The ScFvs were isolated using the established phage display technology. These recombinant antibodies were expressed, purified and refolded to functional antibodies with a yield of 120-500mug per liter of cell culture. Recombinant antibodies were characterized using various techniques including enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). Of the two ScFvs, it appears that only ScFv-rSSPG1d is able to detect whole SSPG1d produced by the fungus. Thus our results indicate that this ScFv may have utility in the detection of the SSPG1d enzyme in an antibody-based diagnostic test for S. sclerotiorum infection. PMID- 19121644 TI - Embryonic stem (ES) cell-derived cardiomyocytes: a good candidate for cell therapy applications. AB - During the last decade, embryonic stem cells (ESC) have unleashed new avenues in the field of developmental biology and emerged as a potential tool to understand the molecular mechanisms taking place during the process of differentiation from the embryonic stage to adult phenotype. Their uniqueness lies in retaining the capacity of unlimited proliferation and to differentiate into all somatic cells. Together with promising results from rodent models, ESC has raised great hope among for human ESC-based cell replacement therapy. ESC could potentially revolutionize medicine by providing a powerful and renewable cell source capable of replacing or repairing tissues that have been damaged in almost all degenerative diseases such as Parkinson's disease, myocardial infarction (MI) and diabetes. Somatic stem cells are an attractive option to explore for transplantation because they are autologous, but their differentiation potential is very limited. Currently, the major sources of somatic cells used for basic research and clinical trials come from bone marrow. But their widespread acceptability has not been gained because many of the results are confusing and inconsistent. The focus here is on human embryonic stem cells (hESCs), using methods to induce their differentiation to cardiomyocytes in vitro. Their properties in relation to primary human cardiomyocytes and their ability to integrate into host myocardium have been investigated into how they can enhance cardiac function. However, important aspects of stem cell biology and the transplantation process remain unresolved. In summary, this review updates the recent progress of ES cell research in cell therapy, discusses the problems in the practical utility of ESC, and evaluates how far this adjunctive experimental approach can be successful. PMID- 19121642 TI - NeuroD regulates proliferation of photoreceptor progenitors in the retina of the zebrafish. AB - neuroD is a member of the family of proneural genes, which function to regulate the cell cycle, cell fate determination and cellular differentiation. In the retinas of larval and adult teleosts, neuroD is expressed in two populations of post-mitotic cells, a subset of amacrine cells and nascent cone photoreceptors, and proliferating cells in the lineages that give rise exclusively to rod and cone photoreceptors. Based on previous studies of NeuroD function in vitro and the cellular pattern of neuroD expression in the zebrafish retina, we hypothesized that within the mitotic photoreceptor lineages NeuroD selectively regulates aspects of the cell cycle. To test this hypothesis, gain and loss-of function approaches were employed, relying on the inducible expression of a NeuroD(EGFP) fusion protein and morpholino oligonucleotides to inhibit protein translation, respectively. Conditional expression of neuroD causes cells to withdraw from the cell cycle, upregulate the expression of the cell cycle inhibitors, p27 and p57, and downregulate the cell cycle progression factors, Cyclin B1, Cyclin D1, and Cyclin E2. In the absence of NeuroD, cells specific for the rod and cone photoreceptor lineage fail to exit the cell cycle, and the number of cells expressing Cyclin D1 is increased. When expression is ectopically induced in multipotent progenitors, neuroD promotes the genesis of rod photoreceptors and inhibits the genesis of Muller glia. These data show that in the teleost retina NeuroD plays a fundamental role in photoreceptor genesis by regulating mechanisms that promote rod and cone progenitors to withdraw from the cell cycle. This is the first in vivo demonstration in the retina of cell cycle regulation by NeuroD. PMID- 19121645 TI - Human genetic determinants of dengue virus susceptibility. AB - Dengue virus (DENV) is an emerging mosquito-borne pathogen that produces significant morbidity worldwide resulting in an estimated 50-100 million infections annually. DENV causes a spectrum of illness ranging from inapparent infection to life-threatening hemorrhagic fever and shock. The varied DENV disease outcome is determined by complex interactions between immunopathologic, viral, and human genetic factors. This review summarizes these interactions with a focus on human genetic determinants of DENV susceptibility, including human leukocyte antigens, blood type, and single nucleotide polymorphisms in immune response genes that have been associated with DENV disease. We also discuss other factors related to DENV outcome including viral genetic determinants, age, ethnicity, and nutritional status as they relate to DENV susceptibility. We emphasize the need for functional genetics studies to complement association based data and we call for controlled study designs and standard clinical DENV disease definitions that will strengthen conclusions based on human genetic DENV studies. PMID- 19121647 TI - Clinical factors associated with liver stiffness in hepatitis B e antigen positive chronic hepatitis B patients. AB - BACKGROUND & AIMS: We analyzed the clinical factors associated with advanced liver fibrosis in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients. METHODS: We prospectively recruited treatment-naive HBeAg-positive patients for liver stiffness measurement (LSM) by transient elastography. Insignificant and advanced fibrosis was defined as an LSM of 6.0 kPa or less, and greater than 9.0 kPa for patients with alanine aminotransferase (ALT) levels less than or equal to the f upper limit of normal (ULN), and 7.5 kPa or less and greater than 9.0 kPa for those with ALT levels between 1 and 5 x ULN, respectively, based on a previous study with histologic validation. RESULTS: A total of 453 patients were studied. Among 74 patients who also had a liver biopsy, the cut-off levels for advanced fibrosis had 95% specificity. Age and ALT level, but not hepatitis B virus DNA level, were associated independently with LSM. Based on receiver operating characteristics curve analysis, patients older than 35 years had the highest specificity for advanced fibrosis. The risk of advanced fibrosis increased in patients with an ALT level greater than 0.5 x ULN. Among the 47 patients who were older than 35 years with an ALT level of 0.5 x ULN or less, 39 (83%) had an LSM suggestive of insignificant fibrosis, and 1 (2%) had advanced fibrosis. Among the 217 patients who were older than 35 years with an ALT level greater than 0.5 x ULN, 61 (28%) had LSM indicating insignificant fibrosis, and 80 (37%) had advanced fibrosis. CONCLUSIONS: Risk of advanced liver fibrosis increased in HBeAg-positive patients older than 35 years of age with an ALT level greater than 0.5 x ULN. PMID- 19121648 TI - Evidence of endothelial dysfunction in patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: Chronic inflammation has a major role in the development and propagation of endothelial dysfunction, which can lead to coronary artery disease. Endothelial dysfunction has been described in patients with various and diverse chronic inflammatory conditions. Altered vascular flow has been proposed to mediate inflammation in inflammatory bowel disease (IBD), although the role of endothelial dysfunction remains obscure. The purpose of our study was to assess endothelial function in patients with IBD. METHODS: Ninety-eight subjects were included in this study; 48 with IBD (17 with ulcerative colitis and 31 with Crohn's disease) and 50 healthy controls. Endothelial function was assessed by pulse arterial tonometry (PAT) and brachial ultrasound to determine flow-mediated dilation and shear stress reactive hyperemia. The impact of disease activity, disease duration, and IBD therapy also was assessed. RESULTS: Baseline demographic characteristics, including cardiovascular risk factors, were similar in all groups. IBD patients showed microvascular endothelial dysfunction, with lower PAT indices (P < .01) and shear stress reactive hyperemia (P < .05) compared with controls. There was no relationship between microvascular endothelial dysfunction, disease duration, underlying therapy, or clinical disease activity. There was a positive association between lower PAT scores and recent abdominal pain (P < .05). CONCLUSIONS: This was a large study assessing endothelial dysfunction in IBD. Both ulcerative colitis and Crohn's disease patients showed evidence of microvascular endothelial dysfunction. Future research could determine whether endothelial dysfunction is involved in the pathogenesis of IBD or increases the risk of cardiovascular events in this patient population. PMID- 19121649 TI - A retrospective single-center review of primary sclerosing cholangitis in children. AB - BACKGROUND & AIMS: Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by inflammation and progressive bile duct fibrosis. There are limited data on pediatric PSC. METHODS: We performed a retrospective chart review of 47 pediatric patients with PSC. RESULTS: The mean age at diagnosis was 11 +/- 4.9 years. Symptoms occurred before presentation in 81% of patients; inflammatory bowel disease was found in 59% and autoimmune hepatitis (overlap syndrome) in 25% of patients. Magnetic resonance cholangiography revealed both extrahepatic and intrahepatic, isolated intrahepatic, isolated extrahepatic, and no biliary involvement (small-duct PSC) in 40%, 14%, 10%, and 36%, respectively. Advanced fibrosis (stage >II) was present in 65%. Colonoscopy revealed pancolitis, rectal sparing, and normal findings in 24%, 24%, and 18%, respectively. All patients were treated with ursodeoxycholic acid (UDCA); 9 with overlap syndrome also received immunosuppressants. Fifteen patients without overlap syndrome had positive autoimmune markers and responded to UDCA monotherapy. Liver transplantation was performed in 9 patients (3 with overlap syndrome and 2 with small-duct PSC) at a median time of 7 years after diagnosis. The 10-year posttransplant survival rate was 89%. CONCLUSIONS: In one of the largest single-center studies of children with PSC, we found that most children with PSC had inflammatory bowel disease or autoimmune overlap and advanced fibrosis at diagnosis. Levels of alanine aminotransferase and gamma-glutamyl transferase were highest in patients with overlap syndrome and lowest in those with small-duct PSC. Levels of serum liver enzymes normalized after therapy with UDCA, including patients with positive autoimmune markers without histologic features of autoimmune hepatitis. PMID- 19121650 TI - Implementation of a data repository-driven approach for targeted proteomics experiments by multiple reaction monitoring. AB - Multiple reaction monitoring (MRM), commonly employed for the mass spectrometric detection of small molecules, is rapidly gaining ground in proteomics. Its high sensitivity and specificity makes this targeted approach particularly useful when sample throughput or proteome coverage limits global studies. Existing tools to design MRM assays rely exclusively on theoretical predictions, or combine them with previous observations on the same type of sample. The additional mass spectrometric experimentation this requires can pose significant demands on time and material. To overcome these challenges, a new MRM worksheet was introduced into The Global Proteome Machine database (GPMDB) that provided all of the information needed to design MRM transitions based solely on archived observations made by other researchers in previous experiments. This required replacing the precursor ion intensity by the number of peptide observations, which proved to be an adequate substitute if peptides did not occur in multiple forms. While the absence of collision energy information proved largely inconsequential, successful prediction of unique transitions depended on the type of fragment ion involved. The design of MRM assays for iTRAQ-labeled tryptic peptides obtained from human platelet proteins demonstrated the usefulness of the MRM worksheet also for quantitative applications. This workflow, which relies exclusively on experimental observations stored in data repositories, therefore represents an attractive alternative for the prediction of MRM transitions prior to experimental validation and optimization. PMID- 19121651 TI - Discussion: 'douching and the risk for sexually transmitted disease' by Tsai et al. AB - In the roundtable that follows, clinicians discuss a study published in this issue of the Journal in light of its methodology, relevance to practice, and implications for future research. Article discussed: Tsai CS, Shepherd BE, Vermund SH. Does douching increase risk for sexually transmitted infections? A prospective study in high-risk adolescents. Am J Obstet Gynecol 2009;200:38.e1 38.e8. PMID- 19121652 TI - Spontaneous rectovaginal fistula during bevacizumab therapy for ovarian cancer: a case report. AB - Recent studies have demonstrated efficacy of bevacizumab for recurrent ovarian cancer, but few data on its use and gastrointestinal potential complications when administered as adjuvant chemotherapy after cytoreductive surgery are available. In this study, we report the first case of a rectovaginal fistula in this indication. PMID- 19121653 TI - Vulvar hematoma secondary to spontaneous rupture of the internal iliac artery: clinical review. AB - Vulvar hematomas occur rarely outside the obstetric population but may present after other trauma to the pelvis or perineum. Spontaneous rupture of the internal iliac artery is described mostly in the presence of an aneurysm, with atherosclerosis, connective tissue disease, infection, and trauma as causative factors. It most often presents with abdominal pain and neurologic or urologic symptoms. We present an unusual case of a spontaneous rupture of the internal iliac artery that presented as a vulvar hematoma in a nulliparous woman that was successfully treated with selective arterial embolization and surgical evacuation. The literature is reviewed and management options discussed. PMID- 19121654 TI - Spatiotemporal electromyography during human labor to monitor propagation of the uterine contraction wave and diagnose dystocia. PMID- 19121655 TI - Saving lives and changing family histories: appropriate counseling of pregnant women and men and women of reproductive age, concerning the risk of diagnostic radiation exposures during and before pregnancy. AB - Over the past 50 years, our laboratory has provided consultations dealing with the risks of various environmental toxicant exposures during pregnancy. These contacts were primarily by telephone or written communications. Since the year 2000, the primary source of consultations has been via the internet. In 2007, the pregnancy website of the Health Physics Society received 1,299,672 visits. The contacts who downloaded information totaled 620,035. After reading the website information, 1442 individuals who were still concerned contacted me directly. Unfortunately, we have learned that many physicians and other counselors are not prepared to counsel patients concerning radiation risks. Approximately, 8% of the website contacts, who had consulted a professional, were provided inaccurate information that could have resulted in an unnecessary interruption of a wanted pregnancy. Research from our and other investigators' laboratories has provided radiation risk data that are the basis for properly counseling contacts with radiation exposures. Mammalian animal research has been an important source of information that improves the quality and accuracy of estimating the reproductive and developmental risks of ionizing radiation in humans. What are the reproductive and developmental risks of in utero ionizing radiation exposure? 1. Birth defects, mental retardation, and other neurobehavioral effects, growth retardation, and embryonic death are deterministic effects (threshold effects). This indicates that these effects have a no adverse effect level (NOAEL). Almost all diagnostic radiological procedures provide exposures that are below the NOAEL for these developmental effects. 2. For the embryo to be deleteriously affected by ionizing radiation when the mother is exposed to a diagnostic study, the embryo has to be exposed above the NOAEL to increase the risk of deterministic effects. This rarely happens when the pregnant women have x-ray studies of the head, neck, chest or extremities. 3. During the preimplantation and preorganogenesis stages of embryonic development, the embryo is least likely to be malformed by the effects of ionizing radiation because the cells of the very young embryo are omnipotential and can replace adjacent cells that have been deleteriously affected. This early period of development has been designated as "the all-or-none period." 4. Protraction and fractionation of exposures of ionizing radiation to the embryo decrease the magnitude of the deleterious effects of deterministic effects. 5. The increased risk of cancer following high exposures to ionizing radiation exposure to adult populations has been demonstrated in the atomic bomb survivor population. Radiation-induced carcinogenesis is assumed to be a stochastic effect (nonthreshold effect) so that there is theoretically a risk at low exposures. Whereas there is no question that high exposures of ionizing radiation can increase the risk of cancer, the magnitude of the risk of cancer from embryonic exposures following diagnostic radiological procedures is very controversial. Recent publications and analyses indicate that the risk is lower for the irradiated embryo than the irradiated child, which surprised many scientists interested in this subject, and that there may be no increased carcinogenic risk from diagnostic radiological studies. Examples of appropriate and inappropriate counseling will be presented to demonstrate how counseling can save lives and change family histories. The reader is referred to the Health Physics Society website to obtain many examples of the answers to questions posed by women and men who have been exposed to radiation (www.hps.org). Then click on ATE (ask the expert). PMID- 19121656 TI - To the point: reviews in medical education--the Objective Structured Clinical Examination. AB - This article, the eighth in the To the Point Series prepared by the Association of Professors of Gynecology and Obstetrics Undergraduate Medical Education Committee, discusses the effectiveness of the Objective Structured Clinical Examination (OSCE) for assessment of learners' knowledge, skills, and behaviors. The OSCE has also been used for the appraisal of residents and physicians undergoing licensure examinations; herein we focus on its application to undergraduate medical education. We review evidence for best practices and recommendations on effective use of the OSCE and requirements for and challenges to its implementation, including creative ways to design an OSCE program with a limited budget. We discuss its role in providing formative and summative feedback and describe learner performance on the OSCE as the OSCE relates to subsequent testing, including US Medical Licensing Examination step 1. A representative case with assessment used at the authors' medical schools is included. PMID- 19121657 TI - Spatiotemporal electrohysterography patterns in normal and arrested labor. AB - OBJECTIVE: The purpose of this study was to investigate the spatiotemporal patterns of uterine electrical activity in normal and arrested labors. STUDY DESIGN: From a database of electrohysterograms, 12 subjects who underwent cesarean delivery for active-phase arrest were each matched with 2 vaginally delivered controls. Using 30-minute segments of the electrohysterogram during the arrest, or the same dilation in controls, the center of uterine electrical activity was derived. The vertical motion of this center of uterine activity was determined for each contraction and the frequencies of movement patterns analyzed. RESULTS: Predominantly upward movement of the center of uterine activity (longer and/or stronger contraction at the fundus) was more common with normal dilation (P = .003). Receiver operating characteristic curve analysis gave an area under the curve of 0.91 for predicting outcome (vaginal vs cesarean delivery). CONCLUSION: There is a significant correlation between upward movement of the center of uterine activity (fundal dominance) and current labor progress. PMID- 19121658 TI - Assessment of the peak systolic velocity of the middle cerebral artery in twin twin transfusion syndrome. Part I: preoperative assessment. AB - OBJECTIVE: We sought to assess the incidence of an elevated peak systolic velocity of the middle cerebral artery (MCA-PSV) in twin-twin transfusion syndrome prior to laser surgery and its prognostic value for intrauterine fetal demise 24 hours after surgery (IUFD-24). STUDY DESIGN: An elevated MCA-PSV was defined as a velocity > .5 multiples of the median by transabdominal pulsed Doppler. Gestational age, Quintero stage, and number and/or type of placental vascular anastomoses were assessed as risk factors for an elevated MCA-PSV. Risk of IUFD-24 was assessed relative to an elevated MCA-PSV. RESULTS: An elevated MCA PSV was present in 4.2% of donors, 3.2% of recipients (P = .5), and 1.5% of both twins in 189 patients with twin-twin transfusion syndrome. An elevated MCA-PSV was unrelated to gestational age, stage, and number or type of anastomoses. An elevated MCA-PSV in the recipient twin was associated with an increased risk of IUFD-24 of this fetus (P = .01). CONCLUSIONS: An elevated MCA-PSV is present in < 5% of donor and recipient twins. An elevated MCA-PSV is a risk factor for IUFD-24 of the recipient twin. PMID- 19121659 TI - Breast cancer incidence after hormonal infertility treatment in Sweden: a cohort study. AB - OBJECTIVE: To assess the impact of infertility treatment with causes of infertility on incidence of breast cancer. STUDY DESIGN: Historical prospective cohort study of 1135 women attending major university clinics for treatment of infertility in Sweden, 1961-1976. Women were classified as users of clomiphene citrate or gonadotropins, or a combination of both therapies. Standardized incidence ratios were calculated to estimate relative risk of breast cancer. RESULTS: We observed 54 cases of breast cancer during 1961-2004, which did not significantly exceed those expected. Users of high-dose clomiphene citrate had an almost 2-fold increased risk (standardized incidence ratio, 1.90; 95% confidence interval, 1.08-3.35). This association was more pronounced among women referred for nonovulatory factors, with 3-fold increased risk (standardized incidence ratio, 3.00; 95% confidence interval, 1.35-6.67). CONCLUSION: No overall increased risk for breast cancer was shown with infertility treatment. Women with nonovulatory causes treated with high-dose clomiphene citrate therapy may have an elevated risk for breast cancer. PMID- 19121660 TI - The adaptor protein MyD88 is essential for E coli-induced preterm delivery in mice. AB - OBJECTIVE: We used a mouse model of infection-induced preterm delivery to examine the roles of 2 adaptor proteins with central functions in Toll-like receptor signaling: MyD88 (myeloid differentiation primary-response gene 88) and TRIF (Toll/IL-1 receptor (TIR)-domain-containing adaptor protein-inducing IFN-beta). STUDY DESIGN: Mice deficient (KO) for MyD88, TRIF, both (DKO) or neither (WT) were inoculated into the uterus with killed Escherichia coli. Delivery outcomes, fetal status, serum progesterone, and nuclear translocation of the transcription factor nuclear factor kappa B (NFkappaB) were determined. RESULTS: Preterm birth (delivery in less than 48 hours) occurred in WT and TRIF-KO animals in a dose dependent fashion, reaching 100% with 5-10 x 10(9) bacteria, while MyD88-KO and DKO animals were completely protected from delivery. Intrauterine fetal survival, maintenance of circulating progesterone levels, and nuclear translocation of NFkappaB were also dependent upon MyD88 but not TRIF. In contrast, induction of uterine interleukin (IL)-1beta and tumor necrosis factor alpha (TNF-alpha) depends upon actions of both MyD88 and TRIF. CONCLUSION: E coli-induced preterm delivery in the mouse is completely dependent upon MyD88 but not TRIF. PMID- 19121661 TI - Ureaplasma colonization of amniotic fluid and efficacy of antenatal corticosteroids for preterm lung maturation in sheep. AB - OBJECTIVE: The objective of the study was to assess the efficacy of maternal betamethasone for improving preterm lung function in the presence of inflammation induced by amniotic fluid Ureaplasma colonization. STUDY DESIGN: Ewes bearing single fetuses were randomized to receive an intraamniotic injection of Ureaplasma parvum (serovar 6; 2 x 10(7) colony-forming units) or vehicle at 86 +/ 2 days of pregnancy (mean +/- SD: term is 150 days), followed by maternal intramuscular betamethasone (0.5 mg/kg) or saline, either 2 or 7 days before delivery of lambs at 123 +/- 1 d. RESULTS: Amniotic fluid interleukin-8 was elevated by ureaplasmas (P = .049) but unaffected by betamethasone. Lung inflammation induced by ureaplasmas was not affected by betamethasone. Lung compliance was increased by Ureaplasma colonization (P = .009) and betamethasone (P = .042), and effects were additive. Lung surfactant was increased by Ureaplasma colonization (P < .001) and betamethasone 7 days (P = .001), but not 2 days, before delivery. CONCLUSION: Inflammation improves preterm lung function because of increases in surfactant. Antenatal corticosteroids further augment lung function through an apparently independent mechanism. PMID- 19121662 TI - Quantification of cell free fetal DNA in maternal plasma in normal pregnancies and in pregnancies with placental dysfunction. AB - OBJECTIVE: To assess the normal levels of free fetal DNA in maternal plasma through pregnancy compared with those in pregnancies complicated with placental dysfunction manifested by preeclampsia and/or fetal growth restriction. STUDY DESIGN: Maternal blood samples from 138 singleton male pregnancies were divided into 3 groups; normal pregnancies (77), preeclampsia (49), and fetal growth restriction (12). Royston and Wright's methods were used to calculate gestational age-related reference limits of free fetal DNA in the 3 groups. The DYS14 gene of the Y chromosome was quantified and compared in maternal plasma by using real time quantitative polymerase chain reaction. RESULTS: Free fetal DNA in normal pregnancies increased with gestational age. Results were significantly higher in preeclampsia and fetal growth restriction groups than in normal pregnancy and were higher in severe preeclampsia than in milder disease. CONCLUSION: Free fetal DNA is a potential marker for placental dysfunction in pregnancy. Large prospective studies are now needed to investigate its role in the prediction of pregnancy complications and severity and or timing of delivery. PMID- 19121663 TI - Term labor is associated with a core inflammatory response in human fetal membranes, myometrium, and cervix. AB - OBJECTIVE: Identify overlap of the transcriptome of myometrium and cervix in association with human labor. STUDY DESIGN: The transcriptional profile of myometrial and cervical biopsies obtained from women in spontaneous labor at term (n = 9) and not in labor (n = 9) were characterized by Affymetrix v2 U133 plus 2 arrays. Common canonical pathways and functional groups were identified by Ingenuity Pathway Analysis. RESULTS: One hundred ten genes (false discovery rate < 1%) were commonly up-regulated by myometrium and cervix in association with labor and 29 genes (false discovery rate < 1%) down-regulated. Fold change in expression of up-regulated genes was strongly correlated; myometrium vs cervix (r = 0.51; P < .001), with no relationship in down-regulated genes (r = 0.26; P = .16). Canonical pathway analysis established up-regulation of inflammatory pathway signaling, with greatest increases in cellular movement and immune response gene ontology groups. CONCLUSION: Gestational tissues exhibit a core inflammatory response in association with human parturition, with pathways regulating cellular trafficking dominating. PMID- 19121664 TI - Douching and the risk for sexually transmitted disease: Tsai et al. AB - The article below summarizes a roundtable discussion of a study published in this issue of the Journal in light of its methodology, relevance to practice, and implications for future research. Article discussed: Tsai CS, Shepherd BE, Vermund SH. Does douching increase risk for sexually transmitted infections? A prospective study in high-risk adolescents. Am J Obstet Gynecol 2009;200:38.e1 38.e8. PMID- 19121665 TI - Complex abdominal wall reconstruction after radiation therapy: a full-thickness defect was repaired with a rectus femoris myofasciocutaneous flap. PMID- 19121666 TI - The global distribution of Burkholderia pseudomallei and melioidosis: an update. AB - While Southeast Asia and northern Australia are well recognized as the major endemic regions for melioidosis, recent reports have expanded the endemic zone. Severe weather events and environmental disasters such as the 2004 Asian tsunami have unmasked locations of sporadic cases and have reconfirmed endemicity in Indonesia. The endemic region now includes the majority of the Indian subcontinent, southern China, Hong Kong and Taiwan. Sporadic cases have occurred in Brazil and elsewhere in the Americas and in island communities such as New Caledonia, in the Pacific Ocean, and Mauritius in the Indian Ocean. Some of the factors that are critical to further elucidating the global distribution of Burkholderia pseudomallei and melioidosis include improved access to diagnostic laboratory facilities and formal confirmation of the identity of bacterial isolates from suspected cases. PMID- 19121667 TI - Re-emergence of melioidosis in Myanmar. AB - After the discovery of melioidosis and its causative organism, Burkholderia pseudomallei, in Burma (Myanmar) in 1912 and further reports in 1915 and 1917, no further reports of melioidosis emerged from the country in the international medical literature. The present study aimed to determine whether the occurrence of melioidosis among hospital patients with abscess in Yangon, Myanmar is sufficiently frequent to indicate further studies to determine its clinical and public health significance in the country. A clinical and bacteriological survey of patients with abscess at 21 hospitals in Yangon was carried out during 2004 2006. Pus specimens from three of 133 patients yielded bacteria with the morphological, cultural and biochemical characteristics of B. pseudomallei. The estimated melioidosis prevalence rate among patients with abscess in selected Yangon hospitals was approximately 2%. Further studies are indicated to assess the clinical and public health significance of melioidosis in Myanmar. PMID- 19121668 TI - Melioidosis: antibiogram of cases in Singapore 1987-2007. AB - The antimicrobial susceptibility of Burkholderia pseudomallei isolates identified by the bacteriology laboratory at Singapore General Hospital was reviewed. Laboratory data were found to be available since 1987 and showed that ceftazidime, amoxicillin-clavulanate, chloramphenicol and tetracycline had remained effective through the years. Imipenem was added to the list of antimicrobials tested after 1989, and the isolates showed high susceptibility rates. Co-trimoxazole was found to be useful based on Etest results, but the isolates had low susceptibility rates when tested using disk diffusion. PMID- 19121669 TI - High-level resistance to fluoroquinolones and cephalosporins in Burkholderia pseudomallei and closely related species. AB - The molecular mechanisms involved in the development of a high level of resistance to a wide range of antimicrobials in Burkholderia pseudomallei and closely related species have not been sufficiently investigated. In the present study, the properties of B. pseudomallei, B. mallei and B. thailandensis mutants with increased resistance to fluoroquinolones and cephalosporins were analysed. Resistance to pefloxacin, ofloxacin and ceftazidime in B. pseudomallei and B. thailandensis was accompanied by an increased resistance to aminoglycosides, beta lactams, macrolides and chloramphenicol, whereas mutants of B. mallei were characterized by a narrower spectrum of resistance. With the use of the differential display technique, we demonstrated that multiple resistant variants of B. pseudomallei, B. mallei and B. thailandensis had an increased expression of putative efflux transporters belonging to the resistance-nodulation division superfamily and the major facilitator superfamily. With the application of PCR single-stranded conformational polymorphism (PCR-SSCP) and sequencing, point mutations in gyrA quinolone-resistance determining region were detected in the part of multiple resistant B. pseudomallei and B. mallei mutants. These results indicate that various molecular mechanisms are involved in the development of multiple drug resistance in pathogenic Burkholderia and may be useful for further studying the adaptability of this microorganism and optimization of treatment. PMID- 19121670 TI - Burkholderia pseudomallei: animal models of infection. AB - A range of animal models of Burkholderia pseudomallei infection have been reported, and the host species differ widely both in their susceptibility to infection and in the pathogenesis of disease. In mice, and depending on the route of infection, dose, and mouse strain, the disease can range from a chronic, and in some cases, an apparently latent infection to an acute fulminant disease. Alternative small animal models of infection include diabetic rats or hamsters. Larger animal models of disease have not yet been fully developed. It is not clear which of the small animal models of melioidosis most accurately reflect disease in humans. However, the findings that diabetic rats are susceptible to infection, that some strains of mice can develop persistent subclinical infections that can spontaneously reactivate, and that inhalation exposure generally results in more acute disease suggest that these different models mimic different aspects of human melioidosis. PMID- 19121671 TI - Melioidosis masquerading as enteric fever. AB - Melioidosis is endemic in Southeast Asia and northern Australia, but it has been rarely reported from India. Recent reports have shown that melioidosis is an emerging infection in this part of the world, but enteric fever is more commonly seen in India. We present a 50-year-old male with diabetes who presented with acute onset of febrile illness. Preliminary investigations were suggestive of enteric fever, and he showed a partial response to parenteral ceftriaxone; however, it later turned out that he had melioidosis. The widal titres were persistently elevated even following treatment with meropenem. PMID- 19121672 TI - Inhalation of Burkholderia thailandensis results in lethal necrotizing pneumonia in mice: a surrogate model for pneumonic melioidosis. AB - Burkholderia thailandensis is closely related to B. pseudomallei, the causative agent of melioidosis, a lethal tropical disease. Burkholderia thailandensis is largely avirulent in humans and, unlike B. pseudomallei, does not require strict biocontainment conditions for study. Because it may be a useful research surrogate for B. pseudomallei, we developed a murine model of airborne B. thailandensis infection. In both C57BL/6 and BALB/c mice, deposition of 103 colony-forming units (CFU)/lung or less of B. thailandensis was non-lethal and infection was readily controlled. Compared with C57BL/6 mice, BALB/c mice exhibited modest resistance to infection after deposition of 10(4) CFU/lung. Deposition of 10(5) CFU/lung resulted in disseminated infection and was universally fatal by 3d. This dose induced robust pulmonary neutrophilia, production of inflammatory cytokines, and elevated serum markers of distant organ injury. Histology demonstrated multiple small foci of necrotizing pneumonia but lung architecture was otherwise preserved, suggesting that respiratory failure is not the cause of death. These findings demonstrate that airborne B. thailandensis infection in mice provides an accessible surrogate model of melioidosis. PMID- 19121673 TI - Emergence of Burkholderia pseudomallei and pandrug-resistant non-fermenters from southern Karnataka, India. AB - Melioidosis has recently gained the status of an emerging disease in India. Multidrug-resistant (MDR) Gram-negative bacteria, however, are already responsible for treatment failure and mortality. In addition, pandrug-resistant (PDR) Gram-negative bacteria have emerged as a new threat to modern medicine. The treatment, clinical follow up, and outcome of 25 patients with melioidosis and 46 patients with MDR non-fermenters (Pseudomonas aeruginosa and Acinetobacter spp.) infection were documented during the period 2005 2007. Pandrug resistance status of the MDR strains was evaluated with the minimum inhibitory concentration breakpoint of colistin. Skin and soft-tissue involvement (16%), liver abscess (16%) and bone and joint involvement (16%) were the most common presentations of melioidosis in diabetic patients. The presence of septicaemia (44%) and major organ failure (48%) resulted in death. Relapse was seen in patients with inappropriate treatment. Clinical cure was observed in five cases infected with PDR strains; colistin was used in only one case. Comorbid conditions may have contributed to the high fatality (82.7%). More awareness among clinicians and laboratory staff, and environmental investigations of soil are required for accurate diagnosis and prompt treatment of melioidosis. For MDR strains, colistin is the 'last resort' and should be used with caution; resistance should be monitored both globally and locally. PMID- 19121674 TI - Construction of a reporter system to study Burkholderia mallei type III secretion and identification of the BopA effector protein function in intracellular survival. AB - Burkholderia mallei, the aetiological agent of glanders disease, is a Gram negative facultative intracellular bacterium. Despite numerous studies, the detailed mechanism of its pathogenesis is almost unknown. The presence of a type III secretion system (TTSS) is one of the known mechanisms associated with virulence. An intact TTSS indicates that B. mallei is able to secrete proteins in response to different environmental conditions, which could play an important role in pathogenesis. Therefore, characterization of the TTSS and identification of the secreted proteins associated with bacterial pathogenesis could provide crucial information for the development of a candidate vaccine. In the current study, we used an enzymatic reporter system to establish some of the conditions enabling TTS. Construction of the TTSS bopA mutant revealed that BopA is important for B. mallei invasion and intracellular survival. Overall, our study elucidates how BopA can aid in the optimization of TTS and defines the function of TTS effectors in bacterial intracellular survival and invasion. PMID- 19121675 TI - Molecular identification and typing of Burkholderia pseudomallei and Burkholderia mallei: when is enough enough? AB - Burkholderia mallei and B. pseudomallei are highly pathogenic microorganisms for both humans and animals. Moreover, they are regarded as potential agents of bioterrorism. Thus, rapid and unequivocal detection and identification of these dangerous pathogens is critical. In the present study, we describe the use of an optimized protocol for the early diagnosis of experimental glanders and melioidosis and for the rapid differentiation and typing of Burkholderia strains. This experience with PCR-based identification methods indicates that single PCR targets (23S and 16S rRNA genes, 16S-23S intergenic region, fliC and type III secretion gene cluster) should be used with caution for identification of B. mallei and B. pseudomallei, and need to be used alongside molecular methods such as gene sequencing. Several molecular typing procedures have been used to identify genetically related B. pseudomallei and B. mallei isolates, including ribotyping, pulsed-field gel electrophoresis and multilocus sequence typing. However, these methods are time consuming and technically challenging for many laboratories. RAPD, variable amplicon typing scheme, Rep-PCR, BOX-PCR and multiple-locus variable-number tandem repeat analysis have been recommended by us for the rapid differentiation of B. mallei and B. pseudomallei strains. PMID- 19121676 TI - A role of Burkholderia pseudomallei flagella as a virulent factor. AB - Burkholderia pseudomallei is an agent of melioidosis and is closely related to avirulent B. thailandensis. Burkholderia thailandensis has a 15-bp deletion within the variable region of the flagellin gene fliC compared with B. pseudomallei. The difference in the fliC gene might be related to virulence. In the present study, the invasion, internalization and intracellular replication of both phagocytic (mouse macrophage cell line RAW264.7) and non-phagocytic cells (human lung epithelial cell line A549) of B. pseudomallei fliC knockout mutant (MM35) complemented with its own fliC (Cp) or with B. thailandensis fliC (Ct) was compared with those of the wild-type strains of B. pseudomallei (1026b) and B. thailandensis (E257). In phagocytic cells, there was no significant difference in bacterial uptake between Cp and Ct, but MM35 was internalized significantly less compared with 1026b, Cp, Ct and E257. The results suggest that flagella are involved in macrophage invasion. In non-phagocytic cells, Cp and Ct showed similar invasive capacities while 1026b, Cp and Ct showed significantly higher invasiveness than MM35, suggesting that flagella facilitate the non-phagocytic cell invasion. However, the invasive capacity of MM35 was significantly higher than that of E257, suggesting that in addition to the flagella, B. pseudomallei may need other factor(s) to facilitate invasion in non-phagocytic cells. PMID- 19121677 TI - Expression of resistance-nodulation-cell-division efflux pumps in commonly used Burkholderia pseudomallei strains and clinical isolates from northern Australia. AB - Burkholderia pseudomallei is the aetiological agent of melioidosis. Therapy for this disease is lengthy and limited to only a few antibiotics because of this bacterium's intrinsic antibiotic resistance to many clinically useful antibiotics. These properties of B. pseudomallei may partially be due to expression of efflux pumps of the resistance-nodulation-cell-division (RND) family. The patterns and magnitude of RND efflux pump expression in commonly used strains and clinical isolates of B. pseudomallei from the Royal Darwin Hospital, Darwin, Australia, were assessed in cells grown to late exponential phase using quantitative real-time PCR (qRT-PCR). Expression of the three previously identified RND efflux pumps AmrAB-OprA, BpeAB-OprB and BpeEF-OprC, as well as four other yet uncharacterized pumps, was found to be widespread in the clinical isolates. In 45 of 50 isolates (90%), mRNA was detected for at least one of the seven RND pumps. Of these 45 isolates, 41 (82%) expressed multiple pumps with nine strains expressing all seven pumps tested. While these studies revealed no striking correlation between RND efflux pump expression and clinically significant antibiotic resistance, the data support the notion that RND pumps probably play important roles in this bacterium's physiology, defence against toxic compounds, and perhaps virulence. PMID- 19121679 TI - Melioidosis--a case series from south India. AB - Melioidosis is sporadically reported from various parts of India. We present a case series from south India, highlighting the varied manifestations of the disease. Seven cases of culture-proven melioidosis are presented in whom Burkholderia pseudomallei were isolated from aspirate or blood. Melioidosis had a varied presentation involving multiple abscesses in the soft tissues, liver, spleen, mediastinum and the subdural space. It presented as either acute fulminant sepsis or followed a chronic indolent course, mimicking tuberculosis. Most cases had predisposing risk factors such as diabetes and chronic alcoholism. PMID- 19121678 TI - Cellular reporter screens for inhibitors of Burkholderia pseudomallei targets in Pseudomonas aeruginosa. AB - To facilitate the discovery of new therapeutics for Burkholderia pseudomallei infections, we have developed cellular reporter screens for inhibitors of B. pseudomallei targets in the surrogate host Pseudomonas aeruginosa. Pseudomonas aeruginosa strains carrying deletions of essential genes were engineered to be dependent on the isopropyl-beta-D-thiogalactopyranoside (IPTG)-regulated expression of their B. pseudomallei orthologues on a broad-host-range plasmid. Pseudomonas aeruginosa genes which are upregulated in response to depletion of each target gene product, were fused to the Photorhabdus luminescens luxCDABE operon via pGSV3-lux-Sp(R) to generate reporter strains with increased bioluminescence upon target inhibition. A total of 11 of 19 B. pseudomallei genes complemented deletions of their orthologues in P. aeruginosa. The dependence of growth on IPTG levels varied from complete dependence (ftsQ, gyrA, glmU, secA) to slower growth in the absence of IPTG (coaD, efp, mesJ), to apparently normal growth in the absence of IPTG (ligA, lpxA, folA, ipk). Reporter screening strains have been constructed for three gene targets (gyrA, glmU, secA), and one (gyrA) has been applied to 68,000 compounds resulting in a primary hit rate of 0.5% and a confirmed hit rate of 0.06%, including several fluoroquinolones. These results provide proof of principle for surrogate cellular reporter screens as a useful approach to identify inhibitors of essential gene products. PMID- 19121680 TI - A first report of pulmonary melioidosis in Cambodia. AB - Melioidosis has never been officially reported from Cambodia. Here we report two cases, a 58-year-old male (case 1) and a 49-year-old female (case 2) who presented with respiratory illnesses featuring multiple lung abscesses. The sputum culture of both patients, taken in the framework of a laboratory-based study on aetiologies of (sub-)acute respiratory infections among hospitalized patients in southern Cambodia, grew Burkholderia pseudomallei. The most striking aspect of these case stories was the extent of the delays in diagnosis. Presenting with a 1-month history of respiratory symptoms, case 1 was first suspected of tuberculosis (TB) infection, and then misdiagnosed as 'metastatic lung cancer' in Phnom Penh, Cambodia. Case 2 suffered from pulmonary infections for >10 years, during which time she was treated for TB four times. Neither patient ever produced acid-fast-bacilli (AFB)-positive sputum. Following our laboratory confirmation, the patients were traced for re-admission. Under the 'classical' trimethoprim sulphamethoxazole, chloramphenicol and doxycycline treatment, their clinical status improved considerably within 2 weeks. The two study cases illustrate issues relating to the misdiagnosis of melioidosis in Cambodia; an unfamiliarity of clinicians with the disease, which is associated with a high prevalence of TB. Therefore, a heightened awareness of melioidosis among clinicians would have a substantial impact on public health as the non septicaemic form of the disease is potentially treatable with antibiotics that are available in Cambodian public hospitals. PMID- 19121681 TI - Melioidosis and Aboriginal seasons in northern Australia. AB - Melioidosis, an infection due to the environmental bacterium Burkholderia pseudomallei, is endemic to Southeast Asia and northern Australia, with cases strongly correlated with the monsoonal wet season. We hypothesized that seasonal variation in the mode of acquisition, informed by traditional knowledge, would result in variations in disease characteristics as well as disease incidence. We explored the seasonal variation in acute, culture-confirmed melioidosis using local Aboriginal definitions of seasons in presentations to the Royal Darwin Hospital, the referral centre for the Top End of the Northern Territory, Australia. In 387 patients, we observed an increased proportion of patients with pneumonia (60%) and severe sepsis (25%) associated with presentations in the wet seasons Gunumeleng (October-December) and Gudjewg (January-March) compared with the drier seasons Wurrgeng (June August) and Gurrung (August-October) (pneumonia 26%, severe sepsis 13%). This observation supports the hypothesis that in the wet seasons there may be changes in the mode and/or magnitude of exposure to B. pseudomallei, with a shift from percutaneous inoculation to aerosol inhalation, for instance. PMID- 19121682 TI - Clinical and microbiological features of melioidosis in northern Vietnam. AB - Sporadic cases of melioidosis have been reported from Vietnam for decades, but clinical and epidemiological data for the indigenous population are still scarce. In this study, we reviewed clinical and demographic data of patients with culture proven melioidosis diagnosed at a single large referral hospital in Hanoi between November 1997 and December 2005. We found that the clinical manifestations of melioidosis (with fatal septicaemia as the most common presentation), a high rate of underlying diseases, and a peak of cases admitted during the wet season, were similar to studies from other endemic areas. The geographical origin of patients with melioidosis showed that melioidosis existed in at least 18 northern provinces. The characterization of clinical Burkholderia pseudomallei strains by multilocus sequence typing identified 17 different sequence types (STs), 11 of which have (as yet) not been found outside Vietnam. Several of these STs presumably were generated through recent evolutionary events in this rapidly diversifying bacterial species, and thus, restricted geographic distribution may be a consequence of limited time passed since emergence. To our knowledge, this is the first report on a series of cases describing clinical and epidemiological features of melioidosis and corresponding B. pseudomallei strains from northern Vietnam. PMID- 19121683 TI - Intensity of exposure and incidence of melioidosis in Thai children. AB - There is a high background seroprevalence of antibodies to Burkholderia pseudomallei in Thailand that limits its use as a diagnostic tool. It is believed that this results from childhood exposure to the bacterium in mud and surface water. The increasing prevalence of antibodies with age is a marker of the intensity of exposure. A susceptible-infected-susceptible (SIS) model was calibrated with data on seroprevalence in children (<15 years) in Udon Thani and Ubon Ratchathani (n=2214). In this mathematical model, children were assumed to gain antibodies at a constant rate related to exposure events, and waning antibody response occurred at a constant rate. The intensity of exposure appeared to be higher in Udon Thani than in Ubon Ratchathani, with 24% vs. 11% of patients becoming seropositive each year. In Udon Thani children, antibodies appeared to be long-lasting, compared with those in Ubon Ratchathani, where the mean duration was 5.2 years. Based on an estimated paediatric disease incidence in Ubon Ratchathani of 4.15 per 100,000 population, it is estimated that approximately 1 in 4600 antibody-producing exposures results in clinical infection. Childhood seroprevalence can be used as a marker of intensity of exposure. Further work to separate the effect of exposure to B. thailandensis and cross-reactivity to B. pseudomallei is proposed. PMID- 19121684 TI - 'Imported' melioidosis in Germany: relapse after 10 years. AB - A 62-year-old German patient with insulin-dependent diabetes and diverticulitis was hospitalized for abdominal pain of the left lower quadrant. Further examination revealed an abdominal abscess, which was punctured. Presumptively a Pseudomonas species was identified, but further examination revealed Burkholderia pseudomallei as the causative agent. Most probably this infection was acquired in 1996 during a trip to Thailand, where the patient had been hospitalized. After combined chemotherapy and surgical revision of the abscess, the patient's condition improved. Clinicians and microbiologists have to keep in mind that in some tropical infections such as melioidosis relapse may occur after such a long time. PMID- 19121685 TI - Burkholderia pseudomallei identification: a comparison between the API 20NE and VITEK2GN systems. AB - Sixty unique clinical isolates previously reported as Burkholderia pseudomallei were run in parallel through the API 20NE and VITEK2GN identification systems after 24h growth on Columbia agar with 5% sheep blood. The identifications and confidence levels determined by each test modality were compared. The VITEK2GN identified 47 (78.3%) and the API20NE identified 52 (86.7%) of the isolates as B. pseudomallei. The modal octal profile for the latter was 1156577. The alternative identification most commonly made by both systems was B. cepacia. Comparison of the two modalities gave a Cohen's kappa of 0.6008, suggesting good overall inter test agreement. Both are good test modalities capable of identifying B. pseudomallei reliably, with the API20NE having the advantage of correctly identifying a slightly greater number of isolates, and the VITEK2 having the advantage of a shorter test turnaround time. PMID- 19121686 TI - Validation and comparison of an extrapolysaccharide (EPS)-based in-house ELISA and the PanBio melioidosis rapid cassette test-kits for serodiagnosis of melioidosis in a non-endemic area. AB - Detection of anti-Burkholderia pseudomallei antibodies in sera from melioidosis patients still represents a keystone in the confirmation of the clinical diagnosis, especially in non-endemic areas. An in-house assay was compared to lateral flow assays for the rapid detection of melioidosis-specific IgG or IgM. Employing 50 positive sera from patients and 200 negative sera from blood donors, sensitivity of the ELISA, the IgG and IgM assay were 84.0%, 90.0% and 84.0%, respectively. Specificity ranged from 98.0% (ELISA) to 99.5% (IgM assay). The application of the described diagnostic assays is a suitable method for the serodiagnosis of melioidosis in a non-endemic area. PMID- 19121687 TI - Mimotope identification from monoclonal antibodies of Burkholderia pseudomallei using random peptide phage libraries. AB - This study used random peptide libraries, displayed by bacteriophage T7 and M13, to identify mimotopes from four monoclonal antibodies (mAbs) specific to Burkholderia pseudomallei. Bound phages, selected from fourth-round panning with each mAb, were tested for binding specificity with each mAb using ELISA, before being further amplified and checked for phage peptide sequence using PCR and DNA sequencing. Overall, 75 of 90 phages (83.3%) were ELISA-positive with each mAb. Mimotopes from all 75 phages (100%) were found to match protein sequences of Burkholderia spp. from GenBank. The predominant mimotopes were TP-GRTRVT found in 13.3%, LTPCGRTxD (8%), AREVTLL (6.7%), NxVxKVVSR (5.3%), PCAPRSS (4%), LGRVLAN (4%), RNPKKA (2.7%) and CPYPR (2.7%). The following GenBank-matched proteins (i.e. the hypothetical proteins) were located at the outer membrane of Burkholderia spp.: BPSL2046 of B. pseudomallei K96243 (matched with mimotope CGRTxD), BpseP_02000035 (matched with LGRVLAN), BPSS0784 of B. pseudomallei K96243 (matched with CPYPR), BURPS1710b_1104 of B. pseudomallei 1710b (matched with CARQY) and TonB-dependent siderophore receptor of B. cenocepacia H12424 (matched with CVRxxLTPC and TPCRxRT). These phage mimotopes and matched proteins may have the potential for further use as diagnostic reagent and immunogen against melioidosis. These results demonstrate that phage-display technique has the potential for rapidly identifying phage mimotopes that interact with B. pseudomallei mAbs. PMID- 19121688 TI - Soil physicochemical properties related to the presence of Burkholderia pseudomallei. AB - The incidence of Burkholderia pseudomallei in the soil from north-east Thailand is estimated to be 20-fold higher than that from central Thailand, and is associated with a 10-fold higher incidence of melioidosis in the region than in central Thailand. This study investigated the presence of B. pseudomallei in relation to the physicochemical properties of soil from Khon Kaen province, north east Thailand. Thirteen districts (54.2%) were positive for B. pseudomallei. From a selected district, B. pseudomallei was cultured from 19 of 50 sites (38%). The soil in this area was predominantly sandy. From the positive sites, the organism was found mainly at a depth of 30 cm (43/68, 63% of isolates) and was significantly associated with certain soil physicochemical parameters, including a pH of 5.0-6.0, a moisture content >10%, and higher chemical oxygen demand and total nitrogen than negative sites (P<0.05). Burkholderia pseudomallei is unevenly distributed in this area, with the pH of the soil being the major determinant of the presence of the organism. The sandy soil type of north-east Thailand can support the survival of B. pseudomallei and allow it to move freely with water flow, and thus readily come in contact with people during the rainy season. PMID- 19121689 TI - Development of aptamers specific for potential diagnostic targets in Burkholderia pseudomallei. AB - Improved diagnostic reagents would be of considerable benefit in enhancing the specificity and sensitivity of rapid assays for Burkholderia pseudomallei, the causative agent of melioidosis. The purpose of this work is to develop aptamers, high affinity RNA-based molecular recognition molecules, which could be used as reagents for identification of the whole organism in assays of biological samples. Data are presented demonstrating the purification of recombinant B. pseudomallei secreted or surface-exposed macromolecules, which have been expressed in Escherichia coli, and the initial stages of aptamer generation using these recombinant proteins. Future studies will focus upon the expansion of this methodology to include other target macromolecules located on or near the outer membrane of this organism. PMID- 19121690 TI - Sero-characterization of lipopolysaccharide from Burkholderia thailandensis. AB - We report the successful purification of lipopolysaccharide (LPS) from Burkholderia thailandensis, a Gram-negative bacterium, closely related to the highly pathogenic organisms B. pseudomallei and B. mallei. Burkholderia thailandensis LPS is shown to cross-react with rabbit and mouse sera obtained from inoculation with B. pseudomallei or B. mallei, respectively. These data suggest that B. thailandensis LPS shares similar structural features with LPS molecules from highly pathogenic Burkholderia species. This information may prove useful in ongoing efforts to develop novel vaccines and/or diagnostic reagents. PMID- 19121691 TI - In silico analysis of potential diagnostic targets from Burkholderia pseudomallei. AB - Administration of appropriate therapeutic regimes for infections arising from pathogenic species of Burkholderia is critically dependent upon rapid and accurate diagnoses. The purpose of this work is to establish a bioinformatic pipeline to assess protein sequences for their potential as diagnostic targets for the detection of Burkholderia species. Data are presented showing both a bioinformatic methodology for prediction of surface-associated and secreted proteins and its application to a test dataset of proteins from the pathogen B. pseudomallei. A subset of proteins, known to be produced by the organism, is identified which represents potential targets for development of new diagnostic reagents. In addition, a 'reverse diagnostics' bioinformatics approach has been established which can now be extended to whole genome analyses. PMID- 19121692 TI - Seropositivity to Burkholderia pseudomallei does not reflect the development of cell-mediated immunity. AB - Cell-mediated immunity to Burkholderia pseudomallei, the causative agent of melioidosis, provides protection from disease progression. An indirect haemagglutination assay was used to detect antibodies to B. pseudomallei in 1500 healthy donors in an endemic region of Australia. Lymphocyte proliferation, activation and cytokine expression to B. pseudomallei antigen were determined in eight donors who were seropositive and in eight age- and sex-matched controls. In North Queensland, 2.5% of the population was seropositive for B. pseudomallei, which is less than half that which was previously described. Of clinical significance was the observation that while 75% of the seropositive individuals had increased lymphocyte proliferation to B. pseudomallei antigens, there were no significant differences observed in lymphocyte activation or production of cytokines. PMID- 19121693 TI - Burkholderia pseudomallei enhances maturation of bone marrow-derived dendritic cells. AB - T-cell activation is essential for protection against Burkholderia pseudomallei infection. Using bone marrow-derived dendritic cells (BMDC) isolated from partially resistant C57BL/6 and susceptible BALB/c mice, the degree of BMDC activation in the presence of B. pseudomallei was investigated. Maturation, cytokine production and internalization of B. pseudomallei by BMDC was assessed in response to infection with a highly virulent and a low-virulent clinical isolate. Maturation was determined by identifying the up-regulation of cell surface markers CD11c and CD86. IL-1beta and IL-12p40 expression were assessed by reverse-transcriptase PCR. The uptake of B. pseudomallei by BMDC was measured using an internalization assay. This study demonstrated that B. pseudomallei isolates stimulate the maturation of BMDC to the same degree regardless of virulence. However, maturation of BMDC was significantly increased in BALB/c mice compared with C57BL/6 mice. Additionally, the uptake of B. pseudomallei by BMDC was significantly greater with the highly virulent isolate compared with the low virulent isolate. Expression of IL-12 and IL-1beta following infection with B. pseudomallei was up-regulated. The differences observed may have implications in the development of an effective immune response to B. pseudomallei. PMID- 19121694 TI - Activation of human monocyte-derived dendritic cells by Burkholderia pseudomallei does not require binding to the C-type lectin DC-SIGN. AB - Dendritic cells (DCs) are essential in regulating adaptive immunity. DC-SIGN (DC specific ICAM-grabbing nonintegrin) is a C-type lectin receptor that is expressed mainly by DCs. Accumulating evidence supports that certain pathogens target DC SIGN to escape host immunity. To investigate a possible role of DC-SIGN in Burkholderia pseudomallei infection, we initially screened its DC-SIGN binding activity by an ELISA method utilizing a DC-SIGN-Fc chimeric protein and found that all of the B. pseudomallei strains tested failed to bind DC-SIGN. However, one strain, the LPS mutant SRM117, which lacks the type II O-polysaccharide expression, actually bound DC-SIGN, in contrast to its wild-type counterpart 1026b (P<0.001). We also found that, although the LPS mutant could readily activate monocyte-derived human DCs, it induced lower levels of IL-12p70 and IL 10 production than its wild-type counterpart (P<0.01). By contrast, the wild-type and the LPS mutants were indistinguishable from one another in terms of T(H)1/T(H)2 differentiation. Altogether, these data suggest that, unlike other certain host pathogen interactions, activation of DCs by B. pseudomallei is not dependent on DC-SIGN. We also found evidence that the LPS mutant that binds DC SIGN has a suppressive effect on DC cytokine production. PMID- 19121695 TI - The effect of different Burkholderia pseudomallei isolates of varying levels of virulence on toll-like-receptor expression. AB - The purpose of this investigation was to ascertain the degree of toll-like receptor (TLR) activation by Burkholderia pseudomallei isolates with varying levels of virulence 2 h post infection. Standard antibiotic protection assays were performed on RAW 264.7 macrophages and peritoneal exudate cells (PEC) challenged with B. pseudomallei. Real-time PCR (RT-PCR) was performed to determine TLR2, TLR4, TLR5 and TLR9 expression. Internalization and killing of bacteria were determined 2h post infection. ELISAs were performed to determine the levels of TNF-alpha from cultured supernatants. Nitrate levels were determined by Griess assays. Up to 2h post infection, B. pseudomallei failed to significantly increase TLR4, TLR5 and TLR9 expression in both cell types. However, TLR2 expression was increased in RAW 264.7 macrophages, irrespective of isolate virulence. The levels of TNF-alpha and nitrate were significantly attenuated in RAW 264.7 macrophages, and no correlation was found between the level of virulence of the infecting strain and TLR expression, bacterial uptake, or killing. The ability of B. pseudomallei to evade detection by macrophages may in part be due to possible signal dampening of TLRs at very early stages of infection. PMID- 19121696 TI - Induction of protective immunity against Burkholderia pseudomallei using attenuated mutants with defects in the intracellular life cycle. AB - Melioidosis is a severe infectious disease caused by the Gram-negative rod Burkholderia pseudomallei. There is currently no vaccine available. We recently generated and characterized several highly attenuated transposon mutants with defects in the intracellular life cycle of B. pseudomallei. In the present study we examined the protective effects of six of these mutants: four harbouring knockouts in genes involved in several biosynthetic pathways (purN(-), purM(-), hisF(-), pabB(-)); a putative lipoate-protein ligase B; and a hypothetical protein. All live mutants conferred protection to some degree against wild-type challenge in susceptible BALB/c mice. Two mutants defective in distinct steps of the purine biosynthetic pathway were selected for further studies. Mutant 30:93 with a defect in the purN gene provided better protection against intraperitoneal challenge than mutant 56:65, which harboured a nonfunctional purM gene. Although mutant 30:93 conferred significant protection against acute fatal disease after intranasal and intraperitoneal challenge with B. pseudomallei, vaccination did not confer protection against chronic forms of melioidosis. Moreover, no protective effect could be seen against intravenous challenge. Further studies are required to analyze the precise nature of the immune response induced by the various live attenuated vaccines with different protective potential. PMID- 19121697 TI - TBK1 does not play a role in the control of in vitro Burkholderia pseudomallei growth. AB - Burkholderia pseudomallei, the causative agent of melioidosis, is an important intracellular pathogen in tropical regions. TANK-binding kinase (TBK1), part of the pathway that induces transcription of Type I interferon genes, has been demonstrated to play an important role in controlling intracellular bacterial infections. To investigate the role of tbk1 in protecting against B. pseudomallei we developed tbk1-deficient cell lines by using shRNA for transient knockdown of the tbk1 gene in HeLa and RAW 264.7 cells. In tbk1-deficient RAW cells, the replication of invasive and non-invasive Escherichia coli was significantly increased at 48 h after infection compared with wild-type cells. The result was confirmed using Brucella melitensis in tbk1-deficient HeLa cells, which demonstrated a >1.5-2.0 log higher bacterial count at 6-48 h after infection compared to wild-type cells. By contrast, the growth of Burkholderia pseudomallei expressing either typical (A2) or atypical (G207) lipopolysaccharide was not significantly different between the tbk1-deficient and control cells. These results suggest that the tbk1 gene and its activation may be able to control invasive E. coli, non-invasive E. coli and B. melitensis growth but may not be able to control B. pseudomallei infection. The role of the tbk1 gene in proinflammatory cytokine induction and bacterial intracellular infection needs further investigation to identify mechanistic differences among the life cycles of various intracellular bacteria. PMID- 19121698 TI - Self-organizing dynamics in protein folding. PMID- 19121699 TI - Toward reliable simulations of protein folding, misfolding and aggregation. PMID- 19121700 TI - Use of protein engineering techniques to elucidate protein folding pathways. PMID- 19121701 TI - Principles of protein misfolding. PMID- 19121702 TI - Protein folding revisited. PMID- 19121703 TI - Minimal models for proteins and RNA from folding to function. PMID- 19121704 TI - The folding pathway of triosephosphate isomerase. PMID- 19121705 TI - Oral health: prevention is key. PMID- 19121706 TI - A favourable (molecular) signal for personalised medicine. PMID- 19121707 TI - America's commitment to global health. PMID- 19121708 TI - Helsinki discords: FDA, ethics, and international drug trials. PMID- 19121710 TI - Paulo Sergio Pinheiro: giving a voice to children. PMID- 19121711 TI - Association between cerebral palsy and erythromycin. PMID- 19121712 TI - The fragmented clinical record: a risk to at-risk children. PMID- 19121713 TI - Association between cerebral palsy and erythromycin. PMID- 19121714 TI - Treatment of extensively drug-resistant tuberculosis. PMID- 19121717 TI - Treatment of extensively drug-resistant tuberculosis. PMID- 19121718 TI - Intermittent preventive treatment with antimalarial drugs. PMID- 19121719 TI - Unsexy tuberculosis. PMID- 19121721 TI - Computerised clinical decision support in rural China. PMID- 19121722 TI - Antibiotic resistance in China--a major future challenge. PMID- 19121723 TI - Unexplained seizures in an infant. PMID- 19121724 TI - Carpal boss: effect of wedge excision depth on third carpometacarpal joint stability. AB - PURPOSE: We hypothesize that carpal-metacarpal (CMC) instability after carpal boss wedge excision is not caused only by damage to the dorsal ligament but mainly depends on the depth of the bony resection. METHODS: To test our hypothesis, this study analyzes the effect of wedge excisions with different depths (0, 15%, 35%, 55% of the third CMC joint) and the effect of different forces (0, 50, 100 N.m) on the stability (measured as the passive flexion) of the third CMC joint using 12 fresh-frozen human cadaver wrists. The passive flexion is defined as the increase in angular motion of the third CMC joint and represents change in stability during flexion of the joint. RESULTS: The results show that the mean passive flexion measured in the wedge excisions of 15% and 35% of the joint did not differ from that of neutral controls. Joints analyzed after a 55% wedge excision showed a significant increase in angular motion (increased passive flexion). This relates to the 50 N.m as well as the 100 N.m loaded test position. CONCLUSIONS: This study shows that a wedge excision of clinically applicable depth of 35% does not create instability during flexion of the third CMC joint when loaded with physiologically relevant forces. Yet an extended and hardly clinically relevant 55% wedge excision results in a change in stability of the joint. To prevent instability when performing a wedge excision for symptomatic carpal boss, care must be taken to avoid excisions that exceed 35% of the third CMC joint. PMID- 19121725 TI - Temporary scaphotrapezoidal joint fixation for adolescent Kienbock's disease. AB - PURPOSE: There are few therapeutic guidelines for adolescent Kienbock's disease. The purpose of our study was to investigate the clinical and radiographic outcomes of temporary scaphotrapezoidal (ST) joint fixation for adolescent Kienbock's disease. METHODS: This was a retrospective review of 6 adolescent patients with Kienbock's disease treated by temporary ST joint fixation. All patients had pain with rest and activity before surgery. The mean patient age at the time of surgery was 14 years, and final follow-up examination was at a mean of 23 months. Under general anesthesia, 2 or 3 K-wires were inserted from the dorsal trapezoid to the scaphoid. The wires were removed at 3 to 6 months. RESULTS: Mean postoperative wrist extension and flexion were increased from 46 degrees and 48 degrees to 68 degrees and 77 degrees , respectively. These improvements were statistically significant compared with preoperative wrist extension and flexion. Grip strength significantly increased from 52% to 86% of the unaffected side. None of the 6 patients had postoperative pain either at rest or with activity. On final follow-up wrist x-ray films, none of the patients had sclerotic change or fragmentation of the lunate, although decreased lunate height remained in all patients. Magnetic resonance imaging revealed improvement to nearly normal intensity of the lunate on T1- and T2-weighted images in all patients. CONCLUSIONS: Both clinical and radiographic outcomes of temporary ST joint fixation for adolescent Kienbock's disease were excellent. We therefore recommend this procedure for the surgical treatment of adolescent Kienbock's disease. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 19121726 TI - Long-term outcomes of trapeziometacarpal arthrodesis in the management of trapeziometacarpal arthritis. AB - PURPOSE: Reported outcomes of trapeziometacarpal (TM) arthrodesis have been contradictory. The purpose of this paper is to review the long-term results of TM arthrodesis for arthritis with respect to clinical outcomes, union, development of adjacent joint arthritis, and complications. METHODS: A retrospective review of TM arthrodeses performed between 1970 and 2003 was undertaken. Among a total of 241 arthrodeses performed, 126 thumbs in 114 patients (79 women, 35 men) treated for osteoarthritis were available for follow-up evaluation. Pre- and postoperative clinical and radiographic data were reviewed. The average age was 57 years (range 32-77). The dominant hand was involved in 76 cases. Supplemental bone graft was used in 90 thumbs. Preoperative appositional (key) pinch, oppositional (tip) pinch, and grip strengths were 3.0 kg, 2.7 kg, and 14 kg, respectively. The average pain score on a scale of 0-10 was 6.6 (range 4-10). The average follow-up was 11.2 years (range 3-28 years). RESULTS: There were 17 nonunions. No correlation existed between the incidence of nonunion and the use of supplemental bone graft. Nine of 17 thumbs had re-operation, including revision arthrodesis (6) and interposition or suspensionplasty (3). The appositional pinch, oppositional pinch, and grip strengths improved to 5.9 kg, 5.4 kg, and 23 kg, respectively (p < .01). The average pain score improved to 0.4 (p < .01). Radiographic progression of scaphotrapeziotrapezoid arthritis occurred in 39 cases; however, only 8 of these were symptomatic. Development of metacarpophalangeal arthritis was noted in 16 thumbs; none have been clinically relevant. CONCLUSIONS: For most patients TM arthrodesis reduces pain, improves function and results in excellent patient satisfaction. Despite the development of metacarpophalangeal and scaphotrapeziotrapezoid joint arthritis, intervention for these joints was rarely warranted. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 19121727 TI - Corrective osteotomy for volarly malunited distal radius fracture. AB - PURPOSE: To retrospectively analyze consecutive cases with opening wedge corrective osteotomy of the volarly malunited distal radius with iliac bone graft, including preoperative and postoperative comparison of symptoms, visual analog scale (VAS), Japanese Society for Surgery of the Hand version of the Disabilities of the Arm, Shoulder, and Hand (DASH-JSSH) questionnaires, radiographic indices, clinical results as evaluated by modified Mayo wrist score, and complications. METHODS: Subjects were 28 patients with volarly malunited distal radius fracture treated by transverse opening wedge osteotomy with oblique iliac bone graft. Preoperative symptoms included wrist deformity, weakness of grip strength, and marked restriction of supination range. Postoperative symptoms, radiographic parameters, clinical results, and complications were analyzed at an average of 25 months of follow-up. We also analyzed the union period of the radius in younger patients (< or =45 years old) and older patients (>45 years old). RESULTS: Mean preoperative visual analog scale was 45, improving significantly to 3 postoperatively. Range of wrist motion improved in all 28 patients, with supination range improving from 16 degrees preoperatively to 80 degrees postoperatively. Mean preoperative DASH-JSSH score was 55, improving to 9 postoperatively. Radiography revealed that volar tilt improved from 32 degrees preoperatively to 10 degrees postoperatively, and radial inclination increased from 17 degrees to 21 degrees . Preoperative ulnar variance of +5.9 mm was corrected to -0.1 mm postoperatively. All 28 wrists demonstrated bony union at the osteotomy site after an average of 52 days (younger patients, 51 days; older patients, 54 days). No complications, significant radiographic correction loss, or nonunion were seen after at least 2 years of follow-up. Modified Mayo wrist score was excellent in 16 patients, good in 10 patients, and fair in 2 patients. CONCLUSIONS: Opening wedge osteotomy for volarly malunited distal radius fracture restored bony configuration of the distal radius, decreased pain, and improved grip strength and range of wrist motion, particularly for forearm supination. No complications or noteworthy correction loss were noted after surgery, even in older patients. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 19121728 TI - A pilot feasibility study for ultrasound evaluation of living human wrist cartilage: site-specific differences in acoustic properties. AB - PURPOSE: The field of cartilage repair has changed dramatically in the past decade but has not answered the question of how to treat an articular cartilage lesion in the wrist. Indeed, the characteristics of wrist articular cartilage, such as cartilage thickness, hardness, and smoothness, have not been clarified. The purpose of this study was to evaluate and quantify the acoustic properties of wrist articular cartilage quantitatively using a new acoustic probe under arthroscopic observation. METHODS: We evaluated 10 consecutive patients (9 men, 1 woman) who were examined or treated arthroscopically. The mean age at evaluation was 27 years. In total, 468 points of wrist articular cartilage were investigated using the ultrasonic probe, and the data were transformed into a wavelet map by wavelet transformation. Two parameters, maximum magnitude and echo duration, which are indices of articular cartilage stiffness and macroscopic surface roughness, respectively, were used to evaluate the acoustic properties of wrist cartilage. RESULTS: The distribution pattern of the acoustic properties was similar to that of previous results for the ankle joint. The mean maximum magnitude and echo duration were 3.41 +/- 1.50 (range, 0.89-7.53) and 1.33 mus +/ 0.30 (range, 0.51-2.17 mus), respectively. For the scaphoid fossa, the maximum magnitude of the radial side was significantly lower than that of the ulnar side, and the echo duration of the radial side was significantly longer than that of the ulnar side. CONCLUSIONS: A new measurement system using an acoustic probe made it possible to perform a quantitative analysis of wrist articular cartilage, similar to the case for knee and ankle articular cartilage. In addition, site specific differences in the acoustic properties of the distal radial cartilage were detected in living human wrist cartilage. PMID- 19121729 TI - Osteochondroma of the capitate causing rupture of the extensor digiti minimi: case report. AB - Osteochondromas usually arise from the metaphyses of long bones. Involvement of the carpal bones is extremely rare. We report a case of an osteochondroma originating from the dorsal capitate in a 51-year-old man. Attritional rupture of the extensor digiti minimi tendon resulted from mechanical irritation as it coursed over the mass. Surgical excision of the mass with tendon transfer of the extensor digiti minimi was subsequently performed. PMID- 19121730 TI - Prevention of recurrent radioulnar heterotopic ossification by combined indomethacin and a dermal/silicone sheet implant: case report. AB - A 27-year-old, right-handed man developed severe radioulnar synostosis at the distal radius 7 months after open reduction and internal fixation of his distal radius fracture. Heterotopic ossification formed at the radius fracture site, requiring excision of the heterotopic bone and plate removal. A bilayer, dermal substitute and silicone sheet was placed between the radius and ulna in the interosseous space to prevent recurrence of the heterotopic ossification. After surgery, the patient was treated with indomethacin 25 mg orally 3 times daily for 6 weeks. At 1 year after surgery, he has retained full pronation and near normal supination. Radiographs demonstrate no new heterotopic bone formation. PMID- 19121731 TI - Digital rewarming patterns after median and ulnar nerve injury. AB - PURPOSE: Posttraumatic cold intolerance (CI) is a frequent and important sequel after peripheral nerve injury. In this study, it is hypothesized that altered rewarming patterns after peripheral nerve injury are related to the degree of posttraumatic CI. This hypothesis is tested by quantitatively comparing rewarming patterns of the digits in controls and in median or ulnar nerve injury patients and by investigating relationships between rewarming patterns, sensory recovery, and CI. METHODS: Twelve median or ulnar nerve injury patients with a follow-up of 4 to 76 months after nerve repair and 13 control subjects had isolated cold stress testing of the hands. Video thermography was used to analyze and compare rewarming patterns of the injured and uninjured digits after cold stress testing. Temperature curves were analyzed by calculating the Q value as an indicator of heat transfer (temperature added during the first 10 minutes after start of active rewarming) and the maximum slope. RESULTS: Test-retest reliability was 0.64 and 0.79, respectively, for the Q value and maximum slope. High Q values and maximum slopes were interpreted as the presence of active rewarming. Patients with return of active rewarming had better sensory recovery and lower Blond McIndoe Cold Intolerance Severity Scale (CISS) scores. Better sensory recovery was correlated with lower CISS scores. CONCLUSIONS: Test-retest reliability of cold stress testing was good, and we found a difference in rewarming patterns between nerve injury patients and controls. The presence of active rewarming in the nerve injury patients was related to sensory recovery and fewer complaints of posttraumatic CI. PMID- 19121732 TI - Shear strain and motion of the subsynovial connective tissue and median nerve during single-digit motion. AB - PURPOSE: The objective of this study was to measure the relative motion of the middle finger flexor digitorum superficialis (FDS) tendon, its adjacent subsynovial connective tissue (SSCT), and the median nerve during single-digit motion within the carpal tunnel in human cadaver specimens and to estimate the relative motions of these structures in different wrist positions. METHODS: Using fluoroscopy during simulated single-digit flexion, we measured the relative motion of the middle finger FDS tendon, SSCT, and median nerve within the carpal tunnel in 12 human cadavers. Measurements were obtained for 3 wrist positions: neutral, 60 degrees flexion, and 60 degrees extension. After testing with an intact carpal tunnel was completed, the flexor retinaculum was cut with a scalpel, and the same testing procedure was repeated for each wrist position. The relative motions of the tendon, SSCT, and median nerve were compared using a shear index, defined as the ratio of the difference in motion along the direction of tendon excursion between 2 tissues divided by tendon excursion, expressed as a percentage. RESULTS: Both tendon-SSCT and tendon-nerve shear index were significantly higher in the 60 degrees of wrist flexion and extension positions than in the neutral position. After division of the flexor retinaculum, the shear index in the 60 degrees wrist extension position remained significantly different from that of the neutral position. CONCLUSIONS: We have found that the relative motion between a tendon and SSCT in the carpal tunnel is maximal at extremes of wrist motion. These positions may predispose the SSCT to shear injury. PMID- 19121733 TI - Partial ulnar nerve transfer to the nerve to the biceps for the treatment of brachial plexopathy in metastatic breast carcinoma: case report. AB - Brachial plexus neuropathy can cause progressive pain and disability in patients with breast cancer. Metastatic spread and radiation injury are the most common causes in these patients. We report a case of partial ulnar nerve transfer to the nerve to the biceps muscle to restore elbow flexion in a patient with combined radiation-induced and metastatic brachial plexopathy. PMID- 19121734 TI - End-to-side neurorrhaphy for median nerve repair after elbow tumor resection: case report. AB - We report a case of a median to ulnar end-to-side neurorrhaphy with epineurectomy but without fascicular donor nerve section. Surgery was performed for the curative treatment of an elbow synovial sarcoma at the level of the elbow that necessitated resection of 10 cm of the median nerve. The patient was followed up for 24 months. After 4 months, a recovery of protective sensation in the territory of the median nerve was noticed and had stabilized by 1 year. PMID- 19121735 TI - Gliding resistance and strength of composite sutures in human flexor digitorum profundus tendon repair: an in vitro biomechanical study. AB - PURPOSE: Although the strength of a tendon repair is clearly important, the friction of the repair is also a relevant consideration. The purpose of this study was to characterize the frictional coefficient, gliding resistance, and breaking strength of suture materials and a suture construct commonly used for flexor tendon repair. METHODS: We measured the friction coefficients of 3-0 braided nylon enclosed in a smooth nylon outer shell (Supramid, S. Jackson, Alexandria, VA), 3-0 braided polyester coated with polybutilate (Ethibond, Ethicon, Somerville, NJ), and 3-0 braided polyester/monofilament polyethylene composite (FiberWire, Arthrex, Naples, FL) sutures. We also measured the gliding resistance, linear breaking strength, and resistance to gapping of zone 2 modified Pennington tendon repairs with the 2 lowest-friction sutures in 20 human cadaveric flexor digitorum profundus tendons. RESULTS: The braided polyester/monofilament polyethylene composite had a significantly lower friction coefficient (0.054) than either the coated polyester (0.076) or nylon (0.130) sutures. The gliding resistances of the repaired tendons with braided/monofilament polyethylene composite suture and those of coated, braided polyester were similar. The strength of the 2 repairs (force to produce a 2 mm gap) and resistance to gap formation were also not significantly different. CONCLUSIONS: Braided polyester composite is a low-friction suture material. However, when this suture was used for tendon repair with a locking suture technique, it did not show a significant effect on the gliding resistance and repair strength compared with the same repair using a coated polyester suture. PMID- 19121736 TI - Biomechanical comparison of modified Kessler and running suture repair in 3 different animal tendons and in human flexor tendons. AB - PURPOSE: To establish the animal flexor tendon that best mimics the biomechanical performance of human flexor tendons. We investigated the biomechanical behavior of core and running sutures in 3 different animal flexor tendons and in human flexor tendons. Additionally, we attempted to help standardize future flexor tendon studies. To that purpose, nearly all variables occurring in the test setup have been highlighted. METHODS: The species selected were pig, calf, sheep, and human. Two groups were formed. In the first group we tested 3-0 core sutures (Ticron; Tyco Healthcare, Vienna, Austria), and in the second group we tested 5-0 running sutures (Ethilon; Ethicon, Vienna, Austria). In each group, 10 tendons of each specimen were tested, which yielded a total of 80 tendons. In each group, the repaired tendons were subjected to 3,000 linear load cycles at a load of 15 N. At the end of this procedure, final gap values were recorded. In the next step, ultimate load-to-failure data were obtained from each specimen. RESULTS: Core sutures behaved similarly in the human, sheep, and pig tendons with respect to ultimate loads. With respect to gap formation, core sutures behaved similarly in the human, sheep, and calf tendons. Deep running sutures behaved similarly in the human, sheep, and pig tendons in terms of ultimate load to failure. CONCLUSIONS: In this study, sheep tendons were found to best mimic the biomechanical behavior of human tendons. Calf tendons seem to be unsuitable. There is a strong need for consistency in biomechanical test setups. PMID- 19121737 TI - Effect of the posterior bundle of the medial collateral ligament on elbow stability. AB - PURPOSE: The role of the posterior bundle of the medial collateral ligament in stability of the elbow remains poorly defined. The purpose of this study was to determine the effect of sectioning the posterior bundle of the medial collateral ligament on the stability of the elbow. METHODS: Varus and valgus gravity-loaded passive motion and simulated active vertical motion were performed on 11 cadaveric arms using an in vitro elbow motion simulator. Varus/valgus angle and internal/external rotation of the ulna with respect to the humerus were recorded using an electromagnetic tracking system in varus, valgus, and vertical orientations. Testing was performed on the intact elbow and after sectioning of the posterior bundle of the medial collateral ligament. RESULTS: With active flexion in the vertical position, the varus/valgus kinematics were unchanged after sectioning of the posterior bundle of the medial collateral ligament. However, in pronation, there was an increase in internal rotation after sectioning of the posterior bundle of the medial collateral ligament compared with that of the intact elbow. This rotational difference was not detected with the forearm in supination. During supinated passive flexion in the varus position, sectioning of the posterior bundle of the medial collateral ligament resulted in increased varus angulation at all flexion angles. In pronation, varus angulation and internal rotation both increased. In supination, sectioning of the posterior bundle of the medial collateral ligament had no effect on maximum varus valgus laxity or maximum internal rotation. However, in pronation, the maximum varus-valgus laxity increased by 3.5 degrees (30%) and maximum internal rotation increased by 1.0 degrees (29%). CONCLUSIONS: These results indicate that isolated sectioning of the posterior bundle of the medial collateral ligament causes a small increase in varus angulation and internal rotation during both passive varus and active vertical flexion. This study suggests that isolated sectioning of the posterior bundle of the medial collateral ligament may not be completely benign and may contribute to varus and rotation instability of the elbow. In patients with insufficiency of the posterior bundle of the medial collateral ligament, appropriate rehabilitation protocols (avoiding forearm pronation and shoulder abduction) should be followed when other injuries permit. PMID- 19121739 TI - Return to work in the setting of upper extremity illness. PMID- 19121738 TI - Identification of biomarkers in Dupuytren's disease by comparative analysis of fibroblasts versus tissue biopsies in disease-specific phenotypes. AB - PURPOSE: Biomarkers are molecular mediators that can serve as indicators of normal biological processes, pathologic processes, and therapeutic interventions. This study aims to identify potential biomarkers in Dupuytren's disease (DD), a fibroproliferative benign tumor with an unknown etiology and high recurrence after surgery. METHODS: Bioinformatic analytical techniques were employed to identify candidate genes that may be differentially expressed in DD, which included gene expression analysis of microarray data and thorough literature searches in genetic linkage and other related biomolecular studies. All DD cases were males with advanced DD (n = 5, 66 years +/- 14). RNA was extracted from biopsies and corresponding cultures of normal fascia (unaffected transverse palmar fascia), palmar nodule and cord from each patient. Real-time reverse transcription-polymerase chain reactions were performed to determine the gene expression levels for disease-related transcripts. RESULTS: The bioinformatic analysis revealed 25 candidate genes, which were further short-listed to 6 genes via functional annotation. The 6 selected candidate genes included: A disintegrin and metalloproteinase domain (ADAM12), aldehyde dehydrogenase 1 family member (ALDH1) A1, Iroquois homeobox protein 6 (IRX6), proteoglycan 4 (PRG4), tenascin C (TNC), and periostin (POSTN). The culturing treatments were shown to have significant impact on the gene expression for ALDH1A1, PRG4, and TNC. In tissue biopsies, significant fold changes were observed for ADAM12, POSTN, and TNC in the cord and/or nodule when compared with that of normal fascia. ADAM12 and POSTN are associated with accelerated or abnormal cell growth, whereas TNC has been associated with fibrotic diseases and cell migration. CONCLUSIONS: This study demonstrated differential gene expression results in DD tissue biopsies compared with that of their corresponding cultures. ADAM12, POSTN, and TNC were identified from the cord and nodule biopsy samples as potential biomarkers in relation to DD development. PMID- 19121740 TI - Digital block anesthesia. PMID- 19121741 TI - Vascularized bone grafting for treatment of Kienbock's disease. AB - Treatment of Kienbock's disease has historically been determined by staging, ulnar variance, and presence or absence of arthritic changes. With the advent of newer techniques of vascularized bone grafting, the status of the cartilage shell of the lunate has become another factor that can influence the procedure performed. The purpose of this article is to describe the technique of the 4th plus 5th extensor compartment artery vascularized bone graft for Kienbock's disease. PMID- 19121742 TI - Lunate revascularization with artery implantation and bone grafting. AB - Treatment of Kienbock's disease encompasses a wide array of options. Proper staging is the first step to successful treatment. Revascularization of the lunate remains a viable option for the treatment of Lichtman stage II or IIIA Kienbock's disease. In this article, we review the indications for and the surgical technique used with a modified Hori procedure (vascular pedicle insertion with structural distal radius graft) for revascularization of the lunate. PMID- 19121743 TI - The influence of ligamentous laxity and gender: implications for hand surgeons. PMID- 19121744 TI - Tissue engineering of bone: a primer for the practicing hand surgeon. PMID- 19121745 TI - Kienbock's disease. AB - Osteonecrosis of the lunate, Kienbock's disease, is a progressive, debilitating disease process that can lead to chronic pain and dysfunction. Despite its recognition almost 100 years ago, the etiology remains unidentified, although mechanical, vascular, and traumatic factors have been implicated. The natural history of this disease is poorly defined, and the radiographic appearance does not always correlate with the clinical findings. Some progress has been made in the identification and an understanding of the progression of the avascular process and its deleterious effects on wrist mechanics. Initial treatment is nonsurgical. Advances in surgical techniques with vascularized pedicled grafts from the distal radius may lead to an improvement in outcomes for patients in the earlier stages of disease, although much more work is needed to determine whether this surgical option represents an improvement over conventional treatment alternatives. Recent reports of long-term outcomes of radial shortening osteotomy for earlier stages of osteonecrosis and proximal row carpectomy for advanced Kienbock's disease reveal that these procedures provide reliable options for the long-term management of this difficult clinical problem. PMID- 19121746 TI - Elbow fractures: distal humerus. AB - A distal humerus fracture can be a debilitating and difficult injury to treat. The anatomy of the distal humerus is highly complex, as it articulates with both the radius and ulna and allows for motion in multiple planes. Furthermore, comminution and osteopenia may render the metaphyseal-diaphyseal junction weak, making adequate stabilization difficult. Various methods of surgical fixation have been described, with bicolumnar plating being the most popular. Controversy over fixation techniques and the introduction of recently developed implants, including precontoured plates and locking plates, have led to renewed focus on biomechanical testing of various fixation constructs. Failure of adequate reconstruction or fixation can be addressed with adjunctive measures such as incorporation of structural bone grafts, external fixation, or, in certain instances, salvage with total elbow arthroplasty. The articular surface can also be injured from a shear force, resulting in fractures of the capitellum and trochlea in the coronal plane. This article presents a review of current literature concerning the diagnosis, classification, treatment, and outcome of distal humerus fractures. PMID- 19121747 TI - Foreword. PMID- 19121748 TI - Preface. PMID- 19121749 TI - Wound complications. AB - This article discusses postoperative wound complications in detail, including host factors that predispose the patient to nonhealing, technical factors in surgery that can reduce the likelihood of infection and dehiscence, and recommendations for postoperative management that can prevent wound healing problems. This discussion includes the treatment of wound complications, ranging from local wound care to various wound coverage options. PMID- 19121750 TI - Revisional peripheral nerve surgery. AB - Peripheral nerve disorders are difficult to manage. In the surgical treatment of patients with peripheral nerve pathology, there are a multitude of factors that may alter the outcome of the patient's recovery and lead to incomplete recovery or possibly worsening of symptoms. The anatomy and function of the peripheral nerve is unique and the evaluation and management of these disorders must be approached in a manner different from musculoskeletal disorders. Many anatomic areas can tolerate scar tissue and adhesions, but in peripheral nerves, loss of the gliding functional and adherence to surrounding soft tissue structures is a common complication from over-zealous dissection and repeat peripheral nerve surgery without modification of technique. The approach to each patient must be thorough and individualized to treat their specific condition, and the surgeon must be aware that at times, the most appropriate treatment for the patient may not be medical but surgical management of the chronic pain condition. PMID- 19121751 TI - Complications and salvage of elective central metatarsal osteotomies. AB - In order to provide proper treatment intervention, the foot and ankle surgeon must develop a further understanding of the pathoanatomy and pathomechanics leading to specific surgical complications of central metatarsal osteotomies. In addition to providing the authors' experiences and potential solutions with regard to these complications, a clear definition of the progression of the complication course is presented. The specific complications of floating toe deformity, metatarsalphalangeal joint stiffness, recurrent metatarsalgia, transfer lesions, malunions, and nonunions are discussed. PMID- 19121753 TI - Revision of failed flatfoot surgery. AB - Revision of failed flatfoot surgery presents a unique and challenging dilemma for the foot and ankle surgeon. Revision surgery is focused on establishing a plantigrade foot with correction of the hindfoot valgus, midfoot abduction, and forefoot varus. Successful reconstruction of failed flatfoot surgery begins with a proper evaluation. No treatment algorithm exists for the management of the malaligned flatfoot. Patient complaints, an understanding of the initial deformity and biomechanical problems, and surgeon experience play a role in correction of failed flatfoot surgery. PMID- 19121752 TI - Revision surgery of the first ray. AB - When dealing with the first ray, and in particular when preparing for hallux abductovalgus surgery, numerous guidelines are available to help decide what procedure is most appropriate for the pathology based on radiographic findings. However, these guidelines are not as black and white as some physicians may think. Numerous factors affect the selection of a procedure, including physician preference, physician comfort with a procedure, experience with a procedure, and surgical skills. This article takes a clinical look at a number of complications related to surgery and trauma of the first ray, and presents a critical discussion of the thought process used to address the complication. PMID- 19121754 TI - Revisional hindfoot arthrodesis. AB - Salvage of a failed hindfoot arthrodesis is an extensive undertaking for the surgeon and patient. With increased morbidity and postoperative convalescence and complications, patients must understand the risk involved in this type of revisional surgery. This article provides a systematic approach to revisional hindfoot arthrodeses, focusing on patient evaluation, surgical technique, and postoperative treatment. PMID- 19121755 TI - Surgical treatment of calcaneal fracture malunions and posttraumatic deformities. AB - The surgical management of calcaneal fractures presents with several obstacles to the treating physician. Many experienced surgeons acknowledge a steep and significant learning curve in the operative management of calcaneal fractures. Nonoperative management of displaced intra-articular calcaneal fractures may result in malunion, thereby affecting the function of the ankle and subtalar joint. Although some calcaneal fractures can be treated conservatively, a majority of them require operative intervention. The goal of this article is to bring some insight into the realm of revisional surgery on residual deformity of the calcaneus after operative intervention and also provide a rationale approach to successfully manage failed surgeries for calcaneal fractures. PMID- 19121756 TI - Talus fractures: surgical principles. AB - Surgical treatment of talus fractures can challenge even the most skilled foot and ankle surgeon. Complicated fracture patterns combined with joint dislocation of variable degrees require accurate assessment, sound understanding of principles of fracture care, and broad command of internal fixation techniques needed for successful surgical care. Elimination of unnecessary soft tissue dissection, a low threshold for surgical reduction, liberal use of malleolar osteotomy to expose body fracture, and detailed attention to fracture reduction and joint alignment are critical to the success of treatment. Even with the best surgical care complications are common and seem to correlate with injury severity and open injuries. PMID- 19121757 TI - Management of complications of open reduction and internal fixation of ankle fractures. AB - The management of complications resulting from the open reduction and internal fixation of ankle fractures is discussed in detail. The initial radiographic findings of the most common postsurgical complications of ankle fracture reduction are briefly discussed, namely lateral, medial, and posterior malleolar malunion or nonunion, syndesmotic widening, degenerative changes, and septic arthritis with or without concomitant osteomyelitis. Emphasis is placed on the management of these complications, with a review of the treatment options proposed in the literature, a detailed discussion of the authors' recommendations, and an inclusion of different case presentations. PMID- 19121758 TI - Revisional Charcot foot and ankle surgery. AB - Charcot neuroarthropathy is often a devastating diabetic foot complication that poses a great risk for limb loss and can have a significant impact on a patient's quality of life in the presence of multiple existing comorbidities. It is a progressive and debilitating condition characterized by joint dislocation, pathologic fracture(s), and extensive destruction of the foot or ankle architecture secondary to dense peripheral neuropathy. This pathologic process can be idiopathic, secondary to acute trauma or previous surgery, or attributable to repetitive "microinjury." Once the Charcot process has been initiated, continued ambulation results in progressive collapse and deformity. Severe deformities can have an impact on the patient's ambulatory status, and when associated with instability, ulceration, or infection, there is greater risk for a major limb amputation. PMID- 19121759 TI - The use of Ilizarov technique as a definitive percutaneous reduction for ankle fractures in patients who have diabetes mellitus and peripheral vascular disease. AB - The patient who has a diagnosis of diabetes mellitus, diabetic peripheral neuropathy, peripheral vascular disease and experiences an unstable ankle fracture presents as a difficult case scenario for the treating physician. In addition, patients who have diabetes mellitus, along with the presence of multiple comorbidities, have been shown to have higher complication rates than patients who do not have diabetes mellitus. This article describes a relatively safe alternative surgical percutaneous technique using external circular ring fixation in the vascularly compromised diabetic patient with an unstable ankle fracture. This novel technique decreases the risk for soft tissue complications in the high-risk diabetic patient and serves as a definitive method of fixation without the need for additional surgery. It allows the patient to have early and full weight bearing when indicated in the postoperative period. PMID- 19121760 TI - Pan-metatarsophalangeal joint arthrodesis for the severe rheumatoid forefoot deformity. AB - The standard approach for correction of severe painful rheumatoid forefoot deformities has involved resection of the metatarsal heads with realignment of the lesser toe deformities and first metatarsophalangeal joint (MTPJ) arthrodesis. Modifications of this procedure may include a pan-metatarsal head resection, including the first metatarsal head, or resection of the lesser metatarsal heads in conjunction with an interpositional arthroplasty of the first MTPJ. The authors describe a novel surgical approach that involves the correction of severe rheumatoid forefoot deformities through a pan-MTPJ arthrodesis. Arthrodesis of all five MTPJs for the surgical treatment of the painful rheumatoid forefoot deformity with chronic plantar callosities and dislocated digits has yet to be reported in the scientific literature. The goal of this article is to provide the treating physician with another alternative and safe surgical approach when dealing with the painful rheumatoid forefoot deformity. PMID- 19121761 TI - Proceedings of the XIVth International Congress on Nutrition and Metabolism in Renal Disease, Marseilles, June 11-15, 2008. Introduction. PMID- 19121762 TI - Mechanisms and treatment of anorexia in end-stage renal disease patients on hemodialysis. AB - Anorexia, defined as the loss of the desire to eat, is relatively common in hemodialysis patients, occurring in one third of such cases. The pathogenesis is essentially unknown. It has been proposed that uremic toxins such as middle molecules, inflammation, altered amino acid pattern, leptin, ghrelin, and neuropeptide Y are involved. Anorexia reduces oral energy and protein intakes, thus contributing to the development of malnutrition and cachexia. Unquestionably, it contributes to poor quality of life. The clinical relevance of anorexia as an independent prognostic factor in hemodialysis is debated. The treatment of this debilitating condition is based on a therapeutic strategy that may include daily dialysis sessions and nutritional counseling. Normalization of plasma branched chain amino acids through branched chain amino acid supplementation may decrease anorexia and improve energy and protein intake. The role of megestrol acetate as an appetite stimulant needs to be validated through adequate randomized trials. Subcutaneous ghrelin administration and melanocortin receptor antagonists appear to be promising therapeutic interventions. PMID- 19121763 TI - Identification of patients with eating disorders: clinical and biochemical signs of appetite loss in dialysis patients. AB - Patients with chronic kidney disease (CKD) frequently experience loss of appetite (anorexia), which increases in severity during the disease progression. The optimal medical assessment of an anorectic patient depends on an appropriate knowledge of the associated signs and symptoms. Some of these signs are diagnostic of anorexia in CKD. The aim of the present review is to provide a general understanding of the consequences of anorexia in the CKD patient, while describing simple assessment methods of appetite loss easy to implement in the clinical setting. Early clinical and physical diagnostic signs associated with appetite loss, as well as the expected reflections in several blood biomarkers, are discussed. Finally, the potential role of sex hormones in modulating the severity of these symptoms is introduced as a platform toward the understanding of sex hormone action in regulating/treating uremic anorexia. PMID- 19121764 TI - New measurements of energy expenditure and physical activity in chronic kidney disease. AB - The accurate estimation of total daily energy expenditure (TEE) in chronic kidney patients is essential to allow the provision of nutritional requirements; however, it remains a challenge to collect actual physical activity and resting energy expenditure in maintenance dialysis patients. The direct measurement of TEE by direct calorimetry or doubly labeled water cannot be used easily so that, in clinical practice, TEE is usually estimated from resting energy expenditure and physical activity. Prediction equations may also be used to estimate resting energy expenditure; however, their use has been poorly documented in dialysis patients. Recently, a new system called SenseWear Armband (BodyMedia, Pittsburgh, PA) was developed to assess TEE, but so far no data have been published in chronic kidney disease patients. The aim of this review is to describe new measurements of energy expenditure and physical activity in chronic kidney disease patients. PMID- 19121765 TI - Insulin resistance in chronic uremia. AB - Insulin resistance often characterizes chronic uremia, and is associated with enhanced morbidity and mortality, because it may contribute to protein-energy wasting (in turn, an independent predictor of reduced survival), atherosclerosis, and cardiovascular death. Causes of insulin resistance in chronic uremia are complex and multifactorial. Obesity is emerging as an independent risk factor for chronic kidney disease, and an expected rise in number of obese uremic patients because of the ongoing worldwide obesity epidemic is likely to increase the prevalence of insulin resistance in chronic uremia in the near future. Similar to the general population, reported associations between obesity and insulin resistance in chronic uremia support a role of adipose tissue and altered adipokine profiles in insulin resistance in obese chronic kidney disease patients. Hormonal imbalances, chronic acidosis, and systemic inflammation and oxidative stress are uremia-associated relevant causes of insulin resistance in nonobese individuals. A further understanding of the causes of insulin resistance in chronic uremia represents a potential important tool in the design of more effective therapeutic strategies to reduce uremia-associated morbidity and mortality. PMID- 19121766 TI - Clinical implications of elevated asymmetric dimethylarginine in chronic kidney disease and end-stage renal disease. AB - Establishing solutes that accumulate in body fluids in renal insufficiency as authentic uremic toxins is a process demanding in vitro and in vivo animal experiments as well as observational and interventional studies in humans. Asymmetric dimethylarginine (ADMA) is listed among the few uremic solutes close to the inference criteria for causal involvement into the pathogenesis of the uremic syndrome. In healthy subjects, the intravenous administration of this methylarginine, such as to bring circulating ADMA to supraphysiologic concentration, raises peripheral vascular resistances and arterial pressure. In uremic patients, ADMA is strongly associated with carotid atherosclerosis and with overall and cardiovascular mortality. Yet we still need the decisive proof that this substance is causally involved in the high risk of death and cardiovascular complications of this population-that is, an intervention study proving that reducing ADMA levels translates into better clinical outcomes. PMID- 19121767 TI - Indoxyl sulfate promotes proliferation of human aortic smooth muscle cells by inducing oxidative stress. AB - OBJECTIVES: Cardiovascular disease is a major cause of mortality in chronic kidney disease patients. We have recently demonstrated that indoxyl sulfate (IS), a uremic toxin, induced aortic calcification and aortic wall thickening in hypertensive rats. This study aimed to determine if IS promotes proliferation of human aortic smooth muscle cells (HASMCs) and if antioxidants inhibit the IS induced cell proliferation. METHODS: We examined the effect of IS at different concentration from 50 to 500 mumol/L on cell proliferation of HASMCs by using 4 [3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST 1) assay. Further, the effect of antioxidants such as vitamin E, vitamin C, and N acetylcysteine on the IS-induced proliferation of HASMCs was determined. RESULTS: IS significantly promoted the proliferation of HASMCs concentration-dependently. The antioxidants significantly inhibited the IS-induced proliferation of HASMCs. CONCLUSION: IS promotes proliferation of HASMCs by inducing oxidative stress. PMID- 19121768 TI - Burnt-out diabetes: impact of chronic kidney disease progression on the natural course of diabetes mellitus. AB - Many individuals with diabetic nephropathy, the leading cause of chronic kidney disease (CKD) in the United States, progress to stage 5 of CKD and undergo maintenance dialysis treatment. Recent data indicate that in up to one third of diabetic dialysis patients with a presumptive diagnosis of diabetic nephropathy, glycemic control improves spontaneously with the progression of CKD, loss of residual renal function, and the initiation of dialysis therapy, leading to normal-to-low hemoglobin A1c (<6%) and glucose levels, requiring cessation of insulin or other anti-diabetic medications. Potential contributors to this so called "burnt-out diabetes" include decreased renal and hepatic insulin clearance, a decline in renal gluconeogenesis, deficient catecholamine release, diminished food intake (because of anorexia or diabetic gastroparesis), protein energy wasting (with resultant loss of weight and body fat), and the hypoglycemic effects of dialysis treatment. Although the concept of "burnt-out diabetes" appears in sharp contradistinction to the natural history of diabetes mellitus, studying this condition and its potential causes and consequences, including the role of genetic factors, may lead to a better understanding of the pathophysiology of metabolic syndrome and diabetes mellitus in the CKD population and in many other individuals with chronic disease states associated with wasting syndrome that can confound the natural history of diabetes. PMID- 19121769 TI - Hypovitaminosis D in chronic kidney disease patients: prevalence and treatment. AB - In the past decade, there has been accumulating evidence indicating that vitamin D may provide great health benefits. Indeed, it has been shown that vitamin D deficiency not only causes bone and mineral disturbances but also may increase the risk of many common chronic diseases. Since very few foods contain vitamin D, humans depend on sun exposure to satisfy their requirements for vitamin D. However, factors related to the location, climate, aging, skin pigmentation, and use of barriers to protect from the sun light contribute to limit the synthesis of vitamin D in the skin. Vitamin D deficiency is highly prevalent in the general population and even more common in patients with chronic kidney disease (CKD). Certain conditions associated with CKD such as protein losses and the decreased food intake can predispose these patients to hypovitaminosis D. The interest in the nutritional status of vitamin D of CKD patients has been renewed after the recognition that low circulating levels of 25 hydroxyvitamin D, and not only of the vitamin D active form (1,25 dihydroxyvitamin D), can contribute to the development of secondary hyperparathyroidism. Therefore, vitamin D supplementation with either ergocalciferol or cholicalciferol has been recommended to restore and maintain adequate vitamin D status in CKD. PMID- 19121770 TI - Bone imaging and chronic kidney disease: will high-resolution peripheral tomography improve bone evaluation and therapeutic management? AB - Bone damage because of chronic kidney disease (CKD) represents a daily challenge for nephrologists. The impact of CKD on bone health may be immediate (serum phosphocalcic disturbances) or delayed (bone fractures and vascular calcifications). Histomorphometry remains the gold standard to evaluate bone, but it is rarely performed in clinical practice. Areal measurement of bone mineral density by dual x-ray absorptiometry is routinely performed to evaluate bone mass. However, this technique presents some limitations. In 2000, the United States National Institutes of Health defined new "quality" criteria for the diagnosis of osteoporosis in addition to decreased bone mass. Bone strength actually integrates two concepts: bone quantity and bone quality (i.e., microarchitectural organization, bone turnover, bone material properties such as mineralization, collagen traits, and microdamage) that cannot be evaluated by dual x-ray absorptiometry. New three-dimensional, noninvasive bone-imaging techniques have thus been developed, e.g., high-resolution peripheral quantitative computed tomography. High-resolution peripheral quantitative computed tomography allows evaluation of both volumetric density and microarchitecture in different compartments of bone, at the distal radius and tibia. High-resolution peripheral quantitative computed tomography may be useful in predicting fractures and assessing bone preventive or therapeutic strategies in CKD patients. It should be evaluated in long-term, longitudinal follow-ups. PMID- 19121771 TI - Klotho gene, phosphocalcic metabolism, and survival in dialysis. AB - The discovery that two recently identified molecules, klotho and fibroblast growth factor 23 (FGF23), played an important role in calcium, phosphate, and vitamin D metabolism has transformed our traditional physiological view in which bone and mineral homeostasis was mainly regulated by parathyroid hormone, vitamin D, and calcitonin, according to mineral body needs. FGF23 is a 251-amino acid secreted protein produced by osteoblasts and osteocytes in bone following the stimulation by phosphate and vitamin D or the inhibition by dentin matrix protein 1. Originally isolated from tumoral cells of patients with tumor-induced osteomalacia and hypophosphatemia, FGF23 inhibits phosphate reabsorption in renal proximal tubular cells and 1alpha-hydroxylase activity, resulting in decreased synthesis of calcitriol. To exert these actions, FGF23 requires the conversion, by klotho, of the canonical FGF receptor 1 (IIIc) in a specific high affinity FGF23 receptor. On the other hand, klotho is a putative antiaging gene identified in 1997 when a particular mouse strain, created by random insertion mutagenesis, was found to be short-lived and displayed premature atherosclerosis, osteopenia, skin atrophy, pulmonary emphysema, hyperphosphatemia, hypercalcemia, and high serum calcitriol levels. The gene of klotho encodes a 1012-amino acid cell surface protein with a short cytoplasmic tail and an extracellular domain that consists in tandem duplicated copies of a beta-glucuronidase-like sequence, which can be released into the circulation as soluble forms after being cleaved by metalloproteinases such as ADAM10 and ADAM17. By modulating FGF23 action, klotho regulates urinary phosphate excretion and calcitriol synthesis. By virtue of its beta-glucuronidase activity, klotho deglycosylates the calcium channel TRPV5 (transient receptor potential vallinoid-5) and regulates urinary calcium excretion. klotho also binds to Na(+),K(+)-ATPase in parathyroid cells and regulates calcium-stimulated PTH secretion. Finally, klotho extends life span via several mechanisms, including the reduction of calcitriol synthesis, serum calcium, and phosphorus levels; the induction of insulin resistance; and by increasing the resistance to oxidative stress. PMID- 19121772 TI - Dietary sodium intake and arterial blood pressure. AB - We sought to summarize major recent studies in the field of dietary sodium intake and arterial blood pressure, and discuss the following trials. INTERSALT: Sodium intake correlates with the rise in blood pressure with age, but not with the prevalence of hypertension. The population study identified a minimal impact of sodium intake on blood pressure (0.9 mm Hg/10 mmol difference in salt intake). DASH: This diet induced significant reductions in blood pressure compared with the control diet. Further decreases were observed with DASH and a 50 mmol/day sodium intake. VANGUARD: Blood pressure was inversely related to urinary potassium, calcium and magnesium but not to sodium excretion. TONE: Cardiovascular events were highest in the usual care group (83%) and lowest in the sodium reduction-plus-weight loss group (56%). META-ANALYSIS: A systematic review of 11 long-term controlled randomized trials reported a small decrease (1.1 mm Hg) in median systolic but not diastolic blood pressure with a reduced dietary sodium intake. In conclusion, (1) sodium restriction in hypertensive patients reduces blood pressure, and (2) the long-term impact of reduced salt intake on blood pressure, mortality, and morbidity remains to be defined. PMID- 19121773 TI - Role of sodium intake in the progression of chronic kidney disease. AB - The relation of salt to hypertension and kidney disease had been well known at the turn of the last century, but the importance of salt has been grossly neglected more recently. There is a close link between salt intake and hypertension, as well as partially blood pressure-independent target organ damage including renal disease. In the general population, high salt intake is associated with hypertension and cardiovascular events. Salt loading also increases albuminuria in individuals without primary renal disease and raises excretion of albumin and protein in patients with renal disease. It aggravates proteinuria and glomerulosclerosis and accelerates progression in most animal models of renal damage. The effect of salt restriction cannot be reproduced by treatment with diuretics. Inappropriate increase of intrarenal angiotensin II and increased reactive oxygen species are the major culprits responsible for salt related renal damage. PMID- 19121774 TI - Salt restriction and practical aspects to improve compliance. AB - Salt as a commodity to preserve and flavor food and to treat ailments has been in existence for hundreds of years. Sodium (salt) restriction to treat hypertension and fluid retention has been practiced for several decades. More recently, evidence-based best practice guidelines have been developed and published on websites to guide medical and healthcare practitioners to treat patients with hypertension, heart disease, chronic kidney disease, and other sodium-sensitive conditions. As the consequences of hypertension affect many people around the world, the World Health Organization is involved in helping to prevent the development of hypertensive risk factors by advocating a global reduction in salt and a healthy lifestyle. Many governments have or are in the process of adapting these recommendations for local implementation. This paper provides information about existing guidelines for healthy individuals and patients with chronic kidney disease and practical aspects to achieve a lifelong change in dietary habits. PMID- 19121775 TI - Salivary phosphorus and phosphate content of beverages: implications for the treatment of uremic hyperphosphatemia. AB - BACKGROUND: Hyperphosphatemia provides relevant and dangerous evidence of end stage renal disease (ESRD) in patients undergoing periodic hemodialysis. The relationship between hyperphosphatemia and cardiovascular calcification, with the consequences of high morbidity and mortality after cardiovascular events, is well defined. Hyperphosphatemia is treated by dietary limitation of phosphorus ingestion and by phosphate binders, but only half of ESRD patients fall within the range of K/DOQI guidelines. OBJECTIVE AND METHODS: We summarize the results of our studies on salivary phosphate secretion in hemodialysis (HD) and chronic kidney disease (CKD) patients, and on the habit of HD patients to drink beverages with a high or low phosphate content. We also examine the correlation between hyperphosphoremia and the phosphate content of common beverages consumed by HD patients. RESULTS AND CONCLUSIONS: Higher levels of salivary phosphate secretion were found in HD and in CKD patients, along with a relationship between serum phosphorus levels and a high phosphate content of beverages in HD patients. PMID- 19121776 TI - Lipid and lipoprotein metabolism in chronic kidney disease. AB - The risk of cardiovascular events and mortality increases as renal function declines although the relative risk of mortality contributed by the standard Framingham risk factors are altered or replaced. Low-density lipoprotein (LDL) cholesterol does not predict mortality but low high-density lipoprotein (HDL) cholesterol and triglycerides remain risk factors. The lipoproteins within each class are shifted to smaller, more dense isoforms. The accumulation of apolipoprotein B-containing lipoproteins, including lipoprotein(a) results primarily from decreased clearance rather than from increased synthesis. Lipoprotein(a) levels are also associated with cardiovascular outcome among dialysis patients. Decreased clearance of very low-density lipoprotein and intermediate-density lipoprotein is a result of decreased lipoprotein lipase, structural alterations in the lipoproteins rendering them poorer substrates, and a decrease in receptor number for these proteins. HDL levels are decreased as a result of an increased fractional catabolic rate both among obese patients with normal renal function and among dialysis patients, but the mechanisms responsible for increased HDL fractional catabolic rate may differ. In patients with advanced kidney disease, HDL fails to mature normally as a result of decreased lecithin cholesterol ester transfer protein, leaving cholesterol ester-poor, triglyceride rich HDL(3) and pre-beta HDL. HDL in patients with chronic kidney disease is a less effective antioxidative agent than is HDL from normal subjects because of a decrease in paroxonase activity, allowing the accumulation of oxidized LDL. PMID- 19121778 TI - Does the uremic milieu affect the epigenotype? AB - Epigenetics is a discipline that for many years has languished in the shadow of its genomic big brother. Because our understanding of the role played by epigenetics in chronic kidney disease remains in its infancy, further studies are needed to understand the associations, for instance, of aberrant DNA methylation in relation to uremic dysmetabolism, and its presumably very complex interactions in the development of premature uremic vascular disease. Further research is also needed to study the association between aberrant global DNA-methylation, gene level methylation status, and the silencing (or activation) of candidate genes associated with atherosclerosis. Insofar as it seems possible to manipulate the epigenome, the effects of various epigenetic-targeted and pathway-targeted therapeutic approaches on unbalanced DNA methylation, gene silencing, and vascular health and outcomes should be explored further in uremia. PMID- 19121777 TI - Vascular calcification in chronic kidney disease: pathogenesis and clinical implications. AB - Cardiovascular disease is the primary cause of morbidity and mortality in chronic kidney disease (CKD). Nontraditional uremia-related risk factors as well as traditional risk factors may contribute to the unique features of cardiovascular disease in patients with CKD. Vascular calcification is a prominent feature of arterial disease in CKD and may have an impact on cardiovascular mortality through modulating both arteriosclerosis (arterial stiffening) and atherosclerosis. There are two pathophysiological processes involved in the development of vascular calcification: apoptosis and phenotypic transition to chondrocytes or osteoblasts (chodro/osteogenic differentiation). In CKD, abnormal mineral metabolism, predominantly hyperphosphatemia and hypercalcemia, facilitates progression of vascular calcification in association with functional disturbances of its inhibitory molecules (inhibitors of vascular calcification) such as pyrophosphate, matrix Gla protein, fetuin-A, and osteoprotegerin. PMID- 19121779 TI - Nutrition and muscle catabolism in maintenance hemodialysis: does feeding make muscle cells selective self-eaters? AB - Efforts to build muscle by increased protein feeding in hemodialysis patients have been thwarted by parallel increases in both muscle protein synthesis and degradation. The evidence suggests that muscle cells replace older proteins in response to feeding rather than using new proteins to drive muscle cell hypertrophy. This review presents the hypothesis that protein feeding provides an opportunity for muscle to accelerate proteolysis of proteins that have been damaged by oxidation, nitrosylation, and/or glycosylation and to replace damaged mitochondria that contribute to oxidative stress. Increases in proteolysis with feeding are driven by insulin resistance and the increased oxidative stress of mitochondrial respiration. Oxidized proteins and organelles are excellent substrates for degradation by the proteasome, macroautophagy, and chaperone mediated autophagy: these systems of proteolysis seem to be activated by oxydatiative stress. Replacement of oxidized and other damaged proteins may be a benefit of protein feeding in hemodialysis, but alternative strategies, including exercise, will be required to build muscle. PMID- 19121780 TI - Hemodialysis and nutritional status in children: malnutrition and cachexia. AB - Malnutrition is a common state in chronic hemodialyzed children. More than malnutrition, which infers that dietary supplementation would be curative, cachexia, which implies loss of protein stores, are combined factors of impaired linear growth and reduced muscle mass in uremic patients. Adequate diet to prevent malnutrition is of major importance. But to avoid cachexia in children on chronic hemodialysis, the management of acidosis, inflammation, abnormal metabolic rate, and endocrine disturbances should not, be forgotten. Daily hemodialysis regimen using convective flow therapy and ultrapure dialysate, i.e., on line hemodiafiltration together with growth hormone therapy, appears as a hopeful strategy for the chronic dialyzed child to achieve catch-up growth, a parameter of optimal nutrition. PMID- 19121781 TI - Nutritional consequences of renal transplantation. AB - Successful kidney transplantation leads to restoration of renal function. Some metabolic disorders from chronic renal failure may persist and new metabolic abnormalities can develop (obesity, diabetes, hypertension, bone disease, and anemia). Additionally, influence of immunosuppressive drugs (corticosteroids, cyclosporine A, tacrolimus, and rapamycin) may aggravate the course of diabetes, hypertension, and dyslipidemia. Nutritional management of renal transplantation is divided into the pretransplant period, transplant surgery, and early and late posttransplant period. Patients in the pretransplant period in dialysis treatment may develop protein-energy malnutrition and negative nitrogen balance, with loss of lean body mass and fat deposits. Nutritional management in the early posttransplant period with a functioning kidney graft necessitates fluid and electrolyte balance control with protein intake of 1,2/kg BW/day and 30-35 kcal/kg BW/day. In a nonfunctioning kidney graft, dialysis treatment continues and the therapeutic dose of immunosuppressive drugs must be reduced. The principal objective in the late posttransplant period is the maintenance of optimal nutritional status. Nutrition is important in managing obesity, insulin resistance, diabetes, hyperlipidemia, and hypertension. Other posttransplant conditions for which diet and/or nutritional supplements may be beneficial include hypomagnesemia, hypophosphatemia, hyperuricemia, hyperkalemia, hyperhomocysteinemia, chronic renal allograft failure, renal anemia, and renal bone disease. PMID- 19121782 TI - Development of evidence-based guidelines for the nutritional management of adult kidney transplant recipients. AB - OBJECTIVE: This article documents the development of evidence-based guidelines for the nutritional management of adult kidney transplant recipients. Dietary interventions play an important role in preventing and managing common post transplant health problems, such as cardiovascular disease and diabetes. However, there are currently no comprehensive, evidence-based guidelines for the nutritional management of kidney transplant recipients. METHODS AND RESULTS: Thirteen guideline topics were identified, including obesity, diabetes, dyslipidemia, and bone disease, following broad consultation with clinicians and transplant recipients in Australia and New Zealand. A systematic review of the scientific literature was undertaken, the protocol for which is published in the Cochrane Library. The evidence was graded and synthesized, and evidence-based recommendations formulated consistent with National Health and Medical Research Council of Australia standards. A total of 119 scientific papers were assessed. CONCLUSION: There was no level I or II evidence to support any guideline; however, there was sufficient level III and IV evidence to support Grade C and D recommendations for six guideline topics. Experts from 18 transplant units in Australia and New Zealand were consulted to generate consensus-based recommendations for the remaining seven topics, using the Delphi method. Using evidence from a comprehensive literature search and expert opinion, guidelines that represent current best practice have been produced. These guidelines have been evaluated in transplant units throughout Australia and New Zealand and have been submitted to the Dietitians Association of Australia (DAA) and Caring for Australasians with Renal Impairment (CARI) for endorsement. PMID- 19121783 TI - Metabolic syndrome after kidney transplantation. AB - Metabolic syndrome (MS) can be linked to the progression of chronic renal transplant dysfunction (CRTD). Hyperfiltration in transplanted patients is a further risk factor for MS and for the progression of CRTD. Many studies show in kidney-transplanted subjects a prevalence of MS at 60% after 6 years posttransplantation. We studied 182 patients (126 men and 56 women) with functioning renal transplant in Messina and Rovigo Renal Units. In our patients we saw at 6 years postkidney transplant in men a prevalence of MS at 20% (NCEPATPIII criteria) or 30% (Rovigo-Messina more strict criteria), and women at 6 years posttransplantation a prevalence of MS at 25% (NCEPATPIII criteria) or 50% (Rovigo-Messina criteria). In our patients, the period of graft function was over 15 years. This result is due to continued follow-up of metabolic parameters and to moderate protein (0.8-0.9 g/kg/day), low lipid, and low caloric intake. These strategies permit the avoidance of hyperfiltration, the reduction of hyperlipidemia and obesity. Special attention is due to women 50-65 years old. In these patients the prevalence of metabolic syndrome 6 years posttransplant is very high (60%). Postmenopausal syndrome is an additional risk factor for MS, atherosclerosis, and the progression of CRTD. PMID- 19121784 TI - Nutritional and metabolic issues in solid organ transplantation: targets for future research. AB - Cardiovascular morbidity and mortality complicates the course of a significant proportion of renal transplant recipients and is increasingly prevalent among recipients of other solid organ transplants, such as heart or liver transplant patients. A posttransplant metabolic syndrome comprised of hypertension, dyslipidemia, increased fat mass/obesity, and glucose intolerance, combined with other metabolic side effects derived from glucocorticoid and calcineurin inhibitor immunosuppression, attenuates allograft and patient survival. After the early posttransplant years, infection and rejection are the major risks that recipients face, whereas metabolic and cardiovascular disease become the most serious long term risk factors impacting patient survival. While significant advances in immunosuppressive therapy have prolonged the allograft and patient survival in solid organ transplant recipients, little has been done in the way of controlled interventional trials utilizing nutritional, dietary, or biobehavioral modification, especially when combined with drug treatment to reduce the effects of the posttransplant metabolic syndrome. In addition to cardiovascular morbidity, metabolic bone disease, osteopenia, and impaired growth in children pose significant challenges in posttransplant management. In this review, the data from some of the known observational dietary trials in solid organ transplant recipients and prior evidence obtained from studies in chronic kidney disease and the general population is considered in formulating new targets for future research to deal with this ever-increasing population of high risk patients. PMID- 19121785 TI - Research in renal nutrition--how to get started. AB - Dietitians, general and renal, have long struggled with the tasks of conducting and publishing nutrition-related research. When surveyed, dietitians indicated that the reasons they are not participating in research, and thus writing and submitting manuscripts for publication, are that (1) they do not have sufficient support from administration and (2) they lack available time. Removing barriers to research participation will alter the perceptions, attitudes, and knowledge of dietitians, thus enabling current practitioners to discover ways in which they can overcome their individual obstacles and participate in research. Increased support and knowledge must be provided to renal dietitians while simultaneously limiting the stress that may occur while assuming these additional responsibilities. Participating in renal nutrition research will result in renal dietitians generating nutrition data on which to base practice, increase their job satisfaction, and increase the level of financial reimbursement for nutrition services. PMID- 19121786 TI - The atrial electrograms and the activation sequence in the CS catheter are identical to the preceding sinus beat. PMID- 19121788 TI - electrical impedance monitoring has only limited effectiveness in preventing adverse clinical events, specifically inappropriate shocks, resulting from fracture of ICD leads. PMID- 19121790 TI - The Sprint Fidelis implantable cardioverter-defibrillator (ICD) leads (Medtronic, Inc., Minneapolis, MN, USA) were removed from the market in October 2007 because of the rate of pace-sense conductor fractures. PMID- 19121792 TI - The Medtronic Lead Integrity Alert (LIA) significantly increases the likelihood of detecting a potential defibrillation lead fracture that could result in inappropriate therapy. PMID- 19121794 TI - Value of implantable cardioverter-defibrillators in managing atrial fibrillation. PMID- 19121795 TI - Relationship between the spectral characteristics of atrial fibrillation and atrial tachycardias that occur after catheter ablation of atrial fibrillation. AB - BACKGROUND: During catheter ablation of complex fractionated atrial electrograms, persistent atrial fibrillation (AF) may convert to an atrial tachycardia (AT). OBJECTIVE: The purpose of this study was to investigate the possible mechanisms of AT by examining the spectral and electrophysiologic characteristics of AF and ATs that occur after catheter ablation of AF. METHODS: The subjects of this study were 33 consecutive patients with persistent AF who had conversion of AF to AT during ablation of AF (group I) and 20 consecutive patients who underwent ablation of persistent AT that developed more than 1 month after AF ablation (group II). Spectral analysis of the coronary sinus (CS) electrograms and lead V(1) was performed during AF at baseline, before conversion, and during AT. The spatial relationship between the AT mechanism and ablation sites was examined. RESULTS: A spectral component with a frequency that matched the frequency of AT was present in the baseline periodogram of AF more often in group I (52%) than in group II (20%, P = .02). Ablation resulted in a decrease in the dominant frequency of AF but not in the frequency of the spectral component that matched the AT. There was a significant direct relationship between the baseline dominant frequency of AF and the frequency of AT in the CS in group I (r = 0.76, P <.0001) but not in group II (r = 0.38, P = .09). ATs were macroreentrant in 64% and 60% of patients in groups I and II, respectively (P = .8). The AT site was more likely to be distant (>1 cm) from AF ablation sites in group I (70%) than in group II (35%, P = .007). CONCLUSION: The findings of this study suggest that ATs observed during ablation of AF often may be drivers of AF that become manifest after elimination of higher-frequency sources and fibrillatory conduction. PMID- 19121796 TI - New methods for estimating local electrical activation rate during atrial fibrillation. AB - BACKGROUND: Dominant frequency (DF) analysis is sometimes limited in its characterization of atrial fibrillation (AF), thus better methodology is needed. OBJECTIVE: To develop methods for analyzing the AF signal without transformation to frequency space, and to compare the methods to DF analysis using both real and simulated data. METHOD: Electrograms were obtained in 11 patients with paroxysmal or persistent AF. Atrial rate (AR) was estimated retrospectively from 8.4-s intervals acquired at 8 sites, using DF and 2 new methods: (1) Signals were converted into merged deflections (time complexes [TC]). Mean peak-to-peak cycle length was used to estimate AR. (2) The ensemble average (EA) was calculated with window size (w) ranging from 50 to 500 ms (frequency f = 2 to 20 Hz). The dominant peak in the root-mean-square power spectrum of EA versus w was used to estimate AR. Methods were compared using patient data, and in simulations using a Hanning pulse (2-mV peak, 50-ms duration, 120-ms period) with additive random noise, amplitude changes, and phase shift. Data from 10 atrial flutter patients were used to validate rate estimation under static conditions. RESULTS: From pooled AF data (N = 88 sites), mean atrial rate was 5.71 Hz (EA), 5.96 Hz (TC), and 5.72 Hz (DF). The mean absolute rate difference was 0.65 Hz (EA-TC), 0.58 Hz (EA-DF), and 0.50 Hz (TC-DF). In simulations, DF, TC, and EA, respectively, correctly predicted rate with additive random noise up to 0.13 mV, 1.4 mV, and 15.2 mV, random phase shift up to 46.3 ms, 28.8 ms, and 51.3 ms, and random amplitude variation ranging from 0 to 4 mV, 1.3 to 2.7 mV, and 0 to 4 mV. All methods accurately estimated rate during atrial flutter. CONCLUSIONS: Accurate rate estimation depends on signal periodicity and is influenced by random noise. The EA provides an accurate rate estimate of the periodic component of AF even at high noise levels. PMID- 19121797 TI - Real-time dominant frequency mapping and ablation of dominant frequency sites in atrial fibrillation with left-to-right frequency gradients predicts long-term maintenance of sinus rhythm. AB - BACKGROUND: Spectral analysis identifies localized sites of high-frequency activity during atrial fibrillation (AF). OBJECTIVE: This study sought to determine the effectiveness of using real-time dominant frequency (DF) mapping for radiofrequency ablation of maximal DF (DFmax) sites and elimination of left to-right frequency gradients in the long-term maintenance of sinus rhythm (SR) in AF patients. METHODS: DF mapping was performed in 50 patients during ongoing AF (32 paroxysmal, 18 persistent), acquiring a mean of 117 +/- 38 points. Ablation was performed targeting DFmax sites, followed by circumferential pulmonary vein isolation. RESULTS: Ablation significantly reduced DFs (Hz) in the LA (7.9 +/- 1.4 vs. 5.7 +/- 1.3, P <.001), coronary sinus (CS) (5.7 +/- 1.1 vs. 5.3 +/- 1.2, P = .006), and RA (6.3 +/- 1.4 vs. 5.4 +/- 1.3, P <.001) abolishing baseline left to-right atrial DF gradient (1.7 +/- 1.7 vs. 0.2 +/- 0.9; P <.001). Only a significant reduction in DFs in all chambers with a loss of the left-to-right atrial gradient after ablation was associated with a higher probability of long term SR maintenance in both paroxysmal and persistent AF patients. After a mean follow-up of 9.3 +/- 5.4 months, 88% of paroxysmal and 56% of persistent AF patients were free of AF (P = .02). Ablation of DFmax sites was associated with a higher probability of remaining both free of arrhythmias (78% vs. 20%; P = .001) and free of AF (88% vs. 30%; P <.001). CONCLUSION: Radiofrequency ablation leading to elimination of LA-to-RA frequency gradients predicts long-term SR maintenance in AF patients. PMID- 19121798 TI - Frequency mapping: hype or hope? PMID- 19121799 TI - Idiopathic focal epicardial ventricular tachycardia originating from the crux of the heart. AB - BACKGROUND: Idiopathic ventricular tachycardia (VT) can arise from the epicardium, usually near the summit of the left ventricle (LV). OBJECTIVE: The purpose of this study was to describe a distinct syndrome of epicardial VT that arises from the crux of the heart. METHODS: Among 340 patients with idiopathic VT referred for ablation, four were identified with VT that was mapped to the epicardium at the crux. RESULTS: VT was sustained in all patients and was associated with syncope or presyncope in three. Rapid VT (mean cycle length 264 ms) was induced with programmed stimulation or burst pacing from the ventricle but required isoproterenol infusion in three. ECG during VT demonstrated a left superior axis QRS morphology with a precordial maximal deflection index >or=0.55 in all patients (mean 0.61). Intracardiac mapping revealed earliest activation in the middle cardiac vein or proximal coronary sinus at the crux in all patients. Irrigated radiofrequency ablation in the middle cardiac vein or proximal coronary sinus was attempted in all patients and successfully abolished VT in one. Percutaneous epicardial radiofrequency ablation was attempted in 2 of 3 remaining patients and successfully abolished VT in both. Simultaneous coronary angiography demonstrated the site of earliest activation within 5 to 10 mm of the proximal posterior descending coronary artery, with no acute narrowing of that artery following ablation. CONCLUSION: Idiopathic VT may arise by a focal mechanism from the epicardium at the crux in close proximity to the posterior descending coronary artery. This syndrome can result in rapid, catecholamine-sensitive VT and requires careful attention to the posterior descending coronary artery during ablation. PMID- 19121800 TI - Left bundle branch-Purkinje system in patients with bundle branch reentrant tachycardia: lessons from catheter ablation and electroanatomic mapping. AB - BACKGROUND: The mechanism of bundle branch reentrant tachycardia has been described, and ablation of the right bundle branch (RBB) is a curative approach. OBJECTIVE: The purpose of this study was to evaluate the left bundle branch (LBB) Purkinje system during sinus rhythm and to test the feasibility of ablating the LBB in patients with bundle branch reentrant tachycardia. METHODS: Thirteen consecutive male patients (age 62 +/- 12 years) with sustained bundle branch reentrant tachycardia were included in the study. RESULTS: Surface ECG before ablation showed left bundle branch block (LBBB) in 10 patients and a narrow QRS in 3 patients. Bundle branch reentrant tachycardia with LBBB morphology was inducible in all cases. Ablation of the RBB resulted in right bundle branch block (RBBB) on surface ECG in 8 of 9 patients and total AV block with preserved retrograde conduction over the LBB in 1 of 9 patients. In 4 patients with LBBB during sinus rhythm, electroanatomic mapping showed (1) absent conduction through the anterior fascicle of the LBB and (2) anterograde slow conduction through the posterior fascicle of the LBB with (3) left ventricular activation by transseptal conduction due to conduction block between the distal Purkinje and the local ventricle in 2 of 4 patients. The LBB was successfully ablated in these 4 patients. During mean follow-up of 48 +/- 29 months, 3 patients died, but bundle branch reentrant tachycardia did not recur in any patient. CONCLUSION: In patients with LBBB and bundle branch reentrant tachycardia, anterograde slow conduction over the LBB is present. Ablation of the LBB is feasible and may be an alternative approach for bundle branch reentrant tachycardia. Patients with normal heart and bundle branch reentrant tachycardia have a good prognosis and may not require further intervention. PMID- 19121801 TI - Characterization of the phrenic nerve course within the epicardial substrate of patients with nonischemic cardiomyopathy and ventricular tachycardia. AB - BACKGROUND: Patients with nonischemic cardiomyopathy and ventricular tachycardia (VT) often have low-voltage areas in the lateral left ventricular (LV) epicardium that serve as the VT substrate. The course of the left phrenic nerve in this region may pose a challenge to successful and safe ablation. OBJECTIVE: The purpose of this study was to delineate the left phrenic nerve course in patients with nonischemic cardiomyopathy and suspected epicardial VT and to characterize its relationship with the VT substrate. METHODS: In 10 patients with nonischemic cardiomyopathy undergoing epicardial VT mapping and ablation, the course of the phrenic nerve was defined by pacing. The extent of epicardial LV low-voltage areas (<1.0 mV) was characterized by electroanatomic voltage mapping. RESULTS: Eight of 10 patients had low-voltage areas involving the lateral epicardial LV, and 7 of these 8 patients had sites of phrenic capture within these areas. Ablation was limited due to location of the phrenic nerve in two patients. In one of these patients, a balloon catheter was successfully used to mechanically protect the phrenic nerve during ablation. In the other five patients, adjacent ablation sites were targeted at which no phrenic capture with high-output pacing was demonstrated prior to ablation. In all patients undergoing ablation, the targeted VT became noninducible, and no patient demonstrated phrenic nerve injury. CONCLUSION: In most patients with nonischemic cardiomyopathy undergoing epicardial VT ablation, the phrenic nerve courses through a lateral LV low voltage area in proximity to potential sites for ablation. Strategies to identify and protect the phrenic nerve are important. PMID- 19121802 TI - Do abandoned leads pose risk to implantable cardioverter-defibrillator patients? AB - BACKGROUND: With the increased number of implantable cardioverter-defibrillator (ICD) recipients and the frequent need for device upgrading, lead malfunction is a concern, but the optimal approach to managing nonfunctioning leads is unknown. OBJECTIVE: The purpose of this study was to determine the rate and characteristics of complications related to abandoned ICD leads. METHODS: Patients with abandoned leads were identified by retrospective review of the Mayo Clinic ICD database from August 1993 to May 2002. We reviewed the medical records to assess long-term follow-up for venous thromboembolic complications, device sensing malfunction, appropriateness of delivered shocks, defibrillation threshold (DFT) values before and after lead abandonment, and subsequent surgical procedures related to devices or leads. RESULTS: We identified 78 ICD patients (81% males; mean age 63 +/- 14 years) with 101 abandoned leads (69 in the right ventricle, 31 in the right atrium or superior vena cava, 1 in the coronary sinus). During a mean follow-up of 3.1 +/- 2.0 years, neither sensing malfunction nor venous thromboembolic complications were detected. DFT values were high in 13 patients (17%), but there was no significant increase in mean DFT values before and after lead abandonment in 43 patients for whom both values were available (16.2 +/- 9.2 J before abandonment vs 14.1 +/- 5.5 J after; P = .24). Fourteen patients (18%) required further ICD-related surgery; none of these operations were attributed to abandoned leads. Five-year rates of appropriate and inappropriate shocks were 25.9% and 20.5%, respectively. CONCLUSION: Abandoning a nonfunctioning lead appears to be safe and does not pose a clinically significant additional risk of future complications. PMID- 19121804 TI - Reversal of neural and electrophysiologic remodeling in cardiac tissue. PMID- 19121803 TI - Effects of simvastatin on cardiac neural and electrophysiologic remodeling in rabbits with hypercholesterolemia. AB - BACKGROUND: Significant cardiac neural and electrophysiologic remodeling occurs with hypercholesterolemia (HC). Whether simvastatin can reverse HC-induced remodeling is unclear. OBJECTIVE: The purpose of this study was to determine the mechanisms underlying the antiarrhythmic effects of statins. METHODS: Rabbits (N = 38) were fed HC chow (HC), standard chow (Control), HC chow followed by standard chow (Withdrawal), or HC chow and simvastatin (Statin) for 8 weeks. The hearts then were Langendorff-perfused for electrophysiologic studies. Nerves were identified by immunostaining of growth-associated protein-43 (GAP43) and tyrosine hydroxylase (TH). Action potential duration (APD) restitution in normal hearts with (N = 5) and without (N = 5) simvastatin therapy also was studied. RESULTS: Serum cholesterol levels (mg/dL) were 1,855 +/- 533 in HC, 50 +/- 21 in Control, 570 +/- 115 in Withdrawal, and 873 +/- 112 in Statin groups (P <.001). Compared with HC (16,700 +/- 5,342; 12,200 +/- 3,878 microm(2)/mm(2)), the Statin group had significantly reduced GAP43-positive (10,289 +/- 3,393 microm(2)/mm(2), P = .03) and TH-positive (7,685 +/- 2,959 microm(2)/mm(2), P = .04) nerve density, respectively. APD was longer in HC rabbits than in controls (192 +/- 20 ms vs 174 +/- 17 ms; P <.03). Withdrawal and Statin groups had less APD prolongation than HC group. Statin group has less repolarization heterogeneity than HC group (P <.01). Statin therapy flattened the slope of APD restitution in normal hearts. Ventricular fibrillation was either induced or occurred spontaneously in 79% of hearts in HC, 20% in Control, and 66% in Withdrawal groups. However, there was no VF in hearts of Statin group (P <.001). CONCLUSION: Simvastatin significantly reduced vulnerability to ventricular fibrillation via the mechanism of reduction of HC-induced neural and electrophysiologic remodeling. PMID- 19121805 TI - Diacylglycerol kinase zeta inhibits G(alpha)q-induced atrial remodeling in transgenic mice. AB - BACKGROUND: Our previous study showed that diacylglycerol kinase zeta (DGKzeta), which degenerates diacylglycerol (DAG), inhibits ventricular structural remodeling and rescues activated G protein (alpha)q (G(alpha)q)-induced heart failure. However, whether DGKzeta inhibits atrial remodeling is still unknown. OBJECTIVE: This study aimed to elucidate the effects of DGKzeta on atrial remodeling. METHODS: A transgenic mouse (G(alpha)q-TG) with cardiac expression of activated G(alpha)q and a double transgenic mouse (G(alpha)q/DGKzeta-TG) with cardiac overexpression of DGKzeta and activated G(alpha)q were created. RESULTS: During electrocardiogram (ECG) recording for 10 min, atrial fibrillation was observed in 5 of 11 anesthetized G(alpha)q-TG mice but not in any wild-type (WT) and G(alpha)q/DGKzeta-TG mice (P <.05). All of the ECG parameters measured were prolonged in the G(alpha)q-TG compared with WT mice. Interestingly, in G(alpha)q/DGKzeta-TG mice, although the PR and RR intervals were still prolonged, the P interval, QRS complex, and QT interval were not different from those in WT mice. In Langendorff-perfused hearts, the incidence of atrial tachyarrhythmia induced by rapid atrial pacing was greater in G(alpha)q-TG hearts than in G(alpha)q/DGKzeta-TG hearts (P <.05). Action potential duration prolongation and impulse conduction slowing were observed in G(alpha)q-TG atria compared with G(alpha)q/DGKzeta-TG atria. Dilatation of the left atrium with thrombus formation was observed in 9 G(alpha)q-TG hearts but not in any G(alpha)q/DGKzeta-TG hearts. Moreover, the degree of extensive interstitial fibrosis in the left atrium was greater in G(alpha)q-TG hearts than that in G(alpha)q/DGKzeta-TG hearts (P <.05). CONCLUSION: These results show that DGKzeta inhibits G(alpha)q-induced atrial remodeling and suggest that DGKzeta is a novel therapeutic target for atrial fibrillation. PMID- 19121806 TI - G-protein signaling and arrhythmogenic atrial remodeling: relevance to novel therapeutic targets in atrial fibrillation. PMID- 19121807 TI - Reentry in Cardioversion: 'We can see it from here...'. PMID- 19121808 TI - Antiarrhythmic effect of IKr activation in a cellular model of LQT3. AB - BACKGROUND: Long QT syndrome type 3 (LQT3) is an inherited cardiac disorder caused by gain-of-function mutations in the cardiac voltage-gated sodium channel, Na(v)1.5. LQT3 is associated with the polymorphic ventricular tachycardia torsades de pointes (TdP), which can lead to syncope and sudden cardiac death. The sea anemone toxin ATX-II has been shown to inhibit the inactivation of Na(v)1.5, thereby closely mimicking the underlying cause of LQT3 in patients. OBJECTIVE: The hypothesis for this study was that activation of the I(Kr) current could counteract the proarrhythmic effects of ATX-II. METHODS: Two different activators of I(Kr), NS3623 and mallotoxin (MTX), were used in patch clamp studies of ventricular cardiac myocytes acutely isolated from guinea pig to test the effects of selective I(Kr) activation alone and in the presence of ATX-II. Action potentials were elicited at 1 Hz by current injection and the cells were kept at 32 degrees C to 35 degrees C. RESULTS: NS3623 significantly shortened action potential duration at 90% repolarization (APD(90)) compared with controls in a dose-dependent manner. Furthermore, it reduced triangulation, which is potentially antiarrhythmic. Application of ATX-II (10 nM) was proarrhythmic, causing a profound increase of APD(90) as well as early afterdepolarizations and increased beat-to-beat variability. Two independent I(Kr) activators attenuated the proarrhythmic effects of ATX-II. NS3623 did not affect the late sodium current (I(NaL)) in the presence of ATX-II. Thus, the antiarrhythmic effect of NS3623 is likely to be caused by selective I(Kr) activation. CONCLUSION: The present data show the antiarrhythmic potential of selective I(Kr) activation in a cellular model of the LQT3 syndrome. PMID- 19121810 TI - Successful ablation of epicardial arrhythmias with cryoenergy after failed attempts with radiofrequency energy. PMID- 19121809 TI - Restoring repolarization in LQT3. PMID- 19121811 TI - All LQT3 patients need an ICD: true or false? PMID- 19121812 TI - LQT3: who is at risk for sudden cardiac death? PMID- 19121813 TI - Molecular basis of catecholaminergic polymorphic ventricular tachycardia. AB - Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a malignant arrhythmia syndrome linked to mutations in the cardiac ryanodine receptor (RyR2) and calsequestrin (CASQ2). RyR2 and CASQ2 are parts of the multimolecular Ca(2+) release channel complex that is present on the sarcoplasmic reticulum (SR) to support myocyte Ca(2+) cycling and contractile activity. Whereas RyR2 operates as a Ca(2+) release channel, the SR Ca(2+) binding protein CASQ2 plays a dual role by serving as a SR Ca(2+) buffer and by regulating RyR2 function. Essential to stable Ca(2+) cycling, SR luminal Ca(2+)-dependent control of RyR2 activity by CASQ2 contributes to RyR2 deactivation and to the development of a temporary refractory state that occurs after each Ca(2+) release. Accumulating evidence suggests that the CPVT mutations act by reducing the extent and shortening the duration of Ca(2+) signaling refractoriness, thereby promoting untimely SR Ca(2+) release and arrhythmogenic delayed afterdepolarizations in cardiac myocytes. Similar mechanisms may apply to arrhythmias during various conditions, including heart failure and ischemic heart disease, associated with acquired defects in components of the Ca(2+) release channel complex. PMID- 19121814 TI - How to perform magnetic resonance imaging on patients with implantable cardiac arrhythmia devices. PMID- 19121815 TI - Roles of F-BAR/PCH proteins in the regulation of membrane dynamics and actin reorganization. AB - The Pombe Cdc15 Homology (PCH) proteins have emerged in many species as important coordinators of signaling pathways that regulate actomyosin assembly and membrane dynamics. The hallmark of the PCH proteins is the presence of a Fes/CIP4 homology Bin/Amphiphysin/Rvsp (F-BAR) domain; therefore they are commonly referred to as F BAR proteins. The prototype F-BAR protein, Cdc15p of Schizosaccharomyces pombe, has a role in the formation of the contractile actomyosin ring during cytokinesis. Vertebrate F-BAR proteins have an established role in binding phospholipids and they participate in membrane deformations, for instance, during the internalization of transmembrane receptors. This way the F-BAR proteins will function as linkers between the actin polymerization apparatus and the machinery regulating membrane dynamics. Interestingly, some members of the F-BAR proteins are implicated in inflammatory or neurodegenerative disorders and the observations can be expected to have clinical implications for the treatment of the diseases. PMID- 19121816 TI - New insights into the roles of dendritic cells in intestinal immunity and tolerance. AB - Dendritic cells (DCs) play a critical key role in the initiation of immune responses to pathogens. Paradoxically, they also prevent potentially damaging immune responses being directed against the multitude of harmless antigens, to which the body is exposed daily. These roles are particularly important in the intestine, where only a single layer of epithelial cells provides a barrier against billions of commensal microorganisms, pathogens, and food antigens, over a huge surface area. In the intestine, therefore, DCs are required to perform their dual roles very efficiently to protect the body from the dual threats of invading pathogens and unwanted inflammatory reactions. In this review, we first describe the biology of DCs and their interactions with other cells types, paying particular attention to intestinal DCs. We, then, examine the ways in which this biology may become misdirected, resulting in inflammatory bowel disease. Finally, we discuss how DCs potentiate immune responses against viral, bacterial, parasitic infections, and their importance in the pathogenesis of prion diseases. We, therefore, provide an overview of the complex cellular interactions that affect intestinal DCs and control the balance between immunity and tolerance. PMID- 19121817 TI - Transcriptional control of gene expression by actin and myosin. AB - Recent years have witnessed a new turn in the field of gene expression regulation. Actin and an ever-growing family of actin-associated proteins have been accepted as members of the nuclear crew, regulating eukaryotic gene transcription. In complex with heterogeneous nuclear ribonucleoproteins and certain myosin species, actin has been shown to be an important regulator in RNA polymerase II transcription. Furthermore, actin-based molecular motors are believed to facilitate RNA polymerase I transcription and possibly downstream events during rRNA biogenesis. Probably these findings represent the tip of the iceberg of a rapidly expanding area within the functional architecture of the cell nucleus. Further studies will contribute to clarify how actin mediates nuclear functions with a glance to cytoplasmic signalling. These discoveries have the potential to define novel regulatory networks required to control gene expression at multiple levels. PMID- 19121818 TI - Roles of eukaryotic Lsm proteins in the regulation of mRNA function. AB - The eukaryotic Lsm proteins belong to the large family of Sm-like proteins, which includes members from all organisms ranging from archaebacteria to humans. The Sm and Lsm proteins typically exist as hexameric or heptameric complexes in vivo and carry out RNA-related functions. Multiple complexes made up of different combinations of Sm and Lsm proteins are known in eukaryotes and these complexes are involved in a variety of functions such as mRNA decay in the cytoplasm, mRNA and pre-mRNA decay in the nucleus, pre-mRNA splicing, replication dependent histone mRNA 3'-end processing, etc. While most Lsm proteins function in the form of heteromeric complexes that include other Lsm proteins, some Lsm proteins are also known that do not behave in that manner. Abnormal expression of some Lsm proteins has also been implicated in human diseases. The various roles of eukaryotic Lsm complexes impacting mRNA function are discussed in this review. PMID- 19121819 TI - Spatiotemporal dynamics of the ER-derived peroxisomal endomembrane system. AB - Recent studies have provided evidence that peroxisomes constitute a multicompartmental endomembrane system. The system begins to form with the targeting of certain peroxisomal membrane proteins to the ER and their exit from the ER via preperoxisomal carriers. These carriers undergo a multistep maturation into metabolically active peroxisomes containing the entire complement of peroxisomal membrane and matrix proteins. At each step, the import of a subset of proteins and the uptake of certain membrane lipids result in the formation of a distinct, more mature compartment of the peroxisomal endomembrane system. Individual peroxisomal compartments proliferate by undergoing one or several rounds of division. Herein, we discuss various strategies that evolutionarily diverse organisms use to coordinate compartment formation, maturation, and division in the peroxisomal endomembrane system. We also critically evaluate the molecular and cellular mechanisms governing these processes, outline the most important unanswered questions, and suggest directions for future research. PMID- 19121820 TI - New insights into melanosome transport in vertebrate pigment cells. AB - Pigment cells of lower vertebrates provide an excellent model to study organelle transport as they specialize in the translocation of pigment granules in response to defined chemical cues. This review will focus on the well-studied melanophore/melanocyte systems in fish, amphibians, and mammals. We will describe the roles of melanin, melanophores, and melanocytes in animals, current views on how the three motor proteins dynein, kinesin, and myosin-V are involved in melanosome transport along microtubules and actin filaments, and how signal transduction pathways regulate the activities of the motors to achieve aggregation and dispersion of melanosomes. We will also describe how melanosomes are transferred to surrounding skin cells in amphibians and mammals. Comparative studies have revealed that the ability of physiological color change is lost during evolution while the importance of morphological color change, mainly via transfer of pigment to surrounding skin cells, increases. In humans, pigment mainly has a role in protection against ultraviolet radiation, but also perhaps in the immune system. PMID- 19121821 TI - Compartmentalizing the neuronal plasma membrane from axon initial segments to synapses. AB - Many membrane proteins localize to restricted domains in neurons, such as axons, dendrites, synapses, or axon initial segments. The exquisite subcellular compartmentalization of adhesion molecules, growth factor receptors, signaling receptors, voltage-gated and ligand-gated channels, and others underlies the complex functioning of neurons and ultimately vectorial propagation of signaling in neuronal circuits. This chapter discusses the cellular mechanisms for compartmentalizing the neuronal plasma membrane. Among the mechanisms contributing to protein segregation in the membrane are sorting and targeting in the Golgi/TGN, endocytosis, recycling, and degradation, and control of membrane protein diffusion. The molecular underpinnings of these cellular mechanisms are reviewed in the first part. The second part discusses the contribution of each cellular mechanism to targeting proteins to axons and dendrites, to synapses, to axon initial segments, and to Nodes of Ranvier. For most, if not all proteins and locations, all four mechanisms are in effect and additively contribute to the precise localization of membrane proteins in neurons. Since disruption of proper protein distribution results in defects in neuronal function and can lead to neurodegenerative diseases, a full understanding of the cellular mechanisms of plasma membrane compartmentalization is an important goal for the future. PMID- 19121822 TI - Influence of different artificial disc kinematics on spine biomechanics. AB - BACKGROUND: There are several different artificial discs for the lumbar spine in clinical use. Though clinically established, little is known about the biomechanical advantages of different disc kinematics. METHODS: A validated finite element model of the lumbosacral spine was used to compare the results of total disc arthroplasty at level L4/L5 performed by simulating the kinematics of three established artificial disc prostheses (Charite, ProDisc, Activ L). For flexion, extension, lateral bending, and axial torsion, the intervertebral rotations, the locations of the helical axes of rotation, the intradiscal pressures, and the facet joint forces were evaluated at the operated and adjacent levels. FINDINGS: After insertion of an artificial disc, intervertebral rotation is reduced for flexion and increased for extension, lateral bending, and axial torsion for all studied discs at implant level. The positions of the helical axes are altered especially for lateral bending and axial torsion. Increased facet joint contact forces are predicted for the Charite disc during extension-- influenced by the existence of anterior scar tissue--and for the ProDisc and the Activ L during lateral bending and axial torsion. The studied artificial discs have only a minor effect on the adjacent levels. INTERPRETATIONS: For some load cases, total disc arthroplasty leads to considerably altered kinematics and increased facet joint contact forces at implant level. The spinal kinematic alterations due to an artificial disc exceed by far the inter-implant differences, while facet joint contact force alterations are strongly implant and load case dependent. The importance of implant kinematics is often overestimated. PMID- 19121823 TI - Biomechanical comparison of traditional and minimally invasive intradural tumor exposures using finite element analysis. AB - BACKGROUND: Minimally invasive approaches to intradural pathology have evolved in part in an effort to reduce approach related destabilization of the spine. No biomechanical data exist however evaluating the effects of traditional and minimally invasive exposures. METHODS: A finite element model of the lumbar spine was generated, and a simulated open laminectomy and a modified hemilaminectomy at L4 were performed. Forces were applied to assess changes in flexion, extension, axial rotation, and lateral bending. FINDINGS: Open laminectomy produced much greater changes in extension, flexion, and axial rotation than the modified hemilaminectomy from the intact. Lateral bending was similarly unaffected for both exposures. INTERPRETATION: The results suggest that a minimally invasive hemilaminar exposure preserves the structural integrity of the lumbar spine and minimizes alterations to segmental motion postoperatively. PMID- 19121824 TI - Synthesis of bromo-conduritol-B and bromo-conduritol-C as glycosidase inhibitors. AB - For the synthesis of bromo-conduritol-B skeleton, bromo-1,4-benzoquinone was subjected to bromination followed by the reduction of the carbonyl groups with NaBH(4). Substitution of bromides bonded to sp(3)-hybridized carbon atoms with AgOAc gave the bromo-conduritol-B tetraacetate in high yield. For the construction of bromo-conduritol-C skeleton, 2,2-dimethyl-3a,7a-dihydro-1,3 benzodioxole was used as the starting material. Photooxygenation of the diene unit gave an unsaturated bicyclic endoperoxide. Bromine was incorporated into the molecule by the addition of bromine to the double bond. Opening of the peroxide linkage followed by HBr elimination and reduction of the carbonyl group provided the conduritol-C structure in good yield. Bromo-conduritol-B exhibited strong enzyme-specific inhibition against alpha-glycosidase. PMID- 19121825 TI - The effect of visual signals on spatial decision making. AB - We examined the effect of an irrelevant visual transient on the decision where to look for a hidden object. Participants also performed a conventional 'inhibition of return' localization task. In Experiments 1 and 2 the two tasks were blocked and in Experiments 3 and 4 they were randomly interleaved. In every experiment there was a bias to select the cued location in the spatial decision task. This facilitory effect was greatest when the cue occurred at a strategically unfavored location and even occurred for participants who reported strategically selecting a non-cued location, indicating that the facilitory effect is automatic and independent of other strategic biases. Inhibition of return was observed only when the tasks were blocked and the localization task preceded the decision task. The findings suggests that spatial decisions engage different attentional control settings than those engaged when detecting visual transients; and that this attentional mode affects the processing of visual transients such that they do not inhibit the subsequent speeded detection of onset targets. PMID- 19121827 TI - Surgical treatment for non-union of the mid-shaft clavicle using a reconstruction plate: scapular malposition is related to poor results. AB - A retrospective study was performed to evaluate the use of an AO reconstruction plate in open reduction and internal fixation for non-union of the mid-shaft clavicle, examining the relationship between the position of the scapula and final functional results and whether perioperative variables such as clavicular reconstruction ratio and period of non-union influence the position of the scapula. From January 1998 to January 2005, data on 21 people with symptomatic non-union of the mid-shaft clavicle were collected; 17 non-unions were atrophic and 4 were hypertrophic. Initially, treatment was conservative with a figure-of eight bandage in 19 cases, and cerclage wire fixation in 2 cases. The follow-up period was 65.7 (24-108) months. Outcome analyses included standard clinical follow-up, plain radiography, the Constant-Murley scoring and subjective assessment. All non-unions united well in 13.6 (11-27) weeks. All patients were satisfied with their surgical results. Greater age and longer period of non-union resulted in a larger amount of scapular malposition, which related to poor functional results. PMID- 19121828 TI - Injuries from leopard attacks in Kashmir. AB - Increased incidence of wild-animal attacks has made these a serious threat to public health in Kashmir. To establish patterns of injury and management, 35 case histories were reviewed. After resuscitation, of paramount importance were vigorous irrigation, removal of all foreign material, administration of antibiotics, and prophylaxis against rabies and tetanus as indicated. A multidisciplinary approach and aggressive management are necessary to prevent death and provide acceptable cosmetic and functional results. PMID- 19121826 TI - Sensitivity to syntax in visual cortex. AB - One of the most intriguing findings on language comprehension is that violations of syntactic predictions can affect event-related potentials as early as 120 ms, in the same time-window as early sensory processing. This effect, the so-called early left-anterior negativity (ELAN), has been argued to reflect word category access and initial syntactic structure building (Friederici, 2002). In two experiments, we used magnetoencephalography to investigate whether (a) rapid word category identification relies on overt category-marking closed-class morphemes and (b) whether violations of word category predictions affect modality-specific sensory responses. Participants read sentences containing violations of word category predictions. Unexpected items varied in whether or not their word category was marked by an overt function morpheme. In Experiment 1, the amplitude of the visual evoked M100 component was increased for unexpected items, but only when word category was overtly marked by a function morpheme. Dipole modeling localized the generator of this effect to the occipital cortex. Experiment 2 replicated the main results of Experiment 1 and eliminated two non-morphology related explanations of the M100 contrast we observed between targets containing overt category-marking and targets that lacked such morphology. Our results show that during reading, syntactically relevant cues in the input can affect activity in occipital regions at around 125 ms, a finding that may shed new light on the remarkable rapidity of language processing. PMID- 19121829 TI - Fertility assistance program following the Sichuan earthquake in China. AB - On May 12, 2008 an earthquake measuring 8.0 on the Richter scale hit Wenchuan in Sichuan province, a mountainous region in western China, and rendered many parents childless. Doctors from the Women's Fertility Program of the Second West China Hospital in Sichuan carried out a post earthquake fertility assistance program in the following 3 months. The program included 3 parts: sociomedical fertility health support; psychological counseling and intervention in reproductive health; and an assisted reproductive technology (ART) service. Couples whose children had died or were disabled in the earthquake were entitled to free fertility counseling and ART, if required. Some women are currently pregnant as a result of the program, which has been successful in helping some surviving couples achieve their reproductive desires. PMID- 19121830 TI - The active and passive ciliary motion in the embryo node: a computational fluid dynamics model. AB - The breaking of left-right symmetry in the mammalian embryo is believed to occur in a transient embryonic structure, the node, when cilia create a leftward flow of liquid. The two-cilia hypothesis proposes that the node contains two kinds of primary cilia: motile cilia that rotate autonomously to generate the leftward fluid flow and passive cilia that act as mechano-sensors, responding to flow. While studies support this hypothesis, the mechanism by which the sensory cilia respond to the fluid flow is still unclear. In this paper, we present a computational model of two cilia, one active and one passive. By employing computational fluid dynamics, deformable mesh computational techniques and fluid structure interaction analysis, and solving the three-dimensional unsteady transport equations, we study the flow pattern produced by the movement of the active cilium and the response of the passive cilium to this flow. Our results reveal that clockwise rotation of the active cilium can generate a counter clockwise elliptical rotation and overall lateral displacement for its neighboring passive one, of measurable magnitude and consistent pattern. This supports the plausibility of the two-cilia hypothesis and helps quantify the motion pattern for the passive cilium induced by this regional flow. PMID- 19121831 TI - Dynamic similarity during human running: about Froude and Strouhal dimensionless numbers. AB - Dynamic similarity is a widely used concept in the fluid mechanics field, and consists in placing two different-sized systems in equivalent experimental conditions. This enables removal of the effects of size and prediction of the behavior of a full size system from a scale model. The aim of this study was to test whether the Froude number (Nfr) or the Strouhal number (Str) could be used as a criterion for dynamic similarity during running. Fifteen male subjects ran barefoot on a runway in three experimental conditions (i) all subjects ran at the same speed V=3.5ms(-1); (ii) the speed was determined from Nfr; (iii) the stride frequency was determined from Str. Antero-posterior (Fy) and vertical (Fz) ground reaction force components were assessed. The similarity between the subjects was analysed from scale factor sets computed from anthropometric and kinetic data. The use of Str implied strong inter-subject similarity for temporal parameters (mean r=0.96, time to Fz peak, time to Fy braking peak, Fy zero fore-aft shear, time to Fy propulsive peak) while Nfr induced fewer and lower similarities (mean r=0.75, Fy zero fore-aft shear, time to Fy propulsive peak, Fy braking impulse) that only concerned antero-posterior parameters. This study brought experimental evidence that neither Nfr nor Str were sufficient for dynamic similarity during running, but that each of them made its own contribution. These findings suggested that the concomitant use of Nfr and Str should be assessed to induce inter-subject dynamic similarity during running. PMID- 19121832 TI - Free flow and capillary isoelectric focusing of bacteria from the tomatoes plant tissues. AB - The means of the preconcentration and preseparation of selected species or pathovars of bacteria directly from the plant tissue suspension by free flow isoelectric focusing are introduced here. After the focusing, the resulting fraction of microorganisms, native or dynamically modified by the non-ionogenic tenside on the basis of pyrenebutanoate, was separated by capillary isoelectric focusing and/or cultivated and positively identified by gas chromatographic analysis of fatty acid methyl esters. Simultaneously, capillary isoelectric focusing with UV and fluorometric detection was used for the rapid estimation of unknown isoelectric points of the examined plant pathogenic species of genus Clavibacter, Xanthomonas and Pseudomonas prior to the preconcentration and preseparation. The microorganisms were of different origin, native and/or dynamically modified by the non-ionogenic tenside. PMID- 19121833 TI - Integrated fluidic systems on a nanometer scale and the study on behavior of liquids in small confinement. AB - Nanofluidic systems and the studies on the behavior of liquids confined in nanometer-sized space are reviewed. Miniaturized chemical systems having nanometer-sized structures are fabricated by using advanced nanofabrication techniques. The size-confinement effect is expected to be applied in well controlled chemical and biochemical analysis. While electroosmosis and electrokinetic migration in small-sized channels have been investigated extensively, there have been few reports on pressure-driven flow systems having nanometer-sized structures, which are widely used in laboratory-scale and micrometer-sized systems. In this review, fundamental technologies that can be used in integrated chemical analysis systems having nanometer-sized structures are introduced. In addition to the technological investigations, important topics in the fundamental research on the properties of liquids confined in nanometer sized space are also presented. PMID- 19121834 TI - Impulsive-disinhibited personality and serotonin transporter gene polymorphisms: association study in an inmate's sample. AB - The association between different impulsive-disinhibited personality traits with 5-HTTLPR and 5-HTTVNTR genetic polymorphisms was examined in an imprisoned male sample. Higher scores of the impulsive-disinhibited personality traits tended to be associated with carrying one or two copies of the 5-HTTPLR S allele (S/S homozygous and S/L heterozygous), and carrying two copies of the 5-HTTVNTR 12 allele (12/12 homozygous). Genotype, allele, haplotype and extended genotype distribution between low and high impulsive-disinhibited groups confirmed this association. Allele S and genotypes S/S+S/L at the 5-HTTLPR locus and allele 12 and genotype 12/12 at the 5-HTTVNTR locus were overrepresented in the high scoring group. Accordingly, allele S and allele 12 conferred a trend for risk to be in the high scoring group with an odds ratio (OR) of 1.8 (p < 0.035) and 1.7 (p < 0.014), respectively. In addition, extended genotype distribution shows that those S allele carriers (S/S homozygote and S/L heterozygote) that were also 12/12 homozygote, were overrepresented in the high scoring group (OR = 3.2; p < 0.004). The main risk of being in the high scoring group was assigned to those carrying two copies of the S-12 haplotype (OR = 5.7; p < 0.0007). We discuss the possible relationship between the two genetic serotonin polymorphisms and the personality impulsive-disinhibited traits investigated. PMID- 19121835 TI - Lead-free KNLNT piezoelectric ceramics for high-frequency ultrasonic transducer application. AB - This paper presents the latest development of a lead-free piezoelectric ceramic and its application to transducers suitable for high-frequency ultrasonic imaging. A lead-free piezoelectric ceramic with formula of (K(0.5)Na(0.5))(0.97)Li(0.03)(Nb(0.9) Ta(0.1))O(3) (abbreviated as KNLNT 0.03/0.10) was fabricated and characterized. The material was found to have a clamped dielectric constant epsilon(33)(S)/epsilon(0)=890, piezoelectric coefficient d(33)=245 pC/N, electromechanical coupling factor k(t)=0.42 and Curie temperature T(c)>300 degrees C. High-frequency (40 MHz) ultrasound transducers were successfully fabricated with the lead-free material. A representative lead free transducer had a bandwidth of 45%, two-way insertion loss of -18 dB. This performance is comparable to reported performances of popular lead-based transducers. The comparison results suggest that the lead-free piezoelectric material may serve as an alternative to lead-based piezoelectric materials for high-frequency ultrasonic transducer applications. PMID- 19121836 TI - Declining levels of PCB, HCB and p,p'-DDE in adipose tissue from food producing bovines and swine in Sweden 1991-2004. AB - The official control programme for organochlorine (OC) contaminants in food producing animals in Sweden was used to study temporal and spatial trends of the polychlorinated biphenyl CB 153, hexachlorobenzene (HCB) and p,p'-DDE in adipose tissue from bovines and swine 1991-2004. Our results show that efforts to decrease OC contamination of animal feed and the environment have had a positive impact on the contamination of the animal production. OC concentrations declined significantly in almost all studied regions of Sweden. OC temporal trends were slower in bovines (6-8% per year) than in swine (10-12%). Power analyses showed that data from more than 10 years of sampling were needed for a detection of an annual OC level change of 5% in both species in the control programme, due to large within- and between-year variation in OC levels. CB 153 and p,p'-DDE levels were higher in southern than in northern Sweden. Levels decreased with age in milk cows, but not in young nulliparous cows (heifers) and bulls. Moreover, milk cows and bulls had significantly lower OC levels than heifers. Levels were not age-dependent among swine, but castrated male swine (barrows) had significantly lower OC levels than young female swine (gilts). Levels of the studied OCs are now in many cases below the LOQ of the analytical method used. Future time trend studies of these OCs thus depend on lowered LOQs in the control programme. PMID- 19121837 TI - Oxidative stress and histopathology damage related to the metabolism of dodecylbenzene sulfonate in Senegalese sole. AB - Surfactants such as linear alkylbenzene sulfonates (LAS) are widely utilised in the formulation of detergents in commercial products. After use, they pass through waste water treatment plants (WWTP) and are then discharged to aquatic ecosystems, causing risk to aquatic life. The exposure of marine animals to these compounds enhances the production of reactive oxygen species (ROS) with subsequent damage to macromolecules, and produces histological alterations. A flow-through experiment with Senegalese sole (Solea senegalensis) has been devised with the object of correlating the metabolism of LAS including sulfophenylcarboxylic acids (SPCs) by fish with their antioxidant defence system (generation of oxyradicals) and histopathological damage. The generation of intermediate degradation products (SPCs) by the organism, the histopathological responses, the antioxidant enzymes (catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione S-transferase (GST)), as well as other kinds of enzyme such as acid and alkaline phosphatases (AcP, ALP), were measured. SPCs from 5OC(6) to 11OC(12) were identified and quantified in fish and water; their concentrations differed depending on the sampling moment. In general, the responses found in the enzymes were slight: a decrease in the enzymatic activity in gills and activation in the digestive tract. The evidence of histopathological damage identified was also small; the organism's defensive mechanism against pollutants should enable it to recover easily. A direct relationship was established between biotransformation and the generation of SPCs and ROS. In conclusion, the correct functioning of the antioxidant defence system with absence of large variations, the short-term histopathological damage, and the evidence of SPCs indicate an adequate metabolism of 2-phenyl-C(12)-linear alkylbenzene sulfonates (2OC(12)LAS) by this specie and non-toxic effects at environmentally realistic levels. PMID- 19121839 TI - Toxicity of pulp and paper solid organic waste constituents to soil organisms. AB - This study examined the potential biological hazard of pulp and paper waste solids. The solids examined were chosen on the basis of the range of wood-related organic extractives and were either primary solids screened from the effluent stream before secondary treatment, or biosolids from aerated stabilisation lagoons. Acute effects were tested at the level of plants, invertebrates and soil microbes using an oat germination and growth test, earthworm survival and reproduction test, an enchytraeid worm survival and reproduction test, and standard measures of microbial respiration. This was further benchmarked against a marine bacteria toxicity test using extract of the waste solids. Resin acids and resin acid neutrals made up the greatest proportion of organic extractives measured in biosolids whereas resin acids and fatty acids were the main constituents detected in primary solids. Examination of the tissue of earthworms from the tests revealed no net bioconcentration of the organic extractives. The waste solids were not acutely toxic to any of the soil organisms as tested without any dilution. Conversely, extracts of the waste solids demonstrated toxicity in the marine bacteria. In some cases, the solid waste material enhanced the growth of plants, earthworm reproduction and microbial respiration. The only adverse affect was that reproduction of enchytraeids was reduced by some of the waste solid treatments. However these effects did not appear to be associated with concentrations of resin acid neutrals and resin acids in these materials. Overall pulp and paper wastes were relatively benign in terms of toxicity to the soil organisms tested. PMID- 19121838 TI - Contamination profiles and mass loadings of macrolide antibiotics and illicit drugs from a small urban wastewater treatment plant. AB - Information is limited regarding sources, distribution, environmental behavior, and fate of prescribed and illicit drugs. Wastewater treatment plant (WWTP) effluents can be one of the sources of pharmaceutical and personal care products (PPCP) into streams, rivers and lakes. The objective of this study was to determine the contamination profiles and mass loadings of urobilin (a chemical marker of human waste), macrolide antibiotics (azithromycin, clarithromycin, roxithromycin), and two drugs of abuse (methamphetamine and ecstasy), from a small (<19 mega liters day(-1), equivalent to <5 million gallons per day) wastewater treatment plant in southwestern Kentucky. The concentrations of azithromycin, clarithromycin, methamphetamine and ecstasy in wastewater samples varied widely, ranging from non-detects to 300 ng L(-1). Among the macrolide antibiotics analyzed, azithromycin was consistently detected in influent and effluent samples. In general, influent samples contained relatively higher concentrations of the analytes than the effluents. Based on the daily flow rates and an average concentration of 17.5 ng L(-1) in the effluent, the estimated discharge of azithromycin was 200 mg day(-1) (range 63-400 mg day(-1)). Removal efficiency of the detected analytes from this WWTP were in the following order: urobilin>methamphetamine>azithromycin with percentages of removal of 99.9%, 54.5% and 47%, respectively, indicating that the azithromycin and methamphetamine are relatively more recalcitrant than others and have potential for entering receiving waters. PMID- 19121840 TI - Duodenal epithelioid gastrointestinal stromal tumor with prominent granular cell features. AB - Gastrointestinal stromal tumors are a group of mesenchymal tumors arising from the wall of the gastrointestinal tract that are characterized by activating mutations in KIT or PDGFRA. Their proper recognition is important clinically because of their potential responsiveness to targeted therapies. We report a case of duodenal gastrointestinal stromal tumor with a highly unusual epithelioid morphology that had an appearance reminiscent of a steroid producing neoplasm, such as an adrenal cortical neoplasm or, alternatively, a renal cell carcinoma variant. The recognition of the current tumor as a duodenal gastrointestinal stromal tumor was prompted by its apparent location in the duodenal wall. Ancillary immunohistochemical and molecular sequence analyses were necessary to confirm the diagnosis as a gastrointestinal stromal tumor. The current case illustrates the importance of considering gastrointestinal stromal tumor in the differential diagnosis of any epithelioid tumors in the gastrointestinal tract or the abdominal-pelvic cavity. PMID- 19121841 TI - Characterization of lymphangiogenesis in various stages of idiopathic diffuse alveolar damage. AB - In pulmonary fibrosis, an abnormal healing process, is believed to be involved in the damage to lung tissue. This process has not been correlated with lymphangiogenesis, which has garnered current interest in relation to wound healing. The aim of the present study was to clarify the characteristics of lymphangiogenesis in pulmonary fibrosis associated with idiopathic diffuse alveolar damage. Formalin-fixed and paraffin-embedded lung tissues from 13 autopsy cases with idiopathic diffuse alveolar damage were used. Antibodies specific to CD34 and D2-40 were used to detect blood vessels and lymphatics, respectively, and immunohistochemical examinations and morphometry analyses were performed. The standardized density of capillaries was increased significantly in the exudative stage of diffuse alveolar damage, whereas that of the lymphatics remained unchanged. In the proliferative stage, new lymphatics emerged, primarily in the intra-alveolar fibrotic lesions where capillaries were absent. In the fibrotic stage, in which the lung was shrunken, as revealed by the elevated density of pulmonary arteries, the standardized density of capillaries was reduced significantly. The standardized area density of the interstitium was elevated in the proliferative stage and subsequently reduced in the fibrotic stage. Three-dimensional reconstruction of images revealed that some new lymphatics lacked connection to existing lymphatics. During the progression of diffuse alveolar damage, lymphangiogenesis occurs independent of capillary angiogenesis. PMID- 19121842 TI - Quantitation of T lymphocytes in posttransplant transbronchial biopsies. AB - The diagnostic role of immunohistochemical staining for T lymphocytes in grading acute airway rejection has not been fully explored. We examined 136 transbronchial biopsies from 52 lung transplant patients and 9 nontransplant controls. Transplant rejection was based on histologic assessment of perivascular (A) and bronchiolar (B) infiltrates. The clinical indication for the 136 allograft biopsies was routine surveillance (n = 72), decreased pulmonary function, rule out rejection (n = 36), suspect infection (n = 16), rule out obliterative bronchiolitis (n = 6), and persistent postoperative graft failure (n = 6). T lymphocytes were counted in bronchial mucosa per 100 bronchial epithelial cells, and in alveolar walls per square millimeters, after immunohistochemical staining with anti-CD3, CD4, and CD8. In controls, the mean alveolar wall CD3 cell count was 45 per square millimeter (95% confidence intervals, 30-52 per square millimeter) and the mean CD8 count was 15 per square millimeter (2-20 per square millimeter). In surveillance and negative patient biopsies, alveolar wall CD8 counts were significantly greater than controls (P = .03 and .02, respectively). Mean alveolar wall CD3 counts were significantly higher in type A rejection (88.7 +/- 12.9) than controls and negative biopsies (42 +/- 5.3, P < .001), but there was no difference compared to infections (119.7 +/- 22, P > .5). Mucosal CD3 cell counts were significantly higher in type B rejection (16.1 +/- 2.5) than controls and negative biopsies (1.5 +/- 0.4, P < .001), and also higher than infections (3.9 +/- 1.1, P < .001). In 7% of biopsies, T-cell staining identified perivascular circumferential infiltrates that were difficult to identify on routine stains, and in an additional 9% minor changes in grading were made after reviewing T-cell markers. Immunohistochemical staining may help in identifying perivascular infiltrates and demonstrates increased intraepithelial T cells even in low-grade type B rejection. Type B rejection as assessed quantitatively is more specific than type A rejection in comparison to infection. PMID- 19121843 TI - Systemic infection of avian influenza A virus H5N1 subtype in humans. AB - The viral dissemination in a patient with avian influenza A subtype H5N1 infection was retrospectively studied by the immunohistochemical localization of viral nucleoprotein antigen. The pathology was marked by diffuse alveolar damage, lymphoid depletion, and reactive hemophagocytic syndrome. Besides the lung and the upper respiratory tract, viral antigen was detected in the small and large intestinal epithelial cells, hematopoietic cells in the bone marrow, glial cells and neurons of the brain, and lymphocytes. The results confirmed that H5N1 virus disseminated to multiple organs beyond the respiratory system. However, specific pathological changes were noted in the respiratory system only, and productive viral replication confirmed by culture was noted only in the lung. More postmortem studies are needed to elucidate the pathogenesis of this highly fatal zoonotic disease. PMID- 19121844 TI - Forkhead box M1 expression in pulmonary squamous cell carcinoma: correlation with clinicopathologic features and its prognostic significance. AB - Forkhead box M1 (FoxM1) transcription factor has been shown to play important roles in regulating the expression of genes that are involved in cell proliferation, differentiation, and transformation by promoting both G(1)/S and G(2)/M transition. Although it has been reported that the FoxM1 signaling network is frequently deregulated with an up-regulated FoxM1 expression in human malignancies, the role of FoxM1 in lung cancer remains to be determined. We performed immunohistochemical detection of FoxM1 protein in 69 tissue samples from patients with primary pulmonary squamous cell carcinoma using a tissue microarray, and Western blotting was done to confirm the immunohistochemical observations. FoxM1 immunoreactivity was observed in 26 (37.7%) of the 69 squamous cell carcinoma cases. Analysis of the FoxM1 expression in 12 squamous cell carcinoma tissues and 2 normal lung tissues by Western blotting confirmed the immunohistochemical results. A FoxM1 expression was more frequently detected in the moderately or poorly differentiated squamous cell carcinomas than in the well-differentiated squamous cell carcinomas (P = .008). The tumors with a positive FoxM1 expression more frequently showed lymph node metastasis (P = .027) and an advanced American Joint Committee on Cancer stage (P = .049). The Kaplan Meier survival curves demonstrated that patients with a positive FoxM1 expression had a significantly shorter survival time than those patients with a negative FoxM1 expression (P = .003). The multivariate analysis revealed that the FoxM1 expression was an independent poor prognostic factor (P = .018). A subset of pulmonary squamous cell carcinoma with a FoxM1 expression was associated with progressive pathologic features and an aggressive clinical course. PMID- 19121845 TI - Intraprostatic adipose tissue: a study of 427 whole mount radical prostatectomy specimens. AB - Prostatic adenocarcinoma is the most frequently diagnosed cancer in American men. Tumor Gleason grade and stage provide extremely valuable prognostic information and play an important role in therapeutic decision making and patient counseling. A biopsy or radical prostatectomy specimen revealing carcinoma extending into extraprostatic tissue permits a T3 classification. This is most easily recognized, particularly in a needle biopsy, when tumor is seen to invade the adipose tissue. The existence of intraprostatic adipose tissue is somewhat controversial. To investigate this, formalin-fixed paraffin-embedded whole-mount radical prostatectomy specimens from 427 patients with adenocarcinoma were evaluated for intraprostatic adipose tissue. It was defined as any collection of adipocytes amid or internal to the most peripheral glands. The amount, anatomic location, and relationship to normal structures were also recorded. Intraprostatic adipose tissue was identified in 17 (3.98%) of cases. It consisted of small microscopic foci composed of 5 to 20 adipocytes. In 13 cases, the fat was intimately associated with benign glands. In another 2 cases, it was associated with small nerves, and in 2 cases was random with no specific localization. Intraprostatic adipose tissue was located in the peripheral zone in 15 cases and in the central zone in 2. Intraprostatic adipose tissue, although uncommon, does exist. Therefore, caution must be exercised in diagnosing extraprostatic extension based only upon identification of fat invasion, especially in a needle biopsy. The small size of foci of adipose tissue and its admixture with benign glands are useful morphologic clues in distinguishing it from extraprostatic fat. PMID- 19121846 TI - Reduced beta-catenin and peroxisome proliferator-activated receptor-gamma expression levels are associated with colorectal cancer metastatic progression: correlation with tumor-associated macrophages, cyclooxygenase 2, and patient outcome. AB - Recent studies have reported cross talk between beta-catenin, peroxisome proliferator-activated receptor-gamma, and cyclooxygenase 2 signaling pathways. We examined whether molecular changes of these pathways could be related to colorectal cancer metastatic progression. Seventy-two sporadic colorectal cancers and the distant nonneoplastic mucosa were analyzed for beta-catenin, peroxisome proliferator-activated receptor-gamma, cyclooxygenase 2, and nuclear factor kappaB levels by immunohistochemistry and Western blot. The expression profiles were correlated with patient outcome and 5-year survival. Nuclear beta-catenin staining was detected in only 18.1% of tumors and correlated with poor survival as compared with cases showing cytosolic/membrane accumulation (59.7%, P < .05). This latter group and tumor samples showing cytosolic/nuclear peroxisome proliferator-activated receptor-gamma expression (70.8%) were significantly associated with a favorable prognosis (P < .001). Remarkably, reduction or loss of beta-catenin (22.2%) and peroxisome proliferator-activated receptor-gamma (29.2%) expression was strongly correlated with marked infiltration of tumor associated macrophages (P < .01), presence of liver metastases, and very short survival (P = .0001). Moreover, beta-catenin and peroxisome proliferator activated receptor-gamma levels were inversely correlated with cyclooxygenase 2 (P < .01) and nuclear factor kappaB expression (P < .05). Our results suggest that reduced expression of beta-catenin and peroxisome proliferator-activated receptor-gamma could play a key role in aggressive colorectal cancer behavior. This finding may provide a relevant prognostic tool and contribute to early identification of patients at high risk of mortality. PMID- 19121847 TI - Diagnostic and prognostic role of the insulin growth factor pathway members insulin-like growth factor-II and insulin-like growth factor binding protein-3 in serous effusions. AB - We recently reported on higher expression of the insulin-like growth factor pathway genes IGF-II and IGFBP3 in serous ovarian/peritoneal carcinoma compared to malignant peritoneal mesothelioma. The present study analyzed the diagnostic and clinical role of these proteins in serous effusions. Effusions (n = 327), including 294 carcinomas (205 ovarian, 48 breast, 17 cervical/endometrial, 12 lung, 12 gastrointestinal/genitourinary) and 33 malignant mesotheliomas, were immunostained for insulin-like growth factor-II and insulin-like growth factor binding protein-3. Surgical ovarian carcinoma (n = 124) and peritoneal mesothelioma (n = 18) specimens were additionally studied. Insulin-like growth factor binding protein-3 levels were measured in 148 effusion supernatants (114 ovarian carcinomas, 18 breast carcinomas, 16 mesotheliomas) using enzyme-linked immunosorbent assay. Insulin-like growth factor binding protein-3 promoter methylation was analyzed in 11 ovarian carcinoma effusions. Insulin-like growth factor binding protein-3 (P = .002) and insulin-like growth factor-II (P < .001) expression by immunohistochemistry was significantly higher in carcinomas compared to mesotheliomas, with diagnostic sensitivity of 77% and 70% and specificity of 55% and 70%, respectively. In surgical specimens, insulin-like growth factor binding protein-3 expression was higher in ovarian carcinomas compared to peritoneal mesotheliomas (P = .007), whereas insulin-like growth factor-II expression was comparable (P = .505). Insulin-like growth factor binding protein-3 levels by enzyme-linked immunosorbent assay were comparable in the 3 analyzed cancer types. Insulin-like growth factor binding protein-3 promoter methylation was found in 6 of 11 effusions. High insulin-like growth factor binding protein-3 expression in prechemotherapy and high insulin-like growth factor-II expression in postchemotherapy ovarian carcinoma effusions correlated with poor overall survival (P = .031 and P = .024, respectively). Insulin-like growth factor-II expression in postchemotherapy effusions was an independent prognostic factor in Cox multivariate analysis (P = .04). In conclusion, insulin-like growth factor-II and insulin-like growth factor binding protein-3 are more frequently expressed in metastatic carcinomas compared to mesothelioma in effusions but are less specific than currently used markers. Insulin-like growth factor-II and insulin-like growth factor binding protein-3 may be novel prognostic markers in metastatic ovarian carcinoma. PMID- 19121848 TI - Clinicopathologic spectrum of massive and submassive hepatic necrosis in infants and children. AB - Clinicopathologic features of 45 patients with fulminant hepatic failure due to massive or submassive hepatic necrosis were studied. Both percutaneous biopsies and liver explants were available in 23 patients, whole livers only in 11 cases, and biopsies only in 11 cases. An etiologic diagnosis was established in 16 cases (36%). A further 3 cases (7%) were associated with aplastic anemia. Established etiologies included drug reactions (n = 7); autoimmune hepatitis, type 2 (n = 3); halothane hepatitis (n = 1); ischemia/hypotension (n = 1); mushroom poisoning (n = 1); mitochondrial disorder (n = 1); hemophagocytic lymphohistiocytosis (n = 1); and adenoviral hepatitis (n = 1). The extent of necrosis on liver biopsy correlated poorly with that in liver explants (mean difference, 32% +/- 23.8%). Almost all cases could be classified into one of 2 broad patterns of necrosis, namely, (1) zonal coagulative necrosis or (2) panlobular (nonzonal) necrosis. These patterns differed significantly with respect to several clinical parameters including sex ratio, peripheral blood white cell count, serum aspartate transaminase and alanine transaminase, conjugated bilirubin, and alkaline phosphatase levels. Livers with panlobular necrosis showed a spectrum of histopathologic findings that included central venulitis (76%), lymphocytic infiltration of large duct/gallbladder epithelium (54%), and syncytial giant cell transformation (18%). These features were not seen in livers with zonal coagulative necrosis which frequently showed prominent steatosis (91%). Both patterns of necrosis frequently showed ductular proliferation (100%) and cholangiolitis (80%). The diagnostic yield of ancillary studies (histochemistry, immunohistochemistry, and electron microscopy) was very low (<1%). The small proportion of cases with etiologic diagnoses precluded correlation of clinical and histopathological parameters with specific etiologies. In summary, this study describes the spectrum of changes seen in massive and submassive necrosis in children and identifies clinical features that might differentiate between 2 broad patterns of necrosis. PMID- 19121849 TI - Phosphorylated hepatocyte growth factor receptor/c-Met is associated with tumor growth and prognosis in patients with bladder cancer: correlation with matrix metalloproteinase-2 and -7 and E-cadherin. AB - Hepatocyte growth factor receptor/c-Met is associated with malignant aggressiveness and survival in various cancers including bladder cancer. Although phosphorylation of hepatocyte growth factor receptor/c-Met is essential for its function, the pathologic significance of phosphorylated hepatocyte growth factor receptor/c-Met in bladder cancer remains elusive. We investigated the clinical significance of its expression, and its correlation with cancer cell progression related molecules. The expression levels of 2 tyrosine residues of hepatocyte growth factor receptor/c-Met (pY1234/1235 and pY1349) were examined immunohistochemically in 133 specimens with nonmetastatic bladder cancer. We also investigated their correlation with matrix metalloproteinase-1, -2, -7, and -14; urokinase-type plasminogen activator; E-cadherin; CD44 standard, variant 3, and variant 6; and vascular endothelial growth factor. Expression of phosphorylated hepatocyte growth factor receptor/c-Met was detected in cancer cells, but was rare in normal urothelial cells. Although hepatocyte growth factor receptor/c Met, pY1234/1235 hepatocyte growth factor receptor/c-Met, and pY1349 hepatocyte growth factor receptor/c-Met were associated with pT stage, multivariate analysis identified pY1349 hepatocyte growth factor receptor/c-met expression only as a significant factor for high pT stage. Expression of pY1349 hepatocyte growth factor receptor/c-Met was a marker of metastasis and (P = .001) and cause specific survival (P = .003). Expressions of matrix metalloproteinase-2, matrix metalloproteinase-7, and E-cadherin correlated with pY1349 hepatocyte growth factor receptor/c-Met expression. Our results demonstrated that pY1349 hepatocyte growth factor receptor/c-Met plays an important role in tumor development, and its expression is a significant predictor of metastasis and survival of patients with bladder cancer. The results suggest that these activities are mediated, at least in part, by matrix metalloproteinase-2, matrix metalloproteinase-7, and E cadherin. PMID- 19121850 TI - Deconvolution of trace element (As, Cr, Mo, Th, U) sources and pathways to surface waters of a gold mining-influenced watershed. AB - The Upper Isle River (SW France) drains the second most productive gold-mining district of France. A high resolution survey during one hydrological year of As, Cl(-), Cr, Fe, Mn, Mo, SO(4)(2-), Th and U dissolved concentrations in surface water aimed to better understand pathways of trace element export to the river system downstream from the mining district. Dissolved concentrations of As (up to 35000 ng/L) and Mo (up to 292 ng/L) were about 3-fold higher than the regional dissolved background and showed a negative logarithmic relation with discharge. Dissolved concentrations of Cr (up to 483 ng/L), Th (up to 48 ng/L) and U (up to 184 ng/L) increased with discharge. Geochemical relationships between molar ratios in surface water, geochemical background as well as rain- and groundwater data were combined. The contrasting behavior of distinct element groups was explained by a scenario involving three seasonal components: (i) The high flow component is poorly concentrated in As and Mo but highly concentrated in Cr, Th, U. This has been attributed to diffuse sources such as water-soil interactions, atmospheric inputs, bedrock and bed sediment weathering. Although this component probably also includes a contribution by weathering of sulfide veins, this signal is masked by dilution. (ii) One low flow component presents high SO(4)(2-), Fe, As and Mo and moderate Cr, Th and U concentrations. This component has been attributed to point sources such as mine gallery effluents, mining waste weathering and groundwater inputs from natural and/or mining-induced sulfide oxidation in the ore deposit. (iii) A second low flow component showing high As plus Mo concentrations associated with very low SO(4)(2-), Fe, Cr, Th and U concentrations, probably reflects trace element scavenging by ferric oxyhydroxide formation in the adjacent aquifer. This is supported by the decrease of Fe, Cr, Th and U in surface waters. Flux estimates suggest contrasting element-specific impacts on annual dissolved fluxes. Runoff may account for the major part of annual dissolved As, Mo, Th and U fluxes in the Upper Isle River. Inputs related to sulfide oxidation respectively contributed approximately 30% and approximately 24% to annual As and Mo fluxes. The formation of ferric oxyhydroxides strongly retained Cr, Th and U during the low flow, limiting their dissolved concentrations in surface waters. If this process may eventually decrease As mobility, its impact on dissolved As concentrations in surface water may be limited or/and counterbalanced by As release during sulfide oxidation. PMID- 19121851 TI - Nervous control of the gills. AB - The fish gill is a highly complex organ that performs a wide variety of physiological processes and receives extensive nervous innervation from both afferent (sensory) and efferent (motor) fibres. Innervation from the latter source includes autonomic nerve fibres of spinal (sympathetic) and cranial (parasympathetic) origin whose primary role is to induce vasomotor changes within the respiratory or nonrespiratory pathways of the gill vasculature. Autonomic control of the gill occurs by nerve fibres identified as adrenergic, cholinergic, and more recent evidence indicates that nonadrenergic-noncholinergic (NANC) nerve fibres, such as those that express amines, peptides, or nitric oxide, may also play an important role. The distribution and physiological function of NANC nerve fibres, however, is less clear. This review primarily discusses histochemical studies that have characterized the nervous innervation and autonomic control of the gill vasculature. In addition, supporting evidence from recent studies for the efferent control, or modulation, of other homeostatic processes in the gill is examined. PMID- 19121852 TI - Cardio-respiratory control during early development in the model animal zebrafish. AB - Independent of species, the cardiovascular system is the first functioning component of developing vertebrate embryos. One of the main hypotheses is the assumption that larval and juvenile stages of fish and amphibians are not just smaller versions of an adult phenotype. In this review, the cardiovascular and respiratory responses to environmental, genetic and epigenetic perturbations are discussed in detail to understand the relationships between cardiac and respiratory performance, haematopoiesis for embryonic or larval stages with special focus on the popular model animal, the zebrafish. Zebrafish are tiny animals which have many advantages as a model organism in analysis of the cardio respiratory system. It obtains sufficient amounts of oxygen via bulk diffusion, in contrast to convection-dependent mammals. It is possible to study genetic mutants even with extreme defective phenotypes of the cardio-respiratory system in order to understand its developmental and physiological mechanisms. It has become apparent that the cardio-respiratory system and its control starts functioning very early during development, long before oxygen uptake becomes diffusion limited in zebrafish. Finally, recent improvements in imaging techniques for the use of fish models relevant for developmental physiology and biomedical research are discussed. PMID- 19121853 TI - Imaging anisotropy using differential polarization laser scanning confocal microscopy. AB - We have constructed differential polarization (DP) attachments to a laser scanning microscope (LSM) for imaging the main DP quantities of anisotropic microscopic objects. The DP-LSM operates with high-frequency modulation and subsequent demodulation and displays the main DP quantities pixel by pixel. These, for linearly polarized light, include: (i) linear birefringence (LB), which is exhibited by structurally and/or optically anisotropic material; (ii) linear dichroism (LD), which carries information on the anisotropic distribution of the molecules, i.e. of their absorbance transition dipole vectors, in the sample; (iii) fluorescence-detected LD (FDLD), which carries the same information for fluorescent dyes upon excitations with two orthogonally polarized light beams; (iv) anisotropy of the fluorescence emission (r), excited with non polarized light, which is determined by the distribution of the emission transition dipole vectors in the sample and is analogous with LD and (v) the degree of polarization of the fluorescence emission (P), excited with polarized light, which depends on the depolarization of the emission e.g. due to the rotation of molecules during their excitation lifetimes. In fluorescence regimes, the DP images can be recorded in the confocal regime of the microscope, which thus warrants good spatial resolution and the possibility of mapping the anisotropy in three dimensions. In this paper, we outline the design and technical realization of our DP-LSM and give a few examples on DP imaging of different biological samples. PMID- 19121856 TI - Gene variants of IL13, IL4, and IL4RA are predictors of beta-lactam allergy. PMID- 19121858 TI - Contribution of histamine metabolism to tachyphylaxis during the buildup phase of rush immunotherapy. PMID- 19121859 TI - Respiratory syncytial virus: back to basics. PMID- 19121857 TI - Peanuts can contribute to anaphylactic shock by activating complement. AB - BACKGROUND: Peanut allergy is the most common food-related cause of lethal anaphylaxis and, unlike other food allergies, typically persists into adulthood. Resistance to digestion and dendritic cell activation by the major peanut allergen Ara h 1 are reported to contribute to its allergenicity. OBJECTIVE: We sought to evaluate whether peanut molecules might also promote anaphylaxis through an innate immune mechanism. METHODS: Naive mice were treated with a beta adrenergic receptor antagonist and long-acting IL-4 to increase sensitivity to vasoactive mediators and injected with peanut extract (PE). Shock was detected and quantified by means of rectal thermometry. Gene-deficient mice and specific antagonists were used to determine the roles of specific cell types, complement, Fc receptors, and vasoactive mediators in shock pathogenesis. RESULTS: PE induces dose-dependent shock. PE activates complement in vivo in mice and in vitro in mice and human subjects. C3a and, to a lesser extent, stimulatory immunoglobulin receptors contribute to PE-induced shock. PE-induced shock depends more on macrophages and basophils than on mast cells. Platelet-activating factor and, to a lesser extent, histamine contribute to PE-induced shock. PE induces shock in the absence of the adaptive immune system. LPS contamination is not responsible for PE-induced shock. PE and IgE-mediated mast cell degranulation synergistically induce shock. Tree nuts have similar effects to PE, and skim milk and egg white do not. CONCLUSION: Peanuts can contribute to shock by causing production of C3a, which stimulates macrophages, basophils, and mast cells to produce platelet activating factor and histamine. PMID- 19121861 TI - Airway wall structural remodeling from endobronchial biopsies: what does it really mean? PMID- 19121860 TI - Phenotypic predictors of long-term response to inhaled corticosteroid and leukotriene modifier therapies in pediatric asthma. AB - BACKGROUND: In children with mild-to-moderate persistent asthma, identification of phenotypic predictors to guide selection of a controller regimen is essential. OBJECTIVE: We sought to identify phenotypic characteristics having predictive value for the difference in treatment responses between twice-daily fluticasone and once-daily montelukast. METHODS: Data from the Pediatric Asthma Controller Trial were assessed with multivariate analysis. Outcomes included the change in asthma control days (ACDs), FEV(1), peak expiratory flow, and time to first asthma exacerbation measured over a 1-year treatment period. RESULTS: The mean age was 9.6 +/- 2.1 years, 60% were male, 50% had a parental history of asthma, and 78% had positive aeroallergen skin prick test responses. The mean percent predicted prebronchodilator FEV(1) was 97.8% +/- 12.9%, the median PC(20) value was 0.93 mg/mL, and the median exhaled nitric oxide (eNO) level was 25.2 ppb. A history of parental asthma best predicted the expected treatment benefit with fluticasone compared with montelukast in terms of gain in ACDs (adjusted P = .02) and time to first exacerbation (adjusted P = .05). Increased baseline eNO levels predicted the differential treatment response for fluticasone regarding the gain in ACDs (adjusted P = .01). Prior inhaled corticosteroid (ICS) use (adjusted P = .01) and low PC(20) values (adjusted P = .03) each predicted the expected treatment benefit with fluticasone over montelukast regarding time to first exacerbation. No phenotypic characteristics predicted treatment benefits for montelukast over fluticasone for either outcome. CONCLUSIONS: Physicians treating children with a parental history of asthma, increased eNO levels, low PC(20) values, or a history of ICS use can expect the best long-term outcomes with ICS therapy compared with treatment with leukotriene receptor antagonists. PMID- 19121862 TI - Impaired translation of CCAAT/enhancer binding protein alpha mRNA in bronchial smooth muscle cells of asthmatic patients. AB - BACKGROUND: Bronchial smooth muscle (BSM) cells of asthmatic patients have an impaired expression of CCAAT/enhancer binding protein (C/EBP) alpha, which is associated with increased proliferation. OBJECTIVE: We sought to assess the translational regulation of CEBPA mRNA in cultured BSM cells of healthy control subjects (n = 11) and asthmatic patients (n = 12). METHODS: Translation efficiency was studied by using a translation control reporter system driven by the control elements present in the CEBPA mRNA. Translation efficiency was determined by the ratio of 2 artificial hemagglutinin (HA.11) proteins: p23 and p12. We also analyzed levels of proteins that control translation of CEBPA mRNA, namely heterogeneous nuclear ribonucleoprotein E2, calreticulin, eukaryotic translation initiation factor (eIF4E), and 4E binding protein. RESULTS: Compared with healthy control subjects, BSM cells of asthmatic patients proliferate faster (2.1-fold) and are primed for IL-6 secretion. Real-time RT-PCR showed that BSM cells of asthmatic patients express normal levels of CEBPA mRNA, whereas they express lower levels of C/EBPalpha (p42). Transient transfections with the translation control reporter system construct showed a disturbed p12/p23 ratio in BSM cells of asthmatic patients relative to healthy control subjects, which coincided with lower levels of eIF4E. CONCLUSION: BSM cells of asthmatic patients have normal levels of CEBPA mRNA but inadequately reinitiate the translation into C/EBPalpha. Impaired translation control upstream of eIF4E might underlie the observed increased proliferation and priming of BSM cells of asthmatic patients. PMID- 19121863 TI - Oral contraception, body mass index, and asthma: a cross-sectional Nordic-Baltic population survey. AB - BACKGROUND: Emerging evidence suggests that sex steroid hormones may influence airways obstruction, and that metabolic status may modify potential effects. OBJECTIVE: This study investigated the association between use of oral contraceptive pills (OCPs) and asthma in a Nordic-Baltic population-based study, while taking into account possible interplay with body mass index (BMI). METHODS: Postal questionnaires were sent to subjects in Denmark, Estonia, Iceland, Norway, and Sweden from 1999 to 2001 (response rate in women, 77%). Pregnant women, women using hormone replacement therapy, and women >45 years were excluded. Analyses included 5791 women 25 to 44 years old, of whom 961 (17%) used OCP. Logistic regression analyses included adjustment for smoking, irregular menstruation, BMI, age, type of dwelling, and center. RESULTS: Oral contraceptive pills were associated with increased risk for asthma (odds ratio, 1.42; 95% CI, 1.09-1.86), asthma with hay fever (1.48; 1.08-2.03), wheeze with shortness of breath (1.27; 1.02-1.60), hay fever (1.25; 1.06-1.48), and >/=3 asthma symptoms (1.29; 1.05 1.58). The findings were consistent between centers. The associations were present only among normal weight women (BMI 20-25 kg/m(2), asthma: 1.45; 1.02 2.05) and overweight women (BMI >25kg/m(2): 1.91; 1.20-3.02), but not among lean women (BMI <20 kg/m(2): 0.41; 0.12-1.40). Interaction between BMI and OCP in association with asthma was significant (P(interaction) < .05). CONCLUSIONS: Women using oral contraceptive pills had more asthma. This was found only in normal weight and overweight women, indicating interplay between sex hormones and metabolic status in effect on the airways. The findings originate from a cross sectional postal survey and should be interpreted with caution; it is recommended that asthma symptoms are included in clinical trials of oral contraception. PMID- 19121864 TI - Quality and lipid composition of spermatozoa in rabbits fed DHA and vitamin E rich diets. AB - The effects of fish oil (FO) and vitamin E (vE) dietary supplementation on semen quality, sperm susceptibility to lipid peroxidation, tocopherols content and fatty acid profiles were studied in rabbits. Fifty-two rabbit bucks randomly divided in four groups received a control diet and enriched diets containing either FO (1.5%, w/w), vE (200mg/kg) or both. Semen volume, concentration, motility and viability were analysed at various time-points and the lipid composition was assessed on sperm cells. The phospholipid fatty acid profile was determined: n-6 PUFA were the major fatty acids found, with a proportion of 42%, whereas the n-3 PUFA accounted for nearly 1%, mainly represented by C22:6n-3 (docosahexaenoic acid, DHA). FO supplementation produced a seven-fold increase in the content of DHA in sperm phospholipids and a comprehensive rearrangement of the phospholipid fatty acid composition, while an unexpected negative effect of feeding high level of vE on the proportion of total PUFA was found. Despite the remarkable changes observed in sperm lipid composition, semen quality parameters were not affected by the dietary treatments and the interaction between the two dietary supplements had a significant effect only on sperm concentration. An increase in semen production by ageing and a concomitant rise in sperm susceptibility to in vitro peroxidation was found. alpha- and delta-tocopherol, present in rabbit sperm in similar amount, were not affected by dietary treatment. delta-tocopherol content had a significant linear negative regression with age and showed a significant negative correlation with the susceptibility to peroxidation values. PMID- 19121865 TI - Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence. AB - Oocyte maturation is a long process during which oocytes acquire their intrinsic ability to support the subsequent stages of development in a stepwise manner, ultimately reaching activation of the embryonic genome. This process involves complex and distinct, although linked, events of nuclear and cytoplasmic maturation. Nuclear maturation mainly involves chromosomal segregation, whereas cytoplasmic maturation involves organelle reorganization and storage of mRNAs, proteins and transcription factors that act in the overall maturation process, fertilization and early embryogenesis. Thus, for didactic purposes, we subdivided cytoplasmic maturation into: (1) organelle redistribution, (2) cytoskeleton dynamics, and (3) molecular maturation. Ultrastructural analysis has shown that mitochondria, ribosomes, endoplasmic reticulum, cortical granules and the Golgi complex assume different positions during the transition from the germinal vesicle stage to metaphase II. The cytoskeletal microfilaments and microtubules present in the cytoplasm promote these movements and act on chromosome segregation. Molecular maturation consists of transcription, storage and processing of maternal mRNA, which is stored in a stable, inactive form until translational recruitment. Polyadenylation is the main mechanism that initiates protein translation and consists of the addition of adenosine residues to the 3' terminal portion of mRNA. Cell cycle regulators, proteins, cytoplasmic maturation markers and components of the enzymatic antioxidant system are mainly transcribed during this stage. Thus, the objective of this review is to focus on the cytoplasmic maturation process by analyzing the modifications in this compartment during the acquisition of meiotic competence for development. PMID- 19121866 TI - Laser surface treatment to improve mechanical properties of cast titanium. AB - OBJECTIVE: The aim of this study was to investigate the effect of laser surface treatment on the mechanical properties of cast titanium and to compare with those of the Co-Cr alloy. METHODS: Dumbbell-shaped cast specimens were prepared for commercially pure titanium (grade 2) and Co-Cr alloy. The cast titanium specimens were laser-treated on the surface using a dental Nd:YAG laser machine at 240 V and 300 V. After laser treatment, tensile testing was conducted to obtain the tensile strength, percent elongation and modulus of elasticity. The hardness depth profile was made from the cast subsurface (25 microm) to 1500 microm in depth using the cross-sections of the cast rods with the same diameter as the dumbbell. The data were statistically analyzed by ANOVA/post hoc tests (p<0.05). RESULTS: The highest tensile strength was obtained for the titanium specimens laser-treated with 300 V followed by the 240 V and the control specimens. The laser-treated titanium specimens with 300 V showed a tensile strength equivalent to the Co-Cr alloy. Although the highest modulus of elasticity was found for the specimens laser-treated with 240 V, there were no significant differences in elastic modulus among 240 V, 300 V and Co-Cr. The laser-treated groups showed significantly lower hardness at the subsurface of 25 microm and maintained their hardness until the depth of 400 microm. The hardness of the control group was very high at 25 microm depth, and dramatically decreased until the 200 microm depth. CONCLUSION: The results of tensile testing and hardness depth-profiling indicated that the laser treatment significantly improved the mechanical properties of cast titanium by improving the surface integrity of the cast surface contamination. PMID- 19121867 TI - Brief report: Direct and indirect relations of risk factors with eating behavior problems in late adolescent females. AB - This study explored correlations between risk factors and eating behavior problems in late adolescent, non-clinical females (N=301). Participants completed questionnaires for assessing eating problems, the closely associated factors of Body Mass Index (BMI) and body dissatisfaction, and a number of other risk variables that are thought to be involved in psychopathology in general, namely insecure attachment, low self-esteem, and negative affect (i.e., social anxiety and depression). Results indicated that high levels of eating problems were associated with high levels of insecure attachment, social anxiety, and depression, and with low levels of self-esteem. Further, regression analyses suggested that depression and self-esteem had a unique, direct relationship with eating problems, whereas insecure attachment and social anxiety only seemed to have indirect relations. The links between these general risk variables and eating behavior problems were independent from the specific eating problem correlates of BMI and body dissatisfaction. PMID- 19121868 TI - Staff and client perspectives on the Journey Mapping online evaluation tool in a drug court program. AB - The purpose of this study was to assess staff and client perspectives on the Internet-based Journey Mapping program evaluation tool. A drug court program was chosen for a case study research design. Six staff and 10 clients participated in interviews and observations, and also responded to a questionnaire. A staff survey provided additional data. The study revealed evidence that (1) Journey Mapping was user-friendly, saved time, promoted positive thinking, facilitated communication, increased learning, and created client voice; (2) Journey Mapping, as a program evaluation tool, supported both administrative and treatment functions of the drug court; and (3) Journey Mapping's positive orientation supplied momentum for clients' treatment progress. Journey Mapping's distinctive technology offered a viable option for including narrative methods along with quantitative methods in program evaluation. PMID- 19121869 TI - Determination of relative assay response factors for toxic chlorinated and brominated dioxins/furans using an enzyme immunoassay (EIA) and a chemically activated luciferase gene expression cell bioassay (CALUX). AB - Determination of toxic activity requires knowledge of both the concentration and toxicity to evaluate the risk for adverse human health and environmental effects. A chemically-activated luciferase gene expression cell bioassay system (CALUX) and an antibody-based method enzyme immunoassay (EIA) were used to detect the dioxin-like response of several polybrominated, polychlorinated, and polybrominated/chlorinated dibenzo-p-dioxins/furans (PBDDs/Fs, PCDDs/Fs, and PBCDDs/Fs, respectively). It has been suggested that the biological activity of the brominated and mixed bromo/chloro compounds is similar to their chlorinated analogues (measured by binding to the Ah receptor). PBDD/F, PCDD/F, and PBCDD/F laboratory standards exhibited biological activity ranging over three orders of magnitude. The highest relative potency (REP) values from CALUX analysis, when compared to 2,3,7,8-TCDD, were 2,3,7,8-TBDD at 0.99 (+/-0.07), 1,2,3,7,8-PeCDD at 0.69, and 2-Br-3,7,8-TriCDD at 0.72 (+/-0.02). Cross-reactivities were calculated using EIA for several PBDDs/Fs and PBCDDs. The highest percent cross-reactivity was found for 2,3,7,8-TBDD at 138 (+/-34%), and 2,3,7-TriBDD at 84 (+/-36%). PMID- 19121870 TI - RIP2-beta: a novel alternative mRNA splice variant of the receptor interacting protein kinase RIP2. AB - RIP2/RICK/CARDIAK is a member of the receptor interacting protein kinase (RIP) family. RIP2 promotes NF-kappaB activation as well as activation of the MAPKs JNK, ERK1/2 and p38 MAPK, thereby playing an emergent role in the innate immune response and NOD signaling. Moreover, RIP2 has been shown to interact with the CARD of caspase-1 and to induce IL-1beta maturation as well as in the induction of CD95-mediated programmed cell death by enhancing caspase-8 activity. Here, we report the identification and characterization of a novel alternative mRNA splice variant of RIP2, encoding a protein designated RIP2-beta, comprised of only a portion of the N-terminal kinase domain and lacking the intermediate region and C terminal CARD. As revealed by gene transfer experiments, these structural changes in RIP2-beta are associated with a loss of activation with respect to NF-kappaB and MAPK activation, IL-1beta secretion, and caspase-8-mediated apoptosis. In conclusion, alternative mRNA splicing may be involved in the regulation of RIP2 actions, underlying the complexity of RIP2-dependent pathways regulating stress signaling and apoptosis. PMID- 19121872 TI - Retinoid metabolism (LRAT, REH) in the liver and plasma retinoids of bullfrog, Rana catesbeiana, in relation to agricultural contamination. AB - Retinoids have been extensively studied in birds, fish and mammals where their imbalances are associated with adverse effects on growth and reproduction along with decreased embryo survival and deformities. Organochlorine compounds may alter the retinoid system but little is known about the effects of agricultural contaminants on their metabolism. In the Yamaska River project, the retinoid system in bullfrogs is monitored to investigate the possible impact of agricultural contaminants on retinoid homeostasis. Retinoids were measured in liver and plasma of male bullfrogs collected from six locations subject to increasing agricultural activity in the Yamaska River watershed. Bullfrogs living in medium and high agricultural activity areas demonstrated lower hepatic retinyl palmitate and higher hepatic retinol levels when compared to frogs associated with low contaminated sites. Changes in the concentration of hepatic esters could be related to an altered activity of REH or LRAT, enzymes respectively linked to the hydrolysis of retinyl palmitate and the esterification of retinol. A partial characterization and the analysis of liver microsomial REH and LRAT showed significantly higher hydrolysis and lower esterification activities in highly contaminated sites. Enzymatic activities seemed to be influenced by plasma DROH but not by plasma retinol. Bullfrogs from the most contaminated sites showed altered retinoic metabolism that should increase concern for frogs living in intensive agricultural areas. PMID- 19121871 TI - Depression is an early disease manifestation in lupus-prone MRL/lpr mice. AB - Many lupus patients develop neuropsychiatric manifestations, including cognitive dysfunction, depression, and anxiety. However, it is not clear if neuropsychiatric lupus is a primary disease manifestation, or is secondary to non CNS disease. We found that MRL/lpr lupus-prone mice exhibited significant depression-like behavior already at 8 weeks of age, despite normal visual working memory, locomotor coordination and social preference. Moreover, depression was significantly correlated with titers of autoantibodies against DNA, NMDA receptors and cardiolipin. Our results indicate that lupus mice develop depression and CNS dysfunction very early in the course of disease, in the absence of substantial pathology involving other target organs. PMID- 19121873 TI - The new cooperative medical scheme in China. AB - The New Cooperative Medical Scheme (NCMS) is a heavily subsidized voluntary health insurance program established in 2003 to reduce the risk of catastrophic health spending for rural residents in China. In this review, we present the current collection of knowledge available regarding the performance of NCMS on the aspects of revenue collection, risk pooling, reimbursement rules and provider payment. The available evidence suggests that NCMS has substantially improved health care access and utilization among the participants; however, it appears to have no statistically significant effect on average household out-of-pocket health spending and catastrophic expenditure risk. As NCMS is rolled out to other counties, it must be careful to generalize the findings reported in the published papers and reports, because the early pilot counties were not randomly selected and there are a lot of local adaptations. In addition, we expect that NCMS could be an important opportunity to establish some trust-based institutions in the best interest of the participants to monitor provider quality and control cost inflation. Rigorous evaluations, based on richer and latest micro-level data, could considerably strengthen the evidence base for the performance and impact of NCMS. PMID- 19121874 TI - To determine the cytotoxicity of chlorambucil and one of its nitro-derivatives, conjugated to prasterone and pregnenolone, towards eight human cancer cell-lines. AB - Four ester prodrugs derived from the bifunctional alkylating agent chlorambucil, and one of its nitro-derivatives, 3-nitrochlorambucil conjugated to prasterone and pregnenolone, were synthesized and tested for their cytotoxic activity against eight human cell lines, using the standard MTT assay. A comparison between the esters and the controls, namely chlorambucil and 3-nitrochlorambucil would suggest that all four esters possess to varying degrees, specificity towards the breast adenocarcinoma cell line (MDA-mb468) than the other seven cells' lines tested. The overall findings are encouraging since it infers that these lipophilic esters not only have the ability to traverse specific cell membranes but also exhibit cytotoxicity towards most of the cell lines tested. PMID- 19121875 TI - A multi-method study to determine the effectiveness of, and student attitudes to, online instructional videos for teaching clinical nursing skills. AB - E-learning is regularly promoted in higher education settings as a way of fostering more flexible approaches to learning. It has been argued however that the 'potential benefits of new information and communication technology instruments in education' have not been subjected to critical scrutiny (Debande, O., 2004. ICTs and the development of e-learning in Europe: the role of the public and private sectors. European Journal of Education 39 (2), 191-208, p. 192). This paper outlines a multi-method evaluation of an e-learning innovation designed to teach clinical skills to student nurses. Responding to the challenges of teaching clinical skills to large class sizes, we developed a set of instructional videos for one undergraduate skills-based module, which are now integral to the module and available online to students on a continuous basis. Evaluation suggests that students' performance outcomes are unchanged. The students view the flexible and self-management aspects of this method of learning positively, with some attitudinal differences between male and female, and mature and non-mature students. However, it is best used to complement rather than replace lecturer demonstration, lending support to a 'blended' model (Collis, B., van der Wende, M., 2002. Models of Technology and Change in Higher Education: An International Comparative Survey on The Current and Future Use of ICT in Higher Education, University of Twente, Center for Higher Education Policy Studies, The Netherlands). PMID- 19121876 TI - Whither nursing education? Possibilities, panaceas, and problems. AB - SUMMARY: The paper examines some potential problems, possibilities, and panaceas in a period where education is undergoing significant change. There are increasing class sizes and demands on clinical placements by educational institutions, and academic institutions are embracing information technology (IT) as a solution for multiple educational challenges. The paper presents possibilities and problems associated with new cohorts of nursing students entering nursing education. It also explores possibilities and problems associated with IT. Difficulties with regarding IT as a panacea for problems are considered. IT contributions to learning outcomes and reducing other systemic problems, as well as costs incurred with emphasizing IT as a teaching approach, are critically examined. The paper suggests traditional approaches to teaching (lecturing and small seminar discussions) do not preclude engagement, innovation, openness and creativity and new cohorts of students value traditional approaches. Finally, the implications for retention and recruitment of nursing educators associated with student cohort-based problems and possibilities, and costs associated with IT are considered. PMID- 19121878 TI - Elective management of the neck in oral cavity squamous carcinoma: current concepts supported by prospective studies. AB - The incidence of occult cervical metastasis in oral cavity cancer, even in early stages, is significant, necessitating elective treatment of the neck in a majority of cases. There is no method of imaging or other examination that will detect microscopic foci of metastatic disease in cervical lymph nodes. Immunohistochemical and molecular analysis of neck specimens reveals the incidence of occult metastases to be higher than revealed by light microscopy with ordinary hematoxylin and eosin staining. The neck may be treated electively by surgery or irradiation. Surgery has the advantage of permitting pathological staging of the neck, avoiding unnecessary radiation treatment and indicating cases where adjuvant therapy should be employed. As oral cavity cancer rarely metastasizes to level V, a radical or modified radical neck dissection of all five node levels is not necessary. Selective dissection of levels I-III ("supraomohyoid neck dissection") is the usual procedure of choice for elective dissection of the neck. Most of the relatively small number of isolated metastasis to level IV are from primary tumours of the tongue, which are known to produce "skip" metastases. Thus an "extended supraomohyoid neck dissection" of levels I-IV is recommended by some authors for elective treatment of the neck in tongue cancer. A number of recent prospective multi-institutional studies have demonstrated that sublevel IIB is rarely involved with isolated metastasis from oral cavity primary tumours, except from some tongue cancers. Thus it is justifiable to omit dissection of sublevel IIB in elective treatment of most cases of oral cavity cancer. Bilateral neck dissection should be performed in elective treatment of tumours involving midline structures, and in patients with ipsilateral neck metastasis. PMID- 19121879 TI - Ultrasound-guided core biopsy for investigation of cervical lymph node: chronic lymphocytic leukaemia and metastatic squamous cell carcinoma. AB - We describe a case in which ultrasound-guided fine needle core biopsy of a cervical lymph node enabled diagnosis of dual disease. Histological examination of the core biopsy confirmed unsuspected chronic lymphocytic leukaemia with an isolated focus of metastatic squamous cell carcinoma (SCC), and enabled optimal preoperative planning of treatment. Such a case is extremely unusual and provides evidence of the accurate diagnosis that can be obtained from nodal investigations using ultrasound-guided core biopsy. PMID- 19121880 TI - The role of dynamic flexion in spine injury is altered by increasing dynamic load magnitude. AB - BACKGROUND: Evidence indicates that loads and postures that an individual is exposed to alter their risk of reporting low back pain or incurring a spine injury. In vitro research indicates cyclic flexion under static compressive loads can lead to disc herniation, while repetitive compression in neutral or flexed postures leads to vertebral failure. However, no research has examined the likelihood of altering injury site (disc vs. bone) when dynamic load exposures are varied concurrently with cyclic flexion. METHODS: Fifty porcine cervical spinal units were assigned to one of five groups based on peak normalized loads of 10%, 30%, 50%, 70% and 90% of the unit's predicted tolerance. Specimens underwent passive range of motion tests to determine individualized range of motion. Once individualized loads and angles were determined, specimens were cyclically compressed and flexed based on profiles obtained from a floor to waist height lift until failure occurred or 12h elapsed. After testing specimens were dissected to identify injury site, and cumulative exposures sustained to failure were calculated. FINDINGS: Disc injury was not observed when peak loads exceeded 30% of the tolerance, while they comprised a higher percentage of the total injuries incurred when decreasing from the 30% to 10% groups. Those specimens exhibiting disc injury tolerated significantly greater: cycles to failure (9000 vs. 930, P<0.0001), cumulative compression (10872.7 vs. 1089.5MNs, P<0.001), shear (1822.1 vs. 150.6MNs, P<0.001) and angular excursion (130809.7 degrees vs. 12714.7 degrees , P<0.001). INTERPRETATION: If the spine is exposed to greater levels of load, in the presence of repetitive flexion, it is more likely to experience vertebral fracture. However, if the spine is exposed to many cycles of low peak loads, injury is more likely to occur to the intervertebral disc than to the vertebral bone or endplate. PMID- 19121881 TI - The postural response of the pelvic floor muscles during limb movements: a methodological electromyography study in parous women without lumbopelvic pain. AB - BACKGROUND: Pregnancy-related lumbopelvic pain is common. More than 30% of women have persistent pain 3 months after giving birth. There is no consensus regarding the pathology. However, coordination of muscle activity by appropriate timing and amplitude is necessary for maintaining adequate stability in the lumbopelvic area. The aim was to develop a method using surface electromyography to detect a feed-forward response in the pelvic floor muscles during limb movements performed at a comfortable speed applicable in future studies for women with lumbopelvic pain. METHODS: Ten parous women with no lumbopelvic pain in the past 12 months were included. Surface electromyographic activity was recorded from the pelvic floor muscles and unilaterally from transversus abdominis/internal oblique, rectus abdominis, erector spinae, hip adductors, rectus femoris and deltoid. The subjects performed leg lift in supine and arm lift from standing. The electromyographic onset was related to the initiation of the movement. FINDINGS: In the majority of the women the electromyographic onsets of the pelvic floor muscles occurred before the movement was initiated, regardless of whether it was a leg or an arm lift. In addition, electromyographic onsets for the other muscles, except the rectus abdominis during the arm lift, also occurred prior to the movements. INTERPRETATION: The findings suggest a feed-forward response in the pelvic floor muscles during leg and arm lifts in women who had previously given birth and were without lumbopelvic pain. Movements performed at a comfortable speed seem to be useful in order to detect such a response. PMID- 19121882 TI - Comparison of an assay for Dehalococcoides DNA and a microcosm study in predicting reductive dechlorination of chlorinated ethenes in the field. AB - The study aims to compare the detection of 16S rRNA gene of Dehalococcoides species and the microcosm study for biotransformation in predicting reductive dechlorination of chlorinated ethenes in ground water at hazardous waste sites. A total of 72 ground water samples were collected from 12 PCE or TCE contaminated sites in the United States. The samples were analyzed and used to construct microcosms in the laboratory. The results showed that the presence of Dehalococcoides DNA was well associated with dechlorination to ethene in the field. Nearly half of the wells where Dehalococcoides DNA was detected had ethene as a dechlorination end product. In comparison, for ground water samples of 16 wells where ethene was detected, ethene was produced in 11 of the corresponding microcosms. For most microcosms, during two years of incubation, dechlorination was less extensive than that observed in the field. PMID- 19121883 TI - Influence of Chironomus riparius (Diptera, Chironomidae) and Tubifex tubifex (Annelida, Oligochaeta) on oxygen uptake by sediments. Consequences of uranium contamination. AB - The diffusive oxygen uptake (DOU) of sediments inhabited by Chironomus riparius and Tubifex tubifex was investigated using a planar oxygen optode device, and complemented by measurements of bioturbation activity. Additional experiments were performed within contaminated sediments to assess the impact of uranium on these processes. After 72h, the two invertebrate species significantly increased the DOU of sediments (13-14%), and no temporal variation occurred afterwards. Within contaminated sediments, it was already 24% higher before the introduction of the organisms, suggesting that uranium modified the sediment biogeochemistry. Although the two species firstly reacted by avoidance of contaminated sediment, they finally colonized it. Their bioturbation activity was reduced but, for T. tubifex, it remained sufficient to induce a release of uranium to the water column and an increase of the DOU (53%). These results highlight the necessity of further investigations to take into account the interactions between bioturbation, microbial metabolism and pollutants. PMID- 19121884 TI - Isolation and characterization of mesotrione-degrading Bacillus sp. from soil. AB - Dissipation kinetics of mesotrione, a new triketone herbicide, sprayed on soil from Limagne (Puy-de-Dome, France) showed that the soil microflora were able to biotransform it. Bacteria from this soil were cultured in mineral salt solution supplemented with mesotrione as sole source of carbon for the isolation of mesotrione-degrading bacteria. The bacterial community structure of the enrichment cultures was analyzed by temporal temperature gradient gel electrophoresis (TTGE). The TTGE fingerprints revealed that mesotrione had an impact on bacterial community structure only at its highest concentrations and showed mesotrione-sensitive and mesotrione-adapted strains. Two adapted strains, identified as Bacillus sp. and Arthrobacter sp., were isolated by colony hybridization methods. Biodegradation assays showed that only the Bacillus sp. strain was able to completely and rapidly biotransform mesotrione. Among several metabolites formed, 2-amino-4-methylsulfonylbenzoic acid (AMBA) accumulated in the medium. Although sulcotrione has a chemical structure closely resembling that of mesotrione, the isolates were unable to degrade it. PMID- 19121885 TI - The contribution of material, psychosocial, and behavioral factors in explaining educational and occupational mortality inequalities in a nationally representative sample of South Koreans: relative and absolute perspectives. AB - The contributions of material, psychosocial, and behavioral factors in explaining socioeconomic inequalities in health have been explored in many Western studies. Most prior investigations have looked at relative abilities to explain such inequalities. In addition, little research focuses on Asian countries, despite the fact that the prevalence and socioeconomic distribution of risk factors for mortality are different there. This study examined relative and absolute abilities of material, psychosocial, and behavioral pathways to explain educational and occupational inequalities in mortality in a nationally representative sample from South Korea. The 1998 and 2001 National Health and Nutrition Examination Survey data were pooled and linked to national mortality data. Of 8366 men and women over 30 years of age, 310 died between 1999 and 2005. Nine pathway variables were examined: three material factors (income, health insurance, and car ownership status), three psychosocial factors (depression, stress, and marital status), and three behavioral factors (smoking, alcohol consumption, and physical exercise). The relative risk and relative index of inequality were used as measures of relative inequality, and risk differences and the slope index of inequality were used as measures of absolute inequality. Material factors explained a total of 29.0% of the excess in relative risk for education and 50.0% of the excess in relative risk for occupational class. Material factors explained 78.6% of the excess in absolute mortality difference for education and 41.1% for occupational class. Psychosocial factors for both education and occupational class had a relative and absolute explanatory power of less than 15%. Behavioral factors showed a relative explanatory power of about 15%, but absolute explanatory power reached 84.0% for education and 105.4% for occupational class. However, the number of deaths used to calculate the absolute explanatory power was small. Results of this study suggest that absolute socioeconomic mortality inequalities could be substantially reduced if behavioral risk factors were reduced in the whole population. PMID- 19121886 TI - Comment on Iffy et al. "The risk of shoulder dystocia related permanent fetal injury in relation to birth weight". PMID- 19121887 TI - Chronic intervillositis of unknown etiology (CIUE): relation between placental lesions and perinatal outcome. AB - OBJECTIVES: To evaluate perinatal outcome of pregnancies complicated by chronic intervillositis of unknown etiology (CIUE) and to study the relation between extent of such placental histological lesions and clinical expression. STUDY DESIGN: Descriptive and retrospective study including all cases of CIUE diagnosed between 2000 and 2006 in the university hospital of Toulouse (France). Perinatal outcome was evaluated according to the extent of placental lesions assessed by semi-quantitative graduation. RESULTS: Twenty pregnancies complicated by CIUE were included (14 patients). Three pregnancies were prematurely interrupted spontaneously during the first trimester. Perinatal outcome of the remaining 17 pregnancies beyond 22 WG was: 4 intrauterine fetal deaths, 3 terminations of pregnancy for early and severe intrauterine growth restriction (IUGR), and 10 live births (58.8%). All fetal deaths, 82.3% of pregnancies beyond 22 WG and 70% of live births were growth restricted. Severe intervillositis with massive fibrinoid deposition was associated with a severe perinatal prognosis whereas focal forms had a best evolution. The rate of recurrence was 100% in the reported cases. CONCLUSION: CIUE have a poor perinatal outcome and a high rate of recurrence. There is a relation between clinical expression and histological lesions. PMID- 19121888 TI - Dilation of the endoplasmic reticulum in beta cells due to molecular overcrowding? Kinetic simulations of extension limits and consequences on proinsulin synthesis. AB - Insulin regulates the energy homeostasis of the human body. This is synthesized in the endoplasmic reticulum (ER) of pancreatic beta cells from proinsulin. Chronic hyperglycemia increases considerably the proinsulin secretion, overcrowding the ER. Recent experimental evidence demonstrates that such states favor the proinsulin denaturation. The biophysical mechanism of this cellular dysfunction remains largely unknown. We use basic molecular principles and numerical simulations of time-dependent crowding conditions in the ER to show that crowding effects enhance the propensity of proinsulin molecules to (mis)fold in compressed, nonnative structures. Present results suggest: i) misfolding events and toxic accumulations increase dramatically if the proinsulin load exceeds 50% of the available space and ii) insufficient lag time for the relaxation of the ER between consecutive proinsulin uploads can cause irreversible alterations of folding capabilities. Present study may prove useful in generating new testable statements on circumstances leading to the development of diabetes. PMID- 19121889 TI - Exploring multifunctional agriculture. A review of conceptual approaches and prospects for an integrative transitional framework. AB - In the last decade the multifunctional agriculture (MFA) concept has emerged as a key notion in scientific and policy debates on the future of agriculture and rural development. Broadly speaking, MFA refers to the fact that agricultural activity beyond its role of producing food and fibre may also have several other functions such as renewable natural resources management, landscape and biodiversity conservation and contribution to the socio-economic viability of rural areas. The use of the concept can be traced to a number of wider societal and political transformation processes, which have influenced scientific and policy approaches in different ways amongst countries and disciplines. This paper critically discusses various existing research approaches to MFA, both from natural and social sciences. To this aim different strands of literature are classified according to their focus on specific governance mechanisms and levels of analysis into four main categories of research approaches (market regulation, land-use approaches, actor-oriented and public regulation approaches). For each category an overview of the state-of-the-art of research is given and an assessment is made of its strengths and weaknesses. The review demonstrates that the multifunctionality concept has attracted a wealth of scientific contributions, which have considerably improved our understanding of key aspects of MFA. At the same time approaches in the four categories have remained fragmented and each has limitations to understand MFA in all its complexity due to inherent constraints of applied conceptualizations and associated disciplinary backgrounds. To go beyond these limitations, we contend, new meta-level frameworks of analysis are to be developed that enable a more integrated approach. The paper concludes by presenting the main lines of an integrative, transitional framework for the study of MFA, which analyses multifunctional agriculture against the background of wider societal change processes towards sustainability and identifies a number of key elements and research challenges for this. PMID- 19121890 TI - Objective tremor registration during DBS surgery for essential tremor. AB - Essential Tremor (ET) is characterized by a 4-12-Hz postural and kinetic tremor, most commonly affecting the upper limbs. Deep brain stimulation (DBS) of the thalamus (Vim) has been found to be highly effective in severe/refractory forms of ET. Intra-operative assessment of tremor is performed using clinical methods based on patient and physician perception of tremor intensity. We present for the first time the case of a patient whose tremor was objectively monitored/quantified pre- and intra-operatively using device-based tremor registration to supplement clinical measures. We present the case of a 76-year old right-handed woman that received unilateral (left-sided) DBS of the ventrointermediate (Vim) nucleus of thalamus (Vim) for medically refractory ET. Tremor was monitored with an accelerometer-based Tremor Pen, which is part of a simple portable device (CATSYS 2000 System, Danish Product Development Ltd., DK, www.catsys.dk). The patient was asked to perform tasks for tremor evaluation before and during thalamic DBS. Tremor quantification revealed a significant improvement (34.7-fold) in the contralateral (right) limb following macro stimulation. No significant improvement was registered in the ipsilateral (non operated) side. Simple electronic tremor registration methods during DBS of the Vim nucleus of the thalamus may supplement the existing methodology that is solely based on subjective measures derived from clinical observations. PMID- 19121891 TI - PAHs soil decontamination in two steps: desorption and electrochemical treatment. AB - The presence of carcinogenic polycyclic aromatic hydrocarbons (PAHs) in soils poses a potential threat to human health if exposure levels are too high. Nevertheless, the removal of these contaminants presents a challenge to scientists and engineers. The high hydrophobic nature of PAHs enables their strong sorption onto soil or sediments. Thus, the use of surfactants could favour the release of sorbed hydrophobic organic compounds from contaminated soils. In this work, five surfactants, namely Brij 35, Tergitol NP10, Tween 20, Tween 80 and Tyloxapol, are evaluated on the desorption of PAHs [benzanthracene (BzA), fluoranthene (FLU), and pyrene (PYR), single and in mixture] from a model sample such as kaolin. In all cases, the best results were obtained when Tween 80 was employed. In order to obtain the global decontamination of PAHs, their electrochemical degradation is investigated. It is concluded that the order of increasing degradation for single compounds is BzA>FLU>PYR when they are subject to the same electrochemical treatment. In addition, there is a direct relationship between the ionization potential and the electrochemical degradation of PAH. PMID- 19121892 TI - Recovery of metals from waste printed circuit boards by a mechanical method using a water medium. AB - Research on the recycling of waste printed circuit boards (PCB) is at the forefront of environmental pollution prevention and resource recycling. To effectively crush waste PCB and to solve the problem of secondary pollution from fugitive odors and dust created during the crushing process, a wet impacting crusher was employed to achieve comminution liberation of the PCB in a water medium. The function of water in the crushing process was analyzed. When using slippery hammerheads, a rotation speed of 1470 rpm, a water flow of 6m(3)/h and a sieve plate aperture of 2.2mm, 95.87% of the crushed product was sized less than 1mm. 94.30% of the metal was in this grade of product. Using smashed material graded -1mm for further research, a Falcon concentrator was used to recover the metal from the waste PCB. Engineering considerations were the liberation degree, the distribution ratio of the metal and a way to simplify the technology. The separation mechanism for fine particles of different densities in a Falcon concentrator was analyzed in detail and the separation process in the segregation and separation zones was deduced. Also, the magnitude of centrifugal acceleration, the back flow water pressure and the feed slurry concentration, any of which might affect separation results, were studied. A recovery model was established using Design-Expert software. Separating waste PCB, crushed to -1mm, with the Falcon separator gave a concentrated product graded 92.36% metal with a recovery of 97.05%. To do this the reverse water pressure was 0.05 MPa, the speed transducer frequency was set at 30 Hz and the feed density was 20 g/l. A flow diagram illustrating the new technique of wet impact crushing followed by separation with a Falcon concentrator is provided. The technique will prevent environmental pollution from waste PCB and allow the effective recovery of resources. Water was used as the medium throughout the whole process. PMID- 19121893 TI - Highly dealuminated Y zeolite as efficient adsorbent for the hydrophobic fraction from wastewater treatment plants effluents. AB - In this work we report that highly dealuminated zeolite Y is a hydrophobic material that is able to remove selectively fatty acids and hydrocarbon compounds from the effluent of an urban wastewater treatment plant (UWTP). This adsorbent capability of zeolite Y could lead to an improved quality of UWTP effluents. Typical domestic wastewaters contain detergents, soaps and surfactants that are only partially removed in conventional UWTP. In the present work using an effluent from a UWTP located at Ribarroja del Turia (Valencia, Spain) containing 10 ppm of total organic carbon, we have been able to retain by adsorption on the dealuminated Y zeolite up to 16 and 60% of the organic matter of the effluent at pH values 7.2 and 4, respectively. Characterization of the adsorbed organic matter by Fourier transformed infrared (FT-IR), (1)H NMR and GC-MS after derivatization has shown that the zeolite adsorbs selectively the hydrophobic compounds of the effluent. PMID- 19121894 TI - Pharmaceutical residues in wastewater treatment works effluents and their impact on receiving river water. AB - Various pharmaceutical residues are being discharged from wastewater treatment works (WTW) effluents, the impact of which on river water quality is of high relevance to environmental risk assessment. The concentrations of eleven pharmaceutical compounds were determined in three WTWs in England, and the river Ouse receiving effluents from Scaynes Hill WTW. Results show that five compounds propranolol, sulfamethoxazole, carbamazepine, indomethacin and diclofenac were detected in all wastewater and river water samples, with carbamazepine showing the highest concentrations (up to 2336 ng L(-1)) in WTW influent. Different compounds were removed to different extent in the WTWs, varying from 43 to 92%, with the highest performance obtained by the WTW with tertiary treatment (sand filtration). The pharmaceutical residues from Scaynes Hill WTW were eventually discharged into the river Ouse, causing an elevation in their concentrations downstream of the outfall. This was confirmed by the good agreement between measured concentrations and those predicted by a simple dilution model. PMID- 19121895 TI - Nanocrystalline zinc oxide for the decontamination of sarin. AB - Nanocrystalline zinc oxide materials were prepared by sol-gel method and were characterized by X-ray diffraction, scanning electron microscopy, thermogravimetry, nitrogen adsorption and infrared spectroscopy techniques. The data confirmed the formation of zinc oxide materials of zincite phase with an average crystallite size of approximately 55 nm. Obtained material was tested as destructive adsorbent for the decontamination of sarin and the reaction was followed by GC-NPD and GC-MS techniques. The reaction products were characterized by GC-MS and the data explored the role of hydrolysis reaction in the detoxification of sarin. Sarin was hydrolyzed to form surface bound non-toxic phosphonate on the surface of nano-zinc oxide. The data also revealed the values of rate constant and half-life to be 4.12h(-1) and 0.16 h in the initial stages of the reaction and 0.361 h(-1) and 1.9h at the final stages of the reaction for the decontamination reaction on nanocrystalline ZnO. PMID- 19121896 TI - Effect of V2O5 on the properties of mullite ceramics synthesized from high aluminum fly ash and bauxite. AB - In this communication, high-strength mullite ceramics was prepared from bauxite and high-aluminum fly ash that is a by-product of coal combustion in thermal power plants. The effects of the doping V(2)O(5) on the bulk density, apparent porosity, bending strength and microstructure of mullite ceramics were studied in detail. It was indicated that 5-10 mol% V(2)O(5) reduced the sintering temperature by 50 degrees C. The apparent porosity and water absorption of the mullite ceramics decreased with increasing V(2)O(5) content. Mullite ceramics with bending strength as high as 108 MPa were obtained at 1500 degrees C with the addition of 10 mol% V(2)O(5). X-ray diffraction analysis suggested that the prepared ceramics was mainly in phase of mullite, and scanning electron microscope images confirmed that it mostly existed in the shape of a long parallelepiped. This research may provide a new method in utilizing the vast resources of fly-ash waste from power plants in the production of low-cost mullite-based engineering materials. PMID- 19121897 TI - Sulfuric acid baking and leaching of spent Co-Mo/Al2O3 catalyst. AB - Dissolution of metals from a pre-oxidized refinery plant spent Co-Mo/Al(2)O(3) catalyst have been tried through low temperature (200-450 degrees C) sulfuric acid baking followed by mild leaching process. Direct sulfuric acid leaching of the same sample, resulted poor Al and Mo recoveries, whereas leaching after sulfuric acid baking significantly improved the recoveries of above two metals. The pre-oxidized spent catalyst, obtained from a Korean refinery plant found to contain 40% Al, 9.92% Mo, 2.28% Co, 2.5% C and trace amount of other elements such as Fe, Ni, S and P. XRD results indicated the host matrix to be poorly crystalline gamma- Al(2)O(3). The effect of various baking parameters such as catalyst-to-acid ratio, baking temperature and baking time on percentage dissolutions of metals has been studied. It was observed that, metals dissolution increases with increase in the baking temperature up to 300 degrees C, then decreases with further increase in the baking temperature. Under optimum baking condition more than 90% Co and Mo, and 93% Al could be dissolved from the spent catalyst with the following leaching condition: H(2)SO(4)=2% (v/v), temperature=95 degrees C, time=60 min and Pulp density=5%. PMID- 19121898 TI - Enhancement strategies for Cu(II), Cr(III) and Cr(VI) remediation by a variety of seaweed species. AB - Various chemical treatments have been applied to six brown, red and green seaweed species with a view to enhancing their metal removal for Cu(II), Cr(III) and Cr(VI). Treatment with acetone resulted in the greatest enhancement for both cationic and anionic species with relatively low mass losses (15-35%), indicating its low risk to biomass operational stability. Cation binding was increased by 69%, while the total Cr removal was augmented by 15%. Cr(VI) binding was shown to be an adsorption-coupled reduction, whereby Cr(VI) was bound to the biomass surface at pH 2 and subsequently reduced to Cr(III). Acetone treatment also resulted in biomasses that were capable of converting up to 83% of Cr(VI) in solution to Cr(III). Blocking of carboxyl and amino functionalities had significant negative effects both on total Cr removal as well as percentage conversion of Cr(VI) to Cr(III). Results therefore indicated the significant role played by these moieties in metal binding to these seaweeds. Potentiometric titrations displayed agreement between the degree of esterification and the decrease in Cu(II) removal for Ulva spp. and Polysiphonia lanosa. FTIR analysis identified changes in biomass functionality and availability after chemical modification, the results of which were in agreement with metal removal studies. In conclusion, these biosorbents represent suitable candidates to replace conventional removal technologies for metal bearing wastewaters, in particular for the detoxification of hazardous Cr(VI) waste streams. PMID- 19121899 TI - Atomic force microscopy characterization of the chemical contrast of nanoscale patterns fabricated by electron beam lithography on polyethylene glycol oxide thin films. AB - The present paper shows that atomic force microscopy (AFM) imaging of friction force and phase lag in ambient air can be used to characterize the chemical contrast induced by electron beam (EB) irradiation on polyethylene glycol oxide (PEO) surface. Time-of-flight secondary emission mass spectroscopy measurements showed that the EB irradiation generates chemical contrast on PEO surface by decreasing the ether bond density. The AFM measurements showed smaller phase lag and lower friction and adhesive forces on the EB irradiated PEO surface, as compared to the non-irradiated PEO surface. While the chemical contrast in friction force had a linear dependence on the EB irradiation dose, the dependence of the chemical contrast in the phase lag was strongly non-linear. As the friction and adhesive forces depended on the AFM probe hydrophilicity and air humidity, the contrast in friction and adhesive forces is ascribed to different capillary condensation of ambient water vapour at the AFM tip contact with the EB irradiated and non-irradiated PEO surfaces, respectively. PMID- 19121900 TI - Corpus luteum size and plasma progesterone concentration in cows. AB - It is often assumed that a larger corpus luteum will produce more progesterone and generate higher circulating plasma concentrations. The aim of the study was to determine whether the size of the corpus luteum does actually determine circulating plasma progesterone concentrations. Data were collated from a number of studies on various aspects of luteal function in non-lactating dairy cows to allow comparisons to be made between corpus luteum weight and plasma progesterone concentration across the luteal phase. In these studies oestrous cycles had been synchronised and animals slaughtered on day 5, day 8 or day 16 following oestrus. Both corpus luteum weight and plasma progesterone concentration increased between day 5 and day 8. Plasma progesterone concentration but not luteal weight also increased between day 8 and day 16. On day 5 there was a strong relationship between corpus luteum weight and plasma progesterone (R(2)=0.64; P<0.001). However, no such relationship was present on day 8 or day 16. These results indicate that while during the early stage of corpus luteum development a relationship between size and progesterone is present, by day 8 of the cycle, the size of the corpus luteum is no longer of importance in determining circulating progesterone concentrations. PMID- 19121901 TI - Hysteroscopy for asymptomatic postmenopausal women with sonographically thickened endometrium. AB - Endometrial carcinoma is the most common genital cancer in women. While patients usually present with vaginal bleeding, in 10-20% this characteristic symptom is absent. Endometrial thickness (double layer) is measured by transvaginal sonography and thickening indicates an increased risk of malignancy or other pathology (hyperplasia or polyps). OBJECTIVE: We sought to correlate hysteroscopic and pathological findings in asymptomatic postmenopausal women with sonographically thickened endometrium (>6mm). STUDY DESIGN: A prospective observational study in a university hospital of 304 postmenopausal women referred between 1996 and 2006 because of a sonographically thickened endometrium in the absence of abnormal bleeding, who underwent continuous flow hysteroscopy (4.5mm Storz hysteroscope) and fractionated curettage of the uterine cervix and corpus (D & C) in addition to vaginal sonography (5MHz probe). RESULTS: The mean age of the women was 64.8 (range 57.7-71.9) years. Average endometrial thickness measured by ultrasound was 12mm+/-6.7mm. Hysteroscopy suggested the presence of endometrial polyps in 226 women (74.3%), simple endometrial hyperplasia in 34 (11.2%), atrophic endometrium in 18 (5.9%), complex endometrial hyperplasia in 2 (0.7%), atypical hyperplasia in 3 (1%) and leiomyoma in 9 (3.0%). In 12 women (3.9%), the hysteroscopic appearance suggested malignancy and histology revealed endometrial adenocarcinoma. All hysteroscopic results were confirmed by histological examination. CONCLUSION: Hysteroscopy represents an easy, safe and effective method for the investigation of asymptomatic women with a thickened endometrium found with transvaginal ultrasound. The commonest pathology was endometrial polyps. PMID- 19121902 TI - Recurrence of a laryngeal spindle cell sarcoma with a transformation into a higher grade of malignancy. AB - Primary malignant mesenchymal neoplasms of the larynx are rare. Sarcomas of the larynx account for <1% of all malignant laryngeal mesenchymal neoplasms. This report examines a case of a recurring laryngeal, initial benign-appearing mesenchymal tumour, which first changed its clinical phenotype without any histological signs of malignancy and later also its histological appearance with signs of malignancy. Finally, it even underwent a transformation into a higher grade of malignancy. In addition to this, the difficulties of allocating this tumour to the correct sarcoma subentity are shown. PMID- 19121903 TI - Demonstration of pulmonary perfusion heterogeneity induced by gravity and lung inflation using arterial spin labeling. AB - OBJECTIVE: To evaluate the effect of gravity and lung inflation on pulmonary perfusion heterogeneity in human lung using an arterial spin labeling (ASL) sequence called flow sensitive alternating inversion recovery (FAIR). MATERIALS AND METHODS: Magnetic resonance imaging of lung perfusion using arterial spin labeling sequence was performed in supine position in ten healthy volunteers on a 1.5T whole body scanner (GE Healthcare). Five coronal slices at an interval of 3cm from dorsal to ventral (labeled as P3, P6, P9, P12, P15, sequently) were obtained when the volunteers performed breath holding on end expiration and the relative pulmonary blood flow (rPBF) was measured. Then, another coronal perfusion-weighted image of P3 slice was obtained on end inspiration. Tagging efficiency of pulmonary parenchyma with IR (DeltaSI), rPBF and area of the P3 slice were analyzed. RESULTS: (1) Along the direction of gravity, a gradient was visually perceived as a vertical increase in rPBF. There were significant statistic differences in rPBF between any two coronal planes except that between P12 and P15. In supine position, regression coefficients of right and left lung were -4.98 and -5.16, respectively. This means that rPBF decreased 4.98 (right) and 5.16 (left) for each centimeter above the dorsal. No statistical difference was seen between ROIs placed along iso-gravitational plane. (2) For a same slice, there were significant statistic differences in DeltaSI, rPBF and area at different respiratory phases (P<0.05). Greater DeltaSI and more perfusion were observed on end expiration than on end inspiration. The area was larger on end inspiration than on end expiration. CONCLUSION: Both gravity and respiratory phase are important determinants of pulmonary perfusion heterogeneity. FAIR is sensitive to demonstrate gravity- and respiratory phase-dependent differences in lung perfusion. Positioning the patient so that the area of interest is down gravity and asking patient to hold breath on end expiration may help in detection of perfusion defects. PMID- 19121904 TI - CT enteroclysis in small bowel Crohn's disease. AB - The diagnostic evaluation of small bowel Crohn's disease has changed dramatically over the last decade. The introduction of wireless capsule endoscopy, double balloon endoscopy and the introduction of newer therapeutic agents have changed the role of imaging in the small bowel. Additionally, advances in multidetector CT technology have further changed how radiologic investigations are utilized in the diagnosis and management of small bowel Crohn's disease. This article describes how we perform CT enteroclysis in the investigation of small bowel Crohn's disease and discusses the role of CT enteroclysis in the current management of small bowel Crohn's disease. PMID- 19121905 TI - Respiratory gated multidetector computed tomography: applicable for diagnostic abdominal imaging? AB - PURPOSE: To evaluate the ability and accuracy of a respiratory gated technique used with contrast enhanced MDCT of the upper abdomen with focus on diagnostic image quality and depiction of organs and major vessels. MATERIALS AND METHODS: Forty-five adult patients who were referred to our institution for follow-up dynamic contrast enhanced abdominal CT imaging were included in this study. Respiratory gated CT scans were performed with the use of a dedicated hardware. A multiphasic CT scan was performed for each patient. Respiratory gated images were obtained between early arterial and portal venous phases during free breathing. Images of respiratory gated (RG) and breathhold (BH) phases were compared qualitatively and quantitatively by two radiologists. Definitive statistical methods were used for evaluating the scoring data, while Mann Whitney U test was used for comparison. Statistical significance was accepted for p values <0.05. RESULTS: Statistical significant difference was found for comparison of scores regarding luminal opacification and contoural integrity of intrahepatic vascular structures with scores of RG scans rated poor to moderate (e.g. 2.86+/-1.07 for luminal opacification of intrahepatic portal veins as well as border detectability) in comparison to scores of BH scans rated good to excellent (e.g. 1.37+/-1.31 for luminal opacification, 1.35+/-1.28 for border detectability of intrahepatic portal veins, p<0.001). Furthermore, statistical significant differences were found for general image noise levels (p<0.001). CONCLUSIONS: Further technical advances of RG technique could enable routine use of this technique for selected patient groups. PMID- 19121906 TI - US evaluation of patients affected by IBD: how to do it, methods and findings. AB - Intestinal US has to be performed very accurately, to obtain an exploration of both small bowel and cholical loops that will be the most complete that is possible; so, this technique requires adequately trained operators. Convex and linear probes with frequency between 3.5 and 7.5 MHz are used: the first ones for the panoramic evaluation and to study the middle-distal sigma and rectum; the second ones to perform a detailed examination of the bowel wall. US allows to evaluate not only the thickness and structure of the bowel wall, but also the content and peristalsis of the loops, their compressibility and movability, the perivisceral spaces and the abdominal organs. In Crohn's disease, US, completed everytime by the color-power-Doppler, shows wall thickening until 20 mm and above, with multi-stratified structure that could be regular, or more and more altered until the hypoechogenic view; the bowel vascularisation could be absent or less or more increased, due to the stadium of the disease itself. Furthermore, US allows to demonstrate the presence of stenosis and various other complications (abscesses inside or outside the walls, fistulas, involvement of other organs, free fluid collections); the use of II generation US contrast media could afford information about the activity of the disease. US has a very important role in the follow-up of patients with diagnosis of Crohn's disease, to monitor the response to the medical therapy and to discover complications; US must be performed as first in subjects with abdominal pain and diarrhea, to select the ones that need more invasive examinations. PMID- 19121907 TI - Application of FT-based MMSE deconvolution method for cerebral blood flow measurement in patients with leukoaraiosis. AB - INTRODUCTION: The bolus-tracking (BT) technique is the most popular perfusion weighted (PW) dynamic susceptibility contrast MRI method used for estimating cerebral blood flow (CBF), cerebral blood volume and mean transit time. The BT technique uses a convolution model that establishes the input-output relationship between blood flow and the vascular tracer concentration. Singular value decomposition (SVD)- and Fourier transform (FT)-based deconvolution methods are popular and widely used for estimating PW MRI parameters. However, from the published literature, it appears that SVD is more widely accepted than other methods. In a previous article, an FT-based minimum mean-squared error (MMSE) technique was proposed and simulation experiments were performed to compare it with the well-established circular SVD (oSVD) method. In this study, the FT-based MMSE method has been used to estimate relative CBF (rCBF) in 13 patients with white matter lesions (WMLs) (leukoaraiosis), and results are compared with the widely used oSVD method. MATERIALS AND METHODS: Thirteen patients with leukoaraiosis were imaged on a 1.5-T Siemens whole-body scanner. After acquiring the localizer and structural scans consisting of FLAIR (fluid attenuated with inversion recovery), T(1)-weighted and T(2)-weighted images, perfusion study was implemented as part of the MRI protocol. For each patient and method, two values were calculated: (a) rCBF for normal white matter (NWM) ROI, obtained by dividing the average CBF value in NWM ROI with average CBF in gray matter (GM) ROI, and (b) rCBF for WML ROI, obtained by dividing the average CBF value in WML ROI with average CBF in GM ROI. Results for the two deconvolution methods were computed. RESULTS AND DISCUSSION: A significant (P<.05) decrease in estimated rCBF was observed in the WML in all the patients using the MMSE method, while for the oSVD method, the decrease was observed in all but one patient. Initial results suggest that the MMSE method is comparable to the oSVD method for estimating rCBF in NMW while it may be better than oSVD for estimating rCBF in lesions of low flow. Studies involving a larger patient population may be required to further validate the findings of this work. PMID- 19121908 TI - Early contrast enhancement of the liver: exact description of subphases using MRI. AB - PURPOSE: The purpose of this study was to describe the subphases of early post contrast enhancement of the liver, using vessel enhancement patterns, and correlate these findings with enhancement patterns of abdominal organs. MATERIALS AND METHODS: A total of 114 patients who underwent gadolinium-enhanced abdominal magnetic resonance imaging examinations constituted the final study group, of which 56 were women (age range, 3-94 years; mean, 50 years) and 58 were men (age range, 6-85 years; mean, 54 years). Early post-contrast sequences in all patients were evaluated retrospectively by two reviewers for the determination of the presence of contrast enhancement in predetermined major vessels of the abdomen and qualitative and quantitative extent of enhancement of the renal cortex, spleen, pancreas and liver. Based on the overall findings, subphases of early contrast enhancement of the liver were described and quantitative extent of enhancement of organs was correlated with subphases of early contrast enhancement of the liver. Mann-Whitney U test and one-way unbalanced analysis of variance tests were used for the comparisons. RESULTS: Early hepatic arterial phase was observed in 14/114 patients, mid-hepatic arterial phase in 23/114 patients, late hepatic arterial phase in 33/114 patients, splenic vein only hepatic arterial dominant phase in 20/114 patients and hepatic arterial dominant phase in 24/114 patients. There was an overall association between the subphases of enhancement and the quantitative extent of enhancement for all studied organs (P<.0001). CONCLUSION: The evaluation of vessel and organ enhancement patterns has allowed the characterization of five different subphases in early post-contrast enhancement of the liver. The quantitative extent of enhancement of abdominal organs also demonstrated significant correlation with these five subphases. PMID- 19121909 TI - Simultaneous determination of tetrahydropalmatine, protopine, and palmatine in rat plasma by LC-ESI-MS and its application to a pharmacokinetic study. AB - A sensitive and specific liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method has been developed and validated for the simultaneous quantification of tetrahydropalmatine, protopine and palmatine in rat plasma using phenacetin as the internal standard (IS). Two hundred microliters plasma samples were extracted by dichloromethane under a strong basic condition. The analytes were separated by a C18 column and detected with a single quadrupole mass spectrometer. The used mobile phase was acetonitrile-water (40:60, v/v) containing 5mM ammonium acetate and 0.2% glacial acetic acid. Detection was carried out by positive electrospray ionization in selected ion reaction (SIR) mode at m/z 356.6 for tetrahydropalmatine, 354.6 for protopine, 352.6 for palmatine and 180.4 for the IS, respectively. The method was validated over the concentration range of 1.00-500ngmL(-1) and the lower limit of quantification (LLOQ) was 1.00ngmL(-1) for all three analytes. The intra- and inter-day precision values were less than 9% relative standard deviation (R.S.D.), and the relative error ranged from -7.4 to 4.8%. The extraction recoveries were on average 91.42% for tetrahydropalmatine, 84.75% for protopine, 57.26% for palmatine, and 83.18% for IS. The validated method was successfully applied to a pharmacokinetic study of tetrahydropalmatine, protopine and palmatine in rats after oral administration of Rhizoma Corydalis Decumbentis extract. PMID- 19121910 TI - Sensitive voltammetric determination of rutin at an in situ plated lead film electrode. AB - A highly sensitive sensor for the determination of rutin by adsorptive stripping voltammetry was described. It consists of a lead film modified glassy carbon electrode (LF/GCE). In the proposed procedure rutin was accumulated by adsorption on the lead film electrode and then during the stripping step a lead film and the accumulated rutin were oxidised. The electrochemical behaviour of rutin at the lead film electrode was investigated by square-wave voltammetry. In the optimal conditions the anodic peak currents (measured by square-wave voltammetry) increased linearly with the concentration of rutin in the range of 5 x 10(-10) to 1 x 10(-8)mol L(-1). The detection limit for rutin following 30s of accumulation time was equal to 2.5 x 10(-10) mol L(-1). The method was successfully applied to the determination of rutin content in the tablets without previous separation. PMID- 19121911 TI - Quantitative determination of penicillin V and amoxicillin in feed samples by pressurised liquid extraction and liquid chromatography with ultraviolet detection. AB - A rapid and simple method is proposed for the routine determination of amoxicillin (AMOX) and penicillin V (PENV) in swine feedingstuffs. The method is based on pressurised liquid extraction (PLE) followed by high performance liquid chromatography with ultraviolet detection (PLE-HPLC-UV) for antibiotic analysis. Parameters affecting PLE procedure, such as temperature, solvent composition, number of extraction cycles and sample cell size, were evaluated in order to achieve the highest extraction efficiency. The optimised method employed 11mL extraction cells, acetonitrile-water mixtures (25:75, v/v) for AMOX and (50:50, v/v) for PENV, as extraction solvent, 102.07atm of extraction pressure, 50 degrees C of extraction temperature, 5min of static time and 60% flush volume of the cell size. Extracts were filtered and directly analysed by HPLC-DAD/UV without further clean-up. Mean recovery rates for feed samples fortified with 200 500mgkg(-1) of both antibiotics were 86% for AMOX (RSD< or =6%) and 95% for PENV (RSD< or =3%). The method was successfully applied to the analysis of a commercial medicated swine feedingstuff, and the results were in good agreement with those obtained using mechanical shaking or ultrasonic extraction combined with solid phase extraction (UE-SPE), previously applied in the literature for feed analysis. The extraction efficiencies were evaluated by statistical comparison (analysis of variance, ANOVA-single factor) of the results obtained using the different extraction methods. Compared to the alternative techniques, PLE offers several practical advantages: easy to perform, fast, savings in solvent volume and in time, all steps are fully automated and further clean-up is not necessary for penicillin analysis. PMID- 19121912 TI - Characterization of membrane protein reconstitution in LUVs of different lipid composition by fluorescence anisotropy. AB - A major requirement to perform structural studies with membrane proteins is not only to define efficient reconstitution protocols, that assure a high incorporation degree in preformed liposomes, but also a protein directionality and topology that mimics its in vivo conditions. For this kind of studies, protein reconstitution in membranes systems via a detergent-mediated pathway is usually successfully adopted, since detergents are generally used in the initial isolation and purification of membrane proteins. In this study we report the reconstitution of OmpF in preformed DMPC and E. coli liposomes using two different techniques for detergent removal: (1) exclusion chromatography and (2) incubation with detergent-adsorbing beads. The incorporation degree was determined by bicinchoninic acid assay and fluorescence anisotropy was used to determine OmpF effect on the structural order of membrane lipids. These results show that protein insertion in membranes depends both on the technique used to remove detergent and on the lipids used to prepare the liposomes. Furthermore, it is possible to state that although the insertion is directly related to the size distributions of proteoliposomes, it could be efficiently recognized by steady state fluorescence anisotropy. This technique, more popular among cell biologists, can be a very practical and straightforward alternative to DLS to confirm membrane protein insertion. PMID- 19121913 TI - Determination of suxamethonium in a pharmaceutical formulation by capillary electrophoresis with contactless conductivity detection (CE-C(4)D). AB - A simple method based on capillary electrophoresis with a capacitively coupled contactless conductivity detector (CE-C(4)D) was developed for the determination of suxamethonium (SUX) in a pharmaceutical formulation. A hydro-organic mixture, consisting of 100mM Tris-acetate buffer at pH 4.2 and acetonitrile (90:10, v/v), was selected as background electrolyte (BGE). The applied voltage was 30kV, and the sample injection was performed in the hydrodynamic mode. All analyses were carried out in a fused silica capillary with an internal diameter of 50 microm and a total length of 64.5cm. Under these conditions, a complete separation between SUX, sodium ions and the main degradation products (choline) was achieved in less than 4min. The presence of acetonitrile in the BGE allowed a reduction of SUX adsorption on the capillary wall. The CE-C(4)D method was validated, and trueness values between 98.8% and 101.1% were obtained with repeatability and intermediate precision values of 0.7-1.3% and 1.2-1.6%, respectively. Therefore, this method was found appropriate for controlling pharmaceutical formulations containing suxamethonium and degradation products. PMID- 19121914 TI - Emergency department security programs, community crime, and employee assaults. AB - BACKGROUND: Violence against health care workers is a serious occupational health hazard, especially for emergency department (ED) employees. A significant degree of variability in security programs among hospital EDs is present in part due to the absence of federal legislation requiring baseline security features. Nationally, only voluntary guidelines from the Occupational Safety and Health Administration (OSHA) for the protection of health care workers exist. OBJECTIVES: The purpose of this study was to examine ED security programs and employee assault rates among EDs with different financial resources, size, and background community crime rates. METHODS: This cross-sectional survey was conducted among large and small hospitals located in communities with low or high rates of community crime. Hospital financial data were collected through the state health department, and employee assault data were abstracted from hospital OSHA logs. Comparisons were made using a chi-squared or Wilcoxon test. RESULTS: Small hospitals located in towns with low community crime rates implemented the fewest security program features despite having the second highest rate of assault-related OSHA-recordable injuries among ED employees (0.66 per 100,000 staff hours). CONCLUSION: Due to the highly stressful workplace characteristics of EDs, the risk of employee assault is universal among all hospital sizes in all types of communities. PMID- 19121915 TI - Working memory and the design of health materials: a cognitive factors perspective. AB - OBJECTIVE: Working memory and other supportive cognitive processes involved in learning are reviewed in the context of developing patient education materials. We specifically focus on the impact of certain design factors such as text format and syntax, the inclusion of images, and the choice of modality on individuals' ability to understand and remember health information. METHODS: A selective review of relevant cognitive and learning theories is discussed with regard to their potential impact on the optimal design of health materials. RESULTS: Working memory is measured as an individual's capacity to hold and manipulate information in active consciousness. It is limited by necessity, and well designed health materials can effectively minimize extraneous cognitive demands placed on individuals, making working memory resources more available to better process content-related information. CONCLUSIONS: Further research is needed to evaluate specific design principles and identify ideal uses of print versus video based forms of communication for conveying information. PRACTICE IMPLICATIONS: The process of developing health materials should account for the cognitive demands that extrinsic factors such as modality place on patients. PMID- 19121916 TI - Prognostic influence of immunohistochemically detected lymph node micrometastasis and histological subtype in pN0 oesophageal cancer. AB - INTRODUCTION: Differences in frequency and clinical impact of lymph node micrometastasis between histological subtypes of oesophageal cancer have not been determined. METHODS: 1204 lymph nodes from 32 squamous cell carcinomas and 54 adenocarcinomas with complete resection and pN0 status were re-evaluated using a serial sectioning protocol including immunohistochemistry. Intra-nodal tumour cells were classified as micrometastases (0.2-2 mm) or isolated tumour cells (<0.2 mm). RESULTS: There was no significant difference in the frequency of micrometastases between adenocarcinoma and squamous cell carcinoma (11.3% vs. 3.1%, p=n.s.). In the squamous cell carcinoma group, Kaplan-Meier curves showed a significantly prolonged 5-year survival (p=0.02) and disease free interval (p<0.01) for immunohistochemically node negative versus node positive patients. In patients with adenocarcinoma, no such difference (p=n.s. and p=n.s., respectively) was seen. In patients who did not undergo pre-treatment, those with adenocarcinoma had a significant 5-year survival (65% vs. 53%; p=0.03) and disease free interval (83% vs. 58%; p<0.05) advantage over those with squamous cell carcinoma. After pre-treatment, no difference between the histological subtypes was detected. Regression analysis did not reveal any factors that significantly affected overall survival in node negative patients. However, four factors did significantly influence disease free interval: pre-treatment (HR 3.3 [95% CI 1.2-9.1], p=0.02); micrometastasis (HR 5.3 [95% CI 1.4-19.7], p=0.01); UICC stage II vs. 0/I (HR 2.2 [95% CI 1.1-4.4], p=0.03); and adenocarcinoma (HR 0.3 [95% CI 0.1-0.9], p=0.03). CONCLUSION: The difference in frequency and clinical impact of immunohistochemically detected micrometastasis may indicate that adenocarcinoma and squamous cell carcinoma should not be treated as one entity. PMID- 19121917 TI - Lesion border detection in dermoscopy images. AB - BACKGROUND: Dermoscopy is one of the major imaging modalities used in the diagnosis of melanoma and other pigmented skin lesions. Due to the difficulty and subjectivity of human interpretation, computerized analysis of dermoscopy images has become an important research area. One of the most important steps in dermoscopy image analysis is the automated detection of lesion borders. METHODS: In this article, we present a systematic overview of the recent border detection methods in the literature paying particular attention to computational issues and evaluation aspects. CONCLUSION: Common problems with the existing approaches include the acquisition, size, and diagnostic distribution of the test image set, the evaluation of the results, and the inadequate description of the employed methods. Border determination by dermatologists appears to depend upon higher level knowledge, therefore it is likely that the incorporation of domain knowledge in automated methods will enable them to perform better, especially in sets of images with a variety of diagnoses. PMID- 19121918 TI - Effects of calcium sources and soluble silicate on bone metabolism and the related gene expression in mice. AB - OBJECTIVE: The effects of five calcium (Ca) sources were compared for bone biochemical and mechanical properties and the related gene expression using mice, from the viewpoint of their soluble silicon (Si) content. METHODS: Weanling male mice were fed diets containing 1% Ca supplemented with CaCO(3) as the control (CT), coral sand (CS), fossil stony coral (FSC), fish bone (FC) and eggshell (EC) powders, and 50 ppm of Si in the CT diet for 6 mo. The mRNA expressions related to bone remodeling were quantified by real-time polymerase chain reaction. RESULTS: Soluble Si content was 9.83, 7.17, 2.48, 0.29, and 0.20 ppm for the CS, FC, FSC, EC, and Ca-deficient basal diets, respectively. Si, CS, and FSC, in order, significantly increased dry and ash weights, Ca and hydroxyproline contents, and alkaline phosphatase and decreased tartrate-resistant acid phosphatase and urinary excretion of hydroxyproline compared with the CT group. Si significantly increased and FC decreased femoral strength and stiffness. In the mRNA expression related to osteoblastogenesis, Si and CS significantly increased runt-related transcription factor 2. Si, CS, and FSC, in order, significantly decreased and FC and EC increased peroxisome proliferator-activated receptor-gamma. In the mRNA expression related to osteoclastogenesis, Si and CS significantly increased and FC and EC decreased the osteoprotegerin/receptor activator of nuclear factor-kappaB ligand ratio, whereas Si and CS decreased transforming growth factor-beta. CONCLUSION: The results indicated that soluble silicate and CS, with the highest Si content among Ca sources, improved bone biochemical and mechanical properties through stimulation of gene expression related to osteoblastogenesis and suppression of that related to osteoclastogenesis. PMID- 19121919 TI - Inhibition of proliferation and induction of apoptosis by gamma-tocotrienol in human colon carcinoma HT-29 cells. AB - OBJECTIVE: gamma-Tocotrienol is a major component of the tocotrienol-rich fraction of palm oil, but there is limited evidence that it has antitumor activity. In particular, the effects of gamma-tocotrienol on human colon carcinoma cells have not been reported. To investigate the chemopreventive effects of gamma-tocotrienol on colon cancer, we examined its capacity to inhibit proliferation and induce apoptosis in HT-29 cells and explored the mechanism underlying these effects. METHODS: We cultured HT-29 cells in the presence of gamma-tocotrienol. The effect of gamma-tocotrienol on cell proliferation was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, mitotic index, and colony formation. The cell-cycle distribution was investigated by flow cytometry. We measured apoptosis by nuclear staining, transmission electron microscopy, and DNA fragmentation. Apoptosis-related proteins and the nuclear factor-kappaB p65 protein were determined by western blotting and immunofluorescence. RESULTS: gamma-Tocotrienol inhibited cell growth and arrested HT-29 cells in G(0)/G(1) phase. The 50% inhibitory concentration was 31.7 micromol/L (48 h). gamma-Tocotrienol-induced apoptosis in HT-29 cells was accompanied by downregulation of Bcl-2, upregulation of Bax, and activation of caspase-3. Furthermore, we found that gamma-tocotrienol reduced the expression level of total nuclear factor-kappaB p65 protein and inhibited its nuclear translocation. CONCLUSION: The results indicated that gamma-tocotrienol inhibits cell proliferation and induces apoptosis in HT-29 cells in a time- and dose dependent manner, and that this process is accompanied by cell-cycle arrest at G(0)/G(1), an increased Bax/Bcl-2 ratio, and activation of caspase-3. Our data also indicated that nuclear factor-kappaB p65 protein may be involved in these effects. PMID- 19121920 TI - Combined effects of saturated fat and cholesterol intakes on serum lipids: Tehran Lipid and Glucose Study. AB - OBJECTIVE: This study investigated the combined effect of saturated fat and cholesterol intake on serum lipids among Tehranian adults. METHODS: In 443 subjects >or=18 y, dietary intake was assessed. Height and weight were measured and body mass index was calculated. Serum cholesterol, triacylglycerol, high density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol levels were calculated. Cholesterol intakes >or=300 mg/d and saturated fat intakes >or=7% of total energy were defined as high intakes. Individuals were categorized into four groups based on cholesterol and saturated fat intakes. RESULTS: Subjects' mean age was 40.1 +/- 14.6 y; those in whom cholesterol and saturated fat intake was normal had significantly less energy and fat intake than those with high cholesterol and saturated fat intakes (P < 0.01). Saturated fat intake had a significant effect on serum total and HDL-C levels. Subjects with a normal saturated fat intake had significantly less serum total and HDL-C than those who had high saturated fat intake (P < 0.01 and P < 0.05, respectively). Adjusting for age, sex, and body mass index, the main effect of cholesterol intake on HDL-C was significant (P = 0.05). Mean serum HDL-C was lower in subjects who had normal cholesterol intake than in those with high cholesterol intake. CONCLUSION: These results show that cholesterol and saturated fat intakes have no combined effect on serum low-density lipoprotein cholesterol level, whereas cholesterol intake per se affects serum HDL-C level. PMID- 19121922 TI - Lycopene supplementation (passata sauce) reduces apoptosis but does not affect oxidant-responsive heme oxygenase-1 in human lymphocytes. AB - OBJECTIVE: We tested the hypothesis that lycopene supplementation reduces the expression of oxidant-responsive heme oxygenase-1 (HO-1) in basal conditions and in response to an oxidant challenge and determined whether this is temporally associated with increased cell viability. METHODS: We determined basal and stimulated ex vivo expression of HO-1 and cell viability in lymphocytes from volunteers after lycopene supplementation. Twenty-four healthy young men on a low lycopene diet consumed 1) 170 g of passata sauce with butter or 2) butter alone for 3 wk in a randomized crossover design. RESULTS: Plasma lycopene concentrations at the end of the tomato and control trials were 0.54 +/- 0.20 versus 0.20 +/- 0.15 micromol/L, respectively (P < 0.05). There was a significant increase in the proportion of live cells (91 +/- 5% versus 87 +/- 9%) and a corresponding reduction in apoptosis (6 +/- 4% versus 11 +/- 9%) in untreated lymphocytes after supplementation (P < 0.05), with no effect on cell viability in response to hydrogen peroxide treatment. HO-1 protein expression in basal conditions and induction of HO-1 after hydrogen peroxide treatment was not different between trials. CONCLUSION: Lycopene supplementation did not affect basal oxidative stress or susceptibility to oxidant-induced stress as indicated by the expression of the oxidant-responsive protein HO-1 and cell viability in response to hydrogen peroxide treatment. However, lycopene supplementation significantly reduced apoptosis in freshly harvested untreated lymphocytes. We conclude that this was not through an oxidant-mediated mechanism because of the lack of an effect on oxidant-responsive HO-1. PMID- 19121921 TI - Regulatory effects of a fermented food concentrate on immune function parameters in healthy volunteers. AB - OBJECTIVE: Nutrition is known to influence the immune system and can thereby modulate resistance to infection. The objective of this clinical trial was to assess the influence of a cascade-fermented food consisting of fruits, nuts, and vegetables rich in polyphenols (Regulat) on the immune system in healthy volunteers. METHODS: The clinical trial was double-blinded and placebo controlled. In total, 48 healthy men 20-48 y of age with a body mass index of 20 28 kg/m(2) were enrolled in the clinical trial. The group was characterized according to lifestyle parameters and only men with regular low to moderate intake of fruit and vegetables were enrolled. The intervention lasted for a period of 4 wk. Volunteers received Regulat twice daily or a placebo product (essence of vinegar). RESULTS: The intake of Regulat significantly enhanced intracellular glutathione content in lymphocytes (P < 0.05), monocytes (P < 0.05), and natural killer cells (P < 0.01). Furthermore, activation of natural killer cell cytotoxicity in response to interleukin-2 stimulation (P < 0.05), a reduction of total lipid peroxidation, and a reduction of soluble vascular cell adhesion molecule-1 (P < 0.01) and soluble intercellular adhesion molecule-1 (P < 0.05) as inflammatory blood markers were found in the Regulat but not in the placebo group. CONCLUSION: In summary, the results from this intervention study demonstrate promising physiologic effects of immune regulation on the innate immune system and antioxidative and anti-inflammatory parameters after Regulat supplementation. However, these promising results need to be confirmed in more volunteers with a more prolonged application to ensure significant beneficial effects of Regulat in the general population. PMID- 19121923 TI - Bone formation in trabecular bone cell seeded scaffolds used for reconstruction of the rat mandible. AB - This study tested whether different in vitro cultivation techniques for tissue engineered scaffolds seeded with human trabecular bone cells affect in vivo bone formation when implanted into critical-size defects in rat mandibles. Human trabecular cells were isolated and seeded into three types of scaffolds (porous CaCO(3), mineralized collagen, porous tricalcium phosphate). Four in vitro groups were produced: empty control scaffolds incubated with cell culture medium for 24 h; scaffolds seeded with trabecular bone cells, cultivated under static conditions for 24 h; scaffolds seeded with trabecular bone cells, cultivated for 14 days under static conditions; scaffolds seeded with trabecular bone cells, cultivated for 14 days in a continuous flow perfusion bioreactor. The scaffolds were implanted press fit into non-healing defects, 5 mm diameter, in rat mandibles. After 6 weeks the presence of human cells was assessed; none were detected. Histomorphometric evaluation showed that neither seeding human trabecular bone cells nor the culturing technique increased the amount of early bone formation compared with the level provided by osteoconductive bone ingrowth from the defect edges. It is concluded that human bone marrow stroma cells in tissue-engineered scaffolds and associated in vitro technology are difficult to test in the mandible in animal models. PMID- 19121924 TI - Dissociation of epileptic and inflammatory activity in Rasmussen Encephalitis. AB - SUMMARY: Multimodal imaging was performed in Rasmussen Encephalitis (RE) during episodes of complex-partial and focal motor status epilepticus including independent component analysis of BOLD-fMRI, arterial spin labeling perfusion imaging and diffusion tensor imaging. The active epileptic network and topographically independent brain areas showed regional hyperperfusion and progressive atrophy. The results suggest that hyperperfusion outside of the epileptic network represent active inflammation in RE and the imaging protocol presented here, allows assessing thereby the disease activity non-invasively. PMID- 19121925 TI - Comparative evaluation of bioactivity of crystalline trypsin for drying by Fourier-transformed infrared spectroscopy. AB - The purpose of this study was to evaluate the enzymatic stability of colloidal trypsin powder during heating in a solid-state by using Fourier transform infrared (FT-IR) spectra with chemoinformatics and generalized two-dimensional (2D) correlation spectroscopy. Colloidal crystalline trypsin powders were heated using differential scanning calorimetry. The enzymatic activity of trypsin was assayed by the kinetic degradation method. Spectra of 10 calibration sample sets were recorded three times with a FT-IR spectrometer. The maximum intensity at 1634cm(-1) of FT-IR spectra and enzymatic activity of trypsin decreased as the temperature increased. The FT-IR spectra of trypsin samples were analyzed by a principal component regression analysis (PCR). A plot of the calibration data obtained was made between the actual and predicted trypsin activity based on a two-component model with gamma(2)=0.962. On the other hand, a 2D method was applied to FT-IR spectra of heat-treated trypsin. The result was consistent with that of the chemoinformetrical method. The results for deactivation of colloidal trypsin powder by heat-treatment indicated that nano-structure of crystalline trypsin changed by heating reflecting that the beta-sheet was mainly transformed, since the peak at 1634cm(-1) decreased with dehydration. The FT-IR chemoinformetrical method allows for a solid-state quantitative analysis of the bioactivity of the bulk powder of trypsin during drying. PMID- 19121926 TI - Breast radiotherapy: considerations in older patients. AB - With an ageing population, the number of older women with breast cancer eligible for adjuvant irradiation after breast conserving surgery and mastectomy is rising. There is a dearth of level 1 data on the effect of adjuvant irradiation on local control, quality of life and survival. In large part this reflects the exclusion of patients over the age of 70 years from randomised trials. The prevention of local recurrence may reduce the risks of dissemination. However, older women with early breast cancer and a life expectancy of less than 5 years are unlikely to derive a survival benefit from adjuvant radiotherapy. Rates of access of older patients to adjuvant irradiation are lower than for younger patients. Physician and patient bias and co-morbidities are contributory factors. There are also competing risks of mortality from co-morbidities, particularly in women over the age of 80 years. Postoperative radiotherapy after breast conserving surgery does not seem to compromise overall quality of life of older patients. Although the absolute reduction in local recurrence from adjuvant radiotherapy is modest in lower risk older patients after breast conserving surgery and adjuvant systemic therapy, there has to date been no group of fitter old patients defined from whom radiotherapy can be reasonably omitted. Guidelines for postmastectomy radiotherapy should not differ from younger patients. Adequately powered randomised trials are needed to assess the effect of adjuvant irradiation in older patients on outcomes after breast conserving surgery and mastectomy to provide a more robust basis for evidence-based radiotherapy practice. PMID- 19121928 TI - Understanding post 9/11 drug control policy and politics in Central Asia. AB - This paper exposes contemporary drug policy challenges in Central Asia by focusing on a single point in the history of drug control, in a single region of the global war against drugs and terrorism, and on one agency whose mission is to help make the world safer from crime, drugs and terrorism. By looking closely at the post 9/11 security-oriented donor priorities, I conclude that, in Central Asia, the rhetoric of the taking a more 'balanced approach' to drug policy is bankrupt. When enacted by the national law enforcement agencies in the Central Asian republics, the 'Drug Free' aspirational goal is driving the HIV epidemic among IDUs. The face-saving 'containment' thesis does not reflect the drug situation in this region but rather the failure to adopt an evidence-based approach. The harm reduction agenda continues to face many challenges including resistance to substitution treatment, the harm from drug treatment, from poorly designed drug prevention programmes and from repressive counter-narcotics policies and practices. PMID- 19121927 TI - Clinical, functional and biochemical changes during recovery from COPD exacerbations. AB - The pathways underlying chronic obstructive pulmonary disease exacerbations (ECOPD) remain unclear. This study describes the clinical, functional and biochemical changes during recovery from ECOPD. Thirty hospitalized patients with Anthonisen's type-I ECOPD were evaluated on days 0 (admission), 3, 10 and 40. A five-symptom score (TSS), performance status and quality of life were evaluated. Post-bronchodilator spirometry, blood gases, oxidative stress, C-reactive protein (CRP), serum amyloid-A (SAA), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and fibrinogen were also measured. Patients were classified as early- or late-recoverers, based on whether dyspnea had returned to pre exacerbation level by day 10. Most clinical, functional and biochemical parameters improved during follow-up. CRP and IL-6 levels reduced on Day 3 (p<0.05), whereas SAA on Day 10 (p<0.01). TNF-alpha was reduced on Days 3 and 10, but on Day 40 its levels returned to baseline. Fibrinogen and WBC reduced only by day 40. TSS and dyspnea were correlated inversely with FEV(1) on days 3, 10 and 40. Although late-recoverers had lower FEV(1) on admission, none of the reported measurements on admission and day 3 predicted early recovery. During recovery from ECOPD, symptomatic improvement correlates only with post-bronchodilator FEV(1) whereas systemic inflammatory burden subsidence does not correlate with clinical and functional changes. Although late-recoverers have lower FEV(1) on admission, none of the measured parameters is able to predict early symptomatic recovery. PMID- 19121929 TI - New insights for using self-assembly materials to improve the detection stability in label-free DNA-chip and immuno-sensors. AB - This paper examines reliable advancements in low-cost DNA- and immuno-chips. Capacitance detection was successfully chosen to develop label-free bio-chips. Probe immobilization was rigorously investigated in order to obtain reliable capacitance measurements. Protein probes immobilized by using usual alkanethiols or thiolated ssDNA probes directly immobilized on gold do not allow sufficient stable capacitance measurements. New alkanethiols improved with ethylene-glycol function are shown in this paper to be more suitable materials for capacitive bio chip development. Atomic Force Microscopy, Quartz Crystal Microbalance, and Capacitance Measurements were used to demonstrate that ethylene-glycol alkanethiols allow high time stability, smaller errors in detection, and improved ideal behaviour of the sensing surfaces. Measured capacitance is in the range of 8-11 nF/mm(2) for antibody layers and close to 6 nF/mm(2) for DNA probes. It is in the range of 10-12 nF/mm(2) and of 4-6 nF/mm(2) for antigen and DNA detection, respectively. The percentage error in detection is highly improved and it is in the range of 11-37% and of 0,23-0,82% for antigen and DNA, respectively. The reproducibility is also improved and it is close to 0,44% for single spot measurements on ethylene-glycol alkanethiols. A molecular theory attributing these improvements to water molecules strongly coordinated by ethylene-glycol functional groups and to solution ions not entering into probe films is finally proposed. PMID- 19121930 TI - The evolution and elaboration of vertebrate neural crest cells. AB - Vertebrate neural crest cells are embryonic neuroepithelial cells that undergo an epithelial-mesenchymal transition, migrate throughout the embryo and form a wide variety of derivatives, including peripheral neurons and glia, pigment cells, and craniofacial cartilage, bone and teeth. Neural crest cell evolution and elaboration is intimately bound up with vertebrate evolution: the most primitive living vertebrates, lampreys and hagfishes, have most but not all neural crest derivatives. A torrent of recent molecular information has changed our understanding of vertebrate phylogenetic relationships, expanded our understanding of the gene circuitry underlying neural crest development, and given interesting information on the deployment of homologues of these genes in invertebrate relatives such as ascidians and amphioxus. New molecular insights into the evolutionary origin of cartilage, as well as into the nature and evolution of the cells and genes involved in tooth and bone formation, enable tentative hypotheses to be framed for the evolution of skeletal neural crest derivatives. PMID- 19121931 TI - Dietary acrylamide intake and risk of colorectal cancer in a prospective cohort of men. AB - Acrylamide is a probable human carcinogen that causes cancer at multiple sites in animal models. However, whether dietary acrylamide intake increases the risk of colorectal cancer in humans is unclear. We examined the association between dietary acrylamide intake and colorectal cancer incidence in the Cohort of Swedish Men, a population-based prospective cohort of 45,306 men who completed a food-frequency questionnaire at enrolment in 1997. During a mean follow-up of 9.3 years, we ascertained 676 incident colorectal cancer cases. Compared with the lowest quartile of acrylamide intake (<29.6 microg/d), the multivariate rate ratios for the highest quartile (> or =41.7 microg/d) were 0.95 (95% confidence interval (CI) 0.74-1.20) for colorectal cancer, 0.97 (95% CI 0.71-1.31) for colon cancer and 0.91 (95% CI 0.62-1.34) for rectal cancer. In conclusion, this study provides no evidence that dietary acrylamide in amounts typically consumed by Swedish men is associated with risk of colorectal cancer. PMID- 19121932 TI - Screen-detected breast lesions with malignant needle core biopsy diagnoses and no malignancy identified in subsequent surgical excision specimens (potential false positive diagnosis). AB - Although breast needle core biopsy (NCB) is now a standard diagnostic procedure in the triple assessment of screen-detected breast lesions, data on the false positive diagnoses of malignancy (malignant NCB 'B5' with normal/benign surgery) are lacking. In this study, we have studied a large series of NCBs (101,440) to assess the causes and pitfalls resulting in false-positive NCB diagnoses and to evaluate their impact on patients' management in the screening service. Our results showed that of 40,395 malignant NCBs reported during the period of this study, 174 NCBs are considered as false-positives (0.43%; (95% confidence interval [CI]=0.37-0.49%)). However, on review, 165 cases (95%) were found to be the result of true removal of the whole lesion in the core with subsequent negative excision biopsy samples (true-positive NCBs). This may reflect sampling of small screen detected lesions and the use of larger core biopsies at assessment. The remaining 9 cases were considered as true false-positive cores, giving a false-positive rate of 0.02% (95% CI=0.01-0.04%). Analysis of these 9 cases showed that 8 cases, originally diagnosed as DCIS, were classified as borderline lesions or lesions of uncertain malignant potential after surgical excision. The classification and management of such borderline lesions remains controversial and diagnostic surgical excision is usually the optimum management. One case was the result of pathological misinterpretation of fat necrosis as invasive carcinoma. This was the only case that resulted in a significant over management of the patient. In conclusion, our results showed that the true false positive rate of NCB is extremely rare. Significant over-management of screen detected breast lesions as a result of false-positive NCB may be considered almost nil. PMID- 19121934 TI - What to do after winning the Nobel Prize--work harder! PMID- 19121935 TI - Surgical access to the adrenal gland: the quest for a "no visible scar" approach. AB - Surgery of the adrenal glands has various approaches and remains technically challenging. Many different techniques have been used to decrease morbidity. The most important one is probably laparoscopy which has already quite dramatically improved clinical outcome. The second one is the introduction of posterior endoscopic approach (retroperitoneoscopy), which entirely preserves peritoneal integrity. This concept seems to us to be a fundamental in surgery of the retroperitoneum. Recently, new developments have emerged in the field of abdominal surgery. The most radical concept, known as Natural Orifice Transluminal Endoscopic Surgery (NOTES), consists of the introduction of both endoscope and working instruments through natural orifices in order to eliminate skin incisions and their associated morbidities. Although transperitoneal adrenalectomy has previously been reported in a porcine model, here we report our initial experiences of extra-peritoneal transvaginal NOTES approach of the retroperitoneum, focused on the field of adrenal surgery in both porcine and cadaver models. Whether clinical applications will ultimately be derived of this concept is still not clear certainly it raises new possibilities for interesting developments. PMID- 19121936 TI - Kinetics of leather dyeing pretreated with enzymes: role of acid protease. AB - In the present investigation, kinetics of dyeing involving pretreatment with acid protease has been presented. Application of acid protease in dyeing process resulted in increased absorption and diffusion of dye into the leather matrix. Enzyme treatment at 1% concentration, 60 min duration and 50 degrees C resulted in maximum of 98% dye exhaustion and increased absorption rate constants. The final exhaustion (C(infinity)) for the best fit of CI Acid Black 194 dye has been 98.5% with K and r2 values from the modified Cegarra-Puente isotherm as 0.1033 and 0.0631. CI Acid Black 194 being a 2:1 metal complex acid dye exhibited higher absorption rate than the acid dye CI Acid Black 210. A reduction in 50% activation energy calculated from Arrhenius equation has been observed in enzyme assisted dyeing process of both the dyes that substantiates enhanced dye absorption. The absorption rate constant calculated with modified Cegarra-Puente equation confirm higher rate constants and faster kinetics for enzyme assisted dyeing process. Enzyme treated leather exhibited richness of color and shade when compared with control. The present study substantiates the essential role of enzyme pretreatment as an eco-friendly leather dyeing process. PMID- 19121933 TI - Multiple tumours in survival estimates. AB - In international comparisons of cancer registry based survival it is common practice to restrict the analysis to first primary tumours and exclude multiple cancers. The probability of correctly detecting subsequent cancers depends on the registry's running time, which results in different proportions of excluded patients and may lead to biased comparisons. We evaluated the impact on the age standardised relative survival estimates of also including multiple primary tumours. Data from 2,919,023 malignant cancers from 69 European cancer registries participating in the EUROCARE-4 collaborative study were used. A total of 183,683 multiple primary tumours were found, with an overall proportion of 6.3% over all the considered cancers, ranging from 0.4% (Naples, Italy) to 12.9% (Iceland). The proportion of multiple tumours varied greatly by type of tumour, being higher for those with high incidence and long survival (breast, prostate and colon-rectum). Five-year relative survival was lower when including patients with multiple cancers. For all cancers combined the average difference was -0.4 percentage points in women and -0.7 percentage points in men, and was greater for older registries. Inclusion of multiple tumours led to lower survival in 44 out of 45 cancer sites analysed, with the greatest differences found for larynx (-1.9%), oropharynx (-1.5%), and penis (-1.3%). Including multiple primary tumours in survival estimates for international comparison is advisable because it reduces the bias due to different observation periods, age, registration quality and completeness of registration. The general effect of inclusion is to reduce survival estimates by a variable amount depending on the proportion of multiple primaries and cancer site. PMID- 19121937 TI - Discovery and SAR of benzyl phenyl ethers as inhibitors of bacterial phenylalanyl tRNA synthetase. AB - A series of benzyl phenyl ethers (BPEs) is described that displays potent inhibition of bacterial phenylalanyl-tRNA synthetase. The synthesis, SAR, and select ADMET data are provided. PMID- 19121938 TI - Preliminary investigations on the synthesis and antitumor activity of 3(2H) furanones. AB - Two triaryl-3(2H)-furanones were synthesized and their antitumor activity was evaluated. These compounds inhibited the proliferation of DLA cell line in vitro. In vivo studies also showed that these compounds were active against tumor cell proliferation. PMID- 19121939 TI - The synthesis and structure-activity relationship of substituted N-phenyl anthranilic acid analogs as amyloid aggregation inhibitors. AB - It is believed that beta-amyloid aggregation is an important event in the development of Alzheimer's disease. In the course of our studies to identify beta amyloid aggregation inhibitors, a series of N-phenyl anthranilic acid analogs were synthesized and studied for beta-amyloid inhibition activity. The synthesis, structure-activity relationship, and in vivo activity of these analogs are discussed. PMID- 19121940 TI - Syntheses of tricyclic pyrones and pyridinones and protection of Abeta-peptide induced MC65 neuronal cell death. AB - The SbetaC gene is conditionally expressed a 99-residue carboxy terminal fragment, C99, of amyloid precursor protein in MC65 cells and causes cell death. Consequently, MC65 cell line was used to identify inhibitors of toxicity related to intracellular amyloid beta (Abeta) oligomers. Compounds that reduce the level of Abeta peptides, prevent Abeta aggregation, or eliminate existing Abeta aggregates may be used in the treatment of Alzheimer's disease (AD). Previously, we found that a tricyclic pyrone (TP) molecule, compound 1, prevents MC65 cell death and inhibits Abeta aggregation. Hence various TPs containing heterocycle at C7 side chain and a nitrogen at position 2 or 5 were synthesized and their MC65 cell protective activities evaluated. TPs containing N3'-adenine moiety such as compounds 1 and 11 are most active with EC(50) values of 0.31 and 0.35 microM, respectively. EC(50) values of tricyclic N5-analog, pyranoisoquinolinone 13, and N2-analog, pyranopyridinone 20, are 2.49 and 1.25 microM, respectively, despite the lack of adenine moiety. Further investigation of tricyclic N2- and N5-analogs is warranted. PMID- 19121942 TI - Peri-substituted hexahydro-indolones as novel, potent and selective human EP3 receptor antagonists. AB - A series of peri-substituted [4.3.0] bicyclic non-aromatic heterocycles have been identified as potent and selective hEP(3) receptor antagonists. These molecules adopt a hair-pin conformation that overlaps with the endogenous ligand PGE(2) and fits into an internally generated EP(3) pharmacophore model. Optimized compounds show good metabolic stability and improved solubility over their corresponding bicyclic aromatic analogs. PMID- 19121941 TI - Amide derivatives of ethacrynic acid: synthesis and evaluation as antagonists of Wnt/beta-catenin signaling and CLL cell survival. AB - A series of amides of ethacrynic acid was prepared and evaluated for their ability to inhibit Wnt signaling and decrease the survival of CLL cells. Several of the most potent derivatives were active in the low micromolar range. Reduction of the alpha,beta-unsaturated carbon-carbon double bond of EA abrogated both the inhibition of Wnt signaling as well as the decrease in CLL survival. Preliminary mechanism of action studies suggest that these derivatives covalently modify sulfhydryl groups present on transcription factors important for Wnt/beta-catenin signaling. PMID- 19121943 TI - Gait patterns in subjects with rheumatoid arthritis cannot be explained by reduced speed alone. AB - The aim of this study was to investigate the characteristics of gait in subjects with rheumatoid arthritis (RA) by comparing gait parameters obtained from these subjects and controls. Seventeen subjects with RA in functional class II (mean age 51.1 years, S.D. 6.2 years) and 20 controls (mean age 50.4 years, S.D. 5.3 years) were instructed to walk a straight walkway at five different self-selected speeds. Speed-dependent variables were analysed by an interpolation procedure to estimate scores at a normalized speed of 0.8m/s. At self-selected speed the RA group walked significantly slower, with a shorter step length and longer stance phase. There was no difference in cadence and step width. When controlling for speed, the RA group walked with shorter step length (p=0.04) and higher cadence (p=0.03) compared to controls, but no significant difference in stand phase and step width was found. The present study demonstrates that speed-dependent gait variables are affected when controlling for the effect of speed in subjects with RA. In further studies of gait, speed should be controlled for. PMID- 19121944 TI - Correlation of binding constants and molecular modelling of inhibitors in the active sites of aldose reductase and aldehyde reductase. AB - Aldose reductase (ALR2) belongs to the aldo-keto reductase (AKR) superfamily of enzymes, is the first enzyme involved in the polyol pathway of glucose metabolism and has been linked to the pathologies associated with diabetes. Molecular modelling studies together with binding constant measurements for the four inhibitors Tolrestat, Minalrestat, quercetin and 3,5-dichlorosalicylic acid (DCL) were used to determine the type of inhibition, and correlate inhibitor potency and binding energies of the complexes with ALR2 and the homologous aldehyde reductase (ALR1), another member of the AKR superfamily. Our results show that the four inhibitors follow either uncompetitive or non-competitive inhibition pattern of substrate reduction for ALR1 and ALR2. Overall, there is correlation between the IC(50) (concentration giving 50% inhibition) values of the inhibitors for the two enzymes and the binding energies (DeltaH) of the enzyme-inhibitor complexes. Additionally, the results agree with the detailed structural information obtained by X-ray crystallography suggesting that the difference in inhibitor binding for the two enzymes is predominantly mediated by non-conserved residues. In particular, Arg312 in ALR1 (missing in ALR2) contributes favourably to the binding of DCL through an electrostatic interaction with the inhibitor's electronegative halide atom and undergoes a conformational change upon Tolrestat binding. In ALR2, Thr113 (Tyr116 in ALR1) forms electrostatic interactions with the fluorobenzyl moiety of Minalrestat and the 3- and 4-hydroxy groups on the phenyl ring of quercetin. Our modelling studies suggest that Minalrestat's binding to ALR1 is accompanied by a conformational change including the side chain of Tyr116 to achieve the selectivity for ALR1 over ALR2. PMID- 19121945 TI - Structure-activity relationship study on polyglutamine binding peptide QBP1. AB - Aggregation and deposition of expanded polyglutamine proteins in the brain cause neurodegenerative diseases including Huntington disease. This pathogenic process is suppressed and delayed in the presence of polyglutamine binding peptide 1 (QBP1), which we previously identified as an undecapeptide binding to pathogenic polyglutamine proteins from phage display peptide libraries. In this paper, a structure-activity relationship study on QBP1 was conducted to determine the pharmacophores for inhibition of polyglutamine aggregation. Furthermore, a truncation study identified an octapeptide as the minimum structure for suppressing aggregation of polyglutamine proteins, which is equipotent to the parent undecapeptide QBP1. PMID- 19121946 TI - Novel boronated chlorin e6-based photosensitizers: synthesis, binding to albumin and antitumour efficacy. AB - Chlorins, a class of plant porphyrins, are perspective as photosensitizing agents due to light absorption in the long wavelength spectral region and deeper photodamage of tissues. Aiming at optimization of antitumour properties of chlorins, we synthesized a series of boronated derivatives of chlorin e(6) and their complexes containing Zn(II), Pd(II) or Sn(IV). The compounds were synthesized by alkylation of amino or hydroxy derivatives of chlorin e(6) with 1 trifluoromethanesulfonylmethyl-o-carborane. Chlorin e(6) 13(1)-N-{2-[N-(o carboran-1-yl)methyl]aminoethyl}amide-15(2), 17(3)-dimethyl ester (compound 5) formed complexes with serum albumin, a major porphyrin carrier. The binding constant of these complexes was approximately 4 times bigger than the respective value for the complexes of albumin with boron-free aminochlorin e(6). Compound 5 potently sensitized rat fibroblasts to illumination with monochromatic red light: >98% of cells were necrotic by 24h post-illumination with 1 microM of 5. This compound demonstrated high efficacy in photodynamic therapy of rat M-1 sarcoma. After PDT with 25mg/kg of 5 the residual tumours were significantly smaller than in animals subjected to PDT with equal concentration of boron-free aminochlorin e(6). No signs of general toxicity were detectable after PDT with 5. Thus, boronation can enhance the potency of chlorins in PDT, in particular, due to an increased binding to albumin. Our data expand the therapeutic applicability of boronated chlorins beyond boron neutron capture therapy; these agents emerge as dual efficacy photoradiosensitizers. PMID- 19121947 TI - In situ study of epitaxial growth of ZnO nanowires at the junctions of nanowall networks on zinc particles. AB - The authors show the real-time growth of ZnO nanowalls and nanowires on zinc particles via in situ observation in an environmental scanning electron microscopy. It was observed that a ZnO polycrystalline film is first deposited on zinc particles. The nanowires started to grow when the nanowalls had just formed and they grew epitaxially on the junctions of the nanowalls. The nanowalls and the nanowires grew together until the source of zinc was exhausted. The vapor solid mechanism is deemed to be the growth mechanism as it quantitatively accounts for the growth speed of the nanowalls and nanowires observed in the experiment. Cathodoluminescence reveals that the growth at low zinc concentration leads to blue emission from defects, which may be zinc vacancies. PMID- 19121948 TI - An Arabidopsis family of six acyl-CoA-binding proteins has three cytosolic members. AB - In Arabidopsis thaliana, a gene family of six members encodes acyl-CoA-binding proteins (ACBPs). These Arabidopsis ACBPs (designated ACBP1 to ACBP6) range in size from 10.4kDa to 73.1kDa and display varying affinities for acyl-CoA esters, suggesting that they have different roles in plant lipid metabolism. In contrast, only the 10-kDa ACBPs have been well-characterized from other eukaryote species. Our previous studies have revealed that ACBP1 and ACBP2 are membrane-associated proteins, while ACBP3 is extracellularly-targeted. More recently, we have reported that the remaining three members in this protein family (namely ACBP4, ACBP5 and ACBP6) are subcellularly localized to the cytosol in Arabidopsis. The subcellular localizations of ACBP4, ACBP5 and ACBP6 in the cytosol were demonstrated using a number of different approaches incorporating biochemical fractionation, confocal microscopy of transgenic Arabidopsis expressing autofluorescence-tagged fusions and immunoelectron microscopy using ACBP-specific antibodies. Our results indicate that all three ACBPs in the cytosol are potential candidates for acyl-CoA binding and trafficking in plant cells. In this review, the functional redundancy and differences among the three cytosolic ACBPs are discussed by comparison of their light-regulated expression and substrate affinities to acyl-CoA esters, and from biochemical analyses on their knockout mutants and/or overexpression in transgenic Arabidopsis. The transcriptionally light-induced ACBP4 and ACBP5, which encode the two largest forms of Arabidopsis ACBPs, bind oleoyl-CoA esters and likely transfer oleoyl-CoAs from the plastids (the site of de novo fatty acid biosynthesis) to the endoplasmic reticulum for the biosynthesis of non-plastidial membrane lipids in Arabidopsis. PMID- 19121949 TI - Report of rare bilateral nasolabial cysts. AB - PURPOSE: Nasolabial cysts are usually unilateral and are quite rare, while bilateral cysts are even rarer. PATIENT AND METHOD: Our report concerns a 48-year old female with bilateral nasolabial cysts. After many years of misdiagnosis she was finally referred to our clinic with a subnasal swelling of unknown origin. RESULT: Evaluation of the patient's medical history, clinical examination and of a previous CT scan led to the diagnosis of a nasolabial cyst, which was later confirmed by histological examination. Treatment involved the surgical excision. CONCLUSION: A complete surgical excision is recommended using a sublabial approach as the treatment of choice, although transnasal endoscopic marsupialization seems to be a simple and effective alternative. It has been shown that after successful marsupialization, the nasolabial cyst is converted to an air-containing paranasal sinus. PMID- 19121950 TI - Farnesol promotes epithelial cell defense against Candida albicans through Toll like receptor 2 expression, interleukin-6 and human beta-defensin 2 production. AB - Farnesol, a quorum-sensing molecule, regulates virulence and morphogenesis in Candida albicans and is involved in various human pathologies including oral candidiasis. Oral epithelial cells are involved in innate immunity against Candida infections via Toll-like receptors (TLRs) and inflammatory mediators. We investigated the effects of farnesol on host cells and its possible synergistic interaction with gingival epithelial cells against C. albicans infection by studying the expression of TLR2, 4 and 6. The production of IL-6, IL-8, and human beta-defensins 1 and 2 was also examined using engineered human oral mucosa tissue put in contact with various concentrations of farnesol with and without C. albicans. Our findings indicate that 24 h after contact with C. albicans, epithelial cells expressed more TLR2 than did non-infected cells. The addition of exogenous farnesol upregulated the TLR2 expression by the gingival epithelial cells in the presence or absence of C. albicans. In contrast, TLR4 was down regulated when farnesol was added to the tissue with or without C. albicans. Finally, farnesol alone was shown to have no effect on TLR6, yet in the presence of both C. albicans and farnesol, TLR6 expression was down regulated. Farnesol modulated TLR2 expression by the epithelial cells following tissue contact with C. albicans. This effect was paralleled by IL-6 but not IL-8 secretion. Farnesol's effect on innate immunity was strengthened by its capacity to increase human beta-defensin 2 production, and by the efficacy of beta-defensin against C. albicans growth. Overall results showed that exogenous farnesol promoted epithelial cell defense against C. albicans infection through the involvement of TLR2, IL-6, and human beta-defensin 2. PMID- 19121951 TI - A simple method to remove ECG artifacts from trunk muscle EMG signals. PMID- 19121952 TI - Reliability of electromyographic methods used for assessing hip and knee neuromuscular activity in females diagnosed with patellofemoral pain syndrome. AB - Patellofemoral pain syndrome (PFPS) is one of the most common, yet misunderstood, knee pathologies. PFPS is thought to result from abnormal patella tracking caused from altered neuromuscular control. Researchers have investigated neuromuscular influences from the gluteus medius (GM), vastus medialis (VM), and vastus lateralis (VL) but with inconsistent findings. A reason for these discrepancies may be from varying methodology. The purpose of this study was to determine the reliability of electromyographic (EMG) methods used to assess amplitudes and timing differences of the GM, VM, and VL in subjects with PFPS. Seven females with PFPS participated. GM, VM, and VL activity was assessed during the stance phase of a stair descent task on two separate occasions. Amplitudes during the different intervals of stance were recorded and expressed as a percent of each muscle's maximum voluntary isometric contraction. Muscle onsets at the beginning of stair descent were also determined. VM-GM, VL-GM, and VL-VM onset timing differences were quantified. Intraclass correlation coefficients (ICCs) and standard errors of measurement (SEMs) were calculated to assess between-day reliability. Most EMG measures had acceptable reliability (ICC(3,5)>or=0.70). Although some measures had moderate reliability (ICC<0.70), they had low SEMs, which suggested high measurement precision. These findings support using these methods for examining neuromuscular activity in subjects with PFPS. PMID- 19121953 TI - When is your head at? An exploration of the factors associated with the temporal focus of the wandering mind. AB - Two experiments employed experience sampling to examine the factors associated with a prospective and retrospective focus during mind wandering. Experiment One explored the contribution of working memory and indicated that participants generally prospect when the task does not require continuous monitoring. Experiment Two demonstrated that in the context of reading, interest in what was read suppressed both past and future-related task-unrelated-thought. Moreover, in disinterested individuals the temporal focus during mind wandering depended on the amount of experience with the topic matter-less experienced individuals tended to prospect, while more experienced individuals tended to retrospect. Together these results suggest that during mind wandering participants' are inclined to prospect as long as the task does not require their undivided attention and raise the intriguing possibility that autobiographical associations with the current task environment have the potential to cue the disinterested mind. PMID- 19121954 TI - Reproducibility of imaging human knee cartilage by delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) at 1.5 Tesla. AB - OBJECTIVE: The purpose of this study was to investigate the day-to-day reproducibility of the delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) measurement at different knee joint surfaces in healthy subjects at 1.5 Tesla (T). METHODS: The dGEMRIC experiment was repeated for 10 asymptomatic volunteers three times with an average interval of 5 days between scans. The measurement was performed from a single sagittal slice through the center of the lateral femoral condyle and from the center of the patella in the axial plane. Cartilage was manually segmented into superficial, deep and full-thickness regions of interests (ROIs) at different topographical locations of the femur, tibia and patella. The reproducibility was evaluated separately for each ROI as well as for the entire bulk cartilage in the slice of each joint surface. RESULTS: The reproducibility at various ROIs expressed by root-mean-square average coefficient of variation (CV(RMS)) ranged between 4.7-12.9%. Thirty out of thirty-three ROIs showed a CV(RMS) less than 10%. Intraclass correlation coefficient (ICC) ranged between 0.45 and 0.98. The CV(RMS) and ICC for bulk dGEMRIC were 4.2% and 0.95 for femur, 5.5% and 0.87 for tibia, and 4.8% and 0.97 for patella. CONCLUSIONS: The dGEMRIC technique showed good day-to-day reproducibility, on the average 8% for small deep or superficial segments, 7% for full-thickness ROIs and 5% for bulk ROIs covering all visible cartilage in a single joint surface. We conclude that dGEMRIC imaging at field strength 1.5 T can be used as a reliable instrument for the assessment of articular cartilage when staff has been carefully trained. PMID- 19121955 TI - Horizontal alveolar ridge augmentation using distraction osteogenesis: comparison with a bone-splitting method in a dog model. AB - OBJECTIVE: Although horizontal alveolar distraction osteogenesis (DO) appears to be a viable option for implant placement in a narrow alveolar ridge, relatively few reports exist on horizontal DO. Horizontal DO using a titanium mesh plate is similar to alveolar bone splitting, but without a bone graft. Therefore, this study compared 2 methods of bone augmentation, horizontal DO and a bone-splitting (BS) method with bone graft, using an experimental dog model. STUDY DESIGN: Five beagle dogs were studied. The experiments were designed in a spit-mouth manner. One side of the alveolar ridge of the mandible was 4 mm expanded using horizontal DO, followed by implant placement 2 months after augmentation. The other side of the ridge of the mandible was 4 mm augmented using a bone splitting (BS) method with a bone graft combined with simultaneous implant placement. RESULTS: The average amount of bone gain on the DO side (2.7 mm) was significantly greater than that on the BS side (1.7 mm) (P = .008) The keratinized soft tissue gain on the DO side (2.8 mm) was significantly greater than that on the BS side (0.6 mm) (P = .02). Resonance frequency analysis revealed that implants placed in the distracted area achieved good stability compared with those on the BS side. The implants were fully embedded in mature lamellar bone, and direct bone contact with the implant surface was seen 3 months after implant placement in the distracted area. CONCLUSIONS: Horizontal alveolar ridge distraction appears be an effective technique for the placement of implants in a narrow alveolar ridge. PMID- 19121956 TI - Fixation properties of a biodegradable "free-form" osteosynthesis plate with screws with cut-off screw heads: biomechanical evaluation over 26 weeks. AB - OBJECTIVE: The aim of this study was to compare the postoperative fixation properties of a biodegradable osteosynthesis "free-form" plate achieved with countersunk screws with those provided by screws with cut-off screw heads. STUDY DESIGN: Acrylic pipes were fixed together to simulate fracture fixation for tensile testing. Additional plates were fixed to a polyurethane block with a single screw for plate-screw pullout testing. Specimens were incubated in phosphate buffer solution at 37 degrees C, and testing was conducted at various time points during hydrolytic degradation of 26 weeks. In both tests the specimens were loaded at a speed of 5 mm/min until failure. The yield load, maximum load, and stiffness were recorded, and failure mode was visually determined. RESULTS: Both countersunk screws and screws with cut-off screw heads provided similar plate fixation properties over degradation time. CONCLUSION: According to these results, fixation of the biodegradable osteosynthesis free form plate with screws with cut-off screw heads seems to be feasible. PMID- 19121957 TI - Traumatic intrusion of primary teeth and its effects on the permanent successors: a clinical follow-up study. AB - OBJECTIVE: Owing to the anatomic proximity between the root of the primary tooth and its permanent successor, trauma to primary dentition may cause developmental disturbances in unerupted permanent teeth. STUDY DESIGN: This report presents the findings of a prospective 7-year study examining developmental disturbances of permanent teeth attributed to intrusive injury of their primary predecessors based on initial patient records and clinical observation. Intrusive injuries were diagnosed and grouped according to World Health Organization classifications, as modified by Andreasen et al. RESULTS: A total of 78 children (aged 12-48 months) presenting with intrusive trauma to 138 primary incisors were available for follow-up examinations. The most frequently intruded primary teeth were maxillary incisors (93.47%), with the right central primary incisor representing 41.3% of all intruded teeth. In 23 patients, 36 teeth were scheduled for extraction during the first visit. During the course of follow-up, 41 of the remaining teeth exhibited post-traumatic consequences, including pulp necrosis (78.0%), internal and/or external root resorption (14.6%), ankylosis (4.9%), and obliteration of the pulp canal (2.5%). In 74 permanent successors (53.6%), >or=1 of the following developmental disturbances were observed: enamel hypoplasia (28.3%), crown and/or root deformation (16.7%), and ectopic eruption (16.7%). CONCLUSION: There was no significant correlation between age of intrusion and frequency of subsequent developmental disturbances (P > .05). PMID- 19121958 TI - Pediatric mucosa-associated lymphoid tissue (MALT) lymphoma of lip: a case report and literature review. AB - Mucosa-associated lymphoid tissue (MALT) lymphoma is listed in the new World Health Organization classification as extranodal marginal zone B-cell lymphoma of MALT. Most cases occur predominantly in adults and are rare in children or adolescents. We report, with a review of earlier literature, a MALT lymphoma of the lower lip in a 7-year-old immunocompetent child. The patient had a sessile, indurated, and firm mass on the lower lip without regional lymphadenopathy. Microscopically, dense lymphoid cell infiltrates composed of small- to medium sized centrocyte-like cells and plasmacytoid cells infiltrating around scattered reactive germinal centers were observed, with formation of typical lymphoepithelial lesions. Immunohistochemical staining demonstrated positivity of atypical lymphoid cells for CD20, CD43, CD79a, and Bcl-2 and negativity for CD3, CD5, CD10, CD23, Ki-1, Bcl-10, and ALK. Cytoplasmic kappa light chain restriction was demonstrated. We conclude that this case represents the youngest patient with a MALT lymphoma of the lip. PMID- 19121959 TI - Oral bacterial cultures in nontraumatic brain abscesses: results of a first-line study. AB - OBJECTIVE: Bacterial cultures from nontraumatic brain abscesses (BAs) frequently contain oral bacteria. We assessed bacterial cultures from BAs and oral infective sources for a bacterial match. STUDY DESIGN: Bacterial samples from brain abscesses and oral abscesses, and at sites with probing depths >or=3.5 mm were taken from 11 nontraumatic BA patients and analyzed. RESULTS: Brain abscess bacterial cultures were obtained in 9 of the 11 cases, which revealed 5 cases of Streptococcus milleri group bacteria and 4 cases of subgingival flora. The bacteriologic results were interpreted taking all medical and bacteriologic findings into account, which made an oral origin of the BAs most likely in 6 of the 11 cases: from an oral abscess and from the subgingival flora in 3 cases each. CONCLUSIONS: Early collaboration between neurosurgeons, infectious disease specialists, and oral-maxillofacial surgeons will aid the identification and treatment of suspected oral sources of nontraumatic BAs. PMID- 19121960 TI - Evolution of CBCT: the tree now has two distinct branches. PMID- 19121961 TI - Intraosseous cystic lesions of the jaws in children: a retrospective analysis of 47 consecutive cases. AB - OBJECTIVES: The aim was to review the characteristics and treatment approach in 47 cases of intraosseous cystic lesions of the jaws in children. STUDY DESIGN: Forty-three patients, 2-14 years old, with 47 cystic lesions of the jaws, were treated during the period 2000-2007. All cysts were surgically removed. No bone grafts were used and the teeth involved were preserved whenever possible. Removed tissues were examined histologically. Follow-up period ranged from 6 months to 7 years. RESULTS: In 74.5% of the cases, cysts were enucleated, and in 17.0% they were marsupialized. Most commonly, the cysts were dentigerous (20, 42.6%), followed by eruption cysts, odontogenic keratocysts, and radicular cysts (10.6% each) and buccal bifurcation cysts (8.6%). CONCLUSION: Cystic lesions in children were found to be mainly of developmental origin. Treatment was surgical removal without interfering, when possible, with the development of the dentition. Surgical approach was usually enucleation and, to a lesser extent, marsupialization. PMID- 19121962 TI - Positron-emission tomography imaging in the diagnosis of bisphosphonate-related osteonecrosis of the jaw. AB - OBJECTIVES: The purpose of this study was to investigate the sensitivity of F-18 fluoride and F-18 fluorodeoxyglucose (FDG) positron-emission tomography (PET) in the diagnosis of bisphosphonate-related osteonecrosis of the jaw (BRONJ), and to test their suitability for assessing the severity of BRONJ. STUDY DESIGN: Nine patients with BRONJ were studied using F-18 fluoride and F-18 FDG PET. For analysis, 8 regions of interest (ROI) were defined in the jaws of each patient. Maximum count rates for each ROI in both PET examinations were analyzed. RESULTS: In both studies, increased tracer enhancement was observed in regions with BRONJ. Uptake of fluoride significantly exceeded that of FDG. FDG uptake increased systematically, but not significantly, with increasing severity of BRONJ. CONCLUSION: F-18 fluoride PET is a sensitive method for diagnosis of BRONJ. FDG PET could be useful for evaluation of the severity of BRONJ. Further studies are required to prove the specificity of the methods. PMID- 19121963 TI - Patient discomfort and retakes in periapical examination of mandibular third molars using digital receptors and film. AB - OBJECTIVE: The aim was to compare patient discomfort and evaluate the frequency of retakes using intraoral digital receptors and conventional film for radiographic examination of mandibular third molars. STUDY DESIGN: Both mandibular third molar regions were examined in 110 patients with 2 of 5 digital intraoral receptors. Discomfort was scored on a visual analog scale (VAS) for each receptor and for film as a reference. If the whole tooth was not imaged on the digital image, a retake was performed using film. t Tests evaluated differences in VAS score, chi-squared tests evaluated differences in frequency of remakes, and logistic regression analyses evaluated factors predisposing for retake. RESULTS: No significant difference existed in VAS scores between right and left sides for film (P = .24). The digital receptors were more uncomfortable than film (P < .001), and CDR-APS was more uncomfortable than Digora (P = .049) and Vista (P = .002). The frequency of retakes was higher for solid-state sensors than photostimulable phosphor plates (PSPs) (P < .018). Gender (P = .022), type of receptor (P < .021), and VAS score (P = .001) were predisposing factors for a retake. CONCLUSION: Patients accepted film better than digital intraoral receptors, and retake frequency was lower for PSPs compared with solid-state sensors. PMID- 19121964 TI - Histologic and tomographic analyses of the temporomandibular joint after mandibular advancement surgery: study in minipigs. AB - OBJECTIVE: The aim was to elucidate the changes occurring in temporomandibular joint (TMJ) after surgical mandibular advancement with different fixation materials: bicortical screws and miniplates. STUDY DESIGN: Eighteen minipigs were randomly divided into 3 groups: group I (control), nonoperated animals; group II, animals submitted to surgical advancement surgery and osteosynthesis by bicortical screws; and group III, animals submitted to surgical advancement surgery and osteosynthesis by miniplates. Four months after the surgeries, TMJs were collected and histologically prepared after computerized tomography (CT) scanning for the blind detection of erosion, flattening, and osteophyte. RESULTS: The CT analysis revealed significant alterations in the shape of the condyles (erosion: P = .0010; flattening: P < .0001) for group II compared with groups I and III. Descriptive histologic analysis was compatible with the CT findings. CONCLUSIONS: The results indicated that bicortical screw fixation resulted in more pronounced condylar alterations in the shape of the condyles than miniplate osteosynthesis. However, further clinical studies are necessary to confirm these data. PMID- 19121965 TI - Multi-exponential signal decay from diffusion in a single compartment. AB - Multi-exponential decays in diffusion experiments are typically fitted to sums of exponentially decaying components; often this is taken as evidence for spins in multiple distinct compartments. Here we examine the signal decay due to diffusion in a single cylinder, for short diffusion times (lightly restricted). The signals are well-modeled by a sum of two exponentials, despite the single compartment housing the spins. The results agree with a previous theoretical examination of the problem. The implication for biological systems is that multiple decay signal components may not correspond to multiple physical compartments. PMID- 19121966 TI - Prevalence of low bone mass in relation to estrogen treatment and body composition in male-to-female transsexual persons. AB - Bone health is a parameter of interest in the daily follow-up of male-to-female (M --> F) transsexual persons both before and after sex reassignment surgery (SRS) due to an intensely changing hormonal milieu. We have studied body composition, areal, geometric, and volumetric bone parameters, using DXA and peripheral quantitative computed tomography at different sites in 50 M --> F transsexual persons, at least 3 yr after the start of the hormonal treatment and 1 yr after SRS. In this cross-sectional study, hormone levels and markers of bone metabolism were assessed using immunoassays. Prevalence of low bone mass as defined by a Z-score < or = -2.0 according to DXA criteria was 26% at lumbar spine and 2% at the total hip. We found no major differences in hormonal parameters between participants with a Z-score < or = or > -2.0. Markers of bone turnover were comparable between subjects with or without low bone mass, indicating a stable bone turnover at the time of investigation. No significant differences in bone size or density were observed between patients on transdermal vs. oral estrogens. Low bone mass is not uncommon in M --> F transsexual persons. Smaller bone size, and a strikingly lower muscle mass compared with men appear to underlie these findings. PMID- 19121967 TI - Transcriptomic and network component analysis of glycerol kinase in skeletal muscle using a mouse model of glycerol kinase deficiency. AB - Glycerol kinase (GK) is at the interface of fat and carbohydrate metabolism and has been linked to obesity and type 2 diabetes mellitus (T2DM). The purpose of this study was to investigate the role of GK in fat metabolism and insulin signaling in skeletal muscle (an important end organ tissue in T2DM). Microarray analysis determined that there were 525 genes that were differentially expressed (1.2-fold, p value<0.05) between knockout (KO) and wild-type (WT) mice. Quantitative PCR (qPCR) confirmed the differential expression of genes including glycerol kinase (Gyk), phosphatidylinositol 3-kinase regulatory subunit, polypeptide 1 (p85 alpha) (Pik3r1), insulin-like growth factor 1 (Igf1), and growth factor receptor bound protein 2-associated protein 1 (Gab1). Network component analysis demonstrated that transcription factor activities of myogenic differentiation 1 (MYOD), myogenic regulatory factor 5 (MYF5), myogenin (MYOG), nuclear receptor subfamily 4, group A, member 1 (NUR77) are decreased in the Gyk KO whereas the activity of paired box 3 (PAX3) is increased. The activity of MYOD was confirmed using a DNA binding assay. In addition, myoblasts from Gyk KO had less ability to differentiate into myotubes compared to WT myoblasts. These findings support our previous studies in brown adipose tissue and demonstrate that the role of Gyk in muscle is due in part to its non-metabolic (moonlighting) activities. PMID- 19121968 TI - First case of Pasteurella multocida endocarditis of the tricuspid valve: a favorable outcome following medical treatment. AB - We describe a rare case of tricuspid valve endocarditis caused by Pasteurella multocida in an elderly woman with no previous history of valvular heart disease. Risk factors included contact with animals and old age. The patient was treated successfully with five days of intravenous ampicillin followed by four weeks of oral ciprofloxacin. PMID- 19121970 TI - Macrophage subsets in mechanical brain injury (MBI)--a contribution to timing of MBI based on immunohistochemical methods: a pilot study. AB - Cortical hemorrhages as a consequence of closed mechanical brain injury (MBI) trigger an inflammatory response including a distinct increase of macrophages. According to published data this reactive macrophage population is heterogenous as to their immunological properties. The expression of certain immunohistochemically detectable epitopes of macrophages, however, may correlate with the posttraumatic interval (PTI). In a pilot study, 50 selected cases of cortical hemorrhages with 1 min to 1.5 years PTI were examined by light microscopy and macrophages were labeled with CD68-, HLA-D-, HAM-56-, LN-5-, and 25F9-antibodies, while hemosiderin was detected by a Prussian-blue reaction. Qualitative and semiquantitative investigations were performed. The semiquantitative study included 5 different classes. The results of the study revealed a distinct timetable of the appearance of macrophages labeled with certain antibodies. While HLA-D immunoreactivity was detected after a PTI of 6h in the cortex and white matter bordering the traumatic hemorrhage, CD68 immunopositive macrophages were present after 12h, LN-5 and HAM-56 after 48h, and 25F9 within 10d. Hemosiderin-containing macrophages were detectable within 100h in the same region. Within the hemorrhage itself a certain immunoreactivity of macrophages starts several hours before: CD68 after 3h, LN-5 after 24h, HAM-56 after 31h, hemosiderin after 76h, and 25F9 after 4d. For forensic purposes these observations are of crucial importance because the time course of the appearance of certain immunopositive macrophages labeled with different antibodies allows a differentiated timing of contusional injuries; however, the cause of this different immunopositive reaction remains unexplained. The observed time dependency of different macrophage antigen expressions in cortical hemorrhages after closed head injury is a suitable method to estimate the PTI and will allow a forensic reliable estimation if future investigations are extended on higher numbers of cases and/or additional markers. PMID- 19121972 TI - A facile synthesis of pyridazinone derivatives under ultrasonic irradiation. AB - A new, efficient and general method for preparation of N-substituted pyridazinones using ultrasound irradiation is reported. Under ultrasound the reaction time decreases substantially, the yields are high and the reaction conditions are mild. It was noticed that substituents at the 3-(6)-position of pyridazone heterocycle have a substantial influence on the reactivity, while the effect of the substituents at the 1-(2)-position seems to be of minor importance. A comparative study of the reactions performed under ultrasound conditions versus at room temperature has been done. PMID- 19121971 TI - Bioluminescent bacteria have potential as a marker of drowning in seawater: two immersed cadavers retrieved near estuaries. AB - We detected numerous bioluminescent bacteria in blood samples from two cadavers that had been immersed in estuarine environments. Autopsy, diatomaceous and toxicological findings indicated death by drowning, which agreed with environmental aspects and the findings of police investigations. Bioluminescent bacteria appeared in blood samples cultured on selective agar containing 2%, 3% and 4% NaCl after about 18h. Blood from the left side of the heart, the right side of the heart and the femoral vein generated 7.0 x 10(2), 2.0 x 10(4) and 8.0 x 10(2) cfu/ml of blood (case 1), and 1.8 x 10(4), 1.1 x 10(3) and 2.5 x 10(1) cfu/ml (case 2) of bioluminescent colonies, respectively, in agar containing 4% NaCl. Homologous analysis based on the 16S rRNA gene also identified the bioluminescent colonies as Vibrio fischeri and V. harveyi, which normally inhabit seawater. This simple assay might serve as an additional indicator to support a conclusion of death by drowning together with the diatom test. PMID- 19121973 TI - Density and type of food retailers surrounding Canadian schools: variations across socioeconomic status. AB - Lower socioeconomic status (SES) neighbourhoods may have differential access to food retailers, potentially explaining the varying area-level obesity rates. The food retail environment around 188 schools across Canada was examined, including full-service restaurants, fast food restaurants, sub/sandwich retailers, donut/coffee shops, convenience stores, and grocery stores. School addresses were linked to census data to obtain area-level SES measures. Access to food retailers was generally not associated with the neighbourhood SES in the immediate proximity. Within the broader neighbourhood, lower SES neighbourhoods had access to fewer food retailers of all types. This effect was diminished after taking population density into account. PMID- 19121974 TI - The association between degenerative hip joint pathology and size of the gluteus maximus and tensor fascia lata muscles. AB - The aim of this study was to obtain, using Magnetic Resonance Imaging (MRI), muscle volume measurements for the gluteus maximus (upper: UGM and lower: LGM portions) and tensor fascia lata (TFL) muscles in both healthy subjects (n=12) and those with unilateral osteoarthritis (OA) of the hip (mild: n=6, and advanced: n=6). While control group subjects were symmetrical between sides for the muscles measured, subjects with hip joint pathology showed asymmetry in GM muscle volume dependent on stage of pathology. The LGM demonstrated atrophy around the affected hip in subjects with advanced pathology (p<0.05), however asymmetry of the UGM (p<0.01) could be attributed largely to hypertrophy on the unaffected side, based on between group comparisons of muscle volume. TFL showed no significant asymmetry, or difference compared to the normal control group. This study highlights the functional separation of UGM and LGM, and the similarities of the UGM and TFL, both superficial abductors appearing to maintain their size around the affected hip. Further research is required to determine the specific changes occurring in the deeper abductor muscles. This information may assist in the development of more targeted and effective exercise programmes in the management of OA of the hip. PMID- 19121975 TI - Scaled quantum chemical calculations and FTIR, FT-Raman spectral analysis of 2 Methylpyrazine. AB - The mid and far FTIR and Raman spectra of 2-Methylpyrazine (2MP) were measured in the liquid state. The fundamental vibrational frequencies and intensity of vibrational bands were evaluated using density functional theory (DFT) and standard B3LYP/6-311+G** basis set combination. The vibrational spectra were interpreted, with the aid of normal coordinate analysis based on a scaled quantum mechanical (SQM) force field. The infrared and Raman spectra were also predicted from the calculated intensities. Comparison of simulated spectra with the experimental spectra provides important information about the ability of the computational method to describe the vibrational modes. Unambiguous vibrational assignment of all the fundamentals was made using the total energy distribution (TED). PMID- 19121976 TI - Modulation of human exteroceptive jaw reflexes during simulated mastication. AB - OBJECTIVE: To investigate changes in synaptic input from lower lip afferents to human jaw muscle motoneurons during simulated mastication. METHODS: The lower lip of 14 subjects was stimulated electrically under static and dynamic conditions. In the static condition, subjects bit at mid-open position and received stimuli while keeping the masseteric excitation level at 20%, 40%, 60%, 80%, or 100% of the maximum EMG (generated during simulated chewing). In the dynamic condition, the subjects 'masticated' at their habitual chewing pace, and stimuli were delivered whenever the jaw crossed a predetermined gape. In both conditions, mildly (scores of 2-3 on a 0-10 rating scale) and moderately (scores of 5-6) painful stimulus intensities were used. RESULTS: Under static conditions, there was no modulation of the inhibitory masseteric reflexes with the level of the background level of excitation used in these experiments. However, under dynamic conditions there were significant strength modulations with gape that differed between mildly and moderately painful stimuli. CONCLUSIONS: Reflexes in response to mildly painful stimuli were 'gated' during simulated mastication: as the teeth moved closer toward occlusion, the inhibitory response was progressively reduced. Conversely, responses to moderately painful stimuli became stronger as the teeth moved closer toward occlusion. SIGNIFICANCE: The modulation described allows smooth mastication to occur as it gates out mildly painful signals while responding strongly when the signal indicates potential or actual damage closer to occlusion. PMID- 19121977 TI - Motor imagery and action observation: modulation of sensorimotor brain rhythms during mental control of a brain-computer interface. AB - OBJECTIVE: This study investigates the impact of a continuously presented visual feedback in the form of a grasping hand on the modulation of sensorimotor EEG rhythms during online control of a brain-computer interface (BCI). METHODS: Two groups of participants were trained to use left or right hand motor imagery to control a specific output signal on a computer monitor: the experimental group controlled a moving hand performing an object-related grasp ('realistic feedback'), whereas the control group controlled a moving bar ('abstract feedback'). Continuous feedback was realized by using the outcome of a real-time classifier which was based on EEG signals recorded from left and right central sites. RESULTS: The classification results show no difference between the two feedback groups. For both groups, ERD/ERS analysis revealed a significant larger ERD during feedback presentation compared to an initial motor imagery screening session without feedback. Increased ERD during online BCI control was particularly found for the lower alpha (8-10 Hz) and for the beta bands (16-20, 20-24 Hz). CONCLUSIONS: The present study demonstrates that visual BCI feedback clearly modulates sensorimotor EEG rhythms. When the feedback provides equivalent information on both the continuous and final outcomes of mental actions, the presentation form (abstract versus realistic) does not influence the performance in a BCI, at least in initial training sessions. SIGNIFICANCE: The present results are of practical interest for classifier development and BCI use in the field of motor restoration. PMID- 19121978 TI - Yo-Yo intermittent recovery test versus the Universite de Montreal Track Test: relation with a high-intensity intermittent exercise. AB - The first purpose of this study was to determine whether the peak velocity (V(Yo Yo)) achieved during the Yo-Yo intermittent recovery test (Yo-Yo) and the maximal aerobic velocity (MAV) determined from the Universite de Montreal Track Test (UMTT) could be used interchangeably. The second purpose was to check that the V(Yo-Yo) is related to the intermittent exercise performance, which consisted of repeated 90 m distance runs in 15s performed until exhaustion, alternated with 15s of passive recovery (15/15). Fourteen amateur soccer players performed, in a random order, the 15/15 and two incremental field-tests: the Yo-Yo and the UMTT. The results of this study showed that MAV was significantly correlated to the V(Yo-Yo) (r=0.79, p<0.01). However, the error was not constant, when the V(Yo-Yo) and the MAV values were higher than 16.3 km h(-1), the MAV values tends to be higher than the V(Yo-Yo), while when the V(Yo-Yo) and the MAV values were lower than 16.3 km h(-1), the MAV values tends to be lower than the V(Yo-Yo). MAV and V(Yo-Yo) were significantly correlated to the time to exhaustion of the 15/15 (r=0.74 and r=0.72, respectively) and show that both tests are similarly related to the high-intensity intermittent exercise performance. PMID- 19121979 TI - Allogeneic stem cell transplantation for severe aplastic anemia: graft rejection remains a problem. AB - We reviewed the outcome in 15 consecutive patients with severe aplastic anemia with a median age of 23 years who received matched sibling peripheral blood stem cell transplantation. Conditioning regimen was cyclophosphamide (Cy)+anti thymocyte globulin (ATG). Cumulative incidence of transplant related mortality, graft failure, acute and chronic GVHD were 20%, 33%, 25%, and 8.3%, respectively. Conditioning with Cy only, resulted in higher rejection rate compared to Cy plus ATG (75% versus 12.5%, p=0.03). Eighty percent of patients are alive with a median follow-up of 19.5 (4.6-35.6) months. Two of the three patients who were re transplanted with fludarabine had sustained donor chimerism. PMID- 19121980 TI - Determination of human urinary organophosphate flame retardant metabolites by solid-phase extraction and gas chromatography-tandem mass spectrometry. AB - Organophosphorus flame retardants (OPFR), phosphorus triesters, are widely used chemicals with a high share of the worldwide flame retardant market. In animal experiments, dialkyl- and diarylphosphates are the main metabolites of OPFR. Therefore we elaborated a GC-MS/MS-method for the detection of OPFR-metabolites in human urine after solid phase extraction and derivatization with pentafluorobenzylbromide. The limits of detection range from 0.1 to 1 microg/l. Interday imprecision were 2-8%. The applicability of the method is shown by determination of the internal burden of 30 persons of the German general population. OPFR-metabolite concentrations range from asbestos (ASB)>concrete (RCC) for the roof structures. The estimated radon levels in the study area are relatively higher than the country's average value of 23 Bq m(-3) and global average value of 30 Bq m(-3). PMID- 19122000 TI - Growth hormone inhibition of glucose uptake in adipocytes occurs without affecting GLUT4 translocation through an insulin receptor substrate-2 phosphatidylinositol 3-kinase-dependent pathway. AB - Growth hormone (GH) pretreatment of 3T3-L1 adipocytes resulted in a concentration and time-dependent inhibition of insulin-stimulated glucose uptake. Surprisingly, this occurred without significant effect on insulin-stimulated glucose transporter (GLUT) 4 translocation or fusion with the plasma membrane. In parallel, the inhibitory actions of chronic GH pretreatment also impaired insulin dependent activation of phosphatidylinositol (PI) 3-kinase bound to insulin receptor substrate (IRS)-2 but not to IRS-1. In addition, insulin-stimulated Akt phosphorylation was inhibited by GH pretreatment. In contrast, overexpression of IRS-2 or expression of a constitutively active Akt mutant prevented the GH induced insulin resistance of glucose uptake. Moreover, small interfering RNA mediated IRS-2 knockdown also inhibited insulin-stimulated Akt activation and glucose uptake without affecting GLUT4 translocation and plasma membrane fusion. Together, these data support a model in which chronic GH stimulation inhibits insulin-dependent activation of phosphatidylinositol 3-kinase through a specific interaction of phosphatidylinositol 3-kinase bound to IRS-2. This inhibition leads to suppression of Akt activation coupled to glucose transport activity but not translocation or plasma membrane fusion of GLUT4. PMID- 19122002 TI - Radiation doses in paediatric interventional cardiology procedures. AB - The objective was to investigate paediatric doses in coronary angiography (CA) and percutaneous transluminal coronary angioplasty (PTCA) in the largest cardiac hospital in Greece. Forty procedures were carried out by two board-certified senior interventional cardiologists. Data collected were: patient weight, height, age, fluoroscopy time (FT), total number of images (N) and kerma-area product (KAP). Median (range) age was 7.5 y (17 d to 17 y). Median FT, N and KAP were 4 min, 655, 2.1 Gy cm2 for CA and 12.1 min, 1296, 14.7 Gy cm2 for PTCA (corresponding adult diagnostic reference levels (DRLs) are: 6.5 min, 700, 45 Gy cm2 for CA and 15.5 min, 1000 and 85 Gy cm2 for PTCA). The highest percentage of cine dose was in newborns (0-1 y) (CA: 92% and PTCA: 100%). As age increased, cine dose percentage decreased, whereas total radiation dose increased. Median paediatric FT and N recorded reached or even exceeded adult DRL and should be optimised. Paediatric DRL should be set. PMID- 19122003 TI - Conformity versus Innovation. PMID- 19122004 TI - Activation of (Na+ + K+)-ATPase modulates cardiac L-type Ca2+ channel function. AB - Cellular Ca(2+) signaling underlies diverse vital biological processes, including muscle contractility, memory encoding, fertilization, cell survival, and cell death. Despite extensive studies, the fundamental control mechanisms that regulate intracellular Ca(2+) movement remain enigmatic. We have found recently that activation of the (Na(+)+K(+))-ATPase markedly potentiates intracellular Ca(2+) transients and contractility of rat heart cells. Little is known about the pathway responsible for the activation of the (Na(+)+K(+))-ATPase-initiated Ca(2+) signaling. Here, we demonstrate a novel mechanism in which activation of the (Na(+)+K(+))-ATPase is coupled to increased L-type Ca(2+) channel function through a signaling cascade involving Src and ERK1/2 but not well established regulators of the channel, such as adrenergic receptor system or activation of PKA or CaMKII. We have also identified Ser(1928), a phosphorylation site for the alpha1 subunit of the L-type Ca(2+) channel that may participate in the activation of the (Na(+)+K(+))-ATPase-mediated Ca(2+) signaling. The findings reported here uncover a novel molecular cross-talk between activation of the (Na(+)+K(+))-ATPase and L-type Ca(2+) channel and provide new insights into Ca(2+) signaling mechanisms for deeper understanding of the nature of cellular Ca(2+) handling in heart. PMID- 19122005 TI - Engineering a stable and selective peptide blocker of the Kv1.3 channel in T lymphocytes. AB - Kv1.3 potassium channels maintain the membrane potential of effector memory (T(EM)) T cells that are important mediators of multiple sclerosis, type 1 diabetes mellitus, and rheumatoid arthritis. The polypeptide ShK-170 (ShK-L5), containing an N-terminal phosphotyrosine extension of the Stichodactyla helianthus ShK toxin, is a potent and selective blocker of these channels. However, a stability study of ShK-170 showed minor pH-related hydrolysis and oxidation byproducts that were exacerbated by increasing temperatures. We therefore engineered a series of analogs to minimize the formation of these byproducts. The analog with the greatest stability, ShK-192, contains a nonhydrolyzable phosphotyrosine surrogate, a methionine isostere, and a C terminal amide. ShK-192 shows the same overall fold as ShK, and there is no evidence of any interaction between the N-terminal adduct and the rest of the peptide. The docking configuration of ShK-192 in Kv1.3 shows the N-terminal para phosphonophenylalanine group lying at the junction of two channel monomers to form a salt bridge with Lys(411) of the channel. ShK-192 blocks Kv1.3 with an IC(50) of 140 pM and exhibits greater than 100-fold selectivity over closely related channels. After a single subcutaneous injection of 100 microg/kg, approximately 100 to 200 pM concentrations of active peptide is detectable in the blood of Lewis rats 24, 48, and 72 h after the injection. ShK-192 effectively inhibits the proliferation of T(EM) cells and suppresses delayed type hypersensitivity when administered at 10 or 100 microg/kg by subcutaneous injection once daily. ShK-192 has potential as a therapeutic for autoimmune diseases mediated by T(EM) cells. PMID- 19122006 TI - Selective serotonin reuptake inhibitor use and risk of gestational hypertension. AB - OBJECTIVE: The purpose of this study was to assess the effects of treatment with selective serotonin reuptake inhibitors (SSRIs) on the risks of gestational hypertension and preeclampsia. METHOD: The authors analyzed data from 5,731 women with nonmalformed infants and no underlying hypertension who participated in the Slone Epidemiology Center Birth Defects Study from 1998 to 2007. Gestational hypertension was defined as incident hypertension diagnosed after 20 weeks of pregnancy, with and without proteinuria (i.e., with and without preeclampsia). The risks of gestational hypertension and preeclampsia were compared between women who did and did not receive SSRI treatment during pregnancy. Relative risks and 95% confidence intervals (CIs) were estimated using the Cox proportional hazards model, adjusting for prepregnancy sociodemographic, lifestyle, reproductive, and medical factors. RESULTS: Gestational hypertension was present in 9.0% of the 5,532 women who were not treated with SSRIs and 19.1% of the 199 women who were treated with SSRIs. Among women who received treatment, gestational hypertension was present in 13.1% of the 107 women who received treatment only during the first trimester and in 26.1% of the 92 women who continued treatment beyond the first trimester. The occurrence of preeclampsia was 2.4% among women who were not treated with SSRIs, 3.7% among women who were exposed to SSRIs only during the first trimester, and 15.2% among women who continued SSRI treatment beyond the first trimester. Relative to women who did not receive treatment, the adjusted relative risk of preeclampsia was 1.4 for women who discontinued treatment and 4.9 for women who continued treatment. CONCLUSION: SSRI exposure during late pregnancy-whether a causal factor or not might identify women who are at an increased risk for gestational hypertension and preeclampsia. Further investigation is needed in order to separate the effects of treatment with SSRIs from those of underlying mood disorders. PMID- 19122007 TI - Anticipatory activation in the amygdala and anterior cingulate in generalized anxiety disorder and prediction of treatment response. AB - OBJECTIVE: The anticipation of adverse outcomes, or worry, is a cardinal symptom of generalized anxiety disorder. Prior work with healthy subjects has shown that anticipating aversive events recruits a network of brain regions, including the amygdala and anterior cingulate cortex. This study tested whether patients with generalized anxiety disorder have alterations in anticipatory amygdala function and whether anticipatory activity in the anterior cingulate cortex predicts treatment response. METHOD: Functional magnetic resonance imaging (fMRI) was employed with 14 generalized anxiety disorder patients and 12 healthy comparison subjects matched for age, sex, and education. The event-related fMRI paradigm was composed of one warning cue that preceded aversive pictures and a second cue that preceded neutral pictures. Following the fMRI session, patients received 8 weeks of treatment with extended-release venlafaxine. RESULTS: Patients with generalized anxiety disorder showed greater anticipatory activity than healthy comparison subjects in the bilateral dorsal amygdala preceding both aversive and neutral pictures. Building on prior reports of pretreatment anterior cingulate cortex activity predicting treatment response, anticipatory activity in that area was associated with clinical outcome 8 weeks later following treatment with venlafaxine. Higher levels of pretreatment anterior cingulate cortex activity in anticipation of both aversive and neutral pictures were associated with greater reductions in anxiety and worry symptoms. CONCLUSIONS: These findings of heightened and indiscriminate amygdala responses to anticipatory signals in generalized anxiety disorder and of anterior cingulate cortex associations with treatment response provide neurobiological support for the role of anticipatory processes in the pathophysiology of generalized anxiety disorder. PMID- 19122008 TI - Manic symptoms during depressive episodes in 1,380 patients with bipolar disorder: findings from the STEP-BD. AB - OBJECTIVE: Little is known about how often bipolar depressive episodes are accompanied by subsyndromal manic symptoms in bipolar I and II disorders. The authors sought to determine the frequency and clinical correlates of manic symptoms during episodes of bipolar depression. METHOD: From among 4,107 enrollees in the National Institute of Mental Health's Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD), 1,380 individuals met criteria for bipolar I or II depressive syndromes at the time of enrollment and were assessed for concomitant manic symptoms. Illness characteristics were compared in patients with pure bipolar depressed episodes and those with mixed depressive presentations. RESULTS: Two-thirds of the subjects with bipolar depressed episodes had concomitant manic symptoms, most often distractibility, flight of ideas or racing thoughts, and psychomotor agitation. Patients with any mixed features were significantly more likely than those with pure bipolar depressed episodes to have early age at illness onset, rapid cycling in the past year, bipolar I subtype, history of suicide attempts, and more days in the preceding year with irritability or mood elevation. CONCLUSIONS: Manic symptoms often accompany bipolar depressive episodes but may easily be overlooked when they appear less prominent than depressive features. Subsyndromal manic symptoms during bipolar I or II depression demarcate a more common, severe, and psychopathologically complex clinical state than pure bipolar depression and merit recognition as a distinct nosologic entity. PMID- 19122009 TI - Understanding placebo response in pediatric depression trials. PMID- 19122010 TI - Child psychiatry growin' up. PMID- 19122011 TI - Reversing criminalization. PMID- 19122012 TI - Issues for DSM-V: adding problem codes to facilitate assessment of quality of care. PMID- 19122013 TI - Activity-dependent transcription and disorders of human cognition. PMID- 19122014 TI - Treating the childhood bipolar controversy: a tale of two children. PMID- 19122015 TI - Severe eating disorder in a 28-year-old man with William's syndrome. PMID- 19122017 TI - Antipsychotic properties of muscarinic drugs. PMID- 19122018 TI - Reduction in muscarinic M1-mediated hypercholinergic state and beneficial cognitive effects of muscarinic agonists in schizophrenia. PMID- 19122019 TI - Risperidone-associated increase in triglyceride levels. PMID- 19122020 TI - Extreme elevation of creatinine phosphokinase levels in neuroleptic malignant syndrome associated with atypical antipsychotics. PMID- 19122021 TI - An early report on the role of nitrates in erectile dysfunction. PMID- 19122022 TI - Education research: Assessment of neurology resident clinical competencies in the neurology clinic. AB - BACKGROUND: Objective evaluation of neurology resident clinical skills is required by the American Board of Psychiatry and Neurology and is important to insure improvement in clinical competency throughout their residency. METHODS: In this study, neurology residents from all 3 years of training and neurology faculty independently completed a form on new clinic patients documenting their decisions on anatomic localization, diagnosis, diagnostic tests, and management. RESULTS: Compared to the attending patient evaluation, we found significant improvement in identical scoring by year of residency training. All resident years outperformed medical students in the neurology clerkship. CONCLUSION: Our clinical assessment form adds one more tool to the list of currently used assessment methods to evaluate resident clinical competency. PMID- 19122023 TI - Teaching NeuroImage: Inverted V sign in subacute combined degeneration of spinal cord. PMID- 19122025 TI - Levels of evidence: Taking Neurology to the next level. PMID- 19122026 TI - The need for biomarkers in amyotrophic lateral sclerosis drug development. PMID- 19122027 TI - A novel ALS2 splice-site mutation in a Cypriot juvenile-onset primary lateral sclerosis family. AB - BACKGROUND: Primary lateral sclerosis (PLS) is a rare neurodegenerative disease that affects the upper motor neurons of the CNS. Juvenile-onset PLS (JPLS) is inherited in an autosomal recessive mode and is also found in sporadic cases. A consanguineous Cypriot family with three affected individuals presenting with JPLS was identified and studied. METHODS: Patients were clinically evaluated and samples were taken from consenting family members. All available family members were genotyped and linkage analysis at marker loci spanning the wider region of the ALS2 gene was performed. Selected exons of the ALS2 gene were sequenced and RNA analysis was performed using available lymphoblastoid cell lines from the proband. RESULTS: All affected individuals presented in the second year of life with progressive upper motor neuron dysfunction, affecting both bulbar and extremity muscles. Severity was variable, with two of the patients remaining ambulatory in the second and fifth decade of life while the third one was never able to walk. A novel ALS2 homozygous c.2980-2A>G mutation at the splice acceptor site of intron 17 was identified and its effect was confirmed at the RNA level. CONCLUSIONS: This novel ALS2 splice-site mutation is causing the loss of exon 18 in the transcript which results in a frameshift after exon 17. This frameshift most probably introduces a stop codon seven amino acids further down the new reading frame (p.993fsX7) and is expected to lead to a premature stop in exon 19 thus leading to a truncated protein after translation. PMID- 19122028 TI - Long-term trends in carpal tunnel syndrome. AB - OBJECTIVE: To assess temporal trends in carpal tunnel syndrome (CTS) incidence, surgical treatment, and work-related lost time. METHODS: Incident CTS and first time carpal tunnel release among Olmsted County, Minnesota, residents were identified using the medical records linkage system of the Rochester Epidemiology Project; 80% of a sample were confirmed by medical record review. Work-related CTS was identified from the Minnesota Department of Labor and Industry. RESULTS: Altogether, 10,069 Olmsted County residents were initially diagnosed with CTS in 1981-2005. Overall incidence (adjusted to the 2000 US population) was 491 and 258 per 100,000 person-years for women vs men (p < 0.0001) and 376 per 100,000 for both sexes combined. Adjusted annual rates increased from 258 per 100,000 in 1981 1985 to 424 in 2000-2005 (p < 0.0001). The average annual incidence of carpal tunnel release surgery was 109 per 100,000, while that for work-related CTS was 11 per 100,000. An increase in young, working-age individuals seeking medical attention for symptoms of less severe CTS in the early to mid-1980s was followed in the 1990s by an increasing incidence in elderly people. CONCLUSIONS: The incidence of medically diagnosed carpal tunnel syndrome (CTS) accelerated in the 1980s. The cause of the increase is unclear, but it corresponds to an epidemic of CTS cases resulting in lost work days that began in the mid-1980s and lasted through the mid-1990s. The elderly present with more severe disease and are more likely to have carpal tunnel surgery, which may have significant health policy implications given the aging population. PMID- 19122029 TI - Frontal FDG-PET activity correlates with cognitive outcome after STN-DBS in Parkinson disease. AB - BACKGROUND: Inconsistent changes of cognitive functioning have been reported in patients with Parkinson disease (PD) with deep brain stimulation (DBS) of the subthalamic nucleus (STN). To investigate the underlying pathomechanisms, we correlated alterations of cognitive test performance and changes of neuronal energy metabolism in frontal basal ganglia projection areas under bilateral STN stimulation. METHODS: We conducted verbal fluency, learning, and memory tests and 18-fluorodeoxyglucose (FDG) PET in nine patients with PD with STN-DBS before and 6 months after surgery. Using coregistered MRI, postoperative changes of the normalized cerebral metabolic rates of glucose (nCMRGlc) in the dorsolateral prefrontal cortex (DLPFC), lateral orbitofrontal cortex (LOFC), ventral and dorsal cingulum (v/dACC), and in Broca area were determined and correlated with alterations of neuropsychological test results. RESULTS: After surgery, highly variable changes of both cognitive test performance and frontal nCMRGlc values were found with significant correlations between verbal fluency and FDG uptake in the left DLPFC (Brodmann area [BA] 9, 46), left Broca area (BA 44/45), and the right dACC (BA 32). A decrease of nCMRGlc in the left OFC (BA 11/47) and dACC (BA 32) correlated with a decline of verbal learning. All patients showed reduced metabolic activity in the right anterior cingulate cortex after DBS. Baseline cognitive abilities did not predict verbal learning or fluency changes after surgery. CONCLUSIONS: These data show a significant linear relationship between changes in frontal 18-fluorodeoxyglucose PET activity and changes in cognitive outcome after deep brain stimulation of the subthalamic nucleus (STN) in advanced Parkinson disease. The best correlations were found in the left frontal lobe (dorsolateral prefrontal cortex and Broca area). Baseline performance on cognitive tests did not predict cognitive or metabolic changes after STN electrode implantation. PMID- 19122030 TI - Longitudinal prognostic value of serum "free" copper in patients with Alzheimer disease. AB - BACKGROUND: Serum copper not bound to ceruloplasmin ("free") appears slightly elevated in patients with Alzheimer disease (AD). We explored whether a deregulation of the free copper pool can predict AD clinical worsening. METHODS: We assessed levels of copper, iron, zinc, transferrin, ceruloplasmin, peroxides, total antioxidant capacity, free copper, and apolipoprotein E genotype in 81 patients with mild or moderate AD, mean age 74.4, SD = 7.4 years, clinically followed up after 1 year. The association among biologic variables under study and Mini-Mental State Examination (MMSE) (primary outcome), activities of daily living (ADL), and instrumental activities of daily living (IADL) (secondary outcomes) performed at study entry and after 1 year were analyzed by multiple regression. RESULTS: Free copper predicted the annual change in MMSE, adjusted for the baseline MMSE by means of a linear regression model: it raised the explained variance from 2.4% (with only sex, age, and education) to 8.5% (p = 0.026). When the annual change in MMSE was divided into < 3 or > or = 3 points, free copper was the only predictor of a more severe decline (predicted probability of MMSE worsening 23%: odds ratio = 1.23; 95% confidence interval = 1.03-1.47; p = 0.022). Hyperlipidemic patients with higher levels of free copper seemed more prone to worse cognitive impairment. Free copper at baseline correlated with the ADL and IADL clinical scales scores at 1 year. CONCLUSIONS: These results show an association between copper deregulation and unfavorable evolution of cognitive function in Alzheimer disease. Further research is needed to establish whether copper is an independent risk factor for cognitive decline. PMID- 19122031 TI - Microglial activation and amyloid deposition in mild cognitive impairment: a PET study. AB - BACKGROUND: Activated microglia may play a role in the pathogenesis of Alzheimer disease (AD) as they cluster around beta-amyloid (Abeta) plaques. They are, therefore, a potential therapeutic target in both AD and its prodrome amnestic mild cognitive impairment (MCI). OBJECTIVE: To characterize in vivo with (11)C (R)-PK11195 and (11)C-PIB PET the distribution of microglial activation and amyloid deposition in patients with amnestic MCI. METHODS: Fourteen subjects with MCI had (11)C-(R)-PK11195 and (11)C-PIB PET with psychometric tests. RESULTS: Seven out of 14 (50%) patients with MCI had increased cortical (11)C-PIB retention (p < 0.001) while 5 out of 13 (38%) subjects with MCI showed increased (11)C-(R)-PK11195 uptake. The MCI subgroup with increased (11)C-PIB retention also showed increased cortical (11)C-(R)-PK11195 binding (p < 0.036) though this increase only remained significant in frontal cortex after a correction for multiple comparisons. There was no correlation between regional levels of (11)C (R)-PK11195 and (11)C-PIB binding in individual patients with MCI: only three of the five MCI cases with increased (11)C-(R)-PK11195 binding had increased levels of (11)C-PIB retention. CONCLUSIONS: Our findings indicate that, while amyloid deposition and microglial activation can be detected in vivo in around 50% of patients with mild cognitive impairment (MCI), these pathologies can occur independently. The detection of microglial activation in patients with MCI suggests that anti-inflammatory therapies may be relevant to the prevention of AD. PMID- 19122032 TI - Diagnostic utility of different MRI and MR angiography measures in Fabry disease. AB - BACKGROUND: Neurologic hallmarks of Fabry disease (FD) include small fiber neuropathy as well as cerebral micro- and macroangiopathy with premature stroke. Cranial MRI shows progressive white matter lesions (WML) at an early age, increased signal intensity in the pulvinar, and tortuosity and dilatation of the larger vessels. To unravel the most promising imaging tool for the detection of CNS involvement in FD we compared the diagnostic utility of the different MR imaging findings. METHODS: Twenty-five clinically affected patients with FD (age 36.5 +/- 11.0) and 20 age-matched controls were investigated by structural MRI, MR angiography, and diffusion tensor imaging (DTI). Individual WML volumes, global mean diffusivity (MD), and mean cerebral artery diameters were determined. RESULTS: Using receiver operating characteristic analyses, enlarged diameters of the following cerebral arteries significantly separated patients with FD from controls: middle cerebral artery: area under curve (AUC) = 0.75, p = 0.005; posterior cerebral artery: AUC = 0.69, p = 0.041; carotid artery: 0.69, p = 0.041; basilar artery: AUC = 0.96, p < 0.0005. A total of 87% of the individuals were correctly classified by basilar artery diameters (sensitivity 95%, specificity 83%). WML volumes and global MD values did not significantly separate patients from controls. CONCLUSIONS: With an accuracy of 87%, basilar artery diameters were superior to all other MR measures for separating patients with Fabry disease (FD) from controls. Future studies should adopt basilar artery measurements for early detection and monitoring of brain involvement in FD. Moreover, further investigations should reveal if the dilated vasculopathy in FD could be a screening marker to detect FD in a cohort of other cerebrovascular diseases, especially in cryptogenic stroke. PMID- 19122033 TI - Smoking and family history and risk of aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Smoking and family history of aneurysmal subarachnoid hemorrhage (aSAH) are independent risk factors for aSAH. Using a population-based case control study of hemorrhagic stroke, we hypothesized that having both a first degree relative with a brain aneurysm or SAH (+FH) and current smoking interact to increase the risk of aSAH. METHODS: Cases of aneurysmal SAH were prospectively recruited from all 17 hospitals in the five-county region around the University of Cincinnati. Controls were identified by random digit dialing. Controls were matched to cases of aSAH by age (+/-5 years), race, and sex. Conditional multiple logistic regression was used to identify independent risk factors. For deviation from the additive model, the interaction constant ratio test was used. RESULTS: A total of 339 cases of aSAH were matched to 1,016 controls. Compared to current nonsmokers with no first-degree relatives with aSAH (-FH), the odds ratio (OR) for aSAH for current nonsmokers with +FH was 2.5 (95% confidence interval [CI] 0.9-6.9); for current smokers with -FH, OR = 3.1 (95% CI 2.2-4.4); and for current smokers with +FH, OR = 6.4 (95% CI 3.1-13. 2). The interaction constant ratio, which measured the deviation from the additive model, was significant: 2.19 (95% CI 0.80-5.99). The lower bound of the 95% CI >0.5 signifies a departure from the additive model. CONCLUSION: Evidence of a gene-environment interaction with smoking exists for aneurysmal subarachnoid hemorrhage. This finding is important to counseling family members and for screening of intracranial aneurysm (IA) as well as the design and interpretation of genetic epidemiology of IA studies. PMID- 19122035 TI - Delusional misidentifications and duplications: right brain lesions, left brain delusions. AB - When the delusional misidentification syndromes reduplicative paramnesia and Capgras syndromes result from neurologic disease, lesions are usually bifrontal and/or right hemispheric. The related disorders of confabulation and anosognosis share overlapping mechanisms and anatomic pathology. A dual mechanism is postulated for the delusional misidentification syndromes: negative effects from right hemisphere and frontal lobe dysfunction as well as positive effects from release (i.e., overactivity) of preserved left hemisphere areas. Negative effects of right hemisphere injury impair self-monitoring, ego boundaries, and attaching emotional valence and familiarity to stimuli. The unchecked left hemisphere unleashes a creative narrator from the monitoring of self, memory, and reality by the frontal and right hemisphere areas, leading to excessive and false explanations. Further, the left hemisphere's cognitive style of categorization, often into dual categories, leads it to invent a duplicate or impostor to resolve conflicting information. Delusions result from right hemisphere lesions. But it is the left hemisphere that is deluded. PMID- 19122036 TI - Georges Marinesco and the early research in neuropathology. AB - OBJECTIVE: To present the scientific contributions of Georges Marinesco (1863 1938) and place his achievements within the context of early neuropathology research. BACKGROUND: Neuropathology is a relatively recent medical field, its origins dating to the late 19th century. RESULTS: One of the most important neuroscientists of that period was the Romanian-born Georges Marinesco. He became a neurologist under Charcot's guidance at the Salpetriere Hospital, in Paris. In 1892, Paul Blocq and Marinesco gave a first account of senile plaques, having used their pathologic skills in the examination of nine deceased epileptic patients. They did not, however, relate the plaques to dementia. Marinesco made discoveries in neuropathology which he described from a histopathologic perspective, and introduced new medical terms such as neuronophagia, chromatolysis, and medullomyoblastoma. He also drew correlations between clinical neurologic findings and morphology, for example in congenital cerebellar ataxia, syringomyelia, and parkinsonism. From 1899 he used cinematography as a medical research tool. CONCLUSION: Marinesco was a prolific researcher in the field of neuropathology, especially neurodegeneration but also in clinical neurology. He is now considered the founder of the modern Romanian school of neurology. PMID- 19122034 TI - Oral fingolimod (FTY720) in multiple sclerosis: two-year results of a phase II extension study. AB - OBJECTIVE: To report the results of a 24-month extension of a phase II trial assessing the efficacy, safety, and tolerability of the once-daily oral sphingosine-1-phosphate receptor modulator, fingolimod (FTY720), in relapsing multiple sclerosis (MS). METHODS: In the randomized, double-blind, placebo controlled core study, 281 patients received placebo or FTY720, 1.25 or 5.0 mg/day, for 6 months. During the subsequent dose-blinded extension, patients assigned to placebo were re-randomized to either dose of FTY720; those originally assigned to FTY720 continued at the same dose. Patients receiving FTY720 5.0 mg were switched to 1.25 mg during the month 15 to month 24 study visits. RESULTS: Of 281 patients randomized in the core study, 250 (89%) entered the extension phase, and 189 (75.6%) received treatment for 24 months. During the core study, FTY720 significantly reduced gadolinium-enhanced (Gd(+)) lesions and annualized relapse rate (ARR) compared with placebo, with no differences between doses. During the extension phase, patients who switched from placebo to FTY720 showed clear reductions in ARR and lesion counts compared with the placebo phase; ARR and lesion counts remained low in patients who continued FTY720 treatment. After 24 months, 79 to 91% of patients were free from Gd(+) lesions and up to 77% of patients remained relapse free. FTY720 was well tolerated; no new safety concerns emerged during months 7 to 24 compared with the 6-month core study. CONCLUSIONS: Once-daily oral treatment with FTY720, 1.25 or 5.0 mg, for up to 2 years, was well tolerated and was associated with low relapse rates and lesion activity. PMID- 19122037 TI - Postpartum thrombosis of a developmental venous anomaly. PMID- 19122038 TI - A new centronuclear myopathy phenotype due to a novel dynamin 2 mutation. PMID- 19122039 TI - Efalizumab-induced isolated cerebral lupus-like syndrome. PMID- 19122040 TI - T-cell neurolymphomatosis involving cauda equina and sciatic nerves. PMID- 19122042 TI - Multiple sclerosis and cannabis: a cognitive and psychiatric study. PMID- 19122041 TI - Clinical spectrum of voltage-gated potassium channel autoimmunity. PMID- 19122043 TI - The Ninth International Workshop on the Biology of Lactation in Farm Animals and Triennial Lactation Symposium. PMID- 19122044 TI - Genetic diversity and pedigree analysis of the Finnsheep breed. AB - Genetic diversity in the Finnsheep breed was analyzed by quantifying the demographic trends, the depth of known pedigree, effective population size, and the amount of inbreeding, as well as identifying candidate rams within the current population for future breeding and conservation purposes. Pedigree records of 148,833 animals with a pedigree completeness coefficient > or =0.60 and born from 1989 to 2006 were used to estimate the parameters. Mean inbreeding coefficient increased by 0.10% (P < 0.001) and 0.15% (P < 0.001) per annum in all animals and breeding (i.e., reproducing) animals, respectively. Average relationship coefficients among rams, among ewes, and between rams and ewes in breeding animals increased over time and reached 1.67, 1.45, and 1.46% in the 2005 cohort, respectively. The average for breeding rams was above the other 2 averages in almost all birth years. The observed generally low average relationship coefficients between rams and ewes indicate that no extra restrictions on the use of the breeding animals are needed in the near future. Average generation interval was 2.85 yr in the studied period, and the effective population size was estimated to be 119 and 122 using different methods. Relationship coefficients of rams with other breeding rams and rams with breeding ewes are suggested to aid in situ and ex situ conservation decisions on maintaining genetic diversity of Finnsheep. PMID- 19122045 TI - Preweaning performance and body composition of calves from straightbred Nellore and Bos taurus x Nellore crosses. AB - The objectives were to evaluate preweaning performance, body composition, and efficiency of calves representing straightbred Nellore (NL), F(1), and 3-breed cross systems. Energy requirements, milk production, and efficiency of 39 cow calf pairs were recorded from straightbred NL calves from NL cows (10), crossbred (Angus-sired) calves from NL cows (ANL: 9), and crossbred calves (CC; Canchim sired: 5/8 Charolais, 3/8 Zebu) from ANL (10) and Simmental x NL (10) cows. Cows and their respective calves were individually fed from birth to weaning (17 to 190 d postpartum). At 38 d of age, corn silage (7.8% CP, 2.19 Mcal of ME/kg of DM) was available to calves ad libitum. Milk production at 42, 98, 126, and 180 d postpartum was recorded by weighing calves before and after suckling. The ratio between GE and ME of milk was considered 1:0.93. Calves were slaughtered at weaning and the 9th-, 10th-, and 11th-rib section was removed for body composition estimation. The ANL calves were lighter (P < 0.01) at birth than the CC calves; the NL calves were intermediate. At weaning, the CC calves were heavier (P = 0.04) than the NL and ANL calves (230 +/- 5.5 vs. 172 +/- 8.1 and 209 +/- 8.6 kg, respectively). The ANL calves had greater (371 +/- 27 Mcal; P = 0.01) silage intake than the NL (270 +/- 25 Mcal) and CC (279 +/- 17 Mcal) calves. Milk energy intake was greater for the CC calves (970 +/- 38 Mcal of ME; P = 0.005) than the NL (670 +/- 57 Mcal of ME) and ANL (743 +/- 61 Mcal of ME) calves. The ANL calves compensated for the reduced milk production of the NL cows, which supplied less of their energy requirement for growth by increased silage intake. Calves from crossbred cows received a greater proportion of their total energy intake from milk. Crossbred calves had greater (P < 0.03) retained energy (retained energy = weaning body energy - birth body energy) than the NL calves (388 +/- 23 for ANL, and 438 +/- 15 for CC vs. 312 +/- 22 Mcal for NL calves). Percentages of water (P = 0.74) and chemical fat (P = 0.51) were similar among groups (63.7 +/- 0.6 and 14.3 +/- 0.7% for ANL calves, 63.1 +/- 0.4 and 14.7 +/- 0.5% for CC calves, and 63.3 +/- 0.6 and 13.7 +/- 0.7% of empty BW for water and chemical fat, respectively, for NL calves). Energetic efficiency (kcal of retained energy/Mcal of ME intake) was similar (P = 0.52) among groups (358 +/ 22 for ANL calves, 355 +/- 14 for CC calves, and 327 +/- 22 for NL calves). The greater BW gains and the differences in empty body composition at weaning were not enough to compensate for the greater ME intake of crossbreds. In this study, the crossbreeding systems evaluated increased preweaning calf performance but did not affect gross or energetic calf efficiency. PMID- 19122046 TI - Variability in the behavior of kids born of primiparous goats during the first hour after parturition: effect of the type of parturition, sex, duration of birth, and maternal behavior. AB - The aim of the study was to determine the effect of the type of birth, the sex of the kids, the duration of the birth (categorized as short, medium, or long), and the level of maternal care (categorized as low, medium, or high) on the behavioral variables of kids during the first hour after birth. The parturitions of 78 primiparous goats of Murciano-Granadina breed (46 single-birth and 32 twin birth) along with the behavior of the kids (44 males and 66 females) during the first hour of life were studied. Birth weight and duration of parturition were greater in single-birth kids (2.94 kg and 60.5 min, respectively) than in twin birth kids (2.27 kg and 43.2 min, respectively). Birth weight and duration of parturition was greater in males (2.74 kg and 54.61 min) than in females (2.43 kg and 47.70 min). All the kids attempted to stand during the first hour of life, but only 83% attempted to suckle with 65% succeeding. Single-birth kids attempted to stand earlier than twin-birth kids (7.05 vs. 9.08 min), although they achieved this later (16.87 vs. 13.21 min). Compared with twin-birth kids, single-birth kids attempted to suckle later (22.45 vs. 34.76 min, respectively) and achieved it later (25.69 vs. 37.32 min). In the single-birth kids the duration of the first suckling was shorter (16.11 vs. 22.26 s), although total suckling time was greater (5.86 min) than in the twin-birth kids. Males tried to stand sooner than females (7.41 vs. 8.78 min), but took longer (16.12 vs. 13.81 min). The sex factor had no significant effect on suckling-related variables. Compared with medium- and long-duration-birth kids, short-duration-birth kids attempted to suckle earlier, (29.34, 34.23, and 12.82 min, respectively), achieved suckling earlier (31.75, 37.00, and 16.70 min, respectively), and suckled longer at first attempt (0.32, 0.17, and 0.45 min, respectively). Total suckling time was longer in long-duration-birth kids than in medium- and short-duration birth (9.07, 2.63, and 3.86 min, respectively). These results suggest that heavier kids suffer more during birth and are slower to achieve suckling; however, they appear to recover quickly because the total suckling time observed during the first hour was greater. PMID- 19122047 TI - Dynamics of offensive gas-phase odorants in fresh and aged feces throughout the development of beef cattle. AB - Livestock odors are largely caused by several groups of volatile organic compounds (VOC), including sulfur-containing compounds, VFA, and phenols and indoles. Throughout the growth stages of cattle in the nursery and feedlot, distinctly different diets are formulated to meet the changing requirements of the animal. Because diet composition and manure management are 2 major factors affecting odor emissions, it was assumed that changes in diet composition along the development of calves would affect VOC emissions from fresh and stored manure. In this study, the dynamics of gas-phase VOC in feces from 6 Holstein Friesian bull calves were followed at 5 ages: 1 to 5 wk (stage I), 6 to 8 wk (stage II, before weaning), 9 to 14 wk (stage III, after weaning), and 15 to 36 wk (stages IV and V). The CP content of the formulated diet decreased from 23.0 to 13.9%. Samples of fresh feces were incubated under either aerobic or anaerobic conditions for 21 d. The VOC were analyzed from the feces headspace by solid phase microextraction, followed by gas chromatography-mass spectrometry. Distinct changes in gas-phase VOC were observed in fresh and aged feces from calves at different ages. Semiquantitatively (based on comparative peak area counts), the following trends were observed: 1) S-containing compounds were the least dominant in fresh feces at the age of 2 wk (stage I), whereas VFA esters were the most dominant. 2) At the age of 7 wk (stage II), 1 wk before calves were weaned, feces seemed to be the most offensive, presumably because of the difficulty of synchronizing the requirements of the animal and the diet formulation during this stage of rapid development. 3) The VOC decreased during storage of feces under aerobic conditions but significantly increased at all 5 life stages during storage under anaerobic conditions. This study demonstrates that life stage and manure management affect odor emissions from beef fattening operations. Incorporation of the age and diet of calves in odor modeling could improve annoyance predictions. PMID- 19122048 TI - The propositional approach to associative learning as an alternative for association formation models. AB - Associative learning effects can be defined as changes in behavior that are due to relations between events in the world. Most often, these effects are explained in terms of the formation of unqualified associations in memory. I describe an alternative theoretical explanation, according to which associative learning effects are the result of the nonautomatic generation and evaluation of propositions about relations between events. This idea is supported by many studies showing that associative learning effects are determined not only by the direct experience of events but also by prior knowledge, instructions, intervention, and deductive reasoning. Moreover, evidence supports the assumption that associative learning effects depend on nonautomatic processes. Whereas a propositional approach thus offers many new insights, questions can be raised about what the idea of association formation adds to our understanding of associative learning. PMID- 19122049 TI - What are association formation models? AB - In his presentation of the propositional account of associative learning, De Houwer (2009) argues that association formation models (AFMs) assume excitatory link representations and automatic learning processes. However, the application of AFMs to human causal and contingency learning has assumed propositional forms of representation, although excitatory link representations are also required to explain certain nonrational consequences of associative learning. Moreover, at least two of the AFMs that have been applied to human associative learning invoke processing with nonautomatic characteristics. In conclusion, the distinction between the propositional account and AFMs of associative learning lies not in the form of representations but in the specific details of the learning processes generating the associative representations. PMID- 19122051 TI - Attentional changes in blocking are not a consequence of lateral inhibition. AB - In three human causal learning experiments, we examined attentional modulation in the blocking task, in which participants typically learn little about a novel cue B when it is paired with a previously trained, predictive cue A. Evidence indicates that this blocking training led to a decrement in attention to the blocked cue B. The present experiments addressed whether this decrease in attention to the blocked cue could be better explained as being due to lateral inhibition from the pretrained cue A to the blocked cue B, or as a cue-specific property that is not conditional on the presence or absence of other stimuli. Strong effects of learned predictiveness were observed on participants' causal judgments (Experiment 1) and choice behavior (Experiments 2 and 3). However, no evidence for lateral inhibitory processes emerged in any of the experiments, despite explicit attempts to maximize experimental sensitivity to this effect. The results are discussed in the context of formal models of the operation of attentional processes in human and animal learning. PMID- 19122052 TI - Factors moderating blocking in human place learning: the role of task instructions. AB - Cognitive map theory assumes that novel environmental information is automatically incorporated into existing cognitive maps as a function of exploration. Reports of blocking in place learning cast doubt on this claim. In these studies, subjects were first trained to find a place, using a set of landmarks (Set A). Then novel landmarks (Set B) were added for additional trials. Subsequent removal of the Set A landmarks showed that the novel landmarks alone were insufficient for successful navigation. We investigated whether instructing human subjects to explore the environment can moderate blocking. First, we demonstrated that blocking is absent in a computer implementation of the Morris water maze (MWM) in which subjects are instructed to explore. We then studied why others found blocking in a different MWM implementation, in which the task instructions did not suggest exploration. In experiments that faithfully replicated this MWM variant, we found that subjects did not acquire cognitive maps and that blocking was attenuated when instructions were provided that encouraged exploration. Together, these findings indicate that blocking in human place learning may reflect a performance deficit, not a learning deficit, and that instructions can moderate blocking. Our results thus support the automatic update assumption of cognitive map theory. PMID- 19122054 TI - Pigeons' memory for time: assessment of the role of subjective shortening in the duration-comparison procedure. AB - Pigeons were trained in a duration-comparison procedure to peck one key if the comparison duration (c) was 1 sec shorter than a standard duration (s), and another key if c was 1 sec longer than s. During training, the s-c delay was 1 sec, and the total duration of an s-c pair was not predictive of the correct choice. In Experiment 1, during equal-duration pair test trials, pigeons increasingly responded long (i.e., c > s) as the s-c delay was lengthened. In Experiment 2, we demonstrated that s affected long responding on equal-duration test trials, even at the 8-sec s-c delay. In Experiment 3, long responding increased as the s-c delay was lengthened, even when stimulus conditions during the s-c delay differed from those during the intertrial interval (ITI). Additional analyses indicated that it was unlikely that the increase in long responding was due to the pigeons' adding the s-c delay to c and comparing the total against the duration of s. The increase in long responding with an increase in s-c delay is more consistent with subjective shortening of s than with confusion between the s-c delay and the ITI. PMID- 19122053 TI - Spacing extinction trials alleviates renewal and spontaneous recovery. AB - Studies of extinction in classical conditioning situations can reveal techniques that maximize the effectiveness of exposure-based behavior therapies. In three experiments, we investigated the effect of varying the intertrial interval during an extinction treatment in a fear-conditioning preparation with rats as subjects. In Experiment 1, we found less fear at test (i.e., more effective extinction) when extinction trials were widely spaced, relative to intermediate or massed extinction trials. In Experiment 2, we used an ABA renewal procedure and observed that spaced trials attenuated renewal of conditioned fear relative to massed trials. In Experiment 3, we used a similar design, but instead of changing the physical context at the time of testing, we interposed a retention interval after the extinction treatment to produce a change in the temporal context. The results showed less spontaneous recovery of fear after spaced than after massed extinction trials. These results suggest that extinction is more enduring when the extinction trials are spaced rather than massed. Although the benefits of spacing trials are small when there is no contextual change from extinction to testing, a change in either physical or temporal context following massed extinction trials leads to a recovery from extinction, which is reduced when the trials are spaced. PMID- 19122055 TI - Extinction of conditioned inhibition: effects of different outcome continua. AB - For some decades, failures to find extinction of inhibition through unpaired presentations of the inhibitor were taken as evidence against conceptualizing inhibition as the symmetrical counterpart of excitation. Recently, however, our group has demonstrated successful extinction of inhibition in human causal learning. In two experiments, we replicated and strengthened this finding by using an outcome continuum that could take on negative, neutral, or positive values. In contrast, the use of a dichotomous outcome continuum (either neutral or positive) resulted in the well-known nonoccurrence of extinction. Extinction of inhibition through the pairing of inhibitors with neutral outcomes was assessed by (1) comparing the (presumably) extinguished inhibitor with a second inhibitor that had not been presented with a neutral outcome in the extinction stage, and (2) demonstrating the course of extinction in participants' predictions. PMID- 19122056 TI - Contextual control of first- and second-learned excitation and inhibition in equally ambiguous stimuli. AB - In two three-phase experiments, rats received a final third excitatory (Experiment 1) or inhibitory (Experiment 2) phase of conditioning with a tone. The third phase came immediately prior to a test with the tone, either in the context where the tone was trained or in a different context. Groups differed in each experiment with respect to the first two phases. Rats in Groups EIE (Experiment 1) and EII (Experiment 2) received excitatory conditioning with the tone in Phase 1, followed by inhibitory conditioning with the tone. Rats in Groups IEE (Experiment 1) and IEI (Experiment 2) received inhibitory conditioning in Phase 1, followed by excitatory conditioning in Phase 2. Thus, the association being expressed in Phase 3 was consistent either with what was learned first about the stimuli or with what was learned second. Contrary to expectations, the association being expressed at the end of Phase 3, either excitatory or inhibitory, was affected by a context change, regardless of its consistency with what was learned first about the CS. PMID- 19122057 TI - Symmetrical generalization decrements: configural stimulus processing in human contingency learning. AB - Models of associative learning differ in their predictions concerning the symmetry of generalization decrements. Whereas Pearce's (1994) configural model predicts the same response decrement after adding elements to and after removing elements from a previously trained stimulus, elemental models, such as the replaced elements model and Harris's (2006) model, anticipate more of a decrement for removing than for adding elements. In three contingency learning experiments, we manipulated the motion and the spatial arrangement of colored dots in order to induce configural or elemental processing by perceptual grouping. The results reliably showed symmetrical decrements for the added and removed groups. The manipulations of the stimuli had no effect on stimulus processing. This is in line with Pearce's configural model, but it is at variance with the elemental models and previous studies. PMID- 19122058 TI - Evaluations of labrador retrievers with exercise-induced collapse, including response to a standardized strenuous exercise protocol. AB - Clinical and metabolic variables were evaluated in 14 Labrador retrievers with exercise-induced collapse (EIC) before, during, and following completion of a standardized strenuous exercise protocol. Findings were compared with previously reported variables from 14 normal Labrador retrievers that participated in the same protocol. Ten of 14 dogs with EIC developed an abnormal gait during evaluation, and these dogs were significantly more tachycardic and had a more severe respiratory alkalosis after exercise compared to the normal dogs. Muscle biopsy characteristics and sequential lactate and pyruvate concentrations were normal. Genetic testing and linkage analysis excluded malignant hyperthermia as the cause of EIC. Common causes of exercise intolerance were eliminated, but the cause of collapse in EIC was not determined. PMID- 19122059 TI - Adjuvant CCNU (lomustine) and prednisone chemotherapy for dogs with incompletely excised grade 2 mast cell tumors. AB - The use of adjuvant 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU; lomustine) to treat incompletely excised canine mast cell tumors (MCTs) has not been evaluated. Medical records of 12 dogs with grade 2 MCT treated with incomplete surgical excision and adjuvant CCNU and prednisone chemotherapy were reviewed. Local recurrence rate, metastasis rate, and survival time were evaluated. None of the dogs developed local recurrence or regional/ distant metastases. Two dogs developed fatal liver failure. The 1- and 2-year progression free rates of surviving dogs were 100% and 77%, respectively. Postoperative adjuvant CCNU appears to be a useful alternative to radiation therapy for incompletely excised canine cutaneous MCTs. PMID- 19122060 TI - Effects of carprofen and morphine on the minimum alveolar concentration of isoflurane in dogs. AB - The minimum alveolar concentration (MAC) of isoflurane in dogs was determined following carprofen (2.2 mg/kg per os) alone, morphine (1 mg/kg intravenously) alone, carprofen and morphine, and no drug control in eight healthy adult dogs. Isoflurane MAC following administration of morphine alone (0.81%+/-0.18%) or carprofen and morphine (0.68%+/-0.31%) was significantly less than the control MAC (1.24%+/-0.15%). Isoflurane MAC after carprofen alone (1.13%+/-0.13%) was not significantly different from the control value. Results indicated that administration of morphine alone or in combination with carprofen significantly reduced the MAC of isoflurane in dogs. The isoflurane MAC reduction was additive between the effects of carprofen and morphine. PMID- 19122061 TI - An expedited palliative radiation protocol for lytic or proliferative lesions of appendicular bone in dogs. AB - Fifty-eight dogs with lytic or proliferative bone lesions were treated with a radiation protocol of two 8-Gy fractions over 2 consecutive days. The protocol was well tolerated, with no increase in early or late effects over previously published protocols. Forty-three (91%) of 47 dogs responded positively to radiation, with a median time of 2 days to onset of pain relief. Median duration of pain relief was 67 days (range 12 to 503 days; mean 99+/-16 days). Median survival time for all dogs was 136 days (mean 179+/-18 days). Distal radial location was a positive prognostic indicator for survival (P=0.005). PMID- 19122062 TI - Use of single-agent carboplatin as adjuvant or neoadjuvant therapy in conjunction with amputation for appendicular osteosarcoma in dogs. AB - Survival following amputation and administration of single-agent carboplatin for treatment of appendicular osteosarcoma (OSA) in dogs was retrospectively examined. Records of 155 dogs with appendicular OSA treated with amputation and single-agent carboplatin were included from 14 centers. Any carboplatin dosage, number of doses, and protocol schedule were eligible for inclusion. The median disease-free interval (DFI) was 256 days. The median overall survival time was 307 days. Similar prognostic survival factors were identified in this study as reported in prior studies of canine appendicular OSA. Median DFI and survival were comparable to those reported in the original Bergman et al publication. Carboplatin treatment improves the survival probability in dogs with appendicular OSA compared to amputation alone and remains an acceptable alternative to adjuvant treatment with cisplatin. PMID- 19122063 TI - Ectopic hepatic parenchyma attached to the diaphragm: simulating a pulmonary mass in a cat. AB - A case of an ectopic lobe of the liver connected to a normal diaphragm is described. A 9-year-old, castrated male cat underwent thoracotomy for a pulmonary mass. The removed mass was attached to the diaphragm that histologically was ectopic liver. The ectopic liver had no connection with the main liver. Because the occurrence of ectopic supradiaphragmatic hepatic tissue is a possibility, this should be considered as a differential diagnosis for caudal pulmonary or caudal mediastinal masses in a cat. This report describes, to the authors' knowledge, the first case of ectopic hepatic tissue attached to the diaphragm of a cat. The authors also characterize the asymptomatic clinical presentation and radiographic findings of this cat and suggest further imaging with computed tomography in unusual case presentations. PMID- 19122064 TI - Embryonal rhabdomyosarcoma in a young Maine coon cat. AB - Embryonal rhabdomyosarcomas are uncommon tumors in all domestic species, especially cats. A 14-month-old Maine coon was diagnosed with an embryonal rhabdomyosarcoma in the rectus abdominus muscle, which was treated with complete surgical excision. Although no clinical progression was noted after surgery, the cat succumbed to pulmonary metastasis within 7 months. The histological diagnosis was embryonal rhabdomyosarcoma (myotubular subtype). This category of striated muscle tumors is thought to have a more aggressive clinical course. The rapid demise of this cat even with no clinical or histological evidence of metastasis at the time of resection may indicate that, as in human medicine, adjuvant chemotherapy should be considered in conjunction with early surgical excision in preventing disease progression. PMID- 19122065 TI - Head and neck swelling due to a circumferential cicatricial scar in a dog. AB - A 4-year-old, castrated male, mixed-breed dog was evaluated because of progressive head swelling, exercise intolerance, and increasing respiratory effort of 1 month's duration. Physical examination and radiographs revealed severe edema of the head and face that was cranial to a circumferential, midcervical constriction caused by scarring related to previous removal of a foreign body. Surgical en bloc resection of the cicatricial tissue was performed, and clinical signs resolved completely after 2 months. Histopathology showed ongoing inflammation and hairs within a fibrous band. This case emphasizes that incomplete wound debridement may lead to excessive fibrous tissue proliferation and that thorough wound examination and debridement should be performed after removing circumferential cervical foreign bodies to ensure complete healing. PMID- 19122066 TI - Surgical removal of a Cuterebra larva from the cervical trachea of a cat. AB - A case of cervical intratracheal cuterebriasis is reported. The cat was presented with intermittent dyspnea of 3 days' duration. The larva was located during tracheoscopy but was not retrievable. Surgical exploration of the cervical region was performed, and the larva was removed. All clinical signs resolved upon recovery from surgery. The larva was identified as a second instar Cuterebra sp. PMID- 19122072 TI - Publication of clinical trials. PMID- 19122073 TI - Delayed internal fixation of femoral shaft fracture reduces mortality among patients with multisystem trauma. AB - BACKGROUND: Fractures of the femoral shaft are common and have potentially serious consequences in patients with multiple injuries. The appropriate timing of fracture repair is controversial. The purpose of the present study was to assess the effect of timing of internal fixation on mortality in patients with multisystem trauma. METHODS: We performed a retrospective cohort study with use of data from public and private trauma centers throughout the United States that were reported to the National Trauma Data Bank (version 5.0 for 2000 through 2004). The study included 3069 patients with multisystem trauma (Injury Severity Score, > or =15) who underwent internal fixation of a femoral shaft fracture. The time to treatment was defined in categories as the time from admission to internal fixation: t(0) (twelve hours or less), t(1) (more than twelve hours to twenty-four hours), t(2) (more than twenty-four hours to forty-eight hours), t(3) (more than forty-eight hours to 120 hours), and t(4) (more than 120 hours). The relative risk of in-hospital mortality when the four later periods were compared with the earliest one was estimated with inverse probability of treatment weighted analysis. Subgroups with serious head or neck, chest, abdominal, and additional extremity injury were investigated. RESULTS: When compared with that during the first twelve hours after admission, the estimated mortality risk was significantly lower in three time categories: t(1) (relative risk, 0.45; 95% confidence interval, 0.15 to 0.98; p = 0.03), t(3) (relative risk, 0.58; 95% confidence interval, 0.28 to 0.93; p = 0.03), and t(4) (relative risk, 0.43; 95% confidence interval, 0.10 to 0.94; p = 0.03). Patients with serious abdominal trauma (Abbreviated Injury Score, > or =3) experienced the greatest benefit from a delay of internal fixation beyond twelve hours (relative risk, 0.82 [95% confidence interval, 0.54 to 1.35] for patients with an Abbreviated Injury Score of <3, compared with 0.36 [95% confidence interval, 0.13 to 0.87] for those with an Abbreviated Injury Score of > or =3) (p value for effect modification, 0.09). CONCLUSIONS: Delayed repair of femoral shaft fracture beyond twelve hours in patients with multisystem trauma, which may allow time for appropriate resuscitation, reduces mortality by approximately 50%. Patients with serious abdominal injury benefit most from delayed treatment. These results support delaying definitive treatment of long-bone injuries in patients with multisystem trauma as a means of so-called damage-control in order to reduce adverse outcomes. PMID- 19122074 TI - Computer-assisted surgical navigation does not improve the alignment and orientation of the components in total knee arthroplasty. AB - BACKGROUND: Whether total knee arthroplasty with use of computer-assisted surgical navigation can improve the limb and component alignment is a matter of debate. We hypothesized that total knee arthroplasty with use of computer assisted surgical navigation is superior to conventional total knee arthroplasty with regard to the precision of implant positioning. METHODS: Sequential simultaneous bilateral total knee arthroplasties were carried out in 160 patients (320 knees). One knee was replaced with use of a computer-assisted surgical navigation system, and the other was replaced conventionally without use of computer-assisted surgical navigation. The two methods were compared for accuracy of orientation and alignment of the components as determined by radiographs and computed tomography scans. The mean duration of follow-up was 3.4 years. RESULTS: The mean preoperative Knee Society score was 26 points, with an improvement to 92 points postoperatively, in the computer-assisted total knee arthroplasty group and 25 points, with an improvement to 93 points postoperatively, in the conventional total knee arthroplasty group. Preoperative and postoperative ranges of motion of the knees were similar in both groups. The operating and tourniquet times were significantly longer in the computer-assisted total knee arthroplasty group than in the conventional total knee arthroplasty group (p < 0.001). The groups were not significantly different with regard to the accuracy of component positioning and the number of outliers for the various radiographic parameters (p > 0.05). CONCLUSIONS: Our data demonstrate that total knee arthroplasty with use of computer-assisted surgical navigation did not result in more accurate implant positioning than that achieved in conventional total knee arthroplasty, as determined by both radiographs and computed tomography scans. PMID- 19122075 TI - Thoracolumbar burst fractures treated with posterior decompression and pedicle screw instrumentation supplemented with balloon-assisted vertebroplasty and calcium phosphate reconstruction. AB - BACKGROUND: The treatment of unstable thoracolumbar burst fractures with short segment posterior spinal instrumentation without anterior column reconstruction is associated with a high rate of screw breakage and progressive loss of reduction. The purpose of the present study was to evaluate the functional, neurologic, and radiographic results following transpedicular, balloon-assisted fracture reduction with anterior column reconstruction with use of calcium phosphate bone cement combined with short-segment posterior instrumentation and a laminectomy. METHODS: A consecutive series of thirty-eight patients with an unstable thoracolumbar burst fracture with or without neurologic deficit were managed with transpedicular, balloon-assisted fracture reduction, calcium phosphate bone cement reconstruction, and short-segment spinal instrumentation from 2002 to 2005. Twenty-eight of the thirty-eight patients were followed for a minimum of two years. Demographic data, neurologic function, segmental kyphosis, the fracture severity score, canal compromise, the Short Form-36 score, the Oswestry Disability Index score, and treatment-related complications were evaluated prospectively. RESULTS: All thirteen patients with incomplete neurologic deficits had improvement by at least one Frankel grade. The mean kyphotic angulation improved from 17 degrees preoperatively to 7 degrees at the time of the latest follow-up, and the loss of vertebral body height improved from a mean of 42% preoperatively to 14% at the time of the latest follow-up. Screw breakage occurred in two patients, and pseudarthrosis occurred in one patient. CONCLUSIONS: The present study demonstrates that excellent reduction of unstable thoracolumbar burst fractures with and without associated neurologic deficits can be maintained with use of short-segment instrumentation and a transpedicular balloon-assisted reduction combined with anterior column reconstruction with calcium phosphate bone cement performed through a single posterior incision. The resultant circumferential stabilization combined with a decompressive laminectomy led to maintained or improved neurologic function in all patients with neurologic deficits, with a low rate of instrumentation failure and loss of correction. PMID- 19122076 TI - Continuous lumbar plexus block for postoperative pain control after total hip arthroplasty. A randomized controlled trial. AB - BACKGROUND: Continuous femoral or lumbar plexus blocks have been demonstrated to provide effective postoperative analgesia of the lower extremity following total joint arthroplasty. The purpose of this study was to compare these two techniques when used with intravenous patient-controlled analgesia and the use of patient controlled analgesia alone for postoperative pain management following unilateral primary hip arthroplasty. METHODS: Two hundred and twenty-five patients undergoing unilateral total hip arthroplasty for a diagnosis of osteoarthritis were randomly allocated into one of three postoperative treatment groups: continuous lumbar plexus block with patient-controlled analgesia, continuous femoral block with patient-controlled analgesia, and patient-controlled analgesia alone. Scores on a visual analog pain scale administered during physiotherapy twenty-four hours postoperatively were used as the primary outcome measured. Secondary outcomes included scores on a visual analog pain scale at rest, hydromorphone consumption, opioid-related side effects, complications, sensory and motor blockade, and patient satisfaction. RESULTS: Continuous lumbar plexus block significantly reduced pain scores during physiotherapy on postoperative day 1 (p < 0.0001) and day 2 (p < 0.0001) compared with either continuous femoral block or patient-controlled analgesia alone. There were no significant differences for pain at rest between the two regional analgesic techniques. Both regional anesthesia techniques significantly reduced total hydromorphone consumption (p < 0.05) and delirium (disorientation to time and/or place) compared with patient-controlled analgesia alone (p < 0.023). In addition, the use of continuous lumbar plexus block was associated with fewer patients with opioid-related side effects (p < 0.05), greater distances walked (p < 0.05), and enhanced patient satisfaction (p < 0.05) compared with the use of a continuous femoral nerve block with patient-controlled analgesia or with patient-controlled analgesia alone. CONCLUSIONS: Continuous lumbar plexus and femoral blocks significantly reduce the need for opioids and decrease related side effects. Continuous lumbar plexus block is a more effective analgesic modality than is a continuous femoral block or patient-controlled intravenous administration of hydromorphone alone during physical therapy following primary unilateral total hip arthroplasty. PMID- 19122077 TI - Risk factors for infection after knee arthroplasty. A register-based analysis of 43,149 cases. AB - BACKGROUND: Clinical studies have revealed a number of important risk factors for postoperative infection following total knee arthroplasty. Because of the small numbers of cases in those studies, there is a risk of obtaining false-negative results in statistical analyses. The purpose of the present study was to determine the risk factors for infection following primary and revision knee replacement in a large register-based series. METHODS: A total of 43,149 primary and revision knee arthroplasties, registered in the Finnish Arthroplasty Register, were followed for a median of three years. The Finnish Arthroplasty Register and the Finnish Hospital Discharge Register were searched for surgical interventions that were performed for the treatment of deep postoperative infections. Cox regression analysis with any reoperation performed for the treatment of infection as the end point was performed to determine the risk factors for this adverse outcome. RESULTS: Three hundred and eighty-seven reoperations were performed because of infection. Both partial and complete revision total knee arthroplasty increased the risk of infection as compared with the risk following primary knee replacement. Male patients, patients with seropositive rheumatoid arthritis or with a previous fracture around the knee, and patients with constrained and hinged prostheses had increased rates of infection after primary arthroplasty. Wound-related complications increased the risk of deep infection. The rate of septic failure was lower after unicondylar than after total condylar primary knee arthroplasty, but the difference was not significant. The combination of parenteral antibiotic prophylaxis and prosthetic fixation with antibiotic-impregnated cement protected against septic failure, especially after revision knee arthroplasty. Following revision total knee arthroplasty, diagnosis and prosthesis type had no effect, but previous revision for the treatment of infection and wound-healing problems predisposed to repeat revision for the treatment of infection. CONCLUSIONS: There was an increased risk of deep postoperative infection in male patients and in patients with rheumatoid arthritis or a fracture around the knee as the underlying diagnosis for knee replacement. The results of the present study suggest that the infection rate is similar after partial revision and complete revision total knee arthroplasties. Combining intravenous antibiotic prophylaxis with antibiotic-impregnated cement seems advisable in revision arthroplasty. PMID- 19122078 TI - Surgical treatment of early wound complications following primary total knee arthroplasty. AB - BACKGROUND: Wound-healing problems are a known complication after primary total knee arthroplasty. However, little is known about the clinical outcomes for patients who require surgical treatment of these early wound-healing problems. The purpose of the present study was to determine the incidence, risk factors, and long-term sequelae of early wound complications requiring surgical treatment. METHODS: The total joint registry at our institution was reviewed for the period from 1981 to 2004. All knees with early wound complications necessitating surgical treatment within thirty days after the index total knee arthroplasty were identified. The cumulative probabilities for the later development of deep infection and major subsequent surgery were determined. A case-control study in which these patients were matched with an equal number of controls was performed to attempt to identify risk factors for the development of early superficial wound complications requiring surgical intervention. RESULTS: From 1981 to 2004, 17,784 primary total knee arthroplasties were performed at our institution. Fifty nine knees were identified as having early wound complications necessitating surgical treatment within thirty days after the index arthroplasty, for a rate of return to surgery of 0.33%. For knees with early surgical treatment of wound complications, the two-year cumulative probabilities of major subsequent surgery (component resection, muscle flap coverage, or amputation) and deep infection were 5.3% and 6.0%, respectively. In contrast, for knees without early surgical intervention for the treatment of wound complications, the two-year cumulative probabilities were 0.6% and 0.8%, respectively (p < 0.001 for both comparisons). A history of diabetes mellitus was identified as being significantly associated with the development of early wound complications requiring surgical intervention. CONCLUSIONS: Patients requiring early surgical treatment for wound healing problems after primary total knee arthroplasty are at significantly increased risk for further complications, including deep infection and/or major subsequent surgery, specifically, resection arthroplasty, amputation, or muscle flap coverage. These results emphasize the importance of obtaining primary wound healing after total knee arthroplasty. PMID- 19122079 TI - Operative management of distal radial fractures with 2.4-millimeter locking plates. A multicenter prospective case series. AB - BACKGROUND: In the past decade, there has been a trend toward open reduction and internal fixation of unstable distal radial fractures. There are now more than thirty different implant designs specific for the fixation of distal radial fractures. A multicenter prospective study of a case series was conducted to determine the efficacy of the operative management of distal radial fractures stabilized with 2.4-mm locking plates. METHODS: One hundred and fifty patients were entered into the study over a two-year period. The mean age of these patients was fifty-one years, and the cohort included eighty-eight women and sixty-two men. Eighty-six patients sustained a low-energy injury. Seventy-one percent of the fractures in the series were intra-articular and were Type C according to the Muller-AO Comprehensive Classification. The follow-up evaluations, which were conducted at six weeks, six months, one year, and two years, included assessments of pain, motion, grip strength, and standard radiographs. Gartland and Werley scores were recorded at six months and one and two years, and Disabilities of the Arm, Shoulder and Hand (DASH) scores were recorded at one and two years. RESULTS: One hundred and twenty-five patients (83%) had a complete follow-up at six months; 121 (81%), at one year; and 117 (78%), at two years. Significant improvements in motion, grip strength, and patient satisfaction were observed between six months and one year, but further improvements were not seen at the two-year follow-up examination. The mean DASH score changed from a preinjury baseline of 2 points to 8 points at one year and 7 points at two years (p < 0.0001). The mean Gartland and Werley score improved significantly from 4 points at six months to 2 points at two years. Of the 102 intra-articular fractures examined in the immediate postoperative period, twenty had a step-off of < or =2 mm and seven had a step-off of >2 mm. Of the seventy one intra-articular fractures seen at two years, sixty-one had no step-off, five had a step-off of < or =2 mm, and five had a step-off of >2 mm. Twenty-seven percent of the patients showed an increase of at least one grade in radiographic signs of arthritis at two years. There were twenty-eight complications, twenty of which were considered minor. Tendon inflammation occurred in nine patients. There were two tendon ruptures, one of which was due to a prominent dorsal screw tip placed through a volar plate and one of which was due to a prominent volar plate. Loss of reduction occurred in two patients, and screw loosening occurred in two patients. CONCLUSION: Internal fixation of displaced distal radial fractures with implants featuring locking screw fixation can result in good-to-excellent outcomes with a limited number of complications. PMID- 19122080 TI - Detection of traumatic arthrotomy of the knee using the saline solution load test. AB - BACKGROUND: The saline solution load test helps to determine if a wound extends into the knee joint. Little is known about the volume of injected intra-articular saline solution that is needed to effectively rule in or rule out a traumatic arthrotomy of the knee. The purpose of the present study was to determine the appropriate volume and needle location for the diagnosis of a traumatic knee arthrotomy and to assess the effect of associated variables, including knee circumference, body mass index, and sex. METHODS: Fifty-six consecutive patients scheduled for knee arthroscopy were enrolled. A standard inferolateral arthroscopic portal was made with a single stab incision with use of a number-11 blade. Injection sites were randomized to either a superomedial or inferomedial location. The injection of normal saline solution at a rate of 5 mL/sec through an 18-gauge needle was continued while the knee was moved through a range of motion until fluid extravasated from the iatrogenic laceration. The volume of injected fluid was recorded. RESULTS: The study group included thirty-one female patients and twenty-five male patients with a combined average age of fifty years and an average body mass index of 30.9. In order to effectively diagnose 50% of the arthrotomies, 75 mL of injected fluid was needed; the volumes that were needed in order to effectively diagnose 75%, 90%, 95%, and 99% of the arthrotomies were 110, 145, 155, and 175 mL, respectively. The mean volumes of injected fluid needed for a positive result at the inferomedial and superomedial needle locations were 64.0 and 95.2 mL, respectively; this difference was significant (p = 0.01). There was no correlation between necessary injection volume and sex, body mass index, or knee circumference. CONCLUSIONS: In order to detect 95% of 1-cm inferolateral arthrotomies of the knee with use of the saline solution load test, 155 mL must be injected. An inferomedial injection location requires significantly less fluid than a superomedial injection location does for the diagnosis of inferolateral arthrotomies of the knee. PMID- 19122081 TI - Incidence of and risk factors for complications associated with halo-vest immobilization: a prospective, descriptive cohort study of 239 patients. AB - BACKGROUND: Since high rates of serious complications, such as death and pneumonia, during halo-vest immobilization have been reported, there has been a tendency of restraint with regard to the use of the halo vest. However, the rate of complications in a high-volume center with sufficient experience is unknown. Our objective was to determine the incidence of and risk factors associated with complications during halo-vest immobilization. METHODS: During a five-year period, a prospective cohort study was performed in a single, level-I trauma center that was also a tertiary referral center for spinal disorders. Data from all patients undergoing halo-vest immobilization were collected prospectively, and every complication was recorded. The primary outcome was the presence or absence of complications. Univariate regression analysis and regression modeling were used to analyze the results. RESULTS: In 239 patients treated with halo-vest immobilization, twenty-six major, seventy-two intermediate, and 121 minor complications were observed. Fourteen patients (6%) died during the treatment, although only one death was related directly to the immobilization and three were possibly related directly to the immobilization. Twelve patients (5%) acquired pneumonia during halo-vest immobilization. Patients older than sixty-five years did not have an increased risk of pneumonia (p = 0.543) or halo vest-related mortality (p = 0.467). Halo vest-related complications ranged from three patients (1%) with incorrect initial placement of the halo vest to twenty-nine patients (12%) with a pin-site infection. Pin-site infection was significantly related to pin penetration through the outer table of the skull (odds ratio, 4.34; 95% confidence interval, 1.22 to 15.51; p = 0.024). In 164 trauma patients treated only with halo-vest immobilization, cervical fractures with facet joint involvement or dislocations were significantly related to radiographic loss of alignment during follow-up (odds ratio, 2.81; 95% confidence interval, 1.06 to 7.44; p = 0.031). CONCLUSIONS: There are relatively low rates of mortality and pneumonia during halo-vest immobilization, and elderly patients do not have an increased risk of pneumonia or death related to halo-vest immobilization. Nevertheless, the total number of minor complications is substantial. This study confirms that awareness of and responsiveness to minor complications can prevent subsequent development of serious morbidities and perhaps reduce mortality. PMID- 19122082 TI - Midtarsal arthrodesis in the treatment of Charcot midfoot arthropathy. AB - BACKGROUND: Fracture-dislocation of the midfoot with collapse of the longitudinal arch is common in patients with neuropathic arthropathy of the foot. In this study, we describe a technique of midfoot arthrodesis with use of intramedullary axial screw fixation and review the results and complications following use of this technique. METHODS: A retrospective study of twenty-two patients who had undergone surgical reconstruction and arthrodesis to treat Charcot midfoot deformity was performed. Bone resection and/or osteotomy were required to reduce deformity. Axially placed intramedullary screws, inserted either antegrade or retrograde across the arthrodesis sites, were used to restore the longitudinal arch. Radiographic measurements were recorded preoperatively, immediately postoperatively, and at the time of the last follow-up and were analyzed in order to assess the amount and maintenance of correction. RESULTS: Patients were evaluated clinically and radiographically at an average of fifty-two months. Complete osseous union was achieved in sixteen of the twenty-two patients, at an average of 5.8 months. There were five partial unions in which a single joint did not unite in an otherwise stable foot. There was one nonunion, with recurrence of deformity. All patients returned to an independent functional ambulatory status within 9.5 months. Weight-bearing radiographs showed the talar-first metatarsal angle, the talar declination angle, and the calcaneal-fifth metatarsal angle to have improved significantly and to have been corrected to nearly normal values by the surgery. All measurements remained significantly improved, as compared with the preoperative values, at the time of final follow-up. There were no recurrent dislocations. Three patients had a recurrent plantar ulcer at the metatarsophalangeal joint that required additional surgery. There were eight cases of hardware failure. CONCLUSIONS: Open reduction and arthrodesis with use of multiple axially placed intramedullary screws for the surgical correction of neuropathic midfoot collapse provides a reliable stable construct to achieve and maintain correction of the deformity. PMID- 19122083 TI - Comparison of irrigation solutions and devices in a contaminated musculoskeletal wound survival model. AB - BACKGROUND: There is much to learn about the effectiveness of different methods currently used for the irrigation of open wounds. The purpose of this study was to compare various approaches in a survival animal model. METHODS: We used an established goat model involving the creation of a reproducible complex musculoskeletal wound followed by inoculation with Pseudomonas aeruginosa (lux) bacteria. This genetically altered luminescent bacterium provides the ability for quantitative analysis with a photon-counting camera system. For Study 1, wound irrigation was performed six hours after the injury and inoculation; the goats were assigned to four treatment groups: normal saline solution, bacitracin solution, castile soap, and benzalkonium chloride. All wounds received sharp debridement and irrigation with use of a pulsatile lavage device (19 psi). Images and photon counts were obtained prior to irrigation, after irrigation, and forty eight hours after injury and inoculation. For Study 2, we used the same animal model and compared bulb syringe and pulsatile lavage irrigation with saline solution. RESULTS: In Study 1, the irrigation treatment lowered the bacterial counts in all treatment groups. The greatest reduction was seen with castile soap, which lowered the photon count to 13% of the pretreatment level. This was followed by benzalkonium chloride, bacitracin, and saline solution at 18%, 22%, and 29%, respectively. At forty-eight hours, imaging showed a rebound in bacterial counts in every group. The highest rebound was measured in the castile soap group, which rebounded to 120% of the pretreatment level. The benzalkonium chloride group experienced a rebound to 94% of the pretreatment level. These were followed by bacitracin solution (89%) and normal saline solution (68%). In Study 2, both treatment methods were effective in removing 75% of the bacteria initially. At forty-eight hours, the bacterial levels in the pulsed lavage group rebounded to 94% of the original levels (compared with 48% in the bulb syringe group). The difference in the mean photon count ratios at forty-eight hours was significant (p = 0.048). CONCLUSIONS: Approaches used to remove bacteria from wounds, such as irrigants other than saline solution or high-pressure devices, may not have the best clinical outcome. PMID- 19122084 TI - Chondral injuries of the ankle with recurrent lateral instability: an arthroscopic study. AB - BACKGROUND: Prolonged lateral instability of the ankle after ligament injury has been believed to be a major cause of osteoarthritis of the ankle, yet the rate of development of osteoarthritis of the ankle is relatively low. Clarifying the relationship between patient factors and chondral damage of the ankle with prolonged instability is essential to identify the important risk factors underlying osteoarthritis of the ankle. METHODS: Arthroscopic examination was performed to assess the condition of the articular cartilage in a series of patients with prolonged lateral instability of the ankle. There were ninety-three patients with ninety-nine involved ankles. Their mean age was 28.7 years (range, fifteen to fifty-nine years). The relationships between the severity of the chondral damage and patient factors, the number and combination of torn ligaments, and mechanical instability and alignment of the ankle mortise were studied. RESULTS: Twenty-three ankles were classified as grade 0 (normal cartilage); thirty-five, as grade 1 (superficial softening, fibrillation, or fissuring of the cartilage); twenty-four, as grade 2 (a cartilage defect without exposure of the subchondral bone); and seventeen, as grade 3 (exposure of the subchondral bone). Patient age, the talar tilt angle, and varus inclination of the ankle plafond were significantly associated with more severe chondral changes. CONCLUSIONS: Patient age, the talar tilt angle, and varus inclination of the ankle are risk factors for severe chondral damage of the ankle in patients with a prolonged history of lateral ankle instability. PMID- 19122085 TI - Anterior-posterior and rotatory stability of single and double-bundle anterior cruciate ligament reconstructions. AB - BACKGROUND: Some surgeons presently reconstruct both the anteromedial and posterolateral bundles of the anterior cruciate ligament. The purposes of this study were to measure the abilities of single-bundle and anatomic double-bundle reconstructions to restore anteroposterior laxities and rotational kinematics to intact knee levels and to compare graft forces in reconstructed knees with forces in the native anterior cruciate ligament for the same loading conditions. METHODS: Native anterior cruciate ligament force and tibial rotations were recorded during passive knee extension tests with and without applied tibial loads. The anteromedial and posterolateral bundles were reconstructed with patellar tendon tissue sized to fit tightly within 7-mm femoral tunnels. Testing was repeated with the anteromedial graft alone (single bundle), tensioned to restore anteroposterior laxity at 30 degrees of flexion, and with double-bundle grafts. For double-bundle reconstructions, the anteromedial graft was first tensioned as above and then the posterolateral graft was tensioned with use of one of four protocols: posterolateral tension = anteromedial tension at 10 degrees (DB1), posterolateral tension = anteromedial tension at 30 degrees (DB2), posterolateral tension = (anteromedial tension + 30 N) at 10 degrees (DB3), and posterolateral tension = (anteromedial tension + 30 N) at 30 degrees (DB4). RESULTS: The posterolateral graft underwent a greater length change than the anteromedial graft between 0 degrees and 90 degrees . This difference in elongation patterns produced high forces in the posterolateral graft at 0 degrees when both grafts were tensioned and fixed at 30 degrees . The mean laxities for single-bundle reconstructions were within 1.1 mm of those of the intact knee between 0 degrees and 90 degrees ; the mean graft force at 0 degrees was 76 N. The mean laxities for DB4 reconstructions were from 0.9 to 2.8 mm less than those of the intact knee, and the mean graft force at 0 degrees was 264 N. Coupled internal tibial rotations from valgus moment were normal with the single-bundle graft. Internal rotations from tibial torque were approximately 2 degrees to 4 degrees greater than normal with a single-bundle graft. DB3 and DB4 reconstructions overcorrected the coupled tibial rotations from valgus moment and restored tibial rotations from internal torque to normal from 0 degrees to 45 degrees . The graft forces from tibial torque and valgus moment were normal with the single-bundle graft. The mean double-bundle graft forces at 0 degrees were 57 N to 143 N and 34 N to 171 N greater than normal for internal torque and valgus moment, respectively. CONCLUSIONS: The single-bundle reconstruction produced graft forces, knee laxities, and coupled tibial rotations that were closest to normal. Adding a posterolateral graft to an anteromedial graft tended to reduce laxities and tibial rotations, but the reductions were accompanied by markedly higher forces in the posterolateral graft near 0 degrees that occasionally caused it to fail during tests with internal torque or anterior tibial force. PMID- 19122086 TI - Revision arthroplasty with use of a reverse shoulder prosthesis-allograft composite. AB - BACKGROUND: Patients with disabling pain and loss of shoulder function with associated proximal humeral bone loss following shoulder arthroplasty have limited reliable treatment options. Our objective was to report the results, obtained as part of a prospective outcomes study, of the use of a reverse shoulder prosthesis-allograft composite in these patients. METHODS: Between 2002 and 2005, 353 patients treated with a reverse shoulder prosthesis were enrolled in a prospective cohort study. Twenty-five patients received, in addition, a proximal humeral allograft for the management of severe proximal humeral bone loss, and they comprise the study group. The average bone loss measured 53.6 mm (range, 34.5 to 150.3 mm). Patients were followed clinically with use of the American Shoulder and Elbow Surgeons (ASES) score, the Simple Shoulder Test (SST), and a scale with which the patients rated their satisfaction, and they were followed radiographically to detect mechanical failure, loosening, notching, and graft healing. All patients were followed for a minimum of two years (average, 30.2 months). RESULTS: The total average ASES score improved from 31.7 points preoperatively to 69.4 points at the time of follow-up (p < 0.0001), and the average SST score improved from 1.4 to 4.5 points (p < 0.0001). Nineteen patients (76%) reported a subjective good or excellent result, five reported a satisfactory result, and one reported that the result was unsatisfactory. The range of motion improved in forward flexion (from 32.7 degrees to 82.4 degrees ; p < 0.0001), abduction (from 40.4 degrees to 81.4 degrees ; p < 0.0001), and internal rotation. Radiographic evaluation at the time of final follow-up showed incorporation of the allograft in the metaphyseal region in 84% (twenty-one) of the twenty-five patients and incorporation of the allograft in the diaphyseal region in 76% (nineteen) of the patients. Four patients had complications. CONCLUSIONS: Use of a reverse shoulder prosthesis-proximal humeral allograft composite for the treatment of shoulder dysfunction following arthroplasty associated with substantial proximal humeral bone loss has shown promising early results. The allograft may restore proximal humeral bone stock, thereby helping to maintain the height of the prosthesis bone construct and thus deltoid tension. Additional, long-term studies are needed to evaluate the longevity of this construct. PMID- 19122087 TI - The epidemiology of revision total hip arthroplasty in the United States. AB - BACKGROUND: Understanding the causes of failure and the types of revision total hip arthroplasty performed is essential for guiding research, implant design, clinical decision-making, and health-care policy. The purpose of the present study was to evaluate the mechanisms of failure and the types of revision total hip arthroplasty procedures performed in the United States with use of newly implemented ICD-9-CM (International Classification of Diseases, Ninth Revision, Clinical Modification) diagnosis and procedure codes related specifically to revision total hip arthroplasty in a large, nationally representative population. METHODS: The Healthcare Cost and Utilization Project Nationwide Inpatient Sample database was used to analyze clinical, demographic, and economic data from 51,345 revision total hip arthroplasty procedures performed between October 1, 2005, and December 31, 2006. The prevalence of revision procedures was calculated for population subgroups in the United States that were stratified according to age, sex, diagnosis, census region, primary payer class, and type of hospital. The cause of failure, the average length of stay, and total charges were also determined for each type of revision arthroplasty procedure. RESULTS: The most common type of revision total hip arthroplasty procedure performed was all component revision (41.1%), and the most common causes of revision were instability/dislocation (22.5%), mechanical loosening (19.7%), and infection (14.8%). Revision total hip arthroplasty procedures were most commonly performed in large, urban, nonteaching hospitals for Medicare patients seventy-five to eighty-four years of age. The average length of hospital stay for all types of revision arthroplasties was 6.2 days, and the average total charges were $54,553. However, the average length of stay, average charges, and procedure frequencies varied considerably according to census region, hospital type, and type of revision total hip arthroplasty procedure performed. CONCLUSIONS: Hip instability and mechanical loosening are the most common indications for revision total hip arthroplasty in the United States. As further experience is gained with the new diagnosis and procedure codes specifically related to revision total hip arthroplasty, this information will be valuable in directing future research, implant design, and clinical decision-making. PMID- 19122088 TI - The Knee Arthroplasty Trial (KAT) design features, baseline characteristics, and two-year functional outcomes after alternative approaches to knee replacement. AB - BACKGROUND: The aim of continued development of total knee replacement systems has been the further improvement of the quality of life and increasing the duration of prosthetic survival. Our goal was to evaluate the effects of several design features, including metal backing of the tibial component, patellar resurfacing, and a mobile bearing between the tibial and femoral components, on the function and survival of the implant. METHODS: A pragmatic, multicenter, randomized, controlled trial involving 116 surgeons in thirty-four centers in the United Kingdom was performed; 2352 participants were randomly allocated to be treated with or without a metal backing of the tibial component (409), with or without patellar resurfacing (1715), and/or with or without a mobile bearing (539). Randomization to more than one comparison was allowed. The primary outcome measures were the Oxford Knee Score (OKS), Short Form-12, EuroQol-5D, and the need for additional surgery. The results up to two years postoperatively are reported. RESULTS: Functional status and quality-of-life scores were low at baseline but improved markedly across all trial groups following knee replacement (mean overall OKS, 17.98 points at baseline and 34.82 points at two years). Most of the change was observed at three months after the surgery. Six percent of the patients had additional knee surgery within two years. There was no evidence of differences in clinical, functional, or quality-of-life measures between the randomized groups at two years. CONCLUSIONS: Patients have substantial improvement following total knee replacement. This is the first adequately powered randomized controlled trial, of which we are aware, in which the effects of metal backing, patellar resurfacing, and a mobile bearing were investigated. We found no evidence of an effect of these variants on the rate of early complications or on functional recovery up to two years after total knee replacement. PMID- 19122089 TI - Ipsilateral femoral autograft reconstruction after resection of a pelvic tumor. AB - BACKGROUND: Reconstruction of bone after the resection of a pelvic tumor is challenging. The purpose of the present study was to evaluate the use of the ipsilateral femur as the graft material for reconstruction. METHODS: We performed a retrospective review of thirteen patients with a malignant pelvic lesion who underwent resection followed by reconstruction with an ipsilateral femoral autograft and insertion of a total hip replacement. The study group included nine men and four women with a median age of fifty-one years at the time of the reconstruction. The diagnosis was chondrosarcoma in eight patients, metastasis in three, and myeloma and radiation-induced malignant disease in one each. The surviving patients were assessed functionally and radiographically; the cumulative probability of revision was estimated while taking into account competing risks. RESULTS: The median duration of follow-up was forty-nine months. At the time of the latest follow-up, seven patients were alive and disease-free and six had died from metastatic disease. Four patients had had revision of the reconstruction, two for the treatment of mechanical complications and two for the treatment of infection. Three other patients had mechanical complications but had not had a revision. The cumulative probability of revision of the reconstruction for mechanical failure was 8% (95% confidence interval, 0% to 23%), 8% (95% confidence interval, 0% to 23%), and 16% (95% confidence interval, 0% to 39%) at one, two, and four years, respectively. CONCLUSIONS: Although it has attendant complications consistent with pelvic tumor surgery, an ipsilateral femoral autograft reconstruction may be an option for reconstruction of pelvic discontinuity in a subgroup of patients following tumor resection. This innovative procedure requires longer-term follow-up studies. PMID- 19122090 TI - Fixator-assisted acute femoral deformity correction and consecutive lengthening over an intramedullary nail. AB - BACKGROUND: External fixators are being used frequently in standard limb lengthening and deformity-correction procedures. Lengthening over an intramedullary nail has been a successful technique, and fixator-assisted intramedullary nailing has provided satisfactory results for the correction of selected deformities. We report a combined technique for the treatment of femoral deformities associated with shortening. METHODS: Between 1997 and 2005, twenty eight femora in twenty-five patients with a median age of twenty-seven years underwent reconstruction with an intramedullary nail and a unilateral fixator. The mean amount of shortening was 6.33 cm, and the mean preoperative mechanical axis deviation was 33.86 mm. Deformity correction was performed acutely and secured by the intramedullary nail, which was locked distally, and the same external fixator that was used for the deformity correction was utilized for lengthening. At the end of the distraction period, proximal locking screws were placed in the intramedullary nail and the external fixator was removed. At the time of follow-up, deformity correction and bone healing were assessed clinically and radiographically, complications were noted, and the functional results were assessed. RESULTS: The mean duration of follow-up was forty months. The mean duration of the external fixation was 83.29 days, and the mean external fixation index was 14.98 days/cm. The mean amount of lengthening was 6.02 cm. The mean amount of mechanical axis deviation at the end of the treatment was 11.29 mm. The mean bone healing index was 36.66 days/cm. A knee flexion contracture developed in one patient and resolved after intensive rehabilitation. One patient underwent two revisions because of Schanz screw displacement secondary to cortical fracture, and four patients with minor pin-track infections were treated successfully with local wound care and oral antibiotics. CONCLUSIONS: While femoral lengthening and deformity correction can be obtained with classic methods for application of an external fixator, the long period of external fixation, patient discomfort, and plastic deformation of the regenerated bone after removal of the fixator are major disadvantages. Two techniques, fixator-assisted nailing and lengthening over an intramedullary nail, were combined in this series. The duration of the external fixation was reduced compared with that required for classic treatment with an external fixator and patient comfort was increased. In addition, the intramedullary nail prevented fracture and deformation of the regenerated bone. PMID- 19122091 TI - Long-term results of cemented metal-backed glenoid components for osteoarthritis of the shoulder. AB - BACKGROUND: There has been recent renewed interest in the use of metal-backed glenoid components in total shoulder arthroplasty. However, little information is available on the long-term results. The purpose of this study was to determine the results, the risk factors for an unsatisfactory outcome, and the failure rates of total shoulder arthroplasty with a cemented metal-backed glenoid component. METHODS: Between 1985 and 1991, 100 total shoulder arthroplasties with Neer cemented metal-backed glenoid components were performed at our institution to treat osteoarthritis. Ninety-five shoulders were followed for a minimum of two years (mean, 10.8 years) or until the time of revision surgery. Patients were assessed with use of a modified Neer rating system at the time of the latest follow-up. Radiographs of eighty-three shoulders were assessed for the presence of glenoid erosion, glenohumeral subluxation, periprosthetic lucency, and a shift in component position. RESULTS: Total shoulder arthroplasty with a cemented metal backed glenoid component was significantly associated with pain relief (p < 0.001) as well as with an improvement in abduction from a mean of 92 degrees to 146 degrees (p < 0.001) and external rotation from a mean of 26 degrees to 60 degrees (p < 0.001). According to the modified Neer result-rating system, the result was excellent in forty-seven shoulders, satisfactory in twenty-seven shoulders, and unsatisfactory in twenty-one shoulders. Five patients underwent revision surgery because of component loosening (two patients), component subluxation (one patient), a fracture distal to the stem (one patient), and polyethylene wear (one patient). Glenoid periprosthetic lucency was present in sixty-nine (83%) of eighty-three shoulders at a minimum radiographic follow-up of two years. CONCLUSIONS: The data from this study suggest that total shoulder arthroplasty with a cemented metal-backed glenoid component for the treatment of osteoarthritis is associated with pain relief and improvement in motion. However, the high rate of glenoid periprosthetic lucency is concerning and requires additional follow-up and investigation. These results are not better than those that have been reported for all-polyethylene cemented glenoid components. PMID- 19122092 TI - Analysis of tibiofemoral cartilage deformation in the posterior cruciate ligament deficient knee. AB - BACKGROUND: Degeneration of the tibiofemoral articular cartilage often develops in patients with posterior cruciate ligament deficiency, yet little research has focused on the etiology of this specific type of cartilage degeneration. In this study, we hypothesized that posterior cruciate ligament deficiency changes the location and magnitude of cartilage deformation in the tibiofemoral joint. METHODS: Fourteen patients with a posterior cruciate ligament injury in one knee and the contralateral side intact participated in the study. First, both knees were imaged with use of a specific magnetic resonance imaging sequence to create three-dimensional knee models of the surfaces of the bone and cartilage. Next, each patient performed a single leg lunge as images were recorded with a dual fluoroscopic system at 0 degrees, 30 degrees, 60 degrees, 75 degrees, 90 degrees, 105 degrees, and 120 degrees of knee flexion. Finally, the three-dimensional knee models and fluoroscopic images were used to reproduce the in vivo knee position at each flexion angle with use of a previously described image-matching method. With use of these series of knee models, the location and magnitude of peak tibiofemoral cartilage deformation at each flexion angle were compared between the intact contralateral and posterior cruciate ligament-deficient knees. RESULTS: In the medial compartment of the posterior cruciate ligament-deficient knees, the location and magnitude of peak cartilage deformation were significantly changed, compared with those in the intact contralateral knees, between 75 degrees and 120 degrees of flexion, with a more anterior and medial location of peak cartilage deformation on the tibial plateau as well as increased deformation of the cartilage. In the lateral compartment, no significant differences in the location or magnitude of peak cartilage deformation were found between the intact and posterior cruciate ligament-deficient knees. CONCLUSIONS: The altered kinematics associated with posterior cruciate ligament deficiency resulted in a shift of the tibiofemoral contact location and an increase in cartilage deformation in the medial compartment beyond 75 degrees of knee flexion. The magnitude of the medial contact shift in the posterior cruciate ligament-deficient knee was on the same order as that of the anterior contact shift. PMID- 19122093 TI - Architectural analysis and intraoperative measurements demonstrate the unique design of the multifidus muscle for lumbar spine stability. AB - BACKGROUND: Muscular instability is an important risk factor for lumbar spine injury and chronic low-back pain. Although the lumbar multifidus muscle is considered an important paraspinal muscle, its design features are not completely understood. The purpose of the present study was to determine the architectural properties, in vivo sarcomere length operating range, and passive mechanical properties of the human multifidus muscle. We hypothesized that its architecture would be characterized by short fibers and a large physiological cross-sectional area and that it would operate over a relatively wide range of sarcomere lengths but would have very stiff passive material properties. METHODS: The lumbar spines of eight cadaver specimens were excised en bloc from T12 to the sacrum. Multifidus muscles were isolated from each vertebral level, permitting the architectural measurements of mass, sarcomere length, normalized fiber length, physiological cross-sectional area, and fiber length-to-muscle length ratio. To determine the sarcomere length operating range of the muscle, sarcomere lengths were measured from intraoperative biopsy specimens that were obtained with the spine in the flexed and extended positions. The material properties of single muscle fibers were obtained from passive stress-strain tests of excised biopsy specimens. RESULTS: The average muscle mass (and standard error) was 146 +/- 8.7 g, and the average sarcomere length was 2.27 +/- 0.06 microm, yielding an average normalized fiber length of 5.66 +/- 0.65 cm, an average physiological cross sectional area of 23.9 +/- 3.0 cm(2), and an average fiber length-to-muscle length ratio of 0.21 +/- 0.03. Intraoperative sarcomere length measurements revealed that the muscle operates from 1.98 +/- 0.15 microm in extension to 2.70 +/- 0.11 microm in flexion. Passive mechanical data suggested that the material properties of the muscle are comparable with those of muscles of the arm or leg. CONCLUSIONS: The architectural design (a high cross-sectional area and a low fiber length-to-muscle length ratio) demonstrates that the multifidus muscle is uniquely designed as a stabilizer to produce large forces. Furthermore, multifidus sarcomeres are positioned on the ascending portion of the length tension curve, allowing the muscle to become stronger as the spine assumes a forward-leaning posture. PMID- 19122094 TI - Tuberculous arthritis of the knee treated with two-stage total knee arthroplasty. A case report. PMID- 19122095 TI - Anterior dislocation of the hip after arthroscopy in a patient with capsular laxity of the hip. A case report. PMID- 19122096 TI - A comprehensive review of partial anterior cruciate ligament tears. AB - A partial tear of the anterior cruciate ligament is characterized by an asymmetric Lachman-test result, a negative pivot-shift test, a low-grade KT-1000 arthrometer measurement (< or =3 mm), and arthroscopic evidence of anterior cruciate ligament injury. The pivot shift test is the most important test in determining anterior cruciate ligament insufficiency. A positive test, independent of the grade, is indicative of a functionally deficient anterior cruciate ligament. Nonoperative management results in an acceptable clinical outcome in the majority of cases. Progression to knee instability (anterior translation and rotation) depends on the extent of the anterior cruciate ligament injury and the activity level of the patient. Thermal treatment is not recommended for partial tears of the anterior cruciate ligament. Symptomatic instability is treated reliably with anterior cruciate ligament reconstruction. PMID- 19122098 TI - Maintenance of certification for orthopaedic surgeons. PMID- 19122097 TI - The young adult with hip impingement: deciding on the optimal intervention. PMID- 19122099 TI - Economic viability of a community-based level-II orthopaedic trauma system. PMID- 19122100 TI - Current hand surgery literature as an educational tool for the orthopaedic in training examination. PMID- 19122101 TI - What's new in sports medicine. PMID- 19122102 TI - MYB58 and MYB63 are transcriptional activators of the lignin biosynthetic pathway during secondary cell wall formation in Arabidopsis. AB - It has previously been shown that SECONDARY WALL-ASSOCIATED NAC DOMAIN PROTEIN1 (SND1) is a key transcription factor regulating secondary cell wall formation, including the biosynthesis of cellulose, xylan, and lignin. In this study, we show that two closely related SND1-regulated MYB transcription factors, MYB58 and MYB63, are transcriptional regulators specifically activating lignin biosynthetic genes during secondary wall formation in Arabidopsis thaliana. MYB58 and MYB63 are phylogenetically distinct from previously characterized MYBs shown to be associated with secondary wall formation or phenylpropanoid metabolism. Expression studies showed that MYB58 and MYB63 are specifically expressed in fibers and vessels undergoing secondary wall thickening. Dominant repression of their functions led to a reduction in secondary wall thickening and lignin content. Overexpression of MYB58 and MYB63 resulted in specific activation of lignin biosynthetic genes and concomitant ectopic deposition of lignin in cells that are normally unlignified. MYB58 was able to activate directly the expression of lignin biosynthetic genes and a secondary wall-associated laccase (LAC4) gene. Furthermore, the expression of MYB58 and MYB63 was shown to be regulated by the SND1 close homologs NST1, NST2, VND6, and VND7 and their downstream target MYB46. Together, our results indicate that MYB58 and MYB63 are specific transcriptional activators of lignin biosynthesis in the SND1-mediated transcriptional network regulating secondary wall formation. PMID- 19122103 TI - A polypyrimidine tract binding protein, pumpkin RBP50, forms the basis of a phloem-mobile ribonucleoprotein complex. AB - RNA binding proteins (RBPs) are integral components of ribonucleoprotein (RNP) complexes and play a central role in RNA processing. In plants, some RBPs function in a non-cell-autonomous manner. The angiosperm phloem translocation stream contains a unique population of RBPs, but little is known regarding the nature of the proteins and mRNA species that constitute phloem-mobile RNP complexes. Here, we identified and characterized a 50-kD pumpkin (Cucurbita maxima cv Big Max) phloem RNA binding protein (RBP50) that is evolutionarily related to animal polypyrimidine tract binding proteins. In situ hybridization studies indicated a high level of RBP50 transcripts in companion cells, while immunolocalization experiments detected RBP50 in both companion cells and sieve elements. A comparison of the levels of RBP50 present in vascular bundles and phloem sap indicated that this protein is highly enriched in the phloem sap. Heterografting experiments confirmed that RBP50 is translocated from source to sink tissues. Collectively, these findings established that RBP50 functions as a non-cell-autonomous RBP. Protein overlay, coimmunoprecipitation, and cross linking experiments identified the phloem proteins and mRNA species that constitute RBP50-based RNP complexes. Gel mobility-shift assays demonstrated that specificity, with respect to the bound mRNA, is established by the polypyrimidine tract binding motifs within such transcripts. We present a model for RBP50-based RNP complexes within the pumpkin phloem translocation stream. PMID- 19122105 TI - Ureaplasma parvum or Mycoplasma hominis as sole pathogens cause chorioamnionitis, preterm delivery, and fetal pneumonia in rhesus macaques. AB - The authors assess causal, cellular and inflammatory links between intraamniotic infection with Ureaplasma parvum or Mycoplasma hominis and preterm labor in a nonhuman primate model. Long-term catheterized rhesus monkeys received intraamniotic inoculations of clinical isolates of Ureaplasma parvum serovar 1, M hominis, media control or physiological saline. Genital mycoplasmas were quantified in amniotic fluid (AF) and documented in fetal tissues by culture and PCR. In association with elevated AF colony counts for U parvum or M hominis, there was a sequential upregulation of AF leukocytes, proinflammatory cytokines, prostaglandin E2 and F2a, metalloproteinase-9 and uterine activity ( P< .05). Fetal membranes and lung were uniformly positive for both microorganisms; fetal blood and cerebrospinal fluid cultures and PCR were more often positive for M hominis than U parvum. Histopathologic findings of chorioamnionitis, a systemic fetal inflammatory response and pneumonitis worsen with duration of in utero infection. U parvum or M hominis, as sole pathogens, elicit a robust proinflammatory response which contributes to preterm labor and fetal lung injury. PMID- 19122104 TI - SQUAMOSA Promoter Binding Protein-Like7 Is a Central Regulator for Copper Homeostasis in Arabidopsis. AB - Expression of miR398 is induced in response to copper deficiency and is involved in the degradation of mRNAs encoding copper/zinc superoxide dismutase in Arabidopsis thaliana. We found that SPL7 (for SQUAMOSA promoter binding protein like7) is essential for this response of miR398. SPL7 is homologous to Copper response regulator1, the transcription factor that is required for switching between plastocyanin and cytochrome c(6) in response to copper deficiency in Chlamydomonas reinhardtii. SPL7 bound directly to GTAC motifs in the miR398 promoter in vitro, and these motifs were essential and sufficient for the response to copper deficiency in vivo. SPL7 is also required for the expression of multiple microRNAs, miR397, miR408, and miR857, involved in copper homeostasis and of genes encoding several copper transporters and a copper chaperone, indicating its central role in response to copper deficiency. Consistent with this idea, the growth of spl7 plants was severely impaired under low-copper conditions. PMID- 19122106 TI - Surprises and reaffirmations in 2008 clinical trials. PMID- 19122107 TI - A persistently swollen lip. PMID- 19122108 TI - Hepatitis B: a strategy for evaluation and management. AB - In hepatitis B virus (HBV) infection, a single approach to treatment cannot be applied to all patients. Acute, adult-acquired HBV infection rarely requires treatment, whereas treatment for chronic infection should be based on the patient's clinical situation and test results. The ideal agent for treating hepatitis B does not exist, and trade-offs are the essence of agent selection. In last month's Journal (Cleve Clin J Med 2008; 75:881-889), we outlined the natural history and diagnosis of chronic HBV infection; in this article we outline its management. PMID- 19122110 TI - Q: Who should receive the shingles vaccine? PMID- 19122111 TI - Alzheimer disease: time to improve its diagnosis and treatment. AB - Basic research is bringing a much-needed infusion of optimism and urgency to the clinical diagnosis and treatment of Alzheimer disease. Some of its risk factors may be modifiable, and although current drugs offer only modest benefit, true disease-modifying drugs are on the horizon. This review is aimed at primary care physicians, who are the first clinicians to see patients with Alzheimer disease and are responsible for their ongoing care throughout the course of their dementia. PMID- 19122109 TI - Jupiter to earth: a statin helps people with normal LDL-C and high hs-CRP, but what does it mean? AB - The JUPITER trial (Justification for the Use of Statins in Prevention: an Intervention Trial Evaluating Rosuvastatin) (N Engl J Med 2008; 359:2195-2207) compared rosuvastatin (Crestor) 20 mg daily vs placebo in apparently healthy people who had levels of low-density lipoprotein cholesterol (LDL-C) lower than 130 mg/dL but elevated levels (>or= 2 mg/L) of high-sensitivity C-reactive protein (hs-CRP). Rosuvastatin treatment lowered LDL-C levels by 50% and hs-CRP levels by 37%, accompanied by a 44% relative risk reduction in the composite end point of unstable angina, revascularization, and confirmed death from cardiovascular causes. In absolute terms, 95 people had to be treated over 2 years to prevent one event. There was, however, a higher incidence of diabetes in the rosuvastatin group. PMID- 19122112 TI - Depression and heart disease: what do we know, and where are we headed? AB - Depression and heart disease have an intricate association and perhaps a causal relationship. We review the current status of depression and heart disease and provide an algorithm for diagnosing and treating depression in cardiac patients that internists and cardiologists can use in their daily patient encounters. PMID- 19122117 TI - More frequent Blood transfusions. PMID- 19122113 TI - Mice selectively bred for high- or low-alcohol-induced locomotion exhibit differences in dopamine neuron function. AB - Elevated sensitivity to the euphoric or stimulant effects of ethanol is associated with higher levels of alcohol use in some human populations. Midbrain dopamine neurons are thought to be important mediators of both ethanol reward and locomotor stimulation. Patch-clamp recordings were used to examine the electrical properties of dopamine neurons in a genetic model of heightened (FAST) and reduced (SLOW) sensitivity to the locomotor-activating effects of ethanol. Pacemaker firing of dopamine neurons was faster in FAST than SLOW mice, as was the current density through I(H) channels. Acute administration of ethanol accelerated the firing of dopamine neurons to a greater extent in recordings from FAST than SLOW mice. Dopamine neurons from FAST mice also exhibited reduced GABA(A) receptor-mediated synaptic input, compared with SLOW mice. The results suggest that dopamine neuron I(H) channels, firing rate, and GABAergic input may play a role in sensitivity to the locomotor activation observed at early time points after ethanol administration and could underlie differences in sensitivity to alcohol relevant to risk for alcohol abuse. PMID- 19122118 TI - Dendritic cells differentiate into osteoclasts in bone marrow microenvironment in vivo. PMID- 19122120 TI - Differential gene expression in central nervous system lymphoma. PMID- 19122122 TI - Aftermath of suicide in the hospital: institutional response. AB - BACKGROUND: A suicide can be a devastating event in the hospital, and few guidelines exist to aid an institution's response. METHOD: The authors describe a framework of immediate, short-term and long-term responses in the event of an in hospital suicide. CONCLUSION: Implications for administration, communication, assessment, physical environment, and standards of care throughout the hospital are discussed that are relevant to both general-medical and psychiatric settings. Suggestions for the successful management of the aftermath of a suicide, such as the formation of a multidisciplinary leadership team, are included. PMID- 19122123 TI - Customizing treatment of chronic fatigue syndrome and fibromyalgia: the role of perpetuating factors. AB - BACKGROUND: Syndromes characterized by chronic, medically unexplained fatigue, effort- and stress-intolerance, and widespread pain are highly prevalent in medicine. RESULTS: In chronic fatigue syndrome (CFS) and fibromyalgia (FM), various perpetuating factors may impair patients' quality of life and functioning and impede recovery. Although cognitive-behavioral and graded-exercise therapy are evidence-based treatments, the effectiveness and acceptability of therapeutic interventions in CFS/FM may largely depend on a customized approach taking the heterogeneity of perpetuating factors into account. CONCLUSION: Further research should clarify the aim and outcome of different treatment strategies in CFS/FM, as well as the underlying mechanisms of change, including those facilitating neurobiological recovery. PMID- 19122124 TI - Impact of antidepressant use on gastric bypass surgery patients' weight loss and health-related quality-of-life outcomes. AB - BACKGROUND: Psychological factors may be predictive of post-surgical adjustment and successful outcomes in obesity-surgery populations. OBJECTIVE: The authors compared post-gastric bypass surgery (GBS) outcomes of patients being given active psychotropic treatment for depression, with those of patients without treatment or identifiable need for treatment at enrollment. METHOD: Outcome measures included weight and repeated administration of the Medical Outcome Survey Short Form-36 (SF-36). RESULTS: There was no impact of antidepressant treatment on GBS patients' weight loss or SF-36 outcomes. CONCLUSION: Therefore, patients with actively treated depression should be expected to have GBS outcomes equivalent to those of patients without identifiable psychiatric illness or treatment. PMID- 19122125 TI - Toward defining the scope of psychosomatic medicine practice: psychosomatic medicine in an outpatient, tertiary-care practice setting. AB - BACKGROUND: Because psychosomatic medicine (PM) is increasingly practiced in outpatient settings, the scope of practice needs to be delineated from community psychiatry and inpatient psychiatry work. OBJECTIVE: The authors sought to address the question of whether outpatient activities are a definably part the scope of practice of PM. METHOD: Three clinical groups were compared: 200 PM outpatients, 200 consultation-liaison (CL) inpatients, and 200 community psychiatry (CP) outpatients. RESULTS: The groups differed significantly in 49 of 112 demographic and clinical comparisons (43.8%). Analysis of individual measures validated the concept that PM outpatient practice requires traditional PM/CL expertise with medical-psychiatric differential diagnosis, unexplained physical symptoms, pain, and psychopharmacological management in medically ill and geriatric patients. CONCLUSION: Outpatient PM experiences may also enhance training opportunities, given an expanded case-mix. PMID- 19122126 TI - Psychosocial issues in hepatitis C: a qualitative analysis. AB - BACKGROUND: Despite demonstrated prevalence of psychosocial problems among hepatitis C (HCV) patients, little is known about how these problems develop, why they are sustained, and how clinical providers can effectively intervene. OBJECTIVE: The authors used a qualitative approach to investigate the nature of psychosocial issues in HCV. METHOD: Focus groups were conducted with HCV patients. RESULTS: Participants discussed significant feelings of anger, depression, and stigma associated with the diagnosis and noted that these feelings/experiences isolated them from potential sources of social support. CONCLUSION: Results have important implications for clinical providers and suggest pathways by which HCV-related psychosocial factors may interact with and affect quality of life. PMID- 19122127 TI - Migraine predicts self-reported muscle tension in patients with major depressive disorder. AB - BACKGROUND/OBJECTIVE: The aim of this study was to identify factors related to muscle tension in patients with major depressive disorder (MDD) with comorbid anxiety and migraine. METHOD: Consecutive psychiatric outpatients with MDD were enrolled. Self-reported muscle tension (SMT) during the previous week was evaluated with a 0-10 scale. RESULTS: Of 135 participants with MDD, 63 (46.7%) had migraine. Multiple-regression analyses showed that migraine and headache intensity were two major independent factors related to SMT. CONCLUSION: Further studies on musculoskeletal symptoms in MDD should not neglect the impact of migraine. PMID- 19122128 TI - Family caregivers and guilt in the context of cancer care. AB - BACKGROUND: Guilt as a key emotional phenomenon in the cancer-caregiving experience is an understudied issue. OBJECTIVE: The purpose of this study is to identify demographic characteristics of cancer caregivers and care-related stress factors that are associated with their feelings of caregiver guilt, as well as to explore the effect of caregiver guilt on their adjustment outcomes. METHOD: A total of 739 caregivers of cancer survivors completed a survey (66.7% response rate), of which 635 provided complete data for the measures in this study. RESULTS: Hierarchical regression analyses revealed that certain caregiver demographics (i.e., younger age, adult offspring, employed) and care-related stress factors (i.e., greater impact on schedule, less perceived caregiving competence, poorer overall health of the care-recipient) were significantly related to caregiver guilt. Higher levels of psychological distress and poorer mental, social, and physical functioning were significantly associated with caregiver guilt, above and beyond the variance accounted for by the covariates. CONCLUSION: Results suggest that caregiver guilt compromises the psychosocial and somatic adjustment of cancer caregivers. Guilt may be a cardinal feature of the caregiving experience, and to fully understand the implications of this complex phenomenon, more research is needed. PMID- 19122129 TI - Prevalence, characteristics, and attribution of somatic symptoms in Spanish patients with major depressive disorder seeking primary health care. AB - BACKGROUND: Somatic symptoms (SS) tend to dominate clinical symptomatology in patients with depression in primary care. OBJECTIVE: The authors performed a cross-sectional nationwide epidemiological study on 1,150 primary-care patients with major depression and evaluated the prevalence of SS and physicians' attribution of their origin. METHOD: Patients were administered the Structured Polyvalent Psychiatric Interview. RESULTS: Ninety-three percent of patients had at least one SS fully or partially attributed to depression, and 45% of patients had four to nine. Painful symptoms, despite being the most frequent, were the least often attributed to depression (fewer than 25% of patients with pain) and significantly more often attributed to a combined origin. CONCLUSION: Results suggest that primary-care physicians tend to associate pain with depression to a significantly lesser extent than any other somatic symptom (e.g., cardiopulmonary or gastrointestinal). Therefore, special attention should be given to painful symptoms in order to ensure efficient management of depression in primary care. PMID- 19122130 TI - Early-onset versus late-onset HIV-related secondary mania in Uganda. AB - BACKGROUND: First-episode secondary mania in human immunodeficiency virus (HIV) infection has been described among samples of predominantly Caucasian, HIV positive male patients in developed countries. OBJECTIVE: The goal of this study was to compare the demographic and clinical characteristics of HIV-positive patients with early-onset and late-onset first-episode secondary mania in HIV infection. There were previous findings of an association between late-onset mania and severe cognitive impairment. METHOD: Subjects were HIV inpatients with clinically-confirmed mania, who received standard demographic, psychiatric, physical, and laboratory assessments. Early-onset patients had CD4 cell counts > 200 mm(3); late-onset patients had CD4 cell counts < or = 200 mm(3). RESULTS: There were no demographic or cognitive differences between early-onset and late onset mania patients, and high rates of psychotic symptoms in both groups. However, late-onset patients had more manic symptoms. CONCLUSION: Late-onset HIV mania patients had more severe psychopathology and, thus, demonstrated a greater need for highly active retroviral therapy. PMID- 19122131 TI - Analysis of transfers from a medical-psychiatry inpatient unit to a medical surgical unit within 48 hours of admission. AB - BACKGROUND: Clinical screening of patients being presented for admission to a medical-psychiatric unit (MPU) is important to ensure safe, timely, and effective treatment. OBJECTIVE: Authors determined demographic and clinical characteristics of patients accepted for admission to a private MPU and who required transfer to medical-surgical units within 48 hours of admission. METHOD: Medical records of 1,583 consecutive admissions to the MPU were reviewed. The charts of patients transferred to a higher level of medical or surgical care within 48 hours were evaluated to categorize the reasons for transfers, and these were assessed for preventability. RESULTS: Thirty patients (1.9%) required transfer to a medical surgical unit. Reasons for transfer included pulmonary, cardiovascular, endocrine, and neurological etiologies. Most transfers (66.6%) were not foreseeable because symptom-onset was after admission. CONCLUSION: Inappropriate MPU admissions may have been avoided by more vigilant screening for pulmonary, cardiovascular, electrolyte, and infectious disorders. PMID- 19122132 TI - Panic as a harbinger of pancreatic cancer. AB - BACKGROUND: Previous literature has discussed the diagnosis of cancer with psychiatric symptoms. There are noted associations made between pancreatic cancer and depression and anxiety symptoms both in literature and textbooks. METHOD: The authors discuss the case of a patient who presented with anxiety and panic attacks before his diagnosis of pancreatic cancer. CONCLUSION: There has been a previous association noted between depression and anxiety and pancreatic cancer; however, this work and another report have noted panic episodes preceding diagnosis, as well. These symptoms could prove to be an important marker for early detection. PMID- 19122133 TI - Obsessive-compulsive disorder with mitochondrial disease. AB - BACKGROUND: Mitochondrial diseases are among the most common genetic disorders, and they have been associated with several psychiatric syndromes. METHOD: The authors present two cases of obsessive-compulsive disorder (OCD) occurring in patients with MELAS (the A3243G mutation). RESULTS: Their clinical course and response to standard OCD treatment strategies was poor. DISCUSSION: Possible mechanisms for OCD symptoms are suggested by animal models and neuropathological findings. It remains unclear whether different types of mitochondrial disorders are associated with particular neuropsychiatric symptoms. Psychiatric symptoms may predate the diagnosis of mitochondrial disorder; thus, psychiatrists should consider mitochondrial disorders in the presence of common physical signs that are typically associated with these disorders. PMID- 19122134 TI - Non-fearful panic disorder in gastroenterology. AB - BACKGROUND: Nonfearful panic disorder (NFPD) is a panic condition masked under the appearance of somatic symptoms only, without the component of fear, and it represents a challenging diagnostic task. METHOD: This is the first case report of NFPD in a male patient with acute gastric pain and gastrointestinal disease (atrophic gastritis and H. pylori infection). RESULTS: The patient showed atypical panic symptoms and demoralization on the Diagnostic Criteria for Psychosomatic Research screening. He was successfully treated with anti-panic medication and cognitive-behavioral therapy. DISCUSSION: The case report shows that accurate psychosomatic assessment may help clinicians avoid diagnostic delay, prevent the administration of unnecessary medications, and give patients more appropriate treatment. PMID- 19122135 TI - Successful use of clozapine with immunosuppressive therapy in a renal-transplant patient. PMID- 19122136 TI - Chronic obstructive pulmonary disease and panic disorder: their interrelationships and a unique utilization of beta-receptor agonists. PMID- 19122138 TI - Directed self-organization of single DNA molecules in a nanoslit via embedded nanopit arrays. AB - We show that arrays of nanopit structures etched in a nanoslit can control the positioning and conformation of single DNA molecules in nanofluidic devices. By adjusting the spacing, organization and placement of the nanopits it is possible to immobilize DNA at predetermined regions of a device without additional chemical modification and achieve a high degree of control over local DNA conformation. DNA can be extended between two nanopits and in closely spaced arrays will self-assemble into "connect-the-dots" conformations consisting of locally pinned segments joined by fluctuating linkers. These results have broad implications for nanotechnology fields that require methods for the nanoscale positioning and manipulation of DNA. PMID- 19122139 TI - Ebf1-mediated down-regulation of Id2 and Id3 is essential for specification of the B cell lineage. AB - Gene knockout experiments in mice have suggested a hierarchical model of early B cell commitment wherein E2A proteins (E47 and E12) activate early B cell factor (Ebf1), which in turn activates expression of the B cell commitment factor, Pax5. In IL-7 receptor alpha (IL-7Ralpha) knockout mice, B cell development is blocked before B-lineage commitment at the prepro-B cell stage in adult animals. In IL 7Ralpha(-/-) prepro-B cells, E47 is expressed and yet is insufficient to transcriptionally activate the putative downstream target gene, Ebf1. In this study, we show that further increases of E47 expression in IL-7Ralpha(-/-) prepro B cells fails to activate Ebf1, but rather leads to a dramatic induction of the E2A inhibitory factors, Id2 and Id3. In contrast, enforced expression of Ebf1 in IL-7Ralpha(-/-) bone marrow potently down-regulates Id2 and Id3 mRNA expression and restores B cell differentiation in vivo. Down-regulation of both Id2 and Id3 during B cell specification is essential in that overexpression of either Id2 or Id3 in wild-type bone marrow blocks B cell specification at the prepro-B cell stage. Collectively, these studies suggest a model where Ebf1 induction specifies the B cell fate by dramatically increasing activity of E47 at the posttranslational level. PMID- 19122140 TI - Low-barrier hydrogen bond in photoactive yellow protein. AB - Low-barrier hydrogen bonds (LBHBs) have been proposed to play roles in protein functions, including enzymatic catalysis and proton transfer. Transient formation of LBHBs is expected to stabilize specific reaction intermediates. However, based on experimental results and theoretical considerations, arguments against the importance of LBHB in proteins have been raised. The discrepancy is caused by the absence of direct identification of the hydrogen atom position. Here, we show by high-resolution neutron crystallography of photoactive yellow protein (PYP) that a LBHB exists in a protein, even in the ground state. We identified approximately 87% (819/942) of the hydrogen positions in PYP and demonstrated that the hydrogen bond between the chromophore and E46 is a LBHB. This LBHB stabilizes an isolated electric charge buried in the hydrophobic environment of the protein interior. We propose that in the excited state the fast relaxation of the LBHB into a normal hydrogen bond is the trigger for photo-signal propagation to the protein moiety. These results give insights into the novel roles of LBHBs and the mechanism of the formation of LBHBs. PMID- 19122141 TI - HIV-1 Tat RNA silencing suppressor activity is conserved across kingdoms and counteracts translational repression of HIV-1. AB - The RNA silencing pathway is an intracellular innate response to virus infections and retro-transposons. Many plant viruses counter this host restriction by RNA silencing suppressor (RSS) activity of a double-stranded RNA-binding protein, e.g., tomato bushy stunt virus P19. Here, we demonstrate P19 and HIV-1 Tat function across the plant and animal kingdoms and suppress a common step in RNA silencing that is downstream of small RNA maturation. Our experiments reveal that RNA silencing in HIV-1 infected human cells severely attenuates the translational output of the unspliced HIV-1 gag mRNA, and possibly all HIV-1 transcripts. The attenuation in gag mRNA translation is exacerbated by K51A substitution in the Tat double-stranded RNA-binding domain. Tat, plant virus RSS, or Dicer downregulation rescues robust gag translation and bolsters HIV-1 virion production. The reversal of HIV-1 translation repression by plant RSS supports the recent finding in Arabidopsis that plant miRNAs operate by translational inhibition. Our results identify common features between RNA silencing suppression of plant and animal viruses. We suggest that RNA silencing-mediated translation repression plays a strategic role in determining the viral set-point in a newly HIV-1-infected patient. PMID- 19122142 TI - Inferring biological mechanisms from spatial analysis: prediction of a local inhibitor in the ovary. AB - Female mammals are born with a lifetime's supply of oocytes individually enveloped in flattened epithelial cells to form primordial follicles. It is not clear how sufficient primordial follicles are maintained to sustain the reproductive lifespan, while providing an adequate supply of mature oocytes for ovulation. Locally produced growth factors are thought to be critical regulators of early follicle growth, but knowledge of their identity and source remains incomplete. Here, we have used a simple approach of spatial analysis of structures in histological tissue sections to identify likely sources of such regulatory molecules, narrowing the field for future screening for candidate growth factors or antagonists. We have quantified the relative spatial positions of primordial (resting) follicles and growing follicles in mice on days 4, 8, and 12 after birth, and calculated interfollicular distances. Follicles were significantly less likely to have started growing if they had 1 or more primordial follicles close by (within 10 mum), predicting that primordial follicles inhibit each other. This approach allows us to hypothesize that primordial follicles produce a diffusible inhibitor that prevents neighboring primordial follicles from growing. Such an approach has wide applicability within many branches of developmental and cell biology for studying spatial signaling within tissues and cells. PMID- 19122143 TI - Quantifying the global cellular thiol-disulfide status. AB - It is widely accepted that the redox status of protein thiols is of central importance to protein structure and folding and that glutathione is an important low-molecular-mass redox regulator. However, the total cellular pools of thiols and disulfides and their relative abundance have never been determined. In this study, we have assembled a global picture of the cellular thiol-disulfide status in cultured mammalian cells. We have quantified the absolute levels of protein thiols, protein disulfides, and glutathionylated protein (PSSG) in all cellular protein, including membrane proteins. These data were combined with quantification of reduced and oxidized glutathione in the same cells. Of the total protein cysteines, 6% and 9.6% are engaged in disulfide bond formation in HEK and HeLa cells, respectively. Furthermore, the steady-state level of PSSG is <0.1% of the total protein cysteines in both cell types. However, when cells are exposed to a sublethal dose of the thiol-specific oxidant diamide, PSSG levels increase to >15% of all protein cysteine. Glutathione is typically characterized as the "cellular redox buffer"; nevertheless, our data show that protein thiols represent a larger active redox pool than glutathione. Accordingly, protein thiols are likely to be directly involved in the cellular defense against oxidative stress. PMID- 19122144 TI - HER2YVMA drives rapid development of adenosquamous lung tumors in mice that are sensitive to BIBW2992 and rapamycin combination therapy. AB - Mutations in the HER2 kinase domain have been identified in human clinical lung cancer specimens. Here we demonstrate that inducible expression of the most common HER2 mutant (HER2(YVMA)) in mouse lung epithelium causes invasive adenosquamous carcinomas restricted to proximal and distal bronchioles. Continuous expression of HER2(YVMA) is essential for tumor maintenance, suggesting a key role for HER2 in lung adenosquamous tumorigenesis. Preclinical studies assessing the in vivo effect of erlotinib, trastuzumab, BIBW2992, and/or rapamycin on HER2(YVMA) transgenic mice or H1781 xenografts with documented tumor burden revealed that the combination of BIBW2992 and rapamycin is the most effective treatment paradigm causing significant tumor shrinkage. Immunohistochemical analysis of lung tumors treated with BIBW2992 and rapamycin combination revealed decreased phosphorylation levels for proteins in both upstream and downstream arms of MAPK and Akt/mTOR signaling axes, indicating inhibition of these pathways. Based on these findings, clinical testing of the BIBW2992/rapamycin combination in non-small cell lung cancer patients with tumors expressing HER2 mutations is warranted. PMID- 19122145 TI - Direct imaging of human Rad51 nucleoprotein dynamics on individual DNA molecules. AB - Rad51 protein (Rad51) is central to recombinational repair of double-strand DNA breaks. It polymerizes onto DNA and promotes strand exchange between homologous chromosomes. We visualized the real-time assembly and disassembly of human Rad51 nucleoprotein filaments on double-stranded DNA by single-molecule fluorescence microscopy. Rad51 assembly extends the DNA by approximately 65%. Nucleoprotein filament formation occurs via rapid nucleation followed by growth from these nuclei. Growth does not continue indefinitely, however, and nucleoprotein filaments terminate when approximately 2 mum in length. The dependence of nascent filament formation on Rad51 concentration suggests that 2-3 Rad51 monomers are involved in nucleation. Rad51 nucleoprotein filaments are stable and remain extended when ATP hydrolysis is prevented; however, when permitted, filaments decrease in length as a result of conversion to ADP-bound nucleoprotein complexes and partial protein dissociation. Dissociation of Rad51 from dsDNA is slow and incomplete, thereby rationalizing the need for other proteins that facilitate disassembly. PMID- 19122146 TI - HLA class I molecules consistently present internal influenza epitopes. AB - Cytotoxic T lymphocytes (CTL) limit influenza virus replication and prevent morbidity and mortality upon recognition of HLA class I presented epitopes on the surface of virus infected cells, yet the number and origin of the viral epitopes that decorate the infected cell are unknown. To understand the presentation of influenza virus ligands by human MHC class I molecules, HLA-B*0702-presented viral peptides were directly identified following influenza infection. After transfection with soluble class I molecules, peptide ligands unique to infected cells were eluted from isolated MHC molecules and identified by comparative mass spectrometry (MS). Then CTL were gathered following infection with influenza and viral peptides were tested for immune recognition. We found that the class I molecule B*0702 presents 3-6 viral ligands following infection with different strains of influenza. Peptide ligands derived from the internal viral nucleoprotein (NP(418-426) and NP(473-481)) and from the internal viral polymerase subunit PB1 (PB1(329-337)) were presented by B*0702 following infection with each of 3 different influenza strains; ligands NP(418-426), NP(473 481), and PB1(329-337) derived from internal viral proteins were consistently revealed by class I HLA. In contrast, ligands derived from hemagglutinin (HA) and matrix protein (M1) were presented intermittently on a strain-by-strain basis. When tested for immune recognition, HLA-B*0702 transgenic mice responded to NP(418-426) and PB1(329-337) consistently and NP(473-481) intermittently while ligands from HA and M1 were not recognized. These data demonstrate an emerging pattern whereby class I HLA reveal a handful of internal viral ligands and whereby CTL recognize consistently presented influenza ligands. PMID- 19122147 TI - Crystal structure of the conserved N-terminal domain of the peroxisomal matrix protein import receptor, Pex14p. AB - Pex14p is a central component of the peroxisomal protein import machinery, in which the conserved N-terminal domain mediates dynamic interactions with other peroxins including Pex5p, Pex13p, and Pex19p. Here, we report the crystal structure of the conserved N-terminal domain of Pex14p with a three-helix bundle. A hydrophobic surface is composed of the conserved residues, of which two phenylalanine residues (Phe-35 and Phe-52) protrude to the solvent. Consequently, two putative binding pockets suitable for recognizing the helical WXXXF/Y motif of Pex5p are formed on the surface by the two phenylalanine residues accompanying with positively charged residues. The structural feature agrees well with our earlier findings where F35A/L36A and F52A/L53A mutants were impaired in the interactions with other peroxins such as Pex5p and Pex13p. Pex14p variants each with Phe-to-Ala mutation at positions 35, 52, and 35/52, respectively, were defective in restoring the impaired peroxisomal protein import in pex14 Chinese hamster ovary mutant ZP161 cells. Moreover, in GST pull-down assays His(6)-Pex5pL bound only to GST-Pex14p(25-70), not to any of GST-Pex14p(25-70)F35A, GST Pex14p(25-70)F52A, and GST-Pex14p(25-70)F35A/F52A. Endogenous Pex5p was recruited to FLAG-Pex14p on peroxisomes in vivo but barely to FLAG-Pex14pF35A, FLAG Pex14pF52A, and FLAG-Pex14pF35A/F52A. Collectively, Phe-35 and Phe-52 are essential for the Pex14p functions, including the interaction between Pex14p and Pex5p. PMID- 19122148 TI - Increasing Cu bioavailability inhibits Abeta oligomers and tau phosphorylation. AB - Cognitive decline in Alzheimer's disease (AD) involves pathological accumulation of synaptotoxic amyloid-beta (Abeta) oligomers and hyperphosphorylated tau. Because recent evidence indicates that glycogen synthase kinase 3beta (GSK3beta) activity regulates these neurotoxic pathways, we developed an AD therapeutic strategy to target GSK3beta. The strategy involves the use of copper bis(thiosemicarbazonoto) complexes to increase intracellular copper bioavailability and inhibit GSK3beta through activation of an Akt signaling pathway. Our lead compound Cu(II)(gtsm) significantly inhibited GSK3beta in the brains of APP/PS1 transgenic AD model mice. Cu(II)(gtsm) also decreased the abundance of Abeta trimers and phosphorylated tau, and restored performance of AD mice in the Y-maze test to levels expected for cognitively normal animals. Improvement in the Y-maze correlated directly with decreased Abeta trimer levels. This study demonstrates that increasing intracellular copper bioavailability can restore cognitive function by inhibiting the accumulation of neurotoxic Abeta trimers and phosphorylated tau. PMID- 19122149 TI - Stromal growth and epithelial cell proliferation in ventral prostates of liver X receptor knockout mice. AB - With specific liver X receptor alpha and beta (LXRalpha and LXRbeta) antibodies, we found that LXRalpha is strongly expressed in the luminal and basal cells of prostatic epithelium. The ventral prostates (VP) of LXRalpha(-/-) mice are characterized by the presence of smooth-muscle actin-positive stromal overgrowth around the prostatic ducts and by numerous fibrous nodules pushing into the ducts and causing obstruction, so that most of the ducts were extremely dilated. BrdU labeling and Ki67 staining revealed epithelial and stromal proliferation in the fibrous nodules. However, the dense stroma surrounding the ducts was not positive for proliferation markers. There was no detectable difference between WT and LXRalpha(-/-) mice VP in the expression of the androgen receptor, but there was an increase in nuclear expression of Snail and Smad 2/3, indicating enhanced TGF beta signaling. Upon treatment of WT mice for 3 months with the LXR agonist T2320 or for 3 weeks with beta-sitosterol, LXRalpha was downregulated, and a VP phenotype similar to that of LXRalpha(-/-) mice resulted. We conclude that in rodents, LXRalpha seems to control VP stromal growth and that LXRalpha(-/-) mice may be a useful model to study prostatic stromal hyperplasia. Because LXRalpha is expressed in the epithelium, the excessive stromal growth in LXRalpha(-/-) mice indicates that LXRalpha is essential for epithelial stromal communication. PMID- 19122151 TI - Structure of the Ebola VP35 interferon inhibitory domain. AB - Ebola viruses (EBOVs) cause rare but highly fatal outbreaks of viral hemorrhagic fever in humans, and approved treatments for these infections are currently lacking. The Ebola VP35 protein is multifunctional, acting as a component of the viral RNA polymerase complex, a viral assembly factor, and an inhibitor of host interferon (IFN) production. Mutation of select basic residues within the C terminal half of VP35 abrogates its dsRNA-binding activity, impairs VP35-mediated IFN antagonism, and attenuates EBOV growth in vitro and in vivo. Because VP35 contributes to viral escape from host innate immunity and is required for EBOV virulence, understanding the structural basis for VP35 dsRNA binding, which correlates with suppression of IFN activity, is of high importance. Here, we report the structure of the C-terminal VP35 IFN inhibitory domain (IID) solved to a resolution of 1.4 A and show that VP35 IID forms a unique fold. In the structure, we identify 2 basic residue clusters, one of which is important for dsRNA binding. The dsRNA binding cluster is centered on Arg-312, a highly conserved residue required for IFN inhibition. Mutation of residues within this cluster significantly changes the surface electrostatic potential and diminishes dsRNA binding activity. The high-resolution structure and the identification of the conserved dsRNA binding residue cluster provide opportunities for antiviral therapeutic design. Our results suggest a structure-based model for dsRNA mediated innate immune antagonism by Ebola VP35 and other similarly constructed viral antagonists. PMID- 19122150 TI - Ribosome-induced changes in elongation factor Tu conformation control GTP hydrolysis. AB - In translation, elongation factor Tu (EF-Tu) molecules deliver aminoacyl-tRNAs to the mRNA-programmed ribosome. The GTPase activity of EF-Tu is triggered by ribosome-induced conformational changes of the factor that play a pivotal role in the selection of the cognate aminoacyl-tRNAs. We present a 6.7-A cryo-electron microscopy map of the aminoacyl-tRNA x EF-Tu x GDP x kirromycin-bound Escherichia coli ribosome, together with an atomic model of the complex obtained through molecular dynamics flexible fitting. The model reveals the conformational changes in the conserved GTPase switch regions of EF-Tu that trigger hydrolysis of GTP, along with key interactions, including those between the sarcin-ricin loop and the P loop of EF-Tu, and between the effector loop of EF-Tu and a conserved region of the 16S rRNA. Our data suggest that GTP hydrolysis on EF-Tu is controlled through a hydrophobic gate mechanism. PMID- 19122152 TI - Interactive Cardio-Vascular and Thoracic Surgery virtual conferences are now printed monthly: more at www.icvts.org. PMID- 19122153 TI - eComment: Tracheal surgery. PMID- 19122154 TI - eComment: Early extubation following tracheal surgery. Is it safe? PMID- 19122155 TI - eComment: Freedom Solo stentless aortic bioprosthesis and postoperative thrombocytopenia--interpretation of available data and clinical consequences for surgeons. PMID- 19122156 TI - eComment: Lower postoperative platelet levels after aortic valve replacement with Freedom Solo prostheses: are there clinical repercussions? PMID- 19122157 TI - eComment: Long-term outcomes in octogenarians with lung cancer following mediastinal lymph node dissection. PMID- 19122158 TI - eComment: Re: The impact of allogenic red cell transfusion and coated bypass circuit on the inflammatory response during cardiopulmonary bypass: a randomized study. PMID- 19122159 TI - eComment: An infected pseudoaneurysm caused by methicillin-resistant Staphylococcus aureus. PMID- 19122160 TI - eComment: Surgical treatment of coarctation in adult patients yields better long term results with regard to hypertension but carries a substantial risk. PMID- 19122161 TI - eComment: Questioning a 'best evidence' search arguing against abdominal-only cardiopulmonary resuscitation and for external cardiac massage. PMID- 19122162 TI - eComment: Everyday clinical practice consequences? PMID- 19122163 TI - eComment: Unexpected chylopericardium and its treatment after cardiothoracic operations. PMID- 19122164 TI - Atrial natriuretic peptide increases inflammation, infarct size, and mortality after experimental coronary occlusion. AB - Acute coronary artery occlusion triggers the release of atrial natriuretic peptide (ANP) from the heart. ANP affects vasodilation, natriuresis, and inflammation, but the integrated biological effects of ANP on myocardial infarction are unknown. To elucidate these effects, the left anterior coronary artery was ligated in anesthetized, ANP-deficient (ANP(-/-)) and congenic wild type (ANP(+/+)) mice. The survival of ANP(-/-) mice was markedly better (56%) at 30 days postinfarction than the survival of ANP(+/+) mice (20%, P < 0.01). Surviving mice were comparable initially, but ANP(-/-) mice developed more cardiac hypertrophy (P < 0.001) and had lower contractility indexes 30 days after infarction (P < 0.05). An analysis 24 h after coronary occlusion showed that ANP( /-) mice had smaller infarcts than ANP(+/+) mice (62.6 +/- 12.1 vs. 100.8 +/- 3.8%, P < 0.001) adjusted for comparable areas at risk for ischemia. The administration of ANP to ANP(-/-) mice via osmotic minipumps significantly enlarged infarct size to levels comparable with those observed in ANP(+/+) mice (P < 0.05). There was no difference in neutrophil migration into the noninfarcted myocardium of ANP(-/-) mice undergoing actual versus sham-operated coronary occlusion. By comparison, after coronary occlusion, the neutrophil infiltration into the myocardium was enhanced in ANP(+/+) (P < 0.0005) and ANP(-/-) mice administered ANP (P < 0.0005). The expression of P-selectin, a molecule that mediates neutrophil adhesion, was significantly greater after coronary occlusion in the vasculature of ANP(+/+) or ANP(-/-) mice treated with ANP than in ANP(-/-) mice (P < 0.002). Taken together, these results indicate that ANP increases P selectin, neutrophil infiltration, infarct size, and mortality following experimental coronary occlusion. PMID- 19122165 TI - Phosphoinositide 3-kinase Akt signaling pathway interacts with protein kinase Cbeta2 in the regulation of physiologic developmental hypertrophy and heart function. AB - The phosphoinositide 3-kinase (PI3-kinase)-protein kinase B (Akt) signaling pathway is essential in the induction of physiological cardiac hypertrophy. In contrast, protein kinase C beta2 (PKCbeta2) is implicated in the development of pathological cardiac hypertrophy and heart failure. Thus far, no clear association has been demonstrated between these two pathways. In this study, we examined the potential interaction between the PI3-kinase and PKCbeta2 pathways by crossing transgenic mice with cardiac specific expression of PKCbeta2, constitutively active (ca) PI3-kinase, and dominant-negative (dn) PI3-kinase. In caPI3-kinase/PKCbeta2 and dnPI3-kinase/PKCbeta2 double-transgenic mice, the heart weight-to-body weight ratios and cardiomyocyte sizes were similar to those observed in caPI3-kinase and dnPI3-kinase transgenic mice, respectively, suggesting that the regulation of physiological developmental hypertrophy via modulation of cardiomyocyte size proceeds through the PI3-kinase pathway. In addition, we observed that caPI3-kinase/PKCbeta2 mice showed improved cardiac function while the function of dnPI3-kinase/PKCbeta2 mice was similar to that of the PKCbeta2 group. PKCbeta2 protein levels in both dnPI3-kinase/PKCbeta2 and PKCbeta2 mice were significantly upregulated. Interestingly, however, PKCbeta2 protein expression was significantly attenuated in caPI3-kinase/PKCbeta2 mice. PI3-kinase activity measured by Akt phosphorylation was not affected by PKCbeta2 overexpression. These data suggest a potential interaction between these two pathways in the heart, where PI3-kinase is predominantly responsible for the regulation of physiological developmental hypertrophy and may act as an upstream modulator of PKCbeta2 with the potential for rescuing the pathological cardiac dysfunction induced by overexpression of PKCbeta. PMID- 19122166 TI - Left ventricular regional wall curvedness and wall stress in patients with ischemic dilated cardiomyopathy. AB - Geometric remodeling of the left ventricle (LV) after myocardial infarction is associated with changes in myocardial wall stress. The objective of this study was to determine the regional curvatures and wall stress based on three dimensional (3-D) reconstructions of the LV using MRI. Ten patients with ischemic dilated cardiomyopathy (IDCM) and 10 normal subjects underwent MRI scan. The IDCM patients also underwent delayed gadolinium-enhancement imaging to delineate the extent of myocardial infarct. Regional curvedness, local radii of curvature, and wall thickness were calculated. The percent curvedness change between end diastole and end systole was also calculated. In normal heart, a short- and long axis two-dimensional analysis showed a 41 +/- 11% and 45 +/- 12% increase of the mean of peak systolic wall stress between basal and apical sections, respectively. However, 3-D analysis showed no significant difference in peak systolic wall stress from basal and apical sections (P = 0.298, ANOVA). LV shape differed between IDCM patients and normal subjects in several ways: LV shape was more spherical (sphericity index = 0.62 +/- 0.08 vs. 0.52 +/- 0.06, P < 0.05), curvedness at end diastole (mean for 16 segments = 0.034 +/- 0.0056 vs. 0.040 +/- 0.0071 mm(-1), P < 0.001) and end systole (mean for 16 segments = 0.037 +/- 0.0068 vs. 0.067 +/- 0.020 mm(-1), P < 0.001) was affected by infarction, and peak systolic wall stress was significantly increased at each segment in IDCM patients. The 3-D quantification of regional wall stress by cardiac MRI provides more precise evaluation of cardiac mechanics. Identification of regional curvedness and wall stresses helps delineate the mechanisms of LV remodeling in IDCM and may help guide therapeutic LV restoration. PMID- 19122167 TI - Lymphatic pump-conduit duality: contraction of postnodal lymphatic vessels inhibits passive flow. AB - Lymphangions, the segments of lymphatic vessels between valves, exhibit structural characteristics in common with both ventricles and arteries. Although once viewed as passive conduits like arteries, it has become well established that lymphangions can actively pump lymph against an axial pressure gradient from low-pressure tissues to the great veins of the neck. A recently reported mathematical model, based on fundamental principles, predicted that lymphangions can transition from pump to conduit behavior when outlet pressure falls below inlet pressure. In this case, the axial pressure gradient becomes the major source of energy for the propulsion of lymph, despite the presence of cyclical contraction. In fact, flow is augmented when cyclical contractions are abolished. We therefore used an in vitro preparation to confirm these findings and to test the hypothesis that lymphangion contraction inhibits flow when outlet pressure falls below inlet pressure. Bovine postnodal mesenteric lymphatic vessels harvested from an abattoir were subjected to an inlet pressure of 5.0 cmH(2)O and an outlet pressure that decreased from 6.5 to 3.5 cmH(2)O under control conditions, stimulated with U-46619 (a thromboxane analog) and relaxed with calcium-free solution. Under control conditions, lymphatic flow markedly increased as outlet pressure fell below inlet pressure. In this case, the slopes of the flow versus axial pressure gradient increased with calcium-free conditions (61%, n = 8, P = 0.016) and decreased with U-46619 stimulation (21%, n = 5, P = 0.033). Our findings indicate that the stimulation of lymphatic contractility does indeed inhibit lymphatic flow when vessels act like conduits. PMID- 19122168 TI - A novel echocardiographic marker of end systole in the ischemic left ventricle: "tug of war" sign. AB - The present study introduces a new clinical method to define left ventricular (LV) end systole (ES) during tissue Doppler imaging (TDI). Preliminary experiments showed a sharp inflection point in strain traces (S(IP)) from ischemic borderzones, which coincided with onset of a postsystolic shortening wave (V(PS)) in the velocity trace. In a single-vessel disease model, we investigated whether S(IP) and V(PS) may serve as markers of global ES and their mechanism. In six anesthetized dogs we measured LV pressure and myocardial long axis function by using TDI and sonomicrometry. Ischemia was induced by left anterior descending coronary artery occlusion. ES was defined by the minimum first derivative of LV pressure. TDI and sonomicrometry demonstrated a sharp S(IP) and V(PS) at ES in the ischemic borderzone (defined as moderately ischemic myocardium by pressure-dimension loop analysis). Time differences relative to ES ( +/- SD) were -0.1 +/- 2.3 (intraclass correlation coefficient R(IC) = 0.996) and 6.8 +/- 10.7 ms (R(IC) = 0.89) for S(IP) as shown by sonomicrometry and TDI, respectively. There was a strong inverse relationship between postsystolic shortening in the borderzone and simultaneous lengthening of nonischemic myocardium. In 30 patients with acute myocardial infarction, S(IP) and V(PS) evaluated by TDI were compared with ES defined by aortic valve closure. Time differences were -4 +/- 14 (R(IC) = 0.94) and -2 +/- 11 ms (R(IC) = 0.96), respectively. In the ischemic borderzone, S(IP) and V(PS) identified global ES with high accuracy. The force balance or "tug of war" between borderzone and nonischemic myocardium is a likely underlying mechanism for these markers. The method may be used as an "all in one heart beat" approach for TDI analysis in acute myocardial ischemia. PMID- 19122169 TI - Caspase-8 is involved in neovascularization-promoting progenitor cell functions. AB - OBJECTIVE: Endothelial progenitor cells (EPCs) comprise a heterogeneous population of cells, which improve therapeutic neovascularization after ischemia. The neovascularization-promoting potential of progenitor cells depends on survival and retention of the infused cells to the tissue. Caspases mediate apoptosis but are also involved in other critical biological processes. Therefore, we aimed to address the role of caspases in proangiogenic cells. METHODS AND RESULTS: The caspase-8 inhibitor zIETD abrogated the ex vivo formation of EPCs, inhibited EPC adhesion and migration, and reduced their capacity to improve neovascularization in vivo. Consistently, cells isolated from caspase-8-deficient mice exhibited a reduced capacity for enhancing neovascularization when transplanted into mice after hindlimb ischemia. Because inhibition of Caspase-8 reduced the adhesion and homing functions of EPCs, we further determined the surface expression of integrins and receptors involved in cell recruitment to ischemic tissues. Pharmacological inhibition of caspase-8 and genetic depletion of caspase-8 reduced the expression of the fibronectin receptor subunits alpha5 and beta1 and the SDF-1 receptor CXCR4. Moreover, we identified the E3 ubiquitin ligase Cbl-b, which negatively regulates integrin and receptor mediated signaling, as a potential Caspase-8 substrate. CONCLUSIONS: In summary, our data demonstrate a novel apoptosis-unrelated role of caspase-8 in proangiogenic cells. PMID- 19122170 TI - Prognostic utility of apoB/AI, total cholesterol/HDL, non-HDL cholesterol, or hs CRP as predictors of clinical risk in patients receiving statin therapy after acute coronary syndromes: results from PROVE IT-TIMI 22. AB - OBJECTIVES: The purpose of this study was to compare the prognostic utility of apoB/AI, total cholesterol/HDL (TC/HDL) ratio, non-HDL cholesterol (non-HDL-C), or hs-CRP as predictors of clinical risk among patients receiving statin therapy after acute coronary syndromes (ACS). METHODS AND RESULTS: Patients with ACS were randomized in the PROVE IT-TIMI 22 trial to either pravastatin 40 mg or atorvastatin 80 mg. Cox regression models adjusting for confounders were used to assess the relationship between on-treatment lipids or hs-CRP and risk of death or acute coronary events. At 4 months a 1 SD increment in apoB/AI (HR 1.10, 95% CI 1.01 to 1.20), TC/HDL (HR 1.12, 95% CI 1.01 to 1.24), and non-HDL-C (HR 1.20, 95% CI 1.07 to 1.35) predicted events to a similar extent as LDL-C (HR 1.20, 95% CI 1.07 to 1.35) with neither apoB/AI, TC/HDL, nor non-HDL-C improving risk prediction models which included LDL-C. In contrast, the addition of hs-CRP significantly improved risk prediction models irrespective of the lipid parameters included, with a 29% to 30% increased risk observed per 1 SD increment in log CRP. CONCLUSION: In the present study of ACS patients receiving statin therapy, on-treatment apoB/AI, TC/HDL, and non-HDL-C offered similar prognostic information to LDL-C. However, the addition of hs-CRP to lipid-based measurements significantly improved risk prediction. On treatment CRP measurement may therefore offer additive prognostic information to lipids in ACS patients. PMID- 19122171 TI - Insulin-like growth factor-1 receptor expression masks the antiinflammatory and glucose uptake capacity of insulin in vascular smooth muscle cells. AB - OBJECTIVE: Insulin resistance of vascular smooth muscle cells (VSMCs) has been linked to accelerated atherosclerosis in diabetes; however, the effects of insulin on VSMCs remain controversial. Most VSMC insulin receptors are sequestered into insulin-insensitive hybrids with insulin-like growth factor-1 receptors (IGF1Rs). Thus we hypothesized that regulation of IGF1R expression may impact cellular insulin sensitivity. METHODS AND RESULTS: IGF1R expression was increased in aortas from diabetic mice. IGF1R overexpression in VSMCs impaired insulin-induced Akt phosphorylation. Conversely, IGF1R downregulation by siRNA allowed assembly of insulin holoreceptors, enhanced insulin-induced phosphorylation of its receptor, Akt, Erk1/2, and further augmented insulin induced glucose uptake. IGF1R downregulation uncovered an insulin-induced reduction in activation of NF-kappaB and inhibition of MCP-1 upregulation in response to TNF-alpha. CONCLUSIONS: Downregulation of IGF1R increases the fraction of insulin receptors organized in holoreceptors, which leads to enhanced insulin signaling and unmasks potential antiinflammatory properties of insulin in VSMCs. Therefore, IGF1R, which is susceptible to feedback regulation by its own ligand, may represent a novel target for interventions designed to treat insulin resistance in the vasculature. PMID- 19122172 TI - Fibrinogen-gamma C-terminal fragments induce endothelial barrier dysfunction and microvascular leak via integrin-mediated and RhoA-dependent mechanism. AB - OBJECTIVES: The purposes of this study were to characterize the direct effect of the C-terminal fragment of fibrinogen gamma chain (gammaC) on microvascular endothelial permeability and to examine its molecular mechanism of action. METHODS AND RESULTS: Intravital microscopy was performed to measure albumin extravasation in intact mesenteric microvasculature, followed by quantification of hydraulic conductivity in single perfused microvessels. Transendothelial electric resistance was measured in microvascular endothelial cells in combination with immunoblotting and immunocytochemistry. The results show that gammaC induced time- and concentration-dependent increases in protein transvascular flux and water permeability and decreases in endothelial barrier function, coupled with Rho GTPase activation, myosin light chain phosphorylation, and stress fiber formation. Depletion of RhoA via siRNA knockdown or pharmacological inhibition of RhoA signaling attenuated gammaC-induced barrier dysfunction. Imaging analyses demonstrated binding of gammaC to endothelial cells; the interaction was inhibited during blockage of the alphavbeta3 integrin. Furthermore, in vivo experiments showed that the microvascular leak response to gammaC was attenuated in integrin beta3(-/-) animals. CONCLUSION: Fibrinogen gamma C terminus directly interacts with the microvascular endothelium causing fluid and protein leak. The endothelial response to gammaC involves an integrin receptor-mediated RhoA-dependent signaling pathway that leads to paracellular hyperpermeability. PMID- 19122173 TI - Statins control oxidized LDL-mediated histone modifications and gene expression in cultured human endothelial cells. AB - OBJECTIVE: Activation of the endothelium by oxidized low-density lipoprotein (oxLDL) has been implicated in the development of atherosclerosis. Histone modifications impact on the transcriptional activity state of genes. We tested the hypothesis that oxLDL-induced inflammatory gene expression is regulated by histone modifications and experienced the effect of statins on these alterations. METHODS AND RESULTS: OxLDL-related interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) secretion in endothelial cells was reduced by statins but enhanced by histone deacetylase inhibitors. OxLDL induced lectin-like oxidized LDL receptor-1 (LOX-1) and extracellular regulated kinases (ERK1/2) dependent acetylation of histone H3 and H4 as well as phosphorylation of histone H3, both globally and on the promoters of il8 and mcp1. Pretreatment of oxLDL exposed cells with statins reduced the above mentioned histone modification, as well as recruitment of CREB binding protein (CBP) 300, NF-kappaB, and of RNA polymerase II but prevented loss of binding of histone deacetylase (HDAC)-1 and 2 at the il8 and mcp1 gene promoters. OxLDL reduced HDAC1 and 2 expression, and statins partly restored global HDAC-activity. Statin-related effects were reverted with mevalonate. In situ experiments indicated decreased expression of HDAC2 in endothelial cells in atherosclerotic plaques of human coronary arteries. CONCLUSIONS: Histone modifications seem to play an important role in atherosclerosis. PMID- 19122174 TI - Genetic regulation of atherosclerotic plaque size and morphology in the innominate artery of hyperlipidemic mice. AB - OBJECTIVE: We sought to determine the genetic factors contributing to atherosclerotic plaque size and cellular composition in the innominate artery, a murine model of advanced atherosclerosis. METHODS AND RESULTS: We examined genetic contributions to innominate atherosclerotic plaque size and cellular composition in an intercross between C57BL/6J.Apoe(-/-), a strain susceptible to aortic lesions, and C3H/HeJ.Apoe(-/-), a strain resistant to aortic lesions. Surprisingly, total innominate lesion size was similar in the two strains. Genetic analyses identified one novel locus on Chromosome 2 for innominate artery lesion size, a significant locus for fibrous cap thickness on Chromosome 15, and several suggestive loci for cellular composition, all distinct from loci influencing aortic lesions. The Chromosome 2 locus contains a candidate, CD44. We show that CD44 is expressed in the innominate artery and differs strikingly in expression between the parental strains. CONCLUSION: Multiple aspects of innominate lesion composition are genetically determined, but in a manner largely independent of the genetic contributions to aortic lesions. PMID- 19122175 TI - Induction of prostacyclin by steady laminar shear stress suppresses tumor necrosis factor-alpha biosynthesis via heme oxygenase-1 in human endothelial cells. AB - Cyclooxygenase (COX)-2 is among the endothelial genes upregulated by uniform laminar shear stress (LSS), characteristically associated with atherosclerotic lesion-protected areas. We have addressed whether the induction of COX-2 dependent prostanoids in endothelial cells by LSS plays a role in restraining endothelial tumor necrosis factor (TNF)-alpha generation, a proatherogenic cytokine, through the induction of heme oxygenase-1 (HO)-1, an antioxidant enzyme. In human umbilical vein endothelial cells (HUVECs) exposed to steady LSS of 10 dyn/cm(2) for 6 hours, COX-2 protein was significantly induced, whereas COX 1 and the downstream synthases were not significantly modulated. This was associated with significant (P<0.05) increase of 6-keto-prostaglandin (PG)F(1alpha) (the hydrolysis product of prostacyclin), PGE(2), and PGD(2). In contrast, TNF-alpha released in the medium in 6 hours (3633+/-882 pg) or detected in cells lysates (1091+/-270 pg) was significantly (P<0.05) reduced versus static condition (9100+/-2158 and 2208+/-300 pg, respectively). Coincident induction of HO-1 was detected. The finding that LSS-dependent reduction of TNF-alpha generation and HO-1 induction were abrogated by the selective inhibitor of COX-2 NS-398, the nonselective COX inhibitor aspirin, or the specific prostacyclin receptor (IP) antagonist RO3244794 illuminates the central role played by LSS induced COX-2-dependent prostacyclin in restraining endothelial inflammation. Carbacyclin, an agonist of IP, induced HO-1. Similarly to inhibition of prostacyclin biosynthesis or activity, the novel imidazole-based HO-1 inhibitor QC15 reversed TNF-alpha reduction by LSS. These findings suggest that inhibition of COX-2-dependent prostacyclin might contribute to acceleration of atherogenesis in patients taking traditional nonsteroidal antiinflammatory drugs (NSAIDs) and NSAIDs selective for COX-2 through downregulation of HO-1, which halts TNF-alpha generation in human endothelial cells. PMID- 19122176 TI - The antiangiogenic activity of rPAI-1(23) inhibits vasa vasorum and growth of atherosclerotic plaque. AB - Plaque vascularity has been implicated in its growth and stability. However, there is a paucity of information regarding the origin of plaque vasculature and the role of vasa vasorum in plaque growth. To inhibit growth of vasa vasorum in atherogenic mice and assess its effect on plaque growth, we used a truncated plasminogen activator inhibitor (PAI)-1 protein, rPAI-1(23), that has significant antiangiogenic activity. Female LDLR(-/-)ApoB-48-deficient mice fed Paigen's diet without cholate for 20 weeks received rPAI-1(23) treatment (n=21) for the last 6 weeks. Plaque size and vasa vasorum density were compared to 2 controls: mice fed Paigen's diet and treated with saline for the last 6 weeks (n=16) and mice fed Paigen's diet until the onset of treatment (n=14). The rPAI-1(23) treatment significantly reduced plaque area and plaque cholesterol in the descending aorta and plaque area in the innominate artery. Measurements of reconstructed confocal microscopy images of vasa vasorum demonstrate that rPAI-1(23) treatment decreased vasa vasorum area and length, which was supported by microCT images. Confocal images provide evidence for vascularized plaque in the saline-treated group but not in rPAI-1(23)-treated mice. The increased vessel density in saline-treated mice is attributable, in part, to upregulated fibroblast growth factor-2 expression, which is inhibited by rPAI-1(23). In conclusion, rPAI-1(23) inhibits growth of vasa vasorum, as well as vessels within the adjacent plaque and vessel wall, through inhibition of fibroblast growth factor-2, leading to reduced plaque growth in atherogenic female LDLR(-/-)ApoB-48-deficient mice. PMID- 19122177 TI - The Ca(v)3.2 T-type Ca(2+) channel is required for pressure overload-induced cardiac hypertrophy in mice. AB - Voltage-gated T-type Ca(2+) channels (T-channels) are normally expressed during embryonic development in ventricular myocytes but are undetectable in adult ventricular myocytes. Interestingly, T-channels are reexpressed in hypertrophied or failing hearts. It is unclear whether T-channels play a role in the pathogenesis of cardiomyopathy and what the mechanism might be. Here we show that the alpha(1H) voltage-gated T-type Ca(2+) channel (Ca(v)3.2) is involved in the pathogenesis of cardiac hypertrophy via the activation of calcineurin/nuclear factor of activated T cells (NFAT) pathway. Specifically, pressure overload induced hypertrophy was severely suppressed in mice deficient for Ca(v)3.2 (Ca(v)3.2(-/-)) but not in mice deficient for Ca(v)3.1 (Ca(v)3.1(-/-)). Angiotensin II-induced cardiac hypertrophy was also suppressed in Ca(v)3.2(-/-) mice. Consistent with these findings, cultured neonatal myocytes isolated from Ca(v)3.2(-/-) mice fail to respond hypertrophic stimulation by treatment with angiotensin II. Together, these results demonstrate the importance of Ca(v)3.2 in the development of cardiac hypertrophy both in vitro and in vivo. To test whether Ca(v)3.2 mediates the hypertrophic response through the calcineurin/NFAT pathway, we generated Ca(v)3.2(-/-), NFAT-luciferase reporter mice and showed that NFAT luciferase reporter activity failed to increase after pressure overload in the Ca(v)3.2(-/-)/NFAT-Luc mice. Our results provide strong genetic evidence that Ca(v)3.2 indeed plays a pivotal role in the induction of calcineurin/NFAT hypertrophic signaling and is crucial for the activation of pathological cardiac hypertrophy. PMID- 19122178 TI - Proinflammatory phenotype of perivascular adipocytes: influence of high-fat feeding. AB - Adipose tissue depots originate from distinct precursor cells, are functionally diverse, and modulate disease processes in a depot-specific manner. However, the functional properties of perivascular adipocytes, and their influence on disease of the blood vessel wall, remain to be determined. We show that human coronary perivascular adipocytes exhibit a reduced state of adipocytic differentiation as compared with adipocytes derived from subcutaneous and visceral (perirenal) adipose depots. Secretion of antiinflammatory adiponectin is markedly reduced, whereas that of proinflammatory cytokines interleukin-6, interleukin-8, and monocyte chemoattractant protein-1, is markedly increased in perivascular adipocytes. These depot-specific differences in adipocyte function are demonstrable in both freshly isolated adipose tissues and in vitro-differentiated adipocytes. Murine aortic arch perivascular adipose tissues likewise express lower levels of adipocyte-associated genes as compared with subcutaneous and visceral adipose tissues. Moreover, 2 weeks of high-fat feeding caused further reductions in adipocyte-associated gene expression, while upregulating proinflammatory gene expression, in perivascular adipose tissues. These changes were observed in the absence of macrophage recruitment to the perivascular adipose depot. We conclude that perivascular adipocytes exhibit reduced differentiation and a heightened proinflammatory state, properties that are intrinsic to the adipocytes residing in this depot. Dysfunction of perivascular adipose tissue induced by fat feeding suggests that this unique adipose depot is capable of linking metabolic signals to inflammation in the blood vessel wall. PMID- 19122180 TI - IK1 heterogeneity affects genesis and stability of spiral waves in cardiac myocyte monolayers. AB - Previous studies have postulated an important role for the inwardly rectifying potassium current (I(K1)) in controlling the dynamics of electrophysiological spiral waves responsible for ventricular tachycardia and fibrillation. In this study, we developed a novel tissue model of cultured neonatal rat ventricular myocytes (NRVMs) with uniform or heterogeneous Kir2.1expression achieved by lentiviral transfer to elucidate the role of I(K1) in cardiac arrhythmogenesis. Kir2.1-overexpressed NRVMs showed increased I(K1) density, hyperpolarized resting membrane potential, and increased action potential upstroke velocity compared with green fluorescent protein-transduced NRVMs. Opposite results were observed in Kir2.1-suppressed NRVMs. Optical mapping of uniformly Kir2.1 gene-modified monolayers showed altered conduction velocity and action potential duration compared with nontransduced and empty vector-transduced monolayers, but functional reentrant waves could not be induced. In monolayers with an island of altered Kir2.1 expression, conduction velocity and action potential duration of the locally transduced and nontransduced regions were similar to those of the uniformly transduced and nontransduced monolayers, respectively, and functional reentrant waves could be induced. The waves were anchored to islands of Kir2.1 overexpression and remained stable but dropped in frequency and meandered away from islands of Kir2.1 suppression. In monolayers with an inverse pattern of I(K1) heterogeneity, stable high frequency spiral waves were present with I(K1) overexpression, whereas lower frequency, meandering spiral waves were observed with I(K1) suppression. Our study provides direct evidence for the contribution of I(K1) heterogeneity and level to the genesis and stability of spiral waves and highlights the potential importance of I(K1) as an antiarrhythmia target. PMID- 19122179 TI - Free cholesterol accumulation in macrophage membranes activates Toll-like receptors and p38 mitogen-activated protein kinase and induces cathepsin K. AB - The molecular events linking lipid accumulation in atherosclerotic plaques to complications such as aneurysm formation and plaque disruption are poorly understood. BALB/c-Apoe(-/-) mice bearing a null mutation in the Npc1 gene display prominent medial erosion and atherothrombosis, whereas their macrophages accumulate free cholesterol in late endosomes and show increased cathepsin K (Ctsk) expression. We now show increased cathepsin K immunostaining and increased cysteinyl proteinase activity using near infrared fluorescence imaging over proximal aortas of Apoe(-/-), Npc1(-/-) mice. In mechanistic studies, cholesterol loading of macrophage plasma membranes (cyclodextrin-cholesterol) or endosomal system (AcLDL+U18666A or Npc1 null mutation) activated Toll-like receptor (TLR) signaling, leading to sustained phosphorylation of p38 mitogen-activated protein kinase and induction of p38 targets, including Ctsk, S100a8, Mmp8, and Mmp14. Studies in macrophages from knockout mice showed major roles for TLR4, following plasma membrane cholesterol loading, and for TLR3, after late endosomal loading. TLR signaling via p38 led to phosphorylation and activation of the transcription factor Microphthalmia transcription factor, acting at E-box elements in the Ctsk promoter. These studies suggest that free cholesterol enrichment of either plasma or endosomal membranes in macrophages leads to activation of signaling via various TLRs, prolonged p38 mitogen-activated protein kinase activation, and induction of Mmps, Ctsk, and S100a8, potentially contributing to plaque complications. PMID- 19122182 TI - Lymphotropic polyomavirus and progressive multifocal leukoencephalopathy. PMID- 19122183 TI - Developmental programming: contribution of prenatal androgen and estrogen to estradiol feedback systems and periovulatory hormonal dynamics in sheep. AB - Prenatal testosterone excess leads to neuroendocrine and periovulatory disruptions in the offspring culminating in progressive loss of cyclicity. It is unknown whether the mediary of these disruptions is androgen or estrogen, because testosterone can be aromatized to estrogen. Taking a reproductive life span approach of studying control, prenatal testosterone, and dihydrotestosterone treated offspring, this study tested the hypothesis that disruptions in estradiol negative but not -positive feedback effects are programmed by androgenic actions of testosterone and that these disruptions in turn will have an impact on the periovulatory hormonal dynamics. The approach was to test estradiol-negative and positive feedback responses of all three groups of ovary-intact females during prepubertal age and then compare the periovulatory dynamics of luteinizing hormone, follicle-stimulating hormone, estradiol, and progesterone during the first breeding season. The findings show that estradiol-negative but not estradiol-positive feedback disruptions in prenatal testosterone-treated females are programmed by androgenic actions of prenatal testosterone excess and that follicular phase estradiol and gonadotropins surge disruptions during reproductive life are consistent with estrogenic programming. Additional studies carried out testing estradiol-positive feedback response over time found progressive deterioration of estradiol-positive feedback in prenatal testosterone treated sheep until the time of puberty. Together, these findings provide insight into the mechanisms by which prenatal testosterone disrupts the reproductive axis. The findings may be of translational relevance since daughters of mothers with hyperandrogenism are at risk of increased exposure to androgens. PMID- 19122186 TI - Involvement of polypyrimidine tract-binding protein (PTB)-related proteins in pollen germination in Arabidopsis. AB - The pollen grains of most angiosperms contain stores of RNAs and their translation products required for pollen germination and subsequent early elongation of pollen tubes. Polypyrimidine tract-binding protein (PTB), which is involved in the regulation of pre-mRNA alternative splicing, internal ribosomal entry site (IRES)-mediated translation and mRNA localization/sorting, is known to act as a bridging molecule between RNAs and a variety of cellular factors to fulfill cellular functions in both the nucleus and cytoplasm. Moreover, it has been reported that PTB plays roles in the differentiation and development of animal cells and tissues. In the Arabidopsis genome, there are two PTB-related genes, tentatively termed AtPTB1 and AtPTB2. In the present study, the physiological functions of AtPTBs were investigated using genetic and cytological approaches. The AtPTB promoter was highly active in vegetative cells of mature pollen grains, and AtPTB was localized in the nucleus and cytoplasm of these vegetative cells. Mutations in the AtPTB genes resulted in decreased germination efficiency, and this effect was rescued by introduction of the AtPTB2 promoter::AtPTB2-GFP. Taken together, these findings suggest that AtPTB is involved in pollen germination through possible RNA metabolism processes in late maturing and mature pollen grains. PMID- 19122184 TI - Juvenile rank can predict male-typical adult mating behavior in female sheep treated prenatally with testosterone. AB - Previous research with female sheep indicates that exposure to excess testosterone for 60 days (from Gestational Days 30-90 of the 147-day gestation) leads to virilized genitalia, severe neuroendocrine deficits, as well as masculinization and defeminization of sexual behavior (T60 females). In contrast, 30 days of testosterone exposure (Gestational Days 60-90) produce animals with female-typical genitalia, less severe neuroendocrine alterations, and variable gender patterns of sexual behavior (T30 females). Variation in adult sexual behavior of male ungulates is influenced by early social experience, but this has never been tested in females. Here we investigate the influence of rank in the dominance hierarchy on the expression of adult sexual behavior in females. Specifically, we hypothesized that juvenile rank would predict the amount of male and female-typical mating behavior exhibited by adult female sheep. This hypothesis was tested in two treatment groups and their controls (group 1: T60 females; group 2: T30 females). Dominance hierarchies were determined by observing competition over resources. Both groups of prenatal testosterone treated females were higher ranking than controls (T60: P = 0.05; T30: P < 0.01). During the breeding season, both T60 and T30 females exhibited more male-typical mating behavior than did controls; however, the T30 animals also exhibited female typical behavior. For the T60 group, prenatal treatment, not juvenile rank, best predicted male-typical sex behavior (P = 0.007), while juvenile rank better predicted male mating behavior for the T30 group (P = 0.006). Rank did not predict female mating behavior in the hormone-treated or control ewes. We conclude that the effect of prenatal testosterone exposure on adult male-specific but not female-specific mating behavior is modulated by juvenile social experiences. PMID- 19122187 TI - Increase in ascorbate content of transgenic tobacco plants overexpressing the acerola (Malpighia glabra) phosphomannomutase gene. AB - Phosphomannomutase (PMM; EC 5.4.2.8) catalyzes the interconversion of mannose-6 phosphate to mannose-1-phosphate in the Smirnoff-Wheeler pathway for the biosynthesis of l-ascorbic acid (AsA). We have cloned the PMM cDNA from acerola (Malpighia glabra), a plant containing an enormous amount of AsA. The AsA contents correlate with the PMM gene expression of the ripening fruits and leaves. The PMM activities in the leaves of acerola, tomato and Arabidopsis correlate with their respective AsA contents. Transgenic tobacco plants overexpressing the acerola PMM gene showed about a 2-fold increase in AsA contents compared with the wild type, with a corresponding correlation with the PMM transcript levels and activities. PMID- 19122188 TI - A novel nuclear-encoded protein, NDH-dependent cyclic electron flow 5, is essential for the accumulation of chloroplast NAD(P)H dehydrogenase complexes. AB - The chloroplast NAD(P)H dehydrogenase (NDH) complex, which reduces plastoquinones in thylakoid membranes, is involved in PSI cyclic electron flow and chlororespiration. In addition to land plants, the NDH complex is conserved in cyanobacteria. In this study, we identified a novel NDH-related gene of Arabidopsis, NDH-dependent cyclic electron flow 5 (NDF5, At1g55370). Post illumination increases in chlorophyll fluorescence were absent in ndf5 mutant plants, which indicated that NDF5 is essential for NDH activity. Sequence analysis did not reveal any known functional motifs in NDF5, but there was some homology in amino acid sequence between NDF5 and NDF2, a known NDH subunit. NDF5 and NDF2 homologs were present in higher plants, but not cyanobacteria. A single homolog, which had similarity to both NDF5 and NDF2, was identified in the moss Physcomitrella patens. Immunoblot analysis showed that NDF5 localizes to membrane fractions of chloroplasts. The stability of NdhH, a subunit of the NDH complex, as well as NDF5 and NDF2, was decreased in ndf5, ndf2 and double ndf2/ndf5 mutants, resulting in a loss of NDH activity in these mutants. These results indicated that both NDF5 and NDF2 have essential functions in the stabilization of the NDH complex. We propose that NDF5 and NDF2 were acquired by land plants during evolution, and that in higher plants both NDF5 and NDF2 are critical to regulate NDH activity and each other's protein stability, as well as the stability of additional NDH subunits. PMID- 19122189 TI - Salivary acetylcholinesterase as a biomarker for organophosphate exposure. AB - BACKGROUND: Workers exposed to organophosphate (OP) pesticides are required to undergo periodic statutory medical surveillance in several countries. AIM: To study the relationship between serum, erythrocyte and saliva acetylcholinesterase (AChE) levels and to explore the use of salivary AChE as potential biomarker for OP exposure. METHODS: A cross-sectional study was conducted on 19 healthy adult male lead-exposed workers who were undergoing six monthly statutory medical examination. Passive drool saliva samples were collected from each worker. Each blood sample was tested for serum and erythrocyte AChE, and each saliva sample was tested for AChE. RESULTS: Among the 19 subjects, the mean (+/-standard deviation) of salivary, erythrocyte and serum AChE/cholinesterase were 22.7 (+/ 17.4), 17171 (+/-1467), 8861 (+/-1876) U/l, respectively. There was a moderate correlation between salivary and erythrocyte AChE (r = 0.42, P = 0.071), but not salivary and serum AChE (r = -0.17, P = 0.48). The level of AChE in saliva was approximately 1820 times lower than AChE in erythrocytes. CONCLUSION: It is probably not feasible to use saliva as a replacement for blood for the measurement of AChE levels. This is because of the much lower levels of AChE in saliva relative to erythrocytes, the weak correlation between the two measurements and the previously reported high intra-individual variation of salivary AChE. PMID- 19122190 TI - Audit of quality of diagnostic procedures for occupational asthma. AB - BACKGROUND: Previous studies have reported deficiencies in the quality of the diagnosis of occupational asthma. A low quality of diagnostic procedures means that the occupational cause of asthma is less likely to be revealed. AIMS: To assess the current quality of the diagnosis of occupational asthma before referral to a specialist occupational medicine centre. METHODS: The quality of diagnostic procedures was assessed by reviewing the files of 150 patients who were referred to the Finnish Institute of Occupational Health in 2003 with a suspicion of an occupational cause of their asthma. The quality indicators used were assessment of workplace exposures, spirometric studies, bronchodilator response, serial workplace measurements of peak expiratory flow (PEF) and the time since first symptoms to the final diagnosis. For each indicator, criteria to differentiate between sufficient and insufficient care were developed. RESULTS: Exposure assessments, spirometric studies and bronchodilator responses were performed in 92, 87 and 79% of cases in the total study group, respectively. Workplace measurements of PEF had been performed in 51% of the cases, and the quality of measurements was sufficient in 52%. Workplace exposures had been assessed significantly more often in occupational health care than in other health care units. The median time from the beginning of symptoms to the final diagnosis was 3.2 years. CONCLUSIONS: Although the diagnostic procedures were mostly of sufficient quality, the performance of serial measurements of PEF at the workplace and the time to diagnosis should be substantially improved. PMID- 19122191 TI - Accidental blood and body fluid exposure among doctors. AB - AIM: To study the epidemiology and time trends of blood and body fluids (BBF) exposures among hospital doctors. METHODS: A 3-year study was carried out using data from the Exposure Prevention Information Network of four teaching hospitals in the UK. RESULTS: One hundred and seventy-five cases of BBF exposures in doctors were reported over the 3-year study period. Eighty-one (46%) occurred in senior doctors and 94 (54%) in junior doctors. Junior doctors had a higher rate of BBF exposures compared to senior doctors: 13 versus 4 incidents per 100 person years, respectively (relative risk 3, 95% confidence interval 2-4). The most frequent setting for BBF exposures among senior doctors was the operating theatre/recovery (59%). Among junior doctors, it was the patient room (48%). The commonest original reason for use of sharps by junior doctors was the taking of blood samples (42%). Among senior doctors, it was suturing (41%). CONCLUSION: While ongoing training efforts need to be directed towards both junior and senior doctors, our data suggest that junior doctors are at higher risk of BBF exposures and may need particular attention in prevention strategies. An improvement in the safety culture in teaching hospitals can be expected to reduce the number of BBF exposures. PMID- 19122192 TI - Respiratory health of welders in a container yard, Sri Lanka. AB - BACKGROUND: The fumes and gases released during welding can lead to respiratory ill-health. AIMS: To assess prevalence of respiratory symptoms (RS) and respiratory function (RF) of welders in comparison to a control group (CG). METHODS: A cross-sectional study was conducted among welders and controls selected from office support staff of a medical faculty. RS were determined by administering a questionnaire and RF with the use of an electronic spirometer. RESULTS: Forty-one welders and 41 controls participated. Chronic bronchitis was significantly higher among welders (27%; n = 11) than in controls (7%; n = 3) with an odds ratio of 4.6 [95% confidence interval (CI): 1.1-23.3]. Forced vital capacity (FVC), forced expiratory volume in the first second of forced vital capacity (FEV(1.0)), forced mid-expiratory flow rate (FEF(25-75%)) and peak expiratory flow rate (PEFR) were slightly higher among welders (2.97 l, 2.6 l, 3.4 l/s and 339 l/min, respectively) than in the CG (2.79 l, 2.4 l, 3.38 l/s and 323 l/min, respectively), the differences of which were not statistically significant. On comparison of the observed values of welders with the predicted normal values, the observed FVC (2.97 versus 3.35 l, respectively) and PEFR (339 versus 538 l/min) had significantly lower values. CONCLUSIONS: Welders are at a higher risk of developing chronic bronchitis with non-impairment of lung function in comparison to the CG. PMID- 19122193 TI - Characterization and in vitro reaction properties of 19 unique hairpin telomeres from the linear plasmids of the lyme disease spirochete. AB - The genome of the Lyme disease pathogen Borrelia burgdorferi contains about a dozen linear DNA molecules that carry covalently closed hairpin telomeres as a specialized mechanism for dealing with the end-replication problem. The hairpin telomeres are generated from replicative intermediates through a two-step transesterification promoted by the telomere resolvase ResT. Although the genome of B. burgdorferi has been sequenced, the sequence of most telomeres has remained unknown because of difficulties in recovering and completely sequencing the covalently closed hairpin ends. In this study we report a new approach for the direct sequencing Borrelia telomeres and report the sequence, characterization, and in vitro reaction properties of 19 unique telomeres. Surprisingly, a variation of greater than 160-fold in the initial reaction rates of in vitro ResT mediated telomere resolution was observed between the most active and least active telomeres. Moreover, three of the hairpin telomeres were completely inactive in vitro, but their in vivo functionality was demonstrated. Our results provide important new information on the structure and function of the B. burgdorferi telomeres and suggest the possibility that factors besides the telomere resolvase ResT may influence the reaction in vivo and rescue those telomeres that are not functional in vitro with ResT alone. PMID- 19122194 TI - Novel mitochondrial complex II isolated from Trypanosoma cruzi is composed of 12 peptides including a heterodimeric Ip subunit. AB - Mitochondrial respiratory enzymes play a central role in energy production in aerobic organisms. They differentiated from the alpha-proteobacteria-derived ancestors by adding noncatalytic subunits. An exception is Complex II (succinate: ubiquinone reductase), which is composed of four alpha-proteobacteria-derived catalytic subunits (SDH1-SDH4). Complex II often plays a pivotal role in adaptation of parasites in host organisms and would be a potential target for new drugs. We purified Complex II from the parasitic protist Trypanosoma cruzi and obtained the unexpected result that it consists of six hydrophilic (SDH1, SDH2N, SDH2C, and SDH5-SDH7) and six hydrophobic (SDH3, SDH4, and SDH8-SDH11) nucleus encoded subunits. Orthologous genes for each subunit were identified in Trypanosoma brucei and Leishmania major. Notably, the iron-sulfur subunit was heterodimeric; SDH2N and SDH2C contain the plant-type ferredoxin domain in the N terminal half and the bacterial ferredoxin domain in the C-terminal half, respectively. Catalytic subunits (SDH1, SDH2N plus SDH2C, SDH3, and SDH4) contain all key residues for binding of dicarboxylates and quinones, but the enzyme showed the lower affinity for both substrates and inhibitors than mammalian enzymes. In addition, the enzyme binds protoheme IX, but SDH3 lacks a ligand histidine. These unusual features are unique in the Trypanosomatida and make their Complex II a target for new chemotherapeutic agents. PMID- 19122195 TI - Contribution of non-catalytic core residues to activity and regulation in protein kinase A. AB - Protein kinase A holoenzyme is comprised of two catalytic (C) and two regulatory (R) subunits which keep the enzyme in an inhibited state before activation by cyclic-AMP. The C-subunit folds into a conserved bi-lobal core flanked by N- and C-terminal tails. We report here characterization of a C-tail loss-of-function mutant, CF327A, and a related suppressor mutant, CF327A/K285P. Phe-327 is the only residue outside the kinase core that binds to the adenine ring of ATP, whereas Lys-285 is approximately 45 A away and lies in an AGC kinase-specific insert. The two mutations were previously identified from a yeast genetic screen, where the F327A mutation was unable to complement cell growth but mutation of K285P in the same allele rescued cell viability. We show that CF327A exhibits significant reduction in catalytic efficiency, which likely explains the observed loss-of-function phenotype. Interestingly, the additional K285P mutation does not restore kinase activity but reduces the inhibitory interaction of the double mutant with RII subunits. The additional K285P mutation, thus, helps to keep a low but uninhibited PKA activity that is sufficient for cell viability. The crystal structure of CF327A/K285P further reveals that recruitment of Phe-327 to the ATP binding pocket not only contributes to the hydrophobic pocket, as previously thought, but also recruits its flanking C-tail region to the kinase core, thereby concertedly positioning the glycine-rich loop and ATP for phosphoryl transfer. The study exemplifies two different ways for regulating cAMP dependent protein kinase activity through non-conserved residues and sheds light on the structural and functional diversity of the kinase family. PMID- 19122196 TI - Cyclical chromatin looping and transcription factor association on the regulatory regions of the p21 (CDKN1A) gene in response to 1alpha,25-dihydroxyvitamin D3. AB - The nuclear receptor vitamin D receptor (VDR) is known to associate with three vitamin D response element (VDREs)-containing regions within the CDKN1A (p21) gene region. Here we show in MDA-MB453 breast cancer cells that the natural VDR ligand 1alpha,25-dihydroxyvitamin D(3) causes cyclical transcription factor binding and chromatin looping of distal VDREs to the transcription start site (TSS) of the p21 gene, leading to cyclical accumulation of the p21 mRNA. At the chromatin level, association of the mediator protein MED1 precedes both the peaks of VDR binding to VDREs and phosphorylated RNA polymerase (p-Pol II) to the TSS. The loss of co-repressor NCoR1-histone deacetylase (HDAC) 3 complex and inhibitory chromatin looping from VDREs to the TSS are also initial events followed by increased acetylation of histone 3 at lysine 9 at the TSS prior to initiation of transcription. Simultaneous to VDR and p-Pol II peaks, chromatin loops between VDREs and the TSS are formed, and the lysine demethylase LSD1 and the histone acetyltransferase CBP are enriched in both regions. This is followed by a moderate peak in p21 transcript accumulation, repeated in cycles of 45-60 min. The transcript accumulation pattern is disturbed by siRNA inhibition of the mediator protein MED1, LSD1, NCoR1, or various HDACs, whereas CBP appears unnecessary for the response. Inhibition of MED1, HDAC4, or LSD1 by siRNA also attenuates ligand-induced chromatin looping. In conclusion, 1alpha,25 dihydroxyvitamin D(3) regulates p21 transcription by inducing cyclical chromatin looping that depends on both histone deacetylation and demethylation. PMID- 19122198 TI - Bacterial endotoxin stimulates adipose lipolysis via toll-like receptor 4 and extracellular signal-regulated kinase pathway. AB - Bacterial endotoxin/lipopolysaccharide elicits inflammatory responses and also elevates circulating levels of free fatty acids (FFAs) and impairs insulin sensitivity. Serum FFA elevation in acute endotoxemia has long been thought to be due to endotoxin dysregulating lipid disposal and counterregulatory hormones and cytokines. Here, we investigated the direct lipolysis effect of endotoxin in rodents and in isolated primary adipocytes. Endotoxin increases lipolysis in vivo in adipose tissues, elevates circulating FFA level, induces insulin resistance in rats, and directly stimulates chronic lipolysis in vitro in adipocytes. The lipolytic action of endotoxin is mediated via its lipid A moiety and is blocked by anti-endotoxin peptides. Neither adipocytokine secretion nor nuclear factor kappaB activation is involved in endotoxin-induced lipolysis. Different from catecholamine, endotoxin stimulates lipolysis without elevating cAMP production and activating protein kinase A and protein kinase C. Instead, endotoxin induces phosphorylation of Raf-1, MEK1/2, and ERK1/2. Upon inhibition of ERK1/2 but not JNK and p38 MAPK, endotoxin-stimulated lipolysis ceases. Endotoxin causes perilipin down-regulation and phosphorylation and increases the activity and protein levels of hormone-sensitive lipase and adipose triglyceride lipase but does not induce hormone-sensitive lipase translocation to intracellular lipid droplets. In TLR4 (Toll-like receptor 4)-deficient mice and adipocytes, endotoxin fails to increase in vivo and in vitro lipolysis. These findings suggest that endotoxin stimulates lipolysis via TLR4 and ERK1/2 signaling in adipocytes. The lipolytic action of endotoxin liberates FFA efflux from adipocytes to the bloodstream, which is a possible basis for systemic FFA elevation and insulin resistance in endotoxemia or Gram-negative bacterial infection. PMID- 19122197 TI - Calprotectin S100A9 calcium-binding loops I and II are essential for keratinocyte resistance to bacterial invasion. AB - Epithelial cells expressing calprotectin, a heterodimer of S100A8 and S100A9 proteins, are more resistant to bacterial invasion. To determine structural motifs that affect resistance to bacterial invasion, mutations were constructed in S100A9 targeting the calcium-binding loops I and II (E36Q, E78Q, E36Q,E78Q) and the C terminus (S100A9(1-99) and S100A9(1-112)), which contains putative antimicrobial zinc-binding and phosphorylation sites. The S100A8 and mutated S100A9 encoding plasmids were transfected into calprotectin-negative KB carcinoma cells. All transfected cells (except KB-sham) expressed 27E10-reactive heterodimers. In bacterial invasion assays with Listeria monocytogenes and Salmonella enterica serovar Typhimurium (Salmonella typhimurium), cell lines expressing S100A8 in complex with S100A9E36Q, S100A9E78Q, S100A9(1-99), or S100A9(1-112) mutants or the S100A9(1-114) (full-length) calprotectin resisted bacterial invasion better than KB-sham. When compared with KB-S100A8/A9(1-114), cells expressing truncated S100A9(1-99) or S100A9(1-112) with S100A8 also showed increased resistance to bacterial invasion. In contrast, glutamic acid residues 36 and 78 in calcium-binding loops I and II promote resistance in epithelial cells, because cells expressing S100A9E36Q,E78Q with S100A8 were unable to resist bacterial invasion. Mutations in S100A9 E36Q, E78Q were predicted to cause loss of the calcium-induced positive face in calprotectin, reducing interactions with microtubules and appearing to be crucial for keratinocyte resistance to bacterial invasion. PMID- 19122200 TI - Signalling pathway involving GULP, MAPK and Rac1 for SR-BI-induced phagocytosis of apoptotic cells. AB - Class B scavenger receptor type I (SR-BI) is a phosphatidylserine (PS) recognizing receptor of testicular Sertoli cells responsible for the phagocytosis of spermatogenic cells undergoing apoptosis. Here, we determined signal mediators that compose a signalling pathway for SR-BI-induced phagocytosis. Results of a yeast two-hybrid analysis and a cell-free binding assay indicated that SR-BI binds to engulfment adapter protein (GULP) using the C-terminal intracellular domain. A co-immunoprecipitation analysis showed the existence of a complex of GULP and SR-BI in cells prior to the activation of SR-BI by PS. A reduction of GULP expression in phagocytes decreased the SR-BI-mediated phagocytosis of apoptotic cells. Administration to phagocytes of PS-containing liposomes increased the levels of the GTP-bound form of Rac1 and the phosphorylated forms of mitogen-activated protein kinases (MAPK) p38 and extracellular signal-related kinase 1 and 2. Finally, lowering the expression of GULP abrogated MAPK phosphorylation, and the presence of MAPK inhibitors reduced the level of GTP bound Rac1 in PS-activated phagocytes. These results collectively suggested the following signalling pathway for the SR-BI-induced phagocytosis: (i) PS recognizing SR-BI activates associated GULP; (ii) activated GULP induces MAPK phosphorylation; (iii) activated MAPK increases GTP-bound Rac1; and (iv) activated Rac1 induces a rearrangement of the actin cytoskeleton. PMID- 19122199 TI - Retinoic acid-induced gene-1 (RIG-I) associates with the actin cytoskeleton via caspase activation and recruitment domain-dependent interactions. AB - The actin cytoskeleton serves as a barrier that protects mammalian cells from environmental pathogens such as bacteria, fungi, and viruses. Several components of antimicrobial signaling pathways have been shown to associate directly with the actin cytoskeleton, indicating that the cytoskeleton may also serve as a platform for immune-associated molecules. Here we report that retinoic acid induced gene-I (RIG-I), an important viral RNA recognition molecule, is associated with the actin cytoskeleton and localizes predominantly to actin enriched membrane ruffles in non-polarized epithelial cells. Subcellular localization and fractionation experiments revealed that the association between RIG-I and the actin cytoskeleton was mediated by its N-terminal caspase activation and recruitment domains (CARDs), which were necessary and sufficient to induce cytoskeletal association. We also show that RIG-I plays a role in cellular migration, as ectopic expression of RIG-I enhanced cellular migration in a wound healing assay and depletion of endogenous RIG-I significantly reduced wound healing. We further show that in both cultured intestinal epithelial cells (IEC) and human colon and small intestine biopsies, RIG-I is enriched at apico lateral cell junctions and colocalizes with markers of the tight junction. Depolymerization of the actin cytoskeleton in polarized IEC led to the rapid relocalization of RIG-I and to the induction of type I interferon signaling. These data provide evidence that RIG-I is associated with the actin cytoskeleton in non-polarized epithelial cells and with the junctional complex in polarized IECs and human intestine and colon biopsies and may point to a physiological role for RIG-I in the regulation of cellular migration. PMID- 19122201 TI - The eighth fibronectin type III domain of protein tyrosine phosphatase receptor J influences the formation of protein complexes and cell localization. AB - Regulation of receptor-type phosphatases can involve the formation of higher order structures, but the exact role played in this process by protein domains is not well understood. In this study we show the formation of different higher order structures of the receptor-type phosphatase PTPRJ, detected in HEK293A cells transfected with different PTPRJ expression constructs. In the plasma membrane PTPRJ forms dimers detectable by treatment with the cross-linking reagent BS(3) (bis[sulfosuccinimidyl]suberate). However, other PTPRJ complexes, dependent on the formation of disulfide bonds, are detected by treatment with the oxidant agent H(2)O(2) or by a mutation Asp872Cys, located in the eighth fibronectin type III domain of PTPRJ. A deletion in the eighth fibronectin domain of PTPRJ impairs its dimerization in the plasma membrane and increases the formation of PTPRJ complexes dependent on disulfide bonds that remain trapped in the cytoplasm. The deletion mutant maintains the catalytic activity but is unable to carry out inhibition of proliferation on HeLa cells, achieved by the wild type form, since it does not reach the plasma membrane. Therefore, the intact structure of the eighth fibronectin domain of PTPRJ is critical for its localization in plasma membrane and biological function. PMID- 19122202 TI - Closed complex of the D-3-hydroxybutyrate dehydrogenase induced by an enantiomeric competitive inhibitor. AB - D-3-Hydroxybutyrate dehydrogenase (HBDH) from Pseudomonas fragi showed a strict stereospecificity to the d-enantiomer of 3-hydroxybutyrate (d-3-HB) as a substrate. The l-enantiomer acts as a competitive inhibitor, with a K(i) value comparable to the K(m) value for d-3-HB. We have determined the crystal structures of the ternary complex of HBDH-NAD(+)-l-3-HB and the binary complex of HBDH-NAD(+). The former structure showed a so-called closed-form conformation, which is considered an active form for catalysis, while the latter stayed mostly in a open-form conformation. The determined structures along with the site directed mutagenesis confirmed the substrate recognition mechanism that we proposed previously. The hydrogen bonding interaction between Gln196, located in the moving helix, and the carboxyl group of the substrate/inhibitor is important for the stable ternary complex formation. Finally, the crystal structures of the Thr190 mutants, T190S and T190A, indicate that the Thr190 is a key residue for the open-closed conformational change. T190S retained 37% of the activity. In T190A, however, the activity decreased to 0.1% that of the wild-type enzyme. Fixing the position of the hydroxyl group of Thr190 to form hydrogen bonds to the pyrophosphate moiety and the carboxamide of NAD(+) seems to be a significant factor for the open-closed conformational change. PMID- 19122203 TI - Functional expression of miraculin, a taste-modifying protein in Escherichia coli. AB - Miraculin isolated from red berries of Richadella dulcifica, a native shrub of West Africa, has the unusual property of modifying a sour taste into a sweet one. This homodimer protein consists of two glycosylated polypeptides that are cross linked by a disulfide bond. Recently, functional expression of miraculin was reported in host cells with the ability to glycosylate proteins, such as lettuce, tomato and the microbe Aspergillus oryzae, but not Escherichia coli. Thus, a question remains as to whether glycosylation of miraculin is essential for its taste-modifying properties. Here we show that recombinant miraculin expressed in E. coli has taste-modifying properties as a homodimer, not as a monomer, indicating that glycosylation is not essential for the taste-modifying property. PMID- 19122204 TI - Concerted effects of two activator modules on the group I ribozyme reaction. AB - Group I intron ribozymes have a modular architecture and structural elements essential for catalysis. The elements are located in the conserved modular domain P3-P7 that is stabilized by another conserved module, P4-P6. It has been reported that artificial modules can complement the function of the native P4-P6. To exploit the modular architecture of group I ribozyme, we have constructed a hybrid ribozyme by attaching an artificial activator module to the wild-type T4 td ribozyme. Kinetic analysis of the hybrid ribozyme revealed that the artificial module and P4-P6 have unusual positive and negative concerted effects in activating the ribozyme. PMID- 19122205 TI - Identification and functional characterization of rat riboflavin transporter 2. AB - We have newly identified rat riboflavin transporter 2 (rRFT2) and its human orthologue (hRFT2), and carried out detailed functional characterization of rRFT2. The mRNA of rRFT2 was highly expressed in jejunum and ileum. When transiently expressed in human embryonic kidney (HEK) 293 cells, rRFT2 could transport riboflavin efficiently. Riboflavin transport mediated by rRFT2 was Na(+)-independent but moderately pH-sensitive, being more efficient in acidic conditions than in neutral and basic conditions. Kinetic analysis indicated that rRFT2-mediated riboflavin transport was saturable with a Michaelis constant (K(m)) of 0.21 microM. Furthermore, it was specifically and strongly inhibited by lumiflavin, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), and to a lesser extent by amiloride. Such ability to transport riboflavin in a specific manner, together with its high expression in the small intestine, indicates that RFT2 may play a role in the intestinal absorption of riboflavin. PMID- 19122206 TI - In vitro and in vivo interactions of ferredoxin-NADP+ reductases in Pseudomonas putida. AB - Ferredoxin-NADP(+) reductase (Fpr) is known to control NADP(+)/NADPH pool in proteobacteria. There is only one fpr gene present in most proteobacteria, but Pseudomonas putida has two Fprs (FprA and FprB). We elucidated the functional relationships between the two types of Fpr and their electron transport partners [ferredoxin (Fd) and flavodoxin (Fld)] by cloning, expressing and preparing these proteins in various combinations and assessing their properties in vitro and in vivo using biochemical assays, the Far-western analysis, the yeast two-hybrid assay and structural molecular modelling. Both of the Fprs have a lower K(m) value for NADPH than for NADH in the diaphorase assays. With NADH as electron donor, FprB also has a high specific constant (k(cat)/K(m)) in the diaphorase assay. The catalytic efficiency of FprA is higher when Fld is present as its redox partner, compared to the kinetics observed with other electron transport partners in a NADPH-dependent cytochrome c reduction assay. The highest specific constant (k(cat)/K(m)) of FprB was observed in the presence of FdA. FprB's K(m) value and catalytic activity (k(cat)) with NADH were significant in cytochrome c reduction assays. Strong kinetic interactions of Fprs with their redox partners were also demonstrated by homology modelling, the Far-western analysis and the in vivo yeast two-hybrid system. This study demonstrates for the first time that Fprs in P. putida function as diaphorase, Fd/Fld reductases and determines their preferred redox partner in vivo and in vitro. PMID- 19122207 TI - Turnip mosaic virus genome-linked protein VPg binds C-terminal region of cap bound initiation factor 4E orthologue without exhibiting host cellular specificity. AB - To investigate the binding specificity of turnip mosaic virus (TuMV) viral protein-genome linked (VPg) with translation initiation factor 4E, we evaluated here the kinetic parameters for the interactions of human eIF4E, Caenorhabditis elegans IFE-3 and IFE-5 and Arabidopsis eIFiso4E, by surface plasmon resonance (SPR). The results indicated that TuMV VPg does not show a binding preference for Arabidopsis eIFiso4E, even though it is from a host species whereas the other eIF4E orthologues are not. Surprisingly, the effect of m(7)GTP on both the rate constants and equilibrium binding constants for the interactions of VPg differed for the four eIF4E orthologues. In the case of eIFiso4E and IFE-3, m(7)GTP increased k(on), but for eIF4E and IFE-5, it decreased k(on). To provide insight into the structural basis for these differences in VPg binding, tertiary structures of the eIF4E orthologues were predicted on the basis of the previously determined crystal structure of m(7)GpppA-bound human eIF4E. The results suggested that in cap-bound eIF4E orthologues, the VPg binds to the C-terminal region, which constitutes one side of the entrance to the cap-binding pocket, whereas in the cap-free state, VPg binds to the widely opened cap-binding pocket and its surrounding region. The binding of VPg to the C-terminal region was confirmed by the SPR analyses of N- or C-terminal residues-deleted eIF4E orthologues. PMID- 19122208 TI - Repression of estrogen receptor alpha by CDK11p58 through promoting its ubiquitin proteasome degradation. AB - Estrogen receptor alpha (ERalpha) is a ligand-dependent transcription factor that mediates physiological responses to 17beta-estradiol (E(2)). These responses of cells to estrogen are regulated in part by degradation of ERalpha. In this report, we found that CDK11(p58) repressed ERalpha transcriptional activity. And we further demonstrated that ERalpha protein level was down-regulated by CDK11(p58) in mammalian cells in a ligand independent manner. This effect could be abrogated by treatment with proteasome inhibitor MG132. Our results indicated that the ubiquitin/proteasome-mediated degradation of ERalpha was promoted by CDK11(p58). Furthermore, the interaction between ERalpha and CDK11(p58) was detected. This interaction was necessary for the polyubiquitination and degradation of ERalpha. On the contrary, the other isoform of CDK11, CDK11(p110) and the kinase dead mutant of CDK11(p58), D224N, did not associate with ERalpha and failed to reduce the ERalpha protein level. These data identified a new negative regulatory protein of ERalpha and provided a new pathway by which CDK11(p58) negatively regulated cells. PMID- 19122209 TI - TTP at Ser245 phosphorylation by AKT is required for binding to 14-3-3. AB - Transferrin receptor trafficking protein (TTP) is a key molecule for selective internalization of the transferrin receptor (Tf-R) through endocytic protein complexes. To identify the proteins that directly regulate TTP, we performed a yeast two-hybrid analysis and identified 14-3-3, which can modulate the activation state of target proteins. Subsequent analyses demonstrated that TTP directly binds to multiple 14-3-3 isotypes via its Ser(245) residue (Ser(246) in human) and that these proteins are associated at the plasma membrane. Ser(245) was also found to be a substrate for AKT and the resulting Ser(245) phosphorylation induced the TTP-14-3-3 interaction. Exposure to hydrogen peroxide rapidly enhanced this association in an ovarian cell line. These results suggest that TTP Ser(245) is the principal target for the modulation of this protein via the AKT signalling cascade. PMID- 19122210 TI - Why depression is associated with increased risk towards premature labor. PMID- 19122212 TI - DXRaySMCS: a user-friendly interface developed for prediction of diagnostic radiology X-ray spectra produced by Monte Carlo (MCNP-4C) simulation. AB - In this work, the general purpose Monte Carlo N-particle radiation transport computer code (MCNP-4C) was used for the simulation of X-ray spectra in diagnostic radiology. The electron's path in the target was followed until its energy was reduced to 10 keV. A user-friendly interface named 'diagnostic X-ray spectra by Monte Carlo simulation (DXRaySMCS)' was developed to facilitate the application of MCNP-4C code for diagnostic radiology spectrum prediction. The program provides a user-friendly interface for: (i) modifying the MCNP input file, (ii) launching the MCNP program to simulate electron and photon transport and (iii) processing the MCNP output file to yield a summary of the results (relative photon number per energy bin). In this article, the development and characteristics of DXRaySMCS are outlined. As part of the validation process, output spectra for 46 diagnostic radiology system settings produced by DXRaySMCS were compared with the corresponding IPEM78. Generally, there is a good agreement between the two sets of spectra. No statistically significant differences have been observed between IPEM78 reported spectra and the simulated spectra generated in this study. PMID- 19122211 TI - Incidence and risk factors of bladder injuries during laparoscopic hysterectomy indicated for benign uterine pathologies: a 14.5 years experience in a continuous series of 1501 procedures. AB - BACKGROUND: Laparoscopic hysterectomy is indicated as an alternative to laparotomy when the vaginal route is potentially difficult because of an immobile uterus and a poor vaginal accessibility. The aim of this study was to evaluate the rate, the risk factors for bladder injuries in a series of 1501 laparoscopic hysterectomies indicated for benign uterine pathologies. METHODS: This study was conducted retrospectively from January 1993 to 2000 and prospectively from 2001 to July 2007.The indications, patients' characteristics and complications were recorded. The overall rate of bladder injuries, the comparison of means (t test) and percentages (exact chi(2) test) between the cases and the population with no injury, the odd ratios (OR) and multivariate analysis were performed using the statistical package for the social sciences software. RESULTS: The rate of bladder injuries was 1% (15 patients). Risks factors were previous Caesarian section [OR: 4.33, 95% confidence interval (CI): 1.53-12.30] and previous laparotomy (OR: 4.69, 95% CI: 1.59-13.8). The rate of injury decreases with the surgeons' experience and reaches a plateau of 0.4% after 100 hysterectomies performed. CONCLUSIONS: The rate of bladder injury during total laparoscopic hysterectomy is low and decreases with the surgeon's experience. Bladder injury is not linked to an increase of post-operative morbidity when recognized and repaired during the same laparoscopic procedure. The comparison with other routes of hysterectomies should take into account these risk factors. PMID- 19122213 TI - O-glycan inhibitors generate aryl-glycans, induce apoptosis and lead to growth inhibition in colorectal cancer cell lines. AB - Our studies provide direct evidence that O-glycosylation pathways play a role in the regulation of cell growth through apoptosis and proliferation pathways. A series of small molecular weight analogs of the GalNAc-alpha-1-O-serine/threonine structure based on 1-benzyl-2-acetamido-2-deoxy-alpha-O-d-galactopyranoside have been synthesized and tested in the human colorectal cancer cell lines PC/AA/C1/SB10C and HCA7/C29. Three inhibitors, 1-benzyl-2-acetamido-2-deoxy-alpha O-D-galactopyranoside, and the corresponding 2-azido- and C-glycoside analogs were screened in these colorectal cancer cell lines at 0.5 mM and showed induction of apoptosis and downregulation of proliferation. Treatment of both cell lines with inhibitors led to changes in glycosylation detected with peanut lectin. The inhibition of glycosyltransferase activity in cell homogenates from human colorectal mucosal cells and cultured cell lines could be shown. The competitive action of the inhibitors resulted in the intracellular formation of 28 aryl-glycan products which were identified by MALDI and electrospray mass spectroscopy. The structures showed a differential pattern for each of the inhibitors in both cell lines. Gene array analysis of the glycogenes illustrated a pattern of glycosyltransferases that matched the glycan structures found in glycoproteins and aryl-glycans formed in the PC/AA/C1/SB10C cells; however, there was no action of the three inhibitors on glycogene transcript levels. The inhibitors act at both intermediary metabolic and genomic levels, resulting in altered protein glycosylation and aryl-glycan formation. These events may play a part in growth arrest. PMID- 19122214 TI - Effect of left-sided valvular regurgitation on mortality and causes of death among a population of middle-aged and older horses. AB - The effect of left-sided valvular regurgitation (LSVR) on the mortality of middle aged and older horses was investigated in a prospective cohort study involving 19 yards and 1153 horses. The horses were examined to determine whether they had a cardiac murmur and its type, and their age, sex, breed type and occupation were recorded. They were followed up at intervals of two years by postal questionnaire, and after four years information on 773 horses was available. There was no significant difference in the mortality of the horses with and without LSVR, but small horses had a significantly higher risk of having LSVR than small ponies (odds ratio [OR] 2.33), and older horses were slightly more likely to have LSVR than young horses (OR 1.07). Twenty-nine per cent of the deaths reported by the owners were due to orthopaedic problems, 23.3 per cent to gastrointestinal problems, and only 7.9 per cent to cardiovascular problems. Orthopaedic problems were the main cause of death in the horses, and gastrointestinal problems were the main cause of death in the ponies. PMID- 19122215 TI - Ultrasonographic evaluation of reticular motility in 144 cattle with vagal indigestion. AB - The reticular motility in 144 cattle with vagal indigestion was evaluated by ultrasonography; 97 had proximal functional stenosis (reticulo-omasal stenosis) and 47 had distal functional stenosis (pyloric stenosis). A 3.5 MHz linear transducer was used to measure the frequency, amplitude, duration and speed of the reticular contractions per three minutes while the animals were standing and unsedated. The mean (sd) number of contractions per three minutes was 4.3 (2.09) with a range from 0 to 12; nine of the cattle had one or two contractions, 17 had three contractions, 59 had four or five contractions, 37 had more than five contractions and 12 had reticular atony. The cattle with proximal functional stenosis had 4.6 (2.01) contractions per three minutes, significantly more (P<0.05) than those with distal functional stenosis (3.6 [2.16]). The position, contour and size of the reticulum, the amplitude and speed of the contractions and the area surrounding the reticulum did not differ significantly between the two groups. PMID- 19122216 TI - Comparison of CO(2) laser and 4.0 MHz radiosurgery for making incisions in the skin and muscles of green iguanas (Iguana iguana). AB - Four green iguanas scheduled for euthanasia were used to compare the extent of collateral tissue damage associated with CO(2) laser and 4.0 MHz radiosurgery. The iguanas were anaesthetised and a series of three skin and three muscle incisions was made by 4.0 MHz radiosurgery (0.18 mm wire electrode, 25 W, cut mode) and CO(2) laser (0.3 mm ceramic tip, 15 W focused beam super-pulse mode), and three incisions were made with a scalpel blade as controls. Following euthanasia, a total of 60 skin and 36 muscle sections were evaluated histologically. Radiosurgery and the laser both produced bloodless incisions, but radiosurgery caused significantly less collateral tissue damage in the skin (307 [97] v 386 [108] microm) and the muscle (18 [7] v 91 [15] microm). PMID- 19122217 TI - Prevalence of Chlamydophila psittaci-positive cloacal PCR tests in wild avian casualties in the UK. PMID- 19122218 TI - Fatal mesenteric lymphadenitis in cattle caused by Listeria monocytogenes. PMID- 19122219 TI - Unilateral uterine torsion associated with haematometra and cystic endometrial hyperplasia in a bitch. PMID- 19122220 TI - Uterus bicorpora bicollis as a possible cause of infertility in a mare. PMID- 19122221 TI - Atypical clinical presentation of coinfection with Ehrlichia, Babesia and Hepatozoon species in a dog. PMID- 19122222 TI - Cryptosporidium hominis in a goat and a sheep in the UK. PMID- 19122223 TI - Disease threats from travelling pets. PMID- 19122224 TI - Expiry dates on opened injectables. PMID- 19122225 TI - End of the Older Cattle Disposal Scheme. PMID- 19122226 TI - Extramural studies for undergraduates. PMID- 19122228 TI - Veterinary nurse training. PMID- 19122229 TI - Breeding of pedigree dogs. PMID- 19122239 TI - ERdj5 sensitizes neuroblastoma cells to endoplasmic reticulum stress-induced apoptosis. AB - Down-regulation of the unfolded protein response (UPR) can be therapeutically valuable in cancer treatment, and endoplasmic reticulum (ER)-resident chaperone proteins may thus be targets for developing novel chemotherapeutic strategies. ERdj5 is a novel ER chaperone that regulates the ER-associated degradation of misfolded proteins through its associations with EDEM and the ER stress sensor BiP. To investigate whether ERdj5 can regulate ER stress signaling pathways, we exposed neuroblastoma cells overexpressing ERdj5 to ER stress inducers. ERdj5 promoted apoptosis in tunicamycin, thapsigargin, and bortezomib-treated cells. To provide further evidence that ERdj5 induces ER stress-regulated apoptosis, we targeted Bcl-2 to ER of ERdj5-overexpressing cells. Targeting the Bcl-2 to ER prevented the apoptosis induced by ER stress inducers but not by non-ER stress apoptotic stimuli, suggesting induction of ER stress-regulated apoptosis by ERdj5. ERdj5 enhanced apoptosis by abolishing the ER stress-induced phosphorylation of eukaryotic translation initiation factor 2alpha (eIF2alpha) and the subsequent translational repression. ERdj5 was found to inhibit the eIF2alpha phosphorylation under ER stress through inactivating the pancreatic endoplasmic reticulum kinase. The compromised integrated stress response observed in ERdj5-overexpressing ER-stressed cells due to repressed eIF2alpha phosphorylation correlated with impaired neuroblastoma cell resistance under ER stress. These results demonstrate that ERdj5 decreases neuroblastoma cell survival by down-regulating the UPR, raising the possibility that this protein could be a target for anti-tumor approaches. PMID- 19122238 TI - Transport of maternal cholesterol to the fetus is affected by maternal plasma cholesterol concentrations in the golden Syrian hamster. AB - The fetus has a high requirement for cholesterol and synthesizes cholesterol at elevated rates. Recent studies suggest that fetal cholesterol also can be obtained from exogenous sources. The purpose of the current study was to examine the transport of maternal cholesterol to the fetus and determine the mechanism responsible for any cholesterol-driven changes in transport. Studies were completed in pregnant hamsters with normal and elevated plasma cholesterol concentrations. Cholesterol feeding resulted in a 3.1-fold increase in the amount of LDL-cholesterol taken up by the fetus and a 2.4-fold increase in the amount of HDL-cholesterol taken up. LDL-cholesterol was transported to the fetus primarily by the placenta, and HDL-cholesterol was transported by the yolk sac and placenta. Several proteins associated with sterol transport and efflux, including those induced by activated liver X receptor, were expressed in hamster and human placentas: NPC1, NPC1L1, ABCA2, SCP-x, and ABCG1, but not ABCG8. NPC1L1 was the only protein increased in hypercholesterolemic placentas. Thus, increasing maternal lipoprotein-cholesterol concentrations can enhance transport of maternal cholesterol to the fetus, leading to 1) increased movement of cholesterol down a concentration gradient in the placenta, 2) increased lipoprotein secretion from the yolk sac (shown previously), and possibly 3) increased placental NPC1L1 expression. PMID- 19122240 TI - Pin1 down-regulates transforming growth factor-beta (TGF-beta) signaling by inducing degradation of Smad proteins. AB - Transforming growth factor-beta (TGF-beta) is crucial in numerous cellular processes, such as proliferation, differentiation, migration, and apoptosis. TGF beta signaling is transduced by intracellular Smad proteins that are regulated by the ubiquitin-proteasome system. Smad ubiquitin regulatory factor 2 (Smurf2) prevents TGF-beta and bone morphogenetic protein signaling by interacting with Smads and inducing their ubiquitin-mediated degradation. Here we identified Pin1, a peptidylprolyl cis-trans isomerase, as a novel protein binding Smads. Pin1 interacted with Smad2 and Smad3 but not Smad4; this interaction was enhanced by the phosphorylation of (S/T)P motifs in the Smad linker region. (S/T)P motif phosphorylation also enhanced the interaction of Smad2/3 with Smurf2. Pin1 reduced Smad2/3 protein levels in a manner dependent on its peptidyl-prolyl cis trans isomerase activity. Knockdown of Pin1 increased the protein levels of endogenous Smad2/3. In addition, Pin1 both enhanced the interaction of Smurf2 with Smads and enhanced Smad ubiquitination. Pin1 inhibited TGF-beta-induced transcription and gene expression, suggesting that Pin1 negatively regulates TGF beta signaling by down-regulating Smad2/3 protein levels via induction of Smurf2 mediated ubiquitin-proteasomal degradation. PMID- 19122241 TI - Reservoir drug delivery. PMID- 19122242 TI - Designing and performing large multicenter randomized clinical trials in India: what do we need? PMID- 19122243 TI - Clopidogrel resistance: chemical curiosity or clinical enigma? PMID- 19122244 TI - Infective endocarditis: prognostic stratification and indications for valve surgery. AB - Significant advances have been made in the management of infective endocarditis with the advent of antimicrobial therapy, surgery and transesophageal echocardiography, bringing hope to patients suffering from this once uniformly fatal illness. However, recent studies show that the mortality from infective endocarditis is still in the order of 20-30%. The prognostic determinants for adverse outcomes and the indications for surgical therapy in patients with infective endocarditis have recently received renewed attention. Our current understanding of these controversial issues will be reviewed in this paper. PMID- 19122245 TI - Current perspectives on coronary revascularization in the diabetic patient. AB - In the last few decades, the increase in the prevalence of type 2 diabetes mellitus has reached epidemic proportions in the Western world. A similar, though delayed, pattern is seen in developing countries. Chronic hyperglycemia, dyslipidemia, and insulin resistance have been associated with an accelerated form of atherogenesis, characterized by a prothrombotic state, enhanced inflammation, and endothelial dysfunction. Diabetic patients undergoing coronary revascularization have worse outcomes compared to nondiabetic individuals, both in the setting of percutaneous coronary intervention (PCI) and coronary artery bypass surgery (CABG). Though subgroup analyses of randomized trials and registries have suggested that CABG is superior to PCI in diabetic patients with multivessel disease, the results of ongoing large randomized trials focusing for the first time on diabetic patients should be awaited in order to establish the optimal revascularization strategy in this patient population. In the setting of acute coronary syndromes, diabetic patients are also at high short- and long-term risk for cardiovascular morbidity and mortality. Despite deriving greater benefit than nondiabetic counterparts from aggressive management, including early revascularization and potent platelet inhibition, diabetic patients are often undertreated. PMID- 19122246 TI - Impact of left ventricular function on early outcomes after arterial switch for D transposition of great arteries with intact ventricular septum. AB - OBJECTIVE: The aim of this study was to assess the utility of tissue Doppler echocardiography in evaluating the pre-operative left ventricular systolic function and in turn its impact on early post-operative outcomes following arterial switch operation for transposition of great arteries with intact ventricular septum. BACKGROUND: Pre-operative left ventricular function is an important determinant of outcomes following arterial switch. METHODS: We studied 19 patients with transposition and intact septum who had undergone one stage arterial switch operation. All had a pre-operative echocardiogram. RESULTS: The left ventricle was adequate in 15 infants as per the dimensions and shape for age. The early outcomes of surgery were assessed on the basis of their pre operative tissue Doppler. There were 7 (40%) patients with reduced S wave velocities (2.62 +/- 0.84 cm/sec) and lower isovolumetric myocardial acceleration (1.2 +/- 0.5 m/sec(2)) reflecting systolic dysfunction. Those patients with reduced S wave velocities and isovolumetric acceleration correlated with poor early surgical outcomes in terms of longer duration of ventilation ( p< 0.001), longer duration of inotropes ( p< 0.00001) and higher mortality ( p < 0.001). CONCLUSION: The tissue Doppler echocardiography is a more sensitive modality to evaluate ventricular performance than the 2D-echo. It can be used as a sensitive predictor of outcomes following arterial switch. PMID- 19122247 TI - Cardiac dimensions and function in children with obesity. AB - BACKGROUND: Obesity can cause alterations in cardiac dimensions and function. Cardiac dysfunction during childhood may affect the quality of life in adulthood. This study evaluated left ventricular (LV) dimensions, systolic function and left ventricular myocardial performance index (LMPI) in children with obesity. METHODS AND RESULTS: Thirty-three obese children with mean age of 9.8 +/- 2.4 years, weight 61.3 +/- 20.8 kg, BMI 29.5 +/- 5.8 kg/m2 and percentage of actual weight to ideal body weight for height (%IBW) 170 +/- 25%, underwent echocardiography to assess LV dimensions, systolic and global functions. There were 2, 14 and 17 children with mild (160%IBW), respectively. The mean ratio of left ventricular end-diastolic dimension (LVEDD) to predicted LVEDD expressed in percentage (%LVEDD) was 98.3 +/ 7.8%, the left ventricular shortening fraction (LVFS) was 37.5 +/- 4.9% and the left ventricular ejection fraction (LVEF) was 67.5 +/- 5.9%. All were within normal range except that 2 children (6%) had mild LV dilatation. The mean LMPI was 0.35 +/- 0.08. However, 11 children (33%) had abnormal LMPI (>0.4). The severity of obese children did not correlate with the global LV dysfunction. CONCLUSION: The left ventricular dimensions and systolic function in children with obesity were essentially normal. LMPI which indicates LV global function was found to be abnormal in 33% of children with obesity and may be used to do early detection of LV global dysfunction. PMID- 19122248 TI - Risk factors of coronary heart disease. AB - Extensive clinical and statistical studies have identified several factors that increase the risk of coronary heart disease(CHD). This study is another attempt to identify the factors that increase the risk for CHD as it is an extremely important area in health sciences. Utilizing the Framingham data about a wide variety of risk factors associated with CHD,researchers are now working to develop better tools to assess overall risk. The incidence of CHD was recorded along with patients age, sex and cholesterol level in the study. Logistic regression was used to model the log odds of developing CHD as a function of cholesterol category (0: < 190 mg/100 ml, 1: 190-219 mg/ml, 2: 220-249 mg/100ml, 37 colon; 250 mg/100ml), adjusting for age, sex and their interaction. The analysis showed that serum cholesterol level is a risk factor of coronary heart disease, but its effect is modified by the age category of the subjects. Sex is also associated with CHD, and moreover the effect of age on the 12-year incidence of CHD is gender-dependent. PMID- 19122249 TI - Profile and prevalence of clopidogrel resistance in patients of acute coronary syndrome. AB - Recently reports of a variable platelet response to aspirin and potential resistance to therapy have emerged with thienopyridines group of drugs. However the data available on clopidogrel resistance is scarce. The present study was initiated to prospectively evaluate the prevalence of clopidogrel resistance in patients of acute coronary syndrome(ACS) presently on dual anti platelet therapy by using an established method of optical platelet aggregation. We studied 39 patients of ACS, who were on clopidogrel 300 mg bolus followed by 75 mg per day for 3 days along with aspirin 325 mg per day. Fasting blood samples were assessed using optical platelet aggregation (Chronolog Corp, USA). Clopidogrel resistance was defined as <10% decrease from baseline in platelet aggregation. Clopidogrel semi-responders were defined as 10-29% ( <30%) decrease from baseline in platelet aggregation. Clopidogrel non-responders were defined as a composite of resistant and semi-responders. A baseline mean platelet aggregation obtained from 18 healthy subjects was 63.8 +/- 14.75% with 5 mu and 68.8 +/-13.91% with 10 mu of Adenosine Diphosphate. Hence, the definition of clopidogrel resistance was set as aggregation of >57% with 5 mu ADP and >61.9% with 10 mu ADP (< 10% decrease from baseline). The definition of clopidogrel semi-responder was set as aggregation of >or=45% with 5 mu ADP and >or=48% with 10 mu ADP (10-29% decrease from baseline). The mean platelet aggregation with 5 mu and 10 mu of Adenosine Diphosphate in the patient group was 30.77 +/- 17.19% and 35.71 +/- 17.0% respectively. Based on these criteria, 2.54% patients were found to be clopidogrel resistant, 12.7% were clopidogrel semi-responders and 84.7% were clopidogrel responders. On comparison of clopidogrel responders with non-responders, females ( p=0.07) and patients with higher serum triglyceride levels (p=0.08), had a trend to be more inclined towards clopidogrel non-responders. All other parameters tested namely age, smoking, diabetes, hypertension, obesity, cholesterol, hemoglobin, platelet count, ejection fraction and concurrent drug intake did not show any statistically significant difference among the groups. CONCLUSIONS: This study shows that clopidogrel resistant and clopidogrel semi-responders do occur in Indian patients with ACS and there are no reliable clinical predictors for this condition. The diagnosis therefore relies primarily on laboratory tests. PMID- 19122250 TI - Prevalence of coronary artery disease and coronary risk factors in an urban population of Tirupati. AB - AIMS AND OBJECTIVES: To determine the prevalence of coronary heart disease and its risk factors in the urban population of Tirupati. METHODS: The urban population of Tirupati is spread into 20 wards. Thirteen wards were selected randomly and systematic random sampling was done to identify the sampling unit household. A total of 1519 subjects (539 males and 980 females) 20 years of age formed the sample of the study. Demographic (age, sex, occupation, education and income), behavioral (smoking), anthropometric (height, weight, waist and hip circumferences) and physiological (blood pressure) parameters were noted and recorded and biochemical parameters (serum glucose, total cholesterol, triglycerides, HDL cholesterol levels) were assayed. Coronary artery disease was diagnosed based on history and ECG changes suggestive of ST-segment depression (Minnesota 4-1 to 4-2) or Q wave changes (Minnesota codes 1-1-1 to 1-1-7) or T wave changes (Minnesota codes 5-1 to 5-3). RESULTS: The overall prevalence in the study population was 12.63 (192 cases). In males it was 6.86 (37 cases) and in females 15.81 (155 cases). The major risk factors in this population were central obesity 1003 (66%) [male 320 (59.4%) and females 683 (41.12%)], low HDL cholesterol 709 (46.67%) [male 284 (52.69%) and females 425 (43.36%)], obesity 588 (38.07%) [males 187 (34.32%) and females 403 (41.12%)], high triglyceride levels (>or= 150 mg/dl) 444 (29.22%)[(males 173 (32.09%) and females 272 (27.75%)], high cholesterol levels (>or= 200mg/dl) [(males 127 (23.56%) and females 278 (28.36%)] and hypertension 396 (26.06) [(males 149 (27.64%) and females 247 (25.20%)]. Around a fraction of subjects had metabolic syndrome 371 (24.42%) [(males 90 (16.69%) and females 281 (28.64%)]. In multivariate regression analysis, age (OR 1.05 95% CI 1.01-1.09 p<0.01) and high serum LDL cholesterol level (OR 4.62 95% CI 2.29-9.32 p<0.001) in males and serum triglycerides (OR 1.003 95% CI 1.000-1.005) in females were associated with CAD. CONCLUSIONS: Association of age, high LDL cholesterol in males and triglycerides in females with CAD shows the importance of these risk factors in this population. This was only a cross-sectional study done at one point of time. Clinical importance of these risk factors needs to be tested in the longitudinal study. PMID- 19122251 TI - Cobalt chromium stent with antiproliferative for restenosis trial in India (COSTAR I). AB - BACKGROUND: The CoStar stent is a novel cobalt chromium stent designed specifically for drug delivery. The COSTAR I trial represents the first-in-man study of the CoStar Paclitaxel-Eluting Coronary Stent System evaluating three dose release formulations of paclitaxel in a bioresorbable polymer matrix in the treatment of de novo coronary lesions. METHODS: The COSTAR I Trial was a prospective, multi-center registry enrolling 87 patients in four Indian centers for treatment of up to two de novo lesions = 25 mm in length in a reference vessel 2.5-3.5 mm in diameter. Three dose release formulations were studied: 30 microg eluted over 10 days bidirectionally (Group 1, n =10), 10 microg eluted over 30 days abluminally (Group 2, n=40) and 3 microg eluted over 30 days abluminally (Group 3, n = 37). RESULTS: Demographics and lesion characteristics were similar between the groups and treatment in all three groups included small caliber vessels (RVD 2.45 +/- 0.30 - 2.57 +/- 0.36 mm). The primary endpoint of in-stent late loss at four months was lowest in Group 2 (0.43 +/- 0.43 mm) compared to Group 1 and Group 3 (0.51 +/- 7 mn; 0.74 mm and 1.07 +/- 0.65 mm respectively). In-segment late loss followed similar trends, being lowest in Group 2 (0.24 +/- 0.39 mm) compared to Groups 1 and 3 (0.52 +/- 0.66 mm and 0.76 +/- 0.57 mm respectively). Group 2 demonstrated better angiographic out-comes at 12 months with in-stent late loss of 0.55 +/- 0.38 mm when compared to Groups 1 and 3 (0.90 +/- 0.76 mm and 0.74 +/- 0.55 mm respectively). Cumulative binary restenosis rates at twelve months were 1.9%, 35.7% and 39.1% in Groups 2, 1 and 3 respectively. Clinical outcomes trended similarly with cumulative MACE rates at twelve months being lowest at 7.5% in Group 2 as compared to 20% in Group 1 and 21.6% in Group 3 respectively. CONCLUSIONS: In this first-in-man feasibility trial, angiographic and clinical results seen with the extended release formulation at a higher dose (10 microg/30 days) demonstrate the feasibility of the CoStar stent platform in the treatment of native coronary lesions. It also demonstrates the importance of drug dose and release kinetics. PMID- 19122252 TI - Value of CRP in coronary risk determination. AB - Initial studies showed that C-reactive protein (CRP) may have a role in the genesis of atherosclerotic lesions and as a novel biomarkers of patients at risk of developing coronary heart disease (CHD) events. Most of these studies had methodologic limitations, and recent data contradict these findings and suggest that inflammatory markers, such as CRP, have limited usefulness in the prediction of CHD events over and above conventional risk factors. Its low predictive value creates difficulty in interpreting the CRP data in an individual patient. Current scientific literature does not support measurement of CRP in individual patients in clinical medicine. In this review, we critically appraise role of CRP in cardiovascular medicine. PMID- 19122253 TI - Familial arterial tortuosity syndrome. PMID- 19122254 TI - Long term cardiac abnormality after single high dose exposure to sulfur mustard? PMID- 19122255 TI - Cardiac resynchronization therapy in hemodynamically unstable heart failure patients. PMID- 19122256 TI - Utility of micro-coils and glue in coronary artery perforation during balloon angioplasty. PMID- 19122257 TI - Longitudinal mitral annular early diastolic velocities in differentiating effusive constrictive pericarditis from restrictive cardiomyopathy: a case with extreme and straddling variations in regional velocities. PMID- 19122258 TI - Percutaneous transluminal catheter-based coil embolization of septal artery as a treatment for hypetrophic obstructive cardiomyopathy. PMID- 19122259 TI - Implanting a left ventricular pacemaker lead. PMID- 19122260 TI - [New development in bisphosphonate treatment. Fracture prevention by bisphosphonates]. AB - Alendronate and risedronate have been the standard regimens in the pharmacological intervention of osteoporosis based on their accumulated evidences of fracture prevention. At the start of bisphosphonate therapy, fracture risk assessment should be conducted for each patient, and the indication to use other drugs should be paid attention according to the patients' situation. At present, combination therapy with other drugs needs clinical decision at each clinical setting. A large scale prospective study is being conducted in Japan to answer the question whether the combination therapy with alendroate and alfacaldidol is more effective than alendronate alone to prevent fractures. PMID- 19122261 TI - [New development in bisphosphonate treatment. Effect of bisphosphonate on structural property]. AB - Bisphosphonate is highly potent antiresorptive agent with an effect to preserve bone structure. Structural property is evaluated using bone imaging techniques. Hip Structure Analysis (HSA) program based on 2D hip DXA data, estimates 3D geometrical properties such as cross-sectional area (CSA) , subperiosteal width and average cortical thickness. From these geometrical parameters, biomechanical indices can be derived. HSA shows BP has an effect to prevent of endocortical resorption and to increase cortical thickness, and finally to increase biomechanical strength. Micro-CT visualizes and measures 3D trabecular microarchitecture, which shows that BP prevents osteoporotic microstructural deterioration, such as trabecular disconnectivity, decrease of trabecular number, and transformation from plate-like structure to rod-like structure. These changes are considered to be related to increase biomechanical strength. PMID- 19122262 TI - [New development in bisphosphonate treatment. Characteristics and effectiveness of intermittent bisphophonates]. AB - Despite widely accepted efficacy of bisphophonates for osteoporotic fracture, patients treatment compliance has been low, especially, with daily regimen, in part because of the inconvenience associated with special dosing requirements of oral bisphophonates. Intermittent bisphophonates, oral and intravenous, have been introduced to solve the problem. In Japan, at present, only weekly alendronate and risedronate, both oral, have been approved for osteoporosis. In U. S. A. and other countries, orally, monthly risedoronate and ibandronate, intravenously, quarterly ibandronate and yearly zoledronate have been approved for osteoporosis. Their clinical efficacy will be reviewed. PMID- 19122263 TI - [New development in bisphosphonate treatment. When and how long should patients take bisphosphonates for osteoporosis?]. AB - Bisphosphonates such as alendronate and risedronate effectively reduce hip fracture risk, and thus are suitable for the elderly people in whom this fracture is most likely to happen. Bisphophonates should be administered when patients meet the guideline made by Japanese bone society, which picks up prevalent fractures, low bone mineral density, and established risk factors as features for starting osteoporosis treatment. Long-term alendronate administration may inhibit normal repair of microdamage arising from severe suppression of bone turnover (SSBT), which, in turn, results in accumulation of microdamage. This process would lead to brittle bone and the occurrence of unexpected stress fractures, characteristically at the subtrochanter of femur. A large-scale study suggests that for more women, discontinuation of alendronate after 5 years for up to 5 more years does not significantly increase fracture risk. Thus, alendronate treatment might be stopped for a while after 5 years to prevent SSBT and subsequent stress fractures. PMID- 19122264 TI - [New development in bisphosphonate treatment. Bisphosphonates for the treatment of glucocorticoid-induced osteoporosis]. AB - Only bisphosphonates were shown to decrease the risk of vertebral fractures in patients taking glucocorticoids. Relative risk at 1 year treatment with bisphosphonates for incident vertebral fractures and non-vertebral fractures were 0.46 (95% confidence interval : 0.28 - 0.77) and 0.77 (95% confidence interval : 0.39 - 1.51) , respectively. Two years extension of the alendronate trial showed that relative risk for incident vertebral fractures was 0.10 (95% confidence interval : 0.01 - 0.90) . PMID- 19122265 TI - [New development in bisphosphonate treatment. Treatment with bisphosphonates for cancer-induced bone disease]. AB - Bone metastases and multiple myeloma cause debilitating clinical symptoms including intractable bone pain, disabling multiple fractures and hypercalcemia. Adjuvant endocrine therapies with aromatase inhibitors or androgen deprivation further accelerate bone loss and increase in fracture risk (cancer treatment induced bone loss) . Bisphosphonates appear to be a current mainstay for the treatment and prevention of the skeletal-related morbidity in cancers. However, osteonecrosis of the jaw emerges as a major concern associated with long-term bisphosphonate use. It is important to clarify the clinical impact of long-term bisphosphonate use and make beneficial use of this agent by eliminating underlying risk of its adverse effects. PMID- 19122266 TI - [New development in bisphosphonate treatment. Review of effect on bone metabolism by minodronic acid in primary osteoporosis]. AB - Minodronic acid hydrate is a novel bisphosphonate with stronger anti-resorptive effects, compared to the other existing bisphosphonates. We performed additional analyses of phase III clinical trial and compared the effect of minodronic acid hydrate on bone turnover markers with those of alendronate in primary osteoporosis. The results showed that minodronic acid hydrate suppressed bone turnover early after treatment, and it also was effective in the treatment of patients with prevalent vertebral fracture, which is a high risk for subsequent fracture. PMID- 19122267 TI - [New development in bisphosphonate treatment. Review of the preventive effect of minodronic acid on fracture in Japanese patients with osteoporosis]. AB - Minodronic acid, a novel bisphosphonate, is one of the osteoporosis medications that has a potent inhibitive action on bone resorption. In this study, we performed additional analyses of the results on the already-reported phase III fracture study using minodronic acid, and reviewed the efficacy in elderly patients aged 75 years or older. The results show that minodronic acid suppressed the occurrence of vertebral fracture in patients aged 75 years or older at higher risk of fracture, in the same way as in patients aged less than 75 years. The phase III fracture study using minodronic acid was the first study that verified the preventive effect on vertebral fracture versus placebo in Japanese patients with osteoporosis. The results of the study compared favorably with placebo controlled large-scale clinical studies using other agents that were conducted overseas. PMID- 19122268 TI - [New development in bisphosphonate treatment. Clinical significance of combinatorial therapies with bisphosphonates and parathyroid hormone for osteoporosis]. AB - Bisphosphonates are the first line therapy to prevent osteoporotic fractures in daily practice. However, their preventive effect for fracture is not always satisfactory. In particular, more effective therapeutic tools are needed for patients who report osteoporotic fractures during treatment with bisphosphonates. To this end, treatment with parathyroid hormone has demonstrated potent bone anabolic effects and efficacy to prevent fractures, and physicians in Japan are looking forward to government approval to its clinical application. However, since, due to its characteristics, such as daily injection and expensive costs, parathyroid hormone is not the best candidate as the first line therapy for osteoporosis, we need to investigate how to use parathyroid hormone in combination with other drugs for osteoporosis, such as bisphosphonates. PMID- 19122269 TI - [New development in bisphosphonate treatment. Bisphosphonates and gastrointestinal adverse effects]. AB - Endoscopic examination suggested gastric mucosal damage by NSAID is more serious than that of bisphosphonates. In addition, the incidence of GI-adverse effects (AEs) is reported to be insignificant, if the incidence is adjusted by history of peptic ulcer, drugs for comorbidity, sex, age and/or ways of evaluation of the AEs. The anxiety for taking medication under empty stomach may contribute to the increase of GI-AEs. Some consideration must be taken for patients with past history of peptic ulcer, and/or drug user such as NSAIDs. PMID- 19122270 TI - [New development in bisphosphonate treatment. Bisphosphonate therapy and osteonecrosis of the jaws]. AB - Bisphosphonates (BP) have clinically been used as a highly effective drug in the treatment of hypercalcemia of malignancy, multiple myeloma, skeletal events associated with metastatic breast cancer and prostate cancer, and osteoporosis. Despite these benefits, however, the emergence of BP-related osteonecrosis of the jaws (BRONJ) becomes a growing and significant problem in a subset of patients receiving these drugs, especially intravenous preparations. It has also been reported in the patients receiving oral BP, although the incidence is extremely low. Most of BRONJ cases occur after dental treatments such as tooth extraction, periodontal surgery, and dental implants, and are refractory to conventional treatment modalities such as debridement, antibiotics and hyperbaric oxygen therapy. As compared to EU and USA, the number of BRONJ case is still small in Japan, but it is exactly increasing year by year. The ratio of the number of BRONJ in patients receiving oral BP to that in patients receiving intravenous BP is higher in Japan than in EU and USA, speculating due to the difference of time of approval. In this communication, the practical guidelines for prevention, diagnosis and treatment of BRONJ recently released from USA and Canada were introduced. Although no effective therapy for BRONJ is established yet, the importance of oral hygiene, patient education and treatments suitable for clinical stage is emphasized. In addition, it is considered that the survey of incidence of BRONJ in Japan and the preparation of Japanese guideline are urgent need. PMID- 19122271 TI - [Rickets]. AB - This review mainly describes on childhood vitamin D deficiency and x-linked hypophosphatemic rickets. Though nutritional state has improved dramatically, 25 (OH) D level, which is a good indicator of vitamin D status, is marginal especially in winter, and vitamin D deficiency is not rare in Japan. The PTH/Vitamin D axis does not account for the entire picture of x-linked hypophosphatemic rickets, and a new bone (osteocyte) -renal metabolic milieu has emerged and loss of PHEX, mostly expressed in osteocytes, is proposed to result in inappropriate processing of MEPE and consequent reduction in bone mineralization and an increase in circulating FGF23 to give rise to phosphaturia and hypophosphatemia. PMID- 19122272 TI - Current application and technology of functional multineuron calcium imaging. AB - Here we describe recent applications and technical advancements of functional multineuron calcium imaging (fMCI), which monitors the firing activity of more than a thousand neurons through their somatic Ca(2+) signals. fMCI is used for analysis of various neural circuits under normal and pathological conditions. In vitro fMCI is made more sophisticated by using multipoint illumination and scanning technology with spinning-disk and low-laser-intensity imaging, electron multiplying charge-coupled device cameras, etc. In vivo fMCI is still developing. We review optical technologies for fast scanning imaging, deep tissue imaging, and recording from moving animals. PMID- 19122273 TI - Effect of bone morphogenetic protein-2 (BMP-2) or troglitazone, as an inducer of osteogenic cells or adipocytes, on differentiation of a bone marrow mesenchymal progenitor cell line established from temperature-sensitive (ts) simian virus (SV) 40 T-antigen gene transgenic mice. AB - TBR31-2 is one of the bone marrow stromal cell lines. Differentiation toward osteogenic cells and calcification was observed when TBR31-2 cells were cultured for 4 weeks. Bone morphogenetic protein-2 (BMP-2) stimulated alkaline phosphatase (ALP) activity in a dose- and time-dependent manner. On the other hand, troglitazone increased oil droplet accumulation in a dose-dependent manner. In the presence of BMP-2, an increase of expression in osteogenic cell differentiation marker genes and a decrease of expression in adipocyte differentiation marker genes were observed with the exception of the induced expression of peroxisome proliferator-activated receptor gamma (PPARgamma), however, troglitazone, a ligand of PPARgamma treatment exhibited the opposite tendency. Interestingly, treatment with both BMP-2 and troglitazone resulted in a decrease of ALP activity and an increase of oil droplet accumulation. Reverse tanscription-polymerase chain reaction (RT-PCR) analysis also indicated that osteogenic differentiation markers decreased and that adipocyte differentiation markers increased. Thus, when the cells were cultured with BMP-2, osteogenic differentiation was enhanced while the expression of PPARgamma was maintained, and the addition of troglitazone caused a significant number of differentiated cells into adipocytes. These findings indicate that BMP-2 enhanced osteogenic differentiation and the expression of adipogenic transcription factor (PPARgamma) followed by osteogenic differentiation without activation of PPARgamma by its ligand. PMID- 19122274 TI - Effects of ATP on the intracellular calcium level in the osteoblastic TBR31-2 cell line. AB - We investigated the effects of extracellular ATP on TBR31-2 cells established from the bone marrow of transgenic mice harboring the temperature-sensitive simian virus (SV) 40 T-antigen gene. These cells showed the capacity to differentiate toward osteoblasts and could be enhanced by bone morphogenetic protein (BMP)-2, an inducer of osteoblasts. The intracellular calcium ion level ([Ca(2+)](i)) in differentiating TBR31-2 cells was measured by fluorescence confocal microscopic imaging using the Ca(2+)-sensitive probe, Calcium Green 1/AM. P2 receptor agonists, such as ATP (1 microM), uridine 5'-triphosphate (1 microM), and ADP (1 microM), significantly increased the [Ca(2+)](i) of TBR31-2 cells in 2-d and 5-d cultures, but a potent P2X receptor agonist, alpha,beta methylene ATP (10 microM), did not increase [Ca(2+)](i). The increase in [Ca(2+)](i) induced by ATP in the 2-d culture tended to be higher than in the 5-d culture. The increase in [Ca(2+)](i) of both cultures was inhibited by pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid, a P2 receptor antagonist. However, in an external Ca(2+)-free condition ATP-induced increase in [Ca(2+)](i) was unchanged at either stage. U73122, phospholipase C inhibitor and Thapsigargin, a calcium-pump inhibitor, significantly inhibited the increase in [Ca(2+)](i) at both stages. Reverse transcription-polymerase chain reaction analysis showed that the expression of P2Y receptor mRNA was higher in the 2-d culture than in the 5-d culture. These results indicate that ATP induces the increase in [Ca(2+)](i) from the calcium store through activating P2Y receptors in TBR31-2 cells and that the 2-d culture can respond to ATP more than the 5-d culture due to the higher expression of P2Y receptors. This suggests that the physiological role of ATP in osteoblasts is altered during differentiation. PMID- 19122275 TI - Development of SCAR markers for the discrimination of three species of medicinal plants, Angelica decursiva (Peucedanum decursivum), Peucedanum praeruptorum and Anthricus sylvestris, based on the internal transcribed spacer (ITS) sequence and random amplified polymorphic DNA (RAPD). AB - Angelicae decursivae radix ('Jeonho' in Korean) is prescribed as the root of Angelica decursiva (= Peucedanum decursivum) and Peucedanum praeruptorum in Korean pharmacopoeia. However, Anthricus sylvestris has been usually distributed on the market because it is identical to the Korean plant name 'Jeonho'. Furthermore, due to the morphological similarity of the aerial parts and herbal medicines, the correct identification of these roots is difficult. Therefore, to develop a reliable method for discriminating among A. decursiva (= P. decursivum), P. praeruptorum and A. sylvestris, we applied the tools of molecular genetics, such as the analysis of ribosomal DNA internal transcribed spacer (rDNA ITS) and random amplified polymorphic DNA (RAPD). In the comparison of rDNA-ITS sequences, we found a specific primer region for the identification of A. sylvestris among three varieties of the herb that produced a 273 bp strand of DNA specific to A. sylvestris. As the result of RAPD analysis, we developed one sequence characterized amplified region (SCAR) marker for A. decursiva and P. praeruptorum that amplified a 363 bp DNA fragment specific to both A. decursiva and P. praeruptorum and two markers for P. praeruptorum that amplified 145 bp and 305 bp DNA fragments specific to P. praeruptorum. Furthermore, we established the SCAR markers for the simultaneous discrimination of the three species by applying a multiplex-polymerase chain reaction (PCR) with a combination of primers. This method of discrimination would be useful in preventing the distribution of adulterates because it can identify each herb and distinguish it from inauthentic substitutions. PMID- 19122276 TI - Functional analysis of aldehyde oxidase using expressed chimeric enzyme between monkey and rat. AB - Aldehyde oxidase (AO) is a homodimer with a subunit molecular mass of approximately 150 kDa. Each subunit consists of about 20 kDa 2Fe-2S cluster domain storing reducing equivalents, about 40 kDa flavine adenine dinucleotide (FAD) domain and about 85 kDa molybdenum cofactor (MoCo) domain containing a substrate binding site. In order to clarify the properties of each domain, especially substrate binding domain, chimeric cDNAs were constructed by mutual exchange of 2Fe-2S/FAD and MoCo domains between monkey and rat. Chimeric monkey/rat AO was referred to one with monkey type 2Fe-2S/FAD domains and a rat type MoCo domain. Rat/monkey AO was vice versa. AO-catalyzed 2-oxidation activities of (S)-RS-8359 were measured using the expressed enzyme in Escherichia coli. Substrate inhibition was seen in rat AO and chimeric monkey/rat AO, but not in monkey AO and chimeric rat/monkey AO, suggesting that the phenomenon might be dependent on the natures of MoCo domain of rat. A biphasic Eadie-Hofstee profile was observed in monkey AO and chimeric rat/monkey AO, but not rat AO and chimeric monkey/rat AO, indicating that the biphasic profile might be related to the properties of MoCo domain of monkey. Two-fold greater V(max) values were observed in monkey AO than in chimeric rat/monkey AO, and in chimeric monkey/rat AO than in rat AO, suggesting that monkey has the more effective electron transfer system than rat. Thus, the use of chimeric enzymes revealed that 2Fe-2S/FAD and MoCo domains affect the velocity and the quantitative profiles of AO-catalyzed (S)-RS 8359 2-oxidation, respectively. PMID- 19122277 TI - The effect of plagiochin E alone and in combination with fluconazole on the ergosterol biosynthesis of Candida albicans. AB - Plagiochin E (PLE), a macrocyclic bis(bibenzyl) isolated from the liverwort Marchantia polymorpha, has been reported to have antifungal activity and resistance reversal effects on Candida albicans. In order to understand the underlying mechanisms, we studied the effects of PLE alone and in combination with fluconazole (FLC) on the ergosterol biosynthetic pathway against both FLC sensitive and FLC-resistant strains by analyzing the sterol content and the ergosterol pathway gene (ERG) expression. Relative quantitative analysis of different ergosterol precursors was carried out by employing the hyphenated technique of gas chromatography-high resolution mass spectrometry (GC-HR-MS). We observed that for FLC-resistant strain PLE itself can cause the accumulation of lanosterol and the decrease of 14alpha-methylfecosterol. When it combined with FLC, a significant decrease was observed in ergosterol formation and corresponding accumulation of 14alpha-methylated sterols was also found. Real time reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that the transcription level of ERG11 was decreased in FLC-resistant strain when exposed to PLE alone or PLE plus FLC. These results suggest that PLE potentiates FLC antifungal activity by interfering with the FLC-targeted ergosterol biosynthesis pathway. PMID- 19122278 TI - Anti-infectious activity of tryptophan metabolites in the L-tryptophan-L kynurenine pathway. AB - To study the anti-infectious effect of a vascular allograft, the antimicrobial activity of tryptophan metabolites mediated by indoleamine 2,3-dioxygenase was determined. The growth of methicillin-resistant Staphylococcus aureus (MRSA) over 10 h in extracts from post-transplantation vascular allograft was significantly slower than that of extracts from non-transplantation vascular allograft regardless of the presence of tryptophan (p<0.05). When the antimicrobial activity of the tryptophan metabolites in the L-tryptophan-L-kynurenine pathway was examined, 3-hydroxy-DL-kynurenine and alpha-picolinic acid had strong antibacterial activity against MRSA, S. epidermidis, Escherichia coli, and multidrug-resistant Pseudomonas aeruginosa, although antimicrobial activities of anthranilic acid, 3-hydroxyanthranilic acid, and quinolinic acid against them were low. The results showed that, of the tested tryptophan metabolites, 3 hydroxy-DL-kynurenine and alpha-picolinic acid contributed to the anti-infectious effects of the allograft by inhibiting of the growth of microorganisms. PMID- 19122279 TI - Pyridone 6, a pan-Janus-activated kinase inhibitor, suppresses osteoclast formation and bone resorption through down-regulation of receptor activator of nuclear factor-kappaB (NF-kappaB) ligand (RANKL)-induced c-Fos and nuclear factor of activated T cells (NFAT) c1 expression. AB - It has been reported that Janus tyrosine kinase (JAK)-dependent signaling pathways play a critical role in the pathogenesis of numerous malignancies and immune reactions, and inhibition of JAK has been implicated in cell growth inhibition. The role which JAK has on osteoclast differentiation and anti-bone resorptive activity is not well understood. In this study, we investigated the effects of a pan-JAK inhibitor, pyridone 6, on osteoclast differentiation and bone-resorption in vitro and ex vivo. Pyridone 6 inhibited osteoclast differentiation in mouse bone marrow macrophage (BMM) cultures stimulated by the receptor activator of nuclear factor-kappaB (NF-kappaB) ligand (RANKL) and co cultures of bone marrow cells and osteoblasts. Pyridone 6 suppressed the expression of c-Fos and nuclear factor of activated T cells (NFAT) c1 in BMMs. It also inhibited the bone resorptive activity of mature osteoclasts that was accompanied by disruption of actin rings. Pyridone 6 also suppressed I-kappaB degradation and extracellular signal-regulated kinase (ERK) in mature osteoclasts, suggesting that these are the key molecules that pyridone 6 targets in the inhibition of osteoclast function. These results demonstrate inhibition of JAK may be useful for the treatment of bone-resorptive diseases, such as osteoporosis. PMID- 19122280 TI - Effects of histamine H1-antagonists on sleep-awake state in rats placed on a grid suspended over water or on sawdust. AB - The present study was performed to investigate the effects of histamine H(1) antagonists on the sleep-awake state in rats placed on a grid suspended over water in comparison with rats placed on sawdust. When rats were placed on the grid suspended over water, significant increases in the awake time and decreases in non-rapid eye movement (non-REM) sleep time were observed compared with in rats on sawdust, even when measured hourly for 6 h. Diphenhydramine, chlorpheniramine and promethazine caused a significant decrease in the awake time and increase in non-REM sleep time in rats placed on the grid suspended over water for 1-2 h and/or 2-3 h after administration. On the other hand, in rats placed on sawdust, no significant differences were observed in the awake time and non-REM sleep time with diphenhydramine and chlorpheniramine compared with the control. Different from these two drugs, promethazine caused a significant decrease in the awake time and increase in non-REM sleep time 1-2 h and 2-3 h after administration even when rats were placed on sawdust at a relatively high dose. These results clearly indicate that histamine H(1)-antagonists had potent effects on decreasing the awake time and increasing non-REM sleep time under the conditions of an activated histaminergic system. PMID- 19122281 TI - Neuroprotective effects of (-)-epigallocatechin-3-gallate on aging mice induced by D-galactose. AB - The aim of the present study was to investigate the protective effects of (-) epigallocatechin-3-gallate (EGCG), the main polyphenolic constituent of green tea, in aging mice induced by D-galactose (D-gal). The aging mice model was induced by subcutaneous (s.c.) injection of D-gal (150 mg/kg) once daily for 6 weeks. EGCG (2 mg/kg or 6 mg/kg) was administered intragastrically (i.g.) once daily for 4 weeks after 2-week D-gal injection. The water maze test was used to evaluate the learning and memory function of mice. The activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) and the contents of malondialdehyde (MDA) in the hippocampus were measured using different biochemical kits to estimate the changes in the antioxidative ability of mice. TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining method was used to detect neuronal apoptosis, and the activation and expression of proapoptotic protein caspase-3 in the hippocampus were observed and analyzed using immunohistochemical staining and the Western blot method to evaluate apoptosis in the brain. The results indicated that subcutaneous injection of D-gal induced learning and memory impairment in mice, decreased T-SOD and GSH-Px activities, increased MDA contents in the hippocampus, and increased the cell apoptosis index and cleaved caspase-3 protein expression in the hippocampus. Oral administration of EGCG (2 mg/kg or 6 mg/kg) for 4 weeks significantly improved the cognitive deficits in mice and elevated T-SOD and GSH-Px activities, decreased MDA contents in the hippocampus, and reduced the cell apoptosis index and expression of cleaved caspase-3 in the mouse hippocampus. The results suggest that EGCG has potent neuroprotective effects on aging mice induced by D-gal through antioxidative and antiapoptotic mechanisms, indicating that EGCG is worthy of further study in aging. PMID- 19122282 TI - Protective effect of quercetin against gentamicin-induced nephrotoxicity in rats. AB - Gentamicin (GM) is an antibiotic widely used in treating severe gram-negative infections. However, its clinical use is limited by its nephrotoxicity. Several lines of evidence indicate that free radicals are important mediators of gentamicin nephrotoxicity. Therefore, the aim of this work was to investigate the possible protective effect of the flavonoid quercetin, an antioxidant, on gentamicin-induced nephrotoxicity. For this purpose, rats were divided into four groups. First group served as a control and injected with the normal saline, second group was injected with quercetin (50 mg/kg/d, per os) for 7 d, third group was injected with gentamicin (80 mg/kg/d, intraperitoneally) for 7 d and the fourth group of animals was injected with quercetin plus gentamicin simultaneously for 7 d. Total protein levels were estimated in 24-h urine samples to assess kidney dysfunction. The rats were sacrificed on the seventh day and kidneys were collected for histopathological studies. Blood urea nitrogen (BUN) and creatinine levels were measured in the blood. Moreover, glutathione (GSH), lipid peroxide (TBARS) levels, superoxide dismutase (SOD) and catalase (CAT) activities were determined in renal tissues. GM-treated rats showed early kidney dysfunction as urinary total protein, BUN and serum creatinine levels were significantly increased. The significant decrease in GSH levels, SOD, CAT activities and increase in TBARS levels, indicated that GM-induced nephrotoxicity was mediated through oxidative stress reactions. Histopathological examination of GM-treated rats revealed degenerative changes in glomeruli and tubules. On the other hand, simultaneous administration of quercetin plus gentamicin protected kidney tissues against nephrotoxic effects of gentamicin as evidenced from amelioration of histopathological changes and normalization of kidney biochemical parameters. PMID- 19122283 TI - In vivo and in vitro antiviral activities of calycosin-7-O-beta-D-glucopyranoside against coxsackie virus B3. AB - Calycosin-7-O-beta-D-glucopyranoside (CCGR) is the main isoflavonoid compound isolated from Astragalus membranaceus var. Mongholicu (BGE.) HSIAO, a Chinese herb medicine traditionally used to treat viral myocarditis. In this study, its antiviral activities against coxsackie virus B3 (CVB3) causing myocarditis were investigated. In vitro assay showed that CCGR displayed a low cytotoxicity and effectively inhibited CVB3-mediated cytopathic effects on Vero cells with an IC(50) value of 25 microg/ml. In an acute myocarditis murine model, treatment with 24 mg/kg CCGR for 14 d significantly improved the survival rate of mice infected with CVB3, alleviated pathological damages of cardiac muscles in the myocarditis mice, reduced the virus titers in the heart, decreased heart indexes and improved left ventricular function. These results showed that CCGR exerted significant antiviral activities against CVB3 both in vitro and in vivo, and identified CCGR as one of active ingredients in Astragalus membranaceus for the treatment of viral myocarditis. PMID- 19122284 TI - Involvement of P-glycoprotein and multidrug resistance associated protein 1 on the transepithelial transport of a mercaptoacetamide-based histone-deacetylase inhibitor in Caco-2 cells. AB - Oral bioavailability is one of the important criteria for development of a drug lead candidate. In this study, the absorptive characteristics and the efflux mechanism of a mercaptoacetamide-based histone deacetyalse (HDAC) inhibitor, coded as W2, were investigated using Caco-2 cells. The transport of W2 was asymmetric as indicated by 1.85 fold higher basolateral to apical (BL to AP) than apical to basolateral (AP to BL) flux. Such asymmetry was associated with multidrug resistance associated protein 1 (MRP1) and P-glycoprotein (P-gp), as evidenced by specific inhibition of these proteins. In the presence of verapamil and cyclosporin A, potent inhibitors of P-gp, the apparent permeability ratio (P(app) BL to AP/P(app) AP to BL) of W2 was decreased from 1.85 to 0.73 and 1.03, respectively, and the absorption from apical to basolateral side was enhanced from 13.3+/-0.2x10(-6) cm/s to 17.3+/-0.12x10(-6) cm/s and 19+/-0.3x10(-6) cm/s, respectively. Upon addition of quinidine, a mixed P-gp and MRP1 inhibitor, the permeation of W2 from the apical side was significantly increased (P(app) 17.1+/ 0.32x10(-6) cm/s) while the efflux was inhibited (P(app) 21.3+/-0.19x10(-6) cm/s). Furthermore, the influence of the MRP1 inhibitors, indomethacin and N benzyl-indomethacin (NBI) was evaluated. NBI treatment attenuated the basolateral to apical flux of W2 (P(app) 20.3+/-0.1x10(-6) cm/s), whereas this effect was completely abrogated by indomethacin (P(app) 11+/-0.4x10(-6) cm/s). The results suggest that P-gp and MRP1 transporters are capable of mediating the efflux of W2 and might play a significant role in its oral absorption. PMID- 19122285 TI - Berberine exerts neuroprotective actions against in vitro ischemia-induced neuronal cell damage in organotypic hippocampal slice cultures: involvement of B cell lymphoma 2 phosphorylation suppression. AB - In this study we elucidated the effects of berberine, a major alkaloid component contained in medicinal herbs, such as Phellodendri Cortex and Coptidis Rhizoma, on ischemic neuronal damage in mouse organotypic hippocampal slice cultures (OHSCs) caused by oxygen and glucose deprivation (OGD) and N-methyl-D-aspartate (NMDA) -type glutamate receptor stimulation. Hippocampal slices obtained from 7-d old ICR mice were cultured for 10 d before the experiments. Ischemia-related damage was induced by OGD (5, 15, 45 min) or NMDA (10 microM) treatment, and was evaluated by measuring propidium iodide (PI) uptake. Levels of apoptotic marker proteins, B-cell lymphoma 2 (Bcl-2) and phosphorylated-Bcl-2 (p-Bcl-2), in the OHSCs were measured as indices of biochemical neuronal cell damage by Western blotting. Berberine (5, 25 microM) or the NMDA antagonist MK-801 (25 microM) was added to the medium 30 min before OGD or NMDA treatment. OGD time-dependently increased PI uptake of the OHSCs. Both berberine (5, 25 microM) and MK-801 (25 microM) significantly inhibited PI uptake at 24 h after 45-min OGD treatment and PI uptake in OHSCs exposed to NMDA for 24 h. OGD treatment also significantly increased the level of p-Bcl-2 but not that of Bcl-2 or glyceraldehyde-3 phosphate dehydrogenase (GAPDH) in OHSCs. Berberine (5-25 microM) significantly suppressed the OGD-induced increase of p-Bcl-2 level in OHSCs when tissue was exposed to the alkaloid prior to OGD or simultaneously with OGD. These findings suggest that berberine has protective effects against ischemic damage in mouse OHSCs and that the effects are at least partly mediated by suppression of Bcl-2 phosphorylation. PMID- 19122286 TI - Inhibitory effect of 2,4,2',4'-tetrahydroxy-3-(3-methyl-2-butenyl)-chalcone on tyrosinase activity and melanin biosynthesis. AB - 2,4,2',4'-Tetrahydroxy-3-(3-methyl-2-butenyl)-chalcone (TMBC), a naturally occurring compound from Morus nigra, modulated melanogenesis by inhibiting tyrosinase. TMBC inhibited the L-dopa oxidase activity of mushroom tyrosinase with an IC(50) value of 0.95+/-0.04 microM, which was more potent than kojic acid (IC(50)=24.88+/-1.13 microM), a well-known tyrosinase inhibitor. The kinetic studies of tyrosinase inhibition revealed that TMBC acts as a competitive inhibitor of mushroom tyrosinase with L-dopa as the substrate. Furthermore, TMBC effectively inhibited both cellular tyrosinase activity and melanin biosynthesis in B16 melanoma cells without significant cytotoxicity. The inhibitory effect of TMBC on melanogenesis was attributed to the direct inhibition of tyrosinase activity, rather than the suppression of tyrosinase gene expression. These results indicated that TMBC may be a new promising pigmentation-altering agent for cosmetic or therapeutic applications. PMID- 19122287 TI - 3-Oxoolean-12-en-27-oic acid isolated from Aceriphyllum rossii induces caspase-8 dependent apoptosis in human promyelocytic leukemia HL-60 cells. AB - In the present study, we investigated the effects of 3-oxoolean-12-en-27-oic acid (3-OA) isolated from the underground parts of Aceriphyllum rossii (Saxifragaceae) on the viability and apoptosis of HL-60 human promyelocytic leukemia cells, and the mechanisms underlying its action. 3-OA-treated HL-60 cells and HeLa human cervix adenocarcinoma cells displayed several apoptotic features, such as, DNA fragmentation, DNA laddering by agarose gel electrophoresis, and hypodiploid DNA contents by flow cytometry, and 3-OA also caused the activations of caspase-8, -9 and -3. Pretreatment with z-VAD-fmk (a broad-caspase inhibitor) almost completely suppressed 3-OA-induced DNA ladder formation and hypodiploid DNA contents, thereby implicating the caspase cascade in the apoptotic process. In addition, z IETD-fmk (a caspase-8 inhibitor) and z-DEVD-fmk (a caspase-3 inhibitor) also completely neutralized the apoptotic effect of 3-OA in HL-60 cells. Furthermore, 3-OA increased Fas-related protein contents and the mRNA expressions of Fas ligand (FasL), Fas, and Fas-associated death domain (FADD). Preincubation with anti-Fas or anti-FasL blocking antibodies completely prevented 3-OA-induced apoptosis. Taken together, these results suggest that 3-oxoolean-12-en-27-oic acid induces apoptosis by activating caspase-8 via FasL-stimulated death receptor signaling. PMID- 19122288 TI - A retrospective analysis of vancomycin pharmacokinetics in Japanese cancer and non-cancer patients based on routine trough monitoring data. AB - The pharmacokinetics of vancomycin was retrospectively examined based on trough concentrations obtained during routine therapeutic drug monitoring to examine possible pharmacokinetic differences between adult Japanese cancer and non-cancer patients with various degrees of renal function. A total of 231 data points from 65 cancer patients and 41 non-cancer patients were collected, and patients' background, vancomycin dose, and vancomycin clearance estimated by an empirical Bayesian method were summarized. Regarding the patients' characteristics and clinical laboratory test data, no clear differences were found between the two groups. The relationship between vancomycin clearance and creatinine clearance were similar between the groups, suggesting little effect of malignancy on vancomycin clearance. After the sub-group comparisons regarding fluid retention and cancer type, no clear differences were found in the vancomycin clearance versus creatinine clearance relationship. We conclude that the initial dose of vancomycin should not necessarily be adjusted for cancer patients. For individualized vancomycin-based therapy, dose adjustment at the appropriate time is important according to information from routine therapeutic drug monitoring and clinical laboratory tests, and to observations of the efficacy, nephrotoxicity, and other conditions in each patient. PMID- 19122289 TI - Inhibitory effects of herbal extracts on the activity of human sulfotransferase isoform sulfotransferase 1A3 (SULT1A3). AB - Sulfotransferase 1A3 (SULT1A3) is a phase II detoxifying enzyme of xenobiotics predominantly expressed in the intestinal epithelium. Recent increase in the use of herbal extracts as dietary supplements may lead to an increase in the possibility of dietary supplement-drug interactions. The purpose of the present study was to investigate the effects of 18 herbal extracts on SULT1A3 activity and the possibility of interaction between medicinal drugs and herbal extracts. We examined the inhibitory potencies of 18 herbal extracts on the sulfation of dopamine, a typical substrate of SULT1A3, and ritodrine, a beta(2) stimulant, by human recombinant SULT1A3. The sulfation of dopamine was inhibited by extracts of banaba, green tea, Rafuma, grape seed, peanut seed coat, gingko biloba leaf, St. John's wort, gymnema and milkthistle. The IC(50) values of these herbal extracts were lower than the putative gastrointestinal concentration when the recommended dose was ingested. On the other hand, chlorella extract and rutin showed no inhibitory effects and wheat, mulberry and siberian ginseng had IC(50) values exceedingly higher than the putative gastrointestinal concentration. The inhibitory profiles of herbal extracts for the sulfation of ritodrine were comparable to those for the sulfation of dopamine. In conclusion, the extracts of herbs such as banaba and green tea potently inhibited SULT1A3 activity. These extracts may increase the bioavailability of drugs whose bioavailabilities were limited by the function of SULT1A3 on the intestinal epithelium. PMID- 19122290 TI - Effects of panduratin A isolated from Kaempferia pandurata ROXB. on the expression of matrix metalloproteinase-9 by porphyromonas gingivalis supernatant induced KB cells. AB - Periodontitis is initiated by accumulation of microbial plaque and activation of gingival inflammation through overexpression of matrix metalloproteinases (MMPs), leading to tissue destruction. Natural MMP inhibitors may be developed as therapeutic agents against periodontitis. In this study, panduratin A, a natural bioactive compound isolated from Kaempferia pandurata ROXB., was used to test its in vitro inhibitory activity against MMP-9 secretion from Porphyromonas gingivalis supernatant-induced human oral epidermoid carcinoma KB cells. Gelatin zymography, Western blot and RT-PCR analyses were performed to evaluate MMP-9 expression. The gelatin zymograms revealed that the main gelatinase secreted by P. gingivalis supernatant-induced KB cells migrated at 92 kDa, representing MMP 9. MMP-9 protein and mRNA levels were significantly decreased after panduratin A treatment (p<0.05). In contrast, panduratin A had no effect on tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2 mRNA. Panduratin A also suppressed urokinase type plasminogen activator (uPA) mRNA expression. These results suggest that panduratin A could potentially prevent periodontal inflammation by decreasing the levels of MMP-9 protein and mRNA. PMID- 19122291 TI - Preventive effect of co-administration of water containing magnesium ion on indomethacin induced lesions of gastric mucosa in adjuvant-induced arthritis rat. AB - It is well known that nonsteroidal anti-inflammatory drugs (NSAIDs) have significant side effects, such as gastroenteropathy, and that rheumatoid arthritis patients taking NSAIDs are more susceptible to NSAIDs-induced gastric lesions as compared with patients with other diseases. We demonstrate the preventive effect of the co-administration of bittern water (BW, nigari-sui in Japanese), which enables the effective intake of Mg(2+), on the ulcerogenic response to indomethacin in adjuvant-induced arthritis (AA) rats. Four kinds of BW with different Mg(2+) contents; ranging from 10-200 mg/l Mg(2+) (BW-10, 25, 50, 200) were used in this study. Arthritis was induced by the injection of 50 microl of a suspension of 10 mg/ml heat-killed butyricum (Mycobacterium butyricum) in Bayol F oil into the plantar region of the right hind foot and tail of rats. Oral administration of indomethacin (40 mg/kg) caused hemorrhagic lesions in the gastric mucosa of AA rats at 14 d after adjuvant injection, and the lesion score of AA rats administered indomethacin was significantly higher than that of normal rats administered indomethacin. The expression of the mRNA for inducible nitric oxide synthase (iNOS) mRNA expression and the production of nitric oxide (NO) in the gastric mucosa of AA rats were also increased by the administration of indomethacin. The co-administration of BWs decreased the ulcerogenic response to indomethacin in AA rats. In addition, the administration of BW attenuated the increase in iNOS mRNA expression and NO production in AA rats receiving indomethacin. The oral administration of Mg(2+) to AA rats had a potent preventive effect on the ulcerogenic response to indomethacin in AA rats, probably due to an inhibition in the rise in iNOS and NO levels in the gastric mucosa. PMID- 19122292 TI - Validation of HPLC-UV methods for quantitatively determining landiolol and its major metabolite in human blood. AB - Landiolol is an ultra-short-acting beta(1)-adrenergic receptor blocking agent that is used for both perioperative and postoperative patients with tachycardia during general anesthesia. Validated HPLC-UV methods that quantitatively determine landiolol and its major metabolite (M-1) in human blood were reported for clinical research of landiolol. These analytes were recovered from the same blood sample using a multi-step extraction process and determined with two different HPLC conditions. These methods were validated over concentration ranges of 0.05 to 10 microg/ml for landiolol and 0.1 to 20 microg/ml for M-1 and were found to have acceptable accuracy, precision, linearity, and selectivity. These methods are useful to the characterize of blood pharmacokinetics of landiolol and M-1 for clinical research. PMID- 19122293 TI - Nonviral vectors with a biosurfactant MEL-A promote gene transfection into solid tumors in the mouse abdominal cavity. AB - Recently, we showed that a biosurfactatnt 4-O-[(4',6'-di-O-acethyl-2',3'-di-O alkanoyl)-beta-D-mannopyranosyl] meso-erythritol A (MEL-A) greatly increased the efficiency of gene transfection mediated by cationic liposomes in vitro. We then studied whether the high transfection efficiency of these liposomes is maintained in vivo for tumor cells in the mouse abdominal cavity. When a complex of the liposomes and plasmid DNA was injected intraperitoneally into C57BL/6J mice bearing B16/BL6 tumors, we found that the biosurfactant significantly increased liposome-mediated gene transfection to the mouse tumor cells. The transfection efficiency of the plasmids into the solid tumors by the cationic liposomes of cholesteryl-3beta-carboxyamidoethylene-N-hydroxyethylamine (OH-Chol) with MEL-A increased by about 100-fold compared with that by the cationic liposomes of DC Chol (commercially available) without MEL-A. The results suggest that nonviral vectors with MEL-A are very useful for gene transfection in vivo. PMID- 19122294 TI - Effect of propranolol on hyphae formation signal in Candida albicans. AB - Candida albicans (C. albicans) is known as an opportunistic pathogen that changes from a yeast form to a hyphae form in response to various outside environmental signals. The addition of propranolol inhibited hyphae formation of C. albicans. Propranolol inhibited the expression of agglutinin like sequence 3 (ALS3) and ALS8mRNA, which are regulated by the cAMP-EFG1 pathway in C. albicans. Propranolol did not affect the expression of CST20, HST7 or CPH1mRNA, which are components of the mitogen-activated protein (MAP) kinase cascade in C. albicans. The expression of CYR1mRNA, which encodes adenylate cyclase of C. albicans, was not affected by propranolol. These findings indicated that the interruption of hyphae formation by propranolol is caused by inhibition of the cAMP-EFG1 pathway, but not effects on the MAP kinase cascade. PMID- 19122295 TI - Anti-hepatitis B virus activities of astragaloside IV isolated from radix Astragali. AB - Total ethanol extract and saponins from Chinese herb radix Astragali (huangqi) have been previously shown to possess anti-hepatitis B virus (HBV) activities in vitro. To identify the active ingredients, we isolated a triterpenoid saponin that was determined to be astragaloside IV. In the human HBV-transfected liver cell line HepG(2) 2.2.15, astragaloside IV effectively suppressed secretion of HBV antigens with inhibition rates of 23.6% for the secretion of Hepatitis B surface antigen (HBsAg) and 22.9% for that of Hepatitis B e antigen (HBeAg) at 100 microg/ml after 9 d of treatment. The inhibitory activity of astragaloside IV on secretion of HBV antigens is more potent than that of 3TC without significant cytotoxicity. In duck hepatitis B virus (DHBV)-infected ducklings, astragaloside IV caused 64.0% inhibition at 120 mg/kg, 49.6% inhibition at 40 mg/kg, and 41.7% inhibition at 10 mg/kg to serum DHBVs after 10 d of treatment, and also reduced serum DHBV DNA levels. Together, our results demonstrate that astragaloside IV possesses potent anti-HBV activity. PMID- 19122296 TI - Actinomycin D identified as an inhibitor of discoidin domain receptor 2 interaction with collagen through an insect cell based screening of a drug compound library. AB - Discoidin domain receptors belong to the cell surface receptor tyrosine kinase family and recognize collagens for their activating ligands. They have been implicated for cell growth and migration and their elevated expressions were observed in various human cancers. When we expressed human Discoidin domain receptor 2 (DDR2) in insect cells, the protein was targeted properly into the cell membrane and this could enforce the cells to adhere on culture plate coated with type I collagen. By taking advantage of this, we established a novel insect cell based screening protocol to identify chemicals which inhibit the interaction between DDR2 and collagen. We screened a drug-compound library to select an anti cancer drug, actinomycin D, as the inhibitory compound. Actinomycin D prevented the activation of DDR2 by type I collagen in human embryonic kidney 293 cells with an IC(50) value of 9 microM, while it did not interfere with the activation of other receptor tyrosine kinases by their ligands. In conclusion we identified a new biological function of actinomycin D and the insect cell based method provides a useful protocol for screening inhibitors against the association of DDR2 with collagen. PMID- 19122297 TI - Identification of Cistanche species by chemical and inter-simple sequence repeat fingerprinting. AB - Herba Cistanches is a common traditional Chinese medicine that has been used to reinforce the vital function of kidney and induce laxation for more than two thousands years. Four Cistanche species were found as Herba Cistanches in China herbal markets, including C. deserticola, C. tubulosa, C. salsa and C. sinensis. Phenylethanoid glycosides, particularly echinacoside and acteoside, are considered as active ingredients in Cistanche species. The contents of these compounds showed variation in different species and geographical sources. Standard chemical fingerprints were generated from each of four Cistanche species, which could be identification markers. In genetic analysis of Cistanche species, ninety-four inter-simple sequence repeat (ISSR) primers were used for polymerase chain reaction (PCR) amplification, and of which eight primers were found to be sufficient to distinguish different Cistanche species. As a result, the chemical fingerprint combined with the genetic fingerprint for distinction of Cistanche species could serve as markers for quality control of Herba Cistanches. PMID- 19122298 TI - Bioactive constituents from Chinese natural medicines. XXXIII. Inhibitors from the seeds of Psoralea corylifolia on production of nitric oxide in lipopolysaccharide-activated macrophages. AB - The methanolic extract from the seeds of Psoralea corylifolia was found to inhibit production of nitric oxide (NO) in lipopolysaccharide-activated mouse peritoneal macrophages. Among the isolated compounds, bavachinin (IC(50)=26 microM), isobavachalcone (17 microM), neobavaisoflavone (ca. 29 microM), corylifol A (ca. 21 microM), and psoralidin (ca. 23 microM) significantly inhibited the accumulation of nitrite (NO(2)(-)) as a marker of production of NO. Bakuchiol, which is mainly contained in the extract, also showed weak activity at 10 microM, but cytotoxic effects were observed more than 30 microM. PMID- 19122299 TI - Low dose estrogen supplementation reduces mortality of mice in estrogen-dependent human tumor xenograft model. AB - Breast cancer is one of the most frequent female cancers in the Western world. Perturbation of estrogen levels by hormone replacement therapy or pregnancy is associated with a variety of diseases, including breast cancer. Estrogen supplementation is required to establish appropriate animal models for estrogen related diseases. In this report, we demonstrated that supplementation with high doses of 17beta-estradiol results in deaths in estrogen-dependent MCF-7 tumor xenograft model. Renal damage and bladder stone formation was implicated as a major cause of death. The mortality rate was significantly reduced when mice received a low dose of 17beta-estradiol. We also confirmed that low dose of 17beta-estradiol supplementation can support the growth of tumors in MCF-7 tumor xenograft model. These results suggest that low dose estrogen supplementation may be more appropriate in estrogen-dependent tumor xenograft models. PMID- 19122300 TI - Improvement of diabetic complication by hydrangea dulcis folium in streptozotocin induced diabetic rats. AB - The improvement of diabetic complications such as lipid lowering and anti oxidative potential of Hydrangea Dulcis Folium (HDF) was studied in streptozotocin-induced diabetic rats. Male Sprague-Dawley rats were divided into 3 groups after induction of streptozotocin (STZ)-diabetes: normal control; diabetic control; diabetic-HDF supplement (hot water extract 40 g/kg diet); and fed experimental diets for 3 weeks. Serum glucose and insulin concentrations, serum lipid profile, intraperitoneal glucose tolerance test, and liver cytosolic antioxidant enzyme levels were measured. The HDF supplement significantly decreased serum glucose concentration, increased insulin level, and improved glucose homeostasis in diabetic control rats. The total cholesterol and triglyceride concentrations in the serum and liver were markedly reduced by HDF treatment in STZ-diabetic rats. Moreover, low density lipoprotein (LDL)-, VLDL-, and high density lipoprotein (HDL)-cholesterol levels were ameliorated in HDF supplemented diabetic rats. Decreased fecal excretions of cholesterol, triglyceride, and bile acid in diabetic rats were significantly increased by HDF consumption. HDF supplement reversed the effects of the oxidative stress system of liver in diabetic rats. Lipid peroxidation of diabetic rats, assessed by thiobarbituric acid reactive substance (TBARS), as well as superoxide dismutase (SOD) activities were significantly increased, and glutathione contents were decreased in diabetic rats. HDF supplement reverted these parameters to near normal value. Our data suggest that HDF supplement could be used to improve the glucose and lipid metabolism as well as to reduce the imbalance between the generation of reactive oxygen species (ROS) and the scavenging enzyme activity in preventing diabetic complications. PMID- 19122301 TI - Oral administration of synthetic retinoid Am80 inhibits the development of type 1 diabetes in non-obese diabetic (NOD) mice. AB - Synthetic retinoid Am80 is a potent modulator of the immune system. Am80 is effective in various experimentally induced autoimmune disorders. The purpose of this study is to confirm its effect on non-obese diabetic (NOD) mice, which spontaneously develop autoimmune type 1 diabetes. Am80 was orally administered in feed to 6 NOD mice per group at a dose of 0 (control), 0.1 (low) or 1 (high) mg/kg/d for 19 weeks. During the experiment period, the high urine glucose levels were observed in 33% mice of the control and low Am80 groups, whereas any mouse in the high Am80 group did not show abnormal urine glucose level. Histological examination showed that the average score of insulitis severity in the low Am80 group was similar to that in the control; however in the high Am80 group, the score was significantly reduced compared to that in the control group. Similarly, the severity of lymphocyte infiltration in the submandibular glands showed a tendency to decrease in the high Am80 group, but not in the low Am80 group, compared to the control. These data strongly suggest that the development of type 1 diabetes in NOD mice can be inhibited by oral administration of Am80. PMID- 19122302 TI - Thrombus aspiration complicated by systemic embolization in patients with acute myocardial infarction. AB - Percutaneous coronary intervention (PCI) is the preferred treatment modality for patients with acute coronary syndrome, but the "no-reflow" phenomenon, primarily caused by distal embolism, has hampered the effectiveness of PCI as regards reperfusion of the myocardium. Thrombus aspiration is sometimes used to reduce the incidence of distal embolism, but potentially the procedure may be complicated by upstream thrombus migration and systemic embolism. Two cases of systemic embolism during thrombus aspiration are presented. One patient had embolism of the cerebral artery and the other embolism of the left radial artery. It is suggested that a large-lumen guiding catheter with a deep-seated position and gentle injection of contrast medium should be used for thrombus aspiration. PMID- 19122303 TI - Cardiac amyloidosis presumptively diagnosed as cardiac syndrome X. AB - A 75-year-old man with cardiac amyloidosis was presumptively diagnosed as having cardiac syndrome X. Early clinical presentation was repeated episodes of angina pectoris without angiographically significant stenosis of the coronary arteries, so the diagnosis of cardiac syndrome X was initially given. However, his anginal symptoms worsened despite appropriate medication. He developed congestive heart failure and died 8 years after his first anginal symptoms. Autopsy revealed amyloid deposits in the intramyocardial coronary arteries with luminal obstruction, with little in the epicardial coronary arteries or myocardium. Therefore, the definitive diagnosis was cardiac amyloidosis. PMID- 19122304 TI - Fulminant eosinophilic myocarditis associated with visceral larva migrans caused by Toxocara canis infection. AB - A 19-year-old man was transferred to hospital because of myocarditis with cardiogenic shock. Echocardiography showed a left ventricular ejection fraction of 23.8% and an intermediate amount of pericardial effusion. The patient immediately received an intra-aortic balloon pump and percutaneous cardiopulmonary support. Right ventricular endomyocardial biopsy was performed in the acute phase and showed extensive eosinophilic inflammatory cell infiltration, severe interstitial edema and moderate myocardial necrosis. High-dose corticosteroids were administered. Because the patient's antibody titer against Toxocara canis was high and his symptoms had appeared after eating raw deer meat, the diagnosis was fulminant eosinophilic myocarditis caused by a hypersensitivity reaction to visceral larval migrans. After starting high-dose corticosteroids, the ejection fraction dramatically improved, the eosinophilia decreased and the patient made a full recovery. PMID- 19122305 TI - Two patients with bepridil-induced interstitial pneumonia. AB - Two patients developed bepridil-induced interstitial pneumonia during treatment of arrhythmia. The first patient was a 69-year-old man who received bepridil to maintain sinus rhythm in atrial fibrillation and who developed dyspnea on the 20th day after administration. The second patient was a 72-year-old man who received bepridil for paroxysmal atrial fibrillation and who developed dyspnea on the 60th day after administration. They were diagnosed with interstitial pneumonia on the basis of physical and imaging findings. The first patient was discharged after steroid pulse therapy, and the second patient after improvement of physical and imaging findings when bepridil was discontinued. Although a limited number of cases of bepridil-induced interstitial pneumonia have been reported, the disorder should be kept in mind as an important adverse reaction when breathlessness or dyspnea develops during administration of the drug. PMID- 19122306 TI - Carvedilol reduces the severity of central sleep apnea in chronic heart failure. AB - Background Carvedilol may reduce the severity of central sleep apnea (CSA) in patients with chronic heart failure (CHF). Methods and Results This study prospectively examined the effect of carvedilol on the severity of CSA in patients with CHF. Polysomnographic findings, left ventricular (LV) function, and plasma brain natriuretic peptide (BNP) level were evaluated before and 6 months after induction of carvedilol in 16 patients with CHF (New York Heart Association functional class II or III and LV ejection fraction <50%) who had CSA (central apnea index [CAI] >5 with dominant central apneic events). All patients tolerated carvedilol. The 6-month treatment with carvedilol increased the LV ejection fraction (32+/-7.4% to 45+/-9.8%, P<0.001) and decreased the BNP level (159 [69 458] pg/ml to 38 [16-193] pg/ml, P=0.017). The polysomnographic findings showed that the treatment decreased the apnea-hypopnea index (34+/-13 to 14+/-13, P=0.003) and CAI (13+/-11 to 1.9+/-4.3, P<0.001), whereas it increased the obstructive apnea index (1.1+/-1.5 to 3.1+/-3.4, P=0.04). Conclusion This preliminary study shows that treatment with carvedilol reduces the severity of CSA in patients with CHF, but that episodes of obstructive sleep apnea sometimes increase after the treatment. PMID- 19122307 TI - Detection of relevant coronary artery disease using dual-source computed tomography in a high probability patient series: comparison with invasive angiography. AB - BACKGROUND: Computed tomography (CT) enables detection of coronary artery stenoses, but its use is limited by deficient evaluation at elevated heart rates. The accuracy of dual-source CT (DSCT) for the detection of coronary artery disease (CAD) was assessed in 76 patients at high probability of CAD without heart rate control and compared with quantitative coronary angiography (QCA). Methods and Results The 76 patients (47 males, mean age 65.5+/-10 years) underwent DSCT without preceding heart rate control. Data sets were evaluated by 2 observers in consensus with respect to stenoses >50% decreased diameter. QCA served as the standard of reference. Mean heart rate during scanning was 68+/-9 beats per min, and the average Agatston score was 337+/-560. Of 1,160 coronary artery segments, all but 3 were visualized artefact-free; 58 coronary stenoses were correctly detected by CT angiography. In the segment-based analysis, sensitivity was 98.3%, specificity 99.2% and accuracy 99%; patient based analysis revealed a sensitivity of 100%, specificity of 83.3% and overall accuracy of 92.1%. Conclusions Even at elevated heart rates, DSCT can reliably detect coronary artery stenoses and the results correlate well with those for invasive coronary angiography. PMID- 19122308 TI - Effectiveness of nocturnal home oxygen therapy to improve exercise capacity, cardiac function and cardiac sympathetic nerve activity in patients with chronic heart failure and central sleep apnea. AB - BACKGROUND: Central sleep apnea, often found in patients with chronic heart failure (CHF), has a high risk of poor prognosis. METHODS AND RESULTS: This study involved 20 patients with CHF (left ventricular ejection fraction (LVEF) <45%, M/F =19/1, age 65+/-10 years) and an apnea-hypopnea index (AHI) >5 times/h who were divided into 2 groups: 10 patients treated with nocturnal home oxygen therapy (HOT) and 10 patients without HOT (non-HOT). All patients had dilated cardiomyopathy and underwent overnight polysomnography, cardiopulmonary exercise testing, and nuclear cardiac examinations to evaluate AHI, exercise capacity according to the specific activity scale and oxygen uptake at anaerobic threshold and peak exercise (peak VO(2)). Cardiac function according to (99m)Tc-MIBI QGS, and the total defect score (TDS), H/M ratio and the washout rate (WR) on (123)I metaiodobenzylguanidine (MIBG) imaging were calculated for all patients. As compared with the non-HOT group, the HOT group demonstrated a greater reduction in AHI (26.1+/-9.1 to 5.1+/-3.4), (123)I-MIBG TDS (31+/-8 to 25+/-9), and (123)I MIBG WR (48+/-8% to 41+/-5%) and a greater increase in the specific activity scale (4.0+/-0.9 to 5.8+/-1.2 Mets), peak VO(2) (16.0+/-3.8 to 18.3+/-4.7 ml . min(-1) . kg(-1)), and LVEF (27+/-9% to 37+/-10%). CONCLUSIONS: HOT improves exercise capacity, cardiac function, and cardiac sympathetic nerve activity in patients with CHF and central sleep apnea. PMID- 19122309 TI - Three new saponins from the fresh rhizomes of Polygonatum kingianum. AB - Further studies on the fresh rhizomes of Polygonatum kingianum led to the isolation of one new spirostanol saponin (25R)-kingianoside G (1), and two pairs mixture of 25R and 25S stereoisomeric spirostanol saponins (25R, S)-pratioside D1 (2a, 2b) and (25R, S)-kingianoside A (3a, 3b), among them 2b and 3b were new spirostanol saponins, together with another two known compounds, disporopsin (4) and daucosterol (5). The structures of the new saponins were determined by detailed analysis of their 1D and 2D NMR spectra, and chemical evidences. PMID- 19122310 TI - Heptacoordinate tin(IV) compounds derived from pyridine Schiff bases: synthesis, characterization, in vitro cytotoxicity, anti-inflammatory and antioxidant activity. AB - Tin(IV) complexes 2a--q derived from pyridine Schiff bases were prepared and characterized. Four complexes of this series were evaluated in vitro against different carcinogenic cell lines; besides their anti-inflammatory and antioxidant properties were also tested. Combination of mass spectrometry, multinuclear NMR and X-ray diffraction techniques evidenced the formation of heptacoordinated monomeric species. The X-ray diffraction analysis of 2a, 2b, 2i, 2j and 2n led to establish the heptacoordination around the tin atom in solid state and also revealed that the ligand occupies the equatorial positions of the distorted pentagonal bipyramidal geometry and the two alkyl or aryl groups the axial positions. The in-vitro study for complexes 2a--d against six tumor cell lines showed varied antiproliferative activity, the IC50 for all tested complexes was lower than that of the cis-platin. Compounds 2a--d also exhibited anti inflammatory activity where complex 2c resulted to be more active (IC50=0.11 microM) than the indomethacin IC50=0.27 microM which was used as reference. The antioxidant activity in rat brain homogenate on inhibition of thiobarbituric acid reactive substances (TBARS) indicated that 2c (IC50=1.77 microM) is more active than the quercetine (4.11 microM) and alpha-tocopherol (IC50=569.09 microM). PMID- 19122311 TI - Synthesis and evaluation of benzophenone oximes derivatized with sydnone as inhibitors of secretory phospholipase A2 with anti-inflammatory activity. AB - A series of benzophenone oximes appended with sydnone (3a--h) bearing different substituents on aroyl moiety were synthesized to evaluate in vivo and in vitro for their inhibitory activity against purified phospholipase A2 (PLA2) enzymes from snake venom and human inflammatory pleural and ascites fluid. In vivo and in vitro inhibition studies were carried out against PLA2 with respect to the modification of the pharmacophore (substituent) to analyze the specificity for PLA2. The substituent at the aroyl ring was responsible for enhancing the inhibition towards PLA2 enzymes. Most of the newly synthesized compounds inhibit the purified PLA2 enzyme, and the inhibition was more in hydrophobic and aromatic substituents and less when no such substituents were present. The inhibitory effect of the compounds appeared to be due to the direct interaction of compounds with the enzyme. Inhibition is substrate dependent, and the inhibition competes with the substrate for the same binding site of the enzyme. The most active interacting compound 3h from in vitro inhibition of PLA2 activity showed similar potency in the in vivo neutralization of PLA2 induced mouse paw edema and hemolytic activity. Thus, the in vitro inhibition correlated well with the in vivo inhibition and hence the reported derivatives are therapeutically important anti-inflammatory drugs. PMID- 19122312 TI - Discovery of R-142086 as a factor Xa (FXa) inhibitor: syntheses and structure activity relationships of cinnamyl derivatives. AB - To develop a novel and effective anticoagulant with potent and selective factor Xa (FXa) inhibitory activity, a new series of cinnamyl derivatives with enhanced lipophilicity and prodrug forms were synthesized and their biological activities were evaluated. As a result, we found that cinnamyl derivative (N-[4-[1 (acetimidoyl)piperidin-4-yloxy]-3-carbamoylphenyl]-N-[(Z)-3-(3-amidinophenyl)-2 fluoro-2-propenyl]sulfamoyl)acetic acid dihydrochloride (26d, R-142086) with a fluorine atom on the double bond exhibited potent anticoagulant activity and no mutagenic potential. Moreover, orally administered R-142086 exhibited potent anti FXa activity and anticoagulant activity in dogs. PMID- 19122313 TI - Design and synthesis of piperazinylpyridine derivatives as novel 5-HT1A agonists/5-HT3 antagonists for the treatment of irritable bowel syndrome (IBS). AB - We have prepared a series of piperazinylpyridine derivatives for the treatment of irritable bowel syndrome (IBS). These compounds, which were designed by pharmacophore analysis, bind to both serotonin subtype 1A (5-HT1A) and subtype 3 (5-HT3) receptors. The nitrogen atom of the isoquinoline, a methoxy group and piperazine were essential to the pharmacophore for binding to these receptors. We also synthesized furo- and thienopyridine derivatives according to structure activity relationship analyses. Compound 17c (TZB-20810) had high affinities to these receptors and exhibited 5-HT1A agonistic activity and 5-HT3 antagonistic activity concurrently, and is a promising drug for further development in the treatment of IBS. PMID- 19122314 TI - Freeze-drying of proteins in glass solids formed by basic amino acids and dicarboxylic acids. AB - The purpose of this study was to produce and characterize glass-state amorphous solids containing amino acids and organic acids that protect co-lyophilized proteins. Thermal analysis of frozen solutions containing a basic amino acid (e.g., L-arginine, L-lysine, L-histidine) and a hydroxy di- or tricarboxylic acid (e.g., citric acid, L-tartaric acid, DL-malic acid) showed glass transition of maximally freeze-concentrated solute at temperatures (T'g) significantly higher than those of the individual solute solutions. Mixing of the amino acid with some dicarboxylic acids (e.g., oxalic acid) also suggested an upward shift of the transition temperature. Contrarily, combinations of the amino acid with monocarboxylic acids (e.g., acetic acid) had T'gs between those of the individual solute solutions. Co-lyophilization of the basic amino acids and citric acid or L tartaric acid resulted in amorphous solids that have glass transition temperatures (Tg) higher than the individual components. Mid- and near-infrared analysis indicated altered environment around the functional groups of the consisting molecules. Some of the glass-state excipient combinations protected an enzyme (lactate dehydrogenase, LDH) from inactivation during freeze-drying. The glass-state excipient combinations formed by hydrogen-bonding and electrostatic interaction network would be potent alternative to stabilize therapeutic proteins in freeze-dried formulations. PMID- 19122315 TI - An alternative convenient synthesis of piperidazine-3-carboxylic acid derivatives. AB - The short-step synthesis of the unsubstituted, 5-hydroxy- and 5 chloropiperidazine-3-carboxylic acids using an aza Diels-Alder reaction between the 1,3-diene and azodicarboxylate was described. This synthetic methodology could be used for the preparation of the optically active piperazic acid in a 35% overall yield. PMID- 19122316 TI - Design and evaluation of pH-independent pulsatile release pellets containing isosorbide-5-mononitrate. AB - A three-layered, pH-independent pulsatile release pellets system containing isosorbide-5-mononitrate (ISMN) was studied. The process of the heart disease such as angina has a close relationship to the chronobiology, which gives rise to the need of a pulsatile drug deliver system for the anti-anginal drug. In this study, pellets containing ISMN were firstly prepared as the core, and then layered with a swelling layer followed by an water-insoluble control layer. The core pellets were formulated with microcrystalline cellulose (MCC) and lactose, and were prepared by extrusion-spheronization. The preparation was optimized by Box-Behnken experimental design, when taking the MCC/lactose ratio as swell as the operating conditions of extrusion-spheronization as variables. The experimental results demonstrated the relationships between formulation, operation and properties of the product, and meanwhile provided optimized values for the parameters. The core pellets were coated by a fluidized bed coater, and pellets with various coating types and coating levels were studied by in vitro dissolution tests. The effects of both swelling layer and control layer on the lag time and the drug release time were studied, in order to predetermine the lag time and release time. The pellets were also evaluated in vivo by studying the pharmacokinetics after oral administration in beagle dogs. The pellets achieved a lag time of 4.1 h in vivo, which had a good consistency with the in vitro results, and the relative bioavailability was nearly 100% comparing to the normal tablets. PMID- 19122317 TI - Feasibility of 19F-NMR for assessing the molecular mobility of flufenamic acid in solid dispersions. AB - The purpose of the present study was to clarify the feasibility of 19F-NMR for assessing the molecular mobility of flufenamic acid (FLF) in solid dispersions. Amorphous solid dispersions of FLF containing poly(vinylpyrrolidone) (PVP) or hydroxypropylmethylcellulose (HPMC) were prepared by melting and rapid cooling. Spin-lattice relaxation times (T1 and T(1rho)) of FLF fluorine atoms in the solid dispersions were determined at various temperatures (-20 to 150 degrees C). Correlation time (tauc), which is a measure of rotational molecular mobility, was calculated from the observed T1 or T1rho value and that of the T1 or T1rho minimum, assuming that the relaxation mechanism of spin-lattice relaxation of FLF fluorine atoms does not change with temperature. The tauc value for solid dispersions containing 20% PVP was 2-3 times longer than that for solid dispersions containing 20% HPMC at 50 degrees C, indicating that the molecular mobility of FLF in solid dispersions containing 20% PVP was lower than that in solid dispersions containing 20% HPMC. The amount of amorphous FLF remaining in the solid dispersions stored at 60 degrees C was successfully estimated by analyzing the solid echo signals of FLF fluorine atoms, and it was possible to follow the overall crystallization of amorphous FLF in the solid dispersions. The solid dispersion containing 20% PVP was more stable than that containing 20% HPMC. The difference in stability between solid dispersions containing PVP and HPMC is considered due to the difference in molecular mobility as determined by tauc. The molecular mobility determined by 19F-NMR seems to be a useful measure for assessing the stability of drugs containing fluorine atoms in amorphous solid dispersions. PMID- 19122318 TI - New neolignan component from Camellia amplexicaulis and effects on osteoblast differentiation. AB - A new neolignan named "camellioside A" (1) has been isolated from the leaves and branches of Camellia amplexicaulis, along with seven known glycosides (2-8). Their structures were determined by a variety of spectroscopic analyses. Among them, compounds 2, 3, 4, and 6 significantly (p<0.05) increased the alkaline phosphatase activity and the mineralization of the nodules of the MC3T3-E1 osteoblastic cells compared to those of the control. PMID- 19122319 TI - Synthesis, characterization and DNA-binding properties of the Cu(II) complex with 7-methoxychromone-3-carbaldehyde-benzoylhydrazone. AB - A new chromone derivative (7-methoxychromone-3-carbaldehyde-benzoylhydrazone) ligand (HL) and its novel Cu(II) complex have been synthesized and characterized on the basis of elemental analyses, molar conductivities, proton nuclear magnetic resonance, infrared spectra and mass spectra analyses. The general formula of the Cu(II) complex is [CuL(HB2BO)]Cl x 2H2O. In addition, the interaction of the Cu(II) complex and its free ligand with calf-thymus DNA was investigated by electronic absorption spectroscopy, fluorescence spectroscopy and viscosity measurement. Results suggest that both the two compounds can bind with calf thymus DNA via an intercalation mechanism. Furthermore, the Cu(II) complex can bind to DNA more strongly than the free ligand due to the chelating effect of copper(II) ion to the free ligand. PMID- 19122320 TI - Syntheses and doxorubicin-inclusion abilities of beta-cyclodextrin derivatives with a hydroquinone alpha-glycoside residue attached at the primary side. AB - This paper describes syntheses and doxorubicin-inclusion abilities of beta cyclodextrin (CyD) derivatives with a hydroquinone alpha-glycoside residue attached at the primary side. The hydroquinone glycoside having an alpha-D glucosidic or 2-acetamido-2-deoxy-alpha-D-glucosidic linkage became a useful component for providing an alpha-D-glucose- or 2-acetamido-2-deoxy-alpha-D glucose-beta-CyD conjugate. The surface plasmon resonance analyses of these beta CyD derivatives for the anticancer agent, doxorubicin, indicated that they had excellent inclusion associations on the order of 10(5 m)-1 for the immobilized doxorubicin. PMID- 19122321 TI - Determination of sweroside in rat plasma and bile for oral bioavailability and hepatobiliary excretion. AB - Sweroside is an active ingredient of iridoid glycoside isolated from the flower buds of Lonicera japonica THUNB. A quantitative HPLC-UV method was developed for monitoring sweroside in rat plasma, urine, feces and bile. The method was successfully applied for a basic pharmacokinetic study. The obtained data of pharmacokinetics were applied to evaluate the oral bioavailabilities of sweroside and the active ingredients of purified herbal extracts (IGEs-1). The absolute bioavailability was estimated to F(sweroside) 0.31% and F(IGEs-1) 0.67%. The majority of sweroside excreted to feces revealed one reason of the low oral bioavailability. The values of F(IGEs-1) much higher than that of F(sweroside) reveals that ingredients in IGEs-1 such as loganin, secoxyloganin and some phenolic acids may promote the absorption of sweroside. The study of hepatobiliary excretion was achieved by an in vivo microdialysis sampling method after intravenous administration of sweroside. The percentage of accumulation of free form sweroside in bile duct was 31.2+/-7.2% of the total dosage. It may be one reason why sweroside possesses strong hepatoprotective effect. PMID- 19122322 TI - Anti human immunodeficiency virus-1 (HIV-1) agents 1. Discovery of benzyl phenyl ethers as new HIV-1 inhibitors in vitro. AB - Ten single benzyl phenyl ethers were synthesized and evaluated as human immunodeficiency virus-1 (HIV-1) inhibitors in vitro for the first time. Among these compounds, especially 4-nitrobenzyl phenyl ether (3h) exhibited the highest anti-HIV-1 activity with EC50 (concentration of drug that reduces syncytia formation by 50%) value of 5.96 microg/ml and therapeutic index value of 18.32. The preliminary structure-activity relationships of these benzyl phenyl ethers were also described. PMID- 19122323 TI - Alkaloids from the South China Sea Black Coral Antipathes dichotoma. AB - A new carbazole alkaloid, antipathine A (1), together with three known zoanthoxanthin alkaloids (2--4) was isolated from the EtOH/CH2Cl2 extracts of the South China Sea black coral Antipathes dichotoma. The structure of 1 was determined on the bases of extensive spectroscopic analysis, including 1D and 2D NMR data. Compounds 1 and 2 showed moderate cytotoxicity against human stomach carcinoma SGC-7901 cell line with IC50 of 67.38 and 86.40 microg/ml, respectively, and 1 and 2 also showed weak cytotoxicity toward human liver carcinoma Hep_G2 cell line. PMID- 19122324 TI - Acetylation of the amino group on guanosine induced by nitric oxide in acetonitrile under aerobic conditions. AB - When nitric oxide was bubbled into acetonitrile under aerobic conditions, the solution showed a cobalt-blue color. Addition of guanosine into the solution generated N2-acetylguanosine as a major product. The result of the reaction using 15N labeled acetonitrile indicated that the nitrogen atom of the acetylated exocyclic amino group on N2-acetylguanosine originated from acetonitrile. We discuss the reaction mechanism for the acylation. PMID- 19122325 TI - Ginsenosides from heat processed ginseng. AB - A new dammarane-glycoside, ginsenoside Rz1 (1), was isolated from heat-treated Panax ginseng C. A. MEYER (Araliaceae) with ginsenosides Rk1 and Rg5. The structure of 1 was established to be (Z)-12beta-hydroxydammara-20(22),24-dien 3beta-yl O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucospyranoside by spectroscopic means. HPLC analyses revealed that the ginsenosides Rz1, Rk1, and Rg5 were present in the ratios of 1 : 2 : 6, respectively. PMID- 19122326 TI - Synthesis of a novel water-soluble cleft-type cyclophane as an N-methyl-D aspartate receptor antagonist. AB - Novel water-soluble N-methyl-D-aspartate (NMDA) receptor antagonists, 4,4'-bis([2 [N-(1,4,8,11-tetraazacyclotetradecan-1-yl)acetyl]-N phenethyl]aminoethoxy)diphenylmethane octahydrochloride (1, ACPCm) and 4,4' bis([2-[N-(1,4,7,10-tetraazacyclododecan-1-yl)acetyl]-N phenethyl]aminoethoxy)diphenylmethane octahydrochloride (2, ACPCn), were synthesized and the effect of these cleft-type cyclophanes on NMDA receptors was then studied using voltage-clamp recordings of recombinant NMDA receptors expressed in Xenopus oocytes. ACPCm (1) and ACPCn (2) inhibited macroscopic currents in the NR1/NR2A, NR1/NR2B, NR1/NR2C and NR1/NR2D receptor subtypes in oocytes voltage-clamped at -70 mV. The IC50 values of ACPCm (1) and ACPCn (2) for NR1/NR2A and NR1/NR2B receptors were 1.06 microM and, 0.92 microM and 1.47 microM and, 1.49 microM, respectively. The inhibition by these compounds was voltage dependent, that is, the degree of inhibition was in the order of negative holding potentials, -100 mV>-70 mV>-20 mV. These findings indicate that the cleft-type cyclophanes, ACPCm (1) and ACPCn (2) directly act on the channel pore of the NMDA receptors. PMID- 19122327 TI - Cordyceamides A and B from the Culture Liquid of Cordyceps sinensis (BERK.) SACC. AB - Two new aurantiamides named as cordyceamides A and B were isolated from the culture liquid of Cordyceps sinensis (BERK.) SACC., along with one known compound, aurantiamide acetate. Their structures were elucidated as N-benzoyl-L tyrosinyl-L-phenylalaninol acetate and N-benzoyl-L-tyrosinyl-L-p hydroxyphenylalaninol acetate by 1D, 2D-NMR techniques and comparison with literatures. PMID- 19122328 TI - New triterpenoid saponins from Glochidion eriocarpum and their cytotoxic activity. AB - Combined chromatographic methods led to the isolation of two new triterpenoid saponins, glochieriosides A and B (1, 2), from the aerial parts of Glochidion eriocarpum, along with three known triterpenes, glochidone (3), lup-20(29)-en 3beta,23-diol (4), and lup-20(29)-en-1beta,3beta-diol (5). The structures of the new saponins were determined to be 22beta-benzoyloxy-3beta,16beta,28 trihydroxyolean-12-ene 3-O-[beta-D-glucopyranosyl-(1-->3)-alpha-L arabinopyranoside] (1) and 22beta-benzoyloxy-3beta,16beta,28-trihydroxyolean-12 ene 3-O-[beta-D-glucopyranosyl-(1-->3)-beta-D-xylopyranoside] (2). The structural elucidation was accomplished by using a combination of the 1D-NMR (1H-, 13C-NMR, distortionless enhancement by polarization transfer (DEPT) 90 degrees , and DEPT 135 degrees ), 2D-NMR (1H-1H correlation spectroscopy, heteronuclear multiple quantum correlation, heteronuclear multiple bond correlation, and rotating frame Overhouser effect spectroscopy), ESI-MS, and HR-FAB-MS experiments. Glochieriosides A and B exhibited significant cytotoxic activity against HL-60, HT-29, MCF-7 and SK-OV-3 human cancer cell lines with the IC50 values of 5.5, 6.8, 29.1, and 22.7 microM for glochierioside A, respectively, and 6.6, 18.6, 36.1, and 16.0 microM for glochierioside B. Glochidone was less active with IC50 values greater than 100 microM while lup-20(29)-en-1beta,3beta-diol was moderately active with IC50 values of 43.3, 67.0, 66.1, and 48.0 microM, respectively. PMID- 19122329 TI - New cerebrosides from a marine sponge Haliclona (Reniera) sp. AB - A chemical investigation of the MeOH extract of a marine sponge Haliclona (Reniera) sp., collected off the coast of Ulleung Island, Korea, led to the isolation of thirteen new cerebrosides (1--3, 5--14), along with a known analogue (4). Their structures were elucidated on the basis of 1D and 2D NMR spectroscopy, MS spectrometry, and chemical method. The major new features of these glucocerebrosides are C15 and C19 acyl chains, long (C24-C28) acyl chains, or the S-configuration of the acyl chains. It is noteworthy that both R- and S configurations of the acyl chains were observed in the same specimen. PMID- 19122330 TI - Iodine-catalyzed etherification of morroniside. AB - In this study, we describe a highly selective etherification procedure of unprotected morroniside catalyzed by molecular iodine in acetone. The etherification reaction furnished 7-O-alkyl ether derivatives in reasonable yields within few hours under neutral conditions. Studies of the obtained products on cytotoxicity activity in colon 26-L5 cell line were examined. Among the tested compounds, 7-O-dodecylmorroniside showed moderate cytotoxic activity, having IC50 values equal to 20.9 microM. PMID- 19122331 TI - pXBP1(U), a negative regulator of the unfolded protein response activator pXBP1(S), targets ATF6 but not ATF4 in proteasome-mediated degradation. AB - Cells from yeast to humans activate unconventional mRNA splicing when unfolded proteins accumulate in the endoplasmic reticulum (ER) under ER stress conditions. The substrate of this splicing in mammalian cells is XBP1 mRNA, which encodes the unfolded protein response (UPR)-specific transcription factor XBP1. The C terminal region of XBP1 is switched as a result of the splicing. Thus, unspliced and spliced mRNAs produce pXBP1(U) of 261 aa and pXBP1(S) of 376 aa, respectively, with the N-terminal region containing the DNA-binding domain shared. As the pXBP1(S)-specific C-terminal region functions as an activation domain, pXBP1(S) can activate transcription efficiently. We recently found that pXBP1(U) shuttles between the nucleus and cytoplasm, owing to the presence of a nuclear exclusion signal in the pXBP1(U)-specific C-terminal region, in marked contrast to the exclusively nuclear localization of pXBP1(S). pXBP1(U) can associate with pXBP1(S), and pXBP1(U)-pXBP1(S) complex is rapidly degraded by the proteasome. Two other transcription factors are activated in response to ER stress, namely ATF6 and ATF4. ATF6 is a UPR-specific transcription factor, whereas ATF4 is activated by not only ER stress but also various other stimuli. In this study, we show that pXBP1(U) targets the active form of ATF6 but not ATF4 for destruction by the proteasome via direct association. This enhanced degradation is mediated by the degradation domain located at the pXBP1(U) specific C-terminal end. We conclude that pXBP1(U) functions as a negative regulator of the UPR-specific transcription factors ATF6 and pXBP1(S). PMID- 19122333 TI - ABCC2/Abcc2 transport property in different species and its modulation by heterogeneous factors. AB - ABCC2/Abcc2 is a member of the ABC transporter family expressed mainly in the liver bile canalicular membrane and involved in the excretion of various kinds of organic anions from hepatocytes into bile. During the drug development process, species differences in the pharmaco- and toxicokinetics of candidate drugs are a major problem. It is possible that ABCC2/Abcc2 transport activity as well as inhibitor sensitivity could lead to a number of phenomena (e.g. a difference in the biliary excretion clearance, a delay in the elimination half-life from the circulating blood and toxic side effects on ABCC2 -mediated drug-drug interactions, such as drug-induced hyperbilirubinemia). From this point of view, it is useful to be able to predict during preclinical development if certain compounds of interest are substrates and/or modulators of ABCC2. Although an in vivo animal model or an in vitro model expressing ABCC2 are useful assay systems, these have some limitations as far as predicting the transport profile of compounds in vivo is concerned. I will present an overview of the species differences in the tissue distribution, function, and also characteristic transport properties of ABCC2/Abcc2 mainly in an in vitro experimental model. PMID- 19122334 TI - Influence of SLCO1B3 gene polymorphism on the pharmacokinetics of digoxin in terminal renal failure. AB - Digoxin (DX) is mainly excreted unchanged in the urine. In patients undergoing hemodialysis (HD patients), the relative contribution of hepatic elimination is increased. DX is a substrate of OATP1B3 (SLCO1B3), expressed on the sinusoidal membranes of hepatocytes in humans. Therefore, we investigated the relationship between SLCO1B3 gene polymorphisms and the value of trough concentration-to-dose ratio (C/D ratio) of DX in HD patients. We investigated two deletion polymorphisms in complete linkage disequilibrium (-28 to -11 and -7 to -4) and two SNPs in complete linkage disequilibrium (T334G and G699A). Blood was sampled 62-72 hours after the oral administration of DX. The C/D ratio of DX was lower in patients with the deletion allele than in those homozygous for the insertion allele, and was lower in patients with the 334T/669G allele than in those homozygous for the 334G/699A allele, although the differences were not statistically significant. The C/D ratio of DX was significantly higher in patients with homozygous for the insert-variant allele (median: 121.8 (ng/mL)/(mg/week/kg), range: 92.5-259.4 (ng/mL)/(mg/week/kg) than in others (median: 93.4 (ng/mL)/(mg/week/kg), range: 66.5-154.3 (ng/mL)/(mg/week/kg)). In conclusion, the insert-variant allele of the OATP1B3 gene may increase the C/D ratio of DX in HD patients. PMID- 19122335 TI - Comparison of human cytochrome P450 inhibition by the thienopyridines prasugrel, clopidogrel, and ticlopidine. AB - Differences in the inhibition of cytochrome P450 activities among thienopyridine antiplatelet agents, ticlopidine, clopidogrel, prasugrel, and the metabolites, 2 oxo-clopidogrel, clopidogrel acid metabolite, deacetylated metabolite of prasugrel (R-95913) and the pharmacologically active metabolites of clopidogrel and prasugrel, were examined using recombinant cytochromes P450 and fluorescent probe substrates. Ticlopidine and clopidogrel inhibited CYP2B6 with IC(50) values of 0.0517+/-0.0323 microM and 0.0182+/-0.0069 microM, respectively, and inhibited CYP2C19 with IC(50) values of 0.203+/-0.124 microM and 0.524+/-0.160 microM, respectively. Ticlopidine also inhibited CYP2D6 (IC(50) of 0.354+/-0.158 microM). In contrast, 2-oxo-clopidogrel, prasugrel and R-95913 were much weaker inhibitors of CYP2B6, CYP2C19 and CYP2D6. The inhibitory effects of all the compounds tested were much weaker on the isoforms other than those indicated above. The active metabolites of clopidogrel and prasugrel and clopidogrel acid metabolite also did not affect the activities of the P450s examined. PMID- 19122336 TI - Time-dependent induction of rat hepatic CYP1A1 and CYP1A2 expression after single dose administration of the anti-angiogenic agent TSU-68. AB - The anti-angiogenic agent TSU-68 is known to rapidly induce cytochrome P450 activity responsible for its own hydroxylation in rats. In this study, we identified CYP1A1 and CYP1A2 as the TSU-68-induced P450 and temporally characterized the rapid induction of these isoforms. Protein and mRNA levels of CYP1A1 and CYP1A2 along with CYP1A activities were examined in rat liver after a single oral administration of 500 mg/kg TSU-68. CYP1A-mediated ethoxyresorufin O deethylation and TSU-68 hydroxylation activities reached the maximum at 12 hr. The activities were maintained up to 24 hr and then slowly decreased down to control levels. Protein levels of both CYP1A1 and CYP1A2 were also rapidly induced with temporal profiles similar to the profile of CYP1A activity. In contrast, unlike CYP1A2 mRNA levels, which peaked at 12 hr and almost returned to control levels by 48 hr, CYP1A1 mRNA levels peaked as early as 3 hr and returned to control levels by 24 hr. Thus, CYP1A1 showed more rapid elevation and turnover of its mRNA than CYP1A2. In conclusion, TSU-68 administered to rats rapidly induced mRNA and protein of CYP1A1 and CYP1A2 as well as CYP1A activity. Furthermore, the data showed a difference in the time-dependent induction between CYP1A1 and CYP1A2 mRNAs. PMID- 19122337 TI - Effect of clarithromycin on the pharmacokinetics of pranlukast in healthy volunteers. AB - Pranlukast is a cysteinyl leukotriene receptor antagonist that has been used to treat bronchial asthma and allergic rhinitis. In vitro data suggest that pranlukast is a substrate of CYP3A4. Thus, the effect of clarithromycin, a potent CYP3A4 inhibitor, on the pharmacokinetics of pranlukast was examined in an open label, randomized, two-way crossover study in 16 healthy male volunteers. In treatment A, volunteers received a single, 225 mg dose of pranlukast. In treatment B, 200 mg of clarithromycin was administered twice daily for 7 days and a single, 225 mg dose of pranlukast was coadministered on day 7. Blood samples were collected up to 24 hours after treatment, and pranlukast concentrations in the plasma were measured. The geometric mean ratios [GMR] (90% confidence intervals [CIs]) for pranlukast AUC(0-infinity) and C(max) (with/without clarithromycin) were 1.06 (0.91, 1.24) and 1.17 (0.95, 1.45), respectively. In conclusion, clarithromycin and pranlukast could be coadministered without dose adjustment because clarithromycin minimally affected the pharmacokinetics of pranlukast. PMID- 19122338 TI - Population pharmacokinetics of tebipenem pivoxil (ME1211), a novel oral carbapenem antibiotic, in pediatric patients with otolaryngological infection or pneumonia. AB - Tebipenem pivoxil (TBPM-PI, ME1211) has been under development as the world's first oral carbapenem for treatment of otolaryngological/respiratory infections caused by drug-resistant S. pneumoniae in pediatric patients. In order to treat these infections effectively, it is important to design optimal dosing regimens based on the pharmacokinetics/pharmacodynamics (PK/PD) relationships, which can be characterized by clarifying the pharmacokinetics of tebipenem (TBPM) in the pediatric population. We therefore performed an population pharmacokinetic analysis using plasma TBPM concentrations obtained from pediatric patients with otolaryngological infection or bacterial pneumonia (0.5-16 years old; n=217, 395 points), after repeated oral administration of TBPM-PI at a dose of 4 or 6 mg/kg b.i.d. A one-compartment model with first-order absorption was adopted. In analysis, weight-normalized creatinine clearance (Ccr) and age were the most significant covariates that respectively explained inter-subject variability in weight-normalized apparent clearance (CL/F) and volume of distribution (Vd/F) of TBPM. The CL/F of TBPM increased with Ccr, and the Vd/F decreased with age. Based on the results of the present analysis, validity of the presently recommended dosage regimen of TBPM-PI in pediatric patients is discussed. PMID- 19122339 TI - Population pharmacokinetics of landiolol hydrochloride in healthy subjects. AB - Landiolol hydrochloride is a newly developed cardioselective, ultra short-acting beta(1)-adrenergic receptor blocking agent used for perioperative arrhythmia control. The objective of this study was to characterize the population pharmacokinetics of landiolol hydrochloride in healthy male subjects. A total of 420 blood concentration data points collected from 47 healthy male subjects were used for the population pharmacokinetic analysis. NONMEM was used for population pharmacokinetic analysis. In addition, the final pharmacokinetic model was evaluated using a bootstrap method and a leave-one-out cross validation method. The concentration time course of landiolol hydrochloride was best described by a two-compartment model with lag time. The final parameters were total body clearance (CL: 36.6 mL/min/kg), distribution volume of the central compartment (V1: 101 mL/kg), inter-compartmental clearance (16.1 mL/min/kg), distribution volume of the peripheral compartment (55.6 mL/kg), and lag time (0.82 min). The inter-individual variability in the CL and V1 were 21.8% and 46.3%, respectively. The residual variability was 22.1%. Model evaluation by the two different methods indicated that the final model was robust and parameter estimates were reasonable. The population pharmacokinetic model for landiolol hydrochloride in healthy subjects was developed and was shown to be appropriate by both bootstrap and leave-one-out cross validation methods. PMID- 19122340 TI - Population pharmacokinetic analysis of a novel muscarinic receptor antagonist, imidafenacin, in healthy volunteers and overactive bladder patients. AB - The objectives of this study were to develop a population pharmacokinetic model of imidafenacin and to explore the factors that affect the pharmacokinetics of imidafenecin. A total of 2406 plasma samples were collected from 90 healthy volunteers and 457 patients with overactive bladder. We determined the plasma concentrations of imidafenacin by liquid chromatography with tandem mass spectrometry; resultant data were analyzed by a population approach using NONMEM software. The imidafenacin plasma concentration time course was described using a two-compartment model with first-order absorption and lag time. The robustness of the population pharmacokinetic model was evaluated by bootstrap resampling. The results of the population pharmacokinetic analysis demonstrated that oral clearance was decreased with advancing age, increasing hepatic function parameters (AST and ALP), food intake, and itraconazole coadministration, while the first-order absorption rate constant was decreased with food intake. All parameter estimates from the final model fell within 20% of the bootstrapped mean. In conclusion, we developed a population pharmacokinetic model for imidafenacin that well-described plasma concentration profiles. We also identified the factors affecting imidafenacin pharmacokinetics. PMID- 19122341 TI - Inhibitory effect of zinc on the absorption of beta-lactam antibiotic ceftibuten via the peptide transporters in rats. AB - Zinc is an essential metal ion for the body, and is widely used for nutritional and clinical purposes. Previously, we showed that zinc inhibits the transport of glycylsarcosine via the intestinal peptide transporter PEPT1 in the human intestinal cell line Caco-2. In this study, we examined the effect of zinc on the activity of peptide transporters in rats using the oral beta-lactam antibiotic ceftibuten as a model drug. The plasma ceftibuten concentration after intraintestinal administration was decreased in the presence of zinc. The maximum plasma concentration (C(max)) was significantly decreased and the time required to reach C(max) (T(max)) was prolonged by zinc coadministration. The plasma ceftibuten concentration after iron coadministration or two hours after zinc administration was not affected. The in situ loop technique revealed 50% inhibition of ceftibuten absorption by zinc. In conclusion, zinc inhibits the transport activity of PEPT1 in vivo as well in vitro. PMID- 19122342 TI - Effect of intestinal and hepatic first-pass extraction on the pharmacokinetics of everolimus in rats. AB - The aim of this study was to quantitatively evaluate the effects of intestinal and hepatic extraction on the pharmacokinetics of everolimus in rats. Everolimus was administered intravenously, intraportally or intraintestinally in order to separately evaluate the intestinal and hepatic first-pass extraction. Cyclosporine or tacrolimus was administered into rat intestines, and after 10 min everolimus pharmacokinetics were evaluated. The blood concentrations of everolimus were measured by the high-performance liquid chromatography with tandem mass spectrometry. Total body clearance of everolimus was constant in the dosage from 0.2 to 1.0 mg/kg. The bioavailability after intraportal and intraintestinal administration were 48.0% and 21.2%, respectively. Concomitantly administered cyclosporine (5 mg/kg) , but not tacrolimus (1 mg/kg), significantly decreased the total body clearance of everolimus compared with the control, and also increased the bioavailabilty of everolimus after intraintestinal administration 1.75-fold. Cyclosporine significantly increased the area under the blood concentration-time curve of everolimus after the intraintestinal constant infusion 3-fold, and increased that after the intraportal constant infusion only 1.35-fold. In conclusion, the intestine as well as liver contributes to the first pass extraction for everolimus in rats. Intestinally administered cyclosporine inhibited the intestinal extraction of everolimus more than its hepatic extraction. PMID- 19122343 TI - Genetic variations of the SLCO1B1 gene in the Chinese, Malay and Indian populations of Singapore. AB - OATP1B1 is a liver-specific transporter that mediates the uptake of various endogenous and exogenous compounds including many clinically used drugs from blood into hepatocytes. This study aims to identify genetic variations of SLCO1B1 gene in three distinct ethnic groups of the Singaporean population (n=288). The coding region of the gene encoding the transporter protein was screened for genetic variations in the study population by denaturing high-performance liquid chromatography and DNA sequencing. Twenty-five genetic variations of SLCO1B1, including 10 novel ones, were found: 13 in the coding exons (9 nonsynonymous and 4 synonymous variations), 6 in the introns, and 6 in the 3' untranslated region. Four novel nonsynonymous variations: 633A>G (Ile211Met), 875C>T (Ala292Val), 1837T>C (Cys613Arg), and 1877T>A (Leu626Stop) were detected as heterozygotes. Among the novel nonsynonymous variations, 633A>G, 1837T>C, and 1877T>A were predicted to be functionally significant. These data would provide fundamental and useful information for pharmacogenetic studies on drugs that are substrates of OATP1B1 in Asians. PMID- 19122345 TI - An increase in serum retinol-binding protein 4 in the type 2 diabetic subjects with nephropathy. AB - The present study was undertaken to determine retinol-binding protein 4 (RBP4) levels in subjects with diabetic nephropathy. A total of 149 type 2 diabetic subjects and 19 control subjects were enrolled. Serum levels of RBP4 were measured by a method of ELISA. Serum RBP4 levels were significantly greater in the subjects with type 2 diabetes mellitus than the controls (70.5 +/- 35.3 vs. 40.1 +/- 13.0 microg/ml, mean +/- SD, p<0.01). Serum RBP4 levels were gradually increased according to the progression of diabetic nephropathy (p value in trend test: <0.001). Its elevation was significantly greater in the diabetic subjects with stages 1, 3B and 4 than the control subjects (Stage 1: 64.6 +/- 29.7, Stage 3B: 123.3 +/- 71.8, Stage 4: 91.4 +/- 33.8 vs. CONTROL: 40.1 +/- 13.0 microg/ml, p<0.01). Similar results were obtained in the subjects based on the amount of albuminuria (Normo-: 64.6 +/- 29.7, Micro-: 63.7 +/- 29.4, and Marcoalbuminuria: 90.3 +/- 44.6 microg/ml, p <0.001). Serum RBP4 levels had a positive correlation with serum creatinine levels(r = 0.377, p<0.001), and a negative correlation with 1/creatinine (r = -0.420, p<0.001). Also, there was a negative correlation between serum RBP4 and the estimated glomerular filtration rate(r = -0.436, p<0.001). Multiple regression analysis showed that estimated glomerular filtration rate was an independent determinant for increased serum RBP4 levels. There was no difference in serum RBP4 levels between the advanced nephropathy with and without macrovascular diseases. These results indicate an increase in serum RBP4 levels in the type 2 diabetic subjects, particularly complicated with advanced renal impairment. PMID- 19122346 TI - Identification of minimal promoter and genetic variants of Kruppel-like factor 11 gene and association analysis with type 2 diabetes in Japanese. AB - Genetic analysis of the KLF11 gene revealed two rare variants, A347S and T220M, segregating in families with early-onset type 2 diabetes, and one frequent polymorphic Q62R variant significantly associated with type 2 diabetes in Northern Europeans. Furthermore, it has been reported that over-expression of KLF11 has a deleterious effect on insulin promoter activity. Thus, an altered expression level of KLF11 may contribute to the occurrence of type 2 diabetes. To investigate the contribution of KLF11 to type 2 diabetes in Japanese, we surveyed the 5' flanking region of KLF11 by reporter assay and identified the minimal promoter region of the gene. The promoter region from -250 to +162 bp including five Sp1 binding sites showed basal promoter activity both in MIN6-m9 and HepG2 cells. We also examined the entire region of KLF11 to detect genetic variants. A total of 19 polymorphisms, six of which are novel, were identified, but none of them showed association with the occurrence of type 2 diabetes. Two of the identified polymorphisms, R29Q and S124F, are novel coding variants. Functional analyses of these variants were performed, and similarly reduced effects on transcriptional activities of insulin, catalase1, and the Smad7 gene were found. We conclude that variants of KLF11 are not a major factor in the occurrence of type 2 diabetes in Japanese. The promoter region of KLF11 identified in the present study should be useful in further elucidation of the transcriptional regulation mechanism of the gene and genetic analyses of type 2 diabetes. PMID- 19122347 TI - A brown tumor after biliopancreatic diversion for severe obesity. AB - A case of a brown tumor due to iatrogenic malabsorption following biliopancreatic diversion (BPD) is presented. A 52 year old women with a history of BPD 2 years before was referred to orthopedic surgery because of a painful lytic lesion of the left ankle. A bone biopsy revealed a giant cell tumor compatible with the diagnosis of a brown tumor. Subsequent metabolic evaluation showed severe 25 hydroxy vitamin D deficiency and secondary hyperparathyroidism (PTH 60 ng/L or twice the upper normal limit). Bone mineral density was decreased at the femoral neck (0.50 g/cm(2) ; T score of -3.92 or 66% of the expected value) and lumbar spine (T score of -1.75 or 93% of the expected value). A brown tumor can be the presenting symptom of iatrogenic malabsorption due to BPD. This case illustrates the severity of potential bone complications after BPD and the necessity of lifelong surveillance and vitamin supplements after BPD. PMID- 19122348 TI - Clinical significance of extrathyroid extension to the parathyroid gland of papillary thyroid carcinoma. AB - Extrathyroid extension is a prominent prognostic factor of papillary thyroid carcinoma (PTC). In the UICC TNM classification, minimal extension to the sternothyroid muscle and perithyroid soft tissue is classified as T3 and further massive extension is classified as T4, the highest T grade. However, there have been few studies on the clinical significance of extension to the parathyroid gland in a large case series. In this study, we investigated the prognosis of PTC with extension to the parathyroid gland in a series of 3208 patients who underwent initial surgery between 1997 and 2004. Of these patients, 51 (1.6%) showed extension to the parathyroid gland on pathological examination. Twenty-one of these patients had massive extrathyroid extension to other adjacent organs corresponding to pT4. The remaining 30 were enrolled in this study. The disease free survival (DFS) of these 30 patients was significantly better (p<0.0001) than that of pT4 patients and did not differ from that of patients showing minimal extrathyroid extension without extension to the parathyroid gland (p = 0.6264). Furthermore, none of these 30 patients died of carcinoma. Taken together, it is appropriate that extension to the parathyroid gland of PTC is graded as minimal extrathyroid extension (pT3), but not massive extension (pT4). Since minimal extension did not affect patient prognosis in our series, it is suggested that extension to the parathyroid gland has little clinical significance in PTC. PMID- 19122349 TI - Angiotensin II type 1 receptor blocker combined with calcium channel blocker for the treatment of obese hypertensive patients. PMID- 19122350 TI - Rapid identification of bacterial species with bacterial DNA microarray in cirrhotic patients with spontaneous bacterial peritonitis. AB - BACKGROUND/AIMS: Early detection and identification of bacteria in ascitic fluid could result in more timely treatment of cirrhotic patients with spontaneous bacterial peritonitis (SBP) or subclinical SBP. The aim of this study was to evaluate the usefulness of a bacterial DNA microarray for the rapid diagnosis of SBP and rapid bacterial identification in cirrhotic patients with ascites. METHODS: Thirty-seven cirrhotic patients with ascites (25 men and 12 women) participated. Ascitic fluid obtained from patients was tested by the bacterial DNA microarray method and by the conventional culture method. RESULTS: SBP and bacterascites were diagnosed in 8 (16.7%) of 48 specimens by the conventional method. The bacterial DNA microarray proved the existence of bacteria in 6 (75%) of 8 samples with SBP or bacterascites using the conventional method as a gold standard. A corresponding rate of bacterial species identification between the two methods was found in 5 of 6 samples (83.3%). It took 1.47+/-0.96 and 5.14+/ 2.6 days to receive the data by the microarray and conventional method, respectively (p<0.0001). After antibiotic therapy, the cumulative survival rate of recovered cases (n=8) was higher than that of unrecovered cases (n=5) (p=0.0008). CONCLUSION: Although the detection rate of the bacterial DNA microarray was similar to the conventional culture method, the DNA microarray could identify pathogens about 4 times more rapidly than bacterial cultivation, thus rendering it useful for managing cirrhotic patients with ascites. PMID- 19122351 TI - Nucleotide analogs for patients with HBV-related hepatocellular carcinoma increase the survival rate through improved liver function. AB - BACKGROUND AND AIM: This study evaluated the outcomes of antiviral therapy with nucleotide analogs for hepatitis B virus infection-related hepatocellular carcinoma. METHODS: Thirty patients orally received nucleotide analogs and, as a matched control group, 20 patients who were not treated with nucleotide analogs were selected. We compared changes in liver function, HCC recurrence and survival rate between both groups. RESULTS: In the nucleotide analog group, serum albumin, AST and ALT were significantly improved compared with baseline values. The Child Pugh score was significantly decreased in the nucleotide analog group. Furthermore, of the 36 patients curatively treated with the initial treatment, more patients in the nucleotide analog group improved or maintained their Child Pugh score at the time of recurrent HCC than in the control group (p=0.023). The cumulative recurrent-free survival rate of HCC did not significantly differ between the two groups; however, the cumulative survival rates of not only curative-treated patients but also all patients in the nucleotide analog group were significantly higher than those of patients in the control group (p=0.047 and p=0.02, respectively). CONCLUSION: The results suggest that nucleotide analog treatment increases the survival rate in patients with HCC by contributing to the improvement of remnant liver function. PMID- 19122352 TI - Effects of daily physical activity level on the degree of sympathetic tone. AB - BACKGROUND: Exercise is recommended for the treatment and prevention of cardiac disorders. Relaxation of the sympathetic nervous system has been considered to be one of the therapeutic mechanisms. However, the means by which the level of daily physical activity affects sympathetic activity remains unclear. OBJECTIVE: To evaluate the effects of daily physical activity on autonomic nervous tone. METHODS: Daily physical activity was measured for 5 or more days in 26 patients with various cardiac disorders of NYHA class 1 or 2 and in 6 healthy individuals using an intelligent calorie counter. Recumbent resting ECG was recorded for 3 minutes immediately before waking up and after going to bed using a digital ECG. Low- (Lf, 0.03-0.15 Hz) and high- (Hf, 0.15-0.45 Hz) frequency components were calculated by frequency analysis of the R-R interval, and Lf/Hf ratio was calculated as an index of sympathetic tone. RESULTS: The average values of energy expenditure and time were 145+/-93.6 kcal/day and 47.8+/-24.3 min/day, respectively. The morning Lf/Hf ratio decreased following an increased physical activity level the day before, but increased with subsequent increase in the activity level in 65.6% of subjects. A negative correlation was observed in 34.4% of subjects, which suggested that an appropriate level of physical activity led to relaxation of sympathetic tone. CONCLUSION: The daily level of physical activity affects sympathetic tone, and an appropriate level results in sympathetic relaxation. The results of this study provide a useful index to enable patients with cardiac disorder to perform exercise without overloading. PMID- 19122353 TI - Improved cardio-respiratory fitness correlates with changes in the number and size of small dense LDL: randomized controlled trial with exercise training and dietary instruction. AB - OBJECTIVE: To investigate the correlation between lifestyle improvements, in particular increased cardio-respiratory fitness and changes in the blood lipid profile. METHODS: The participants were 217 residents of Fuji and Yamato Towns, Saga City, with mildly abnormal blood pressure, serum lipids or blood glucose detected at health check-ups in 2003. Participants were randomly allocated to an Intervention (108 subjects) or Control group (109 subjects), matched for age and various conditions. The Intervention group was given exercise advice and prescription and dietary instructions. Cardio-respiratory fitness was evaluated using the work rate at double product breaking point. Changes of lipid parameters were compared before and after intervention, and examined the relationship with cardio-respiratory fitness. RESULTS: Seventy nine subjects in each group could be followed up for 17 months without requiring pharmacotherapy. Body mass index, waist circumference, systolic blood pressure, homeostasis model assessment insulin resistance, and triglycerides were decreased in the Intervention group. Furthermore, apolipoprotein B levels were lower (p<0.05), and the ratio of LDL cholesterol to apolipoprotein B (LDL/ApoB) was higher (p<0.001). When all subjects were divided into 3 subgroups according to the degree of improvement in cardio-respiratory fitness, LDL/ApoB increased and apolipoprotein B decreased as the degree of improvement increased (p<0.05). CONCLUSION: Improvement was seen in atherosclerotic risk factors through lifestyle modification. In particular, improved cardiorespiratory fitness was associated with qualitative and quantitative changes in LDLs. PMID- 19122354 TI - Comparison of 1 mg and 2 mg overnight dexamethasone suppression tests for the screening of Cushing's syndrome in obese patients. AB - OBJECTIVE: Obesity is currently a major public health problem and one of the potential underlying causes of obesity in a minority of patients is Cushing's syndrome (CS). Traditionally, the gold standard screening test for CS is 1 mg dexamethasone overnight suppression test. However, it is known that obese subjects have high false positive results with this test. DESIGN: We have therefore compared the 1 mg and 2 mg overnight dexamethasone suppression tests in obese subjects. Patients whose serum cortisol after ODST was >50 nM underwent and a low-dose dexamethasone suppression test (LDDST); 24-hour urine cortisol was collected for basal urinary free cortisol (UFC). For positive results after overnight 1-mg dexamethasone suppression test we also performed the overnight 2 mg dexamethasone suppression test. PATIENTS: We prospectively evaluated 100 patients (22 men and 78 women, ranging in age from 17 to 73 years with a body mass index (BMI) >30 kg/m2 who had been referred to our hospital-affiliated endocrine clinic because of simple obesity. Suppression of serum cortisol to <50 nM (1.8 microg/dL) after dexamethasone administration was chosen as the cut-off point for normal suppression. MEASUREMENTS: Thyroid function tests, lipid profiles, homocysteine, antithyroglobulin, anti-thyroid peroxidase antibody levels, vitamin B12, folate levels, insulin resistance [by homeostasis model assessment (HOMA)] and 1.0 mg postdexamethasone (postdex) suppression cortisol levels were measured. RESULTS: We found an 8% false-positive rate in 1 mg overnight test and 2% in 2 mg overnight test (p=0.001). There was no correlation between the cortisol levels after ODST and other parameters. CONCLUSIONS: Our results indicate that the 2 mg overnight dexamethasone suppression test (ODST) is more convenient and accurate than 1-mg ODST as a screening test for excluding CS in subjects with simple obesity. PMID- 19122355 TI - Extent of emphysema on HRCT affects loss of fat-free mass and fat mass in COPD. AB - BACKGROUND: Although muscle loss is thought to be a prognostic factor in chronic obstructive pulmonary disease (COPD), its determinants remain unclear. AIM: To verify the hypothesis that fat-free mass (FFM) and fat mass (FM) are associated with the extent of emphysema in COPD patients. PATIENTS AND METHODS: A total of 112 stable, male current or ex-smokers with or without COPD attending a secondary care specialist COPD clinic were studied. FFM and FM were measured by bioelectrical impedance analysis. We also assessed the nutrition status, muscle strength by the handgrip test, exercise tolerance by the 6-minute walking test, airflow limitation and diffusion capacity, the extent of emphysema by high resolution CT scan, systemic inflammation status using C-reactive protein, and a lipid-related hormone (adiponectin). RESULTS: The FFM index (FFMI), which was defined as the FFM divided by the square of the body height, was significantly correlated with age, the total number of lymphocytes, handgrip strength, distance on 6-minute walking, airflow limitation, diffusion capacity, extent of emphysema, and C-reactive protein. On multivariate analysis, the FFMI was associated with handgrip strength and inversely correlated with the extent of emphysema. The FM index (FMI) was positively correlated with pack-years, and was inversely correlated with the extent of emphysema and concentrations of adiponectin. CONCLUSION: The extent of emphysema was correlated with skeletal muscle loss and also the FM. PMID- 19122356 TI - A clinical comparative study of piperacillin and sulbactam/ampicillin in patients with community-acquired bacterial pneumonia. AB - OBJECTIVE: To evaluate the clinical usefulness of piperacillin (4 g/day) therapy for community-acquired pneumonia compared to sulbactam/ampicillin (6 g/day). METHODS: A randomized prospective clinical study was conducted in patients with mild to severe community-acquired bacterial pneumonia. RESULTS: The overall clinical efficiency of piperacillin therapy (4 g/day) in these patients (41/53=77.4%) was comparable to that of sulbactam/ampicillin therapy (6 g/day: efficiency rate: 33/49=67.3%), when each therapy was administered intravenously for 3-7 days. With regards to clinical efficiency based on disease severity, bacteriological efficiency, improvement in chest X-ray findings and adverse reactions, the two therapies were comparable, even though we found more efficiency for patients who had underlying diseases and there were also cost benefits in piperacillin therapy, compared with sulbactam/ampicillin therapy CONCLUSION: The results suggested that piperacillin therapy has good efficiency and tolerability and that it may be highly effective, even in cases of pneumonia with underlying diseases. This regimen may thus serve as a first line treatment of community-acquired pneumonia. PMID- 19122357 TI - Late onset neutropenia and immunoglobulin suppression of the patients with malignant lymphoma following autologous stem cell transplantation with rituximab. AB - BACKGROUND: Recently, decrease of immunoglobulin concentrations or neutrophil counts were reported in some cases several months after administration of rituximab. We examined a number of episodes of late onset neutropenia or immunoglobulin decrease in patients with malignant lymphoma who were in complete remission following autologous transplantation with or without rituximab. METHOD: Patients with follicular lymphoma and diffuse large B cell lymphoma transplanted with or without rituximab in our institutes were analyzed. Immunoglobulin concentrations and neutrophil counts after transplantation, both with and without rituximab were measured serially. RESULTS: Four weeks after transplantation, blood samples revealed lower concentrations of IgG and IgA in the rituximab group than in the non-rituximab group. Neutrophil numbers did not fall below 0.5x10(9) /L four weeks or more after transplantation in the non-rituximab group. Neutrophil numbers dropped below 0.5x10(9) /L in 6 of 14 cases in the rituximab group. CONCLUSION: Although the present study was retrospective and disease composition and pre-transplantation regimens differed between the two groups, the addition of rituximab to autologous transplantation might bring about a decrease of immunoglobulin levels and transient LON. PMID- 19122358 TI - Intramedullary cervical spinal cord abscess by viridans group Streptococcus secondary to infective endocarditis and facilitated by previous local radiotherapy. AB - The risk factors, microbial patterns, and prognosis of intramedullary abscess have varied with time. The development of an intramedullary abscess of the spinal cord (IASC) constitutes an exceptional complication of infective endocarditis (IE) in the post-antibiotic era. We present a case of cervical IASC by viridans group Streptococcus in a patient with mitral valve IE. We hypothesize that previous cervical radiotherapy for non-Hodgkin's lymphoma favoured the occurrence of this uncommon entity. This physiopathologic mechanism has not been previously reported. PMID- 19122359 TI - Causal relationships between esophageal squamous cell carcinoma and nephrotic syndrome. AB - We present a case of membranous nephropathy (MN) associated with esophageal squamous cell carcinoma (ESCC) with a high serum level of squamous cell carcinoma antigen (SCC). ESCC reached complete response (CR) for radiotherapy, with a partial remission of the proteinuria and decreased serum SCC levels. Six months after radiotherapy, the ESCC recurred, and the patient was treated with endoscopic mucosal resection (EMR), achieving a pathologic CR and disappearance of proteinuria and normalized serum SCC levels. The correlation of proteinuria and the serum level of SCC indicates that SCC could be a pathogenic antigen, responsible for the pathogenesis of MN in this patient. PMID- 19122360 TI - Cerebral infarction associated with heparin-induced thrombocytopenia in a patient with encephalitis. AB - We report a patient who had cerebral infarction associated with heparin-induced thrombocytopenia (HIT) during treatment of aseptic encephalitis. In patients with intracranial inflammation, such as ours, the possibility of HIT has to be considered when heparin is used, since inflammatory cerebral lesions often cause vascular damage, which is an aggravating factor for HIT-associated thrombosis. PMID- 19122361 TI - Pure dysarthria resulting from a small cortical infarction located at the left middle frontal gyrus. AB - Pure dysarthria caused by a small cortical infarction is rare. Recent reports have revealed that a small cortical lesion can lead to pure dysarthria, furthermore some reports have revealed that a middle frontal gyrus lesion might cause pure dysarthria. We report a 64-year-old woman who presented only dysarthria and had a small limited cortical infarction located at the left middle frontal gyrus. This case indicated that an isolated middle frontal gyrus lesion can cause pure dysarthria by secondary compromising of the cortical areas related or connected to the corticobulbar tract. PMID- 19122362 TI - Cardiac papillary fibroelastoma as a cause of embolic stroke: ultrasound and histopathological characteristics. AB - Although a cardiac papillary fibroelastoma is a benign cardiac tumor, it can cause severe embolic complications. A 51-year-old man presented with an ischemic stroke in the right middle cerebral artery territory. Transesophageal echocardiography revealed a small mobile tumor on the mitral valve as the only detectable source of emboli to the brain. On histology, the tumor was diagnosed as a papillary fibroelastoma. In this paper, the detailed characteristics of the tumor on ultrasound and histopathology are documented. In patients with cryptogenic stroke, transesophageal echocardiography should be done to rule out such an unusual emboligenic heart disease. PMID- 19122363 TI - Rectus sheath hematoma. PMID- 19122364 TI - Cardiac Erdheim-Chester. PMID- 19122365 TI - Neuronal injury induces cytokine-induced neutrophil chemoattractant-1 (CINC-1) production in astrocytes. AB - Accumulating evidence indicates a pivotal role for neuroinflammation in ischemic and excitotoxic brain injury. Cytokine-induced neutrophil chemoattractant-1 (CINC 1) is a CXC chemokine implicated in the infiltration of inflammatory cells into the brain parenchyma. In this study, we investigated the effect of N-methyl-D aspartate (NMDA)-induced neuronal injury on CINC-1 production in the organotypic cortico-striatal slice cultures. Treatment with 50 microM NMDA for 3 - 4 h caused devastating neuronal damage and increased CINC-1 production. Immunohistochemical analysis revealed that the CINC-1 immunoreactivity was predominantly detected in astrocytes. NMDA failed to induce CINC-1 production in enriched astrocyte cultures or neuron-depleted slice cultures, suggesting that NMDA acted on neuronal cells to induce astrocytic CINC-1 production. NMDA-induced CINC-1 mRNA expression was significantly inhibited by U0126, a mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor. These results suggest that NMDA-evoked neuronal injury induced astrocytic CINC-1 production via a MEK/ERK signaling pathway. Manipulation of this signaling pathway may serve as a target for suppressing neuroinflammation and, thereby, treating ischemic brain injury. PMID- 19122366 TI - Expression and functional characterization of choline transporter in human keratinocytes. AB - Choline is essential for synthesis of the major membrane phospholipid phosphatidylcholine. Moreover, it serves as a precursor for synthesis of the neurotransmitter acetylcholine (ACh). Keratinocytes of the epidermis synthesize and release ACh. The uptake of choline is the rate-limiting step in both ACh synthesis and choline phospholipid metabolism, and it is a prerequisite for keratinocyte proliferation. However, the nature of the choline transport system in keratinocytes is poorly understood. In this study, we examined the molecular and functional characterization of choline uptake into cultured human keratinocytes. Choline uptake into keratinocytes was independent of extracellular Na(+), saturable, and mediated by a single transport system with an apparent Michaelis-Menten constant of 12.3 muM. Choline uptake was reduced when the keratinocyte membrane potential was depolarized by high K(+). These results provide evidence that the choline transport activity is potential-sensitive. Various organic cations inhibit the choline transport system. RT-PCR demonstrated that keratinocytes expressed mRNA for choline transporter-like protein 1 (CTL1), mainly the CTL1a subtype. The present biochemical and pharmacological data suggest that CTL1a is functionally expressed in human keratinocytes and is responsible for the uptake of choline and organic cations in these cells. PMID- 19122367 TI - Serofendic acid promotes stellation induced by cAMP and cGMP analogs in cultured cortical astrocytes. AB - We investigated the effect of serofendic acid, a neuroprotective substance derived from fetal calf serum, on the morphological changes in cultured cortical astrocytes. Cultured astrocytes developed a stellate morphology with several processes following exposure to dibutylyl cAMP (dbcAMP), a membrane-permeable cAMP analog; 8-Br-cGMP, a membrane-permeable cGMP analog; or phorbol-12-myristate 13-acetate (PMA), a protein kinase C activator. Serofendic acid significantly accelerated the stellation induced by dbcAMP- and 8-Br-cGMP. In contrast, the PMA induced stellation was not affected by serofendic acid. Next, we attempted to elucidate the mechanism underlying the dbcAMP-induced stellation and explore the site of action of serofendic acid. Both the stellation induced by dbcAMP and the promotional effect of serofendic acid were partially inhibited by KT5720, a specific protein kinase A (PKA) inhibitor. Furthermore, serofendic acid failed to facilitate the stellation induced by Y-27632, an inhibitor of Rho-associated kinase (ROCK). These results indicate that serofendic acid promotes dbcAMP- and 8 Br-cGMP-induced stellation and the promotional effect on dbcAMP-induced stellation is mediated at least partly by the regulation of PKA activity and not by controlling ROCK activity. PMID- 19122368 TI - Activation of c-Jun N-terminal kinase cascades is involved in part of the neuronal degeneration induced by trimethyltin in cortical neurons of mice. AB - The organotin trimethyltin (TMT) is known to cause neuronal degeneration in the central nervous system. A systemic injection of TMT produced neuronal damage in the cerebral frontal cortex of mice. To elucidate the mechanism(s) underlying the toxicity of TMT toward neurons, we prepared primary cultures of neurons from the cerebral cortex of mouse embryos for use in this study. Microscopic observations revealed that a continuous exposure to TMT produced neuronal damage with nuclear condensation in an incubation time-dependent manner up to 48 h. The neuronal damage induced by TMT was not blocked by N-methyl-D-aspartate receptor channel blocker MK-801. The exposure to TMT produced an elevation of the phosphorylation level of c-Jun N-terminal kinase (JNK)(p46), but not JNK(p54), prior to neuronal death. Under the same conditions, a significant elevation was seen in the phosphorylation level of stress-activated protein kinase 1, which activates JNKs. Furthermore, TMT enhanced the expression and phosphorylation of c-Jun during a continuous exposure. The JNK inhibitor SP600125 was effective in significantly but only partially attenuating the TMT-induced nuclear condensation and accumulation of lactate dehydrogenase in the culture medium. Taken together, our data suggest that the neuronal damage induced by TMT was independent of excitotoxicity but that at least some of it was dependent on the JNK cascades in primary cultures of cortical neurons. PMID- 19122369 TI - The antinociceptive effects of midazolam on three different types of nociception in mice. AB - Antinociceptive effects of systemically administered midazolam remain controversial. The present study was performed to investigate its antinociceptive effects on different types of nociception in mice. Four different doses of midazolam (1, 3, 10, and 30 mg/kg) were administered intraperitoneally (i.p.). Saline was used as a control. The hot plate test, tail pressure test, acetic acid writhing test, the running wheel test, and the balance beam test were performed following the drug administration. In the hot plate test and tail pressure test, i.p. midazolam produced significant antinociceptive effects with the 50% effective dose (ED(50)) of 3.46 mg/kg [confidence interval (CI), 1.99 - 6.01 mg/kg] and 3.52 mg/kg (CI, 2.77 - 4.47 mg/kg), respectively. In the acetic acid writhing test, i.p. midazolam also produced significant antinociceptive effects. In the running wheel test, no mice stopped running after saline or midazolam at 1, 3, or 10 mg/kg, but all mice stopped running 30 and 45 min after i.p. administration of midazolam at 30 mg/kg. In the balance beam test, 30 min after i.p. administration of saline or midazolam at 1, 3, and 10 mg/kg, all mice were able to stay on the beam for 90 s, none of them could with midazolam at 30 mg/kg. In conclusion, systemically administered midazolam had antinociceptive effects on acute thermal, acute mechanical, and acute inflammatory-induced nociception in mice. The antinociceptive potency of midazolam was the same for both acute thermal-induced nociception and mechanical-induced nociception. PMID- 19122370 TI - In vitro assessment of progesterone and prostaglandin e(2) production by the corpus luteum in cattle following pharmacological synchronization of estrus. AB - We studied the secretory function of the corpus luteum (CL) in cows following different estrus synchronization protocols. Estrus was synchronized using one (n=4) or two injections (n=5) of prostaglandin F(2alpha) (PGF(2alpha); dinoprost), two injections of different analogues of PGF(2alpha) (aPGF(2alpha)), luprostiol (n=5) and cloprostenol (n=5), at eleven-day intervals, a gestagen implant (norgestomet, n=5, for 10 days) or norgestomet together with a subsequent dinoprost injection on the day of implant removal (n=5). CL samples were collected by ovariectomy on Day 7-8 of the estrous cycle. Luteal strips were stimulated with LH (100 ng/ml) or prostaglandin E(2) (PGE(2), 10(-6)M) for 24 h in culture media. The progesterone (P(4)) and PGE(2) concentrations in the media were measured by enzyme immunoassay. In the control CL (spontaneous estrus; n=5), LH and PGE(2) stimulated P(4) and PGE(2) (P<0.001). The effects of both factors on P(4) were reduced in the CL following dinoprost- and cloprostenol-synchronized estrus (P<0.05) and were absent in the luprostiol-synchronized CL (P>0.05). In the norgestomet-synchronized CL, the stimulatory effects of LH and PGE(2) were higher compared with the CL synchronized by aPGF(2alpha) (P<0.05). Pharmacological manipulation of the estrous cycle using aPGF(2alpha) may cause lower P(4) secretion. Estrus synchronization inhibited CL sensitivity to luteotropic factors. Therefore, attention should be focused on the estrous synchronization method in both in vivo and in vitro studies of CL functions in cattle. PMID- 19122371 TI - Effect of synchronisation of ovulation on ovarian profile and days open in holstein cows diagnosed as nonpregnant 26 days after timed artificial insemination. AB - The objective of this study was to clarify the effects of a second protocol of ovulation synchronisation starting on day 26 after timed artificial insemination on ovarian profile and days open in dairy cows diagnosed as nonpregnant. Ninety four Holstein-Friesian cows received intramuscular injections of a GnRH analogue (GnRH), 100 microg fertirelin, on day 0 and a prostaglandin F(2alpha) analogue (PG), 5 mg etyprostontromethamine, on day 7. GnRH was again administered 48 h after the PG injection, and timed artificial insemination was performed 16 to 20 h later (Ovsynch/TAI). Twenty-six of the 94 cows returned to oestrus within 26 days after TAI and were inseminated. Of the other 68 cows, 44 were not pregnant and were randomly allocated to undergo another Ovsynch/TAI protocol (Resynch group; n=23) or AI only after detection of oestrus (Control group; n=21). The ovarian and hormonal profiles were compared between the first and second Ovsynch protocol periods in the Resynch group. The diameter of the dominant follicle and plasma oestradiol-17 beta concentration at the second GnRH injection were significantly greater than those at PG injection during the second Ovsynch period. Ovulation was synchronised in all of the animals in the second Ovsynch period. The AI submission rates, mean AI intervals and pregnancy rates of the Resynch and Control groups were 100% and 57.1%, 36.0 +/- 0.0 and 43.2 +/- 10.9 and 30.4% and 14.3%, respectively. The mean AI interval was 7 days shorter and the pregnancy rate was higher in the Resynch group than in the Control group, although no significant differences were found due to the small number of the animals. In conclusion, the Resynch protocol initiated on day 26 after TAI in the first protocol has the potential to reduce days open and increase the pregnancy rate in dairy cows. PMID- 19122372 TI - [Treatment of pulmonary hypertension associated with connective tissue disease]. AB - Pulmonary hypertension is the most ominous complication of connective tissue diseases and its treatment has been a big challenge to clinicians. Vasodilators including epoprostenol (prostacyclin), bosentan (endothelin-receptor antagonist), and a sildenafil (phosphodiesterase type 5 inhibitor) have recently become available in Japan. These vasodilators may improve exercise tolerance, hemodynamics, activities of daily life, and the life span. In this article, we review an endothelin-receptor antagonist and phosphodiesterase type 5 inhibitor recently approved as treatment modalities for pulmonary hypertension in Japan. PMID- 19122373 TI - [Antigen specific treatment for the inhibition and remission of type 1 diabetes]. AB - Treatment with anti-CD3 antibodies appears promising to preserve residual beta cell function in recent onset type 1 diabetes although many patients had therapy related adverse events. Insulin is an important islet antigen and autoimmunity to insulin may be central to disease pathogenesis of type 1 diabetes in man and NOD mouse. Evidence is strongest for the NOD mouse model, where blocking immune responses to insulin by amino acid substitution at positions B: 16, prevents diabetes. Insulin can be used to immunologically prevent diabetes of NOD mice, however, insulin-based preventive immunoregulation of diabetes in man is not yet possible. Treatment of NOD mice with insulin B-chain peptide and poly I: C, a Toll-like receptor 3 ligand, induces the pathogenic T cells as well as regulatory T cells and recruits them into the islets. Intranasal treatment with insulin B chain analogue peptide with amino acid substitutions at positions B: 16 and 19 prevented the progression to diabetes and induced remission from hyperglycemia when co-administered with a mucosal adjuvant cholera toxin. Thus, an antigenic peptide vaccination with an alternative adjuvant or route might induce antigen specific regulatory cell populations rather than pathogenic T cells. We believe that such an improved antigen specific therapy could provide more efficient and safer disease suppression and remission for human type 1 diabetes. PMID- 19122374 TI - [Sh2b3/Lnk family adaptor proteins in the regulation of lymphohematopoiesis]. AB - Sh2b3/Lnk consisting of an N-terminal proline-rich region, PH-, SH2-domains and a tyrosine phosphorylation site, forms an intracellular adaptor protein family conserved from drosophila to mammals, together with Sh2b1/SH2-B and Sh2b2/APS (adaptor protein with PH and SH2 domains). Lnk negatively regulates lymphopoiesis and early hematopoiesis. The lnk-deficiency results in enhanced production of B cells, and expansion as well as enhanced function of hematopoietic stem cells (HSCs), demonstrating negative regulatory functions of Sh2b3/Lnk in cytokine signaling. Our recent studies also revealed that Sh2b3/Lnk functions in responses controlled by cell adhesion and in crosstalk between integrin- and cytokine mediated signaling. Importantly, recent genome-wide association studies of the autoimmune type 1 diabetes or celiac disease identified risk variants in the SH2B3/LNK region, indicating possible unrevealed functions mediated by this adaptor molecule. This review summarizes roles of Sh2b3/Lnk in the regulation of B-lymphopoiesis and HSCs expansion and function, and briefly introduces our approach for modulating HSCs function by targeting Sh2b3/Lnk-mediated pathways. PMID- 19122375 TI - [Various clinical symptoms in human parvovirus B19 infection]. AB - Human parvovirus B19 infection causes erythema infectiosum in child, aplastic crisis in patients with chronic hemolytic anemia, chronic pure red cell aplasia in immunocompromised patients and hydrops fetalis. Human parvovirus B19 causes arthritis and acute glomerulonephritis due to immunological mechanism. Other disorders, rheumatoid arthritis, vasculitis and thrombotic microangiopathy, are linked in human parvovirus B19 infection. Parvovirus B19 infection causes choronic rheumatoid-like arthropathy. Autoantibody and low complement were seen in acute human parvovirus infection, and parvovirus B19 infection present clinically lupus like tableau. PMID- 19122376 TI - [Infliximab for a girl with refractory pyoderma gangrenosum]. AB - Pyoderma gangrenosum is a rare chronic ulcerative noninfectious disease of the skin. Half of patients are complicated with other autoimmune diseases, most commonly inflammatory bowel disease, Takayasu disease, and rheumatoid arthritis. It has been reported that approximately 4% of them were childhood-onset. The conventional treatments of pyoderma gangrenosum were described as systemic corticosteroids and cyclosporine. The combination of corticosteroids with immunosuppressants such as tacrolimus, mycophenolate mofetil has been reported as steroid-sparing modalities. We herein reported a girl, 12 years of age, having pyoderma gangrenosum refractory to the conventional combination of systemic prednisolone with cyclosporine, but successfully treated with infliximab, the anti-TNFalpha monoclonal antibody. Rapid improvement of pyoderma gangrenosum was seen within three doses of infliximab infusion. All skin lesions eventually healed completely and new skin ulcers were never coming out again. The dramatical improvement suggested that infliximab should be considered for patients with refractory pyoderma gangrenosum though further experiences and investigations are required to determine the mechanism of infliximab. PMID- 19122377 TI - [Successful treatment of early intervention with tacrolimus for a patient with lupus nephritis III+V]. AB - A 55-year-old woman developed bilateral leg edema in June 2006. Since the edema tended to worsen, she visited our hospital on November 11. Laboratory tests showed a serum albumin level of 2.5 g/dl with 3+ proteinuria, and suggested nephrotic syndrome, which led to her hospitalization on November 14. The findings of discoid erythema, an antinuclear antibody titer of 1: 640, anti-ds DNA antibody titer of 16.8 IU/ml, and ISN/RPS class III (A/C)+V lupus nephritis on kidney biopsy led to the diagnosis of systemic lupus erythematosus. Treatment with prednisolone at 1 mg/kg/day was initiated. Despite an increase in complement levels and decreases in anti-ds DNA antibody titers and immune complexes, proteinuria persisted; therefore, the patient was concomitantly given 125 mg/day of azathioprine, which was discontinued because of the poor improvement of proteinuria and development of myelosuppression, and replaced by 3 mg/day of tacrolimus (Tac), with a consequent marked improvement in proteinuria. We report a patient with refractory proteinuria due to class III/V lupus nephritis who achieved a Tac-induced complete remission. PMID- 19122378 TI - [A case of progressive multifocal leukoencephalopathy associated with systemic lupus erythematosus]. AB - We describe a case of systemic lupus erythematosus (SLE) complicated with multifocal leukoencephalopathy (PML). A 57-year old woman, who had a five-year history of SLE, was admitted to our hospital because of fever and multiple subcutaneous nodules. Diagnosis of disseminated cryptococcosis was made based on histological and bacteriological examinations, and she was successfully treated with anti-fungal drugs. Corticoteroids were increased for persistent lupus activities. One month later, however, she gradually developed disorientation and short-term memory loss. A brain magnetic resonance image (MRI) showed a focal lesion in the white matter of the right frontal lobe. Brain biopsy demonstrated demyelinating lesions with the presence of JC viral antigen. Polymerase chain reaction also revealed JC virus DNA in the cerebrospinal fluid. Her condition gradually progressed, and she died a year later due to pneumonia. Although acquired immunodeficiency syndrome is currently the most common disease associated with PML, patients with autoimmune diseases receiving immunosuppressive therapy also have risks for developing PML. In patients with SLE presenting with subacute neurological abnormalities and white matter lesions in the brain, PML should be considered in the differential diagnosis. PMID- 19122379 TI - Involvement of Na(+)-Ca (2+) exchanger in the endothelium-independent vasorelaxation induced by Curcuma longa L. in isolated rat superior mesenteric arteries. AB - Previous studies confirmed that the methanolic extract from Curcuma longa L. (CLME) lowers arterial blood pressure and heart rate in rats due to the blockade of extracellular Ca(2+) influx. The aim of this study was to investigate the involvement of Na(+)-Ca (2+) exchanger in the vasorelaxant effects elicited by CLME in isolated rat superior mesenteric arteries. CLME (1-1,000 microg/ml) concentration-dependently relaxed phenylephrine (PHE) (10 microM) pre-contracted arterial rings with intact-endothelium (pD(2) and E(max) = 2.04 +/- 0.06 and 88.3 +/- 3.2%) or denuded-endothelium (pD(2) and E(max) = 2.06 +/- 0.03 and 91.4 +/- 1.0%), respectively, suggesting that the removal of endothelium has no significant effect (P>0.05) on the vasorelaxation induced by CLME. Furthermore, CLME (30, 100 and 300 microg/ml) inhibited the cumulative concentration-response curves to PHE (10(-8)-10(-5) M) in a concentration-dependent manner, whereas, treatment with ouabain 100 microM (selective blocker of Na(+)-K(+) ATPase) has no effect on the relaxant responses of CLME. However, treatment with nickel chloride (NiCl(2)) (100, 300 and 400 microM), a putative Na(+)-Ca(2+) exchanger inhibitor, concentration-dependently reduced the vasorelaxant responses of CLME. Precisely, NiCl(2) at 100, 300 and 400 microM significantly (P<0.05) decreased the pD(2) and E(max) values of CLME (1.86 +/- 0.03 and 81.3 +/- 1.2%, 1.77 +/- 0.03 and 60.2 +/ 0.8%, 1.69 +/- 0.04 and 55.3 +/- 1.6%, respectively). Also, CLME (100 microg/ml) produced less relaxant effect with decreasing extracellular Na(+) concentration. CLME-induced vasorelaxation was completely abolished in a Na(+)-free Tyrode's solution, a condition that eliminates the influence of the forward mode of the exchanger. The results provide indirect evidence that the stimulation of the forward mode of Na(+)-Ca (2+) exchanger may probably contribute to the vasorelaxation induced by CLME in endothelium-denuded arterial rings. PMID- 19122380 TI - The utility of noninvasive (13)C-acetate breath test using a new solid test meal to measure gastric emptying in mice. AB - In clinical and experimental settings, the (13)C breath test is performed to measure gastric emptying and has advantages of noninvasiveness and repeatability. We intended to apply the (13)C breath test method to mice with an easy-to-handle solid test meal that is more physiological than liquid meals. Male ddY mice were trained to eat (13)C-acetate-containing pellets as the solid test meal. Thirty minutes after administration of metoclopramide (0.3-3 mg/kg, p.o.) or atropine sulfate (0.3-3 mg/kg, i.p.), mice received the test meal and were placed in chambers. The (13)CO(2) levels in the expired air were measured and the maximum concentration (C(max); per thousand) and the time to reach the maximum concentration (T(max); min) were determined. Metoclopramide significantly and dose-dependently increased C(max) and decreased T(max). On the other hand, atropine sulfate significantly and dose-dependently decreased C(max) and increased T(max). The (13)C-acetate breath test using a solid test meal is sensitive enough to detect both enhanced and delayed gastric emptying of the reference drugs. PMID- 19122381 TI - Uterolytic effect of Hypoxis hemerocallidea Fisch. & C.A. Mey. (Hypoxidaceae) corm [;African Potato'] aqueous extract. AB - Extracts of Hypoxis hemerocallidea corm (African potato) are commonly used by some traditional health practitioners in KwaZulu-Natal Province of South Africa as natural antenatal remedy to prevent threatening or premature abortion and miscarriage, and to ensure successful confinement. In this study, we investigated the uterolytic activity of H. hemerocallidea corm aqueous extract on spontaneous, rhythmic contractions of uterine horns taken from pregnant rats and guinea-pigs, as well as on spasmogen-provoked contractions of stilboesterol-primed, oestrogen dominated, non-pregnant rat and guinea-pig isolated uterine horns. Relatively low to high concentrations of H. hemerocallidea corm aqueous extract (APE, 25-400 mg/ml) inhibited the amplitude of the spontaneous, rhythmic contractions of, and relaxed, uterine horns isolated from pregnant rats and guinea-pigs in a concentration-related manner. Furthermore, relatively low to high concentrations of APE (25-400 mg/ml) relaxed basal tones of uterine horns taken from non pregnant, oestrogen-dominated rats and guinea-pigs in a concentration-dependent manner. The same moderately low to high concentrations of APE (25-400 mg/ml) inhibited acetylcholine-, oxytocin-, bradykinin-, and potassium chloride (K(+)) induced contractions of oestrogen-dominated rat and guinea-pig isolated uterine horns in a concentration-related manner. Although the mechanism of uterolytic action of APE could not be established, the results of the present study lend pharmacological credence to the folkloric, ethnomedical uses of APE as a natural antenatal remedy for threatening or premature abortion, and suggest that the uterolytic action of the corm's extract is unlikely to be mediated via beta(2) adrenoceptor stimulation, but probably mediated through a non-specific spasmolytic mechanism. PMID- 19122383 TI - X-ray crystallographic analysis of measles virus hemagglutinin. AB - X-ray crystallographic analyses, together with nuclear magnetic resonance, have revealed three-dimensional structures of many important viral proteins, thereby allowing us to better understand the interactions between viral and host cell molecules. In this review, we summarize the recently determined crystal structure of the measles virus (MV) attachment protein hemagglutinin. Based on this structural information, we also discuss how the MV hemagglutinin interacts with various cellular receptors and why MV vaccines have been effective for many years without inducing escape mutant viruses. Other topics discussed are a putative MV receptor present on polarized epithelial cells and the protein expression system using a cultured human cell line 293SGnTI(-), which is suitable for X-ray crystallographic analyses. PMID- 19122382 TI - Effect of long-term streptozotocin-induced diabetes on coronary vasoconstriction in isolated perfused rat heart. AB - The primary goal of this study was to investigate the effect of long-term (9 months) streptozotocin (STZ)-induced diabetes on the coronary vasoconstrictor responses to vasoactive agents such as high K(+), acetylcholine (ACh), endothelin 1 (ET-1), and the calcium-channel activator Bay K 8644. For this, we used isolated rat hearts perfused at constant flow rate. Each of the four agents caused dose-dependent increases in perfusion pressure in isolated hearts from age matched control and STZ-induced diabetic rats. The dose-response curves for high K(+), ACh, and ET-1 were shifted to the left, so that at some lower doses of these agents the increased perfusion pressure was greater in coronary arteries obtained from diabetic rats than in those from control rats. On the other hand, the maximum contractile response induced by each of these agents was lower in the diabetic perfused heart. The Bay K 8644-induced contractile response was significantly greater in the coronary arteries of diabetic rats than in those of control rats. A threshold-constrictor concentration of Bay K 8644 (1 nM) potentiated the ACh-induced vasoconstriction in coronary arteries from both groups, and the potentiated responses were greater in diabetic rats than in controls at lower concentrations of ACh (100 nM and 1 microM). These findings suggest that the coronary artery contractile responses to lower concentrations of ACh or ET-1 are exaggerated in long-term STZ-induced diabetic hearts. These changes may be due to alterations in the activity of voltage-gated Ca(2+) channels. PMID- 19122385 TI - HCV and innate immunity. AB - Hepatitis C virus (HCV) is a single-strand, positive sense RNA virus belonging to the flaviviridae family. HCV develops persistent infection in >70% of infected patients, and eventually causes chronic hepatitis, cirrhosis, and hepatocellular carcinoma in some patients. Once chronic infection is established in patients with HCV, spontaneous viral clearance fails, although how HCV remains persistently infecting the liver is largely unknown. Insufficient immune response, involving antiviral innate immune response including dendritic cells (DCs), has been focused. A number of controversial studies have been reported as to HCV genome replication and HCV-mediated immune responses in human DCs. A tantalizing point of these earlier studies is the lack of the system for viral propagation in HCV. Recently, an in vitro system was exploited to propagate HCV particles using the JFH1 strain. In this review, we review the previous reports about the subversion of innate immunity by HCV and show the innate response of monocyte-derived dendritic cells (MoDCs) against the JFH1 strain. We could not observe HCV direct interaction with MoDC maturation. MoDCs maturated by phagocytosing HCV-infected apoptotic cells containing virus-derived dsRNA, which interacted with TLR3 in the phagosomes. All of these data suggests the importance of TLR3 signal for the induction of anti-HCV innate immunity. PMID- 19122384 TI - Telomerase-specific oncolytic virotherapy for human cancer with the hTERT promoter. AB - Replication-selective tumor-specific viruses present a novel approach for treatment of neoplastic disease. Telomerase activation is considered to be a critical step in carcinogenesis and its activity correlates closely with human telomerase reverse transcriptase (hTERT) expression. We constructed an attenuated adenovirus 5 vector (Telomelysin, OBP-301), in which the hTERT promoter element drives expression of E1 genes. Telomelysin replicated efficiently and induced marked cell killing in a panel of human cancer cell lines, whereas replication as well as cytotoxicity was highly attenuated in normal human cells lacking telomerase activity. We further modified the E3 region of OBP-301 to contain green fluorescent protein (GFP) gene for monitoring viral replication (TelomeScan, OBP-401). When TelomeScan was intratumorally injected into human tumors orthotopically implanted into the rectum in mice, para-aortic lymph node metastasis could be visualized at laparotomy under a three-chip color cooled charged-coupled device camera. This article reviews recent highlights in this rapidly evolving field: cancer therapeutic and cancer diagnostic approaches using the telomerase-specific oncolytic adenoviruses. PMID- 19122386 TI - HSV-1 infection through inhibitory receptor, PILRalpha. AB - Paired receptors that consist of highly related activating and inhibitory receptors are widely involved in the regulation of immune response. Several viruses that persistently infect hosts possess genes that encode ligands for inhibitory receptors in order to escape from host immune system. Herpes simplex virus type 1 (HSV-1) is one of the viruses that cause persistent infection. Here, we found that HSV-1-infected cells express a ligand for paired immunoglobulin like-type 2 receptor (PILR)alpha, one of paired inhibitory receptors mainly expressed on myeloid cells such as monocytes, macrophages and dendritic cells. Furthermore, we have identified that glycoprotein B (gB), an envelope protein of HSV-1, is a ligand for PILRalpha by mass spectrometry analysis. Because gB is essential for HSV-1 to infect cells, we analyzed function of PILRalpha in HSV-1 infection. When PILRalpha was transfected into CHO-K1 cells, which is resistant to HSV-1 infection, the PILRalpha-transfected CHO-K1 cells became permissive to HSV-1 infection. We further addressed weather PILRalpha is involved in the HSV-1 infection of primary human cells. CD14-positive monocytes that express both PILRalpha and HVEM, a glycoprotein D receptor, were susceptible to HSV-1 infection. In contrast, HSV-1 did not infect CD14-negative lymphocytes that express HVEM but not PILRalpha. Furthermore, HSV-1 infection of monocyte was blocked by both anti-PILRalpha mAb and anti-HVEM antiserum. These findings indicated that both gB and gD receptors play an important role in HSV-1 infection. We have shown, for the first time, that viruses use an inhibitory immune receptor to enter a cell. Invasion into hematopoietic cells by using inhibitory receptors should be beneficial to the virus because binding to inhibitory receptors may not only provide entry, but also trigger the inhibitory receptor to suppress the immune functions of the infected cell. PMID- 19122387 TI - DNA sensors in innate immune system. AB - Microbial sensing mediated by pattern recognition receptors (PRRs) is the first key step to trigger innate immune responses, represented by the induction of type I interferons (IFNs), proinflammatory cytokines and chemokines. This innate signaling elicits an efficient activation of more specific responses in adaptive immunity. Such coordinated responses in the two systems are essential for the optimal elimination of invading microbes. Despite a major advance in our understanding of RNA sensors, TLR9 remained the only known sensor of DNA. On the other hand, there has been accumulating evidence supporting the existence of TLR9 independent DNA recognition mechanism. In this regard, DAI (also termed as DLM 1/ZBP1), the first sensor of cytosolic DNA, has recently been identified with its activation of IFN-regulatory factors(IRFs) and NF-kappaB transcriptional factors. Several recent papers suggest the involvement of an additional cytosolic DNA sensor(s). There is also a recent report that cytosolic microbial and host DNA can trigger pro-inflammatory responses via the TLR- and IRF-indepnedent pathway mediated by the inflammasome, which is consisted of NLR family members together with the adaptor protein ASC and caspase-1. In addition, evidence has been provided that host- and virus-derived proteins, which contain DNA-binding motifs (Zalpha and/or Zbeta) similar to those of DAI(DLM-1/ZBP1), negatively regulates the immune response that is activated by cytosolic DNA. Thus, these recent findings reveal the complex DNA-sensing mechanism for triggering the activation of innate immunity, and the breakdown of this sensing mechanism may lead to autoimmune abnormalities. PMID- 19122388 TI - Regulation of viral recognition signaling by ubiquitin modification. AB - As a defense mechanism against infection, host cells have evolved sensor molecules which detect pathogen components directly and induce protective responses against the infection. TLRs, well known receptors, recognize a pathogen on the surface of cells or endosome/lysosome. Many pathogens penetrate into cytoplasm, in where non-TLR sensors recognize pathogen components including double-stranded RNA (dsRNA). On the downstream of each sensor, a variety of functional signaling molecules are activated to produce various cytokines upon the microbial invasion to induce host defense responses. Because that cytokines produced to regulate the host defense responses are known to affect cell proliferation also, the level of these molecules are needed to be controlled tightly, which means requisites of negative regulation of the signaling activated by pathogen after the completion of proper immune responses. Recent studies suggest important roles of some ubiquitin systems in this regulation. Here we focus, in particular, ubiquitin conjugation to signaling molecules by virus activation and like to show how ubiquitin signaling plays roles in this regulation by introducing some recent works. PMID- 19122389 TI - Transposable elements, RNA silencing, and their impacts on the genome throughout evolution. AB - It is remarkable to consider that more than 40% of the human genome is comprised of transposable elements (TEs) and their relics. TEs were long thought of as either 'selfish' or 'parasitic' DNA elements that were there not for the sake of the host organism, but for their own sake in an evolutionary sense; thus they were considered to be either neutral or deleterious to their hosts. However, it is becoming increasingly clear that there are more complex interactions between TEs and their hosts than strict parasitism; these elements produce changes that have a broad range of fitness values at an organismal level. Recent evidence indicates that these elements confer a fitness benefit to the host more frequently than previously recognized. RNA silencing is thought to have evolved as a form of nucleic-acid-based, and thus sequence-directed, immunity to block the action of viruses and TEs. Host-parasite interactions are typically associated with rapid evolution because of a permanent antagonistic relationship resembling an "arms race" in which parasite adaptations are countered by host adaptations. Complex interactions between TEs and RNA silencing machineries have been co-opted to regulate cellular genes. PMID- 19122390 TI - Viruses and RNA silencing. AB - Small RNAs play a critical role in the regulation of gene expression in diverse cellular processes. This mechanism, termed RNA silencing or RNAi, also functions as a defense mechanism against molecular parasites such as virus and transposon. Whereas RNA silencing is triggered by viral infection, viruses suppress RNA silencing to establish infection, and sometimes even exploit it for their infection. In this mini review, we describe intimate interactions between viruses and host organisms in RNA silencing. PMID- 19122391 TI - Host adaptation mechanisms of Influenza A viruses. AB - Influenza A viruses are a member of Orthomyxoviridae possessing negative-sense, single-stranded, and 8 segmented RNA genomes. These 8 RNA segments encode individual specific viral proteins, and up until now a total of 11 proteins have been identified including alternate open reading frame in one genome. Because of this simplicity, utilization of many host systems is required during the viral infection cycle in a cell. Therefore adaptation to the host cell system is the most important issue for the influenza A virus infection and transmission. Although there are many factors which relate to the host adaptation in the viruses themselves, elucidation of mechanisms about changing viral receptor specificities is one of the most advanced research areas. Because the avian type viral receptor was found in the human lungs, the situation, such as co-existence of avian- and human-derived viral receptors in the human airway, may contribute to the human adaptation of avian derived influenza A viruses. PMID- 19122392 TI - Chemical biology for revealing a life cycle of hepatitis C virus. AB - It is one of the major subjects in the virology field to develop novel anti-viral strategies as well as to reveal the mechanism of viral replication. We have developed a method of chemical compound-directed analysis on viral replication. Until now, we rediscovered cyclosporin A and tamoxifen as anti-hepatitis C virus (HCV) compounds. Through analysis using these compounds, it was revealed that host cell factors cyclophilin (CyP) and estrogen receptor were important for HCV replication. CyP was demonstrated to serve as a new target of the development for anti-HCV agents and CyP inhibitors are now under clinical trials. Thus, an application of chemical biology to virology would provide not only mechanistic aspects of viral life cycles but also new anti-viral strategies. PMID- 19122393 TI - Localization of sympathetic, parasympathetic and sensory neurons innervating the distal ileum of the cattle. AB - After oral challenge of the pathological prion protein, the causative agent of bovine spongiform encephalopathy, the pathogen was first detected in the distal ileum and then deposited in the brain. The present study aims determining the possible neuronal transporting pathways from the ileum to the brain in the cattle using a tracer protein. After horseradish peroxidase was injected into the wall of the distal ileum in the calf, almost all labeled neurons were detected in the celiac and cranial mesenteric ganglion complex. Only a few labeled neurons existed in the caudal mesenteric ganglion and the paravertebral ganglia. They were sympathetic postganglionic neurons. In the dorsal root ganglia T5 to L4, some sensory neurons were found to be labeled. Only a small number of parasympathetic preganglionic neurons were labeled in the dorsal motor nucleus of the vagus nerve. No labeled sensory neurons were found in the nodose ganglion. These results suggest that the pathological prion protein is mainly transported to the spinal cord and brain via the sympathetic nervous system and partially via the sensory neurons in the dorsal root ganglia. The vagus nerve does not seem to contribute to the transport of the pathogen from the ileum directly. PMID- 19122394 TI - Effect of collagen oligopeptide injection on rabbit tenositis. AB - Effects of the collagen oligopeptide (COP) were examined by its repeat injection into the inflammatory rabbit Achilles tendon (Tendo calcaneus communis), in which tenositis was induced by injection of bacterial collagenase. COP was evaluated 5 times over a period of 3 weeks to 1 month after injection of collagenase. At 1 month after treatment, the therapeutic effect of COP was evaluated by examining the structure of collagen fibrils, amount and components of glycosaminoglycans (GAGs) and matrix metalloproteinases (MMPs), and compared with the saline injection, control, and normal groups. The Achilles tendon of rabbit in the control group (no COP injection) and saline injection group showed a notable increase in the number of fine collagen fibrils, a change in GAG composition and increase in the amount of pro-MMP-2, indicating the weakening of the tendon. In contrast, the size distribution of collagen fibrils, GAG composition and the amount of pro-MMP-2 was similar to those in the normal group. These results suggest that COP injection promotes healing processes of the tendon injury. PMID- 19122395 TI - High seroprevalence of Encephalitozoon cuniculi in pet rabbits in Japan. AB - Infection with Encephalitozoon cuniculi in rabbits frequently exists as a chronic, latent infection, and only a percentage of infected animals develop clinical disease. This study presents a seroepidemiological study of E. cunicucli infection in 337 pet rabbits collected from 20 prefectures in Japan in 2006 and 2007, using enzyme-linked immunosorbent assay (ELISA) capable of measuring IgG and IgM antibodies. These rabbits were divided into the following four groups: healthy and isolated rabbits (n=74, group I), healthy and companioned rabbits (n=121, group II), neurologically diseased rabbits (n=105, group III), and other diseased rabbits (n=37, group IV). Using ELISA for IgG antibodies, the highest detection rate, 81%, was seen in group III, the second highest, 75.2%, in group II, and the lowest, 29.7%, in group I, which was significantly different to the other groups except for group IV (43.2%). On the other hand, when ELISA was used for IgM antibody detection, 14-40% of rabbits in the four groups were also observed to have anti-E. cuniculi IgM. This study demonstrated high seroprevalence of E. cuniculi in not only neurologically diseased rabbits but also healthy and other diseased rabbits. PMID- 19122396 TI - Recombination between vaccine and field strains of canine parvovirus is revealed by isolation of virus in canine and feline cell cultures. AB - Canine parvovirus type 2 (CPV) is a pathogen that causes severe hemorrhagic gastroenteritis with a high fatality rate in pups worldwide. Since CPV emerged in the late 1970s, its origin has been explored with the conclusion that CPV originated from feline panleukopenia virus or a closely related virus. Both high mutation rate and recombination are assumed to be key factors in the evolution of parvoviruses. Here we provide evidence for natural recombination in CPV isolated from dogs in cell culture. Antigenic and genetic properties of isolates from 10 diseased pups were elucidated. Six pups had been vaccinated beforehand with live combined vaccine containing original antigenic type CPV (CPV-2). Six isolates recovered from 4 vaccinated pups in cell cultures were found to contain either CPV-2 or CPV-2-like viruses. The other isolates, including all those from non vaccinated pups, were CPV-2b viruses. Antigenic typing of two CPV-2-like isolates, 03-029/M and 1887/f, with a monoclonal antibody panel suggested they were a mixture of CPV-2 and CPV-2a (03-029/M) and a recombinant of CPV-2 and CPV 2b (1887/f). Genetic analysis of the VP1 gene indicated that isolate 03-029/M was a mixture of CPV-2, CPV-2a and a recombinant of CPV-2 and CPV-2a viruses, while isolate 1887/f was composed of a recombinant of CPV-2 and CPV-2b viruses. This is the first demonstration of natural CPV recombination in the field and suggests that recombination in the evolution of CPV is a more frequent and important process than previously believed. PMID- 19122397 TI - Antibody-dependent enhancement occurs upon re-infection with the identical serotype virus in feline infectious peritonitis virus infection. AB - Feline infectious peritonitis virus (FIPV) is classified into serotype I and serotype II according to the amino acid sequence of its spike(S) protein. Antibody dependent enhancement (ADE) of macrophage infection occurs in the presence of antibodies to FIPV S protein, and a close relationship between ADE and neutralizing epitopes has been reported. The importance of differences in FIPV serotype on the induction of ADE remains unclear. In this study, we investigated whether the same or different serotype of FIPV induces ADE in cats. Specific pathogen-free cats were passively immunized with anti-type I or II FIPV antibodies, and we investigated the induction of ADE following subcutaneous inoculation with type I FIPV. Inoculation using FIPV serotype I enhanced the onset of FIP in cats passively immunized with FIPV serotype I-specific antibodies but not in those passively immunized with antibodies to FIPV serotype II. These data suggest that re-infection with the same serotype induces ADE in cats infected with FIPV. PMID- 19122398 TI - Adrenal diseases in ferrets in Japan. AB - Adrenal diseases are quite common in Japanese pet ferrets; however, there have been no reports concerning the epidemiology of ferret adrenal disease in Japan. The purpose of this study was to collect epidemiological data on ferret adrenal disease in Japan by sending a questionnaire to veterinarians throughout Japan. Among the 521 cases that met the criteria for this study, 307 were adrenocortical carcinoma (58.9%), 117 were adrenocortical adenoma (22.5%) and 87 were adrenocortical hyperplasia (16.7%), respectively. Sex, clinical signs, concurrent diseases, age and the sites of the affected adrenal gland in these cases were similar to those reported in North America. Most pet ferrets in Japan are imported from North America, and their husbandry in Japan is similar to that in North America, which may be the cause of the similarity in ferret adrenal disease between North America and Japan. Because a difference in the incidence of ferret adrenal diseases among countries has been reported, further research is necessary to investigate the factors related to the similarities and how to decrease the incidence of adrenal diseases in ferrets in Japan. PMID- 19122399 TI - Estrus induction and fertility rates in response to exogenous hormonal administration in postpartum anestrous and subestrus bovines and buffaloes. AB - A total of 130 animals (82 cattle, 48 buffaloes) with histories of anestrous 60 90 days post-partum and belonging to different agroclimatic zones of Punjab were subjected to rectal palpation and blood samplings at least three times at weekly intervals. The body condition score (BCS) of each animal was also recorded. The animals were divided into two groups; viz., true anestrous (Gp-I) and subestrus (Gp-II) through rectal palpation of ovaries and plasma progesterone (P4) concentrations. Furthermore, the Gp I and II animals were divided into treatment (Gp Ia, 40 cattle and 16 buffaloes; Gp IIa, 12 cattle and 14 buffaloes) and control groups (Gp Ib, 20 cattle and 8 buffaloes; Gp IIb, 10 cattle and 10 buffaloes). True anestrous animals (Gp Ia) were treated with 3 injections of hydroxyprogesterone caproate (750 mg, i.m.) at 72-hr intervals followed by injection of equine chorionic gonadotropin (eCG; 750 I.U., i.m.) 72 hr after the last progesterone injection. The animals were bred at the first estrus after the induced one. The first service conception rate (FSCR), overall conception rate (OCR), services per conception and pregnancy rate of the true anestrous treated cattle (Gp Ia) were 44.4%, 48.0%, 2.08 and 60.0%, respectively. In the true anestrous control cattle (Gp Ib), only five that were observed to be in estrus failed to conceive. In the anestrous treated buffaloes (Gp Ia), the FSCR, OCR, services per conception and pregnancy rate were 50.0%, 62.5%, 1.6 and 62.5%, respectively. No buffalo amongst true anestrous control (Gp Ib) showed estrus. The subestrus animals (Gp IIa) were administered Prostaglandin F(2alpha) (PGF(2alpha); 25 mg Dinoprost, i.m.) and bred at induced estrus. Amongst the Gp IIa animals, all cattle (100%) and twelve buffaloes (85.7%) responded to treatment. Of these animals, the FSCR and pregnancy rate at induced estrus in the cattle were 50.0% each, whereas they were 66.6% and 57.1%, respectively, in the buffaloes. The subestrus control animals (Gp IIb) remained infertile. In summary, the plasma P(4) profile can be used to differentiate true anestrous and subestrus animals and thus to determine a hormonal therapy. Furthermore, fertile estrus can be induced with hormonal therapy in anestrous and subestrus bovines. PMID- 19122400 TI - Expression of SART-1 mRNA in canine squamous cell carcinomas. AB - SART-1, a squamous cell carcinoma (SCC) antigen recognized by cytotoxic T lymphocytes, has been useful in human cancer therapy. The SART-1(259) peptide is a potential candidate for vaccine. The present study examined an orthologue of the mRNA coding this peptide in canine SCCs. Specimens were obtained from seven canine patients with SCC, and the mRNA was isolated from the samples. The SART-1 and beta-actin genes were amplified by reverse-transcription polymerase chain reaction, using the isolated mRNA as a template. Canine SART-1 was amplified in six of the seven specimens, while beta-actin was detected in all the samples. In dogs, carcinomas expressing SART-1 could be a target for cytotoxic T lymphocyte mediated immunotherapy. PMID- 19122401 TI - Juvenile diabetes mellitus accompanied by exocrine pancreatic insufficiency in a dog. AB - A 6-month-old male crossbred dog weighing 0.78 kg was presented with acute bilateral immature cataracts, intermittent diarrhea and growth retardation. The clinical manifestations and laboratory findings were suggestive of concurrent juvenile diabetes mellitus (DM) and exocrine pancreatic insufficiency (EPI). Moreover, the DM was associated with a decreased level of serum insulin-like growth factor I. Histological examination revealed a markedly lower number of pancreatic islets and acinar cells. This case shows that juvenile-onset DM can occur simultaneously with EPI and result in growth retardation, acute cataract formation and a high cortisol concentration. PMID- 19122402 TI - Existence of avian infectious bronchitis virus with a European-prevalent 4/91 genotype in Japan. AB - Eight isolates of infectious bronchitis virus (IBV) were obtained from various prefectures in Japan during 2003-2007 and were genetically analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) coupled with direct sequencing. These IBV isolates were classified into three genetic groups, including two that have already been reported (JP-I and JP-III). The remaining group is related to the 4/91 (also known as 793/B) type, prevalent mainly in European countries, and has not been identified in Japan until now. PMID- 19122404 TI - Suprascapular nerve paralysis due to streptococcal meningoradiculitis in a cow. AB - A cow, presenting with lameness with atrophy of the right supraspinatus and infraspinatus muscles, was clinically diagnosed with suprascapular nerve paralysis. Histological examination revealed necrosuppurative lesions with Gram positive cocci arranged in chains in multiple organs, including the cardiac valves, lungs, muscles, joints, brain, cerebral and spinal meninges. Spinal meningitis progressed into the roots of the right 6th to 8th cervical nerves. The suprascapular nerve showed partial loss of nerve fibers in the periphery, and innervated muscle fibers were atrophied. Bacterial culture of the cerebrospinal fluid revealed the presence of Streptococcus pyogenes. From these findings, the suprascapular nerve paralysis in this case was considered to result from meningoradiculitis associated with systemic streptococcal infection. This is a rare bovine case of suprascapular nerve paralysis due to central nerve damage of infectious cause. PMID- 19122403 TI - Salmonella prevalence in slaughtered buffaloes and pigs and antimicrobial susceptibility of isolates in Vientiane, Lao People's Democratic Republic. AB - This is the first report regarding isolation of Salmonella from cecum samples of buffaloes and pigs and characterization of the isolates in Laos. The organisms were isolated from 8% (4/50) of buffaloes and 76% (37/49) of pigs. In buffaloes, 3 animals harbored serotype 9,12: -:1,5, and 1 animal harbored both S. Derby and S. Javiana. In pigs, the most predominant serotypes were S. Derby (51%) followed by S. Anatum (45%), S. Weltevreden (15%) and S. Stanley (5%). The buffalo isolates were susceptible to the antimicrobials tested, whereas the pig isolates showed 10 resistance patterns to 1-5 antibiotics. Of the 59 pig isolates, the resistance rates to tetracycline, streptomycin, ampicillin, sulfamethoxazole trimethoprim, chloramphenicol, amoxicillin-clavulanic acid and nalidixic acid were 24%, 22%, 14%, 5%, 2%, 2% and 2%, respectively. The results suggest that pigs and buffaloes harbor Salmonella, with a higher prevalence especially in pigs, and all the isolates showed sensitivity to cefotaxime, norfloxacin and ciprofloxacin. PMID- 19122405 TI - Differentiation of foot-and-mouth disease virus-infected pigs from vaccinated pigs using a western blotting assay based on baculovirus-expressed nonstructural proteins 2C and 3D. AB - Baculovirus-expressed foot-and-mouth disease virus (FMDV) nonstructural proteins 2C and 3D were used as the antigens in a western blotting assay. This assay allowed for differentiation of FMDV-infected pigs from vaccinated pigs. Serial studies were performed using sera collected from experimentally infected and vaccinated pigs. Positive reactions were found in the sera derived from the experimentally infected pigs. There was, however, no positive reaction in the vaccinated pigs. These results suggest that this western blotting assay is a useful method for differentiation of infected pigs from vaccinated pigs. PMID- 19122406 TI - Mice are not susceptible to hepatitis E virus infection. AB - To determine whether or not mice are susceptible to hepatitis E virus (HEV) infection, C57BL/6 mice were experimentally infected with genotypes 1, 3 and 4 HEV by intravenous injection. Serum and stool samples were collected and used to detect HEV RNA and anti-HEV antibodies by RT-PCR and ELISA. The virus infection was monitored up to two months after inoculation; however, none of the serum or stool samples was positive for virus replication, demonstrating that C57BL/6 mice were not susceptible to HEV. PMID- 19122407 TI - Developmental stage- and organ-specific expression profiles of asparaginyl endopeptidases/legumains in the ixodid tick Haemaphysalis longicornis. AB - We previously identified two cDNAs from the midgut of adult Haemaphysalis longicornis that encode asparaginyl endopeptidases/legumains, HlLgm and HlLgm2. Functionally, both recombinant HlLgm and HlLgm2 efficiently digested blood proteins, haemoglobin and bovine serum albumin. Here, we investigated the expression profiles of legumain genes in the developmental stages in the life cycle of H. longicornis and in different tissues of adult ticks. Both HlLgm and HlLgm2 were well expressed in larvae, nymphs and adults. Legumain transcripts were expressed specifically in the midgut and were localized in some digestive vacuoles of gut epithelial cells. Furthermore, expression of either transcript was up-regulated by blood feeding in larvae and nymphs, suggesting the important roles of legumains in blood feeding and blood-meal digestion in ticks. PMID- 19122408 TI - Determination of fecal shedding rates and genotypes of swine hepatitis E virus (HEV) in Korea. AB - Hepatitis E virus (HEV) infection induces an acute hepatitis or a subclinical disease in humans. It is known that HEV is a zoonotic agent and pigs are major reservoirs of HEV. This study was conducted to determine the fecal shedding rates of HEV in various age groups of pigs and identify the genotypes of swine HEV prevailing in Korea. A total of 565 fecal samples were collected from suckling piglets, post-weaning pigs, growing pigs, and sows at 12 swine farms. RT-PCR was used to detect the presence of swine HEV in the feces. Every swine farm examined in this study had HEV-infected pigs. The fecal shedding rates of the swine HEV at individual farms were in the range of 2.1-35.4%. The overall fecal shedding rate of HEV in individual pigs was 17.5%. The HEV shedding rates of suckling piglets, post-weaning pigs, growing pigs and sows in their feces were 6.3, 16.3, 38.0 and 9.3%, respectively. When the genotypes of swine HEVs identified in this study were determined, they were all grouped into genotype 3. They were further subdivided into subtype 3a together with human and swine HEVs isolated in the U.S.A. PMID- 19122409 TI - Membranoproliferative glomerulonephritis in a young cat. AB - A 9-month-old male Japanese domestic cat showed pleural effusion, ascites, azotemia, hypoproteinemia and severe proteinuria. Histopathology of the percutaneous renal biopsy specimen revealed that all glomeruli showed intense mesangial hypercellularity with an increased mesangial matrix and thickening of the capillary walls, resulting in lobular accentuation of the glomerular tufts. Frequent duplication of the capillary walls was also observed. Immunostaining for alpha-smooth muscle actin distinctly revealed mesangial interposition. Diffuse global and linear deposition of C3 and IgG was observed mostly along the peripheral capillary loops. Electron microscopy confirmed frequent circumferential mesangial interposition and subendothelial dense-deposits in the glomerulus. The glomerular lesion was consistent with human membranoproliferative glomerulonephritis type I, and might be a rare case that developed at young age. PMID- 19122410 TI - Detection of Equid herpesvirus 9 DNA in the trigeminal ganglia of a Burchell's zebra from the Serengeti ecosystem. AB - Equid herpesvirus 9 (EHV-9) was isolated from a herd of Thomson's gazelles affected by encephalitis. The natural host of EHV-9 is unknown, but zebras are suspected to be the source of infection in gazelles. To prove this hypothesis, we analyzed 43 sera from Burchell's zebras (Equus burchelli) and 21 Thomson's gazelles (Gazella thomsoni) from the Serengeti ecosystem for neutralizing antibodies. Seven zebra sera were positive for EHV-1, EHV-9 and EHV-1 from Grevy's zebra strains T965 and T616. The trigeminal ganglia of 17 other Burchell's zebras and one Thomson's gazelle were tested by EHV-9 gB and EHV-1 ICP0-specific nested polymerase chain reaction (PCR). PCR sequencing confirmed that one zebra ganglion was positive for EHV-9. These results suggest that the Burchell's zebras were exposed to EHV-9 and latently infected. PMID- 19122411 TI - Focus assay on RD114 virus in QN10S cells. AB - Cats harbor an infectious endogenous retrovirus, named RD114 virus. It is therefore necessary to monitor RD114 virus production in feline cells which are used for biological products as substrates. In this study, a feline sarcoma positive leukemia-negative (S+L-) fibroblast cell line, named QN10S cells, was found to be highly susceptible to RD114 pseudotype viruses. The cells were transformed by infection with RD114 virus and the numbers of foci could be counted. The sensitivity of the focus assay was lower than that of the LacZ marker rescue assay in detecting RD114 virus. Although the assay is not suitable to detect a small amount of the virus, the assay will be useful for virological studies of RD114 virus. PMID- 19122412 TI - Enzyme-linked immunosorbent assay for screening of canine brucellosis using recombinant Cu-Zn superoxide dismutase. AB - Brucella canis, a facultative intracellular pathogen, is the causative agent of canine brucellosis. The diagnosis of canine brucellosis is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. In this study, recombinant B. canis Cu-Zn superoxide dismutase (SOD) was used as an antigen for the enzyme-linked immunosorbent assay (ELISA). The recombinant SOD showed a specific reaction with serum infected with B. canis in Western blotting and ELISA. These results suggest that ELISA using recombinant SOD is useful in screening for canine brucellosis. PMID- 19122413 TI - Measurement of brain trace elements in a dog with a portosystemic shunt: relation between hyperintensity on T1-weighted magnetic resonance images in lentiform nuclei and brain trace elements. AB - Prior to euthanasia, brain magnetic resonance imaging (MRI) was performed for a five-year-old male Yorkshire Terrier following portosystemic shunt (PSS) surgical attenuation. Hyperintensity was observed on T1W images of the lentiform nuclei. Trace elements in this area were measured by inductively coupled plasma atomic emission spectrometry. The manganese concentration in the lentiform nuclei was four times higher than that in the control group. Therefore, the manganese accumulation would be the substance that causes the hyperintensity on T1W images of the lentiform nuclei in PSS dogs. PMID- 19122414 TI - Feline digital phaeohyphomycosis due to Exophiala jeanselmei. AB - The black nodule measuring 1 cm in diameter developed in the base of nail of an 8 year-old Japanese domestic male cat. Histological examination of the excised nodule revealed a granulomatous lesion extending from the epidermis to adjacent bone. The lesion was consisted of diffuse infiltration of macrophages with epithelioid cells and multinucleated giant cells. These macrophages contained a few to numerous yeast-like brown pigmented fungus cells with a spherical shape and dark thick wall. The PCR amplification with universal primers of the 28S ribosomal RNA gene yielded a 628-bp fragment and the direct sequence confirmed that the diagnosis of the lesion was phaeohyphomycosis caused by the pathogenic dematiaceous fungus, Exophiala jeanselmei. PMID- 19122415 TI - Annual changes in serum leptin concentration in the adult female Japanese black bear (Ursus thibetanus japonicus). AB - In the present study, assay of the serum leptin concentration of the Japanese black bear (Ursus thibetanus japonicus) was attempted using a canine-leptin specific sandwich enzyme-linked immunosorbent assay (ELISA). The dose-response curve of the bear serum was linear and parallel to the canine leptin standard curve. In mated and unmated bears, the serum leptin concentration was stable at low levels from May to August or September, gradually increased from September or October, and then remarkably increased in late November. We conclude that this method may be useful for measuring bear serum leptin concentration and that the serum leptin concentration changes annually with a peak in late November. PMID- 19122416 TI - Differential immunoreactivity of microglial and astrocytic marker protein in the hippocampus of the seizure resistant and sensitive gerbils. AB - In the present study, we compared differences in ionized calcium-binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) immunoreactivities for microglia and astrocytes, respectively, in the hippocampus of the seizure resistant (SR) and seizure-sensitive (SS) gerbils. The density of Iba-1 immunoreactive microglia in the hippocampal CA1 region (CA1) and dentate gyrus (DG) of the SS gerbil was higher than that in the SR gerbil, and many Iba-1 immunoreactive microglia in the SS gerbil were hypertrophied in morphology. In contrast, we could not find significant difference in the density of GFAP immunoreactive astrocytes between the SR and SS gerbils. This result indicates that Iba-1 immunoreactive microglia in CA1 and DG of the SS gerbil are activated compared to those in the SR gerbil. PMID- 19122417 TI - [Hisayama study and gastrointestinal disease]. PMID- 19122418 TI - [Breakthrough of enteroscopic diagnoses for small intestinal diseases]. PMID- 19122419 TI - [Advances in endoscopic treatment for small intestinal diseases]. PMID- 19122421 TI - [Case of severely disabled child with refractive respiratory infection due to gastroesophageal reflux successfully controlled by using a button-shaped double lumen transgastric jejunal feeding tube]. AB - A 12-year-old severely disabled woman child had been suffering from the refractive respiratory infection due to gastroesophageal reflux (GER) in years. However two transnasal catheters inserted to control GER, one was for feeding to the jejunum and the other was for decompression of the stomach, they were not effective against respiratory infection. Then, to resolve the problems, a button shaped double lumen transgastric jejunal catheter was inserted into her jejunum via PEG in two-stage. After the procedure, the refractive respiratory infection due to GER could be successfully controlled. Additionally, by using the button shaped catheter, any position came to be acceptable in daily life, for example in rehabilitation, sleeping and so on. Her ADL (activity of daily life) was well preserved. PMID- 19122422 TI - [Bacillus cereus bacteremia in Crohn's disease with multiple ileal stricture on maintenance azathioprine therapy]. AB - We describe a case of 36-year-old Japanese man with Crohn's disease, complicated by Bacillus cereus bacteremia on maintenance azathioprine therapy. Although anti microbial agents were ineffective, the patient became well immediately after a partial resection of the ileum with multiple severe stenosis. PMID- 19122423 TI - [Case of scattered gastritis associated with ulcerative proctitis]. AB - A 59-year-old man who had ulcerative proctitis for 18 years visited our hospital because of stomach pain. Proctitis had been in remission stage for five recent years by the mesalazine administration. Esophagogastroduodenoscopy (EGD) showed scattered patchy erosions at the corpus of the stomach. Anti-acid secretory agents was administrated, however, erosive change worsened in multiplicity and in area. Biopsied specimen of gastric mucosa showed specific findings resembling to cryptitis, crypt abcess and focally enhanced gastritis. H. pylori infection was negative by some examinations. Our patient had no history of taking non steroidal anti-inflammatory drugs (NSAIDs) so far. From the findings above, it was considered that gastric lesion was strongly related to the ulcerative proctitis. 5-aminosalicylic acid (750 mg, three times daily) ground to powder was administered with predonisolon (20 mg, once daily). Five month later, all the erosions disappeared completely on EGD and biopsied specimen revealed a reduction of inflammatory cells. The present case has a rare gastric lesion with patchy pattern (not diffuse pattern) which is strongly associated with ulcerative proctitis. PMID- 19122424 TI - [Modified FOLFOX6 was effective for advanced adenocarcinoma with unknown origin in a patient with Crohn's disease]. AB - A 47-year-old man who was diagnosed as Crohn's disease at the age of 27 became aware of a mass on the forehead. CT and PET scan revealed many scattered tumors in his body. The CEA level was 4424 ng/ml. Primary lesion was not detected by the surveillance of whole gastro-intestinal tract. Liver tumor biopsy samples were histologicaly analyzed and were diagnosed as adenocarcinoma. Further immunohistochemical analysis revealed that the biopsy material had colonic character determined by positive CK20 and negative CK7 staining. Therefore, we performed 12 cycles of mFOLFOX6 chemotherapy. It was effective with the reduced size of tumors and the decreased level of CEA. PMID- 19122425 TI - [One case that accompanied accessory papilla carcinoid for von Recklinghausen's disease]. AB - A 45-year-old man: pointed out von Recklinghausen disease (following vR disease) in 18 years old. He had a checkup in a close inspection purpose of a duodenum tumor at our hospital. We diagnosis that the accessory papilla carcinoid, and Pancreas Divisum was doubted. Rocal resection of the accessory papilla was performed and picked out carcinoid of 7 mm size. In literatures searches, as for accessory papilla carcinoid, merger frequency of a papilla tumor was high in the example, and merged, vR disease of lymph node metastasis. PMID- 19122426 TI - [Case of magnetic compression anastomosis between the jejunum and the ascending colon after perforation of ileum due to intestinal tuberculosis complicated acquired immuno-deficiency syndrome]. AB - In May in 2006, a 48-year-old man admitted to our hospital for the treatment of tuberculosis and AIDS. Three weeks before the admission, he had been undergone emergency ileo-cecal resection due to the tubercular perforations of ileum and peritonitis. The double stomas of jejunum and ascending colon had made and been separated each other. When oral intake was started, a lot of intestinal juice was discharged from the stoma. Because of low level of white blood cell, the anastomosis between jejunum and ascending colon was suspended for a while. In March in 2007, magnetic compression anastomosis using Yamanouchi's method was performed for care of the dehydration and renal function due to a lot of jeunal juice discharge. With radiography, two magnets were placed in the jeunum and ascending colon through the stomas severally, and attached the walls of the intestines. Eight days after the maneuver, the anastomosis was completed without complication. Two months later, with general anestasia, the stomas were closed. PMID- 19122427 TI - [Case of laparoscopic cholecystectomy for a double gallbladder]. AB - A case of a double gallbladder, a congenital anomaly of biliary system is reported in a 71-year-old woman presenting with right hypochondrial pain. After a confirmed diagnosis of double gallbladder with gallstones was made by MRCP or ERCP, both gallbladders were removed laparoscopically. Double gallbladder is rare anatomic malformation of the biliary tract with only 88 cases, including ours, reported in the Japanese literature. Detailed preoperative investigations are required for an accurate preoperative diagnosis especially when considering laparoscopic removal to avoid inadvertent damage to biliary ductal system. PMID- 19122428 TI - [Synchronous double invasive ductal carcinomas of the pancreas with multifocal branch duct intraductal papillary mucinous neoplasms of the pancreas]. AB - A 52-year-old man with a history of distal gastrectomy for gastric cancer was admitted to our hospital because of jaundice. CT scan revealed double tumors in the pancreatic head and body concomitant with multicystic lesions of the pancreas. Total pancreatectomy with splenectomy and remnant gastrectomy was performed. Final histological diagnosis was double invasive ductal carcinomas of the pancreas head and tail with multifocal branch duct intraductal papillary mucinous adenomas of the pancreas. The present case suggests that entire pancreas might have malignant potential in patients with intraductal papillary mucinous neoplasms. PMID- 19122429 TI - [Advanced targeting therapy by antibody drugs]. PMID- 19122430 TI - [Potelligent antibodies as next generation therapeutic antibodies]. AB - Antibody-dependent cellular cytotoxicity (ADCC), a lytic attack on antibody targeted cells, is triggered upon binding of lymphocyte receptors (FcgammaRs) to the antibody constant region. ADCC is considered to be a major therapeutic function of antibodies. ADCC requires the presence of oligosaccharides in the Fc region and is sensitive to change in the oligosaccharide structure. We have demonstrated that fucose is the most critical IgG1 oligosaccharide component, and the removal of fucose from IgG1 oligosaccharides results in a very significant enhancement of ADCC and anti-tumor activity in vivo. Many therapeutic antibodies approved or clinical development are produced using Chinese hamster ovary (CHO) cells that express high level of alpha1,6-fucosyltransferase and consequently produce highly fucosylated antibodies. We have established the fucosyltransferase knockout CHO cells which could stably produce non-fucosylated antibodies, designated as Potelligent antibodies. Potelligent antibodies show potent ADCC upon target cells through the effective and antigen-specific activation of NK cells due to augmented binding to FcgammaRIIIa. Moreover, Potelligent antibodies can evade the inhibitory effect of plasma IgG on ADCC through its high FcgammaRIIIa binding. Thus, the application of Potelligent antibodies is expected to be a promising approach as next-generation therapeutic antibodies with improved efficacy, even when administered at low doses in humans in vivo. PMID- 19122431 TI - [Creation of valuable antibodies by an in vitro antibody generation system using a hypermutating B cell line]. AB - Monoclonal antibodies (mAb) have recently proven to be excellent biopharmaceutical agents. The generation of hybridomas from antigen-stimulated B cells has been a key technology for obtaining mAbs; however, it is a laborious and time-consuming process, and sometimes mAbs for molecules conserved between species are difficult to obtain because of immunological tolerance. Thus, it is of great importance to develop in vitro technologies for generating useful Abs as drug candidates. We have been attempting to develop a novel in vitro antibody generation system using a chicken B cell line DT40, which displays Abs and mutates Ig genes during culture, thereby generating a useful Ab library for screening mAbs. First, we generated an engineered cell line DT40-SW whose mutation machinery can be reversibly switched on and off. The Ab generation system using DT40-SW is useful in the following ways: (1) mAbs for various model antigens including antoantigens can be obtained from the DT40-SW Ab library that is free from immunological tolerance; (2) the switching device of the mutation machinery enables fixing desirable Ig mutants by stopping mutation; (3) by repeated culture and sorting of clones bearing higher affinity for target antigens, affinity maturation can be mimicked in vitro. We have also genetically improved DT40-SW cells for mutation efficiency and Ab production. The Ab generation system will be applicable for obtaining valuable Abs such as antitumor Abs. PMID- 19122432 TI - [Targeting therapy for inflammatory diseases by anti-TNFalpha biologics]. AB - TNFalpha (tumor necrosis factor-alpha) plays a critical role in the pathogenesis of inflammatory diseases including rheumatoid arthritis and Crohn's disease. Infliximab is a monoclonal antibody that recognizes human TNFalpha. Clinical trials have been persuasive that infliximab is effective and far superior to the conventional drug therapy in various inflammatory diseases. Combination of infliximab plus methotrexate is effective in patients with active rheumatoid arthritis who have not responded adequately to traditional disease-modifying anti rheumatic drugs, and has produced significant improvement in clinical, radiographic, and functional outcomes. Infliximab is also an important treatment option in patients with active Crohn's disease who have not responded to conventional therapy and in those with this disease who have fistulae. Moreover, infliximab treatment has resulted in effective suppression of ankylosing spondylitis, psoriasis and ocular inflammation in patients with refractory uveoretinitis due to Behcet's disease. Thus, biologics targeting TNFalpha have revolutionized the therapy of inflammatory diseases. Here, the current status of clinical application of anti-TNFalpha biologics is reviewed by describing the clinical outcome of infliximab and future prospects of biologics are discussed. PMID- 19122433 TI - [Therapeutic effect of anti-nucleokine monoclonal antibody on ischemic brain infarction]. AB - Ischemic brain infarction is high among th causes of death in Japan, and the medical and social burden by severe sequela is also extremely serious. In this symposium, we show that treatment with anti-high mobility group box 1 (HMGB1) monoclonal antibody (mAb) remarkably ameliorated brain infarction induced by 2 hour occlusion of the middle cerebral artery in rats, even when the mAb was administered after the start of reperfusion. Whereas HMGB1 is usually localized in nucleus, after stimulation it is secreted into extracellular space by an unknown non-classical pathway, and exhibits an inflammatory cytokine-like activity. Treatment with mAb reduced infarct size, and the accompanying neurological deficits in locomotor function were significantly improved. In addition, some biochemical markers such as permeability of the blood-brain barrier, the expression of tumor necrosis factor-alfa, inducible nitric oxide synthase and matrix metalloproteinase-9 were altered by mAb injection. These findings indicate the usefulness of HMGB1 as a novel therapeutic to target ischemic stroke. PMID- 19122434 TI - [Development of technology for safely decontaminating and decomposing chemical and biological agents]. PMID- 19122435 TI - [Decomposition and treatment of environmental pollutants]. AB - The wastes, which have explosiveness, toxicity, infectious, and possibility to cause damage to other people's health and environments, are provided the special management municipal wastes or the special management industry wastes. All the treatment processes from collection to disposal is done under strict management. In this report, we focus on the inorganic and organic experimental waste fluid and the infectious waste among the special management municipal waste or industry waste, because of the similar toxic and chemical characteristics to that of biological and chemical weapons. Understanding how the decomposition and treatment of these wastes are actually done, we clarify the issues of the hazardous waste management and discuss on the possibility to develop the new degradation and treatment method for biological and chemical weapons. PMID- 19122436 TI - [Chemical treatment and decomposition technique of the chemical warfare agents containing arsenicals]. AB - The old Japanese army developed several chemical warfare agents on Ohkuno Island in Seto inland sea, Hiroshima Japan, during the period between 1919 and 1944. These chemical agents including yperite (mustard; irritating agent), lewisite (irritating agent), diphenylchloroarsine (DA; vomiting agent), diphenylcyanoarsine (DC; vomiting agent) and other poisonous gases were manufactured to be used in China. After World War II, the old Japanese army abandoned or dumped these agents into seas inside or outside of Japan and interior of China. Rather than being used for terrorism, these chemical warfare agents containing arsenicals may cause injury to some workers at the digging site of abandoned chemical weapons. Moreover, the leakage of chemical agents or an explosion of the bomb may result in environmental pollution, as a result, it is highly possible to cause serious health damage to the residents. There are still many abandoned or dumped warfare agents in Japan and China, therefore chemical agents containing arsenic are needed to be treated with alkaline for decomposition or to decompose with oxidizing agent. Presently, a large quantity of chemical agents and the contaminated soil are processed by combustion, and industrial waste is treated with sulfur compounds as the insoluble sulfur arsenic complex. This report describes the methods for the disposal of these organic arsenic agents that have been implemented until present and examines the future prospects. PMID- 19122437 TI - [Decontamination of chemical and biological warfare agents]. AB - Chemical and biological warfare agents (CBWA's) are diverse in nature; volatile acute low-molecular-weight toxic compounds, chemical warfare agents (CWA's, gaseous choking and blood agents, volatile nerve gases and blister agents, nonvolatile vomit agents and lacrymators), biological toxins (nonvolatile low molecular-weight toxins, proteinous toxins) and microbes (bacteria, viruses, rickettsiae). In the consequence management against chemical and biological terrorism, speedy decontamination of victims, facilities and equipment is required for the minimization of the damage. In the present situation, washing victims and contaminated materials with large volumes of water is the basic way, and additionally hypochlorite salt solution is used for decomposition of CWA's. However, it still remains unsolved how to dispose large volumes of waste water, and the decontamination reagents have serious limitation of high toxicity, despoiling nature against the environments, long finishing time and non durability in effective decontamination. Namely, the existing decontamination system is not effective, nonspecifically affecting the surrounding non-target materials. Therefore, it is the urgent matter to build up the usable decontamination system surpassing the present technologies. The symposiast presents the on-going joint project of research and development of the novel decontamination system against CBWA's, in the purpose of realizing nontoxic, fast, specific, effective and economical terrorism on-site decontamination. The projects consists of (1) establishment of the decontamination evaluation methods and verification of the existing technologies and adaptation of bacterial organophosphorus hydrolase, (2) development of adsorptive elimination technologies using molecular recognition tools, and (4) development of deactivation technologies using photocatalysis. PMID- 19122438 TI - [Decontamination of chemical warfare agents by photocatalysis]. AB - Photocatalysis has been widely applied to solar-energy conversion and environmental purification. Photocatalyst, typically titanium dioxide (TiO(2)), produces active oxygen species under irradiation of ultraviolet light, and can decompose not only conventional pollutants but also different types of hazardous substances at mild conditions. We have recently started the study of photocatalytic decontamination of chemical warfare agents (CWAs) under collaboration with the National Research Institute of Police Science. This article reviews environmental applications of semiconductor photocatalysis, decontamination methods for CWAs, and previous photocatalytic studies applied to CWA degradation, together with some of our results obtained with CWAs and their simulant compounds. The data indicate that photocatalysis, which may not always give a striking power, certainly helps detoxification of such hazardous compounds. Unfortunately, there are not enough data obtained with real CWAs due to the difficulty in handling. We will add more scientific data using CWAs in the near future to develop useful decontamination systems that can reduce the damage caused by possible terrorism. PMID- 19122439 TI - [Highly sensitive detection technology for biological toxins applying sugar epitopes]. AB - The Shiga toxin is a highly poisonous protein produced by enterohemorrhagic Escherichia coli O157. This bacterial toxin causes the hemolytic uremic syndrome. Another plant toxin from castor beans, ricin, is also highly toxic. The toxin was used for assassination in London. Recently, there were several cases of postal matter containing ricin. Both toxins are categorized as biological warfare agents by the Centers of Disease Control and Prevention. Conventional detection methods based on the antigen-antibody reaction, PCR and other cell-free assays have been proposed. However, those approaches have drawbacks in terms of sensitivity, analytical time, or stability of the detection reagents. Therefore, development of a facile and sensitive detection method is essential. Here we describe new detection methods applying carbohydrate epitopes as the toxin ligands, which is based on the fact that the toxins bind cell-surface oligosaccharides. Namely, the Shiga toxin has an affinity for globobiosyl (Gb(2)) disaccharide, and ricin binds the beta-D-galactose residue. For Shiga toxin detection, surface plasmon resonance (SPR) was applied. A polyanionic Gb(2)-glycopolymer was designed for this purpose, and it was used for the assembly of Gb(2)-chips using alternating layer-by-layer technology. The method allowed us to detect the toxin at a low concentration of LD(50). A synthetic carbohydrate ligand for ricin was designed and immobilized on the chips. SPR analysis with the chips allows us to detect ricin in a highly sensitive and facile manner (10 pg/ml, 5 min). Our present approaches provide a highly effective way to counter bioterrorism. PMID- 19122440 TI - [Overview of 40 years' chemical study]. AB - 1,6-Dihydro-3(2H)-pyridinone, designed as a common synthon for synthesis of various natural products, was found to be easily prepared in large scale and successfully used to synthesize a variety of alkaloids such as ibogamine, quinine and tecomanine. A tricyclo[3.3.0.0(2.8)]octane was also served as a common synthon for several sesquiterpenes such as pentalenene and quadrone. Synthetic studies by using sulfinyl chirality via an intramolecular Michael addition gave the novel route to construct spiro-ketal moiety in enantiomerically pure form. By applying this method, many natural spiro-ketal compounds were asymmetrically synthesized effectively. 3-Sulfinylated 1,4-dihydropyridine, a chiral NADH model compound, reduced activated ketones such as methyl benzoylformate to give the corresponding alcohols in excellent optical yields. A kind of 3-O-substituted pyridoxal chiral model compound was useful for preparation of alpha,alpha dialkylated alpha-amino acids by asymmetric alpha-alkylation of alpha-amino acids. 2'-O,4'-C-Bridged nucleic acid analogs, BNAs, developed as novel type of artificial nucleic acids, showed an extraordinarily high binding affinity toward single stranded RNA and double stranded DNA complements along with excellent nuclease-resistant ability. Oligonucleotides containing BNA monomer units were proved to be very useful for various biotechnologies, such as antisense and antigene methodologies. PMID- 19122441 TI - [Perspectives on the current state of pharmacogenomics in drug development]. AB - Pharmacogenomics is expected to become one of the ways by which serious drug development problems can be broken down and solved. In fact, the field of pharmaceutical development seems to be using pharmacogenomics increasingly as a means of both drug selection (via genotyping) and proper dosage determination. Before pharmacogenomics can be put to practical use, however, scientific and technical issues must first be resolved, after which social and ethical issues must be addressed. In Japan, drug developers are preparing for the introduction of pharmacogenomics into clinical trials. As they anticipate the necessary revisions, they must keep in mind not only the differences between current practice and that including pharmacogenomics, but also international standards. Therefore, developers are discussing strategies for communicating the necessary changes to academic and regulatory parties in an attempt to obtain a consensus and smoothly implement these changes. A survey of the academic and regulatory parties revealed that there were concerns about what pharmacogenomic information should be obtained, who would have access to it, and how it should be transmitted. Since industry, academia, and the regulatory body all agree that pharmacogenomics need to be implemented in Japan, deeper discussion of the science, technology, regulation, and ethics relevant to this topic should be continued both domestically and internationally. PMID- 19122442 TI - [methylglyoxal-induced superoxide anion production in endothelial cells]. AB - Methylglyoxal (MG), a highly reactive dicarbonyl compound, is a metabolic by product of glycolysis. MG is often detected at high levels in the blood of diabetic patients. We examined whether MG was capable of inducing reactive oxygen species (ROS) production in bovine aortic endothelial cells (BAECs). The viability of BAECs decreased with time on treatment with 5 mM MG, and was almost completely lost at 24 h. In contrast, MG at 1 mM had little influence on BAEC viability up to 24 h, but induced the elevation of intracellular glutathione content at 24 h. Exposure of BAECs to MG caused a dose-dependent increase in oxidized-hydroethidine fluorescence intensity, indicating ROS production. In addition, aconitase inactivation, which is an indicator of intracellular superoxide, was observed in MG-treated cells. Finally, we found that MG at 5 mM increased the fluorescence intensity of BES-So, a specific probe for superoxide. Together, the results suggest that MG induces superoxide production in endothelial cells, and that the accumulation of ROS may be linked to cytotoxic effects. PMID- 19122443 TI - [Effect of the brand and generic medicine of pravastatin on dyslipidemia in rabbits fed a high cholesterol diet]. AB - Mevalotin containing pravastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, is the brand medicine and well known to be effective for patients with dyslipidemia. Now, more than 20 generic pravastatins are available for clinical therapy. We compared pharmaceutical property of Mevan,a generic pravastatin, with that of Mevalotin.According to the definition of the Japanese Pharmacopoeia, Mevalotin 10 mg tablets were uniform in pravastatin content, whereas 5 mg tablets were rather variable. Variation in pravastatin content of Mevan 5 mg tablets was the same as Mevalotin 5 mg, whereas that of 10 mg tablets was very variable. The plasma concentration of pravastatin in the normal rabbits continuously increased until 180 min after oral administration of 30 mg Mevan, whereas it increased in a biphasic pattern after 30 mg Mevalotin.All rabbits were fed 0.2% cholesterol diet throughout the experiment. After 8 weeks, oral administration of either Mevalotin or Mevan was started at the dose of 30 mg pravastatin/day for 16 weeks. After a transient increase for a few weeks, the plasma levels of total- and LDL-cholesterol gradually decreased in Mevalotingroup, whereas these levels did not significantly changed in Mevan group within 16 weeks. The level of HDL-cholesterol in Mevan group tended to increase but not in Mevalotin group. The triglyceride level in Mevan group changed as well as that in Mevalotin group until 10 weeks after administration, and then gradually increased. The present results suggest that pharmaceutical properties of Mevan are not always identical with those of Mevalotin. PMID- 19122444 TI - [Systematic analysis of oral hypoglycemic agents in health foods]. AB - A systematic analysis for 11 ingredients of oral hypoglycemic agent in health foods was established using three different analytical methods; i.e. thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and comparison of MS/MS spectra analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). In Normal-phase and reversed-phase TLC, each condition to separate and detect 10 ingredients except nateglinide was developed. On the other hand, 11 ingredients were detected qualitatively and quantitatively by HPLC. The recovery rates were 92-101% and each coefficient of variation was less than 5.4%. Then UV spectra were monitored using this HPLC method and furthermore MS/MS spectra of 11 ingredients were obtained by LC/MS/MS. Identification of each ingredient became precise and rapid by comparing UV and MS/MS spectra of standard solutions with that of extract solutions from health foods. Using this systematic analysis, glibenclamide was accurately determined and identified from health foods. PMID- 19122445 TI - Determination of pyrethroid pesticides in cinnamomi cortex. AB - In Japan, maximum residue levels (MRL) have been set for eight pesticides (alpha BHC, beta-BHC, gamma-BHC, delta-BHC (BHCs), p,p'-DDE, o,p'-DDT, p,p'-DDD, p,p' DDT (DDTs)) in 14 crude drugs (below 0.2 ppm as total of BHCs, below 0.2 ppm as total of DDTs). There are fears that pesticides present in crude drugs for which MRL are set will be changed from BHCs and DDTs to other pesticides with MRL setting as the turning point. There are few surveys of pyrethroid pesticide in crude drugs distributed in Japan. The actual situation of pyrethroid pesticides in crude drugs distributed in Japan after setting MRL is not unclear and should be clarified. Although a method to analyze permethrin, cypermethrin and fenvalerate in 11 crude drugs was reported, it is not adequate because the recovery rates of permethrin, cypermethrin and fenvalerate from Cinnamomi cortex were very low and the method, including liquid-liquid partition is difficult. In this study, we developed a method using solid-phase extraction to analyze permethrin, cypermethrin and fenvalerate in Cinnamomi cortex with acceptable recovery rates. The sample solution was determined by gas chromatography/mass spectrometry with negative chemical ionization. The recovery rates of permethrin, cypermethrin and fenvalerate from Cinnamomi cortex were between 87.9 and 90.7%. Five samples of Cinnamomi cortex were analyzed according to the proposed method. No samples contained permethrin, cypermethrin and fenvalerate over detection limits. The proposed method could analyze permethrin, cypermethrin and fenvalerate in all crude drugs for which MRL are set. PMID- 19122446 TI - [Effort to improve advanced problem-based learning tutorial]. AB - To prepare for the introduction of the advanced problem-based learning (PBL) tutorial for higher-grade students under the six-year pharmacy curriculum, a trial of the tutorial was performed in a fourth-grade class under the former four year curriculum in 2007. A questionnaire survey conducted to identify any problems in performing the tutorial revealed: 1) the number of students in each group was too large; 2) the contents of presentations seemed to overlap due to the limited number of task cases, which forced more than one group to address a particular case; and 3) the time-line from the day of product presentation to that of periodic examination was too short to hold a sufficient group discussion. In 2008, to resolve these problems: 1) the number of groups was increased to reduce the number of students in each group; 2) new task cases were added to decrease the number of groups addressing a particular case; and 3) an adequate time period was arranged between the days of product presentation and periodic examination. The survey conducted this year demonstrated that students' learning environment had been improved by these changes in the method of performing the tutorial, but also revealed new problems such as prolongation of the time required for product presentation and the difficulty levels of task cases. In addition, the usefulness of customer satisfaction (CS) analysis was demonstrated as a result of applying it to the data analysis of evaluation performed on group presentations. PMID- 19122447 TI - Stress-induced hypertension and increased sympathetic activity in mice overexpressing neuropeptide Y in noradrenergic neurons. AB - BACKGROUND AND AIMS: Neuropeptide Y (NPY) is a sympathetic neurotransmitter co stored and co-released with noradrenaline and adrenaline. We have constructed a novel NPY transgenic mouse model (OE-NPY(DBH) mouse) where targeted overexpression results in increased levels of NPY in the brainstem and adrenal glands. The present study was aimed to understand the role of NPY released from sympathetic nerves and brain noradrenergic neurons in regulation of blood pressure, and behavioral responses to stress. METHODS: Blood pressure was measured by radiotelemetry in conscious male OE-NPY(DBH) and wild-type mice during surgical stress and in baseline conditions. Plasma and adrenal gland catecholamine levels were measured at baseline. Acute immobilization and cold exposure were used to study the plasma levels of NPY and corticosterone in stress, and brown adipose tissue thermogenic activity was measured with [(3)H]GDP binding after cold. RESULTS: Here, we demonstrate that sympathoadrenal activity is enhanced in the OE-NPY(DBH) mice. Blood pressure during surgical stress was significantly increased in comparison with wild-type controls. Furthermore, OE NPY(DBH) mice showed sexually dimorphic NPY responses to stress, and an anxiolytic-like behavior in elevated plus-maze and light-dark tests. CONCLUSION: This study shows that the overactive noradrenergic NPY system plays a role in regulation of blood pressure and adaptive responses to stress, and may be a link between chronic stress and adiposity-associated disturbances in metabolism. PMID- 19122448 TI - Central ghrelin affects pituitary-thyroid axis: histomorphological and hormonal study in rats. AB - Body weight depends on the balance between energy intake and consumption. An interaction between ghrelin and thyroid function has been reported only in pathophysiological states. We examined whether intracerebroventricular (ICV) administration of ghrelin affects the structure and function of the pituitary thyroid axis in young adult male rats. Ghrelin (0.3 nmol/5 microl PBS) or an equal volume of PBS were injected every 24 h into the lateral cerebral ventricle for 5 days. Two hours after the last treatment the animals were killed, their pituitaries and thyroids excised and prepared for further histological, immunohistochemical and morphometric investigation. Serum TSH levels were measured by RIA, while the total T(4) and T(3) levels were examined by ECLIA. Ghrelin treatment increased pituitary weight (p < 0.05) when compared to the controls, with no effect on the thyroid weight. Smaller, degranulated TSH immunopositive cells were noticed within the pituitaries of ghrelin-treated animals; their cellular and nuclear volume as well as the relative volume density of thyrotrophs decreased (p < 0.05) in comparison to the control values. The level of serum TSH was reduced (p < 0.05). In the thyroid parenchyma of ghrelin treated rats, an increased number of hypofunctioning follicles was noticed, characterized by flattened, weakly Tg-immunoreactive epithelium and colloid distension. The relative volume densities of the follicles and colloid increased (p < 0.05), while the thyroid index of activation rate and the serum level of total T(4) decreased (p < 0.05). In conclusion, centrally applied ghrelin modulated the immunohistomorphometric features of pituitary TSH cells and decreased the level of serum TSH, consequently changing thyroid morphology and function, by reducing the T(4) hormone level in the serum. PMID- 19122449 TI - Flexible bronchoscope damage and repair costs in a bronchoscopy teaching unit. AB - BACKGROUND: A significant part of the budget of our bronchoscopy unit represents repair costs for damaged bronchoscopes. OBJECTIVES: The purpose of this study was to determine the frequency, type and cause of damage to the bronchoscope as well as the repair costs. METHODS: Frequency, type and cause of bronchoscope damage and repair costs of 13 new bronchoscopes that were used between August 1, 2001, and December 31, 2006, were retrospectively studied. RESULTS: We recorded 47 instances of bronchoscope damage during the study, which is 1 instance of damage/141.6 procedures. Six instances of damage (12.7%) were potentially preventable. The most frequent wear and tear damage was to the rubber sheath on the distal bending portion of flexible bronchoscopes, and the most frequently preventable damage was that of the suction channel of the bronchoscope. The repair costs totaled 34,950.00 EUR or 5.25 EUR/procedure. 17,781.00 EUR (50.9%) can be attributed to preventable damage. The use of bronchoscopes for educational purposes was not associated with a higher rate of bronchoscope damage at our institution. CONCLUSIONS: Only a small number of occurrences of bronchoscope damage in our unit are potentially preventable, but they still represent an important expense. The relatively low occurrence of preventable damage is a result of the successful bronchoscopy training program. PMID- 19122450 TI - Clonality of basal cell carcinoma arising in an epidermal nevus. New insights provided by molecular analysis. AB - Non-organoid non-epidermolytic (keratinocytic) epidermal nevi are benign congenital skin lesions following the lines of Blaschko. The occurrence of epithelial tumors in epidermal nevi of this type is very rare, and it remains elusive whether these lesions are clonally related or represent a simple collision of independent tumors. We report a case of a patient who developed 2 basal cell carcinomas in an extensive epidermal nevus on the trunk. Molecular analysis of a biopsy containing both basal cell carcinoma and epidermal nevus revealed a PIK3CA mutation. Further analysis suggested that this mutation is present in the basal cell carcinoma rather than in the epidermal nevus. The results favour an independent origin of the basal cell carcinoma, although a possible clonal relationship between both lesions cannot be completely excluded on the basis of the molecular genetic findings. PMID- 19122451 TI - C-Flex technology: effects on breathing parameters and inspiratory flow limitation. AB - BACKGROUND: Expiratory pressure relief continuous positive airway pressure (pressure relief CPAP, C-Flex) is known to be as effective in the treatment of obstructive sleep apnea (OSA) as conventional CPAP while improving overall patients' adherence. However, the effects of C-Flex on ventilation during sleep have not been studied yet. OBJECTIVE: This study investigates the effects of pressure relief CPAP on respiratory parameters and possible inspiratory flow limitation with increased difference between inspiratory and expiratory pressure compared with conventional CPAP. METHODS: In total, 24 patients were investigated both during conventional CPAP and during three C-Flex pressure relief settings in randomized order during rapid-eye-movement (REM) and non-REM (NREM) sleep. Airflow was monitored with a pneumotachograph; inspiratory flow limitation was assessed by analyzing airflow and esophageal pressure swings. RESULTS: Using higher C-Flex gains, expiratory time decreased in favor of the inspiratory duty cycle while there was no significant change in tidal volume. Analysis of inspiratory flow limitation showed no significant difference between conventional CPAP and the C-Flex gains studied. CONCLUSIONS: The increase in the inspiratory duty cycle with C-Flex might either indicate an increase in the work of breathing or a decrease in the work of breathing due to a lower peak end-expiratory pressure and consecutive alleviation of passive expiration. Both treatments appeared equivalent regarding the occurrence of inspiratory flow limitation. PMID- 19122452 TI - An analytical model for elucidating tendon tissue structure and biomechanical function from in vivo cellular confocal microscopy images. AB - Fibered confocal laser scanning microscopes have given us the ability to image fluorescently labeled biological structures in vivo and at exceptionally high spatial resolutions. By coupling this powerful imaging modality with classic optical elastography methods, we have developed novel techniques that allow us to assess functional mechanical integrity of soft biological tissues by measuring the movements of cells in response to externally applied mechanical loads. Using these methods we can identify minute structural defects, monitor the progression of certain skeletal tissue disease states, and track subsequent healing following therapeutic intervention in the living animal. Development of these methods using a murine Achilles tendon model has revealed that the hierarchical and composite anatomical structure of the tendon presents various technical challenges that can confound a mechanical analysis of local material properties. Specifically, interfascicle gliding can yield complex cellular motions that must be interpreted within the context of an appropriate anatomical model. In this study, we explore the various classes of cellular images that may result from fibered confocal microscopy of the murine Achilles tendon, and introduce a simple two-fascicle model to interpret the images in terms of mechanical strains within the fascicles, as well as the relative gliding between fascicles. PMID- 19122455 TI - Cell therapy of acute myocardial infarction: open questions. AB - Cell-based therapy is a promising option for treatment of cardiovascular diseases. Based on experimental studies demonstrating that bone marrow-derived stem and progenitor cells improve the functional recovery after ischemia, clinical trials were initiated to address this new therapeutic concept. So far, clinical studies preferentially used adult bone marrow-derived cells for the treatment of patients with acute myocardial infarction. Overall, cell therapy was shown to reduce the infarct size and improve cardiac contractile function. However, the extent of functional improvement varied between the studies and various questions remain. The present review article will discuss the heterogeneity of the clinical results, the timing of the cell delivery and the benefit achieved so far in different patient populations. PMID- 19122453 TI - Subcortical laterality of speech encoding. AB - It is well established that in the majority of the population language processing is lateralized to the left hemisphere. Evidence suggests that lateralization is also present in the brainstem. In the current study, the syllable /da/ was presented monaurally to the right and left ears and electrophysiological responses from the brainstem were recorded in adults with symmetrical interaural click-evoked responses. Responses to the right-ear presentation occurred earlier than those to left-ear presentation in two peaks of the frequency following response (FFR) and approached significance for the third peak of the FFR and the offset peak. Interestingly, there were no differences in interpeak latencies indicating the response to right-ear presentation simply occurred earlier over this region. Analyses also showed more robust frequency encoding when stimuli were presented to the right ear than the left ear. The effect was found for the harmonics of the fundamental that correspond to the first formant of the stimulus, but was not seen in the fundamental frequency range. The results suggest that left lateralization of processing acoustic elements important for discriminating speech extends to the auditory brainstem and that these effects are speech specific. PMID- 19122456 TI - Long-term partial remission of autoimmune thrombocytopenia in a patient treated with the anti-tumor necrosis factor-alpha antibody infliximab for refractory fistulizing Crohn's disease. AB - BACKGROUND: Up to the present date, the treatment of recurrent chronic fistulas occurring with Crohn's disease represents a challenging task for both internists and surgeons alike. METHODS: Conservative methods of treatment using steroids, dietotherapy, antibiotics or immunosuppressive agents are not particularly effective in treating fistulas. Treatment with anti-tumor necrosis factor-alpha (TNFalpha) antibodies results in more remissions of fistulas. However, its use requires appropriate medical experience and causes substantial costs. Surgical procedures such as fistula sanitation or protective ileostomy are often obfuscated by the recurrence of the condition. The efficiency of anti-TNFalpha antibodies for the treatment of active Crohn's disease has been evidenced through several multicenter, double-blind, placebo-controlled studies. RESULTS: Here we report the successful anti-TNFalpha treatment of a patient suffering from chronic, fistulizing, therapy-resistant Crohn's disease and a concomitant chronic autoimmune thrombocytopenia. The chimeric anti-TNFalpha antibody infliximab markedly reduced the disease activity of Crohn's disease, and, in addition, substantially increased the platelet counts. After infliximab application, no other treatment of autoimmune thrombocytopenia was required. After infliximab therapy, autoantibodies to dsDNS could be observed. However, there were no signs of a lupus-like disease. CONCLUSION: This report indicates a favorable therapeutic response of autoimmune thrombocytopenia associated to Crohn's disease. PMID- 19122457 TI - The clinical significance of serum procalcitonin levels following direct hemoperfusion with polymyxin B-immobilized fiber column in septic patients with colorectal perforation. AB - BACKGROUND: The efficacy of direct hemoperfusion with polymyxin B-immobilized fiber columns (PMX) has already been demonstrated in clinical studies for the treatment of septic shock. However, serum procalcitonin levels following PMX remain unknown. METHODS: This prospective, multicenter, nonrandomized clinical study was performed at 12 institutions. Forty-five patients with severe sepsis or septic shock due to colorectal perforation underwent PMX. Patients' outcome as well as circulating levels of endotoxin, procalcitonin and IL-6 were monitored. RESULTS: Before surgery, procalcitonin level, but not endotoxin and IL-6 levels, was elevated according to patients' septic conditions. Procalcitonin was significantly and positively correlated with sequential organ failure assessment score. Circulating levels of procalcitonin peaked 24 h after PMX treatment. Change in serum procalcitonin level was significantly higher in nonsurvivors than survivors. Nine mortalities were observed within 28 days. The best predictor for 28-day mortality was procalcitonin >85.7 ng/ml at 24 h after PMX (area under the receiver operating characteristic curve: 0.808 +/- 0.105). CONCLUSIONS: Procalcitonin may be a good indicator of severity of sepsis secondary to colorectal perforation. Furthermore, procalcitonin level at 24 h after PMX appears to predict outcome after PMX. Therefore, procalcitonin may be a useful diagnostic marker to evaluate patients' condition in candidates for PMX treatment. PMID- 19122458 TI - Degradation of collagen by metalloproteinase 2 in patients with abdominal hernias. AB - BACKGROUND: The groin hernia is a significant social and economic problem of our times. The pathogenesis of the disease is not clear. The metalloproteinases (MMP) are the group of proteolytic enzymes responsible for the degradation of extracellular matrix proteins and the basic membrane of blood vessels. THE AIMS OF OUR STUDY WERE: (1) to estimate the MMP-2 levels in the blood and tissues of patients with a groin hernia; (2) to answer the question of whether changes in MMP-2 activity correlate with the occurrence of inguinal hernias. METHOD: The study was performed on a group of 90 male patients suffering from inguinal hernias, aged 28-70 years (mean: 49 years). The control group was made up of 10 healthy (free from hernia) males, aged 30-68 years (mean: 46 years). RESULTS: We noticed increased levels of MMP-2 in patients with all types of hernia and across all age groups. The MMP-2 mean serum levels were statistically higher in patients with a groin hernia when compared to the control group. The highest blood levels of MMP-2 were observed in young men with a direct hernia. CONCLUSIONS: This study confirmed the important role of MMP-2 in the pathogenesis of inguinal hernia. The increased activity may lead to dysfunctions in collagen fiber, which is responsible for forming fascial structures, and as a result weaken their durability. PMID- 19122459 TI - Sacrococcygeal teratomas: prenatal surveillance, growth and pregnancy outcome. AB - OBJECTIVE: Prenatal surveillance and growth characteristics are evaluated in a cohort of fetuses with sacrococcygeal teratomas (SCT) as part of risk assessment. METHODS: Retrospective review of 23 fetuses with SCT: prenatal diagnosis, surveillance, delivery, and early postnatal outcome are reported. RESULTS: Cardiac output failure physiology requires serial evaluation. The size of the SCT determines obstetrical risks and mode of delivery. An SCT growth rate approaching >150 cm(3) per week may be associated with increased perinatal mortality risks. Maternal morbidity is related mainly to polyhydramnios and preterm labor. CONCLUSIONS: Perinatal mortality is approximately 43%. Maternal-fetal surgery for fetal physiologic deterioration is not frequent, but serial surveillance is required to minimize fetal morbidity/mortality and maternal morbidity. Rapid SCT growth rates may be associated with increased risk of perinatal mortality. PMID- 19122460 TI - Application of automatic speech recognition to quantitative assessment of tracheoesophageal speech with different signal quality. AB - OBJECTIVE: Tracheoesophageal voice is state-of-the-art in voice rehabilitation after laryngectomy. Intelligibility on a telephone is an important evaluation criterion as it is a crucial part of social life. An objective measure of intelligibility when talking on a telephone is desirable in the field of postlaryngectomy speech therapy and its evaluation. PATIENTS AND METHODS: Based upon successful earlier studies with broadband speech, an automatic speech recognition (ASR) system was applied to 41 recordings of postlaryngectomy patients. Recordings were available in different signal qualities; quality was the crucial criterion for this study. RESULTS: Compared to the intelligibility rating of 5 human experts, the ASR system had a correlation coefficient of r = 0.87 and Krippendorff's alpha of 0.65 when broadband speech was processed. The rater group alone achieved alpha = 0.66. With the test recordings in telephone quality, the system reached r = -0.79 and alpha = 0.67. CONCLUSION: For medical purposes, a comprehensive diagnostic approach to (substitute) voice has to cover both subjective and objective tests. An automatic recognition system such as the one proposed in this study can be used for objective intelligibility rating with results comparable to those of human experts. This holds for broadband speech as well as for automatic evaluation via telephone. PMID- 19122461 TI - Jobs, sex, love and lifestyle: when nonstutterers assume the roles of stutterers. AB - PURPOSE: This study assessed the impact of stuttering via a questionnaire in which fluent individuals were asked to assume the mindset of persons who stutter (PWS) in various life aspects, including vocation, romance, daily activities, friends/social life, family and general lifestyle. The perceived impact of stuttering through the mind's eyes of nonstutterers is supposed to reflect respondents' abilities to impart 'theory of mind' in addressing social penalties related to stuttering. METHOD: Ninety-one university students answered a questionnaire containing 56 statements on a 7-point Likert scale. Forty-four participants (mean age = 20.4, SD = 4.4) were randomly selected to assume a stuttering identity and 47 respondents (mean age = 20.5, SD = 3.1) to assume their normally fluent identity. RESULTS: Significant differences between groups were found in more than two thirds of items regarding employment, romance, and daily activities, and in fewer than half of items regarding family, friend/social life, and general life style (p <0.001). CONCLUSIONS: The social penalties associated with stuttering appear to be apparent to fluent individuals, especially in areas of vocation, romance, and daily activities, suggesting that nonstuttering individuals, when assuming the role of PWS, are capable of at least temporarily feeling the negative impact of stuttering. PMID- 19122462 TI - Anorectal function testing and anal endosonography in the diagnostic work-up of patients with primary pelvic organ prolapse. AB - AIM: To study the pathophysiology of defecation disorders in patients with primary pelvic organ prolapse (POP) and the diagnostic potential of anorectal function testing (AFT) including endosonography in the work-up of these patients. METHODS: 59 Patients were evaluated with a validated questionnaire, clinical examination, AFT and endosonography. RESULTS: Women with POP showed lower squeezing pressure, postponed first sensation and desire, lower capacity and prolonged pudendal nerve terminal latency time compared to healthy controls (all p < 0.01). Manometric findings did not differ significantly between patients with and without constipation. Patients with fecal incontinence had significantly lower resting and squeezing pressures than patients without fecal incontinence and an increased risk of an external sphincter defect (odds ratio = 12.75, 95% confidence interval 2.40-66.67). Although digital rectal examination could quantify absent, decreased and normal squeezing pressure, the positive predictive value for external sphincter defects was low (0.32). CONCLUSION: AFT indicates the presence of neuromuscular damage of the anorectal region in patients with POP. AFT is not useful in the work-up of patients with POP and constipation, because it fails to discriminate between symptomatic and asymptomatic patients. In cases of fecal incontinence, AFT and endosonography are helpful to distinguish between functional and anatomical problems. PMID- 19122463 TI - Alterations in RCAS1 serum concentration levels during menstrual cycle in patients with uterine leiomyoma and lack of analogical changes in adenomyosis. AB - INTRODUCTION: The selective suppression phenomenon of the cytotoxic immune response occurs in the endometrium at the beginning of decidualization. This process seems to be associated with an increase in the endometrial expression of proteins such as RCAS1 that are involved in the suppression of immune cell activity. The aim of the present study was to evaluate alterations in the RCAS1 blood serum concentration levels in women with uterine leiomyoma over the course of the different menstrual cycle phases and to compare these levels with those found in patients suffering from adenomyosis. MATERIAL AND METHODS: The sRCAS1 blood serum concentration level was determined for 87 patients, including 38 patients with both adenomyosis and uterine leiomyoma, and 49 suffering from leiomyomatosis alone. RESULTS: Fluctuations in sRCAS1 blood serum concentration levels correlating with the menstrual cycle phases were demonstrated in patients suffering from uterine leiomyoma alone. The highest level of sRCAS1 concentration was found during the secretory cycle phase and the lowest during the proliferative cycle phase. However, no such fluctuations correlating with menstrual cycle phases were observed in patients suffering from both adenomyosis and leiomyoma. In fact, the level of sRCAS1 blood serum concentration in patients with adenomyosis remained almost constant. Patients with adenomyosis and leiomyoma were characterized by statistically significantly higher blood serum sRCAS1 levels during the proliferative cycle phase in comparison with the sRCAS1 blood serum levels in patients with leiomyoma alone. CONCLUSION: The lack of alterations in the sRCAS1 blood serum concentration levels observed in patients with adenomyosis may favor the development of the condition. PMID- 19122464 TI - Hepatocyte growth factor treatment improves alveolarization in a newborn murine model of bronchopulmonary dysplasia. AB - BACKGROUND: Bronchopulmonary dysplasia (BPD) is thought to be one form of developmental arrest of the lung. Hepatocyte growth factor (HGF) participates in normal lung growth and in lung regeneration. OBJECTIVES: The purpose of this study is to investigate whether HGF can improve alveolarization and attenuates functional abnormalities of a murine model of BPD induced by hyperoxia. METHODS: Three-day-old CD-1 mice were exposed to 90% of oxygen or room air (control group) for 7 days. These animals were then kept in room air for the next 7 days. Recombinant human (rh) HGF (100 microg/g b.w., divided 3 times, rhHGF group) or vehicle (vehicle group) was administered intraperitoneally during hyperoxia. On day 17, the pulmonary function test and bronchial hyperresponsiveness (BHR) were examined. Mean linear intercepts (MLI) were measured as parameters of alveolarization. Cell renewal (on day 10) and vascularization of the lung were also evaluated. RESULTS: Exposure to hyperoxia induced increased airway resistance and BHR. These animals showed a severely simplified alveolar structure, increased MLI, decreased cell renewal (16.1 +/- 2.4 vs. 29.6 +/- 2.4%, p < 0.05), and decreased vascularization (15.1 +/- 0.3 vs. 18.4 +/- 1.5 vessels/hpf, p < 0.05, vehicle vs. control group, respectively). rhHGF treatment during exposure to hyperoxia significantly reduced all of these changes (27.9 +/- 1.7%, 18.2 +/- 0.5 vessels/hpf for cell renewal and vascularization, respectively; all values are p < 0.05 against vehicle animals). CONCLUSION: HGF partially protects against the inhibition of alveolarization and improves functional abnormality in the hyperoxia-induced neonatal mice model of BPD. PMID- 19122465 TI - Germ cell tumors of the gonads: a selective review emphasizing problems in drug resistance and current therapy options. AB - Germ cell tumors are the most common malignancies in men under 50 years and also the most common cause of death from solid tumors in this age group. The past few decades have seen no change in the fact that 40% of such tumors have already metastasized by the time of initial diagnosis. In contrast to the majority of metastasized solid tumors, germ cell tumors can be effectively treated by cisplatin-based polychemotherapy. They only rarely show resistance mechanisms. Resistance to chemotherapeutics has been extensively examined in urological and nonurological tumors. Various mechanisms enabling malignant tumor cells to avoid the cytotoxic effect of chemotherapeutics have been discovered in the past few years. On the other hand, the reasons for the excellent responsiveness of germ cell tumors to chemotherapy have remained unclear. Theoretical models for the high cure rates of germ cell tumors after cisplatin-based polychemotherapy consider, on the one hand, molecular-biological aspects of malignant germ cell tumor cells per se and, on the other hand, the immunological and nonimmunological response mechanisms of the patients. This review article summarizes theoretical models for the high chemotherapy sensitivity of germ cell tumors and points out new therapeutic prospects. PMID- 19122466 TI - Erlotinib for metastatic non-small-cell lung cancer: first-, second- or third line setting - does it matter ? A single-institution experience. AB - BACKGROUND: Erlotinib is approved as treatment for metastatic non-small-cell lung cancer (NSCLC), following failure of initial therapy. Studies to define patients that derive maximal benefit from erlotinib have not dictated current practice. METHODS: We retrospectively analyzed the prescription patterns and outcomes related to erlotinib use for NSCLC in a comprehensive cancer center. RESULTS: Of 137 consecutive patients treated with erlotinib over 2 years, 116 were evaluable. Median age was 66 years, 63% females, most common histology was adenocarcinoma (n = 58). Seventy-nine patients presented with stage IIIB-IV disease, 37 with recurrent disease. There were 109 smokers. Erlotinib was given first line in 31 (27%), second line in 52 (45%) and third line in 33 (28%) patients. Daily erlotinib dose was 100 mg in 21 (18%) and 150 mg in 91 (82%) patients. Median duration of treatment was 8 weeks (range 1-72). Median overall survival (OS) from initiation of erlotinib was 5.4 months (range 0.2-27.8). There was no significant difference in median survival by disease stage (recurrent vs. de novo IIIB-IV) (p = 0.201), whether erlotinib was used as first-, second-, third-line therapy (p = 0.971) or at different doses (100 vs. 150 mg daily dose) (p = 0.579). CONCLUSIONS: OS after erlotinib use was not different, whether used as first-, second- or third-line therapy, whether patients had recurrent metastatic NSCLC or de novo stage IV disease, or if erlotinib was used at a dose of 100 or 150 mg daily. PMID- 19122468 TI - Role of cysteine in corneal wound healing after photorefractive keratectomy. AB - AIMS: To evaluate the efficacy of cysteine oral supplements in corneal wound healing after photorefractive keratectomy (PRK). METHODS: A total of 200 eyes (100 patients) underwent PRK. We divided patients into 2 groups; each group was composed of 50 patients (100 eyes). Group 1 patients were assigned to use oral L cysteine, with a daily dose of 200 mg (100-mg pills to be taken twice a day) for a duration of 14 days, starting from the week preceding the operation; the patients in group 2 were used as a control, and received the oral supplementation of 40-mg placebo pills to be taken twice a day for a duration of 14 days, starting from the week preceding the operation. RESULTS: All the eyes of patients treated with cysteine oral supplements showed shorter times to re-epithelization than the eyes of the control group. The mean time of corneal wound healing observed in group 1 was 102 +/- 15 h, compared to an average of 159 +/- 9 h in group 2. CONCLUSION: Our research showed that oral cysteine supplementation, in a daily dose of 200 mg, reduces mean corneal wound healing time in patients after PRK. PMID- 19122469 TI - Correlation of cell cycle kinetics with p63 expression in human limbal epithelial cells expanded on intact human amniotic membrane. AB - AIM: To analyse the correlation of p63 expression and cell cycle kinetics of human limbal epithelial cells (HLECs) expanded on amniotic membrane (AM) or plastic. METHODS: Primary HLECs were cultured either on cryopreserved intact AM or plastic dishes for 2 weeks. Cells were labelled with 5 microM 5-bromo-2' deoxyuridine (BrdU) for 3 days, followed by an interval of either 7 or 14 days in BrdU-free medium. The expression of p63 and BrdU labelling was detected by immunocytochemistry. RESULTS: More cells on AM (56%) than on plastic (24%) retained their BrdU label after the 7-day interval (p < 0.001). The difference was even more pronounced after 14 days (20 and 2%, p < 0.001). All BrdU-labelled cells were also p63 positive. 2.5-fold more cells on AM (54%) than on plastic (21%) were BrdU positive/p63 positive after 7 days (p < 0.001). It increased to a 9-fold difference after 14 days (p < 0.001). The BrdU label was lost more quickly than the p63 expression during the observation period, indicating that p63 expression was not confined to stem cells but existed also in transient amplifying cells. CONCLUSIONS: The combination of p63 expression and BrdU label retention is a better criterion to characterize stemness than either marker alone. AM as a culture substrate preserves stemness better than plastic. PMID- 19122470 TI - Ciliary body toxicity of subconjunctival suramin compared with mitomycin-C in the rabbit eye: determining the toxic concentration. AB - BACKGROUND/AIMS: To evaluate the safety of suramin compared with mitomycin-C (MMC) as an adjunctive agent in trabeculectomy by determining its ciliary body toxicity at predetermined effective dosages in rabbit eyes. METHODS: Thirty-two New Zealand albino rabbits received either suramin (200, 300, 400, or 800 mg/ml) or MMC (0.2, 0.3, 0.4, or 0.8 mg/ml) injections subconjunctivally in the right eye. Enucleations were performed on the 1st, 3rd, 7th and 28th day. Untreated left eyes were injected with balanced salt solution and served as controls. The injection-exposed ciliary body specimens were processed to be investigated under the light microscope and transmission electron microscope. RESULTS: There was no pathologic abnormality in specimens under light microscopy. The morphologic evaluation with transmission electron microscopy showed severe changes in structure, except for eyes treated with 200 mg/ml suramin and 0.2 mg/ml of MMC. These changes were more prominent in eyes exposed to MMC, and appeared earlier compared to suramin-treated eyes. CONCLUSIONS: Suramin 200 mg/ml and MMC 0.2 mg/ml seem to be comparatively nontoxic to the ciliary body of the rabbit eye. Concentrations higher than these values caused severe damage. PMID- 19122471 TI - Glyceraldehyde is present in rat lens and its level is increased in diabetes mellitus. AB - PURPOSE: OP-lysine, a glycation product of lysine residues of proteins, has been reported to be formed with glyceraldehyde and glycolaldehyde as precursors in the lens, and has been suggested to play a role in senile cataracts. However, there has been no reliable information regarding the content of glyceraldehyde in tissues. This study determined the glyceraldehyde levels in the lenses of normal and diabetic rats. METHODS: Glyceraldehyde was derivatized to a fluorescent compound, and the compound was then quantified by high-performance liquid chromatography. RESULTS: The lens glyceraldehyde levels in normal and diabetic rats were 0.75 +/- 0.06 and 1.26 +/- 0.21 nmol/g wet weight (means +/- standard deviations of 6 animals, p < 0.01), respectively. Isolated rat lenses accumulated a higher level of glyceraldehyde when cultured for 6 days in 25.5 mM glucose than when cultured in 5.5 mM glucose. CONCLUSIONS: Glyceraldehyde was found to be present in the lens and was increased in diabetes mellitus. OP-lysine is thus likely to be a potential risk factor for senile and diabetic cataracts. PMID- 19122467 TI - The role of antimicrobial peptides at the ocular surface. AB - Antimicrobial peptides (AMPs) such as defensins and cathelicidins are small peptides with broad-spectrum activity against bacteria, fungi and viruses. In addition, several AMPs modulate mammalian cell behaviours including migration, proliferation and cytokine production. This review describes findings from recent studies showing the presence of various AMPs at the human ocular surface and discusses their mechanism of antimicrobial action and potential non-microbicidal roles. Corneal and conjunctival epithelial cells produce beta-defensins and the cathelicidin LL-37, whereas neutrophils, infiltrating in response to a specific stimulus, supply additional LL-37 as well as alpha-defensins. In vitro studies suggest that LL-37 and human beta-defensin-3 are the most likely to have significant independent antimicrobial activity, while other AMPs may act synergistically to help protect the ocular surface from invading pathogens. Current evidence also supports a role for some AMPs in modulating wound healing responses. Although yet to be brought to fruition, AMPs hold significant potential as therapeutic agents for the prophylaxis and treatment of infection, promotion of wound healing and immune modulation. PMID- 19122472 TI - Aqueous humor concentrations of topically administered caspofungin in rabbits. AB - BACKGROUND: The echinocandin caspofungin (CAS) is a novel antifungal drug with fungicidal in vitro activity against all Candida spp., which are the most frequent cause of fungal keratitis. Penetration of CAS through the cornea into the aqueous humor after topical administration was investigated. METHODS: A CAS solution with a concentration of 7 mg/ml was applied onto each rabbit's cornea. Drug application after corneal epithelium abrasion was processed in different time intervals: single application with aqueous humor sampling after 1 and 2 h. In addition, after continuous application of CAS every 30 min, aqueous humor concentrations of CAS after 1, 2 and 5 h were analyzed by liquid-chromatography tandem mass spectrometry. RESULTS: Topical administration of CAS without corneal epithelium abrasion resulted in no detectable amounts of the drug in the aqueous humor. However, with corneal abrasion, after a single application, levels of 2.16 +/- 1.57 microg/ml (n = 6) were reached after 1 h and then decreased to 1.76 +/- 0.88 microg/ml (n = 2) after 2 h. After serial application every 30 min, the following intracameral levels of CAS were detected: after 1 h, 2.11 +/- 1.09 microg/ml (n = 6); after 2 h, 4.94 +/- 1.80 microg/ml (n = 5), and after 5 h, 3.45 +/- 2.11 microg/ml (n = 6). CONCLUSION: In the aqueous humor, therapeutic drug levels can be reached that cover the MICs of most fungi after epithelial abrasion. To achieve a sustained high level of CAS as an effective antifungal therapy for corneal keratitis, CAS should be administered topically every 30 min after removal of the corneal epithelium. PMID- 19122473 TI - Immunohistochemical localization of fibrillin-1 protein in the cells of chick corneal and conjunctival epithelia during pre- and postnatal development. AB - Fibrillin-1 protein is a microfibrillar glycoprotein component of the extracellular matrix, widely distributed in ocular connective tissues. In this work, we show for the first time the expression pattern of fibrillin-1 protein in the corneal and conjunctival epithelia and in stromal keratocytes during embryo development. After hatching, protein expression was maintained in the corneal epithelium cells and nonsecreting epithelium cells of the conjunctiva and disappeared in the stromal keratocytes. In the limbus region, the basal cells were negative, while superficial cells were positive for the antibody. The expression in corneal epithelial cells suggests a role for fibrillin in development and disease. Therefore, some basal cells of the limbus region do not show fibrillin-1 immunolocalization, and this may be correlated with stem cell or stem-like properties. PMID- 19122474 TI - Immunosuppressive property of dried human amniotic membrane. AB - PURPOSE: To report the immunosuppressive property of dry human amniotic membrane (dHAM). METHODS: Mouse splenocytes harvested from Balb/c mice were stimulated using functional-grade anti-CD3e antibodies for 4 days either with or without either of 2 commercial types of dHAM (Ambiodry 1 & 2, IOP Inc., Costa Mesa, Calif., USA) added to the culture media. The cell proliferation assay was performed to analyze the extent of splenocyte proliferation. RESULTS: dHAMs significantly suppressed mouse splenocyte proliferation compared to control. The suppression by dHAM with intact amniotic epithelium (Ambiodry 2) was significantly stronger than dHAM without epithelium (Ambiodry 1). CONCLUSION: The immunosuppressive property of dHAM was demonstrated using an alloreactive splenocyte proliferation assay. PMID- 19122475 TI - Potential use of riboflavin/UVA cross-linking in bullous keratopathy. AB - BACKGROUND: Collagen cross-linking of the cornea has been shown by us to have an antiedematous effect in the cornea. The aim of the present study was to examine if this effect can be used for the treatment of bullous keratopathy. METHODS: This clinical interventional case series included 3 patients (3 eyes) with bullous keratopathy due to pseudophakia, corneal transplant rejection, and Fuchs' endothelial dystrophy. After dehydration for 1 day using 40% glucose, the central 8 mm of the cornea were abraded and cross-linked with the photosensitizer riboflavin and UVA (370 nm, 3 mW/cm(2)) for 30 min. Optical coherence tomography pachymetry measurements of the central cornea were performed at various time intervals. RESULTS: Corneal thickness was reduced by 90.33 +/- 17.04 microm on average 3 days after cross-linking and by 93.67 +/- 14.22 microm after 8 months. The bullous changes of the epithelium were markedly improved, resulting in loss of pain and discomfort. Visual acuity was significantly improved in the case without prior stromal scarring. CONCLUSIONS: Cross-linking might become another useful tool in the treatment of bullous keratopathy. It is primarily suited for patients with pain symptoms, restricted visual prognosis or to extend the time interval for an upcoming corneal transplantation. PMID- 19122476 TI - Bioequivalence of HX575 (recombinant human epoetin alfa) and a comparator epoetin alfa after multiple subcutaneous administrations. AB - AIM: To compare the steady-state pharmacokinetics and pharmacodynamics (PK/PD) of two erythropoesis-stimulating agents (ESA), HX575 (Binocrit, Sandoz GmbH, Holzkirchen, Germany), human recombinant epoetin alfa approved as the first biosimilar ESA, and a comparator epoetin alfa, following multiple subcutaneous administrations. METHODS: An open, randomized, parallel group study was conducted in 80 healthy adult males. Subjects were randomized to multiple subcutaneous doses of 100 IU/kg body weight of HX575 or of the comparator epoetin alfa 3 times weekly for 4 weeks. RESULTS: The hematological profiles of both treatments were similar, as determined from the population mean curves and area under the effect curve (AUEC) ratios. HX575 met the predefined biosimilarity criteria with respect to the ratio and 90% confidence interval of the AUEC(Hb) (98.9% [97.7-100.2%]), the primary PD endpoint. The PK of the two treatments were also similar as shown by the AUC(0-48) ratios and 90% confidence intervals, 94.3% [84.7-105.0%] and 96.9% [88.2-106.5%], respectively. Study medication was well tolerated and neutralizing anti-epoetin antibodies were not detected. CONCLUSIONS: HX575 and the comparator epoetin alfa were bioequivalent with respect to their PK/PD, supporting the conclusion that both, when administered subcutaneously, will be equally efficacious and may be interchangeable as therapy. PMID- 19122477 TI - Load-induced cardiomyocyte apoptosis in cultured multicellular myocardial preparations is unaltered in presence of the beta-adrenoceptor antagonist nebivolol. AB - Overload-induced heart failure is associated with enhanced apoptosis of cardiomyocytes, and increased mechanical load is an inductor of this apoptosis. It is unknown whether nebivolol, a third generation beta(1)-adrenoceptor antagonist, possesses properties that can attenuate this apoptosis. Multicellular preparations from rabbit hearts were mounted in a culture system that allows for measurement of contractile parameters over several days. Culturing these muscles on a constant high preload induces apoptosis of the cardiomyocytes. Of each heart, 1 preloaded muscle preparation was treated with nebivolol (10(-6) mol/l), 1 preloaded without continuous exposure to nebivolol (positive control) and 1 unloaded (negative control). After 48 h of continuous loaded contractions, apoptosis was assessed by TUNEL-assay to confirm that nuclei of myocytes were affected, or by DNA-ladder intensity analysis for semiquantification. Maximal twitch force development was slightly, but significantly, lower in preparations contracting in presence of nebivolol (compared to solvent) while twitch-timing parameters were similar. After 48 h of continuous contractions, no additional differences were observed between the groups regarding contractile parameters. DNA-ladder analysis showed a similar rate of apoptosis in presence of nebivolol. Nebivolol does not increase, nor decrease, the rate of load-induced cardiomyocyte apoptosis. PMID- 19122478 TI - Drug rash with eosinophilia and systemic symptoms after chlorambucil treatment in chronic lymphocytic leukaemia. AB - Drug rash with eosinophilia and systemic symptoms (DRESS) is a severe cutaneous drug reaction with a long duration of eruption and serious organ involvement. The mortality rate has been estimated at about 10%. Aromatic anticonvulsants, sulphamides, minocycline and more rarely carbamazepine are the principal responsible drugs. We report the first case of chlorambucil-induced DRESS syndrome in a 70-year-old man recently diagnosed with chronic lymphocytic leukaemia. He developed recurrent skin rash, fever, hypereosinophilia, and acute renal failure after rechallenge with chlorambucil. The condition improved slowly after stopping medication and systemic steroids. Prompt recognition of a chlorambucil drug reaction is essential in patients receiving chemotherapy. PMID- 19122479 TI - The missing link--light-induced (280-1,600 nm) free radical formation in human skin. AB - The recent European recommendation on the efficacy of sunscreen products requests now a minimum ratio of UVA/UVB protection. However, the visible and the infrared (IR) parts of the sun spectrum have received little attention concerning their possible contribution to skin damage. A common biophysical answer for the different wavelengths of the sun spectrum can be found in the creation of excess free radicals - mainly reactive oxygen species (ROS). Thanks to electron spin resonance spectroscopy applied to skin biopsies, we determined for the first time the free radical action spectrum covering UV and visible light (280-700 nm). Convolution of the action spectrum with sunlight spectral irradiance showed that 50% of the total skin oxidative burden was generated by visible light. Creation of ROS by visible light was experimentally confirmed by varying the illuminance of a spotlight. We also evidenced the creation of excess free radicals by near-IR radiation. In that case, free radical generation does not depend exclusively on the dose, but also on the skin temperature increase initiated by near-IR light. Some phenomena which are still unclear, such as the question about the deleterious or beneficial role of sunlight, are reviewed, implying the research on new protection strategies for the prevention of skin cancer. PMID- 19122480 TI - [Development of a structural equation model to estimate university students' depression]. AB - PURPOSE: This study was designed to construct a structural model explaining depression in university students. METHODS: Data were collected from 1,640 university students by questionnaire, and analyzed using AMOS 5.0 to test the hypothetical model. RESULTS: Fitness statistics for the modified model were GFI=.93, AGFI=.89, NFI=.91, and RMSEA=.081. All the 12 paths in the modified model proved to be statistically significant. Depression of university students accounted for 52% of the covariance by the factors. The factor that had the most influence on depression was individual vulnerability, and followed by sequence order, stress, social support, coping, and self-efficacy. Depression was influenced directly by individual vulnerability, stress, social support, and coping, and indirectly by individual vulnerability, stress, social support, and self-efficacy. CONCLUSION: A screening and management system for the high risk group is needed to effectively prevent depression and reduce rate of depression in university students. Detailed support programs which specifically deal with prevailing stressors should be developed to effectively reduce the harmful effects of individual vulnerability and stress. It is anticipated that the model constructed in this study could be utilized as a reference in developing various strategies to prevent and intervene depression in university students. PMID- 19122481 TI - [Case management process identified from experience of nurse case managers]. AB - PURPOSE: The purpose of this study was to develop a substantive theory of case management (CM) practice by investigating the experience of nurse case managers caring for Medical Aid enrollees in Korea. METHODS: A total of 12 nurses were interviewed regarding their own experience in CM practice. Data were recorded and analyzed using grounded theory. RESULTS: Empowerment was the core category of CM for Medical Aid enrollees. The case managers engaged in five phases as follows, phase of inquiring in advance, building a relationship with the client, giving the client critical mind, facilitating positive changes in the client's use of healthcare services, and maintaining relationship bonds. These phases moved gradually and were circular if necessary. Also, they were accelerated or slowed depending on factors including clients' characteristics, case managers' competency level, families' support level, and availability of community resources. CONCLUSION: This study helps understand what CM practice is and how nurses are performing this innovative CM role. It is recommended that nurse leaders and policy makers integrate empowerment as a core category and the five critical CM phases into future CM programs. PMID- 19122482 TI - [The lived experience of struggling against illness for patients with amyotrophic lateral sclerosis]. AB - PURPOSE: The purpose of this study was to identify and describe phenomenological structures of the lived experience of struggling against an illness for patients with Amyotrophic Lateral Sclerosis (ALS). METHODS: The participants were 7 patients with ALS recruited by snowball sampling who agreed to participate in this research and could verbally communicated with the researcher. Data were collected by long term-repeated interviews with participants in their own homes. Data were analyzed using Colaizzi's method of phenomenology. RESULTS: Four categories were extracted as follows: 'Being seized with fear of death', 'Living a marginal life', 'Accepting hard fate', and 'Clinging to faint life'. Seven theme clusters were identified as: 'Wandering to find a healing method with ominous signs in the body', 'Having a diagnosis of ALS is like a bolt from the blue and struggling against illness with faint hope', 'Being forced out to the edge of life with anguish', 'Filling one's heart with hatred and longing toward becoming estranged from the world', 'Living with stigma as a stumbling block with bitter grief in one's heart', 'Accepting every things as one's fate with self controlled fear of death', and 'Attaching to desire to live'. CONCLUSION: The results of this study can be used to develop the programs to support patients with ALS and their family. PMID- 19122483 TI - [Development of an osteoporosis awareness scale for women]. AB - PURPOSE: This study was conducted to develop an instrument measuring awareness of osteoporosis. METHODS: The process included construction of a conceptual framework, generation of 86 preliminary items, verification of content validity, selection of secondary items, verification of construct validity and extraction of final items. The 86 items were reviewed for content validity by two groups of experts and were tested to evaluate inter item correlation coefficient by two groups of adult women. From June to August 2007, data were collected from 383 adult women who lived in Seoul and provinces in Korea. Data were analyzed by performing item analysis, Varimax factor analysis and Cronbach's alpha. RESULTS: There were 31 items in the final instrument categorized into 5 factors. The factors were labeled as "preventive behaviors (10 items)", "risk factors (5 items)", "characteristics of osteoporosis (6 items)", "improving bone health (5 items)", and "bone physiology (5 items)". Cumulative percent of variance was 60.92% and eigen values ranged from 1.20 to 12.44. Cronbach's alpha was .948 and ranged from .804 to .917. CONCLUSION: Validity and reliability of the scale are confirmed in this study showing its utility for measuring awareness of osteoporosis for women. Utilization of the scale will also contribute to designing an appropriate prevention program for osteoporosis. PMID- 19122484 TI - [Effects of community-based case management program for clients with hypertension]. AB - PURPOSE: The purpose of this study was to analyze effects of a community-based case management program for clients with hypertension living in the community. METHODS: The research design was a one group pre and post-test design with 30 participants with hypertension who agreed to participate in the 8-12 week case management program provided by case managers from the National Health Insurance Corporation in 2002. Data were collected three times, before and after the case management services, and 6 months later. Outcomes included changes in blood pressure, knowledge of hypertension and daily life practices, including alcohol consumption, smoking, exercise, and medication adherence. RESULTS: Repeated measures ANOVA and post-hoc tests of means revealed significant differences before and after service for systolic blood pressure, daily life practices (monitoring body weight and BP, low salt and cholesterol and high vegetable diet, and stress-relief practices), and exercise. The goal for medication adherence was attained after service. Significant improvements from baseline to 6 months after service were observed in measures of salt and vegetables in diet. There were no significant differences on hypertension knowledge, alcohol consumption or smoking behavior between before service and after, and at 6 months. CONCLUSION: The findings provide preliminary evidence that case management intervention can have positive outcomes on BP control, daily life practices, exercise, and medication adherence for clients with hypertension. However, additional interventions are needed to sustain long-term effects. PMID- 19122485 TI - [Development of a web-based senescence preparation education program for successful aging for middle-aged adults]. AB - PURPOSE: The purpose of this study was to develop a web-based senescence preparation education program to promote successful aging. METHODS: This program was developed based on Network-Based Instructional System Design (NBISD) model, using the following 5 processes: analysis, design, development, implementation, and evaluation. The program was operated for 10 weeks from March 17 to May 25, 2008. RESULTS: There were 4 menu bars, introduction, related data, lecture room, and communication on the main page. In the operation of this program, HTML, ASP, JAVA Script, Namo web editor, Edit Plus, Front Page and multimedia technology were applied. The program content consisted of understanding elderly people, physical health, activity & exercise, nutrition, medication use, psychological health, intellectual health, understanding death, welfare system and leisure activity. CONCLUSION: This program could be a useful means to provide senescence preparation information to middle-aged adults. Also, it is expected to offer individualized learning opportunities to many learners in various settings. Nurses should further develop and facilitate various learning strategies including web- based programs for elder care. PMID- 19122486 TI - [Effects of a breast feeding promotion program for working women]. AB - PURPOSE: The purpose of this study was to measure the effects of a breast feeding promotion program for working women on breast feeding continuation, mother-infant attachment, and maternal sensitivity. METHODS: The design of this study is nonequivalent control group design with repeated measures. RESULTS: There were significant differences in breast feeding continuation between two groups at each time point except 2 days and 1 week after delivery. The scores of mother-infant attachment and maternal sensitivity of experimental group were higher than those of the control group, but there were no significant differences between two groups. CONCLUSION: Although some modifications in contents and administration will be required to increase the effectiveness of the program, breast feeding promotion program for working women can be an effective nursing intervention which can facilitate breast feeding continuation and mother-infant relationships. PMID- 19122487 TI - [Task analysis of the job description of gerontological nurse practitioners based on DACUM]. AB - PURPOSE: The aim of this study was to develop and to analyze the task of gerontological nurse practitioners (GNPs) in Korea. METHODS: The definition of GNP and job description was developed based on developing a curriculum (DACUM) by 7 panels who have experienced in DACUM analysis and gerontological nursing. One hundred sixty nurses who were working at long term care facilities were participated. The questionnaire included frequency, importance, and difficulty of duties, tasks, and task elements. The data were collected in November 2006, analyzed by descriptive statistics. RESULTS: The job description of GNPs in Korea revealed 5 duties, 23 tasks, and 86 task elements. On the all five duties, the highest duty in frequency and in importance was professional nursing care (3.25+/ 0.35, 3.49+/-0.29). But the highest duty in difficulty was research (3.24+/ 0.46). 'Prevent health problem (3.42+/-0.43, 3.56+/-0.33)', 'Teach other staffs (2.83+/-0.77, 3.39+/-0.43)', 'Develop the evidence-based standards (2.43+/-0.76, 3.22+/-0.43)', 'Develop the self (2.81+/-0.65, 3.26+/-0.42)', and 'Participate the team activities' were the highest score in frequency and in criticality of tasks. 'Provide emotional support to older adults and families (3.16+/-0.41)', 'Counsel older adults and their families (3.14+/-0.49)', 'Do clinical research (3.32+/-0.49)', 'Quality insurance (3.25+/-0.49)', and 'Build collaborative system (3.18+/-0.47)' were perceived the most difficult tasks. CONCLUSION: The political efforts for the legislation of role and task of GNPs were needed. PMID- 19122488 TI - [Factors explaining quality of life in individuals with coronary artery disease]. AB - PURPOSE: The study was done to compare quality of life by gender, and to identify factors which explain quality of life in individuals with coronary artery disease. METHODS: For the survey, 91 individuals (53 men and 38 women) agreed to participate in the study. Cardiovascular risk factors, systolic blood pressure, body mass index, total cholesterol, triglyceride, high density lipoprotein cholesterol, and low density lipoprotein-cholesterol, health behavior as well as quality of life, were measured. Descriptive statistics, t-test, correlation and hierarchical multiple regression with SPSS WIN 12.0 were used to analyze the data. RESULTS: Significant gender differences were found for education, smoking status, chronic disease, perceived health status, and quality of life within sub dimensions. Hierarchical regression analysis showed gender (men), age, perceived health status, cardiovascular risk scores, and health behaviors together explained 40.2% (adjusted R2) of variance in quality of life. CONCLUSION: As the factors explaining quality of life in individuals with coronary artery disease have been identified as gender (men), age, perceived health status, and health behaviors, health promotion programs designed for this population should focus on these factors for effective behavioral modification, and consequent improvement in quality of life. PMID- 19122489 TI - [Study on the changes in limb circumferences and muscle areas of critically ill patients using ventilators]. AB - PURPOSE: The purpose of this study was to identify muscle atrophy in critically ill patients on ventilators. A comparison was made between limb circumferences and muscle areas on the day of admission to an intensive care unit (ICU) and on the 8th day after admission. METHODS: The data were collected from 30 critically ill patients using ventilators at S hospital ICU in Seoul during the period from October 2005 to April 2006. Limb circumferences and skinfold thickness were measured on the day of admission and on the 8th day after admission to the ICU. Limb circumferences and skinfold thickness were measured on the right mid-arm, right mid-thigh, and right mid-calf using a measuring tape and a skinfold calipers. Limb muscle areas were calculated by an equation after measuring limb circumferences and skinfold thickness. The data were analyzed by paired t-test and independent t-test. RESULTS: 1) Mid-arm circumference, mid-thigh circumference and mid-calf circumference on the 8th day after admission to the ICU were significantly less than those on the day of admission to the ICU, 2) Mid arm muscle area, mid-thigh muscle area and mid-calf muscle area on the 8th day after admission to the ICU were significantly less than those on the day of admission to the ICU, 3) Steroid and vecuronium medication didn't affect changes in limb circumferences (mid-arm, mid-thigh, mid-calf) and muscle areas (mid-arm, mid-thigh, mid-calf) on the 8th day after admission to the ICU compared to the day of admission. CONCLUSION: Limb muscle atrophy may occur on the 8th day after admission to the ICU in critically ill patients using ventilator. PMID- 19122491 TI - [The effects of an environmental education with newspaper in education (NIE) on the environmental concern and practice]. AB - PURPOSE: The purpose of this study was to develop an environmental education program using newspaper articles in education (NIE) and to evaluate changes in concern and practice for environmental protection after NIE. METHODS: The design was a nonequivalent control group pretest-posttest design. The participants were university students in nursing, of which 31 were assigned to the experimental group and 43 to the control group. The education was carried out for 2 hr, once a week for 7 weeks. Data were analyzed with SPSS WIN 14 program, and included chi2 test, independent t-test, and repeated measure ANOVA. RESULTS: NIE showed significant differences in the changes of attitude toward environment (F=4.461, p=.036). CONCLUSION: Findings suggest that this NIE in environmental education was effective in changing students' attitudes toward the environment. Therefore this NIE is recommended for inclusion in education for university students in nursing. PMID- 19122490 TI - [Psychometric evaluation of the Korean social support questionnaire]. AB - PURPOSE: Research related to social support in Korea has been hampered by paucity of measurement tools reflecting Korean culture. The aim of the study was to develop Korean social support questionnaire (KSSQ) based on the Korean social support pyramid and to test psychometric properties of the KSSQ. METHODS: The questionnaire was administered to 701 subjects and 658 college students. Psychometric analyses included factor analyses, expert validity, criterion related validity, internal consistency, and test-retest reliability. RESULTS: A principal components analysis support for construct validity, eliciting a three factor solution accounting for 65.46% of variance in scores. Concurrent and discriminant validity supported criterion-related validity. Internal consistency of reliability was support with Cronbach's alpha of .97-.98 for the entire scale. Test-retest reliability was .76. CONCLUSION: This initial testing of KSSQ to measure Korean social support demonstrates evidence of reliability and validity. Assessment of known-group validity and norm establishment of KSSQ are suggested to provide further sound psychometric properties and practical measurement tools. PMID- 19122492 TI - [Effects of a strengthening program for lower back in older women with chronic low back pain]. AB - PURPOSE: The purpose of this study was to identify effects of a strengthening program for the lower back in older women with chronic low back pain. METHODS: The research design was a nonequivalent control group pretest-posttest experiment. The experimental group consisted of 16 older women and the control group, 14, all of whom had experienced low back pain for at least 3 months. The strengthening program for the lower back included lumbar stabilization exercises and education on pain management in daily living. For an 8 week period, exercises were done 3 days a week and on one day education was also given. RESULTS: Pain and disability scores decreased significantly in the experimental group compared to the control group. Flexibility, life satisfaction and lumbar muscle strength scores increased significantly in the experimental group compared to the control group. CONCLUSION: Low back pain and disability can be relieved, and flexibility, muscle strength, and life satisfaction increased through a program to strengthen the lower back. It is suggested that a program to strengthen the lower back would be an effective nursing intervention for older women with low back pain. PMID- 19122493 TI - [Effect of a tooth-brushing education program on oral health of preschool children]. AB - PURPOSE: To examine the effect of tooth-brushing education on the oral health of preschoolers. METHODS: A quasi-experimental design with a non-equivalent control group was used. Two kindergartens were selected and 39 preschoolers from one kindergarten were assigned to the experimental group with tooth-brushing education and 39 from the other kindergarten to the control group. The tooth brushing education program included 1 session on oral health education, individual tooth-brushing instruction for 1 week and supervised tooth-brushing after lunch for 4 weeks. Oral health behavior including use of tooth paste, tooth brushing time and method of tooth-brushing, plague, streptococcus mutans, lactobacillus and dental caries were measured before and after the education. Fisher's exact test, t-test and paired t-test with the Window SAS 9.1 program were used to analyze the data. RESULTS: A significant increase in the use of tooth paste, tooth-brushing time and the practice of correct tooth-brushing and a decrease in plague and development of dental caries were observed in the experimental group. CONCLUSION: This tooth-brushing education was partially effective in improving oral health of preschoolers. PMID- 19122494 TI - [Influence of age, gender, and severity on recovery of patients with brain injury]. AB - PURPOSE: This study was conducted to investigate the individual and cross influences of age, gender, and severity on recovery of patients with brain injury. METHODS: For the purpose of the study, traumatic or spontaneous brain injury patients admitted to the intensive care unit (ICU) were conveniently selected. The data regarding outcomes were collected 3 months after admission. RESULTS: Individual influences of the study variables on patients' recovery were significant, except for gender. But while the individual influence of gender on recovery was not significant, cross influence of gender and age was significant, but only for the sub-dimension of 'arousalbility and awareness'. The study results also showed that 3-way cross influence of gender, age, and severity was only significant on the sub-dimension of 'arousalbility and awareness'. CONCLUSION: The sub-dimension of recovery cross influenced by the demographic factors of gender and age, and severity was 'arousalbility and awareness'. This might indicate that the study variables that cross influencing recovery had more influence on consciousness compared to physical function and psycho-social adaptation. PMID- 19122496 TI - Pancreas. PMID- 19122497 TI - Receptor-mediated signal transduction pathways and the regulation of pancreatic acinar cell function. AB - PURPOSE OF REVIEW: Recent studies on pancreatic acinar cell function have led to a more detailed understanding of the signal transduction mechanisms regulating digestive enzyme synthesis and secretion as well as pancreatic growth. This review identifies and puts into context these recent studies, which further understanding in these areas. RECENT FINDINGS: Receptors present on acinar cells, particularly those for cholecystokinin and secretin, have been better characterized as to the molecular nature of the ligand-receptor interaction. Other reports have described the receptors for natriuretic peptides and fibroblast growth factor on acini. Intracellular Ca(2+) signaling remains at the center of stimulus secretion coupling and its regulation by inositol 1,4,5 trisphosphate, nicotinic acid adenine dinucleotide phosphate and cyclic ADP ribose has been further defined. Work downstream of intracellular mediators has focused on molecular mechanisms of exocytosis particularly involving small G proteins, soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins and cytoskeletal proteins. Considerable progress has been made defining the complex in acinar cells and its regulation. In addition to secretion, recent studies have further defined the regulation of pancreatic growth both in adaptive regulation to diet and hormones, particularly cholecystokinin, and in the regeneration that occurs after pancreatitis or partial pancreatectomy. This regulation involves calcineurin-nuclear factor of activated T cells, mammalian target of rapamycin, mitogen-activated protein kinase, Notch signaling pathways as well as various tyrosine kinases. SUMMARY: Understanding the mechanisms that regulate pancreatic acinar cell function is contributing to our knowledge of normal pancreatic function and alterations in diseases such as pancreatitis and pancreatic cancer. PMID- 19122499 TI - Chronic pancreatitis. AB - PURPOSE OF REVIEW: As in previous reviews in this journal, this review is focused on the most important new observations in chronic pancreatitis made in the last year. RECENT FINDINGS: Important observations included the search for biomarkers and alternative methods for the detection of early chronic pancreatitis; stellate cell activation and their role in fibrogenesis; the natural history of chronic pancreatitis; reports outlining the complexity in diagnosis of autoimmune pancreatitis; emerging roles of endoscopic ultrasound and magnetic resonance cholangiopancreatography in chronic pancreatitis diagnosis; a better understanding of neurobiology of chronic pancreatitis pain; and the potential role of surgery as first-line therapy in advanced chronic pancreatitis. SUMMARY: In 2007, major advances were made in our understanding of central processing in chronic pancreatitis pain. New techniques are being utilized in search of a better means to diagnose early chronic pancreatitis. Important prospective studies are emerging, which compare endoscopic and surgical interventions. Furthermore, the complexities of diagnosing autoimmune pancreatitis are being recognized. Overall, the future is promising as advances in genomic and proteomic techniques are applied to improve our understanding of chronic pancreatitis. PMID- 19122500 TI - Autoimmune pancreatitis. AB - PURPOSE OF REVIEW: Autoimmune pancreatitis (AIP) is an increasingly recognized clinical condition. Our objective is to provide a concise review of the advances in the past year in our understanding of AIP. RECENT FINDINGS: In a hospital survey from Japan, the prevalence of AIP was estimated at 0.82 per 100,000 individuals. The pathogenesis of AIP remains unclear but a recent report noted that T helper type 2 and T regulatory cells predominantly mediate the immune reaction in AIP. Genetic associations that may predispose to relapse of AIP were reported. Multiple case series further described the clinical profile of AIP and its extrapancreatic manifestations. A large series on immunoglobulin G4 (IgG4) associated cholangitis noted that patients with IgG4-associated cholangitis presented with obstructive jaundice and had increased serum IgG4 levels and IgG4 positive cells in bile duct biopsy specimens. Tissue IgG4 staining is likely to be a useful adjunct to serological diagnosis. AIP is steroid-responsive but maintaining remission continues to remain challenging. Presently low-dose steroids or immunomodulators are being used but efficacy of these medications remains to be determined. SUMMARY: There has been significant progress in understanding the clinical profile of AIP but knowledge of pathogenesis remains limited. Treatment practices vary widely and management of refractory disease continues to be challenging. PMID- 19122498 TI - Molecular basis for pancreatitis. AB - PURPOSE OF REVIEW: This timely review will focus on clinical and basic science studies that have greatly advanced our knowledge of the molecular mechanisms of both acute pancreatitis and chronic pancreatitis over the last year. RECENT FINDINGS: Animal models of both severe acute pancreatitis and chronic pancreatitis have recently been developed. Several unexpected protective mechanisms, mediated by the protease activated receptor 2 and heat shock protein 70, have been described. A genetic study suggested that polymorphisms in toll like receptor-4 might affect the risk of developing infections in acute pancreatitis. Studies of chronic pancreatitis have shown that specific neural receptors, transient receptor potential vanilloid subtype 1, mediate pain responses in a model of chronic pancreatitis. The pancreatic zymogen, chymotrypsin C, can degrade pathologically activated trypsin in the acinar cell. Inactivating mutations in chymotrypsin C have been reported to predispose to the development of chronic pancreatitis, especially in those who are prone to alcohol abuse. SUMMARY: The implications of the last year's findings are widespread. Improved animal models of acute pancreatitis and chronic pancreatitis will be critical for performing pilot studies of therapy. A greater understanding of genetic factors and pain responses could lead to potential treatments. This review will first discuss issues related to acute pancreatitis, and then conclude with studies most relevant to chronic disease. PMID- 19122501 TI - The sunset of somatic genetics and the dawn of epigenetics: a new frontier in pancreatic cancer research. AB - PURPOSE OF REVIEW: The excitement of finding a cancer modulator which is either mutated or deleted in vivo (genetics), unfortunately, is shadowed by the fact that we scientists have failed to live to the promise of gene therapy, and therefore, these genes cannot be replaced to cure the patients. On the contrary, both DNA methylation and chromatin-mediated inactivation of tumor suppressor genes (epigenetics), for example, are reversible as demonstrated by the relative success of emerging therapies. Therefore, epigenetics with its molecular basis (DNA methylation and chromatin modification) is among the most promising areas of cancer research and is a nascent field in pancreatic cancer research. RECENT FINDINGS: Here, we review and update novel findings on epigenetics as it applies to pancreatic cancer. SUMMARY: Special focus has been given to novel potential therapeutic targets and currently available drugs, which are emerging from this exciting new field of pancreatic cancer research. PMID- 19122502 TI - Pancreatic surgery. AB - PURPOSE OF REVIEW: Pancreatic surgery is a very challenging field and the management of pancreatic diseases continues to evolve. This report reviews the most recent information relating to pancreatic surgery. RECENT FINDINGS: Startlingly, it appears that a third of patients with resectable pancreatic cancer do not receive an operation in the United States, but for those who do receive surgery new emphasis on adequate lymph node staging has been reported. Assessment of guidelines for the treatment of intraductal papillary mucinous neoplasm and cystic lesions of the pancreas appear to validate the current practice. Extended pancreatic resections with vascular resection appear safe and with reasonable survival outcomes. The use of laparoscopic techniques for the pancreas continues to advance. Finally, patients with chronic pancreatitis appear better served by operative techniques rather than endoscopic approaches. SUMMARY: Pancreatic surgery is a fascinating field as we learn more about the biology of the conditions that afflict this gland and the best practices to address these diseases. PMID- 19122503 TI - Argon plasma coagulation therapy. AB - PURPOSE OF REVIEW: To give a summary of recent advances in argon plasma coagulation (APC) therapy in gastroenterology. RECENT FINDINGS: APC has gained importance in gastrointestinal endoscopy during the last decade, and APC technology has improved over time. Recently published studies, mainly consisting of case series, have shown that APC can be used effectively in various forms of gastrointestinal disorders. The complication rates observed during clinical application are low. SUMMARY: APC can be considered as a well tolerated and effective treatment approach in endoscopy. Further clinical trials including a comparison of APC to other treatment modalities as well as long-term follow-up after APC treatment are required to clarify the relative value of APC in gastroenterology. PMID- 19122504 TI - Endoscopic papillectomy. AB - PURPOSE OF REVIEW: Although few controlled trials exist in the field, endoscopic papillectomy has evolved over the recent years with new data on preoperative staging and improved methods for its safe and successful completion. In 2006, a consensus guideline was published by the American Society of Gastrointestinal Endoscopy evaluating the role of endoscopy in managing ampullary adenomas. RECENT FINDINGS: The recent literature of endoscopic papillectomy has focused on the preoperative management of ampullary tumors, with a paper evaluating the role of endoscopic ultrasound. Also, a randomized controlled trial has shown that the use of pancreatic duct stents is associated with less incidence of postendoscopic papillectomy pancreatitis, although the study was probably underpowered. Several methods can be used to help locate the pancreatic duct postendoscopic papillectomy (endoscopic ultrasound-guided rendezvous and methylene blue injection). The recurrence and complication rate in more recent papers continue to be acceptable, at about 30 and 20%, respectively. SUMMARY: Endoscopic papillectomy is a reasonable alternative to transduodenal surgical excision, but more controlled studies with long-term data are needed to evaluate preoperative staging accuracy and recurrence rates. PMID- 19122505 TI - Endoscopic diagnosis and staging of mucosa-associated lymphoid tissue lymphoma. AB - PURPOSE OF REVIEW: The role of endoscopy, including endoscopic ultrasound, in the diagnosis and management of mucosa-associated lymphoid tissue lymphomas of the stomach has evolved steadily in the last two decades. The present review summarizes recent findings and puts them in context with studies on the diagnosis and management of mucosa-associated lymphoid tissue lymphoma published earlier. RECENT FINDINGS: Several recent studies have emphasized the crucial role of endoscopic ultrasound in treatment planning in patients with gastric mucosa associated lymphoid tissue lymphoma. This is important as early-stage gastric mucosa-associated lymphoid tissue lymphomas can be managed just by the eradication of Helicobacter pylori by appropriate antibiotic regimens. However, the more advanced lesions are treated with much more invasive treatment regimens involving radical gastrectomy, chemotherapy or radiation, or all. SUMMARY: Endoscopic ultrasound staging is highly accurate in predicting response to Helicobacter pylori eradication in patients with gastric mucosa-associated lymphoid tissue lymphoma. Normalization of gastric wall thickness and architecture can be used to monitor tumor regression following treatment. Endoscopic ultrasound findings can also be used to identify treatment failures and relapses and can help identify patients who need more aggressive therapy. PMID- 19122506 TI - Endoscopic tissue diagnosis of cholangiocarcinoma. AB - PURPOSE OF REVIEW: The extremely poor outcome in patients with cholangiocarcinoma, in large part, reflects the late presentation of these tumors and the challenging nature of establishing a tissue diagnosis. Establishing a diagnosis of cholangiocarcinoma requires obtaining evidence of malignancy from sampling of the epithelium of the biliary tract, which has proven to be challenging. Although endoscopic ultrasound-guided fine needle aspiration performs slightly better than endoscopic retrograde cholangiopancreatography in diagnosing cholangiocarcinoma, both endoscopic approaches demonstrate disappointing performance characteristics. RECENT FINDINGS: The limitations of cytologic analysis have recently led to the evaluation of new technologies such as digital image analysis and fluorescence in-situ hybridization to enhance diagnostic sensitivity. Both of these approaches carry the ability to identify malignant cells in samples of limited cellularity and thereby provide enhanced diagnostic sensitivity. SUMMARY: Overall, recognizing the limited sensitivity of diagnostic approaches for tissue diagnosis of cholangiocarcinoma, it would appear prudent to pursue further tissue sampling when there is a strong clinical suspicion of malignancy despite a prior negative/nondiagnostic result. PMID- 19122507 TI - Confocal endomicroscopy. AB - PURPOSE OF REVIEW: Confocal laser endomicroscopy is an emerging field of endoscopic imaging allowing gastroenterologists to obtain in-vivo histology of the gastrointestinal mucosa. The present review will address the more recent advances in confocal laser endomicroscopy and discuss its future. RECENT FINDINGS: Interest in confocal laser endomicroscopy and in-vivo diagnosis of gastrointestinal disorders has increased in the last few years. Confocal laser endomicroscopy can localize intraepithelial neoplasia in chronic ulcerative colitis and enable 'smarter' mucosal biopsy when combined with chromoendoscopy. Confocal laser endomicroscopy may accurately diagnose neoplasia in the esophagus, stomach, colon, and bile duct. Moreover, in-vivo visualization of morphologic epithelial abnormalities, such as intraepithelial bacteria and 'gaps in the gut', may increase our understanding of the possible pathogenic mechanisms of disease. Fluorescent peptide markers that target neoplasia are also being developed as a complementary approach to visualization of in-vivo histopathology. SUMMARY: Confocal endomicroscopy is a developing area of gastrointestinal endoscopy with expanding clinical and research applications. The present review focuses on recent advances in confocal endomicroscopy. PMID- 19122508 TI - Neuroendocrine tumors of the pancreas: endoscopic diagnosis. AB - PURPOSE OF REVIEW: Endoscopic ultrasound (EUS) is a valuable tool in the diagnosis and management of pancreatic neuroendocrine tumors. This review highlights advances over the last year in EUS in the evaluation of pancreatic neuroendocrine tumors. RECENT FINDINGS: We will focus on recent findings regarding the accuracy of EUS, EUS-guided fine needle aspiration (EUS-fine needle aspiration), emerging cytologic markers obtained from fine needle aspiration samples, and the role of EUS screening for patients with multiple endocrine neoplasia type 1 syndrome. Additionally, we will introduce potential therapeutic EUS interventions in the treatment of pancreatic neuroendocrine tumors. SUMMARY: The present review highlights recent advances in the utility of EUS in the clinical management of pancreatic neuroendocrine tumors. Key studies from the last year demonstrate the important role of EUS in the diagnosis, prognosis, and treatment of pancreatic neuroendocrine tumors. PMID- 19122509 TI - Bibliography. Current world literature. PMID- 19122511 TI - Stomach and duodenum. PMID- 19122512 TI - Gastric secretion. AB - PURPOSE OF REVIEW: This review summarizes the past year's literature regarding the regulation and assessment of gastric acid secretion. RECENT FINDINGS: Gastric acid secretion is regulated by biologic agents produced and released by enteroendocrine cells and neurons as well as by exogenously administered substances and infection. Too much acid can lead to gastroesophageal reflux disease, peptic ulcer disease, and stress-related erosion/ulcer disease. Too little acid can interfere with the absorption of certain nutrients, predispose to enteric infection, and interfere with the absorption of some medications. Gastrin, histamine, gastrin-releasing peptide, ghrelin, orexin, and glucocorticoids stimulate whereas leptin, glucagon-like peptide 1, and Helicobacter pylori inhibit acid secretion. Helicobacter pylori inhibits the transcriptional activity of HK-ATPase, the proton pump of the parietal cell. SUMMARY: A better understanding of the pathways and mechanisms regulating gastric acid secretion should lead to improved management of patients with acid-induced disorders as well as those who secrete too little acid. PMID- 19122513 TI - Gastroduodenal mucosal defense. AB - PURPOSE OF REVIEW: The gastrointestinal tract has developed multiple mechanisms of protection from intrinsic and extrinsic sources of injury, including but not limited to drugs, ischemic/reperfusion injuries, and infections such as Helicobacter pylori. We review recent developments in host defense against Helicobacter pylori, duodenal bicarbonate secretion, protection from nonsteroidal anti-inflammatory drug induced gastrointestinal injury, and apoptosis, as well as newer therapies. RECENT FINDINGS: Leptin and survivin confer protection against ethanol and indomethacin induced injury. Mucin-1, a cell surface mucin, is an important barrier to gastrointestinal infection. Prostaglandin E(2), Escherichia coli heat-stable enterotoxin, orexins, and carbonated beverages stimulate duodenal bicarbonate secretion. SUMMARY: Gastroduodenal mucosal defense is a dynamic process, and further insights into these defense mechanisms have and will lead to safer and more effective treatments. PMID- 19122514 TI - Natural orifice translumenal surgery: from scales to symphonies. AB - PURPOSE OF REVIEW: Natural orifice transluminal endoscopic surgery is a new therapeutic procedure that combines surgical and endoscopic skills. Although not currently widely used, it is gaining momentum both in the animal laboratory and in human case reports. RECENT FINDINGS: To date, 15 procedures have been successfully performed on humans worldwide. These included appendectomies (eight), liver biopsies (three), tubal ligation (one), and cholecystectomies (three). Numerous other procedures, using hybrid techniques that combine natural orifice transluminal endoscopic surgery technology with alternative conventional laparoscopic techniques, have been described in human clinical trials. In this review, we will discuss the potential benefits of this procedure over conventional surgical interventions, briefly review the major challenges to the technique (access, spatial orientation, anastomosis, and closure), and point out the technological limitations that severely handicap its potential. SUMMARY: Despite the widespread interest in this technology, there are a lot of shortcomings: limitations in equipment, lack of clinical outcome data, and absence of randomized trials that compare it with alternative conventional surgical interventions. Nevertheless, it is crucial to remember that we have just started the evaluation process for this new exiting technology, and that, clearly, the best is yet to come. PMID- 19122515 TI - Gastritis and gastric atrophy. AB - PURPOSE OF REVIEW: The majority of problems in interpreting gastritis remain Helicobacter related, but their nature has changed. The present review covers gastritis historically through cancer risk staging systems. RECENT FINDINGS: Key points to remember are: Helicobacter is associated with several forms of gastritis; in the present review, I am focusing on the two ends of the disease, 'Helicobacter pylori infection', that starts with antral predominant gastritis but can continue to oxyntic predominant disease with atrophy; the role Helicobacter pylori plays in autoimmune gastritis with pernicious anemia remains unresolved; gastritis staging systems for cancer risk, namely Baylor and Operative Link on Gastritis Assessment, are currently available. SUMMARY: As most gastric carcinomas arise on a background of atrophic gastritis, and the risk increases with the extent of atrophy, an index of atrophy location and extent could be useful in predicting patients at greatest risk for carcinoma. It is now possible to stage patients for cancer risk. Nonetheless, in a field such as gastritis in which many issues remain unresolved, a classification or staging system that is more descriptive will likely prove more useful. PMID- 19122516 TI - Celiac disease. AB - PURPOSE OF REVIEW: Recent advances in the clinical, epidemiological, genetic, and therapeutic aspects of celiac disease have made this condition a superb model of autoimmunity. This review will outline the most significant work that contributed to our current knowledge of the disease. RECENT FINDINGS: Celiac disease is not confined to the Caucasian population as previously believed; rather its prevalence is approximately 1% worldwide. In addition to the HLA genes, many other genes involved in innate and adaptive immunity, intestinal barrier regulation, and autoimmunity have been identified as integral genetic components of the disease. Based on this information and on a better understanding of celiac disease pathogenesis, novel therapies alternative to the gluten-free diet are currently in advanced phase of development. SUMMARY: The outcome of these new findings will most likely have a significant impact in clinical practice, including diagnosis and management of the disease. Furthermore, celiac disease can be used as a unique model to gain more insights on the pathogenesis of autoimmune diseases. PMID- 19122517 TI - Dyspepsia. AB - PURPOSE OF REVIEW: This review critically evaluates the current status of dyspepsia and, in particular, recent advances in epidemiology, pathophysiology and management. The very definition of dyspepsia and of functional dyspepsia, in particular, continues to generate controversy; the Rome III redefinition of functional dyspepsia remains to be proven to be of clinical value. Overlap with gastroesophageal reflux and irritable bowel syndrome further complicate clinical definitions. RECENT FINDINGS: Most studies of pathophysiology continue to focus on gastric sensory and motor functions, though some intriguing early data raise the possibility of an infective or immunological contribution. There have been few, if any, major breakthroughs in treatment; most recent studies address instead the niceties of Helicobacter pylori eradication and acid suppressive strategies. SUMMARY: This continued lack of progress in the area can only lead one to question some very basic concepts in this disorder, such as does functional dyspepsia, as we have come to know it, really exist as a distinct entity? PMID- 19122518 TI - Immunology section. PMID- 19122519 TI - Leaking gut in type 1 diabetes. AB - PURPOSE OF REVIEW: Several studies have indicated that children with type 1 diabetes show altered intestinal immune system and, in particular, increased small intestinal permeability. This review discusses the recent research linking the gut and type 1 diabetes, which may reveal novel pathogenic pathways and new possibilities for disease prevention. RECENT FINDINGS: Recent studies indicate that not only patients with manifest type 1 diabetes show increased small intestinal permeability and high serum levels of zonulin, that is protein controlling epithelial tight junctions, but prediabetic, normoglycemic individuals with beta-cell autoimmunity show signs of leaking gut. Also studies in BioBreeding-rat model of autoimmune diabetes suggest that high permeability of the intestine precedes autoimmune diabetes. The enteropathy characterized by increased intestinal permeability and inflammation seems to be the basis for the development of beta-cell destruction, as for example zonulin agonist, which decreases the gut permeability, prevents the development of diabetes. SUMMARY: The leaking gut syndrome with subclinical inflammation is associated with beta cell autoimmunity and type 1 diabetes. Furthermore, treatment of the leakiness has been reported to modulate development of autoimmune diabetes in animal models suggesting that intestinal environment plays a key role in the destruction of insulin-producing beta-cells in the pancreas. PMID- 19122520 TI - Celiac disease. AB - PURPOSE OF REVIEW: Research in celiac disease is unraveling new findings at a high rate, and major advances seem to occur in all areas such as genetics, environmental factor, pathophysiology, and even prospective therapeutic implications. RECENT FINDINGS: New insight is being gained into the interplay between genetic and environmental factors causing celiac disease. In addition to the known human leukocyte antigen haplotypes, genome-wide studies have now identified additional susceptibility loci and the majority of newly discovered risk regions harbor genes controlling immune pathways. The mechanism of translocation of gliadin peptides across the intestinal barrier has been the subject of much investigation, and there is now evidence that the toxic 33-mer peptide can also be translocated transcellularly. As for the paracellular route, this appears to be enhanced by gliadin's stimulation of zonulin release. The growing role of the innate immunity is being recognized and the increased expression of some Toll-like receptors appears to delineate a new inherent defect in this branch of innate immunity. Finally, new perspectives are opening in the treatment of celiac disease based on new detoxified grains, enzymatic degradation of gluten, and prevention of its crossing the mucosal barrier. SUMMARY: The pace of new knowledge in this 'ancient' disease is very fast, and this review outlines the principal lines of such exciting developments. PMID- 19122521 TI - Oral immunization: an update. AB - PURPOSE OF REVIEW: Oral immunization with vaccines against intestinal infectious diseases has been extensively explored for several decades. Despite the immunologic and economic rationale behind oral immunization, only a few mucosal vaccines are available for the prevention of mucosal infections. Here, we summarize the current status of such vaccines, with a focus on intestinal infectious diseases, describe alternative approaches, and analyze advantages and difficulties encountered with a broad implementation of these vaccines. RECENT FINDINGS: Due to the limited absorption from the intestinal tract and sensitivity to degradation, oral vaccines composed of killed bacteria and viruses or antigens isolated from infectious agents have not been successful. New, live-attenuated bacterial and viral or edible plant-derived vaccines, however, have been recently introduced for this purpose. Furthermore, systemic immunization with vaccines composed of bacterial polysaccharides chemically coupled to suitable protein carriers induces high levels of IgG antibodies, which may provide immunity toward Salmonella typhi, Shigella, and Escherichia coli. SUMMARY: Further improvements in antigen-delivery systems, the development of adjuvants that are safe for mucosal application in humans, use of live-attenuated vaccines and microbial vectors, and production of certain vaccines in plant expression systems are likely to contribute to the broader use of oral vaccines in the future. PMID- 19122522 TI - Symbiotic commensal bacteria direct maturation of the host immune system. AB - PURPOSE OF REVIEW: Although commensal bacteria are known to play an important role in the proper maturation of the immune system of their mammalian hosts, the molecular mechanisms underlying this immunomodulation are poorly characterized. The present review summarizes recent findings in the field and describes new knowledge on the interplay of the innate and adaptive arms of the immune response induced by symbiotic bacterial carbohydrate antigens. RECENT FINDINGS: Commensal bacteria in the intestine not only interact directly with dendritic cells but also engage in cross-talk with epithelial cells. These interactions lead to the induction of tolerogenic antigen-presenting cells in the lamina propria and ultimately to the regulation of functional maturation of effector T cells. Upon recognition of capsular polysaccharide antigens of commensal bacteria by dendritic cells (through toll-like receptor 2), innate immune responses facilitate and act in conjunction with adaptive responses to promote optimal Th1 polarization. In contrast, adaptive immunoglobulin A responses to symbiotic bacteria regulate the magnitude of oxidative innate immune responses in the mucosa as well as bacterial epitope expression in the lumen. SUMMARY: Accumulating evidence is elucidating surface carbohydrate structures of symbiotic bacteria that drive the modulation of the intestinal immune system, resulting in mature, balanced immune responses and oral tolerance. PMID- 19122523 TI - Innate immune signalling at intestinal mucosal surfaces: a fine line between host protection and destruction. AB - PURPOSE OF REVIEW: Emerging evidence underscores that inappropriate innate immune responses driven by commensals contribute to the pathogenesis of chronic inflammatory bowel diseases in genetically susceptible hosts. The present review focuses on defining the recently described mechanistic functions through which the innate immune signalling apparatus shapes mucosal homeostasis of the intestine in health and disease. RECENT FINDINGS: Commensal-induced innate immune signalling actively drives at least six major interdependent functions to control homeostasis in the healthy intestinal mucosa: 1) barrier preservation, 2) inhibition of apoptosis and inflammation, 3) acceleration of wound repair and tissue regeneration, 4) exclusion of harmful pathogens through autophagy and other antimicrobial defenses, while 5) maintaining immune tolerance towards harmless commensals, and 6) linkage to adaptive immunity. Any disturbance of this peaceful and mutually beneficial host-commensal relationship may imbalance innate immune signalling, which predisposes to chronic intestinal inflammation and associated tumourigenesis in inflammatory bowel diseases. SUMMARY: Recent advances have highlighted the complex mechanistics and functional diversity of innate immunity that paradoxically mediate both protective and destructive responses in the intestinal mucosa. Related signalling targets may offer novel therapeutic approaches in the treatment of inflammatory bowel diseases and inflammation-related cancer. PMID- 19122524 TI - Clinical and molecular aspects of autoimmune enteropathy and immune dysregulation, polyendocrinopathy autoimmune enteropathy X-linked syndrome. AB - PURPOSE OF REVIEW: Autoimmune enteropathy (AIE) is a distinct cause of severe and persistent inflammatory diarrhea in children. Recent research data allowed us to gain a first insight in the pathogenesis of AIE. On the basis of this data, we will discuss new aspects of AIE emphasizing new diagnostic and therapeutic possibilities. RECENT FINDINGS: With the discovery of disease-causing mutations in the FOXP3 gene in patients with AIE, a dramatic advance in the understanding of AIE was made. Subsequent studies indicated that FOXP3 is a key transcription factor indispensable for regulatory functions of T cells pointing to a critical role of regulatory T-cell homeostasis in the development of AIE. Abnormal FOXP3 expression results in defective regulatory functions of T cells, which in turn cause a systemic T-cell-mediated autoaggressive disorder, now called immune dysregulation, polyendocrinopathy autoimmune enteropathy X-linked syndrome. Upon systematic review, we describe different phenotypes of immune dysregulation polyendocrinopathy autoimmune enteropathy X-linked syndrome, as well as immune dysregulation polyendocrinopathy autoimmune enteropathy X-linked-like forms of AIE, which are FOXP3 independent. No genotype-phenotype correlation could be established so far. SUMMARY: On the basis of the profound immune dysregulation in AIE, new, most often T-cell-oriented treatment strategies were developed. The recent molecular advances in the understanding of AIE give a clear rational for the use of immunosuppression (combining steroids and tacrolimus or rapamycine) to stabilize AIE patients or to perform bone marrow transplantation in those who do not respond to immunomodulation. PMID- 19122525 TI - Bibliography. Current world literature. PMID- 19122527 TI - Adolescent suicide. AB - PURPOSE OF REVIEW: The present review summarizes the updated literature on adolescent suicide. RECENT FINDINGS: Reductions in youth suicide rates are probably related to use of selective serotonin reuptake inhibitors since the mid 1990s as well as restrictions in means and enhanced pesticide control. The serotonin theory of suicide has received more empirical support. Familial transmission of suicidal behavior may be mediated by transmission of impulsive aggression from parent to child and early detection of precursors of suicidal behavior can help identify families at high risk of having a suicidal child. A newly investigated social risk factor of bullying adolescents and the novel psychological construct of autobiographical memory all help to advance our understanding and treatment of suicidal youths. Much effort is needed in establishing more solid empirical evidence for suicide prevention programs and treatment, while assessment tools are still in desperate need of further development. SUMMARY: Suicidal behavior remains an important clinical problem and a major cause of death in youth. There are key issues that need to be solved for better prediction of suicidality, prevention and treatment of youth suicide. PMID- 19122528 TI - Augmentation strategies for treatment-resistant depression. AB - PURPOSE OF REVIEW: The majority of patients with depression fail to remit on one or more antidepressant trials. These patients have treatment-resistant depression (TRD) with high relapsing rates. Augmentation pharmacotherapy refers to the addition of drugs that are not standard antidepressants in order to enhance the effect of a classical antidepressant drug. This review highlights the current status and future research directions of augmentation treatments for TRD with a special focus on research data published within the past year. RECENT FINDINGS: Atypical antipsychotics, stimulants, pindolol, lithium, lamotrigine and mecamylamine were tested for efficacy in clinical trials. Most of the trials were not controlled or had limited sample size. Recent data now support the use of some atypical antipsychotics to augment depression resistant to the newer, more selective, antidepressants. SUMMARY: Lithium and triiodothyronin (T3) augmentation of tricyclic agents remains the best studied strategy. Data converge to demonstrate the efficacy of some atypical antipsychotics as augmenting agents to selective serotonin reuptake inhibitors. Further adequately powered controlled trials on augmentation pharmacotherapy of TRD are necessary. PMID- 19122529 TI - Managing unipolar depression in pregnancy. AB - PURPOSE OF REVIEW: The management of depression in pregnancy is complex, as it is based on balancing the risks with the benefits of treatment versus no treatment for both the mother and the fetus. The current literature in the field of reproductive psychiatry is difficult to navigate and at times contradictory. This article aims to review both nonpharmacological and pharmacological modalities in the treatment of perinatal depression. A literature review using PubMed and MEDLINE databases was used to collect literature from the past 2 years; however, given the relatively small amount of research in reproductive psychiatry, several salient articles from the past 5 years have also been included in this review. RECENT FINDINGS: Recent US Food and Drug Administration and Health Canada warnings regarding poor neonatal adaptation and adverse perinatal outcomes associated with antidepressant use in pregnancy have changed patterns of practice for prescribing physicians. Many physicians are now left with a sense of indecisiveness regarding the safety of treating their depressed, pregnant patients. Similarly, these warnings have changed patients' attitudes and their willingness to consider pharmacological treatment for depression. Although these warnings demand attention and careful consideration, research has also shown that exposure to mental illness in pregnancy has deleterious short-term and long-term effects for the exposed mother and fetus. SUMMARY: The field of reproductive psychiatry is rapidly evolving. Clinicians need to keep abreast of changes in the management of depression during pregnancy. Ongoing research in this field is important so that the most up-to-date recommendations may be provided to pregnant women. PMID- 19122530 TI - Cognitive abnormalities in posttraumatic stress disorder. AB - PURPOSE OF REVIEW: The current review aims to describe and evaluate research on cognitive difficulties associated with posttraumatic stress disorder (PTSD) published between January 2007 and June 2008. RECENT FINDINGS: The reviewed studies provide additional evidence that negative appraisals and decrements on verbal and autobiographical memory tasks are exhibited by individuals with PTSD relative to controls and may represent preexisting risk factors for PTSD rather than a result or concomitant of PTSD symptoms. In addition, the reviewed findings provide further evidence for source monitoring difficulties and attentional biases toward trauma-relevant information in PTSD. Recent research also provides evidence that, although memories of traumatic events differ in PTSD relative to other types of memories and memories of depressed individuals, they are likely on the same continuum as nontraumatic memories rather than qualitatively different. SUMMARY: Individuals with PTSD experience cognitive alterations ranging from impairments in overall memory functioning to difficulties specific to trauma related cues. These cognitive difficulties appear to be importantly related to the development and/or maintenance of the disorder. At this point, it is unclear whether common mechanisms may account for these diverse cognitive difficulties and whether cognitive impairments are attributable to comorbid depression. PMID- 19122531 TI - Neurosurgical treatment of mood disorders: traditional psychosurgery and the advent of deep brain stimulation. AB - PURPOSE OF REVIEW: From its peak in the 1940s and 1950s, psychosurgery (or, neurosurgery for psychiatric disorders) has had a gradual decline, with only a few centers around the world continuing with the procedure into the 1980s and 1990s. With recent developments in brain stimulation techniques, the continuing relevance of psychosurgery in the treatment of psychiatric disorders is worthy of examination. RECENT FINDINGS: A review of databases (PubMed, Medline, Current Contents and Embase) suggests that psychosurgery in the form of stereotactic focal ablation is still practiced in a few centers, although the number has decreased further from the 1990s. Procedures have not changed substantively, although modern imaging and stereotaxy have made them more precise. No good predictors of treatment response have been identified. There is a major shift in interest to deep brain stimulation (DBS) instead of ablative surgery. Studies of DBS in resistant depression and obsessive-compulsive disorder have been few and have involved small numbers, but this field is growing rapidly. SUMMARY: Although ablative psychosurgery using stereotactic procedures continues to be used to a small extent, psychiatrists remain ambivalent about this procedure. The baton of psychosurgery, however, appears to have been passed on to DBS, but more data are needed on technical details and outcomes before the possible therapeutic role of DBS can be established. PMID- 19122532 TI - Inflammatory markers in depression. AB - PURPOSE OF REVIEW: To provide a critical update of the literature linking depression and inflammation, together with possible underlying mechanisms and longer term risk of cardiovascular disease. RECENT FINDINGS: The current literature lends further support to the view that major depression is associated with a proinflammatory response, as indexed by elevation in C-reactive protein and cytokines such as interleukin 6 and tumour necrosis factor-alpha. Antidepressants suppress the inflammatory response, whereas electroconvulsive therapy acutely increases proinflammatory cytokine levels. Most, though not all, studies support a link between depression, inflammation and cardiovascular events. SUMMARY: Depression is an inflammatory state that may increase the risk of cardiac disease. Whether or not the immune system is an appropriate target for antidepressant development has yet to be established. PMID- 19122533 TI - Collaborative care for mood disorders. AB - PURPOSE OF REVIEW: Psychiatric and health services research literature was reviewed to identify recent studies regarding effectiveness, cost-effectiveness, and generalizability of collaborative care programs to improve treatment of mood disorders. RECENT FINDINGS: Meta-analyses of previous randomized trials as well as recently conducted trials strongly support the effectiveness of collaborative care programs for depression across a broad range of primary care settings. Over time, the direct costs of providing more effective treatment for depression appear to be balanced by decreases in the use of general medical services, especially for patients with comorbid chronic medical illness. Given that the added costs of collaborative care are incurred early and economic benefits appear slowly, successful implementation of collaborative care programs will depend on adequate funding and availability of dedicated staff. Growing evidence supports the effectiveness of collaborative care programs to improve management of bipolar disorder in mental health specialty settings. SUMMARY: Given the strong evidence for the effectiveness, cost-effectiveness, and generalizability of collaborative care programs for depression in primary care, attention should turn to dissemination and implementation. Additional research is needed regarding effectiveness of collaborative care programs for more severe psychiatric disorders. PMID- 19122534 TI - Rating scales in bipolar disorder. AB - PURPOSE OF REVIEW: Bipolar disorder is a complex clinical entity that presents with pleomorphic signs and symptoms varying from depression to hypomania, full blown mania, mixed states, and psychosis. Therefore, the assessment of patients with bipolar disorder poses a difficult challenge to clinicians, who may find themselves puzzled when having to choose among the many instruments available. This paper aims at briefly reviewing these instruments. RECENT FINDINGS: Observer rated and self-completed instruments for the assessment of depressive, manic, and psychotic symptoms in patients with bipolar disorder are described with emphasis on topics of increasing interest in recent times, such as the assessment of bipolar depression, self-rating of manic symptoms, daily mood monitoring, screening, and early recognition of bipolar disorder. SUMMARY: The use of standardized assessment instruments has many advantages as it enables clinicians to be more systematic and it allows greater precision, increased sensitivity to change, and opportunity to identify longitudinal patterns. Knowledge of available options for the assessment of patients with bipolar disorder may help clinicians and researchers to choose the most appropriate instruments for their specific needs. PMID- 19122535 TI - It is too early for adjusting the adjustment disorder category. AB - PURPOSE OF REVIEW: Despite the relative frequency with which the diagnosis of adjustment disorder is made, there is a very limited research literature in regard to its cause, epidemiology and treatment. This review summarizes recent papers and findings in relation to this diagnostic category. RECENT FINDINGS: The conceptual underpinnings of the diagnosis adjustment disorder is the subject of ongoing debate as is its differentiation from other psychiatric disorders such as depressive disorders. One group has proposed and tested a diagnostic model of adjustment disorder as a stress-response syndrome, related to posttraumatic stress disorder, which is a distinct psychopathological entity rather than a diagnosis of exclusion. Initial neuroimaging work in adjustment disorder indicates that this may be a helpful approach for increasing understanding of the biology of the disorder. Recent studies support the use of brief psychotherapy in the treatment of adjustment disorder. SUMMARY: Although the studies and articles described in this review raise some interesting questions in relation to the diagnosis, cause, epidemiology and treatment of adjustment disorder, much more research is needed before we can draw some firm conclusions about the need for the redefinition and reclassification of this diagnostic category. PMID- 19122536 TI - Current empirical status of acceptance and commitment therapy. AB - PURPOSE OF REVIEW: Acceptance and commitment therapy (ACT) is one of the several psychotherapies that have been described as 'third wave' cognitive behavioral therapies (CBTs). The present editorial review examines the current status of ACT with a focus on previous reviews, a meta-analysis and new studies that have been published between January 2006 and August 2008. RECENT FINDINGS: Recent studies on ACT suggest that ACT may be effective for a variety of disorders, including several anxiety disorders, depression, pain, trichotillomania, psychotic disorder, drug abuse and the management of epilepsy and diabetes. SUMMARY: The available evidence suggests that ACT works through different processes than active treatment comparisons, including traditional CBT. Although currently available data are promising, there is, however, a need for more well controlled studies to verify whether ACT is generally as or more effective than other active treatments across the range of problems examined. PMID- 19122537 TI - Quality of life and neurotic disorders in general healthcare. AB - PURPOSE OF REVIEW: To review recent findings on the relationship between neurotic disorders (specifically depression, anxiety and somatization) and function and quality of life (QoL) in patients with complex clinical presentations, and to consider the clinical and public health implications. RECENT FINDINGS: The high prevalence of comorbid depression and anxiety among patients with general medical disorders is well known. The impact of the comorbidity on function in these patients is documented extensively. The study of their QoL is an emerging area of work, though its interpretation is controversial. The impact of somatization on function and QoL is less well studied. Recent findings emphasize the value of a common measure for QoL across health conditions. SUMMARY: Depression, anxiety and somatization are a significant burden in the general population and failure to recognize and manage these problems in general healthcare contributes to delayed recovery. A holistic approach to symptom, disability and QoL assessment, done in awareness of possible measurement redundancy, is likely to assist better recognition and management of these conditions. Further research is needed on the effects on QoL of depression and anxiety in patients with general medical disorders and early detection and treatment of somatization. PMID- 19122538 TI - Treatment of pathological gambling. AB - PURPOSE OF REVIEW: This paper highlights the development of pharmacological and nonpharmacological treatments for pathological gambling and is based on a review of the literature published in the past 12 months. RECENT FINDINGS: The efficacy of naltrexone treatment for pathological gambling has been replicated in a double blind, placebo-controlled, confirmatory study. For mood stabilizers, whereas carbamazepine and topiramate continued to produce positive results, olanzapine failed to show superior outcomes compared with placebo control. Two new pharmacological agents for pathological gambling, N-acetyl cysteine and modafinil, produced significant improvement for pathological gamblers. Several studies examined the outcomes of nonpharmacological treatments. Recent studies showed that cognitive-behavioral therapy failed to produce superior outcomes compared with other less costly methods such as brief interventions. Two new nonpharmacological treatment methods have been reported, including the use of videoconferencing in delivering ongoing supervisions after exposure therapy and the congruence couple therapy, which aims to heal the person as a system whole. SUMMARY: Recent treatment outcomes studies address not only the effectiveness, but also the efficacy of different treatment approaches. Results of two meta analysis studies showed that nonpharmacological treatments have a larger overall effect size than pharmacological treatments; however, owing to the diversity in study designs, it is unclear whether nonpharmacological treatments are more effective than pharmacological treatments at this point. PMID- 19122539 TI - Long-term outcome of anxiety disorders: a review of double-blind studies. AB - PURPOSE OF REVIEW: Although anxiety disorders are acknowledged as chronic, the issue of the pharmacological treatment duration remains unsettled. This review focuses on the long-term outcome of patients with anxiety disorders as demonstrated by randomized controlled trials. RECENT FINDINGS: Results from long term randomized controlled trials of antidepressants in anxiety disorders indicate that maintenance treatment significantly reduces the odds of relapse, whatever the anxiety disorder is. This result appears to be similar to what is reported in long-term studies in depressive disorders. In addition, regarding the natural course of depressive disorders, acknowledged as mostly recurrent, some patients may require very long-term treatment, that is, more than 2 years. Moreover, naturalistic studies in anxiety disorders indicate that the relapse risk after discontinuation is not associated with the treatment duration. Finally, there is no predictor to identify those patients who require long-term pharmacotherapy for anxiety disorders. SUMMARY: In light of this review, other long-term studies in anxiety disorders have to be undertaken to identify predictors of relapse after treatment discontinuation. As it is now acknowledged for depressive disorders, some patients may require very long-term pharmacological treatment for anxiety disorders. PMID- 19122540 TI - Current use of benzodiazepines in anxiety disorders. AB - PURPOSE OF REVIEW: The aim of this study is to provide a review of articles published between July 2007 and August 2008 on the current use and rationale of benzodiazepines in anxiety disorders. RECENT FINDINGS: Recent review articles confirm selective serotonin reuptake inhibitors as first-choice drugs for treating anxiety disorders, alongside newer agents such as pregabalin or serotonin-norepinephrine reuptake inhibitors, and combined with cognitive behavioural therapy. Benzodiazepines are still widely used by clinicians for these disorders, as shown by recent surveys, even though their anxiolytic effectiveness is questioned. Newer agents are in development and may in the future resolve the therapeutic dilemma. SUMMARY: Despite current guidelines, benzodiazepines are still considered by many clinicians to remain good treatment options, in both the acute and the chronic phase of the treatment of anxiety disorders, partially because of their rapid onset of action and their efficacy with a favourable side effect profile, and also because of the sometimes only incomplete therapeutic response and the emergence of side effects of alternative medications. Having experienced good initial symptom relief with benzodiazepine treatment, patients may also be reluctant to taper it down. Clinicians should, however, bear in mind the frequent physiological dependence associated with these substances, and suggest both pharmacological and psychological treatment alternatives before opting for a long-term benzodiazepine treatment, which may remain necessary in certain clinical conditions. PMID- 19122541 TI - Current status of brain imaging in anxiety disorders. AB - PURPOSE OF REVIEW: This study reviews the most recent literature about brain imaging research in anxiety disorders. There is a growing body of evidence that neuroimaging of anxiety disorders contributes to a better understanding of the neurobiology of these disorders, by identifying cerebral modifications occurring previously or subsequent to symptoms of anxiety. A systematic search of the literature (January 1978-July 2008) was performed in MEDLINE using the keywords brain imaging, anxiety disorders, posttraumatic stress disorder, obsessive compulsive disorder, generalized anxiety disorder, specific phobia, social anxiety disorder, panic disorders and agoraphobia. References cited in all trials were searched iteratively to identify missing studies. Our review focused only on the last year's findings. RECENT FINDINGS: There is a consensus on the crucial role of the amygdala, anterior cingulate cortex and insula in the pathophysiology of anxiety disorders. SUMMARY: Brain imaging research in anxiety disorders has become increasingly important, especially in the last decade, because of the opportunity to validate neurobiological hypotheses for anxiety disorders. Thus, neuroimaging data raise the question of the neurobiological cause of anxiety disorders, opening up new reflections not only on pharmacological treatments but also on the nosology of the anxiety disorders. PMID- 19122542 TI - DSM axis II: personality disorders or adaptation disorders? AB - PURPOSE OF REVIEW: After the initial enthusiasm, the study of personality disorder seems to be at a crossroad, without clear direction. This is mainly due to overlapping categorical diagnostic criteria of personality disorders. Study samples based on these criteria are inadequate and their results questionable. RECENT FINDINGS: The literature is unanimously advocating a dimensional concept of personality disorders. Four dimensions are consistently reported to underlie personality disorder symptoms. We put forward an argument that personality disorders are disorders of adaptation, not of personality per se, as extreme personality traits are not ipso facto dysfunctional. Available methods to assess maladaptation are reviewed. SUMMARY: The diagnosis 'personality disorder' should be replaced by the diagnosis 'adaptation disorders'. This reflects the real nature of the disorder more accurately, and is likely to reduce the stigmatizing component of the personality disorder diagnosis as it places emphasis on positive efforts to improve adaptation. The suggested revisions of the personality disorder diagnosis and dimensional approach to these disorders are likely to advance treatment and research - we discuss these aspects in some detail. PMID- 19122543 TI - Bibliography. Current world literature. PMID- 19122544 TI - Ventilatory management during routine general anaesthesia. AB - Intraoperative hypoxaemia and postoperative respiratory complications remain the challenges of modern anaesthetic practice. Anaesthesia causes both depression of respiratory centres and profound changes of respiratory mechanics. Most anaesthetized patients consequently require mechanical ventilation and supplemental oxygen. Recent data suggest that intraoperative respiratory management of a patient can affect postoperative outcome. In this review, we briefly describe the mechanisms responsible for the impairment of intraoperative gas exchange and provide guidelines to prevent or manage hypoxaemia. Moreover, we discuss several aspects of mechanical ventilation that can be employed to improve patients' outcome. PMID- 19122545 TI - Low-frequency changes in finger volume in patients after surgery, related to respiration and venous pressure. AB - BACKGROUND AND OBJECTIVE: In patients after surgery, we observed large-amplitude low-frequency changes in digital plethysmograph measurements when DC coupling of the signal was used. We set out to assess factors that might contribute to these events and in particular to test the possibility that low-frequency signals could be used to assess respiratory disturbances. METHODS: We recorded values in 23 patients who had undergone gynaecological surgery. We measured nasal flow, abdominal pressure (by urinary catheter), venous pressure in the hand, and DC coupled optical transmission plethysmography. Signals were replayed and analysed to assess the incidence of specific patterns of events. RESULTS: Most patients received morphine for postoperative analgesia. Respiratory irregularity and expiratory muscle action were very frequent. Increases in abdominal pressure during expiration caused increases in venous pressure and pulsation. In 12 out of 23 patients, a characteristic response consistent with vasoconstriction was noted after increases in breath size, and, in seven patients, very-low-frequency (0.2 0.7 Hz) oscillations of finger volume were present that appeared unrelated to respiratory events. Patients who did not receive morphine had very different plethysmograph patterns, with significantly smaller pulse amplitude. CONCLUSION: Low-frequency changes in finger volume can be simply obtained and provide considerable information about peripheral circulatory dynamics. Diverse patterns can be recognized, but the range of responses suggests that current techniques cannot be used alone to assess cardiorespiratory status. However, a combination of plethysmography with respiratory measurements shows characteristic events. PMID- 19122546 TI - Retention and transferability of team resource management skills in anaesthetic emergencies: the long-term impact of a high-fidelity simulation-based course. AB - BACKGROUND AND OBJECTIVE: In 2005, we developed and implemented the Emergency Anaesthetic Simulated Experience course at the Cheshire and Merseyside Simulation Centre.Emergency Anaesthetic Simulated Experience aims to teach clinical and team resource management skills to junior trainees in anaesthesia. Here we present 'proof-of-concept' in terms of long-term retention and transferability of acquired skills into subsequent clinical practice. METHODS: An electronic questionnaire sent to 73 trainees, 9-20 months after the course, invited open ended responses, addressing four areas; namely, real-life encounters with the same scenario as on the course, approach to real-life anaesthetic emergencies in general, approach to real-life routine anaesthesia and need to attend similar courses in the future, with their underlying reasons. RESULTS: Qualitative analysis of the descriptive responses showed that the lessons learnt in the context of simulated emergencies were applied by candidates themselves to real life emergencies in general and to routine practice. CONCLUSION: Team resource management skills learnt in a single educational intervention, based on simulated anaesthetic emergencies, are retained over the long term, translated into clinical practice and are transferable across the breadth of clinical activities. PMID- 19122547 TI - Compatibility and stability of binary mixtures of acetaminophen, nefopam, ketoprofen and ketamine in infusion solutions. AB - BACKGROUND AND OBJECTIVE: Administering various combinations of acetaminophen, ketoprofen, nefopam and ketamine, though sometimes discussed, is expected to provide superior pain relief and reduce opioid analgesic-related side effects. However, some studies have indicated that multimodal analgesia has limited efficacy. We studied the stability of various binary combinations of these four drugs. PATIENTS AND METHODS: The drugs were studied at 25 degrees C. Binary mixtures of acetaminophen, ketoprofen, nefopam and ketamine were produced. Each drug concentration was assessed using a specific high-performance liquid chromatographic technique. Measurements were carried out at T0, +1, +2, +4, +6 and +24 h. A 5% loss of initial concentration was considered to be significant. The changes with time of the concentrations were analysed using linear regression analysis. A P value of less than 0.05 was significant. RESULTS: The four drugs tested in the binary mixtures were stable, showing neither loss of concentration nor degradation products (P > or = 0.05). CONCLUSION: Physicochemical negative interaction is not likely to account for the limited clinical efficacy sometimes reported with binary combinations of these drugs. Coadministration of binary mixtures of acetaminophen, nefopam, ketoprofen and ketamine from the same bottle or infusion bag using the same venous line is demonstrated to be feasible. PMID- 19122548 TI - Continuous infusion of butorphanol combined with intravenous morphine patient controlled analgesia after total abdominal hysterectomy: a randomized, double blind controlled trial. AB - BACKGROUND AND OBJECTIVE: Postoperative pain therapy is still a problem for clinicians. Continuous basal infusion of drugs combined with standard patient controlled analgesia (PCA) is considered to be an effective means of postoperative acute pain management. This study was designed to investigate the analgesic efficacy, morphine-sparing effects and side effects of butorphanol delivered as a continuous infusion adjunct to intravenous morphine PCA after abdominal hysterectomy. METHODS: One hundred and eighty-six ASA physical status I II patients, undergoing total abdominal hysterectomy, were allocated to this randomized double-blind controlled study and assigned to one of two groups. In the butorphanol (n = 96) group, patients received an intravenous loading dose of 10 microg kg(-1) butorphanol followed by infusion of 2 microg kg(-1) h(-1) butorphanol combined with intravenous PCA set at a bolus of 0.02 mg kg(-1) morphine after surgery. The control group (n = 90) received a physiological saline infusion combined with the same morphine PCA. Pain intensity on movement and at rest, sedation, satisfaction with analgesia, morphine consumption and side effects were recorded. RESULTS: A total of 164 patients completed the study. The butorphanol group had analgesia superior to the physiological saline control (P < 0.001). The butorphanol infusion group produced higher sedation ratings (P < 0.001) and better satisfaction (P < 0.05) and a lower incidence of side effects (P < 0.001) with the exception of sweating and dry mouth (P < 0.05) than the physiological saline group. The butorphanol group consumed less morphine over 48 h, 24.6 mg (95% confidence interval, 18.7-46.6), than the physiological saline group, 58.5 mg (95% confidence interval, 41.5-79.2; P = 0.006). There were no differences between urinary catheterization of more than 24 h, first time out of bed and time to discharge to home. CONCLUSION: Basal infusion of butorphanol combined with intravenous morphine PCA in patients undergoing abdominal hysterectomy shows effective analgesia with sedation and fewer side effects. PMID- 19122549 TI - Saving sevoflurane and hastening emergence from anaesthesia using an anaesthetic conserving device. AB - BACKGROUND AND OBJECTIVE: This study compared an anaesthetic-conserving device (AnaConDa) and a conventional vaporizer in terms of sevoflurane consumption and emergence from anaesthesia using a total gas flow of 4 l min(-1), with a rebreathing circuit. PATIENTS AND METHODS: Twenty-four patients for gastrectomy, aged 30-70 years, were divided into AnaConDa and control groups. Anaesthesia was induced with midazolam, propofol, fentanyl and vecuronium. The total gas flow was fixed at 4 l min(-1) (nitrous oxide 2 l min(-1) and oxygen 2 l min(-1)). Sevoflurane administration was started at 0.5% (vaporizer setting) in the control group and 25 ml h(-1) in the AnaConDa group, then the end-tidal sevoflurane concentration was kept between 0.3 and 0.5% in both groups. Analgesia was obtained with intermittent epidural administration of mepivacaine. The time to first detection of end-tidal sevoflurane, sevoflurane consumption and emergence time were compared between the two groups. RESULTS: The AnaConDa group showed a significantly longer time to first detection of end-tidal sevoflurane (211 +/- 75 vs. 40 +/- 18 s), smaller sevoflurane consumption (12 +/- 3 vs. 42 +/- 9 ml), and shorter emergence time (12 +/- 2 vs. 16 +/- 1 min) than the control group. CONCLUSION: The AnaConDa could decrease sevoflurane consumption and hasten emergence from anaesthesia, but increasing sevoflurane concentration with AnaConDa at the start of anaesthesia might take longer than that with a conventional vaporizer. PMID- 19122550 TI - Landiolol, a new ultra-short-acting beta1-blocker, reduces anaesthetic requirement during sevoflurane/N(2)O/fentanyl anaesthesia in surgical patients. AB - BACKGROUND AND OBJECTIVE: It is known that esmolol, a short-acting beta1-blocker, reduces anaesthetic requirement. In this study, we evaluated whether a low dose of landiolol, a new ultra-short-acting beta1-blocker, can reduce the sevoflurane requirement. METHODS: Twenty-five patients undergoing hip surgery were randomly divided into two groups. Group A (n = 13) received landiolol (bolus injection of 0.031 mg.kg(-1) and continuous infusion at a rate of 0.01 mg.kg(-1).min(-1)). Group B (n = 12) received physiological saline. Landiolol and physiological saline were started before the induction of anaesthesia and continued until the end of anaesthesia. Anaesthesia was maintained with sevoflurane, 60% N(2)O and fentanyl. Sevoflurane concentration was controlled to keep the bispectral index at approximately 50. The end-tidal sevoflurane concentration and haemodynamics were measured during anaesthesia. RESULTS: The average end-tidal sevoflurane concentration in group A was significantly lower than that in group B (1.2 +/- 0.30 vs. 1.8 +/- 0.3%, P < 0.01). Maximum values of systolic arterial pressure showed no difference between the groups, whereas the maximum value of heart rate in group A was significantly less than that in group B (61 +/- 10 vs. 76 +/- 14 beats min(-1), P < 0.05). CONCLUSION: The results suggest that a low dose of landiolol significantly reduces the intraoperative sevoflurane requirement during sevoflurane/N(2)O/fentanyl anaesthesia in patients undergoing hip surgery. PMID- 19122551 TI - Delineation of distal ulnar nerve anatomy using ultrasound in volunteers to identify an optimum approach for neural blockade. AB - BACKGROUND AND OBJECTIVE: Ultrasound can provide novel approaches to neural blockade independent of surface landmarks. We elucidated the sonoanatomy of the ulnar nerve in the forearm of healthy volunteers in order to identify an optimum site for neural blockade. METHODS: One hundred forearms were scanned; the shape, depth from skin and cross-sectional area of the nerve were noted at the elbow, forearm and wrist. RESULTS: The nerve was visualized in all volunteers and had a maximum depth of 18.9 mm from the skin. The mean distance between the nerve and artery, 2 cm proximal to the point where the two structures met in the forearm, was 8.5 mm (95% confidence interval 8.1-8.9 mm). This was approximately at the junction between the proximal 2/5 and distal 3/5 of the forearm. CONCLUSION: Our study demonstrates that ultrasound can be utilized to identify the ulnar nerve and artery in the forearm. This implies that traditional landmarks will not be required prior to neural blockade. We have suggested a point for blockade of the nerve to reduce risk of vascular puncture. PMID- 19122552 TI - Randomised controlled trial of combined spinal epidural vs. spinal anaesthesia for elective caesarean section: vasopressor requirements and cardiovascular changes. AB - BACKGROUND AND OBJECTIVES: Combined spinal and epidural anaesthesia (CSEA) has previously been shown to result in a higher sensory block than equivalent single shot spinal anaesthesia (SSSA). In nonpregnant patients, hypotension was also more pronounced in the CSEA group. The aim of this randomized trial was to compare the haemodynamic stabilities of CSEA and SSSA during elective caesarean section when the same dose of anaesthetic was administered. This was studied directly by measuring the noninvasive arterial blood pressure (BP) and indirectly by the amount of ephedrine required to maintain baseline BP. Systemic vascular resistance index (SVRI) and cardiac index (CI) were also measured using thoracic impedance cardiography. METHODS: Seventy women received hyperbaric bupivacaine (12.5 mg) and diamorphine (0.3 mg) intrathecally via either CSEA or SSSA. Noninvasive arterial BP, CI and SVRI were measured every 2 min. The total ephedrine used was calculated. RESULTS: There were no significant differences between the groups in ephedrine requirements (P = 0.38), intraoperative mean arterial pressure (P = 0.77), CI (P = 0.17) or SVRI (P = 0.10). CONCLUSION: CSEA placement appears to offer no haemodynamic benefits compared with SSSA when the same dose of local anaesthetic is administered. PMID- 19122554 TI - Propofol suppresses activation of the nociception specific neuron in the parafascicular nucleus of the thalamus evoked by coronary artery occlusion in rats. AB - BACKGROUND AND OBJECTIVE: The neural response to acute myocardial ischaemia was reported as increases in neuronal activity of spinal projecting neurons and the neurons in the parafascicular nucleus (Pf) of the thalamus. The aim of this study was to investigate electrophysiologically the response of the nociception specific neuron (NSN) in the Pf to coronary artery occlusion (CAO) and a potential antinociceptive effect of propofol on the response in rats. METHODS: The left anterior descending branch of the coronary artery was instrumented with a snare occluder in anaesthetized Sprague-Dawley rats. The unit discharges of the NSN were recorded using a single-barrel glass microelectrode and analysed to examine the response to CAO. Once the effect of CAO on the discharge was detected, the antinociceptive effect of propofol was examined. RESULTS: The NSN (30/46) showed a significant increase in the discharge rate following CAO. The increased discharge was suppressed immediately after intravenous injection of propofol at doses of 0.2 and 2 mgkg(-1). The inhibitory effect reached the maximum at 7 min (59.78 +/- 11.17% of suppression, P < 0.05) and 5 min (95.21 +/- 4.79% of suppression, P < 0.05) following administration of 0.2 and 2 mgkg(-1) of propofol, respectively. The effect of propofol was dose dependent and reversible. The maximum dose of propofol used in this study did not produce an anaesthetic effect and did not affect the spontaneous activity of the NSN. CONCLUSION: The findings of this study suggest that propofol plays a role in modulation of the nociception associated with acute myocardial ischaemia, displaying an antinociceptive effect on cardiac visceral pain evoked by acute myocardial ischaemia. PMID- 19122553 TI - Effects of spinal anaesthesia versus epidural anaesthesia for caesarean section on postoperative analgesic consumption and postoperative pain. AB - BACKGROUND AND OBJECTIVE: Regional anaesthesia is commonly used for elective caesarean section. The aim of this study was to investigate whether there is a positive effect of either spinal or epidural anaesthesia on postoperative analgesic requirements and pain relief. METHODS: The analgesic effect of either spinal or epidural induction of perispinal anaesthesia have been compared in 132 women (ASA I or II) scheduled for elective caesarean section, all having epidural catheterization for perioperative anaesthesia and postoperative analgesia. The patients were randomized into two groups. To achieve a sensory block height to the level of the sixth thoracic dermatome, the parturients received isobaric bupivacaine 0.5% and 5 microg sufentanil intrathecally or ropivacaine 0.75% and 10 microg sufentanil epidurally. For postoperative analgesia, all patients used patient-controlled epidural analgesia at identical settings [bolus of ropivacaine 0.133% (11-15 mg according to patient's height), lock-out time 1 h]. Intraoperative and postoperative pain was recorded using a visual analogue pain score as well as analgesic requirements over the first 24 h after surgery. RESULTS: One hundred and twenty-five patients completed the study. There were no differences in patient-controlled epidural analgesic requirements between groups. During surgery, the pain score on a visual analogue scale was more intense with epidural anaesthesia than with spinal anaesthesia (P < 0.05). For the whole 24 h observation period, the area under the curve for pain was lower with spinal anaesthesia (P < 0.0005). At almost all postoperative time points, visual analogue scale scores at rest and during mobilization were lower with spinal anaesthesia (P < 0.05), which was accompanied by less motor blockade and lower frequency of adverse effects. More patients with epidural anaesthesia received supplemental analgesic medication. CONCLUSION: In parturients undergoing elective caesarean section, postoperative use of epidural ropivacaine via patient controlled epidural analgesia is similar after spinal and epidural anaesthesia. Spinal anaesthesia is, however, accompanied with less postoperative pain, use of additional analgesics and side-effects. PMID- 19122555 TI - Influence of ventilatory settings on static and functional haemodynamic parameters during experimental hypovolaemia. AB - BACKGROUND AND OBJECTIVE: The influence of ventilatory settings on static and functional haemodynamic parameters during mechanical ventilation is not completely known. The purpose of this study was to evaluate the effect of positive end-expiratory pressure, tidal volume and inspiratory to expiratory time ratio variations on haemodynamic parameters during haemorrhage and after transfusion of shed blood. METHODS: Ten anaesthetized pigs were instrumented and mechanically ventilated with a tidal volume of 8 mlkg(-1), a positive end expiratory pressure of 5 cmH(2)O and an inspiratory to expiratory ratio of 1:2. Then, they were submitted in a random order to different ventilatory settings (tidal volume 16 mlkg(-1), positive end-expiratory pressure 15 cmH(2)O or inspiratory to expiratory time ratio 2:1). Functional and static haemodynamic parameters (central venous pressure, pulmonary artery occlusion pressure, right ventricular end-diastolic volume and pulse pressure variation) were evaluated at baseline, during hypovolaemia (withdrawal of 20% of estimated blood volume) and after an infusion of withdrawn blood (posttransfusion). RESULTS: During baseline, a positive end-expiratory pressure of 15 cmH(2)O significantly increased pulmonary artery occlusion pressure from 14.6 +/- 1.6 mmHg to 17.4 +/- 1.7 mmHg (P < 0.001) and pulse pressure variation from 15.8 +/- 8.5% to 25.3 +/- 9.5% (P < 0.001). High tidal volume increased pulse pressure variation from 15.8 +/- 8.5% to 31.6 +/- 10.4% (P < 0.001), and an inspiratory to expiratory time ratio of 2: 1 significantly increased only central venous pressure. During hypovolaemia, high positive end-expiratory pressure influenced all studied variables, and high tidal volume strongly increased pulse pressure variation (40.5 +/- 12.4% pre vs. 84.2 +/- 19.1% post, P < 0.001). The inversion of the inspiratory to expiratory time ratio only slightly increased filling pressures during hypovolaemia, without affecting pulse pressure variation or right ventricle end-diastolic volume. CONCLUSION: We concluded that pulse pressure variation measurement is influenced by cyclic variations in intrathoracic pressure, such as those caused by augmentations in tidal volume. The increase in mean airway pressure caused by positive end-expiratory pressure affects cardiac filling pressures and also pulse pressure variation, although to a lesser extent. Inversion of the inspiratory to expiratory time ratio does not induce significant changes in static and functional haemodynamic parameters. PMID- 19122556 TI - Bolus dose remifentanil and sufentanil blunting cardiovascular intubation responses in children: a randomized, double-blind comparison. AB - BACKGROUND AND OBJECTIVE: The present randomized controlled study was designed to compare the efficacy of remifentanil 2 microg kg(-1) and sufentanil 0.2 microg kg(-1) by bolus injection on the cardiovascular response to intubation in healthy children. METHODS: One hundred and five children, ASA 1-2 and scheduled for elective plastic surgery under general anaesthesia, were randomly divided into one of two study groups to receive the following treatments in a double-blind manner: remifentanil 2 microg kg(-1) (group R) and sufentanil 0.2 microg kg(-1) (group S). Blood pressure and heart rate (HR) were recorded before anaesthesia induction (baseline values), immediately before intubation (postinduction values), at intubation and every minute for 5 min after intubation. The percentage changes in systolic blood pressure (SBP) and HR relative to baseline values were calculated. The incidences of SBP and HR percentage changes of more than 30% of baseline values during the observation were recorded. RESULTS: As compared with baseline values, blood pressure and HR at intubation and their maximum values during the observation increased significantly in group S, but decreased significantly in group R. Blood pressure and HR at intubation and their maximum values during the observation were significantly different between the groups. There were significant differences between the groups in the percentage changes of SBP and HR relative to baseline values and their maximum percentage changes during the observation. The incidences of SBP and HR percentage increases of more than 30% of baseline values were not significantly different between the groups, but the incidences of SBP and HR percentage decreases of more than 30% of baseline values were significantly higher in group R than in group S. CONCLUSION: In combination with propofol for anaesthesia induction in children, sufentanil 0.2 microg kg(-1) by bolus injection fails to depress the cardiovascular intubation response. Remifentanil 2 microg kg(-1) by bolus injection can completely abolish the cardiovascular intubation response, but causes more adverse cardiovascular depression. PMID- 19122558 TI - Fospropofol assay: issues and impact on pharmacokinetic and pharmacodynamic evaluation. PMID- 19122559 TI - Drugs used for spinal anaesthesia in patients undergoing ambulatory knee arthroscopy: a survey of Dutch anaesthesiologists. PMID- 19122560 TI - Kidney damage by hydroxyethyl starch of lower molecular weight and substitution. PMID- 19122561 TI - Sudden decrease in spectral entropy associated with severe carotid artery spasm. PMID- 19122562 TI - Intravenous dexmedetomidine infusion for labour analgesia in patient with preeclampsia. PMID- 19122563 TI - Two problems of empirical biomedical science: example of awareness monitoring with bispectral analysis. PMID- 19122564 TI - Anaesthesia and restless legs syndrome. PMID- 19122565 TI - Critical overfilling of a vaporizer. PMID- 19122566 TI - Long-acting beta-2 agonists in asthma. PMID- 19122568 TI - Silodosin (Rapaflo) for benign prostatic hyperplasia. AB - Another alpha1a blocker that claims to be more than just another alpha1 blocker. PMID- 19122567 TI - PPI interactions with clopidogrel. AB - Two drugs often taken together. Is there a problem? PMID- 19122569 TI - Citicoline: update on a promising and widely available agent for neuroprotection and neurorepair. AB - Choline precursors promote repair and growth of cell membranes and hold promise in a variety of neurologic diseases, including ischemic and hemorrhagic stroke. Citicoline, the most well-studied choline agent precursor, is widely prescribed throughout the world and recently became available in the United States as a dietary supplement. In experimental stroke models, citicoline conferred acute neuroprotection and enhanced neuroplasticity and neurorepair in the subacute period. Although individual human stroke trials have been inconclusive, meta analysis of 10 trials enrolling 2279 patients suggests patients receiving citicoline had substantially reduced frequencies of death and disability. Reinvestigation of citicoline with modern neuroimaging and clinical trial methods are underway and will provide more definitive information regarding the mechanistic and clinical effects of this promising neurotherapeutic agent. PMID- 19122570 TI - Antiepileptic drugs in aneurysmal subarachnoid hemorrhage. AB - Seizures may occur during or soon after rupture of an intracranial aneurysm. The use of antiepileptic drugs (AEDs) is a controversial issue. The overall conclusions from 2 recent studies in aneurysmal subarachnoid hemorrhage are that 1) many patients receive AEDs but should not; 2) long-term use is associated with worse outcome; and 3) short-term use is safer. Phenytoin may not be the first choice for seizure prophylaxis; newer AEDs such as levetiracetam might be more helpful in prevention and treatment of seizures. PMID- 19122571 TI - Prevention of sudden death in sleep. AB - Sudden death in sleep, particularly in the young, is a tragic occurrence that conjures thoughts of wrongdoing and foul play. Despite activation of powerful arousal mechanisms when a threat occurs, massive lesions, sudden insurmountable dysfunctions, or profound toxicities may lead to death. Sudden death in sleep is not always inexplicable or unavoidable, which opens a window of opportunity for its prevention. This review explores predisposing conditions and suspected risk factors of sudden death in sleep with a focus on prevention and management. PMID- 19122572 TI - Sleep-disordered breathing and stroke. AB - Sleep and stroke have an important and fascinating interaction. Patients with sleep-disordered breathing present with cardiovascular heart disease, cognitive decline, and increased risk of stroke. Stroke adversely affects sleep and factors such as prolonged immobilization, chronic pain, nocturnal hypoxia, and depression, which can also adversely impact sleep quality. Obstructive sleep apnea (OSA), one of the most common and serious sleep disturbances, manifests itself in almost 50% of all stroke patients. Sleep apnea patients who experience a stroke may be at a greater impairment in their rehabilitation potential and have increased risk of secondary stroke and mortality. Given these factors, the practicing neurologist should possess the skills to appropriately recognize, rapidly diagnose, and properly manage stroke patients with OSA. PMID- 19122573 TI - Stroke. PMID- 19122574 TI - A patient with subacute onset of ophthalmoplegia, papilledema, and proptosis. AB - A young man presented with headache, proptosis, and asymmetric ophthalmoplegia. He was found to have a mass in an unusual area of the brain. The differential diagnosis, diagnostic testing, pathology, treatment, and prognosis of this rare tumor are discussed in detail. PMID- 19122579 TI - Optimizing outcomes in coronary CT imaging. AB - Cardiac computed tomographic angiography (CCTA) is revolutionizing our approach to the identification and management of coronary artery atherosclerosis. CCTA has the unique ability to assess extracoronary cardiac findings within the standard field of view, including the anatomy of cardiac structures-the atria, ventricles, valves, pericardium, great vessels, and venous anatomy-and any related abnormalities. Appropriate clinical applications include evaluation of intermediate-risk patients with acute chest pain and those with suspected coronary anomalies, uninterpretable or equivocal stress test results, or suspected cardiac morphologic abnormalities. Optimization of this diagnostic examination requires close attention to details such as minimizing motion and attaining sufficient contrast opacification. Iso-osmolar contrast can help achieve maximum patient comfort with the smallest elevation of heart rate and variability, as well as minimize the risk of acute kidney injury. Newer scanning equipment and protocols have improved image quality in difficult cases, including obese patients and those with heavy coronary calcification or metal artifacts. Current imaging protocols have reduced exposure to ionizing radiation and continue to improve safety. PMID- 19122580 TI - Assessment of myocardial viability: review of the clinical significance. AB - The identification of myocardial viability in patients with coronary artery disease and left ventricular dysfunction (LVD) has important clinical and prognostic implications. Two terms commonly used to define clinical conditions of potentially reversible contractile dysfunction are stunned myocardium and hibernating myocardium. Stunned myocardium refers to transient depression of contractile function secondary to an acute ischemic insult. Hibernating myocardium is a form of contractile dysfunction of living myocytes in the setting of chronic ischemia or chronically reduced flow reserve. Numerous observational studies have shown improved clinical outcomes after revascularization of patients with LVD and evidence of myocardial viability, although patients with nonviable myocardium have not been shown to have the same benefits. The use of noninvasive techniques to determine myocardial viability provides important information to guide clinicians in deciding which patients with LVD are likely to receive benefit from a revascularization procedure. Positron emission tomography, single photon emission computed tomography, dobutamine echocardiography, and cardiac magnetic resonance imaging each have advantages and limitations. PMID- 19122581 TI - Magnetic resonance imaging in patients with implantable cardiac devices. AB - The use of magnetic resonance imaging (MRI) in patients with implantable cardiac devices, such as pacemakers, cardioverter defibrillators, and loop recorders, has been contraindicated based on concerns regarding the powerful magnetic field generated by MRI. Due to the widespread application and powerful diagnostic capability of MRI, there are instances in which denying a patient with an implantable cardiac device an MRI evaluation may influence the quality of health care received. There are data to suggest that MRI might be considered a relative contraindication instead of an absolute contraindication in device patients when precautions are taken by experienced physicians to lower the risk of adverse events. Despite the potential concerns, several hundred non-pacemaker-dependent patients and several pacemaker-dependent patients have undergone MRI without complications while being monitored under a number of different safety protocols. Various strategies have been used to minimize the risk of performing MRI procedures in device patients. Patient selection must be rigorous and made on a case-by-case basis. PMID- 19122582 TI - The importance of recognizing and treating low levels of high-density lipoprotein cholesterol: a new era in atherosclerosis management. AB - Low levels of high-density lipoprotein cholesterol (HDL-C) represent a major cardiovascular risk factor, with a stronger relationship to coronary heart disease than that seen with elevated levels of low-density lipoprotein cholesterol (LDL-C). HDL-C has important antiatherogenic effects, including reverse cholesterol transport, inhibition of LDL-C oxidation, and antiplatelet and anti-inflammatory actions. Patients with low HDL-C are also at an amplified risk of coronary heart disease due to the common coexistence of other risk factors, including excess adiposity, metabolic syndrome, type 2 diabetes mellitus, hypertriglyceridemia, and the atherogenic dyslipidemia characterized by small dense LDL-C. First-line therapy of low HDL-C generally consists of nonpharmacologic measures such as improved fitness and weight loss. Current pharmaceutical options include statins, fibrates, and nicotinic acid. A host of novel approaches involving HDL-C and reverse cholesterol transport hold the promise of fundamentally changing the natural history of atherosclerosis, the most common and important chronic disease in humans. PMID- 19122583 TI - The athlete's heart: prevention of sudden cardiac death. PMID- 19122586 TI - Symptomatic metastatic right atrial lymphoma in a patient with AIDS presenting with pulmonary embolization. AB - Tumors involving the heart are rare, and the majority of them are benign. Secondary lymphoma with localization to the heart is the third most common malignant heart tumor and is more common, by far, than primary cardiac lymphomas. In patients with human immunodeficiency virus, the risk of development of systemic lymphoma is 60 to 200 times higher than in the general population. Symptoms usually consist of chest pain and dyspnea. Patients can also present with obstructive symptoms, based on the location and size of the tumor, and signs such as elevated jugular venous pressure, peripheral edema, ascites, and hepatomegaly. Transthoracic echocardiography is the initial modality of choice for diagnosis of cardiac lymphomas because it is readily available and helps localize the tumor, but transesophageal echocardiography and magnetic resonance imaging remain the best tests for evaluation. Treatment consists primarily of chemotherapy, and anticoagulation can be used in certain cases where embolization of the tumor is likely. This case review describes a 37-year-old man with past medical history significant for herpes zoster and stage 1 syphilis who presented with complaints of weight loss, intermittent fevers, and vague chest pains of 1 month duration. PMID- 19122587 TI - Percutaneous coronary intervention. Cardiovascular outcomes and long-term clopidogrel use after PCI in patients with diabetes. PMID- 19122590 TI - Shaping embryos in Barcelona. AB - The Morphogenesis and Cell Behaviour meeting held this fall in Barcelona explored the role of forces, adhesion and oscillations in embryonic morphogenesis. It highlighted the impact of new microscopy methods and modelling approaches on the most recent advances of the field. PMID- 19122591 TI - Membrane repair redux: redox of MG53. PMID- 19122592 TI - Undressing a cellular corset: septins exposed. PMID- 19122593 TI - Ran out of the nucleus for apoptosis. PMID- 19122594 TI - sFRPs: a declaration of (Wnt) independence. PMID- 19122596 TI - Regulation of ERK activity duration by Sprouty contributes to dorsoventral patterning. AB - Distinct modes of ERK activation, sustained or transient, are essential for cell fate decision in cultured cells. Here we show that Xenopus laevis Sprouty2 (XSpry2) controls the duration of ERK activity and thereby contributes to the establishment of dorsoventral patterning during mesoderm formation. Furthermore, Xenopus Fos (XFos) can function as a molecular sensor of the ERK signalling duration in Xenopus embryos. This work provides the first evidence that regulating the duration of ERK activity contributes to cell fate decisions in the context of the whole organism. PMID- 19122601 TI - Experts still needed. PMID- 19122602 TI - Starry messages. PMID- 19122604 TI - A public service. PMID- 19122605 TI - Obama's picks underline climate focus. PMID- 19122606 TI - Universities struggle as value of endowments falls. PMID- 19122607 TI - Good grades, but who gets the cash? PMID- 19122608 TI - Single fund for African science aid. PMID- 19122616 TI - The new boss in town. PMID- 19122617 TI - Year of astronomy: New eyes, new skies. PMID- 19122618 TI - Honeybee and the Phoenix analysing instrument. PMID- 19122619 TI - Obama must match science rhetoric with action. PMID- 19122620 TI - Problems with anti-plagiarism database. PMID- 19122621 TI - Time to turn off the lights. PMID- 19122622 TI - Year of astronomy: Mankind's place in the Universe. PMID- 19122628 TI - Quantum physics: Squeeze until it hurts. PMID- 19122631 TI - Astrophysics: Star formation branches out. PMID- 19122629 TI - Cancer: Inflaming metastasis. PMID- 19122632 TI - Game theory: How to treat those of ill repute. PMID- 19122633 TI - Darwin 200: A natural selection. PMID- 19122634 TI - 18 years of science with the Hubble Space Telescope. AB - After several decades of planning, the Hubble Space Telescope (HST) was launched in 1990 as the first of NASA's Great Observatories. After a rocky start arising from an error in the fabrication of its main mirror, it went on to change forever many fields of astronomy, and to capture the public's imagination with its images. An ongoing programme of servicing missions has kept the telescope on the cutting edge of astronomical research. Here I review the advances made possible by the HST over the past 18 years. PMID- 19122636 TI - A role for self-gravity at multiple length scales in the process of star formation. AB - Self-gravity plays a decisive role in the final stages of star formation, where dense cores (size approximately 0.1 parsecs) inside molecular clouds collapse to form star-plus-disk systems. But self-gravity's role at earlier times (and on larger length scales, such as approximately 1 parsec) is unclear; some molecular cloud simulations that do not include self-gravity suggest that 'turbulent fragmentation' alone is sufficient to create a mass distribution of dense cores that resembles, and sets, the stellar initial mass function. Here we report a 'dendrogram' (hierarchical tree-diagram) analysis that reveals that self-gravity plays a significant role over the full range of possible scales traced by (13)CO observations in the L1448 molecular cloud, but not everywhere in the observed region. In particular, more than 90 per cent of the compact 'pre-stellar cores' traced by peaks of dust emission are projected on the sky within one of the dendrogram's self-gravitating 'leaves'. As these peaks mark the locations of already-forming stars, or of those probably about to form, a self-gravitating cocoon seems a critical condition for their existence. Turbulent fragmentation simulations without self-gravity-even of unmagnetized isothermal material-can yield mass and velocity power spectra very similar to what is observed in clouds like L1448. But a dendrogram of such a simulation shows that nearly all the gas in it (much more than in the observations) appears to be self-gravitating. A potentially significant role for gravity in 'non-self-gravitating' simulations suggests inconsistency in simulation assumptions and output, and that it is necessary to include self-gravity in any realistic simulation of the star formation process on subparsec scales. PMID- 19122635 TI - Cell-cycle restriction limits DNA damage and maintains self-renewal of leukaemia stem cells. AB - Rare cells with the properties of stem cells are integral to the development and perpetuation of leukaemias. A defining characteristic of stem cells is their capacity to self-renew, which is markedly extended in leukaemia stem cells. The underlying molecular mechanisms, however, are largely unknown. Here we demonstrate that expression of the cell-cycle inhibitor p21 is indispensable for maintaining self-renewal of leukaemia stem cells. Expression of leukaemia associated oncogenes in mouse haematopoietic stem cells (HSCs) induces DNA damage and activates a p21-dependent cellular response, which leads to reversible cell cycle arrest and DNA repair. Activated p21 is critical in preventing excess DNA damage accumulation and functional exhaustion of leukaemic stem cells. These data unravel the oncogenic potential of p21 and suggest that inhibition of DNA repair mechanisms might function as potent strategy for the eradication of the slowly proliferating leukaemia stem cells. PMID- 19122637 TI - Squeezing and over-squeezing of triphotons. AB - Quantum mechanics places a fundamental limit on the accuracy of measurements. In most circumstances, the measurement uncertainty is distributed equally between pairs of complementary properties; this leads to the 'standard quantum limit' for measurement resolution. Using a technique known as 'squeezing', it is possible to reduce the uncertainty of one desired property below the standard quantum limit at the expense of increasing that of the complementary one. Squeezing is already being used to enhance the sensitivity of gravity-wave detectors and may play a critical role in other high precision applications, such as atomic clocks and optical communications. Spin squeezing (the squeezing of angular momentum variables) is a powerful tool, particularly in the context of quantum light matter interfaces. Although impressive gains in squeezing have been made, optical spin-squeezed systems are still many orders of magnitude away from the maximum possible squeezing, known as the Heisenberg uncertainty limit. Here we demonstrate how an optical system can be squeezed essentially all the way to this fundamental bound. We construct spin-squeezed states by overlapping three indistinguishable photons in an optical fibre and manipulating their polarization (spin), resulting in the formation of a squeezed composite particle known as a 'triphoton'. The symmetry properties of polarization imply that the measured triphoton states can be most naturally represented by quasi-probability distributions on the surface of a sphere. In this work we show that the spherical topology of polarization imposes a limit on how much squeezing can occur, leading to the quasi-probability distributions wrapping around the sphere-a phenomenon we term 'over-squeezing'. Our observations of spin-squeezing in the few-photon regime could lead to new quantum resources for enhanced measurement, lithography and information processing that can be precisely engineered photon-by-photon. PMID- 19122638 TI - Optical manipulation of nanoparticles and biomolecules in sub-wavelength slot waveguides. AB - The ability to manipulate nanoscopic matter precisely is critical for the development of active nanosystems. Optical tweezers are excellent tools for transporting particles ranging in size from several micrometres to a few hundred nanometres. Manipulation of dielectric objects with much smaller diameters, however, requires stronger optical confinement and higher intensities than can be provided by these diffraction-limited systems. Here we present an approach to optofluidic transport that overcomes these limitations, using sub-wavelength liquid-core slot waveguides. The technique simultaneously makes use of near-field optical forces to confine matter inside the waveguide and scattering/adsorption forces to transport it. The ability of the slot waveguide to condense the accessible electromagnetic energy to scales as small as 60 nm allows us also to overcome the fundamental diffraction problem. We apply the approach here to the trapping and transport of 75-nm dielectric nanoparticles and lambda-DNA molecules. Because trapping occurs along a line, rather than at a point as with traditional point traps, the method provides the ability to handle extended biomolecules directly. We also carry out a detailed numerical analysis that relates the near-field optical forces to release kinetics. We believe that the architecture demonstrated here will help to bridge the gap between optical manipulation and nanofluidics. PMID- 19122639 TI - Seismic evidence for overpressured subducted oceanic crust and megathrust fault sealing. AB - Water and hydrous minerals play a key part in geodynamic processes at subduction zones by weakening the plate boundary, aiding slip and permitting subduction-and indeed plate tectonics-to occur. The seismological signature of water within the forearc mantle wedge is evident in anomalies with low seismic shear velocity marking serpentinization. However, seismological observations bearing on the presence of water within the subducting plate itself are less well documented. Here we use converted teleseismic waves to obtain observations of anomalously high Poisson's ratios within the subducted oceanic crust from the Cascadia continental margin to its intersection with forearc mantle. On the basis of pressure, temperature and compositional considerations, the elevated Poisson's ratios indicate that water is pervasively present in fluid form at pore pressures near lithostatic values. Combined with observations of a strong negative velocity contrast at the top of the oceanic crust, our results imply that the megathrust is a low-permeability boundary. The transition from a low- to high-permeability plate interface downdip into the mantle wedge is explained by hydrofracturing of the seal by volume changes across the interface caused by the onset of crustal eclogitization and mantle serpentinization. These results may have important implications for our understanding of seismogenesis, subduction zone structure and the mechanism of episodic tremor and slip. PMID- 19122640 TI - Indirect reciprocity provides only a narrow margin of efficiency for costly punishment. AB - Indirect reciprocity is a key mechanism for the evolution of human cooperation. Our behaviour towards other people depends not only on what they have done to us but also on what they have done to others. Indirect reciprocity works through reputation. The standard model of indirect reciprocity offers a binary choice: people can either cooperate or defect. Cooperation implies a cost for the donor and a benefit for the recipient. Defection has no cost and yields no benefit. Currently there is considerable interest in studying the effect of costly (or altruistic) punishment on human behaviour. Punishment implies a cost for the punished person. Costly punishment means that the punisher also pays a cost. It has been suggested that costly punishment between individuals can promote cooperation. Here we study the role of costly punishment in an explicit model of indirect reciprocity. We analyse all social norms, which depend on the action of the donor and the reputation of the recipient. We allow errors in assigning reputation and study gossip as a mechanism for establishing coherence. We characterize all strategies that allow the evolutionary stability of cooperation. Some of those strategies use costly punishment; others do not. We find that punishment strategies typically reduce the average payoff of the population. Consequently, there is only a small parameter region where costly punishment leads to an efficient equilibrium. In most cases the population does better by not using costly punishment. The efficient strategy for indirect reciprocity is to withhold help for defectors rather than punishing them. PMID- 19122641 TI - Carcinoma-produced factors activate myeloid cells through TLR2 to stimulate metastasis. AB - Metastatic progression depends on genetic alterations intrinsic to cancer cells as well as the inflammatory microenvironment of advanced tumours. To understand how cancer cells affect the inflammatory microenvironment, we conducted a biochemical screen for macrophage-activating factors secreted by metastatic carcinomas. Here we show that, among the cell lines screened, Lewis lung carcinoma (LLC) were the most potent macrophage activators leading to production of interleukin-6 (IL-6) and tumour-necrosis factor-alpha (TNF-alpha) through activation of the Toll-like receptor (TLR) family members TLR2 and TLR6. Both TNF alpha and TLR2 were found to be required for LLC metastasis. Biochemical purification of LLC-conditioned medium (LCM) led to identification of the extracellular matrix proteoglycan versican, which is upregulated in many human tumours including lung cancer, as a macrophage activator that acts through TLR2 and its co-receptors TLR6 and CD14. By activating TLR2:TLR6 complexes and inducing TNF-alpha secretion by myeloid cells, versican strongly enhances LLC metastatic growth. These results explain how advanced cancer cells usurp components of the host innate immune system, including bone-marrow-derived myeloid progenitors, to generate an inflammatory microenvironment hospitable for metastatic growth. PMID- 19122642 TI - Chaperonin complex with a newly folded protein encapsulated in the folding chamber. AB - A subset of essential cellular proteins requires the assistance of chaperonins (in Escherichia coli, GroEL and GroES), double-ring complexes in which the two rings act alternately to bind, encapsulate and fold a wide range of nascent or stress-denatured proteins. This process starts by the trapping of a substrate protein on hydrophobic surfaces in the central cavity of a GroEL ring. Then, binding of ATP and co-chaperonin GroES to that ring ejects the non-native protein from its binding sites, through forced unfolding or other major conformational changes, and encloses it in a hydrophilic chamber for folding. ATP hydrolysis and subsequent ATP binding to the opposite ring trigger dissociation of the chamber and release of the substrate protein. The bacteriophage T4 requires its own version of GroES, gp31, which forms a taller folding chamber, to fold the major viral capsid protein gp23 (refs 16-20). Polypeptides are known to fold inside the chaperonin complex, but the conformation of an encapsulated protein has not previously been visualized. Here we present structures of gp23-chaperonin complexes, showing both the initial captured state and the final, close-to-native state with gp23 encapsulated in the folding chamber. Although the chamber is expanded, it is still barely large enough to contain the elongated gp23 monomer, explaining why the GroEL-GroES complex is not able to fold gp23 and showing how the chaperonin structure distorts to enclose a large, physiological substrate protein. PMID- 19122644 TI - Chemokine-mediated migration of skin-derived stem cells: predominant role for CCL5/RANTES. AB - The ability of stem cells to self-renew as well as their multilineage differentiation potential makes them ideal candidates for skin regeneration strategies. Mesenchymal stem cells residing in human adult dermis, in contrast to adipose tissue, have not yet been described. The objective of this study was to determine the stemness and chemokine-mediated homing potential of dermal stromal cells (DSC) and to compare this with adipose stem cells (ASC). DSC have a less stellate form than ASC, confirming that DSC and ASC are two different types of mesenchymal cell populations. However, DSC display a mesenchymal stem cell phenotype (CD31(-), CD34(+), CD45(-), CD54(+), CD90(+), CD105(+), and CD166(+) similar to ASC and are also multipotent in their ability to differentiate into adipocytes, chondrocytes, and osteoblasts. Both ASC and DSC display a similar set of chemokine receptors (CCR3, CCR4, CCR6, CCR10, CXCR1, and CXCR2). Several ligands for these receptors, with CCL5/RANTES being the most potent, can induce migration of ASC and DSC in an in vitro wound-healing assay. Taken together, these results show that a population of mesenchymal stem cells resides in the dermis of human adult skin and these dermal-derived stem cells have a phenotypic and chemokine-mediated homing potential similar to adipose stem cells, which to our knowledge is previously unreported. PMID- 19122645 TI - Functional expression of the intracellular pattern recognition receptor NOD1 in human keratinocytes. PMID- 19122646 TI - Development of an ichthyosiform phenotype in Alox12b-deficient mouse skin transplants. AB - 12R-lipoxygenase (12R-LOX) represents a key enzyme of a recently identified eicosanoid pathway in the skin that plays an essential role in the establishment and/or maintenance of the epidermal barrier function. Genetic studies show that loss-of-function mutations in ALOX12B, encoding 12R-LOX, and in ALOXE3, encoding another closely related LOX involved in this pathway, are the second most common cause for autosomal recessive congenital ichthyosis (ARCI). To investigate the pathomechanism of ARCI and the function of 12R-LOX, we recently generated a 12R LOX knockout model. 12R-LOX-deficient mice die rapidly after birth from severe barrier dysfunction without exhibiting an obvious cutaneous phenotype. Thus, we analyzed the adult phenotype of 12R-LOX(-/-) skin transplanted onto nude mice. 12R-LOX(-/-) skin develops an ichthyosiform appearance with thickening of the epidermis, hyperproliferation, hypergranulosis, focal parakeratosis, and severe hyperkeratosis. The adult mutant mouse skin phenotype closely reproduces the ichthyosis phenotype seen in patients with ALOX12B mutations. Western blot analysis revealed restoration of profilaggrin processing that used to be disturbed in neonatal mutant skin and overexpression of filaggrin, involucrin, and repetin. The results indicate that 12R-LOX knockout mice may represent a useful animal model for a detailed analysis of mechanisms involved in ARCI forms that are associated with impaired LOX metabolism. PMID- 19122647 TI - Loss of class III beta-tubulin induced by histone deacetylation is associated with chemosensitivity to paclitaxel in malignant melanoma cells. AB - Overexpression of class III beta-tubulin (TUBB3) is an important mechanism of taxane resistance. Using 7 melanoma cell lines, 2 normal neonatal human epidermal melanocyte (NHEM) cultures, and 49 primary melanomas, we investigated TUBB3 expression, its relationship to chemosensitivity to taxane derivatives, and the epigenetic mechanism controlling TUBB3 gene expression. Normal melanocytes in vitro and in vivo strongly expressed TUBB3 protein. NHEMs exhibited marked chemoresistance to paclitaxel-induced apoptosis. A subset (10 of 49, 20%) of primary malignant melanomas was TUBB3 negative. The incidence of TUBB3-negative melanomas increased with stage of progression. TUBB3 protein expression varied among cell lines; one (HMV-I) of the seven cell lines exhibited an extremely low endogenous level. TUBB3 protein expression correlated well with chemosensitivity to paclitaxel-induced apoptosis (P<0.05). Treatment with a histone deacetylase (HDAC) inhibitor restored TUBB3 expression in HMV-I. Chromatin immunoprecipitation assays revealed that histones H3 and H4 were hypoacetylated at the TUBB3 gene in HMV-I as compared with a TUBB3-overexpressing cell type (HMV II). Treatment with the HDAC inhibitor induced gain of histone acetylation only in HMV-I. These results suggest that loss of TUBB3 protein may be induced by histone deacetylation in a subset of malignant melanomas, and may be associated with chemosensitivity to taxane. PMID- 19122648 TI - Azathioprine-induced microsatellite instability is not observed in skin carcinomas of organ transplant recipients. PMID- 19122649 TI - Elevated dietary magnesium prevents connective tissue mineralization in a mouse model of pseudoxanthoma elasticum (Abcc6(-/-)). AB - Pseudoxanthoma elasticum (PXE) is an autosomal recessive multisystem disorder characterized by ectopic connective tissue mineralization, with clinical manifestations primarily in the skin, eyes, and cardiovascular system. There is considerable, both intra- and interfamilial, variability in the spectrum of phenotypic presentation. Previous studies have suggested that mineral content of the diet may modify the severity of the clinical phenotype in PXE. In this study, we utilized a targeted mutant mouse (Abcc6(-/-)) as a model system for PXE. We examined the effects of changes in dietary phosphate and magnesium on the mineralization process using calcification of the connective tissue capsule surrounding the vibrissae as an early phenotypic biomarker. Mice placed on custom designed diets either high or low in phosphate did not show changes in mineralization, which was similar to that noted in Abcc6(-/-) mice on control diet. However, mice placed on diet enriched in magnesium (fivefold) showed no evidence of connective tissue mineralization in this mouse model of PXE. The inhibitory capacity of magnesium was confirmed in a cell-based mineralization assay system in vitro. Collectively, our observations suggest that assessment of dietary magnesium in patients with PXE may be warranted. PMID- 19122650 TI - Increased hyaluronan production and decreased E-cadherin expression by cytokine stimulated keratinocytes lead to spongiosis formation. AB - The pathogenesis of spongiosis, which is a well-known hallmark of acute eczema, is not fully understood. We sought to clarify the mechanism for the influx of tissue fluid into the epidermis and the loss of cohesion between keratinocytes in acute eczema that result in spongiosis. We first demonstrated increased intercellular accumulation of hyaluronan (HA) in the spongiotic epidermis by immunochemical staining using hyaluronic-acid-binding protein (HABP) and augmented hyaluronan synthase 3 (HAS3) mRNA expression by spongiotic keratinocytes using in situ hybridization. We also showed that the epidermis where the intercellular space was strongly stained with HABP showed weaker expression of membrane E-cadherin. Next, we demonstrated--by a sandwich assay using HABP, real-time PCR, and flow cytometry--that, among various cytokines, only IL-4, IL-13, and IFN-gamma increased HA production, enhanced HAS3 mRNA expression, and decreased membrane E-cadherin expression by normal human epidermal keratinocytes in both low- and high-Ca media. Finally, we demonstrated IL-4, IL-13, their combination, and IFN-gamma could induce intercellular space widening of the epidermis with increased HA accumulation and decreased E-cadherin expression in the organotypic culture. These results suggest that the augmented production of HA and the decreased E-cadherin expression by keratinocytes stimulated with IL-4/IL-13 or IFN-gamma cause spongiosis in acute eczema. PMID- 19122651 TI - PeakSeq enables systematic scoring of ChIP-seq experiments relative to controls. AB - Chromatin immunoprecipitation (ChIP) followed by tag sequencing (ChIP-seq) using high-throughput next-generation instrumentation is fast, replacing chromatin immunoprecipitation followed by genome tiling array analysis (ChIP-chip) as the preferred approach for mapping of sites of transcription-factor binding and chromatin modification. Using two deeply sequenced data sets for human RNA polymerase II and STAT1, each with matching input-DNA controls, we describe a general scoring approach to address unique challenges in ChIP-seq data analysis. Our approach is based on the observation that sites of potential binding are strongly correlated with signal peaks in the control, likely revealing features of open chromatin. We develop a two-pass strategy called PeakSeq to compensate for this. A two-pass strategy compensates for signal caused by open chromatin, as revealed by inclusion of the controls. The first pass identifies putative binding sites and compensates for genomic variation in the 'mappability' of sequences. The second pass filters out sites not significantly enriched compared to the normalized control, computing precise enrichments and significances. Our scoring procedure enables us to optimize experimental design by estimating the depth of sequencing required for a desired level of coverage and demonstrating that more than two replicates provides only a marginal gain in information. PMID- 19122652 TI - Characterization of human embryonic stem cells with features of neoplastic progression. AB - Cultured human embryonic stem (hES) cells can acquire genetic and epigenetic changes that make them vulnerable to transformation. As hES cells with cancer cell characteristics share properties with normal hES cells, such as self renewal, teratoma formation and the expression of pluripotency markers, they may be misconstrued as superior hES cells with enhanced 'stemness'. We characterize two variant hES cell lines (v-hESC-1 and v-hESC-2) that express pluripotency markers at high levels and do not harbor chromosomal abnormalities by standard cytogenetic measures. We show that the two lines possess some features of neoplastic progression, including a high proliferative capacity, growth-factor independence, a 9- to 20-fold increase in frequency of tumor-initiating cells, niche independence and aberrant lineage specification, although they are not malignant. Array comparative genomic hybridization reveals an amplification at 20q11.1-11.2 in v-hESC-1 and a deletion at 5q34a-5q34b;5q3 and a mosaic gain of chromosome 12 in v-hESC-2. These results emphasize the need for functional characterization to distinguish partially transformed and normal hES cells. PMID- 19122653 TI - Dectin-1 directs T helper cell differentiation by controlling noncanonical NF kappaB activation through Raf-1 and Syk. AB - The C-type lectin dectin-1 activates the transcription factor NF-kappaB through a Syk kinase-dependent signaling pathway to induce antifungal immunity. Here we show that dectin-1 expressed on human dendritic cells activates not only the Syk dependent canonical NF-kappaB subunits p65 and c-Rel, but also the noncanonical NF-kappaB subunit RelB. Dectin-1, when stimulated by the beta-glucan curdlan or by Candida albicans, induced a second signaling pathway mediated by the serine threonine kinase Raf-1, which integrated with the Syk pathway at the point of NF kappaB activation. Raf-1 antagonized Syk-induced RelB activation by promoting sequestration of RelB into inactive p65-RelB dimers, thereby altering T helper cell differentiation. Thus, dectin-1 activates two independent signaling pathways, one through Syk and one through Raf-1, to induce immune responses. PMID- 19122654 TI - The kinase PDK1 integrates T cell antigen receptor and CD28 coreceptor signaling to induce NF-kappaB and activate T cells. AB - In addition to ligation of the T cell antigen receptor (TCR), activation of the CD28 coreceptor by the costimulatory molecule B7 is required for induction of the transcription factor NF-kappaB and robust T cell activation, although the contribution of CD28 to this process remains incompletely understood. We show here that phosphoinositide-dependent kinase 1 (PDK1) is essential for integrating the TCR and CD28 signals. After we deleted PDK1 from T cells, TCR-CD28 signals were unable to induce activation of NF-kappaB or phosphorylation of protein kinase C-theta, although T cell survival and pathways dependent on the kinases p38 and Jnk or the transcription factor NFAT were unaffected. CD28 facilitated NF kappaB activation by regulating recruitment and phosphorylation of PDK1, which are necessary for efficient binding of PDK1 to protein kinase C-theta and the adaptor CARMA1 and thus for NF-kappaB induction. PMID- 19122655 TI - Hypoxia-inducible factor-dependent induction of netrin-1 dampens inflammation caused by hypoxia. AB - The neuronal guidance molecule netrin-1 is linked to the coordination of inflammatory responses. Given that mucosal surfaces are particularly prone to hypoxia-elicited inflammation, we sought to determine the function of netrin-1 in hypoxia-induced inflammation. We detected hypoxia-inducible factor 1alpha (HIF 1alpha)-dependent induction of expression of the gene encoding netrin-1 (Ntn1) in hypoxic epithelia. Neutrophil transepithelial migration studies showed that by engaging A2B adenosine receptor (A2BAR) on neutrophils, netrin-1 attenuated neutrophil transmigration. Exogenous netrin-1 suppressed hypoxia-elicited inflammation in wild-type but not in A2BAR-deficient mice, and inflammatory hypoxia was enhanced in Ntn1(+/-) mice relative to that in Ntn1(+/+) mice. Our studies demonstrate that HIF-1alpha-dependent induction of netrin-1 attenuates hypoxia-elicited inflammation at mucosal surfaces. PMID- 19122656 TI - Decreased levels of microRNA miR-122 in individuals with hepatitis C responding poorly to interferon therapy. AB - Several microRNAs (miRNAs), including liver-specific miR-122, have been implicated in the control of hepatitis C virus (HCV) RNA replication and its response to interferon (IFN) in human hepatoma cells. Our analysis of liver biopsies from subjects with chronic hepatitis C (CHC) undergoing IFN therapy revealed no correlation of miR-122 expression with viral load and markedly decreased pretreatment miR-122 levels in subjects who had no virological response during later IFN therapy; other investigated miRNAs showed only limited changes. These data have implications for the prospect of targeting miRNAs for CHC therapy. PMID- 19122657 TI - Disrupting functional interactions between platelet chemokines inhibits atherosclerosis in hyperlipidemic mice. AB - Atherosclerosis is characterized by chronic inflammation of the arterial wall due to chemokine-driven mononuclear cell recruitment. Activated platelets can synergize with chemokines to exacerbate atherogenesis; for example, by deposition of the chemokines platelet factor-4 (PF4, also known as CXCL4) and RANTES (CCL5), triggering monocyte arrest on inflamed endothelium. Homo-oligomerization is required for the recruitment functions of CCL5, and chemokine heteromerization has more recently emerged as an additional regulatory mechanism, as evidenced by a mutual modulation of CXCL8 and CXCL4 activities and by enhanced monocyte arrest resulting from CCL5-CXCL4 interactions. The CCL5 antagonist Met-RANTES reduces diet-induced atherosclerosis; however, CCL5 antagonism may not be therapeutically feasible, as suggested by studies using Ccl5-deficient mice which imply that direct CCL5 blockade would severely compromise systemic immune responses, delay macrophage-mediated viral clearance and impair normal T cell functions. Here we determined structural features of CCL5-CXCL4 heteromers and designed stable peptide inhibitors that specifically disrupt proinflammatory CCL5-CXCL4 interactions, thereby attenuating monocyte recruitment and reducing atherosclerosis without the aforementioned side effects. These results establish the in vivo relevance of chemokine heteromers and show the potential of targeting heteromer formation to achieve therapeutic effects. PMID- 19122658 TI - A stroma-related gene signature predicts resistance to neoadjuvant chemotherapy in breast cancer. AB - To better understand the relationship between tumor-host interactions and the efficacy of chemotherapy, we have developed an analytical approach to quantify several biological processes observed in gene expression data sets. We tested the approach on tumor biopsies from individuals with estrogen receptor-negative breast cancer treated with chemotherapy. We report that increased stromal gene expression predicts resistance to preoperative chemotherapy with 5-fluorouracil, epirubicin and cyclophosphamide (FEC) in subjects in the EORTC 10994/BIG 00-01 trial. The predictive value of the stromal signature was successfully validated in two independent cohorts of subjects who received chemotherapy but not in an untreated control group, indicating that the signature is predictive rather than prognostic. The genes in the signature are expressed in reactive stroma, according to reanalysis of data from microdissected breast tumor samples. These findings identify a previously undescribed resistance mechanism to FEC treatment and suggest that antistromal agents may offer new ways to overcome resistance to chemotherapy. PMID- 19122659 TI - Development of a novel mouse glioma model using lentiviral vectors. AB - We report the development of a new method to induce glioblastoma multiforme in adult immunocompetent mice by injecting Cre-loxP-controlled lentiviral vectors expressing oncogenes. Cell type- or region-specific expression of activated forms of the oncoproteins Harvey-Ras and AKT in fewer than 60 glial fibrillary acidic protein-positive cells in the hippocampus, subventricular zone or cortex of mice heterozygous for the gene encoding the tumor suppressor Tp53 were tested. Mice developed glioblastoma multiforme when transduced either in the subventricular zone or the hippocampus. However, tumors were rarely detected when the mice were transduced in the cortex. Transplantation of brain tumor cells into naive recipient mouse brain resulted in the formation of glioblastoma multiforme-like tumors, which contained CD133(+) cells, formed tumorspheres and could differentiate into neurons and astrocytes. We suggest that the use of Cre-loxP controlled lentiviral vectors is a novel way to generate a mouse glioblastoma multiforme model in a region- and cell type-specific manner in adult mice. PMID- 19122660 TI - Ligands bind to Sortilin in the tunnel of a ten-bladed beta-propeller domain. AB - The structure of the Sortilin ectodomain in complex with neurotensin has been determined at 2-A resolution, revealing that the C-terminal part of neurotensin binds in the tunnel of a ten-bladed beta-propeller domain. Binding competition studies suggest that additional binding sites, for example, for the prodomain of nerve growth factor-beta, are present in the tunnel and that competition for binding relates to the restricted space inside the propeller. PMID- 19122661 TI - The mechanism of pentabromopseudilin inhibition of myosin motor activity. AB - We have identified pentabromopseudilin (PBP) as a potent inhibitor of myosin dependent processes such as isometric tension development and unloaded shortening velocity. PBP-induced reductions in the rate constants for ATP binding, ATP hydrolysis and ADP dissociation extend the time required per myosin ATPase cycle in the absence and presence of actin. Additionally, coupling between the actin and nucleotide binding sites is reduced in the presence of the inhibitor. The selectivity of PBP differs from that observed with other myosin inhibitors. To elucidate the binding mode of PBP, we crystallized the Dictyostelium myosin-2 motor domain in the presence of Mg(2+)-ADP-meta-vanadate and PBP. The electron density for PBP is unambiguous and shows PBP to bind at a previously unknown allosteric site near the tip of the 50-kDa domain, at a distance of 16 A from the nucleotide binding site and 7.5 A away from the blebbistatin binding pocket. PMID- 19122662 TI - A transcriptome atlas of rice cell types uncovers cellular, functional and developmental hierarchies. AB - The functions of the plant body rely on interactions among distinct and nonequivalent cell types. The comparison of transcriptomes from different cell types should expose the transcriptional networks that underlie cellular attributes and contributions. Using laser microdissection and microarray profiling, we have produced a cell type transcriptome atlas that includes 40 cell types from rice (Oryza sativa) shoot, root and germinating seed at several developmental stages, providing patterns of cell specificity for individual genes and gene classes. Cell type comparisons uncovered previously unrecognized properties, including cell-specific promoter motifs and coexpressed cognate binding factor candidates, interaction partner candidates and hormone response centers. We inferred developmental regulatory hierarchies of gene expression in specific cell types by comparison of several stages within root, shoot and embryo. PMID- 19122663 TI - Loss-of-function mutations of an inhibitory upstream ORF in the human hairless transcript cause Marie Unna hereditary hypotrichosis. AB - Marie Unna hereditary hypotrichosis (MUHH) is an autosomal dominant form of genetic hair loss. In a large Chinese family carrying MUHH, we identified a pathogenic initiation codon mutation in U2HR, an inhibitory upstream ORF in the 5' UTR of the gene encoding the human hairless homolog (HR). U2HR is predicted to encode a 34-amino acid peptide that is highly conserved among mammals. In 18 more families from different ancestral groups, we identified a range of defects in U2HR, including loss of initiation, delayed termination codon and nonsense and missense mutations. Functional analysis showed that these classes of mutations all resulted in increased translation of the main HR physiological ORF. Our results establish the link between MUHH and U2HR, show that fine-tuning of HR protein levels is important in control of hair growth, and identify a potential mechanism for preventing hair loss or promoting hair removal. PMID- 19122664 TI - Ulcerative colitis-risk loci on chromosomes 1p36 and 12q15 found by genome-wide association study. AB - Ulcerative colitis is a chronic inflammatory disease of the colon that presents as diarrhea and gastrointestinal bleeding. We performed a genome-wide association study using DNA samples from 1,052 individuals with ulcerative colitis and preexisting data from 2,571 controls, all of European ancestry. In an analysis that controlled for gender and population structure, ulcerative colitis loci attaining genome-wide significance and subsequent replication in two independent populations were identified on chromosomes 1p36 (rs6426833, combined P = 5.1 x 10(-13), combined odds ratio OR = 0.73) and 12q15 (rs1558744, combined P = 2.5 x 10(-12), combined OR = 1.35). In addition, combined genome-wide significant evidence for association was found in a region spanning BTNL2 to HLA-DQB1 on chromosome 6p21 (rs2395185, combined P = 1.0 x 10(-16), combined OR = 0.66) and at the IL23R locus on chromosome 1p31 (rs11209026, combined P = 1.3 x 10(-8), combined OR = 0.56; rs10889677, combined P = 1.3 x 10(-8), combined OR = 1.29). PMID- 19122665 TI - Wnt antagonism of Shh facilitates midbrain floor plate neurogenesis. AB - The floor plate, an essential ventral midline organizing center that produces the morphogen Shh, has distinct properties along the neuraxis. The neurogenic potential of the floor plate and its underlying mechanisms remain unknown. Using Shh as a driver for lineage analysis, we found that the mouse midbrain, but not the hindbrain, floor plate is neurogenic, giving rise to dopamine (DA) neurons. Distinct spatiotemporal Shh and Wnt expression may distinguish the neurogenetic potential of these structures. We discovered an inhibitory role for Shh: removal of Shh resulted in neurogenesis from the hindbrain midline and, conversely, high doses of Shh inhibited proliferation and DA neuron production in midbrain cultures. We found that Wnt/beta-catenin signaling is necessary and sufficient for antagonizing Shh, DA progenitor marker induction and promotion of dopaminergic neurogenesis. These findings demonstrate how the dynamic interplay of canonical Wnt/beta-catenin signaling and Shh may orchestrate floor plate neurogenesis or a lack thereof. PMID- 19122666 TI - Collagen VI protects neurons against Abeta toxicity. AB - Amyloid-beta (Abeta) peptides, widely presumed to cause Alzheimer's disease, increased mouse neuronal expression of collagen VI through a mechanism involving transforming growth factor signaling. Reduction of collagen VI augmented Abeta neurotoxicity, whereas treatment of neurons with soluble collagen VI blocked the association of Abeta oligomers with neurons, enhanced Abeta aggregation and prevented neurotoxicity. These results identify collagen VI as an important component of the neuronal injury response and demonstrate its neuroprotective potential. PMID- 19122667 TI - Genetically timed, activity-sensor and rainbow transsynaptic viral tools. AB - We developed retrograde, transsynaptic pseudorabies viruses (PRVs) with genetically encoded activity sensors that optically report the activity of connected neurons among spatially intermingled neurons in the brain. Next we engineered PRVs to express two differentially colored fluorescent proteins in a time-shifted manner to define a time period early after infection to investigate neural activity. Finally we used multiple-colored PRVs to differentiate and dissect the complex architecture of brain regions. PMID- 19122668 TI - Microfluidic control of cell pairing and fusion. AB - Cell fusion has been used for many different purposes, including generation of hybridomas and reprogramming of somatic cells. The fusion step is the key event in initiation of these procedures. Standard fusion techniques, however, provide poor and random cell contact, leading to low yields. We present here a microfluidic device to trap and properly pair thousands of cells. Using this device, we paired different cell types, including fibroblasts, mouse embryonic stem cells and myeloma cells, achieving pairing efficiencies up to 70%. The device is compatible with both chemical and electrical fusion protocols. We observed that electrical fusion was more efficient than chemical fusion, with membrane reorganization efficiencies of up to 89%. We achieved greater than 50% properly paired and fused cells over the entire device, fivefold greater than with a commercial electrofusion chamber and observed reprogramming in hybrids between mouse embryonic stem cells and mouse embryonic fibroblasts. PMID- 19122669 TI - A genetically encoded fluorescent reporter of ATP:ADP ratio. AB - We constructed a fluorescent sensor of adenylate nucleotides by combining a circularly permuted variant of GFP with a bacterial regulatory protein, GlnK1, from Methanococcus jannaschii. The sensor's affinity for Mg-ATP was <100 nM, as seen for other members of the bacterial PII regulator family, a surprisingly high affinity given that normal intracellular ATP concentration is in the millimolar range. ADP bound the same site of the sensor as Mg-ATP, competing with it, but produced a smaller change in fluorescence. At physiological ATP and ADP concentrations, the binding site is saturated, but competition between the two substrates causes the sensor to behave as a nearly ideal reporter of the ATP:ADP concentration ratio. This principle for sensing the ratio of two analytes by competition at a high-affinity site probably underlies the normal functioning of PII regulatory proteins. The engineered sensor, Perceval, can be used to monitor the ATP:ADP ratio during live-cell imaging. PMID- 19122670 TI - Involvement of the ipsilateral and contralateral cerebellum in the acquisition of unilateral classical eyeblink conditioning in guinea pigs. AB - AIM: The aim of this study was to evaluate the relative contributions of the ipsilateral and contralateral cerebellum to the acquisition of unilateral classical eyeblink conditioning (EBCC). METHODS: The unilateral EBCC was achieved using a binaural tone conditioned stimulus (CS) paired with a left airpuff unconditioned stimulus (US). A high-resolution potentiometer was used to monitor eyeblink responses. Guinea pigs received one CS-US session followed by three CS US sessions (sessions 2 to 4), during which microinjections of muscimol, a GABA(A) receptor agonist, were performed to reversibly inactivate the cerebellum unilaterally prior to training. To test whether any learning had occurred during these inactivation sessions, training was continued for six more CS-US sessions (sessions 5 to 10) without any inactivation. RESULTS: Animals with inactivation of the left cerebellum had no signs of left conditioned response (CR) during sessions 2 to 4, and their CR acquisition during sessions 5 to 10 was not distinguishable from that of control animals during sessions 2 to 7. In contrast, animals with inactivation of the right cerebellum acquired left CRs during sessions 2 to 4, although their CR acquisition was significantly retarded during session 2. In addition, microinjections of muscimol into the right cerebellum did not affect left neuro-behavioral activity. Finally, microinjections of muscimol into either the left or the right cerebellum did not affect the performance of tone-airpuff evoked unconditioned response (UR). CONCLUSION: In contrast to the essential role of the ipsilateral cerebellum, the contralateral cerebellum is potentially involved in the acquisition of unilateral EBCC during the early stage of training. PMID- 19122671 TI - Effect of geniposide, a hypoglycemic glucoside, on hepatic regulating enzymes in diabetic mice induced by a high-fat diet and streptozotocin. AB - AIM: Hepatic glycogen phosphorylase (GP) and glucose-6-phosphatase (G6Pase) play an important role in the control of blood glucose homeostasis and are proposed to be potential targets for anti-diabetic drugs. Geniposide is an iridoid glucoside extracted from Gardenia jasminoides Ellis fruits and has been reported to have a hypoglycemic effect. However, little is known about the biochemical mechanisms by which geniposide regulates hepatic glucose-metabolizing enzymes. The present study investigates whether the hypoglycemic effect of geniposide is mediated by GP or G6Pase. METHODS: Type 2 diabetic mice, induced by a high-fat diet and streptozotocin injection, were treated with or without geniposide for 2 weeks. Blood glucose levels were monitored by a glucometer. Insulin concentrations were analyzed by the ELISA method. Total cholesterol (TC) and triglyceride (TG) levels were measured using Labassay kits. Activities of hepatic GP and G6Pase were measured by glucose-6-phosphate dehydrogenase-coupled reaction. Real-time RT-PCR and Western blotting were used to determine the mRNA and protein levels of both enzymes. RESULTS: Geniposide (200 and 400 mg/kg) significantly decreased the blood glucose, insulin and TG levels in diabetic mice in a dose-dependent manner. This compound also decreased the expression of GP and G6Pase at mRNA and immunoreactive protein levels, as well as enzyme activity. CONCLUSION: Geniposide is an effective hypoglycemic agent in diabetic mice. The hypoglycemic effect of this compound may be mediated, at least in part, by inhibiting the GP and G6Pase activities. PMID- 19122672 TI - Heme oxygenase-1 mediates the anti-inflammatory effect of isoflurane preconditioning in LPS-stimulated macrophages. AB - AIM: The aim of this study was to investigate the anti-inflammatory action of isoflurane preconditioning in a model of lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophages and examine the role of heme oxygenase (HO) 1 in this process. METHODS: Murine 264.7 macrophages were pretreated with or without 1%-3% isoflurane for 1 h. Thirty minutes later, the cells were incubated with or without LPS for 24 h. Cell viability was assessed using a 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell injury was assessed by measuring the release of lactate dehydrogenase (LDH). HO-1 and inducible nitric oxide synthase (iNOS) protein expression was analyzed by Western blotting. Tumor necrosis factor (TNF)-alpha levels, nitrite production and HO activity were also determined. RESULTS: Pretreatment with the nontoxic and clinically approved anesthetic isoflurane potently attenuated the cell injury and the decrease in cell viability that was induced by LPS. Treatment or pretreatment with 2% isoflurane induced HO-1 protein expression and caused an induction of HO activity. This result correlated with a decrease in iNOS expression, a decrease in the production of nitric oxide (NO) and impaired release of TNF-alpha in LPS stimulated macrophages. Blockade of HO activity with tin protoporphyrin (SnPP) reversed these effects. CONCLUSION: Isoflurane preconditioning exerts its anti inflammatory activity through the HO-1 pathway in an in vitro inflammation model. PMID- 19122673 TI - Notch1 signaling inhibits growth of EC109 esophageal carcinoma cells through downmodulation of HPV18 E6/E7 gene expression. AB - AIM: To investigate the role of the Notch1 signaling pathway in growth arrest of an esophageal carcinoma cell line (EC109) in vitro and the mechanism involved. METHODS: An intracellular domain of Notch1 (ICN) was transfected into cultured EC109 cells by lipofectamine transfection. Subsequently, the proliferation of the transfected cells was measured by an MTT assay. Cell cycle distribution was analyzed by flow cytometry. Human papillomavirus type 18 (HPV18) E6/E7 mRNA expression was detected by RT-PCR, and p53 protein expression was detected by Western blot. RESULTS: Activation of Notch1 signaling resulted in inhibition of EC109 cell proliferation with the induction of G(2)/M arrest, downmodulation of HPV18 E6/E7 gene expression, and upregulation of p53 expression. CONCLUSION: Repression of HPV18 E6/E7 expression by Notch1 signaling results in the activation of p53-mediated pathways with concomitant growth suppression of HPV18 positive EC109 cells. PMID- 19122674 TI - Deficiency of a beta-arrestin-2 signal complex contributes to insulin resistance. AB - Insulin resistance, a hallmark of type 2 diabetes, is a defect of insulin in stimulating insulin receptor signalling, which has become one of the most serious public health threats. Upon stimulation by insulin, insulin receptor recruits and phosphorylates insulin receptor substrate proteins, leading to activation of the phosphatidylinositol-3-OH kinase (PI(3)K)-Akt pathway. Activated Akt phosphorylates downstream kinases and transcription factors, thus mediating most of the metabolic actions of insulin. Beta-arrestins mediate biological functions of G-protein-coupled receptors by linking activated receptors with distinct sets of accessory and effecter proteins, thereby determining the specificity, efficiency and capacity of signals. Here we show that in diabetic mouse models, beta-arrestin-2 is severely downregulated. Knockdown of beta-arrestin-2 exacerbates insulin resistance, whereas administration of beta-arrestin-2 restores insulin sensitivity in mice. Further investigation reveals that insulin stimulates the formation of a new beta-arrestin-2 signal complex, in which beta arrestin-2 scaffolds Akt and Src to insulin receptor. Loss or dysfunction of beta arrestin-2 results in deficiency of this signal complex and disturbance of insulin signalling in vivo, thereby contributing to the development of insulin resistance and progression of type 2 diabetes. Our findings provide new insight into the molecular pathogenesis of insulin resistance, and implicate new preventive and therapeutic strategies against insulin resistance and type 2 diabetes. PMID- 19122675 TI - Ca(2+)/calmodulin regulates salicylic-acid-mediated plant immunity. AB - Intracellular calcium transients during plant-pathogen interactions are necessary early events leading to local and systemic acquired resistance. Salicylic acid, a critical messenger, is also required for both of these responses, but whether and how salicylic acid level is regulated by Ca(2+) signalling during plant-pathogen interaction is unclear. Here we report a mechanism connecting Ca(2+) signal to salicylic-acid-mediated immune response through calmodulin, AtSR1 (also known as CAMTA3), a Ca(2+)/calmodulin-binding transcription factor, and EDS1, an established regulator of salicylic acid level. Constitutive disease resistance and elevated levels of salicylic acid in loss-of-function alleles of Arabidopsis AtSR1 suggest that AtSR1 is a negative regulator of plant immunity. This was confirmed by epistasis analysis with mutants of compromised salicylic acid accumulation and disease resistance. We show that AtSR1 interacts with the promoter of EDS1 and represses its expression. Furthermore, Ca(2+)/calmodulin binding to AtSR1 is required for suppression of plant defence, indicating a direct role for Ca(2+)/calmodulin in regulating the function of AtSR1. These results reveal a previously unknown regulatory mechanism linking Ca(2+) signalling to salicylic acid level. PMID- 19122677 TI - Success of acupuncture treatment in patients with initially low sperm output is associated with a decrease in scrotal skin temperature. AB - Poor spermatogenesis in patients with inflammation of the genital tract is associated with scrotal hyperthermia. These patients can benefit from acupuncture treatment. We conducted a study to verify whether the influence of acupuncture treatment on sperm output in patients with low sperm density is associated with a decrease in scrotal temperature. The experimental group included 39 men who were referred for acupuncture owing to low sperm output. The control group, which comprised 18 normal fertile men, was used to define a threshold (30.5 degrees C) above which scrotal skin temperature was considered to be high. Accordingly, 34 of the 39 participants in the experimental group initially had high scrotal skin temperature; the other five had normal values. Scrotal skin temperature and sperm concentration were measured before and after acupuncture treatment. The five patients with initially normal scrotal temperatures were not affected by the acupuncture treatment. Following treatment, 17 of the 34 patients with hyperthermia, all of whom had genital tract inflammation, had normal scrotal skin temperature; in 15 of these 17 patients, sperm count was increased. In the remaining 17 men with scrotal hyperthermia, neither scrotal skin temperature nor sperm concentration was affected by the treatment. About 90% of the latter patients suffered from high gonadotropins or mixed etiological factors. Low sperm count in patients with inflammation of the genital tract seems to be associated with scrotal hyperthermia, and, consequently, acupuncture treatment is recommended for these men. PMID- 19122678 TI - Evaluation of erectile function after urethral reconstruction: a prospective study. AB - We conducted a prospective study of erectile dysfunction (ED) after urethral reconstructive surgery, using the 5-item International Index of Erectile Function (IIEF-5), the Sexual Life Quality Questionnaire (SLQQ) and the Quality of Life Questionnaire (QoLQ). Between January 2003 and July 2007, 125 male patients with urethral strictures underwent urethroplasty, and pre- and post-surgery erectile function was assessed using these three questionnaires. A formula to predict the probability of ED after urethroplasty was derived. At 3 months post-operatively, there was a significant decrease in IIEF-5 (16.57 +/- 7.98) and SLQQ scores (28.71 +/- 14.84) compared with pre-operative scores (P < 0.05). However, the IIEF-5 scores rebounded at 6 months post-operatively (17.22 +/- 8.41). Logistical regression analysis showed that the location of the urethral stricture, the recurrence of strictures and the choice of surgical technique were predictive of the post-operative occurrence of ED. This study identified the clinical risk factors for ED after urethroplasty. Posterior urethral stricture and end-to-end anastomosis were found to have a strong relationship with erectile function. The logistical model derived in this study may be applied to clinical decision algorithms for patients with urethral strictures. PMID- 19122676 TI - Mechanisms of regulated unconventional protein secretion. AB - Most eukaryotic proteins are secreted through the conventional endoplasmic reticulum (ER)-Golgi secretory pathway. However, cytoplasmic, nuclear and signal peptide-containing proteins have been shown to reach the cell surface by non conventional transport pathways. The mechanisms and molecular components of unconventional protein secretion are beginning to emerge, including a role for caspase 1 and for the peripheral Golgi protein GRASP, which could function as a plasma membrane tether for membrane compartments during specific stages of development. PMID- 19122679 TI - Tumor formation of prostate cancer cells influenced by stromal cells from the transitional or peripheral zones of the normal prostate. AB - This study was designed to investigate the different involvements of prostatic stromal cells from the normal transitional zone (TZ) or peripheral zone (PZ) in the carcinogenesis of prostate cancer (PCa) epithelial cells (PC-3) in vitro and in vivo co-culture models. Ultra-structures and gene expression profiles of primary cultures of human prostatic stromal cells from the normal TZ or PZ were analyzed by electron microscopy and microarray analysis. In vitro and in vivo co culture models composed of normal TZ or PZ stromal cells and human PCa PC-3 cells were established. We assessed tumor growth and weight in the in vivo nude mice model. There are morphological and ultra-structural differences in stromal cells from TZ and PZ of the normal prostate. In all, 514 differentially expressed genes were selected by microarray analysis; 483 genes were more highly expressed in stromal cells from TZ and 31 were more highly expressed in those from PZ. Co culture with PZ stromal cells and transforming growth factor-beta1 (TGF-beta1) increased the tumor growth of PC-3 cells in vitro and in vivo, as well as Bcl-2 expression. On the other hand, stromal cells of TZ suppressed PC-3 cell tumor growth in the mouse model. We conclude that ultra-structures and gene expression differ between the stromal cells from TZ or PZ of the normal prostate, and stroma epithelium interactions from TZ or PZ might be responsible for the distinct zonal localization of prostate tumor formation. PMID- 19122680 TI - Mammalian acatalasemia: the perspectives of bioinformatics and genetic toxicology. AB - The molecular defects in the catalase gene, levels of m-RNA and properties of the residual catalase studied by scientists are reviewed in human (Japanese, Swiss and Hungarian) and non-human (mouse and beagle dog) acatalasemia with reference to the bioinformatics. Japanese acatalasemia-I, the G to A transition at the fifth position of intron 4 of the catalase gene, limited the correct splicing of the mRNA and synthesized trace catalase with normal properties. Hungarian acatalasemia type C showed a splicing mutation. In the Japanese acatalasemia II and the type A and B of Hungarian acatalasemia, the deletion or insertion of nucleotides was observed in the coding regions, and the frame shift altered downstream amino acid sequences and formed truncated proteins. In the Hungarian acatalasemia D, the substitution of a nucleotide in the exon was found. In mouse and beagle dog acatalasemia, the substitution of nucleotides in the coding regions was also observed. Studies of residual catalase in Swiss, mouse and beagle dog acatalasemia showed that aberrant catalase protein degrades more quickly than normal catalase in cells. The experimental research in genetic toxicology concerning the effect of oxidative stressors (nitrogen monoxide, nitrogen dioxide and so on) on Japanese acatalasemic blood and acatalasemic mice is described. The clinical features of Japanese and Hungarian acatalasemic subjects are also described. PMID- 19122682 TI - A clinical investigation of the mechanism of loxoprofen, a non-steroidal anti inflammatory drug, for patients with nocturia. AB - We previously reported the effectiveness of loxoprofen sodium (loxoprofen), a non steroidal anti-inflammatory drug, for patients with lower urinary tract symptoms (LUTS) complaining of nocturia. In this study, we explored the mechanism of loxoprofen in the treatment of nocturia. Fifty-six patients complaining of nocturia were enrolled. They took a single 60-mg tablet of loxoprofen at bedtime for 14 days. The effects of this treatment were assessed by bladder diaries. Nocturia improved (nocturia decreased >or=1 void/night) in 40 patients (71.4%). Nocturnal urine volume was reduced in 31 of 40 (77.5%) without nocturnal single void volume increase. Nocturnal single-void volume increased in 4 of 40 (10.0%) without nocturnal urine volume reduction. Two of 40 (5.0%) demonstrated both nocturnal urine volume reduction and nocturnal single-void volume increase. Three (7.5%) were exceptions to the above. In conclusion, the main mechanism of loxoprofen is the reduction of nocturnal urine volume for the treatment of nocturia and the second mechanism is the increased bladder capacity. PMID- 19122681 TI - Toxin production by Aeromonas sobria in natural environments: river water vs. seawater. AB - Aeromonas are water-borne pathogens. They are halotolerant, which means that they can survive in environments whose salt content corresponds to that of seawater (3.0% NaCl). However, the presence of Aeromonas in seawater is extremely rare compared with that in river water. In this study, we tested the ability of Aeromonas sobria to produce toxins in river water and seawater. First, we cultured A. sobria on skim milk agar plates supplemented with either river water (SARW) or seawater (SASW). The bacteria grew on both plates. A clear zone around the bacteria was generated in SARW. However, such a zone was not observed in SASW, suggesting that proteases were not generated in SASW. Subsequently, we cultured A. sobria in a nutrient broth supplemented with either river water (NRW) or with seawater (NSW), and examined the protease activity of their culture supernatants. The protease activity of the culture supernatant from NSW was extremely low compared to that from NRW. The immunoblotting analysis showed that serine protease (ASP) was not produced by the culture in NSW. By contrast, aerolysin-like hemolysin was produced in all conditions examined in this study. This indicates that the salinity of water is deeply involved in the production of ASP by A. sobria. PMID- 19122683 TI - Initial report of hybrid radical prostatectomy for prostate cancer: reduced bleeding, clear vision, and secure surgical margins. AB - To evaluate morbidity in Hybrid Radical Prostatectomy (HRP, hybridized laparoscopic and open retropubic radical prostatectomy). The operative and pathological outcomes obtained in 25 consecutive patients who underwent HRP were reviewed. The median operating time was 220 min, median blood loss was 550 ml, and no patient required an allogenic blood transfusion. No severe postoperative complications were observed. The surgical margin was positive in 12% of all patients, and in 1 patient with pT2 or less (4.5%). These results indicate that HRP is safe and may be able to combine the benefits of both laparoscopic and open procedures. PMID- 19122684 TI - Dynamin 2 cooperates with amphiphysin 1 in phagocytosis in sertoli cells. AB - Testicular Sertoli cells highly express dynamin 2 and amphiphysin 1. Here we demonstrate that dynamin 2 is implicated in phosphatidylserine (PS)-dependent phagocytosis in Sertoli cells. Immunofluorescence and dual-live imaging revealed that dynamin 2 and amphiphysin 1 accumulate simultaneously at ruffles. These proteins are specifically bound in vitro. Over-expression of dominant negative dynamin 2 (K44A) inhibits liposome-uptake and leads to the mis-localization of amphiphysin 1. Thus, the cooperative function of dynamin 2 and amphiphysin 1 in PS-dependent phagocytosis is strongly suggested. PMID- 19122686 TI - The effect of N-acetylcysteine on biomarkers for radiation-induced oxidative damage in a rat model. AB - Our study aimed to investigate the potential radioprotective effects of N acetylcysteine (NAC) by comparing its biochemical effects with those of WR-2721, as a representative of clinically used radioprotectors, in preventing oxidative damage caused by gamma irradiation (single dose, 6Gy) in normal rat tissue. The rats (n=40) were divided randomly and equally into 4 groups:Control (C), Radiation (R), R+NAC (received irradiation and 1,000 mg/kg NAC) and R+WR-2721 (received irradiation and 200 mg/kg WR-2721) rats. Liver tissues and blood samples were harvested and utilized for reduced glutathione (GSH), malondialdehyde (MDA) and myeloperoxidase (MPO) detection. Serum and tissue GSH levels of R rats decreased compared to those of other groups (p<0.01). Tissue MDA levels of R+NAC and R+WR-2721 rats decreased compared to R rats (p<0.01; p<0.05, respectively). Tissue MPO activities of R+NAC and R+WR-2721 rats were higher than those of R rats (p<0.001). Serum MPO levels of R+WR-2721 rats were lower than those of C rats and R rats (p<0.01, p<0.001, respectively). In conclusion, the study suggests that the radioprotective effect against radiation-induced oxidative damage of NAC may be similar to that of WR-2721. PMID- 19122685 TI - Mechanistic analysis of resistance to REIC/Dkk-3-induced apoptosis in human bladder cancer cells. AB - We have recently shown that a new therapeutic modality using the REIC/Dkk-3 gene (Ad-REIC) is effective against various human cancers, including those of prostate, testis and breast origins. The aim of the present study was to examine the sensitivity of bladder cancers to Ad-REIC and to clarify the molecular mechanisms that determine sensitivity/resistance. We found that 2 human bladder cancer cell lines, T24 and J82, are resistant to Ad-REIC. In T24 and J82 cells, the ER stress response and activation of JNK were observed in a manner similar to that in the sensitive PC3 cells. Translocation of Bax to mitochondria occurred in PC3 cells but not in T24 and J82 cells. Bcl-2 was remarkably overexpressed in T24 and J82 compared with the expression levels in sensitive cell lines. Treatment of T24 and J82 cells with a Bcl-2 inhibitor sensitized the cells to Ad-REIC-induced apoptosis. The results indicate that some human bladder cancers are resistant to apoptosis induced by overexpression of REIC/Dkk-3, which is at least in part due to up-regulation of Bcl-2. These results provide a basis for possible use of Bcl 2 as a marker of sensitive cancers and to try to sensitize resistant cancers to Ad-REIC by down-regulation of Bcl-2. PMID- 19122687 TI - Abscess formation of the round ligament of the liver: report of a case. AB - Abscess formation of the round ligament of the liver is very rare. We report a case of a 70-year-old female with abscess of the round ligament after an endoscopic papillotomy for choledocholithiasis. On the 21st day following papillotomy, abscess formation of the round ligament was found by ultrasonographic examination. Surgical treatment was performed because conservative therapy was not effective. The purulent fluid and necrotic tissue at the round ligament were completely removed. Cultures obtained from the abscess grew Staphylococcus epidermidis, but the mechanism of abscess formation in this case remains unclear. PMID- 19122688 TI - Vaginoplasty with a pudendal-thigh flap in intersexuals. AB - We treated 2 different types of intersexual patients who underwent a vaginoplasty with the pudendal-thigh flap. One was a female with testicular feminization syndrome for whom we reconstructed the total vagina with a pudendal-thigh flap, and the other was a female with an adrenogenital syndrome for whom we enlarged the introitus of the vagina with the same approach. There were no complications such as a flap necrosis. In addition, there was no stricture of the neo-vagina and no urinary problem. PMID- 19122689 TI - Endoscopic mucosal resection of primary anorectal malignant melanoma: a case report. AB - Anorectal melanoma is a rare malignant tumor with a poor prognosis. However, several studies have reported cases of long-term survival. In this report, we present a patient with anorectal melanoma who has survived for 9 years after endoscopic mucosal resection. An 85-year-old man was referred to our hospital for further examination and treatment of an anal tumor 2 cm in size. Endoscopic ultrasonography revealed that the depth of tumor invasion was confined to the submucosal layer. Endoscopic mucosal resection was performed, and the tumor was diagnosed as a malignant melanoma. The patient was followed without any additional treatment, which was per his wishes. Although melanoma recurred 4 times thereafter, endoscopic mucosal resection was performed for each recurrent lesion. Thus, he has been alive for 9 years since the first endoscopic mucosal resection without distant metastases. If the depth of tumor invasion is shallow, endoscopic mucosal resection is a useful option among other therapeutic modalities. PMID- 19122690 TI - Time-resolved spectroscopy measurements of hydrogen-alpha, -beta, and -gamma emissions. AB - Hydrogen emission spectroscopy results are reported following laser-induced optical breakdown with infrared Nd:YAG laser radiation focused into a pulsed methane flow. Measurements of Stark-broadened atomic hydrogen-alpha, -beta, and gamma lines show electron number densities of 0.3 to 4x10(17) cm(-3) for time delays of 2.1 to 0.4 micros after laser-induced optical breakdown. In methane flow, recombination molecular spectra of the Delta nu = +2 progression of the C(2) Swan system are discernable in the H(beta) and H(gamma) plasma emissions within the first few microseconds. The recorded atomic spectra indicate the occurrence of hydrogen self-absorption for pulsed CH(4) flow pressures of 2.7x10(5) Pa (25 psig) and 6.5x10(5) Pa (80 psig). PMID- 19122691 TI - Detection sensitivity of laser-induced breakdown spectroscopy for Cr II in liquid samples. AB - The performance of laser-induced breakdown spectroscopy (LIBS) has been evaluated for detection of toxic metals such as Cr in water. Pure aqueous solutions (unitary matrix) with variable Cr concentration were used to construct calibration curves and to estimate the LIBS limit of detection (LOD). The calibration curves for Cr in a binary matrix (Cr plus Cd) and a tertiary matrix (Cr plus Cd and Co) were used to evaluate the matrix effect on the LOD. The LOD for Cr was found to be 1.1, 1.5, and 2.0 ppm (parts in 10(6)) in a unitary, binary, and tertiary matrix, respectively. Once calibrated, the system was utilized for the detection and quantification of the Cr in tannery wastewater collected from different locations in the industrial area of Kanpur, India, where Cr concentrations were determined to be far higher than the U.S. Environmental Protection Agency safe drinking water limit of 0.05 ppm. PMID- 19122692 TI - Multivariate analysis of standoff laser-induced breakdown spectroscopy spectra for classification of explosive-containing residues. AB - A technique being evaluated for standoff explosives detection is laser-induced breakdown spectroscopy (LIBS). LIBS is a real-time sensor technology that uses components that can be configured into a ruggedized standoff instrument. The U.S. Army Research Laboratory has been coupling standoff LIBS spectra with chemometrics for several years now in order to discriminate between explosives and nonexplosives. We have investigated the use of partial least squares discriminant analysis (PLS-DA) for explosives detection. We have extended our study of PLS-DA to more complex sample types, including binary mixtures, different types of explosives, and samples not included in the model. We demonstrate the importance of building the PLS-DA model by iteratively testing it against sample test sets. Independent test sets are used to test the robustness of the final model. PMID- 19122693 TI - Single pulse laser-induced breakdown spectroscopy of bulk aqueous solutions at oceanic pressures: interrelationship of gate delay and pulse energy. AB - The ability of oceanographers to make sustained measurements of ocean processes is limited by the number of available sensors for long-term in situ analysis. In recent years, laser-induced breakdown spectroscopy (LIBS) has been identified as a viable technique to develop into an oceanic chemical sensor. We performed single pulse laser-induced breakdown spectroscopy of high pressure bulk aqueous solutions to detect three analytes (sodium, manganese, and calcium) that are of key importance in hydrothermal vent fluids, an ocean environment that would greatly benefit from the development of an oceanic LIBS sensor. The interrelationship of the key experimental parameters, pulse energy and gate delay, for a range of pressures up to 2.76x10(7) Pa, is studied. A minimal effect of pressure on the peak intensity is observed. A short gate delay (less than 200 ns) must be used at all pressures. The ability to use a relatively low laser pulse energy (less than approximately 60 mJ) for detection of analytes at high pressure is also established. Na, Mn, and Ca are detectable at pressures up to 2.76x10(7) Pa at 50, 500, and 50 ppm, respectively, using an Echelle spectrometer. PMID- 19122694 TI - Double pulse laser-induced breakdown spectroscopy of bulk aqueous solutions at oceanic pressures: interrelationship of gate delay, pulse energies, interpulse delay, and pressure. AB - Laser-induced breakdown spectroscopy (LIBS) has been identified as an analytical chemistry technique suitable for field use. We use double pulse LIBS to detect five analytes (sodium, manganese, calcium, magnesium, and potassium) that are of key importance in understanding the chemistry of deep ocean hydrothermal vent fluids as well as mixtures of vent fluids and seawater. The high pressure aqueous environment of the deep ocean is simulated in the laboratory, and the key double pulse experimental parameters (laser pulse energies, gate delay time, and interpulse delay time) are studied at pressures up to 2.76x10(7) Pa. Each element is found to have a unique optimal set of parameters for detection. For all pressures and energies, a short (< or = 100 ns) gate delay is necessary. As pressure increases, a shorter interpulse delay is needed and the double pulse conditions effectively become single pulse for both the 1.38x10(7) Pa and the 2.76x10(7) Pa conditions tested. Calibration curves reveal the limits of detection of the elements (5000 ppm Mg, 500 ppm K, 500 ppm Ca, 1000 ppm Mn, and 50 ppm Na) in aqueous solutions at 2.76x10(7) Pa for the experimental setup used. When compared to our previous single pulse LIBS work for Ca, Mn, and Na, the use of double pulse LIBS for analyte detection in high pressure aqueous solutions did not improve the limits of detection. PMID- 19122696 TI - Comparison of laser-induced breakdown spectra of organic compounds with irradiation at 1.5 and 1.064 microm. AB - A comprehensive investigation of laser-induced breakdown spectroscopy (LIBS) at 1.500 microm of residues of six organic compounds (anthracene, caffeine, glucose, 1,3-dinitrobenzene, 2,4-dinitrophenol, and 2,4-dinitrotoluene) on aluminum substrates is presented and compared with LIBS at the Nd:YAG fundamental wavelength of 1.064 microm. The overall emission intensities were found to be smaller at 1.500 microm than at 1.064 microm, and the ratios of C(2) and CN molecular emissions to the H atomic emissions were observed to be less. Possible reasons for the observed differences in LIBS at 1.064 microm versus 1.500 microm are discussed. PMID- 19122695 TI - Laser-induced breakdown spectroscopy measurement in methane and biodiesel flames using an ungated detector. AB - Laser-induced breakdown spectroscopy (LIBS) has been applied to measure the equivalence ratio of CH(4)/air flames using gated detection. In this work, we have developed an ungated, miniature LIBS-based sensor for studying CH(4)/air and biodiesel flames. We have used this sensor to characterize the biodiesel flame. LIBS spectra of biodiesel flames were recorded with different ethanol concentrations in the biodiesel and also at different axial locations within the flame. The sensor performance was evaluated with a CH(4)/air flame. LIBS signals of N, O, and H from a CH(4)/air flame were used to determine the equivalence ratio. A linear relationship between the intensity ratio of H and O lines and the calculated equivalence ratio were obtained with this sensor. PMID- 19122697 TI - Chemical taggant detection and analysis by laser-induced breakdown spectroscopy. AB - A commercially available chemical identification taggant that imparts a unique elemental fingerprint to any object and can be analytically distinguished from billions of possible combinations has been developed. The liquid tag is easily applied and, once dry, can be removed and analyzed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to determine the combination of elements present in the sample. The current study investigates the use of laser induced breakdown spectroscopy (LIBS) as an alternative, and perhaps more practical, analysis scheme to LA-ICP-MS for this taggant. LIBS provides excellent discrimination potential, sensitivity, and repeatability of analysis for up to 17 rare-earth elements using a Nd:YAG 266 nm or 1064 nm laser and an intensified CCD detector. PMID- 19122698 TI - Extraction of information from laser-induced breakdown spectroscopy spectral data by multivariate analysis. AB - Laser-induced breakdown spectroscopy (LIBS) is being proposed more and more as a high-throughput technology to assess the elemental composition of materials. When a specific element is of interest, semiquantification is possible by building a calibration model using the emission line intensity of this element for known samples. However, a unique element has usually more than one emission line, and there are many examples where several emission lines used in combination give dramatically better results than any of the individual variables used alone. With a multivariate approach, models can be constructed that take into account all the emission lines related to a specific element; therefore more robust models can be developed. In this work, chemometric methods such as principal component analysis and partial least squares are proposed to resolve and extract useful information from the LIBS spectral data collected on biological materials. PMID- 19122699 TI - Time-dependent single and double pulse laser-induced breakdown spectroscopy of chromium in liquid. AB - A study of aqueous solutions of chromium using single and double pulse laser induced breakdown spectroscopy (LIBS) is presented. Three atomic emission lines show enhancement in emission under dual pulse LIBS excitation. The temporal evolution of line emission indicates that a shock wave front produced by the first laser pulse plays an important role in determining the decay rate of intensity by excitation transfer in single pulse LIBS and by plasma confinement in double pulse LIBS. The ratio of emission in dual pulse LIBS to single pulse LIBS with time shows a linear increase followed by the onset of saturation. A theoretical calculation of the enhancement is found to be in qualitative agreement with the experimental results, suggesting that material ablation in dual pulse LIBS should be > or = 3.5 times that of single pulse LIBS. There is indication that the increase in ablation and subsequent enhancement in emission may be due to the rarefied gas density inside the region enclosed by the shock wave produced by the first laser pulse. The limit of detection of Cr in aqueous solution has been improved by an order of magnitude with double pulse LIBS. PMID- 19122700 TI - Theoretical model for double pulse laser-induced breakdown spectroscopy. AB - We present a simple theoretical model for the emission from double pulse laser induced plasmas that was developed to better understand the processes and factors involved in enhancement of plasma emission. In this model, the plasma emission is directly proportional to the square of plasma density, its volume, and the fraction of second laser pulse absorbed through inverse bremsstrahlung absorption by the plasma plume of the first laser pulse. The electron-ion collision frequency determines the profile and location of the peak of emission enhancement with respect to the delay between the two lasers, whereas the amplitude of the enhancement is mainly dependent on the increase in the mass ablation rate after the second laser pulse. The effects of increase in temperature and in plasma volume after the second laser pulse are also discussed in light of this model. PMID- 19122702 TI - Detection of indoor biological hazards using the man-portable laser induced breakdown spectrometer. AB - The performance of a man-portable laser induced breakdown spectrometer was evaluated for the detection of biological powders on indoor office surfaces and wipe materials. Identification of pure unknown powders was performed by comparing against a library of spectra containing biological agent surrogates and confusant materials, such as dusts, diesel soot, natural and artificial sweeteners, and drink powders, using linear correlation analysis. Simple models constructed using a second technique, partial least squares discriminant analysis, successfully identified Bacillus subtilis (BG) spores on wipe materials and office surfaces. Furthermore, these models were able to identify BG on materials not used in the training of the model. PMID- 19122701 TI - Quantitative analysis of gallstones using laser-induced breakdown spectroscopy. AB - The utility of laser-induced breakdown spectroscopy (LIBS) for categorizing different types of gallbladder stone has been demonstrated by analyzing their major and minor constituents. LIBS spectra of three types of gallstone have been recorded in the 200-900 nm spectral region. Calcium is found to be the major element in all types of gallbladder stone. The spectrophotometric method has been used to classify the stones. A calibration-free LIBS method has been used for the quantitative analysis of metal elements, and the results have been compared with those obtained from inductively coupled plasma atomic emission spectroscopy (ICP AES) measurements. The single-shot LIBS spectra from different points on the cross section (in steps of 0.5 mm from one end to the other) of gallstones have also been recorded to study the variation of constituents from the center to the surface. The presence of different metal elements and their possible role in gallstone formation is discussed. PMID- 19122703 TI - Determination of thorium and uranium in solution by laser-induced breakdown spectrometry. AB - Laser-induced breakdown spectrometry (LIBS) has been successfully developed to quantify thorium (Th) and uranium (U) in solution using membrane-based filter paper as a sample support. The filter paper is easy to handle and the entire analysis process takes only a few minutes for each sample. The developed method provides a practical approach for fast and sensitive determination of actinides in aqueous solutions using LIBS. We obtained calibration curves for U and Th individually as well as as a mixture. We observed that the quantification of trace levels of U in a bulk amount of Th was not possible because of strong spectral interference from Th at the most intense lines of U, but traces of Th in a bulk amount of U could be determined. The concentrations of U and Th in unknown solutions were determined by use of LIBS, and these results agreed to within 4% and 2%, respectively, with the expected values. The limits of detection for Th and U were calculated from the experimental data and were in the range of a few parts per million by weight to a few tens of parts per million by weight. PMID- 19122704 TI - Dynamics of laser-ablated carbon plasma: formation of C2 and CN. AB - We report time-resolved imaging of a laser-ablated carbon plasma plume to investigate the expansion dynamics of C(2) and CN in an ambient atmosphere of nitrogen gas at various pressures. An attempt is made to locate C(2) and CN species in the carbon plasma plume and correlate them with the results of spectroscopic observations. The ablated C(2) and CN species decelerate due to collisions with nitrogen gas and are localized in the slower part (approximately 300 ns) of the expanding plume. Further expansion (<700 ns) of the plasma reveals the concentration of C(2) species on the periphery of the plume, whereas CN dominates at the core of the plume. However, at times greater than 700 ns, the collisions and recombination processes dominate in the plume and C(2) expands slower than CN. The plume dynamics is studied in terms of shock-wave and drag models. PMID- 19122705 TI - Commercialization of laser-induced breakdown spectroscopy for lead-in-paint inspection. AB - A study was undertaken to determine if laser-induced breakdown spectroscopy (LIBS) can be a practical and competitive alternative to x-ray fluorescence (XRF) methods for lead-in-paint inspection. Experiments in the laboratory confirmed that LIBS is suitable for detecting lead in paint at the hazard levels defined by federal agencies. Although we compared speed, function, and cost, fundamental differences between the XRF and LIBS measurements limited our ability to make a quantitative performance comparison. While the LIBS method can achieve the required sensitivity and offers a way to obtain unique information during inspection, the current component costs will likely restrict interest in the method to niche applications. PMID- 19122706 TI - Use of laser induced breakdown spectroscopy in the determination of gem provenance: beryls. AB - The provenance of gem stones has been of interest to geologists, gemologists, archeologists, and historians for centuries. Laser induced breakdown spectroscopy (LIBS) provides a minimally destructive tool for recording the rich chemical signatures of gem beryls (aquamarine, goshenite, heliodor, and morganite). Broadband LIBS spectra of 39 beryl (Be(3)Al(2)Si(6)O(18)) specimens from 11 pegmatite mines in New Hampshire, Connecticut, and Maine (USA) are used to assess the potential of using principal component analysis of LIBS spectra to determine specimen provenance. Using this technique, beryls from the three beryl-bearing zones in the Palermo #1 pegmatite (New Hampshire) can be recognized. However, the compositional variation within this single mine is comparable to that in beryls from all three states. Thus, a very large database with detailed location metadata will be required to routinely determine gem beryl provenance. PMID- 19122707 TI - Laser-induced breakdown spectroscopy for the classification of unknown powders. AB - Laser-induced breakdown spectroscopy (LIBS) was used to discern between two biological agent surrogates (Bacillus atrophaeus and ovalbumin) and potential interferent compounds (mold spores, humic acid, house dust, and Arizona road dust). Multiple linear regression and neural network analysis models were constructed by using B. atrophaeus and ovalbumin spectra, and limits of detection were calculated. Classification of the agent surrogates' LIBS spectra was attempted by using a neural network model. False negative rates of 0% were observed for B. atrophaeus (100 colony forming units) spore spectra with the neural network model used for classification. PMID- 19122708 TI - Movable fiber probe for gas-phase laser-induced breakdown spectroscopy. AB - A movable probe that fiber couples both the beam delivery and the signal collection functions of gas-phase laser-induced breakdown spectroscopy (LIBS) measurements was evaluated. The adjustable probe was used to investigate the effect of delivery fiber curvature on plasma characteristics and the associated effect on LIBS spectra and to further identify issues remaining to facilitate fully fiber-coupled gas-phase LIBS measurements. LIBS data were collected from lean methane-air mixtures of various equivalence ratios and spectroscopically analyzed to establish the ability to determine relative fuel-air ratio. Measurements with straight delivery fiber were compared to those with the fiber curved at specific radii. Decreasing fiber radius of curvature decreased fiber transmission efficiency and reduced the spark formation probability by almost a factor of 2. For constant fiber input energy, this decreased transmission increased the percentage of failed spark formations and influenced the LIBS elemental ratio calculations. However, minimal difference was found between LIBS measurements with straight or curved fiber as long as the output energy and a constant laser beam spot diameter were maintained on the exit beam focusing lens. A significant reduction in data scatter and improved linearity were achieved by using the Balmer series H(alpha) and H(beta) hydrogen emission line ratio as a data selection criterion. Observed linear variation of H/N elemental ratio with equivalence ratio confirmed the possibility of a flexible, light-contained, fully fiber-coupled probe for remote gas-phase LIBS analysis. PMID- 19122709 TI - Elemental analysis of laser induced breakdown spectroscopy aided by an empirical spectral database. AB - Laser induced breakdown spectroscopy (LIBS) is commonly used to identify elemental compositions of various samples. To facilitate this task, we propose the use of an elemental spectral library for single-pulsed, nanosecond LIBS in the spectral range 198-968 nm. This spectroscopic library is generated by measuring optical emissions from plasmas of 40 pure elements. To demonstrate the usefulness of the proposed database, we measure and analyze the LIBS spectra of pure iron and of ethanol and show that we identify these samples with a high degree of certainty. PMID- 19122711 TI - High-irradiance reactors with unfolded aplanatic optics. AB - Reconstituting the intense irradiance of short-arc discharge lamps at a remote target, at high radiative efficiency, represents a central challenge in the design of high-temperature chemical reactors, heightened by the need for high numerical aperture at both the target and the source. Separating the optical system from both the source and the reactor allows pragmatic operation, monitoring, and control. We explore near-field unfolded aplanats as feasible solutions and report measurements for a prototype that constitutes a double ellipsoid mirror. We also propose compound unfolded aplanats that collect lamp emission over all angles (in lieu of light recycling optics) and irradiate the reactor over nearly its full circumference. PMID- 19122712 TI - Modal filtering for midinfrared nulling interferometry using single mode silver halide fibers. AB - We demonstrate the modal filtering properties of newly developed single mode silver halide fibers for use at midinfrared wavelengths, centered at 10.5 microm. The goal was to achieve a suppression of nonfundamental modes greater than a factor of 300 to enable the detection and characterization of Earthlike exoplanets with a space-based nulling interferometer. Fiber segments of 4.5 cm, 10.5 cm, 15 cm, and 20 cm lengths were tested. We find that the performance of the fiber was limited not by the modal filtering properties of the core but by the unsuppressed cladding modes present at the output of the fiber. In 10.5 cm and longer sections, this effect can be alleviated by properly aperturing the output. Exclusive of coupling losses, the fiber segments of 10.5-20 cm length can provide power suppression of undesirable components of the input field by a factor of 15,000 at least. The demonstrated performance thus far surpasses our requirements, such that even very short sections of fiber provide adequate modal filtering for exoplanet characterization. PMID- 19122713 TI - Measurement and simulation of the effect of snowfall on free-space optical propagation. AB - We measured the time variation of a received laser signal level during snowfall over a distance of 72 m. The signal level dropped sharply for up to 10 ms when a snowflake crossed the laser beam. The probability distribution of the variation due to snowfall was calculated by assuming it to be the linear superposition of the light diffracted by snowflakes. The measured distributions could be reproduced by assuming reasonable snowflake size distributions. Furthermore, the probability distributions due to snowfall over a 1 km distance were calculated, and the expected bit errors during snowfall and the transmitted beam sizes were evaluated. PMID- 19122714 TI - Observing the solar corona with a tunable Fabry-Perot filter. AB - A solid Fabry-Perot etalon with a 0.16 A passband was used during the 180 s solar eclipse of 2006 for rapid scans of an emission line of the solar corona. The etalon was a Y-cut lithium niobate wafer coated with reflective and conductive (ITO) layers. Voltage applied perpendicular to the etalon face produced a passband shift of 0.0011 A V(-1). During the eclipse, 18 filtergrams were obtained at six 0.22 A steps across the profile of the forbidden [Fe X] spectral emission line at 6374.4 A, which results from the 10(6) K coronal plasma. The 9.3 x 9.3 arcmin field of view showed the structure of the corona above a newly emerged sunspot region. We discuss tests performed on the etalon before and after the eclipse. We also discuss the coronal observations, which show some features with 10 km s(-1) velocities in the line of sight. PMID- 19122715 TI - Planar waveguide obtained by burying a Ge22As(20Se58 fiber in As2S3 glass. AB - We demonstrate the possibility of fabricating an infrared transmitting waveguide by burying fiber in chalcogenide glasses. Two highly mature chalcogenide glasses are used for these experiments. GASIR glass from Umicore IR Glass, Olen, Belgium, with the composition of Ge(22)As(20)Se(58) is used to draw fibers that are then buried in an As(2)S(3) glass substrate. The glasses we used are compatible, and we obtained a high quality interface. We performed a transmission test with a CO(2) laser at 9.3 microm. The potential for extremely low loss planar waveguides is discussed. PMID- 19122716 TI - X-ray modulation transfer functions of photostimulable phosphor image plates and scanners. AB - The modulation transfer functions of two types of photostimulable phosphor image plates were determined in the 10 keV to 50 keV x-ray energy range using a resolution test pattern with up to 10 line pairs per mm (LP/mm) and a wavelength dispersive x-ray spectrometer. Techniques were developed for correcting for the partial transmittance of the high energy x rays through the lead bars of the resolution test pattern, and the modulation transfer function (MTF) was determined from the measured change in contrast with LP/mm values. The MTF was convolved with the slit function of the image plate scanner, and the resulting point spread functions (PSFs) were in good agreement with the observed shapes and widths of x-ray spectral lines and with the PSF derived from edge spread functions. The shapes and the full width at half-maximum (FWHM) values of the PSF curves of the Fuji Superior Resolution (SR) and Fuji Maximum Sensitivity (MS) image plate detectors, consisting of the image plate and the scanner, determined by the three methods gave consistent results: The SR PSF is Gaussian with 0.13 mm FWHM, and the MS PSF is Lorentzian with 0.19 mm FWHM. These techniques result in the accurate determination of the spatial resolution achievable using image plate and scanner combinations and enable the optimization of spatial resolution for x ray spectroscopy and radiography. PMID- 19122717 TI - Broadly tunable ultraviolet light generation in a compact MgO-doped periodically poled stoichiometric lithium tantalate optical parametric oscillator with a high Q cavity. AB - A compact tunable UV laser source based on intracavity sum frequency generation in a MgO-doped periodically-poled stoichiometric lithium tantalate optical parametric oscillator is reported. UV output at an approximately 1 mW level of power over the range of 364 to 378 nm with a bandwidth of 0.5 nm was obtained with a crystal that has just one periodically-poled grating period. The full tuning range can be as much as approximately 68 nm, from 324 to 392 nm by varying the crystal temperature from room temperature to 250 degrees C. This will cover nearly the entire UVA range. PMID- 19122718 TI - Detection of condensed-phase explosives via laser-induced vaporization, photodissociation, and resonant excitation. AB - We investigate the remote detection of explosives via a technique that vaporizes and photodissociates the condensed-phase material and detects the resulting vibrationally excited NO fragments via laser-induced fluorescence. The technique utilizes a single 7 ns pulse of a tunable laser near 236.2 nm to perform these multiple processes. The resulting blue-shifted fluorescence (226 nm) is detected using a photomultiplier and narrowband filter that strongly block the scatter of the pump laser off the solid media while passing the shorter wavelength photons. Various nitro-bearing compounds, including 2,6-dinitrotoluene (DNT), 2,4,6 trinitrotoluene (TNT), pentaerythritol tetranitrate (PETN), and hexahydro-1,3,5 trinitro-1,3,5-triazine (RDX) were detected with a signal-to-noise of 25 dB. The effects of laser fluence, wavelength, and sample morphology were examined. PMID- 19122719 TI - Observation of nonlinear transmission enhancement in cavities filled with nonlinear organic materials. AB - We show experimental and theoretical results for enhancement of nonlinear transmission (NT) in moderate finesse cavities filled with nonlinear organic materials (NLOM). Our design for enhancement of nonlinear transmission using micro NLOM cavities compared with reference samples of the same material show that single cavities can enhance the nonlinear response by a factor of 10 or greater under high-absorption conditions. Further enhancement can be achieved in multiple-cavity structures. Other advantages of the cavity structures for nonlinear transmission, such as a higher damage threshold and a broader NT band, are also discussed. Our initial experimental results show a threefold reduction in the nonlinear threshold fluence in a single cavity device compared directly to an identical sample without mirrors, in qualitative agreement with our calculations. PMID- 19122720 TI - Acceleration method of computing a compensated phase-added stereogram on a graphic processing unit. AB - We have implemented experimental code to compute a compensated phase-added stereogram (CPAS), which was proposed in a previous paper, on a graphic processing unit (GPU). In this paper, we show an acceleration method for CPAS computation by means of the GPU and compare the computation time between CPU based and GPU-based calculations, which are programmed in our laboratories. In addition, we demonstrate their reconstructed images. As a result, we could achieve a performance gain of a factor of over 33 compared with a CPU-based computing environment and digital holograms can be displayed at 30 frames per second with 15,000 points. PMID- 19122721 TI - Chemical imaging with Fourier transform coherent anti-Stokes Raman scattering microscopy. AB - We report chemical imaging using Fourier transform coherent anti-Stokes Raman scattering (FTCARS) microscopy. Adding a passively phase-stable local field to amplify the weak FTCARS signal, we also demonstrate interferometric FTCARS microscopy, permitting reduced incident power to be used for imaging. We discuss signal-to-noise considerations and the conditions necessary to effectively suppress background noise, allowing FTCARS microscopy that is limited by the shot noise of the detector. We also discuss differences between the signal-to-noise obtainable by time and frequency domain coherent anti-Stokes Raman scattering (CARS) methods. PMID- 19122722 TI - Thermal infrared radiance simulation with aggregation modeling (TITAN): an infrared radiative transfer model for heterogeneous three-dimensional surface- application over urban areas. AB - The thermal infrared radiance simulation with aggregation modeling (TITAN) model, presented here, is an innovative transfer radiative code in the infrared domain (3-14 microm). It takes into account the three-dimensional (3D) structure of the landscape and simulates all the radiative components introduced by this 3D structure, which are due to the reflection and emission of walls and sloping roofs. Examples are given to illustrate the new opportunities offered by TITAN over urban areas. First, a phenomenological study is conducted at four wavelengths analyzing the relative effect of all the radiative contributors to the total signal. The same analysis is performed at bottom of atmosphere, which reveals an error occurring when a flat assumption is made (between 1% and 5%). In a second example, the directional effects at sensor level are simulated and show that the radiative temperature can vary by up to 10 K. PMID- 19122723 TI - Analysis of simulated and experimental backscattered images of turbid media in linearly polarized light: estimation of the anisotropy factor. AB - Optical characterization of biological tissues is of real interest to improve medical diagnosis, in particular in the detection of precancerous tissues. We propose a new, noninvasive method allowing the estimation of the anisotropy factor. This method is based on the image analysis of the Q element of the Stokes vector backscattered from the turbid medium. These Q-element images show specific patterns depending on g. Therefore the use of Fourier descriptors (FDs) on simulated data to discriminate the specific geometrical features of the Q element enabled us to determine a linear relation between the anisotropy factor and six FDs. This method was applied on experimental data obtained with calibrated solutions. The anisotropy factor was estimated with a maximum relative error of 13%. PMID- 19122724 TI - Development of a switched integrator amplifier for high-accuracy optical measurements. AB - In the field of low flux optical measurements, the development and use of large area silicon detectors is becoming more frequent. The current/voltage conversion of their photocurrent presents a set of problems for traditional transimpedance amplifiers. The switched integration principle overcomes these limitations. We describe the development of a fully characterized current-voltage amplifier using the switched integrator technique. Two distinct systems have been developed in parallel at the United Kingdom's National Physical Laboratory (NPL) and Czech Metrology Institute (CMI) laboratories. We present the circuit theory and best practice in the design and construction of switched integrators. In conclusion the results achieved and future developments are discussed. PMID- 19122725 TI - Evaluation of expanded uncertainties in luminous intensity and illuminance calibrations. AB - Detector-based calibrating methods and expressions for calculation of photometric uncertainties related to uncertainties in the calibrations of luminous intensity of a light source, illuminance responsivity of a photometer head, and calibration factors of an illuminance meter are discussed. These methods permit luminous intensity calibrations of incandescent light sources, luminous responsivity calibrations of photometer heads, and calibration factors of illuminance meters to be carried out with relative expanded uncertainties (with a level of confidence of 95.45%) of 0.4%, 0.4%, and 0.6%, respectively. PMID- 19122726 TI - Double optical limiting in gold nanoshell: tuning from visible to infrared region by shell thickness. AB - Because of surface plasmon resonance (SPR) induced nonlinear absorption, there are two optical limiting bands in gold nanoshells. The longer and shorter wavelength optical limiting bands correspond to the symmetric and antisymmetric coupling resonance absorption modes, respectively. Theoretical calculations based on quasi-static approximation show that the longer wavelength optical limiting band red shifts from the visible to the infrared region by decreasing the shell thickness. A mechanism based on polarization direction of a local electric field in a gold shell is investigated to describe the wavelength shift of the optical limiting band. PMID- 19122728 TI - Free-electron-laser-induced shock-wave control and mechanistic analysis using pulse control. AB - The wavelength of the free electron laser (FEL) in Osaka University can be continuously varied in the range of 5.0-20.0 microm. The FEL has a double-pulse structure, consisting of a train of macropulses of pulse duration 12 micros. Each macropulse contains a train of 330 micropulses of pulse duration 5 ps. The tunability and picosecond pulses afford new medical and biological applications. However, a macropulse of long pulse duration leads to undesirable secondary effects. Precise control of the macropulse duration is essential for the high precision applications of the FEL. An FEL pulse control system using acousto optic modulators has been developed to investigate mechanical (shock-wave) effects of the FEL on living tissues. With this system, we have controlled photoinduced shock waves and determine the mechanism of interaction during FEL induced tissue ablation. PMID- 19122729 TI - Development of a two-line OH-laser-induced fluorescence thermometry diagnostics strategy for gas-phase temperature measurements in engines. AB - This study aims at optimizing two-line OH thermometry strategies for in-cylinder measurement in internal combustion engines. Various aspects are investigated experimentally, such as the selection of suitable OH lines and the possibility of using a single calibration coefficient for variable mixture composition, temperature, and pressure conditions. Two kinds of experimental systems have been investigated. First, a laminar methane-air burner flame at atmospheric pressure, whose stability allowed the determination of OH-laser-induced fluorescence (LIF) intensity ratios from nonsimultaneous imaging. The temperature distribution in the flame is presented for OH-transition pairs with various temperature sensitivities. The burner flame was studied for equivalence ratios from phi=0.93 to 1.30 in order to check for the stability of calibration over various flame conditions. Additionally, OH LIF images were acquired in an optical engine for the chosen OH transitions yielding data about the effect of pressure on OH LIF signals under realistic experimental conditions. PMID- 19122730 TI - Effect of surface roughness on the extinction-based localized surface plasmon resonance biosensors. AB - We investigated the effect of surface roughness on the sensor performance of extinction-based localized surface plasmon resonance (LSPR) biosensors. The sensor measures resonance wavelength shifts in transmittance caused by biomolecular interactions that are amplified by periodic nanostructures. The numerical computation was conducted using rigorous coupled-wave analysis with Gaussian random surface profiles. The results suggest that, when a surface has a roughness smaller than 2 nm in height deviation, the sensitivity of a LSPR biosensor is not significantly influenced regardless of correlation length (CL). However, we found that the extinction peak amplitude and curve width are affected substantially by a decrease in CL. At a less than 100 nm CL, surface roughness can induce interference between localized surface plasmons excited by the surface and nanowires, which can lead to major degradation of sensor performance. PMID- 19122731 TI - Defocus sensitivity optimization using the defocus Taylor expansion of the optical transfer function. AB - The Taylor expansion of the incoherent optical transfer function with respect to defocus is a valuable tool in the design and analysis of computational imaging systems. It efficiently describes the behavior of the system near best focus and beyond. Formulas for computing the coefficients in this expansion are derived and shown to be amenable to efficient digital calculation. Their application to the design of phase masks for systems insensitive to defocus aberrations and for systems that estimate object range are explored. PMID- 19122732 TI - Multichanneled encryption via a joint transform correlator architecture. AB - We propose a multichanneling encryption method by using multiple random-phase mask apertures in the input plane based on a joint transform correlation scheme. In the proposal, this multiple aperture arrangement is changed as different input objects are inserted and stored. Then, during the decryption step, the appropriate use of the random-phase mask apertures can ensure the retrieval of different information. This approach provides different access levels. Computer simulations show the potential of the technique and experimental results verify the feasibility of this method. PMID- 19122727 TI - In-vacuum optical isolation changes by heating in a Faraday isolator. AB - We describe a model evaluating changes in the optical isolation of a Faraday isolator when passing from air to vacuum in terms of different thermal effects in the crystal. The changes are particularly significant in the crystal thermal lensing (refraction index and thermal expansion) and in its Verdet constant and can be ascribed to the less efficient convection cooling of the magneto-optic crystal of the Faraday isolator. An isolation decrease by a factor of 10 is experimentally observed in a Faraday isolator that is used in a gravitational wave experiment (Virgo) with a 10 W input laser when going from air to vacuum. A finite element model simulation reproduces with a great accuracy the experimental data measured on Virgo and on a test bench. A first set of measurements of the thermal lensing has been used to characterize the losses of the crystal, which depend on the sample. The isolation factor measured on Virgo confirms the simulation model and the absorption losses of 0.0016 +/- 0.0002/cm for the TGG magneto-optic crystal used in the Faraday isolator. PMID- 19122733 TI - Coupling method for laser-diode-array end-pumped solid-state amplifiers. AB - A new coupling method is proposed for high-power laser-diode-array (LDA) end pumped large-aperture amplifiers. In this method, stacks in the LDA are distributed on a spherical surface, and a hollow duct is placed after them. Both numerical simulations and experimental research have been done for this method. All results show that high coupling efficiency and uniform pumping field can be obtained simultaneously using this method. PMID- 19122734 TI - Straight multimode interference phased array structure using periodic segmented waveguide phase array. AB - A new design of a multimode interference (MMI) phased array structure is proposed. This design is based on replacing the arrayed delay arms between the first MMI power splitter and the second MMI combiner by periodic segmented waveguides. This allows a straight structure without curved waveguides and thus reduces greatly the size of the structure. A design example of an 8 channel multiplexer is presented showing a size reduction by a factor of 23 when compared with conventional design, while keeping nearly the same performance. PMID- 19122735 TI - Radiative transfer model for aerosols at infrared wavelengths for passive remote sensing applications: revisited. AB - We introduced a two-dimensional radiative transfer model for aerosols in the thermal infrared [Appl. Opt.45, 6860-6875 (2006)APOPAI0003 693510.1364/AO.45.006860]. In that paper we superimposed two orthogonal plane parallel layers to compute the radiance due to a two-dimensional (2D) rectangular aerosol cloud. In this paper we revisit the model and correct an error in the interaction of the two layers. We derive new expressions relating to the signal content of the radiance from an aerosol cloud based on the concept of five directional thermal contrasts: four for the 2D diffuse radiance and one for direct radiance along the line of sight. The new expressions give additional insight on the radiative transfer processes within the cloud. Simulations for Bacillus subtilis var. niger (BG) bioaerosol and dustlike kaolin aerosol clouds are compared and contrasted for two geometries: an airborne sensor looking down and a ground-based sensor looking up. Simulation results suggest that aerosol cloud detection from an airborne platform may be more challenging than for a ground-based sensor and that the detection of an aerosol cloud in emission mode (negative direct thermal contrast) is not the same as the detection of an aerosol cloud in absorption mode (positive direct thermal contrast). PMID- 19122736 TI - Automated collimation testing using a temporal phase shifting technique in Talbot interferometry. AB - We propose a quantitative approach toward the detection of collimation of a coherent optical beam. For detection of collimation position of the collimator, a temporal phase shifting technique has been incorporated into Talbot interferometry. The slope of the phase maps depicts the collimation position of the collimator. This technique offers high precision, accuracy, and provides automated output. PMID- 19122738 TI - Effects of a series of triorganotins on ATP levels in human natural killer cells. AB - Natural killer (NK) cells are our initial immune defense against viral infections and cancer development. Thus, agents that are able to interfere with their function increase the risk of cancer and/or infection. A series of triorganotins, (trimethyltin (TMT), dimethylphenyltin (DMPT), methyldiphenyltin (MDPT), and triphenyltin (TPT)) have been shown to decrease the lytic function of human NK cells. TPT and MDPT were much more effective than DMPT or TMT at reducing lytic function. This study investigates the role that decreased ATP levels may play in decreases in the lytic function of NK cells induced by these OTs. A 24 h exposure to as high as 10 muM TMT caused no decrease in ATP levels even though this level of TMT caused a greater than 75% loss of lytic function. TPT at 200 nM caused a decrease in ATP levels of about 20% while decreasing lytic function by greater than 85%. There was no association between ATP levels and lytic function for any of the compounds when NK cells were exposed for 1h or 24 h. However, after a 48 h exposure to both DMPT and TPT decreased lytic function was associated with decreased ATP levels. There was an association between decreased lytic function and decreased ATP levels after a 6 day exposure to each of the four compounds. These studies indicate that the loss of lytic function seen after 1 h and 24 h exposures to this series of organotins cannot be accounted for by decreases in ATP. However, after longer exposures loss of lytic function may be in part be attributable to inadequate ATP levels. PMID- 19122737 TI - In vivo fluorescent optical imaging of cytotoxic T lymphocyte migration using IRDye800CW near-infrared dye. AB - We describe a method to measure in vivo migration of human T cells by using the near-infrared (NIR) dye IRDye800CW. Labeling of Epstein-Barr virus-specific T cells with IRDye800CW did not affect viability, proliferation, or T cell function. Following tail vein injection into mice bearing subcutaneous tumors, the NIR signal could be measured in vivo at the tumor site. Analysis of tumors revealed T cell infiltration and an increased NIR signal, confirming T cell migration. To test specific migration with IRDye800CW, tumors were modified to express CCL5 to measure site-specific migration. The NIR signal was increased at CCL5-secreting tumors compared with control tumors. Together, these data suggest that IRDye800CW may be used to study the trafficking of T cells in a small animal model and may have potential as a short-term reporter molecule for human immunotherapy studies. PMID- 19122739 TI - Individual differences in the conceptualization of food across eating contexts. AB - Individual differences in food-related knowledge structures were explored by applying schema theory to examine the categories 42 adults used to classify foods across four eating contexts. Food card-sort labels were organized into 12 categories, category salience for each person was evaluated, and cluster analysis was used to identify clusters of participants according to the salience of their categories. Clusters were further evaluated for complexity and consistency of category use across contexts. Seven food schema clusters were identified. Meal/time and Routine categories were the most salient overall and were used by most clusters. Well-being, Person, Source, Convenience, Meal component, and Food group categories varied in salience across clusters. The complexity and consistency of the food categories participants used across the contexts varied among the clusters. This study provided insight about cognitions that may underlie food-choice behaviors. Understanding individuals' food schemas could help nutrition professionals tailor messages to maximize health impact. PMID- 19122740 TI - A Diffusion Model Account of Criterion Shifts in the Lexical Decision Task. AB - Performance in the lexical decision task is highly dependent on decision criteria. These criteria can be influenced by speed versus accuracy instructions and word/nonword proportions. Experiment 1 showed that error responses speed up relative to correct responses under instructions to respond quickly. Experiment 2 showed that that responses to less probable stimuli are slower and less accurate than responses to more probable stimuli. The data from both experiments support the diffusion model for lexical decision (Ratcliff, Gomez, & McKoon, 2004). At the same time, the data provide evidence against the popular deadline model for lexical decision. The deadline model assumes that "nonword" responses are given only after the "word" response has timed out - consequently, the deadline model cannot account for the data from experimental conditions in which "nonword" responses are systematically faster than "word" responses. PMID- 19122741 TI - Amyloidosis 2008 BMT Tandem Meetings (February 13-17, San Diego). PMID- 19122742 TI - Cancer Stem Cells in Hematopoietic Malignancies. AB - Most hematopoietic malignancies are comprised of cells that are functionally heterogeneous with only a subset being responsible for tumor maintenance. These cancer stem cells are so named because they possess qualities reminiscent of normal tissue stem cells including self-renewal, prolonged survival, and the ability to give rise to cells with more differentiated characteristics. Effort is now focused on identifying cancer stem cells in various hematopoietic malignancies, and defining the cells of origin such that the stepwise accumulation of genetic/epigenetic events necessary for cancer stem cell development can be delineated. A detailed understanding of these processes could lead to development of therapeutics that more effectively treat hematopoietic malignancies and potentially other cancers. PMID- 19122744 TI - Role of oxidative stress in the promoting activities of pcbs. AB - PCBs are organic pollutants that persist and bioaccumulate in the environment. These chemicals induce and promote liver tumors in rodents. Previous studies have shown that they increase oxidative stress in the liver, including lipid peroxidation, oxidative DNA damage, and NF-kappaB activation. The objective of these studies was to determine if the promoting activities of PCBs could be inhibited by dietary antioxidants (vitamin E, selenium, or phytochemicals) or by knocking out the p50 subunit of NF-kappaB. In the antioxidant studies, female rats were first injected with DEN (150 mg/kg) and then administered 4 biweekly i.p. injections (300 MUmol/kg/injection) of PCB-77, PCB-153, or vehicle; the number and volume of placental glutathione S-transferase (PGST)-positive foci were then quantified. Vitamin E did not influence the promoting activities of PCBs. Increasing dietary selenium above the recommended intake increased the number of foci induced but decreased their volume. Most of the phytochemicals examined (N-acetyl cysteine, beta-carotene, resveratrol, EGCG) had no significant effect on the promoting activity of PCB-77. Ellagic acid increased and lycopene decreased the number of foci; ellagic acid, CoQ(10), and curcumin decreased the volume of foci. In the NF-kappaB knockout study, male mice were first injected with DEN (90 mg/kg); controls not receiving DEN were also studied. Both p50 -/- and wild-type mice were then injected biweekly 20 times with PCB-153 (300 (MUmol/kg). In DEN-treated and DEN + PCB-treated mice, the incidence of tumors was lower in the p50 -/- mice than in wild-type mice. In mice receiving PCB-153, the tumor incidence and tumor volume were higher. The volume of tumors that were positive for glutamine synthetase was increased in mice administered PCB-153. This study shows that the promotion of hepatocarcinogenesis by PCBs is largely unaffected by dietary antioxidants but is diminished when NF-kappaB activation is impaired by the absence of the p50 subunit. PMID- 19122745 TI - K-space spatial low-pass filters can increase signal loss artifacts in Echo Planar Imaging. AB - Effective transverse relaxation rate (T(2)*)-weighted echo-planar imaging (EPI) is extensively used for functional magnetic resonance imaging (fMRI), because of its high speed and good sensitivity to the blood oxygenation level-dependent (BOLD) signal. Nevertheless, its use is limited in areas with severe static magnetic field inhomogeneities that cause frequency shifts and T(2)* relaxation related distortions of the MR signal along the time-domain (k-space) trajectory, resulting in disperse time-domain signals and generating susceptibility-induced signal losses. Echo planar images are commonly smoothed with k-space spatial low pass filters to improve the signal-to-noise ratio (SNR) and reduce reconstruction artifacts. Here, we show that when such filters are applied to the dispersed echo signals (not perfectly centered in k-space), part of the image information from the object is removed, thereby enhancing signal-loss artifacts in the images. To avoid this artifact, the dispersed echo signal has to be refocused before k-space filtering. PMID- 19122746 TI - The role of a coda consonant as error trigger in repetition tasks. AB - Speech errors are known to exhibit an intrusion bias in that segments are added rather than deleted; also, a shared final consonant can cause an interaction of the initial consonants. A principled connection between these two phenomena has been drawn in a gestural account of errors: Articulatory measures revealed a preponderance of errors in which both the target and intruding gesture are co produced, instead of one replacing the other. This gestural intrusion bias has been interpreted as an errorful coupling of gestures in a dynamically stable coordination mode (1:1, in-phase), triggered by the presence of a shared coda consonant. Capturing tongue motion with ultrasound, the current paper investigates whether shared gestural composition other than a coda can trigger gestural co-production errors. Subjects repeated two-word phrases with alternating initial stop or fricative consonants in a coda condition (e.g., top cop), a nocoda condition (e.g., taa kaa) and a three-word phrase condition (e.g., taa kaa taa). The no-coda condition showed a lower error rate than the coda condition. The three-word phrase condition elicited an intermediate error rate for the stop consonants, but a high error rate for the fricative alternations. While all conditions exhibited both substitution and co-production errors, a gestural intrusion bias emerged mainly for the coda condition. The findings suggest that the proportion of different error types (substitutions, co production errors) differs as a function of stimulus type: not all alternating stimulus patterns that trigger errors result in an intrusion bias. PMID- 19122747 TI - Stability of Temporal Contrasts across Speaking Styles in English and Croatian. AB - In this paper we investigate the effect of clear speech, a distinct, listener oriented, intelligibility-enhancing mode of speech production, on vowel and stop consonant contrasts along the temporal dimension in English and Croatian. Our previous work has shown that, in addition to enhancing the overall acoustic salience of the speech signal through a decrease in speaking rate and expansion of pitch range, clear speech modifications increased the spectral distances between vowel categories in both languages despite the different sizes of their vowel inventories (+10 in English, 5 in Croatian). Here, we examine how clear speech affects the duration of English tense ('long') vs. lax ('short') vowels, English vowels preceding voiced ('long') vs. voiceless ('short') coda stops, Croatian long vs. short vowels and Croatian and English VOT duration for voiced and voiceless stops. Overall, the results showed that the proportional distance between the 'short' and 'long' vowel categories and between the voiced and voiceless stop categories was remarkably stable across the two speaking styles in both languages. These results suggest that, in combination with the spectral enhancement of vowel contrasts, language-specific pronunciation norms along the temporal dimension are maintained in clear and conversational speech. PMID- 19122748 TI - The Effects of Unequal Reward Distributions on Cooperative Problem Solving by Cottontop Tamarins (Saguinus oedipus). AB - Cooperation among non-human animals has been the topic of much theoretical and empirical research, but few studies have examined systematically the effects of various reward payoffs on cooperative behaviour. Here, we presented heterosexual pairs of cooperatively breeding cottontop tamarins with a cooperative problem solving task. In a series of four experiments, we examined how the tamarins' cooperative performance changed under conditions in which (a) both actors were mutually rewarded, (b) both actors were rewarded reciprocally across days, (c) both actors competed for a monopolizable reward and (d) one actor repeatedly delivered a single reward to the other actor. The tamarins demonstrated sensitivity to the reward structure, exhibiting the greatest percentage of trials solved and shortest latency to solve the task in the mutual reward experiment and the lowest percentage of trials solved and longest latency to solve the task in the experiment in which one actor was repeatedly rewarded. However, even in the experiment in which the fewest trials were solved, the tamarins still solved 46 +/- 12% of trials and little to no aggression was observed among partners following inequitable reward distributions. The tamarins did, however, exhibit selfish motivation in each of the experiments. Nevertheless, in all experiments, unrewarded individuals continued to cooperate and procure rewards for their social partners. PMID- 19122749 TI - Maintaining Sibling Relationships for Children in Foster and Adoptive Placements. AB - To better understand the nature of sibling relationships among children in foster or adoptive placement and the challenges and processes involved in maintaining these relationships, we conducted an exploratory analysis of data collected from semi-structured interviews with caregivers of 14 foster and adopted children in San Diego County. We identified three patterns of placement histories and living situations which appeared to impact the degree of contact maintained with siblings: (1) children who had never lived together and were not currently placed together; (2) children who at some point lived with or were placed together with their siblings, but were now separated from them; and (3) children who had lived with their siblings all their life and were placed together with at least some of their siblings at the time of the interview. Children's current living situations and placement histories, caregivers' experiences and perceptions of feasibility and desirability of sibling contact, and the sibling relationship itself are primary determinants in the development and maintenance of contact between siblings. Implications for child welfare policy and practice are discussed. PMID- 19122750 TI - Vitamin B(12) and alpha-Ribonucleosides. PMID- 19122751 TI - Preschoolers' oculomotor behaviour during their observation of an action task. AB - Adult and child oculomotor behaviour was measured during the observation of a naturalistic action task. Adult and 4-year-old participants viewed a video presentation of an actor making a root-beer float. Eye movements were monitored to examine fixation patterns during a series of means-end action sequences. Look ahead fixations (i.e., gaze anticipation) were measured to determine if children would saccade to the goal site prior to the completion of each action sequence as adults do. Look-ahead fixations were observed in both the adult and child groups, with no significant age differences found in levels of gaze anticipation. PMID- 19122752 TI - Objective Ratings of Relationship Skills across Multiple Domains as Predictors of Marital Satisfaction Trajectories. AB - Expanding upon social-learning and vulnerability-stress-adaptation approaches to marriage, the impact of multiple dyadic behaviors on marital satisfaction trajectories was examined in 101 couples. Semi-structured interviews were administered separately to husbands and wives at 3 months of marriage. Interviewers generated objective ratings for five domains: emotional closeness/intimacy, sexual intimacy/sensuality, interspousal support, decision making/relational control, and communication/conflict management. Marital satisfaction was assessed four times over three years. Dyadic behaviors were associated with initial levels and rates of change in satisfaction, demonstrating the unique contributions of each relational skill on marital development. For husbands, sexual intimacy was the strongest predictor of change whereas for wives, communication/conflict management was the strongest predictor of change compared to other domains. Theoretical, methodological and clinical implications are discussed. PMID- 19122753 TI - Influence of viscosity on the reflection and transmission of an acoustic wave by a periodic array of screens. The general 3-D problem. AB - An analysis is presented of the diffraction of a pressure wave by a periodic grating including the influence of the air viscosity. The direction of the incoming pressure wave is arbitrary. As opposed to the classical nonviscous case, the problem cannot be reduced to a plane problem having a definite 3-D character. The system of partial differential equations used for solving the problem consists of the compressible Navier-Stokes equations associated with no-slip boundary conditions on solid surfaces. The problem is reduced to a system of two hypersingular integral equations for determining the velocity components in the slits' plane and a hypersingular integral equation for the normal component of velocity. These equations are solved by using Galerkin's method with some special trial functions. The results can be applied in designing protective screens for miniature microphones realized in MEMS technology. In this case, the physical dimensions of the device are on the order of the viscous boundary layer so that the viscosity cannot be neglected. The analysis indicates that the openings in the screen should be on the order of 10 microns in order to avoid excessive attenuation of the signal. This paper also provides the variation of the transmission coefficient with frequency in the acoustical domain. PMID- 19122754 TI - The Introduction of Bioptic Driving in The Netherlands. AB - BACKGROUND: In many states of the U.S.A., people with moderately reduced visual acuity e.g., 20/50 - 20/200) can legally drive with the aid of a small, spectacle mounted ("bioptic") telescope. We conducted a demonstration project to assess the viability of implementing bioptic driving in The Netherlands. In this paper we describe the framework of the project from conception through to realization of our primary objective - the introduction of bioptic driving as a legal option for visually impaired people in The Netherlands. METHODS: The project was based on bioptic driving programs in the U.S.A., which were adapted to fit within current driving training and assessment practices in The Netherlands. The project convened a consortium of organizations including the Netherlands Bureau of Driving Skills Certificates (CBR), service organizations for the visually impaired, and research departments at universities investigating driving and vision. All organizations were educated about bioptic driving and participating professionals were trained in their specific aspects of the project. Media publicity led to significant interest and helped recruitment that enabled the screening and selection of potential participants. OUTCOMES: The project demonstrated that people with moderately reduced visual acuity can be trained to achieve an adequate level of proficient and safe driving (as assessed by the local official driving licensing professionals) when using a bioptic telescope for the road conditions in the Netherlands. Based on the successful project outcomes, a request was made to the Minister to allow bioptic driving in the Netherlands. This request has been accepted; the legal procedures for implementation are in process. PMID- 19122755 TI - Synthesis of a Homologous Series of Side Chain Extended Orthogonally-Protected Aminooxy-Containing Amino Acids. AB - Practical methodology is reported for the synthesis of a homologous series of side chain extended amino acids containing aminooxy functionality bearing orthogonal protection suitable for Fmoc peptide synthesis. These reagents may be useful for the preparation of libraries containing fragments joined by peptide linkers. PMID- 19122756 TI - Beyond Self-Report in the Study of Hedonic and Eudaimonic Well-Being: Correlations with Acquaintance Reports, Clinician Judgments and Directly Observed Social Behavior. PMID- 19122757 TI - A Cognitive Cascade in Infancy: Pathways from Prematurity to Later Mental Development. AB - Using data from a longitudinal study of preterms and full-terms, the present study examined the structure of infant cognition at 12 months, the extent to which five 12-month abilities (attention, speed, recognition, recall, and representational competence) mediated the relation from prematurity to mental development at 2 - 3 years, and how continuity and change in infant information processing from 7 to 12 months affected later outcome. The results indicated that 12-month measures of infant information processing completely mediated the effect of prematurity on outcome and the infant measures form a 'cognitive cascade,' similar to that seen at 7 months, in which the two more elementary abilities (attention and speed) influenced the more complex ones, which in turn influenced later cognition. Additionally, despite cross-age stability, 7- month assessments contribute to outcome independently of their 12-month counterparts, suggesting that infant abilities undergo important developmental transformations in the second half of the first year of life. PMID- 19122758 TI - Stereoselective synthesis of imidazolidin-2-ones via Pd-catalyzed alkene carboamination. Scope and limitations. AB - A method for the synthesis of imidazolidin-2-ones from N-allylureas and aryl or alkenyl bromides via Pd-catalyzed carboamination reactions is described. The N allylurea precursors are prepared in one step from readily available allylic amines and isocyanates, and the Pd-catalyzed reactions effect the formation of a C-C bond, a C-N bond, and up to two stereocenters in a single step. Good diastereoselectivities are obtained for the conversion of substrates bearing allylic substituents to 4,5-disubstituted imidazolidin-2-ones, and excellent selectivity for the generation of products resulting from syn-addition across the alkene is observed when substrates derived from cyclic alkenes or E-1,2 disubstituted alkenes are employed. A brief discussion of reaction mechanism and product stereochemistry is presented. PMID- 19122759 TI - Older Youth Leaving the Foster Care System: Who, What, When, Where, and Why? PMID- 19122760 TI - Insomnia: Prevalence, Impact, Pathogenesis, Differential Diagnosis, and Evaluation. PMID- 19122761 TI - Assessment of Density Functionals, Semiempirical Methods, and SCC-DFTB for Protonated Creatinine Geometries. AB - Creatinine concentrations in blood and urine can be used to detect renal insufficiencies and muscle diseases, but current chemical sensors cannot measure creatinine with sufficient selectivity and robustness because they lack a receptor that binds protonated creatinine (creatininium) selectively enough. As a first step toward identifying potential receptors for creatininium, we examine the accuracy of density functional theory (DFT) and wave function theory (WFT) calculations for creatininium cation geometries, evaluated against reference parameters from experiment. We tested twenty-one local and nonlocal density functionals, Hartree-Fock theory, four semiempirical molecular orbital (SEMO) methods of the neglect of differential overlap (NDO) type, and one self consistent-field nonorthogonal tight-binding method (SCC-DFTB) as implemented in two closely related software packages. DFT and HF calculations were carried out with the MG3S augmented polarized triple-zeta basis set. We find that DFT significantly outperforms SEMO methods for our dataset, and both SCC-DFTB releases we tested (which gave almost identical results) were less accurate than 81% of the density functionals evaluated. The top five functionals for the creatininium structures we examined were MPW1B95, PBEh, mPW1PW, SVWN5, and B97-2, with MMUEs in bond length of 0.0126 A, 0.0129 A, 0.0133 A, 0.0142 A, and 0.0144 A respectively, which indicates that all five functionals are similarly suitable for creatininium. The popular B3LYP functional has a MMUE of 0.0178 A, which ranks it 16(th) overall. B3LYP also performs less favorably than the best local functional (SVWN5), which is less expensive. PMID- 19122762 TI - Synthesis of Fluorescent Probes based on Stilbenes and Diphenylacetylenes targeting beta-Amyloid Plaques. AB - Three fluorescent probes were synthesized aiming for optical imaging to detect amyloid plaques present in patients with Alzheimer's disease (AD). These compounds were prepared via Sonogashira coupling of a well-defined fluorophore (4 bora-3a, 4a-diaza s-indacene, BODIPY) with the pharmacophore possessing either a stilbene or a diphenylacetylene moiety. Different polyethylene glycol chain lengths were used as linkers between the fluorophore and the pharmacophore to adjust the lipophilicity of these probes. These compounds exhibit strong fluorescence emission between 665-680 nm and have very high extinction coefficients comparable to the parent fluorophore, BODIPY dye. PMID- 19122763 TI - Comparing Outcomes for Youth in Treatment Foster Care and Family-style Group Care. AB - Group care programs are often criticized for producing poor outcomes, especially in light of community-based alternatives like treatment foster care that have a stronger evidence base. In this study, data from Girls and Boys Town were used to compare outcomes of youth in treatment foster care (n=112) and group care (n=716) using propensity score matching, a method that can minimize selection bias in nonrandomized designs. Eighteen background covariates were used to develop propensity scores for the likelihood of receiving treatment foster care rather than group care. Several matching methods generated balanced samples on which the outcomes were compared. Results found that group care youth were more likely to be favorably discharged, more likely to return home, and less likely to experience subsequent placement in the first six months after discharge. Legal involvement and residing in a home-like environment at follow-up did not differ. Positive outcomes for group care youth suggest that family-style group care programs may promote effectiveness. PMID- 19122764 TI - Coupling of Vinyl Aziridines and Isocyanates. AB - Thermal coupling of vinyl aziridines and phenyl isocyanate was evaluated. Although oxazolidinone products were predominant, some reactions afforded a seven membered ring heterocycle. When Ni/IMes was employed as a catalyst, a wider array of vinyl aziridines underwent coupling reactions. The Ni catalyzed reactions generally afforded vinyl imidazolidinones as major products. PMID- 19122766 TI - BETTER STATISTICS FOR BETTER DECISIONS: REJECTING NULL HYPOTHESES STATISTICAL TESTS IN FAVOR OF REPLICATION STATISTICS. AB - Despite being under challenge for the past 50 years, null hypothesis significance testing (NHST) remains dominant in the scientific field for want of viable alternatives. NHST, along with its significance level p, is inadequate for most of the uses to which it is put, a flaw that is of particular interest to educational practitioners who too often must use it to sanctify their research. In this article, we review the failure of NHST and propose p(rep), the probability of replicating an effect, as a more useful statistic for evaluating research and aiding practical decision making. PMID- 19122765 TI - Pediatric Cardiomyopathy as a Chronic Disease: A Perspective on Comprehensive Care Programs. AB - Substantial numbers of children with cardiomyopathy are now surviving into adulthood, making it essentially a chronic disease. As a chronic condition, it may be best treated through comprehensive, multidisciplinary treatment programs. Such programs have improved health outcomes and reduced costs in managing other pediatric chronic diseases and heart failure in adults, but the treatment and cost implications of programs for managing pediatric cardiomyopathy are unknown. We investigated the treatment and cost implications of establishing such programs by reviewing cost-effectiveness studies of similar programs, estimating the current inpatient costs of this diagnosis, and interviewing experts in the field about the need and desirability of these programs. According to our findings, comprehensive pediatric heart failure programs do exist, but they have not been evaluated or even described in the literature. Consensus among experts in the field is that such programs are highly desirable, and similar programs have reported tremendous cost savings through early and intensive management: the return on investment has been as high as 22 to 1. Another study reported that mean length of stay decreased from 83.9 to 10.6 days, mean annual admissions decreased from 2,796 to 1,622, and median hospital charges decreased from $26.1 million to $14.6 million. In conclusion, limited experience and strong circumstantial evidence suggest that, despite substantial costs, comprehensive multidisciplinary pediatric heart failure programs would be highly cost-effective and beneficial to patients, families, and institutions alike. PMID- 19122767 TI - NATURALIZATION OF U.S. IMMIGRANTS: HIGHLIGHTS FROM TEN COUNTRIES. AB - The saga of U.S. immigrant naturalization is merely sketched for about 25 million immigrants entered in three decades of renewed immigration. This study documents naturalization outcomes for immigrants from ten major countries of origin, using administrative records on immigrants and naturalizations. Following the 1978-1987 admission cohorts for the first decade or more of permanent residence, this study finds significant covariate effects on the timing of naturalization by origin, mode of entry, and immigrant visa class, net other influences of demographic and background characteristics. Immigrants from the Philippines, Vietnam, and China, naturalized more quickly than immigrants from India, Korea, Cuba, Colombia, Jamaica, the Dominican Republic, and Mexico. Those who adjusted from statuses as nonimmigrants, refugees, or asylees became naturalized citizens more quickly. Those immigrants with employment-sponsorship naturalized faster than family sponsored immigrants. Spouses of citizens, spouses of permanent residents, spouses of siblings of citizens, and spouses of sons and daughters of citizens naturalized faster than some other immigrants. Gender was not significant in the multivariate analysis but further research will more fully explore sex-specific variation in the timing of naturalization given likely variation in women's representation by origin and admission categories. PMID- 19122768 TI - How to measure quality of life in integrative oncology research. AB - Quality of life is an important focus of integrative oncology research. Choice of an appropriate way to measure quality of life is therefore a key concern. A common error of researchers has been to focus on whether a quality of life measure has been "validated". This may be a necessary condition for choice of endpoint, but it is certainly not sufficient. It is easily shown that interventions that undoubtedly improve quality of life, such as Epogen for chemotherapy anemia, will not have an important impact on items in some validated quality of life scales (e.g. "I trust my doctor") and therefore will not improve quality of life scores. Investigators are advised to move beyond "validation" as a criterion for choosing quality of life endpoints and focus instead on the specific content of different potential measures. A particularly important concern is whether a prior trial showing a statistically significant difference on a specific quality of life scale is published, as this would demonstrate that scores on the scale will improve if patients respond to treatment. In many cases, it is preferable to use specific symptoms, such as pain or fatigue, as opposed to more generic quality of life scales. PMID- 19122769 TI - CONTRIBUTIONS OF THE MOTHER-INFANT RELATIONSHIP TO DISSOCIATIVE, BORDERLINE, AND CONDUCT SYMPTOMS IN YOUNG ADULTHOOD. AB - Recent high-risk longitudinal studies have documented a unique contribution of the quality of the early mother-child relationship to diverse forms of psychopathology in young adulthood, even with family economic status, later traumatic experiences, and some genetic factors controlled. In addition, measures of attachment-related deviations in caregiver-infant interaction predict more than measures of infant attachment behavior alone. This article reviews those findings in the context of cross-disciplinary thinking on the importance of shared subjectivities in human evolution and development and in the context of recent studies beginning to map the intersection between processes of interaction and the development of the child's propensities to share mental states with others. PMID- 19122770 TI - CHRONOBIOLOGY OF HIGH BLOOD PRESSURE. AB - BIOCOS, the project aimed at studying BIOlogical systems in their COSmos, has obtained a great deal of expertise in the fields of blood pressure (BP) and heart rate (HR) monitoring and of marker rhythmometry for the purposes of screening, diagnosis, treatment, and prognosis. Prolonging the monitoring reduces the uncertainty in the estimation of circadian parameters; the current recommendation of BIOCOS requires monitoring for at least 7 days. The BIOCOS approach consists of a parametric and a non-parametric analysis of the data, in which the results from the individual subject are being compared with gender- and age-specified reference values in health.Chronobiological designs can offer important new information regarding the optimization of treatment by timing its administration as a function of circadian and other rhythms.New technological developments are needed to close the loop between the monitoring of blood pressure and the administration of antihypertensive drugs. PMID- 19122772 TI - In vivo imaging of transport and biocompatibility of single silver nanoparticles in early development of zebrafish embryos. AB - Real-time study of the transport and biocompatibility of nanomaterials in early embryonic development at single-nanoparticle resolution can offer new knowledge about the delivery and effects of nanomaterials in vivo and provide new insights into molecular transport mechanisms in developing embryos. In this study, we directly characterized the transport of single silver nanoparticles into an in vivo model system (zebrafish embryos) and investigated their effects on early embryonic development at single-nanoparticle resolution in real time. We designed highly purified and stable (not aggregated and no photodecomposition) nanoparticles and developed single-nanoparticle optics and in vivo assays to enable the study. We found that single Ag nanoparticles (5-46 nm) are transported into and out of embryos through chorion pore canals (CPCs) and exhibit Brownian diffusion (not active transport), with the diffusion coefficient inside the chorionic space (3 x 10(-9) cm(2)/s) approximately 26 times lower than that in egg water (7.7 x 10(-8) cm(2)/s). In contrast, nanoparticles were trapped inside CPCs and the inner mass of the embryos, showing restricted diffusion. Individual Ag nanoparticles were observed inside embryos at each developmental stage and in normally developed, deformed, and dead zebrafish, showing that the biocompatibility and toxicity of Ag nanoparticles and types of abnormalities observed in zebrafish are highly dependent on the dose of Ag nanoparticles, with a critical concentration of 0.19 nM. Rates of passive diffusion and accumulation of nanoparticles in embryos are likely responsible for the dose-dependent abnormalities. Unlike other chemicals, single nanoparticles can be directly imaged inside developing embryos at nanometer spatial resolution, offering new opportunities to unravel the related pathways that lead to the abnormalities. PMID- 19122773 TI - A HAUSDORFF-BASED NOE ASSIGNMENT ALGORITHM USING PROTEIN BACKBONE DETERMINED FROM RESIDUAL DIPOLAR COUPLINGS AND ROTAMER PATTERNS. AB - High-throughput structure determination based on solution Nuclear Magnetic Resonance (NMR) spectroscopy plays an important role in structural genomics. One of the main bottlenecks in NMR structure determination is the interpretation of NMR data to obtain a sufficient number of accurate distance restraints by assigning nuclear Overhauser effect (NOE) spectral peaks to pairs of protons. The difficulty in automated NOE assignment mainly lies in the ambiguities arising both from the resonance degeneracy of chemical shifts and from the uncertainty due to experimental errors in NOE peak positions. In this paper we present a novel NOE assignment algorithm, called HAusdorff-based NOE Assignment (HANA), that starts with a high-resolution protein backbone computed using only two residual dipolar couplings (RDCs) per residue37, 39, employs a Hausdorff-based pattern matching technique to deduce similarity between experimental and back computed NOE spectra for each rotamer from a statistically diverse library, and drives the selection of optimal position-specific rotamers for filtering ambiguous NOE assignments. Our algorithm runs in time O(tn(3) +tn log t), where t is the maximum number of rotamers per residue and n is the size of the protein. Application of our algorithm on biological NMR data for three proteins, namely, human ubiquitin, the zinc finger domain of the human DNA Y-polymerase Eta (pol eta) and the human Set2-Rpb1 interacting domain (hSRI) demonstrates that our algorithm overcomes spectral noise to achieve more than 90% assignment accuracy. Additionally, the final structures calculated using our automated NOE assignments have backbone RMSD < 1.7 A and all-heavy-atom RMSD < 2.5 A from reference structures that were determined either by X-ray crystallography or traditional NMR approaches. These results show that our NOE assignment algorithm can be successfully applied to protein NMR spectra to obtain high-quality structures. PMID- 19122774 TI - A Novel Method of Fabricating Convoluted Shaped Electrode Arrays for Neural and Retinal Prostheses. AB - A novel fabrication technique has been developed for creating high density (6.25 electrodes/mm(2)), out of plane, high aspect ratio silicon-based convoluted microelectrode arrays for neural and retinal prostheses. The convoluted shape of the surface defined by the tips of the electrodes could compliment the curved surfaces of peripheral nerves and the cortex, and in the case of retina, its spherical geometry. The geometry of these electrode arrays has the potential to facilitate implantation in the nerve fascicles and to physically stabilize it against displacement after insertion. This report presents a unique combination of variable depth dicing and wet isotropic etching for the fabrication of a variety of convoluted neural array geometries. Also, a method of deinsulating the electrode tips using photoresist as a mask and the limitations of this technique on uniformity are discussed. PMID- 19122776 TI - A Theory-Based Model for Predicting Adherence to Guidelines for Screening Mammography among Women Age 40 and Older. AB - OBJECTIVE: To examine theory-based selected factors associated with adherence to mammography screening guidelines in a surveillance database. METHODS: Data from Colorado Mammography Project (CMAP) from 1994-1998 was extracted and analyzed by using SAS statistical software. Based on the Health Belief Model and Behavioral Model of Health Services Utilization a prediction model was developed to examine the mammography utilization patterns and factors influencing the adherence to screening guidelines. RESULTS: Out of 27,778 women, 41.4% were adherent with mammography screening guidelines. According to the model tested in this study, race/ethnicity (Black vs White, OR=0.76, 95% CI=0.64-0.91); educational attainment (high school vs < high school, OR= 1.10, 95% CI= 1.04-1.18), college graduate vs < high school (OR=1.33, 95% CI=1.25-1.42); insurance status, (any coverage vs no coverage (OR=1.62, 95% CI=1.25-2.12); and community economic status as defined by median income by zip code of residence ($15,000-$24,999 vs <$15,000, OR=0.84, 95% CI=0.76-0.94; >$55,000 vs <$15,000, OR 1.14, 95% CI=1.03 1.26) were statistically significant predictors of adherence to guidelines. Interaction between age and family history of breast cancer was statically significant. Younger females with a family history of breast cancer were less likely to be adherent than their counterparts without a family history (OR=0.93, 95% CI=0.90-0.96). Inclusion or exclusion of women aged 70 years and older did not change the outcome of the analysis. CONCLUSION: The prediction model variables such as race/ethnicity, age and family history of breast cancer, educational level and community economic status, are associated with adherence status. Family history of breast cancer needs to be examined very carefully in future studies as it may play negative role in adherence to screening mammography. PMID- 19122777 TI - Capillary-contacting horizontal cells in the rodent retina. AB - Horizontal cells, the interneurons in the distal retina, provide feedback control of the photoreceptor synaptic output at the first synapse in the visual pathway. This article, using immunohistological and confocal microscopy techniques, presents anatomical evidence that in rat and mouse retina, the horizontal cell processes are in contact with retinal capillaries as are retinal glial cells. This glia-like property of horizontal cells in these two species, also previously reported in tree shrews by Knabe and Ochs (1999), appear to be a more common theme for these neurons than previously appreciated. PMID- 19122778 TI - Persistent transport barrier on the West Florida Shelf. AB - Analysis of drifter trajectories in the Gulf of Mexico has revealed the existence of a region on the southern portion of the West Florida Shelf (WFS) that is not visited by drifters that are released outside of the region. This so-called "forbidden zone" (FZ) suggests the existence of a persistent cross-shelf transport barrier on the southern portion of the WFS. In this letter a year-long record of surface currents produced by a Hybrid-Coordinate Ocean Model simulation of the WFS is used to compute Lagrangian coherent structures (LCSs), which reveal the presence of a persistent cross-shelf transport barrier in approximately the same location as the boundary of the FZ. The location of the cross-shelf transport barrier undergoes a seasonal oscillation, being closer to the coast in the summer than in the winter. A month-long record of surface currents inferred from high-frequency (HF) radar measurements in a roughly 60 km * 80 km region on the WFS off Tampa Bay is also used to compute LCSs, and these also reveal the presence of transient transport barriers. While the HF-radar-derived transport barriers cannot be unambiguously linked to the boundary of the FZ, this analysis does demonstrate the feasibility of monitoring transport barriers on the WFS using a HF-radar-based measurement system. The implications of a persistent cross shelf transport barrier on the WFS for the development of harmful algal blooms on the shoreward side of the barrier are considered. PMID- 19122779 TI - Soluble Guanylyl Cyclases in Invertebrates: Targets for NO and O(2). PMID- 19122780 TI - Decisional conflict among patients who accept or decline participation in phase I oncology studies. AB - WE COMPARED DECISIONAL CONFLICT among adults with advanced cancer who had accepted or declined participation in phase I cancer clinical trials. Respondents completed a 121-item questionnaire that included the Decisional Conflict Scale (DCS), which was designed to measure uncertainty in making health decisions. We used standardized effect sizes to compare the DCS scores of accepters (n = 250) and decliners (n = 65). Accepters had lower decisional conflict than decliners overall (d = 0.42; 95% confidence interval, 0.17--0.68) and on all subscales. Whether greater decisional conflict among decliners represents suboptimal decision-making and is reason for bioethical concern depends on how the results are interpreted. We offer three scenarios to explain the differences and describe opportunities for future empirical work. PMID- 19122782 TI - Transcranial magnetic stimulation in Clinical Pharmacology. AB - Transcranial magnetic stimulation (TMS) is a safe and easy technique for stimulating neurons in the human central nervous system. Studies combining TMS with drugs in healthy subjects and patients have advanced our knowledge of how TMS activates brain circuits and led to new techniques for evaluating the function of specific systems. For example, TMS techniques can detect effects on axon membranes, glutamatergic and GABAergic synapses and the influence of catecholaminergic systems, as well as group differences due to genetic variations in the response to drugs. With this knowledge base, TMS can now be used to explore and compare the effects of drugs on brain systems and may also serve as a surrogate for behavioral responses in clinical trials. PMID- 19122781 TI - Wnt signaling in eye organogenesis. AB - The vertebrate eye consists of multiple tissues with distinct embryonic origins. To ensure formation of the eye as a functional organ, development of ocular tissues must be precisely coordinated. Besides intrinsic regulators, several extracellular pathways have been shown to participate in controlling critical steps during eye development. Many components of Wnt/Frizzled signaling pathways are expressed in developing ocular tissues, and substantial progress has been made in the past few years in understanding their function during vertebrate eye development. Here, I summarize recent work using functional experiments to elucidate the roles of Wnt/Frizzled pathways during development of ocular tissues in different vertebrates. PMID- 19122783 TI - Epidemiologic and behavioral characterization of knowledge of condom use and modeling among military personnel. AB - How accurately condoms are being used vary across populations and knowledge of the factors determining its proper use remains unclear. Knowledge of such differentials and determinants would aid in evaluating the contributions of condom use to HIV epidemic reduction. Baseline data from the Situationally Focused Individual HIV/AIDS intervention to promote HIV protective behavior among 2,213 Nigerian Military Personnel were analyzed. Educational status as a predictor variable was assessed using univariable and multivariable logistic regression model. Compared to those with less than high school education, those with high school and some college education were two times more likely to demonstrate knowledge of condom use and modeling, prevalence odds ratio (POR), 2.32, 95% Confidence Interval (CI) = 1.60-3.37. After adjustment for the relevant covariates, higher education attainment was associated with nonsignificant 62% increase in knowledge and modeling, POR, 1.62, 95% CI = 0.78-3.38. This study is indicative of low knowledge of condom use and modeling among the Nigerian military personnel; as well as a direct correlation between education attainment and knowledge of condom use and modeling. PMID- 19122784 TI - Synthesis of 19-Trideuterated ent-Testosterone and the GABA(A) Receptor Potentiators ent-Androsterone and ent-Etiocholanolone. AB - 19-Trideuteromethyl enantiomers of androgens namely ent-testosterone, ent androsterone and ent-etiocholanolone were prepared by total synthesis. The isotope labeling at the carbon-19 angular methyl group was achieved by using deuterated methyl iodide (99.5% d(3)) for introduction of C-19 before closure of the steroid A-ring. This method yields 19,19,19-trideuterated steroids without increasing the number of steps involved in the total synthesis of ent-androgens. Analysis by mass spectrometry showed no loss of deuterium during incorporation of C-19 into ent-testosterone. The availability of the compounds will enable these ent-androgens to be distinguished by mass spectrometry from their natural enantiomers in future pharmacokinetic and metabolic studies. PMID- 19122785 TI - AN EFFICIENT SYNTHESIS OF TRICYCLIC COMPOUNDS, (+/-)-(4abeta8abeta10aalpha) 1,2,3,4,4a,6,7,8,8a,9,10,10a-DODECAHYDRO-1,1,4a-TRIMETHYL-2-OXOPHENANTHRENE-8a CARBOXYLIC ACID, ITS METHYL ESTER, AND (+/-)-(4abeta,8abeta10aalpha) 3,4,4a,6,7,8,8a,9,10,10a-DECAHYDRO-8a-HYDROXYMETHYL-1,1,4a-TRIMETHYLPHENANTHREN 2(1H)-ONE. PMID- 19122788 TI - CULTURAL DISPLAY RULES DRIVE EYE GAZE DURING THINKING. AB - The authors measured the eye gaze displays of Canadian, Trinidadian, and Japanese participants as they answered questions for which they either knew, or had to derive, the answers. When they knew the answers, Trinidadians maintained the most eye contact, whereas Japanese maintained the least. When thinking about the answers to questions, Canadians and Trinidadians looked up, whereas Japanese looked down. Thus, for humans, gaze displays while thinking are at least in part culturally determined. PMID- 19122789 TI - Change and Stability in the Characteristics of Homicide Victims, Offenders and Incidents During Rapid Social Change. AB - The Russian homicide rate doubled during the 1990s and is now among the highest in the world. During this same period, Russian citizens experienced swift, widespread, and meaningful political, economic, and social change. It is likely that this profound transition altered structural conditions, cultural norms, and interpersonal relations in a way that led to changes in the nature of interpersonal violence. Taking advantage of a unique set of homicide narratives drawn from court and police records in the Udmurt Republic, this study examined stability and change in the distribution of Russian homicide victim, offender, and incident characteristics before and after the fall of the Soviet Union. Odds ratios obtained from logistic regression showed no change in victim characteristics, but substantial changes in several offender and incident characteristics. We discuss the potential mechanisms through which the structural and cultural shifts are resulting in these changes and conclude that the ongoing transition is largely responsible for the changing nature of homicide in Russia. In doing so, we introduce the new term "criminological transition" and suggest that Russia (and perhaps other nations) may have experienced a change in its crime profile in much the same way as we discuss a "demographic transition" in terms of fertility and mortality profiles. PMID- 19122786 TI - Smoking, Genetics and Schizophrenia: Evidence for Self Medication. AB - Schizophrenia is a common mental illness with a high prevalence of smoking. More than 80% of schizophrenics smoke compared to 25% of the general population. Both schizophrenia and tobacco use have strong genetic components, which may overlap. It has been suggested that smoking in schizophrenia may be a form of self medication in an attempt to treat an underlying biological pathology. Smoking normalizes auditory evoked potential and eye tracking deficits in schizophrenia, as well as improving cognitive function. Nicotine acts through a family of nicotinic receptors with either high or low affinity for nicotine. The loci for several of these receptors have been genetically linked to both smoking and to schizophrenia. Smoking changes gene expression for more than 200 genes in human hippocampus, and differentially normalizes aberrant gene expression in schizophrenia. The alpha7* nicotinic receptor, linked to schizophrenia and smoking, has been implicated in sensory processing deficits and is important for cognition and protection from neurotoxicity. Nicotine, however, has multiple health risks and desensitizes the receptor. A Phase I trial of DMXB-A, an alpha7* agonist, shows improvement in both P50 gating and in cognition, suggesting that further development of nicotinic cholinergic drugs is a promising direction in schizophrenia research. PMID- 19122790 TI - Men's Aggression Toward Women: A 10-Year Panel Study. AB - The present study examined the longitudinal course of men's physical and psychological aggression toward a partner across 10 years, using a community sample of young couples (N = 194) from at-risk backgrounds. Findings indicated that men's aggression decreased over time and that women's antisocial behavior and depressive symptoms predicted changes in men's aggression. This suggests the importance of studying social processes within the dyad to have a better understanding of men's aggression toward a partner. PMID- 19122791 TI - Sensitivity Analyses Comparing Time-to-Event Outcomes Existing Only in a Subset Selected Postrandomization. AB - In some randomized studies, researchers are interested in determining the effect of treatment assignment on outcomes that may exist only in a subset chosen after randomization. For example, in preventative human immunodeficiency virus (HIV) vaccine efficacy trials, it is of interest to determine whether randomization to vaccine affects postinfection outcomes that may be right-censored. Such outcomes in these trials include time from infection diagnosis to initiation of antiretroviral therapy and time from infection diagnosis to acquired immune deficiency syndrome. Here we present sensitivity analysis methods for making causal comparisons on these postinfection outcomes. We focus on estimating the survival causal effect, defined as the difference between probabilities of not yet experiencing the event in the vaccine and placebo arms, conditional on being infected regardless of treatment assignment. This group is referred to as the always-infected principal stratum. Our key assumption is monotonicity-that subjects randomized to the vaccine arm who become infected would have been infected had they been randomized to placebo. We propose nonparametric, semiparametric, and parametric methods for estimating the survival causal effect. We apply these methods to the first Phase III preventative HIV vaccine trial, VaxGen's trial of AIDSVAX B/B. PMID- 19122792 TI - Statistical learning of origin-specific statically optimal individualized treatment rules. AB - Consider a longitudinal observational or controlled study in which one collects chronological data over time on a random sample of subjects. The time-dependent process one observes on each subject contains time-dependent covariates, time dependent treatment actions, and an outcome process or single final outcome of interest. A statically optimal individualized treatment rule (as introduced in van der Laan et. al. (2005), Petersen et. al. (2007)) is a treatment rule which at any point in time conditions on a user-supplied subset of the past, computes the future static treatment regimen that maximizes a (conditional) mean future outcome of interest, and applies the first treatment action of the latter regimen. In particular, Petersen et. al. (2007) clarified that, in order to be statically optimal, an individualized treatment rule should not depend on the observed treatment mechanism. Petersen et. al. (2007) further developed estimators of statically optimal individualized treatment rules based on a past capturing all confounding of past treatment history on outcome. In practice, however, one typically wishes to find individualized treatment rules responding to a user-supplied subset of the complete observed history, which may not be sufficient to capture all confounding. The current article provides an important advance on Petersen et. al. (2007) by developing locally efficient double robust estimators of statically optimal individualized treatment rules responding to such a user-supplied subset of the past. However, failure to capture all confounding comes at a price; the static optimality of the resulting rules becomes origin-specific. We explain origin-specific static optimality, and discuss the practical importance of the proposed methodology. We further present the results of a data analysis in which we estimate a statically optimal rule for switching antiretroviral therapy among patients infected with resistant HIV virus. PMID- 19122793 TI - Causal effect models for realistic individualized treatment and intention to treat rules. AB - Marginal structural models (MSM) are an important class of models in causal inference. Given a longitudinal data structure observed on a sample of n independent and identically distributed experimental units, MSM model the counterfactual outcome distribution corresponding with a static treatment intervention, conditional on user-supplied baseline covariates. Identification of a static treatment regimen-specific outcome distribution based on observational data requires, beyond the standard sequential randomization assumption, the assumption that each experimental unit has positive probability of following the static treatment regimen. The latter assumption is called the experimental treatment assignment (ETA) assumption, and is parameter-specific. In many studies the ETA is violated because some of the static treatment interventions to be compared cannot be followed by all experimental units, due either to baseline characteristics or to the occurrence of certain events over time. For example, the development of adverse effects or contraindications can force a subject to stop an assigned treatment regimen.In this article we propose causal effect models for a user-supplied set of realistic individualized treatment rules. Realistic individualized treatment rules are defined as treatment rules which always map into the set of possible treatment options. Thus, causal effect models for realistic treatment rules do not rely on the ETA assumption and are fully identifiable from the data. Further, these models can be chosen to generalize marginal structural models for static treatment interventions. The estimating function methodology of Robins and Rotnitzky (1992) (analogue to its application in Murphy, et. al. (2001) for a single treatment rule) provides us with the corresponding locally efficient double robust inverse probability of treatment weighted estimator.In addition, we define causal effect models for "intention-to treat" regimens. The proposed intention-to-treat interventions enforce a static intervention until the time point at which the next treatment does not belong to the set of possible treatment options, at which point the intervention is stopped. We provide locally efficient estimators of such intention-to-treat causal effects. PMID- 19122794 TI - Factors associated with sexual abstinence among adolescents in four sub-Saharan African countries. AB - Drawing on nationally representative data collected from Burkinabe, Ghanaian, Malawian, and Ugandan adolescents, this study examines differences among four groups of never married, 15-19 year olds: primary abstainers (sexually inexperienced), secondary abstainers (last sex more than 12 months prior to the survey), recent abstainers (sexually active in last year but not in the last 3 months), and sexually active (had sexual intercourse in the last 3 months). The percentage of primary abstinent adolescents ranged from 42% (Malawian males) to 85% (Ghanaian males). In general, a greater proportion of females than males were primary abstainers. Primary abstainers were younger than sexually experienced adolescents. Current involvement in a romantic relationship was a significant predictor of sexual status with primary abstainers being the least likely to be romantically involved. Overall, findings suggest that adolescents' gender, prior sexual experiences and contextual circumstances, such as romantic partnerships, should be considered when designing abstinence promotion programs. PMID- 19122796 TI - The Teen Marijuana Check-Up: An In-School Protocol for Eliciting Voluntary Self Assessment of Marijuana Use. AB - Given the prevalence of regular marijuana use among adolescents and associated risks for adverse consequences to functioning, effective interventions are needed that are tailored for this population. To date, most such counseling approaches have relied on non-voluntary participation by adolescent marijuana smokers and the outcomes have been only modestly successful. The Teen Marijuana Check-Up is a brief motivational enhancement intervention publicized as a non-pressured and confidential opportunity for the teen marijuana smoker to "take stock" of his/her use. The intervention is designed for in-school implementation and intended to elicit the teen's voluntary participation. This paper highlights the manner in which adolescents are recruited, key intervention elements, the nature of the counseling style utilized, and clinical challenges. The outcomes of two trials with this intervention are briefly discussed. PMID- 19122797 TI - Regioselective Copper-catalyzed C-N and C-S Bond Formation using Amines, Thiols and Halobenzoic Acids. AB - A regioselective method for highly efficient C-N and C-S bond formation with 2 halobenzoic acids is described. The Cu/Cu(2)O-catalyzed reaction is carried out in 2-ethoxyethanol or ethylene glycol diethyl ether and does not require the use of strong base or other additives. This procedure eliminates the need for acid protection, tolerates a wide range of functional groups and provides aromatic and aliphatic amines and sulfides in 81 to 99% yield. PMID- 19122798 TI - Characterization of a bimorph deformable mirror using stroboscopic phase-shifting interferometry. AB - The static and dynamic characteristics of a bimorph deformable mirror (DM) for use in an adaptive optics system are described. The DM is a 35-actuator device composed of two disks of lead magnesium niobate (PMN), an electrostrictive ceramic that produces a mechanical strain in response to an imposed electric field. A custom stroboscopic phase-shifting interferometer was developed to measure the deformation of the mirror in response to applied voltage. The ability of the mirror to replicate optical aberrations described by the Zernike polynomials was tested as a measure of the mirror's static performance. The natural frequencies of the DM were measured up to 20 kHz using both stroboscopic interferometry as well as a commercial laser Doppler vibrometer (LDV). Interferometric measurements of the DM surface profile were analyzed by fitting the surface with mode-shapes predicted using classical plate theory for an elastically supported disk. The measured natural frequencies were found to be in good agreement with the predictions of the theoretical model. PMID- 19122795 TI - Cellular interplay between neurons and glia: toward a comprehensive mechanism for excitotoxic neuronal loss in neurodegeneration. AB - Astrocytes perform vital maintenance, functional enhancement, and protective roles for their associated neurons; however these same mechanisms may become deleterious for neurons under some conditions. In this review, we highlight two normally protective pathways, the endoplasmic reticulum (ER) stress response and an endogenous antioxidant response, which may become neurotoxic when activated in astrocytes during the inflammation associated with neurodegeneration. Stimulation of these multifaceted pathways affects a panoply of cellular processes. Of particular importance is the effect these pathways have on the homeostasis of the excitatory amino acid neurotransmitter, glutamate. The endogenous antioxidant response increases extracellular glutamate in the pursuit of making the cellular antioxidant, glutathione, by increasing expression of the xCT subunit of the cystine/glutamate antiporter. Meanwhile, inflammatory mediators such as TNFalpha reduce levels of membrane-bound glutamate scavenging proteins such as the excitatory amino acid transporters. Together, these cellular activities may result in a net increase in extracellular glutamate that could alter neuronal function and lead to excitotoxicity. Here we discuss the role of N-methyl-D aspartate receptors, which, when excessively stimulated by glutamate, can cause neuronal dysfunction and loss via activation of calpains. While there are other pathways acting in concert or parallel to those we describe here, this review explores a rationale to explain how two protective mechanisms may result in neuronal loss during neurodegeneration. PMID- 19122800 TI - INTERCALATING CONJUGATES OF PEG WITH NUCLEAR LOCALIZATION SIGNAL (NLS) PEPTIDE. PMID- 19122801 TI - Population-Based Assessment of Exposure to Risk Behaviors in Motion Pictures. AB - The aim of most population-based studies of media is to relate a specific exposure to an outcome of interest. A research program has been developed that evaluates exposure to different components of movies in an attempt of assess the association of such exposure with the adoption of substance use during adolescence. To assess exposure to movie substance use, one must measure both viewing time and content. In developing the exposure measure, the study team was interested in circumventing a common problem in exposure measurement, where measures often conflate exposure to media with attention to media. Our aim in this paper is to present a validated measure of exposure to entertainment media, the Beach method, which combines recognition of a movie title with content analysis of the movie for substance use, to generate population based measures of exposure to substance use in this form of entertainment. PMID- 19122799 TI - Heterogeneity in the CD4 T Cell Compartment and the Variability of Neonatal Immune Responsiveness. AB - Over the past decade, it has become clear that T cell immune responses in both murine and human neonates are very heterogeneous, running the gamut from poor or deviant responsiveness to mature, adult-like inflammatory function. How this variability arises is not well understood but there is now a great deal of information suggesting that differences in the T cell compartments in neonates vs adults play important roles. A number of cell types or processes are qualitatively or quantitatively different in the neonate. These include (a) alternate epigenetic programs at the Th2 cytokine locus, (b) enhanced homeostatic proliferation, (c) a relative abundance of fetal-origin cells, (d) a greater representation of recent thymic emigrants, (e) high proportions of potentially self-reactive cells, (f) a developmental delay in the production of regulatory T cells, and (g) cells bearing TCR with limited N region diversity. Different conditions of antigen exposure may lead to different environmental signals that promote the selective responsiveness of one or more of these populations. Therefore, the variability of neonatal responses may be a function of the heterogeneous nature of the responding T cell population. In this review, we will describe these various subpopulations in detail and speculate as to the manner in which they could contribute to the heterogeneity of neonatal immune responses. PMID- 19122802 TI - METHODS FOR AROMATIC AND HETEROCYCLIC AMINE CARCINOGEN-DNA ADDUCT ANALYSIS BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY. AB - Xenobiotic-DNA adducts are used as biomarkers to assess the genotoxic effects of carcinogens. Rats were dosed with 4-aminobiphenyl (4-ABP), 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline (MeIQx), or 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP). DNA was isolated from the colons of vehicle and carcinogen-treated rats and digested using different nucleases and alkaline phosphatase. Deoxyribonucleoside adducts were quantified by capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS) using isotope dilution methods with deuterated internal standards. Major adducts were those bound to the C8 position of deoxyguanosine. 3'- and 5'-Exonucleases were the most efficient nucleases at isolating dG-C8-ABP adducts. However, bulky adducts such as dG-C8 MeIQx and dG-C8-PhIP were better isolated using nuclease P1 rather than a combination of micrococcal nuclease and spleen phosphodiesterase. The use of DNase I enhanced the detection of all three adducts. We describe LC-MS/MS methods for DNA adduct detection and support the testing of different nucleases that increase DNA digestion efficiency and make available more DNA adducts for detection. PMID- 19122805 TI - The Classroom Check-up: A Classwide Teacher Consultation Model for Increasing Praise and Decreasing Disruptive Behavior. AB - School-based consultation typically focuses on individual student problems and on a small number of students rather than on changing the classroom system. The Classroom Check-up (CCU) was developed as a classwide consultation model to address the need for classroom level support while minimizing treatment integrity problems common to school-based consultation. The purpose of the study was to evaluate the effects of the CCU and Visual Performance Feedback on teacher and student behavior. Results indicated that implementation of the CCU plus Visual Performance Feedback increased teacher implementation of classroom management strategies, including increased use of praise, use of behavior specific praise, and decreased use of reprimands. Further, these changes in teacher behavior contributed to decreases in classroom disruptive behavior. The results are encouraging because they suggest that consultation at the classroom level can create meaningful teacher and student behavior change. PMID- 19122804 TI - Putative role of brain acetaldehyde in ethanol addiction. AB - The putative contribution of brain acetaldehyde (AcH) to ethanol (EtOH) tolerance and dependence (addiction) is reviewed. Although the role of AcH in EtOH addiction has been controversial, there are data showing a relationship. AcH can be formed in the brain tissues through the peroxidatic activity of catalase and by oxidation via other oxidizing enzymes such as cytochrome P-4502E1. Significant formation of AcH occurs in vitro in brain tissue at concentrations of EtOH that can be achieved by voluntary consumption of EtOH by rodents. AcH itself possesses reinforcing properties, which suggests that some of the behavioral pharmacological effects attributed to EtOH may be a result of the formation of AcH, and supports the involvement of AcH in EtOH addiction. Modulation of aldehyde dehydrogenase (ALDH) and brain catalase activity can change EtOH-related addictive behaviors presumably by changing AcH levels. Moreover, some condensation reaction products of AcH may promote some actions of EtOH and its consumption. On the basis of the findings, it can be concluded that AcH may mediate some of the CNS actions of EtOH including tolerance and dependence, although further exploration the involvement of AcH in EtOH addiction is warranted. PMID- 19122806 TI - Spaceflight-relevant types of ionizing radiation and cortical bone: Potential LET effect? AB - Extended exposure to microgravity conditions results in significant bone loss. Coupled with radiation exposure, this phenomenon may place astronauts at a greater risk for mission-critical fractures. In a previous study, we identified a profound and prolonged loss of trabecular bone (29-39%) in mice following exposure to an acute, 2 Gy dose of radiation simulating both solar and cosmic sources. However, because skeletal strength depends on trabecular and cortical bone, accurate assessment of strength requires analysis of both bone compartments. The objective of the present study was to examine various properties of cortical bone in mice following exposure to multiple types of spaceflight-relevant radiation. Nine-week old, female C57BL/6 mice were sacrificed 110 days after exposure to a single, whole body, 2 Gy dose of gamma, proton, carbon, or iron radiation. Femora were evaluated with biomechanical testing, microcomputed tomography, quantitative histomorphometry, percent mineral content, and micro-hardness analysis. Compared to non-irradiated controls, there were significant differences compared to carbon or iron radiation for only fracture force, medullary area and mineral content. A greater differential effect based on linear energy transfer (LET) level may be present: high-LET (carbon or iron) particle irradiation was associated with a decline in structural properties (maximum force, fracture force, medullary area, and cortical porosity) and mineral composition compared to low-LET radiation (gamma and proton). Bone loss following irradiation appears to be largely specific to trabecular bone and may indicate unique biological microenvironments and microdosimetry conditions. However, the limited time points examined and non-haversian skeletal structure of the mice employed highlight the need for further investigation. PMID- 19122807 TI - TFIP11, CCNL1 and EWSR1 Protein-protein Interactions, and Their Nuclear Localization. AB - Previous studies using the yeast two-hybrid assay (Y2H) have identified cyclin L1 (CCNL1) and Ewing sarcoma breakpoint region 1 protein (EWSR1) as being interacting partners of tuftelin-interacting protein 11 (TFIP11). All three proteins are functionally related to the spliceosome and involved in pre-mRNA splicing activities. The spliceosome is a dynamic ribonucleoprotein complex responsible for pre-mRNA splicing of intronic regions, and is composed of five small nuclear RNAs (snRNAs) and MU140 proteins. TFIP11 appears to play a role in spliceosome disassembly allowing for the release of the bound lariat-intron. The roles of CCNL1 and EWSR1 in the spliceosome are poorly understood. Using fluorescently-tagged proteins and confocal microscopy we show that TFIP11, CCNL1 and EWSR1 frequently co-localize to speckled nuclear domains. These data would suggest that all three proteins participate in a common cellular activity related to RNA splicing events. PMID- 19122808 TI - Variable Selection in Semiparametric Regression Modeling. AB - In this paper, we are concerned with how to select significant variables in semiparametric modeling. Variable selection for semiparametric regression models consists of two components: model selection for nonparametric components and select significant variables for parametric portion. Thus, it is much more challenging than that for parametric models such as linear models and generalized linear models because traditional variable selection procedures including stepwise regression and the best subset selection require model selection to nonparametric components for each submodel. This leads to very heavy computational burden. In this paper, we propose a class of variable selection procedures for semiparametric regression models using nonconcave penalized likelihood. The newly proposed procedures are distinguished from the traditional ones in that they delete insignificant variables and estimate the coefficients of significant variables simultaneously. This allows us to establish the sampling properties of the resulting estimate. We first establish the rate of convergence of the resulting estimate. With proper choices of penalty functions and regularization parameters, we then establish the asymptotic normality of the resulting estimate, and further demonstrate that the proposed procedures perform as well as an oracle procedure. Semiparametric generalized likelihood ratio test is proposed to select significant variables in the nonparametric component. We investigate the asymptotic behavior of the proposed test and demonstrate its limiting null distribution follows a chi-squared distribution, which is independent of the nuisance parameters. Extensive Monte Carlo simulation studies are conducted to examine the finite sample performance of the proposed variable selection procedures. PMID- 19122809 TI - Asymptotic approximation of an ionic model for cardiac restitution. AB - Cardiac restitution has been described both in terms of ionic models-systems of ODE's-and in terms of mapping models. While the former provide a more fundamental description, the latter are more flexible in trying to fit experimental data. Recently we proposed a two-dimensional mapping that accurately reproduces restitution behavior of a paced cardiac patch, including rate dependence and accommodation. By contrast, with previous models only a qualitative, not a quantitative, fit had been possible. In this paper, a theoretical foundation for the new mapping is established by deriving it as an asymptotic limit of an idealized ionic model. PMID- 19122810 TI - Differential expression of A-type and B-type lamins during hair cycling. AB - Multiple genetic disorders caused by mutations that affect the proteins lamin A and C show strong skin phenotypes. These disorders include the premature aging disorders Hutchinson-Gilford progeria syndrome and mandibuloacral dysplasia, as well as restrictive dermopathy. Prior studies have shown that the lamin A/C and B proteins are expressed in skin, but little is known about their normal expression in the different skin cell-types and during the hair cycle. Our immunohistochemical staining for lamins A/C and B in wild-type mice revealed strong expression in the basal cell layer of the epidermis, the outer root sheath, and the dermal papilla during all stages of the hair cycle. Lower expression of both lamins A/C and B was seen in suprabasal cells of the epidermis, in the hypodermis, and in the bulb of catagen follicles. In addition, we have utilized a previously described mouse model of Hutchinson-Gilford progeria syndrome and show here that the expression of progerin does not result in pronounced effects on hair cycling or the expression of lamin B. PMID- 19122811 TI - Protein hydrolysates are avoided by herbivores but not by omnivores in two-choice preference tests. AB - BACKGROUND: The negative sensory properties of casein hydrolysates (HC) often limit their usage in products intended for human consumption, despite HC being nutritious and having many functional benefits. Recent, but taxonomically limited, evidence suggests that other animals also avoid consuming HC when alternatives exist. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated ingestive responses of five herbivorous species (guinea pig, mountain beaver, gopher, vole, and rabbit) and five omnivorous species (rat, coyote, house mouse, white-footed mouse, and deer mouse; N = 16-18/species) using solid foods containing 20% HC in a series of two-choice preference tests that used a non-protein, cellulose-based alternative. Individuals were also tested with collagen hydrolysate (gelatin; GE) to determine whether it would induce similar ingestive responses to those induced by HC. Despite HC and GE having very different nutritional and sensory qualities, both hydrolysates produced similar preference score patterns. We found that the herbivores generally avoided the hydrolysates while the omnivores consumed them at similar levels to the cellulose diet or, more rarely, preferred them (HC by the white-footed mouse; GE by the rat). Follow-up preference tests pairing HC and the nutritionally equivalent intact casein (C) were performed on the three mouse species and the guinea pigs. For the mice, mean HC preference scores were lower in the HC v C compared to the HC v Cel tests, indicating that HC's sensory qualities negatively affected its consumption. However, responses were species specific. For the guinea pigs, repeated exposure to HC or C (4.7-h sessions; N = 10) were found to increase subsequent HC preference scores in an HC v C preference test, which was interpreted in the light of conservative foraging strategies thought to typify herbivores. CONCLUSIONS/SIGNIFICANCE: This is the first empirical study of dietary niche-related taxonomic differences in ingestive responses to protein hydrolysates using multiple species under comparable conditions. Our results provide a basis for future work in sensory, physiological, and behavioral mechanisms of hydrolysate avoidance and on the potential use of hydrolysates for pest management. PMID- 19122812 TI - Toll-like receptor 3 signaling on macrophages is required for survival following coxsackievirus B4 infection. AB - Toll-like receptor 3 (TLR3) has been proposed to play a central role in the early recognition of viruses by sensing double stranded RNA, a common intermediate of viral replication. However, several reports have demonstrated that TLR3 signaling is either dispensable or even harmful following infection with certain viruses. Here, we asked whether TLR3 plays a role in the response to coxsackievirus B4 (CB4), a prevalent human pathogen that has been associated with pancreatitis, myocarditis and diabetes. We demonstrate that TLR3 signaling on macrophages is critical to establish protective immunity to CB4. TLR3 deficient mice produced reduced pro-inflammatory mediators and are unable to control viral replication at the early stages of infection resulting in severe cardiac damage. Intriguingly, the absence of TLR3 did not affect the activation of several key innate and adaptive cellular effectors. This suggests that in the absence of TLR3 signaling on macrophages, viral replication outpaces the developing adaptive immune response. We further demonstrate that the MyD88-dependent signaling pathways are not only unable to compensate for the loss of TLR3, they are also dispensable in the response to this RNA virus. Our results demonstrate that TLR3 is not simply part of a redundant system of viral recognition, but rather TLR3 plays an essential role in recognizing the molecular signatures associated with specific viruses including CB4. PMID- 19122813 TI - Effective treatment of respiratory alphaherpesvirus infection using RNA interference. AB - BACKGROUND: Equine herpesvirus type 1 (EHV-1), a member of the Alphaherpesvirinae, is spread via nasal secretions and causes respiratory disease, neurological disorders and abortions. The virus is a significant equine pathogen, but current EHV-1 vaccines are only partially protective and effective metaphylactic and therapeutic agents are not available. Small interfering RNAs (siRNA's), delivered intranasally, could prove a valuable alternative for infection control. siRNA's against two essential EHV-1 genes, encoding the viral helicase (Ori) and glycoprotein B, were evaluated for their potential to decrease EHV-1 infection in a mouse model. METHODOLOGY/PRINCIPAL FNDINGS: siRNA therapy in vitro significantly reduced virus production and plaque size. Viral titers were reduced 80-fold with 37.5 pmol of a single siRNA or with as little as 6.25 pmol of each siRNA when used in combination. siRNA therapy in vivo significantly reduced viral replication and clinical signs. Intranasal treatment did not require a transport vehicle and proved effective when given up to 12 h before or after infection. CONCLUSIONS/SIGNIFICANCE: siRNA treatment has potential for both prevention and early treatment of EHV-1 infections. PMID- 19122814 TI - Retrieval of missing spliced leader in dinoflagellates. AB - Spliced leader (SL) trans-splicing has recently been shown to be a common mRNA processing mechanism in dinoflagellates, in which a short (22-nt) sequence, DCCGUAGCCAUUUUGGCUCAAG (D = U, A, or G), is transplanted from the 5'-end of a small non-coding RNA (SL RNA) to the 5' end of mRNA molecules. The widespread existence of the mechanism in dinoflagellates has been demonstrated by detection of this SL (DinoSL) in a wide phylogenetic range of dinoflagellates. Furthermore, the presence of DinoSL in the transcripts of highly diverse groups of nuclear encoded genes has led us to postulate that SL trans-splicing is universal in dinoflagellate nuclear genome. However, some observations inconsistent to this postulation have been reported, exemplified by a recent article reporting apparent absence of DinoSL in the transcripts of some nuclear-encoded genes in Amphidinium carterae. Absence of SL in these gene transcripts would have important implication on gene regulation in dinoflagellates and utility of DinoSL as a universal dinoflagellate-specific primer to study dinoflagellate transcriptomics. In this study, we re-examined transcripts of these genes and found that all of them actually contained DinoSL. Therefore, results to date are consistent to our initial postulation that DinoSL occurs in all dinoflagellate nuclear-encoded mRNAs. PMID- 19122815 TI - Localized populations of CD8 MHC class I tetramer SIV-specific T cells in lymphoid follicles and genital epithelium. AB - CD8 T cells play an important role in controlling viral infections. We investigated the in situ localization of simian immunodeficiency virus (SIV) specific T cells in lymph and genital tissues from SIV-infected macaques using MHC-class I tetramers. The majority of tetramer-binding cells localized in T cell zones and were CD8(+). Curiously, small subpopulations of tetramer-binding cells that had little to no surface CD8 were detected in situ both early and late post infection, and in both vaginally and rectally inoculated macaques. These tetramer(+)CD8(low/-) cells were more often localized in apparent B cell follicles relative to T cell zones and more often found near or within the genital epithelium than the submucosa. Cells analyzed by flow cytometry showed similar populations of cells. Further immunohistological characterization revealed small populations of tetramer(+)CD20(-) cells inside B cell follicles and that tetramer(+) cells did not stain with gammadelta-TCR nor CD4 antibodies. Negative control tetramer staining indicated that tetramer(+)CD8(low/-) cells were not likely NK cells non-specifically binding to MHC tetramers. These findings have important implications for SIV-specific and other antigen-specific T cell function in these specific tissue locations, and suggest a model in which antigen-specific CD8+ T cells down modulate CD8 upon entering B cell follicles or the epithelial layer of tissues, or alternatively a model in which only antigen specific CD8 T cells that down-modulate CD8 can enter B cell follicles or the epithelium. PMID- 19122816 TI - Transmission in heteronymous spinal pathways is modified after stroke and related to motor incoordination. AB - Changes in reflex spinal pathways after stroke have been shown to affect motor activity in agonist and antagonist muscles acting at the same joint. However, only a few studies have evaluated the heteronymous reflex pathways modulating motoneuronal activity at different joints. This study investigates whether there are changes in the spinal facilitatory and inhibitory pathways linking knee to ankle extensors and if such changes may be related to motor deficits after stroke. The early facilitation and later inhibition of soleus H reflex evoked by the stimulation of femoral nerve at 2 times the motor threshold of the quadriceps were assessed in 15 healthy participants and on the paretic and the non-paretic sides of 15 stroke participants. The relationships between this reflex modulation and the levels of motor recovery, coordination and spasticity were then studied. Results show a significant (Mann-Whitney U; P<0.05) increase in both the peak amplitude (mean+/-SEM: 80+/-22% enhancement of the control H reflex) and duration (4.2+/-0.5 ms) of the facilitation on the paretic side of the stroke individuals compared to their non-paretic side (36+/-6% and 2.9+/-0.4 ms) and to the values of the control subjects (33+/-4% and 2.8+/-0.4 ms, respectively). Moreover, the later strong inhibition observed in all control subjects was decreased in the stroke subjects. Both the peak amplitude and the duration of the increased facilitation were inversely correlated (Spearman r = -0.65; P = 0.009 and r = 0.67; P = 0.007, respectively) with the level of coordination (LEMOCOT) of the paretic leg. Duration of this facilitation was also correlated (r = -0.58, P = 0.024) with the level of motor recovery (CMSA). These results confirm changes in transmission in heteronymous spinal pathways that are related to motor deficits after stroke. PMID- 19122817 TI - Increase of universality in human brain during mental imagery from visual perception. AB - BACKGROUND: Different complex systems behave in a similar way near their critical points of phase transitions which leads to an emergence of a universal scaling behaviour. Universality indirectly implies a long-range correlation between constituent subsystems. As the distributed correlated processing is a hallmark of higher complex cognition, I investigated a measure of universality in human brain during perception and mental imagery of complex real-life visual object like visual art. METHODOLOGY/PRINCIPAL FINDINGS: A new method was presented to estimate the strength of hidden universal structure in a multivariate data set. In this study, I investigated this method in the electrical activities (electroencephalogram signals) of human brain during complex cognition. Two broad groups--artists and non-artists--were studied during the encoding (perception) and retrieval (mental imagery) phases of actual paintings. Universal structure was found to be stronger in visual imagery than in visual perception, and this difference was stronger in artists than in non-artists. Further, this effect was found to be largest in the theta band oscillations and over the prefrontal regions bilaterally. CONCLUSIONS/SIGNIFICANCE: Phase transition like dynamics was observed in the electrical activities of human brain during complex cognitive processing, and closeness to phase transition was higher in mental imagery than in real perception. Further, the effect of long-term training on the universal scaling was also demonstrated. PMID- 19122818 TI - Parenting and Child Externalizing Behaviors: Are the Associations Specific or Diffuse? AB - Building upon the link between inadequate parenting and child noncompliance, aggression, and oppositionality, behavioral parent training has been identified as a well-established treatment for externalizing problems in children. Much less empirical attention has been devoted to examining whether inadequate parenting and, in turn, behavioral parent training programs, have specific effects on child externalizing problems or more diffuse effects on both internalizing and externalizing problems. As an initial attempt to examine the specificity of parenting and childhood externalizing problems, this review examines prior research on the association of three parenting behaviors (parental warmth, hostility, and control) with child externalizing versus internalizing problems. Notably, findings revealed relatively little evidence for the specificity of parenting and child externalizing behaviors in the general parenting literature or in the family context of parent depression. Clinical implications and directions for future research are discussed. PMID- 19122819 TI - Factors Influencing Haptic Perception of Complex Shapes. AB - Exploration of an object by arm movement and somatosensation is a serial process that relies on memories and expectations. The present experiments tested the hypothesis that this process involves breaking the object into component shapes (primitives). This was tested by having human subjects explore shapes composed of semicircular arcs, as well as quarter circles or quarter ellipses. The subjects' perception was reported using a visual display. In the first experiment, in which a series of semicircular arcs was presented, with offsets that differed from trial to trial, performance was consistent with the perception of two (left and right) semicircles. In the second experiment, subjects often failed to detect the quarter circles or quarter ellipses and again behaved as if the object was composed of two (top and bottom) semicircles. The results suggest that the synthesis of haptically sensed shapes is biased toward simple geometric objects and that it can be strongly influenced by expectations. PMID- 19122820 TI - The Effect of Pregnancy on Production of Maternal Endogenous Hematopoietic Stem Cells. AB - Fetal microchimerism refers to the presence of fetal cells in maternal blood and tissues during pregnancy. This microchimerism may result from trafficking of fetal and maternal blood across the placenta during pregnancy. Physiological changes in the maternal blood cellular milieu are also recognized during pregnancy and in the early post partum period. Earlier studies showed that maternal blood contains CD34(+) hematopoietic stem cells (HSCs) that bear paternal genetic markers or male phenotype, suggesting that these cells circulated to the mother from male fetuses during pregnancy. Other studies showed that these maternal HSCs have significantly lower expansion potential than their fetal counterparts. We have recently shown increased percentages of CD34(+) HSCs in peripheral blood of pregnant and parous women. Herein, we hypothesize that pregnancy stimulates the production of endogenous CD34(+) HSCs of maternal origin, a phenomenon which potentially could favor post partum regenerative capacity. PMID- 19122821 TI - Survey of u.s. Human research protection organizations: workload and membership. AB - IN A NATIONAL SURVEY OF INSTITUTIONS with federally assured human research protection programs, we obtained workload and other relevant data on their Institutional Review Boards (IRBs) and management organizations. The number of IRBs increased substantially from 1995 to 2005/06 (491 to 2,728 IRBs) with a further increase in 2008 to 3,853 IRBs. Nationally, IRBs reviewed over a quarter million research applications in the year prior to our survey, of which 35% were new applications requiring full committee review. Compared to estimates from 1995, current IRBs review more new and full committee review applications, but the relative percentage of new and full committee applications remained about the same. High volume research institutions have IRBs with a substantially larger per person workload, relative to smaller volume IRBs (i.e., members spent nearly seven times more hours reviewing new applications outside formal committee meetings). Virtually all IRBs included community representatives as members (92%); however, a small number may not be compliant with federal regulations. The present findings suggest the need for research to (a) examine workload and its effects on review quality, research costs, and faculty morale, (b) develop methods for determining optimal fit between IRB workload demands and institutional labor and financing requirements, (c) construct benchmarks for judging reasonable workload for individual IRB members, and (d) examine if the need to recruit IRB scientific expertise from outside the institution, particularly true for smaller research institutions, causes delays in IRB review, and if a more effective way of locating and recruiting experts would improve quality and time to completion. PMID- 19122822 TI - Survey of U.S. Boards that Review Mental Health-related Research. AB - WE OBTAINED DATA ON INSTITUTIONAL Review Boards (IRBs) that review mental health related applications (MHRAs) in a national survey of institutions with federally assured human research protection programs. Approximately 57% of IRBs review MHRAs, and among these a small percentage may not have mental health experts on their committees (5%). Moreover, mental health experts on IRB committees at high research volume institutions are carrying substantially greater workloads than their lower volume counterparts. In terms of committee demographics, more women (36%) are serving as IRB Chairs on committees that review MHRAs than expected from their representation on medical or university faculties; ethnic minority faculty have lower representation among Chairs than might be expected from their overall faculty representation. Our findings suggest the need for additional studies to (a) examine if the number of mental health experts on IRBs should be increased particularly among IRBs reviewing a high volume of MHRAs, (b) determine if the breadth of expertise among IRB mental health experts corresponds to the range of substantive and methodological approaches represented by the mental health protocols under review, and (c) examine if recruiting IRB scientific expertise from outside an institution, a more common practice among smaller research entities, impacts review quality. PMID- 19122823 TI - Is the time dimension of the cell cycle re-entry in AD regulated by centromere cohesion dynamics? AB - Chromosomal involvement is a legitimate, yet not well understood, feature of Alzheimer disease (AD). Firstly, AD affects more women than men. Secondly, the amyloid-beta protein precursor genetic mutations, responsible for a cohort of familial AD cases, reside on chromosome 21, the same chromosome responsible for the developmental disorder Down's syndrome. Thirdly, lymphocytes from AD patients display a novel chromosomal phenotype, namely premature centromere separation (PCS). Other documented morphological phenomena associated with AD include the occurrence of micronuclei, aneuploidy, binucleation, telomere instability, and cell cycle re-entry protein expression. Based on these events, here we present a novel hypothesis that the time dimension of cell cycle re-entry in AD is highly regulated by centromere cohesion dynamics. In view of the fact that neurons can re-enter the cell division cycle, our hypothesis predicts that alterations in the signaling pathway leading to premature cell death in neurons is a consequence of altered regulation of the separation of centromeres as a function of time. It is well known that centromeres in the metaphase-anaphase transition separate in a non-random, sequential order. This sequence has been shown to be deregulated in aging cells, various tumors, syndromes of chromosome instability, following certain chemical inductions, as well as in AD. Over time, premature chromosome separation is both a result of, and a driving force behind, further cohesion impairment, activation of cyclin dependent kinases, and mitotic catastrophe, a vicious circle resulting in cellular degeneration and death. PMID- 19122824 TI - Do Polybrominated Diphenyl Ethers (PBDEs) Increase the Risk of Thyroid Cancer? AB - An increased incidence of thyroid cancer has been reported in many parts of the world including the United States during the past several decades. Recently emerging evidence has demonstrated that polyhalogenated aromatic hydrocarbons (PHAHs), particularly polybrominated diphenyl ethers (PBDEs), alter thyroid hormone homeostasis and cause thyroid dysfunction. However, few studies have been conducted to test whether exposure to PBDEs and other PHAHs increases the risk of thyroid cancer. Here, we hypothesize that elevated exposure to PHAHs, particularly PBDEs, increases the risk of thyroid cancer and may explain part of the increase in incidence of thyroid cancer during the past several decades. In addition, genetic and epigenetic variations in metabolic pathway genes may alter the expression and function of metabolic enzymes which are involved in the metabolism of endogenous thyroid hormones and the detoxification of PBDEs and other PHAHs. Such variation may result in different individual susceptibilities to PBDEs and other PHAHs and the subsequent development of thyroid cancer. The investigation of this hypothesis will lead to an improved understanding of the role of PBDEs and other PHAHs in thyroid tumorigenesis and may provide a real means to prevent this deadly disease. PMID- 19122825 TI - Small ubiquitin-like modifier 4 (SUMO4) polymorphisms and Vogt-Koyanagi-Harada (VKH) syndrome in the Chinese Han population. AB - PURPOSE: To examine whether small ubiquitin-like modifier 4 (SUMO4) polymorphisms were associated with Vogt-Koyanagi-Harada (VKH) syndrome in the Chinese Han population. METHODS: Genotyping for SUMO4 polymorphisms at G-847A, A-504G, A+163G, and C+438T loci was performed on 231 VKH patients and 302 controls using polymerase chain reaction restriction fragment length polymorphism. RESULTS: A decreased frequency of SUMO4 +438 TT genotype was found in VKH patients compared with healthy controls (p=0.009). However, the significance was lost after Bonferroni correction. Human leukocyte antigens (HLA)-DR4 and HLA-DRw53 were significantly associated with susceptibility to VKH syndrome (p=3.21 x 10(-16) and 7.08 x 10(-5), respectively). Stratification analysis based on HLA-DR4 and HLA-DRw53 did not show any associations between SUMO4 polymorphisms and VKH syndrome, although there was a big difference in the percentage of certain allele and genotype frequencies between HLA-DRw53 negative patients and controls. There was no significance in clinical findings and gender stratification analysis. CONCLUSIONS: HLA-DR4 and HLA-DRw53 are strongly associated with the susceptibility to VKH syndrome in the Chinese Han population. However, none of the currently known single nucleotide polymorphisms (SNPs) of SUMO4 are associated with this syndrome. PMID- 19122826 TI - Presence and distribution of 14-3-3 proteins in human ocular surface tissues. AB - PURPOSE: 14-3-3 is a highly conserved, ubiquitously expressed family of proteins. At least seven mammalian isoforms (beta, epsilon, gamma, eta, theta, sigma, and zeta) are known. These proteins associate with over 200 different target molecules and activate several downstream signaling cascades involved in the regulation of metabolism, cell cycle, apoptosis, protein trafficking, transcription, stress responses, and malignant transformations. We are interested in the role of these proteins in the mechanisms regulating homeostasis and the pathologies of the human ocular surface. Therefore, our purpose is to determine the expression of the 14-3-3 proteins in the human cornea, the conjunctiva, and the primary cells comprising these tissues. METHODS: Using immunofluorescence, we determined the expression of 14-3-3 beta, epsilon, gamma, eta, theta, sigma, and zeta in paraffin sections of the human cornea and conjunctiva. Using indirect immunofluorescence and western blot analysis, we also determined the expression of these isoforms in primary corneal epithelial cells, keratocytes, endothelial cells, and primary conjunctival epithelial cells. The expressions of these isoforms in primary epithelial and endothelial cells were compared with the same expressions in several corneal cell lines. Western blot analysis was used to determine the presence of 14-3-3 isoforms in the culture medium from corneal epithelial cells, cell lines, and the tear fluid. RESULTS: All the 14-3-3 isoforms were expressed in the corneal and conjunctival epithelia as well as primary epithelial cells and cell lines. Expression of 14-3-3 sigma was confined to epithelial cells and was secreted into the culture medium of primary cells and cell lines. We also report for the first time that two of the secreted isoforms, 14-3-3 gamma and zeta, are also present in the human tear fluid. CONCLUSIONS: We have determined that all the mammalian 14-3-3 isoforms are expressed in the human cornea, conjunctiva, and the component cells and that the 14-3-3 sigma isoform was found to be epithelial cell specific. We propose that the intracellular and extracellular presence of 14-3-3 sigma suggest its involvement in the epithelia specific signaling pathways. PMID- 19122828 TI - MUC16 expression in Sjogren's syndrome, KCS, and control subjects. AB - PURPOSE: To investigate the expression of MUC16 protein in tears and conjunctival cell membranes and MUC16 mRNA in conjunctival cells of Sjogren's syndrome (SS), keratoconjunctivitus sicca (KCS) and non-dry eyed (NDE) subjects. The relationship of tear flow and soluble MUC16 concentration was also measured. METHODS: Seventy-six subjects were recruited for this study: 25 SS (confirmed via American-European Consensus Criteria 2002), 25 KCS (confirmed by symptoms and Schirmer scores < or =10 mm) and 26 NDE. Tear flow was measured by the Schirmer test without anesthesia for 5 min. Tears were collected using an eye-wash technique. Protein and mRNA were isolated from conjunctival epithelial cells collected via impression cytology. Soluble and membrane bound MUC16 were quantified via western blotting and MUC16 mRNA was quantified by real time qPCR. RESULTS: The SS group demonstrated significantly higher concentrations of soluble MUC16 (7.28 [SS] +/- 3.97 versus 3.35 [KCS] +/- 4.54 [p=0.004] and versus 1.61 [NDE] +/- 1.22 [p<0.001]) and MUC16 mRNA (4.66 [SS] +/- 5.06 versus 1.84 [KCS] +/ 2.26 [p=0.01] and 1.52 [NDE] +/- 1.04 [p=0.003]) compared to both KCS and NDE groups, respectively. No differences in soluble MUC16 or MUC16 mRNA were found between the KCS and NDE groups. Membrane bound MUC16 was similar in all three groups. No significant correlation was found between mean Schirmer values and any measure of MUC16 expression. CONCLUSIONS: These results demonstrate that SS subjects display a significant increase in both soluble MUC16 and MUC16 mRNA concentrations compared to other forms of aqueous deficient dry eye and non dry eyed individuals. There was no correlation between tear flow and soluble MUC16 concentration. PMID- 19122827 TI - Alternate serotype adenovector provides long-term therapeutic gene expression in the eye. AB - PURPOSE: To determine whether the duration of transgene expression from an alternate adenovector serotype, Ad35, can provide advantages over an Ad5 serotype vector following a single intravitreal (IVT) administration. METHODS: To assess the transgene expression profile, mice received one IVT injection of Ad5- or Ad35 based vectors expressing green fluorescent protein (GFP), luciferase or pigment epithelium-derived factor (PEDF). At specified time points following vector administration, eyes were monitored for GFP expression, or eyes were harvested and assayed for adenovector genomes, luciferase activity or PEDF levels. Ad35 based vector in vivo biologic activity was investigated using a mouse model of laser-induced choroidal neovascularization (CNV). On Day 0, mice received one IVT injection of Ad5.PEDF or Ad35.PEDF (HI-RGD) followed by laser-induced CNV on Day 28. Fourteen days later, animals were perfused with fluorescein-labeled dextran and CNV lesion size quantitated in choroidal flat mounts. RESULTS: These studies demonstrate that following a single IVT adenovector administration: 1) gene expression is prolonged following administration of an Ad35 compared to an Ad5 based vector; 2) the amount of vector genomes in the eye remain constant out to 60 days post injection of both Ad5 and Ad35-based vectors; and 3) an Ad35.PEDF (HI-RGD) vector inhibits CNV in a mouse model at 42 days post injection. CONCLUSIONS: These studies show that transgene and genome levels are prolonged in the eye following 1 IVT injection of an Ad35-based vector. Moreover, therapeutic gene levels from 1 IVT administration of Ad35.PEDF (HI-RGD) vector block abnormal blood vessel growth in a laser-induced CNV mouse model. PMID- 19122829 TI - Histone deacetylase inhibitors blocked activation and caused senescence of corneal stromal cells. AB - PURPOSE: Corneal myofibroblasts differentiated from activated corneal stromal cells are the major cellular sources of extracellular matrix synthesis for the repair of corneal injury. In this study, the effects of histone deacetylase (HDAC) inhibitors on the activation, proliferation, migration and senescence of corneal stromal cells were evaluated. METHODS: Primary human and mouse corneal stromal cells were harvested by sequential digestion with dispase and collagenase, and cultured in DMEM/F-12 media under serum-free (keratocytes), serum- (corneal fibroblasts) and TGFbeta1-supplemented (corneal myofibroblasts) conditions. The responses of corneal stromal cells to HDAC inhibitors were characterized by cDNA microarray, real time PCR, immunocytochemistry and western blot analysis. The effects of HDAC inhibitors on corneal fibroblast proliferation, cell cycle distribution, migration and senescence were also assessed in vitro. RESULTS: Fetal bovine serum and TGFbeta1 activated the transdifferentiation of corneal stromal cells into fibroblasts and myofibroblasts, indicated by cell spreading, renewed assembly of actin filaments and enhanced expression of extracellular matrix components, all of which were suppressed by the addition of HDAC inhibitors. HDAC inhibitors inhibited the proliferation of corneal fibroblasts by decreasing the proportion in the S-phase and increasing the proportion in the G0/G1 and G2/M cell cycle checkpoints. HDAC inhibitors showed a dose-dependent inhibitory effects on the migration of corneal fibroblasts. In addition, HDAC inhibitors induced the senescence of corneal myofibroblasts as shown by enhanced staining of beta-galactosidase and upregulated expression of p16(ink4a). CONCLUSIONS: HDAC inhibitors may affect corneal stromal cells by inhibiting myofibroblastic differentiation, cell proliferation, migration and by inducing cell senescence. Thus, this has implications for future studies in the development of promising drugs in the prevention or treatment of corneal haze and scar formation. PMID- 19122830 TI - Linkage analysis of high myopia susceptibility locus in 26 families. AB - PURPOSE: We conducted a linkage analysis in high myopia families to replicate suggestive results from chromosome 7q36 using a model of autosomal dominant inheritance and genetic heterogeneity. We also performed a genome-wide scan to identify novel loci. METHODS: Twenty-six families, with at least two high-myopic subjects (ie. refractive value in the less affected eye of -5 diopters) in each family, were included. Phenotypic examination included standard autorefractometry, ultrasonographic eye length measurement, and clinical confirmation of the non-syndromic character of the refractive disorder. Nine families were collected de novo including 136 available members of whom 34 were highly myopic subjects. Twenty new subjects were added in 5 of the 17 remaining families. A total of 233 subjects were submitted to a genome scan using ABI linkage mapping set LMSv2-MD-10, additional markers in all regions where preliminary LOD scores were greater than 1.5 were used. Multipoint parametric and non-parametric analyses were conducted with the software packages Genehunter 2.0 and Merlin 1.0.1. Two autosomal recessive, two autosomal dominant, and four autosomal additive models were used in the parametric linkage analyses. RESULTS: No linkage was found using the subset of nine newly collected families. Study of the entire population of 26 families with a parametric model did not yield a significant LOD score (>3), even for the previously suggestive locus on 7q36. A non-parametric model demonstrated significant linkage to chromosome 7p15 in the entire population (Z-NPL=4.07, p=0.00002). The interval is 7.81 centiMorgans (cM) between markers D7S2458 and D7S2515. CONCLUSIONS: The significant interval reported here needs confirmation in other cohorts. Among possible susceptibility genes in the interval, certain candidates are likely to be involved in eye growth and development. PMID- 19122831 TI - Analysis of partner of inscuteable (mPins) expression in the developing mouse eye. AB - PURPOSE: Asymmetric cell division (ACD) is the fundamental mechanism underlying the generation of cellular diversity in invertebrates and vertebrates. During Drosophila neuroblast division, this process involves stabilization of the apical complex and interaction between the Inscuteable (Insc) and Partner of inscuteable (Pins) proteins. Both cell-intrinsic factors and cell-cell interactions seem to contribute to cell fate decisions in the retina. The Pins protein is known to play a major role in the asymmetric segregation of cell fate determinants during development of the central nervous system in general, but its role in asymmetric cell divisions and retinoblast cell fate has never been explored. The primary aim of this study was to determine the spatial distribution and time course of mouse homolog of Drosophila Partner of Inscuteable (mPins) expression in the developing and adult mouse eye. METHODS: The expression pattern of mPins was studied in the mouse eye from embryonic (E) stage E11.5 until adulthood, by semiquantitative RT PCR, in situ hybridization, and immunohistochemistry. In addition, variations in mRNA and protein levels for mPins were analyzed in the developing postnatal and adult lens, by semiquantitative RT-PCR, western blot analysis, in situ hybridization, and immunohistochemistry. RESULTS: We detected mPins mRNA at early stages of mouse embryonic eye development, particularly in the neuroblastic layer. In early postnatal development, mPins mRNA was still detected in the neuroblastic layer, but also began to be detectable in the ganglion cell layer. Thereafter, mPins mRNA was found throughout the retina. This pattern was maintained in differentiated adult retina. Immunohistochemical studies showed that mPins protein was present in the neuroblastic layer and the ganglion cell layer during the early stages of postnatal retinal development. At these stages, mPins protein was colocalized with Numb protein, a marker of the ACD. At later postnatal stages, mPins protein was present in all retinal nuclear layers and in the inner plexiform layer. It continued to be detected in these layers in the differentiated retina; the outer plexiform layer and the photoreceptor inner segments also began to display positive immunostaining for mPins. In the adult retina, mPins was also detected in the retinal pigment epithelium and choroidal melanocytes. Throughout development, mPins protein was detected in nonretinal tissues, including the cornea, ciliary body, and lens. We focused our attention on lens development and showed that mPins protein was first detected at E14.5. The most striking results obtained concerned the lens, in which mPins protein distribution switched from the anterior to the posterior region of the lens during embryonic development. Interestingly, in the postnatal and adult lens, mPins protein was detected in all lens cells and fibers. CONCLUSIONS: We provide the first demonstration that mPins protein is expressed from embryonic stages until adulthood in the mouse eye. These results suggest that mPins plays important roles in eye development. This work provides preliminary evidence strongly supporting a role for mPins in the asymmetric division of retinoblasts, and in the structure and functions of adult mouse retina. However, the link between the presence of mPins in different ocular compartments and the possible occurrence of asymmetric cell divisions in these compartments remains to be clarified. Further studies are required to elucidate the in vitro and in vivo functions of mPins in the developing and adult human eye. PMID- 19122832 TI - Glutamate receptor activation triggers OPA1 release and induces apoptotic cell death in ischemic rat retina. AB - PURPOSE: Glutamate receptor activation-induced excitotoxicity has been hypothesized to cause retinal ganglion cell (RGC) death in glaucoma and to link mitochondrial dysfunction in both acute and chronic neurodegenerative disorders. However, the relationships among elevated intraocular pressure (IOP), glutamate receptor-mediated excitotoxicity, and mitochondrial dysfunction in glaucoma remains unknown. The goal of this study was to determine whether the N- methyl D aspartate (NMDA) glutamate receptor antagonist MK801 can block optic atrophy 1 (OPA1) release and subsequent apoptotic cell death, as well as whether acute IOP elevation triggers OPA1 release and alters OPA1 gene and protein expression in the rat retina after ischemia. METHODS: Sprague Dawley rats received injections of MK801 (10 mg/kg) or vehicle and then transient retinal ischemia was induced by acute IOP elevation. Following subcellular fractionation, changes in cytoplasmic and mitochondrial OPA1 were assessed by western blot analysis. Also, the expression of OPA1 mRNA was measured by Taqman qPCR, the distribution of OPA1 protein was assessed by immunohistochemistry, and apoptotic cell death was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. RESULTS: The ~65 and 90 kDa isoforms of OPA1 were increased in the cytosol in the rat retina at 6 h and at 12 h, but only the 90 kDa isoform of OPA1 was decreased at 12 h after ischemia induced by acute IOP elevation. This suggests that ischemic insult induced OPA1 release from the mitochondria in retinas. Pretreatment with MK801 blocked this effect and significantly increased OPA1 immunoreactivity in the inner retinal layers, as well as OPA1 gene expression and total protein expression in retinas at 12 h after ischemia. Further, pretreatment with MK801 prevented apoptotic cell death in retinas at 12 h after ischemia. Following acute IOP elevation, Bcl-2 mRNA expression in retinas was decreased at 3 h and 6 h but increased at 12 h and 24 h. In contrast, Bax mRNA expression in these retinas was increased in the first 12 h and then plateaued. Moreover, pretreatment with MK801 increased Bcl-2 mRNA expression, but did not alter the course of Bax mRNA expression. CONCLUSIONS: These results indicate that OPA1 release from mitochondria triggered by acute IOP elevation is inhibited by blockade of glutamate receptor activation. Because this effect was accompanied by increases of Bcl-2 expression, no changes of Bax expression, and blockade of apoptosis, these findings indicate that glutamate receptor activation following acute IOP elevation may lead to a distinct mitochondria-mediated cell death pathway in ischemic retina. These results support further studies to determine whether ischemia-induced OPA1 release may be an important component of the biochemical cascade leading to pressure-related ischemic damage in glaucomatous retina. PMID- 19122833 TI - ARHI: A new target of galactose toxicity in Classic Galactosemia. AB - In humans, deficiency of galactose-1-phosphate uridyltransferase (GALT) activity can lead to a potentially lethal disease called Classic Galactosemia. Although a galactose-restricted diet can prevent the acute lethality associated with the disorder, chronic complications persist in many well-treated patients. Approximately 85% of young women with Classic Galactosemia experience hypergonadotropic hypogonadism and premature ovarian failure (POF). Others suffer from mental retardation, growth restriction, speech dyspraxia, and ataxia. Despite decades of intense biochemical characterization, little is known about the molecular etiology, as well as the chronology of the pathological events leading to the poor outcomes. Several hypotheses have been proposed, most of which involved the accumulation of the intermediates and/or the deficit of the products, of the blocked GALT pathway. However, none of these hypotheses satisfactorily explained the absence of patient phenotypes in the GALT-knockout mice. Here we proposed that the gene encoded the human tumor suppressor gene aplysia rashomolog I (ARHI) is a target of toxicity in Classic Galactosemia, and because ARHI gene is lost in rodents in through evolution, it thus accounts for the lack of clinical phenotypes in the GALT-knockout mice. PMID- 19122834 TI - Human Brain Imaging of Tinnitus and Animal Models. AB - Because subjective tinnitus is typically localized to the ear with hearing loss, tinnitus was traditionally thought to originate from neural hyperactivity in the damaged ear. However, most studies have found that hearing loss reduces the neural outputs from the damaged cochlea. These negative findings led to the hypothesis that rinnitus arises from aberrant neural activity in the central auditory system. Positron emission tomography imaging studies performed on tinnitus patients that could modulate their tinnitus provide evidence showing that the aberrant neural activity that gives rise to tinnitus resides in the central auditory pathway. To investigate the biological basis of tinnitus in more detail, an animal model was developed that allowed behavioral measures of tinnitus to be obtained from individual rats after inducing tinnitus with high doses of salicylate or high-intensity noise. This behavioral model was used to test the efficacy of memantine, an N-methyl-D-aspartate antagonist, and scopolamine, an anticholinergic, in suppressing salicylate-induced tinnitus. Neither drug completely suppressed salicylate-induced tinnitus. To detect the physiological changes associated with tinnitus, chronic microwire electrodes were implanted in the auditory cortex and measurements were obtained from the auditory cortex before and after salicylate and noise exposures known to induce tinnitus. High doses of salicylate or high-level noise exposure generally resulted in sound evoked hyperactivity in the electrophysiological responses recorded from the auditory cortex of awake-animals. However, anesthetic tended to suppress or abolish the hyperactivity. PMID- 19122835 TI - Educating osteopaths to be researchers - what role should research methods and statistics have in an undergraduate curriculum? AB - Evidence-based medicine (EBM) involves using research data to enhance the diagnosis and treatment of clinical disorders. Somatic dysfunction and osteopathic manipulative treatment (OMT) are two unique aspects of osteopathy that will benefit from a greater emphasis on scientific evidence. Most evidence in osteopathy is based on expert opinions, case reports, case series, and observational studies. Only one systematic review of randomized controlled trials, involving OMT for low back pain, has been published. Although this study demonstrates the efficacy of OMT for low back pain, other clinical trials are needed to expand the evidence base in osteopathy. Undergraduate osteopathy curricula should ensure that students acquire the tools necessary to become knowledgeable consumers of the research and statistics presented in biomedical journals. Such curricula need to be supplemented with graduate training programs and research funding mechanisms to ensure that young osteopathic researchers are able to produce the research needed to practice and advance evidence-based osteopathy in the future. PMID- 19122837 TI - Racial-ethnic self-schemas: Multi-dimensional identity-based motivation. AB - Prior self-schema research focuses on benefits of being schematic vs. aschematic in stereotyped domains. The current studies build on this work, examining racial ethnic self-schemas as multi-dimensional, containing multiple, conflicting, and non-integrated images. A multidimensional perspective captures complexity; examining net effects of dimensions predicts within-group differences in academic engagement and well-being. When racial-ethnicity self-schemas focus attention on membership in both in-group and broader society, engagement with school should increase since school is not seen as out-group defining. When racial-ethnicity self-schemas focus attention on inclusion (not obstacles to inclusion) in broader society, risk of depressive symptoms should decrease. Support for these hypotheses was found in two separate samples (8(th) graders, n = 213, 9(th) graders followed to 12(th) grade n = 141). PMID- 19122838 TI - "The Great Unmentionable": Exploring the Pleasures and Benefits of Ecstasy from the Perspectives of Drug Users. PMID- 19122839 TI - Culture sometimes matters: intra-cultural variation in pro-social behavior among Tsimane Amerindians. AB - Agent-centered models usually consider only individual-level variables in calculations of economic costs and benefits. There has been little consideration of social or cultural history on shaping payoffs in ways that impact decisions. To examine the role of local expectations on economic behavior, we explore whether village affiliation accounts for the variation in Dictator Game offers among the Tsimane of the Bolivian Amazon independently of other factors that could confound such an effect. Our analysis shows that significant differences in altruistic giving exist among villages, village patterns are recognized by residents, and offers likely reflect variation in social expectations rather than stable differences in norms of fairness. PMID- 19122840 TI - INTERSPECIES HYBRIDIZATION AND THE STRATIFICATION OF NUCLEAR GENETIC VARIATION OF RHESUS (MACACA MULATTA) AND LONG-TAILED MACAQUES (MACACA FASCICULARIS). AB - Genotypes for 13 short tandem repeats (STRs) were used to assess the genetic diversity within and differentiation among populations of rhesus macaques (Macaca mulatta) from mainland Asia and long-tailed macaques (M. fascicularis) from mainland and insular Southeast Asia. These animals were either recently captured in the wild or derived from wild-caught founders maintained in captivity for biomedical research.A large number of alleles is shared between the two macaque species but a significant genetic division between them persists. This distinction is more clear-cut among populations that are not, or are unlikely to have recently been, geographically contiguous. Our results suggest there has been significant interspecies nuclear gene flow between rhesus macaques and long tailed macaques on the mainland. Comparisons of mainland and island populations of long-tailed macaques reflect marked genetic subdivisions due to barriers to migration. Geographic isolation has restricted gene flow, allowing island populations to become subdivided and genetically differentiated. Indonesian long tailed macaques show evidence of long-term separation and genetic isolation from the mainland populations, while long-tailed macaques from the Philippines and Mauritius both display evidence of founder effects and subsequent isolation, with the impact from genetic drift being more profound in the latter. PMID- 19122841 TI - A Colorimetric Chemodosimeter for Pd(II): A Method for Detecting Residual Palladium in Cross-Coupling Reactions. AB - A colorimetric chemodosimeter (SQ1) for the detection of trace palladium salts in cross-coupling reactions mediated by palladium is described. Decolorization of SQ1 is affected by nucleophilic attack of ethanethiol in basic DMSO solutions. Thiol addition is determined to have an equilibrium constant (K(eq)) of 2.9 * 10(6) M(-1), with a large entropic and modest enthalpic driving force. This unusual result is attributed to solvent effects arising from a strong coordinative interaction between DMSO and the parent squaraine. Palladium detection is achieved through thiol scavenging from the SQ1-ethanethiol complex leading to a color "turn-on" of the parent squaraine. It was found that untreated samples obtained directly from Suzuki couplings showed no response to the assay. However, treatment of the samples with aqueous nitric acid generates a uniform Pd(NO(3))(2) species, which gives an appropriate response. "Naked-eye" detection of Pd(NO(3))(2) was estimated to be as low as 0.5 ppm in solution, and instrument based detection was tested as low as 100 ppb. The average error over the working range of the assay was determined to be 7%. PMID- 19122842 TI - Functionalized carbon nanotubes and nanofibers for biosensing applications. AB - This review summarizes recent advances in electrochemical biosensors based on carbon nanotubes (CNTs) and carbon nanofibers (CNFs) with an emphasis on applications of CNTs. CNTs and CNFs have unique electric, electrocatalytic and mechanical properties, which make them efficient materials for developing electrochemical biosensors.We discuss functionalizing CNTs for biosensors. We review electrochemical biosensors based on CNTs and their various applications (e.g., measurement of small biological molecules and environmental pollutants, detection of DNA, and immunosensing of disease biomarkers). Moreover, we outline the development of electrochemical biosensors based on CNFs and their applications. Finally, we discuss some future applications of CNTs. PMID- 19122843 TI - Synthesis of Co/MFe(2)O(4) (M = Fe, Mn) Core/Shell Nanocomposite Particles. AB - Monodispersed cobalt nanoparticles (NPs) with controllable size (8-14 nm) have been synthesized using thermal decomposition of dicobaltoctacarbonyl in organic solvent. The as-synthesized high magnetic moment (125 emu/g) Co NPs are dispersible in various organic solvents, and can be easily transferred into aqueous phase by surface modification using phospholipids. However, the modified hydrophilic Co NPs are not stable as they are quickly oxidized, agglomerated in buffer. Co NPs are stabilized by coating the MFe(2)O(4) (M = Fe, Mn) ferrite shell. Core/shell structured bimagnetic Co/MFe(2)O(4) nanocomposites are prepared with tunable shell thickness (1-5 nm). The Co/MFe(2)O(4) nanocomposites retain the high magnetic moment density from the Co core, while gaining chemical and magnetic stability from the ferrite shell. Comparing to Co NPs, the nanocomposites show much enhanced stability in buffer solution at elevated temperatures, making them promising for biomedical applications. PMID- 19122836 TI - Peptide-based Biopolymers in Biomedicine and Biotechnology. AB - Peptides are emerging as a new class of biomaterials due to their unique chemical, physical, and biological properties. The development of peptide-based biomaterials is driven by the convergence of protein engineering and macromolecular self-assembly. This review covers the basic principles, applications, and prospects of peptide-based biomaterials. We focus on both chemically synthesized and genetically encoded peptides, including poly-amino acids, elastin-like polypeptides, silk-like polymers and other biopolymers based on repetitive peptide motifs. Applications of these engineered biomolecules in protein purification, controlled drug delivery, tissue engineering, and biosurface engineering are discussed. PMID- 19122844 TI - 2',2'-Ligation Demonstrates the Thermal Dependence of DNA-Directed Positional Control. AB - An oligodeoxynucleotide (ODN) containing a 2'-aminoalkylmercaptouridine was ligated chemically with another ODN containing a 2'-carboxyalkylmercaptouridine to produce a 2',2'-amide linked conjugate. Templation by a DNA hairpin resulted in ligation yields of > 95%. At room temperature, similar yields were obtained for amide ligations between pendent nucleotides located adjacent to each other or separated by three nucleotides. However, at 4 degrees C, ligation was vastly favored for the case where the pendent amine and carboxylate were attached to adjacent residues (i.e., at the 3'- and 5'-termini of the two ODNs). No reaction was observed to occur either at 4 degrees C or at room temperature between pendent nucleotides separated by 9 nucleotides. Denaturing gel electrophoresis, MALDI-TOF MS and full nuclease digestion coupled with LC-MS were used to characterize the reaction products. PMID- 19122845 TI - An examination of the association between the 5-HTT promoter region polymorphism and depressogenic attributional styles in childhood. AB - Although a vast literature examining the role of attributional styles in depression has accumulated, the origins of such cognitions remain poorly understood. Investigators are increasingly interested in whether cognitive vulnerability to depression is linked to genetic variation. As a preliminary test of this hypothesis, we examined whether the serotonin transporter promoter polymorphism (5-HTTLPR) was associated with attributional styles in children. Thirty-eight children completed a self-report measure of attributional styles, the Child Attributional Style Questionnaire-Revised (CASQ-R). Children were also genotyped for the 5-HTTLPR polymorphism, including the single nucleotide polymorphism (SNP) rs25531 in the long allele of the 5-HTTLPR. The short alleles of the 5-HTTLPR and their putative functional equivalents were associated with increased levels of depressogenic attributions for negative events, as measured by the CASQ-R, lending support to the role of 5-HTTLPR polymorphisms in cognitive vulnerability to depression. PMID- 19122846 TI - Using Influenza-Like Illness Data to Reconstruct an Influenza Outbreak. AB - The objective of this study was to reconstruct the type A influenza epidemic that occurred in the Research Triangle Park (RTP) region of North Carolina during the 2003-04 flu season. We describe an agent-based influenza transmission model that uses Influenza-like Illness (ILI) data gathered from state agencies to estimate model parameters. The design of the model is similar to models represented in the literature that have been used to predict the impact of pandemic avian influenza in Southeast Asia and in the continental United States and to assess containment strategies. The focus of this model aims to reconstruct a historical epidemic that left traces of its impact in the form of an ILI epidemic curve. In this context, the work assumes aspects of a curve fitting exercise. PMID- 19122848 TI - Studies on Taxol Biosynthesis: Preparation of Taxadiene-diol- and triol Derivatives by Deoxygenation of Taxusin. AB - The putative taxol biosynthesis metabolites, taxa-4(20),11(12)-diene-5alpha, 13alpha -diol (7), taxa-4(20),11(12)-diene-5alpha, 9alpha, 13alpha-triol (9), and taxa-4(20),11(12)-diene-5alpha, 10beta, 13alpha-triol (10), have been prepared by Barton deoxygenation of the C-9 and C10-hydroxyl groups of protected derivatives of taxusin, a major taxoid metabolite isolated from Yew heart wood. The synthetic protocol devised, is amenable for the preparation of isotopically labeled congeners that will be useful to probe further intermediate steps in the biosynthesis of taxol. PMID- 19122847 TI - Functional effects of KCNE3 mutation and its role in the development of Brugada syndrome. AB - INTRODUCTION: The Brugada Syndrome (BrS), an inherited syndrome associated with a high incidence of sudden cardiac arrest, has been linked to mutations in four different genes leading to a loss of function in sodium and calcium channel activity. Although the transient outward current (I(to)) is thought to play a prominent role in the expression of the syndrome, mutations in I(to)-related genes have not been identified as yet. METHODS AND RESULTS: One hundred and five probands with BrS were screened for ion channel gene mutations using single strand conformation polymorphism (SSCP) electrophoresis and direct sequencing. A missense mutation (R99H) in KCNE3 (MiRP2) was detected in one proband. The R99H mutation was found 4/4 phenotype positive and 0/3 phenotype-negative family members. Chinese hamster ovary (CHO)-K1 cells were co-transfected using wild-type (WT) or mutant KCNE3 and either WT KCND3 or KCNQ1. Whole-cell patch clamp studies were performed after 48 hours. Interactions between Kv4.3 and KCNE3 were analyzed in co-immunoprecipitation experiments in human atrial samples. Co-transfection of R99H-KCNE3 with KCNQ1 produced no alteration in current magnitude or kinetics. However, co-transfection of R99H KCNE3 with KCND3 resulted in a significant increase in the I(to) intensity compared to WT KCNE3+KCND3. Using tissues isolated from left atrial appendages of human hearts, we also demonstrate that K(v)4.3 and KCNE3 can be co-immunoprecipitated. CONCLUSIONS: These results provide definitive evidence for a functional role of KCNE3 in the modulation of I(to) in the human heart and suggest that mutations in KCNE3 can underlie the development of BrS. PMID- 19122849 TI - Ethnicity, Education, and the Temporal Stability of Personality Traits In the East Baltimore Epidemiologic Catchment Area Study. AB - We examined the influence of age, gender, Black vs. White ethnicity, and education on five indices of personality stability and change across an average interval of 8 years in the East Baltimore Epidemiologic Catchment Area study. In the full sample (n = 505, aged 30-88), examination of structural, rank-order, ipsative, and mean level stability, as well as indices of reliable change suggested that NEO-PI-R personality traits showed moderate to high levels of stability over time. There were few age and gender effects on temporal stability but rank-order, ipsative, and mean level stability were lower among Blacks and individuals with lower education. Future research should explore additional demographic predictors of temporal plasticity in a diverse range of samples, and employ observer ratings to assess personality. PMID- 19122850 TI - Fearful Temperament and Stress Reactivity Among Preschool-Aged Children. AB - In this study, we examined the relation between physiological stress-reactivity and temperamental fearfulness in 162 preschool-aged children. Both the autonomic and neuroendocrine arms of the mammalian stress system were examined. Larger stress responses were defined as greater sympathetic activation, parasympathetic withdrawal and cortisol increases to stressor tasks. Fearful temperament was examined using parent report and behavior in response to fear-evocative laboratory tasks. There was little evidence that larger sympathetic activation or parasympathetic withdrawal was associated with fearful temperament. Greater cortisol reactivity, however, was associated with fearful temperament. Additional analyses examined those children who were consistently fearful across all measures, and the results remained largely the same. However, there was some suggestion that consistently fearful compared to non-fearful children might be more likely to exhibit sympathetic activation to the fear-evocative stimuli. These findings provide support for the argument that fearful temperament is associated with greater stress reactivity in young children. Nonetheless the size of the associations was small and future studies will need to determine whether reactivity of stress-sensitive physiological systems contributes to the development of individual differences in fearful temperament or merely reflects these differences. PMID- 19122851 TI - Personality in Middle Childhood: A Hierarchical Structure and Longitudinal Connections With Personality in Late Adolescence. AB - Research on the structure of personality in middle childhood, while advancing, is still in the early stages of development. In this study, we employed a group of 1563 twins to elucidate the hierarchical structure of personality in middle childhood and provide connections to established personality traits in adult populations. Our results provide evidence for a higher-order structure of personality in middle childhood that maps on to recent findings in adult populations supporting hierarchical relationships among 2-, 3-, 4-, and 5-factor models of personality. In addition, primary higher-order personality traits rated by parents at age 11 showed substantial predictive validity for analogous traits rated by self at age 17. We discuss our results within the context of developing a convergent hierarchical taxonomy of personality in middle childhood and the importance of multiinformant investigations. PMID- 19122852 TI - Protein-passivated Fe(3)O(4) nanoparticles: low toxicity and rapid heating for thermal therapy. AB - Thermotherapy is a promising technique for the minimally invasive elimination of solid tumors. Here we report the fabrication of protein-coated iron oxide NPs (12 nm core) for use as thermal therapeutic agents. These albumin-passivated NPs are stable under physiological conditions, with rapid heating and cell killing capacity upon alternating magnetic field (AMF) exposure. The mode of action is specific: no measurable cytotoxicity was observed for the particle without AMF or for AMF exposure without the particle. PMID- 19122853 TI - The Role of Representational Status and Item Complexity in Parent-Child Conversations about Pictures and Objects. AB - Mother-child conversations about pictures systematically differ from mother-child conversations about objects: Pictures are more likely than objects to elicit talk about kinds, whereas objects are more likely than pictures to elicit talk about individuals. The purpose of the current study is to examine whether this difference between pictures and objects is explained by differences in item complexity. Mothers and their 4-year-old children were randomly assigned to one of two conditions: Simple or Complex. In each condition, participants viewed 12 toy objects and 12 pictures, matched for content. The items were either highly detailed (complex condition) or very plain (simple condition). Replicating previous research, mothers and children provided relatively more focus on kinds when talking about pictures, and relatively more focus on individuals when talking about objects. The current results go further, however, to demonstrate that this effect is independent of the items' complexity. We therefore propose that the picture-object difference is not due to low-level differences in amount of perceptual detail provided, but rather is due to the greater ease with which pictures serve as representations (DeLoache, 1991). These data indicate the ways in which a fundamental conceptual distinction between kinds and individuals arises in different linguistic expressions and in different contexts. PMID- 19122854 TI - Nanoscale potentiometry. AB - Potentiometric sensors share unique characteristics that set them apart from other electrochemical sensors. Potentiometric nanoelectrodes have been reported and successfully used for many decades, and we review these developments. Current research chiefly focuses on nanoscale films at the outer or the inner side of the membrane, with outer layers for increasing biocompatibility, expanding the sensor response, or improving the limit of detection (LOD). Inner layers are mainly used for stabilizing the response and eliminating inner aqueous contacts or undesired nanoscale layers of water. We also discuss the ultimate detectability of ions with such sensors and the power of coupling the ultra-low LODs of ion-selective electrodes with nanoparticle labels to give attractive bioassays that can compete with state-of-the-art electrochemical detection. PMID- 19122855 TI - Long-term High Salt Diet Causes Hypertension and Decreases Renal Expression of Vascular Endothelial Growth Factor in Sprague-Dawley Rats. AB - We seek to determine: 1) whether a long-term high salt diet induces hypertension and renal injury in Sprague-Dawley (SD) rats and 2) whether the high salt diet induced hypertension and renal injury are associated with decreased renal VEGF expression. Twelve 10-wk-old male SD rats received a high salt diet (HS, 8%) and twelve SD rats received a normal salt diet (NS, 0.5 %) for 8 weeks. Using a tail cuff, weekly monitoring showed that blood pressure increased significantly after 6, 7, & 8 wks in HS group, compared to NS group (P<0.01). At 4 wks and 8 wks of diet, mean arterial pressure (MAP) was determined in conscious rats by continuous monitoring through a catheter placed in the carotid artery. MAP was not significantly different between HS and NS group in 4 wks, but was significantly higher in HS than NS group (140+/-5.3 vs.112+/-2.2 mmHg; P<0.01) in 8 wks. Increased proteinuria and albuminuria were associated with marked renal histological abnormalities in HS group, compared to those in NS group. Northern blot and ELISA demonstrated that 8 wks of HS diet significantly decreased renal expression of VEGF mRNA and protein, compared to NS group (P<0.01). In 8 wks, total urinary excretion of sFlt-1 was significantly higher in HS than NS group (9.28+/-1.05 vs. 2.05+/-0.55 ng/day; P<0.01) whereas the plasma levels of sFlt-1 remained stable. These results suggest that a long-term HS diet induces renal injury and hypertension, which are associated with decreased renal VEGF expression in normotensive rodent animals. PMID- 19122856 TI - Stereotypy in Autism: The Importance of Function. AB - We argue for the utility of a functional definition of stereotypy based on evidence of both sensory automatic and socially mediated reinforcement contingencies in the occurrence of stereotypy in children with autism. A predetermined sensory function of stereotypy is often invoked in the behavioral literature and the term "self-stimulatory behavior" is commonly misused as interchangeable with "stereotypy." We discuss evidence for a variety of potential functional properties of stereotypy. Diagnostic definitions are reviewed and support for both sensory and social functions is outlined. We argue that stereotypies should be described and categorized according to their function, rather than form. Furthermore, treatment decisions should be based on a functional interpretation of stereotypy, which acknowledges its operant and heterogeneous quality in autism. PMID- 19122857 TI - Automatic Prosodic Event Detection Using Acoustic, Lexical, and Syntactic Evidence. AB - With the advent of prosody annotation standards such as tones and break indices (ToBI), speech technologists and linguists alike have been interested in automatically detecting prosodic events in speech. This is because the prosodic tier provides an additional layer of information over the short-term segment level features and lexical representation of an utterance. As the prosody of an utterance is closely tied to its syntactic and semantic content in addition to its lexical content, knowledge of the prosodic events within and across utterances can assist spoken language applications such as automatic speech recognition and translation. On the other hand, corpora annotated with prosodic events are useful for building natural-sounding speech synthesizers. In this paper, we build an automatic detector and classifier for prosodic events in American English, based on their acoustic, lexical, and syntactic correlates. Following previous work in this area, we focus on accent (prominence, or "stress") and prosodic phrase boundary detection at the syllable level. Our experiments achieved a performance rate of 86.75% agreement on the accent detection task, and 91.61% agreement on the phrase boundary detection task on the Boston University Radio News Corpus. PMID- 19122858 TI - Modeling of the transient responses of the vocal fold lamina propria. AB - The human voice is produced by flow-induced self-sustained oscillation of the vocal fold lamina propria. The mechanical properties of vocal fold tissues are important for understanding phonation, including the time-dependent and transient changes in fundamental frequency (F(0)). Cyclic uniaxial tensile tests were conducted on a group of specimens of the vocal fold lamina propria, including the superficial layer (vocal fold cover) (5 male, 5 female) and the deeper layers (vocal ligament) (6 male, 6 female). Results showed that the vocal fold lamina propria, like many other soft tissues, exhibits both elastic and viscous behavior. Specifically, the transient mechanical responses of cyclic stress relaxation and creep were observed. A three-network constitutive model composed of a hyperelastic equilibrium network in parallel with two viscoplastic time dependent networks proves effective in characterizing the cyclic stress relaxation and creep behavior. For male vocal folds at a stretch of 1.4, significantly higher peak stress was found in the vocal ligament than in the vocal fold cover. Also, the male vocal ligament was significantly stiffer than the female vocal ligament. Our findings may help explain the mechanisms of some widely observed transient phenomena in F(0) regulation during phonation, such as the global declination in F(0) during the production of declarative sentences, and local F(0) changes such as overshoot and undershoot. PMID- 19122859 TI - Development of analytical methods for multiplex bio-assay with inductively coupled plasma mass spectrometry. AB - Advances in the development of highly multiplexed bio-analytical assays with inductively coupled plasma mass spectrometry (ICP-MS) detection are discussed. Use of novel reagents specifically designed for immunological methods utilizing elemental analysis is presented. The major steps of method development, including selection of elements for tags, validation of tagged reagents, and examples of multiplexed assays, are considered in detail. The paper further describes experimental protocols for elemental tagging of antibodies, immunostaining of live and fixed human leukemia cells, and preparation of samples for ICP-MS analysis. Quantitative analysis of surface antigens on model cell lines using a cocktail of seven lanthanide labeled antibodies demonstrated high specificity and concordance with conventional immunophenotyping. PMID- 19122860 TI - Providing Spatial Data for Secondary Analysis: Issues and Current Practices relating to Confidentiality. AB - Spatially explicit data pose a series of opportunities and challenges for all the actors involved in providing data for long-term preservation and secondary analysis -- the data producer, the data archive, and the data user. We report on opportunities and challenges for each of the three players, and then turn to a summary of current thinking about how best to prepare, archive, disseminate, and make use of social science data that have spatially explicit identification. The core issue that runs through the paper is the risk of the disclosure of the identity of respondents. If we know where they live, where they work, or where they own property, it is possible to find out who they are. Those involved in collecting, archiving, and using data need to be aware of the risks of disclosure and become familiar with best practices to avoid disclosures that will be harmful to respondents. PMID- 19122861 TI - The evolving role of nuclear molecular imaging in cancer. AB - BACKGROUND: Novel therapies targeted to specific tumor pathways are entering the clinic. The need for in vivo monitoring of resulting molecular changes, particularly with respect to the tumor microenvironment, is growing. Molecular imaging is evolving to include a variety of imaging methods to enable in vivo monitoring of cellular and molecular processes. OBJECTIVES: This article reviews the emerging role of molecular imaging in the development of improved therapeutic strategies that provide better patient selection for therapeutic personalization (i.e. determine which therapies have the greatest chance of success given the individual patient's disease genetic, and phenotypical profile). METHODS: In order to illustrate the utility of integrating molecular imaging into therapy development strategies, current and emerging applications of nuclear molecular imaging strategies will be compared with conventional strategies. Proposed methods of integrating molecular imaging techniques into cancer therapeutic development and limitations of these techniques will be discussed. RESULTS/CONCLUSION: Molecular imaging provides a variety of new tools to accelerate the development of cancer therapies. The recent drive to develop molecular imaging probes and standardize molecular imaging techniques is creating the scaffolding for the evolving paradigm shift to personalized cancer therapy. PMID- 19122862 TI - The moderating effects of religiosity on the relationship between stressful life events and delinquent behavior. AB - Previous research has shown that many forms of strain are positively related to delinquency. Evidence also suggests that religiosity buffers the effects of strain on offending, but this issue requires further research. Using data from a national sample of adolescents, this study examined whether or not religiosity conditioned the relationship between strain and delinquency. This study also looked at the ability of social support, self-esteem, and depression to moderate the influence of strain on delinquent behavior. The findings here lend support to general strain theory in that strain had a direct positive effect on delinquency, yet there was little evidence that the relationship was moderated by religiosity or other conditioning variables. The roles of moderating variables on strain across genders were also considered. PMID- 19122864 TI - Commentary on "My Story: A Genetic Counselor's Journey from Provider to Patient" PMID- 19122867 TI - A geometric approach to spectral subtraction. AB - The traditional power spectral subtraction algorithm is computationally simple to implement but suffers from musical noise distortion. In addition, the subtractive rules are based on incorrect assumptions about the cross terms being zero. A new geometric approach to spectral subtraction is proposed in the present paper that addresses these shortcomings of the spectral subtraction algorithm. A method for estimating the cross terms involving the phase differences between the noisy (and clean) signals and noise is proposed. Analysis of the gain function of the proposed algorithm indicated that it possesses similar properties as the traditional MMSE algorithm. Objective evaluation of the proposed algorithm showed that it performed significantly better than the traditional spectral subtractive algorithm. Informal listening tests revealed that the proposed algorithm had no audible musical noise. PMID- 19122866 TI - The Impact of Foster Care and Temporary Assistance for Needy Families (TANF) on Women's Drug Treatment Outcomes. AB - This study assesses the impact of having a child in foster care and receiving cash benefits through Temporary Assistance for Needy Families (TANF) on women's completion of a residential drug treatment program. The study's hypothesis was that drug treatment completion rates for women who had children in foster care and/or who were receiving TANF would differ from women who did not receive these services. The sample included 117 women age 19 to 54, in a Midwestern state. Findings suggest that women with a child or children in foster care were less likely to complete treatment. Women receiving cash benefits were also somewhat less likely to complete treatment than women not receiving these services. Women with children in foster care had similar levels of psychological, employment, and drug and alcohol concerns as other women, as measured by the Addiction Severity Index. Future research should focus on identifying strategies that enhance retention rates of these vulnerable women. Implications for improving treatment retention are discussed in light of the Adoption and Safe Families Act of 1997 and the Personal Responsibility and Work Opportunity Reconciliation Act of 1996. PMID- 19122868 TI - Applications of the Ugi reaction with ketones. AB - A convenient synthesis of highly functionalized, alpha,alpha-disubstituted amino acid amide derivatives has been accomplished by using cyclic and acyclic ketones as the carbonyl inputs in the Ugi multicomponent reaction. An application of this extension of the Ugi reaction to the synthesis of alpha,alpha-divinyl amino acids that may be cyclized via ring-closing metathesis to provide highly substituted pyrrolidines is described. PMID- 19122865 TI - Sleep and Its Disorders in Seniors. PMID- 19122869 TI - The Pauson-Khand Reaction as a New Entry to the Synthesis of Bridged Bicyclic Heterocycles: Application to the Enantioselective Total Synthesis of (-) Alstonerine. AB - The first application of the Pauson-Khand reaction (PKR) to the synthesis of azabridged bicyclic structures is described. Compounds containing azabicyclo[3.3.1]nonane and azabicyclo[3.2.1]octane rings fused to cyclopentenones were efficiently constructed via the PKR of cis-2,6-disubstituted N-acyl piperidine enyne substrates, many of which can be readily prepared from 4 methoxypyridine in a few steps. Moreover, the PKR of cis-2,6-disubstituted piperazine enynes allowed the preparation of diazabicyclo[3.3.1]nonanes fused to cyclopentenones. This new strategy for the synthesis of azabridged bicyclic frameworks was exploited as a key step in a concise, enantioselective total synthesis of the macroline alklaoid (-)-alstonerine. PMID- 19122870 TI - Directed Hydrozirconation of Homopropargylic Alcohols. AB - Homopropargylic alcohols undergo directed hydrozirconation with Schwartz reagent (Cp(2)ZrHCl) to generate vinyl-metal species in which the metal fragment is proximal to the alkoxide. Electrophilic trapping yields tri-substituted olefins in good yields with good control of regio- and stereochemistry. Experiments with a homopropargylic ether confirmed the role of the hydroxyl group in the directed hydrometalation. PMID- 19122871 TI - Getting the Word Out: Teaching Middle-School Children about Cardiovascular Disease. AB - Cardiovascular disease (CVD) has roots in childhood; since CVD begins early, a clear strong case for early education focused on CVD primary prevention exists. Scientists are not traditionally involved in disseminating health knowledge into public education. Similarly, public school teachers typically do not have access to biomedical research that may increase their students' health science literacy. One way to bridge the 'cultural' gap between researchers and school teachers is to form science education partnerships. In order for such partnerships to be successful, teams of scientists and teachers must 'translate' biomedical research into plain language appropriate for students.In this article, we briefly review the need for improving health literacy, especially through school-based programs, and describe work with one model scientist/teacher partnership, the Teacher Enrichment Initiatives. Examples of cardiovascular research 'translated' into plain language lessons for middle school students are provided and practical considerations for researchers pursuing a science education partnership are delineated. PMID- 19122872 TI - Bi(OTf)(3)-Catalyzed 5-Exo-Trig Cyclization via Halide Activation. AB - Lewis acid activation of allyl halides utilizing Bi(OTf)(3) resulted in cationic cyclization of alkenes with high efficiency. While other Lewis acids could catalyzed this process with highly substituted alkenes, bismuth salts demonstrated unique reactivity in come cases. This suggested that bismuth triflate possesses intersting halophilic properties. PMID- 19122873 TI - Respiratory Distress Syndrome Degrades the Fine Structure of the Non-Nutritive Suck In Preterm Infants. AB - AIMS AND OBJECTIVES: Suck development is a challenging hurdle for preterm infants who endure an extensive oxygen history due to respiratory distress syndrome (RDS). The fine structure of the non-nutritive suck (NNS) was studied in preterm infants according to RDS severity. DESIGN AND METHODS: Recordings of NNS were completed cribside in the neonatal intensive care unit (NICU) in 55 preterm infants distributed among one healthy control group and two RDS infant groups. NNS pressure amplitude (cmH(2)0) and within-burst suck cycle period (ms) were the dependent measures extracted from digitized records of pacifier nipple compression pressure. RESULTS AND CONCLUSIONS: RDS preterm infants demonstrated significant differences in NNS suck pressure amplitude compared to healthy preterm infants. Periods of oxygen supplementation restrict orofacial movement and limit orosensory experiences necessary for suck development and neural maturation. RDS infants may be excellent candidates for patterned oral stimulation programs designed to advance the maturation of sucking skills. PMID- 19122875 TI - Surface instability of an elastic half space with material properties varying with depth. AB - If a body with a stiffer surface layer is loaded in compression, a surface wrinkling instability may be developed. A bifurcation analysis is presented for determining the critical load for the onset of wrinkling and the associated wavelength for materials in which the elastic modulus is an arbitrary function of depth. The analysis leads to an eigenvalue problem involving a pair of linear ordinary differential equations with variable coefficients which are discretized and solved using the finite element method.The method is validated by comparison with classical results for a uniform layer on a dissimilar substrate. Results are then given for materials with exponential and error-function gradation of elastic modulus and for a homogeneous body with thermoelastically-induced compressive stresses. PMID- 19122874 TI - Modeling cancer patient populations in mice: complex genetic and environmental factors. AB - Genetic differences among individuals contribute to differential susceptibility to cancer and, undoubtedly, to variable efficacy and toxicity of pharmacological based therapeutics. Many of the specific molecular processes involved in human tumorigenesis have been elucidated and accurately modeled in mice. However, the current models used for drug testing do not accurately predict how new treatments will fare in clinical trials. More sophisticated models that treat cancer as a complex disease present within heterogenous patient populations will provide better predictive power to identify patients that may benefit from specific therapies or that may develop potential drug-induced toxicities. PMID- 19122876 TI - Polyelectrolyte Multilayers Fabricated from 'Charge-Shifting' Anionic Polymers: A New Approach to Controlled Film Disruption and the Release of Cationic Agents from Surfaces. AB - We report an approach to the design of 'charge-shifting' anionic polymers that provides control over the disruption of ultrathin polyelectrolyte multilayers in aqueous media. We demonstrate that the addition of citraconic anhydride to poly(allylamine) yields an anionic, carboxylate-functionalized polymer (polymer 2) that can be converted readily back to cationic poly(allylamine) in acidic environments. The incorporation of polymer 2 into polyelectrolyte multilayers thus provides an approach to the fabrication of films that are stable at neutral pH but that erode over a period of several days in acidic media (e.g., pH ~5). Experiments using a structural analog of polymer 2 with carboxylate side chains that do not hydrolyze readily provided support for the view that the disruption of these films occurred as a result of polymer side chain hydrolysis and a resulting change in the net charge of the polymers. Because this approach is based upon the use of anionic polymers to induce film instability, it provides a platform for the design of multilayers that can be used to provide control over the release of cationic film components. As proof of concept, we demonstrated that ultrathin films ~100 nm thick fabricated using polymer 2 sustain the release of fluorescently labeled PAH for up to four days when incubated at pH 5.0. The synthetic approach used here is modular and tunable and can be used to introduce anionic 'charge-shifting' character to a broad range of other polyamines. With further development, this approach could expand significantly the range of different cationic agents (e.g., cationic proteins, peptides, polymers, nanoparticles, etc.) that can be released or delivered from surfaces using polyelectrolyte multilayers. PMID- 19122877 TI - Chemopreventive doses of methylselenocysteine alter circadian rhythm in rat mammary tissue. AB - It is known that organic forms of selenium inhibit chemically induced rat mammary carcinogenesis, although the molecular basis remains to be elucidated. To identify signaling pathways involved in carcinogenesis that are also modulated by methylselenocysteine, we compared the global gene expression profiles in mammary tissues from pubescent female rats maintained on a selenium-supplemented (3 ppm) diet with those on a standardized diet after N-nitroso-N-methylurea. Whereas the selenium-enriched diet altered the steady-state levels of genes involved in various cellular functions, the most dramatic effect was the coordinated changes in the expression of multiple genes that regulate circadian rhythm. Normal mammary tissue of rats fed a standardized diet showed little circadian oscillation relative to liver tissue. By contrast, mammary tissue of rats maintained on the selenium-enriched diet showed a progressive, time-dependent increase in the expression of circadian gene Per2 and circadian-regulated transcription factor DBP. Our results further showed that the expression of Per2 and DBP mRNAs was significantly decreased in mammary tumors arising in rats on the selenium-enriched diet, but not in tumors of rats on the control diet, suggesting that selenium-induced elevation in the expression of circadian genes was incompatible with mammary carcinogenesis. Given the previously reported role of Per2 as a tumor suppressor, these observations suggest that Per2 is an important target of methylselenocysteine during chemoprevention in N-nitroso-N methylurea-induced rat mammary carcinogenesis, and for the first time provide a link between chemoprevention and circadian rhythm. PMID- 19122878 TI - Blood viscosity modulates tissue perfusion: sometimes and somewhere. AB - Each organ possesses specific properties for controlling microvascular perfusion. Such specificity provides an opportunity to design transfusion fluids that target thrombo-embolic or vasospasm-induced ischemia in a particular organ or that optimize overall perfusion from systemic shock. The role of viscosity in the design of these fluids might be underestimated, because viscosity is rarely monitored or considered in critical care decisions. Studies linking viscosity dependent changes of microvascular perfusion to outcome-relevant data suggest that whole blood viscosity is negligible as a determinant of microvascular perfusion under physiological conditions when autoregulation is effective. Because autoregulation is driven to maintain oxygen supply constant, the organism will compensate for changes in blood viscosity to sustain oxygen delivery. In contrast, under pathological conditions in the brain and elsewhere, increases of overall viscosity should be avoided - including all the situations where vascular autoregulatory mechanisms are inoperative due to ischemia, structural damage or physiologic dysfunction. As latter conditions are not to identify with high certainty, the risks that accompany therapeutic correction of blood viscosity are outweighing the benefits. The ability to bedside monitor blood viscosity and to link changes in viscosity to outcome parameters in various clinical conditions would provide more solid foundation for evidence-based clinical management. PMID- 19122879 TI - Slow and stepped re-warming after acute low temperature exposure do not improve survival of Drosophila melanogaster larvae. AB - We tested that hypothesis that slow re-warming rates would improve the ability of Drosophila melanogaster Meigen larvae to survive acute low temperature exposure. Four larval stages (1(st), 2(nd), 3(rd) instars and wandering stage 3(rd) instars) of four wild-type strains were exposed to -7 degrees C for periods of time expected to result in 90 % mortality. Larvae were then either directly transferred to their rearing temperature (21 degrees C), or returned to this temperature in a stepwise fashion (pausing at 0 and 15 degrees C) or by slow warming at 1 or 0.1 degrees C/min. We observed a reduced rapid cold-hardening effect and no general increase in survival of acute chilling in larvae re-warmed in a stepwise or slow fashion, and hypothesise that slow re-warming may result in accumulation of further chill injuries. PMID- 19122880 TI - The Use of Semantic Differential Scaling to Define the Multi-Dimensional Representation of Odors. AB - The mental representation elicited by smelling an odor often consists of multiple sensory and affective dimensions, yet, the richness of this elaboration is difficult to capture using methods to rate the intensity of these factors in isolation. Attempts to use language descriptors for olfactory experience have also been shown to be rather limited; among non-specialists, there is no universally accepted system for describing odors, leading to greater reliance on specific item associations. In this study we explored the utility of semantic differential scaling for illustrating the various dimensions of olfactory experience. 300 volunteers rated thirty distinct odorants using 50 SDS adjectives. Three factors emerged from the analysis (based on 17 adjective-pairs) accounting for 53% of the variance, and corresponding to the evaluation, potency and activity dimensions identified for other stimulus types. SD scaling appears to be a viable method for identifying the multiple dimensions of mental representation evoked when smelling an odorant and may prove a useful tool for both consumer and basic research alike. PRACTICAL APPLICATIONS: Although numerous methods of classifying odors have been developed, little agreement has been achieved on the dimensions that are useful to both basic and consumer research. The identification of a set of Semantic Differential adjectives which are relevant to olfactory experience can become a useful tool for classifying the qualitative and affective basis on which odorants differ.. In particular, the degree to which odorants evokes multi-dimensional representations from other sensory modalities (visual, auditory, somatosensory or gustatory), can be usefully applied in the arena of product development both within and across cultures. PMID- 19122881 TI - Effects of Small-Group Tutoring with and without Validated Classroom Instruction on At-Risk Students' Math Problem Solving: Are Two Tiers of Prevention Better Than One? AB - The purpose of this study was to assess the effects of small-group tutoring with and without validated classroom instruction on at-risk (AR) students' math problem solving. Stratifying within schools, 119 3(rd)-grade classes were randomly assigned to conventional or validated problem-solving instruction (Hot Math [schema-broadening instruction]). Students identified as AR (n = 243) were randomly assigned, within classroom conditions, to receive Hot Math tutoring or not. Students were tested on problem-solving and math applications measures before and after 16 weeks of intervention. Analyses of variance, which accounted for the nested structure of the data, revealed the tutored students who received validated classroom instruction achieved better than tutored students who received conventional classroom instruction (ES = 1.34). However, the advantage for tutoring over no tutoring was similar whether or not students received validated or conventional classroom instruction (ESs = 1.18 and 1.13). Tutoring, not validated classroom instruction reduced the prevalence of math difficulty. Implications for responsiveness-to-intervention prevention models and for enhancing math problem-solving instruction are discussed. PMID- 19122882 TI - The Impact of Salmon Bias on the Hispanic Mortality Advantage: New Evidence from Social Security Data. AB - A great deal of research has focused on factors that may contribute to the Hispanic mortality paradox in the United States. In this paper, we examine the role of the salmon bias hypothesis - the selective return of less-healthy Hispanics to their country of birth - on mortality at ages 65 and above. These analyses are based on data drawn from the Master Beneficiary Record and NUMIDENT data files of the Social Security Administration. These data provide the first direct evidence regarding the effect of salmon bias on the Hispanic mortality advantage. Although we confirm the existence of salmon bias, it is of too small a magnitude to be a primary explanation for the lower mortality of Hispanic than NH white primary social security beneficiaries. Longitudinal surveys that follow individuals in and out of the United States are needed to further explore the role of migration in the health and mortality of foreign-born US residents and factors that contribute to the Hispanic mortality paradox. PMID- 19122883 TI - Stages of the Demographic Transition from a Child's Perspective: Family Size, Cohort Size, and Children's Resources. PMID- 19122884 TI - Irregular and Semi-Regular Polyhedral Models for Rous Sarcoma Virus Cores. AB - Whereas many viruses have capsids of uniquely defined sizes that observe icosahedral symmetry, retrovirus capsids are highly polymorphic. Nevertheless, they may also be described as polyhedral foldings of a fullerene lattice on which the capsid protein (CA) is arrayed. Lacking the high order of symmetry that facilitates the reconstruction of icosahedral capsids from cryo-electron micrographs, the three-dimensional structures of individual retrovirus capsids may be determined by cryo-electron tomography, albeit at lower resolution. Here we describe computational and graphical methods to construct polyhedral models that match in size and shape, capsids of Rous sarcoma virus (RSV) observed within intact virions [8]. The capsids fall into several shape classes, including tubes, "lozenges", and "coffins". The extent to which a capsid departs from icosahedral symmetry reflects the irregularity of the distribution of pentamers, which are always 12 in number for a closed polyhedral capsid. The number of geometrically distinct polyhedra grows rapidly with increasing quotas of hexamers, and ranks in the millions for RSV capsids, which typically have 150 - 300 hexamers. Unlike the capsid proteins of icosahedral viruses that assume a minimal number of quasi equivalent conformations equal to the triangulation number (T), retroviral CAs exhibit a near-continuum of quasi-equivalent conformations - a property that may be attributed to the flexible hinge linking the N- and C-terminal domains. PMID- 19122885 TI - Diagnosis versus dialogue: oral testimony and the study of pediatric pain. AB - Through the perspectives of the children, this essay examines the communication between pediatric pain patients and their doctors. Based upon the oral history responses of thirty-two patients with chronic pain present for evaluation at the Pediatric Pain Clinic at UCLA, oral testimony was employed to uncover a wide range of topics related to a child's experience with pain such as family dynamics, how and when pain became a life-changing factor, coping strategies, and external sources that contribute to the child's understanding of pain. Most important, children were encouraged to explain what it was like to be in pain, not only to describe symptoms but also to share their dreams and hopes, their fears and uncertainties -- as well as the place of pain in their world. PMID- 19122886 TI - The Long-Term Consequences of Childbearing: Physical and Psychological Well-Being of Mothers in Later Life. AB - Growing evidence points to relationships between patterns of childbearing and health outcomes for mothers; yet a need remains to clarify these relationships over the long-term and to understand the underlying mechanisms. Using data from the National Longitudinal Survey of Mature Women (N=1,608), I find that the long term consequences of childbearing vary by health outcome. Early childbearing is associated with higher risk of ADL limitations at ages 65-83, though effects appear stronger among white than black mothers until SES is controlled. Early childbearing is also associated with greater levels of depressive symptomatology, though this association is mediated by SES and health. Late childbearing is associated with more depressive symptoms net of early life and current SES, child proximity and support, and physical health. Finally, I find no significant effects of high parity. These findings emphasize the need to better understand the mechanisms linking childbearing histories to later physical and psychological well-being. PMID- 19122887 TI - Heavy Drinking and Polydrug Use among College Students. AB - Excessive alcohol consumption is a serious problem on college campuses but may not be adequately captured by traditional methods of defining binge drinking. This study examined a new approach to categorizing alcohol use and its relationship with illicit drug use. A survey was administered to 484 college students ages 18 to 25. Drinkers were divided into three groups based on the number of typical drinks consumed per day: "light"-1 to 4 (n=182); "moderate"-5 to 9 (n=173); and "heavy"-10+ (n=56). Heavy drinkers could be differentiated from moderate and light drinkers on age of onset of alcohol use, illicit drug use, and frequency of illicit drug use. A binary categorization of "binge" vs. "nonbinge" drinking may obscure important differences within binge drinkers. These findings have implications for prevention, as well as clinical risk assessment of college student drinkers for adverse consequences of concomitant alcohol and illicit drug consumption. PMID- 19122888 TI - Response to early literacy instruction in the United States, Australia, and Scandinavia: A behavioral-genetic analysis. AB - Genetic and environmental influences on early reading and spelling at the end of kindergarten and Grade 1 were compared across three twin samples tested in the United States, Australia, and Scandinavia. Proportions of variance due to genetic influences on kindergarten reading were estimated at .84 in Australia, .68 in the U.S., and .33 in Scandinavia. The effects of shared environment on kindergarten reading were estimated at .09 in Australia, .25 in the U.S., and .52 in Scandinavia. A similar pattern of genetic and environmental influences was obtained for kindergarten spelling. One year later when twins in all three samples had received formal literacy instruction for at least one full school year, heritability was similarly high across country, with estimated genetic influences varying between .79 and .83 for reading and between .62 and .79 for spelling. These findings indicate that the pattern of genetic and environmental influences on early reading and spelling development varies according to educational context, with genetic influence increasing as a function of increasing intensity of early instruction. Longitudinal analyses revealed genetic continuity for both reading and spelling between kindergarten and Grade 1 across country. However, a new genetic factor comes into play accounting for independent variance in reading at Grade 1 in the U.S. and Scandinavia, suggesting a change in genetic influences on reading. Implications for response-to-instruction are discussed. PMID- 19122889 TI - Synthetic Studies of 3-(3-Fluorooxindol-3-yl)-l-alanine. AB - Oxidative fluorination of several protected tryptophans 8b-g with Selectfluor proceeded smoothly in aqueous media to give a diastereomeric mixture of the corresponding 3-fluorooxindoles 9b-g. Attempted deprotection of the 3 fluorooxindoles 9b-g under various conditions did not afford 3-(3-fluorooxindol-3 yl)-l-alanine (6). Reaction of the suitably protected tryptophan derivative 16 with Selectfluor produced the fluorinated product 17. Simultaneous cleavage of all protective groups of 17 under acidic conditions successfully gave the target compound 6 in excellent yield. PMID- 19122890 TI - A Note on Conditional AIC for Linear Mixed-Effects Models. AB - The conventional model selection criterion AIC has been applied to choose candidate models in mixed-effects models by the consideration of marginal likelihood. Vaida and Blanchard (2005) demonstrated that such a marginal AIC and its small sample correction are inappropriate when the research focus is on clusters. Correspondingly, these authors suggested to use conditional AIC. The conditional AIC is derived under the assumptions of the variance-covariance matrix or scaled variance-covariance matrix of random effects being known. We develop a general conditional AIC but without these strong assumptions. This allows Vaida and Blanchard's conditional AIC to be applied in a wide range. Simulation studies show that the proposed method is promising. PMID- 19122892 TI - "The Heart Truth:" Using the Power of Branding and Social Marketing to Increase Awareness of Heart Disease in Women. AB - In September 2002, the National Heart, Lung, and Blood Institute launched The Heart Truth, the first federally-sponsored national campaign aimed at increasing awareness among women about their risk of heart disease. A traditional social marketing approach, including an extensive formative research phase, was used to plan, implement, and evaluate the campaign. With the creation of the Red Dress as the national symbol for women and heart disease awareness, the campaign integrated a branding strategy into its social marketing framework. The aim was to develop and promote a women's heart disease brand that would create a strong emotional connection with women. The Red Dress brand has had a powerful appeal to a wide diversity of women and has given momentum to the campaign's three-part implementation strategy of partnership development, media relations, and community action. In addition to generating its own substantial programming, The Heart Truth became a catalyst for a host of other national and local educational initiatives, both large and small. By the campaign's fifth anniversary, surveys showed that women were increasingly aware of heart disease as their leading cause of death and that the rise in awareness was associated with increased action to reduce heart disease risk. PMID- 19122891 TI - Prediction of Cis-Regulatory Elements Controlling Genes Differentially Expressed by Retinal and Choroidal Vascular Endothelial Cells. AB - Cultured endothelial cells of the human retina and choroid demonstrate distinct patterns of gene expression. We hypothesized that differential gene expression reflected differences in the interactions of transcription factors and respective cis-regulatory motifs(s) in these two emdothelial cell subpopulations, recognizing that motifs often exist as modules. We tested this hypothesis in silico by using TRANSFAC Professional and CisModule to identify cis-regulatory motifs and modules in genes that were differentially expressed by human retinal versus choroidal endothelial cells, as identified by analysis of a microarray data set. Motifs corresponding to eight transcription factors were significantly (p < 0.05) differentially abundant in genes that were relatively highly expressed in retinal (i.e., GCCR, HMGIY, HSF1, p53, VDR) or choroidal (i.e., E2F, YY1, ZF5) endothelial cells. Predicted cis-regulatory modules were quite different for these two groups of genes. Our findings raise the possibility of exploiting specific cis-regulatory motifs to target therapy at the ocular endothelial cells subtypes responsible for neovascular age-related macular degeneration or proliferative diabetic retinopathy. PMID- 19122894 TI - Mother-Child Attachment and Social Anxiety Symptoms in Middle Childhood. AB - Literature suggests that parent-child attachment and anxiety symptoms are related. One purpose of the present study was to assess whether attachment patterns relate differentially to social anxiety aspects (fear of negative evaluation, social anxiety and distress in new situations, and generalized anxiety and distress). The second purpose was to investigate these links both longitudinally and concurrently in middle childhood. Children in grades 3 and 5 (N = 74) completed measures of secure, ambivalent, and avoidant attachments with mothers and a measure of social anxiety symptoms in grade 5. Longitudinal analyses showed that ambivalent attachment was most consistently related to social anxiety. Concurrent measures of attachment and social anxiety showed that lower attachment security and higher ambivalent attachment were most consistently related to higher social anxiety. Concurrent attachment predicted variance in social anxiety after controlling for earlier attachment. Findings suggest that anxiety interventions might target attachment. PMID- 19122893 TI - Actions speak louder than words: An elaborated theoretical model of the social functions of self-injury and other harmful behaviors. AB - The question of why some people do things that are intentionally harmful to themselves continues to puzzle scientists, clinicians, and the public. Prior studies have demonstrated that one fairly extreme, direct form of self-harm, non suicidal self-injury (NSSI), is maintained by both automatic (i.e., intrapersonal) as well as social (i.e., interpersonal) reinforcement. However, the majority of theoretical and empirical papers on this topic focus almost exclusively on the automatic functions. The purpose of this paper is to provide a more comprehensive analysis of the social functions of NSSI. Evidence is presented supporting the notion that NSSI is maintained by social reinforcement in at least a substantial minority of instances. Moreover, an elaborated theoretical model of the social functions of NSSI is outlined that proposes that this behavior represents a high intensity social signal used when less intense communication strategies fail (e.g., speaking, yelling, crying). The model further proposes that NSSI can serve not only as a signal of distress that is reinforced primarily by the caregiving behavior it elicits from others, but that it also can serve as a signal of strength and fitness that is reinforced by warding off potential threats (e.g., peer victimization), and in some cases can strengthen affiliation with others. Support for this theoretical model is drawn from diverse literatures including psychology, evolutionary biology, and cultural anthropology. The paper concludes with specific recommendations for empirical tests of the proposed model of the social functions of NSSI, as well as other harmful behaviors such as alcohol and drug use. PMID- 19122895 TI - DNA-binding drugs caught in action: the latest 3D pictures of drug-DNA complexes. AB - In this paper, we review recent DNA-binding agents that are expected to influence the field of DNA-targeting. We restrict ourselves to binders for which the three dimensional structure in complex with DNA or RNA has been determined by X-ray crystallography or NMR. Furthermore, we primarily focus on unprecedented ways of targeting peculiar DNA structures, such as junctions, quadruplexes, and duplex DNAs different from the B-form. Classical binding modes of small molecular weight compounds to DNA, i.e. groove binding, intercalation and covalent addition are discussed in those cases where the structures represent a novelty. In addition, we review 3D structures of triple-stranded DNA, of the so-called Peptide Nucleic Acids (PNAs), which are oligonucleotide bases linked by a polypeptide backbone, and of aptamers, which are DNA or RNA receptors that are designed combinatorially. A discussion on perspectives in the field of DNA-targeting and on sequence recognition is also provided. PMID- 19122897 TI - [NH4]12[(MoO2)2O(HPO4)2]4[PO4]X, X = Cl, Br--new porous layer molybdenophosphates with embedded multiple anionic and cationic guests. AB - The compounds [NH4]12[(MoO2)2O(HPO4)2]4[PO4]X, X = Cl, Br are constructed from molybdenophosphate layers of unique topology that are penetrated by channels containing perfectly ordered, alternating ammonium cations and free orthophosphate (PO4(3-)) or halide (Cl-, Br-) anions. PMID- 19122896 TI - A unique chair-shaped hexanuclear Cu(I) metallamacrocyclic C2H4 adduct encapsulating a BF4- anion. AB - Three novel Cu(I)-C2H4 adducts bearing 4-(2-pyridyl)pyrimidine were self assembled, and the roles of anion and solvent were proved in the formation process. PMID- 19122898 TI - Twice silicon-induced C-H activation and tautomerisation of a beta-diketiminato ligand and formation of new types of N-heterocyclic silanes. AB - Unexpected formation of the novel dianionic N,C-chelate ligand LD2- [HC[C(Me) NR][C(NHR)=CH]]2- occurs by a twice silicon-assisted C-H bond activation of a terminal methyl group on a beta-diketiminato ligand backbone, starting from the corresponding lithium beta-diketiminide and silicon tetrabromide to give the new N-heterocyclic dibromosilanes and . PMID- 19122900 TI - Functionalised azetidines as ligands: pyridyl-complemented coordination. AB - A relatively simple azetidine derivative with 2-aminoethyl and 2-pyridyl substituents functions as a tridentate ligand towards Cu(II), giving species in which the CuN3 unit is essentially planar and that, in the solid state, show a subtle balance between mononuclear and binuclear forms, allowing both to be simultaneously present in some lattices. Displacement of the ligand from Cu(II) by the use of cyanide results in the formation of cyanogen, which appears to slowly react with the free amine to give an oxamidine characterised in its doubly deprotonated form as its binuclear Ni(II) complex. PMID- 19122899 TI - NMR spectroscopy and molecular modelling studies of nitrosylcobalamin: further evidence that the deprotonated, base-off form is important for nitrosylcobalamin in solution. AB - The structure of nitrosylcobalamin (NOCbl) in solution has been studied by NMR spectroscopy and the 1H and 13C NMR spectra have been assigned. 13C and 31P NMR chemical shifts, the UV-vis spectrum of NOCbl and the observed pKbase-off value of approximately 5.1 for NOCbl provide evidence that a significant fraction of NOCbl is present in the base-off, 5,6-dimethylbenzimidazole (DMB) deprotonated, form in solution. NOE-restrained molecular mechanics modelling of base-on NOCbl gave annealed structures with minor conformational differences in the flexible side chains and the nucleotide loop position compared with the X-ray structure. A molecular dynamics simulation at 300 K showed that DMB remains in close proximity to the alpha face of the corrin in the base-off form of NOCbl. Simulated annealing calculations produced two major conformations of base-off NOCbl. In the first, the DMB is perpendicular to the corrin and its B3 nitrogen is about 3.1 A away from and pointing directly at the metal ion; in the second the DMB is parallel to and tucked beneath the D ring of the corrin. PMID- 19122901 TI - Functionalised azetidines as ligands: species derived by selective alkylation at substituent-nitrogen. AB - Selective functionalisation of the tridentate ligand 1-(2-aminoethyl)-3-methyl-3 (2-pyridyl)azetidine at its terminal amino-nitrogen atom can be readily achieved by both reductive alkylation and simple alkylation reactions to give tri-, quadri , quinque- and sexi-dentate derivatives. Simple alkylation by 2-picolinyl chloride provides the only example of a second reaction pathway where the azetidine ring of the reactant has undergone activation towards ring opening. Structural characterisation of the Cu(II) complexes of these ligands has revealed several remarkable aspects of their solid-state coordination chemistry, including the formation of infinite helical aggregates through pi-stacking and tetramerisation through carboxylate bridging, as well as further examples of the crystallisation of mixed species found to be rather common with complexes of the parent ligand. PMID- 19122902 TI - Noble metal ionic catalysts: correlation of increase in CO oxidation activity with increasing effective charge on Pd ion in Pd ion substituted Ce1-xMxO2-delta (M = Ti, Zr and Hf). AB - Pd ion substituted Ce1-xMxO2-delta (M = Ti, Zr, Hf) have been prepared by a single step solution combustion method. Two atom% Pd ion substitution in the title compounds is confirmed by X-ray diffraction (XRD) and Pd ion charge state and redox properties have been determined by X-ray photoelectron spectroscopy (XPS) and H2/TPR studies. While Pd ion in CeO2 (Ce0.98Pd0.02O2-delta) showed higher catalytic activity for CO oxidation than Pd metal impregnated over CeO2, further increase in the catalytic activity is observed with Pd ion in Ce1 xTi(Hf)xO2 and a decrease in the activity with Pd ion in Ce1-xZrxO2. Effective charge on Pd ion could be varied by its substitution in these solid solutions Ce1 xMxO2 (M = Ti, Zr & Hf) and also in TiO2 compared to Pd ion in PdO. Effective positive charge on Pd ion is determined from the core level binding energy shift of Pd(3d5/2) peak with respect to Pd metal. Rate of CO oxidation increased and activation energy decreased with increase in effective charge on the Pd ion in the Pd ion substituted fluorites. PMID- 19122903 TI - Dinuclear fluoro-peroxovanadium(v) complexes with symmetric and asymmetric peroxo bridges: syntheses, structures and DFT studies. AB - Two new dinuclear fluoro peroxovanadium(v) complexes, Cs3[V2O2(O2)4F] x H2O (1) and Cs3[V2O2(O2)3F3] x 2HF x H2O (2), were prepared and characterized by elemental analysis, IR spectroscopy, thermal analysis and X-ray crystallography. While the anion in possesses an asymmetric structure with a micro-eta1:eta2 bridging peroxo group, the [V2O2(O2)3F3]3- ion in exhibits a symmetrical structure with a unique mu-fluoro and micro-eta2:eta2 peroxo double bridge. The X ray structure data were compared with equilibrium and vibrationally-averaged (effective) DFT calculated geometries. The decomposition reactions of and in aqueous solution were studied by 51V NMR spectroscopy. The calculations of vibrationally averaged NMR chemical shifts (DFT-GIAO) were used to support the empirical assignment of NMR signals and afforded excellent agreement with experimental values for the studied peroxovanadium species. The ESI mass spectra of the prepared compounds are in accordance with the assignment of NMR spectra and with DFT study. PMID- 19122904 TI - Dendritic (phosphine)gold(I) thiolate complexes: assessment of the molecular size through PGSE NMR studies. AB - The reactions of the tetraphosphine ligand DAB-G0-(PPh2)4 (DAB = 1,4 diaminobutane; G0 = Generation 0) or the octaphosphine ligand DAB-G1-(PPh2)8 (G1 = Generation 1) with the gold precursor [AuCl(tht)] (tht =tetrahydrothiophene) and the corresponding 4-substituted benzenethiolate lead to the (phosphine)gold(I) thiolate complexes [Au4(S-C6H4-X)4[(DAB-G0-(PPh2)4]] (X = F (11), MeO (12), Me (13) and NO2 (14)) and [Au8(S-C6H4-X)8[(DAB-G1-(PPh2)4]] (X = F (15), MeO (16), Me (17) and NO2 (18)). Complexes [Au4Cl4[(DAB-G0-(PPh2)4]] and [Au4(S-C6H4-OMe)4[(DAB-G0-(PPh2)4]] have been characterized by X-ray diffraction studies showing tetranuclear gold complexes in which the P-Au-X (X = Cl or S) structural units do not display aurophilic interactions. PGSE NMR studies of free ligands EN-G0-(PPh2)(4) (EN = 1,2-ethylenediamine), DAB-G0-(PPh2)(4), DAB-G1 (PPh2)8 and the (phosphine)gold(I) thiolate complexes permit the evaluation and comparison of the different molecular sizes depending on the ligand and the dendrimer generation. PMID- 19122905 TI - Growth of La1-xSrxFeO3 thin films by atomic layer deposition. AB - Thin films of La1-xSrxFeO3 have been prepared by the ALD (atomic layer deposition) technique using La(thd)3 (Hthd = 2,2,6,6-tetramethylheptane-3,5 dione), Sr(thd)2, Fe(thd)3, and ozone as precursors. A so-called ALD window is found in the temperature range 200 to 360 degrees C for LaFeO3. The effect of the pulsing procedure for the precursors on the composition of the films is examined. The results are discussed in relation to a model which ascribes differences between pulsed and obtained stoichiometries to individually different surface area demands of the precursors. The La1-xSrxFeO3 films turned out to contain only small amounts of carbonate impurities despite the fact that films prepared from Sr(thd)2 and ozone under the same conditions contains virtually pure SrCO3. Films of La1-xSrxFeO3 have been deposited on substrates of (amorphous) soda-lime glass and single crystals of Si(100), SrTiO3(100), and LaAlO3(012). Annealed films on soda-lime glass and Si(100) substrates turned out to be polycrystalline with virtually random orientation of the crystallites. Those on MgO(100) and SrTiO3(100) substrates showed some degree of crystal orientation, whereas the annealed films on LaAlO3(012) proved to contain distinctly oriented crystallites with mosaic features. PMID- 19122906 TI - P,N-Containing cyclophanes with large helical hydrophobic cavities: prospective precursors for the design of a molecular reactor. AB - The highly effective self-assembly process between 2,4,6 tris(isopropyl)phenylphosphine or mesitylphosphine, formaldehyde and 1,4 bis(alpha-(4'-aminophenyl)isopropyl)benzene results in the formation of a novel type of heterocyclophane which is able to encapsulate organic molecules as well as being a potential ligand for transition metals inside the intramolecular cavity. Compounds of this type can be regarded as potential precursors for a new kind of molecular reactors. PMID- 19122907 TI - Peculiar electronic and vibrational properties of metal-dithiolenes (Ni, Au) based on 1,2,5-thiadiazole-3,4-dithiolato. AB - In this paper, we present a theoretical study based on DFT methods using functionals implemented in Gaussian03, to obtain geometry optimizations, harmonic frequencies, IR intensities and Raman scattering activities of the triad [Ni(tdas)2]z [z = 0 (); z = 1- (); z = 2- ()] and of [Au(tdas)2]- (3) with the aim of elucidating the nature of the bonding in these complexes and checking whether C-C stretching vibrations are suitable spectroscopic markers to assign the 'innocent-non-innocent' character of the ligand. Geometry optimization at the density functional theory level was performed, and geometrical parameters were obtained in good agreement with the experimental structural data for the Ni complexes. Following the geometrical changes upon reduction, the Ni-S, N-S and C S distances undergo a small increase, while the C-C length does not change significantly. Thus, unlike what is generally found for non-benzenoid dithiolenes, no shift of the nu(C-C) vibration is expected on reduction as observed for 1 and 2. The vibrational spectra are very accurately reproduced by the calculations. The less satisfactory agreement obtained for the gold complex is probably due to the deviation of the structurally characterized sample from the D2h symmetry obtained in the calculations, and/or overestimation of the Au-S distances, typical of the DFT functionals used. Calculations indicated that the electroactive orbital, HOMO in and , SOMO in and LUMO in , is a highly delocalized pi-orbital with a predominant S-ligand character, and that the total spin density in is delocalized over the ligands and the nickel atom, in agreement with EPR data. From the obtained results we can assign a non-innocent character to the tdas ligand and show that the C-C stretching vibrations are not suitable spectroscopic markers to reflect this feature. PMID- 19122909 TI - Cadmium dicarboxylate coordination polymers incorporating a long-spanning organodiimine: in situ alkene isomerization and an unprecedented chiral 6(5)8 two dimensional network. AB - Reaction of cadmium nitrate, maleic acid, and N,N'-bis(4-pyridylmethyl)piperazine (bpmp) in a methanol/water mixed solvent system at 25 degrees C resulted in an unexpected in situ isomerization and formation of [[Cd(fumarate)(bpmp)(H2O)2] x 2H2O]n (1), which exhibits a system of mutually interpenetrated [Cd(fumarate)(bpmp)(H(2)O)]n grid-like coordination polymer layers. A similar reaction using cadmium chloride and succinic acid produced [[Cd(succinate)(bpmp)(CH3OH)] x 2H2O]n (2), which manifests a chiral two dimensional coordination polymer network with an unprecedented 6(5)8 topology. The differences in the conformations of the four-carbon dicarboxylate tethers (anti in , gauche in ) and cadmium coordination environment (octahedral in , pentagonal bipyramidal in ) play a significant role in structure direction during self-assembly. Thermal properties of and are also reported. PMID- 19122908 TI - Does the metal influence non-covalent binding of complexes to DNA? AB - Electrospray ionisation mass spectrometry, absorption spectrophotometry and circular dichroism spectroscopy were used to investigate the binding of a series of nickel complexes with the general formula [Ni(phen)2L]2+ (L = phen, dpq, dpqC and dppz) to a double stranded DNA hexadecamer. In addition, the binding of the complexes to pUC9 negatively supercoiled plasmid DNA was examined using gel electrophoresis, and their ability to inhibit DNA transcription was measured. Each of the above techniques showed that DNA binding strengthened as the size of the unique ligand L was increased. Comparison of the above results with those obtained previously, and presented here for the first time for the analogous series of ruthenium complexes [Ru(phen)2L]2+, showed that changing the metal ion from nickel to ruthenium consistently resulted in significant increases in DNA binding affinity. PMID- 19122910 TI - Theoretical investigation on the dimerization of the deprotonated aquo ion of Al(III) in water. AB - Reaction pathways, solvent effects and energy barriers have been investigated for the dimerization of the deprotonated aquo ion of Al(III) in aqueous solution by performing supramolecule density functional theory calculations. Two competing reaction pathways were investigated, sharing a common first step and third step, i.e. the formation of the aggregate II of two aluminium monomers and the doubly bridged dimer. One pathway involves a nucleophilic attack to undercoordinated metal center in the first step and then the loss of a coordinated water molecule. Another pathway involves a water exchange reaction in the first step and then the formation of the hydroxo bridge. The calculated results indicate that both pathways I and II are possible in aqueous solution. The direct participation of the solvent water molecule facilitates the dimerization, but the extremely large solvent shifts of the energy barriers for each reaction are attributed mainly to the bulk effect. The computed activation energies for the water exchange reactions are in good agreement with the available experimental values, namely, the calculated value 37.5 kJ mol(-1) compared to the experimental value 36.4 (+/ 5) kJ mol(-1). In agreement with experimental observations in aqueous solution, the calculated results favor the transformation of singly-bridged to doubly bridged aluminium ion, which is helpful to understand the complicated hydrolytic polymerizaiton of Al(III). PMID- 19122911 TI - The d3/d2 alkyne complexes [MX2(eta-RC[triple bond, length as m-dash]CR)Tp']z (X = halide, z = 0 and 1+): final links in a d6-d2 redox family tree. AB - The d4 halide complexes [MX(CO)(eta-RC[triple bond, length as m-dash]CR)Tp'] [R = Me, M = W, X = F; R = Ph, M = Mo or W, X = F or Cl; Tp' = hydrotris(3,5 dimethylpyrazolyl)borate] undergo two-electron oxidation in the presence of a halide source to give the d2 monocations [MX1X2(eta-PhC[triple bond, length as m dash]CPh)Tp']+ (R = Me, M = W, X1 = X2 = F; R = Ph, M = Mo, X1 = X2 = F or Cl; M = W, X1 = X2 = F or Cl; X1 = F, X2 = Cl). Each monocation (R = Ph) shows two reversible one-electron reductions (the second process was not detected for R = Me) corresponding to the stepwise formation of the neutral d3 and monoanionic d4 analogues, [MX1X2(eta-PhC[triple bond, length as m-dash]CPh)Tp'] and [MX1X2(eta PhC[triple bond, length as m-dash]CPh)Tp']- respectively; the potentials for the two processes can be 'tuned' over a range of ca. 1.0 V by varying M and X. Chemical one-electron reduction of [MX2(eta-PhC[triple bond, length as m dash]CPh)Tp']+ gave [MX2(eta-PhC[triple bond, length as m-dash]CPh)Tp'] (M = Mo or W, X = F or Cl). X-Ray structural studies on the redox pairs [WX2(eta PhC[triple bond, length as m-dash]CPh)Tp']z (X = F and Cl, z = 0 and 1+) show the alkyne to bisect the X-W-X angle in the d2 cations but align more closely with one M-X bond in the neutral d3 molecules, consistent with the anisotropic ESR spectra of the latter; the solution ESR spectrum of [MoF2(eta-PhC[triple bond, length as m-dash]CPh)Tp'] showed equivalent fluorine atoms, i.e the alkyne oscillates at room temperature. The successful isolation of [MX2(eta-PhC[triple bond, length as m-dash]CPh)Tp']+ and [MX2(eta-PhC[triple bond, length as m dash]CPh)Tp'] completes a series in which d6 to d2 alkyne complexes are linked in a redox family tree by sequential one-electron transfer and substitution reactions. The implications for such trees in the production of new species and the selective synthesis of paramagnetic complexes acting as synthetically useful 'alkyne radicals' are discussed. PMID- 19122912 TI - Reactivity of the zwitterionic ligand EtNHC(S)Ph2P[double bond, length as m dash]NPPh2C(S)NEt towards [Ru3(CO)12]. Sulfur transfer and ligand fragmentation leading to the methideylamide [-N(Et)-CH(R)-] micro3-bridging moiety. AB - The reaction of EtNHC(S)Ph2P[double bond, length as m-dash]NP+Ph2C(S)N(-)Et (HEtSNS) with [Ru3(CO)12] has been carried out under two different experimental conditions: in the first case [Ru3(CO)12], previously turned into the labile intermediate [Ru3(CO)10(CH3CN)2], afforded, at room temperature in dichloromethane, the trinuclear clusters [Ru3(CO)11(CNEt)] (1), [Ru3(CO)9(micro H)[(micro-S:kappa-P)Ph2PN[double bond, length as m-dash]PPh2C(S)NEt]] (2), [Ru3(CO)9(micro-H)[(micro-S:kappa-P)Ph2PN[double bond, length as m-dash]P(S)Ph2]] (3) and [Ru3(CO)10[(micro-kappa2P)Ph2PNHPPh2]] (3). Ligand fragmentation occurs via loss of EtNC or EtNCS, without sulfur transfer to the cluster core. In the second case, [Ru3(CO)12] reacted with HEtSNS in toluene at 70 degrees C, giving the trinuclear clusters , [Ru3(CO)7(CNEt)(micro3-S)[(micro2-N:eta1-C:kappa1 P)Ph2PN[double bond, length as m-dash]PPh2C(H)NEt]] (6), [Ru3(CO)8)(micro3 S)[(micro2-N:eta-C:kappa-P)Ph2PN[double bond, length as m-dash]PPh2C(H)NEt]] (6) and [Ru3(CO)6(micro3-CO)(micro3-S)(EtNC)[(micro-kappa2P) Ph2PNHPPh2]] (7). The last three compounds derive from ligand fragmentation and sulfur transfer to the metal cluster. All compounds were characterized by spectroscopy (NMR, IR) and the molecular structures of , and were determined by single-crystal X-ray diffraction. Cluster preserves the original Ru3 triangular core in which an edge is bridged by a hydride ligand and by the sulfur atom of the Ph2PN[double bond, length as m-dash]PPh2C(S)NEt ligand. Cluster shows an open triangle of Ru atoms capped by a micro3-sulfide and by the unprecedented methideylamide -N(Et)CH(R) micro3-bridging moiety of the Ph2PN[double bond, length as m-dash]PPh2C(H)NEt ligand. It formally derives from cluster by substitution of ethyl isonitrile with one CO molecule. Finally, cluster displays a Ru3(micro3-S)(micro3-CO) trigonal bipyramidal core. PMID- 19122913 TI - Rare-earth metal complexes stabilized by amino-phosphine ligand. Reaction with mesityl azide and catalysis of the cycloaddition of organic azides and aromatic alkynes. AB - Stoichiometric reactions between mesityl azide (MesN3, Mes = 2,4,6-C6H2Me3) and amino-phosphine ligated rare-earth metal alkyl, LLn(CH2SiMe3)2(THF) (L = (2,6 C6H3Me2)NCH2C6H4P(C6H5)2; Ln = Lu (1a), Sc (1b)), amide, LLu(NH(2,6 C6H3iPr2))2(THF) (2) and acetylide at room temperature gave the amino-phosphazide ligated rare-earth metal bis(triazenyl) complexes, [L(MesN3)]Ln[(MesN3)(CH2SiMe3)]2 (Ln = Lu (3a); Sc (3b)), bis(amido) complex [L(MesN3)]Lu[NH(2,6-C6H3iPr2)]2 (4), and bis(alkynyl) complex (5) (L(MesN3)Lu (C[triple bond, length as m-dash]CPh)2)2, respectively. The triazenyl group in coordinates to the metal ion in a rare eta2-mode via N(beta) and N(gamma) atoms, generating a triangular metallocycle. The amino-phosphazide ligand, L(MesN3), in , and chelates to the metal ion in a eta3-mode via N(alpha) and N(gamma) atoms. In the presence of excess phenylacetylene, complex isomerized to , where the triazenyl group coordinates to the metal ion in a eta3 mode via N(alpha) and N(gamma) atoms. Complexes , , and have shown an unprecedented catalytic activity towards the cycloaddition of organic azides and aromatic alkynes to afford 1,5 disubstituted 1,2,3-triazoles selectively. PMID- 19122914 TI - Self-assembly supramolecular architectures of chromium(III) complexes using croconate as building block. AB - The synthesis, X-ray structure and magnetic properties of polynuclear Cr(III) complexes with the croconate ligand (C5O5(2-)) are reported: [Cr(mu-C5O5)(micro OH)(OH2)2]2 x 6H2O (, orthorombic, space group Cmca, a = 6.741(7) A, b = 18.841(9) A, c = 16.886(9) A, V = 2145(3) A3) shows a dinuclear structure with two mu-OH hydroxo group and two ligand molecules that are bridging two metal centers with vicinal C[double bond, length as m-dash]O groups (micro-kappa2O,O coordination mode), the octahedral geometry of the Cr atoms being completed by the coordination of the O atom of the water molecule and [Cr(micro-C5O5)(micro OH)(OH2)2]2[K5(C5O5)2.5] x 7H2O (, monoclinic, space group Cc, a = 15.216(9) A, b = 14.895(8) A, c = 18.503(9) A, beta = 96.77(5) degrees and V = 4164(4) A3), a polymer showing various coordination geometries of the K(I) cations. Magnetic susceptibility measurements, performed on , in the 5-400 K temperature range, show antiferromagnetic interactions with J1/k = -50.2 +/- 0.1 K and J2/k approximately -3 K with J1 and J2 the intradimer and interdimer interaction constants, respectively. DFT calculations, performed to gain insight into the coupling between the two metal centers and characterize their spin and charge distribution in the hydroxo-bridged dinuclear entities, are in good agreement with experimental findings. PMID- 19122915 TI - Synthetic, structural and kinetic studies on the binding of cyclohexane-1,2-bis(4 methyl-3-thiosemicarbazone) to divalent metal ions (Co, Ni, Cu, Zn or Cd). AB - The reactions of cyclohexane-1,2-bis(4-methyl-3-thiosemicarbazone) (CHMTSC) with MCl2 (M = Co, Ni, Cu or Zn) and Cd(NO3)2 have been shown to produce complexes in which the thiosemicarbazone has been doubly deprotonated [[M(CHMTSC - 2H+)] (M = Co, Ni or Ni)], analogous to those reported earlier with other Schiff base thiosemicarbazones. However, with ZnCl2 and Cd(NO3)2, the complexes isolated are [ZnCl(CHMTSC)]Cl and [Cd(NO3)(CHMTSC)]NO3, containing the protonated forms of the ligand, which have been characterised by X-ray crystallography, as has free CHMTSC. The kinetics of the reactions between CHMTSC and all the various metal salts have been determined by stopped-flow spectrophotometry. In all cases, the reactions are complete on the seconds timescale. The reactions exhibit a first order dependence on the concentration of metal salt and a first-order dependence on the concentration of CHMTSC. The thermodynamic and kinetic factors influencing the protonation state of the coordinated thiosemicarbazone are discussed. PMID- 19122916 TI - Clinical prediction of endemic rickettsioses in northern Taiwan--relevance of peripheral blood atypical lymphocytes. AB - BACKGROUND AND PURPOSE: Several rickettsioses are endemic in Taiwan. They are under-reported not only because of ignorance but also due to difficulty in recognition caused by their nonspecific manifestations, which overlap with other acute febrile illnesses. We conducted a retrospective study to delineate distinctive clinical features of rickettsiosis, in order to develop a system for differential diagnosis of rickettsiosis. METHODS: Patients admitted to Chang Gung Memorial Hospital Linkou Medical Center, Taoyuan, Taiwan, with suspected rickettsiosis during the period from January 2004 to May 2006 were included. Clinical suspicion was based on the presence of acute fever with eschar formation, relevant contact history, poor response to broad-spectrum empiric antibacterial therapy, unexplained thrombocytopenia, leukopenia, or abnormal liver biochemistry, or unexplained major organ involvement. Serum samples were sent to the Centers for Disease Control, Taiwan, for serologic diagnosis of the 3 rickettsioses endemic to Taiwan - scrub typhus (Tsutsugamushi's disease), murine typhus (endemic typhus) and Q fever. Serologically confirmed and excluded cases were compared for signs and symptoms, risk factors, laboratory findings and response to treatment. RESULTS: Among 138 suspected cases, 88 were excluded from the study because of incomplete serological tests or insufficient information, 28 were confirmed to have one of the 3 rickettsioses and 22 were negative for all of them. Distinct features among confirmed cases, compared to controls, were eschar formation, relevant contact history, and presence of atypical lymphocytes in peripheral blood. Normal or low leukocyte count, thrombocytopenia and relative bradycardia were not significant in predicting diagnosis. We propose a predictive system for tentative diagnosis of rickettsiosis based on relevant clinical attributes. This system has a positive predictive value of 80% and a negative predictive value of 100%. CONCLUSIONS: The predictive scoring system may allow institution of appropriate treatment for rickettsiosis in a more timely manner. However, a low probability of diagnosis should prompt vigorous search for other etiologies. PMID- 19122917 TI - Development of blocking ELISA for detection of antibodies against avian influenza virus of the H7 subtype. AB - BACKGROUND AND PURPOSE: The conventional method used for subtyping of antibodies against avian influenza viruses is hemagglutination inhibition (HI) test. However, the HI test is laborious and requires preparation of antigen from viable viruses that might be hazardous. The aim of this study was to develop a blocking enzyme-linked immunosorbent assay (B-ELISA) for detection of antibody of avian influenza of the H7 subtype. The B-ELISA is fast and avoids the need to culture whole viruses. METHODS: The B-ELISA was based on the reaction between a monoclonal antibody and a recombinant hemagglutinin protein purified from Escherichia coli. The specificity of the B-ELISA was determined by testing H7 negative field sera and the sensitivity of the B-ELISA was determined by testing sera collected from experimentally immunized chickens. RESULTS: The specificity of the B-ELISA was found to be 97.7% when compared with the HI test. The sensitivity was found to vary with the HI titer of sera. A sensitivity of 100% was achieved when test sera had HI titers >or=2(7). The sensitivity dropped to 33% and 20% when test sera had HI titers of 2(6) and 2(5), respectively. Nearly all test sera with HI titers 0.05). However, the prevalence of Pfmdr1 86Y was higher in those with hemoglobin SS genotype than in hemoglobin AA and AS subjects (p<0.05). CONCLUSIONS: Uncontrolled use of chloroquine is a major cause of chloroquine resistance in Nigeria. Chloroquine prophylaxis may be the underlying cause of the high prevalence of Pfmdr1 86Y mutant gene in individuals with hemoglobin SS genotype. PMID- 19122921 TI - Risk factors for mortality in patients with Acinetobacter baumannii bloodstream infection with genotypic species identification. AB - BACKGROUND AND PURPOSE: Acinetobacter baumannii is an increasingly common nosocomial infection with a high mortality rate. Identification of predictor factors of mortality from A. baumannii infection is important for the implementation of therapeutic management for patients with higher risk. However, many studies have reported data for Acinetobacter calcoaceticus-A. baumannii complex, which might lead to an uncertainty of results. In this study, we aimed to identify the predictive factors for mortality of patients infected with true A. baumannii that had been precisely identified by genotypic methodology. METHODS: Sixty seven patients with documented A. baumannii bacteremia were identified from a medical center in northern Taiwan during the period between February 1998 and February 2001. The patients' medical records were retrospectively reviewed. RESULTS: The risk factors associated with mortality in patients with A. baumannii bacteremia were underlying disease with malignancy, end-stage renal disease, and inappropriate antibiotic therapy. Laboratory variables, such as creatinine level, were also associated with poor prognosis by multivariate analysis. CONCLUSIONS: Increased serum creatinine level, malignancy and inappropriate therapy within 3 days were related to increased mortality in patients with A. baumannii bloodstream infection. Physicians should be aware of patients with poor prognostic factors and initiate prompt strategies, including appropriate antimicrobial therapy, in order to reduce mortality. PMID- 19122922 TI - Brain abscesses in children: analysis of 20 cases presenting at a medical center. AB - BACKGROUND AND PURPOSE: Brain abscess is an uncommon intracranial suppuration, especially in children. We reviewed the records of children with brain abscess admitted to Taipei Veterans General Hospital, Taipei, Taiwan, and analyzed data on presenting symptoms, predisposing factors and outcomes. METHODS: Information was obtained retrospectively from medical records of children diagnosed with brain abscess. Demographic data, presenting symptoms, predisposing factors, sites of abscess, infectious organisms, treatment regimen, and prognosis were collected and analyzed. RESULTS: Twenty cases of brain abscess within an 11-year period were included in this study. Fever, seizure and focal neurological deficits were the most common presenting symptoms. Cyanotic congenital heart disease (CCHD), present in 35% of patients, was the most common predisposing factor. Staphylococcus aureus was the most common pathogen. Approximately two-thirds of cases followed up had some form of seizure disorder sequelae; such sequelae were more common in patients with the initial presentation of seizure. CONCLUSION: CCHD was the most common predisposing factor for brain abscess in children, and a high level of seizure disorder sequelae was noted. Children suffering from brain abscess, whose initial presentation reveals seizure, are at increased risk of further seizure disorder. PMID- 19122923 TI - Campylobacter enteritis in children in northern Taiwan--a 7-year experience. AB - BACKGROUND AND PURPOSE: Campylobacter infection is recognized as a major cause of acute enteritis in humans. The clinical characteristics may vary in different countries. This study investigated the clinical manifestations of pediatric Campylobacter enteritis in a medical center in northern Taiwan. METHODS: This was a retrospective review of Campylobacter enteritis infections at National Taiwan University Hospital, Taipei, Taiwan, from January 2000 to December 2006. All children who tested positive for Campylobacter, isolated from stool samples, were included in the study. Data collected and evaluated included the species of Campylobacter, age, gender, underlying disease, travel history, clinical manifestations and laboratory data. RESULTS: A total of 104 patients had enteric campylobacteriosis. Campylobacter coli was grown from 24 patients (23.1%), while Campylobacter jejuni was found in 80 patients (76.9%). More than half of the infections (60.6%) occurred in children less than 5 years old. The male-to-female ratio was 2.46:1. Fifteen patients had underlying diseases (14.4%), such as hematologic malignancy, solid organ transplantation and liver cirrhosis. Watery diarrhea (93.2%), abdominal pain (92.0%), fever (81.2%), and vomiting (46.1%) were the most common clinical manifestations. Three episodes of campylobacteriosis appeared to be imported from Southeast Asia and 3 were acquired nosocomially. One patient, who did not have any underlying disease, developed Campylobacter bacteremia. No Guillain-Barre syndrome was noted in our patients and none of our patients died due to campylobacteriosis. While both diseases had similar clinical manifestations, infections caused by C. coli seemed to be more severe than those caused by C. jejuni, as evidenced by a higher incidence of decreased activity and pus cells in the stool in patients infected with C. coli. CONCLUSION: Even in patients with bacteremia or underlying disease, enteric campylobacteriosis usually runs a benign course regardless of treatment with antimicrobial agents in children in northern Taiwan. PMID- 19122924 TI - Adult candidemia at a medical center in northern Taiwan: a retrospective study. AB - BACKGROUND AND PURPOSE: Candidemia has been associated with a very high mortality. This study evaluated the predictors of candidemia-related mortality at a teaching hospital in northern Taiwan. METHODS: We conducted a retrospective analysis of adult patients with candidemia between September 2003 and May 2005. A stepwise logistic regression analysis was performed to determine the predictors of candidemia-associated mortality. All Candida isolates were identified to species by use of the ATB ID 32C kit and their susceptibilities to antifungal agents were tested by ATB Fungus 2 system. RESULTS: 179 episodes in 174 adult patients with candidemia were identified retrospectively. The predictors of mortality included duration of prior antibiotics >or=28 days, Acute Physiology and Chronic Health Evaluation (APACHE) II score >or=23 and retention of central venous catheters (CVCs). There was no statistically significant association between the time to the start of antifungal therapy and mortality from nosocomial candidemia. In addition, there was no significant association between the time to CVC removal and mortality after stratification by APACHE II score. CONCLUSIONS: Despite effective antifungal therapy after the onset of candidemia in more than half of the patients studied, mortality remained very high, especially in the groups with longer duration of prior antibiotic treatment, higher APACHE II score and CVC retention. Timing of CVC removal after onset of candidemia was not correlated with mortality. PMID- 19122925 TI - Comparison of CPS ID 3 and CHROMagar Orientation chromogenic agars with standard biplate technique for culture of clinical urine samples. AB - BACKGROUND AND PURPOSE: Chromogenic agars have been developed to recognize frequently occurring microorganisms directly on primary cultures, thus reducing the daily workload in a clinical microbiology laboratory. We compare two chromogenic agars, CHROMagar Orientation (CO) and CPS ID 3 (CPS3), with routine media (biplate technique using trypticase soy blood agar and eosin methylene blue agar) for the isolation, enumeration and identification of organisms in urinary tract infection (UTI). METHODS: The clinical significance of the urine samples was categorized as probable UTI, possible UTI, no UTI (negative), or contaminated according to the culture result. Discrepancy analysis with the categories of minor error, major error and very major error was used to compare the culture media. RESULTS: Of 1386 urine specimens, the consistencies in clinical significance of CO and CPS3 to routine media were 90.7% and 89.8%, respectively. For the enumeration of microorganisms, 524, 514, and 521 clinically significant isolates were isolated on routine media, CO, and CPS3, respectively. Of the 524 significant isolates on routine media, results for 473 and 474 isolates agreed on CO and CPS3, respectively. Approximately 91.9% of Escherichia coli and 100.0% of Enterococcus spp. could be identified directly on CO media, while 97.5% of E. coli and 94.4% of Enterococcus spp. could be identified on CPS3 media. CONCLUSION: The use of CO and CPS3 as single media is promising for clinical urine culture. PMID- 19122926 TI - CTX-M type beta-lactamases among fecal Escherichia coli and Klebsiella pneumoniae isolates in non-hospitalized children and adults. AB - We investigated the occurrence and diversity of extended-spectrum beta-lactamase (ESBL) enzymes among antibiotic-resistant Escherichia coli and Klebsiella pneumoniae isolates obtained from human feces. All ESBL-positive isolates were characterized at the molecular level by polymerase chain reaction, sequencing and pulsed-field gel electrophoresis (PFGE). Eight of 46 antibiotic-resistant E. coli (6 from children and 2 from adults) and 4 of 8 K. pneumoniae (all from adults) isolates were found to be ESBL-positive by the double-disk synergy test. Seven isolates were found to have CTX-M-14, 2 each had CTX-M-24 and CTX-M-38, and 1 had CTX-M-9. In addition, 8 isolates were found to carry TEM-1b or TEM-1c. No SHV type enzyme was found among the E. coli strains. In 9 strains, the plasmidic bla(CTX-M) determinants were transferable to E. coli by conjugation. Analysis by PFGE showed evidence of clonal and non-clonal spread. The present study shows fecal carriage of organisms producing bla(CTX-M) determinants and underscores the role that commensals could play as a reservoir for their dissemination. PMID- 19122927 TI - Listeria monocytogenes bacteremia in a twin pregnancy with differential outcome: fetus papyraceus and a full-term delivery. AB - A 41-year-old Saudi woman, gravida 7, para 5, aborta 1 with sickle cell disease at week 12 of gestation of diamniotic dichorionic twin pregnancy was admitted with fever. Blood cultures grew Listeria monocytogenes. She was treated with intravenous ampicillin and had death of one of the fetuses after 10 days. An ultrasound at 19 weeks of gestation showed a normal intrauterine gestation and the presence of significantly collapsed second twin gestational sac consistent with fetal papyraceus. At 37 weeks of gestation, she had a cesarean section and a full-term baby was delivered. We describe the case and review the literature on pregnancy-associated listeriosis. PMID- 19122928 TI - Erythema nodosum and hepatitis B: a case report and literature review. AB - Erythema nodosum (EN) is the most common of the panniculitides, and is associated with many underlying etiological conditions. Herein, we report a case of EN with probable association with hepatitis B virus (HBV) and review previous reports in the English literature. Since hepatitis B is still an endemic infection in Taiwan, EN related to HBV may not be as uncommon as in developed countries. The incidence and pathogenesis of EN associated with HBV demand further investigation. PMID- 19122929 TI - An unusual complication of dengue infection. AB - We present an unusual complication of dengue infection resulting in postviral phrenic neuropathy and diaphragmatic paralysis in a 34-year-old man. There is a paucity of literature on this condition, with postviral neuropathies previously reported to be associated commonly with herpes zoster, poliovirus, and rarely, West Nile virus and human immunodeficiency virus infections. To our knowledge, this is the first reported case of flavivirus causing isolated postviral phrenic neuropathy and diaphragmatic paralysis. PMID- 19122930 TI - Unusual complication of intravenous Subutex abuse: two cases of septic sacroiliitis. AB - We report two unusual cases of septic sacroiliitis resulting from intravenous Subutex abuse that initially masqueraded as low back pain. Both patients, a 48 year-old Malay man and a 30-year-old Malay woman, presented with chills, rigor and progressive lower back pain, and eventually experienced difficulty in ambulating. The Malay woman also developed severe pain in her left elbow, with swelling and restriction of movement. Blood investigations and cultures revealed an infective process. Imaging of the pelvis and lower back confirmed the diagnosis of septic arthritis of the sacroiliac joints. The first patient underwent computed tomography-guided drainage of the abscess and was administered intravenous antibiotics via a peripherally-inserted central catheter (PICC) line. The second patient underwent an arthrotomy for her elbow and her left sacroiliac joint was managed conservatively with intravenous antibiotics, also via a PICC line. The diagnostic difficulty and the need for a high index of suspicion are discussed. PMID- 19122931 TI - Persistent knee instability following revision total knee arthroplasty. AB - Instability is an important cause of failure following total knee arthroplasty (TKA). Knee stability needs to be optimal in both the coronal and sagittal planes. We describe failed revision TKA with persistent deformity and instability secondary to a deficient medial collateral ligament in a 71-year-old Chinese woman. Revision knee arthroplasty using constrained implants was performed on her with good result. During both primary and revision TKA, assessment of knee stability is critical. Constrained prosthesis would need to be considered when there is significant ligamentous deficiency. PMID- 19122932 TI - Hyperostosis and hyperphosphataemia syndrome: a diagnostic dilemma. AB - The syndrome of hyperostosis and hyperphosphataemia (HHS) is very rare. It can mimic bone infections and tumours. A nine-year-old girl presented with pain in her left lower leg. Radiographs showed patchy sclerosis in the tibial diaphysis. Investigations were normal except for hyperphosphataemia. Open biopsy showed chronic inflammation. Bacterial cultures were negative. Four months later, she had pain in the other leg. On evaluation for hyperphosphataemia, there was increased renal reabsorption of phosphates. She responded to analgesics. In patients between six and 16 years of age, HHS must be considered when there is painful diaphyseal swelling of long bones associated with isolated hyperphosphataemia. The painful episodes can recur. Surgical decompression can be considered if conservative treatment methods are ineffective. PMID- 19122933 TI - Spontaneous spinal epidural haematoma associated with aspirin intake. AB - Spontaneous spinal epidural haematoma (SSEH) is rare. No identifiable cause is found in the majority of cases, while anticoagulation accounts for up to 17 percent of cases. Aspirin-associated SSEH, however, has rarely been described in literature. A 62-year-old man on prophylactic aspirin presented with symptoms of acute cord compression due to spinal epidural haematoma that was confirmed on magnetic resonance imaging. An emergency decompression laminectomy was performed ten hours after the onset of his symptoms. No vascular anomaly was detected. Our patient recovered well and regained full motor and sensory function. Aspirin is unlikely to be the direct cause of SSEH but may predispose to it, with the underlying cause being a locus minoris resistentiae, consisting of a network of weakened epidural veins. Early diagnosis and treatment are essential for a successful outcome. PMID- 19122934 TI - Giant cavernous haemangioma of the spleen presenting as massive splenomegaly and treated by partial splenectomy. AB - Cavernous haemangioma is a rare disorder of the spleen with fewer than 100 cases reported. Only rarely do they attain large sizes. A 36-year-old woman presented with a six-month history of pain in the left hypochondrium and a massive splenomegaly. Ultrasonography, Doppler studies, and computed tomography could not distinguish between a haemangioma and a secondary deposit. Magnetic resonance imaging showed characteristic features of splenic haemangioma with central fibrosis, thrombosis and haemorrhage. Partial splenectomy was done. Intraoperative imprint cytology was negative for malignant cells. Histopathology showed cavernous haemangioma with areas of infarction necrosis. It is extremely rare to have such a massive solitary splenic haemangioma presenting as a giant splenomegaly. Preoperative investigations are often inconclusive and may not distinguish between haemangioma and metastases. Magnetic resonance imaging is the most reliable imaging method. Haemangiomas are treated only when they are symptomatic or very large with an increased risk of haemorrhage. Partial splenectomy is the treatment of choice. PMID- 19122935 TI - Adenocarcinoma of the abdominal wall. AB - We report a rare case of adenocarcinoma confined to the umbilicus. A 60-year-old menopausal woman presented with an umbilical lump of four years. Excision biopsy showed adenocarcinoma with an appearance suggestive of metastasis from an ovarian cystadenocarcinoma. Tumour markers for ovarian malignancy were normal. Computer tomography did not reveal any evidence of ovarian malignancy. She defaulted on follow-up. After five years, she presented with a 6-cm irregular periumbilical mass. She agreed to undergo an excision biopsy of the mass with total abdominal hysterectomy and bilateral salpingo-oopherectomy with omentectomy. The frozen section of the tumour showed papillary adenocarcinoma. Histology showed adenocarcinoma favouring serous cancer. There was no tumour seen in the ovaries, uterus and omentum. On follow-up, there was no disease recurrence. The patient has been disease-free for two years post surgery. PMID- 19122936 TI - Primary plasmacytoma of the uterine cervix treated with three-dimensional conformal radiotherapy. AB - Primary plasmacytoma of the uterine cervix is a rare neoplasm with limited known data, and only several cases sporadically reported in the published literature. Radiotherapy might have a role in the treatment of plasmacytoma of the uterine cervix. We describe primary plasmacytoma of the uterine cervix in a 45-year-old woman treated with three-dimensional conformal radiotherapy, and also reviewed the literature to evaluate the treatment modality and therapeutic outcome of this rare disease. PMID- 19122937 TI - A child with atypical teratoid/rhabdoid tumour of the posterior cranial fossa. AB - Primary central nervous system (CNS) atypical teratoid/rhabdoid tumours (ATRT) are highly malignant neoplasms which usually present in infancy or early childhood. Although ATRT may arise anywhere within the CNS, the majority (approximately two-thirds) arise in the cerebellum or posterior fossa, and the remainder in the cerebrum. We described the imaging characteristics of CNS ATRT in the posterior cranial fossa of a 14-month-old boy. PMID- 19122938 TI - Discordance for Ellis-van Creveld syndrome in twins. AB - Ellis-van Creveld (EVC) syndrome or chondroectodermal dysplasia is a rare autosomal recessive disorder characterised by a variable spectrum of clinical findings. Classical EVC syndrome comprises a tetrad of clinical manifestations of chondrodystrophy, polydactyly, ectodermal dysplasia, and cardiac defects. It is extremely rare for EVC syndrome to occur in one of a pair of newborn twins. Review of the existing literature revealed that only one such case has been reported so far. PMID- 19122939 TI - Supernumerary derivative (22) syndrome resulting from a maternal balanced translocation. AB - Supernumerary derivative (22) syndrome is one of the rare genomic syndromes. It is characterised by severe mental retardation, microcephaly, failure to thrive, ear anomalies, preauricular tags or sinus, cleft palate or high arch palate, microganathia, renal anomalies, congenital cardiac defects and genital abnormalities in males. In 99 percent of the cases, one of the parents is a balanced carrier of a translocation between chromosome 11 and chromosome 22. We report the first known case, a female neonate, of supernumerary derivative (22) syndrome from Malaysia. PMID- 19122940 TI - Diagnostic transgastric flexible peritoneoscopy: is pure natural orifice transluminal endoscopic surgery a fantasy? AB - We present the first transgastric peritoneoscopy in a 20-year-old man. The objectives were to evaluate the impact of the site of viscerotomy on the technical feasibility of natural orifice transluminal endoscopic surgery (NOTES), assess transgastric peritoneoscopy as a complementary procedure, determine the safety and efficacy of NOTES, and attempt inspection/biopsy of the gallbladder. The patient was admitted with a benign gastric outlet obstruction, chronic cholecystitis and radiological suspicion of a mass in the gallbladder which was not visualised on diagnostic laparoscopy. Complementary transgastric peritoneoscopy was performed to gain deeper penetration of the tumour with the flexible tip of the gastroscope. The visceral "aperture" was created in the antrum where gastrojejunal anastomosis would be fashioned. Laparoscopic transillumination of the anterior gastric wall facilitated this part of the procedure. During transgastric peritoneoscopy, the gallbladder and structures in the upper and left hemi-abdomen appeared retrograde due to the unusual location of the gastrotomy. The right hemi-abdomen and pelvis were easily examined with a "straight shaft" approach. The gallbladder could not be identified with exploratory laparoscopy and transgastric peritoneoscopy. Due to risk of visceral injury, open gastrojejunal anastomosis and cholecystectomy were performed. Intraoperatively, an inflamed, thick-walled gallbladder was found adherent to the proximal duodenum. Transgastric peritoneoscopy was safely performed in our patient. The postoperative course was uneventful. Our patient showed significant improvement at 13 weeks after surgery without any procedure-related complication. In conclusion, transgastric peritoneoscopy may be used to complement diagnostic laparoscopy. Laparoscopic assistance during transluminal access facilitates simple tasks inside the peritoneal cavity to be performed safely. PMID- 19122941 TI - Emergency department thoracotomy for pericardiac tamponade. AB - A 31-year-old man with a penetrating chest injury presented acutely with pulseless electrical activity, as a result of a ventricular laceration causing pericardial tamponade. Emergency department thoracotomy was performed to release the tamponade and he was operated on immediately to repair the laceration. He subsequently survived and was discharged to a community hospital for rehabilitation. We present this case of penetrating cardiac injury in which the patient arrived in extremis and for which emergency department thoracotomy was performed, and we discuss the role of emergency thoracotomy, its indications and outcome indicators. PMID- 19122942 TI - Aortic and mitral valve endocarditis caused by Gemella morbillorum in a haemodialysis patient. AB - Gemella morbillorum is part of the commensal flora of the upper respiratory tract, intestinal tract and genitourinary tract. On rare occasions, it causes endocarditis. We report a 67-year-old Chinese man with end-stage renal disease on maintenance intermittent haemodialysis, who developed Gemella morbillorum endocarditis complicated by severe aortic and mitral regurgitation. Most cases of Gemella morbillorum endocarditis have satisfactory outcomes with antibiotics or surgical treatment. PMID- 19122943 TI - Plagiarism: a joint statement from the Singapore Medical Journal and the Medical Journal of Malaysia. PMID- 19122944 TI - Writing the results. AB - The purpose of the results section is to present the main data collected and the observations made during the research. It provides interpretation of the analysed data and does not contain details on the methods, materials or discussion. The first step in writing the results section is to review the analysed data and determine which results to present. This can be done by deciding which results are relevant to the question(s) presented in the introduction, and may be supplemented by illustrative tables and figures. The results section guides the reader through the questions investigated in the study and sets the stage for the discussion in the next section. PMID- 19122945 TI - Appropriateness of indications for diagnostic upper gastrointestinal endoscopy in India. AB - INTRODUCTION: Guidelines for an upper gastrointestinal endoscopy have been outlined for the Western population, but not yet for India. The study aimed to assess the appropriateness of upper gastrointestinal endoscopy for patients with dyspepsia and to identify the cut-off age for endoscopy from an Indian perspective. METHODS: Patients referred for upper digestive endoscopy to a university clinic in India were prospectively studied between January 2004 and June 2005. Patients who presented with dyspepsia and those with isolated alarm symptoms without dyspepsia who underwent endoscopy were included. The cut-off age for the detection of upper gastrointestinal tract carcinoma in dyspepsia was derived. RESULTS: A total of 3,432 endoscopies were performed during the study period. There were 2,068 men and 1,364 women. The overall mean age was 41.6+/-15 (range 7-85) years. 18.3 percent of 284 patients with malignancy were between 25 and 45 years of age. Using the receiver operator characteristic curve, the cut off age for malignancy was between 35 and 44 years; specifically, the optimal cut off age was 38 years for females and 43.5 years for males. CONCLUSION: In the south Indian population with dyspepsia, there were more normal and benign lesions at endoscopy. The optimal cut-off ages for detecting malignancy for both genders were also determined. PMID- 19122947 TI - Economic cost of osteoporotic hip fractures in Singapore. AB - INTRODUCTION: The incidence of osteoporotic hip fractures in Singapore as well as in Asia will gradually increase with an ever ageing population. The objectives of this study were to understand the costs of hip fractures in our community and to analyse the various factors that lead to higher inpatient costs for hip fractures. METHODS: We prospectively reviewed 80 elderly patients with osteoporotic hip fractures. The relevant patients' clinical data was correlated with the inpatient hospitalisation costs. We reviewed the cost and management practices of hip fractures published in the literature and compared them with our findings. RESULTS: We found that our patients treated surgically incurred lower costs and had a shorter inpatient stay compared to those treated conservatively. The mean hospitalisation cost for patients treated surgically was S$10,515 and the mean length of stay was 16 days. We found that the length of stay, a longer delay to surgery, male gender, having ASA 3 score, and development of postoperative complications led to higher inpatient hospital costs. CONCLUSION: With the identification of various clinical factors that are associated with high inpatient costs, we can further shorten our hip fracture patients' stay as well as reduce the hospitalisation costs. PMID- 19122946 TI - Increased mast cell density in haemorrhoid venous blood vessels suggests a role in pathogenesis. AB - INTRODUCTION: Haemorrhoids are an abnormal, tortuous dilatation of the arteriovenous plexus of the anus. Although increased resting anorectal pressure is deemed to be a major initiating factor, a thorough understanding of the pathogenesis is still lacking. Mast cells, through release of granules, can affect local vessels with respect to changes in calibre, changes in permeability and thrombosis. Thus, mast cells could play a role in haemorrhoid pathophysiology, although this has not been previously investigated. METHODS: 48 cases of haemorrhoids were retrospectively collected at King Chulalongkorn Memorial Hospital, with normal anorectal tissue from surgically-removed colorectal cancer serving as controls. Mast cells were identified by toluidine blue staining and quantitated around venous vessels. RESULTS: Mast cells around haemorrhoidal vessels were significantly more numerous than in normal specimens (p-value is less than 0.001). Similar values were found for haemorrhoids showing chronic changes and those in a more acute stage. CONCLUSION: These findings support the hypothesis that mast cells may play a role in the pathophysiology of haemorrhoids. Mast cells appear to participate equally in the early and later stages of these lesions. Mast cells are known to affect local vascular conditions through release of their chemical mediators and cytokines, and may influence haemorrhoid symptomatology and progression at this level. PMID- 19122948 TI - Knowledge of primary paediatric care providers regarding attention deficit hyperactivity disorder and learning disorder: a study from Pakistan. AB - INTRODUCTION: Attention deficit hyperactivity disorder (ADHD) and learning disorder (LD) remain prevalent globally and are also speculated to have a high occurrence in Pakistan. An early diagnosis and intervention in these disabilities is imperative for achieving good clinical and functional outcomes. This can be ensured by an effective screening at the level of primary paediatric care in the developing countries. We aimed to explore the ability of general practitioners (GPs) and paediatricians in Pakistan to screen for ADHD and LD based on their awareness regarding the risk factors and symptomatology of ADHD and LD. METHODS: A total of 96 paediatricians and 98 GPs practising in Karachi, Pakistan were included in the study. Data was collected employing a self-administered questionnaire. RESULTS: Only 13.7 percent of the GPs and 21.6 percent of the paediatricians were shown to have knowledge sufficient to effectively screen for/diagnose ADHD. Alarmingly, not a single GP was adequately familiar with the established risk factors and clinical symptoms of LD. The level of knowledge was not influenced by age, gender, and clinical practice attributes of the physicians. Doctors who regularly read medical journals and attend medical education seminars showed slightly better trends. CONCLUSION: We hypothesise that this demonstrated lack of knowledge at the level of primary care in Pakistan prevents an early screening of ADHD and LD. A multipronged strategy targeted at the provision of objective screening tools for primary paediatric care providers, regular continuing medical education seminars and an emphasis on paediatric mental health in undergraduate and postgraduate curricula may ensure an early detection of ADHD and LD in Pakistan. PMID- 19122949 TI - Laparoscopic appendicectomy for complicated appendicitis in children. AB - INTRODUCTION: The place of laparoscopic appendicectomy in the management of complicated appendicitis remains unsettled with reports of a higher incidence of postoperative intraperitoneal abscess. Most studies on laparoscopic appendicectomy in children have been done in the Western population. This retrospective review was done to compare laparoscopic appendicectomy with open appendicectomy in children with complicated appendicitis in a hospital in Malaysia. METHODS: The laparoscopic appendicectomies were done by a single surgeon, while the open appendicectomies were performed by surgical trainees with at least three years of surgical experience. There was no selection of cases for laparoscopic appendicectomy. Both procedures were done by standard techniques. The operative time, duration of postoperative stay, wound infection rate, incidence of intraperitoneal abscess and postoperative fever were compared. RESULTS: Based on the intention to treat analysis, there were 51 and 61 children in the laparoscopic and open appendicectomy groups, respectively. Conversion from laparoscopy to open technique was done in six children. Distribution of age, gender and pathology of appendicitis was similar for the two groups. With laparoscopic appendicectomy, the mean operative time was longer (112 vs. 72 minutes, p-value is less than 0.005), while severe wound infection (2.1 vs. 21 percent, p-value is less than 0.05) and mean postoperative hospital stay (5.7 vs. 7.4 days, p-value is less than 0.005) were significantly lower. The incidence of postoperative intraperitoneal abscess (12 vs. 11.5 percent) and postoperative fever (20 vs. 11.5 percent) were not significantly different (p-value is greater than 0.05). CONCLUSION: This review has confirmed that the longer the operating time, the lower the incidence of wound infection and the shorter the postoperative stay for laparascopic appendicectomy. There was no increased risk of postoperative intraperitoneal abscess. PMID- 19122950 TI - Palliative radiotherapy in paediatric malignancies. AB - INTRODUCTION: Providing effective palliative treatment in childhood malignancies is a challenging task. This study evaluated the role of palliative radiotherapy in the management of incurable paediatric malignancies. METHODS: Records of 40 paediatric patients treated between January 2003 and November 2005 were reviewed and analysed retrospectively. All had received palliative external beam radiotherapy for symptom control either as a single modality or in addition to surgery, chemotherapy and drugs for symptomatic relief. RESULTS: Predominant symptoms noticed were swelling with or without pain, bleeding, and weakness of limbs. Median duration of symptoms was 90 days. Malignant round cell tumours were most common followed by retinoblastoma, neuroblastoma, Ewing's sarcoma and acute myeloid leukaemia with chloromas. 45 percent of children had disseminated disease at presentation. Nine underwent surgery, while 32 patients received chemotherapy, and all but two received drugs for symptomatic relief in addition to palliative radiotherapy. Dose schedules were either 5 Gy or 8 Gy in single fraction, while for fractionated radiotherapy, the range was 20 Gy in five fractions to 30 Gy in ten fractions. With regard to symptomatic relief, four patients had complete relief, 20 showed good relief, 15 had little and one did not have any relief. On completion of multimodality treatment, tumour response was complete in two patients, 18 had partial response, eight had stable disease, eight had progressive disease, and the disease status of four was unknown. CONCLUSION: The role of radiotherapy as a palliative modality in children with locally-advanced lesions provides better symptomatic relief in combination with other treatment modalities. PMID- 19122951 TI - Breast-feeding patterns and factors determining exclusive breast-feeding. AB - INTRODUCTION: Breast-feeding is a major determinant of infants' health and survival. The aim of this study was to examine breast-feeding patterns and the role of some factors on exclusive breast-feeding. METHODS: The study was done at the Health Centre of Community Oriented Medical Education of Zanjan City, Iran. Mothers of 650 infants younger than one year of age were interviewed directly by healthcare providers. The necessary information was recorded on pre-structured questionnaires, and the feeding patterns of all infants were determined. RESULTS: The rate of breast-feeding was greater than 92 percent during the first year of life. Exclusive breast-feeding rate in the first five days of life was 82 percent, but it declined to 44 percent at the first month. The chance of exclusive breast-feeding was higher in infants who were on demand feeding (odds ratio [OR] 2.6), breastfed more than eight times per day (OR 2.23), and their birth weight ranged from 2.5 to 4 kg (OR 2.16). This chance was less among infants exposed to pacifiers (OR 2.62) and with the numbers of deliveries equal to or more than three (OR 2.25). CONCLUSION: The rates of breast-feeding initiation and its duration at six and 12 months were higher than the national goal. The rate of exclusive breast-feeding was nearly 44 percent. It seems necessary to elucidate the role of other factors, such as socioeconomic status, on exclusive breast-feeding. PMID- 19122952 TI - The acute pain service after ten years: experiences of a Singapore public hospital. AB - INTRODUCTION: The management of postoperative pain is an increasingly important aspect of healthcare, leading to the establishment of acute pain services in major surgical centres worldwide. The acute pain service (APS) was established in most Singapore hospitals in the 1990s. We analysed data collected in our institution over a ten-year period (1998-2007), documenting our experiences, outcomes and complications encountered by our APS. METHODS: Data was chronologically divided into two groups for analysis: years 1998-2003 (3,248 cases) and 2004-2007 (2,466 cases). Analysis included a comparison of patient profiles, modalities of analgesia used, effectiveness of pain relief, adverse effects, complications and patient satisfaction. Results were also compared to published audits and proposed standards in medical literature. RESULTS: The patient profile served by the APS remained unchanged over the years, but a move away from central neuraxial blocks was noted with an increased utilisation of patient-controlled analgesia. There was no clinically significant change in pain scores over the two periods of analysis (0.9 vs. 1.0 at rest, 3.0 vs. 3.0 on movement). There were also no statistically significant changes in the prevalence of patients reporting severe pain while on the APS (1.5 percent vs. 1.6 percent at rest, p-value is 0.66; 8.5 percent vs. 9.4 percent on movement, p-value is 0.25). Complication rates remained well within international standards and no major complications were reported. Patient satisfaction remained high (94.3 percent vs. 94.6 percent, p-value is 0.6). CONCLUSION: The move away from invasive and less targeted analgesic modalities has not compromised the quality of analgesia provided. Major morbidity remains extremely rare and incidence of complications has been reduced over the years. Patient satisfaction remains well in excess of 90 percent, and the side effects are largely well controlled. With further advancements in the provision of acute postoperative analgesia, the APS will continue to play an important role in the holistic convalescence of the surgical patient. PMID- 19122953 TI - Prevalence and predictive factors for complementary and alternative medicine use in Brunei Darussalam. AB - INTRODUCTION: Complementary and alternative medicine (CAM) use is common among patients with chronic disorders. CAM use is also reported to be common among the general population. This cross-sectional study assessed the prevalence, types of ingested CAM use and the factors predictive of their use in Brunei Darussalam. METHODS: 568 visitors (male 41.4 percent, mean age 40.2+/-15.1 years) to the medical wards were randomly approached and invited to participate in this study. Demographical data (gender, age and racial groups), comorbidities, smoking history, use of prescribed medications and psychosomatic symptoms of depression (anxiety, backache, depression, headache and insomnia) and CAM use were collected. RESULTS: Overall, 21.1 percent had used CAM in the last 12 months, comprising traditional remedies (35.8 percent), vitamins/minerals supplements (19.2 percent), non-vitamins/non-minerals supplements (30 percent) and herbal remedies (3.3 percent). 11.7 percent were unsure of the CAM they had taken. On univariate analysis, older age (40 years and above) and presence of psychosomatic symptoms of depression, particularly backaches, depression, headache and insomnia, were predictive for CAM use. On multivariate analysis, only the presence of any psychosomatic symptoms of depression remained significant for CAM use (odds ratio 1.7, 95 percent confidence interval 1.1-2.6). Among those using prescribed medications, 21.9 percent were also using CAM. 21 percent of subjects without any comorbidities reported using CAM. There was no report of any adverse events. CONCLUSION: CAM use is not uncommon. One-fifth of our subjects have used CAM within the past 12 months, even in those who do not have any comorbidity. Presence of any psychosomatic symptom of depression was predictive for CAM use. Most importantly, a proportion of the users were not even sure of what they had taken. PMID- 19122954 TI - Significance of the 'ovarian crescent sign' in the evaluation of adnexal masses. AB - INTRODUCTION: This paper aimed to study the ability of the "ovarian crescent sign" to predict the nature of adnexal masses and to validate the "ovarian crescent" as an ultrasonographical marker for malignancy. METHODS: A prospective study was carried out in 60 consenting women with an undiagnosed adnexal mass, attending the gynaecology service and requiring operative intervention. An ovarian crescent sign at pelvic ultrasonography was considered to be present if normal ovarian tissue was seen adjacent to the tumour area. The ultrasonographer was blinded to the reports of CA 125, and if applicable, the ascitic fluid cytology and needle aspiration biopsy. Histopathological examination report of the tumour obtained at surgery (laparotomy/laparoscopy) was considered as the gold standard. RESULTS: 11 of 60 biopsy specimens were positive for malignancy. Normal ovarian tissue could be identified (positive crescent sign) in nearly two thirds of cases (65 percent) scanned. Presence of normal ovarian tissue was identified in 97 percent of the benign masses. The sign was not seen in ten of the 11 cases with malignancy. CONCLUSION: The ovarian crescent sign as a method in prejudging the adnexal masses was found to have high sensitivity (90.9 percent) and high negative predictive value (97.4 percent). PMID- 19122955 TI - Comparative study of the impact of past pregnancy outcome on future fertility. AB - INTRODUCTION: The impact of previous pregnancy outcome on future fertility in Nigeria has not been appreciated, hence the continued neglect of the adoption of safe motherhood measures in this sub-region. The objective of this study was to compare the past pregnancy outcomes among fertile and infertile patients in a Nigerian population. METHODS: An institutional-based comparative case-controlled study of past pregnancy outcomes among infertile and fertile women was conducted. The data was managed using Epi-Info and the Statistical Package for Social Sciences. RESULTS: 708 patients consisting of 472 pregnant women (fertile) and 236 infertile women were investigated. Infertile women were at a significant risk of having an adverse pregnancy outcome, such as induced abortion (p-value is 0.0001), postabortal sepsis (p-value is 0.0001), postpartum infection (p-value is 0.001), manual removal of the placenta (p-value is 0.0005) and prolonged unsupervised labour (p-value 0.0001), compared to pregnant fertile women. Logistic regression analysis of variables at 95 percent confidence intervals showed that the adjusted odds ratio for prolonged labour, prolonged rupture of membranes, postabortal sepsis and postpartum infection still remained significant. CONCLUSION: Previous mismanaged pregnancies impacted negatively on future fertility. Efforts should be directed towards the prevention of unplanned pregnancies. Motherhood must be made safer in planned pregnancies by prevention of complications, and aggressive and prompt treatment of any complication if and when it occurs. PMID- 19122956 TI - Refining quantitative fluorescent polymerase chain reaction for prenatal detection of X chromosomal anomalies in the major Southeast Asian populations. AB - INTRODUCTION: This study aimed to refine the current quantitative fluorescent polymerase chain reaction (QF-PCR) screen to detect X chromosome anomalies for prenatal diagnosis in the major Southeast-Asian populations. METHODS: 100 amniotic fluid samples from Chinese, Malay and Indian origins were subjected to QF-PCR using the X chromosome markers, HPRT, X22 and AMXY, along with the autosomal marker D21S1411. RESULTS: Out of the 100 samples tested by markers X22 and HPRT, eight samples were homozygous for both markers, of which seven were resolved by comparison with the autosomal marker D21S1411. CONCLUSION: 99 percent of samples could be tested for X chromosome copy numbers, increasing the stringency for detection of X chromosome anomalies by QF-PCR. All results were confirmed by cytogenetics. PMID- 19122957 TI - Chiari 1 malformations: an Indian hospital experience. AB - INTRODUCTION: A consensus on the ideal surgical procedure for Chiari 1 malformation has not been achieved. The purposes of the study were to report on the experience of treating this condition in an Indian hospital, and to look into the management of this not too uncommon condition with a view to improve the treatment and prognosis. METHODS: A retrospective case note study of all cases of Chiari 1 malformations operated over a ten-year period from 1989 to 1999 and followed-up for another two years, were analysed for clinical and radiological features. All patients who underwent posterior fossa decompression were included. Patients who underwent other procedures like shunts were excluded. The data obtained included clinical and radiological features. RESULTS: A total of 51 cases were analysed. All patients had posterior fossa decompression with duroplasty in the majority. Many patients had a delayed improvement noticed during the follow-up period. Headache, neck pain and motor features showed an improvement in the majority of the patients but sensory deficits did not. CONCLUSION: This study showed the effectiveness of posterior fossa decompression with duroplasty as a procedure, which benefited most patients with Chiari 1 malformations. PMID- 19122958 TI - Efficacy of 80% phenol solution in comparison with cryotherapy in the treatment of common warts of hands. AB - INTRODUCTION: The common wart is a common infectious disease caused by human papilloma virus. A variety of therapeutic modalities are available. Cryotherapy (liquid nitrogen) is one of the most common treatment forms. It freezes the tissue and destroys warts. Phenol is a caustic agent. Our purpose was to evaluate and compare the efficacy of cryotherapy and 80 percent phenol solution on common warts of hands. METHODS: This single-blinded clinical trial study was performed on 60 patients with common warts referred to the dermatology clinic of Ghaem Hospital Mashhad, Iran, in 2002. Patients were randomly divided into two groups; 30 patients were treated with cryotherapy and 30 patients were treated with 80 percent phenol, on a once-weekly basis until complete clearance of the lesions or a maximum duration of six weeks. RESULTS: Complete clearance of warts after six weeks was observed in 70 percent of patients who were treated with cryotherapy, and 82.6 percent of patients in the 80 percent phenol group; there was no statistically significant difference between the two methods (p-value is 0.014). CONCLUSION: Our data indicates that 80 percent phenol and cryotherapy are effective and simple treatments for common warts of hands, and patients do not experience any pain during the treatment. PMID- 19122959 TI - Reliability of a Malay-translated questionnaire for use in a hand-arm vibration syndrome study in Malaysia. AB - INTRODUCTION: Validity and reliability of the information relating to hand transmitted vibration exposure and vibration-related health outcome are very important for case finding in hand-arm vibration syndrome (HAVS) studies. In a local HAVS study among a group of construction workers in Kuala Lumpur, Malaysia, a questionnaire translated into Malay was created based on the Hand-transmitted Vibration Health Surveillance--Initial Questionnaire and Clinical Assessment, from Vibration Injury Network. This study was conducted to determine the reliability of standardised questions in the questionnaire used in the study. METHODS: 15 subjects were selected randomly from the sampling frame of the HAVS study. Test-retest reliability was conducted on all items contained in parts 1-6 of the questionnaire and clinical assessment form, with an interval of 13-14 days between the first and second administration. Kappa coefficient and percentage agreement were calculated for all standardised questions. RESULTS: The kappa coefficient and percentage agreement for all standardised questions varied from 0.174 to 1.000 and 66.7 to 100.0 percent, respectively. The kappa coefficient for important questions related to current vibratory tool usage, tingling, numbness and hand grip weakness were 0.714, 0.432, -0.077 and -0.120, respectively, while the percentage agreement for current vibratory tool usage, finger colour change, tingling, numbness and hand grip weakness were 85.7 percent, 92.8 percent, 79.5 percent, 85.7 percent and 71.4 percent, respectively. Intra-rater reliability on the extent of vibration exposure was good, with the intra-class correlation coefficient (95 percent confidence interval) ranging from 0.786 (0.334-0.931) to 0.975 (0.923-0.992). CONCLUSION: Critical questions on vascular, neurological and musculoskeletal symptoms of HAVS were found to be reliable. The history on the extent of vibration exposure revealed good reliability when explored by the investigator alone. This questionnaire is considered reliable to be used in the study of HAVS among construction workers working in a construction site. PMID- 19122960 TI - Detection of beta-globin gene mutations among Kelantan Malay thalassaemia patients by polymerase chain reaction restriction fragment length polymorphism. AB - INTRODUCTION: Beta-thalassaemia major is an autosomal recessive disorder that results in severe microcytic, hypochromic, haemolytic anaemia among affected patients. Beta-thalassaemia has emerged as one of the most common public health problems in Malaysia, particularly among Malaysian Chinese and Malays. This study aimed to observe the spectrum of mutations found in Kelantan Malay beta thalassaemia major patients who attended the Paediatrics Daycare Unit, Hospital Universiti Sains Malaysia, Kelantan, Malaysia, the data of which was being used in establishing the prenatal diagnosis in this Human Genome Centre. METHODS: This was a cross-sectional study conducted with 35 Kelantan Malay beta-thalassaemia major patients. DNA was extracted from the blood collected from the patients and subjected to polymerase chain reaction (PCR) amplification. Six restriction enzymes were used to digest the PCR products for the detection of mutations. RESULTS: Five out of the six beta-globin gene defects were detected, namely, IVS 1 nt5 (G>C), IVS-1 nt1 (G>T), codon 26 (G>A), codon 41-42 (4 bp del) and codon 19 (A>G). The mutation which was not observed in this study was in codon 15 (G>A). The two most common mutations observed were codon 26 (G>A) and IVS-1 nt5 (G>C), which was detected in 26 and 17 patients, respectively. Two patients did not show any of the six mutations. CONCLUSION: Our results added to the existing data on the common beta-globin gene defects in Kelantan Malay beta-thalassaemia patients. PMID- 19122961 TI - Disease mongering: the role of medical journals. PMID- 19122963 TI - Use of psychoactive drugs among patients visiting outpatient clinics in Karachi, Pakistan. PMID- 19122965 TI - Clinics in diagnostic imaging (125). Diagnosis: Suspicious right breast carcinoma with axillary node metastases. AB - A 61-year-old woman who had a known history of ovarian carcinoma presented with a palpable painless mass in the right axilla. Mammograms showed segmental distributed pleomorphic microcalcifications in the upper outer quadrant of the right breast with marked enlargement of the right axillary nodes. The biggest node contained microcalcifications. Right axillary node dissection showed metastatic adenocarcinoma which was likely to be metastasis from the primary breast carcinoma. Unfortunately, she was then lost to follow-up and came back again with a right breast mass. Histopathology of the right breast mass revealed invasive ductal carcinoma. The causes and differential diagnosis of axillary adenopathy are discussed. In a patient with known primary extramammary malignancy and axillary adenopathy, it is important to differentiate if it is metastasis from the primary breast carcinoma or extramammary malignancy to provide proper management. PMID- 19122971 TI - DNA-induced aggregation and fusion of phosphatidylcholine liposomes in the presence of multivalent cations observed by the cryo-TEM technique. AB - By means of cryoelectron transmission microscopy (cryo-TEM), we were able to demonstrate the formation of ternary complexes (TC): DNA-phosphatidylcholine liposome-divalent metal cations. Addition of Ba2+ to TC led to visualization of DNA compacting on the liposome surface. Staining the TC by Tb3+ cations revealed the changed secondary structure of DNA located between fused liposomes. Cryo-TEM and liposome turbidity data were analyzed during TC formation. Liposome aggregation and the liposome fusion induced by DNA in TC were observed. Because TC displayed the property of DNA cationic liposome complexes as well as their own unique properties, we were able to consider cationic lipoplexes as a particular case of TC. The involvement of TC and direct DNA-lipid interactions in the formation nuclear pore complexes were assumed. PMID- 19122972 TI - Effects of lipids on ENaC activity in cultured mouse cortical collecting duct cells. AB - Direct effects on epithelial Na+ channels (ENaC) activity by lipids, e.g., arachidonic acid (AA), eicosatetraynoic acid (ETYA), linoleic acid (LA), stearic acid (SA), hydroxyeicosatetraenoic acid (HETE), 11,12-epoxyeicosatrienoic acid (EET), (PGF2), and (PGE2), in cultured mouse cortical collecting duct (M1) cells were clarified by using single-channel recordings in this study. In a cell attached recording, a bath application of 10 microM AA significantly reduced the ENaC open probability (NPo), whereas 10 microM ETYA or 5 microM LA only induced a slight inhibition. The inside-out recording as a standard protocol was thereafter performed to examine effects of these lipids on ENaC activity. Within 10 min after the formation of the inside-out configuration, the NPo of ENaC in cultured mouse cortical collecting duct (M1) cells remained relatively constant. Application of ETYA or LA or SA exhibited a similar inhibition on the channel NPo when applied to the extracellular side, suggesting that fatty acids could exert a nonspecific inhibition on ENaC activity. 11,12-EET, a metabolite of AA via the cytochrome P450 epoxygenase pathway, significantly inhibited the ENaC NPo, whereas 20-HETE, a metabolite of AA via the hydroxylase pathway, only caused a small inhibition of the ENaC NPo, to a similar degree as that seen with ETYA and LA. However, both PGE2 and PGF2alpha significantly enhanced the ENaC NPo. These results suggest that fatty acids exert a nonspecific effect on ENaC activity due to the interaction between the channel proximity and the lipid. The opposite effects of 11,12-EET and prostaglandin (PG) implicate different mechanisms in regulation of ENaC activity by activation of epoxygenase and cyclooxygenase. PMID- 19122973 TI - Impedance spectroscopic investigation of the bilayer lipid membranes formed from the phosphatidylserine-ceramide mixture. AB - Electrochemical impedance spectroscopy was used for the study of two-component lipid membranes. Phosphatidylserine and ceramide were to be investigated because they play an important biochemical role in cell membranes. The research on biolipid interaction was focused on a quantitative description of processes that take part in a bilayer. Assumed models of interaction between amphiphilic molecules and the equilibria that take place there were described by mathematical equations for the studied system. The possibility of complex formation for a two component system forming bilayers was assumed, which could explain the deviation from the additivity rule. The molecular area and the equilibrium constant of the complex were determined. PMID- 19122975 TI - Determining important regulatory relations of amino acids from dynamic network analysis of plasma amino acids. AB - The changes in the concentrations of plasma amino acids do not always follow the flow-based metabolic pathway network. We have previously shown that there is a control-based network structure among plasma amino acids besides the metabolic pathway map. Based on this network structure, in this study, we performed dynamic analysis using time-course data of the plasma samples of rats fed single essential amino acid deficient diet. Using S-system model (conceptual mathematical model represented by power-law formalism), we inferred the dynamic network structure which reproduces the actual time-courses within the error allowance of 13.17%. By performing sensitivity analysis, three of the most dominant relations in this network were selected; the control paths from leucine to valine, from methionine to threonine, and from leucine to isoleucine. This result is in good agreement with the biological knowledge regarding branched chain amino acids, and suggests the biological importance of the effect from methionine to threonine. PMID- 19122974 TI - Involvement of amino acid 36 in TM1 in voltage sensitivity in mouse Na+/glucose cotransporter SGLT1. AB - Human SGLT1 protein is an established sodium-glucose cotransporter. Despite widespread use of the mouse as a model organism, the mouse SGLT1 homologue has yet to be functionally characterized. Additionally, the crystal structure of a sugar transporter homologue, Vibrio SGLT, has recently been described, however, it offers limited information about the role of transmembrane segments outside of the core ligand binding domains. In particular, the amino acids in TM1 were not assigned in the structure. To examine the contribution of TM1 to the function of SGLT1, we have cloned and characterized the biophysical properties of SGLT1 from mouse, mSGLT1, and compared it to a clone containing an amino acid substitution in TM1, F36S. As predicted, both proteins formed functional Na+/sugar cotransporters, but F36S-mSGLT1 showed decreased rates of sugar uptake and decreased apparent affinities for both Na+ and sugar compared to mSGLT1. Analysis of pre-steady-state currents and comparison with the crystal structure of Vibrio SGLT provide plausible mechanisms to explain the differences in function of these two proteins. Our data suggest that amino acids in TM1, which are not involved in ligand binding and translocation pathways, significantly influence the functional properties of sodium-glucose carrier proteins. PMID- 19122976 TI - Comparing the cost of adjuvant anastrozole with the benefits of managing less patients with relapsed breast cancer. AB - Prescribing anastrozole instead of tamoxifen increases initial adjuvant drug costs but there is an eventual saving as fewer patients will relapse. The effect of this saving depends on an accurate understanding of the cost of breast cancer relapse. We identified 232 patients relapsing between March 2000 and 2005. Seventy-seven were randomly selected for analysis of their entire hospital and community management costs from the date of relapse until death, or the end of the evaluation period (01/01/07). The mean cost per patient was 25,186 pounds (95% CI 13,705 pounds- 33,821 pounds). The median survival from time of relapse was 40.07 months (range 0.5-73 months) and median total cost per patient was 31,402.62 pounds. Equating this figure with the difference in relapse rate (4.1%), initial drug cost (4,773 pounds) gives an extra cost of 17,244 pounds/life year saved. This was the first adjuvant cost effectiveness analysis which included the community management activity of a subsequent relapse. PMID- 19122977 TI - Aspirin and other non-steroidal anti-inflammatory drugs and risk of colorectal cancer: a Danish cohort study. AB - OBJECTIVE: The optimal duration and dose of aspirin and non-aspirin non-steroidal anti-inflammatory drugs (NSAIDs) in the potential prevention of colorectal cancer (CRC) have not been established. We examined this issue in the Danish Diet, Cancer, and Health Study. METHODS: Self-reported NSAID use at entry (January 1995 May 1997) was updated through June 2006, using a nationwide prescription database. CRC incidence was ascertained from nationwide registers. Cox proportional hazards regression was used to compute confounder-adjusted incidence rate ratios (RRs) and 95% confidence intervals (CIs). RESULTS: From 51,053 cohort subjects, we identified 615 CRC cases during 1995-2006. Daily aspirin use at entry was associated with a decreased risk of CRC (RR, 0.73; 95% CI, 0.49-1.10). A similar risk reduction was seen among subjects with 10 or more prescriptions for aspirin or non-aspirin NSAIDs and five or more years of follow-up. Most aspirin prescriptions were for 75-150 mg aspirin tablets. Among non-aspirin NSAID users, only those with body mass index (BMI) above 25 showed risk reductions [RR, 0.69 (0.47-1.03) for 10 or more prescriptions]. CONCLUSIONS: Long-term consistent use of aspirin or non-aspirin NSAIDs appears necessary to achieve a protective effect against CRC. Further studies of the effective dose of aspirin and the potential interaction between NSAID use and BMI are warranted. PMID- 19122980 TI - Polypharmacy, inappropriate prescribing and adverse drug reactions in Austria. PMID- 19122981 TI - [Medicine between science and art]. PMID- 19122983 TI - Soft tissue sarcoma. PMID- 19122982 TI - [Radiotherapy of soft tissue sarcoma--part of a multidisciplinary strategy]. AB - The management of soft tissue sarcoma has evolved from a solitary surgical treatment to an interdisciplinary multimodal approach including radiotherapy. These fundamental changes are the result of increased knowledge in tumor biology, radiation sensitivity and the improvement in modern radiation therapy techniques. A successful effective therapy regimen strongly depends on distinct preoperative diagnostics, preoperative conception of the surgical intervention and an experienced oncological team. Of significant importance for the prognosis is early diagnosis as well as tumor excision with a wide negative margin. However, even after complete wide resection in sano, the use of postoperative radiotherapy can further improve local control and should therefore be applied to the majority of patients. Consequently, radiotherapy should only be omitted in cases in which the tumor has been excised with a very wide negative margin; this implies, however, high quality of surgery and distinct histopathological analysis. Patients with non- or questionable resectable tumors, should be referred for pre operative radiotherapy in order to improve the surgical results. Recent studies have underlined the efficiency of modern radiotherapy regimens. The different radiotherapy regimens will be highlighted against the background of tumor stage and tumor resectibility. PMID- 19122984 TI - Polypharmacy and inappropriate prescribing in elderly internal-medicine patients in Austria. AB - OBJECTIVE: The aim of the study was to assess the prevalence of polypharmacy and inappropriate drug use in elderly internal-medicine patients in one Austrian center and to define the impact of these and other identified predictors on the occurrence of adverse drug events. METHODS: All patients>or=75 years admitted to selected internal wards of a university hospital were included in a monocentric prospective cohort study over a period of three months. The pre-admission medication of the patients was analyzed with respect to appropriateness by a multidisciplinary team consisting of pharmacists and physicians trained in internal medicine. The medication was evaluated for the occurrence of adverse drug events. RESULTS: A total of 543 patients were analyzed (median age 82 years; 60.2% female). The mean number of drugs taken was 7.5+/-3.8, with women taking significantly more drugs than men (7.8 vs. 6.8, P=0.013). Overall, 58.4% of the patients fulfilled the given criteria for polypharmacy (>6 drugs). The following factors were associated with polypharmacy: female sex, need for nursing care, high number of discharge diagnoses and high Charlson comorbidity score. Unnecessary drugs were found prescribed in 36.3% of all patients, drugs to avoid (Beers criteria) in 30.1%, duplication in 7.6%, wrong dosage in 23.4% and possible drug-drug interactions in 65.8%. Adverse drug events were identified in 17.8% of the patients (97/543), among whom the adverse drug event was the reason for hospital admission in 56.7% of the cases and a drug-drug interaction was involved in 18.7%. Risk factors for adverse drug events were female sex, polymorbidity, renal dysfunction and inappropriate prescribing. CONCLUSION: Polypharmacy, inappropriate prescribing and adverse drug events were highly prevalent in a cohort of elderly internal-medicine patients in Austria. To improve drug safety in this high-risk population, appropriate prescribing might be more important than simply reducing the number of prescribed drugs. PMID- 19122985 TI - Prevention of contrast media-induced nephropathy by isotonic sodium bicarbonate: a meta-analysis. AB - BACKGROUND: The clinical value of hydration therapy using sodium bicarbonate in the prevention of contrast media-induced nephropathy is subject of controversy. OBJECTIVE: This meta-analysis of randomized clinical trials was performed to determine whether hydration therapy using sodium bicarbonate consistently differs from control hydration regimens with normal saline in its preventive effects upon radiocontrast media-induced renal dysfunction. METHODS: Randomized clinical trials comparing sodium bicarbonate with a control regimen for prevention of contrast media-induced nephropathy were selected by multiple methods, including computer searches of bibliographic databases, perusal of reference lists, and manual searching. Major findings were qualitatively summarized. In addition, a quantitative meta-analysis was performed on renal dysfunction data. RESULTS: Altogether, 9 randomized clinical trials with a total of 2,043 patients were included. The pooled odds ratio was 0.45 (confidence interval, 0.26-0.79), indicating a significant treatment benefit of sodium bicarbonate compared to normal saline. Heterogeneity was detectable (p=0.016) and evidence of publication bias was present (p=0.012). After adjustment for presumed unpublished trials after trim-and-fill analysis, the estimated pooled odds ratio was 0.65 (confidence interval, 0.36-1.20). CONCLUSION: This meta-analysis confirms that, based on currently available randomized trials, bicarbonate therapy is effective in preventing contrast media-induced nephropathy. However, study heterogeneity and publication bias are substantial, and therefore further controlled clinical trials are needed. PMID- 19122987 TI - Is day-case cataract surgery an attractive alternative from the patients' point of view? A questionnaire survey. AB - Cataract surgery is the most common elective surgical procedure undertaken in elderly people. In many European countries and in the USA cataract surgery is normally a day-case procedure without an overnight stay in hospital, unlike the situation in Austria where fewer than 2% of patients are day cases. However, there is a lack of prospective studies on patients' need for and acceptance of day-case surgery. The aim of our study was therefore to evaluate patients' demand and suitability for outpatient surgery, based on analysis of preoperative questionnaires on availability of family and/or social support and on preoperative ophthalmologic examination. Among 500 consecutive patients with cataract, 154 (41.8%) chose a day-case procedure and 256 (58.2%) preferred inpatient admission. Patients preferring full admission were older (mean age 76.4+/-8.1 vs. 72.73+/-9.5 years, P<0.001), had worse visual acuity (0.55+/-0.20 vs. 0.66+/-0.23, P<0.001), were more likely to be female (63.9% vs. 47.0%, P=0.001), to live more than one hour away from the hospital (26.5% vs. 12.5%, P<0.001) and have no carer at home (43.0% vs. 6.5%, P<0.001). Patients favoring day-case surgery were more likely to be able to attend an ophthalmologic check-up 24 h after surgery (96.7% vs. 59.6%, p<0.001). The majority of patients indicated a preference for inpatient care, but this may be a reflection of their previous hospital experience and a matter of custom in the elderly population. Information, education and better organization of after-surgery services could help increase the attractiveness of cataract surgery as an outpatient procedure in Austria. Our findings could also be relevant to day-case services in medical care in general. PMID- 19122986 TI - Abnormal pulmonary arterial pressure limits exercise capacity in patients with COPD. AB - OBJECTIVE: Pulmonary hypertension (PH) is common in patients with chronic obstructive pulmonary disease (COPD). Mean pulmonary artery pressure (mPAP) is often only slightly elevated at rest but is increased by exercise. The purpose of this study was to determine whether abnormal pulmonary artery pressure impairs exercise capacity in patients with COPD. PATIENTS AND METHODS: 42 patients with moderate-to-very-severe COPD (28 men, 14 women) underwent symptom-limited incremental cardiopulmonary exercise testing and also right-heart catheterization at rest. Abnormal pulmonary artery pressure was defined as mPAP>20 mmHg at rest. RESULTS: Resting mPAP was elevated in 32 patients (PH, mPAP=26.8+/-5.9 mmHg) and normal in 10 non-hypertensive (NPH) patients (NPH, mPAP=16.8+/-2 mmHg). There were no significant differences in lung function between the PH and NPH groups. Maximum oxygen uptake during exercise (VO2max) was significantly lower in PH (785+/-244 ml/min) than in NPH (1052+/-207 ml/min, P=0.004). Dead-space ventilation (Vd/Vt) was greater in PH (P=0.05) with higher VE/VCO2 (ratio of minute ventilation to carbon dioxide output=47.3+/-10 vs 38.6+/-3.5, P=0.025) and significantly higher arterial-end-tidal pCO2 difference [p(a-ET)CO2]. Pulmonary vascular resistance measured at rest correlated significantly with VO2max, VE/VCO2 and p(a-ET)CO2. CONCLUSIONS: In patients with COPD, abnormal pulmonary artery pressure impairs gas exchange, decreases maximum oxygen uptake during exercise and impairs exercise capacity. PMID- 19122988 TI - Indications for caesarean delivery between 1955 and 2005. AB - INTRODUCTION: We wished to evaluate the changes in the indications and rates of caesarean delivery over the past 50 years (1955-2005) at the Ljubljana Maternity Hospital, the largest maternity hospital in Slovenia and a tertiary center. METHODS: We retrospectively analyzed data obtained from delivery records, archived at the Department of Obstetrics and Gynecology, University Medical Center Ljubljana, for the selected years 1955, 1965, 1975, 1985, 1995 and 2005. The records were archived in bound books (1955), folders (1965 and 1975), on microfilm (1985) and in the national perinatal information system (1995 and 2005). Caesarean delivery rates were calculated and indications statistically analyzed. RESULTS: Of the 38.499 reviewed delivery records, 3122 were the records of caesarean deliveries. The caesarean delivery rate was 2.5% in 1955, 4.95% in 1965, 7.36% in 1975, 7.54% in 1985, 11.76% in 1995 and 15.74% in 2005. Overall, 80 different indications were registered and analyzed, 35 with an incidence>1.0%. The most frequently used indications were caesarean after caesarean, cephalopelvic disproportion, acute fetal distress and malpresentation. DISCUSSION: During the 50-year period, the parameters contained in delivery records changed as the result of new approaches and changing attitudes toward the parturient woman, the course of labor and delivery. In 1955 and 1965, maternal indications prevailed, whereas in 1995 and 2005 some new and nonobstetrical indications were introduced and the fetal and maternal indications became equally important. A general observation was that combinations of indications had a constantly high share over the analyzed period, implying that the decision on caesarean delivery was often based on several different indications. The caesarean delivery rate is increasing, therefore obstetricians should, in line with a good clinical practice, base their decisions on well founded arguments. PMID- 19122989 TI - Iatrogenic metastasis of a benign meningioma to the periosteum at the site of previous craniotomy: a case report. AB - Metastasis formation after resection of meningiomas is a rare event, predominantly occurring with malignant phenotypes. As far as we know, the presented case is the first report in the literature of iatrogenic seeding of a benign meningioma to the scalp following surgery. A 37-year-old woman was admitted because of a relapsing meningioma in the frontal lobe. In 1997, she had undergone complete excision of an atypical meningioma in same location. At follow up, three new masses were found: a bifrontal meningioma on the edge of the falx, a smaller one in the falx just under the saggital sinus and a small mass, believed to be ectopic, in the periosteum at the site of the previous craniotomy. Surgical therapy was indicated. Histologically, the ectopic tumor was an atypical meningioma, similar to the one excised 10 years previously, with no relation to the other two intracranial masses. Because of the histological similarity and the location in the old craniotomy, the ectopic tumor was believed to have developed from an implantation metastasis as a consequence of the first surgery. The authors suggest that strict adherence to oncological principles should be applied in the case of benign neoplasms in order to prevent contamination of wounds with tumor cells and potential recurrence. PMID- 19122992 TI - Pharmacological analysis of feeding in a caterpillar: different transduction pathways for umami and saccharin? AB - Neonate larvae of codling moth, Cydia pomonella (L.), modify their behavior in the presence of saccharin, monosodium glutamate (MSG), or L(+)-2-amino-4 phosphonobutyric acid (L-AP4) by commencing their feeding earlier. Previously published pharmacological analysis demonstrated that phagostimulatory effects of MSG and L-AP4 (which elicit umami taste sensation in humans) are reversed by adenylate cyclase activator and phosphodiesterase inhibitor. In this study, by measuring the time needed to start ingestion of foliage treated with mixtures of phagostimulants and signal transduction modulators, we show that phagostimulatory effects of L-aspartate (the third hallmark umami substance) are also abolished by both adenylate cyclase activator and phosphodiesterase inhibitor, but not by phospholipase C inhibitor. However, stimulatory effects of hemicalcium saccharin were affected only by phospholipase C inhibitor. The results suggest that codling moth neonates use different transduction pathways for perception of hemicalcium saccharin and umami. PMID- 19122993 TI - Anatomic and acoustic sexual dimorphism in the sound emission system of Phoenicoprocta capistrata (Lepidoptera: Arctiidae). AB - Both sexes of Phoenicoprocta capistrata have functional tymbals. The scanning electron microscopy revealed differences in the morphology of these organs in males and females. Male tymbals have a well-developed striated band, constituted by 21 +/- 2 regularly arranged striae whereas female tymbals lack a striated band. This type of sexual dimorphism is rare in Arctiidae. The recording of the sound produced by moths held by the wings revealed that while males produced trains of pulses organized in modulation cycles, females produced clicks at low repetition rate following very irregular patterns. Statistically, there are differences between sexes in terms of the duration of pulses, which were 355 +/- 24 micros in the case of males and 289 +/- 29 micros for females. The spectral characteristics of the pulses also show sexual dimorphism. Male pulses are more tuned (Q(10) = 5.2 +/- 0.5) than female pulses (Q (10) = 2.7 +/- 0.5) and have a higher best frequency (42 +/- 1 kHz vs. 29 +/- 2 kHz). To our knowledge, this is the first report on an arctiid moth showing sexual dimorphism in tymbal's anatomy that leads to a best frequency dimorphism. Males produce sound at mating attempts. The sounds recorded during mating are modulation cycles with the same spectral characteristics as those recorded when males are held by the wings. The morphological and acoustic features of female tymbals could indicate a process of degeneration and adaptation to conditions under which the emission of complex patterns is not necessary. PMID- 19122994 TI - Covariation between eumelanic pigmentation and body mass only under specific conditions. AB - Identifying the factors that mediate covariation between an ornament and other phenotypic attributes is important to determine the signaling function of ornaments. Sign and magnitude of a covariation may vary across environments if the expression of the ornament or of its linked genes regulating correlated phenotypes is condition-dependent. I investigated in the barn owl Tyto alba whether sign and magnitude of covariation between body mass and two heritable melanin-based plumage ornaments change with food supply, along the reproductive cycle and from the morning to the evening. Using a dataset of 1,848 measurements of body mass in 336 breeding females, I found that females displaying large black spots were heavier than conspecifics with smaller spots in the afternoon (i.e., a long time after the last feeding) but not in the morning (i.e., a short time after the last feeding). This is consistent with the recently proposed hypothesis that eumelanin-based ornaments are associated with the ability to maintain energy balance between food intake and energy expenditure. Thus, covariation between melanin-based coloration and body mass can be detected only under specific conditions potentially explaining why it has been reported in only ten out of 28 vertebrate species. The proposition that ornamented individuals achieve a higher fitness than drab conspecifics only in specific environments should be tested for other ornaments. PMID- 19122995 TI - Double mutation of the PDC1 and ADH1 genes improves lactate production in the yeast Saccharomyces cerevisiae expressing the bovine lactate dehydrogenase gene. AB - Expression of a heterologous L: -lactate dehydrogenase (L: -ldh) gene enables production of optically pure L: -lactate by yeast Saccharomyces cerevisiae. However, the lactate yields with engineered yeasts are lower than those in the case of lactic acid bacteria because there is a strong tendency for ethanol to be competitively produced from pyruvate. To decrease the ethanol production and increase the lactate yield, inactivation of the genes that are involved in ethanol production from pyruvate is necessary. We conducted double disruption of the pyruvate decarboxylase 1 (PDC1) and alcohol dehydrogenase 1 (ADH1) genes in a S. cerevisiae strain by replacing them with the bovine L: -ldh gene. The lactate yield was increased in the pdc1/adh1 double mutant compared with that in the single pdc1 mutant. The specific growth rate of the double mutant was decreased on glucose but not affected on ethanol or acetate compared with in the control strain. The aeration rate had a strong influence on the production rate and yield of lactate in this strain. The highest lactate yield of 0.75 g lactate produced per gram of glucose consumed was achieved at a lower aeration rate. PMID- 19122996 TI - Net effect of wort osmotic pressure on fermentation course, yeast vitality, beer flavor, and haze. AB - The net effect of increased wort osmolarity on fermentation time, bottom yeast vitality and sedimentation, beer flavor compounds, and haze was determined in fermentations with 12 degrees all-malt wort supplemented with sorbitol to reach osmolarity equal to 16 degrees and 20 degrees. Three pitchings were performed in 12 degrees/12 degrees/12 degrees, 16 degrees/16 degrees/12 degrees, and 20 degrees/20 degrees/12 degrees worts. Fermentations in 16 degrees and 20 degrees worts decreased yeast vitality measured as acidification power (AP) by a maximum of 10%, lowered yeast proliferation, and increased fermentation time. Repitching aggravated these effects. The 3rd "back to normal" pitching into 12 degrees wort restored the yeast AP and reproductive abilities while the extended fermentation time remained. Yeast sedimentation in 16 degrees and 20 degrees worts was delayed but increased about two times at fermentation end relative to that in 12 degrees wort. Third "back-to-normal" pitching abolished the delay in sedimentation and reduced its extent, which became nearly equal in all variants. Beer brewed at increased osmolarity was characterized by increased levels of diacetyl and pentanedione and lower levels of dimethylsulfide and acetaldehyde. Esters and higher alcohols displayed small variations irrespective of wort osmolarity or repitching. Increased wort osmolarity had no appreciable effect on the haze of green beer and accelerated beer clarification during maturation. In all variants, chill haze increased with repitching. PMID- 19122997 TI - Inulinase-expressing microorganisms and applications of inulinases. AB - In this review article, inulinase-expressing microorganisms and its potential applications in transformation of inulin into very-high-fructose syrup, bioethanol, and inulooligosaccharides are overviewed. In the past 10 years, many new inulinase producers have been obtained and many genes encoding inulinases from different microorganisms have been cloned and characterized. Some novel processes for exoinulinase overproduction have been developed for bioethanol production and ultra-high-fructose syrup. The endoinulinases have also been used for production of inulooligosaccharides from inulin and inulin-containing materials. PMID- 19122998 TI - Simultaneous nutrients and carbon removal during pretreated swine slurry degradation in a tubular biofilm photobioreactor. AB - The biodegradation potential of an innovative enclosed tubular biofilm photobioreactor inoculated with a Chlorella sorokiniana strain and an acclimated activated sludge consortium was evaluated under continuous illumination and increasing pretreated (centrifuged) swine slurry loading rates. This photobioreactor configuration provided simultaneous and efficient carbon, nitrogen, and phosphorous treatment in a single-stage process at sustained nitrogen and phosphorous removals efficiencies ranging from 94% to 100% and 70 90%, respectively. Maximum total organic carbon (TOC), NH(4) (+), and PO(4) (3-) removal rates of 80 +/- 5 g C m(r) (-3) day(-1), 89 +/- 5 g N m(r) (-3) day(-1), and 13 +/- 3 g P m(r) (-3) day(-1), respectively, were recorded at the highest swine slurry loadings (TOC of 1,247 +/- 62 mg L(-1), N-NH(4) (+) of 656 +/- 37 mg L(-1), P-PO(4) (3+) of 117 +/- 19 mg L(-1), and 7 days of hydraulic retention time). The unusual substrates diffusional pathways established within the phototrophic biofilm (photosynthetic O(2) and TOC/NH(4) (+) diffusing from opposite sides of the biofilm) allowed both the occurrence of a simultaneous denitrification/nitrification process at the highest swine slurry loading rate and the protection of microalgae from any potential inhibitory effect mediated by the combination of high pH and high NH(3) concentrations. In addition, this biofilm-based photobioreactor supported efficient biomass retention (>92% of the biomass generated during the pretreated swine slurry biodegradation). PMID- 19122999 TI - Microbial production of 2,3-butanediol from Jerusalem artichoke tubers by Klebsiella pneumoniae. AB - 2,3-Butanediol is one of the promising bulk chemicals with wide applications. Its fermentative production has attracted great interest due to the high end concentration. However, large-scale production of 2,3-butanediol requires low cost substrate and efficient fermentation process. In the present study, 2,3 butanediol production by Klebsiella pneumoniae from Jerusalem artichoke tubers was successfully performed, and various technologies, including separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF), were investigated. The concentration of target products reached 81.59 and 91.63 g/l, respectively after 40 h in batch and fed-batch SSF processes. Comparing with fed-batch SHF, the fed-batch SSF provided 30.3% higher concentration and 83.2% higher productivity of target products. The results showed that Jerusalem artichoke tuber is a favorable substrate for 2,3-butanediol production, and the application of fed-batch SSF for its conversion can result in a more cost-effective process. PMID- 19123000 TI - Characterization of extracellular polymeric substances from denitrifying organism Comamonas denitrificans. AB - Extracellular polymeric substances (EPS) play an important role in the formation and activity of biofilms in wastewater treatment (WWT). The EPS of the denitrifying biomarker Comamonas denitrificans strain 110, produced in different culture media and growth modes, were characterized. The EPS mainly contained protein (3-37%), nucleic acids (9-50%), and carbohydrates (3-21%). The extracellular DNA was found to be important for initial biofilm formation since biofilm, but not planktonic growth, was inhibited in the presence of DNase. The polysaccharide fraction appeared to consist of at least two distinct polymers, one branched fraction (A) made up of glucose and mannose with a molecular weight around 100 kDa. The other fraction (B) was larger and consisted of ribose, mannose, glucose, rhamnose, arabinose, galactose, and N-acetylglucosamine. Fraction B polysaccharides were mainly found in capsular EPS which was the dominant type in biofilms and agar-grown colonies. Fraction A was abundant in the released EPS, the dominant type in planktonic cultures. Biofilm and agar-grown EPS displayed similar overall properties while planktonic EPS showed clear compositional disparity. This study presents results on the physiology of a key WWT organism, which may be useful in the future development of improved biofilm techniques for WWT purposes. PMID- 19123001 TI - Enhancement of display efficiency in yeast display system by vector engineering and gene disruption. AB - Vector engineering and gene disruption in host cells were attempted for the enhancement of alpha-agglutinin-based display of proteins on the cell surface in yeast. To evaluate the display efficiency by flow cytometric analysis, DsRed monomer fused with FLAG-tag was displayed and immunostained as a model protein. The use of leu2-d in the expression vector resulted in the enhanced efficiency and ratio of the accessible display of proteins. Moreover, the amount of displayed proteins in SED1-disrupted cells increased particularly during the stationary growth phase. The combination of these improvements resulted in the quantitatively enhanced accessible display of DsRed-monomer on the yeast cell surface. The improved yeast display system would be useful in a wider range of its applications in biotechnology. PMID- 19123002 TI - The comparison of weekly and three-weekly cisplatin chemotherapy concurrent with radiotherapy in patients with previously untreated inoperable non-metastatic squamous cell carcinoma of the head and neck. AB - PURPOSE: Several studies have shown that the concurrent administration of chemotherapy (CHT) and radiotherapy (RT) is superior to RT alone in patients with inoperable non-metastatic squamous cell carcinoma of the head and neck (InSCCHN). We compared the efficacy and safety profile of RT and concurrent cisplatin CHT given in two different schedules to patients with previously untreated InSCCHN. METHODS: Fifty patients with previously untreated InSCCHN admitted to our oncology department were included in the study. Thirty of 50 (60%) patients with a younger age or good performance status (PS) (ECOG 0-1) received cisplatin 100 mg/m(2) on a 21-day schedule (group A). Other 20 (40%) patients with older age or poor PS (ECOG 2) received cisplatin 40 mg/m(2) on a 7-day schedule (group B). Each of the 50 patients received concurrent conventional dose RT according to primer tumor location. RESULTS: The median follow-up is 12 months for group A and 12.5 months for group B. Twenty-eight (93.3%) patients in group A and 18 (90%) in group B were evaluable for response. The complete response rate was 50% in group A and 40% in group B (P > 0.05). The objective response rate was 92% in group A and 90% in group B (P > 0.05). All grade 3-4 toxic events were seen in 16 (53.3%) of group A patients and 8 (40%) of group B patients (P > 0.05). CONCLUSIONS: Comparison between two treatment modalities appears to result in statistically similar response rates and adverse event profile. A randomized phase III trial is required to confirm the safety and efficacy of weekly cisplatin therapy in patients with poor PS and/or older age at diagnosis. PMID- 19123003 TI - Expression of ERCC1 and class III beta-tubulin in non-small cell lung cancer patients treated with a combination of cisplatin/docetaxel and concurrent thoracic irradiation. AB - INTRODUCTION: The expression of excision repair cross-complementation group 1 (ERCC1) is reported to be correlated with resistance to platinum-based drugs. Class III beta-tubulin is reported to be correlated with resistance to taxanes. METHODS: In the present study, we evaluated whether ERCC1 and class III beta tubulin expression could be used to predict progression-free and/or overall survival in 34 patients with locally advanced non-small cell lung cancer (NSCLC) receiving concurrent chemoradiation therapy with cisplatin and docetaxel, and immunohistochemistry was used to examine the expression of these two proteins in tumor samples obtained from the patients. RESULTS: Immunostaining for ERCC1 and class III beta-tubulin was positive in 16 and 12 patients, respectively. A significant correlation was observed between ERCC1 expression and response to chemotherapy (P = 0.012), and between class III beta-tubulin expression and histology (P = 0.029). Patients negative for ERCC1 had a significantly longer median progression-free (62.5 vs. 36 weeks, P = 0.009), but not overall (171 vs. 50.5 weeks, P = 0.208), survival than those positive for ERCC1. Expression of class III beta-tubulin was not correlated with progression-free or overall survival (P = 0.563 and P = 0.265, respectively). Multivariate analysis adjusting for possible confounding factors showed that negative ERCC1 expression (hazard ratio = 3.972, P = 0.009) was a significantly favorable factor for progression free survival. CONCLUSIONS: This retrospective study indicates that immunostaining for ERCC1 may be useful for predicting survival in NSCLC patients receiving concurrent chemoradiotherapy with cisplatin and docetaxel, and can provide information critical for planning personalized chemotherapy. PMID- 19123004 TI - Overexpressed glutamine synthetase gene modifies nitrogen metabolism and abiotic stress responses in rice. AB - Glutamine synthetase (GS; EC 6.3.1.2) is a key enzyme in nitrogen metabolism; it catalyzes the critical incorporation of inorganic ammonium into glutamine. Two full-length cDNAs that encode the rice (Oryza sativa) cytosolic glutamine synthetase1 genes (OsGS1;1 and OsGS1;2) were isolated from a Minghui 63 normalized cDNA library, and glnA encoding GS in Escherichia coli was isolated by PCR amplification. Transformants for GS gene (GS1;1, GS1;2, and glnA) in rice were produced by an Agrobacterium tumefaciens-mediated transformation method, and transcripts of GS gene accumulated at higher levels in the primary transgenic plants. Our results indicated an increased metabolic level in GS-overexpressed plants, which showed higher total GS activities and soluble protein concentrations in leaves and higher total amino acids and total nitrogen content in the whole plant. Decreases in both grain yield production and total amino acids were observed in seeds of GS-overexpressed plants compared with wild-type plants. In addition, GS1;2-overexpressed plants exhibited resistance to Basta selection and higher sensitivity to salt, drought, and cold stress conditions, whereas the other two types of GS-overexpressed plants failed to show any significant changes for these stress conditions compared with wild-type plants. PMID- 19123005 TI - Innate and acquired immunity in psoriasis. PMID- 19123006 TI - 6-Benzylaminopurine stimulates melanogenesis via cAMP-independent activation of protein kinase A. AB - Melanogenesis is a physiological process that results in the synthesis of melanin pigments, which play a crucial protective role against skin photocarcinogenesis. The present study was designed to determine the effects of 6-benzylaminopurine (6 BAP) on melanogenesis and elucidate the molecular events of melanogenesis induced by 6-BAP. To elucidate the pigmenting effect of 6-BAP and its mechanism, several experiments were performed in B16 melanoma cells. Melanin content, tyrosinase activity, cAMP production, and Western blots for proteins which are involved in melanogenesis were introduced in this study. Melanin content and tyrosinase activity increased in response to treatment with 6-BAP in a concentration dependent manner. The tyrosinase, TRP-1, TRP-2 and MITF protein levels were found to increase significantly in response to 6-BAP in a time-dependent manner. In addition, Western blot analysis revealed that 6-BAP increased the phosphorylated level of CRE-binding protein. The increased melanin synthesis that was induced by treatment with 6-BAP treatment was reduced significantly in response to co treatment with H-89 [a protein kinase A (PKA) inhibitor], whereas co-treatment with SB203580 (a p38 MAPK inhibitor) and Ro-32-0432 (a PKC inhibitor) did not attenuate the increase in melanin content levels that was induced by 6-BAP. In a cAMP production assay, 6-BAP did not increase the intracellular cAMP level. These findings suggest that 6-BAP activates PKA via a cAMP-independent pathway and subsequently stimulates melanogenesis by up-regulating MITF and tyrosinase expression. PMID- 19123007 TI - Polycystic ovary syndrome and fibrocystic breast disease: is there any association? AB - OBJECTIVE: We aimed to investigate the association between a polycystic ovary syndrome (PCOS) and fibrocystic breast disease. METHODS: A total of 93 women, aged between 17 and 36 years, not using oral contraceptives, were entered in this case-control study. Laboratory, clinical and ultrasound findings were used to diagnose PCOS. The study group was consisted of 53 PCOS women and the control group consisted of 40 women. Breast ultrasonography was performed for all patients. Fibrocystic breast disease is described as common benign changes involving the tissues of the breast. RESULTS: Twenty-one (39.6%) of 53 women with a PCOS had fibrocystic breast disease. Five (8%) of 40 controls had fibrocystic breast disease. The difference between the groups was statistically significant (p = 0.004). Relative risk (95% CIs) was 3.17 (1.31-7.68). Overall sonographic benign breast pathologies were significantly higher in the PCOS group (p = 0.036). CONCLUSION: This study showed a statistically significant association between a PCOS and fibrocystic breast disease. Women with a PCOS should be evaluated for fibrocystic breast disease. PMID- 19123008 TI - Phenotypic and genotypic diversity among strains of Aureobasidium pullulans in comparison with related species. AB - Intra-specific diversity of 200 Aureobasidium pullulans strains isolated from different sources and their relatives Kabatiella lini CBS 125.21 T and Hormonema prunorum CBS 933.72 T were studied by assessment of macromorphological, and physiological tests, sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique (SDS-PAGE) of whole-cell proteins as well as enterobacterial repetitive intergenic consensus (ERIC)-, repetitive extragenic palindromic (REP)- and BOX PCR techniques (collectively known as rep-PCR). Rep-PCR is an efficient procedure for discrimination of A. pullulans in terms of simplicity and rapidity. RFLP-PCR technique was applied for the identification of A. pullulans isolates and distinction from related species. This technique was insufficient for investigation of intra-specific diversity. The tested strains of A. pullulans could be divided into two groups based on their macromorphological, protein patterns obtained after SDS-PAGE as well as rep-PCR patterns. The first group of strains shared similar characteristics and was very different from the second one, designated as "complex group", consisting of strains with very little similarities within the group. Phenetic analysis of ERIC banding patterns failed to group the isolates on the basis of their substrate or geographical origin. Using 18S rDNA gene sequence analysis of selected isolates, three strains: HoHe3 km, A. pullulans DSM 62074 and H. prunorum CBS 933.72 T were distinguished from all other analysed members of genera Aureobasidium and Kabatiella. PMID- 19123009 TI - Evaluation of the efficacy of strains of Steinernema carpocapsae Santa Rosa and ALL (Steinernematidae: Rhabditida) to control engorged female Anocentor nitens (Acari: Ixodidae). AB - In view of the need to combat the generalized spread of resistance in ticks to commercial acaricides, the objective of this study was to evaluate the action of entomopathogenic nematodes (Steinernema carpocapsae, strains Santa Rosa and ALL) on engorged female Anocentor nitens. Five ticks per Petri dish were exposed to concentrations of 500, 5,000, or 25,000 infective juveniles of S. carpocapsae for 72 h. After transferring the ticks to clean plates, biological parameters were analyzed. Related to strains Santa Rosa, the period of pre-oviposition (p = 0.0001), oviposition (p = 0.041), and the mass weight of eggs (p = 0.005) showed significant differences between the control group and treated group. When the strain ALL was tested, the control and treated groups differed between the periods of pre-oviposition (p = 0.001), oviposition (p = 0.001), and egg mass weight (p = 0.01). The egg mass conversion was less significant in the groups when exposed to strains Santa Rosa (p = 0.002) and ALL (p = 0.001) relative to the control. The efficacy of both entomopathogenic nematode strains used in this study was comparable to other biological control agents, showing their potential against A. nitens in the laboratory. PMID- 19123011 TI - Response to the expertise: procedures and methods of benefit assessments for medicines in Germany, by Geertruida E. Bekkering and Jos Kleijnen. PMID- 19123010 TI - Characterization of autonomous Dart1 transposons belonging to the hAT superfamily in rice. AB - An endogenous 0.6-kb rice DNA transposon, nDart1-0, was found as an active nonautonomous element in a mutable virescent line, pyl-v, displaying leaf variegations. Here, we demonstrated that the active autonomous element aDart in pyl-v corresponds to Dart1-27 on chromosome 6 in Nipponbare, which carries no active aDart elements, and that aDart and Dart1-27 are identical in their sequences and chromosomal locations, indicating that Dart1-27 is epigenetically silenced in Nipponbare. The identification of aDart in pyl-v was first performed by map-based cloning and by detection of the accumulated transposase transcripts. Subsequently, various transposition activities of the cloned Dart1-27 element from Nipponbare were demonstrated in Arabidopsis. Dart1-27 in Arabidopsis was able to excise nDart1-0 and Dart1-27 from cloned sites, generating footprints, and to integrate into new sites, generating 8-bp target site duplications. In addition to Dart1-27, Nipponbare contains 37 putative autonomous Dart1 elements because their putative transposase genes carry no apparent nonsense or frameshift mutations. Of these, at least four elements were shown to become active aDart elements in transgenic Arabidopsis plants, even though considerable sequence divergence arose among their transposases. Thus, these four Dart1 elements and Dart1-27 in Nipponbare must be potential autonomous elements silenced epigenetically. The regulatory and evolutionary implications of the autonomous Dart1 elements and the development of an efficient transposon-tagging system in rice are discussed. PMID- 19123013 TI - Symbiobacterium lost carbonic anhydrase in the course of evolution. AB - Recent genetic studies have elucidated that carbonic anhydrase (CA; EC 4.2.1.1), a ubiquitous enzyme catalyzing interconversion between CO(2) and bicarbonate, is essential for microbial growth under ambient air but not under high-CO(2) air. The irregular distribution of the phylogenetically distinct types of CA in the prokaryotic genome suggests its complex evolutionary history in prokaryotes. This paper deals with the genetic defect of CA in Symbiobacterium thermophilum, a syntrophic bacterium that effectively grows on CO(2) generated by other bacteria. Phylogenetic analysis based on 31 ribosomal protein sequences demonstrated the affiliation of Symbiobacterium with the class Clostridia with 100% bootstrap support. The phylogeny of beta- and gamma-type CA distributed among Clostridia supported the view that S. thermophilum and several related organisms lost this enzyme during the course of evolution. The loss of CA could be based on the availability of a high level of CO(2) in their living environments. PMID- 19123012 TI - Origin of axial prestretch and residual stress in arteries. AB - The structural protein elastin endows large arteries with unique biological functionality and mechanical integrity, hence its disorganization, fragmentation, or degradation can have important consequences on the progression and treatment of vascular diseases. There is, therefore, a need in arterial mechanics to move from materially uniform, phenomenological, constitutive relations for the wall to those that account for separate contributions of the primary structural constituents: elastin, fibrillar collagens, smooth muscle, and amorphous matrix. In this paper, we employ a recently proposed constrained mixture model of the arterial wall and show that prestretched elastin contributes significantly to both the retraction of arteries that is observed upon transection and the opening angle that follows the introduction of a radial cut in an unloaded segment. We also show that the transmural distributions of elastin and collagen, compressive stiffness of collagen, and smooth muscle tone play complementary roles. Axial prestresses and residual stresses in arteries contribute to the homeostatic state of stress in vivo as well as adaptations to perturbed loads, disease, or injury. Understanding better the development of and changes in wall stress due to individual extracellular matrix constituents thus promises to provide considerable clinically important insight into arterial health and disease. PMID- 19123014 TI - Internal repetition and intraindividual variation in the rDNA ITS1 of the anopheles punctulatus group (Diptera: Culicidae): multiple units and rates of turnover. AB - The rapid divergence of repetitive sequences makes them desirable markers for phylogenetic studies of closely related groups, provided that a high level of sequence homogeneity has been maintained within species. Intraspecific polymorphisms are found in an increasing number of studies now, and this highlights the need to determine why these occur. In this study we examined intraindividual variation present in the first ribosomal internal transcribed spacer (ITS1) from a group of cryptic mosquito species. Individuals of the Anopheles punctulatus group contained multiple ITS1 length variants that ranged from 1.2 to 8.0 kb. Nucleotide and copy number variation for several homologous internal repeats is common, yet the intraspecific sequence divergence of cloned PCR isolates is comparable to that of other mosquito species (~0.2-1.5%). Most of the length variation is comprised of a 5'-ITS1 repeat that was identified as a duplication of a conserved ITS2 region. Secondary structure conservation for this repeat is pronounced and several repeat types that are highly homogenized have formed. Significant interspecific divergence indicates a high rate of evolutionary change for this spacer. A maximum likelihood tree constructed here was congruent with previous phylogenetic hypotheses and suggests that concerted evolution is also accompanied by interpopulation divergence. The lack of interindividual differences and the presence of homogenized internal repeats suggest that a high rate of turnover has reduced the overall level of variation. However, the intraindividual variation also appears to be maintained by the absence of a single turnover rate and the complex dynamics of ongoing recombination within the spacer. PMID- 19123015 TI - A child with eosinophilia, Loeffler endocarditis, and acute lymphoblastic leukemia. AB - We present an 8-year-old male with Loeffler endocarditis and acute lymphoblastic leukemia with hypereosinophilia (ALL/Eo) who initially presented with a 3-month history of peripheral eosinophilia thought to be due to visceral larval migrans. Despite treatment for Toxocara, his leukocytosis persisted and he developed mitral valve insufficiency and congestive heart failure. Myocardial biopsy revealed fibrosis and thrombus formation indicative of Loeffler endocarditis, and a peripheral smear showed pre-B-cell acute lymphoid leukemia. This unique case highlights a rare, yet serious sequella of prolonged eosinophilia. PMID- 19123016 TI - Midterm results of the Ross procedure in a pediatric population: bicuspid aortic valve is not a contraindication. AB - Our objective was to analyze retrospectively the short- and midterm results of the Ross operation in children and the impact of bicuspid aortic valve (BAV) disease on outcome. From 1991 to 2003, 41 patients (26 male, 15 female) underwent a Ross procedure. Aortic disease was congenital in all but one. Sixty-six percent had BAV. Mean age at operation was 10.13 +/- 5.6 years (range, 0.4-18.3 years). Root replacement technique was performed in all but two (inclusion technique). There were two early deaths. Mean follow-up was 6 +/- 3.8 years (range, 0.1-14 years). The autograft (neo-aorta) presented absent, trivial, mild, and moderate regurgitation in 42%, 46%, 10%, and 2%, respectively, at latest follow-up. Root dilation was seen in 64% of the patients (mean Z-score, +3.53 +/- 0.04). Four patients (9%) required allograft replacement, two for endocarditis and two for stenosis. Allograft stenosis (gradient >20 mmHg) was detected in 44% of the remaining patients, without symptoms or the need for reintervention. Estimated freedom from allograft replacement at 5 and 10 years was 97% and 89%. Left ventricular dimensions and function were normal in all patients. No difference was found between patients with BAV and those with tricuspid aortic valves in aortic regurgitation or root dilation. BAV was not identified as a risk factor for root dilation. In conclusion, the Ross operation remains an excellent option for aortic valve replacement in children. BAV is not associated with a worse outcome and is not a risk factor for aortic root dilation at medium-term follow up. Long-term follow-up is, however, required. PMID- 19123019 TI - Periareolar breast reduction. AB - A personal approach to periareolar breast reduction is presented. The circular demarcation of periareolar skin must be limited to twice the demarcated areolar diameter (2 x 4 = 8 cm). A cylindrical resection of volume is removed, as in Strombeck's technique, from below the areola to the aponeurosis of the pectoral muscle. Through this space, two to four "keel-like" parenchymal pieces are removed to reduce the breast at the cardinal points. The breast cone is assembled by suturing the raw areas from bottom to top, hyperprojecting it. A "round-block" suture around the areola is made, and the procedure is concluded with skin suture. Indications, limitations, and possibilities for use of the technique are analyzed, and the equation is summarized as follows: residual ptosis versus long scars inside the limits and indications of more or less 300 g of removed volume and small or medium breast ptosis. Only after the first 10 cases was the limit of the technique determined. Therefore, these results were not homogeneous. The results were satisfactory and regular for 68 of the 78 patients subjected to surgery and follow-up evaluation. PMID- 19123018 TI - "Coagulum suction": a simple trick for the reduction of postoperative hematoma associated with facelift. AB - BACKGROUND: Facelift is currently one of the most requested procedures among consumers of aesthetic plastic surgery. Like any operation, it is accompanied by a variety of potential complications, with postoperative bleeding probably the most frequent. Hematomas can cause hyperpigmentation,contour changes due to subcutaneous scarring, prolongation of healing, and necrosis of the skin flap. The most common treatment is manual expression of the blood coagula. Needle aspiration sometimes cannot be used because of the viscosity of the coagula. METHODS: Seven patients underwent a new method of hematoma removal from wounds. One to two sutures were removed above the hairline under local anesthesia and a short liposuction cannula, 2.5 mm in diameter and 10-15 cm in length with one or two openings, was used for coagulum suction. RESULTS: All treated patients had the hematoma suction procedure without complications and with satisfactory outcomes and no subsequent bleeding. CONCLUSION: Coagulum suction is a simple and fast method that can shorten the recovery following a facelift.This method cannot replace surgical revision in cases in which there is substantial and/or persistent bleeding.However, it can reduce hematomas that are large enough to delay healing and interfere with convalescence, yet too small to warrant surgical revision. PMID- 19123020 TI - Body contouring with solid silicone implants. AB - BACKGROUND: Body-contouring implants are becoming increasingly popular and more accepted and requested for aesthetic purposes. These implants must be placed in a safe and reproducible plane of anatomic dissection for a successful long-term outcome. Poor implant placement techniques result in greater complications such as asymmetry, implant migration, capsular contracture, and infection. This article discusses (1) the history and the evolution of body-contouring implants, (2) the different techniques and dissection planes in which to put the implants in the calf, gluteal, pectoral, and triceps/biceps areas, and (3) the complications associated with the different implantation techniques. METHODS: The different anatomical areas that have been treated with solid silicone implants are the gluteus, pectoral, calf, deltoids, biceps, triceps, and trapezium. RESULTS: Determining the ideal plane for implant placement is the most important surgical goal for satisfactory aesthetic results. CONCLUSION: Aesthetic contouring of the chest, back, arms, buttocks, and calf can be done safely with solid silicone implants. Proper pocket dissection and location are paramount to successful implant placement and decreased complications. PMID- 19123021 TI - Treatment of axillary hyperhidrosis/bromidrosis using VASER ultrasound. AB - BACKGROUND: Current methods of treatment for axillary hyperhidrosis and/or bromidrosis are palliative (use of topical aluminum chloride or injections of botulinum toxin type A) or surgically based for more permanence (excisional surgery, endoscopic transthoracic sympathectomy, liposuction/curettage). The surgical approaches have mixed effectiveness and incur the risk of significant side effects and complications. METHODS: Thirteen patients (3 males, 10 females) with significant axillary hyperhidrosis and/or bromidrosis were recruited, treated with the VASER ultrasound, and followed for 6 months. Preoperative assessment of the impact of hyperhidrosis and/or bromidrosis on lifestyle and the degree of sweat/odor were completed. Postoperative assessment of changes relative to lifestyle and degree of sweat/odor reduction and patient and surgeon satisfaction were completed. RESULTS: Eleven of 13 patients had significant reduction in sweat/odor and had no recurrence of significant symptoms at 6 months. Two patients had a reduction in sweat/odor but not to the degree desired by the patients. No significant complications were noted. A simple amplitude and time protocol was established that provides consistent and predictable therapy. The complete procedure takes less than 1 h to treat two axillae using local anesthetic. CONCLUSION: The VASER is safe and effective for treatment of axillary hyperhidrosis/bromidrosis. The method is minimally invasive with immediate return to basic activities and only temporary minor restriction of arm movement. At 6 months the treatment appears to be long-lasting, but further follow-up is required for verification of permanence. This method has become the standard of care for the treatment of axillary hyperhidrosis/bromidrosis in the authors' practice. PMID- 19123022 TI - Risk factors for nodal metastasis and recurrence among patients with papillary thyroid microcarcinoma: differences in clinical relevance between nonincidental and incidental tumors. AB - BACKGROUND: Papillary microcarcinoma (PMC) is a subgroup of papillary thyroid carcinoma (PTC) measuring 1.0 cm or less in diameter. Herein we focused on the search for risk factors predicting nodal metastasis and recurrence in PMCs, analyzing differences in presentation, treatment, and prognosis between nonincidental and incidental tumors. METHODS: From January 1998 to May 2007, 149 patients had a diagnosis of PMC in our department. A cross-sectional study of 76 patients with nonincidental and 73 patients with incidental PMC was carried out. Demographic data, diagnostic results, tumor characteristics, risk assessment, surgical treatment, and postoperative and follow up results were evaluated. RESULTS: Cytology detected thyroid cancer and nodal involvement in nonincidental PMC. Mean tumor size was significantly larger in nonincidental PMC (7.5 vs. 4.2 mm), which was commonly found within a normal thyroid gland or Hashimoto's thyroiditis, while incidental PMC was associated with a multinodular goiter. TNM staging system showed a higher cancer stage (IVA) in nonincidental. At multivariate analysis, capsular invasion and a nonincidental diagnosis were the two independent factors significantly affecting nodal metastasis. All patients with nonincidental PMC underwent iodine-131 ablation therapy after surgery compared with 49 patients with incidental. Nodal metastasis at diagnosis was the only factor influencing recurrence which was found in three nonincidental cases: two in the lateral and one in the central neck compartments. CONCLUSIONS: Several PMCs presented with risk-free clinical courses. Some nonincidental tumors had a more aggressive behavior and a tendency to recurrence. In these cases, early detection and aggressive treatment are mandatory as for conventional PTC according to risk stratification and cancer stage. PMID- 19123023 TI - Laparoscopic surgical skills assessment: can simulators replace experts? AB - INTRODUCTION: Global Rating Scales (GRS) quantify and structure subjective expert assessment of skill. Hybrid simulators measure performance during physical laparoscopic tasks through instrument motion analysis. We assessed whether motion analysis metrics were as accurate as structured expert opinion by using GRS. METHODS: A random sample of 10 consultant laparoscopic surgeons, 10 senior trainees, and 10 novice students were assessed on a Sharp Dissection task. Coded video footage was reviewed by two blinded assessors and scored using a Likert Scale. Correlation with metrics was tested using Spearman's rho. Inter-rater reliability was measured using intraclass correlation coefficient (ICC). RESULTS: Strongest GRS-Metric correlations were found for Time/Motion/Progress with Time (Spearman's rho 0.88; p < 0.05) and Instrument Handling with Path Length (Spearman's rho 0.8; p < 0.05). Smoothness correlated with Respect for Tissue in Rater 1 (rho 0.68) but not Rater 2 (rho 0.18). Mean GRS showed stronger inter rater agreement than individual scale components (ICC 0.68). Correlation coefficients with actual experience group were 0.58-0.74 for mean GRS score and 0.67-0.78 for metrics (Spearman's rho, p < 0.05). CONCLUSIONS: Metrics correlate well with GRS assessment, supporting concurrent validity. Metrics predict experience level as accurately as global rating and are construct valid. Hybrid simulators could provide resource-efficient feedback, freeing trainers to concentrate on teaching. PMID- 19123025 TI - Effectiveness of radiofrequency ablation of hepatocellular carcinoma. PMID- 19123024 TI - MicroRNA and colorectal cancer. AB - MicroRNAs are small 19 to 22 nucleotide sequences of RNA that participate in the regulation of cell differentiation, cell cycle progression, and apoptosis. MicroRNAs act much like small interfering RNA, annealing with RISC, to cleave messenger RNA, and microRNAs exert translational inhibition that is incompletely understood. They are important factors in tumorigenesis and have been the subject of research in many types of cancers, including colon cancer. MicroRNAs may be abnormally down-regulated or up-regulated in colon-cancer tissue. Artificial dysregulation of certain microRNAs will trigger tumorigenesis or apoptosis depending on which microRNA is manipulated. Although the natural mechanisms for the dysregulation of microRNAs is still largely unknown, one theory tested in colon cancers proposes that DNA hypermethylation leads to down-regulation of certain microRNAs. Specific microRNA expression patterns help characterize specific cancers and may be used as a prognostication factor and in following patient response to 5-fluorouracil chemotherapy. This article reviews the existing literature pertaining to the study of microRNA in colorectal cancer. PMID- 19123026 TI - Profile of breast cancer in a group of women in a developing country in South Asia: is there a difference? AB - BACKGROUND: Breast cancer is the commonest cancer among Sri Lankan women. The aim of this study was to document the breast cancer profile of a group of Sri Lankan women and compare it with regional data. Patient-tumor characteristics and predicted prognosis are compared with the immune profile. METHODS: A total of 814 Sri Lankan women with breast cancer were studied, with their information retrieved from patient records. Tumor type and grade were reassessed on routine tissue sections. The Nottingham Prognostic Index (NPI) was calculated. Estrogen receptors (ER) and human epidermal receptor 2 (HER2) were assessed using Dako antibodies. Strong nuclear staining for ER in >10% of tumor cells and strong, complete cell membrane staining (3+) for HER2 were regarded as positive. An SPSS 16 software program and the chi-squared test were used for statistical analysis. RESULTS: The highest prevalence (32%) was in the 50- to 59-year age cohort (mean +/- SD 51.88 +/- 11.939 years). In all, 58% of the tumors measured between 2 and 5 cm. Most (52%) were moderately differentiated and were invasive ductal carcinomas (86.3%). Regional lymph node metastasis was present in 41% of the patients. ER was expressed in 31.7% and was more frequent in women >35 years (p < 0.024). HER2 was found in 14.5% of tumors. Its expression was lower in ER positive tumors (p < 0.000). Well-differentiated tumors were frequently ER positive (p < 0.000) and HER2-negative (p < 0.001). The NPI was better for ER positive (p < 0.000) and HER2-negative tumors (p < 0.028). CONCLUSIONS: The overall profile of breast cancer and immune characteristics of Sri Lankan women in this study was largely comparable to profiles documented elsewhere in the region despite the lower prevalence of ER. PMID- 19123027 TI - Design and implementation of a web-based system for intraoperative consultation. AB - BACKGROUND: With the use of electronic information distribution and telecommunication technologies, surgical teleconsultation is possible as a vehicle for consulting with experts remotely without their physical presence in the operating room (OR). This study evaluated real-time teleconsultation from the OR to remote consultants in the Russian Research Center of Surgery, Moscow, Russia and the Fundeni Clinical Hospital, Bucharest, Romania. METHODS: We evaluated the effectiveness of teleconsultation using a secure website interface where consultants could navigate through multimedia-based electronic documentation of a surgical procedure and identify the anatomic landmarks underlying the need for consultation. Additionally, management of a remote camera view by consultants was evaluated. RESULTS: Fifteen thyroidectomies and parathyroidectomies were studied to confirm by teleconsultation the identity of 22 recurrent laryngeal nerves (RLNs). There was no Internet connectivity interruption or dropped signal, and the bandwidth was consistently greater than 1 Mbps. Consultants spent an average of 6 min to review an average of 35 min of surgical records to identify the 22 RLNs. CONCLUSIONS: This study validated a system for real-time teleconsultation using web-based surgical records. In addition, the ability of the consultant to manage the camera view remotely without interrupting the surgical procedure was confirmed. PMID- 19123028 TI - Idiopathic massive spontaneous hemothorax: adhesion disruption. AB - BACKGROUND: Hemothorax has been reported to occur along with spontaneous pneumothorax due to adhesion disruption. Rupture of pleural adhesions spontaneously or after unnoticeable trivial trauma causing massive hemothorax alone is rare. METHODS: We present a series of seven cases of idiopathic massive spontaneous hemothorax due to adhesion disruption, of which all required emergency thoracotomy with ligation or cauterization of bleeding adhesions. RESULTS: Six patients had bleeding pleural lung adhesions of which five involved the upper lobes. Another had bleeding from pleuropericardial adhesions. All patients are doing well on follow-up. CONCLUSIONS: Disruption of pleural adhesions may cause massive hemothorax, requiring early surgical intervention. After thoracotomy the outcome in these patients is excellent. PMID- 19123029 TI - C-arm cone beam computed tomographic needle path overlay for fluoroscopic-guided placement of translumbar central venous catheters. AB - C-arm cone beam computed tomography is an advanced 3D imaging technology that is currently available on state-of-the-art flat-panel-based angiography systems. The overlay of cross-sectional imaging information can now be integrated with real time fluoroscopy. This overlay technology was used to guide the placement of three percutaneous translumbar inferior vena cava catheters. PMID- 19123030 TI - Characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene from the halotolerant alga Dunaliella salina and inhibition of its expression by RNAi. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) displays important functional diversity in mammalian and plants. So far, however, studies on GAPDH have not included the halotolerant, unicellular green alga Dunaliella salina. In the present study, a GAPDH cDNA was cloned and sequenced from D. salina. It was 1394 bp long, with an open reading frame of 1128 bp encoding 376 amino acid residues, and shared a high homology with other organisms. The coding region of the gene was heterologously expressed in E. coli, confirming that the gene cloned from D. salina is indeed GAPDH. Furthermore, the recombinant plasmid p7NBTFIR was constructed to express hairpin RNA (hpRNA) containing sequences homologous to the GAPDH gene to investigate the expression profile of GAPDH by RNAi in D. salina. The results of real-time quantitative PCR revealed that the relative transcription levels of the GAPDH gene in transformants G1 and G2 were reduced to 41.2% and 67.4%, respectively, of the wild-type D. salina. Observations under phase-contrast microscopy showed that the motility of the transformants cell was sluggish. The results of a photoaccumulation experiment showed that the cell motility of transformants G1 and G2 was less active than that of wild-type D. salina. The findings of this study may be useful for further studies on the subcellular localization and functional analysis of the GAPDH gene in microalgae. PMID- 19123031 TI - Vertical transmission of Chlamydia trachomatis in Chongqing China. AB - This is the first study to investigate vertical transmission of Chlamydia trachomatis in Chongqing China. For this study, 300 cervical swab samples from pregnant women and 305 nasopharygeal swab samples from their babies (605 specimens) were collected for nest polymerase chain reaction (nPCR) of the ompl gene, which encodes the major outer membrane protein (MOMP) and typed C. trachomatis using Cleavase fragment-length polymorphism (CFLP) labeled with digoxin. From these samples, 11% (33/300) of pregnant women samples were successfully amplified. The vertical transmission rate of C. trachomatis from mother to baby was 24% (8/33). The vertical transmission rates were 66.7% (6/9) for mothers with vaginal delivery and 8.3% (2/24) for those with cesarean section. The incidence of premature membrane rupture among C. trachomatis positive pregnant women was 30.3% (10/33), which was greater than among those who were C. trachomatis-negative (13.5%, 36/267; chi(2) = 4.2; p < 0.05). Four genotypes including type E (3 pairs), type F (2 pairs), type H (2 pairs), and type D (1 pair) were observed by CFLP assay labeled with digoxin and confirmed by DNA sequencing in the 16 C. trachomatis-positive samples from eight pregnant women and their eight infants. Each pair of matched maternal-infantile samples showed identical CFLP. This study showed the incidence of C. trachomatis infection in pregnant women, the vertical transmission rate for C. trachomatis, and the genotypes of C. trachomatis in Chongqing, China. The CFLP assay labeled at the 5' end of the forward primer with digoxin was first used successfully to genotype of C. trachomatis. As a promising method for C. trachomatis genotyping, CFLP had good sensitivity, reproducibility, and simplicity and no radioactive contamination. PMID- 19123032 TI - Identification of small noncoding RNAs in Helicobacter pylori by a bioinformatics based approach. AB - Small noncoding RNAs (sRNAs) are a group of regulatory RNA molecules normally without a protein-coding function. In recent years, the importance of sRNAs as mediators of gene expression in bacteria has begun to be recognized. More than 70 sRNAs have been known in Escherichia coli. However, little is known about sRNAs in Helicobacter pylori, a human pathogen associated with gastric diseases. Here, we systematically identified sRNAs in the H. pylori genome by a computational approach based on gene location, sequence conservation, promoter and terminator search, and secondary structure. Among a total of six candidate sRNAs initially predicted, two novel sRNAs (IG-443 and IG-524) were confirmed by Northern blot and reverse transcription-polymerase chain reaction (RT-PCR). Virtually, they were a class of natural antisense transcripts, which were complementary to partial sequences of the following genes: flagellar motor switch gene (fliM) and fumarase (fumC). Taken together, the results indicate that there exist novel sRNAs in H. pylori and these RNAs might play a potential role in regulating gene expression. PMID- 19123033 TI - Transformation of L-tyrosine to L-dopa by a novel fungus, Acremonium rutilum, under submerged fermentation. AB - The present study deals with the transformation of L-tyrosine to L-dopa by Acremonium rutilum, a fungal tyrosinase producer, isolated from decomposed banana stud. This appears to be the first report on A. rutilum as a polyphenoloxidase producer with both cresolase and catecholase activity. Enriched Czapek-Dox agar was used for plate assay screening. Enriched potato dextrose broth was used for optimization studies, which induced high levels of L-dopa under submerged fermentation. A. rutilum gave the maximum L-dopa production (0.89 mg/ml) and tyrosinase activity (1095 U/mg) under the optimized parameters, that is, a temperature of 25 degrees C, pH 5.5, an inoculum size of 2.5 ml, and an incubation time of 72-120 h, with L-tyrosine (5 mg/ml) as substrate. Five resolved bands, with R(f) values of 0.73, 0.60, 0.54, 0.37, and 0.26, were observed, which confirmed the presence of L-dopa. This study involves the elevated profile of L-dopa production. Such study is needed, as L-dopa has the ability to control Parkinson's disease. PMID- 19123034 TI - Polymorphism in hybrid male sterility in wild-derived Mus musculus musculus strains on proximal chromosome 17. AB - The hybrid sterility-1 (Hst1) locus at Chr 17 causes male sterility in crosses between the house mouse subspecies Mus musculus domesticus (Mmd) and M. m. musculus (Mmm). This locus has been defined by its polymorphic variants in two laboratory strains (Mmd genome) when mated to PWD/Ph mice (Mmm genome): C57BL/10 (carrying the sterile allele) and C3H (fertile allele). The occurrence of sterile and/or fertile (wild Mmm x C57BL)F1 males is evidence that polymorphism for this trait also exists in natural populations of Mmm; however, the nature of this polymorphism remains unclear. Therefore, we derived two wild-origin Mmm strains, STUS and STUF, that produce sterile and fertile males, respectively, in crosses with C57BL mice. To determine the genetic basis underlying male fertility, the (STUS x STUF)F1 females were mated to C57BL/10 J males. About one-third of resulting hybrid males (33.8%) had a significantly smaller epididymis and testes than parental animals and lacked spermatozoa due to meiotic arrest. A further one fifth of males (20.3%) also had anomalous reproductive traits but produced some spermatozoa. The remaining fertile males (45.9%) displayed no deviation from values found in parental individuals. QTL analysis of the progeny revealed strong associations of male fitness components with the proximal end of Chr 17, and a significant effect of the central section of Chr X on testes mass. The data suggest that genetic incompatibilities associated with male sterility have evolved independently at the proximal end of Chr 17 and are polymorphic within both Mmd and Mmm genomes. PMID- 19123035 TI - Phenology of semiochemical-mediated host foraging by the western boxelder bug, Boisea rubrolineata, an aposematic seed predator. AB - The western boxelder bug (BEB), Boisea rubrolineata (Heteroptera: Rhopalidae), is a specialist herbivore of boxelder trees, Acer negundo. We tested the hypothesis that BEBs use semiochemicals to locate host trees. Headspace volatiles from trees bearing staminate inflorescences ("staminate trees") and from trees bearing pistillate inflorescences ("pistillate trees") were collected throughout the season and bioassayed in Y-tube olfactometer experiments. Headspace extracts of early-season, pollen-bearing staminate trees and midseason pistillate trees with mature samaras (seed pods) attracted female and male BEBs. By using coupled gas chromatographic-electroantennographic detection and gas chromatography-mass spectrometry, we identified and tested a five-component synthetic blend of candidate semiochemicals (hexanol, pentyl acetate, phenylacetonitrile, 2 phenethyl acetate, and trans-nerolidol). This blend attracted females, males, and fifth-instar nymphs. Phenylacetonitrile by itself was as attractive as the five component blend to both adults and nymphs. By responding to phenylacetonitrile emitted by pollen-bearing staminate trees and pistillate trees with maturing seeds, BEBs appear to track and exploit the availability of nutrient-rich food sources, suggesting that the bugs' reproductive ecology is synchronized to the phenology of their host boxelder tree. PMID- 19123036 TI - Yellow eel (Anguilla anguilla) development in NW Portuguese estuaries with different contamination levels. AB - The aims of the present study were to compare the health status of yellow eels (Anguilla anguilla) developing in three estuaries of the NW Portuguese coast with different levels of pollution and their physiological responses to combined effects of environmental variation and pollution. For this, a field study was performed using a multi-parameter approach, including eels condition indexes and biomarkers, water quality variables and other environmental factors. Sixteen biological parameters were assessed, namely: hepatosomatic index (LSI), Fulton's condition index (K), lipid peroxidation (LPO), total glutathione (TG), reduced glutathione (GSH), oxidised glutathione (GSSG), GSH/GSSG, and the activity of the enzymes acetylcholinesterase (AChE), lactate dehydrogenase (LDH), sodium potassium ATPase (Na(+)/K(+)-ATPase), ethoxyresorufin O-deethylase (EROD), glutathione S-transferases (GST), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR). Ten environmental factors were also measured in water: temperature, salinity, pH, phosphates, nitrates, nitrites, ammonium, silica, phenol and hardness. Globally, the biomarkers indicate exposure and toxic effects of pollutants on eels living in contaminated estuaries. The relationships between biological and environmental variables were assessed through redundancy analysis. K and LSI indexes, AChE and Na(+)/K(+)-ATPase, total glutathione levels and the antioxidant enzymes CAT, GR, and SOD where the factors most discriminating reference (Minho River estuary) from contaminated estuaries (Lima and Douro Rivers estuaries). Moreover, the most striking outcomes of pollutants exposure on biological responses were observed during winter, probably due to a joint effect of cold weather and pollution stress. Altogether, the results indicate that the development of eels in the polluted estuaries of Lima and Douro rivers is interfering with physiological functions determinant for their survival and performance. This may increase the mortality rates during the continental life-phase of the species and decrease the percentage of animals able to successfully complete their oceanic migration and, thus, reduce the contribution of each generation to the next one. PMID- 19123038 TI - Nonparametric modeling of the gap time in recurrent event data. AB - Recurrent event data arise in many biomedical and engineering studies when failure events can occur repeatedly over time for each study subject. In this article, we are interested in nonparametric estimation of the hazard function for gap time. A penalized likelihood model is proposed to estimate the hazard as a function of both gap time and covariate. Method for smoothing parameter selection is developed from subject-wise cross-validation. Confidence intervals for the hazard function are derived using the Bayes model of the penalized likelihood. An eigenvalue analysis establishes the asymptotic convergence rates of the relevant estimates. Empirical studies are performed to evaluate various aspects of the method. The proposed technique is demonstrated through an application to the well known bladder tumor cancer data. PMID- 19123037 TI - Up-regulation of D-serine might induce GABAergic neuronal degeneration in the cerebral cortex and hippocampus in the mouse pilocarpine model of epilepsy. AB - Epilepsy is a serious neurological disorder with neuronal loss and spontaneous recurrent seizures, but the neurochemical basis remains largely unclear. We hypothesize that D-serine, a newly identified endogenous co-agonist of N-methyl-D aspartate (NMDA) receptor, may trigger excitotoxicity and neuronal damage in epileptogenesis. By using a mouse pilocarpine model, immunohistochemistry, Fluoro Jade staining and double-labeling, the present study revealed up-regulation of D serine expression in a proportion (41%) of neurons in the cerebral cortex and hippocampus. The D-serine-positive neurons occurred at 4 h, reached peak levels at 12-24 h, and gradually went down at 3-14 days. Moreover, most of D-serine positive neurons were GABAergic (98%), underwent degenerating death (93%), and were accompanied enhancing phosphorylation of NMDA receptor subunit 1. This study has provided new evidence that up-regulation of D-serine production might induce GABAergic neuronal degeneration through excitotoxic mechanism in the pilocarpine model and may be involved in early pathogenesis and recurrent seizure of chronic epilepsy. PMID- 19123039 TI - Primary CNS lymphoma with bilateral symmetric hypothalamic lesions presenting with panhypopituitarism and diabetes insipidus. AB - We present an unusual case of primary central nervous system (CNS) lymphoma presenting with bilateral symmetric hypothalamic lesions causing diabetes insipidus and hypopituitarism. A 50-year-old male presented initially with mental status changes, polyuria and polydipsia. The patient was determined to have diabetes insipidus (DI) and significant anterior pituitary deficiencies resulting in symptomatic pleural and pericardial effusions. Brain MRI with contrast demonstrated bilateral enhancement of his hypothalamus extending to the optic tract. The extensive diagnostic workup that ensued on his initial presentation was non-diagnostic as he had no obvious site of involvement that was easily accessible to biopsy. With close follow-up, the patient had rapid radiographic progression of his disease to his cerebral hemispheres. He therefore underwent brain biopsy and was diagnosed with primary CNS large B cell lymphoma. Chemotherapy has resulted in disease remission with resolution of MRI findings, but the patient has not had resolution of the hypopituitarism or DI. This case highlights the unique diagnostic challenge of patients with isolated hypothalamic lesions. PMID- 19123040 TI - Bioaccumulation of heavy metals in fish tissues of a freshwater lake of Bhopal. AB - Contamination of heavy metals, namely, lead, cadmium, zinc, nickel, copper, chromium and mercury was evaluated in the samples of water and tissues of Labeo rohita and Ctenopharyngodon idella of Upper Lake of Bhopal collected during summer, rainy and winter seasons of 2005-2006. Different organs of the fishes accumulated varying quantities of different heavy metals. In L. rohita, accumulation of heavy metals was in the sequence liver>kidney>gills>muscles, and in C. idella, it was gills>liver>kidney>muscles. Zn was the highest accumulating metal in fish, whilst Hg was the lowest and was well corroborated with those of water. The values of heavy metals were so far well within the maximum permissible standard value of heavy metals for drinking water and for fish culture as prescribed by various national and international agencies. PMID- 19123041 TI - Identification of sources of elevated concentrations of polycyclic aromatic hydrocarbons in an industrial area in Tianjin, China. AB - The concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) were determined by gas chromatography equipped with a mass spectrometry detector in 105 topsoil samples from an industrial area around Bohai Bay, Tianjin in the North of China. Results demonstrated that concentrations of PAHs in 104 soil samples from this area ranged from 68.7 to 5,590 ng g(-1) dry weight with a mean of summation Sigma16PAHs 814 +/- 813 ng g(-1), which suggests that there exists mid to high levels of PAH contamination. The concentration of summation Sigma16PAHs in one soil sample from Tianjin Port was exceptionally high (48,700 ng g(-1)). Ninety three of the 105 soil samples were considered to be contaminated with PAHs (>200 ng g(-1)), and 25 were heavily polluted (>1,000 ng g(-1)). The sites with high PAHs concentration are mainly distributed around chemical industry parks and near highways. Two low molecular weight PAHs, naphthalene and phenanthrene, were the dominant components in the soil samples, which accounted for 22.1% and 10.7% of the summation Sigma16PAHs concentration, respectively. According to the observed molecular indices, house heating in winter, straw stalk combustion in open areas after harvest, and petroleum input were common sources of PAHs in this area, while factory discharge and vehicle exhaust were the major sources around chemical industrial parks and near highways. Biological processes were probably another main source of low molecular weight PAHs. PMID- 19123042 TI - Impact of hemostatic gene single point mutations in patients with non-diabetic coronary artery disease. AB - Single point mutations in the genes coding for hemostatic factors were shown to be major inherited predisposing factors for venous thromboembolism. However, their contribution in the development of non-diabetic coronary artery disease [nDCAD] remains controversial. Angiographically demonstrated nDCAD patients (n = 86) and healthy controls (n = 90) were included in the study. Genotype analysis of hemostatic gene polymorphisms were assessed by using CVD strip assay, based on allele specific oligonucleotide probes. The carrier frequency of factor V (FV) H1299R, prothrombin G20210A, glycoprotein (Gp) IIIa L33P, plasminogen activator inhibitor-I (PAI-1) 4G/5G, 4G/4G, 5G/5G, methylenetetrahydrofolate reductase (MTHFR) A1298C and beta-fibrinogen -455 G > A were similar between patients and controls. In contrast, frequency of FV Leiden was significantly higher among patients (12.5%) than controls (5%, OR: 7.94; 95%CI: 1.9-49.6) and FXIII V34L was significantly lower among patients (23.7%) than controls (40%, OR: 0.24; 95%CI: 0.1-0.89). In addition, the frequency of the MTHFR C677T polymorphism was 32.5% among patients compared with 42.5% in controls, of which the T/T genotype was significantly lower among patients (5%) than controls (17.5%, OR: 0.06; 95%CI: 0.01-0.58). No difference was observed in prevalence of prothrombin G20210A, FV H1299R, Gp IIIa L33P, PAI-1 4G5G, MTHFR A1298C, beta fibrinogen 455 G > A mutations between patients and controls. However, lower frequency of FXIII Val34Leu and MTHFR C677T polymorphisms may decrease, while FV Leiden polymorphism may increase development of nDCAD. PMID- 19123043 TI - The full length cloning of a novel porcine gene CFL2b and its influence on the MyHC expression. AB - Porcine CFL2b gene play an important role in the muscle development and myofibrillar formation in pig. To explore whether CFL2b expression affects muscle fiber trait, the porcine CFL2b full-length cDNA was amplified using homology based cDNA cloning and SMART RACE. Then the full length cDNA of porcine CFL2b was inserted into pEGFP-N1 and transfected into C2C12 cells. The cells stably expressing CFL2b were selected by G418. We examined the expression of MyHC 2x, MyHC 2b and MyHC1/slow in C2C12 cells stably expressing CFL2b. The results showed that the level of MyHC 2x and MyHC 2b mRNA were dramatically increased compared with control cells, while the level of MyHC1/slow mRNA is not changed. To identify the transcription events of CFL2b, the porcine CFL2b mRNA was detected by Northern blotting, two transcripts, long transcript (3,012 bp) and short transcript (1,466 bp) were found in porcine skeletal muscles. The nucleotide sequence of CFL2b shares 88.1 and 74.9% homology with the CFL2b gene in human and mouse. The deduced amino acid sequence of CFL2b (166 amino acids) in pig shares 100, 99.1% identity with the CFL2b in human and mouse, respectively. Taken together, our research revealed that porcine CFL2b may be involved in the regulation muscle fiber trait by affecting the expression of MyHC. PMID- 19123046 TI - Complete nucleotide sequence of chili leaf curl virus and its associated satellites naturally infecting potato in Pakistan. PMID- 19123044 TI - Haplotype and quantitative transcript analyses of Portuguese breast/ovarian cancer families with the BRCA1 R71G founder mutation of Galician origin. AB - We investigated the functional effect of the missense variant c.211A>G (R71G) localized at position -2 of exon 5 donor splice site in the BRCA1 gene and evaluated whether Portuguese and Galician families with this mutation share a common ancestry. Three unrelated Portuguese breast/ovarian cancer families carrying this variant were studied through qualitative and quantitative transcript analyses. We also evaluated the presence of loss of heterozigosity and the histopathologic characteristics of the carcinomas in those families. Informative families (two from Portugal and one from Galicia) were genotyped for polymorphic microsatellite markers flanking BRCA1 to reconstruct haplotypes. Qualitative RNA analysis revealed the presence of two alternative transcripts both in carriers of the BRCA1 R71G variant and in controls. Semi-quantitative fragment analysis and real-time RT-PCR showed a significant increase of the transcript with an out of frame deletion of the last 22nt of exon 5 (BRCA1 Delta22ntex5) and a decrease of the full-length transcript (BRCA1-ex5FL) in patients carrying the R71G mutation as compared to controls, whereas no significant differences were found for the transcript with in frame skipping of exon 5 (BRCA1-Deltaex5). One haplotype was found to segregate in the two informative Portuguese families and in the Galician family. We demonstrate that disruption of alternative transcript ratios is the mechanism causing hereditary breast/ovarian cancer associated with the BRCA1 R71G mutation. Furthermore, our findings indicate a common ancestry of the Portuguese and Galician families sharing this mutation. PMID- 19123047 TI - CARE1, a TY3-gypsy like LTR-retrotransposon in the food legume chickpea (Cicer arietinum L.). AB - We report a Ty3-gypsy like retrotransposon CARE1 (Cicer arietinum retro-element 1) in chickpea (Accession no. DQ239702). This 5,920-bp AT-rich (63%) element carries 723-bp 5'-LTR and 897-bp 3'-LTR flanking to an internal region of 4,300 bp. The LTRs of CARE1 show 93.9% nucleotide identity to each other and have 4-bp (ACTA) terminal inverted repeats. A 17-bp potential tRNA(met) primer binding site downstream to 5'-LTR and a 13-bp polypurine tract upstream to 3'-LTR have been identified. The order of characteristic domains (Gag-proteinase-reverse transcriptase-RNaseH-integrase) in the deduced amino acid sequence and its phylogenetic analysis with other retrotransposons, places CARE1 in the gypsy group of retrotransposons. Homologues of a number of cis-elements including CCAAT, TATA and GT-1 have been detected in the regulatory region or the 5'-LTR of CARE1. Transgenic tobacco plants containing 5'-LTR:GUS construct showed that its 5'-LTR is inactive in a heterologous system under normal as well as tissue culture conditions. Genomic Southern blot experiments using 5'-LTR of the element as a probe showed that CARE1 or its related elements are present in the genomes of various chickpea accessions from various geographic regions. PMID- 19123048 TI - Watershed land use as a determinant of metal concentrations in freshwater systems. AB - Concentrations of Fe, Mn, Cu, dissolved organic matter (DOM), and pH were synthesized from 30 publications to determine the factors regulating concentrations and behavior of metals in freshwater systems. Results from the review suggest that contrasting watershed land use can directly (erosion and runoff) and indirectly (in-lake processes including metal-DOM-pH interactions) affect the metal concentrations in freshwater systems. Among the watershed land uses considered here, concentrations of Fe, Mn, and Cu were observed in the following order: arctic lakes < forested < agricultural < urbanized < mined. A drastic difference in mean metal concentrations has been observed when undisturbed or low impact watersheds (arctic and forested) were changed by agricultural, urban, and mining developments. Relationships between metal concentrations and pH revealed that metals precipitate at high pH (pH > 5). Additionally, at pH < 5, metal concentrations were significantly correlated with DOM due to metal-DOM complexation. High ratios of metal: DOM occur only at low DOM concentrations. Collectively, two general conclusions can be drawn from this review. First, lakes, rivers, and streams with urbanized watersheds are the most susceptible to increased concentrations of metals. Secondly, these results also suggest that regardless of high or low DOM in the water column, pH would affect metal concentrations in freshwater systems. Nonetheless, free metal ions would be higher in freshwater systems with acidic water and low DOM. PMID- 19123049 TI - The involvement of upregulation and translocation of phospho-Rb in early neuronal apoptosis following focal cerebral ischemia in rats. AB - The aim of this study was to investigate the temporal and spatial relationship between phospho-Rb (ser 795) and neuronal apoptotic death in rats subjected to transient focal cerebral ischemia. We found increased phosphorylation of Rb and translocation from neuronal nucleus to cytoplasm in the penumbra zone at 12 h, 1 day, 3 days and 7 days after middle cerebral artery occlusion (MCAO)/reperfusion, compared with sham-operated controls. At 12 h and 1 day, phospho-Rb appeared to be colocalizated with TUNEL staining in neurons, but staining was not colocalizated at 3 days and 7 days. These results demonstrated that cytoplasmic translocation of phospho-Rb from nucleus of neurons occurs in potential apoptotic neurons in the early stages of ischemia/reperfusion, suggesting that the Rb pathway may only be involved in early neuronal apoptosis and may be not an apoptotic signal in the late stages of transient cerebral ischemia. PMID- 19123050 TI - MDR1 (ABCB1) G1199A (Ser400Asn) polymorphism alters transepithelial permeability and sensitivity to anticancer agents. AB - PURPOSE: P-glycoprotein (P-gp), encoded by MDR1 (or ABCB1), is important in anticancer drug delivery and resistance. We evaluated alterations in P-gp mediated transport of anticancer agents due to the MDR1 G1199A polymorphism. METHODS: Using stable recombinant epithelial cells expressing wild-type (MDR1 (wt)) or G1199A (MDR1 (1199A)), anticancer drug sensitivity and transepithelial permeability were evaluated. RESULTS: The recombinant cells MDR1 (wt) and MDR1 ( 1199A ) displayed comparable doxorubicin resistance. However, MDR1 (1199A) cells displayed greater resistance to vinblastine, vincristine, paclitaxel, and VP-16 (11-, 2.9-, 1.9-, and 2.9-fold, respectively). Alterations in transepithelial permeability paralleled these changes. Efflux of doxorubicin was similar between MDR1 wt - and MDR1 (1199A)-expressing cells, while P-gp-mediated transport was greater for vinblastine and vincristine in MDR1 (1199A) cells (2.9- and 2.0-fold, respectively). CONCLUSIONS: The occurrence and magnitude of the MDR1 G1199A effect is drug specific. Overall, the MDR1 G1199A polymorphism may impact anticancer efficacy through modulation of drug distribution and delivery to target tumor cells. PMID- 19123051 TI - Phase II study of paclitaxel combined with capecitabine as second-line treatment for advanced gastric carcinoma after failure of cisplatin-based regimens. AB - PURPOSE: To determine the safety and the efficacy of paclitaxel and capecitabine as second-line combination chemotherapy after failure of platinum regimens in advanced gastric cancer. METHODS: Patients with histologically proven gastric cancer and measurable metastatic disease received capecitabine 825 mg/m(2) twice daily (1,650 mg/m(2) per day) on days 1-14 and paclitaxel 175 mg/m(2) by intravenous infusion on day 1 every 3 weeks until disease progression or unacceptable toxicities. RESULTS: Between June 2003 and October 2005, 26 patients, of median age 59 years (range 41-84 years) were included in the study and were treated by paclitaxel/capecitabine combination. Overall response rate was 34.6% (95%CI = 17.2-55.7%) with one complete response and 42.3% (95%CI = 17.2 55.7%) of patients achieved a stable disease. Median progression-free survival was 4.5 months (95%CI = 4-4.5 months). Median overall survival was 7.5 months (95%CI = 6-10 months). Cumulated overall survival including cisplatin regimens was 15.5 months (95%CI = 11-18 months). Grade 3/4 adverse events included alopecia (30.8%), neutropenia (11.5%), hand foot skin reaction (11.5%), neuropathy (11.5%), arthralgias (7.5%), and anemia (3.8%). CONCLUSIONS: Paclitaxel and capecitabine combination was safe and effective in advanced gastric cancer after failure of cisplatin regimens. The cumulated overall survival of 15.5 months suggests a particular interest of taxanes in second-line treatment after failure of platinum salts. PMID- 19123052 TI - Effect of age on cytokine response in an experimental model of osteomyelitis. AB - To study the effect of age on cytokine response in an experimental model of osteomyelitis. Forty adult male Wistar rats received a stainless steel needle, intramedullarly in the left tibia. Young rats (3 months old) and old rats (22 months old) were allotted in: Group A: Sterile implant. Group B: Sterile implant + slime producing S. aureus. Rats were sacrificed 9 weeks after surgery. Determinations: Cytokines (ELISA) in blood and in tibia extract and the number of bacteria in tibia and implant. The Wilcoxon, Mann-Whitney U tests were used (P < or = 0.01 significant). Infection was detected in every old rat receiving S. aureus, and in 7 of 10 young rats. In blood: prior to surgery, old rats presented higher IL-2 and lower IL-4 levels. Surgery alone did not induce significant changes in old rats; surgery + S. aureus induced significant increases of IL-2 and IL-10 in young rats, and of IL-6 in old rats. Tibia analysis S. aureus group showed increased levels of: IL-10 in young rats, and IL-1beta in old rats. In experimentally induced osteomyelitis, significant differences were observed in cytokine response with regard to age. PMID- 19123053 TI - Gaussian process: an alternative approach for QSAM modeling of peptides. AB - Different statistical modeling methods (SMMs) are used for nonlinear system classification and regression. On the basis of Bayesian probabilistic inference, Gaussian process (GP) is preliminarily used in the field of quantitative structure-activity relationship (QSAR) but has not yet been applied to quantitative sequence-activity model (QSAM) of biosystems. This paper proposes the application of GP as an alternative tool for the QSAM modeling of peptides. To investigate the modeling performance of GP, three classical peptide panels were used: Angiotensin-I converting enzyme inhibitory dipeptides, bradykinin potentiating pentapeptides and cationic antimicrobial pentadecapeptides. On this basis, we made a comprehensive comparison between the GP and some widely used SMMs such as PLS, artificial neural network (ANN) and support vector machine (SVM), and gave the conclusions as follow: (1) for those of structurally complicated peptides, particularly the polypeptides, linear PLS was incapable of capturing all dependences hidden in the peptide systems, (2) even in assistance with the monitoring technique, ANN was inclined to be overtrained in the cases of insufficient number of peptide samples, (3) SVM and GP performed best for the three peptide panels. Moreover, since GP was able to correlate the linear and nonlinear-hybrid relationship, it was slightly superior to SVM at most peptide sets. PMID- 19123054 TI - Implementation of food frequency questionnaire for the assessment of total dietary arsenic intake in Bangladesh: part B, preliminary findings. AB - Dietary intake of water and food has been identified as one of the major pathways for arsenic (As) exposure in the rural population of Bangladesh. Therefore, realistic assessment and measurement of dietary intake patterns are important for the development of an accurate estimate of As exposure and human health risk assessment. One important consideration is to identify an appropriate tool for measuring dietary intake. In this study an interviewer-administered Food Frequency Questionnaire (FFQ) was implemented to determine age and gender specific dietary intake. The developed FFQ was unique because it developed a synergy between field dietary assessment and As concentration measurements in various environmental media. The resulting integrated database provided an accurate framework for the process of As exposure and human health risk assessment. The preliminary results reported here from the FFQ demonstrated that this technique could be used in rural areas as a tool to assess As exposure and the associated human health risk. PMID- 19123056 TI - Predicting lung cancer death in Africa and Asia: differences with WHO estimates. AB - OBJECTIVE: Reliable cancer burden estimates are rarely available from most developing countries where cancer registration is lacking. This study provided estimates on the current and future number of lung cancer deaths in Indonesia, Vietnam and Ethiopia, and Sub-Saharan Africa at large. METHODS: The number of lung cancer deaths was estimated from detailed smoking prevalence data (obtained from surveys among 8,726 rural individuals aged 25-74 years in Indonesia, Vietnam, and Ethiopia in 2005-2006) and on lung cancer rate estimates among non smokers. RESULTS: Our estimate for lung cancer deaths in Sub-Saharan Africa is 44,076 in 2005, which is 2.6 times the most recent WHO estimate in 2003 (17,000 deaths). A similar ratio is found for the country-specific estimate in Ethiopia. Our estimates are only slightly higher than the WHO's in Indonesia, and Vietnam. The attributable risk of smoking for lung cancer death among men was 39% in Ethiopia, 80% in Indonesia and 85% in Vietnam. We expect the annual number of lung cancer deaths to double by 2025, even if the smoking prevalence is assumed not to increase further. CONCLUSIONS: WHO estimates on lung cancer deaths in Asia appear to be slightly lower than our study results; however, in Africa, the burden appears to be largely underestimated. PMID- 19123055 TI - Physical activity and function in older, long-term colorectal cancer survivors. AB - OBJECTIVE: Increasing age and cancer history are related to impaired physical function. Since physical activity has been shown to ameliorate age-related functional declines, we evaluated the association between physical activity and function in older, long-term colorectal cancer survivors. METHODS: In 2006-2007, mailed surveys were sent to colorectal cancer survivors, aged > or = 65 years when diagnosed during 1995-2000, and identified through a state cancer registry. Information on physical activity, physical function, and relevant covariates was obtained and matched to registry data. Analysis of covariance and linear regression were used to compare means and trends in physical function across levels of activity in the final analytic sample of 843 cases. RESULTS: A direct, dose-dependent association between physical activity and function was observed (p(trend) < .001), with higher SF-36 physical function subscores in those reporting high versus low activity levels (65.0 +/- 1.7 vs. 42.7 +/- 1.7 (mean +/ standard error)). Walking, gardening, housework, and exercise activities were all independently related to better physical function. Moderate-vigorous intensity activity (p(trend) < .001) was associated with function, but light activity (p(trend) = 0.39) was not. CONCLUSION: Results from this cross-sectional study indicate significant associations between physical activity and physical function in older, long-term colorectal cancer survivors. PMID- 19123058 TI - Review and implementation of cure models based on first hitting times for Wiener processes. AB - The development of models and methods for cure rate estimation has recently burgeoned into an important subfield of survival analysis. Much of the literature focuses on the standard mixture model. Recently, process-based models have been suggested. We focus on several models based on first passage times for Wiener processes. Whitmore and others have studied these models in a variety of contexts. Lee and Whitmore (Stat Sci 21(4):501-513, 2006) give a comprehensive review of a variety of first hitting time models and briefly discuss their potential as cure rate models. In this paper, we study the Wiener process with negative drift as a possible cure rate model but the resulting defective inverse Gaussian model is found to provide a poor fit in some cases. Several possible modifications are then suggested, which improve the defective inverse Gaussian. These modifications include: the inverse Gaussian cure rate mixture model; a mixture of two inverse Gaussian models; incorporation of heterogeneity in the drift parameter; and the addition of a second absorbing barrier to the Wiener process, representing an immunity threshold. This class of process-based models is a useful alternative to the standard model and provides an improved fit compared to the standard model when applied to many of the datasets that we have studied. Implementation of this class of models is facilitated using expectation maximization (EM) algorithms and variants thereof, including the gradient EM algorithm. Parameter estimates for each of these EM algorithms are given and the proposed models are applied to both real and simulated data, where they perform well. PMID- 19123057 TI - Upregulation of BACE1 and beta-amyloid protein mediated by chronic cerebral hypoperfusion contributes to cognitive impairment and pathogenesis of Alzheimer's disease. AB - Chronic cerebral hypoperfusion (CCH) increases the risk of Alzheimer disease (AD) through several biologically plausible pathways, but the relationship between CCH and the development of AD remains uncertain. To investigate expression of APP, BACE1 and A beta in the hippocampus of BCCAO rats and study pathophysiological mechanism of AD from CCH. CCH rat model was established by chronic bilateral common carotid artery occlusion (BCCAO). Behavior was evaluated after BCCAO with Morris water maze and open-field task. Expression of A beta was measured by enzyme linked immunosorbent assay (ELISA). beta-Amyloid precursor protein cleavage enzyme 1 (BACE1) and beta-amyloid precursor protein (APP) were tested by ELISA, Western blotting and RT-PCR. Cognitive impairment occurred with CCH by Morris water maze test and open-field task. The BACE1 and A beta level in BCCAO rats was more increased than sham-operation control rats (P < 0.01) but APP had no difference(P > 0.05). The expression of BACE1 and A beta has no inter-group difference in BCCAO rats (P > 0.05). The level of BACE1 and A beta had positive correlation with cognitive impairment (P < 0.01) while no correlation was observed between APP and cognitive impairment. Chronic cerebral ischemia contributes to cognitive impairment and vascular pathogenesis of Alzheimer's disease that chronic cerebral hypoperfusion increases BACE1 and A beta level in brain. PMID- 19123059 TI - Treatment of type-A3 amyelic thoracolumbar fractures (burst fractures) with kyphoplasty: initial experience. AB - The authors propose a minimally invasive procedure for treating type-A3 amyelic thoracolumbar fractures according to Magerl classification (compression burst fractures). The procedure, percutaneous kyphoplasty, allows the fracture to be reduced and vertebral height to be restored by injecting bone cement into a cavity created in the vertebral body by an inflatable balloon introduced with the percutaneous approach. Four patients were successfully treated, with clinical and functional benefit in all cases. There were no complications. PMID- 19123060 TI - Conivaptan bolus dosing for the correction of hyponatremia in the neurointensive care unit. AB - INTRODUCTION: Hyponatremia frequently complicates acute brain injury and may precipitate neurological worsening by promoting cerebral edema. An increase in brain water may be better managed through water excretion than with fluid restriction or hypertonic fluids. Vasopressin-receptor antagonists such as conivaptan, which promote free water excretion, may be ideal agents to treat this common and potentially serious disorder. METHODS: The efficacy of intermittent bolus doses of conivaptan to correct hyponatremia was examined in a consecutive series of patients treated in our neurointensive care unit. Patients were excluded if baseline sodium was over 135 mEq/l or if another conivaptan dose was given within 12 h. We assessed the proportion responding with a 4 or 6 mEq/l rise in sodium by 12 h, the change in sodium from baseline, and, in those not receiving another dose for at least 72 h, the long-term ability of a single dose to maintain sodium at least 4 mEq/l above baseline. We also recorded the effects of conivaptan on urine output and specific gravity, and noted any adverse events. RESULTS: A total of 25 doses given to 19 patients were included (out of 44 total doses administered in the study period). Sodium rose by 5.8 +/- 3.2 mEq/l within 12 h, with 71% rising by at least 4 mEq/l and 52% manifesting at least a 6 mEq/l increase. In those receiving only a single dose, 69% maintained at least a 4 mEq/l rise up to 72 h. Conivaptan also consistently led to increased urine output and a significant drop in urine specific gravity (i.e., aquaresis). No cases of phlebitis were observed despite administration of conivaptan through peripheral IVs. CONCLUSION: Intermittent dosing of conivaptan was effective in increasing free water excretion and correcting hyponatremia in neurologically ill patients. This supports its further evaluation for managing hyponatremia in this population. PMID- 19123061 TI - The positive force-frequency relationship is maintained in absence of sarcoplasmic reticulum function in rabbit, but not in rat myocardium. AB - Myocardial calcium handling differs between species, mainly in the relative contribution between the sources for activator calcium. To investigate the role of the myofilaments and intracellular calcium decline in governing the relaxation phase of cardiac muscle, and to elucidate additional determinants of relaxation other than the sarcoplasmic reticulum (SR) at various frequencies within the in vivo range, the present study was performed by altering the calcium handling in rat and rabbit. Trabeculae, iontophoretically loaded with bis-fura-2 to monitor cytoplasmic calcium levels, were subjected to ryanodine and cyclopiazonic acid to inhibit SR function. Simultaneous force and [Ca(2+)](i) measurements were obtained at 1-4 Hz in rabbit and at 4-8 Hz in rat before and after SR inhibition. Inhibition of the SR resulted in increased diastolic and peak calcium levels as well as decreased developed force in both species. Calcium transient amplitude decreased in rat, but increased in rabbit after SR inhibition. Time to peak tension, time from peak tension to 50% relaxation, time to peak calcium, and time from peak calcium to 50% calcium decline were all prolonged. Results suggest that L-type calcium channel current is responsible for increases in calcium with increasing frequency, and that the SR amplifies this effect in response to increased L-type current. The response of the myofilaments to alterations in calcium handling plays a critical role in the final determination of force, and may differ between species. These results imply the balance between force relaxation and calcium decline is significantly different in larger mammals, necessitating a critical re-evaluation of how myocardial relaxation is governed, specifically regarding frequency-dependent activation. PMID- 19123062 TI - Gas exchange and photosynthetic performance of the tropical tree Acacia nigrescens when grown in different CO(2) concentrations. AB - The photosynthetic responses of the tropical tree species Acacia nigrescens Oliv. grown at different atmospheric CO(2) concentrations-from sub-ambient to super ambient-have been studied. Light-saturated rates of net photosynthesis (A (sat)) in A. nigrescens, measured after 120 days exposure, increased significantly from sub-ambient (196 microL L(-1)) to current ambient (386 microL L(-1)) CO(2) growth conditions but did not increase any further as [CO(2)] became super-ambient (597 microL L(-1)). Examination of photosynthetic CO(2) response curves, leaf nitrogen content, and leaf thickness showed that this acclimation was most likely caused by reduction in Rubisco activity and a shift towards ribulose-1,5-bisphosphate regeneration-limited photosynthesis, but not a consequence of changes in mesophyll conductance. Also, measurements of the maximum efficiency of PSII and the carotenoid to chlorophyll ratio of leaves indicated that it was unlikely that the pattern of A (sat) seen was a consequence of growth [CO(2)] induced stress. Many of the photosynthetic responses examined were not linear with respect to the concentration of CO(2) but could be explained by current models of photosynthesis. PMID- 19123063 TI - Different modulation of the outputs of yeast MAPK-mediated pathways by distinct stimuli and isoforms of the dual-specificity phosphatase Msg5. AB - The activity of protein phosphatases on mitogen-activated protein kinases (MAPKS) is essential in the modulation of the final outcome of MAPK-signalling pathways. The yeast dual-specificity phosphatase (DSP) Msg5, expressed as two isoforms of different length, dephosphorylates the MAPKs of mating and cell integrity pathways, Fus3 and Slt2, respectively, but its action on the MAPK Kss1 is unclear. Here we analyse the global impact of Msg5 on the yeast transcriptome. Both Fus3- and Slt2- but not Kss1-mediated gene expression is induced in cells lacking Msg5. However, although these cells show high Slt2 phosphorylation, the Rlm1-dependent Slt2-regulated transcriptional response is weak. Therefore, mechanisms concomitant with Slt2 phosphorylation are required for a strong Rlm1 activation. The limited Slt2 activity on Rlm1 is not a specific effect on this substrate but a consequence of its low kinase activity in msg5Delta cells. Lack of Msg5 does not increase Kss1 phosphorylation although both proteins physically interact. Both Msg5 isoforms interact similarly with Slt2, whereas the long form binds Fus3 with higher affinity and consequently down-regulates it more efficiently than the short one. We propose that specific binding of DSP isoforms to distinct MAPKs provides a novel mechanism for fine tuning different pathways by the same phosphatase. PMID- 19123064 TI - Thermoelectric effect on charged colloids in the Huckel limit. AB - We study the thermophoretic coefficient D(T) of a charged colloid. The non uniform electrolyte is characterized in terms of densities and diffusion currents of mobile ions. The hydrodynamic treatment in the vicinity of a solute particle relies on the Huckel approximation, which is valid for particles smaller than the Debye length, a << [Formula: see text] . To leading order in the parameter a/[Formula: see text] , we find that the coefficient D(T) consists of two contributions, a dielectrophoretic term proportional to the permittivity derivative dvarepsilon/dT , and a Seebeck term, i.e., the macroscopic electric field induced by the thermal gradient in the electrolyte solution. Depending on the particle valency, these terms may take opposite signs, and their temperature dependence may result in a change of sign of thermophoresis, as observed in several recent experiments. PMID- 19123065 TI - Influence of narcotics on luminance and frequency modulated visual evoked potentials in rats. AB - Quantification of visual function is essential for the impact of disease models and their treatment. Recently, we introduced a chronic implant model to record visual evoked potentials (VEP) in awake Brown-Norway rats. Here, we investigated the hemispheric distribution of VEP after monocular stimulation, the chronic electrode implantation and the influence of commonly used anesthetics. Potentials were recorded by electrodes, implanted epidurally over the superior colliculus. The entire visual field of the rat was stimulated. Flicker stimuli were modulated in luminance with a modulation depth from 5 to 80% at 7.5 Hz and flashes with a modulation depth of >95% in a frequency range of 2.9-38 Hz. Recordings were constant over 9 days. When one eye was blinded, the potentials recorded from the contralateral side were not affected, while the potentials of the ipsilateral side were markedly reduced. Further, potentials of awake animals were compared with those receiving general anesthetics. For one group of rats (n = 8), we administered isoflurane by inhalation in five concentrations. Four different groups (n = 7-11) were given choralhydrate (200 and 400 mg/kg) and the combination of ketamine/xyaline (65/7 or 130/14 mg/kg, respectively) intraperitoneally. Isoflurane depressed the VEP in a concentration-dependent manner. Treatment with chloralhydrate and ketamine/xyaline increased the VEP at low concentrations and depressed it in high concentrations. The new VEP paradigm assesses distinct qualities of contrast vision in rats. All tested narcotics alter VEP amplitudes and can be avoided. PMID- 19123066 TI - Ganoderma lucidum (Fr.) P. Karst enhances activities of heart mitochondrial enzymes and respiratory chain complexes in the aged rat. AB - Aging is associated with increased oxidative damage at multiple cellular levels, decline in cellular energy production and enhanced free radical status. The effect of the medicinal mushroom, Ganoderma lucidum on the activities of tricarboxylic acid (Krebs) cycle enzymes and mitochondrial complexes I-IV of the electron transport chain in aged rats were investigated. The activity of Krebs cycle enzymes, isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, and malate dehydrogenase as well as mitochondrial complexes I, II, III, and IV were determined in heart of aged male Wistar rats orally administrated with 70% ethanolic extract (50 and 250 mg/kg) of G. lucidum. DL-alpha-lipoic acid (100 mg/kg) was taken as the positive control. Administration of the G. lucidum, once daily for 15 days, was significantly (P < 0.05) effective to enhance the Krebs cycle dehydrogenases, and mitochondrial electron transport chain complex IV activities in aged rats. The profound activity of the extract can be correlated to the significant antioxidant property of G. lucidum. The results of the study revealed that G. lucidum is effective to ameliorate the age associated decline of cellular energy status. PMID- 19123067 TI - Abstracts of the ESCP 37th European Symposium on Clinical Pharmacy, Pharmaceutical Care Models, and Therapeutic Innovations. Dubrovnik, Croatia. October 21-24, 2008. PMID- 19123068 TI - Inducible expression of human angiostatin by AOXI promoter in P. pastoris using high-density cell culture. AB - A high-density cell culture method was successfully established in P. pastoris with the alcohol oxidase I (AOXI) promoter in order to produce large quantities of recombinant human angiostatin (AS) which has been reported to have antiangiogenic activity. A preliminary study on fermentation conditions in shaking flasks indicated that adequacy of biomass is beneficial to obtain more products. The fermentation was carried out in a 10 l bioreactor with 5 l modified growth medium recommended by Invitrogen at 30 degrees C. The cells were first grown in glycerol-PTM4 trace salts for 24 h. When the cell density reached A(600) = 125, methanol-PTM4 trace salts was added to induce the expression of AS. During the fermentation, dissolved oxygen level was maintained at 20-30%, pH was controlled at 5 by the addition of 7 M NH(4)OH and the biomass was maintained at about A(600) = 200. After 60 h of induction, the secreted AS was 153 mg/l. The recombinant AS inhibited the angiogenesis on CAM and suppressed the growth of B16 melanoma in C57BL/6J mice (P ?0.01). PMID- 19123069 TI - Influence of nitrogen on the expression of TaDof1 transcription factor in wheat and its relationship with photo synthetic and ammonium assimilating efficiency. AB - Nitrogen is a crucial macronutrient needed in the greatest amount of all mineral elements required by plants. Development of crop varieties with high nitrogen use efficiency (NUE) is imperative for sustainable agriculture. Understanding how plant genes respond to different nitrogen conditions is essential for formulating approaches, for manipulating genes, for improving NUE. In the present study we analyzed the activity of three different enzymes involved in nitrogen assimilation viz., GS, GOGAT and GDH along with physiological parameters like chlorophyll variable yield (Fv/Fmax), photosynthesis rate and total chlorophyll content at four different growth stages of wheat plant development under different nitrogen treatments. For this study two different wheat varieties UP 2644 and Raj-4097 having high and low protein content, respectively in the grains were chosen. Gene expression profile of a Dof transcription factor (TaDof1 of wheat) was also included in the study to assess its role in nitrogen metabolism. Densitometry analysis at S(2) and S(3) stage of wheat spikes of both the wheat varieties grown at different nitrogen treatments showed that TaDof1 expression was up-regulated in low nitrogen treatment. In S(3) stage, in high protein content wheat variety UP-2644, TaDof1 expression was elevated in low and normal nitrogen treatment as compared to high nitrogen treatment. The gene expression profile of Dof 1 was found to coincide with the enzyme activities of GS, GOGAT at the S(3) stage. The activities of these enzymes were prolonged in the high protein content variety. Since, Dof transcription factor(s) have been previously reported to control the expression of genes involved nitrogen assimilation i.e., GS and GOGAT and may be the elevated expression of Dof 1 at the grain filling stage over expresses the GS and GOGAT genes thereby prolonging their activities. PMID- 19123070 TI - Factors associated with short-term recovery of health status among emergency department patients with acute exacerbations of chronic obstructive pulmonary disease. AB - OBJECTIVE: To determine the factors associated with short-term recovery of health status following chronic obstructive pulmonary disease (COPD) exacerbation. METHODS: In a prospective multicenter cohort study, consecutive emergency department (ED) patients with COPD exacerbation were enrolled. Patients were interviewed and instructed to complete the validated Short Form Chronic Respiratory disease Questionnaire. Follow-up data were collected 2 weeks later, and included the global transition question. The primary outcome was recovery of health status, which was assigned to patients who stated that their COPD was "a little better" or "much better" than at the time of their ED presentation. RESULTS: Of the 330 patients, 270 [82%; 95% confidence interval (CI), 77-86%] reported recovery of health status following acute exacerbations. Multivariable analysis showed that recovery of health status was associated with having a primary care provider (PCP) [odds ratio (OR), 3.1; 95% CI, 1.5-6.4] and a co diagnosis of asthma (OR, 2.2; 95% CI, 1.2-3.8). By contrast, frequent exacerbations (two or more exacerbations in the past year) (OR, 0.4; 95% CI, 0.2 0.7) was inversely associated with recovery. CONCLUSIONS: Reducing exacerbation frequency and provision of adequate PCP follow-up after ED visit may help improve the recovery of health status following acute exacerbations of COPD. PMID- 19123071 TI - Breast cancer immunohistochemistry can be useful in triage of some HNPCC families. AB - Immunohistochemistry of tumour samples is increasingly used in the triage of families where hereditary non-polyposis colorectal cancer (HNPCC) due to mismatch repair defects is suspected. Usually, this is undertaken in tumours that are a recognised part of the spectrum of HNPCC-related cancers e.g. colon or endometrial cancers. Although breast cancers are not classed as part of this spectrum, this study examined the extent to which some breast tumours do arise by the mismatch repair pathway in these families. This may have clinical utility in families where an individual with a 'classic HNPPC-related' tumour is not available for evaluation. Immunohistochemistry of a breast tumour may identify an individual in whom germline mutation testing is worthwhile. PMID- 19123072 TI - Does admission from different sources have any influence on intensive care unit outcome in COPD patients? AB - Influence of admission source and admission source related factors on intensive care unit (ICU) outcome have not known much in patients with chronic obstructive pulmonary disease (COPD). The aim of the study was to investigate if admission source and related factors have any impact on ICU outcome in patients with COPD. A retrospective observational cohort study. Demographics of the patients, severity of admission disease and airflow limitation, comorbidity, source of admission [pulmonary ward (PW), emergency department (ED)], noninvasive mechanical ventilation (NIMV) therapies in the pre-and post-intubation period, and blood gases were recorded. T-test, chi-square test and logistic regression analysis were used for statistical analysis. One hundred and four patients were included in the study. Fifty two of them were admitted from PW and 52 from ED. There were no significant difference between age, gender, comorbidity, severity of airflow limitation and admission Acute Physiology Assessment and Chronic Health Evaluation (APACHE)-II scores among the patients admitted from PW and ED. While the patients admitted from ED were more hypoxemic, admitted with community acquired pneumonia more frequently, the patients admitted from PW, received NIMV trial in pre-intubation and post-extubation period more frequently (p<0.001) and had higher HCO-(3) levels. There was no significant difference in the ICU survival across the groups. Among these potential risk factors higher APACHE-II scores and duration of mechanical ventilation were independent risk factors for the mortality. These results suggest that while some of the admission characteristics were significantly different, admission from ED or PW did not have negative influence on ICU course and outcome in patients with COPD. PMID- 19123073 TI - [The importance of the at risk COPD patients (Stage 0) and clinical differences]. AB - Chronic obstructive pulmonary disease (COPD) is an important world health problem and estimated that the prevalance will increase. In early period this disease in symptomatic with cough, sputum and dyspnea. FEV(1)/FVC was greater than 70% in early period and it was accepted as stage 0 (at risk) COPD before the last The Global Initiative for Chronic Obstructive Lung Disease (GOLD) guideline. In last GOLD guideline this stage is excluded. In this study we aimed to find out the clinical differences between this cases and healty population. Prospectively spirometric measures, exercise capacity, dyspnea index, saturation of O2 (SpO2) and life quality are compared between two groups. We evaluated 58 at risk cases and 63 healty adults. There was no difference between two groups about age, sex and smoke cessations. In at risk group 77.6% cough, 53.4% sputum and 62.1% dyspnea were the symptoms. In currently smokers cough and sputum were dominant symptoms however dyspnea was dominant symptom in exsmokers. Absolute FEV1, FEF25 75 values, predicted FEV1/FVC, FEV1, FVC, PEF, FEF25-75% values are lower in at risk group (p<0.05). Mean six minutes walking distance were lower in at risk group but not statistically significant (p>0.05). Dyspnea index before walking were not different between two groups (p>0.05) however dyspnea index after walking and change were higher in at risk group (p>0.05). Resting SpO2, SpO2 after exercise and desaturation with exercise (DeltaSpO2) were lower in at risk group but not statistically significant. Physical and mental components of Short Form 36 (SF36) scores were similar in two groups (p>0.05). COPD at risk group is separated from healty population with chronic respiratory symptoms and inhalation of smoke or other irritans. In these cases spirometric measures are significantly worse although functional parametres are similar with healty population, so it is thought about being a subclinical disease. These cases must be educated about COPD, encouraged about giving up smoking and prevented occurring COPD. PMID- 19123074 TI - The prognostic value of serum epidermal growth factor receptor level in patients with non-small cell lung cancer. AB - Epidermal growth factor receptor (EGFR) has been implicated as a factor indicating tumour progression or as a prognostic factor in non-small cell lung cancer (NSCLC), in which its overexpression is often detected. The usefulness of identifying EGFR in serum from patients with NSCLC is controversial. This study was designed to identify serum EGFR levels in patients with NSCLC and to evaluate the relationship between serum EGFR levels and clinical stage, histological subtype and survival time. Serum EGFR levels were measured using quantitative enzyme-linked immunosorbent assay. The study included 43 patients with NSCLC and 16 healthy controls. The histological classification was 29 squamous carcinomas and 14 adenocarcinomas. Serum samples were collected before treatment.There was no difference between serum EGFR levels in patients with NSCLC (17.53+/-8.09 fmol/mL) in comparison with those healthy controls (16.88+/-7.08 fmol/mL; p=0.912). There was also no difference in serum EGFR levels according to clinical stage or histological subtype. There was no relationship between serum EGFR levels and survival time in patients with NSCLC. The study's results suggest that, the utility of serum EGFR levels in patients with NSCLC as a tumour marker or as a prognostic factor is limited. However, further prospective studies on a large number of patients will be necessary to confirm this study's results. PMID- 19123075 TI - [Evaluation of tuberculosis cases detected in our region between 1999 and 2003]. AB - This study investigates the extent of tuberculosis, its clinical features, the specification of the patients with tuberculosis, and evaluates diagnosis, treatment and their effectiveness in our region. The records of 645 patients admitted to the Tuberculosis Control Dispensary in our between January 1999 and December 2003 were evaluated retrospectively. The incidence of the disease was 0.012% and it was more common among young men and diagnosed mostly by individual examination. Twenty-six percent of the patients had a history of physical contact with a tuberculosis patient. The pulmonary and extra pulmonary involvement of the disease was 51.8% and 48.2%, respectively. Bacteriologic examination of the patients was as follows; 42.2% did not have a bacteriologic examination, 21.7% was smear and culture positive, 26.8% was smear and culture negative, and 8.7% was smear negative and culture positive. Diagnosis of tuberculosis by bacteriologic examination increased from 39.7% to 73.6% within years. Cure rates of the disease were between 12.5% and 34.4%. Completion rates of tuberculosis therapy were between 53.5% and 72.6% and treatment success rates were between 82% and 95% whereas mortality rates varied between 1.7% and 2.8%. In conclusion, we observed that new cases and smear positive cases of tuberculosis were more common among the young and among the people with low socioeconomic status, bacteriologic examination increased over time and the rate of successful treatment was high but the cure rate was low. PMID- 19123076 TI - [Symptom prevalence of obstructive sleep apnea-hypopnea syndrome in health-care providers in central Sivas]. AB - Obstructive sleep apnea-hypopnea syndrome (OSAHS) is the most common sleep disorder that can lead to serious complications. Polysomnography (PSG) is the gold standard for the diagnosis of OSAHS. Unfortunately, PSG studies are expensive, time-consuming, requiring special team and equipment. Therefore, it is possible to determine the cases likely to have OSAHS requiring at least PSG by type A studies. There isn't enough data about the prevalence of OSAHS in Turkey. The aim of this study was to estimate the symptom prevalence of OSAHS in health care providers in central Sivas. The questionnaire asking demographic features, additional diseases, habits, the symptoms related with OSAHS was performed in total of 1202 health care providers. We also performed Epworth sleepiness scale (ESS) to determine excessive daytime sleepiness. Snoring was detected in 267 participants. Snoring rates were 38% and 10.9% in men and in women, respectively. The rate of witnessed sleep apnea in all cases was 4.4% (n=53). Witnessed sleep apnea was reported by 42 of men (8.4%) and 11 of women (1.6%). A total of 338 participants had excessive daytime sleepiness. There was a significant relation between three major symptoms of OSAHS and excessive sleepiness while driving. All three major symptoms were detected in 22 cases (1.8%), of which 21 were males. This study suggested that OSAHS symptom prevalence is quite high helth-care provides in our region and, therefore expanded usage of PSG studies is required. PMID- 19123077 TI - Oedema risk of endotracheal intubation in rabbits with cessation of steroid therapy. AB - Rebound oedema of tissues is a well defined complication of cessation of steroid therapy. Tapering of systemic corticosteroid regimens in short course steroid therapy is considered unnecessary in most circumstances in acute exacerbation of chronic obstructive pulmonary diseases, presence of laryngeal rebound edema is obscure in this situation. We studied whether or not laryngeal oedema increases after intubation when intubation is established after cessation of steroid therapy. Thirty-six rabbits were randomly divided into six groups. We administered 1 mg/kg methyl prednisolone intraperitoneally to four steroid groups for ten days. Another group received serum physiologic for ten days and last group was sham control that was intubated only. Rabbits that received steroid therapy were intubated and separated into groups one day, one week, two weeks, and a month after the cessation of steroid therapy. Airway area and percentage of cross sectional area of larynx lumen to their own larynx tissues surrounded by thyroid cartilage and oesophagus were studied by stereological methods. Larynx lumen area of one week steroid group was significantly narrower and percentage of cross sectional area of larynx lumen to their own larynx tissues surrounded by thyroid cartilage and oesophagus was significantly larger than sham control. Rebound oedema forms in larynx with abrupt cessation of steroid therapy in rabbits. Clinical safe time for intubation after abrupt cessation of steroid therapy is also defined with our study. These results suggest that one week after the cessation of steroid therapy may be a hazardous time for tracheal intubation. PMID- 19123078 TI - [The evaluation of the pneumoconiosis cases, whose liability times being expired, applied to the Social Security High Health Committee between the years 1998 2001]. AB - This study was planned with the aim to evaluate the pneumoconiosis cases which applied to Social Security High Health Committee between January 1st 1998 and December 31st 2001 for their liability times to be extended. It was a retrospective and descriptive study. Of the 1108 files, only 1068 of them are taken into evaluation with 96.4% of the participation due to the lacking data of some files. The data entry and analysis was done by using SPSS. All of the applicants were male. Most of them applied in 1998 (41.1%, 439 cases) and in 2000 (36.6%, 391 cases). Their average working years were 23.51+/ 0.25. Their average year's of age being diagnosed as pneumoconiosis was 61.81+/-0.8. Of the all cases 764 (72.1%) were diagnosed within the first 10 years after the legal liability time. At this point to reach more conclusive results further epidemiologic studies are required including the factors (the concentration of the dust in the work place besides the physical properties of the dust, the exposure time to the dust and individual properties) affecting the formation of pneumoconiosis. However, even with these findings it is been concluded that the legal time being 10 years in use now for determination of the liability time for the pneumoconiosis cases should be lengthened. It is been also concluded that with the cooperation of other organizations the protocols consisting the issues about the diagnosis criteria of the pneumoconiosis cases, the treatment plan, the determination of disability ratio, liability time, the examinations before getting the job and while working periodically, assessment of the work place risks should be made. Considering the average age being 61.81 for the workers who retired after working dusty work places, carrying the pneumoconiosis risk, they should be checked in regular intervals. PMID- 19123079 TI - The diagnosis of sarcoidosis pleurisy by medical thoracoscopy: report of three cases. AB - Pleural effusion in sarcoidosis is infrequent. The data on thoracoscopic observations of sarcoidosis pleurisy are limited. The present study describes three cases of sarcoid-related pleurisy diagnosed by medical thoracoscopy and discusses the thoracoscopic features of sarcoid pleuritis. The appearance of pleural nodules was completely different in the three cases, and the distribution of nodules of sarcoidosis was heterogenously located in the pleural surfaces. Pleural disease in sarcoidosis could be proved easily by sampling visible nodules, and pleural fluid could be aspirated without complications during thoracoscopy. Due to symptomatic pleurisy of sarcoidosis, therapy was given as systemic corticosteroid. In conclusion, thoracoscopy may be an appropriate alternative technique to obtain an accurate diagnosis in sarcoid pleurisy. PMID- 19123080 TI - [Churg-Strauss syndrome associated with montelukast]. AB - A 47 year old woman who had a history of asthma for 15 years referred to our hospital because of infiltrates on her chest radiograph that not responded to antibiotic treatment. We found that she had eosinophilia in peripheral blood (38%) and bronchoalveolar lavage (54%), nasal polyposis, and transient pulmonary infiltrates, and in the base of these findings we diagnosed as Churg-Strauss Syndrome (CSS). She has been using montelukast for 2 years. By examining her previous medical records, we observed that while eosinophil rates in peripheral blood were normal before montelukast usage, after this therapy eosinophil rates were greater 10 percent. Therefore, we thought that CSS was to be associated with montelukast usage. After just montelukast therapy was discontinued, clinical and radiographic parameters and the eosinophil counts (20%) improved. We present this case of CSS associated with montelukast in whom spontaneous remission was observed without using corticosteroids and cytotoxic agents. PMID- 19123081 TI - Pleural effusion as the first sign of multiple myeloma. AB - Multiple myeloma (MM) is a rare disorder that affects all tissues, except reticuloendothelial tissues, and seldom causes a myelomatous pleural effusion. A 56-year-old male patient attended our clinic complaining of shortness of breath and fatigue for the past month. A posteroanterior chest radiograph revealed a left pleural effusion, which was subsequently assessed as being exudative in nature. Protein electrophoresis demonstrated hypergammaglobulinemia. Serum and pleural fluid immunofixation electrophoresis showed an increase in the IgG and kappa fractions. The Bence-Jones protein level in the 24-hours urine was high. Pleural biopsy showed plasmocyte infiltration. Bone marrow biopsy revealed atypical plasma cell infiltration, some with immature morphology, carrying IgG/Kappa clonality in a diffuse pattern. The patient was diagnosed with IgG/k type MM and underwent chemotherapy with vincristine, doxorubicin, and prednisolone. Complete regression of the pleural effusion was achieved after one round of chemotherapy, and the patient has been followed for 18 months. PMID- 19123082 TI - [A rare agent of pneumonia: Brucella melitensis]. AB - Brucellosis remains a world-wide public health problem especially in developing brucellosis remains a world-wide public health problem especially in developing countries. Although involvement of the respiratory system in brucellosis is an acknowledged but rare event, its clinical manifestations and focal complications are often troublesome in making a diagnosis. Herein, we report a pneumonia case which proved due to Brucella melitensis in a 57-year-old man who presented with chronic cough, sputum, hemoptysis and fever. PMID- 19123083 TI - [Tuberculous pleurisy after tumour necrosis factor-alpha antagonist usage: case report]. AB - A thirty-six year old male patient presented with dyspnea, right-sided chest pain, night sweats and intermittent fever. He has a history of ankylosing spondylitis treated with tumour necrosis factor-alpha (TNF-alpha) antagonist (infliximab). Computed tomography of the chest showed mediastinal lymphadenopathy, right-sided pleural effusion, and atelectasis. The pleural fluid was exudative with lymphocyte dominance. Closed pleural biopsy was nondiagnostic. The adenosine deaminase level of the pleural fluid was 110 U/L. In light of these findings, the patient was diagnosed as tuberculous pleurisy and antituberculous treatment was given. After one month, pleural fluid was markedly reduced. PMID- 19123084 TI - Late presentation of cystic adenomatoid malformation type 4 of lung in an asymptomatic child. AB - Congenital cystic adenomatoid malformation of lung is an uncommon embryonic developmental anomaly usually diagnosed perinatally. Rarely the presentation is delayed until childhood and adulthood. Herein we present a 10-years-old girl with a late presentation of congenital cystic adenomatoid malformation type 4 of lung who was diagnosed coincidentally. PMID- 19123085 TI - Acute anuric renal failure with streptokinase therapy in a patient with acute venous thromboembolic disease and the review of renal side effects of streptokinase. AB - The benefits of thrombolytic therapy in acute myocardial infarction are now well established. However many uncertainties, such as adverse effects, are still remain in venous thromboembolic disease. We describe a unique patient who treated with streptokinase for the methylen tetrahydrofolate reductase mutation associated acute deep vein thrombosis and massive pulmonary embolism. After therapy patient developed acute anuric renal failure without an evidence of bleeding or immunologic reaction and we would like to review the renal side effects of streptokinase in patients with venous thromboembolic disease. PMID- 19123086 TI - Anticlockwise migrating infiltration in both lungs. PMID- 19123087 TI - [Chronic obstructive pulmonary disease and cardiovascular system]. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality worldwide killing nearly 3 million people annually. Even the most optimistic estimates suggest that COPD mortality rates will increase by 50% over the next 15 years. Individuals with COPD are at increased risk of cardiovascular diseases (CVD), lung cancer, osteoporosis and muscle wasting. Smoking is a well described risk factor for both COPD and CVD, but CVD in patients with COPD is likely to be due to other factors in addition to smoking. Systemic inflammation may be an important common etiological cause between COPD and CVD, being well described in both diseases. This paper reviews the close relationship between COPD and cardiovascular diseases, principally atherosclerosis. The common pathogenetic mechanisms, relation between cardiovascular comorbidities and pulmonary function parameters, the treatment of pulmonary and systemic inflammation, the role medications in the treatment of both disorders, the effect of cardiovascular comorbidities on the prognosis of COPD and prediction of mortality is discussed. The anti-inflammatory effects of inhaled corticosteroids and statins, their effects on cardiovascular endpoints, all-cause mortality, and survival of COPD patients are reviewed as a new perspective to the treatment. PMID- 19123088 TI - [Selective phosphodiesterase-4 (PDE-4) inhibitors in COPD]. AB - Chronic obstructive pulmonary disease (COPD) is a worldwide health problem resulting in significant morbidity and mortality; however, it could not be understood totally so far. Treatment options for the disease are quite limited and there is an urgent need for new treatment strategies. Among new therapeutic agents that are under development, a group of significant importance is phosphodiesterase-4 (PDE-4) inhibitors shown to have antiinflammatory actions. Phosphodiesterases are the enzymes responsible from the breakdown and inactivation of cyclic adenosine monophosphate (cAMP) which is an intracellular second messenger molecule. They are present in several structural and inflammatory cells, in these cells the inactivation of cAMP results in a proinflammatory cascade. So, in COPD which goes together with chronic inflammation, prevention of cAMP inactivation via phosphodiesterase enzyme inhibition made phosphodiesterase enzymes potential targets. Main phosphodiesterase playing a part in COPD is PDE-4 which is predominantly present in inflammatory cells and airway smooth muscle cells. The studies therefore focused on inhibitors selective to PDE-4 subtype. The two selective PDE-4 inhibitors that are at Phase III clinical trial stage are cilomilast and roflumilast. The studies have demonstrated that antiinflammatory effects of cilomilast and roflumilast positively contribute to the respiratory function, frequency of exacerbations and quality of life of COPD patients. Despite we need new studies to evaluate the influence of these agents on the natural course of COPD as well as their long-term safety; we can certainly comment that cilomilast and roflumilast are promising hope in COPD treatment by their clinical and antiinflammatory effects. PMID- 19123089 TI - Pleural metastasis from soft tissue sarcoma. PMID- 19123090 TI - Soft tissue sarcoma metastatic to pleura. PMID- 19123091 TI - Bacteria isolated from sewage influent resistant to ciprofloxacin, chloramphenicol and tetracycline. AB - This study assessed the presence of antibiotic-resistant bacteria in sewage influent. Resistance was measured by determining the lowest concentration of antibiotic, in micrograms per milliliter (microg mL(- 1)). To determine the minimum inhibitory concentration (MIC), which is used in diagnostic laboratories, we used the Etest, a plastic strip containing an antibiotic concentration gradient. In total, we sampled five sewage treatment plants of various sizes in Kansas and isolated bacteria resistant to three broad-spectrum antibiotics; ciprofloxacin (1-cyclopropyl-6-fluoro-4-oxo-7-piperazin-1-yl-quinoline-3 carboxylic acid), chloramphenicol 2,2-dichlor-N-[(aR, bR)-b-hydroxy-a hydroxymethyl-4-nitrophenethyl] acetamide), and tetracycline (2-(amino-hydroxy ethylidene)-4-dimethylamino-6,10,11,12a-tetrahydroxy-6-methyl-4,4a,5,5a tetrahydrotetracene-1,3,12-trione). In total, 25 Gram-negative isolates were found to be resistant to at least one of the antibiotics tested. Some isolates were multi-drug resistant, regardless of the amount of influent the sewage treatment plant received. A Pseudomonas isolate from the smallest sewage treatment plant (approximately 2 million gallons treated per day) showed resistance to all three antibiotics, albeit at low levels (10 microg mL(- 1)). The largest number of bacteria (6 species) were isolated from the largest sewage treatment plant (45 million gallons per day). Regardless, the results of this study are in agreement with similar studies, antibiotic resistance can persist long after the antibiotics have been forgotten. PMID- 19123092 TI - Biodegradation and detoxication of phenol by using free and immobilized cells of Acinetobacter sp. XA05 and Sphingomonas sp. FG03. AB - Strain XA05 and FG03 with high biodegradation activity of phenol were isolated from the activated sludge and phenol-contaminated soils in Northwest of China, respectively. DNA sequencing and homologous analysis of 16s rRNA gene identified that XA05 belonged to an Acinetobacter sp. and FG03 was closely related to the Sphingomonas sp. Cells of strain XA05 and FG03 were mixed at the ratio of 1:1, and polyvinyl alcohol (PVA) was used as a gel matrix to immobilize mixed cells by repeated freezing and thawing. Biodegradation was evaluated by determining phenol. Detoxication was evaluated by using Daphnia magna toxicity tests. The removal effciency of phenol and factors affecting phenol degradation were investigated, the stability of the immobilized cells was also reported. Experimental values indicated that both free cells and immobilized cells showed high phenol degradation effciencies, higher than 95% within 35 h with an initial concentration of 800 mg/L phenol, and the immobilized cells showed better performance than that of the suspended-culture cells. These results indicate that immobilized Acinetobncter sp. XA05 and Sphingomonas sp. FG03 possesses a good application potential in the treatment of phenol-containing wastewater. PMID- 19123093 TI - Changes of solid phase toxicity during sewage sludge composting in relation to bioavailability of polycyclic aromatic hydrocarbons. AB - The aim of the present study was to determine the content of the bioaccessible fraction of polycyclic aromatic hydrocarbons (PAHs) in sewage sludges and composts by means of three techniques (solid phase extraction with Tenax-TA and non-exhaustive extraction with hydroxypropyl[beta]cyclodextrin and n-butanol) and at the same time to establish their influence on toxicity for Heterocypris incongruens (Ostracodtoxkit test) and Lepidium sativum (Phytotoxkit test). In the majority of cases sewage sludges negatively influenced on organisms. Generally, sewage sludge composting exerted a positive influence on phytotoxicity, whereas in the case of ecotoxicity a negative effect was noted. The content of the potentially bioaccessible PAHs fraction varied depending on the method applied. Composting usually lowered the content of the potentially bioaccessible fraction. Significant positive relationships were observed mainly between Tenax-TA extracted fraction of individual PAHs and growth inhibition of H. incongruens. Only negative correlations were noted in the case of phytotoxicity. PMID- 19123094 TI - Kinetics of anaerobic degradation of screened dairy manure by upflow fixed bed digesters: effect of natural zeolite addition. AB - The effect of the hydraulic retention time (HRT) on the performance of two up flow anaerobic fixed bed digesters (UFAFBDs) packed with waste tire rubber (D1) and waste tire rubber and zeolite (D2) as micro-organism immobilization supports was studied. It was found that a first-order kinetic model described well the experimental results obtained. The kinetic constants for COD, BOD5, total solids (TS) and volatile solids (VS) removal were determined to be higher in digester D2 than in digester D1 or control. Specifically, they were 0.28 +/- 0.01, 0.32 +/- 0.02, 0.16 +/- 0.01 and 0.24 +/- 0.01 d(- 1) respectively for D1 and 0.33 +/- 0.02, 0.40 +/- 0.02, 0.21 +/- 0.01 and 0.28 +/- 0.01 d(- 1) respectively for D2. This was significant at the 95% confidence level. In addition, the first-order model was also adequate for assessing the effect of the HRT on the removal efficiency and methane production. Maximum methane yield and the first-order constant for methane production were determined and the results obtained were comparable with those obtained by other authors but operating at higher HRTs. Maximum methane yields and the kinetic constant for methane production were 11.1% and 29.4% higher in digester D2 than in D1. PMID- 19123095 TI - Iron oxide nanoparticle-assisted arsenic removal from aqueous system. AB - Iron oxide nanoparticles (IONPs) can play a significant role in the cycling of heavy metals. Arsenite [As(III)] is highly toxic, mobile, and predominant species in arsenic-contaminated groundwater. IONPs have been synthesized and tested for the removal of As(III) from arsenic contaminated water. In this work, we synthesized IONPs, as a finely divided loose nanopowder, using a chemical method involving a dispersion of the metal cations (Fe3+) through polymer molecules of polyvinyl alcohol (PVA) in an aqueous medium. X-ray diffraction (XRD) analysis confirmed the formation of a single phase rhombohedral crystal structure of R3c space group. Transmission electron microscopic images corroborate the result of IONPs of 45 nm average size and the rhombohedral shape. Selective experiments, conducted with an initial concentration of 0.25 ppm of As(III), have demonstrated the maximum As(III) adsorption capacity (96%) in 2.0 gL(- 1) IONPs in water at pH 4.5-7.5. At room temperature, the adsorption isotherm studies have revealed a better correlation with the Langmuir isotherm than the Freundlich isotherm. Characteristic surface hydrolysis of IONPs as = Fe-OH species has been studied in terms of the vibration bands. The results reveal that the removal of the As(III) species from water is associated with the As(III) adsorption onto the IONPs followed by a surface hydrolysis of the iron species. PMID- 19123096 TI - Enhanced arsenic removals through plant interactions in subsurface-flow constructed wetlands. AB - Arsenic (As) removal in pilot-scale subsurface-flow constructed wetlands (CWs) was investigated by comparing between CW units with vetiver grasses (CWplanted) and CW units without vetiver grasses (CWunplanted) in order to determine the roles of vetiver grasses affecting As removal. Based on the data obtained from 147 days of experiment, it is apparent that CWplanted units could remove As significantly higher than those of CWunplanted units with approximately 7-14%. Although analysis of As mass balance in CW units revealed that only 0.5-1.0% of total As was found in vetiver grasses, the As retained within bed of the CWplanted units (23.6-29.7 g) was higher than those in the CWunplanted units (21.3-26.8 g) at the end of the experiment, illustrating the effect of vetiver grasses on As accumulation in the CW units. Determination of As in different fractions in the CW bed suggested that the main mechanism of As retention was due mainly to As entrapment into the porous of bed materials (50-57% of total fraction), this mechanism is likely not affected by the presence of vetiver grasses. However, fraction of As-bound in organic matters that could be released from plant roots decomposition indicated the increase adsorption capacity of CW bed. In addition, organic sulfides produced from their root decomposition could help remove As through the precipitation/co-precipitation process. Under reducing condition in those CWplanted units, As could be leached out in the form of iron and manganese-bound complexes. PMID- 19123097 TI - Mixing of dissolved organic matter from distinct sources: using fluorescent pyrene as a probe. AB - Binary mixtures of humic substances (HSs) from three distinct sources (two soil humic acids and one natural organic matter) were used to investigate the mixing behavior of dissolved organic matters in the aquatic environment. Much longer time was required for equilibration of a solution of mixed HSs with pyrene as a probe, than for a solution comprising a single HS. Restructuring processes were responsible for the difference in equilibration kinetics and included conformational changes and aggregation. A major factor found controlling these processes was intermolecular interactions between components of HSs, although the concentration and hydrophobicity of the HSs were also important. In addition, the fluorescence quenching method yielded the binding constant (K DOC) of the HSs with pyrene. The K DOC of pyrene with binary humic mixtures was found to follow the conservative mixing rule. The findings of this study are applicable to studies of other hydrophobic organic pollutants and HSs and, will assist in modeling of the transport and fate of pollutants in aquatic environments. PMID- 19123098 TI - Photo-catalytic degradation of an oil-water emulsion using the photo-fenton treatment process: effects and statistical optimization. AB - The application of advanced oxidation processes (AOPs) to the treatment of an effluent contaminated with hydrocarbon oils was investigated. The AOPs conducted were Fe2+/H2O2 (Fenton's reagent), Fe2+/H2O2/UV (Photo-Fenton's reagent) and UV photolysis. These technologies utilize the very strong oxidizing power of hydroxyl radicals to oxidize organic compounds to harmless end products such as CO2 and H2O. A synthetic wastewater generated by emulsifying diesel oil and water was used. This wastewater might simulate, for example, a waste resulting from a hydrocarbon oil spill, onto which detergent was sprayed. The experiments utilising the Photo-Fenton treatment method with an artificial UV source, coupled with Fenton's reagent, suggest that the hydrocarbon oil is readily degradable, but that the emulsifying agent is much more resistant to degradation. The results showed that the COD (chemical oxygen demand) removal rate was affected by the Photo-Fenton parameters (Fe2+, H2O2 concentrations and the initial pH) of the aqueous solution. In addition, the applicability of the treatment method to a 'real' wastewater contaminated with hydrocarbon oil is demonstrated. The 'real' wastewater was sourced at a nearby car-wash facility located at a petroleum filling station and the experimental results demonstrate the effectiveness of the treatment method in this case. A statistical analysis of the experimental data using the Statistical Analysis System (SAS) and the response surface methodology (RSM) based on the experimental design was applied to optimize the Photo-Fenton parameters (concentrations of Fe2+, H2O2 and initial pH) and to maximize the COD removal rate (more than 70%). PMID- 19123099 TI - Effectiveness of removal of aqueous perchlorate by Cuprilig, a copper(II) derivative of Octolig. AB - The present study describes the effectiveness of removal of perchlorate ion by Octolig a commercially available immobilized ligand, IMLIG, and by Cuprilig, the cupric ion derivative of this material. Octolig consists of polyethylenediamine moieties covalently bound to a high-surface area silica gel (CAS Registry number = 404899-06-5). Perchlorate in drinking water is a nuisance anion that escapes from firms that produce rocket fuel, batteries, and fireworks and becomes a contaminant of groundwater. It may also be a natural component of the environment. Perchlorate ion can interfere with thyroid function and may also cause birth defects. Perchlorate contamination of water supplies is serious because of the difficulty in removing it, especially in the western United States where a combination of drought and water shortages exacerbate the problems of supplying safe drinking water to an increasing population. In this study, aqueous solutions were passed over a chromatography column containing samples of Cuprilig. Test water contained 70 microg perchlorate/L and effluent was below detection level (< 1 microg/L. Equally good results were obtained for Octolig in deionized water and in well water. Analyses were performed by a commercial laboratory. Mechanisms of interaction are proposed. PMID- 19123101 TI - Treatment of olive mill wastewater by a combined process: fenton's reagent and chemical coagulation. AB - Olive mill wastewater (OMW) was exposed to Fenton treatment and subjected to post Fenton coagulation/flocculation using lime. Several tests were performed for different R = H2O2/COD ratios, from 0.25 to 3.0, promoting the gradual addition of H2O2 and Fe2+, with the original condition of a pH = 3.5 and maintaining a constant relationship H2O2/Fe2+ = 15. A maximum reduction of COD of 96.8% was obtained using Fenton reagent with R = 3.0. In a second phase, the effluent which was neutralized with Ca(OH)2 reached a higher reduction of COD, achieving a final degradation value of 99.3%. Overall, the results show evidence of the potential value of applying the combined process tested. PMID- 19123100 TI - Comparing two empirical models used to predict process parameter dynamics during ozonation of synthetic dye wastewater. AB - Ozonation of a synthetic wastewater contaminated with acid yellow 17 dye and glucose was evaluated in a semi-batch bubble column. Data collected during this study suggests that the ozonation process removed color (represented as A400) more efficiently than chemical oxygen demand (COD) regardless of initial experimental conditions. With a 40-minute run time, the maximum A400 and COD removal efficiencies were 99.9 and 52.9%, respectively. Results indicated that the addition of COD to the system in the form of glucose had no effect on color removal efficiencies. The removal efficiencies of both color (A400) and COD in relation to the ozone utilized by the system were modeled using two previously derived empirical methods. Linear regression analysis was used to evaluate the effectiveness of both models to predict removal efficiencies of process parameters such as color and COD and to estimate ozone utilization. Results indicate both models predict color (A400) and COD removal as well as ozone utilization, but care should be taken when using the models to describe removal efficiencies when the wastewater inlet characteristics are variable. PMID- 19123102 TI - Leaching characteristics of selected South African fly ashes: effect of pH on the release of major and trace species. AB - Fly ash samples from two South African coal-fired power stations were subjected to different leaching tests under alkaline and acidic conditions in an attempt to assess the effect of pH on the leachability of species from the fly ashes and also assess the potential impact of the fly ashes disposal on groundwater and the receiving environment. To achieve this, German Standard leaching (DIN-S4) and Acid Neutralization Capacity (ANC) tests were employed. Mineralogical characterization of the fresh fly ashes revealed mullite and quartz as the major mineral phases with minor peaks of CaO and calcite. Chemical characterization by X-ray fluorescence (XRF) spectrometry revealed that the two fly ashes are similar, and consist of SiO(2), Al(2)O(3), Fe(2)O(3) and CaO as the main components with Cr, Co, Ni, Cu, Zn, V and Pb as minor components. Ca, Mg, Na, K and SO(4) were significantly leached into solution under the two leaching conditions with the total amounts in ANC leachates higher than that of DIN-S4. This indicates that a large fraction of the soluble salts in unweathered fly ash are easily leached. These species represents the fraction that can be flushed off initially from the surface of ash particles on contacting the ash with water. Al and Si were only observed in the leachates of the ANC test. Results obtained from the total acid-digestion and DIN-S4 leaching test indicated some toxic elements in the fly ashes are not easily solubilized. The amounts of toxic trace elements such as As, Se, Cd, Cr and Pb leached out of the fly ashes when in contact with de-mineralized water (DIN-S4 test) were low and below the Target Water Quality Range (TWQR) of South Africa. This is explained by their low concentrations in the fly ashes and their solubility dependence on the pH of the leaching solution. However the amounts of some minor elements such as B, Mn, Fe, As and Se leached out at lower pH ranging between 10 to 4 (ANC test) were slightly higher than the TWQR, an indication that the pH of the leaching solution plays a significant role on the leaching of species in fly ash. The high concentrations of the toxic elements released from the fly ashes at lower pH gives an indication that the disposal of the fly ash could have adverse effects on the receiving environment if the pH of the solution contacting the ashes is not properly monitored. The study indicated that on contact with water in a disposal scenario fly ash will release high amounts of soluble species. PMID- 19123103 TI - ICG fluorescence endoscope for visualization of the placental vascular network. AB - Intrauterine laser photocoagulation for twin-twin transfusion syndrome (TTTS) needs accurate in-situ recognition of placental vascular anastomosis. Because the conventional procedure is highly dependent upon the operators' skill and experience, we developed a new way to visualize the placental vascular network by a rigid-type fluorescence endoscope coupled with intravenous administration of Indocyanine green (ICG). The feasibility of the fluorescence endoscope was examined with monkey placentas and pregnant rats. The ICG fluorescence endoscope can visualize the placental vascular network in detail even in the presence of turbid amniotic fluid. Thus, this method is potentially useful for in-situ definition of the placental vascular anastomoses during the treatment for TTTS. In addition, our rigid-type fluorescence endoscope will also be a useful tool for lymph node dissection using ICG by endoscopic surgery. PMID- 19123104 TI - Cooling vest for improving surgeons' thermal comfort: a multidisciplinary design project. AB - A laparoscopic surgeon sometimes experiences heat-related discomfort even though the temperature situation is moderate. The aim of this project was to design a cooling vest using a phase change material to increase thermal comfort for the surgeon. The project focused on the design process to reveal the most important parameters for the design of a cooling vest that could be demonstrated in a clinical setting. We performed an entire design process, from problem analysis, situation observations, concept for a prototype, temperature measurements, and a final design based on clinical testing. The project was conducted by a multidisciplinary team consisting of product designers, engineers, physiologists, and surgeons. We carried out four physiological demonstrations of one surgeon's skin temperatures and heart rate during different laparoscopic procedures. A commercially available cooling vest for firemen and two proof-of-concept prototypes were tested alongside a reference operation without cooling. To aid the final design, one person went through a climate chamber test with two different set-ups of cooling elements. The final design was found to improve the conditions of our test subject. It was found that whole trunk cooling was more effective than only upper trunk cooling. A final design was proposed based on the design process and the findings in the operating room and in the laboratory. Although the experiences using the vest seemed positive, further studies on several operators and more surgical procedures are needed to determine the true benefits for the operator. PMID- 19123105 TI - Application of an interpersonal-psychological model of suicidal behavior to physicians and medical trainees. AB - Physicians and medical trainees (medical students and residents) are at increased risk for suicidal ideation. Yet few conceptual models have attempted to explain the elevated rates of suicide among physicians, and very little is known about what factors contribute to medical trainees' suicidal ideation and behaviors. In this paper, Joiner's (2005) interpersonal-psychological theory of suicidality will be explored as it applies to suicidal ideation and behavior among physicians and medical trainees. Literature addressing each component of the theory will be reviewed. Drawing upon extant data, each dimension of the theory (burden, thwarted belongingness, and acquired ability) will be examined in depth in terms of its applicability to suicidal thinking and behavior among physicians and physicians-in-training. Findings from the literature provide support for the interpersonal-psychological theory of suicidality as applied to this population. PMID- 19123106 TI - Bullying increased suicide risk: prospective study of Korean adolescents. AB - This study examines the independent impact of bullying on suicide risk. Bullying was assessed by peer nomination in a prospective study of 1,655 7th and 8th grade Korean students, and suicide by youth self-report. Odds Ratios (ORs) of bullying for suicidal risks were computed, controlling for other suicide risk factors. Victim-Perpetrators and female Victims at baseline showed increased risk for persistent suicidality (OR: 2.4-9.8). Male Incident Victims exhibited increased risk for suicidal behaviors and ideations (OR = 4.4, 3.6). Female Persistent Perpetrators exhibited increased risks for suicidal behaviors; male Incident Perpetrators had increased risk for suicidal ideations (OR = 2.7, 2.3). Baseline only male Victim-Perpetrators showed increased risk for suicidal ideations. (OR = 6.4). Bullying independently increased suicide risks. PMID- 19123107 TI - Personality style and impulsivity as determinants of suicidal subgroups. AB - Studies on suicide have often been inadequate in the sense of failing to understand or conceptualize the dynamics of suicidal behavior from a comprehensive theoretical basis. The aim of the present study was therefore to examine Blatt's theory of Dependent and Self-critical personality dimensions (Blatt, 1974, Blatt & Shichman, 1983) in the context of actual suicidal behavior, and also to utilize these dimensions to facilitate our understanding of suicidal action. Ninety-six undergraduate students who had attempted suicide in the past were administered a series of measures related to suicidal behavior, involving impulsivity, intent, and lethality. Participants were also identified as Dependent or Self-critical in their basic personality orientation. Dependents and Self-critics engaged in contrasting suicidal behaviors along several parameters, with Self-critical individuals generally posing greater risk. These differences were congruent with Blatt's theoretical framework. Implications for suicide risk assessment, management, and treatment are discussed, with special reference to the importance of identifying distinctive suicidal subgroups identified by Dependent and Self-critical personality dimensions. PMID- 19123108 TI - Characteristics of cocaine dependent patients who attempt suicide. AB - There have been few studies examining risk factors for attempting suicide among cocaine dependent patients. Therefore, a large group of 406 abstinent cocaine dependent patients had a psychiatric interview and completed the Childhood Trauma Questionnaire. Patients who had or had never attempted suicide were compared on putative suicide risk factors. The results showed that 43.5% of the patients had attempted suicide. Attempters were younger; more were female; more reported childhood trauma, a family history of suicidal behavior, a history of aggression, treatment with antidepressant medication, and comorbidity with alcohol and opiate dependence. Logistic regression revealed that childhood physical abuse, childhood emotional negelect, comorbidity with alcohol dependence, and treatment with antidepressant medication were significant predictors of attempting suicide. Thus these results suggest that attempting suicide is common among cocaine dependent patients and that both distal and proximal risk factors may play a role. PMID- 19123109 TI - Suicide prevention by online support groups: an action theory-based model of emotional first aid. AB - In the last two decades, online support groups have become a valuable source of help for individuals in suicidal crisis. Their attractiveness is attributed to features that enhance help-seeking and self-disclosure such as availability, anonymity, and use of written communication. However, online support groups also suffer from limitations and potential risks as agents of suicide prevention. The Israeli Association for Emotional First Aid (ERAN) has developed a practical model that seeks to maximize the benefits and minimize the risks of online suicide prevention. The model applies the Action Theory concepts whereby individuals shape their own environment. The present paper presents the model, which is based on an online support group combined with personal chat and a telephonic help line. The online support group is moderated by paraprofessionals who function as both process regulators and support providers. The principles and practice of the model are described, the theoretical rationale is presented, and directions for future research are suggested. PMID- 19123110 TI - Psychopathic traits and suicidal ideation in high-school students. AB - The aim of the study was to evaluate the contribution of psychopathic traits in the prediction of suicidal ideation among non-clinical non-forensic adolescents. A sample of 312 high-school students completed questionnaires assessing suicidal ideation, depressive symptoms, borderline personality traits, and cannabis use and the Youth Psychopathic traits Inventory (YPI, Andershed, Kerr, Stattin et al., 2002), which consists of three dimensions, the Interpersonal subscale (grandiosity, manipulation), the Affective subscale (callousness, unemotionality), and the Behavioral subscale (impulsiveness, irresponsibility). A multiple regression analysis showed that the affective component of psychopathic traits was an independent predictor of suicidal ideation. Our results suggest that clinicians should not assume that the presence of psychopathic traits in adolescent is a protection against suicidal ideation. PMID- 19123111 TI - Defense mechanisms and suicide risk in major depression. AB - Past suicide attempt has been consistently reported to be the best predictor of future suicide attempt. The need to identify predictors that can be targets of therapy is of great importance. Coping styles and defense mechanisms have now been linked to suicide in numerous reports. In this study, we expand on past research by exploring differences in defense mechanism use between three groups: 1) without current suicide ideation/attempt; 2) with current suicide ideation/no attempt; and 3) with current suicide attempt. We also explored the contribution of covariates, such as symptom severity and past attempt. Seventy-five adult patients who were within 48 hours of hospital admission for current major depressive episode were recruited. Clinical interview was conducted to verify diagnosis and assess symptom severity. Patients completed the Defense Style Questionnaire within 48 hours of admission based on their current thoughts and beliefs. Logistic regressions were used to determine the best predictors of current suicide status. Consistently, the use of more image-distorting mechanisms was the best predictor of current suicide attempt. Symptom severity and past attempt were not significant predictors after controlling for influence of defense styles. Decreased use of image-distorting mechanisms in adult patients with major depressive disorder should be considered as a distinct target of therapy. PMID- 19123112 TI - Policy-appropriate measurement of suicide: headcount vs. potential years of life lost, Australia, 1907-2005. AB - We reconsider conventional suicide measurement. First, a headcount of suicide is examined relative to some other causes of death (circulatory diseases, cancer, and motor vehicle accidents). We then construct a time-series data set of an alternative measure of suicide, the potential years of life lost (PYLL) for males and females. Suicide PYLLs average 4.57% of all male PYLLs and 2.44% of female PYLLs for 1907-2005. The comparable "count" percentages are 1.85 and 0.65, respectively. These differences are widening through time. In 2005, suicide represented 3.25% of all male deaths and 0.90% of female deaths using the count measure and, using PYLLs, 11.0% and 4.96%, respectively. The two measures produce quite different indications of suicide. PMID- 19123113 TI - A psychological autopsy study of suicidal hanging from Cuttack, India: focus on stressful life situations. AB - Factors and stressful life situations associated with suicidal hanging in a sample from India were studied with a view to explore preventability. Information was collected on consecutive suicidal hanging victims in a 2-year period from various sources including family members through psychological autopsy method. Out of 662 autopsies involving suicide during the study period, 104 had used hanging as the method. Age group of 21-30 years, married females, unmarried males, dowry related stress, unemployment, prolonged illness, failure in examinations, relationship and financial problems were associated more frequently with suicidal hanging. Stresses stemming from social practices and perceptions are linked with considerable number of suicidal hanging, which suggest priority areas for intervention. PMID- 19123114 TI - An exploratory study of linguistic-colour associations across languages in multilingual synaesthetes. AB - In one of the most common forms of synaesthesia, linguistic-colour synaesthesia, colour is induced by stimuli such as numbers, letters, days of the week, and months of the year. It is not clear, however, whether linguistic-colour synaesthesia is determined more by higher level semantic information--that is, word meaning--or by lower level grapheme or phoneme structure. To explore this issue, we tested whether colour is consistently induced by grapheme or phoneme form or word meaning in bilingual and trilingual linguistic-colour synaesthetes. We reasoned that if the induced colour was related to word meaning, rather than to the acoustic or visual properties of the words, then the induced colours would remain consistent across languages. We found that colours were not consistently related to word meaning across languages. Instead, induced colours were more related to form properties of the word across languages, particularly visual structure. However, the type of inducing stimulus influenced specific colour associations. For example, colours to months of the year were more consistent across languages than were colours to numbers or days of the week. Furthermore, the effect of inducing stimuli was also associated with the age of acquisition of additional languages. Our findings are discussed with reference to a critical period in language acquisition on synaesthesia. PMID- 19123115 TI - Training with anxiety has a positive effect on expert perceptual-motor performance under pressure. AB - In two experiments, we examined whether training with anxiety can prevent choking in experts performing perceptual-motor tasks. In Experiment 1, 17 expert basketball players practised free throws over a 5-week period with or without induced anxiety. Only after training with anxiety did performance no longer deteriorate during the anxiety posttest. In Experiment 2, 17 expert dart players practised dart throwing from a position high or low on a climbing wall, thus with or without anxiety. Again, only after training with anxiety was performance maintained during the anxiety posttest, despite higher levels of anxiety, heart rate, and perceived effort. It is concluded that practising under anxiety can prevent choking in expert perceptual-motor performance, as one acclimatizes to the specific processes accompanying anxiety. PMID- 19123116 TI - Name-picture verification as a control measure for object naming: a task analysis and norms for a large set of pictures. AB - The name-picture verification task is widely used in spoken production studies to control for nonlexical differences between picture sets. In this task a word is presented first and followed, after a pause, by a picture. Participants must then make a speeded decision on whether both word and picture refer to the same object. Using regression analyses, we systematically explored the characteristics of this task by assessing the independent contribution of a series of factors that have been found relevant for picture naming in previous studies. We found that, for "match" responses, both visual and conceptual factors played a role, but lexical variables were not significant contributors. No clear pattern emerged from the analysis of "no-match" responses. We interpret these results as validating the use of "match" latencies as control variables in studies or spoken production using picture naming. Norms for match and no-match responses for 396 line drawings taken from Cycowicz, Friedman, Rothstein, and Snodgrass (1997) can be downloaded at: http://language.psy.bris.ac.uk/name-picture_verification.html. PMID- 19123117 TI - No evidence for a cue mismatch in negative priming. AB - An experiment is reported in which the cue mismatch hypothesis of negative priming, an important novel variant of the mismatching hypothesis, was tested. A cue mismatch and a no mismatch condition were contrasted in a visual discrimination task. In the prime display of cue mismatch ignored-repetition trials, the colour of the prime distractor was different from the colour of the cue indicating the selection feature (coloured square). In probe displays, cue and repeated stimulus had the same colour. In the no mismatch condition, the visual cue was neutral in terms of colour (always black), so that there was always no cue mismatch between prime and probe displays. Contrary to the prediction of the cue mismatch hypothesis, the negative priming effect was not larger in the cue mismatch than in the no mismatch condition. The cue mismatch hypothesis must therefore be rejected. In contrast, the episodic retrieval account is consistent with the results. PMID- 19123118 TI - Working-memory capacity predicts the executive control of visual search among distractors: the influences of sustained and selective attention. AB - Variation in working-memory capacity (WMC) predicts individual differences in only some attention-control capabilities. Whereas higher WMC subjects outperform lower WMC subjects in tasks requiring the restraint of prepotent but inappropriate responses, and the constraint of attentional focus to target stimuli against distractors, they do not differ in prototypical visual-search tasks, even those that yield steep search slopes and engender top-down control. The present three experiments tested whether WMC, as measured by complex memory span tasks, would predict search latencies when the 1-8 target locations to be searched appeared alone, versus appearing among distractor locations to be ignored, with the latter requiring selective attentional focus. Subjects viewed target-location cues and then fixated on those locations over either long (1,500 1,550 ms) or short (300 ms) delays. Higher WMC subjects identified targets faster than did lower WMC subjects only in the presence of distractors and only over long fixation delays. WMC thus appears to affect subjects' ability to maintain a constrained attentional focus over time. PMID- 19123119 TI - Rational numbers: componential versus holistic representation of fractions in a magnitude comparison task. AB - This study investigated whether the mental representation of the fraction magnitude was componential and/or holistic in a numerical comparison task performed by adults. In Experiment 1, the comparison of fractions with common numerators (x/a_x/b) and of fractions with common denominators (a/x_b/x) primed the comparison of natural numbers. In Experiment 2, fillers (i.e., fractions without common components) were added to reduce the regularity of the stimuli. In both experiments, distance effects indicated that participants compared the numerators for a/x_b/x fractions, but that the magnitudes of the whole fractions were accessed and compared for x/a_x/b fractions. The priming effect of x/a_x/b fractions on natural numbers suggested that the interference of the denominator magnitude was controlled during the comparison of these fractions. These results suggested a hybrid representation of their magnitude (i.e., componential and holistic). In conclusion, the magnitude of the whole fraction can be accessed, probably by estimating the ratio between the magnitude of the denominator and the magnitude of the numerator. However, adults might prefer to rely on the magnitudes of the components and compare the magnitudes of the whole fractions only when the use of a componential strategy is made difficult. PMID- 19123120 TI - Early and long-term evaluation of perfusion changes in free DIEP-flaps for breast reconstruction via IC-view and duplex ultrasound: autonomous or peripheral perfusion? AB - Controversy surrounds the question whether free flaps remain dependent for blood supply on their vascular pedicle or if there is an autonomization by neovascularization from the surrounding wound bed. This becomes important when flap shaping or repositioning are performed. Our study involved 23 patients who received a deep inferior epigastric perforator (DIEP) flap for breast reconstruction. IC-View laser video angiography (Pulsion Medical Systems AG, Munich, Germany) was executed immediately and 18 months postoperatively. Two zones (close and distant to the pedicle) and the contralateral breast were compared quantitatively. Via duplex ultrasound, late perfusion changes were measured to analyze flow volume, velocity, and diameter of the internal thoracic artery and the DIEP-flap pedicle. In the long term, early postoperative flap hyperperfusion changed to flap hypoperfusion. No enhanced flow from the flap surrounding into the flap borders was measured. These results might indicate a long-term increase in total peripheral vascular resistance of the transplanted tissue. Postoperative perfusion after 18 months remains dependent on the anastomosed vascular pedicle. PMID- 19123121 TI - Tissue plug technique for management of large chronic empyema defects and bronchopleural fistulas. AB - After multiple thoracotomies, local muscles are often transected or sacrificed for the treatment of intrathoracic infection. In these conditions, free tissue transfers are described as an alternative. Five anterolateral thigh musculocutaneous free flaps were used for the treatment of complicated large chronic empyema defects and bronchopleural fistulas with a special flap inset and gauze-packing method, the so-called tissue plug technique. All flaps survived completely without complication. There was no sign of recurrent infection, and the bronchopleural fistulas were sealed and the empyema cavities completely obliterated. We concluded that the tissue plug technique is an alternative for the management of difficult empyema cavities and bronchopleural fistulas in selected patients where the soft tissue flap is not enough to fill the complete dead spaces. PMID- 19123122 TI - Neural stem cells at the crossroads: MMPs may tell the way. AB - Matrix metalloproteinases (MMP) constitute a family of more than 25 enzymes which process a large number of pericellular substrates. Even though initially reported to have an ability to degrade almost all of the extracellular components, MMP are now known to play roles which are not limited to the breakdown of extracellular barriers. In fact, MMPs regulate many biological processes, being involved not only in physiological events, but also in pathological processes. Strikingly, MMPs have been found to be involved in the physiology of the Central Nervous System (CNS), taking part and playing important roles in several processes such as repair and ontogeny, as well as in pathological conditions of the CNS. Initially considered to be a static structure, lacking regenerative capability, the CNS has been considered for a long time to be a system without renewal capabilities. Recently, the discovery of constant neural replacement has changed our way of considering the adult brain, and the finding of the existence of neural stem cells has opened the way to exciting and fascinating perspectives of the CNS. So, could MMPs, originally found during metamorphosis in tadpoles, and now amazingly identified in the CNS, have something to do in neuronal function? In this review we take into consideration the possible roles of two metalloproteinases, MMP-2 and MMP-9, also called gelatinases, in controlling several aspects of CNS organization, including the modulation of neural stem cell properties and the differentiation of their progeny, both under normal and pathophysiological conditions. PMID- 19123123 TI - Organelles in developing neurons: essential regulators of neuronal morphogenesis and function. AB - Eukaryotic cells contain multiple intracellular organelles which are structurally and functionally distinct membrane-delimited compartments. Organelles play vital roles in many cellular events in essentially all eukaryotic cells. Although the canonical roles of organelles are well described by classical in vitro studies, little is known about the specific physiological roles of organelles in neurons, which possess extremely polarized cellular structures and have a massive cellular volume compared with most eukaryotic cells. Studies that make use of recently developed genetic and microscopic techniques are currently elucidating the unexpectedly specialized roles of intracellular, membrane-delimited organelles in neuronal morphogenesis and function, and in human disease. Here we review recent advances in understanding the roles of organelles (the ER-Golgi secretory pathway, endosomes and mitochondria) in developing neurons. PMID- 19123124 TI - Ocular forkhead transcription factors: seeing eye to eye. AB - Forkhead transcription factors comprise a large family of proteins with diverse functions during development. Recently, there has been accumulating evidence that several members of this family of proteins play an important role in the development of the vertebrate retina. Here, we summarize the cumulative data which demonstrates the integral role that forkhead factors play in cell cycle control of retinal precursors, as well as in cell fate determination, during retinal development. The expression patterns for 14 retinal expressed forkhead transcription factors are presented with an emphasis on comparing the expression profiles across species. The functional data regarding forkhead gene products expressed within the retina are discussed. As presented, these data suggest that forkhead gene products contribute to the complex regulation of proliferation and differentiation of retinal precursors during vertebrate eye development. PMID- 19123125 TI - Loss of REEP4 causes paralysis of the Xenopus embryo. AB - Members of the REEP (Receptor expression enhancing protein) family contain a TB2/DP1, HVA22 domain that is involved in intracellular trafficking and secretion. Consistent with the presence of this domain, REEP1 and REEP3 enhance the expression of odorant and taste receptors in mammals, while mutation of these genes causes defects in neural development. REEP4 was identified in the course of a functional antisense morpholino oligonucleotide screen searching for genes involved in the early development of Xenopus tropicalis: although over-expression of the gene causes no phenotype, embryos lacking REEP4 develop a slightly kinked body axis and are paralysed. At tailbud stages of development, REEP4 is expressed in the somites and neural tube. The paralysis observed in embryos lacking REEP4 might therefore be caused by defects in the nervous system or in muscle. To address this point, we examined the expression of various neural and muscle markers and found that although all are expressed normally at early stages of development, many are down regulated by the tailbud stage. This suggests that REEP4 plays a role in the maintenance of both the nervous system and the musculature. PMID- 19123126 TI - Hedgehog signalling is required for cloacal development in the zebrafish embryo. AB - The Hedgehog (Hh) family of signalling molecules is essential for a wide range of developmental processes. Mammalian studies have implicated the Hedgehog pathway in the aetiology of anorectal malformations (ARMs), relatively common congenital anomalies caused by failures in the development of the cloaca. In this study we demonstrate that Hh signalling is absolutely required for the formation of the zebrafish cloaca and that the severity of the posterior gut abnormalities induced by a reduction in Hh activity is dependent on the levels of Hh signal transduction. The complete loss of all Hh activity results in the most severe defects and the critical period for Hh activity is between 34 and 74 hours post fertilisation. Using a range of mutant genotypes that cause notochord and floorplate abnormalities, we show that the source of the Hh signals required for posterior gut formation is the endoderm and not the notochord, as previously postulated in mammalian models of ARMs. We show that Adriamycin, a drug known to cause ARMs in rat, but not chick embryos, has no effect on the development of the zebrafish gastrointestinal tract. These studies establish the zebrafish as a model for ARMs, and for the elucidation of other pathways involved in hindgut developmental processes. PMID- 19123127 TI - Genetic evidence against a role for W-linked histidine triad nucleotide binding protein (HINTW) in avian sex determination. AB - Birds have a ZZ/ZW sex chromosome system, but the mechanism of sex determination remains unknown. The heterogametic sex is female (ZW) and one hypothesis holds that the W chromosome carries a dominant-acting ovary-determining gene. The strongest candidate ovary-determinant on the W chromosome is HINTW, which encodes an aberrant nucleotide hydrolase enzyme. HINTW is conserved amongst all carinate (flying) birds and it is strongly expressed in the gonads and other tissues of female chicken embryos. This and other lines of circumstantial evidence support the proposal that HINTW is the female-determining gene in birds. However, in vivo gain-of-function or loss-of-function studies have not hitherto been reported to test this hypothesis. We tested the potential role of HINTW by mis-expressing it in genetically male (ZZ) embryos, using the RCASBP avian retroviral vector. Strong, widespread expression was delivered throughout the embryo, including the urogenital system, as assessed by whole mount in situ hybridisation. This expression pattern mimicked that seen in normal ZW females, in which HINTW is widely expressed. Strong mis-expression was observed throughout the gonads of genetic male (ZZ) embryos. However, despite strong HINTW expression, ZZ gonads developed normally as bilateral testes. In tissue sections of ZZ urogenital systems transgenic for HINTW, normal testicular histology was observed. Female (ZW) gonads over-expressing HINTW also developed normally, with normal ovarian structure and left/right asymmetry. These results provide genetic evidence against a dominant role for HINTW in avian sex determination. PMID- 19123128 TI - Transcriptional regulation by Pax3 and TGFbeta2 signaling: a potential gene regulatory network in neural crest development. AB - Pax3 regulates neural crest cell migration and is critical during neural crest development. TGFbs modify neural crest cell migration and differentiation. TGFbeta2 nullizygous embryos (TGFbeta2(-/-)Pax3(+/+)) display open neural tube and bifid spine, whereas in wild type embryos, the neural tube is closed. In previous work, we have demonstrated that Pax3 regulates TGFbeta2 by directly binding to cis-regulatory elements on its promoter. In this study, we found that the TGFbeta2 nullizygous phenotype can be reversed to the wild type phenotype by down-regulating one allele of Pax3, as in TGFbeta2(-/-)Pax3(+/-) embryos obtained through breeding TGFb2(+/-)Pax3(+/-) mice. The data in this paper suggest that Pax3 and TGFbeta2 interact in a coordinated gene regulatory network, linked by common downstream effector genes, to bring about this phenotypic reversal. Downstream effectors may include Hes1, Ngn2 and Sox9, as well as other genes involved in neuronal differentiation. PMID- 19123129 TI - Lef1 plays a role in patterning the mesoderm and ectoderm in Xenopus tropicalis. AB - Tcf/Lef HMG box transcription factors are nuclear effectors of the canonical Wnt signaling pathway, which function in cell fate specification. Lef1 is required for the development of tissues and organs that depend on epithelial mesenchymal interactions. Here, we report the effects of lef1 loss of function on early development in X. tropicalis. Depletion of lef1 affects gene expression already during gastrulation and results in abnormal differentiation of cells derived from ectoderm and mesoderm. At tail bud stages, the epidermis was devoid of ciliated cells and derivatives of the neural crest, e.g. melanocytes and cephalic ganglia were absent. In the Central Nervous System, nerve fibers were absent or underdeveloped. The development of the paraxial mesoderm was affected; intersomitic boundaries were not distinct and development of the hypaxial musculature was impaired. The development of the pronephros and pronephric ducts was disturbed. Most striking was the absence of blood flow in lef1 depleted embryos. Analysis of blood vessel marker genes demonstrated that lef1 is required for the development of the major blood vessels and the heart. PMID- 19123130 TI - The heart forming region of early chick embryo is an alternative source of embryonic stem cells. AB - In early chick embryo, the precardiac cells reside within distinct groups of mesodermal cells known as presumptive heart forming regions (HFRs). HFRs are located on the lateral sides of the Hensens node. In an effort to study fate of HFRs in isolation, HFRs were excised from early gastrulating chick embryos and cultured in vitro. A very small proportion of HFRs from 18 h incubated embryos differentiated into beating cardiomyocytes whereas about 43% of HFRs from embryos incubated for longer durations (20, 23 and 28 h) showed beating activity. The potential of HFRs, from 18 h incubated embryos, to differentiate into cardiomyocytes increased significantly in presence of Hensens node. About one third of the HFR cells underwent spontaneous differentiation into adipocytes in culture. Simultaneously, some of the cells derived from HFRs exhibited alkaline phosphatase (AP) activity indicating presence of stem cells in the culture. HFR cells were positive for vimentin indicating their mesenchymal origin. FGF supplement increased the proportion of AP-positive cells in a dose dependent manner. The present study demonstrates that HFRs can serve as a source of mesenchymal stem cells which can be gainfully employed for various purposes. The results also suggest that even though the in vitro cultured HFRs from 18 h incubated HH stage 4 chick embryo retain the potential to undergo cardiac differentiation, certain instructive signals from Hensens node may reinforce the fate. PMID- 19123131 TI - Triiodothyronine (T3) action on aquatic locomotor behavior during metamorphosis of the bullfrog Rana catesbeiana. AB - Thyroid hormones--particularly triiodothyronine, T3--play a critical role in the morphological transformations comprising metamorphosis in larval bullfrogs (Rana catesbeiana). Traditional staging criteria for anuran larvae incompletely distinguish physiological and behavioral changes during growth. We therefore first developed a new parameter to describe larval growth, the developmental index (DI), which is simply the ratio between the tail length of the larva and its head diameter. Using the DI we were able to identify two distinct populations classifying the larvae during growth along a continuous linear scale with a cutoff value of DI at 2.8. Classification based on the DI, used in this study, proved an effective complement to existing classifications based on developmental staging into pre- or pro-metamorphic stages. Exposure to T3 in the water induced a rapid (beginning within 5 min) and significant decrease (approximately 20-40%) in locomotor activity, measured as total distance traversed and velocity. The largest decrease occurred in more developed larvae (DI<2.8). To determine correlated changes in the neuromuscular junctions during metamorphosis and apoptotic tail loss, miniature endplate currents from tail muscle were recorded during acute exposure to a hypertonic solution, which simulates an apoptotic volume decrease. Our results support a role for T3 in regulating larval locomotor activity during development, and suggest an enhanced response to volume depletion at the neuromuscular junction of older larvae (DI<2.8) compared to younger animals (DI> or =2.8). We discuss the significance of the possible role of an apoptotic volume decrease at the level of the neuromuscular junction. PMID- 19123132 TI - Ceratitis capitata transformer-2 gene is required to establish and maintain the autoregulation of Cctra, the master gene for female sex determination. AB - In Drosophila melanogaster, transformer-2 (TRA-2) which is a non-sex-specific auxiliary splicing factor, is required to promote female sexual differentiation by interaction with the female-specific TRA. The two proteins positively regulate the splicing of both doublesex (dsx) and fruitless (fru) pre-mRNAs, which in turn regulate phenotypic and behavioural sexual dimorphism. In the Mediterranean fruitfly Ceratitis capitata, the female-specific CcTRA is similarly required not only for Ccdsx splicing, but also to exert a novel autoregulatory function that consists of promoting female-specific splicing of Cctra pre-mRNA. This study reports the isolation and functional analysis of the C. capitata homologue of the Drosophila transformer-2 gene (Cctra-2). Transient RNAi against Cctra-2 during embryonic development causes the full sex reversal of XX flies in adult fertile pseudo-males, as well as changes in the splicing pattern of Cctra, Ccdsx and Ccfruitless (Ccfru). We propose that: 1) Cctra-2, as in Drosophila, is necessary for promoting Ccdsx and putative Ccfru pre-mRNA female-specific splicing and that 2) unlike in Drosophila, Cctra-2 appears to be necessary for establishing female sex determination in early XX embryos and for maintaining the positive feedback regulation of Cctra during development. PMID- 19123133 TI - Acquisition of plastid movement responsiveness to light during mesophyll cell differentiation. AB - A culture of Nicotiana tabacum leaf protoplasts, regenerating in vitro, was used to study light-induced plastid translocations. Experiments were carried out for 5 months starting with protoplasts, through single cells, microcolonies and callus to the differentiated mesophyll of regenerated plants. Although the actin cytoskeleton was fully developed at every stage of culture, blue light-mediated directional movements of chloroplasts were observed only after the full differentiation of callus into leaf tissues. These chloroplast rearrangements were similar to those observed in control plants grown from seeds. Under strong blue light, chloroplasts gathered at the cell walls parallel to the light direction (profile position); under weak blue light, they gathered at the walls perpendicular to the light direction (face position). No light-dependent plastid arrangements were found in undifferentiated cell cultures even after cell wall recovery. A characteristic pattern of plastids in the dividing cells was independent of light signals. Only trace chloroplast responses to strong blue light were detected in the first leaves regenerating from callus. We hypothesize that factors which control the developmental status of a cell, i.e. division and differentiation, take over the control of plastid redistribution from light signals. PMID- 19123134 TI - The 2-cell block occurring during development of outbred mouse embryos is rescued by cytoplasmic factors present in inbred metaphase II oocytes. AB - In mice, completion of preimplantation development in vitro is restricted to certain crosses between inbred strains. Most of the outbred and inbred strains cease development at the 2-cell stage, a phenomenon known as the "2-cell block". Reciprocal mating between blocking and non-blocking strains has shown that the 2 cell block is dependent upon female, but not male, developmental information. One question that still remains unanswered is whether the genome of the metaphase II (MII) oocyte is genetically programmed to express, during the very early stages of development, some factor(s) required to determine developmental competence beyond the 2-cell stage. In the present study, we have addressed this question by performing reciprocal MII-chromosome plate transfer between MII oocytes of a non blocking inbred strain and MII oocytes of a blocking outbred strain. Here, we report that development beyond the 2-cell stage does not depend on the MII genome, but instead it relies on a cytoplasmic factor(s) already present in ovulated non-blocking oocytes, but absent, inactive or quantitatively insufficient in blocking oocytes. Further evidence of the ooplasmic origin of this component(s) was obtained by transferring a small quantity of ooplasm from non-blocking MII oocytes to blocking MII oocytes or 2-cell embryos. Following the transfer, a high percentage of blocking oocytes/embryos acquired developmental competence beyond the 2-cell stage and reached the blastocyst stage. This study shows that development beyond the 2-cell stage relies also on a factor(s) already present in the ovulated oocyte. PMID- 19123135 TI - Rotation in Xenopus laevis embryos during the second cell cycle. AB - Using time-lapse video recording and comparing successive digital images, we found that 38% of Xenopus laevis embryos (n=118) exhibited rotation during the second cell cycle. This rotation, which we term the second rotation, started approximately during the appearance of the first cleavage furrow and proceeded clockwise or counterclockwise around the vertical axis. Rotations lasted for 5-30 minutes, i.e. up to the beginning of the third cell cycle. The mean rotation angle was 36.4 degrees, with a maximum rotation of 77 degrees. No mortality was observed among the embryos exhibiting rotation. The second rotation was observed to be similar to the well-known fertilization rotation which takes place during the first cell cycle. The possible nature and significance of the second rotation are discussed. PMID- 19123136 TI - Generation of a reporter-null allele of Ppap2b/Lpp3and its expression during embryogenesis. AB - Our knowledge of how bioactive lipids participate during development has been limited principally due to the difficulties of working with lipids. The availability of some of these lipids is regulated by the Lipid phosphate phosphatases (LPPs). The targeted inactivation of Ppap2b, which codes for the isoenzyme Lpp3, has profound developmental defects. Lpp3 deficient embryos die around E9.5 due to extraembryonic vascular defects, making difficult to analyze its participation in later stages of mouse development. To gain some predictive information regarding the possible participation of Lpp3 in later stages of development, we generated a Ppap2b null reporter allele and it was used to establish its expression pattern in E8.5-13.5 embryos. We found that Ppap2b expression during these stages was highly dynamic with significant expression in structures where multiple inductive interactions occur such as the limb buds, mammary gland primordia, heart cushions and valves among others. These observations suggest that Lpp3 expression may play a key role in modulating/integrating multiple signaling pathways during development. PMID- 19123137 TI - Sim1 and Sim2 expression during chick and mouse limb development. AB - The Drosophila Single minded (Sim) transcription factor is a master regulator of cell fate during midline development. The homolog mouse Sim1 and Sim2 genes are important for central nervous system development. Loss of mSim1 activity leads to an absence of specific neuroendocrine lineages within the hypothalamus, while overexpression of mSim2 leads to behavioural defects. We now provide evidence that vertebrate Sim genes might be important for limb muscle formation. We have examined by in situ hybridisation the expression of the Sim1 and Sim2 genes during limb development in chick and mouse embryos. The expression of both Sim genes is mainly associated with limb muscle formation. We found that each Sim gene has a similar temporal and spatial expression pattern in chick and mouse embryonic limbs, although with some differences for the Sim2 gene between species. In chick or mouse embryonic limbs, Sim1 and Sim2 display non-overlapping expression domains, suggesting an involvement for Sim1 and Sim2 proteins at different steps of limb muscle formation. Sim1 gene expression is associated with the early step of muscle progenitor cell migration in chick and mouse, while the Sim2 gene is expressed just after the migration process. In addition, chick and mouse Sim2 gene expression is enhanced in limb ventral muscle masses versus dorsal ventral muscle masses. Our results provide a basis for further functional analysis of the Sim genes in limb muscle formation. PMID- 19123138 TI - Expression of Xenopus tropicalis HNF6/Onecut-1. AB - Onecut genes belong to a family of transcription factors that are known to be important in embryonic development. In the present study, we analyzed the pattern of expression of Onecut-1/HNF6 in Xenopus tropicalis using RT-PCR and whole mount in situ hybridization. Expression of the Xenopus tropicalis Onecut-1/HNF6 gene was found to be conserved in the neural tube, the sensory placodes and in the anterior ventral endoderm in a domain consistent with the developing liver primordium. PMID- 19123139 TI - Expression patterns of Src-family tyrosine kinases during Xenopus laevis development. AB - Src family tyrosine kinases (SFKs) play important roles in cell morphology, differentiation, motility and proliferation. Elevated expression and/or specific activity of Src kinases are characteristic for several types of human cancer. However, little information is available about the role and spatio-temporal expression of SFKs in early embryonic development. In this study we characterized, in Xenopus laevis, the expression patterns of five SFK genes src, fyn, yes, lyn and laloo as well as of the csk gene, a negative regulator of SFKs, using RT-qPCR and in situ hybridisation. We found that transcripts of all SFKs and csk were already detectable in one-cell embryos and their levels similarly oscillated during subsequent development. First, after stage 8, the levels of SFK and csk mRNAs began to decrease, reached a minimum between stages 10 and 28 and increased again. In the later stages (33-45), the levels of fyn, yes and csk mRNAs returned to approximately maternal ones, whereas the src, laloo and lyn mRNA transcripts exceeded, up to about 3.5-6-fold, their maternal levels. In situ hybridisation analysis located the SFK and csk transcripts in the animal hemisphere of Xenopus embryos. Subsequent gastrula stages showed signals in ectodermal cells, mid-neurula stage embryos at neural folds, and the tailbud stages showed strong signals in the brain and neural tube. RT-qPCR concentration profiling along the animal-vegetal axis proved in blastula and gastrula the preferential localisation of yes, src, lyn and csk transcripts towards the animal pole in a gradient-like manner. In contrast, laloo and fyn displayed a vegetal pole preference. PMID- 19123140 TI - Developmental expression of Apnanos during oogenesis and embryogenesis in the parthenogenetic pea aphid Acyrthosiphon pisum. AB - Among genes that are preferentially expressed in germ cells, nanos and vasa are the two most conserved germline markers in animals. Both genes are usually expressed in germ cells in the adult gonads, and often also during embryogenesis. Both nanos-first or vasa-first expression patterns have been observed in embryos, implying that the molecular networks governing germline development vary among species. Previously we identified Apvasa, a vasa homologue expressed in germ cells throughout all developmental stages in the parthenogenetic and viviparous pea aphid Acyrthosiphon pisum. In asexual A. pisum, oogenesis is followed by embryogenesis, and both occur within the ovarioles. In order to understand the temporal and spatial distribution of nanos versus vasa during oogenesis and embryogenesis, we isolated a nanos homologue, Apnanos, and studied its expression. In adults, Apnanos is preferentially expressed in the ovaries. In early embryos, Apnanos transcripts are localized to the cytoplasm of cellularizing germ cells, and soon thereafter are restricted to the newly segregated germ cells in the posterior region of the cellularized blastoderm. These results strongly suggest that the Apnanos gene is a germline marker and is involved in germline specification in asexual A. pisum. However, during the middle stages of development, when germline migration occurs, Apnanos is not expressed in the migrating germ cells expressing Apvasa, suggesting that Apnanos is not directly associated with germline migration. PMID- 19123141 TI - Arabidopsis monomeric G-proteins, markers of early and late events in cell differentiation. AB - In Schizosaccharomyces pombe, septum formation is intricately controlled by proteins which constitute the SIN (Septum Initiation Network) signalling cascade. The SIN ensures the coordination between mitotic exit and cytokinesis. Yeast spg1p is a core component of the SIN pathway and we have previously characterized the two orthologs of this G-protein in Arabidopsis thaliana (named AtSGP1 and 2). In this work, the cell and tissue expression of AtSGP genes during plant development has been analysed using AtSGP promoter::GUS fusions in stably transformed A. thaliana lines. AtSGP1 promoter activity was restricted to the quiescent centre, collumella cells, stomata guard cells and the stele while AtSGP2 promoter activity was detected in atrichoblasts, trichomes and pollen. The observed promoter activities are in accordance with publicly available pollen, stomata guard cell and root transcriptome data. Two-hybrid experiments previously evidenced an interaction between AtMAP3Kepsilon1 and AtSGP1. The AtMAP3Kepsilon1 promoter activity was detected in root apices, trichomes and ovule integuments. A genetic approach involving both markers of these specialized cells and mutant backgrounds was used to reinforce our hypothesis. It appears that, although highly conserved between plants and fungi, the spg1p G-protein has evolved in plants to perform a function different from the SIN pathway. Interestingly, cells expressing AtSGPs possessed limited or null mitotic activity. Our data suggests that AtSGP are crucial signalling components involved either in early cell fate specification, or in the final steps of cell differentiation. This is an interesting starting point for a wider study devoted to functional experiments designed to test these hypotheses. PMID- 19123142 TI - Clinical and microbiological efficacy of levofloxacin administered three times a day for the treatment of bacterial conjunctivitis. AB - PURPOSE: This randomized, investigator-masked study aimed to compare the clinical and microbiological effectiveness of three times daily administration of levofloxacin 0.5% eyedrops with the classic, more frequent dosing in patients with bacterial conjunctivitis. METHODS: A total of 120 patients with symptoms of bacterial conjunctivitis were enrolled. The patients were randomly assigned to receive 0.5% levofloxacin eyedrops three times daily to each eye for 5 days (experimental dosage group), or every 2 hours on days 1 and 2, and then every 4 hours on days 3-5 (up to four times per day) (classic dosage group). Ocular symptoms and signs were assessed on day 1, days 3 to 4, and 7 +/- 1 visits. Conjunctival cultures were obtained at baseline and final visits. Clinical outcomes were based on resolution of cardinal signs. Microbial outcomes were based on culture results. RESULTS: Eighty-six patients (41 experimental dosage, 45 classic dosage) were evaluated. There was no difference between the groups in frequency of patients with clinical outcome resolved (85.4% in experimental vs 93.3% in classic dosage group, p=0.3). The microbial eradication rates did not differ statistically between the groups (92.7% vs 95.6%, respectively, p=0.67). CONCLUSIONS: There was no statistically significant difference in the efficacy or safety between the two methods of drug administration. Analysis of the results of compliance supported our conclusion that the less frequent method of dosing of 0.5% levofloxacin eyedrops was more convenient for patients and resulted in better adherence to the drug-dosing scheme. PMID- 19123143 TI - Mild topographic abnormalities that become more suspicious on Scheimpflug imaging. AB - PURPOSE: Although several screening methods exist, postoperative corneal ectasia after refractive surgery is a severe complication. One possibility for this might be the fact that screening methods may fail in detection of preoperative risk factors such as forme fruste keratoconus (FFKC). METHODS: Retrospective evaluation of four cases that showed only mild changes of FFKC on placido-based topography but revealed indicative findings on Scheimpflug imaging (Pentacam). RESULTS: While in placido-based topography evaluation of corneal topography did not show a clear FFKC, the evaluation of corneal topography on Scheimpflug imaging together with the data of spatial corneal thickness revealed distinctive FFKC in all cases presented. CONCLUSIONS: Although both methods bear the risk of not detecting pre-existing FFKC, Scheimpflug imaging seems superior to placido based corneal topography alone. PMID- 19123144 TI - Phakic iris-claw IOL implantation, with Bursa technique, through a self-sealing scleral tunnel and without iridotomy or iridectomy. AB - PURPOSE: To present a novel technique for the implantation of phakic iris-claw intraocular lenses (IOL) in highly myopic cases. METHODS: Twelve cases, under the age of 40, with high myopia were included in this retrospective study. The mean follow-up span was 14.4+/-5.8 months. Phakic iris-claw IOL implantation was performed, through a 5.5 or 6.5 mm self sealing scleral tunnel with Bursa technique. No suturing and opening a peripheral iridectomy or iridotomy was required in our study. Pre- and postoperative patient evaluation included manifest and cycloplegic refractions, uncorrected visual acuity, best-corrected visual acuity (BCVA), induced astigmatism, and intraocular pressure (IOP) assessments. RESULTS: All eyes gained one to nine lines of BCVA of the Snellen chart. The vector analysis revealed an induced astigmatism of 0.63 D. No significant IOP change was detected throughout the follow-up. CONCLUSIONS: The data suggest that nonfoldable iris fixated phakic IOL implantation, through a self-sealing incision with Bursa technique, is safe and requires no iridectomy or iridotomy in cases with deep anterior chambers. PMID- 19123145 TI - Simultaneous surgery in bilateral congenital cataract. AB - PURPOSE: To assess the efficiency of simultaneous surgery in bilateral congenital cataract not only in patients with a high anesthesiologic risk profile. METHODS: A retrospective study was carried out on 40 bilateral congenital cataract patients (80 eyes) who presented to the outpatient department of pediatric ophthalmology of the University Federico II of Naples in the period from 1990 to 2005. All patients had undergone cataract extraction from both eyes in a single surgical session. Visual rehabilitation was achieved in all patients by corneal lenses; successively 25 patients had a secondary intraocular lens implant in the posterior chamber after a period of 2.5-3 years. RESULTS: The mean age at cataract surgery was 7 months (1-17 months). Visual acuity (VA) was assessed in 52 eyes: 8 eyes (15%) presented VA or=20/40. Ocular motility disorders were seen in 28 patients (70%). Eight patients had postoperative complications: the formation of secondary membranes in five patients, and secondary glaucoma in three patients. There were no cases of endophthalmitis. CONCLUSIONS: Simultaneous surgery in bilateral congenital cataract may be taken into consideration not only for patients with a high anesthesiologic risk profile. PMID- 19123146 TI - Scanning electron microscopy and energy dispersive spectroscopy findings in explanted PMMA and hydrophilic acrylic intraocular lenses. AB - PURPOSE: To investigate the presence of calcium (Ca) aggregates influencing biocompatibility and the factors that affect calcium accumulation in explanted intraocular lenses (IOL) and to analyze the Ca distribution in an opacified hydrophilic acrylic lens. METHODS: Surface irregularities and aggregates of 13 IOLs were studied with scanning electron microscopy, and their relative concentrations with energy dispersive spectroscopy (EDS). Relationships of distribution between Ca and silicone (Si) and nitrogen (N) and between N and Si and Na, and the influence of Si on Ca accumulation and the effect of differences in lens material on the distribution of N, and the effect of endophthalmitis on the distribution of Ca were evaluated statistically. EDS analyses were performed on the surface and cross-section of the opacified lens. RESULTS: The statistically significant relationships between the distribution of Ca and N, and between the distribution of N and Na, the significant effect of Si on the Ca accumulation, significant relationship between endophthalmitis and the Ca accumulation in the aggregates were shown. The EDS analysis of the opacified IOL, Ca and P peaks were shown from the whole surface, Ca, O peaks were determined from cross-sections over a 70-80 mum distance. CONCLUSIONS: In the aggregates influencing IOL biocompatibility, presence of proteins was determined to be more important than the presence of Si regarding the distribution of Ca, while the presence of Si affected the accumulation of Ca. Opacification, caused by the Ca accumulation within the lens, was found to result from Ca penetrating from lens pores. PMID- 19123147 TI - Clinical complications of combined phacoemulsification and vitrectomy for eyes with coexisting cataract and vitreoretinal diseases. AB - PURPOSE: To discuss the intraoperative and postoperative complications of combining phacoemulsification and foldable intraocular lens (IOL) implantation with pars plana vitrectomy in eyes with significant cataract and coexisting vitreoretinal diseases. METHODS: This retrospective study consisted of 186 eyes of 149 patients with various vitreoretinal abnormalities and visually significant cataracts. Vitreoretinal surgery was combined with clear corneal phacoemulsification and foldable IOL implantation. Main outcome measures were the intraoperative and postoperative complications at from 6 to 56 months. RESULTS: The most common intraoperative complication was iatrogenic retinal hole (5.3%), transient corneal edema (3.2%), and posterior capsule break (2.1%). The most common postoperative complication was posterior capsule opacification (21.5%) and elevated intraocular pressure (9.7%), macular edema (8.1%), fibrinous reaction (6.9%), vitreous hemorrhage (3.7%), posterior synechiae (3.7%), and recurrent retinal detachment (3.2%). Postoperatively, in 162 eyes (87.1%), visual acuity improved by 3 lines or more on the Snellen chart. In 14 eyes (7.5%), vision remained within 3 lines of preoperative levels and in 10 eyes (5.3%), vision had decreased at the last follow-up. CONCLUSIONS: Postoperative complications did not increase significantly in the combined phacoemulsification and vitreoretinal surgery. Combined vitreoretinal surgery and phacoemulsification with foldable IOL implantation is safe and effective in treating vitreoretinal abnormalities coexisting with cataract. Based on extensive experience with the combined procedure, the authors suggest that combined surgery is recommended in select patients having simultaneous vitreoretinal pathologic changes and cataract. PMID- 19123148 TI - Two-site phacotrabeculectomy versus bimanual microincision cataract surgery combined with trabeculectomy. AB - PURPOSE: To compare the results of two-site phacotrabeculectomy with microincision cataract surgery (MICS) and trabeculectomy. METHODS: Fifty-eight eyes with coexisting cataract and glaucoma were randomly assigned to have a trabeculectomy in a superior quadrant combined with phacoemulsification and intraocular lens (IOL) implantation through temporal clear corneal incision (two site phacotrabeculectomy group), or a trabeculectomy in a superior quadrant combined with MICS followed by IOL implantation from the trabeculectomy incision (MICS + trabeculectomy group). The main outcome measures were preoperative and postoperative intraocular pressure (IOP), number of glaucoma medications, best corrected visual acuity (BCVA), and complications. Follow-up was period was 12 months. RESULTS: Twenty-eight two-site phacotrabeculectomies and 30 MICS + trabeculectomies were performed. Mean age of the subjects was 67.4 years. Mean preoperative IOP was 24.6+/-5.7 mmHg in the two-site phacotrabeculectomy group and 23.7+/-5.5 mmHg in the MICS + trabeculectomy group (p=0.52). At 12 months postoperatively mean IOP was 14.5+/-3.0 and 14.3+/-3.4 mmHg in two-site phacotrabeculectomy and MICS + trabeculectomy groups, respectively (p=0.8). Preoperative and postoperative glaucoma medication numbers were similar and mean number of glaucoma medications decreased in both groups after the surgery (p<0.05). BCVA improved in both groups. Twenty-six of 28 eyes (92%) in the two site + trabeculectomy group and 27 of 30 (90%) in the MICS + trabeculectomy group had clinically apparent filtering blebs at 12 months (p=0.71). Postoperative complications were comparable. CONCLUSIONS: MICS + trabeculectomy provided 1 year IOP control comparable to that with two-site phacotrabeculectomy with similar amount of complications and similar final BCVA. PMID- 19123149 TI - Assessment of cup-to-disc ratio with slit-lamp funduscopy, Heidelberg Retina Tomography II, and stereoscopic photos. AB - PURPOSE: To compare optic disc measurements achieved by slit-lamp funduscopy, Heidelberg Retina Tomography II (HRT II), and stereoscopic optic nerve photos (SONP) in glaucomatous, ocular hypertensive, and normal eyes. METHODS: A total of 176 eyes (glaucomatous [n=87], ocular hypertensive [n=40], and normal eyes [n=49]) of 90 participants were studied. Each participant underwent a full ophthalmic examination, including automated perimetry, slit-lamp funduscopy, HRT II, and SONP. To compare the three different methods being investigated, the following measurements were used: vertical cup to disc ratio (VCDR), horizontal cup to disc ratio (HCDR), and cup to disc area ratio (CDR). Slit-lamp funduscopy was evaluated only with respect to VCDR. HRT II and SONP were evaluated with all three measurements (VCDR, HCDR, and CDR). RESULTS: CDR measurements in ocular hypertensive eyes did not differ significantly according to the measurement methods used (p=0.4). CDR in the other groups of participants, as well as VCDR and HCDR, all differed significantly within each group according to the method used (p<0.05 for all). Mean VCDR measured with funduscopy was smaller than mean VCDR measured with either HRT II or SONP in the glaucoma and ocular hypertension groups (p=0.0001). However, overall, the three methods showed a strong correlation in terms of VCDR, HCDR, and CDR measurements. CONCLUSIONS: Despite the correlation among the three methods, the significant differences between their measurements of optic disc parameters may be too large for these methods to be used interchangeably in clinical situations. PMID- 19123150 TI - Factors affecting intraocular pressure measured by noncontact tonometer. AB - PURPOSE: To investigate the mean values of intraocular pressure (IOP) measured with noncontact tonometer (NCT) and evaluate the factors that may affect IOP. METHODS: A total of 850 subjects who were admitted to our clinic between March 2005 and February 2006 were recruited for the study. Subjects having blepharitis, conjunctivitis, corneal diseases, glaucoma suspicion, or glaucoma were not accepted to the study. All subjects were questioned about systemic diseases. IOP measurement with NCT and central corneal thickness (CCT) with ultrasound pachymetry were performed for each patient between 9 and 11 AM. RESULTS: The mean ages of 367 (43.2%) male subjects and 483 (56.8%) female subjects were 43.9+/ 18.1 and 40.7+/-18.0 years +/- SD, respectively. Since right and left eye IOP, CCT, and keratometric values were significantly correlated, right eye values were used for statistical purposes. Mean IOP values in males and females were 13.2+/ 3.0 and 13.5+/-2.9 mmHg, respectively. Mean CCT values were 552.5+/-34.7 mum for males and 550.1+/-34.3 mum for females. In multiple regression analysis, IOP was found to be associated with gender, refractive error, CCT, and the presence of diabetes mellitus (DM). CONCLUSIONS: Gender, CCT, the presence of DM, and refractive error may be significantly associated with IOP in this particular population. PMID- 19123151 TI - Efficacy of the fixed combinations of bimatoprost or latanoprost plus timolol in patients uncontrolled with prostaglandin monotherapy: a multicenter, randomized, investigator-masked, clinical study. AB - PURPOSE: To compare the efficacy and tolerability of a once-daily evening dose of bimatoprost/timolol fixed combination (BTFC) with that of a once-daily evening dose of latanoprost/timolol fixed combination (LTFC) in patients not controlled with prostaglandins analogues monotherapy. METHODS: A total of 82 patients on prostaglandin analogues monotherapy were enrolled in this prospective, multicenter, investigator masked, clinical study and were randomized to either BTFC (n=47) or LTFC (n=35) topical therapy once at night for 12 weeks. The primary endpoint of the study was to compare the mean daily intraocular pressure (IOP) reduction from baseline between the two treatment arms. Secondary endpoints included the mean daily IOP at 1 and 3 months compared to baseline and the percentage of patients showing a mean IOP reduction from baseline greater than or equal to 15% or 20%. RESULTS: Mean IOP at baseline was 22.7+/-2.0 and 22.1+/-2.6 mmHg in the BTFC and LTFC groups, respectively (p=0.23). Both treatments were effective in reducing the IOP from baseline. The mean IOP reduction was significantly greater in the BTFC group than in the LTFC group (-21.4% vs -13.7%, p<0.001). A higher percentage of patients in the BTFC group showed a mean IOP reduction from baseline >or=15% (72.3% vs 40.0%) and >or=20% (61.7% vs 17.1%) compared to patients in the LTFC group. CONCLUSIONS: Both BTFC and LTFC were more effective versus the monotherapy with prostaglandin analogues. BTFC demonstrated higher performance than LTFC in terms of relative IOP reduction. PMID- 19123152 TI - Early results of modified nonpenetrating deep sclerectomy and phacoemulsification in the treatment of open angle glaucoma and cataract. AB - PURPOSE: To study the outcome and control of intraocular pressure (IOP) of a combined modified nonpenetrating deep sclerectomy (NPDS) without use of implants and phacoemulsification in patients with primary open angle glaucoma (POAG) and cataract. METHODS: Prospective series of 14 eyes in 14 patients with medically uncontrolled POAG and cataract with significant visual impairment undergoing a modified NPDS and phacoemulsification. The surgery was designed to utilize all four mechanisms of outflow track of aqueous of NPDS without the use of scleral implant and a standardized postoperative management and early Nd:YAG laser goniopuncture at 4 weeks. All patients underwent clinical assessment before and after surgery at day 1, day 7, weeks 2, 3, and 4, and then at 2, 3, and 6 months postoperation. Surgical outcome was assessed in terms of IOP, visual acuity, and the incidence of complications. RESULTS: IOP decreased significantly from a preoperative value of 21.71+/-3.81 mmHg (mean +/- SD) to a postoperative value of 13.14+/-3.73 mmHg (mean +/- SD) (p<0.05, paired t test) at 6 months. The number of antiglaucoma eyedrops needed for control of IOP decreased from 3.28+/-0.91 to zero after operation at 6 months follow-up. The change in IOP pre and post laser goniopuncture was noted with no complication or fluctuation in anterior chamber depth. Visual acuity ranged from finger counting to 6/18 with Snellen chart at 6 meters preoperatively to 6/36 to 6/9 postoperatively. There were no intraoperative complications. There was one recessed conjunctival wound at 1 week. No other postoperative complication was noted. CONCLUSIONS: Modified NPDS without scleral implant with early Nd:YAG laser goniopuncture and phacoemulsification for treatment of patients with POAG and cataract is a safe and effective procedure with complete success at 6 months. PMID- 19123153 TI - Oxidative DNA damage in retinopathy of prematurity. AB - PURPOSE: This study examines the levels of oxidative damage in patients with retinopathy of prematurity (ROP). METHODS: Fifty patients were recruited with a birthweight below 1500 g or gestational age below 32 weeks. The cases were classified into those who developed ROP (n=25) and those without ROP (n=25). The authors obtained blood and urine samples from each infant, for measuring 8 hydroxy 2-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels, at the time of the first examination at 4-6 postnatal weeks. RESULTS: A significant difference was observed in leukocyte and urine 8-OHdG levels in patients with ROP compared to those without ROP (p<0.001 for both). Similarly, a significant difference was observed in plasma and urine MDA levels in patients with ROP compared to those without ROP (p<0.001 for both). In addition, significant correlations were found between levels of 8-OHdG in leukocyte DNA and plasma MDA (r=0.859, p<0.001), and between levels of urine 8-OHdG excretion and urine MDA (r=0.563, p<0.001). CONCLUSIONS: 8-OHdG in leukocyte DNA and urine levels in premature infants can be useful as an indicator for ROP screening. PMID- 19123154 TI - Hypoxia tolerance and retinal vein occlusion: a pilot evaluation. AB - PURPOSE: To determine if hypoxia tolerance in patients with retinal vein occlusion (RVO) following exposure to transient hypoxia is different from the hypoxia tolerance of healthy patients without retinal vein occlusion. METHODS: Consecutive patients presenting with RVO following exposure to transient hypoxia (Group I) were compared with healthy subjects (Group II). In addition to cardiovascular and plasma tests, functional respiratory evaluation was performed at rest and during exercise at both normal oxygen levels (21% O2) and in hypoxia (11.6% O2). We used the Wilcoxon test for statistical analysis. RESULTS: Both groups of eight males had similar mean ages: Group I, 47.5 years and Group II, 53 years. In Group I, three patients had glucose or lipid abnormalities, one had hypertension, and one minor thalassanemia. In Group II, one patient had hypertension. At rest in hypoxia, the oxyhemoglobinic desaturation was significantly different (p=0.03) in Group I in comparison with Group II (-13.8 versus -9.3). At exercise in hypoxia, the oxyhemoglobinic desaturation was similar in both groups but there was a statistically significant increase in both systolic (189 versus 155 mmHg; p=0.01) and diastolic (94 versus 77 mmHg; p=0.03) blood pressure in Group I. Ventilation rate and increased heart rate during hypoxia were higher in Group I compared with Group II but were not statistically significant. CONCLUSIONS: In our pilot study, patients with RVO following exposure to transient hypoxia demonstrated intolerance to hypoxia and were significantly different from healthy subjects in their response to hypoxia. A larger study is required to confirm these preliminary results. PMID- 19123155 TI - Prevalence of diabetic retinopathy among subjects with known diabetes in China: the Beijing Eye Study. AB - PURPOSE: To evaluate the prevalence of diabetic retinopathy and risk factors among patients with self-reported diabetes mellitus in China. METHODS: The Beijing Eye Study, a population-based study on inhabitants aged 40+ years, included 4439 subjects. Fundus photographs of the worse eye from participants with self-reported diabetes were graded. RESULTS: Fundus photographs ready for evaluation and a filled out questionnaire were available for 4127 (93.0%) subjects. The prevalence of self-reported diabetes was 235/4127 (5.7%). Among the subjects with a self-reported diagnosis of diabetes, diabetic retinopathy was detected on the fundus photographs of 86 (37.1%) subjects, with macular edema in 12 (5.2%) subjects, clinically significant macular edema in 6 (2.6%) subjects, and a vision- threatening stage of the retinopathy in 12 (5.2%) subjects. Diabetic retinopathy was associated with rural region (p=0.004), longer duration of diabetes (p=0.009), use of diabetic medications (p=0.02), and lower education background (p=0.003). CONCLUSIONS: Prevalence of diabetic retinopathy among Chinese patients aged 40+ years with a self-reported diagnosis of diabetes is about 37%, with a vision-threatening stage of the retinopathy detected in 5% of the subjects. About 5.7% of the adult Chinese population report on a known diagnosis of diabetes mellitus, with about 15% of these subjects knowing about the presence of diabetic retinopathy. The frequency of known diabetes mellitus is lower in rural regions than in urban regions, while diabetic retinopathy overall and macular edema among the subjects with known diabetes mellitus were significantly more common in the rural group. PMID- 19123156 TI - Nutritional AMD treatment phase I (NAT-1): feasibility of oral DHA supplementation in age-related macular degeneration. AB - PURPOSE: To create a pilot study in order to evaluate the feasibility of a prospective case-control study of oral supplementation with fish oil (docosahexaenoic acid [DHA]; eicosapentaenoic acid [EPA]) in a population with age-related macular degeneration (AMD). METHODS: A homogeneous group of 38 patients with drusenoid pigment epithelial detachment in one eye (PED) without choroidal new vessels (CNV) was selected. A complete ophthalmologic examination, and a complete profile of fatty acids in serum (S) and in red blood cell membranes (RBCM), were recorded at day 0 and month 6. In group 1, 22 patients were orally supplemented with EPA (720 mg/day) and DHA (480 mg/day) during 6 months. In group 2, 16 patients were followed as controls. Nutritional recommendations on fish consumption were given to both groups. RESULTS: In group 1, after 6 months supplementation we observed a significant blood enrichment in EPA (EPA-S: 2.20 vs 0.79, p<0.0001 and EPA-RBCM: 2.24 vs 0.85, p<0.0001) and in DHA (DHA-S: 2.47 vs 1.56, p<0.0001 and DHA-RBCM: 6.47 vs 4.67, p<0.0001). No change was observed in group 2 despite nutritional recommendations. In this short followup, no evolution to CNV was noted in either of the two groups. Neither side effects nor dropouts were observed in either of the groups. DISCUSSION: This study supports the feasibility of a long-term double-masked prospective case control study in an AMD population in order to evaluate a potential benefit from oral supplementation with DHA. PMID- 19123157 TI - Prognostic factors after primary vitrectomy and perfluorocarbon liquids for bullous rhegmatogenous retinal detachment. AB - PURPOSE: To identify prognostic factors for visual acuity and anatomic outcomes associated with bullous rhegmatogenous retinal detachment (RRD) management using primary pars plana vitrectomy, intraoperative perfluorocarbon liquids (PFCLs), and internal gas tamponade. METHODS: The authors studied a consecutive series of 115 eyes (115 patients) with a bullous RRD not complicated by proliferative vitreoretinopathy (PVR) associated with large, multiple, and/or posterior breaks in 58 (50.4%) eyes. All eyes underwent vitrectomy, injection of PFCL, and gas tamponade as the primary procedure. Encircling scleral bands were placed in all cases. The follow-up period ranged from 3 to 60 months (mean 16.6+/-14.1 months). RESULTS: Retinal reattachment was achieved in 92.2% of eyes (106/115) with one operation and in all eyes after a second procedure. PVR was observed in 1 (0.87%) eye and preretinal membranes in 3 (2.6%) eyes. Progression of pre-existing cataract and development of new cataract occurred in 45 (58.4%) of the 77 phakic eyes. The presence of inferior retinal breaks was significantly associated with redetachment after the first procedure (p=0.0156). On univariate analysis, better preoperative visual acuity (p<0.001), macular sparing retinal detachment (p<0.001), and fewer quadrants involved by the detachment (p=0.0015) were significant positive prognostic factors for final visual acuity. Logistic regression analysis highlighted that macular sparing retinal detachment and absence of trauma were associated with better final visual acuity. CONCLUSIONS: Redetachment was associated with the presence of inferior retinal breaks. Visual recovery was dependent on preoperative visual acuity, macular involvement, extent of retinal detachment, and trauma. PMID- 19123158 TI - Induction of durable remission in ocular inflammatory diseases. AB - PURPOSE: To describe a paradigm of care for patients with ocular inflammatory diseases aimed at induction of durable remission. METHODS: Retrospective cohort study. The records of 399 patients with ocular inflammatory diseases treated with systemic immunomodulatory therapy (IMT) at the Massachusetts Eye Research and Surgery Institution were reviewed. Durable remission was defined as control of inflammation in the absence of systemic IMT for at least 1 year. Fifty patients met the inclusion criteria. RESULTS: Mean age was 46+/-22.5 years (range 18-88). All the patients had corticosteroid therapy and failed this therapy before having IMT. Fifty-two percent of the patients had used methotrexate alone or in combination with other medications. Thirty percent of the patients required at least 2 years of therapy with systemic IMT to obtain durable remission, while 44% required 2 to 5 years of therapy to achieve the same. Twenty percent continued to stay in remission, off immunomodulatory drugs, between 2 and 5 years and 18% were in remission for more than 5 years after therapy discontinuation. CONCLUSIONS: IMT can be sight saving in patients. It can be tapered and discontinued successfully without the return of ocular inflammation. Durable drug-free remission is an achievable goal, and should be pursued by ocular inflammatory disease specialists. PMID- 19123159 TI - Identification of a novel mutation in a Chinese family with X-linked ocular albinism. AB - PURPOSE: The purpose of the study was to evaluate the GPR143 gene (G-protein coupled receptor 143) in a Chinese three-generation family with OA1, including four carriers and a proband with clinical features of X-linked ocular albinism. METHODS: The proband underwent a detailed ophthalmologic evaluation. Blood samples of family members were obtained and genomic DNA isolated. Mutational analysis by SSCP and direct sequencing of the GPR143 gene was used to screen all nine exons including the intron/exon junctions. The novel mutation c.943G>T (p.G315X) found in the study was confirmed by DHPLC to exclude the possibility of polymorphism. RESULTS: Ophthalmic features of the proband were characteristic of X-linked ocular albinism. The authors identified a novel nonsense mutation p.G315X on exon 8 that was not found in 100 non-albinism subjects by DHPLC. This novel mutation in the GPR143 gene is predicted to subject to nonsense mediated decay. CONCLUSIONS: The novel mutation p.G315X in the OA1 gene was identified in a Chinese family with ocular albinism, which is predicted to generate a premature stop codon. These findings extend the mutational spectrum of GPR143 gene and will be useful for gene diagnosis and genetic counseling in Chinese OA1 patients. PMID- 19123160 TI - Abuse of vasoconstrictive eyedrops mimicking an ocular pemphigoid. AB - PURPOSE: To describe conjunctival histopathologic alterations induced by excessive chronic astringent use. METHODS: Report of a case with clinical picture, epicutane test results, histologic workup of conjunctival biopsy using conventional staining, and immunohistochemical markers. RESULTS: A 45-year-old man using a phenylephrine preparation hourly for years presented with grotesque eye redness, fornix shortening, and scarring of puncta lacrimalia. Direct and indirect immunofluorescence were negative for ocular pemphigoid. Histology revealed signs of chronic inflammation and neovascularization in the conjunctiva. Symptoms resolved after cessation of therapy. CONCLUSIONS: Chronic abuse of decongestant eyedrops can produce a clinical picture resembling an ocular pemphigoid. Histology suggests that late onset immunoreaction and chronic vasoconstriction cause chronic inflammation and neovascularization, respectively. PMID- 19123161 TI - Orthokeratology associated microbial keratitis. AB - PURPOSE: To report the clinical course, microbiologic findings, treatment, and outcomes of overnight orthokeratology associated microbial keratitis. METHODS: Four cases of overnight orthokeratology associated microbial keratitis are reported. RESULTS: Four patients aged 14-23 years (mean 18 years) who had central or paracentral corneal ulcers were included. Visual acuity at presentation ranged from 20/30 to hand motion. In all cases Pseudomonas aeruginosa was cultured from corneal scrapings or storage solution. In all patients the infection resolved with intensive topical antimicrobial treatment. Final best-corrected visual acuity ranged from 20/25 to 20/200 according to the location, size, and density of the corneal scar, which complicated all cases. CONCLUSIONS: Infectious keratitis is a significant, visual threatening complication of overnight orthokeratology. Eye care practitioners should be aware of this complication and educate their patients of the importance of lens hygiene and prompt medical care when symptoms or signs of keratitis appear. PMID- 19123162 TI - Is microbiological analysis of donor cornea transport culture media necessary? AB - PURPOSE: To investigate microbial contamination of the transport medium. Examination of corneoscleral rims is not included in this series. METHODS: Transport media of 63 consecutive grafts done at Tennent Institute of Ophthalmology, Glasgow, were collected for microbial examination. RESULTS: None of the culture plates showed any growth after prolonged culture, and microscopy was negative in 100% of cases. CONCLUSIONS: Routine culture of transport media may not be necessary. PMID- 19123163 TI - Cataract development in a young patient with lathosterolosis: a clinicopathologic case report. AB - PURPOSE: To report a unique case of cataract in a young patient with lathosterolosis, a singular defect of cholesterol biosynthesis, and to report the clinical results and histopathologic findings after cataract surgery. METHODS: A 7-year-old patient with lathosterolosis, a rare defect of cholesterol biosynthesis, presented with a complex phenotype, including severe mental retardation, liver disease, multiple congenital anomalies, and bilateral posterior subcapsular cataracts. After a severe metabolic decompensation, a worsening in the lens opacity of the left eye occurred. The authors thus performed cataract surgery and made a histopathologic analysis of aspirated lenticular samples. The following examinations were performed at 1 day, 1 week, 3 months, 6 months, 12 months, and 24 months: refraction, biomicroscopy, and fundus evaluation. Visual acuity was not assessable due to lack of patient collaboration. The postoperative follow-up period was 24 months. RESULTS: Histopathologic findings on lenticular fragments revealed the presence of fibers disposed in a honeycomb, samples with homogeneous eosinophilic lens fibers, and other fragments characterized by bulgy elements referable to cortical fibers with degenerative characteristics. After surgery, biomicroscopic evaluation revealed no significant inflammation and good intraocular lens centration at the various control visits. No intraoperative or postoperative complications occurred. No posterior capsule opacification occurred 2 years after surgery. CONCLUSIONS: Lathosterolosis may lead to dysmetabolic cataract development; this unique case of cataract in such a patient has been successfully managed with surgery. Clinical results were excellent, and no complications occurred either intra- or postoperatively. PMID- 19123164 TI - Capsule fixation device for cataract surgery. AB - PURPOSE: The authors present a patient with Marfan syndrome who underwent cataract surgery using a novel device for fixation of capsular bag. METHODS: The capsule fixation device (CFD) is poly(methyl methacrylate) arch with double-armed bent hook that can be introduced into the anterior chamber through the clear corneal incision, placed with a rest on the capsule equator, and sutured to the sclera without injury of the capsular bag. A 16-year-old patient with Marfan syndrome underwent consecutive phacoemulsification with in-the-bag intraocular lens and CFD implantation on both eyes. RESULTS: During the surgery handling the CFD was technically simple with good visualization. The preoperative best corrected visual acuity was 0.3 and 0.1 and postoperatively 0.6 and 0.8 in the right and left eyes, respectively. The postoperative follow-up was over 9 months. CONCLUSIONS: Use of the CFD was successful in a case of lens subluxation with no complications observed during the period of follow-up. PMID- 19123165 TI - Nonpenetrating glaucoma surgery with goniosynechiolysis ab interno: a surgical technique. AB - PURPOSE: To present a surgical technique for treatment of peripheral anterior synechiae (PAS or goniosynechiae) at the operative field during nonpenetrating glaucoma surgery. METHODS: After usual preparation of a superficial and a deep scleral flap with externalization of the Schlemm's canal and peeling the juxtacanalicular trabecular meshwork, the goniosynechia is transected by a spatula introduced to the anterior chamber through a paracentesis. A Descemet's window provides full visual control at the peripheral cornea at the basis of the superficial scleral flap. RESULTS: With this surgical technique, selective treatment of goniosynechiae is possible exactly at the area where creation of low outflow resistance is intended by the surgeon during nonpenetrating glaucoma surgery. CONCLUSIONS: This technique enables the surgeon to perform nonpenetrating glaucoma surgery even in the presence of peripheral anterior synechiae. PMID- 19123166 TI - Choroidal neovascularization in angioid streaks and pseudoxanthoma elasticum: 1 year follow-up. AB - PURPOSE: To report the efficacy of intravitreal injection of bevacizumab 1.25 mg (IVB) in the treatment of choroidal neovascularization (CNV) secondary to angioid streaks (AS). METHODS: Case review of two patients with CNV secondary to AS treated with three IVB with a 1-year follow up. RESULTS: In both patients after 1 year fluorescein angiography and optical coherence tomography showed complete inactivity of the CNV and the ophthalmologic examination revealed stabilization of the visual acuity. CONCLUSIONS: The IBV appeared to be an effective and safe treatment for CNV secondary to AS resulting in a long-term CNV inactivation. Further long-term studies in this type of lesion are desirable. PMID- 19123167 TI - Low-fluence photodynamic therapy in longstanding chronic central serous chorioretinopathy with foveal and gravitational atrophy. AB - PURPOSE: To describe anatomic and functional outcomes in two patients with long standing severe chronic central serous chorioretinopathy (CSC) with foveal and gravitational atrophy treated with low-fluence photodynamic therapy (PDT). METHODS: Two patients with a history of over 10 years of chronic CSC and development of gravitational tracts, with best-corrected visual acuity 99 %). The internal 5-methylene-3,5-dihydroimidazol-4 one (MIO) cofactor is essential for the PAM-catalysed amination reactions. The regioselectivity of amination reactions was influenced by the nature of the ring substituent. PMID- 19123197 TI - Altering the substrate specificity of RhlI by directed evolution. AB - REDUCING VIRULENCE: RhlI catalyzes the synthesis of N-butanoyl homoserine lactone (BHL), with a minor product N-hexanoyl homoserine lactone (HHL). By using directed evolution and a genetic screen, RhlI has been engineered for enhanced production of both BHL and HHL at a similar level. Quorum sensing regulates biofilm formation and virulence factor production in the human opportunistic pathogen Pseudomonas aeruginosa. We used directed evolution to engineer RhlI, an enzyme in the RhlI-RhlR quorum-sensing system of P. aeruginosa, to alter its substrate specificity and gain insight into the molecular mechanisms of quorum sensing. By using a genetic screen, we identified a mutant with improved production of RhlI's two signaling molecules, N-butanoyl- and N-hexanoyl homoserine lactone (BHL and HHL). In particular, production of BHL has been enhanced by more than two-fold, and the synthesis of HHL has been improved from an undetectable level to a level similar to BHL; this change indicates a significant change in substrate specificity. No significant change in the gene expression level was observed. Sequence alignments suggest that the mutations are most likely to facilitate interactions between the enzyme and the two acylated ACP substrates. This work also demonstrates that the genetic screen/selection should be useful in engineering additional quorum-sensing components. PMID- 19123198 TI - Isolation of phospholipase D mutants having phosphatidylinositol-synthesizing activity with positional specificity on myo-inositol. AB - ENZYME-MEDIATED SYNTHESIS OF PHOSPHATIDYLINOSITOL: Engineered phospholipase D enzymes enable the synthesis of phosphatidylinositol by transphosphatidylation. The 1- or 3-hydroxy group of myo-inositol is selectively reacted. Phospholipase D (PLD) mutants that have phosphatidylinositol (PI)-synthesizing activity with positional selectivity towards 1- or 3-OH groups of myo-inositol have been isolated. A mutant PLD library, in which site-directed saturation mutations were introduced in vitro at positions 187, 191, and 385 of the wild-type PLD of Streptomyces antibioticus, was screened for PI-synthesizing mutants. TLC and HPLC analyses of the PI synthesized by the isolated mutant PLDs revealed that three mutants, namely 187D/191Y/385R (DYR), 187A/191Y/385R (AYR), and 187M/191Y/385R (MYR), selectively generated 1- or 3-PI among the other possible PI positional isomers. Taking into account the consensus sequence of the three mutants, a series of mutants, 187X/191Y/385R (XYR), was constructed and analyzed. Almost all the XYR mutants generated 1(3)-PI selectively, thus suggesting that the Y385R mutation contributed to the selectivity for the 1(3)-PI synthesis. The XYR mutants showed similar phosphatidylcholine-hydrolyzing activity among the mutants, but the PI-synthesizing activities were different depending on the amino acid at position 187. In particular, aromatic amino acids at position 187 greatly reduced the PI-synthesizing activity. The ratios of 1-PI versus 3-PI in the PIs synthesized with the XYR mutants were analyzed by selective hydrolysis with PI specific phospholipase C. It was found that 187H/191Y/385R (HYR) generated 1-PI more than 3-PI (ratio=7:3), whereas 187T/191Y/385R (TYR) generated 1-PI less than 3-PI (ratio=2:8). This confirmed that the amino acid at position 187 determined the selectivity between 1-PI and 3-PI formation. PMID- 19123199 TI - Can CSF biomarkers or pre-treatment progression rate predict response to cholinesterase inhibitor treatment in Alzheimer's disease? AB - OBJECTIVE: The main objective of this study was to investigate possible predictors of response to cholinesterase inhibitor (ChEI) treatment, including pre-treatment progression rates and levels of the cerebrospinal fluid (CSF) biomarkers. A secondary objective was to evaluate whether treatment with ChEI changed progression. METHODS: Out-patient individuals (n = 191) with the clinical diagnosis of Alzheimer's disease received ChEI treatment and were part of the Swedish Alzheimer Treatment Study (SATS), a prospective, longitudinal, non randomised study in a routine clinical setting. Patients were assessed with MMSE, ADAS-cog and a global rating (CIBIC) at baseline, 2 months and every 6 months for a total period of 3 years. The following potential predictors of treatment response were investigated: age, gender, APOE epsilon 4 carrier, education, duration of disease, cognitive level, pre-treatment progression rate (in MMSE) and the levels of the CSF biomarkers A beta 42, T-tau and P-tau. RESULTS: Fast pre-treatment progression rate was a predictor of treatment response even after adjusting for baseline severity, another positive predictor of response. Patients in the fastest quartile of pre-treatment progression rates were significantly more prone to be responders at 2 months (adjusted OR 6.6, p = 0.001) and 6 months (adjusted OR 10.4, p < 0.001) than those in the slowest progressing quartile. Moreover, the linearity of progression was significantly changed by ChEI treatment at 6 months compared to the pre-treatment period. CONCLUSION: The rate of pre-treatment progression was the most consistent positive predictor of ChEI treatment response in the routine clinical setting. The progression rate was significantly changed by ChEI treatment. PMID- 19123201 TI - Reprogramming of the transcriptome in a novel chromosome 3 transfer tumor suppressor ovarian cancer cell line model affected molecular networks that are characteristic of ovarian cancer. AB - Tumor suppression as a consequence of the transfer of chromosome 3p fragments was previously observed in a novel epithelial ovarian cancer (EOC) OV-90 cell line model harboring loss of 3p. Microarray analysis revealed that tumor suppression was associated with a modified transcriptome. To investigate the relevance of the altered transcriptome, the differentially expressed genes identified by Affymetrix analysis in the 3p transfer studies, were integrated with a comparative microarray analysis of normal ovarian surface epithelial (NOSE) cells and malignant ovarian (TOV) cancers. Data from 219 significantly differentially expressed genes exhibited patterns in the direction predicted by the analysis of 3p transfer study. The 30 genes with the highest statistically significant differences (P < 1 x 10(-8)) in expression were found consistently differentially expressed between NOSE and TOV samples. The investigation of these genes in benign serous ovarian tumors and EOC cell lines also exhibited predictable expression patterns. Within the group of differentially expressed genes were SPARC, DAB2, CP, EVI1, ELF3, and EHD2, known to play a role in ovarian cancer, genes implicated in other cancers, such as GREM1 and GLIPR1, as well as genes not previously reported in a cancer context such as AKAP2 and ATAD4. A number of the differentially expressed genes are implicated in the TGF-beta signaling pathway. These findings suggest that the reprogramming of the transcriptome that occurred as a consequence of the chromosome 3 transfer and tumor suppression affected molecular networks that are characteristic of ovarian carcinogenesis thus validating our novel ovarian cancer cell line model. PMID- 19123202 TI - Interaction of NO molecules with Pd clusters: ab initio density-functional study. AB - The adsorption of NO molecules on small Pd(n) (n = 1-6) clusters has been studied using first-principles density-functional theory. Three adsorption sites were considered: vertex (on-top), bridge, and hollow. Adsorption is strong, ranging from 2 to 3 eV. In all cases NO adsorbs in a bent configuration. Calculated shifts in N-O bond vibration frequencies (with anharmonic corrections) agree very well with available experimental data. In contrast to metallic Pd surfaces, adsorption of NO on palladium clusters causes considerable changes in geometry around adsorption site because palladium d-orbitals rehybridize to maximize the overlap with NO orbitals (mainly the antibonding pi*). Thus, the overall energetic effect of NO adsorption is the result of two competing processes: lowering of the total energy through tighter bonding with NO and rising the energy due to cluster deformation. The Pd(n)-NO bond creation is governed by electron transfer from Pd-d orbitals into the NO pi*. As a result, the Pd cluster becomes locally demagnetized (with total magnetic moment of 1 micro(B) located at Pd atoms not connected to NO) and the NO molecule is activated: the N-O bond length is increased and the vibration frequency is redshifted. PMID- 19123200 TI - Models for chromosomal replication-independent non-B DNA structure-induced genetic instability. AB - Regions of genomic DNA containing repetitive nucleotide sequences can adopt a number of different structures in addition to the canonical B-DNA form: many of these non-B DNA structures are causative factors in genetic instability and human disease. Although chromosomal DNA replication through such repetitive sequences has been considered a major cause of non-B form DNA structure-induced genetic instability, it is also observed in non-proliferative tissues. In this review, we discuss putative mechanisms responsible for the mutagenesis induced by non-B DNA structures in the absence of chromosomal DNA replication. PMID- 19123204 TI - Charge transport in stacking metal and metal-free phthalocyanine iodides. Effects of packing, dopants, external electric field, central metals, core modification, and substitutions. AB - The charge-transport properties of the one-dimensional stacking metal phthalocyanine iodides (M(Pc)I, M = Fe, Co, Ni, Cu) and metal-free phthalocyanine iodide (H2(Pc)I) have been theoretically investigated. On the basis of the tight binding approximation and two-state theory, both the site-energy corrected energy splitting in dimer and Fock-matrix-based methods are used to calculate the transfer integral. The intermolecular motions, including interplanar translation, rotation, slip, and tilt, exert remarkable impacts on the transfer integral. The order/disorder of the dopant stack and the long-range electrostatic interactions are also demonstrated to be crucial factors for modulation of charge-transport properties. The transfer integral undergoes slight changes under an applied electric field along the stacking direction in the range of 10(6)-10(7) V cm(-1). The change of central metals in MPc has little effect on the transfer integrals, but significantly affects the reorganization energies. The extension of the pi conjugation in macrocyclic ligand brings about considerable influence on the transfer integrals. Peripheral substitutions by animo, hydroxyl, and methyl lead to deviations from planarity of macromolecular rings, and hence affect the valence bands significantly. PMID- 19123203 TI - Rotamer optimization for protein design through MAP estimation and problem-size reduction. AB - The search for the global minimum energy conformation (GMEC) of protein side chains is an important computational challenge in protein structure prediction and design. Using rotamer models, the problem is formulated as a NP-hard optimization problem. Dead-end elimination (DEE) methods combined with systematic A* search (DEE/A*) has proven useful, but may not be strong enough as we attempt to solve protein design problems where a large number of similar rotamers is eligible and the network of interactions between residues is dense. In this work, we present an exact solution method, named BroMAP (branch-and-bound rotamer optimization using MAP estimation), for such protein design problems. The design goal of BroMAP is to be able to expand smaller search trees than conventional branch-and-bound methods while performing only a moderate amount of computation in each node, thereby reducing the total running time. To achieve that, BroMAP attempts reduction of the problem size within each node through DEE and elimination by lower bounds from approximate maximum-a-posteriori (MAP) estimation. The lower bounds are also exploited in branching and subproblem selection for fast discovery of strong upper bounds. Our computational results show that BroMAP tends to be faster than DEE/A* for large protein design cases. BroMAP also solved cases that were not solved by DEE/A* within the maximum allowed time, and did not incur significant disadvantage for cases where DEE/A* performed well. Therefore, BroMAP is particularly applicable to large protein design problems where DEE/A* struggles and can also substitute for DEE/A* in general GMEC search. PMID- 19123205 TI - Stable five-membered-ring allenes with second-row elements only: not allenes, but zwitterions. AB - A pyrazole derivative, considered as the stable five-membered cyclic allene 1 a by Dyker, Bertrand, et al., is reinterpreted to be a zwitterion 1 b. It is shown that the bonding characteristics typical for an allene are not possible for structure 1 a. Ar = 2,6-dimethylphenyl. PMID- 19123206 TI - High-yielding synthesis of the anti-influenza neuramidase inhibitor (-) oseltamivir by three "one-pot" operations. AB - Taking shortcuts: A remarkably short and high-yielding asymmetric total synthesis of (-)-oseltamivir takes advantage of organocatalysis and single-pot domino operations. The target, known as the drug Tamiflu, is prepared efficiently in a short time, and also its derivatives can be synthesized effectively. PMID- 19123207 TI - Enantioselective intermolecular crossed-conjugate additions between nitroalkenes and alpha,beta-enals through a dual activation strategy. AB - Double the fun: The title reaction was developed by using a Lewis base/iminium activation strategy (see scheme). The reaction proceeded with excellent yields and ee values, and the products were additionally transformed into a single enantiomer of a substituted pyrrolidine with excellent retention of configuration. PMID- 19123208 TI - Electronic structure, chemical bonding, and solid-state NMR spectroscopy of the digallides of Ca, Sr, and Ba. AB - Combined application of (69,71)Ga NMR spectroscopy and quantum mechanical calculations reveals the chemical bonding in the digallides of Ca, Sr, and Ba. An analysis of the electron localization function (ELF) shows honeycomb-like 6(3) nets of the Ga atoms as the most prominent structural features in SrGa(2) and BaGa(2). For CaGa(2) a description of a 3+1-coordinated Ga atom is revealed by the ELF and by an analysis of interatomic distances. The NMR spectroscopic signal shift is mainly due to the Knight shift and is almost equal for the investigated digallides, whereas the anisotropy of the signal shift decreases with the radius of the alkaline-earth metals. Calculated and observed values of the electric field gradient (EFG) are in good agreement for CaGa(2) and BaGa(2) but differ by about 21 % for SrGa(2) indicating structural instability. Better agreement is achieved by considering a puckering of the Ga layers. For BaGa(2) an instability of the structure is indicated by a peak in the density of states at the Fermi level, which is shifted to lower energies when taking puckering of the Ga layers into account. Both structural modifications are confirmed by crystallographic information. The Fermi velocity of the electrons is strongly anisotropic and is largest in the (001) plane of the crystal structure. This results in an alignment of the crystallites with the [001] axis perpendicular to the magnetic field as observed in (69,71)Ga NMR spectroscopy and magnetic susceptibility experiments. The electron transport is predominantly mediated by the Ga-Ga p(x)- and p(y)-like electrons in the (001) plane. The specific heat capacity of BaGa(2) was determined and indicated the absence of phase transitions between 1.8 and 320 K. PMID- 19123209 TI - Regio-, peri-, and torquoselectivity in hydroxy heptatrienyl cation electrocyclizations: the iso/homo-Nazarov reaction. AB - The electrocyclizations of 1- and 3-hydroxyheptatrienyl cations have been computationally studied at the B3LYP/6-311G(d) level. The 1-hydroxy system clearly favors a 4pi e(-) over a 6pi e(-) process, while for the 3-hydroxy isomer these mechanisms compete. Substituents can either enhance or invert this periselectivity through steric or electronic effects, respectively. Houk's model of torquoselectivity helps to explain the activation energy differences between the alternative 4pi e(-) electrocyclizations available for each system. The cyclopentenyl cations thus obtained can evolve, either through intramolecular trapping by the vinyl group or by 1,3-proton migration, along reaction coordinates which could correspond to either very asynchronous concerted mechanisms, or two-step reactions with very shallow or non-existent intermediates. PMID- 19123210 TI - Trading off SNR and resolution in MR images. AB - With a fixed time to acquire a magnetic resonance (MR) image, time can be spent to acquire better spatial resolution with decrease in signal-to-noise ratio (SNR) or decreased resolution with increase in SNR. This resolution/SNR tradeoff at fixed time has been investigated by a visual rater study using images of ex vivo mouse brains. Simulated images with a tradeoff between SNR and resolution were produced from high-quality, 3D isotropic mouse brain images to emulate shorter constant acquisition times. The tradeoff images spanned a range of SNRs (63-6) and isotropic resolutions (32-81 microm). Fourteen readers identified the image which best displayed neuroanatomy. Additional experiments tested for (i) intra observer consistency, (ii) the effect of emulated scan time, and (iii) specifically biased questions pertaining to the perception of neuroanatomy. Optimal anatomical viewing depended primarily on the SNR of the images. Specifically, for fixed imaging time, preference lay in the SNR range of approximately 30-35 with strong consistency and there was minimal effect from overall imaging time. PMID- 19123211 TI - Studies on the efficient generation of phosphorus-carbon bonds via a rearrangement of P(III) esters catalysed by trimethylhalosilanes. AB - Halotrimethylsilanes Me(3)SiX (X = Br, I) catalyse rearrangements of tricoordinate phosphorus esters R'R''P-OR into the corresponding phosphoryl systems R'R''P(O)R. This provides a simple and efficient route to a variety of structures containing phosphorus-carbon bonds, under mild conditions and with good yields. The reaction mechanism was investigated in detail by (31)P NMR spectroscopy and independent synthesis of the reaction intermediates. It has been demonstrated that the primary products of this catalytic reaction are halogeno P(III) structures R'R''PX and silyl ethers ROSiMe(3) and that they subsequently react to give the corresponding phosphorus silyl esters-Me(3)SiOPR'R''-and alkyl halides RX. At higher temperatures these intermediates then react to form R'R''P(O)R compounds. This paper also features the surprising observation that when esters Ph(2)POR and halotrimethylsilanes Me(3)SiX (X = Br, I) are used in 2:1 ratio, phosphonium salts Ph(2)R(2)P(+)X(-) and trimethylsilyl diphenylphosphinate--Ph(2)P(O)OSiMe(3)--are formed as the major products. Experimental evidence indicates that the mechanisms of both reactions are fundamentally different from that of the Michaelis-Arbuzov reaction. Me(3)SiCl is not reactive and this paper explains why. PMID- 19123212 TI - Density functional study of proline-catalyzed intramolecular Baylis-Hillman reactions. AB - The mechanisms of proline-catalyzed and imidazole-co-catalyzed intramolecular Baylis-Hillman reactions have been studied by using density functional theory methods at the B3LYP/6-31G(d,p) level of theory. A polarizable continuum model (PCM B3LYP/6-31++G(d,p)//B3LYP/6-31G(d,p)) was used to describe solvent effects. Different reaction pathways were investigated, which indicated that water is an important catalyst in the imine/enamine conversion step in the absence of imidazole. When imidazole is used as a co-catalyst, water is still important in the imidazole addition step, but is not present in the Baylis-Hillman cyclization step. The computational data has allowed us to rationalize the experimental outcome of the intramolecular Baylis-Hillman reaction, validating some of the mechanistic steps proposed in the literature, as well as to propose new ones that considerably change and improve our understanding of the full reaction path. PMID- 19123213 TI - Tuning single-molecule dynamics in functionalized mesoporous silica. AB - Mesoporous silica materials are promising host structures for diverse applications in nanoscience. Many applications can profit significantly from the ability to influence guest dynamics in the host matrix. To this end, we introduce covalently attached organic functionalization into the walls of mesoporous silica networks. Using single-molecule fluorescence microscopy, we study the diffusion behavior of single terrylene diimide dye molecules in functionalized mesoporous silica films. We show that, through variation of the chemical nature and density of the functional groups, the diffusion dynamics of the dye molecules, in the presence of the surfactant template, can be controlled precisely. The mean diffusion coefficient of the dye molecules increases or decreases depending on the functional group attached to the silica wall. This allows fine-tuning of the diffusion dynamics of the dye by approximately one order of magnitude. The observed changes in the mean diffusion coefficients can be explained by shielding of hydroxyl groups on the silica surface in combination with changes in the rigidity of the micellar packing in the film, as well as direct interactions between the functional groups and the dye molecules. PMID- 19123214 TI - Speciation of rare-earth metal complexes in ionic liquids: a multiple-technique approach. AB - The dissolution process of metal complexes in ionic liquids was investigated by a multiple-technique approach to reveal the solvate species of the metal in solution. The task-specific ionic liquid betainium bis(trifluoromethylsulfonyl)imide ([Hbet][Tf(2)N]) is able to dissolve stoichiometric amounts of the oxides of the rare-earth elements. The crystal structures of the compounds [Eu(2)(bet)(8)(H(2)O)(4)][Tf(2)N](6), [Eu(2)(bet)(8)(H(2)O)(2)][Tf(2)N](6) x 2 H(2)O, and [Y(2)(bet)(6)(H(2)O)(4)][Tf(2)N](6) were found to consist of dimers. These rare earth complexes are well soluble in the ionic liquids [Hbet][Tf(2)N] and [C(4)mim][Tf(2)N] (C(4)mim = 1-butyl-3-methylimidazolium). The speciation of the metal complexes after dissolution in these ionic liquids was investigated by luminescence spectroscopy, (1)H, (13)C, and (89)Y NMR spectroscopy, and by the synchrotron techniques EXAFS (extended X-ray absorption fine structure) and HEXS (high-energy X-ray scattering). The combination of these complementary analytical techniques reveals that the cationic dimers decompose into monomers after dissolution of the complexes in the ionic liquids. Deeper insight into the solution processes of metal compounds is desirable for applications of ionic liquids in the field of electrochemistry, catalysis, and materials chemistry. PMID- 19123215 TI - Solid-phase synthesis of peptide libraries combining alpha-amino acids with inorganic and organic chromophores. AB - The synthesis of two series of peptidic chains composed of bis(terpyridine)ruthenium(II) acceptor units and organic chromophores (coumarin, naphthalene, anthracene, fluorene) by stepwise solid-phase peptide synthesis (SPPS) techniques is described. The first series of dyads comprises directly amide linked chromophores, while the second one possesses a glycine spacer between the two chromophores. All dyads were studied by UV/Vis and NMR spectroscopy, steady-state luminescence, luminescence decay and electrochemistry, as well as by DFT calculations. The results of these studies indicate weak electronic coupling of the chromophores in the ground state. Absorption spectra of all dyads are dominated by metal-to-ligand charge-transfer (MLCT) bands around 500 nm. The bichromophoric systems, especially with coumarin as organic chromophore, display additional strong absorptions in the visible spectral region. All complexes are luminescent at room temperature ((3)MLCT). Efficient quenching of the fluorescence of the organic chromophore by the attached ruthenium complex is observed in all dyads. Excitation spectra indicate energy transfer from the organic dye to the ruthenium chromophore. PMID- 19123217 TI - [M(N3)2(H2O)2] x (bpeado): unusual antiferromagnetic Heisenberg chain (M = Mn) and ferromagnetic Ising chain (M = Co) with large coercivity and magnetic relaxation (bpeado = 1,2-bis(4-pyridyl)ethane-N,N'-dioxide). AB - We present here the structures and magnetism of two isostructural magnetic metal azido chain compounds of the type [M(N(3))(2)(H(2)O)(2)](bpeado) (1, M = Mn; 2, M = Co; bpeado = 1,2-bis(4-pyridyl)ethane-N,N'-dioxide), prepared by utilizing the long spacer bpeado. The structure is composed of [M(N(3))(2)(H(2)O)(2)](n) metal azido chains, in which metal ions are bridged by two end-on azido ligands, and the chains are further supported by H-bonding interactions between the coordination water of the chain and the lattice bpeado in a three-dimensional (3D) threefold interpenetrated H-bonded framework. Within the structure, the metal-azido chains are separated quite well. Investigation of the magnetic properties has revealed that the Mn compound 1 is an isotropic Heisenberg chain with unusual antiferromagnetic coupling between the Mn(2+) ions linked by double end-on azide anions. The Co compound 2 shows a strong anisotropic Ising-type ferromagnetic chain within the material. Detailed magnetic studies on both polycrystalline and single-crystal samples of 2 have revealed a large coercivity of up to 37.5 kOe (along the crystallographic b-axis), multi-magnetic transitions, and single-chain-magnet-like magnetic relaxation behavior. PMID- 19123216 TI - Total syntheses of casuarine and its 6-O-alpha-glucoside: complementary inhibition towards glycoside hydrolases of the GH31 and GH37 families. AB - Total synthesis of naturally occurring casuarine (1) and the first total synthesis of casuarine 6-O-alpha-glucoside (2) were achieved through complete stereoselective nitrone cycloaddition, Tamao-Fleming oxidation and selective alpha-glucosylation as key steps. Biological assays of the two compounds proved their strong and selective inhibitory properties towards glucoamylase NtMGAM and trehalase Tre37A, respectively, which place them among the most powerful inhibitors of these enzymes. The structural determination of the complexes of NtMGAM with 1 and of Tre37A with 2 revealed interesting similarities in the catalytic sites of these two enzymes which belong to different families and clans. PMID- 19123218 TI - A synthetic method for transition-metal chalcogenide nanocrystals. AB - In this paper, we prepared a series of chalcogenide semiconductor nanocrystals in controllable shape and size via a facile wet route using metal nitrates and sulfur or selenium powder as precursors and octadecylamine (ODA) as solvent. The as-obtained chalcogenides included CdS, MnS, Ag(2)S, PbS, Cu(1.8)S, Bi(2)S(3), ZnS, Zn(x)Cd(1-x)S, as well as Ag(2)Se, Cu(2-x)Se, CdSe, MnSe. Furthermore, these cyclohexane-soluble monodisperse nanocrystals were assembled to water-soluble colloidal spheres and the adjustment of assembly orderliness has been achieved by controlling the experimental parameters. The general synthesis and assembly of chalcogenide semiconductors provide ideal building blocks for various potential applications. PMID- 19123219 TI - Efficient air-stable organometallic low-molecular-mass gelators for ionic liquids: synthesis, aggregation and application of pyridine-bridged bis(benzimidazolylidene)-palladium complexes. AB - Novel pincer-type, pyridine-bridged bis(benzimidazolylidene)-palladium complexes 5-7 were synthesised from cheap commercial precursors under microwave assistance. Although simple in structure, carbene complexes 5a,b are efficient low-molecular mass metallogelators. They gelate not only a broad variety of protic and aprotic organic solvents, but also different types of customary ionic liquids (such as imidazolium, pyridinium, pyrazolidinium, piperidinium and ammonium salts) at concentrations as low as 0.5 mg mL(-1). The morphologies of the resulting 3D gel networks composed from long and thin fibres were studied by TEM and light microscopy for a selection of organic and ionic liquids. The achiral gelators are able to induce the formation of helical fibres. The thermal stability of the gel samples increases with the gelator concentration as demonstrated by thermoreversible DSC studies. Temperature-dependent NMR and X-ray diffraction studies, as well as comparisons with pincer complex analogues bearing shorter alkyl chains, suggest that the 3D networks responsible for gelation are based on non-covalent interactions, such as pi-stacking, van der Waals interactions, and hydrogen and metal-metal bonding. Ionic liquids and gels obtained from them and 5a,b display comparable high conductivities, which characterises pyridine-bridged bis(benzimidazolylidene)-palladium pincer complexes as air-stable metallo gelators that efficiently immobilise ionic liquids in low gelator concentration indicating--beyond catalysis--their potential applications in electrochemical devices. PMID- 19123220 TI - Stereoselective allylation of ketones: explanation for the unusual inversion of the induced stereochemistry in the auxiliary-mediated crotylation and pentenylation of butanone by DFT calculations. AB - Auxiliary-mediated domino crotylations and pentenylations of butanone yield homoallylic ethers with two newly formed stereogenic centers. With our norpseudoephedrine-derived auxiliary, we observed the formation of anti isomers exclusively, and the nature of the major isomer was independent of the substrate double bond geometry. Interestingly, there is a switch in induced selectivity when going from crotylation to pentenylation. Here, we present the computational rationalization for this behavior by identification of the relevant transition states (TSs), the energies of which were determined by using the B3LYP/6-31+G(d) level of theory in combination with the PCM/UAKS method to include the effects exerted by the solvent dichloromethane. To quickly narrow down the number of potentially relevant TSs from the whole set of 288 and 864 TSs for the crotylation and pentenylation, respectively, we employed a screening process based on B3LYP//AM1 energies. The predicted selectivities are in good agreement with experimentally determined ones. Furthermore, the obtained results also facilitate an explanation of the selectivities obtained in hexenylations and heptenylations. Finally, activation energies were determined that account for the significantly longer reaction times than those for the domino allylation with unsubstituted trimethylallylsilane. PMID- 19123221 TI - Supramolecular assembly of self-labeled amphicalixarenes. AB - The synthesis and precise supramolecular organization of new amphicalixarene 4 bearing four rodlike aligned fluorescent terephthalic benzamide moieties in the upper rim is reported. The aggregation of 4 was monitored by a combination of fluorescence, conductometry, and cryo-TEM measurements at different pH values and revealed significant structural differences. Most interestingly, we found that exactly 12 molecules of 4 assemble to form a spherical and structurally persistent micelle at pH 7, which coexists with rodlike micelles. In contrast to other examples of structurally defined micelles reported previously, this new type of self-labeled amphiphile serves as a fluorescence reporter with the terephthalic units additionally providing an extended cavity structure in the hydrophilic part, which facilitates the inclusion of guest molecules. In addition to the observed formation of well-defined micelles, the other highly important finding from this study is the fact that guest molecules directly influence the micellar organization because they can interact with both the free amphiphiles below the critical micelle concentration (cmc) or the micellar aggregate itself. These two types of interactions are especially pronounced in the case of the cationic pyrene derivative 10, which binds electrostatically with the amphiphile 4. In addition, a very unique membrane structure exhibiting a regular hexagonal pattern of 5 nm pores is formed by 4 at pH 4. PMID- 19123222 TI - Enantioselective and catalytic method for alpha-crotylation of aldehydes with a kinetic self-refinement of stereochemistry. AB - Kinetic refinery: A practical, highly stereoselective, two-step catalytic protocol for the alpha-allylation of aldehydes, starting from crotyltrichlorosilanes, has been developed (see scheme). In each reaction step, one of the stereoisomers reacted faster than the other, which resulted in a kinetic stereochemical (E/Z) self-refinement of the system and led to the formation of virtually enantiomerically and geometrically pure linear homoallylic alcohols in high yield. PMID- 19123223 TI - Nanoparticle-supported and magnetically recoverable ruthenium hydroxide catalyst: efficient hydration of nitriles to amides in aqueous medium. AB - Magnetic attraction not filtration: A magnetic nanoparticle-supported ruthenium hydroxide catalyst (see figure) was readily prepared from inexpensive starting materials and shown to catalyze the hydration of nitriles with excellent yield in a benign aqueous medium. Catalyst recovery using an external magnetic field, superior activity, and the inherent stability of the catalyst system are additional sustainable attributes of this protocol. PMID- 19123224 TI - Highly enantioselective conjugate additions of aldehydes to vinyl sulfones. AB - Joined together, organocatalysts aldehydes and sulfones: A diaryl prolinol silyl ether was found to catalyse efficiently and enantioselectively the conjugate addition of aldehydes to vinyl sulfones (see scheme). The ample synthetic utility of the resulting adducts is illustrated. PMID- 19123225 TI - Improved palladium-catalyzed Sonogashira coupling reactions of aryl chlorides. PMID- 19123226 TI - N4-tetradentate dicarboxyamidate/dipyridyl palladium complexes as robust catalysts for the Heck reaction of deactivated aryl chlorides. AB - Structurally well defined and thermally stable Pd(II) complexes, derived from N4 tetradentate dicarboxyamide/dipyridyl ligands, were evaluated as catalysts for the Heck reactions of deactivated aryl chlorides and olefins (see scheme). The concept of using an anionic carboxyamide as an ancillary ligand for palladium demonstrated here provides a new opportunity for the development of phosphine free transition-metal catalysis. PMID- 19123227 TI - On the possibility of conversion of alcohols to ketones and aldehydes by phosphinoboranes R2PBR'R'': a computational study. AB - Can phosphinoboranes promote hydrogenation of carbonyl moieties? By means of B3LYP and MPW1K density functional calculations the likelihood of the oxidation of alcohols by phosphinoboranes R(2)PBR'(2) (1) was explored. As a proof-of principle, a theoretical study that tests the reversibility of the alcohol oxidation is reported. The potential of 1 as a metal-free hydrogenation mediator is discussed for a series of hydrogen sources such as primary and secondary alcohols. PMID- 19123228 TI - Asymmetric gold-catalyzed hydroarylation/cyclization reactions. PMID- 19123229 TI - Catalyzed vinylogous Mukaiyama aldol reactions with controlled enantio- and diastereoselectivities. AB - In control: A new catalytic vinylogous Mukaiyama aldol reaction provides products with high diastereo- and enantioselectivities (up to 99 % de and ee; see scheme). The relative and absolute stereochemistry of a representative product was rigorously assigned by NMR and CD spectroscopies (measured and calculated), X-ray diffraction, and quantum-chemical calculations. PMID- 19123231 TI - Spindle misorientation in tumors from APC(min/+) mice. AB - The adenomatous polyposis coli (APC) tumor suppressor gene is mutated in the majority of colon cancers, and its mutation may initiate cancer by multiple mechanisms. Recently, abnormal mitotic spindle orientation was shown in normal appearing tissues from mice heterozygous for APC mutation. To determine the effect of APC mutation on spindle orientation in tumors, and to better understand its mechanism, we measured mitotic spindle orientation in intestinal tumors from APC mutant mice, with three-dimensionally reconstructed confocal stacks of microtubule immunofluorescence images. We found spindle angles were increased in crypts heterozygous for the APC(min) mutation, and further increased in tumors. Astral microtubules of these spindles were clearly evident, suggesting astral microtubule loss is not a major mechanism of spindle misorientation in intestinal cells lacking wild-type APC. beta-Catenin staining was markedly abnormal in crypts and tumors from the mutant mice, suggesting a possible role in spindle orientation. Spindle angles in colon tumors with wild-type APC were equivalent to those in wild-type mice, showing that spindle misorientation is specific to APC mutation and not a general feature of tumors. We suggest spindle misorientation may contribute to the loss of normal tissue organization during tumor formation and could offer new insights into early carcinogenic events. PMID- 19123230 TI - The MT pool size ratio and the DTI radial diffusivity may reflect the myelination in shiverer and control mice. AB - A quantitative magnetization transfer (qMT) technique was employed to quantify the ratio of the sizes of the bound and free water proton pools in ex vivo mouse brains. The goal was to determine the pool size ratio sensitivity to myelin. Fixed brains from both shiverer mice and control littermates were imaged. The pool size ratio in the corpus callosum of shiverer mice was substantially lower than that in the control mice, while there was no distinguishable difference in the pool size ratio in the gray matter. These results correlate with diffusion tensor imaging (DTI) derived radial diffusivity which previously was shown to reflect myelin integrity in this animal model. Histological study reveals the presence of myelin in control mice white matter and the absence of myelin in shiverer mice white matter, supporting the qMT and DTI results. Our findings support the view that qMT may be used for estimating myelin integrity. PMID- 19123233 TI - Therapy with religious couples. AB - Although 95% of married couples identify with a particular religion, there is great variation in how couples rely on their religion to define or structure their relationship. Various denominations will imply particular "rules" or will shape how the couple deals with interpersonal and family challenges, such as sexuality, parenting, and power. In this article, we review couple relationships within a religious context and advance several treatment principles for treating religious couples. We present a clinical case to illustrate marital therapy with a religious couple, with an Adlerian context. PMID- 19123232 TI - Effects of multiple chondroitinase ABC applications on tissue engineered articular cartilage. AB - Increasing tensile properties and collagen content is a recognized need in articular cartilage tissue engineering. This study tested the hypothesis that multiple applications of chondroitinase ABC (C-ABC), a glycosaminoglycan (GAG) degrading enzyme, could increase construct tensile properties in a scaffold-less approach for articular cartilage tissue engineering. Developing constructs were treated with C-ABC at 2 weeks, 4 weeks, or both 2 and 4 weeks. At 4 and 6 weeks, construct sulfated GAG composition, collagen composition, and compressive and tensile biomechanical properties were assessed, along with immunohistochemistry (IHC) for collagens type I, II, and VI, and the proteoglycan decorin. At 6 weeks, the tensile modulus and ultimate tensile strength of the group treated at both 2 and 4 weeks were significantly increased over controls by 78% and 64%, reaching values of 3.4 and 1.4 MPa, respectively. Collagen concentration also increased 43%. Further, groups treated at either 2 weeks or 4 weeks alone also had increased tensile stiffness compared to controls. Surprisingly, though GAG was depleted in the treated groups, by 6 weeks there were no significant differences in compressive stiffness. IHC showed abundant collagen type II and VI in all groups, with no collagen type I. Further, decorin staining was reduced following C-ABC treatment, but returned during subsequent culture. The results support the use of C-ABC in cartilage tissue engineering for increasing tensile properties. PMID- 19123234 TI - Spiritually oriented psychodynamic psychotherapy. AB - Spiritually oriented psychodynamic psychotherapy pays particular attention to the roles that religious and spiritual beliefs, practices, and experiences play in the psychological life of the client. Contemporary psychoanalytic theorists offer multiple approaches to understand the functions of religious experience. Spirituality provides a means to address existential issues and provide a context to form personal meaning. Religious narratives present schemas of relationship and models of experiences salient to mental health, such as hope. God images or other symbolic representations of the transcendent have the power to evoke emotions, which in turn, influence motivation and behavior. While employing theories and techniques derived from psychodynamic psychotherapy, this therapeutic approach encourages the analysis of the functions religion and spirituality serve, while respecting the client's act of believing in faith. Psychotherapists address a client's spirituality by exploring the psychological meaning of such personal commitments and experiences and refrain from entering into discussion of faith claims. PMID- 19123235 TI - Neural organization of first optic neuropils in the littoral crab Hemigrapsus oregonensis and the semiterrestrial species Chasmagnathus granulatus. AB - Crustaceans are among the most extensively distributed arthropods, occupying many ecologies and manifesting a great variety of compound eye optics; but in comparison with insects, relatively little is known about the organization and neuronal morphologies of their underlying optic neuropils. Most studies, which have been limited to descriptions of the first neuropil--the lamina--suggest that different species have approximately comparable cell types. However, such studies have been limited with regard to the types of neurons they identify and most omit their topographic relationships. It is also uncertain whether similarities, such as they are, are independent of visual ecologies. The present account describes and compares the morphologies and dispositions of monopolar and other efferent neurons as well as the organization of tangential and smaller centrifugal neurons in two grapsoid crabs, one from the South Atlantic, the other from the North Pacific. Because these species occupy significantly disparate ecologies we ask whether this might be reflected in differences of cell arrangements within the most peripheral levels of the visual system. The present study identifies such differences with respect to the organization of centrifugal neurons to the lamina. We also identify in both species neurons in the lamina that have hitherto not been identified in crustaceans and we draw specific comparisons between the layered organization of the grapsoid lamina and layered laminas of insects. PMID- 19123236 TI - Hepatic resection for hepatocellular carcinoma in the elderly. AB - BACKGROUND AND OBJECTIVES: Aging of the population has significantly increased the number of elderly patients undergoing surgery for hepatocellular carcinoma (HCC). We aimed to compare the results of hepatectomy for HCC in patients >/=70 years old with those for younger patients. METHODS: Clinicopathological data and outcomes for 155 elderly patients and 333 younger patients with HCC who underwent hepatectomy between 1992 and 2007 were retrospectively compared. RESULTS: The elderly group had a higher incidence of associated diseases, but had better preoperative liver function. Although postoperative delirium was more common in the elderly group, there were no significant differences between the two groups with regard to operative morbidity, hospital death, disease-free survival, and overall survival. The overall recurrence rate was significantly higher in the elderly patients with alcohol abuse than in younger patients with alcohol abuse. Multivariate analysis revealed that preoperative alcohol abuse was a prognostic factor for elderly patients. CONCLUSIONS: The short-term and long-term outcomes of surgery for HCC were similar in elderly and younger patients. Elderly patients with preoperative alcohol abuse should be followed closely, even after R0 surgery, because alcohol abuse is strongly correlated with postoperative recurrence and worse survival. PMID- 19123237 TI - Aerobic fitness is associated with hippocampal volume in elderly humans. AB - Deterioration of the hippocampus occurs in elderly individuals with and without dementia, yet individual variation exists in the degree and rate of hippocampal decay. Determining the factors that influence individual variation in the magnitude and rate of hippocampal decay may help promote lifestyle changes that prevent such deterioration from taking place. Aerobic fitness and exercise are effective at preventing cortical decay and cognitive impairment in older adults and epidemiological studies suggest that physical activity can reduce the risk for developing dementia. However, the relationship between aerobic fitness and hippocampal volume in elderly humans is unknown. In this study, we investigated whether individuals with higher levels of aerobic fitness displayed greater volume of the hippocampus and better spatial memory performance than individuals with lower fitness levels. Furthermore, in exploratory analyses, we assessed whether hippocampal volume mediated the relationship between fitness and spatial memory. Using a region-of-interest analysis on magnetic resonance images in 165 nondemented older adults, we found a triple association such that higher fitness levels were associated with larger left and right hippocampi after controlling for age, sex, and years of education, and larger hippocampi and higher fitness levels were correlated with better spatial memory performance. Furthermore, we demonstrated that hippocampal volume partially mediated the relationship between higher fitness levels and enhanced spatial memory. Our results clearly indicate that higher levels of aerobic fitness are associated with increased hippocampal volume in older humans, which translates to better memory function. PMID- 19123238 TI - Topographic commissural and descending projections of the habenula in the rat. AB - The habenular complex of the epithalamus connects the limbic basal forebrain with numerous neuromodulatory centers in the midbrain. The habenula consists of the medial and lateral nuclei, each of which is speculated to contain multiple subdivisions. Such anatomical arrangements raise the possibility that the habenula accounts for multiple channels of information flow from the limbic forebrain to the midbrain. For understanding whether and, if so, how the multiple streams are organized via the habenula, knowledge of the precise input-output connectivity of each habenular subdivision is essential. In the present study, biotinylated dextran amine and cholera toxin subunit B were used to delineate the differential outputs of various subregions of the medial and lateral nuclei of the habenula in the rat. Both anterograde and retrograde tracing uncovered a heavy commissural connection between the two habenulae on the ipsilateral and contralateral sides. The commissural projection arises primarily in the lateral nucleus and exhibits a fine topography in that a local commissural efferent terminates primarily in the corresponding subregion on the contralateral side. The subregions of the medial and lateral nuclei also give rise to distinct projections to midbrain areas such as the interpeduncular nucleus, the median/paramedian nuclei, and the central gray. These projections produce terminal fields centered in different areas of the targets, supporting the topographically organized descending projections from the habenula. These data together support the organization of multiple channels in the habenula that convey parallel streams of information to the contralateral habenula, midbrain, and brainstem. PMID- 19123239 TI - A light and electron microscopic study on the organization of the testis and the semicystic spermatogenesis of the genus Scorpaena (Teleostei, Scorpaenidae). AB - The testicular organization and semicystic spermatogenesis of Scorpaena porcus and Scorpaena scrofa are analyzed by means of optic and electron microscopy and immunohistochemical techniques. The testicular structure of S. porcus and S. scrofa belongs to the unrestricted spermatogonial type, but has typical features of the restricted type. Moreover, the structure presents an epithelioid arrangement of Sertoli and germ cells rather than the germinal epithelium that appears in the majority of teleosts. After the cysts open, Sertoli cells hypertrophy and remain on the basement membrane, linked by interdigitations and tight junctions and bordering the lumen of the lobule, which at this moment works as an efferent duct. Secretions of Sertoli cells usually function in the nutrition of germ cells, and they seem to contribute in it even in this kind of spermatogenesis in which the free lumen spermatids do not have any connection with Sertoli cells. In addition, Sertoli cells can divide after the cysts have broken apart and hypertrophied, suggesting that they are still important for the final maturation of spermatozoa and seminal fluid formation. PMID- 19123240 TI - Forelimb indicators of prey-size preference in the Felidae. AB - The forelimbs, along with the crania, are an essential part of the prey-killing apparatus in cats. Linear morphometrics of the forelimbs were used to determine the morphological differences between felids that specialize on large prey, small prey, or mixed prey. We also compared the scaling of felid forelimbs to those of canids to test whether prey capture strategies affect forelimb scaling. Results suggest that large prey specialists have relatively robust forelimbs when compared with smaller prey specialists. This includes relatively more robust humeri and radii, relatively larger distal ends of the humerus, and relatively larger articular areas of the humerus and radius. Large prey specialists also had relatively longer olecranon processes of the ulna and wider proximal paws. These characters are all important for subduing large prey while the cat positions itself for the killing bite. Small prey specialists have relatively longer distal limb elements for swift prey capture, and mixed prey specialists had intermediate values with relatively more robust metacarpals. Arboreal felids also had more robust limbs. They had relatively longer proximal phalanges for better grip while climbing, and a relatively short brachial index (radius to humerus ratio). Additionally, we found that felids and canids differ in forelimb scaling, which emphasizes the dual use of forelimbs for locomotion and prey capture in felids. This morphometric technique worked well to separate prey-size preference in felids, but did not work as well to separate locomotor groups, as scansorial and terrestrial felids were not clearly distinguished. PMID- 19123241 TI - Morphological specializations of the buccal cavity in relation to the food and feeding habit of a carp Cirrhinus mrigala: a scanning electron microscopic investigation. AB - The buccal cavity of an herbivorous fish, Cirrhinus mrigala, was investigated by scanning electron microscopy to determine its surface ultrastructure. The buccal cavity shows significant adaptive modifications in relation to food and feeding ecology of the fish. The buccal cavity of the fish is of modest size and limited capacity, which is considered an adaptation with respect to the small-sized food items primarily consumed by the fish that could be accommodated in a small space. Modification of surface epithelial cells, on the upper jaw, into characteristic structures-the unculi-is considered an adaptation to browse or scrap, to grasp food materials, e.g., algal felts, and to protect the epithelial surface against abrasions, likely to occur during their characteristic feeding behavior. Differentiation of the highly specialized lamellar organ on the anterior region of the palate could be an adaptation playing a significant role in the selection, retention, and sorting out of palatable food particles from the unpalatable items ingested by the fish. The filamentous epithelial projections and the lingulate epithelial projections on the palatal organ in the posterior region of the palate are considered to serve a critical function in final selection, handling, maneuvering, and propelling the food particles toward the esophagus. The abundance of different categories of taste buds in the buccal cavity suggests that gustation is well developed and the fish is highly responsive in the evaluation and the selection of the preferred palatable food items. The secretions of mucous cells in the buccal cavity are associated with multiple functions-particle entrapment, lubrication of the buccal epithelium and food particles to assist smooth passage of food, and to protect the epithelium from possible abrasion. These morphological characteristics ensure efficient working of the buccal cavity in the assessment of the quality and palatability of ingested food, their retention and transport toward the esophagus. Such an adaptation may be essential in conducting the function most basic to the survival of the individuals and species-feeding. PMID- 19123242 TI - The comparative morphology of pit organs in elasmobranchs. AB - The pit organs of elasmobranchs (sharks, skates and rays) are free neuromasts of the mechanosensory lateral line system. Pit organs, however, appear to have some structural differences from the free neuromasts of bony fishes and amphibians. In this study, the morphology of pit organs was investigated by scanning electron microscopy in six shark and three ray species. In each species, pit organs contained typical lateral line hair cells with apical stereovilli of different lengths arranged in an "organ-pipe" configuration. Supporting cells also bore numerous apical microvilli taller than those observed in other vertebrate lateral line organs. Pit organs were either covered by overlapping denticles, located in open grooves bordered by denticles, or in grooves without associated denticles. The possible functional implications of these morphological features, including modification of water flow and sensory filtering properties, are discussed. PMID- 19123243 TI - Developmental stages until hatching of the Lake Victoria cichlid Haplochromis piceatus (Teleostei: Cichlidae). AB - Because little is known about embryonic developmental stages in any haplochromine cichlid, we describe here a series of normal stages of the Lake Victoria cichlid Haplochromis piceatus. We collected 273 embryos and scored them for 47 morphological characters. The result was an illustrated series of 12 stages from embryonic shield until hatching based on live, fixed, and histological material. We defined each stage according to a single "key" character that applied to all embryos of that stage. Other characters forming part of the stage descriptions were not necessarily present in all embryos of a stage. We compare our findings with published studies of other freshwater teleosts and find wide variation in staging systems. Our data will form a baseline for further research on cichlid development. PMID- 19123244 TI - Neuroanatomy of the calf brain as revealed by high-resolution magnetic resonance imaging. AB - Here, we want to assess the benefit of high-resolution and high-contrast magnetic resonance imaging (MRI) for detailed documentation of internal brain morphology in formalin-fixed whole head specimens of the full-term calf brain (Bos taurus). Imaging was performed on a Siemens 1.5 T scanner. Optimum contrast was achieved using a 3D sequence with a flip angle of 30 degrees , repetition time (TR) of 20 ms, echo time (TE) of 6.8 ms, and an interpolated matrix of 1024 x 1024. In plane resolution was 0.25 mm. Computer-generated three-dimensional images were reconstructed from the original scans in the coronal plane. This study shows that MRI is capable to identify delicate structures in immature brain specimens. The use of MRI in comparative morphology facilitates the examination of series of brains or brain samples in a reasonable time. The comprehensive description of species- and group-specific brain features in MRI scans of Bos taurus will complement existing data for diagnostic imaging and neuromorphological research, in general, as well as for phylogenetic reconstructions. PMID- 19123245 TI - Ultrastructural and biochemical seasonal changes in epididymal corpus and cauda of viscacha (Lagostomus maximus maximus). AB - The reproductive and adaptative behavior of wild rodents is synchronized primarily by the photoperiod. The viscacha, a South American rodent of nocturnal habits and seasonal reproduction is photoperiod-dependent and its reproductive behavior is regulated by the retinohypothalamic-pituitary pineal axis. Adult males exhibit an annual reproductive cycle with periods of maximum gonadal activity (summer-early autumn) and gonadal regression (winter). The corpus and the cauda, the most sensitive segments of the epididymis to changes induced by the photoperiod, were analyzed using electron microscopy and enzymatic biochemistry. During gonadal regression, principal and clear cells showed signs of involution with respect to the activity period. These were characterized by more irregular nuclei, smaller cytoplasms, large vacuoles, altered mitochondria, and glycogen deposits. All cellular populations of the epididymal epithelium in regression presented abundant lysosome-like dense bodies during the active period. In addition, we measured the activity of four acid glycosidases in the cauda epididymis along the reproductive cycle. N-acetyl-beta-D-glucosaminidase (NAG), an enzyme that degrades endocytosed substances from the epididymal lumen, increased significantly during gonadal regression relative to the active period. These results demonstrate that the viscacha epididymis exhibits significant ultrastructural and biochemical changes during the reproductive cycle. We demonstrate that during regression, melatonin secretion in viscacha increases. This study shows that the epididymal epithelium is reduced. Thus, we postulate that the changes observed in the epididymis are modulated by pineal melatonin. Despite these changes, the epididymis might maintain a microenvironment suitable for the survival of stored spermatozoa. PMID- 19123246 TI - Quantitative estimates of visual performance features in fossil birds. AB - Eyeball structures such as the lens diameter (LD) and axial length are generally assumed to be highly correlated with optically meaningful parameters. However, these optical constraints on eyeball macroanatomy have never been tested explicitly. Tradeoffs between benefits of improved visual performance and cost of adaptation from an increase of tissue production predict that when eyeball size increases, optical parameters such as posterior nodal distance and maximum entrance pupil diameter should increase isometrically with eyeball axial length and LD, respectively. Here I show quantitatively that the interspecific allometry of the avian eye largely follows this predicted isometry. Additionally, I elaborate a method to estimate optically significant eyeball soft-tissue dimensions from scleral ring and orbit morphology based on analyses of interspecific allometry in Aves. The stringent correlations between avian eyeball morphology and optical function render this system ideal for the analysis of form function relationships and allow for an accurate estimate of optically significant eyeball soft-tissue dimensions such as diameter, axial length, and LD in fossil species. PMID- 19123247 TI - A novel membrane specialization in the sperm tail of bug insects (Heteroptera). AB - The sperm tail of bug insects has 9 + 9 + 2 flagellar axonemes and two mitochondrial derivatives showing two to three crystalline inclusions in their matrix. During spermiogenesis, the axoneme is surrounded by a membrane cistern which, at sperm maturity, reduces to two short cisterns on the opposite sides of the axoneme adhering to the mitochondrial derivatives. Filamentous bridges connect the intertubular material of the axoneme to these cisterns. Such bridges, which represent a peculiar feature of bug insects, are resistant to detergent treatment, whereas part of the intertubular material and the inner content of microtubular doublets are affected by the treatment. After freeze-fracture replicas, at the insertion of the bridges to the cisternal membrane, the P-face of this membrane shows a characteristic ribbon consisting of four rows of 11 +/- 1 nm staggered intramembrane particles, 13 +/- 2 nm apart along each row. The bridges could be able to maintain the axoneme in the proper position during flagellar beating avoiding distortion affecting sperm motility. PMID- 19123248 TI - Developmental changes in the ultrastructure of the lamprey lateral line nerve during metamorphosis. AB - The ultrastructure of the trunk lateral line nerve of larval and adult lampreys was studied with transmission electron microscopy. We confirmed that lampreys' lateral line nerve lacks myelin. Nevertheless, all axons were wrapped by Schwann cell processes. In the larval nerve, gaps between Schwann cells were observed, where the axolemma was covered only by a basal lamina, indicating an earlier developmental stage. In the adult nerve, glial (Schwann cell) ensheathment was mostly complete. Additionally, we observed variable ratios of axons to Schwann cells in larval and adult preparations. In the larval nerve, smaller axons were wrapped by one Schwann cell. Occasionally, a single Schwann cell surrounded two axons. Larger axons were associated with two to five Schwann cells. In the adult nerve, smaller axons were surrounded by one, but larger axons by three to eight Schwann cells. The larval epineurium contained large adipose cells, separated from each other by single fibroblast processes. This layer of adipose tissue was reduced in adult preparation. The larval perineurium was thin, and the fibroblasts, containing large amounts of glycogen granules, were arranged loosely. The adult perineurium was thicker, consisting of at least three layers of fibroblasts separated by collagen fibrils. The larval and adult endoneurium contained collagen fibrils oriented orthogonally to each other. Both larval and adult lateral line nerves possessed a number of putative fascicles weakly defined by a thin layer of perineurial fibroblasts. These results indicate that after a prolonged larval stage, the lamprey lateral line nerve is subjected to additional maturation processes during metamorphosis. PMID- 19123249 TI - Effects of parity on pelvic size and shape dimorphism in Mus. AB - The pelvis is a sexually dimorphic structure and although the causes of that dimorphism have long been studied, relatively little is known regarding the effects of partuitive events on the magnitude of that dimorphism. Here, we use a sample of Mus musculus domesticus to contrast dimorphism in body length and os coxae size and shape between males and parous and nulliparous females. We also test for correlations between relative litter size (L/M) and relative offspring size (O/M) with body length and os coxae size and shape in parous females. Males had greater body length than nulliparous females but were not different from parous females. Females as a whole had the largest os coxae, with parous females having the largest and males the smallest. Os coxae shape was also significantly different between groups and was most divergent between parous females and males than between nulliparous females and males. Os coxae shape differences between females are associated with differences in body length between females and O/M is correlated with os coxae shape in parous females such that females with the largest offspring have the most divergent shapes along the relative warp one axis. Pelvic shape differences between males and females were consistent with previous findings in other taxa which identify the pubo-ischial complex as the primary region of dimorphism. PMID- 19123251 TI - Does the cingulate cortex contribute to spatial conditional associative learning in the rat? AB - Rats with lesions to the anterior or posterior (retrosplenial) region of the cingulate cortex and rats with lesions that included both the anterior and posterior cingulate cortex were tested on a visual-spatial conditional task in which they had to learn to approach one of the two objects depending on the spatial context within which they were embedded. Lesions restricted to either the anterior or the retrosplenial cingulate region did not impair learning of this task which is known to be very sensitive to the effects of hippocampal lesions. Complete lesions of the cingulate cortex gave rise to only a minor retardation in learning. In contrast, lesions to the retrosplenial cortex impaired performance on a spatial navigation task and the classic radial maze. These results suggest that the retrosplenial portion of the cingulate region forms part of a hippocampal circuit underlying learning about spatial responses. The dissociation between the effects of lesions of the cingulate region on different classes of behavior known to be associated with hippocampal function suggests that, although this neural structure does play a role in an extended hippocampal circuit underlying spatial learning, its role in such learning may be a selective one. PMID- 19123250 TI - Correlates of reward-predictive value in learning-related hippocampal neural activity. AB - Temporal difference learning (TD) is a popular algorithm in machine learning. Two learning signals that are derived from this algorithm, the predictive value and the prediction error, have been shown to explain changes in neural activity and behavior during learning across species. Here, the predictive value signal is used to explain the time course of learning-related changes in the activity of hippocampal neurons in monkeys performing an associative learning task. The TD algorithm serves as the centerpiece of a joint probability model for the learning related neural activity and the behavioral responses recorded during the task. The neural component of the model consists of spiking neurons that compete and learn the reward-predictive value of task-relevant input signals. The predictive value signaled by these neurons influences the behavioral response generated by a stochastic decision stage, which constitutes the behavioral component of the model. It is shown that the time course of the changes in neural activity and behavioral performance generated by the model exhibits key features of the experimental data. The results suggest that information about correct associations may be expressed in the hippocampus before it is detected in the behavior of a subject. In this way, the hippocampus may be among the earliest brain areas to express learning and drive the behavioral changes associated with learning. Correlates of reward-predictive value may be expressed in the hippocampus through rate remapping within spatial memory representations, they may represent reward-related aspects of a declarative or explicit relational memory representation of task contingencies, or they may correspond to reward related components of episodic memory representations. These potential functions are discussed in connection with hippocampal cell assembly sequences and their reverse reactivation during the awake state. The results provide further support for the proposal that neural processes underlying learning may be implementing a temporal difference-like algorithm. PMID- 19123252 TI - Age-dependent enhancement of inhibitory synaptic transmission in CA1 pyramidal neurons via GluR5 kainate receptors. AB - Changes in hippocampal synaptic networks during aging may contribute to age dependent compromise of cognitive functions such as learning and memory. Previous studies have demonstrated that GABAergic synaptic transmission exhibits age dependent changes. To better understand such age-dependent changes of GABAergic synaptic inhibition, we performed whole-cell recordings from pyramidal cells in the CA1 area of acute hippocampal slices on aged (24-26 months old) and young (2 4 months old) Brown-Norway rats. We found that the frequency and amplitude of spontaneous inhibitory postsynaptic current (IPSCs) were significantly increased in aged rats, but the frequency and amplitude of mIPSCs were decreased. Furthermore, the regulation of GABAergic synaptic transmission by GluR5 containing kainate receptors was enhanced in aged rats, which was revealed by using LY382884 (a GluR5 kainate receptor antagonist) and ATPA (a GluR5 kainate receptor agonist). Moreover, we demonstrated that vesicular glutamate transporters are involved in the kainate receptor dependent regulation of sIPSCs. Taken together, these results suggest that GABAergic synaptic transmission is potentiated in aged rats, and GluR5 containing kainate receptors regulate the inhibitory synaptic transmission through endogenous glutamate. These alterations of GABAergic input with aging could contribute to age-dependent cognitive decline. PMID- 19123253 TI - Prenatal screening for Down syndrome: women's involvement in decision-making and their attitudes to screening. AB - OBJECTIVE: To investigate pregnant women's decisions about and attitudes towards Down syndrome screening, i.e. ultrasound at 11-14 weeks, Maternal Serum Markers (MSM) at 11-14 or 15-17 weeks and possibly invasive testing. METHOD: Women having given birth to a non-affected child were asked to fill in a self-administered questionnaire during their stay at the maternity unit. In order to characterize women's decision-making behaviour, a hierarchical cluster analysis was conducted and logistic regressions were performed. RESULTS: Four hundred women were invited to participate in the study, and 391 returned the questionnaire. Both ultrasound and biochemical screening had been proposed to 88.3% of the women. Three clusters of women who were offered ultrasound and MSM were identified. Two clusters (52% and 42% of women) differed in active versus passive involvement in decision making. Passively involved women frequently reported unawareness of the possibility of having to make decisions about invasive testing and/or termination of pregnancy. The third cluster (6% of women) consisted of women who declined MSM. Most of the women showed a preference for first-trimester screening, but actively involved women were willing to pay more for MSM. CONCLUSION: Providing information about the sequence of decision possibly involved in screening could contribute to better informed decisions. PMID- 19123254 TI - HIV doctors may treat all infected, adding thousands. PMID- 19123255 TI - A new tool to predict your risk of diabetes. Assess your probability with the Diabetes Risk Calculator. PMID- 19123256 TI - Improvements in heart attack prevention and treatment. New methods save lives both before and after heart attacks occur. PMID- 19123257 TI - Medication and talk therapy help prevent depression after a stroke. Antidepressants and problem-solving therapy can prevent or delay the onset of post-stroke depression. PMID- 19123258 TI - Exercise improves cancer survival rates in men. Walking or cycling 30 minutes a day may increase your chance of surviving cancer by 33 percent. PMID- 19123259 TI - I'm a 65 year old woman who was recently diagnosed with osteopenia. How is this similar to osteoporosis, and how is it treated? PMID- 19123260 TI - Can eating processed foods increase my risk of colon cancer? Are there foods that can reduce my risk? PMID- 19123261 TI - When are heart palpitations a concern? PMID- 19123262 TI - The human gene map for performance and health-related fitness phenotypes: the 2006-2007 update. AB - This update of the human gene map for physical performance and health-related fitness phenotypes covers the research advances reported in 2006 and 2007. The genes and markers with evidence of association or linkage with a performance or a fitness phenotype in sedentary or active people, in responses to acute exercise, or for training-induced adaptations are positioned on the map of all autosomes and sex chromosomes. Negative studies are reviewed, but a gene or a locus must be supported by at least one positive study before being inserted on the map. A brief discussion on the nature of the evidence and on what to look for in assessing human genetic studies of relevance to fitness and performance is offered in the introduction, followed by a review of all studies published in 2006 and 2007. The findings from these new studies are added to the appropriate tables that are designed to serve as the cumulative summary of all publications with positive genetic associations available to date for a given phenotype and study design. The fitness and performance map now includes 214 autosomal gene entries and quantitative trait loci plus seven others on the X chromosome. Moreover, there are 18 mitochondrial genes that have been shown to influence fitness and performance phenotypes. Thus,the map is growing in complexity. Although the map is exhaustive for currently published accounts of genes and exercise associations and linkages, there are undoubtedly many more gene-exercise interaction effects that have not even been considered thus far. Finally, it should be appreciated that most studies reported to date are based on small sample sizes and cannot therefore provide definitive evidence that DNA sequence variants in a given gene are reliably associated with human variation in fitness and performance traits. PMID- 19123263 TI - Dr. Michael DeBakey's contributions to emergency medicine and trauma care. PMID- 19123264 TI - Update on emerging infections: news from the Centers for Disease Control and Prevention. False-positive oral fluid rapid HIV tests--New York City, 2005-2008. PMID- 19123266 TI - Beyond the crunch. PMID- 19123265 TI - An international survey: Public awareness and perception of sepsis. AB - BACKGROUND: Sepsis is a common cause of death throughout the world. Early treatment improves outcome; however, treatment may be delayed if the patient does not present himself/herself for medical care until late in the disease process. Lack of knowledge about the syndrome may contribute to delay in presenting for medical care. However, we need to acknowledge the complexity of sepsis. General awareness of sepsis by the public may increase political pressure for research funding. Increased public awareness of acute myocardial infarction has contributed to reduced mortality over the last 50 yrs. This example provides a rationale for future efforts to increase the public awareness of sepsis. OBJECTIVE: The survey was designed to gain insight into public perceptions and attitudes regarding sepsis. DESIGN: Prospective, international survey performed using structured telephone interviews. SUBJECTS: A total of 6021 interviewees, 5021 in Europe and 1000 in the United States. MEASUREMENTS AND MAIN RESULTS: In Italy, Spain, the United Kingdom, France and the United States, a mean of 88% of interviewees had never heard of the term "sepsis". In Germany 53% of people knew the word sepsis. In Italy, Spain, United Kingdom, France, and United States, of people who recognized the term sepsis, 58% did not recognize that sepsis is a leading cause of death. CONCLUSIONS: There is poor public awareness about the existence of a syndrome known as sepsis. Results of this questionnaire underscore the challenges in early management and treatment of infected patients at risk for developing sepsis syndrome. PMID- 19123267 TI - From surgical to molecular scalpel: ERJ lung cancer series for 2009. PMID- 19123268 TI - Expression of RAC2 in endothelial cells is required for the postnatal neovascular response. AB - Herein, we describe an obligate role for the hematopoietic specific GTPase, RAC2 in endothelial integrin signaling and the postnatal neovascularization response in vivo. Using a Rac2 knockout mouse model, we discovered that despite the presence of both RAC1 and RAC2 protein in endothelial cells, RAC2 is obligately required for the postnatal neovascular response and alphavbeta3/ alpha4beta1/alpha5beta1 integrin-directed migration on vitronectin, H296 and CH271, fibronectin fragments, respectively. The molecular basis for RAC2 specificity was explored. A genetic analysis of Syk -/+ or Syk-/+;Rac2 -/+ mice revealed that SYK kinase is required for the integrin induced activation of RAC2. The analysis of endothelial cells from Rac2-/+ versus Syk-/+;Rac2-/+ mice provided genetic evidence that SYK-RAC2 signaling axis regulates integrin (alphavbeta3, alpha4beta1 and alpha5beta1) dependent migration. Our results provide evidence that a specific region of the nonreceptor protein tyrosine kinase, SYK, the B linker region containing Y342 and Y346 is required for SYK's regulation of RAC2 and integrin dependent migration. Moreover, the capacity of mice to vascularize the ischemic hindlimb following femoral artery ligation or matrigel plugs was markedly reduced in mice homozygous deficient for the Rac2 gene. These findings identify a novel signaling axis for the induction and potential modulation of postnatal angiogenesis. PMID- 19123270 TI - In memoriam: Roger A. Davis (1945-2008). PMID- 19123271 TI - Gilead AIDS pill effective in study with weekends off. PMID- 19123269 TI - Empirically controlled mapping of the Caenorhabditis elegans protein-protein interactome network. AB - To provide accurate biological hypotheses and elucidate global properties of cellular networks, systematic identification of protein-protein interactions must meet high quality standards.We present an expanded C. elegans protein-protein interaction network, or 'interactome' map, derived from testing a matrix of approximately 10,000 x approximately 10,000 proteins using a highly specific, high-throughput yeast two-hybrid system. Through a new empirical quality control framework, we show that the resulting data set (Worm Interactome 2007, or WI 2007) was similar in quality to low-throughput data curated from the literature. We filtered previous interaction data sets and integrated them with WI-2007 to generate a high-confidence consolidated map (Worm Interactome version 8, or WI8). This work allowed us to estimate the size of the worm interactome at approximately 116,000 interactions. Comparison with other types of functional genomic data shows the complementarity of distinct experimental approaches in predicting different functional relationships between genes or proteins PMID- 19123272 TI - Male circumcision booms in Uganda. PMID- 19123273 TI - Scientists to test tuberculosis vaccine in Cape Town. PMID- 19123274 TI - ASEAN-UN study find migrants vulnerable to AIDS. PMID- 19123276 TI - Editorial comment: we're doing something right! PMID- 19123275 TI - Decrease in the proportion of injecting drug use-related HIV/AIDS in Massachusetts, New York, Connecticut, and Rhode Island. AB - Injection drug use (IDU) has been a route for HIV transmission since the beginning of the epidemic. This review focuses on an apparent decrease in the proportion of IDU-related HIV/AIDS in Massachusetts, New York, Connecticut, and Rhode Island from 1999 to the present. Although there are differences in HIV/AIDS reporting requirements among these states, the dramatic reduction in the proportion of HIV and AIDS cases related to IDU is notable and is comparable to the 42% decline in HIV diagnoses among injection drug users reported by the CDC in other areas of the country. PMID- 19123277 TI - Lymphoma presenting as cranial nerve neuropathies in HIV-infected patients. PMID- 19123278 TI - HIV-associated pseudotumor cerebri: a case report and literature review. AB - Idiopathic intracranial hypertension is a cause of vision loss in HIV-positive patients. In many patients with controlled HIV disease, idiopathic intracranial hypertension develops without any other apparent cause. We report the case of an HIV-infected man in whom idiopathic intracranial hypertension developed, and despite therapy with repeated lumbar puncture, acetazolamide, and neurosurgical interventions, he did not regain his vision. PMID- 19123279 TI - Images in HIV/AIDS. HIV and viral hepatitis coinfection. PMID- 19123280 TI - Trends in HIV care, Part 1: new antiretrovirals, anal dysplasia, and reimbursement for diagnostic testing--a roundtable discussion. PMID- 19123282 TI - "There are huge anomalies in the capital that need to be addressed." Interview by Oona Mashta. PMID- 19123283 TI - [Trends in setting the ICD-11]. PMID- 19123284 TI - [Influence of the Japanese name change for schizophrenia on biased views of the disease]. PMID- 19123286 TI - Canguilhem and the history of biology. PMID- 19123285 TI - New lands, old ties and prostitution: a voiceless voice. PMID- 19123287 TI - The philosophy of science of Georges Canguilhem: a Transatlantic view. PMID- 19123288 TI - [Propriety of disease names translated in Japanese from English--social anxiety disorder, conduct disorder, and post-traumatic stress disorder]. PMID- 19123289 TI - [Confusion in disease names between depression and depressive disorder]. PMID- 19123290 TI - Adoption: a study of post-War child removal in New South Wales. PMID- 19123291 TI - The politics of famine in U.S. relations with Ethiopia, 1950-1970. PMID- 19123292 TI - Cesarean section. PMID- 19123293 TI - Neurology. PMID- 19123294 TI - [Extension of and confusion in concept of depression]. PMID- 19123295 TI - Cleaning a thousand waste-waters: water pollution in Finland, 1945-1970. PMID- 19123296 TI - Research reports on the diagnosis and detection; environment and infection; and evaluation of interventions on infectious diseases. PMID- 19123297 TI - Detection of a honeybee iflavirus with intermediate characteristics between kakugo virus and deformed wing virus. AB - Iflavirus RNA was detected in honeybee colonies displaying unduly aggressive behavior and with no evidence of morphological alterations. Sequence analysis of the RNA-dependent RNA polymerase (RdRp) revealed that the iflavirus strain was more similar (> 99% aa) to Deformed Wing Virus (DWV), that has been associated with morphological alterations in bees, rather than to the newly-described Kakugo Virus (KV) (about 95% aa), that has been associated with increased aggressiveness. Therefore, the iflavirus strain detected in the Italian hives genetically resembled DWV but was apparently associated with a KV-like phenotype. RT-PCR detected the iflavirus RNA in the abdomen of the workers, and only in one case was the virus detected in the head. No viral RNA was detected in the drones, a pattern of virus distribution across the honeybee casts that is in apparent conflict with the higher rates of infestation of drones by the mite Varroa distructor. The identification of a virus with apparently intermediate features between DWV and KV open new perspectives on the patho-biological role of iflaviruses in honeybees. PMID- 19123298 TI - A two-year prospective study of clinical criteria and polymerase chain reaction assay of cerebrospinal fluid for the diagnosis of viral infections of the central nervous system. AB - Cerebrospinal fluid specimens from 226 patients with suspected viral infections of the central nervous system (CNS) were tested by polymerase chain reaction (PCR) to identify the most frequent viruses involved in these infections. A positive PCR result was obtained in 18 patients (7 cases positive for herpes simplex viruses, 5 for enterovirus, 3 for JC virus, 2 for varicella-zoster virus, and 1 for Epstein-Barr virus). All patients with positive PCR results had a definite diagnosis of CNS viral infection. However, a negative result did not rule out the possibility of viral infection of the CNS. PMID- 19123299 TI - Prevalence of rubella and cytomegalovirus antibodies among pregnant women in northern Turkey. AB - Primary infections caused by rubella and cytomegalovirus (CMV) can lead to serious complications in pregnancy. Rubella and CMV screening of pregnant women is not routinely carried out in Turkey. The purpose of this study was to determine the prevalence of rubella and cytomegalovirus among pregnant women. The study was carried out in Samsun Maternity and Women's Disease and Pediatrics Hospital in Samsun province, Turkey. Between September 2004 and September 2005, 600 pregnant women aged 17-40 years were enrolled in this study. The results of the antenatal screening for rubella and CMV during the first trimester of pregnancy were evaluated. Anti-IgG against rubella seropositivity was found in 566 (94.3%) and rubella IgM seropositivity in 10 (1.7%). The positivity for anti CMV IgG antibody was found in 584 (97.3%), while 6 (1.0%) were positive for the anti-CMV IgM antibody. Pregnant women seronegative for rubella and CMV are susceptible to rubella and CMV primary infections. Preventive measures must be taken to decrease the mortality and morbidity related to congenital rubella and CMV infections. The rubella status should be investigated before pregnancy and seronegative females can be advised vaccination. PMID- 19123300 TI - Mosquito-based survey for the detection of flaviviruses and filarial nematodes in Aedes albopictus and other anthropophilic mosquitoes collected in northern Italy. AB - In order to optimize resources, an assay allowing the detection of several etiological agents would be useful. This paper describes a polymerase chain reaction (PCR)-based assay for the parallel detection of different pathogens in mosquitoes (Diptera: Culicidae). The assay combines the analysis of both RNA and DNA, hence allowing the detection of both flaviviruses (Flaviviridae: Flavivirus) and filarial nematodes (Spirurida: Onchocercidae). The sensitivity of diagnostic assay for both flavivirus and nematode infections was tested by means of spiked samples and resulted adequate for the purpose of screening mosquito samples. The assay was implemented to check pathogen infections in 637 mosquitoes field collected in Romagna, northern Italy. The integrity of RNA extracted from mosquito pools was checked by control amplifications targeting the 18 S rRNA of mosquitoes. Control amplifications were successful in 118 out of 119 pools. Flavivirus RNA was not found in any of these 118 pools, whereas DNA of Dirofilaria immitis (Leidy) was detected in one pool of Aedes albopictus (Skuse). PMID- 19123301 TI - Characterization of invasive serogroup Y meningococci in Italy: prevalence of ST 23 Complex/Cluster A3. AB - The percentage of Neisseria meningitidis serogroup Y isolated from patients with invasive meningococcal disease (IMD) in Italy has increased from 1998 to 2006. In this study, phenotypic features and genetic relatedness have been investigated in all serogroup Y meningococci isolated during that period. Multilocus sequence typing (MLST) identified the ST-23 complex/Cluster A3 as the major clonal complex in 88.8% of the strains. That complex included all strains belonging to the sequence type (ST) 23 isolated from 1998 to 2004, whereas the ST-3171 was prevalent among strains in the years 2005 and 2006. The STs 23 and 3171 differ for only one nucleotide in the phosphoglucomutase (pgm) housekeeping gene. Over 80% of serogroup Y ST-23 complex/Cluster A3 strains showed phenotype Y:14:NST and 85% of the latter resulted indistinguishable by pulsed-field gel electrophoresis analysis. In 2005, serogroup Y meningococci with decreased susceptibility to penicillin were isolated for the first time in Italy. In the following year, three of the seven strains showed this phenotype. The results of this study allow us to draw a profile of the molecular characteristics of invasive serogroup Y in Italy and will be helpful to monitor the spread of this serogroup in the next years. PMID- 19123302 TI - Genetic variability of genotype 1 hepatitis C virus isolates from Tunisian haemophiliacs. AB - This paper reports hepatitis C virus (HCV) prevalence, genotypes and phylogenetic characteristics in 95 haemophilic Tunisian patients. The studied population included 3 groups of patients according to their date of birth: before 1985 when inactivation procedures for clotting factors was introduced, between 1985 and 1994 when systematic anti-HCV screening of Tunisian blood donors was introduced and after this date. HCV infection was assessed by serological and molecular commercial tests. Genotypes were determined using the INNO-LiPA HCV test and by partial sequencing in the NS5b genomic region. Phylogenetic analyses were performed by comparing NS5b sequences of Tunisian haemophiliacs to published sequences. HCV infection was detected in 50.5% of cases with a significant decrease according to age. Subtype la was the most prevalent followed by subtype 1b (52.6% vs 44.7%); it was more frequent among haemophiliacs born before 1985. NS5b sequences were different from those obtained from non-haemophilic Tunisian patients and showed nucleic affiliation with HCV isolates from the USA. These findings suggest an infection through clotting factors imported to Tunisia and frequently manufactured from US blood donors. In contrast, subtype 1b showed approximately the same distribution among patients born before and after 1985; NS5b sequences from haemophiliacs were randomly distributed among other Tunisian sequences, favouring a transmission through cryoprecipitates prepared from Tunisian blood donors. PMID- 19123303 TI - Interaction of Aeromonas spp. human isolates with murine macrophages. AB - This study aimed to evaluate phagocytic and bactericidal activity of macrophages originated from murine J774 macrophage cell line against 26 strains of Aeromonas caviae, Aeromonas veronii biotype sobria and Aeromonas hydrophila isolated from human fecal specimens. The analysis of phagocytic index (PI) indicated these strains were phagocytosed in low numbers, below 1.1 x l0(-3) for 65% strains of A. veronii biotype sobria and A. hydrophila. The highest PI value (0.019) was noted in murine macrophages infected with 19% of A. caviae isolates. Analysis of the number of viable bacteria in phagocytes revealed that 31% of Aeromonas spp. strains could replicate for an initial 3 h after gentamicin treatment. Opsonisation of the bacteria with non-immune serum did not affect phagocytic activity and bacterial killing. Stimulation of macrophages with gamma-interferon (IFN-gamma) at a concentration of 10 IU/ml caused the highest increase in phagocytic activity but had no effect on the rate of bacteria elimination by murine macrophages. Aeromonas spp. isolates showed an invasive ability toward non phagocytic epithelial cells. These findings indicate that Aeromonas spp. strains have evolved mechanisms to avoid phagocytosis, suggesting that murine macrophages, even activated by IFN-gamma, have a limited ability to eliminate these bacteria, which may invade epithelial cells and spread in the illeum. PMID- 19123304 TI - Adhesive properties of environmental Vibrio alginolyticus strains to biotic and abiotic surfaces. AB - The ability of Vibrio alginolyticus strains isolated from a bathing and fishing area (Khenis, Centre of Tunisia) to adhere to both biotic and abiotic surfaces was evaluated in the present work. The biochemical, physiological and enzymatic activities of all strains was also investigated. Three morphotypes of V. alginolyticus were obtained on Congo red agar and only 14 strains produced black colonies. The majority of strains were able to degrade the skin mucus of both Sparus aurata and Dicentrarchus labrax fishes while the fish mucus preparation of these two specimens exhibits a high level of anti-V. alginolyticus strains. Adhesive properties were observed in 37.5% of the analyzed V. alginolyticus strains to Hep-2 cells and 50% to Caco-2 cells. All strains were able to form a purple pellicule on glass tube when they were stained with Crystal violet. Fifteen percent of V. alginolyticus strains (16/32) were strongly adhesive to polystyrene with a values ranging from 3.04 to 18.25 at 595 nm and only four strains were weak biofilm forming. V. alginolyticus bacterium possess a strong adhesive power to both biotic and inertes surfaces. These proprieties may allow to these strains to persist in this biotope in planctonic state or attached to both biotic and abiotic surfaces. PMID- 19123305 TI - Improving the efficiency and efficacy of pre-analytical and analytical work-flow of urine cultures with urinary flow cytometry. AB - Quantitative urine culture is one of the most frequently requested tests in microbiology laboratories. An automated system for screening purposes is strongly needed to save technical staff time and obtain rapid results. Our study investigated 1.047 urine samples collected from inpatients and outpatients with a commercial vacutainer system (Becton Dickinson, Milan, Italy) and compared a second-generation flow cytometry (Sysmex UF100, Dasit, Cornaredo, Italy) with standard urine culture tested on agar plated by means of 10 microliter loop. The specimens were screened and cultured on receipt. The results obtained with Sysmex UF-100 are very interesting, especially if this analyzer is used as a screening method for negative urine samples, and comparable to data obtained from culture examination. In fact, considering together bacteria and leukocyte count (> 4500 bacteria and/or > 50 leukocytes/microL) we obtained a negative predictive value of 99.5% in comparison with the standard culture method. The classical culture method needs 24 hours for a result, whereas the Sysmex UF-100 analyzer gives results in a few minutes, thus reducing the microbiology turn around time (TAT) with obvious benefits for patients and physicians. PMID- 19123306 TI - Dynamics of nasopharyngeal colonization by gram-negative rods in patients with resectable lung cancer during short-term hospitalization. AB - We determined the frequency of colonization of nasopharynx by Gram-negative rods in 63 patients with lung cancer undergoing thoracic surgery who routinely receive antimicrobial prophylaxis. Throat and nasal specimens were taken from each patient twice: on the day of hospital admission (examination I) followed by thoracic surgery and on the fourth day after thoracic surgery (examination II). The isolated strains were identified using API 20E or API 20NE microtests. Susceptibility to selected antimicrobial agents was detected by the disc diffusion method according to Clinical Laboratory Standards Institute recommendations. A total of 27 strains of Gram-negative rods were cultured from 21 patients. During short-term hospitalization, in patients with lung cancer undergoing thoracic surgery and preoperative prophylaxis, qualitative and quantitative changes in Gram-negative rods colonizing the nasopharynx were observed. A statistically significant increase in the frequency of these bacteria on mucous membranes of nasopharynx in examination II was found (Chi-squared test, p<0.00001). The strains of Enterobacteriaceae were highly susceptible to antimicrobial agents, whereas most of the non-fermenting rods were classified as multi-drug resistant organisms. PMID- 19123307 TI - Use of diode laser 980 nm as adjunctive therapy in the treatment of chronic periodontitis. A randomized controlled clinical trial. AB - The primary goal of periodontal therapy is the removal of supra and subgingival bacterial deposits by mechanical debridement consisting in scaling and root planing (SRP) using manual or power-driven instruments. The complete removal of bacteria and their toxins from periodontal pockets is not always achieved with conventional mechanical treatment. The use of lasers as an adjunctive therapy for periodontal disease may improve tissue healing by bactericidal and detoxification effects. The aim of this study was to compare the effectiveness of Diode laser used as adjunctive therapy of SRP to that of SRP alone for non surgical periodontal treatment in patients with chronic periodontitis. Nineteen pairs of teeth with untreated chronic periodontitis were selected in 13 patients and randomly treated by SRP alone (control group) or by SRP + laser irradiation (test group). Clinical measurements (PPD, CAL, BOP, GI, PI) were performed before treatment at baseline (T0) and at T1 (after 4 weeks), T2 (8 weeks), T3 (12 weeks), T4 (6 months). Subgingival plaque samples were taken at baseline and after treatment and examined for 8 periopathogens bacteria using PCR technique. The present study showed that the additional treatment with diode laser may lead to a slightly improvement of clinical parameters, whereas no significant differences between test and control group in reduction of periodontopathogens were found. PMID- 19123308 TI - Decrease of methicillin resistant Staphylococcus aureus prevalence after introduction of a surgical antibiotic prophylaxis protocol in an Italian hospital. AB - The use of antimicrobials is an important factor contributing to the emergence of antibiotic resistance. The goal of our study was to evaluate the impact of the introduction of an antibiotic surgical prophylaxis protocol on the prevalence of methicillin resistant Staphylococcus aureus (MRSA) infections in a tertiary care hospital. The protocol of surgical antibiotic prophylaxis was designed by a multidisciplinary team and was implemented in December 2001. Between January 2002 and December 2002, pharmacy, laboratory and active surveillance-records were prospectively reviewed to calculate prevalence rates of defined daily doses (DDD), microorganism isolation and health-care related infections. A progressive decrease from 1.58 to 0.56 of MRSA isolations per 1000 patient-days and from 76.4% to 29.4% MRSA prevalence rate was reported (p<0.001). Monthly prevalence rates of MRSA showed a significant linear correlation with the reduction of the DDD of the 3rd generation cephalosporins (r=0.90; p<0.001). MRSA surgical site and blood stream infections decreased from 78% to 38% and from 89% to 38%, respectively (p=0.017 and p=0.026). In our experience, the reduction of 3rd generation cephalosporin use was an effective strategy to reduce the MRSA infection rate and was associated with the reduction of the overall expenditure for antibiotics in the hospital. PMID- 19123309 TI - Prevalence of Coxiella burnetii antibodies in blood donors in Ankara, Central Anatolia, Turkey. AB - The aims of this study were to determine the prevalence of Coxiella burnetii antibodies among blood donors and to examine the epidemiological characteristics of C. bumetii infection in Ankara, Turkey. A total of 601 serum samples were collected from blood donors aged 18-61 years. Donor samples were stratified by age, sex, and residence (rural or urban). IgG and IgM antibodies to the C. bumetii phase II antigen were determined using a commercial ELISA. Blood samples reactive in the ELISA were also analysed using a commercial indirect immunofluorescence assay (IFA). The prevalence of anti-phase II IgG was 32.3%, and 17 (2.8%) were IgM positive. Seropositivity in men was higher than in women (33.2% vs. 21.7%, OR:1.88; 95% CI: 0.88-4.14) and the difference in seroprevalence rates between genders was not related to occupational exposure to domestic animals. 87.6% of seropositive donors reported no contact with farm animals. Our results revealed that C. burnetii infection is highly endemic in Ankara and that the majority of seropositive cases are not linked to specific occupational exposure in this area. In conclusion, the high rate of C. burnetii phase II antibodies among blood donors is a reflection of the high prevalence of Q fever in this area of Turkey and indicates the need for further studies, not only to determine the risk of transfusion-transmitted Q fever, but also to elucidate the epidemiology of Q fever in Turkey. These studies should be conducted through improved collaboration between the veterinary and medical services. PMID- 19123310 TI - In vitro activity of tigecycline as a therapeutic option against multidrug resistant Acinetobacter spp. AB - The study was performed to detect the in vitro activity of tigecycline in multidrug-resistant Acinetobacter isolates from patients in Hacettepe University Adult Hospital, Turkey. The microorganisms were isolated from clinical specimens of patients with respiratory and bloodstream infections. Thirty (66.7%) of the 45 inpatients were in ICUs. In vitro activity of imipenem, meropenem, ceftazidime, ciprofloxacin and aztreonam in 124 Acinetobacter species isolated was evaluated by microdilution test. Overall, 51 (41%) Acinetobacter spp. were found to be resistant to > or = 3 antibiotics belonging to different antimicrobial classes and defined as multidrug-resistant (MDR). Among the MDR Acinetobacter spp. 32 (62.7%) were Acinetobacter baumannii and 19 (37.3%) Acinetobacter lwoffii. In vitro activity of tigecycline against MDR isolates were studied by E-test. Each MDR isolate was also tested for metallo-beta-lactamase (MBL) production using CLSI guidelines. Forty-five (88.2%) of the isolates were found to produce MBL. The MIC90s of all antimicrobial agents tested except tigecycline were > or = 64 microg/ml whereas the MIC50, and MIC90 of tigecycline were found 1 microg/ml and 1.5 microg/ml, respectively. ERIC-PCR results revealed that bloodstream and respiratory isolates had nine and six different patterns, respectively. In conclusion, tigecycline has been shown to have potent in vitro activity against MDR Acinetobacter spp. and might be of therapeutic value in the treatment of infections due to MDR Acinetobacter spp., including those harbouring MBLs. Further clinical trials are needed to confirm the efficacy of tigecycline in the management of MDR Acinetobacter infections. PMID- 19123311 TI - Dermatophyte infections in patients attending a tertiary care hospital in northern Italy. AB - We studied dermatophyte infections in patients attending the Dermatology Outpatients Clinic of S. Matteo hospital Pavia, Italy, during the period 2004 2006. A total of 100 samples were collected from 95 patients; 97 dermatophytes and 3 keratinophylic fungi were isolated. Trichophyton rubrum was the most frequent dermatophyte isolated (42.3%), followed by Microsporum canis (31%), T. mentagrophytes (14.5%) and M. gypseum (9.2%). Less frequently isolated were Epidermophyton floccosum and T. violaceum. The most common dermatophyte infections included tinea corporis, tinea pedis, tinea unguium, tinea capitis and tinea cruris. Zoophilic dermatophytes were most commonly recovered from children and adolescents with tinea capitis and tinea corporis. Anthropophilic species were identified mostly in adults with tinea pedis, tinea cruris and onychomycosis. PMID- 19123312 TI - Serotyping of foot-and-mouth disease virus by antigen capture-ELISA using monoclonal antibodies and chicken IgY. AB - Laboratory detection of specific foot-and-mouth disease virus (FMDV) is routinely carried out by ELISA and RT-PCR. Identification and serotyping of FMDV by ELISA requires polyclonal antibodies raised in rabbits and guinea pigs. The polyclonal antibodies have certain disadvantages such as batch to batch variation, inconsistent yields of antibodies and limited quantity of serum obtained from individual animals. This paper describes a method wherein monoclonal antibodies and chicken IgY were used in an antigen capture-ELISA for serotyping of thirty tongue epithelial samples and sixty tissue culture fluids. The results were compared with the routine antigen detection ELISA. The present study indicated that monoclonal antibodies and chicken IgY can substitute conventional polyclonal antibodies for routine serotyping of FMDV. PMID- 19123313 TI - Phage type and antimicrobial susceptibility of Salmonella enterica serovar Enteritidis from food-producing animals in Japan between 1976 and 2004. AB - A total of 56 isolates of Salmonella enterica serovar Enteritidis, including 38 isolates from poultry, 16 from cattle and two from pigs, collected between 1976 and 2004, were subjected to bacteriophage typing and antimicrobial susceptibility testing. Phage type (PT) 8 was predominant in bovine isolates, whereas PT1 and PT4 were predominant in poultry isolates. Resistance was found for 8 of 11 antimicrobials tested, at the following rates: 46.4% for dihydrostreptomycin followed by ampicillin and oxytetracycline (both 8.9%). PMID- 19123314 TI - Laboratory detection of extended-spectrum beta-lactamase by an automated system. AB - This study aims to evaluate the positive predictive value (PPV) and the negative predictive value (NPV) of Vitek2 in detecting extended-spectrum beta-lactamase (ESBL) phenotypes when compared to a manual confirmatory test as gold standard. A sample of Escherichia coli, Klebsiella spp and Proteus mirabilis isolates were collected by 5 laboratories in the Emilia-Romagna Region (Italy). Vitek2 appears to be an accurate tool to detect ESBL phenotypes of E. coli isolates; some concern remains about its performance with the other bacterial species, especially P. mirabilis. PMID- 19123315 TI - Multiple malaria infection in a pregnant woman from Nigeria: detection by multiplex PCR. AB - For the last 100 years, diagnosis of malaria has been based on examination of Giemsa-stained thick and thin blood films under the microscope. This is a time consuming procedure which often fails to correctly diagnose the infecting species -especially when carried out by inexperienced technicians or when blood levels of parasite are low. Rapid Diagnostic Tests (RDTs) for antigen detection can distinguish between Plasmodium falciparum and Plasmodium vivax but cannot identify the species present in mixed infections. In the case reported here we used multiplex PCR to investigate suspected mixed infection in a pregnant woman from Nigeria. The results suggest that the method used is highly specific and can be very sensitive and that it has several advantages with respect to microscopy and RTDs. PMID- 19123316 TI - [Look on the research of the Chinese new drugs for treatment of chronic gastritis from "Shanghai Consensus"]. PMID- 19123317 TI - [Considerations on Chinese medicine research with reflections of "cyclopedia of Danshen"]. PMID- 19123318 TI - [Characteristics and treatment of older adults with acute myelocytic leukemia]. PMID- 19123319 TI - [Clinical features and treatment strategies of older patients with breast cancer]. PMID- 19123320 TI - [Research status of integrative internal medicine treatment of old patients with colorectal cancer]. PMID- 19123321 TI - [Analysis on associated risk factors and syndrome types in diabetes mellitus type 2 patients complicated with non-alcoholic fatty liver]. AB - OBJECTIVE: To explore the risk factors and the main TCM syndrome types associated with the diabetes mellitus type 2 (DM2) patients complicated with non-alcoholic fatty liver (FL). METHODS: Adopted controlled trial method, the age, stature, body weight, and body mass index (BMI) of 180 DM2 patients were compared with those complicated with or without FL. And some related laboratory indexes, including the age, stature, body weight, BMI, fasting blood glucose (FBG), C peptide (CP), glycosylated hemoglobin (HbA1c), triglyceride (TG), total cholesterol (TC), high and low density lipoprotein cholesterol (HDL-C and LDL-C), and 2 h post-prandial CP (2 h CP), were compared as well. Moreover, patients' TCM syndrome types were classified. RESULTS: No significant differences were found between DM2 patients complicated with or without FL in aspects of FBG, HbA1c, TC, LDL-C and age, stature (P > 0.05), but significant difference did show between them in aspects of CP (4.09 +/- 2.40 microg/L vs 2.47 +/- 1.74 microg/L), 2h CP (6.38 +/- 5.46 microg/L vs 4.35 +/- 2.92 microg/L), TG (2.81 +/- 2.33 mmol/L vs 1.93 +/- 1.92 mmol/L), HDL-C (1.07 +/- 0.06 mmol/L vs 1.19 +/- 0.32 mmol/L) as well as in body weight (73.4 +/- 11.7 kg vs 61.4 +/- 10.1 kg) and BMI (26.0 +/- 3.67 vs 22.8 +/- 3.23), respectively (P < 0.05 or P < 0.01). Moreover, phlegm dampness type was more liable to appear in DM2 patients complicated FL. CONCLUSIONS: Obesity, insulin resistance and lipid metabolism disorder are the chief risk factors in DM2 patients complicated with FL and phlegm-dampness is the chief pathogenesis. PMID- 19123322 TI - [Effect of Tangyikang in improving the function of pancreatic islet beta cells in patients with latent autoimmune diabetes mellitus in adults]. AB - OBJECTIVE: To investigate the effect and mechanism of Tangyikang (TYK) for improving pancreatic islet beta cell function in patients with latent autoimmune diabetes mellitus in adults (LADA). METHODS: Seventy-four LADA patients were randomly assigned to two groups. The 37 patients in the treatment group were treated with TYK decoction (one dose consisted of red ginseng 10 g, milkvetch root 30 g, lilyturf root 15 g, wild weed 10 g, coptis root 15 g, cape-jasmine fruit 10 g, giant knotweed rhizome 10 g, safflower 10 g and moutan bark 10 g) combined with insulin therapy, and the 37 in the control group treated with insulin therapy alone, and the course for all was 3 months. Changes of glycosylated hemoglobin, index of pancreatic islet beta-cell function (delta CP(2h)/delta BS(2h)), serum interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) were observed before and after treatment. RESULTS: All the above-mentioned indexes were improved after treatment in both groups, the post-treatment data showed significant difference between groups in delta CP(2h)/delta BS(2h), (0.258 +/- 0.106 vs 0.168 +/- 0.054, higher in the treatment group, t = 4.626, P < 0.01), but with insignificant difference in glycosylated hemoglobin (t = 0.441, P = 0.660). Besides, the dosage of insulin used in the treatment group was less than that in the control group (t = -4.169, P < 0.01); covariance analysis showed, through excluding impact of different dosages insulin used, IL- 4 level was higher (F = 24.217, P < 0.01) and IFN-gamma level was lower (F = 14.198, P < 0.01) in the treatment group than those in the control group. CONCLUSIONS: TYK could improve the function of islet beta-cell, its possible mechanism is related with the regulation on cell immunity and the correction of T-lymphocyte subsets (Th1/Th2 ratio) imbalance. PMID- 19123323 TI - [Visualized study on acupuncture treatment of children autism using single photon emission computed tomography]. AB - OBJECTIVE: To provide the molecular iconographic evidence for acupuncture treatment of children autism (CA). METHODS: Electric acupuncture (EA) was applied on acupoints of bilateral Hegu (LI4), Quchi (LI11), Zusanli (ST36) and Sanyinjiao (SP6) in 34 CA patients' with ABC scale > 57 scores. Using SIEMENS ECAM / ICON double head system, single photon emission computed tomography (SPECT) brain perfusion imaging were performed before and during EA with 99mTc-ECD. Then SPECT images were analyzed visually and quantitatively. RESULTS: Visual analysis showed that 114 foci of low cerebral perfusion and function were found in 34 CA patients before EA, 28 cases (82.35%) of them with abnormalities in left (or both) front lobes, left Broca area and left Wernicke area. During EA, the affected areas improved to different extent. Quantitative analysis showed that significant difference existed between pre-EA focal Ff and mirror-side Ff, pre-EA Ff and mid EA Fe, focal BFCR% and mirror-side BFCR % (all P < 0.01). The improving analysis of cerebral perfusion and function showed that the efficacy of EA was 78.95%. CONCLUSION: Acupuncture was proved preliminarily effective in treating children autism. PMID- 19123324 TI - [Effect of hallux-trampling combined with Yaotongling suppository on microcirculation in treating lumbar intervertebral disc prolapse]. AB - OBJECTIVE: To observe the effect of hallux-trampling (HT) combined with Yaotongling suppository (YS) on the microcirculation at waist region and lower limbs in treating patients with lumbar intervertebral disc prolapse (LIDP), and to preliminarily explore its mechanism. METHODS: Adopting randomized, double blinded medicine and single-blinded manipulation controlled method, 108 patients with LIDP of cold-accumulation and blood-stasis syndrome were assigned equally to 3 groups in random. Group A was treated with HT plus YS, Group B treated with HT plus placebo suppository, and Group C treated with YS plus mock HT. After 2-week treatment, the changes in score of symptoms and signs as well as the microcirculation at various site-points of waist and lower limbs were observed before and after treatment. RESULTS: The total scores of symptoms and signs after treatment in Group A, B and C was 7.340 +/- 4.627, 9.543 +/- 4.745 and 10.086 +/- 4.631 respectively, which was significantly lowered as compared with those before treatment respectively (P < 0.05). Significant difference of microcirculation between pre- and post-treatment was shown at all site-points in Group A and B (P < 0.05), but only shown at the site-points of lower limbs in Group C, so significant difference (P < 0.05) was shown when compared the microcirculation in Group A or B with that in Group C (P < 0.05). CONCLUSION: HT combined YS in treating LIDP could markedly improve the microcirculation at waist and lower limbs to correct the nutritional disturbance of nerve root, and the improvement of microcirculation is correlated with the changes of clinical symptoms and signs. PMID- 19123325 TI - [Effect of TCM therapy for invigorating Pi, soothing Gan, eliminating dampness and resolving blood stasis on the short-term quality of life in patients with diarrhea type irritable bowel syndrome]. AB - OBJECTIVE: To observe the TCM therapy for invigorating Pi, soothing Gan, eliminating dampness and resolving blood stasis (TCM-T) on the short-term quality of life (stQOL) in patients with diarrhea type irritable bowel syndrome (IBS-D). METHODS: One hundred and twenty-five patients with IBS-D were assigned to 3 groups, Group A was treated with TCM-T; Group B treated with some selected Chinese patent drugs according to patients' syndrome types, such as Changweikang granule, Guben Yichang tablet, Bupi Yichang pill and Shenling Baizhu pill, Group C treated with Western medicines as Dicetel 50 mg, Oryzanol 10 mg, and Bifid triple viable 420 mg, 3 times daily by oral administration. The therapeutic course was 4 weeks for all. The stQOL was evaluated and compared by SF-36 scale before and after treatment. RESULTS: Comparison between groups on the scores for the 8 dimensions of SF-36 scale, i.e., physiological function (PF), role physical (RP), body pain (BP), global health (GH), vitality (VT), society function (SF), emotional effect (RE), and mental health (MH), showed no significant difference before treatment, but scores of all the 8 dimensions were improved significantly in Group A after treatment (P < 0.05), with scores of SF, BP, GH and MH superior to those in Group B (P < 0.05, P < 0.01), and scores of RP, GH, SF, RE, PF and MH superior to those in Group C (P < 0.05, P < 0.01). CONCLUSION: TCM-T has favorable effect in treating IBS-D, it could improve the stQOL of patients. PMID- 19123326 TI - [Epidemiological survey on TCM syndrome distribution and season correlation of hypertension in Hunan Province]. AB - OBJECTIVE: To analyze the epidemiologic characteristics of TCM syndrome distribution and season correlation in hypertension patients. METHODS: Clinical materials regarding population, symptoms and signs, etc. of 698 hypertension patients were collected, and statistical analysis on them was conducted. RESULTS: TCM syndrome type revealed in patients, in order of frequency from high to low, were: Gan-fire flaming-up syndrome, yin-deficiency and yang-excess syndrome, phlegm-dampness accumulation syndrome, both yin-yang deficiency syndrome, and others. The incidence of various syndromes was higher in the winter/spring and rather low in the summer, and Gan-fire flaming-up syndrome was positively related to the spring and negative to the summer. CONCLUSIONS: The syndrome types of hypertension patients are chiefly the Gan-fire flaming-up syndrome, yin deficiency and yang-excess syndrome, phlegm-dampness accumulation syndrome, and both yin-yang deficiency syndrome; Gan-fire flaming-up syndrome is the dominance; their incidence is higher in the winter/spring and lower in the summer. PMID- 19123327 TI - [Regulatory effect of procyanidins on the expressions of interferon-gamma, interleukin-12 and transcription factor T-bet mRNA in peripheral blood mononuclear cell of patients with alopecia areata]. AB - OBJECTIVE: To investigate the regulatory effect of procyanidins (PC) on the expressions of Th1 type cytokines (IFN-gamma and IL-12) and the transcription factor T-bet mRNA in peripheral blood mononuclear cell (PBMC) in patients with alopecia areata (AA). METHODS: Above-mentioned expressions were detected using RT PCR technique in 20 AA patients (8 of mild and 12 of severe degree) and 10 healthy subjects after phytoaemagglutinin (PHA) or PHA + PC stimulation. RESULTS: After being stimulated by PHA and PHA + PC, in patients with severe AA, the expression of T-bet mRNA was 0.581 +/- 0.148 and 0.419 +/- 0.113 respectively; that of IFN-gamma mRNA, 0.689 +/- 0.219 and 0.430 +/- 0.162; and that of IL-12 mRNA, 0.198 +/- 0.056 and 0.136 +/- 0.035, respectively. As compared with those before stimulation, the respective difference was statistically significant (P < 0.05). CONCLUSIONS: PC can inhibit the expressions of Th1 type cytokines and transcription factor T-bet in PBMC of severe AA patients, and reverse the Th1 responses. The authors offered that it maybe part of the acting mechanism of pine needle for treatment of AA. PMID- 19123328 TI - [Effect of glycyrrhetinic acid on invasive capacity of leukemic cells and activity of gelatinase]. AB - OBJECTIVE: To study the effect of glycyrrhetinic acid (GA) on the invasive capacity of leukemic cells and the activity of intracellular gelatinase. METHODS: The effect of GA, in different concentrations, on the proliferation of cultured K562 and HL-60 leukemic cells in vitro was determined by MTT assay; that on cell invasive capacity was tested by Transwell cubicle matrigel invasion assay; and that on the activity of gelatinase in cells was detected by gelatin zymography. RESULTS: GA showed significant inhibitory effect on the proliferation of K562 and HL-60 leukemic cells; it inhibited the invasive capacity of cells in concentration-dependent manner; and significantly down-regulated the activity of gelatinase A and B in cells. CONCLUSIONS: GA can inhibit invasive capacity of K562 and HL-60 leukemia cells by way of suppressing the activity of gelatinase A and B. This study provides an experimental evidence for preventing extra medullary infiltration of leukemic cells. PMID- 19123329 TI - [Effects of matrine on invasion and metastasis of leukemia cell line Jurkat]. AB - OBJECTIVE: To study the inhibitory effects of matrine, in different concentrations, on invasion and metastasis of human acute lymphocytic leukemia cell line Jurkat. METHODS: In vitro cultured Jurkat cells were treated by matrine in concentration of 0 g/L, 0.1 g/L, 0.15 g/L and 0.2 g/L, respectively. Then cell adhesion assay, cell migration assay and matrigel invasion assay were used respectively to observe the effects of matrine on adhesion, migration and invasive capacity of Jurkat cells. Meantime, RT-PCR was performed to detect the matrix metalloproteinase (MMP)-2 and MMP-9 mRNA expression levels. Comparison of measurement data among groups was analyzed by variance analysis. RESULTS: As compared with the control group, the adhesion of Jurkat to fibronectin (FN) was significantly inhibited by 0.15 g/L and 0.2 g/L of matrine (P < 0.05); the cell migration and invasive capacity were significantly lowered by 0.1 g/L, 0.15 g/L and 0.2 g/L matrine (P < 0.01). High mRNA expression of MMP-9 presented but that of MMP-2 was expressed insignificantly in Jurkat cells, matrine at 0.1 g/L, 0.15 g/L and 0.2 g/L showed obvious effect in down-regulating MMP-9 mRNA expression (P < 0.01). Besides, MMP-9 mRNA expression was found to be positively correlated with the invasive capacity of Jurkat cells (r = 0.940, P < 0.01). CONCLUSIONS: Matrine is a good drug for antagonizing the invasion and metastasis of leukemia cells, it may roundly inhibit the adhesion, migration and invasive capacity of Jurkat cells, the mechanism might be related with the down-regulation of MMP-9 mRNA expression. PMID- 19123330 TI - [Effects of Xiongshao capsule on the proliferation of vascular smooth muscle cells in rabbits with atherosclerosis]. AB - OBJECTIVE: To study the effects of Xiongshao Capsule (XSC) on the proliferation of vascular smooth muscle cells (VSMC) in rabbits with experimental atherosclerosis (AS), and to explore its possible mechanisms. METHODS: Rabbit's fractional AS model was established by denuding and injuring the endodermis of abdominal aorta with 4F * Fogarty catheter, followed with feeding of high cholesterol forage. The animals were randomly divided into 8 groups, the model groups of 3 days, 2 weeks and 6 weeks after modeling (Group A, B and C); the single endothelium injury group (Group D), the probucol treated group (Group E), the low-dose and high-dose XSC treated groups (Group F and G) and the sham operative group (Group N). All were fed with high fat forage except those in Group N and D. The proliferative activity of neogenetic SMC at abdominal aorta with the most obvious pathological changes was analyzed by proliferating cell nuclear antigen immunohistochemical method; the VSMC phenotypic modulation was detected by transmission electron microscope (TEM), and the level of plasma angiotensin (Ang II) was measured by radioimmunoassay. And the effects of treatment on them were observed as well. RESULTS: The plasma Ang II level elevated gradually in Group A, and showed significant difference as compared that between Group C and Group N (P < 0.01); as compared with Group C, that in Group G was reduced significantly (P < 0.05); a reducing tendency was shown in Group E and F, but the difference of them with Group C was insignificant. Immunohistochemical dyeing showed that PCNA positive expressing cell was not found in the blood vessels of Group N, few was seen in Group A, while the upmost positive expression was shown in Group B, as for in Group C, significantly thickened endomembrane appeared, but the PCNA positive expression dropped. Number of PCNA positive cells reduced significantly (P < 0.05) in the drug treated groups, especially in Group G, showing significant difference as compared with that in Group C (P < 0.05, P < 0.01). TEM demonstrated normal shaped VSMC of aortic medial tunica in Group N, basically normal in Group A, but in Group B, migration of VSMCs into intima was found, as for in Group C, abundant lipid granules and bigger vacuoles appeared in VSMCs with markedly proliferated intercellular collagenous fibers. Synthetic transformation could also be found in Group D. The transform VSMCs in neogenetic endothelium was fewer in the drug treated groups than that in Group C. Morphology of VSMC was basically normal in Group G, with few intercellular collagenous fiber proliferation, while in Group E and F, many lipid droplets in VSMC and lot of collagenous fibers proliferation in intercellular space still retained. CONCLUSIONS: XSC can prevent the genesis and development of AS through significantly lowering the plasma Ang II level and inhibiting the migration and proliferation of VSMC. PMID- 19123331 TI - [Effect of Astragalus and Arctium in different combinations on reactive oxygen species content and nuclear transcription factor kappaB expression in renal tissue of streptozotocin rats]. AB - OBJECTIVE: To investigate the reactive oxygen species (ROS) content and nuclear transcription factor-kappa B expression (NF-kappaB) in renal tissue of diabetic rats and the effect of Astragalus and Arctium in combination on them. METHODS: Streptozotocin (STZ)-induced diabetic model rats were established. Astragalus and Arctium in different dosages (low, moderate, high) were combined after orthogonal design and administered respectively to the model rats via gastrogavage for 4 or 8 weeks. Content of ROS in the renal tissue was detected by flow cytometry and expression of functional NF-kappaB p65 was assessed by Western blot at the 4th and 8th weekends (4th week and 8th week). RESULTS: ROS content of renal tissue in the model rats was 36.55 +/- 7.46% at the 4th week and 31.91 +/- 5.83% at the 8th week, NF-kappaB p65 expression was 165.00 +/- 3.14 at the 4th week and 214.00 +/- 5.11 at the 8th week, all higher than those in normal rats (6.21 +/- 1.83% and 129.00 +/- 1.58 at the 4th week, 6.95 +/- 1.41% and 148.00 +/- 2.32 at the 8th week) respectively. The combined use of Astragalus and Arctium showed decreasing effects on both indexes significantly, and the decreasing effects of the combination with moderate and high dose Astragalus were better than those with low dose, with the details as follows: those of ROS at the 4th week were 11.43 +/ 2.42%, 18.37 +/- 7.58% and 22.10 +/- 4.71% for high, moderate and low dose Astragalus combination respectively (same hereinafter), at the 8th week 12.55 +/ 4.40%, 19.15 +/- 6.64% and 23.48 +/- 3.13%; and for NF-kappaB p65 expression at the 8th week, 185.00 +/- 6.99, 183.00 +/- 3.89 and 194.00 +/- 4.98 respectively. CONCLUSION: Combined use of Astragalus and Arctium may ameliorate the condition of diabetic nephropathy by inhibiting the activation of the ROS-NF-kappaB signal passage. PMID- 19123332 TI - [Effects of propyl gallate on serum inflammatory factors and protein expression of COX-2 and ICAM-1 in ischemic myocardium of rats with acute myocardial infarction]. AB - OBJECTIVE: To investigate the effects of Propyl Gallate (PrG) on serum inflammatory factors and protein expression of cyclooxygenase-2 (COX-2) and intercellular adhesion molecule-1 (ICAM-1) in ischemic myocardium of rats with acute myocardial infarction (AMI). METHODS: AMI model was induced by ligating the left anterior descending (LAD) branch of coronary artery in Wistar rats, and the perfect modeling was certified with ST segment elevation by standard limb lead II of electrocardiogram. Rats were randomly divided into 6 groups: Group A of normal rats, Group B of rats through sham operation, Group C of AMI model rats, Group D of model rats treated with high dose PrG (80 mg x kg(-1) x d(-1), via peritoneal injection), Group E of model rats treated with low dose PrG (40 mg x kg(-1) x d( 1), via peritoneal injection), and Group F of model rats treated with aspirin (25 mg x kg(-1) x d(-1), via gastrogavage), all the treatments were given in succession for 7 days. Radioimmunoassay (RIA) was used to determine serum contents of interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha), immunohistochemistry was used to determine the level of COX-2 and ICAM-1 protein expression in myocardium. RESULTS: Compared with Group B, the serum level of TNF alpha increased significantly, but not the level of IL-1beta in Group C. Besides, the COX-2 and ICAM-1 protein expressions in ischemic myocardium increased in Group C. All the above-mentioned changes were reversed to certain extent in Group E after treatment. CONCLUSIONS: PrG (40 mg x kg(-1) d(-1)) could decrease the serum level of inflammatory factor TNF-alpha, and slightly inhibit COX-2 and ICAM 1 protein expression in ischemic myocardium of AMI rats. PMID- 19123333 TI - [Comparative study on effects of combined massage-smouldering-washing therapy and mini-invasive surgery in treating knee osteoarthritis]. AB - OBJECTIVE: To comparatively study the curative effects of combined massage smouldering-washing therapy (MSW) and mini-invasive surgery in treating knee osteoarthritis (KOA) of mild-moderate degree so as to provide a suitable therapeutic protocol. METHODS: Sixty patients with KOA were assigned to two groups. The treatment group was treated with MSW once a day for 10 days as one course, and 4 courses were applied totally with an interval of 3 days between courses. The control group was treated with mini-invasive surgery by arthroscopic mopping, followed with post-operational intra-articular cavity injection with sodium hyaluronate injection, 20 mg every week for 5 times continuously. The therapeutic effect and the changes in scores of clinical symptoms and signs before and after treatment in the two groups were observed and compared. RESULTS: Outcome of 3-month follow-up showed the effective rate was 90% in the treatment group and 93.33% in the control group; scores of clinical symptoms and signs effectively improved in both groups, but the improvement on the 4 items (joint pain, swelling, soreness of loin and knee, and cold aversion of knee) was superior in the treatment group, while that on the other 4 items (pain during squatting or half-squatting, up stairs or down stairs, joint stiffiness and joint kinetic capacity) was superior in the control group (P < 0.05). CONCLUSION: Both MSW and mini-invasive surgery have definite curative effect on KOA but with different particularities. PMID- 19123334 TI - [Clinical analysis of 47 patients with psoriasis arthropathy treated by traditional Chinese medicine syndrome differentiation and integrative medicine]. AB - OBJECTIVE: To analyze the clinical characteristics of psoriasis arthropathy (PA) and to explore the effect of treatment by traditional Chinese medicine (TCM) syndrome differentiation and integrative medicine. METHODS: The initiating characteristics, clinical manifestations, related laboratory and X-ray data as well as the therapeutic efficacy of syndrome dependent TCM and integrative medicine on 47 PA patients were analyzed. RESULTS: Among all the 47 patients, 38 (80.6%) had the skin lesion revealed earlier to joint symptoms, the average time between the appearance of skin lesion and joint symptoms was 9.2 years; joint X ray abnormality was found in 32 cases (68.1%); and psoriasis nail involvement was found in 33 cases (70.2%). Seventeen patients were treated with TCM alone and obtained satisfactory results, while the other 30 patients, were treated by integrative medicine with the dosage of medicine reduced immediately after symptoms were relieved, and finally cured or improved. CONCLUSION: TCM and integrative medicine are effective in treating PA, with lesser side effects. PMID- 19123335 TI - [Colonic dripping with Taihuang liquid for treatment of neonatal hyperbilirubinemia]. AB - OBJECTIVE: To explore the clinical effects of colonic dripping with Taihuang liquid (THL) in treating neonatal hyperbilirubinemia (HBE). METHODS: One hundred and thirty-eight neonates with HBE were randomly assigned to two groups. Conventional treatment and nursing were given to both groups, and THL was given additionally to the observation group by colonic dripping. RESULTS: Significant differences between the observation group and the control group were shown in frequency of defecation (4.6 +/- 1.3 times/d vs 2.0 +/- 1.1 times/d), daily serum bilirubin reduction (31.5 +/- 10.1 micromol/L vs 23.3 +/- 8.3 micromol/L), and days for normalizing serum bilirubin level (5.6 +/- 3.5 d vs 7.8 +/- 4.1 d, all P < 0.01). CONCLUSION: Colonic dripping of THL could promote the excretion of bilirubin, so as to decrease the level of serum bilirubin in neonates with HBE. PMID- 19123336 TI - [Blood-stasis and toxin causing catastrophe hypothesis and acute cardiovascular events: proposal of the hypothesis and its clinical significance]. AB - A hypothesis of " blood-stasis and toxin causing catastrophe engender acute cardiovascular event (ACE)" was put forward according to TCM cognition on blood stasis and toxin, in combining with the up to date concept of atherosclerosis and coronary heart disease, and together with our clinical practical experiences. The etiology, pathogenesis, evolving law, initial characteristics, clinical manifestation, therapeutic methods, prescriptions and their compatibility, as well as the well-suited time for applying TCM intervention were discussed. The authors stressed that it is of great significance for further reducing the morbidity of ACE and improving the effect of integrative medicine for preventing and treating cardiovascular thrombotic disease. PMID- 19123337 TI - [Key points for clinical orientation in the secondary development of famous and superior Chinese patent drugs]. AB - Aimed at the ubiquitous problems in the secondary development of famous and superior Chinese patent drugs, the concept of clinical orientation is suggested by the authors. Its necessity in secondary development was explained and the method for establishment and confirmation was explored in this paper, which set up a favorable basis for secondary development of famous and superior Chinese patent drugs. PMID- 19123338 TI - [English translation of culture-specific lexicon commonly seen in Huangdi Neijing]. AB - HuangDi NeiJing, the Yellow Emperor's Classic of Internal Medicine, a foundational classic of Chinese medical theory, is ranked as the first among the four most important classics of traditional Chinese medicine. Translation of culture-specific Lexicon is a pivot, also a nodus in translating Huangdi Neijing, so, chewing over it is of important meaning for English translation of the Chinese classic or even for that of whole Chinese medicine. In expectation of somewhat contribution to the development of the Chinese medical English discipline, the classification and translation strategy for culture-specific lexicon were discussed by the authors in this paper through an empirical study on comparing the different terms translated in two versions of Huangdi Neijing translation. PMID- 19123339 TI - [Progress of researches on pharmacokinetics of active components of Chinese drugs]. AB - The pharmacokinetic research of Chinese drugs is still in the exploratory stage so far, a great progress has achieved in the researches on active components of them since the 1980s. The progresses in pharmacokinetic research of active components of some commonly used Chinese drugs were reviewed in this paper, and the problems to be solved in future were pointed out. PMID- 19123340 TI - [Effects of litter coverage and watering frequency on seed germination and seedling survival of Castanopsis fissa]. AB - Castanopsis fissa is an important pioneer species commonly used in the restoration of degraded subtropical grassland in southern China. The study on the effects of litter coverage and watering frequency on the seed germination and seedling survival of C. fissa showed that the effects of litter coverage depended on soil moisture condition. When the watering frequency was 1 time per day, litter covering could restrain seed germination and increase seedling mortality; while watering 2 or 3 times per day, the covering would ameliorate soil moisture condition, and benefit seed germination and seedling establishment. The biomass of C. fissa seedlings was enhanced significantly by litter covering. It was suggested that keeping a litter layer on soil surface could benefit the seed germination and seedling survival of C. fissa to restore subtropical shrub land. PMID- 19123341 TI - [Gap features and renewal dynamics in secondary natural Pinus tabulaeformis forest in hilly loess region]. AB - Taking the secondary natural Pinus tabulaeformis forest in hilly loess region as research object, the shape, size structure, distribution, gap-maker features, and renewal dynamics of gaps were investigated. The results showed that the areas of canopy gap (CG) and extended gap (EG) appeared to have a skewed and small gap- dominated distribution. The CGs had an average area of 31.15 m2, and those with an area of 20-40 m2 made up the highest proportions in number and area, accounting for 38.24% and 30.50%, respectively; while the EGs had an average area of 58.04 m2, and those with an area of 30-60 m2 made up the highest proportions in number and area, accounting for 36.77% and 27.79%, respectively. The average CG area accounted for 53.67% of the average EG area. The gaps were mainly elliptical, and their ages were mainly within 10-20 years, which occupied 33.82% of the total. The gaps were mainly with a height of 14-16 m, and those with a height of 18-22 m made up 36.8% of the total. The gaps were mainly formed by the trees being broken at their bases and the standing dead trees, which made up 47. 66% and 23.44% of all gap-makers, respectively, and thinning and unlawful felling were the major factors in gap formation. The tree deaths from lowered resistance due to tree- ageing, drought, and pest and diseases were one of the reasons for gap formation. The gap-makers per gap numbered 1.89 on average, and most of the gaps were formed by two gap-makers. The gap-makers were mainly Pinus tabulaeformis, followed by Populus davidiana, Betula platyphylla, and Quercus liaotungensis. The diameters of the gap-makers appeared to have a remarkably skewed normal distribution, and the diameters commonly ranged in 10-20 cm and 21 30 cm, taking up 25.0% and 45.31%, respectively. This skewed normal distribution agreed with the skewed area distribution of the gaps. In forest gap, trees had a better regeneration condition, and Chinese pine seedings had no age discontinuity; while in the understory of Chinese pine, seedings had an obvious discontinuity in age-structure. PMID- 19123342 TI - [Effects of exotic Larix kaempferi on forest soil quality and bacterial diversity]. AB - The study on the soil quality and bacterial diversity under 8-30 years old exotic Larix kaempferi, native Pinus tabulaeformis, and secondary deciduous broadleaf forest stands in Xiaolong-shan Mountains of Gansu, Northwest China showed that the soil pH under different forest stands had no distinct variation, but soil moisture content was increased with increasing age of forest stands. Soil organic matter and nitrogen contents were the highest under secondary deciduous forest, followed by under L. kaermpferi, and P. tabulaeformis. However, the soils under different ages of forest stands had no obvious variations in their organic matter and nitrogen contents, suggesting that tree species was the main factor affecting soil quality. Compared with P. tabulaeformis, exotic L. kaempferi could significantly increase soil organic matter and nitrogen contents. PCR-DGGE banding patterns suggested that the soil under secondary deciduous broadleaf forest had the highest bacterial diversity, followed by under L. kaempferi, and P. tabulaeformis. The sequenced DGGE bands were classified into three bacterial groups, i. e., Proteobacteria, Cytophaga - Flavobacterium - Bacteroides, and high G + C content gram-positive type, among which, Proteobacteria occurred most frequently. Further detailed analyses suggested that the soil bacterial compositions under exotic Larix stands were more similar to each other than those under pine and secondary deciduous broadleaf forests. It was concluded that exotic L. kaempferi induced the changes of microbial diversity in the forest soils of this region. PMID- 19123343 TI - [Characteristics of soil salinity profiles and their electromagnetic response under various vegetation types in coastal saline area]. AB - Aiming at the intrinsic relationships between vegetation type and soil salinity in coastal saline area, and by using electromagnetic induction EM38 and field sampling method, the characteristics of soil salinity profiles under various vegetation types in typical coastal saline region of the Yellow River Delta were analyzed, and the electromagnetic response characters of the salinity profiles were compared. The results showed that across the study area, soil salinity exhibited the characteristics of top enrichment and strong spatial variation. The horizontal electromagnetic conductivity EM(h) responded well to soil salinity at upper layers, and the response of vertical electromagnetic conductivity EM(v) to soil salinity at deeper layers was superior to that of EM(h). Soil salinity profiles were classified into inverted, normal, and uniform types. The vegetation types of inverted salinity profiles were mainly bare land and Suaeda salsa, while those of normal and uniform salinity profiles were cotton and weed, respectively. The sequence of top enrichment intensity was bare land > S. salsa land > weed land > cotton land. With the change of vegetation type of cotton-weed-S. salsa bare land, the EM(v)/EM(h) value of salinity profiles decreased gradually. Nonparametric test results showed that there was a significant correlation between vegetation type and electromagnetic response characters, and the distribution characters of EM(v)/EM(h) under various vegetation types varied significantly. PMID- 19123344 TI - [Seasonal variation of soil heat conduction in a larch plantation and its relations to environmental factors]. AB - Based on a 3-year (2003-2005) observation of soil heat flux (SHF) in a larch (Larix gmelinii) plantation, the characteristics of soil heat conduction in the plantation and their relationships with environment factors were analyzed. The results showed that there was an obvious seasonal variation of SHF in different years and sampling sites. The SHF was positive from April to August and mostly negative from September to next March, with an almost balance between heat income and outcome at annual scale. Solar net radiation had significant effects on the SHF and soil heat conductance (k), and an obvious time-lag effect was found, with 4-5 hours' time-lag in winter and 2-3 hours' time-lag in summer. Based on the real-time measurement of SHF and soil temperature difference at the study sites, the k value was significantly higher in early spring (P < 0.05), but no significant difference was observed in other seasons (P > 0.05). Therefore, when we use the observation data of soil temperature from weather stations to estimate soil heat flux, the k value in spring (from March to May) could induce a bias estimation. PMID- 19123345 TI - [Effects of different irrigation treatments on photosynthesis of Tieguanyin tea plants]. AB - By using chlorophyll fluorescence kinetics technique, the effects of different irrigation intervals, i.e., 5 d (T1), 10 d (T2), 15 d (T3), 20 d (T4), and 25 d (T5), on the photosynthesis of 2-year Tieguanyin tea plants were investigated in the field, with no irrigation as the control. The results showed that the leaf water potential and chlorophyll content decreased with increasing irrigation interval, while the net photosynthesis (P) increased first and decreased then, reaching its highest value (15.55, micromol x m(-2) x s(-1)) in treatment T2. The ratio of the variable to maximal fluorescence (F(v)/F(m)), the variable fluorescence quenching (deltaF(v)), and the variable fluorescence quenching rate (deltaF(v)/F(o)) all got the highest in treatment T2, being 0.844, 342.5, and 4.03, respectively. The initial fluorescence (F(o)) decreased with increasing irrigation interval, while a reverse changing trend of F(o) was observed in the control, demonstrating that PS II reaction center was damaged by drought stress. In conclusion, irrigating per 10 d was favorable to the photosynthetic electron transport and CO2 assimilation of the tea plants, which would enhance their photosynthesis efficiency. PMID- 19123346 TI - [Inhibition effects of salt stress on photosynthetic activity of Rumex K-1]. AB - With Rumex K-1 seedlings as test materials, this paper studied the effects of different concentration (100-300 mmol x L(-1)) NaCl and KCl on their leaf photosynthetic activity and osmotic adjustment. The results showed that at the concentration of 200 mmol x L(-1), NaCl had greater inhibition effect on the leaf photosynthetic activity than KCl, but at 300 mmol x L(-1), the inhibition effect of KCl was much greater than NaCl. After treated with 300 mmol x L(-1) of KCl and NaCl, the leaf water potential was -0.93 MPa and -1.05 MPa, and the osmotic potential was -1.43 MPa and -1.10 MPa, respectively, indicating that the increased damage caused by 300 mmol x L(-1) of KCl was not from osmotic stress. Under the stress of 300 mmol KCl x L(-1), the leaf Na+ concentration decreased by 88.6%, compared with the control, while the supplement of 25 mmol NaCl x L(-1) could obviously alleviate the damage of KCl on leaf photosynthesis, which proved that the deficit of Na+ and the accumulation of K+ in Rumex K-1 leaves could be responsible to the enhanced damage caused by 300 mmol x L(-1) of KCl. PMID- 19123347 TI - [Effects of NaCl on the growth and alkaloid content of Catharanthus roseus seedlings]. AB - Catharanthus roseus seedlings were grown in 1/2 Hoagland solution containing 0 250 mmol x L(-1) of NaCl, and their fresh and dry mass, malondialdehyde (MDA) and chlorophyll contents, tryptophan decarboxylase (TDC) and peroxidase (POD) activities, and vindoline, catharanthine, vincristine and vinblastine contents were measured after 7 days. The results showed that NaCl markedly decreased the fresh and dry mass but increased the MDA content. The chlorophyll content had no difference with the control when the concentration of NaCl was 50 mmol x L(-1), but decreased with increasing NaCl concentration when the NaCl concentration was above 50 mmol x L(-1). There was a significant enhancement of POD activity under NaCl stress. The TDC activity was the highest when the concentration of NaCl was 50 mmol x L(-1), but decreased with increasing NaCl concentration. The vindoline, catharanthine, vincristine, and vinblastine contents were the highest under 50 mmol x L(-1) NaCl stress, with the values being 4.61, 3.56, 1.19, and 2.95 mg x g(-1), respectively, and significant higher than the control and other treatments. Salt stress could restrain the growth of C. roseus seedlings, but promote the metabolism of alkaloid and increase the alkaloid content. 50 mmol x L(-1) of NaCl had the greatest promotion effect on the alkaloid content of C. roseus seedlings. PMID- 19123348 TI - [Effects of high temperature stress on photosynthesis and chlorophyll fluorescence of cut flower chrysanthemum (Dendranthema grandiflora 'Jinba')]. AB - Cut flower chrysanthemum (Dendranthema grandflora 'Jinba') plants were treated with 40 degrees C/35 degrees C or 33 degrees C/28 degrees C (day/night) for 11 days and then transferred to 23 degrees C/18 degrees C for 5 days to study the changes in their photosynthesis and fluorescence parameters under high temperature stress and normal temperature recovery. The results showed that on the 5th day of 33 degrees C/28 degrees C treatment, net photosynthesis (P(n)) decreased gradually and stomatal conductance (G(s)) decreased evidently; while after recovery for 5 days, both P(n) and G(s) resumed to 80% of the control. At 40 degrees C/35 degrees C, P(n) and G(s) decreased dramatically. The increase of intercellular CO2 concentration (C(i)) at the early stage under given high temperatures showed that the photosynthesis inhibition by high temperature stress was resulted from non-stomatal limitations. However, 9 days later, stomatal limitation became the mainly cause of photosynthesis inhibition. The intrinsic photochemical efficiency (F(v)/F(m)), quantum yield of PS II (phi(PS II), and the efficiency of excitation energy capture by open PS II reaction center (F(v)'/F(m)') at 33 degrees C/28 degrees C and 40 degrees C/35 degrees C all decreased, with antenna heat dissipation increased, indicating that reaction center was protected by decreased light capture and efficiency of electron transfer through PS II. The photochemical quenching (q(p)) at 33 degrees C/28 degrees C descended first and turned to rise then, suggesting that the electron transfer was firstly restrained by the stress. Contrastively, q(p) rose continuously at 40 degrees C/35 degrees C, indicating that oxygen-evolving complex (OEC) was the location in chrysanthemum photosynthesis apparatus most sensitive to extreme high temperature. PMID- 19123349 TI - [Effects of inoculation with arbuscular mycorrhizal fungi on AIPO4 uptake by Poncirus trifoliata]. AB - With sand culture in greenhouse, this paper studied the effects of inoculation with arbuscular mycorrhizal (AM) fungi Glomus mosseae and G. versiforme on the utilization of insoluble phosphate AlPO4 (Al-P) by trifoliate orange (Poncirus trifoliata) seedlings. The results indicated that inoculation with AM fungi notably increased the dry matter, P content, and P absorption rate of P. trifoliata. The contribution of mycorrhiza, P uptake by plant and AM fungi, and the contribution of AM fungi in P uptake by P. trifoliata increased greatly with increasing Al-P level in the medium. Inoculation with AM fungi greatly increased the excretion of phosphatase, especially acid- and neutral phosphatase, from plant root and hyphae, but the excretion amount decreased with increasing Al-P level. The contribution of the fungi in P uptake by P. trifoliata was significantly positively correlated with the amounts of acid-, neutral-, alkaline , and total phosphatase. PMID- 19123350 TI - [Simulation on the seasonal growth patterns of grassland plant communities in northern China]. AB - Soil moisture is the key factor limiting the productivity of grassland in northern China ranging from arid to subhumid arid regions. In this paper, the seasonal and annual growth, foliage projective cover (FPC), evaporative coefficient (k), and net primary productivity (NPP) of 7 types of grasslands in North China were simulated by using a simple model based on well established ecological processes of water balance and climatic data collected at 460 sites over 40 years. The observed NPPs were used to validate the model, and the simulated NPPs were in high agreement with the observed NPPs. The simulated k, NPP, and FPC deceased from east to west in temperate grasslands, and decreased from southeast to northwest in Qinghai-Tibet Plateau, reflecting the moisture gradient in northern China. Alpine meadow had the highest k, NPP, and FPC in the 7 types of grasslands, alpine steppe had the second highest FPC but with a NPP similar to that of temperate steppe, and the three simulated parameters of temperate desert were the smallest. The simulated results suggested that the livestock density should be lower than 5.2, 2.3, 3.6, 2.1, 1.0, 0.6, and 0.2 sheep unit x hm(-2), while the coverage of rehabilitated vegetation should be about 93%, 79%, 56%, 50%, 44%, 38%, and 37% in alpine meadow, alpine steppe, temperate meadow steppe, temperate steppe, temperate desert steppe, temperate steppe desert, and temperate desert, respectively. PMID- 19123351 TI - [Net primary productivity of Leymus chinensis steppe in Xilin River basin of Inner Mongolia and its responses to global climate change]. AB - CENTURY model was utilized to simulate the annual aboveground net primary production (ANPP) of Leymus chinensis steppe, a dominant community type in Xilin River basin of Inner Mongolia steppe region. The results showed that the model performed reasonably well in predicting the dynamics of the ANPP. The scenario based simulations indicated that though the variations of air temperature and precipitation due to global climate change as well as the elevated CO2 would significantly affect the dynamics of the ANPP, precipitation was the key affecting factor. Several GCM models had predicted that the precipitation in this region would decrease in the future, and consequently, it was likely that the ANPP would also decrease. Nevertheless, the simulation results showed that while the ANPP decreased in most climate change scenarios, it might also increase in the following climate change scenarios: 1) if the atmospheric CO2 concentration was doubled, air temperature was increased by 2 degrees C, and precipitation was kept unchanged or increased by 10%-20%, and 2) if the atmospheric CO2 concentration was kept unchanged, air temperature was increased by 2 degrees C, and precipitation was increased by 20%. Overall, it was evident that climate change would have significant effects on the steppe in Xilin River basin of Inner Mongolia. PMID- 19123352 TI - [Micro-community characteristics of vegetations in blowouts and depositional areas of Hulunbuir grassland, Inner Mongolia]. AB - By using traditional sampling methods, the micro-communities of vegetations in fixed, semi-bare, and bare blowouts of Hulunbuir grassland were investigated, and the investigation data were statistical analyzed. The results showed that the vegetation coverage decreased in the order of fixed blowout, semi-bare blowout, and bare blowout, and was lower than that of the primary vegetation Form. Stipa grandis. Potentilla acaulis and Kengia squarrosa were the dominant species in fixed blowout, with the coverage being 5%; while P. acaulis and Carex sp. were the dominant species in semi-bare blowout, with the coverage being 2%. The dominant species in depositional areas of semi-bare blowout were P. acaulis, K. squarrosa, Agropyron cristatum, and Thymus mongolicus, and the coverage was 4%. The dominant species on the southwest slope of bare blowout was Agriophyllum pungens. The middle depositional area of bare blowout was also occupied by A. pungens (coverage 4.7%), and the edge of it was dominated by A. cristatum (coverage 2.7%), Carex sp. (coverage 2.6%), and T. mongolicus (coverage 1.7%) from the edge of the depositional area to primary grassland. The mean species importance value in fixed, semi-bare, and bare blowouts was 12.64%, 13.38%, and 20.08%, while that in the depositional area of semi-bare blowout and in the middle and edge of bare blowout was 12.55%, 40.18%, and 11.15%, respectively. PMID- 19123353 TI - [Assessment on vegetation restoration capacity of several grassland ecosystems under destroyed disturbance in permafrost regions of Qinghai-Tibet Plateau]. AB - Vegetation restoration capacity of disturbed grassland ecosystem is one of the important components in assessing the influence of human engineering activities on the grassland ecosystems in permafrost region of Qinghai-Tibet Plateau. After comparing the features of vegetation communities, plant species diversity, grassland primary productivity, and economic group between disturbed and undisturbed communities, the comprehensive vegetation restoration capacity of several grassland communities under destroyed disturbance was assessed by using comprehensive assessment index. The results showed that the restoration of cover and community composition was obvious after 26 years natural restoration, being better for alpine steppes than for alpine meadows. However, the cover of disturbed communities was less than that of undisturbed communities. The restoration of plant species diversity in Stipa purpurea steppe was better than that in other grassland types. The biomass of standing plants restored from 0 to 148.8-489.6 g x m(-2), and that of Kobresia tibetica meadow was the highest (489.6 g x m(-2)). The palatable plants of disturbed communities were lower than those of undisturbed communities except for K. tibetica meadow, in which, the palatable plants were not changeable between disturbed and undisturbed communities. The comprehensive vegetation restoration capacity of alpine steppes was better than that of alpine meadows. PMID- 19123354 TI - [Diurnal dynamics of photosynthetic characteristics of alfalfa on Kerqin sandy land]. AB - By using a Li-6400 portable photosynthesis system, the diurnal dynamics of the photosynthetic characteristics of three alfalfa varieties Medicago sativa L. cv. Aohan, M. sativa L. cv. Rangelander, and M. sativa L. cv. Algonquin were studied. The results showed that for the test alfalfa varieties, the diurnal variation curves of their P(n), T(r), and WUE were not all presented double-peak. For Aohan and Algonquin, the maximum P(n) and T(r) occurred at 9:00-11:00; while for Rangelander, they appeared at 11:00. The minimum P(n) and T(r) of the three varieties all appeared at 7:00 and 19:00. The maximum WUE of Rangelander and Algonquin appeared at 7:00 and that of Aohan was at 9:00, and the minimum WUE of the three varieties all appeared at 15:00-17:00. Comprehensive analysis on the photosynthetic performance indices P(n), T(r), and WUE indicated that 2 years old alfalfa was in the best state, followed by 1 year old, and 4 years old. Correlation analysis showed that air temperature (T(a)) was the main environmental factor affecting P(n) and T(r), followed by water vapor pressure deficit (VPD) and relative humidity (RH). PMID- 19123355 TI - [Growth responses of belowground modules of Carex lasiocarpa to different water regimes and water experiences]. AB - With seedling's transplanting experiment under different water levels, this paper studied the growth responses of belowground modules of Carex lasiocarpa to various water regimes and water experiences in Sanjiang Plain. The results showed that the belowground modules of C. lasiocarpa had significantly different responses to water regimes. At thriving stage, the length of rhizome and adventitious root decreased with increasing water level, and until later growth stage, the maximal value still appeared under drought condition. However, under dry-wet alternate condition, the length of rhizome and adventitious root increased most from thriving stage to the end, indicating that stable and lower water level could improve the growth of rhizome and adventitious root. The biomass of rhizome, adventitious root, and belowground part were maximal under dry-wet alternate condition at both growth stages. For those with different water experiences, the ones undergoing alternate condition in early growth season and then drought had maximal rhizome biomass, and the others under sustained alternate condition had maximal adventitious root and belowground biomass. More biomass was distributed to rhizome in the later growth season under various water regimes. The percentage of rhizome in total biomass was significantly higher under drought condition than under other water conditions through the growth season. Besides, C. lasiocarpa grew slowly when submerged, but could recover through rhizomatic reproduction after the stress disappeared. PMID- 19123356 TI - [Accuracy of winter wheat identification based on multi-temporal CBERS-02 images]. AB - Chinese-Brazil Earth Resources Satellite No. 2 (CBERS-02) has good spatial resolution and abundant spectral information, and a strong ability in detecting vegetation. Based on five CBERS-02 images in winter wheat growth season, the spectral distance between winter wheat and other ground targets was calculated, and then, winter wheat was classified from each individual image or their combinations by using supervised classification. The train and validation samples were derived from high resolution Aerial Images and SPOT5 images. The accuracies and analyses were evaluated for CBERS-02 images at early growth stages, and the results were compared to those of TM images acquired in the same phenological calendars. The results showed that temporal information was the main factor affecting the classification accuracy in winter wheat, but the characteristics of different sensors could affect the classification accuracy. The multi-temporal images could improve the classification accuracy, compared with the results derived from signal stage, with the producer accuracy of optimum periods combination improved 20.0% and user accuracy improved 7.83%. Compared with TM sensor, the classification accuracy from CBERS-02 was a little lower. PMID- 19123357 TI - [Spatiotemporal variation of root systems of cytoplasmic male sterile maize (Zea may L.) and its homotype fertile lines]. AB - Maize cytoplasmic male sterile (CMS) lines and their homotype fertile lines were cultivated on soil column to study the differences of their root traits at different growth stages after anthesis. The results indicated that CMS lines had stronger ability of dry matter accumulation in their aboveground part, their grain yield was significantly higher than that of homotype fertile lines (P < 0.05), and their root dry mass was higher. In addition, the ratio of their root to canopy had a similar variation trend as root dry mass. The root mass of CMS lines in 0-20 cm and 20-40 cm soil layers was higher than that of homotype fertile lines, and the superiority was more obvious in 40-80 cm soil layer. Compared with homotype fertile lines, CMS lines had significantly higher root activity in 0-20 cm and 20-40 cm soil layers (P < 0.05), and significantly higher root SOD activity and soluble protein content (P < 0.05) in 40-80 cm soil layer. The root activity and root SOD activity and soluble protein content of CMS lines were significantly higher than those of their homotype fertile lines (P < 0.05) at late growth stage. In conclusion, CMS lines had the characteristics of larger root quantity in deeper soil layers and higher root physiological activity at late growth stage, which could prolong the functional period of root and postpone the process of senescence, possibly being one of the important reasons of the high grain yield of CMS lines. PMID- 19123358 TI - [Effects of nitrogen application rate on spatiotemporal variability of biomass accumulation of cotton's fruiting branch at flower and boll stage]. AB - Field experiments were conducted in Anyang and Nanjing, the typical representatives of ecological regions in the Yellow River Valley and the lower reaches of Yangtze River Valley in China, to quantitatively analyze the spatiotemporal variability of biomass accumulation of the fruiting branch of cotton variety Nucotn 33B at its flower and boll stage under effects of 0, 120, 240, 360, and 480 kg N x hm(-2) (marked as N0, N1, N2, N3, and N4, respectively). The results indicated that both in Nanjing and in Anyang, the temporal variability of biomass accumulation of vegetative and reproductive organs at different positions of fruiting branch could be described by Logistic equation, but the spatial variability was obviously different. Under the nitrogen application rate of 360 kg x hm(-2) in Anyang and of 240 kg x hm(-2) in Nanjing, the biomass accumulation had the characteristics of earlier starting date, shorter duration, and maximal speed of rapid increase. These nitrogen application rates could accelerate the biomass accumulation to benefit the high lint yield and fiber quality, while excess or deficient nitrogen supply was disadvantageous to the biomass accumulation of fruiting branch. Therefore, it could be possible to apply appropriate amount of nitrogen to regulate the growth eigenvalues of different positions fruiting branch at its speed growth stage to obtain high lint yield and fiber quality. PMID- 19123359 TI - [Chemical forms and subcellular and molecular distribution of Cd in two Cd accumulation rice genotypes]. AB - A hydroponic experiment combined with subcellular fractionation and gel filtration was carried out to study the chemical forms and subcellular and molecular distribution of Cd in rice roots and leaves, and to compare the differences between the rice varieties 'Guangyuanzhan No. 3' (a low-Cd accumulation genotype) and 'Zhenguiai' (a high-Cd accumulation genotype). The results showed that with increasing concentration of Cd in the medium, the Cd content in all fractions of rice roots and leaves increased significantly, and most of Cd accumulated in F I (cell wall) and F III (soluble substance). Compared with 'Guangyuanzhan No. 3', 'Zhenguiai' had a significantly higher soluble fraction of Cd in its roots and leaves. The greatest amount of Cd in roots and leaves was in the extraction solution of 1 mol x L(-1) NaCl, followed by in 2% HAC, and 0.6 mol x L(-1) HCl. In comparison with 'Guangyuanzhan No. 3', 'Zhenguiai' had a higher proportion of d-H2O and 80% ethanol extractable-Cd, the more mobile form of Cd. In the two rice varieties, the soluble Cd eluted followed the same pattern as the soluble protein in the sample medium, and the majority of soluble Cd was bounded to phytochelatins (PCs) or low molecular weight components. However, the Cd-PCs content in 'Guangyuanzhan No. 3' roots was much lower than that in 'Zhenguiai' roots, which could possibly result in a lower Cd mobility in roots and less Cd translocation from root to aboveground part. PMID- 19123360 TI - [Effects of simulated acid rain on leaf photosynthate, growth, and yield of wheat]. AB - With winter wheat variety Yamgmai 12 as test object, a field experiment was conducted to study the stress of simulated acid rain on its growth and development. The results showed that simulated acid rain had considerable effect on wheat growth and yield. When the pH of acid rain was < or = 3.5, the growth of leaf area as well as the mass of fresh leaf per unit area declined greatly, and the yield was significantly lower than CK. When pH was < or = 2.5, the plant height was obviously lowered, and the visible injury on leaf surface was observed. Under acid rain stress, the contents of leaf chlorophyll a, chlorophyll b, and carotenoid, especially chlorophyll a, decreased obviously. Acid rain also suppressed the synthesis of soluble sugar and reduced sugar, and the suppression was stronger at pH < or = 3.5, and became much stronger with increasing acidity. The total free amino acid and soluble protein contents in leaves decreased with increasing acidity, and were significantly lower than CK when the pH was < or = 3.5 and < or = 4.5, respectively. PMID- 19123361 TI - [Changes of microbial biomass carbon and enzyme activities in rice-barnyard grass coexisted soils]. AB - Taking barnyard grass (Echinochloa crus-galli), allelopathic rice (Oryza sativa) PI312777, and non-allelopathic rice Liaojing-9 as test materials, this paper studied the changes of microbial biomass C and of dehydrogenase, urease and invertase activities in soils under 1 : 1 rice-barnyard grass coexistence. The results showed that with the coexistence of barnyard grass, the microbial biomass C in rhizosphere soil of PI312777 was decreased by 50.52% (P < 0.01), while that in inter-row soil was increased. The microbial biomass C in rhizosphere soil of Liaojing-9 was also decreased by 38.99% (P < 0.01), but no significant difference was observed in inter-row soil. The dehydrogenase activity in rhizosphere soils of the two rice varieties were decreased by 20% or more (P < 0.05), but the urease and invertase activities in rhizosphere soil of PI312777 were significantly stimulated (P < 0.01). The invertase activity in rhizosphere soil of Liaojing-9 was inhibited by coexisting barnyard grass, but urease activity had less change. As a result, the allelopathic traits of rice variety led to a significant decrease of microbial biomass C and an increase of urease and invertase activities in rhizosphere soil under barnyard grass coexistence, implying that soil microbial biomass and enzymes might play essential roles in species interactions between rice and barnyard grass, and allelopathic rice had its advantage in resistance to barnyard grass. PMID- 19123362 TI - [Assessment on the availability of nitrogen fertilization in improving carbon sequestration potential of China's cropland soil]. AB - With reference to the situation of nitrogen fertilization in 2003 and the recommendations from agricultural experts on fertilization to different crops, two scenarios, namely, 'current situation' and 'fertilization as recommended', were set for estimating the current and potential carbon sequestration of China's cropland soil under nitrogen fertilization. After collecting and analyzing the typical data from the long-term agricultural experiment stations all over China, and based on the recent studies of soil organic matter and nutrient dynamics, we plotted China into four agricultural regions, and estimated the carbon sequestration rate and potential of cropland soil under the two scenarios in each province of China. Meanwhile, with the data concerning fossil fuel consumption for fertilizer production and nitrogen fertilization, the greenhouse gas leakage caused by nitrogen fertilizer production and application was estimated with the help of the parameters given by domestic studies and IPCC. We further proposed that the available carbon sequestration potential of cropland soil could be taken as the criterion of the validity and availability of carbon sequestration measures. The results showed that the application of synthetic nitrogen fertilizer could bring about a carbon sequestration potential of 21.9 Tg C x a( 1) in current situation, and 30.2 Tg C x a(-1) with fertilization as recommended. However, under the two scenarios, the greenhouse gas leakage caused by fertilizer production and application would reach 72.9 Tg C x a(-1) and 91.4 Tg C x a(-1), and thus, the actual available carbon sequestration potential would be -51.0 Tg C x a(-1) and -61.1 Tg C x a(-1), respectively. The situation was even worse under the 'fertilization as recommended' scenario, because the increase in the amount of nitrogen fertilization would lead to 10. 1 Tg C x a(-1) or more net greenhouse gas emission. All these results indicated that the application of synthetic nitrogen fertilizer could not be taken as a feasible measure for the carbon sequestration of cropland soil in China. Since synthetic fertilizer application is the basic guarantee of China's crop production, it was suggested to increase the efficiency of synthetic nitrogen fertilizer, and at the same time, to cut down the synthetic nitrogen fertilizer production and its application on the premise that the crop yield should be ensured. PMID- 19123363 TI - [Dynamic changes of ecosystem service value in Pinggu District of Beijing]. AB - Based on the land use/cover data of 1995 and 2004, and by using ecosystem service value per unit area of different terrestrial ecosystem types in China, the dynamic changes of ecosystem service value in Pinggu District of Beijing were analyzed. The results showed that the total ecosystem service value of Pinggu District was decreased from 3.291 x 10(9) yuan in 1995 to 3.044 x 10(9) yuan in 2004, with a change rate of -7.50%. The decrease in the areas of farmland and waters was the primary cause of the loss in ecosystem service function. Aquatic ecosystem had the highest ecological service value per unit area, while forest ecosystem gave the greatest contribution to the total ecosystem service value. The dynamic changes of the ecological service value revealed the conspicuously irrationality of land use structure in the District. The area proportions of forestland, farmland, and waters should be increased, and the conversion from farmland to constructive land should be controlled strictly. As the emergency water source and eco-conservation division of Beijing City, Pinggu District should improve its eco-environment protection and economic development to support the ecological and drinking water source securities of the Capital Beijing. PMID- 19123364 TI - [Emergy analysis of agro-ecolomic system in Shanxi Province]. AB - By the methods of emergy, this paper studied the input and output, working efficiency, and environmental loading of the agro-ecolomic system in Shanxi Province in 2005. The results showed that in 2005, the agro-ecolomic system in Shanxi still stayed in the period of traditional agriculture, which mainly depended on manpower and environmental resources. The emergy investment ratio (EIR) was 1.07, emergy yield ratio (EYR) was 0.99, and environmental loading ratio (ELR) was 6.55. In the structural adjustment of agriculture, stockbreeding had made great strides forward, but grain crops other than rice and wheat, vegetables, and fruits still had smaller emergy yield and were far from becoming dominant industry. The efficiency of the agro-ecolomic system was lower, and its environmental loading press was bigger. For the future, the surplus labors in agriculture in Shanxi should be shifted continually to other industries, and the high quality emergy such as agricultural science and technology should be increased to improve the use efficiency of environmental resources and the input and output of the system emergy. PMID- 19123365 TI - [Effects of leachate-loading and irrigation depth on N2O and CO2 emissions from leachate-irrigated soil]. AB - By using prefabricated probe and static chamber, the N2O concentration at different depths along the soil columns irrigated by landfill leachate as well as the N2O and CO2 fluxes were determined. The results showed that leachate irrigation could promote the N2O emission. Within the first 24 hours after irrigation, the N2O fluxes from the soil columns were significantly correlated with the N2O concentrations in the columns at the depths of 10 cm (r = 0.944, P < 0.01), 20 cm (r = 0.799, P < 0.01), 30 cm (r = 0.666, P < 0.01), and 40 cm (r = 0.482, P < 0.05), and the correlation decreased along the depths. Landfill leachate irrigation also promoted the CO2 emission from the soil columns. There was no significant correlation between the emissions of N2O and CO2 (P > 0.05). The irrigation loading of landfill leachate had a predominant effect on the total greenhouse gas emissions (including N2O and CO2 fluxes, based on CO2-equivalent emissions). When the irrigation loading was 6 mm x d(-1), the total greenhouse gas emissions from the soil columns were 2 times higher than those when the irrigation loading was 2 mm x d(-1). 47% of the total greenhouse gas emissions were reduced from the soil columns when the leachate irrigation depth was at 20 cm, compared with the depth being at 10 cm. Within the 14 days under leachate irrigation, 57.0%-91.0% of the total greenhouse gas emissions were originated from N2O emission. PMID- 19123366 TI - [Application of frost number model in Northeast China pepmafrost regionalization]. AB - Based on the 1981-2000 observation data of climate factors and snow depth in Northeast China, frost number model was applied to approach its applicability in the regionalization of the permafrost in this area. The results indicated that the permafrost in Northeast China was dominated by discontinuous permafrost, including large piece permafrost, large piece and island-like permafrost, and alpine permafrost. The comparison of permafrost regionalization maps in different periods showed that the simulation results of the model could reflect the recent distribution status of the permafrost in Northeast China. The southern boundary of the discontinuous permafrost could reach to Aershan Mountain region, and the other alpine permafrost as well as the permafrost of Yichun in Xiaoxing'an Mountains could be also distinguished by the model. PMID- 19123367 TI - [Simulation model on the formation of greenhouse sweet pepper leaf area index]. AB - Leaf area index (LAI) is one of the most important crop parameters in photosynthesis-driving crop growth simulation model and canopy evapotranspiration simulation model, while air temperature and radiation are the important climate factors affecting crop leaf growth. In this paper, experiments with different sweet pepper (Capsicum annuum L.) cultivars and sowing dates were conducted in greenhouse to quantitatively analyze the relationships of the number of unfolding leaves per plant, the number of old leaves removed per plant, and the length of each leaf with air temperature and radiation. Based on these quantitative relationships, a leaf area simulation model for greenhouse sweet pepper was developed, and the independent experimental data were used to validate the model. The results showed that the number of unfolding leaves per plant was a positive exponential function of the product of thermal effectiveness and PAR (TEP) accumulated after emergence, and the length of each leaf was a negative exponential function of the TEP accumulated after emergence. The coefficient of determination (R2) and the root mean squared error (RMSE) between simulated and measured leaf number, leaf length, and LAI were 0.94, 0.89, and 0.93, and 3.4, 2.15, and 0.15, respectively. The model could use air temperature, radiation, planting density, and emergence date to satisfactorily predict the LAI of greenhouse sweet pepper, and supply required LAI information for the sweet pepper growth and canopy evapotranspiration simulation models. PMID- 19123368 TI - [Characteristics of ichthyoplankton assemblages in Yangtze Estuary and adjacent waters in spring]. AB - Based on the investigation data of ichthyoplankton assemblages and environmental factors in Yangtze Estuary and adjacent waters in May 1999 and 2001, the characteristics of ichthyoplankton assemblages in these areas in spring were studied by using TWINSPAN (two-way indicator species analysis) and CCA (canonical correspondence analysis). A total of 11 540 ichthyoplankton individuals were taxonomically identified, belonging to 11 orders, 18 families and 32 species, of which, Coilia mystus, Engraulis japonicus, Chaeturichthys hexanema, Allanetta bleekeri, and Trachidermis fasciatus were the dominant species. The ichthyoplankton communities were classified into three assemblages by using TWINSPAN, i.e., estuarine assemblage dominated by C. mystus, coastal assemblage dominated by A. bleekeri and T. fasciatus; and shelf assemblage featured by E. japonicus and C. hexanema. The CCA ordination of the interrelations among the three assemblages and their correlations to the environmental variables revealed that salinity, depth, dissolved oxygen, and total suspended particulate matter were the major factors affecting the ichthyoplankton assemblages in the study areas. PMID- 19123369 TI - [Simulation of algae bloom under different flow velocity]. AB - The construction of a river-type reservoir will normally cause a change of hydrodynamic condition and a decrease of flow velocity in branch bays, leading to the formation of algae bloom when appropriate environmental conditions and sufficient nutrient supply are encountered, and thereby, causing the deterioration of fluvial ecosystem therein. To investigate the effects of hydrodynamic conditions on algae bloom, flow velocity was taken as the characteristic parameter, and controlled differently in the closed and recurrent models under identical environmental and nutrient conditions. The results showed that the increase of the flow velocity in a range of < 0. 4 m x s(-1) would accelerate the growth of algae and the occurrence of bloom. Under the conditions of different flow velocity, the growth of algae was similar to the common process of other microorganisms, namely, lag phase firstly, exponential phase secondly, stationary phase thirdly, and decline phase lastly. Accordingly, the outbreak peak and regression floor of the algae bloom would occur. Therefore, the increase of flow velocity in a range of < 0. 4 m x s(-1) would not inhibit the growth of algae and the occurrence of bloom. The effective characteristic parameter of the hydrodynamic conditions would be studied in depth in future. PMID- 19123370 TI - [Distribution characteristics of benthic algae in intertidal zone of Ma' an Archipelago of Zhejiang Province]. AB - Based on the survey of benthic algae in the intertidal zone of Ma' an Archipelago from March to July 2007, the algal species composition, distribution, and temperature feature were studied. The dominant algal species in the study area were preliminarily analyzed by using similarity indices (S(c)) and index of relative importance (IRI(c)). A total of 31 species sampled in sublittoral area were identified, among which, 7 species of 5 genera belonged to Chlorophyta, 8 species of 5 genera belonged to Phaeophyta, and 16 species of 14 genera belonged to Rhodophyta. Topical and selective distribution species influenced by wave and tide were identified in the intertidal zone. Ulva pertusa and Sargassum thunbergii were found in all survey area. Rhodophyta was the dominant species, with the occurring frequency being up to 61.1%, and Chlorophyta showed quite uniformed horizontal distribution. In addition, 81% of sampled species were from low-tide zone, and some were extended from mid-tide zone to low-tide zone. The composition comparability between mid-tide and low-tide species was 0.47, and the convergence effect in mid-tide and low-tide zone was higher than that in high tide and mid-tide zone. The sublittoral area of Ma' an Archipelago showed obvious vertical zoning character, with temperate species being absolute abundant, and the warm-water species dominant. The marine floral texture of Ma' an Archipelago belongs to warm temperate-subtropical transitional marine flora. PMID- 19123371 TI - [Silviculture of poplar plantation in China: a review]. AB - There are more than 7.0 million hm2 of poplar plantation in China, ranking top one in the world. To meet the needs of a growing world for social-economic development, environmental improvement, and sustainable development through poplar plantation establishment is a main research interest in the globe. This paper introduced the regionalizing cultivation, key planting clones, and productivities of poplar plantations in China, and summarized the recent 10 years research progress in China in the site quality evaluation, seedling propagation techniques, management patterns, mixed plantation establishment, agroforestry management system, water and nutrient management techniques, site productivity maintenance, and ecological functions of poplar plantation. The potential productivity, general allocation, oriented silviculture, and environmental function study of poplar plantation were also discussed and prospected. The contents of this paper would provide some references for the silviculture and sustainable management of poplar plantation in China. PMID- 19123372 TI - [Toxicological effects of omethoate on leaf photosystem II of cole]. AB - After spraying different concentrations of two brands pesticide omethoate on cole (Brassica campestris L.) leaves, the leaf chlorophyll a fluorescence transients were measured by a Plant Efficiency Analyzer (PEA), and the toxicological effects and rudimental dynamic courses of omethoate on the leaf photosystem II (PS II) were investigated by JIP-test. The results showed that after spraying omethoate except at the concentration of 0.50% , the maximal efficiency of photochemistry (F(v)/F(m)) did not have a remarkable change. However, with increasing omethoate concentration, the minimal fluorescence F(o), maximal fluorescence F(m), relative variable fluorescence at the J-step (V(J)), and electron transport flux perreactive center in PS II (ET(o)/RC) increased remarkably, but psi(o), the efficiency that a trapped exciton in PS ]I moved an electron into the electron transport chain beyond Q(A)-, decreased remarkably. The test two brands of pesticide omethoate had almost alike effects on the PS II of cole, and the residual effect of the pesticide was the strongest at the third day after spraying and petered out from the ninth to twelfth day. The main targets of omethoate on the PS II of cole could be listed as promoting the reduction from Q(A) to Q(A) (-) (increasing of V(J)) and the electron transmission from Q(A) (-) to Q(B) (increasing of ET(o)/RC). PMID- 19123373 TI - [Allelopathic effects of Corallina pilulifera on red tide microalgae Heterosigma akashiwo]. AB - Different concentration methanol-, acetone-, ether-, and chloroform extracts of Corallina pilulifera were used to study their growth inhibitory effects on red tide microalgae Heterosigma akashiwo. The results showed that methanol extract at relatively higher concentrations had the highest growth inhibitory activity and killed all H. akashiwo cells, while the other three kinds of organic solvent extracts had no apparent inhibitory effects, suggesting that the growth inhibitory substances in C. pilulifera had relatively high polarity. The methanol extract was partitioned to petroleum ether phase, ethyl acetate phase, butanol phase, and distilled water phase by liquid-liquid fractionation, and the bioassays on the activity of each fraction were carried out on H. akashiwo. It was found that petroleum ether phase and ethyl acetate phase had strong algicidal effects on H. akashiwo, suggesting that the fatty acids in C. pilulifera tissues might be one of the main allelochemicals. PMID- 19123374 TI - [Investigation of the characteristics of the stagger electrodes dielectric barrier discharge plasmas in chord-wise direction]. AB - The aerodynamic plasma actuator distinguishes itself from others by a set of highly asymmetric electrodes arranged on dielectric. So the plasma produced by the aerodynamic plasma actuator has special characteristics along chord-wise direction. In the present paper the characteristic of the stagger electrodes dielectric barrier discharge plasmas in chord-wise direction was investigated experimentally through spectrometer, infrared imager and laser induced fluorescence system. The mechanisms behind plasma flow control were discussed briefly based on these experimental results. It was found in the experiments that the distributions of light intensity and temperature in chord-wise direction accord with Gaussian distribution. Light intensity and temperature were enhanced by increasing supplied voltage. NO produced by DBD discharge was detected directly by the LIF system. Through numerical simulations, the distributions of electric potential and electric field near the electrodes were determined and the phenomena observed in experiments were explained. Based on these experimental results, the mechanisms behind plasma flow control were ascertained to be the consequence of collisions, temperature increasing and chemical reactions. PMID- 19123375 TI - [Study on determination of plume velocity by passive differential optical absorption spectroscopy]. AB - Differential optical absorption spectroscopy (DOAS) technique has been used to measure various trace gases in the atmosphere by their strongly structured absorption of radiation in the UV and visible spectral range. Passive DOAS using the zenith scattered sunlight as the light source can obtain the continuous column density distribution of air pollutants (such as SO2 and NO2) by scanning the plume emitted from sources on a mobile platform, then with the plume velocity information the total emission value can be ultimately estimated. In practice it is hard to calculate the total emission because there is no efficient way to accurately get the plume velocity which is the most important parameter. Usually the wind speed near ground is used as the actual plume speed, which constitutes the greatest source of uncertainty in the passive DOAS measurements for the total emission calculation. A passive DOAS method for the determination of plume velocity of pollution source was studied in the present paper. Two passive DOAS systems were placed under the plume along the plume transmission direction to observed the scattered sunlight at one fixed sepasation angle, and then the plume velocity was derived from the time delay resulting from the plume moving a certain distance, and also the plume height needed in the plume velocity calculation was measured by the same two passive DOAS systems. Measurement of the plume emitted from a certain power plant was carried out by the two passive DOAS systems and the plume velocities of 3.6 and 5.4 m x s(-1) at two separate moments were derived. The comparison with the wind speed measured at the same time by the single theodolite wind observation method indicates that this optical remote sensing method based on passive DOAS can be used to determine the plume velocity by monitoring the total emission from sources. PMID- 19123376 TI - [Research on cluster-assistant multiple ionization of acetonitrile by intense nanosecond laser]. AB - To further investigate the relation between cluster and multi-charged ions, the photo-ionization of acetonitrile molecular beam carried by helium was studied by time-of-flight mass spectrometry using 25 ns, 532 nm and 1 064 nm Nd-YAG laser with an intensity of 10(10) W x cm(-2). When the effused beam of CH3CN at a pressure of about 11 kPa was irradiated by a 532 nm laser, only a few mono charged ions appeared in the mass spectrum and were clearly resolved, While the pulsed molecular beam seeded in 0.15 MPa He, multi-charged species appeared between H+ and C+, and parent ions containing one or more acetonitrile molecules were also discovered. The intrinsical difference between effused beam and pulsed one was that for the latter it was easier to generate molecule clusters. Adjusting delay time between laser and pulsed valve to shine the laser on different potions of pulsed beam, the change tendency of integral intensity of different multi-charged ions was found to remain similar to that of acetonitrile cluster, and the highest intensity of different ion species appeared at the same 0.75 ms delay time. When the wavelength of laser was changed from 532 nm to 1064 nm, the type of ions and their proportion from effused beam were alike except intensity, but the result of pulsed beam altered obviously. Multi-charged ions showed peak splitting, which characters CE (Coulomb explode). As a powerful tool to understand some chemical physics processes involved, correlation analysis of the time of flight mass spectrum was used. The spectrum of 532 nm was selected owing to its multi-charged ions distinguished easily. The correlations among N(n+) (n = 2-5) and C(n+) (n = 2-4) were all above 0.25, with some even above 0.7, indicating that these ions are highly correlated and probably come from the CE events simultaneously. An electron re-scattering and re-colliding ionization model was proposed to explain the appearances of those multiple charged ions under such low laser intensity. From the theory, the longer wavelength facilitates the energy absorption rate during inverse bremsstrahlung, which leads to the resulting wavelength dependence of the multi-charged atomic ions. PMID- 19123377 TI - [Spectroscopic study of atmospheric pressure argon DC microdischarge]. AB - Microhollow cathode discharge or microdischarge is an efficient method to generate plasma in a high pressure gas. In the present work, the emission spectra were observed in an atmospheric pressure argon direct current microdischarge apparatus, using a stainless steel capillary as the cathode, and a stainless steel mesh as the anode. It was shown that all of the seventeen argon spectral lines arose from electronically excited argon atom 4p-4s transition in the wavelength range of 690-860 nm. The dependences of emission intensity on the discharge current, gas pressure and argon flow rate were investigated. The experimental results show that the emission intensity increased with discharge current from 1 to 6 mA and argon flow rate from 100 to 700 mL x min(-1). The dependence of emission intensity on gas pressure exhibited different characteristics, i.e., spectral signal increased with the gas pressure, but reached the intensity maximum at 13.3 kPa, and decreased afterwards. The argon atom spectral lines 763.51 and 772.42 nm were chosen to measure the electron excitation temperature by the intensity ratio of two spectral lines. The electron excitation temperature was determined to be in the range of 2000 to 2800 K in the atmospheric pressure argon microdischarge. The changes in electron excitation temperature with discharge current, gas pressure and argon flow rate were explored, indicating that the electron excited temperature increased with the discharge current, but decreased when gas flow rate or argon pressure increased. PMID- 19123378 TI - [The development of acetylene on-line monitoring technology based on laser absorption spectrum]. AB - As one of the materials in organic chemical industry, acetylene has been used in many aspects of chemical industry. But acetylene is a very dangerous inflammable and explosive gas, so it needs in-situ monitoring during industrial storage and production. Tunable diode laser absorption spectroscopy (TDLAS) technology has been widely used in atmospheric trace gases detection, because it has a lot of advantageous characteristics, such as high sensitivity, good selectivity, and rapid time response. The distribution characteristics of absorption lines of acetylene in near infrared band were studied, and then the system designing scheme of acetylene on-line monitoring based on near infrared tunable diode laser absorption spectroscopy technology was discussed in detail. Moreover, the system of experiment measurement was set up and the method of signal detection and the algorithm of concentration inversion were studied. In addition, the sample cell with a path length of 10 cm, and the acetylene of different known concentrations were measured. As a result, the detection limit obtained reached 1.46 cm3 x m( 3). Finally the dynamic detection experiment was carried out, and the measurement result is stable and reliable, so the design of the system is practicable through experiment analysis. On-line acetylene leakage monitoring system was developed based on the experiment, and it is suitable for giving a leakage alarm of acetylene during its storage, transportation and use. PMID- 19123379 TI - [Application of laser induced breakdown spectroscopy (LIBS) to microdetection of Au film machining]. AB - Film micro-machining is a core in micro- and nano- technology, micro-electro mechanical systems and photoelectron field. However, it is difficult to control the quality of machining in processing film. In the present paper the authors propose a method to solve this problem by using laser induced breakdown spectroscopy (LIBS). The authors investigated the relation between excitation condition and the quantity of plasma in the process of micro-machining by pulse laser. At low frequency (<20 Hz), when the excitation energy was confirmed, the quantity of plasma was steady and independent of the number of pulses. When the authors used single pulse to excite the film, the quantity of plasma increased with the energy ascension. Based on the quantity of plasma and the species of elements, the authors realized the real-time control of micro-machining become true. PMID- 19123380 TI - [Progress in application of THz-TDS to protein study]. AB - Terahertz time-domain spectroscopy (THz-TDS) is a newly developed terahertz far infrared spectroscopy based on ultra-fast laser technology. Various applications of THz electromagnetic waves have been investigated by researchers in various fields, including life sciences, pharmaceutical, security detection and etc. THz TDS has been proven a promising technique in biological and medical applications. Absorption spectrum in the THz range provides rich information about structure and weak interactions of molecules. It is expected that the THz waves can be applied to probing the interactions between antibody and antigen, drug and organism. Combining the characteristic of THz-TDS and its physical foundation in biomolecular spectroscopy, the paper provides a general review of the achievements of and the latest progresses in THz-TDS technology in the study of molecular structure, dynamics and label-free detection of amino acid, polypeptide and protein. Investigating the structure and dynamics of amino acids and polypeptides in low-frequency range is necessary and fundamental for comprehensive understanding of protein function. The absorption coefficients and refractive indices of different amino acids and peptides were summarized, and the results from different research groups were collected. The combination of absorption coefficient and refractive index enables us to discuss rigorously the dynamics of the total dipole moment of protein system. Biological polymers are expected to exhibit broad vibrational spectral features in the THz range corresponding to functionally relevant, global and subglobal collective modes with periods on the picosecond timescale. Some studies focused on the absorption spectrum of hydrated, dehydrated protein powder and solvated protein in an effort to monitor these collective motions. The results indicated that THz-TDS allows rapid characterization of the dynamical transition without label. However, THz technique is still in its infancy compared with other electromagnetic waves. The problem and the prospect of the application of THz-TDS at this stage are also discussed. PMID- 19123381 TI - [Compensation of backscattering coefficient in Fourier-domain optical coherence tomography]. AB - According to the phenomenon that the intensity of backscattering light decreases dramatically as the detecting depth increases, causing blurred imaging in Fourier domain optical coherence tomography (FDOCT), a compensation method used for different detecting depth was advanced in the present paper. Through compensating the incident power and scattering coefficient of back-scattered light in each layer of the sample, the real gray level of the image was obtained. With the model of multi-layered homogeneous tissue, the main factors influencing image gray were analyzed. Establishing the FDOCT system with multi-layer cover-glass to be the sample, the structure image was obtained. On the basis of cover-glass as the sample, the general formulas for multi-layer compensation were derived. Combining the theoretical compensation and experimental situation, the obtained data were compensated from incident and emergent intensity. The results show that this compensation method is an effective and feasible way to make the deep layer more visible. PMID- 19123382 TI - [The preparation and characterization of 1-D orderly ZnO nanorod arrarys]. AB - Improving on the sealing and high pressure conditions of traditional hydrothermal method, vertical ZnO nanorod arrays were synthesized on indium tin oxide substrate by employing Zn(NO3)2 x 6H2O, (CH2)4N6 as the starting materials in the presence of polyethylenimine(PEI) at ambient pressure and low temperature (92 degrees). Between the substrate and the nanorods, a layer of ZnO flim was prepared as buffer layer and seed layer. The ZnO film was gained by spin-coating zinc acetate solution on indium tin oxide substrate, then annealed at 350 degrees C for 20 min, which can make zinc acetate decompose into zinc oxide. The zinc acetate spin-coating and decomposition procedure was carried out twice to ensure a complete and uniform coverage of ZnO seeds. The second layer was annealed at 500 degrees C for 30 mini Different spin-coating speeds were adopted, one was 2500 r x min(-1), and the other was 5000 r x min(-1). XRD result indicated that the seed layer with 5000 r x min(-1) has better alignment than the layer with 2500 r x min(-1). The aligned seeds with 5000 r x min(-1) show only a (002) reflection, indicating their complete c-axis texturing, whereas the spin-coated seeds give a powder pattern because they rest at all angles on the substrate. SEM result shows that the layer is made up of grains with an the average size of about 30 nm. Well-aligned ZnO nanorod arrays were synthesized by putting the substrate with ZnO seeds into the precursor solutions vertically for one hour. The nanorod arrays were taken out and rinsed with clean ethanol and pure-water for several times, blown dry with a stream of nitrogen, then annealed at 400 degrees C for 30 min in order to wipe off the organic solvent. At room temperature, the SEM and XRD were measured. SEM results indicate that the crystal structure of most of ZnO nanorods is hexagonal wurtzite crystallographic phase structure, mainly vertical to the substrate. ZnO nanorods have good crystallization, the diameter of the rods is around 40 nm, and the length is above micrometer. The XRD results showed the nanorod arrays have (002) and (004) angles, and the (002) is quite strong. Absorption spectra of the nanorod arrays shows ZnO essence absorption and strong ultraviolet absorption, indicating that ZnO has good quality. Optical properties were studied, and the excitation spectra of the nonorod arrays showed a strong and narrow peak at 387 nm with FWHM smaller than 30 nm and a weak blue peak. PMID- 19123383 TI - [Preparation and photoluminescence study of Er3+ : Y2O3 transparent ceramics]. AB - Y2O3 acted as the matrix material, which was doped with different concentrations of Er3+, Er3+ : Y2O3 nanocrystalline powder was prepared by co-precipitation method, and Er3+ : Y2O3 transparent ceramics was fabricated by vacuum sintering at 1700 degrees C, 1 x 10(-3) Pa for 8 h. By using the X-ray diffraction (D/MAX RB), transmission electron microscopy(Philips EM420), automatic logging spectrophotometer(DMR-22), fluorescence analyzer (F-4500) and 980 nm diode laser, the structural, morphological and luminescence properties of the sample were investigated. The results show that Er3+ dissolved completely in the Y2O3 cubic phase, the precursor was amorphous, weak diffraction peaks appeared after calcination at 400 degrees C, and if calcined at 700 degrees C, the precursor turned to pure cubic phase. With increasing the calcining temperature, the diffraction peaks became sharp quickly, and when the calcining temperature reached 1100 degrees C, the diffraction peaks became very sharp, indicating that the grains were very large. The particles of Er+ : Y2O3 is homogeneous and nearly spherical, the average diameter of the particles is in the range of 40-60 nm after being calcined at 1000 degrees C for 2 h. The relative density of Er3+ : Y2O3 transparent ceramics is 99.8%, the transmittance of the Er2+ : Y2O3 transparent ceramics is markedly lower than the single crystal at the short wavelength, but the transmittance is improved noticeably with increasing the wavelength, and the transmittance exceeds 60% at the wavelength of 1200 nm. Excited under the 980 nm diode laser, there are two main up-conversion emission bands, green emission centers at 562 nm and red emission centers at 660 nm, which correspond to (4)S(3/2) / (2)H(11/2) - (4)I(15/2) and (4)F(9/2) - (4)I(15/2) radiative transitions respectively. By changing the doping concentrations of Er3+, the color of up-conversion luminescence can be tuned from green to red gradually. The luminescence intensity is not reinforce with the increase in the concentration, so the doping concentration of Er3+ should not exceed 2%. If the doping concentration of Er3+ exceeds the range, the concentration has very small effect on the improvement of luminescence intensity. PMID- 19123384 TI - [Crystal structures and luminescence property of d10 transition metal complexes]. AB - Five Zn( [II)) and Cd(II) coordination complexes, [Zn(NA)2 (H2O)4](1), [Zn(INA)2 (H2O)4](2), [Zn(2,2'-bi-py)2 (SCN)2](3), [Cd(INA)2 (H2O)4](4) and [Cd(phen)2(NO3)](5) (HNA=nicotinic acid, HINA=isonicotinic acid, 2, 2'-bipy=2,2' bipyridine, phen=1,10-phenanthroline), were synthesized through constant temperature magnetic stirrer or hydrothermal method, and their single-crystal structures were determined by X-ray diffraction. The authors measured the IR, UV Vis-NIR and fluorescence spectra of the complexes and analyzed their photophysical properties. At room temperature in the solid state the five complexes can show strong fluorescence, i.e., lamdamax(em)=362 nm (lamda(ex)=330 nm), (1); lamdamax(em)=424 nm (lamda(ex)=330 nm), (2); lamdamax(em)=442 nm (lamda(ex)=380 nm), (3); lamdamax(em)=424 nm (lamda(ex)=330 nm), (4); lamda(em max)=456 nm (lamda(ex)=360 nm), (5), and complex (5) can emit phosphorescence upon excitation at 360 nm (lamdamax (pl)=546 nm, tau=10 ms). But the organic ligands are different, which lead to the luminescence property of complexes originating from different charge transfer. Compared with the relevant ligands (lamdamax (em)=380 nm, HNA; lamda(em max)=541 nm, 2,2'-bipy), the fluorescence emissions of complex (1) and (3) show a blue-shift which mainly comes from the ILCT (intraligand charge transfer) and at the same time exists L-->M(4S) transfer. The emissions of complex (2) and (4) come from LMCT (ligand-to-metal charge transfer) and show red-shift compared to that of free ligand (lamdamax(em)=337 nm, HINA). For complex (5), the appearance and position of its emission are different from those of the ligand (lamdamax(em)=381 nm, phen), which is attributed to LLCT (ligand-to-ligand charge transfer) and LMCT. PMID- 19123385 TI - [Detecting cerebral hypoxia-ischemia of newborn piglets using spatially-resolved near-infrared spectroscopy]. AB - As a non-invasive technique for measuring tissue oxygenation, near-infrared spectroscopy (NIRS) has increasing applications in detecting cerebral hypoxia ischemia. The authors, introduced the basic principle of the NIRS oximeter developed independently by our group (TSAH-100). The authors achieved the optimal coupling between the probe and the detected cerebral tissue. The present study investigated different regional oxygen saturations of brain (rSO2) measured non invasively by NIRS, arterial blood oxygen saturation (SaO2) measured invasively by blood gas analysis and physiological parameters in newborn pigs with different hypoxia, in order to prove if the non-invasively cerebral rSO2 can indicate cerebral oxygenation status in clinical practice. Using this oximeter, cerebral rSO2 of 28 newborn piglets under different oxygenation status was detected. After mechanical ventilation and inhalation of 8%-17% oxygen for 30 min in the newborn pigs, the pigs were grouped according to the inhalation of oxygen. With the inhalation of 13%-17% oxygen was mild hypoxia group, with 10%-13% was moderate hypoxia group, and with 8%-10% was severe hypoxia group. There were 4 animals in mild hypoxia group, 8 animals in moderate hypoxia group, 12 animals in severe hypoxia group and 4 animals were in the normal control group. The physiological parameters were monitored during the experiment. The SaO2 were invasively measured by blood gas analysis after the experiment. The results indicate that both rSO2 and SaO2 decreased after different degree of hypoxia and there was a good correlation between cerebral rSO2 non-invasively measured by NIRS and SaO2 invasively measured by blood gas analysis (p<0.001). Cerebral rSO2 was also consistent with the degree of hypoxia and the changes in physiological parameters after hypoxia. The arterial pH and the mean arterial pressure (MAP) in the severe hypoxia group was lower than that in the control group (p<0.05). The blood lactic acid in the severe hypoxia group was higher than that in the control group (p<0.05). Thus, the rSO2 can accurately and directly indicate cerebral oxygenation status and can also replace the SaO2 invasively measured by blood gas analysis. Cerebral hypoxia-ischemia can be non-invasively and conveniently diagnosed using NIRS. PMID- 19123386 TI - [Research on the measurement of flue-dust concentration in Vis, IR spectral region]. AB - In the measurement of flue-dust concentration based on the transmission method, the dependent model algorithm was used to invert the flue-dust concentration in the visible, infrared and visible-infrared spectral regions respectively. By the analysis and comparison of the accuracy, linearity and sensitivity of the inversion flue-dust concentration, the optimal spectral region was determined. Meanwhile, the influence of the water droplet with different size distribution and volume concentration was simulated, and a method was proposed which has advantages of simplicity, rapidity, and suitability for on line measurement. Simulation experiments illustrate that the flue-dust concentration can be inverted very well in the visible-infrared spectral region, and it is feasible to use the ratio of the constrained light extinction method to overcome the influence of water droplet. The inverse results all remain satisfactory when 2% stochastic noise is added to the value of the light extinction. PMID- 19123387 TI - [Bio-optical model of total suspended matter based on reflectance in the near infrared wave band for case-II waters]. AB - From August to October, 2006, reflectance spectra were measured in a turbid Case Il waters condition with an ASD FieldSpec spectrometer for a total of 58 samples. Based on the observation of reflectance curves, spectral analysis was carried out over 400-1200 nm. Showing the typical character of Case-II waters, the reflectance values were generally higher than those in other similar studies. Strong backscattering of high concentration total suspended matter (TSM) contributed considerably to the total reflectance spectra in water. Two obvious TSM reflectance peaks were observed in the near infrared wave bands, i.e., 808 and 1067 nm, especially the latter one that was never reported before. The highest correlation coefficient between reflectance and concentrations of TSM existed at 873 nm. Based on the simplification of water inherent optical parameters in the near-infrared wave band, including absorption of TSM, Chlorophyll-a (Chl-a) and chromophoric dissolved organic matter (CDOM), and backscattering of pure water, Chl-a and CDOM, three empirical equations of the bio-optical model using reflectance at 808, 873 and 1067 nm respectively were established to estimate the concentrations of TSM. Compared with linear and exponential models, the bio-optical model showed fairly good performance with comparatively high determination coefficient (r2) and low root mean squared error (RMSE), which confirmed the applicability of the bio-optical model to retrieve concentrations of TSM effectively in turbid Case-II waters. PMID- 19123388 TI - [Method of infrared spectrum on-line pattern recognition of mixed gas distribution based on SVM]. AB - In order to solve the difficulties that the spectrum training data samples of the massive mixed gas cannot be actually obtained, the analysis precision is low and it is not real time online analysis in the analysis of mixed gas component concentration, the support vector machine, a new information processing method, was used in the mixed gas infrared spectrum analysis, and the concept of mixed gas distribution pattern was proposed in the present paper. Based on the thought that the mixed gas distribution pattern recognition is carried out first, and then the analysis work of mixed gas component concentration is done, sixty kinds of mixed gas distribution pattern were determined after investigation and study, and 6000 mixed gas spectrum data samples were used for model training and testing. Sequential minimal optimization algorithm was applied to realize the decrement and the increase of online learning, and finally the model of infrared spectrum online pattern recognition of mixed gas distribution based on SVM was established. The model structure is composed of 2 levels, pattern recognition level and result output level. The pattern recognition level completes the task of mixed gas distribution pattern recognition; while the result output level is composed of 60 SVM calibration models, and it completes the task of mixed gas concentration analysis. Experimental results show that the correct recognition rate of mixture gas distribution pattern is not lower than 98.8%, and that the method can be used for online recognition of mixed gas distribution pattern under the conditions of small samples of a mixed gas, and can add new mixed gas online, and it has the practical application value. PMID- 19123389 TI - [Investigation on baseline correction in pollutant monitoring on-line by NIR spectroscopy]. AB - Spectroscopic data pre-processing is important to spectra scaling and quantitative analysis, and correction of baseline drift caused by variation of measuring condition is critical in NIR spectroscopy analysis in-situ. In the present paper, the NIR absorption spectra of propane and isobutane with different concentration were detected by BRUKER EQUINOX55 spectrometer. The "Derivative smooth" method and the baseline correction method which is accessory of the instrument were theoretically analyzed, and the data processed by the two kinds of method were compared. The results show that the method that is accessory of the instrument can correct steady-state and linear drift, and can gain high SNR, high accuracy; "Derivative-smooth" method has better applicability, can correct non-steady-state and nonlinear drift better, and is a good kinds of pre processing method in analysis in situ. PMID- 19123390 TI - [Target infrared detection in target spray]. AB - Crops in agriculture and forestry are normally planted discretely. The chemical sprayed between crops would cause great waste and serious environment pollution. Therefore realization of the precision spray has great significance. This research discussed the method to realize automatic target detection using infrared detect technology. The infrared can avoid the interference of the visible light effectively and the response speed is very fast. Therefore it can be used to implement non-tough detection. Photoelectric detection systems based on infrared detect technology are normally stable, reliable, low cost, simple structure, and easy to be practically utilized. Therefore it is widely used in the on-line real time detection field. Its key point is to determinate the characteristic wavelength or wave band. The infrared lights emitted from the infrared light emitting diode were irradiated to the detected objects. The reflected infrared lights could be received by the photoelectric device. Then control signal was triggered and automatic target spray was realized. Code division infrared detection circuit was used in the system. Modulated pulse infrared signals using different coding were used in different photodetector units in the built system so as to eliminate the light path interference between different detector units and other light signal interferences. Therefore the interference capacity of the system is high. The test results showed that the automatic target spray equipment set up in the study could detect crop targets automatically. The light wavelength used in the test is 850 nm. The detection range was tunable within 0.1-0.5 m. The least targets detectable distance was less than 0.3 m. PMID- 19123391 TI - [Progress in the study of impact of scattering on stability of quantitative analysis model using near infrared spectroscopy technology and correction methods]. AB - Near infrared spectroscopy technology for inner quality detection and analysis is a rapid and accurate technique. The factors affecting the stability of quantitative analysis models based on NIRS especially the influence of scattering, were investigated. The methods of determination of the parameters of optical properties of sample (scattering coefficient and absorption coefficient) and the interaction between light and sample were summarized. The progress in pretreatment methods to improve the model's stability, such as transformation of standard normal variate (SNV) and multiplication scatter correction (MSC); as well as the multivariate calibration model techniques, such as partial least square (PLS) and primary component analysis regression (PCA), were also described. The model with scatter correction pretreatments and different regression methods were compared to establish a better regression equation by near Infrared reflectance spectroscopy (NIRS). Although pretreatment methods could improve the robustness of model for influence of scattering, new techniques should be employed to eliminate the error of predication under scattering condition when samples are complicated in physical states and are in the large range of concentration. PMID- 19123392 TI - [FTIR spectroscopic studies of facial prosthetic adhesives]. AB - According to the composition of the traditional facial prosthetic adhesives, most of adhesives can be classified into two categories: acrylic polymer-based adhesive and silicone-based adhesive. In previous studies, measurements of various mechanical bond strengths were carried out, whereas the functional groups of the adhesives were evaluated seldom during the adhesion. In the present study the analysis of two facial prosthetic adhesives (Epithane and Secure Adhesive) was carried out by using infrared spectroscopy. Two adhesives in the form of fluid or semisolid were submitted to FTIR spectroscopy, respectively. The results showed that water and ammonia residue volatilized during the solidification of Epithane, and absorption peak reduction of carbonyl was due to the volatilization of acetate vinyl from Secure Adhesive. Similar silicone functional groups both in the silicone-based adhesive and in silicone elastomer could be the key to higher bond strength between silicone elastomer and skin with silicone-based adhesive. The position, shape of main absorption peaks of three adhesives didn't change, which showing that their main chemicals and basic structures didn't change during solidification. PMID- 19123393 TI - [Calibration transfer between two FTNIR spectrophotometers using SVR]. AB - In the present research, a set of maize powder samples was used to study the calibration transfer between two fourier transform near-infrared (FTNIR) spectrophotometers, and a method of moving window support vector regression machines (SVR) was used to correct the differences between the two instruments. Bruker Vector 22/N was referred to as "master" on which the maize protein calibration model was built. Bruker MPA was referred to as "slave" instrument. A transformation matrix was constructed based on the spectra of a sample set (for calibration transfer) measured on both instruments. After transfer, NIR spectra acquired on "slave" will appear as if they were measured on master instrument. The calibration model available for the master can then be used to predict the transformed spectra measured on the slave. The transfer parameters were computed as follows. For wavelength i, the absorbance vector obtained on the master instrument was regressed against the corresponding absorbance matrix of a spectral window obtained on the slave instrument. Method of SVR was used for regression Moving the wavelength i and corresponding window, the transfer parameter for each wavelength can be obtained. For the two FTNIR spectrophotometers, a window size of 31 wavelengths and a subset of 15 transfer samples were chosen to establish the SVR regression model between "master" and "slave". Applying the calibration model to the prediction samples after being corrected by the transfer parameters, a good transfer performance can be achieved. The correlation coefficient (r) is 0.9434, while the relative standard deviation (RSD) is 4.23%. These results suggest that the SVR method can be used to successfully transfer the calibration model for protein of maize developed on a FTNIR spectrophotometer to another. PMID- 19123394 TI - [Passive detection of aeroengine exhaust based on Fourier transform infrared system]. AB - Since the composition and concentration of aeroengine exhaust can reflect the combustion efficiency, they can provide the basis for condition based maintenance, and also the basis for the analysis of environment pollution caused by aeroengine exhaust. So the importance of aeroengine exhaust detection is evident. Up to now, the measurement of aeroengine exhaust is based on sampling analysis which is not convenient and can't meet the detection requirements when an aeroplane is flying-off or flying in the sky. Hence, new methods of exhaust detection must be studied. The passive measurement technology based on Fourier transform infrared spectroscopy (FTIR) was applied to the measurement of aeroengine exhaust in the present paper. At first, the principle of passive measurement based on FTIR was introduced in detail. On this basis, a model algorithm for gas concentration calculation was deduced based on the principle of infrared transmission. Then the feasibility of aeroengine exhaust measurement based on passive FTIR was analyzed, and the passive measurement method of aeroengine exhaust based on FTIR was given. In the end, an experiment of aeroengine exhaust passive measurement was carried out by a FTIR with the type of Tensor 27 produced by BRUKER. Good quality spectra of the exhaust and the background were measured. Based on the model algo rithm of passive measurement, the absorbance spectra of CO and NO were obtained respectively, and the concentrations of CO and NO were figured out. To check up the veracity of this method, a comparison was made with another apparatus. There were only little differences between the results of the two experiments, showing that the passive measurement technology based on FTIR could meet the requirements of aeroengine exhaust detection. PMID- 19123395 TI - [Analysis of near infrared spectra of apple SSC by genetic algorithm optimization]. AB - Genetic algorithm (GA) is an effective method in regions selection applied in building multivariate calibration model based on partial least squares regression. If genetic algorithm is run repeatedly as a block, the optimal solution is obtained faster, the numbers of data used to build calibration model are further reduced, and the prediction precision is further improved. An efficient method named region selecting by genetic algorithms (R-SGA) for building a PLS calibration model of NIR is presented in the present paper, in which each gene of chromosome represents a sub-region. In the R-SGA method, one needs to divide averagely the full spectral band into many sub-regions, and to build a research space with all the combinations of these sub-regions. The FT-NIR spectra were processed by GA after MSC and Savitky-Golay smoothing, a PL S calibration model of NIR wasbuilt by using the optimal combinations of these sub regions. Meanwhile, the full region selecting PLS (FS-PLS) and experiential region selecting PLS (ES-PLS) models were developed using spectra after first order derivative pretreatment. The seven intervals selected by region selecting by R-SGA which contained 434 variables were used as calibration set in GA-PLS. The prediction precision of GA-PLS model was better than FS-PLS and ES-PLS models, with Rc=0.966, RMSEC=0.469, Rv=0.954 and RMSEP=0.797. It was concluded that by using GA technique, in the pretreatment of apple SSC model by PLS, it is possible to optimize data selecting, enhance the precision of prediction and reduce the number of variables of calibration. PMID- 19123396 TI - [Spectroscopic analysis of substituted tetraphenylporphyrin iron, manganese, cobalt, copper and zinc complexes]. AB - A full IR, UV-Vis, FIR, spectroscopic analysis on substituted porphyrin iron, manganese, cobalt, copper and Zinc complexes was performed, and the spectroscopic patterns were found for the metalloporphyrin compounds with various structures. Base on molecular structures theory, the reasons for the spectroscopic patterns were discussed deeply. Considering the inconsistency of the far-infrared spectra of the Fe-Cl axial bonds in chloro-iron tetraphenylporphyrins reported in the literature. Their far-infrared absorptions were investigated by using both theoretical analysis and experimental determination, and the results showing that their vibrational frequencies of the Fe--Cl bonds in different substitution chloro-iron tetraphenylporphyrins were associated with their structures, and were linearly related to the length of Fe--Cl bonds. PMID- 19123397 TI - [Nondestructive measurement of vitamin C in Nanfeng tangerine by visible/near infrared diffuse reflectance spectroscopy]. AB - The present paper reports the study on visible/near-infrared (Vis/NIR) diffuse reflectance spectroscopy for nondestructive measurement of vitamin C in tangerine (Nanfeng, Jiangxi). In the wavelength range of 350-1800 nm, principal component analysis (PCA) and partial least squares (PLS) were used to build mathematic models with respect to pre-processing spectra. The principal components (PCs) and wavelength selection for calibration model were determined based on full cross validation and regression coefficients. The results, based on prediction for 10 unknown samples, are 0.813, 2.112 mg (100 g)(-1) and -0.810 mg (100 g)(-1) for prediction correlation coefficient, root mean square error of prediction, and prediction deviation, respectively. The analysis methods and conclusions of this experiment will provide some reference values for non-destructive measurement of vitamin C and other quality indices in Nanfeng tangerine using Vis/NIR diffuse reflectance spectrometric technique. PMID- 19123398 TI - [Study on component detection of cheese wrapped by polyethylene film based on NIRS]. AB - The possibility of direct determination of fat and protein of wrapped cheese by near infrared spectroscopy was studied. The influences of polyethylene film on the spectra were discussed in order to detect the components of wrapped cheese. And the influences were eliminated using Norris derivation filter pretreatment means. The models for fat and protein of wrapped cheese were calibrated by partial least squares regression (PLS) following eliminating outline, spectral pretreatment, and PLS factors optimization. The best models gave standard errors for calibration of 0.240 and 0.355, standard errors for prediction of 0.326 and 0.219, and correlation coefficients of 0.928 and 0.952 for fat and protein of wrapped cheese, respectively. The results showed no difference from those by non wrapped cheese's models, and were better than wrapped cheese's models without Norris derivation filter pretreatment. Based on the results, it was concluded that near infrared spectroscopy is a reliable, accurate and fast method for non invasive measurement of wrapped cheese fat and protein. PMID- 19123399 TI - [Application and prospect of infrared spectroscopy in environmental science]. AB - With the application of Fourier transform infrared spectrograph and the development of chemical metrology, Fourier transform infrared spectroscopy (FTIR) has become a kind of beneficial tool and means for analyzing samples. Especially with its strong identifying ability, exact determining results and fast response function, the application of FTIR method to environmental science has been popularized gradually. At present, people are launching applied study based on the specific property of FTIR in their corresponding field. Therefore, the present paper mainly introduced the current situation of study, progress and application of infrared spectroscopy in liquid, solid and gas environment state monitoring and qualitative analysis in the nearly past qualitative 10 years. In addition, the future development direction in environmental science is also discussed for its helpfulness in that field. PMID- 19123400 TI - [Effect of the film of gold nanowire arrays on surface enhanced Raman scattering]. AB - Metal nanowire arrays as surface-enhanced Raman scattering substrate, could bring high enhancement effect. According to the electromagnetic enhancement theory, the high enhancement factors come from the strong electromagnetic coupling between nanowires in the arrays. Gold nanowire arrays and gold nanobrushes were prepared by electro-deposition of gold into alumina template coated with gold film, then chemically etching alumina directly or rubbing gold film and chemically etching alumina. SEM showed the nanowires around 60 nm in diameter with length around 600 nm in the nanobrushes. In the gold nanobrushes, local nanowires collect in bundles after complete dissolution of alumina matrix due to the transverse tension during drying in the air. The two kinds of samples are different in structure. But optical transmission measurements of the two samples in the direction perpendicular to the substrate both showed plasma-enhanced absorption around 640-650 nm. The authors choose 4-mercaptopyridine (4-MP) as the probe molecules because 4-MP chemically adsorb on Au nanowires tightly by monolayer and align perpendicular to Au surface. SERS experiments were performend with a 632.8 nm laser. SERS signals come from the back scattering of the incident light propagating perpendicular to the Au substrates. And the Raman spectra were collected at morethan 10 different points on each sample. A strong, locally inhomogeneous surface-enhanced Raman spectrum of 4-MP adsorbed on Au nanobrushes with enhancement factor as high as 1 x 10(6) was observed. At the same time, SERS of 4-MP from the gold nanowire arrays under the same experiment conditions showed the enhancement factor around 10(2). Considering the optical transmission measurements of the two samples, the enhancement factors both come from resonance field enhancement. According to the difference of the two kinds of samples, the higher enhancement factors of gold nanobrushes are related with the gold film. According to the localization theory of electromagnetic field, the field around the upper part of the nanowires of the nanobrushes is far stronger than that of the bottom part and on the film. So the gold film introduced higher enhancement factors in SERS The spectrum of 4-MP also showed that the CT mechanism affects the enhancement factors. PMID- 19123401 TI - [Obtaining aerosol backscattering coefficient using pure rotational Raman spectrum]. AB - Atmospheric aerosol backscattering coefficient ratio can be obtained with the ratio of elastic signal to the total rotational Raman backscattering signal without assuming the ratio of aerosol extinction to backscatter. Generally, the intensity ofpartial rotational Raman spectrum lines instead of the total rotational Raman spectrum lines is measured. The intensity of the total rotational Raman spectrum lines is not dependent on the temperature, but the intensity of the partialrotational Raman spectrumlines is dependent on the temperature. So calculating aerosol backscattering coefficient ratio with the intensity of the partial rotational Raman spectrum lines would lead to an error. In the present paper, the change in the intensity sums of different rotational Raman spectrum lines with temperature was simulated and the errors of aerosol backscattering coefficient ratio derived from them were discussed. A new method was presented for measuring aerosol backscattering coefficient ratio, which needed not to measure the intensity of the total rotational Raman spectrum lines. Aerosol backscattering coefficient ratio could be obtained with the atmospheric temperature and a single rotational Raman spectrum line. Finally, a erosol backscattering coefficient ratio profiles of the atmosphere were acquired with the combined Raman lidar of our lab. The results show that there is no need to assume any relation between aerosol backscattering and extinction or to consider any wavelength calibration to determine the aerosol scattering coefficient. PMID- 19123402 TI - [Study on the pathological uterine tissues with a fiber Raman spectrometry]. AB - The Raman spectrum can reflect the differences in chemical components and molecular structures of tissues and cells, and significant progress has been made in the research on structures, functions and diseases of cells and tissues with Raman spectroscopy. A fiber Raman spectrometer was used to measure the Raman spectra of some uterine malignant, benign, and normal tissues, such as uterine myometrium tissue, uterine myoma tissue, normal endometrium tissue, malignant endometrium tissue and adenomyosis tissue. After having compared the Raman spectrum of pathological tissues with that of the corresponding normal tissues, we observed that the peak referring to Methionine upsilon(C--S) (Met upsilon(C- S)) splits into two peaks in the uterine myoma tissues caused by the vibrations of tryptophan (Trp) and cartotene, which are not present in the normal tissues. There is a peak at 1447 cm(-1) in the endometrium tissues corresponding to CH2- CH3 def, which is one of the characteristic peaks of cancerous tissues. For the adenomyosis tissues, a peak caused by upsilon(C--C) skeletal-alpha helix is obviously weaker than that in normal tissue, and the peak induced by delta(C--O) shifts from 1160 cm(-1) in normal tissues to 1173 cm(-1) in the adenomyosis tissues. Thus, it was demonstrated that the technology of Raman spectroscopy is available for distinguishing different pathological uterine tissues at molecular level. This study is not only helpful on early diagnosis of uterine diseases, but also very crucial for the basic research on uterine diseases. And the Raman spectroscopy technology based on optic-fibers has a potential to evolve into a highly sensitive technology for diagnosis. PMID- 19123403 TI - [Technique of confocal Raman microscopy on erythrocytes]. AB - The technique of confocal Raman scanning microscopy (point scanning, line scanning and 2D scanning) and bright field imaging of living erythrocytes was investigated as a function of different scanning conditions at the excitation wavelength of 514 nm. The biological effect of the 514 nm laser radiation on the erythrocytes was also evaluated, so that a set of proper scanning parameters for different scan modes can be determined to obtain strong enough Raman signal while without damage on the living cells by evaluating the change of Raman spectra and lighted field images of the cells. For the point scanning mode, the laser power at sample is the most important parameter to be adjusted, which normally should be less than 1.5 mW. For the line scanning mode, the laser power at sample and scanning step should be considered at first. Small scanning step means the energy of laser accumulated at a small region, which can easily damage to erythrocytes. Large scanning step can reduce the damage; however the spatial resolution decreases also. It is recommended that scanning step should be more than 0.5 microm and laser power at sample should be less than 0.7 mW. For the 2D scanning mode, besides the laser power at sample, scan step needs to be adjusted, and other scan parameters need to be adjusted properly for reducing the effect of laser on erythrocytes. Large pinhole and relative low temperature of sample are the remedies, which can reduce the effect of laser on erythrocytes. 1.0 microm scanning step, 0.7 mW laser power at sample, 500 microm pinhole and proper low temperature can get better 2D Raman image of erythrocytes. For all scanning modes, if the Raman signal is strong enough, the exposure time can be shortened properly, thus reducing the effect of laser on erythrocytes. The optimization of experiment process is also important for Raman test on living cells. PMID- 19123404 TI - [Investigation of accelerated aging in light and writing age of roller pen inks]. AB - The aging of roller pen ink entries was accelerated in sunlight and ultraviolet, and the extracts of roller pen inks on paper were analyzed by capillary zone electrophoresis, photo diode array (PDA) detector. Scanning range of 190-600 nm was adopted in experiment and the change curves were obtained by the plot of the ratio of peak areas in electrophoretogram vs. accelerated aging. The rule of accelerated aging of roller pen ink was investigated by relative changes in content between different dyes in inks. In addition, the relative writing age of roller pen entries was investigated. PMID- 19123405 TI - [Study on application of Gaussian fitting algorithm to building model of spectral analysis]. AB - In the present paper, Gaussian fitting algorithm is introduced. It is possible to separate some Gaussian peaks in one spectrum and get peak height value, peak position value and other parameters by this algorithm. These Gaussian parameters are used to describe original spectral information and extract spectral feature. The spectral model is optimized and explained by the combination of the Gaussian fitting algorithm and multivariate calibration methods. The relationship between the spectra and the chlorophyll of the corn's leaves is studied in this paper, every spectrum which contains 1551 absorbance data is fitted and separated by three Gaussian peaks, and then 1551 data are converted to 9 Gaussian parameters (approximately 0.58% of the whole original spectral data), and the chlorophyll content is estimated by the Gaussian parameters. This modeling method is fast and accurate for the estimation of a sample. The model of chlorophyll content with the Gaussian fitting algorithm and PLS is built in the range of 400-800 nm, and the experiment results show that the correlation coefficient between the estimated values and the real values is 0.960, and the relative standard deviation is 0.0485; The model of chlorophyll content with the Gaussian fitting algorithm and PCR is built in the same wavelength range, and the experiment results show that the correlation coefficient is 0.962, and the relative standard deviation is 0.048; while the correlation coefficient is 0.957 and the relative standard deviation is 0.051 for the model of PLS without Gaussian fitting algorithm; and the correlation coefficient is 0.919 and the relative standard deviation is 0.077 for the model of PCR without Gaussian fitting algorithm. The reliability of the prediction results shows that Gaussian fitting algorithm is satisfactory for building model of spectral analysis. Compared to the conventional methods, the method of Gaussian fitting algorithm can not only simplify the parameters of models, but also improve the explanation of analysis models. The result of the study shows that it is practical and feasible to apply the Gaussian fitting algorithm to quantitative analysis models. PMID- 19123406 TI - [Study on synthesis and relation between structure and optical properties of stilbene derivatives]. AB - In the present paper, three stilbene derivatives with different substitutions were synthesized by phase-transfer Wittig reaction and Pd(II)-catalyzed heck reaction. The molecular structures were characterized by FTIR, elemental analysis and 1H NMR spectra. The authors investigated the influence of molecular structure on electron absorption spectra and luminescence-emitting properties. The results show that compared with that of compound 3a without substitution with a maximum electron absorption peak at 356 nm, the substitution of CH3 group results in a slightly red-shift of electron absorption peak wavelength of molecules to 358 nm, while the substitution of NO2 group makes molecule possess larger pi electron conjugation and the maximum absorption peak shifts to a longer wavelength region (388 nm). Simultaneously, it was found that the substitution of CHR3 group yields luminescence emission with a peak wavelength at 414 nm and the luminescence emission strength significantly increases in comparison with that of compound 3a without substitution. On the contrary, the substitution of electron-acceptor NO2 group results in the decrease in luminescence emission strength with an emission maximum wavelength at 525 nm. Therefore, the structure of substitution on stilbene molecules has an important effect on their optical properties and this will provide some foundation for the structure design of emitting molecules in future. PMID- 19123407 TI - [Experimental investigation of fluorescence spectra of serum excitated with different wavelength light]. AB - In the present paper the fluorescence spectra of the blood serum excited with different wavelength were measured with the fluorescence photometer RF5301 (SHIMADZU) made in Japan. The relationship between the fluorescence spectra of the serum and the wavelength of the excitation light was studied during the experiment. The experimental results show that the linetype and peak wavelength of the fluorescence spectra of serum excited by ultraviolet radiation with different wavelength are almost the same, and they do not depend on the excitation wavelength. But the fluorescence peak value changes with the excitation wavelength. There are two high intensity emission intervals in the fluorescence spectra. One of these is from about 300 nm to 410 nm, and the other is below 310 nmr The fluorescence spectra are mostly centralized in the first interval, and the wavelengths of fluorescence peaks were found around two locations: one is near 330 nm and the other is near 370 nm. At this time the strife phenomena occur. When the excitation wavelength is about 250 nm or higher, the fluorescence peak only occurs at 330 nm, and the optimal excitation wavelength is 300 nm. While the wavelength is greater than 320 nm, the fluorescence intensity of the first interval begins to fall, while that of the other augments. And at this time the peak of wavelength of fluorescence is 452 nm. This study provides the experimental foundations for advanced study and applications of the characteristics of fluorescence spectrum of blood serum, and also offers the references to the wavelength selection of excitation light in the application of the photo-induced fluorescence spectra diagnostic technology. PMID- 19123408 TI - [Algebraic reconstruction techniques and improvement studied with spectroscopy]. AB - The present article focuses on improving algebraic reconstruction technique (ART) for optical computerized tomography (OCT). With spectroscopy, a series of traditional ARTs were analyzed in detail, then a new improved ART was developed. The new ART was named simple self-correlative algebraic reconstruction technique (SSART). With numerical simulation technique, SSART reconstructing efficiency was demonstrated by reconstructing a simulated field with an opaque object. The simulation result and reconstructing accuracy were analyzed. Many current representative ARTs were studied in order to compare the reconstructing efficiency of the new ART with that of others. These ARTs included basic algebraic reconstruction technique: ART, simultaneous algebraic reconstruction technique: SART, and modified SART: MSART. Each of those was used for reconstructing the same simulated field as that SSART was used to reconstructing. The reconstruction results under the same condition were compared with each other. Reconstruction accuracy of each ART was weighed with three error indexes, mean-square error(MSE), absolute value error (AVE), and peak error (PE). As the results, each error index of the reconstruction result with SSART declined a lot. The MSE of SSART was decreased about 26.6% compared to that of ART at the level of 10(-4) magnitude, and PE 77.6% at the level of 10(-2) magnitude. As far as the field with an opaque object is concerned, SSART improves the reconstruction result much, and increases the reconstruction accuracy a lot. SSART improves ARTs greatly. PMID- 19123409 TI - [Study on the molecular recognization of fungicide of kresoxim-methyl with beta cyclodextrin and its derivatives]. AB - The molecular recognition of fungicide of kresoxim-methyl with beta-cyclodextrin (beta-CD), methyl-beta-cyclodextrin (RAMEB)and hydroxypropyl-beta-cyclodextrin (RAMEB) was investigated by using UV-Vis spectroscopy analysis. The effect of temperature and polarity of solvent on the recognition interaction was studied. The driving force and the possible structure of the inclusion complexes were also discussed. The results presented that they formed inclusion complexes with a stoichiometry of 1:1, and the formation constant of inclusion complexes was in the order of Ku(HP-beta-CD)>(beta-CD)Kp(RAMEB) at 298.15 K. Elevation of the temperature triggered a decrease in stability of the inclusion complexes and the value of K(beta-D) was the biggest at > or =303.15 K. The formation constant reduced sharply with the decreasing polarity of the solvent. The standard molar Gibbs energies, enthalpies and entropies were all negative. All the results indicated that the association of the guest molecule with beta-CD was favored by enthalpy changes, and hydrophobicity and hydrogen bond interaction were main driving forces for the inclusion reaction. Our findings provided an important proof for the use of inclusion complexes of kresoxim-methyl with CDs. PMID- 19123410 TI - [Influence of substituents on Langmuir-Blodgett films and spectral properties of thulium bisphthalocyanine derivatives]. AB - The aggregation and spectral properties of three sandwich thulium bisphthalocyanine compounds in chloroform solution and LB films were investigated by using UV-Vis absorption spectra. The experimental results indicated that the three sandwich thulium bisphthalocyanine molecules formed H-aggregates in the chloroform solution, but the absorption spectra of the three sandwich thulium bisphthalocyanine compounds display strong absorption peaks of monomer when the concentration of the sandwich thulium bisphthalocyanine solutions was 10(-7) mol x L(-1). The addition of substitutent --OC8H17 not only decreased the aggregation behaviors of the sandwich thulium bisphthalocyanine, but also leaded to red-shift of the absorption peaks, and it had a biggish influence on the intensity of absorption spectra. The absorption Soret bands were splitting when the substitutent --OC8H17 was added. The substitutent --OC8H17 had a strong effect on the existence state of bisphthalocyaines in LB films, while TmPc2 and TmPcPc* molecules formed H-aggregates in LB films, however TmPc2* molecule formed T aggregates. In the LB films, the distance between sandwich thulium bisphthalocyanine molecule was shortened, and the interaction of molecules was enhanced, therefore the absorption peaks were red-shifted compared with those in the solution, and the arranging state of sandwich thulium bisphthalocyanine molecule has biggish influence on the peaks of absorption spectra. PMID- 19123411 TI - [Study on inclusion complexes of beta-cyclodextrin with zopiclone]. AB - The inclusion complex formation of zopiclone with beta-yclodextrin was studied by using phase solubility method, and the formation constant for zopiclone-beta cyclodextrin was determined. The solubility of zopiclone and the formation constant were enhanced with the increase in beta-cyclodextrin concentration and pH. The effect of temperature on the reaction was studied through thermodynamics, and the changes in entropy deltaS, enthalpy deltaH and free energy deltaG of the reaction were all negative, suggesting that the inclusion complexation is exothermic and can spontaneously occur by the balance of enthalpy driving and entropy opposing. The inclusion complex of zopiclone and beta-cyclodextrin was prepared by a method of solidgrinding, Stable inclusion complex in solid state was characterized by IR spectra and DTA. Results showed that zopiclone-delta cyclodextrin conclusion complex could be formed, and beta-cyclodextrin is a desirable solubilizer for zopiclone. PMID- 19123412 TI - [Accuracy analysis on a sort of polarized measurement in remote sensing]. AB - Angular error of polarizer in polarimetric measurement is an important element affecting the measurement accuracy of degree of polarization, so angular error of polarizer should be considered in remote sensing of high-accuracy quantitative polarization. Simulation study shows that polarimetric measurement is relative to the polarization state (polarization angle or degree of polarization) of incident light in a specific measurement system of polarization. In the measurement mode of polarizer setting (0 degree, 60 degrees, 120 degrees), there is a maximum error of polarization measurement at the 0 degree or 180 degrees polarization angle while a minimum error at the 30 degrees, 90 degrees and 150 degrees polarization angle; In the measurement mode of polarizer setting (0 degree, 45 degrees, 90 degrees), there is a maximum error of polarization measurement near the 45 degrees polarization angle while a minimum error at the 0 degree, 90 degrees and 135 degrees polarization angle. The larger degree of polarization of incident light often contributes to the bigger measurement error except for incident light with several polarization angles. So the polarization measurement may be evaluated by the average degree of polarizatioo of linearly polarized light introduced in this paper. It is indicated that the measurement mode of polarizer setting (0 degree, 60 degrees, 120 degrees) is better than that of (0 degree, 45 degrees, 90 degrees). PMID- 19123413 TI - [Synthesis, spectral analysis, antibacterial and antitumor activities of Co(II) and Ni(II) ofloxacin complexes]. AB - The interactions of cobalt sulphate and nickel acetate with ofloxacin (ofo, I), a 4-quinoline derivative, were studied. The hydrothermal technique was adopted in this work. The isolated solid complexes were characterized by elemental analysis, infrared spectra, electronic spectra, fluorescence spectra and thermal analysis. The results support the formation of complexes of the formula Co(ofo)2 x 4H2O (II) and Ni(ofo)2 x 4H2O (III). The infrared spectra of the isolated solid complexes suggested that ofo acts as bidentate ligands through one of the oxygen atoms of the carboxylic group and the ring carbonyl oxygen atom. Thermogravimetric (TG) and its differential (DTG) were carried out for the complexes. The data obtained indicated that the thermal decomposition of the two complexes in inert atmosphere proceeded approximately with two main degradation steps. I showed a intense fluorescence in solid state at lamda(ex)/lamda(em) = 365 nm/461 nm, and two complexes displayed weakly similar emission maximum at 470 nm in the powder samples at ambient temperature, while the emission of II and III may be mainly originated from the intraligand excited states of I. I, II and III were assayed against two kinds of gram-positive and two kinds of gram-negative bacteria by in vitro doubling dilutions method. The results indicated that II and III have the similar minimal inhibitory concentration (MIC) as the I against S. Aureus, M. Lutens, E. Coli and P. Aeruginosa. The inhibitory effect of I, II and III on leukemia HL-60 cell line has been measured by using MTT (Methyl-Thiazol Tetrozolium) method and that on liver cancer BEL-7402 cell line measured by SRB (Sulphurhodamin B) method. The results indicated that the complexes in the high concentration have inhibitory effect on HL-60 cell line. PMID- 19123414 TI - [Simultaneous resolution of overlapped spectra of three kinds of organic compounds using a wavelet packet transform-based generalized regression neural network]. AB - A wavelet packet transform-based generalized regression neural network (WPTGRNN) was developed to perform simultaneous spectrophotometric determination of p nitroaniline, alpha-naphthylamine and benzidine. This method combines wavelet packet transform (WPT) with generalized regression neural network (GRNN) for improving the quality of noise removal and enhancing the ability of prediction. Wavelet packet representations of signals provided a local time-frequency description and separation ability between information and noise. The quality of noise removal can be further improved by using best-basis algorithm and thresholding operation. Generalized regression neural network (GRNN) was applied for overcoming the convergence problem encountered in back propagation training and facilitating nonlinear calculation. The GRNN is also advantageous in that the training process is much faster and without making any assumption about the form of the prediction model. By optimization, the wavelet function, decomposition level and smoothing factor of GRNN were selected. The partial least squares (PLS) method was used for comparative study. PLS method uses both the response and concentration information to enhance its ability of prediction. Three programs, PWPTGRNN, PGRNN and PPLS, were designed to perform relative calculations. Experimental results showed WPTGRNN method to be successful and better than others. Compared with GRNN method, the relative standard errors of all components between the actual and estimated values of mass concentration for WPTGRNN method decreased from 4.0% to 2.3%. Aniline type compounds are widely applied in industries such as chemistry, printing and pharmacy, and are one of the most important raw materials for synthetic medicine, dye, insecticides, polymer and explosives. Aniline-type compounds are highly poisonous, and can also cause cancer. Simultaneous determinations of aniline-type compounds are very important in environmental and industrial analysis. PMID- 19123415 TI - [Principal components analysis with sensation network applied in the recognition of medicine spectrum]. AB - On the basis of classical theory about spectral analysis, the present article used the method of principal component analysis to get the specificity of 83 ultraviolet absorption spectra from mammary gland patient pathology pieces of 83 cases. The authors chose 44 principal component data as training samples and the rest 39 as testing samples. After training discrete and continual sensation network, the authors found that the recognition rate of cancer was only 43.3% and the recognition of noncancerous one was 38.7% when using the discrete sensation network However, because fuzzy-mathematics was introduced to the continual sensation network and the output value of this model was expanded to [0,1], the recognition rate of cancer reached 83.6% and that of noncancerous one was 76.3% when using this model. PMID- 19123416 TI - [Distribution of Pb and Zn in transgenic metallothionein tobacco]. AB - Transgenic metallothionein (MT) plant can clear the heavy metals from soil and environment, but the distribution of metals in plants has not been studied systematically. The Pb and Zn contents in different parts of transgenic MT tobacco plant of sixth generation and traditional plant (same culture variety as control) were analyzed. The Pb and Zn contents in total transgenic plant were 21.8% and 27.2% higher than control, respectively. The distribution of Pb and Zn in different organs varied in these two types of plants. The Pb and Zn contents in old leaves, stem and root in transgenic plants were significantly higher than those in wild type tobacco, while there was no significant difference in young leaves. The Pb contents in old leaves and root were 30.2% and 47.8% higher than those in the control, and the Zn contents in old leaves, stem and root were 4.7%, 29.2% and 21.6% higher than those in the control. These data showed that Pb was accumulated in old leaves and root easily, while Zn was accumulated in old leaves and stem easily. PMID- 19123417 TI - [Analysis and estimation of the phosphorus content in cucumber leaf in greenhouse by spectroscopy]. AB - A handheld spectroradiometer was used to measure the spectral reflectance of the crop with the measurable range from 325 nm to 1075 nm. Since the first derivative of the spectra can well eliminate spectral error, it was calculated for each spectrum. The cucumber leaves were also sampled and the phosphorus content was measured for each sample with chemical method. First, the correlation between the phosphorus content of the cucumber leaf and the spectral reflectance was analyzed but high coefficient was not obtained. It was shown that there is not high linear relation between those. Then, the analysis was conducted between the phosphorus content of the cucumber leaf and the first derivative of spectrum for each sample. The coefficients were improved. However, it was not high enough to establish an estimation model. It shows that non-linear model is needed to estimate the phosphorus content of the crop leaf based on spectral reflectance. Artificial neural network (ANN) and support vector machine (SVM), the modern ealgorithm for modeling and estimating, were used to establish the nonlinear models. From stepwise multi-regression, four wavelengths, 978, 920, 737 and 458 nm, were selected as modeling wavebands. For the Artificial Neural Network (ANN) model, the data of spectral reflectance in the four wavebands were taken as the input and the phosphorus content was taken as the output. And the number of the neurons in the middle layer, the learning rate, and the learning error were set as 25, 0.05, and 0.001, respectively. The calibration accuracy of the model was 0.995, and the validation accuracy reached to 0.712. For the Support Vector Machine (SVM) model, the selected kernel function was anova, and the penalty parameter C and the linear epsilon-insensitive loss function were set as 100 and 0.00001, respectively. The calibration accuracy of the model was closed to 1, and the validation accuracy reached to 0.754. It can be concluded that both nonlinear models are practical. PMID- 19123418 TI - [Study on fluorescence spectra of Chinese rice wine]. AB - Chinese rice wine has a history of more than 5000 years, only in China, hich is called "Nation Wine", and Shao Xing Chinese rice wine is a representative of it. The fluorescence spectra of 3 kinds of Shao Xing Chinese rice wine with different storage age induced by suitable UV-light were obtained in the present paper through experiment. The authors compared and analyzed them comprehensively by using three dimensional contour and two dimensional fluorescence technique, and found that their best induced light was all in the vicinity of 370 nm. The peak wavelength of 3 year storage age wine was 504 nm, while 488 nm and 505 nm were for 5 and 8 year storage age wine respectively. The emitted fluorescence existed in 400 to 680 nm, which had wide peak. More over, the emitting mechanism of Shao Xing Chinese rice wine was discussed. The authors also have explained the similarity and difference of fluorescence spectra among the 3 kinds of wine. The reason for wide fluorescence peak was also analyzed. This investigation contributes to the study of the characteristic, year detection and food safety of Chinese rice wine. PMID- 19123419 TI - [Preparation of Nd2O3 nanoparticles by ion exchange resin homogenous precipitation method]. AB - In the present paper, homogenous precipitation with Nd(NO)3.6H2O and (NH)2CzO4.2H2O, a strong base anion exchange resin and strong acidic cation exchange resin, respectively, and subsequent calcination of precursors at 700 C were used to synthesize Nd2O3 nanoparticles. In the precipitation from homogeneous solution, the relative supersaturation is always low and the local reagent excesses do not occur because the precipitating agent appears homogeneously throughout the entire solution. In general, homogeneous precipitates are better suited for analysis than precipitates formed by direct addition of a reagent. The concentration of reactant, reaction temperature and time are important factors which dominate the morphology and structure size of the Nd2O3 nanoparticles. Assay results showed that between 40 and 50 degrees C, when the ion exchange resin was added tardily with continuous stirring and mixing, the precursor could be obtained after being filtered, washed, and dried at 80 degrees C. The Nd2O3 Sample was obtained via calcination of the precursor at 700 degrees C for 1.5 h. The morphology, size and distribution of the precursors and samples were characterized by TG-DTA, XRD, TEM, HR-TEM, SAED and BET techniques. The forming mechanism of homogenous precipitation was discussed detailedly. The results indicated that the Nd2O3 nanocrystal prepared with the present method belonged to hexagonal crystal system, and its average diameter was 26-32 nm with better decentralization. The electron diffraction pattern which had a clear and regular lattice of Nd2O3 confirmed that the particles were monocrystals. The specific surface area of sample a and sample b was 42. 57 and 29.43 m2 x g(-1), respectively. The ion exchange resin homogenous precipitation method has such advantages as low cost, environmentally benign innocuity of the starting reagents, the simplicity of the synthetic route, easy recycling of ion exchange resin and so on, making it a promising method to be scaled up for industrial production. PMID- 19123420 TI - [Determination of the retrieval arithmetic of aerosol size distribution measured by DOAS]. AB - Atmospheric aerosol is not only an important factor for the change in global climate, but also a polluting matter. Moreover, aerosol plays a main role in chemical reaction of polluting gases. Determination of aerosol has become an important re- search in the study of atmospheric environment. Differential optical absorption spectroscopy (DOAS) is a very useful technique that allows quantitative measurement of atmospheric trace gas concentrations based on their fingerprint absorption. It also can be used to retrieve aerosol extinction coefficient. In the present work, the method of determination of aerosol size distribution measured by flash DOAS is described, and the arithmetic based on Monte-Carlo is the emphasis. By comparison with the concentration of PM10, visibility and Angstrom wavelength exponent, a good correlation can be found. Application of DOAS in aerosol field not only provides a novel method for aerosol detection, but also extends the field of application of DOAS technology. Especially, aerosol DOAS plays an important role in the study of atmospheric chemistry. PMID- 19123421 TI - [Determination of trace elements in waste beer yeasts by ICP-MS with microwave digestion]. AB - The waste beer yeast has rich nutritional compositions and is widely used in food, medical and forage industries. The security of the yeast plays an important role in everyone's daily life. But the yeast contanining microamount of lead, cadmium, chromium, arsenic and other harmful metals is endangering human health. A new method was developed for the direct determination of eight elements, namely copper, lead, zinc, iron, manganese, cadmium, chromium and arsenic in waste beer yeast by inductively coupled plasma-mass spectrometry (ICP-MS) with microwave digestion. The parameters of plasma system, mass system, vacuum system and spectrometer system were optimized. The spectral interferences were eliminated by selecting alternation analytical isotopes of 65Cu, 208Pb, 66Zn, 57Fe, 55Mn, 114Cd, 52Cr and 5As, and the internal standards of Rh was selected to compensate the drift of analytical signals. The samples were digested with concentrated nitric acid-hydrogen peroxide (2:1) mixed solution more rapidly and more effectively. The effects of the type of mixed acid , the volume of digesting solution, heating time, and heating power were investigated in detail. In the closed system, the complete digestion was performed using 4 mL HNO3 and 2mL H2O2 for 2.0 min at 0.5 MPa, 3 min at 1.0 MPa and 5 min at 1.5 MPa. The detection limits of these eight elements were 0.013-0.122 microg x L(-1). The relative standard deviation (RSD) was 0.94%-3.26% (n=9), and the addition standard recovery was 98.4%-102.6% for all elements. The proposed method has been applied to the determination of trace elements of Cu, Pb, Zn, Fe, Mn, Cd, Cr and As in waste beer yeast samples with satisfactory results. The determination results indicated that the content of trace elements of Cu, Pb, Cd and As in waste beer yeast samples are significantly low. PMID- 19123422 TI - [Application of ICP-MS to determination of heavy metal content of heavy metals in two kinds of N fertilizer]. AB - Environmental safety has been the focus worldwide, where involved are the pollutions of heavy metals, pesticides and persistent organic pollutants. Fertilizer has become one of the polluting sources of heavy metals, which are very deleterious to human health and environmental safety. Heavy metals are difficult to metabolize in human body and very harmful, so research on the pollution of heavy metals is considered increasingly important. The pollution sources of heavy metals include waste residue, waste water and exhaust gas from industry and automobile, and garbage from human life. The heavy metals in fertilizer can endanger the human body by the crop containing heavy metals. Two kinds of nitrogen fertilizer were analyzed in terms of the content of heavy metals by ICP-MS, and the results showed that the content of 10 kinds of heavy metals (Al, Ti, Cr, Ni, Cu, Zn, As, Cd, Hg and Pb) in (NH4)2SO4 was 1345.13, 35.12, 2539.27, 287.26, 674.05, 270.79, 42.54, 22.13, 27.20 and 123.87 ng x g(-1) respectively; and in CO(NH2)2 it is 71.59, 5.36, 1167.71, 188.60, 7.46, 64.45, 10.55, 0.00, 0.09 and 3.71 ng x g(-1) respectively. All the data showed that CO(NH2)2 contained much less heavy metals than (NH4)2SO4, so we should select CO(NH2)2 as the nitrogen fertilizer in agricultural production. PMID- 19123423 TI - [Application of ICP-MS to detecting ten kinds of heavy metals in KCl fertilizer]. AB - With the rapid development of society, more and more attention has been focused on environmental safety, especially on the pollutions of heavy metals, pesticides, persistent organic pollutants and deleterious microorganism. Heavy metals are difficult to metabolize in human body are quite harmful, so research on the pollution of heavy metals is increasingly important. There are many pollution sources of heavy metals, including waste residue, waste water and exhaust gas from industry and automobile, and garbage from human life. The contents of 10 kinds of heavy metals (Cr, Ni, Cu, As, Cd, Sn, Sb, Hg, Tl and Pb) in potassium fertilizer (KCl) from Russia were analyzed by ICP-MS. The results showed that potassium fertilizer (KCl) contained less heavy metals than organic inorganic compound fertilizer; the content of heavy metals Cr, Ni, Cu, As, Cd, Sn, Sb, Hg, Tl and Pb is 0.00, 65.54, 238.85, 190.60, 0.98, 14.98, 2.97, 10.04, 1.28 and 97.42 ng x g(-1), respectively, which accords with the correlative standards. All the data showed that if potassium fertilizer (KCl) is manufactured through normal channel, the content of heavy metals should be little and safe. PMID- 19123424 TI - [Determination of lead in microemulsified rapeseed oil and bio-diesel oil by GFAAS]. AB - Bio-diesel oil has attracted much attention as a substitutable energy sources for its renewable and eco-friendly property. However, problems of lead contamination in fuel are also emphasized increasingly at present. So it was of quite significance to determine the contents of lead in bio-diesel oil and its raw material rapeseed oil. An effective method was developed for the rapid determination of lead in rapeseed oil and bio-diesel oil by graphite furnace atomic absorption spectrometry (GFAAS) after their stabilization as microemulsions. In this research work, polyethyleneglycol octyl phenyl ether and n-butanol were used for emulsifier and auxiliary emulsifying agent, respectively. For Pb, efficient thermal stabilization was obtained using NH4H2PO4 as matrix modifier. Sample stabilization was necessary because of evident analyte losses that occurred immediately after sampling. Excellent long-term sample stabilization and the influence of the microemulsion composition on the GFAAS response were observed by mixing different organic solvents. The ashing and atomization temperature and ramp rate influenced the sensitivity obtained for Ph. Take this into account, the optimum conditions of the graphite furnace atomic absorption spectrometric determination of Pb in rapeseed oil and bio-diesel oil samples were investigated. The results showed that the microemulsion was quite stable when the value of V(20% polyethyleneglycol octyl phenyl ether), V(n butanol), V(oil) and V(water) was 0.1: 8.9: 0.5: 0.5, without matrix interference effect. The determination limit of the proposed method was 126.2 microg x L(-1) for Pb, comfortably below the values found in the analyzed samples. The recoveries were from 81.8% to 109.0%, which performed using the addition of different concentrations of lead to bio-diesel oil, rapeseed oil and petrochemical diesel samples. The relative standard deviation of determination was 5.84%. This work showed the great efficiency of the microemulsion, indicating that it is possible to extract lead from the oil phase. The method was applied to the determination of lead in oil samples with satisfactory results. PMID- 19123425 TI - [Determination of arsenic species in bunge pricklyash seed by HG-AFS]. AB - Some of traditional Chinese medicine (TCM) contain arsenide, such as realgar. The total amount of arsenic in the TCM exceeds the limits according to related regulations. But the roles of arsenic in TCM or its side-effects depend on its species existing in those therapies, not on the total amount of arsenic. Therefore, in recent years, the analysis of arsenic in TCM focuses on the species of arsenic. The present paper combines the method of extracting chemical composition from bunge pricklyash seed and analyzing the contents and the speciation of arsenic of bunge pricklyash seed (BPS) in different habitat by atomic fluorescence spectrometry (AFS-3100). By using 0.45 microm filter membrane and mixed solvent, two different species of the decoction of BPS were extracted, named the suspended and the soluble, and by using ion-exchange resin, the soluble arsenic could be separated as As(III), As(V), MMA and DMA. The authors established the segregation analysis methods for those species. The results shows that BPS in different habitat all contain arsenic, mostly as the species of inorganic arsenic, and the contents of As(V) are higher than As(III). The lixiviating rate of arsenic is highest with the solvent of methanol-water (4/1), reaching 53.7%-62.3%. The recovery of the method was 96.0%-101.3% and the relative standard deviation was less than 2.66%. The results show that the sensitivity and reproducibility of this method were excellent, and provided theoretical basis for further development of BPS PMID- 19123427 TI - [Preparation and characteristics of a novel Cu(II)-imprinted epoxy resin-based monolithic polymer]. AB - A novel Cu(II)-imprinted monolithic polymer was prepared by step-polymerization reaction of epoxy resin. A homogenous mixture of epoxy resin, diethylenetriamine as curing agent, copper ion as target molecule and polyethylene glycol as pore forming reagent was poured into a glass-tube and polymerized under controlled reaction temperature and time. The PEG and copper ions were all removed from the solid product by deionized water and strong acid, respectively. The Cu(II) imprinted epoxy resin-based monolithic porous polymer was successfully obtained, which was used as solid-phase extraction matrix for preconcentration and separation of trace copper ions in aqueous solution for determination by atomic adsorption spectrometry. The effect of pH on adsorption, maximum adsorption capacity, selectivity, analytical precision and availability was investigated in details by a series of experiments. Under the optimized enrichment conditions, i.e., pH 5.0-9.0 of medium and flow rate below 5.0 mL x min(-1), the recovery of Cu(II) ions in the aqueous solution could be over 95%. The maximum adsorption capacity is 126 mg Cu (II) x g(-1) dried resin. The selective recognition ability of Cu(II))-imprinted monolithic column is evidently preponderant over the non imprinted materials. The up-loaded column could be completely eluted with 1.0 mol x L(-1) HNO3 and flow rate below 2.0 mL x min(-1). The proposed procedure was applied to the analysis of a mixed sample with satisfied result. PMID- 19123426 TI - [Principal component and cluster analysis of trace elements in Chinese herb Salvia miltiorrhiza and its relative species]. AB - The contents of trace elements, including copper, zinc, iron, magnesium, calcium, chromium, lead, molybdenum, manganese, and cadmium in Chinese traditional herb S. miltiorrhiza and its relative species such as S. miltiorrhiza f. alba, S. evansiana, S. yunnanensis, S. przewarlskii, S. cavaleriei, S. cavaleriei var. simplicifolia, S. roborowskii, S. omeiana, S. tricuspis, S. brevilabra and S. cynica were determined by atomic absorption spectroscopy. The principal components analysis of SPSS was applied to the study of characteristic elements in S. miltiorrhiza and its relative species. Three principal components which accounted for 79.3% of total variance were extracted from the original data. The first factor accounted for 49.6% of the total variance, which means that iron, manganese, copper, zinc, cadmium and lead are the characteristic elements in S. miltiorrhiza and its relative species. The results of cluster analysis show that the samples could cluster reasonably into two groups. The samples of S. miltiorrhiza from different regions were classified into one group except S. miltiorrhiza from Zhejiang and S. miltiorrhiza f. alba. The other Salvia species were classified into another group except S. cavaleriei. The S. miltiorrhiza and other Salvia species can be distinguished by this method, whose accuracy of classification is 90%. The cluster analysis based on the contents of trace element in S. miltiorrhiza and its relative species provided a quick, accurate and simple method for authentication of herb Salvia miltiorrhiza. PMID- 19123428 TI - [Application of direct demodulation technique in analysis of ground verification test of x-ray spectrometer for Chang'e mission]. AB - An X-ray fluorescence imaging spectrometer based on silicon PIN photodiodes was designed and constructed for the Chang'E mission, the first lunar spacecraft, and will be in operation at a 200 km circular lunar orbit with one year lifetime. The X-ray fluorescence spectrometer consists of two silicon PIN photodiodes modules, each holds two low energy detector units to analyze the distribution of useful elements and to estimate the abundance on the moon, which is one of the objectives of the X-ray fluorescence spectrometer experiment. The low energy detector unit is 25 mm2, 500 microm thick, with the energy band of 1-10 keV, and energy resolution of: approximately 5% at 5.9 keV. The ground verification tests of the X-ray spectrometer for Chang' E mission were introduced in the present paper. Taking the energy response matrix of the spectrometer as the foundation and using the direct demodulation technique and fundamental parameter method, the authors performed some quantitative and qualitative analysis of these scientific data which came from the ground verification tests, especially for Mg, Al and Si elements. PMID- 19123429 TI - [Design of hyperspectral imaging system based on LCTF]. AB - A new compact lightweight imaging system for hyperspectral imaging is described. The system can be thought of as the substitute for traditional mechanical filter wheel sensor. The system is based on different techniques. It uses an electronic controlled LCTF(liquid crystal tunable filter) which provided rapid and vibrationless selection of any wavelength in the visible to IR range. The imaging system consisted of an optic lens, a CRI VariSpec LCTF and a Dalsa 1M30 camera. First the outline of this system setup is presented, then the optics designed is introduced, next the working principle of LCTF is described in details. A field experiment with the imaging system loaded on an airship was carried out and collected hyperspectral solid image. The images obtained had higher spectral and spatial resolution. Some parts of the 540-600 nm components of the 16-band image cube were also shown. Finally, the data acquired were rough processed to get reflection spectrum(from 420 to 720 nm) of three targets. It is concluded that the experiment has proved that the imaging system is effective in obtaining hyperspectral data. The image captured by the system can be applied to spectral estimation, spectra based classification and spectral based analysis. PMID- 19123430 TI - [Research on calibration transfer of NIR filter spectrophotometer]. AB - Calibration transfer is an important issue to building up universal and comparable performance of spectrometer data in near infrared spectral analysis technology. Methods of slope/bias correction, direct standardization (DS), and target factor analysis (TFA) were used for the calibration transfer among five NIR filter spectrophotometers using maize as the samples. The effects of three calibration transfer methods were compared. The DS method has the best performance. The average calibration transfer difference of DS is 7.01%. This study also relates to the dependence of calibration transfer on the number of standardization samples. It was proven by experiment that the results of calibration transfer will be better as the number of samples is increased and will be generally stable when there are twenty standardization samples. However, the effect of calibration transfer attained by DS is degraded sharply when the number of standardization samples decreases to be below twenty. Moreover, slope/bias and TFA are not sensitive to the number of standardization samples. PMID- 19123431 TI - Sleep and allergic rhinitis. AB - Patients with chronic diseases, including chronic respiratory diseases, usually have considerably impaired sleep quality that may increase the frequency of exacerbations and severity of symptoms, lead to difficulty in patient management, and reduce quality of life (QOL). During the last few decades, several studies have shown that, in addition to the classic signs of sneezing, nasal itching, rhinorrhea, and nasal obstruction, allergic rhinitis has an important impact on the QOL of adults and children. In 2001, the ARIA (Allergic Rhinitis and its Impact on Asthma) report based its new severity classification on the impact of rhinitis on QOL, with the inclusion of sleep disturbances. Thus, allergic rhinitis patients may also suffer from sleep disorders, emotional problems, as well as impairment in daily activities and social functioning. Given that sleep is fundamental for physical and mental health, the present document reviews the methods and questionnaires used to assess the quality of sleep, the importance of sleep in allergic rhinitis, impairment and improvement of sleep in allergic rhinitis by using medications (antihistamines, topical nasal corticosteroids, nasal decongestants, antileukotrienes) and, finally, the relationship between the sleep apnea syndrome with allergic rhinitis and its treatment. PMID- 19123432 TI - Obesity and asthma. AB - Asthma and obesity have a considerable impact on public health and their prevalence has increased in recent years. Numerous studies have linked these disorders. Most prospective studies show that obesity is a risk factor for asthma and have found a positive correlation between baseline body mass index and the subsequent development of asthma. Furthermore, several studies suggest that whereas weight gain increases the risk of asthma, weight loss improves the course of the illness. Different factors could explain this association. Obesity is capable of reducing pulmonary compliance, lung volumes, and the diameter of peripheral respiratory airways as well as affecting the volume of blood in the lungs and the ventilation-perfusion relationship. Furthermore, the increase in the normal functioning of adipose tissue in obese subjects leads to a systemic proinflammatory state, which produces a rise in the serum concentrations of several cytokines, the soluble fractions of their receptors, and chemokines. Many of these mediators are synthesized and secreted by cells from adipose tissue and receive the generic name of adipokines, including IL-6, IL-10, eotaxin, tumor necrosis factor-alpha, transforming growth factors-beta1, C-reactive protein, leptin, and adiponectin. Finally, specific regions of the human genome related to both asthma and obesity have been identified. Most studies point out that obesity is capable of increasing the prevalence and incidence of asthma, although this effect appears to be modest. The treatment of obese asthmatics must include a weight control program. PMID- 19123433 TI - Adaptation and validation of the Spanish version of the Chronic Urticaria Quality of Life Questionnaire (CU-Q2oL). AB - OBJECTIVES: The aim of this study was to develop a Spanish version of the Chronic Urticaria Quality of Life Questionnaire (CU-Q2oL) and to test its acceptability, reliability, validity, and sensitivity to change. METHODS: Forward and back translation by bilingual translators followed by pilot testing in patients with urticaria was used to adapt the questionnaire. The Spanish version of the CU-Q2oL was self-administered alongside the Skindex-29 in an observational, longitudinal, multicenter study. Feasibility was assessed by analyzing missing responses and ceiling and floor effects. Reliability was tested by examining internal consistency (Cronbach alpha). Construct validity was analyzed by examining convergent and discriminant validity with the Skindex-29 and by evaluating the ability of the CU-Q2oL to discriminate between patients according to a clinical classification of severity. Sensitivity to change was analyzed in a subgroup of patients who completed a second visit 4 weeks after baseline. RESULTS: A total of 695 patients were included in the analysis. Mean (SD) age was 42.4 (15.0) years and 62.1% of the sample was female. All of the items on the CU-Q2oL were answered by 91.9% of the sample. Over 15% of patients scored at the floor (best possible health) on 5 of the 6 dimensions. Cronbach alpha coefficients were > 0.80 for all dimensions of the CU-Q2oL, and 0.86 for the overall score. Construct validity was supported by correlations between the CU-Q2oL and the Skindex-29, which generally fulfilled hypotheses, and by the questionnaire's ability to discriminate between groups with different severities of urticaria.The questionnaire was sensitive to change, with an effect size of 1.0 for the overall score in patients reporting an improvement on the health transition scale. CONCLUSIONS: The Spanish version of the CU-Q2oL has shown satisfactory reliability, validity, and sensitivity to change. It is suitable for use as an outcome measure for chronic urticaria patients in clinical and research settings. PMID- 19123434 TI - Applicability and reproducibility of biomarkers for the evaluation of anti inflammatory therapy in allergic rhinitis. AB - BACKGROUND: We aimed to study the reproducibility of several biomarkers of allergic rhinitis to investigate their potential as outcome measures in clinical intervention trials. Furthermore, we investigated the kinetics of the biomarkers studied in nasal lavage and brush material following a placebo-controlled nasal allergen challenge. METHODS: We performed a skin prick test and measured serum specific immunoglobulin (Ig) E levels and inflammatory biomarkers in nasal lavage and brush material in 20 patients with allergic rhinitis on 2 separate days (washout, 14-21 days). The patients were then randomly assigned to undergo an intranasal challenge with a relevant allergen (n=10) or diluent (n=10) in order to assess the kinetics of several biomarkers of allergic airway inflammation in nasal lavage and brush samples. RESULTS: Baseline serum IgE levels and skin wheal sizes were highly reproducible measurements, with a coefficient of variation (CV) of 13.4% and 18.2%, respectively. This was not the case with the majority of inflammatory biomarkers, whose CV varied considerably (range, 6.1%-224.1%). The nasal allergen challenge induced an increase in composite symptom scores in all patients. Compared to placebo, tryptase (P=.004), eosinophilic cationic protein (ECP) (P=.03) and alpha2-macroglobulin (P=.002) were increased in nasal lavage at 20 minutes post allergen. Nasal lavage ECP levels and nasal brush eosinophils were still significantly increased at 7 hours (P=.03 and P=.04), but all statistical significance had been lost at 24 hours post challenge. CONCLUSION: Serum specific IgE assays and skin prick tests exhibited good reproducibility in patients with clinically stable allergic rhinitis. We were also able to investigate the kinetics of allergen-induced upper airway inflammatory markers in nasal lavage and brush material. Hence, nasal allergen challenge, when used in combination with nasal lavage and brush sampling, is a suitable research tool for early drug development. PMID- 19123435 TI - Immunoquantitative measurement of soybean aeroallergen emissions at industrial sites. AB - BACKGROUND: Asthma attacks and mortality due to inhalation of soybean antigens in Barcelona, Spain have been well documented. A new control scheme was adopted in the city to avoid the emission and dispersion of soybean dust into the atmosphere during unloading. We studied soybean allergen emission during unloading and at 3 industrial sites and compared the results obtained. METHODS: Over a period of 31 months, 628 paired air samples from 3 plants (A, B, C) involved in soybean manipulation in Barcelona harbor were collected. Samples were analyzed by a radiometric competitive inhibition assay (RCIA) and the enzyme-linked immunosorbent assay (ELISA). A Bland-Altman plot was used to compare the soybean concentrations measured by each assay. RESULTS: The median values for the 628 samples were 5535 U/m3 (range, 370-18,416,751) for the RCIA and 9955 U/m3 (range, 400-22,349,059) for the ELISA. Plant A had the lowest emission levels and the lowest Spearman rank correlation coefficient (0.409). The correlation coefficients were 0.747 and 0.794 for plants B and C. Soybean aeroallergen concentrations differed by plant. The highest variability in values was seen for plant A, which had the lowest allergen concentrations. CONCLUSIONS: The competitive assays described are useful tools for the measurement of soybean allergen emission levels at industrial sites. These methods may be used to monitor unloading and the impact of environmental interventions. PMID- 19123436 TI - T- helper 1 and 2 cytokine assay in patients with common variable immunodeficiency. AB - BACKGROUND: Common variable immunodeficiency (CVID) is a heterogeneous group of disorders, characterized by decreased immunoglobulin levels in serum and increased susceptibility to recurrent infections, autoimmunity, and malignancy. The pathogenesis of CVID is still unknown. OBJECTIVES: This study was performed to investigate T-helper (T(H)) 1 and 2 cytokine levels in patients with CVID. MATERIAL AND METHODS: Twenty-four cases of CVID were studied. Cytokine levels of interleukin (IL)-2, IL-4, IL-10, and interferon gamma were measured in the serum of the patients and compared with those of healthy controls. RESULTS: T(H)2 cytokine levels (IL-4 and IL-10) were significantly higher in the patient group than in the controls (median: 64.5 vs 0.0 pg/mL, P=.016 for IL-4 and 321.1 vs. 0.0 pg/mL, P=.001 for IL-10). However, there were no significant differences in T(H)1 cytokines between the 2 groups (median, 116.5 vs. 104.5 pg/mL, P=.22 for IL 2 and 50.5 vs. 42.3 pg/mL, P=.32 for IFN-gamma). CONCLUSIONS: Increased levels of IL-4 and IL-10 could indicate high activation of T(H)2 lymphocytes in this group of patients and consequently supports the concept of a bias towards T(H)2-type responses. PMID- 19123437 TI - Chitosan microparticles loaded with mite group 2 allergen Der f 2 alleviate asthma in mice. AB - BACKGROUND: In recent years, nanoparticle materials have found wide applications in drug transport and release systems. Chitosan is a good carrier for proteins, peptides, and nucleic acids because of its favorable release properties and ability to increase membrane permeability. OBJECTIVE: To investigate the effect of chitosan microparticles loaded with the major epitope peptide of mite group 2 allergen Der f 2 from Dermatophagoides farinae (Der f 2-47-67) in alleviating asthma in mice. METHODS: Derf 2-47-67 was entrapped in chitosan to obtain Derf 2 47-67-loaded chitosan microparticles, which were injected intraperitoneally into BALB/c mice prior to an intranasal challenge with a Der f extract allergen. Airway hyperreactivity was measured via whole-body plethysmography, and bronchoalveolar lavage (BAL) fluid was collected to calculate total cell and eosinophil counts. Changes in lung histology were assessed after hematoxylin eosin staining, and serum levels of Der f-specific immunoglobulin (Ig) G2a and IgE were determined by enzyme-linked immunoabsorbent assay. RESULTS: Mice immunized with Der f 2-47-67-loaded chitosan microparticles displayed decreased airway hyperreactivity, reduced numbers of eosinophils in BAL fluid, alleviated lung inflammation and mucus production, a reduced serum level of Der f-specific IgE and an increased serum level of Der f-specific IgG2a. CONCLUSION: Our data showed that Der f 2-47-67-loaded chitosan microparticles inhibited airway allergic inflammation. PMID- 19123438 TI - Acute generalized exanthematous pustulosis as a manifestation of carbamazepine hypersensitivity syndrome. AB - Anticonvulsant hypersensitivity syndrome (AHS) is a multisystemic disorder involving cutaneous changes and typical blood abnormalities that can be triggered by aromatic anticonvulsant drugs.The syndrome is commonly associated with a macular or papular rash or erythroderma. Acute generalized exanthematous pustulosis is a very rare cutaneous manifestation of AHS. A 41-year-old man was referred to our hospital for evaluation of a 3-day history of fever, leukocytosis, and generalized skin eruption. The patient had been taking carbamazepine for 1 month to treat hand tremor following surgery for intracerebral hemorrhage. Physical examination revealed facial edema and a large number of variable-sized pustules covering the body. Initial laboratory testing showed peripheral blood eosinophilia and abnormal liver function.A biopsy of pustular lesions revealed intraepidermal pustules, with perivascular lymphocytic infiltration. The skin lesions and laboratory results improved after withdrawal of carbamazepine and treatment with oral corticosteroids. PMID- 19123439 TI - Food-dependent exercise-induced anaphylaxis to lentil and anaphylaxis to chickpea in a 17-year-old boy. AB - We report the case of a 17-year-old boy who experienced 4 episodes of exercise induced anaphylactic reaction after ingestion of lentil and 2 episodes of anaphylaxis following ingestion of chickpea. His medical history revealed that he had allergic rhinitis with positive results after skin prick tests (SPT) with mites. His SPTs and specific immunoglobulin E antibody testing with lentil and chickpea were positive. Oral challenge with chickpea was not performed due to patient refusal. Treadmill exercise challenge tests in the fasting state and 1 hour after a meal not containing lentil were negative. However, an exercise challenge test 1 hour after intake of lentil soup resulted in pruritus of the hands, forearms, shoulders, and back, urticarial lesions on the face and shoulders, mild angioedema of the lips, and mild hoarseness and cough. To our knowledge, this is the first case of food-dependent exercise-induced anaphylaxis due to lentil. PMID- 19123440 TI - Necrosis of nasal cartilage due to mucormycosis in a patient with severe congenital neutropenia due to HAX1 deficiency. AB - Severe congenital neutropenia (SCN) is a primary immunodeficiency disease characterized by early onset of severe bacterial infection and persistent severe neutropenia. We describe an SCN patient with a history of recurrent infections. The clinical course was complicated by necrosis of the nasal cartilage due to mucormycosis. Molecular studies revealed a homozygous germline HAX1 mutation. Fungal infections may lead to serious complications in immunodeficient patients. Recurrent and severe infections should alert physicians to possible immunodeficiency disease. Early diagnosis and appropriate treatment are the most important keys to preventing irreversible organ damage. PMID- 19123441 TI - Diagnosis of clavulanic acid allergy using basophil activation and leukotriene release by basophils. AB - Clavulanic acid is a potent inhibitor of B-lactamase that is increasingly prescribed in association with amoxicillin. We report 2 cases of patients who experienced pruritus, wheals, and angioedema after oral intake of amoxicillin/clavulanic acid. Routine skin tests for B-lactam antibiotics and specific immunoglobulin (Ig) E were negative in both patients. Analysis of CD63 expression by the basophil activation test (BAT) using flow cytometry and of sulphidoleukotriene (sLT) release by basophils using the cellular allergen stimulation test (CAST) revealed significant positive responses with amoxicillin/clavulanic acid and with clavulanic acid, and negative responses with amoxicillin and other beta-lactam antibiotics. In addition, cultured CD3+CD4+ cells showed a significant increase in the expression of CD69, CD25, and HLA-DR in the presence of clavulanic acid. Both patients tolerated therapeutic doses of amoxicillin. BAT and CAST are useful ex vivo procedures for the detection of specific IgE-mediated allergy to clavulanic acid, especially for patients with negative skin test results. PMID- 19123442 TI - Identification of a 27 kDa protein in patients with anaphylactic reactions to mango. AB - Mango fruit has become increasingly popular in recent years. We report on 2 patients who developed anaphylactic reactions after the ingestion of fresh mango. Allergy to mango was confirmed by a positive skin prick test result and positive cellular allergen stimulation test results. Neither of the patients had detectable mango-specific immunoglobulin (Ig) E levels. Results were validated by sodium dodecyl sulfate polyacrylamide gel electrophoresis immunoblotting and analyzed using Quantiscan.We identified 2 major allergens with a molecular weight of 27 kDa in both patients, in addition to a 15 kDa allergen in 1 patient and a 32 kDa allergen in the other. Currently available IgE systems seem to be lacking these mango allergens and as such are probably unsuitable for diagnosing type 1 sensitization to mango. Skin prick testing with fresh mango fruit therefore seems to be a much more reliable test method for clinical practice. PMID- 19123443 TI - Collagenous colitis in a patient with common variable immunodeficiency. PMID- 19123444 TI - High risk of immediate-type reactions to soy drinks in 50 patients with birch pollinosis. PMID- 19123445 TI - House-dust endotoxin exposure and recurrent wheezing in infants: a cohort study. PMID- 19123446 TI - Selective immediate hypersensitivity to etoricoxib. PMID- 19123447 TI - Allergic rhinoconjunctivitis after ingestion of boiled rice. PMID- 19123448 TI - An unusual clinical presentation: invasive Candida non-albicans infections in ataxia telangiectasia. PMID- 19123449 TI - Effect of omalizumab treatment in a baker with occupational asthma. PMID- 19123450 TI - Pomegranate-dependent exercise-induced anaphylaxis. PMID- 19123451 TI - The basophil activation test in the diagnosis and follow-up of hymenoptera venom allergy: an alternative point of view. PMID- 19123452 TI - Nonasthmatic eosinophilic bronchitis in a baker caused by fungal alpha-amylase and wheat flour. PMID- 19123453 TI - The atypical Spitz tumor of uncertain biologic potential: a series of 67 patients from a single institution. AB - BACKGROUND: Atypical Spitz tumors (AST) are rare spitzoid melanocytic proliferations with an uncertain malignant potential. ASTs have overlapping features of both Spitz nevi and spitzoid melanoma, and consequently generate controversy with diagnosis and management. Sentinel lymph node biopsy (SLNB) has been proposed as a possible means to gain additional insight into the true biologic potential of these tumors; however, previous reports on the use of SLNB in ASTs have been limited by small numbers of patients and short durations of follow-up. METHODS: The authors extracted data from their institution's prospective melanoma database, collected between 1994 and 2007, for all patients with ASTs of uncertain biologic potential. They reviewed the clinical features of these patients, including the sentinel lymph node status, and the histological features of the tumors. RESULTS: A total of 67 patients with ASTs were identified, with a median age of 23.7 years. The mean depth was 2.4 mm. Of these, 57 had a SLNB performed, with 27 (47%) having a positive sentinel lymph node. SLNB-positive cases had a significantly lower mean age than SLNB-negative cases (17.9 vs 28.7 years; P = .013); however, no other significant differences were observed. All 27 patients with a positive SLNB were alive and disease free with median follow-up of 43.8 months. One patient who did not receive a SLNB developed recurrent disease with regional and distant metastases. CONCLUSIONS: ASTs do not appear to behave like conventional melanoma. There is a high incidence of microscopic lymph node deposits in SLNBs, but despite this finding, patients have a favorable prognosis. Our findings raise several questions regarding the malignant potential of ASTs, and the role of SLNB in their management. PMID- 19123455 TI - Temperament and character predictors of depressed mood over a 4-year interval. AB - BACKGROUND: The relationship between personality and depressed mood is not well understood. The present research sought to clarify the contributions of personality to an understanding of the development, course, and maintenance of depressed mood over time. METHODS: A large community sample [n=5591, 57% women, M age=551.01] was administered the Revised Temperament and Character Inventory [TCI R; Cloninger, 1999]: The Temperament and Character Inventory-Revised. St. Louis, MO: Center for Psychobiology of Personality, Washington University] and a modified version of the Center for Epidemiologic Studies-Depression Scale [CES-D; Radloff, 1977]: Appl Psychol Measure 1:385-401] at Time 1 [T1] and the Center for Epidemiologic Studies-Depression Scale again 4 years later at Time 2 [T2]. RESULTS: Whereas three of the seven personality dimensions were significantly and uniquely associated with depressed mood at T1, only harm avoidance emerged as a significant unique positive predictor of depressed mood at T2. When participants were grouped according to the level of stability versus change in depressed mood during the 4-year period, reward dependence, self-directedness , and cooperativeness were found to be significantly lower among those low in depressed mood at T1 who subsequently evidenced moderate to high levels of depressed mood at T2 when compared to those with low depressed mood on both assessment occasions. Additional analyses indicated that those who evidenced moderate to high levels of depression at T1 and T2 were distinctly higher on harm avoidance and lower on self-directedness when compared to those who had consistently low or variable levels of depressed mood across the two assessment occasions. CONCLUSIONS: Findings from this research are discussed in terms of personality related vulnerability and risk factors for future episodes of depressed mood. PMID- 19123454 TI - Exploration of anxiety sensitivity and distress tolerance as vulnerability factors for hoarding behaviors. AB - BACKGROUND: The phenomenon of compulsive hoarding, characterized by the acquisition of and failure to discard a large number of possessions, is increasingly recognized as a significant public health burden. Despite the magnitude of the impairment associated with this condition, empirical research is still in the nascent stages and many facets of the phenomenology, underlying vulnerability and risk factors for hoarding, are as of yet unknown. METHOD: The overall aim of the current investigation was to examine the association between hoarding behaviors and two potential vulnerability factors-anxiety sensitivity (AS) and distress tolerance (DT). In addition, we investigated the robustness of these associations as well as the interaction between the two hypothesized risk factors. Three studies (total N=745) involving independent nonclinical samples assessed hoarding, AS, DT, and relevant covariates using a range of measures. Resutlts: Findings revealed that AS and hoarding are significantly and robustly associated with one another beyond general depressive, anxiety, and nonhoarding obsessive-compulsive symptoms. Hoarding was also found to be associated with low DT. Consistent with prediction, AS and DT interacted such that DT may play a less important role among individuals with low AS. By contrast, low DT appears to increase vulnerability to hoarding symptoms among individuals high in AS. Results are discussed with regard to future research and treatment implications. Depression and Anxiety, 2009. Published 2009 Wiley-Liss, Inc. PMID- 19123456 TI - Treatment of major depressive disorder in the Finnish general population. AB - BACKGROUND: Few general population studies of the treatment of major depressive disorder (MDD) have included the whole spectrum of treatments. We estimated the rates of different treatments and the effect of individual and disorder characteristics plus provider type on treatment received. METHODS: In the Health 2000 Study, a representative sample (n=6,005) from the adult Finnish population (> or =30 years) were interviewed (CIDI) in 2000-2001 for the presence of DSM-IV mental disorders during the past 12 months. Logistic regression models were used to examine factors influencing the type of treatment: either pharmacotherapies (antidepressants, anxiolytics, sedatives/hypnotics, antipsychotics) or psychological treatment. RESULTS: Of the individuals with MDD (n=288), currently 24% used antidepressants, 11% anxiolytics, 16% sedatives/hypnotics, 5% antipsychotics, and 17% reported having received psychological treatment. Overall, 31% received antidepressants or psychological treatment or both; 18% received minimally adequate treatment. Of those 33% (n=94) using health care services for mental reasons, 76% received antidepressants or psychological treatment or both; 54% received minimal adequate treatment. In logistic regression models, the use of antidepressants was associated with female sex, being single, severe MDD, perceived disability, and comorbid dysthymic disorder; psychological treatment with being divorced, perceived disability, and comorbid anxiety disorder. CONCLUSIONS: Due to the low use of health services for mental reasons, only one-third of subjects with MDD use antidepressants, and less than one-fifth receives psychological treatment. The treatments provided are determined mostly by clinical factors such as severity and comorbidity, in part by sex and marital status, but not education or income. PMID- 19123457 TI - Natural remedies for anxiety disorders: potential use and clinical applications. AB - BACKGROUND: Natural remedies have been widely used and generally accepted as established treatments of depressive disorders, leading to the investigation of their potential role and efficacy in the treatment of the various anxiety disorders. METHODS: Numerous case reports, open-label, and placebo-controlled trials investigating the use of natural remedies in the treatment of anxiety disorders have yielded some encouraging results. RESULTS: Overall, these studies have indicated a potential role for natural remedies in the treatment of anxiety and suggest that such agents may possess a safer side effects profile when compared to conventional agents. However, these early findings, albeit promising, are yet to be supported by further investigation in large-scale, placebo controlled studies. CONCLUSIONS: This article reviews past and present research being performed in this area of clinical interest, while also revealing a remarkable paucity of data. PMID- 19123458 TI - Absolute lymphocyte count at the time of first relapse predicts survival in patients with diffuse large B-cell lymphoma. AB - Peripheral blood absolute lymphocyte count (ALC) is a survival prognostic factor in hematological malignancies. No reports have addressed whether ALC at the time of first relapse (ALC-R) predicts survival. Thus, we assessed the prognostic significance of ALC-R in diffuse large B-cell lymphoma (DLBCL). Patients were required to have been diagnosed with first relapsed DLBCL, have ALC-R values, and to be followed at Mayo Clinic, Rochester. From Feb 1987 until March 2006, 97 first relapsed DLBCL patients qualified for the study. The overall survival (OS) and progression-free survival (PFS) were measured from the time of first relapse. The value of ALC-R >or= 1.0 x 10(9)/L was used for the analysis. Both groups (ALC R >or= 1 or < 1 x 10(9)/L) were balanced for the international prognostic index at relapse (IPI-R) (P = 0.3), and for autologous stem cell transplantation (P = 0.4). Superior OS and PFS were observed with an ALC-R >or= 1.0 x 10(9)/L (N = 60) versus ALC-R < 1.0 x 10(9)/L (N = 37) [median OS: 28.7 months, 5 years OS rates of 39% versus median OS: 10.2 months, 5 years OS rates of 14%, P < 0.002; and median PFS: 14.8 months, 5 years PFS rates of 21% versus median PFS: 6.5 months, 5 years PFS rates of 8%, P < 0.004, respectively]. ALC-R was an independent prognostic factor for OS [RR = 0.4, P < 0.01] and PFS [RR = 0.5, P < 0.005]. ALC R predicts survival suggesting that host immunity is an important variable predicting survival in first relapsed DLBCL. PMID- 19123459 TI - Minimal residual disease monitoring after allogeneic transplantation may help to individualize post-transplant therapeutic strategies in acute myeloid malignancies. AB - This study evaluates the prognostic value of minimal residual disease (MRD) monitoring by multiparametric flow cytometry in 41 patients with acute myeloid leukemia or myelodysplastic syndrome undergoing allogeneic transplantation. MRD assessment after transplant (day +100) allowed to discriminate different risk populations, being the most significant cut-off value for outcome level of MRD < or > or = 10(-3). Outcome was significantly better among patients with low (<10( 3)) versus high (> or = 10(-3)) MRD at day +100 after transplant. Thus, overall survival was 73% versus 25% at 4 years among patients with low versus high MRD at day +100 after transplant (P = 0.002); 74% of patients with low MRD were event free at 4 years as compared to 17% among patients with high MRD (P = 0.01). In multivariate analysis, MRD value as well as chronic GVHD significantly influenced outcome. In conclusion, MRD monitoring early post-transplant is an important tool for outcome prediction and should be considered in decision making after allogeneic transplantation. PMID- 19123460 TI - Efficacy and safety of rituximab in adults' warm antibody autoimmune haemolytic anemia: retrospective analysis of 27 cases. AB - To better assess the efficacy and safety of rituximab in adults' warm antibody autoimmune hemolytic anemia (wAIHA), we conducted a retrospective study including 27 adults (mean age 49.7 +/- 21 years) with either primary (n = 17) or secondary (n = 10) wAIHA. On average, the patients received 2.1 +/- 1.4 treatment lines before rituximab and six had undergone splenectomy. Five patients were resistant to corticosteroids, 16 had a corticosteroid-dependent wAIHA and six had relapsed after an initial remission. Overall, 25/27 (93%) patients achieved an initial response from rituximab (eight complete responses and 17 partial responses). During a mean follow-up of 20.9 months after rituximab, five of the responders relapsed, three of whom were successfully retreated with rituximab. Two mild infusion-related-reactions occurred, one patient had a rituximab-related severe neutropenia and one case of pneumocystis jiroveci pneumonia occurred in a severely immunocompromized patient. In conclusion, rituximab seems highly effective and relatively safe in adults with steroid-resistant or steroid dependent wAIHA. PMID- 19123461 TI - Survival in young patients with intermediate- / high-risk myelofibrosis: estimates derived from databases for non transplant patients. AB - Recent studies have suggested that allogenic stem cell transplantation (allo-SCT) might be a better treatment option, compared to drug therapy, for young patients with high-/intermediate-risk primary myelofibrosis (PMF). However, there are no controlled studies that validate this contention and allo-SCT is associated with a substantial risk of procedure-related mortality and morbidity. In a retrospective analysis of nontransplant PMF patients, who were both young (age <60 years) and with high-/intermediate-risk disease, 1- and 3-year survival estimates were 87% and 55%, 95% and 77%, 71% and 58%, respectively, involving patients seen at three different centers with expertise in PMF; these data did not appear to be inferior to those published in the context of either myeloablative or reduced-intensity conditioning allo-SCT. These observations underscore the need for controlled studies to accurately assess the value of allo SCT in PMF. PMID- 19123462 TI - The methylenetetrahydrofolate reductase C677T mutation induces cell-specific changes in genomic DNA methylation and uracil misincorporation: a possible molecular basis for the site-specific cancer risk modification. AB - The C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with a decreased risk of colon cancer although it may increase the risk of breast cancer. This polymorphism is associated with changes in intracellular folate cofactors, which may affect DNA methylation and synthesis via altered one-carbon transfer reactions. We investigated the effect of this mutation on DNA methylation and uracil misincorporation and its interaction with exogenous folate in further modulating these biomarkers of one-carbon transfer reactions in an in vitro model of the MTHFR 677T mutation in HCT116 colon and MDA MB-435 breast adenocarcinoma cells. In HCT116 cells, the MTHFR 677T mutation was associated with significantly increased genomic DNA methylation when folate supply was adequate or high; however, in the setting of folate insufficiency, this mutation was associated with significantly decreased genomic DNA methylation. In contrast, in MDA-MB-435 cells, the MTHFR 677T mutation was associated with significantly decreased genomic DNA methylation when folate supply was adequate or high and with no effect when folate supply was low. The MTHFR 677T mutation was associated with a nonsignificant trend toward decreased and increased uracil misincorporation in HCT116 and MDA-MB-435 cells, respectively. Our data demonstrate for the first time a functional consequence of changes in intracellular folate cofactors resulting from the MTHFR 677T mutation in cells derived from the target organs of interest, thus providing a plausible cellular mechanism that may partly explain the site-specific modification of colon and breast cancer risks associated with the MTHFR C677T mutation. PMID- 19123464 TI - Diesel motor emissions and lung cancer mortality--results of the second follow-up of a cohort study in potash miners. AB - International health authorities have graded diesel motor emissions (DME) as probably cancerogenic in human beings. There are gaps in epidemiological evidence regarding exact exposure quantification, confounder control and the investigation of highly exposed populations. We investigated the association of DME and lung cancer mortality in a historical cohort study of 5,862 German potash miners who were followed from 1970 to 2001. Cumulative exposure (CE) was measured by representative concentrations of total carbon multiplied with exposure years from the mines' medical records. Exposure and smoking behavior were validated by interviews of 3,087 participants. We computed standardized mortality ratios (SMR, external comparison) and performed Cox regression (internal comparison). The relative risk estimates (RR) with 95%-confidence intervals were adjusted for age and smoking. Vital status and causes of death were confirmed for 98.1% of participants. Sixty-one lung cancer deaths occurred. SMR-analysis showed lower than expected lung cancer mortality (healthy-worker-effect). Internal comparisons revealed risk elevations from moderate to risk doubling depending on the exposure categories used (dichotomized: up to RR 1.43[0.67-3.03] for a CE of 4.90[mg/m(3)]*years as compared with less exposure; quintiles: RR 1.13[0.46 2.75], 2.47[1.02-6.02], 1.50[0.56-4.04] and 2.28[0.87-5.97] for a CE up to 2.04, 2.73, 3.90 and >3.90, respectively, as compared with the reference of <1.29[mg/m(3)]*years). Additional adjustment of length of follow-up leads to further RR increases and indicates healthy-worker-survivor-phenomena. The analyses of a sub-cohort (n = 3,335) with particularly accurate exposure measurement revealed a nonsignificant dose-response-relationship. Our results support an association of DME and lung cancer mortality. PMID- 19123463 TI - Physical activity, sedentary behavior, and endometrial cancer risk in the NIH AARP Diet and Health Study. AB - Consistent with a strong hormonal etiology, endometrial cancer is thought to be influenced by both obesity and physical activity. Although obesity has been consistently related to risk, associations with physical activity have been inconclusive. We examined relationships of activity patterns with endometrial cancer incidence in the NIH-AARP Diet and Health Study cohort, which included 109,621 women, ages 50-71, without cancer history, who in 1995-1996 completed a mailed baseline questionnaire capturing daily routine and vigorous (defined as any period of >or=20 min of activity at work or home causing increases in breathing, heart rate, or sweating) physical activity. A second questionnaire, completed by 70,351 women, in 1996-1997 collected additional physical activity information. State cancer registry linkage identified 1,052 primary incident endometrial cancers from baseline through December 31, 2003. In multivariate proportional hazards models, vigorous activity was inversely associated with endometrial cancer in a dose-response manner (p for trend = 0.02) (relative risk (RR) for >or=5 times/week vs. never/rarely = 0.77, 95% confidence interval (CI): 0.63-0.95); this association was more pronounced among overweight and obese women (body mass index >or=25; RR = 0.61, 95% CI: 0.47-0.79) than among lean women (body mass index <25; RR = 0.76, 95% CI: 0.52-1.10; p for interaction = 0.12). Although we observed no associations with light/moderate, daily routine or occupational physical activities, risk did increase with number of hours of daily sitting (p for trend = 0.02). Associations with vigorous activities, which may interact with body mass index, suggest directions for future research to clarify underlying biologic mechanisms, including those relating to hormonal alterations. PMID- 19123465 TI - Hypoxia-mediated up-regulation of MGr1-Ag/37LRP in gastric cancers occurs via hypoxia-inducible-factor 1-dependent mechanism and contributes to drug resistance. AB - Our previous study demonstrated hypoxia-inducible factor-1(HIF-1) could prompt multidrug resistance (MDR) phenotype and MGr1-Ag/37LRP, a novel drug-resistance protein was reported by our labortary, associated with multidrug resistance in gastric cancer. Given this association, we hypothesized that MGr1-Ag/37LRP contributed to HIF-1-dependent hypoxia-induced MDR phenotype. Initial experiments revealed that blocking MGr1-Ag/37LRP expression by siRNA in gastric cancer cells effectively reversed multidrug resistance phenotype induced by hypoxia. Subsequent analysis of MGr1-Ag/37LRP mRNA and protein in gastric cancer cells revealed a time-dependent manner increase with hypoxia. While the up-regulation of MGr1-Ag/37LRP was abolished by HIF-1 inhibition with siRNA. Studies using luciferase promoter constructs revealed a significant increase in activity in cells subject to hypoxia and such hypoxia inducibility was lost in cells co transfected siRNA targeting HIF-1. Analysis of the MGr1-Ag/37LRP promoter revealed several potential binding sites for HIF-1. Electrophoretic mobility shift assay and chromatin immunoprecipitation demonstrated a functional HIF-1 binding site within MGr1-Ag/37LRP gene regulatory sequence located at -16 to -11 relative to the transcriptional initiation point. These observations demonstrate that MGr1-Ag/37LRP is actively engaged by hypoxia and represent a novel HIF-1 target. Such results suggest hypoxia-elicited MGr1-Ag/37LRP expression as a pathway for resistance of gastric cancer to chemotherapeutics. PMID- 19123466 TI - Low-risk factor profile, estrogen levels, and breast cancer risk among postmenopausal women. AB - Obesity, alcohol consumption, physical inactivity and postmenopausal hormone use are known modifiable risk factors for breast cancer. We aim to measure incidence rates of breast cancer for women with favorable levels on all 4 risk factors (BMI or=60 years (OR = 1.35, 95% CI 1.09-1.66) and having skin that tanned easily (OR = 1.96, 95% CI 1.38-2.78). Participants from low-income households, and those who frequently used sun-protective clothing were more likely to have reduced sun protection practices (OR = 1.33, 95% CI 1.10-1.73 and OR = 1.73, 95% CI 1.36 2.20, respectively). This study provides evidence of reductions in sun protection practices in a population living in a high UV environment. There is an urgent need to refocus messages regarding sun exposure and for continued sun protection practices. PMID- 19123472 TI - Prospective cohort study of lifetime physical activity and breast cancer survival. AB - Few studies have examined the association between physical activity and disease outcomes in breast cancer survivors. Here, we report the association between lifetime total physical activity performed prior to diagnosis and disease outcomes in a population-based sample of breast cancer survivors. A cohort of 1,231 women diagnosed with breast cancer between 1995 and 1997 was followed for a minimum of 8.3 years for any cancer progressions, recurrences and new primaries; and a minimum of 10.3 years for deaths. All treatment and follow-up care received was abstracted from medical records. Data on physical activity including type (occupational, household, recreational) and dose (frequency, intensity and duration) performed during the entire lifetime until diagnosis were examined in Cox proportional hazards models as well as with cumulative incidence curves. An average of 126 MET-hr/week were reported for total physical activity (of which 13.9, 46.9 and 65.3 MET-hr/week were, respectively, for recreational, occupational and household activity). A decreased risk of breast cancer death and all deaths was observed among women in the highest versus the lowest quartiles of recreational activity (MET-hr/week/year) (HR = 0.54, 95% CI = 0.36-0.79). Both moderate (0.56, 95% CI = 0.38-0.82) and vigorous intensity recreational activity (0.74, 95% CI = 0.56-0.98) decreased the risk of breast cancer death. Moderate intensity recreational activity decreased the risk of a recurrence, progression or new primary cancer (0.66, 95% CI = 0.48-0.91). No other association with breast cancer survival was observed for other types of physical activity. Prediagnosis recreational activity conferred a benefit for survival after breast cancer. Moderate intensity recreational activity was particularly protective. PMID- 19123473 TI - The effect of cantharidins on leukemic stem cells. AB - To identify an agent with specific activity against leukemic stem cells (LSCs), we evaluated compounds that targeted hepatic leukemia factor (HLF), a gene implicated in hematopoietic stem cell (HSCs) regulation, that we found overexpressed in LSCs. Cantharidin, a natural toxin from blister beetles, used as medicinal agent since antiquity, has been described to modulate the HLF competitor NFIL3 and is under clinical evaluation as an antitumor and antimetastatic agent. The molecule is not a substrate for multidrug resistant pumps and does not cause myelosuppression, and therefore it represents a promising compound for selective ablation of LSCs. Cantharidin and norcantharidin, a derivative with reduced toxicity, decreased HLF protein levels and induced apoptosis in the AML cell line MV4-11 by modulating the expression of several molecules that govern survival pathway, including HLF, SLUG, NFIL3 and c myc, thereby inducing p53 and the mitochondrial caspase cascade. In vitro, cantharidin readily targeted primary AML stem and progenitor cells in contrast to conventional chemotherapeutic agents, such as Ara-C and daunorubicin, that mainly targeted more differentiated leukemic cells. In vitro the compound did not exhibit a therapeutic window, being equally toxic to normal HSCs and LSCs. In vivo cantharidin did not produce myelosuppression. Because of dose-limiting toxicity in vivo, neither cantharidin nor norcantharidin proved therapeutical benefit in AML xenograft models as a single agent. However, its potent in vitro LSC activity and pathway targeting may still be exploited clinically with a new generation of cantharidin derivatives or formulations and with appropriate drug combinations. PMID- 19123474 TI - Valproic acid enhances bosutinib cytotoxicity in colon cancer cells. AB - Unbalanced histone deacetylase (HDAC) hyperactivity is a common feature of tumor cells. Inhibition of HDAC activity is often associated with cancer cell growth impairment and death. Valproic acid (VPA) is a HDAC inhibitor used for the treatment of epilepsy. It has recently been recognized as a promising anticancer drug. We investigated the effects of VPA on growth and survival of colon cancer cells. VPA caused growth inhibition and programmed cell death that correlated with histone hyperacetylation. VPA modulated the expression of various factors involved in cell cycle control and apoptosis and induced caspase activation. Interestingly, VPA induced downregulation of c-Src and potentiated the cytotoxic effects of the c-Src inhibitor bosutinib, both in vitro and in vivo. The combination of sublethal doses of VPA and bosutinib led to massive apoptosis of colon cancer cells, irrespective of their genetic background. These results suggest that VPA may be employed as a positive modulator of bosutinib antitumor activity in colorectal cancer. PMID- 19123476 TI - DNA adduct formation and induction of micronuclei and mutations in B6C3F1/Tk mice treated neonatally with acrylamide or glycidamide. AB - Acrylamide, a food contaminant, is carcinogenic in experimental animals, with both genotoxic and nongenotoxic pathways being proposed. To obtain information regarding mechanisms of acrylamide tumorigenesis, we compared the extent of DNA adduct formation and induction of micronuclei and mutations in mice treated neonatally with acrylamide and its electrophilic metabolite glycidamide. Male and female B6C3F1/Tk mice were treated intraperitoneally on postnatal days (PNDs) 1, 8 and 15 or PNDs 1-8 with 0.14 or 0.70 mmol acrylamide or glycidamide per kg body weight per day. One day after the final dose, B6C3F1/Tk(+/+) mice were killed to measure DNA adduct levels and peripheral blood micronuclei. Three weeks after the last treatment, B6C3F1/Tk(+/-) mice were killed to assess the Hprt and Tk mutant frequencies in spleen lymphocytes. The levels of N7-(2-carbamoyl-2 hydroxyethyl)guanine, the major glycidamide-DNA adduct, decreased in the order 0.70 mmol glycidamide > 0.70 mmol acrylamide > 0.14 mmol glycidamide approximately 0.14 mmol acrylamide. Only glycidamide increased the frequency of micronucleated reticulocytes and normochromatic erythrocytes. In mice treated on PNDs 1, 8 and 15, the Hprt mutant frequency was increased by 0.70 mmol glycidamide. In mice dosed on PNDs 1-8, 0.70 mmol glycidamide caused extensive mortality; each of the other treatments increased the Tk mutant frequency, whereas acrylamide increased the Hprt mutant frequency. These data suggest that the mutagenic response in neonatal mice treated on PNDs 1, 8 and 15 is due to glycidamide, whereas mutations resulting from dosing on PNDs 1-8 are due to another mechanism. PMID- 19123477 TI - Two distinct expression patterns of urokinase, urokinase receptor and plasminogen activator inhibitor-1 in colon cancer liver metastases. AB - Metastatic growth and invasion by colon cancer cells in the liver requires the ability of the cancer cells to interact with the new tissue environment. Plasmin(ogen) is activated on cell surfaces by urokinase-type PA (uPA), and is regulated by uPAR and plasminogen activator inhibitor-1 (PAI-1). To compare the expression patterns of uPA, uPAR and PAI-1 in colon cancer with that in their liver metastases, we analysed matched samples from 14 patients. In all 14 primary colon cancers, we found upregulation of uPAR, uPA mRNA and PAI-1 in primarily stromal cells at the invasive front. In 5 of the 14 liver metastases, we found intense expression of uPAR, uPA-mRNA and PAI-1 in primarily stromal cells at the metastases periphery, and in an expression pattern similar to that found in the primary tumours. In the remaining 9 liver metastases, uPAR and uPA-mRNA were only seen associated with the presence of necrosis within the liver metastases. In addition, PAI-1-immunoreactivity was in all liver metastases seen in hepatocytes at the metastases periphery. Interestingly, the former 5 liver metastases positive for uPAR, uPA mRNA and PAI-1 at the metastasis periphery all had a predominantly desmoplastic reaction, whereas 8 of the remaining 9 showed direct contact between the cancer cells and the liver parenchyma. We conclude that there are 2 distinct patterns of expression of uPAR, uPA and PAI-1 in colon cancer liver metastases and that these correlate closely with 2 morphological growth patterns. These findings may have implication for the treatment of patients with metastatic disease. PMID- 19123475 TI - Salvianolic acid B inhibits growth of head and neck squamous cell carcinoma in vitro and in vivo via cyclooxygenase-2 and apoptotic pathways. AB - Overexpression of cyclooxygenase-2 (COX-2) in oral mucosa has been associated with increased risk of head and neck squamous cell carcinoma (HNSCC). Celecoxib is a nonsteroidal anti-inflammatory drug, which inhibits COX-2 but not COX-1. This selective COX-2 inhibitor holds promise as a cancer preventive agent. Concerns about cardiotoxicity of celecoxib, limits its use in long-term chemoprevention and therapy. Salvianolic acid B (Sal-B) is a leading bioactive component of Salvia miltiorrhiza Bge, which is used for treating neoplastic and chronic inflammatory diseases in China. The purpose of this study was to investigate the mechanisms by which Sal-B inhibits HNSCC growth. Sal-B was isolated from S. miltiorrhiza Bge by solvent extraction followed by 2 chromatographic steps. Pharmacological activity of Sal-B was assessed in HNSCC and other cell lines by estimating COX-2 expression, cell viability and caspase dependent apoptosis. Sal-B inhibited growth of HNSCC JHU-022 and JHU-013 cells with IC(50) of 18 and 50 microM, respectively. Nude mice with HNSCC solid tumor xenografts were treated with Sal-B (80 mg/kg/day) or celecoxib (5 mg/kg/day) for 25 days to investigate in vivo effects of the COX-2 inhibitors. Tumor volumes in Sal-B treated group were significantly lower than those in celecoxib treated or untreated control groups (p < 0.05). Sal-B inhibited COX-2 expression in cultured HNSCC cells and in HNSCC cells isolated from tumor xenografts. Sal-B also caused dose-dependent inhibition of prostaglandin E(2) synthesis, either with or without lipopolysaccharide stimulation. Taken together, Sal-B shows promise as a COX-2 targeted anticancer agent for HNSCC prevention and treatment. PMID- 19123478 TI - Hepatocarcinogenesis in mice with a conditional knockout of the hepatocyte growth factor receptor c-Met. AB - The receptor for the hepatocyte growth factor/scatter factor (HGF/SF), c-Met, plays a role in tumour promotion, progression and metastasis. In this study, we analysed chemically induced hepatocarcinogenesis in mice lacking a functional HGF receptor in their liver. Control and c-Met deficient mice were injected with a single dose of N-nitrosodiethylamine (DEN, 90 mICROg/g b.wt.) at 6 weeks of age and mice were subsequently kept on a phenobarbital (PB) containing diet (0.05%) for 35 weeks or on control diet. At the end of the experiment, the carcinogenic response in liver of the animals was monitored. Conditional c-met knockout (KO) mice showed a higher prevalence of macroscopically visible liver tumours and of glutamine synthetase positive and glucose-6-phosphatase deficient lesions in liver. Tumour promotion by PB led to significant increases in the number of preneoplastic and neoplastic lesions in liver of both wild-type and c-met knockout mice, with only minor differences in response. Our results indicate that a defect in c-Met-mediated signaling increases chemically induced tumour initiation in liver but does not significantly affect PB-mediated tumour promotion. PMID- 19123480 TI - Identification of a rhabdomyosarcoma targeting peptide by phage display with sequence similarities to the tumour lymphatic-homing peptide LyP-1. AB - Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children. To improve existing therapies and broaden the spectrum of cytotoxic agents that can be used in RMS treatment, we performed a phage-display-based screening for peptides that bind specifically to RMS cells. Two peptides binding to RMS and to other tumour cell lines, but not to normal skeletal muscle cells and fibroblasts, were isolated from phage-displayed random peptide libraries. One peptide, named RMS-I (CQQSNRGDRKRC) contained the integrin-binding motif RGD and its binding was blocked by an antibody against alpha(v)beta(3)integrin, which is expressed on the RMS cell line RD. The isolation of RMS-I confirmed the validity of our screening procedure. The second peptide, named RMS-II (CMGNKRSAKRPC), shows sequence similarity to a previously identified peptide with tumour lymphatic specificity, LyP-1. However, RMS-II binds in vivo to RMS xenografts better than LyP-1 and homes to the tumour blood and not to lymphatic vessels. Therefore, RMS-II represents a promising peptide for the development of RMS-specific targeting approaches. PMID- 19123479 TI - Pharmacogenomics and analogues of the antitumour agent N6-isopentenyladenosine. AB - N(6)-isopentenyladenosine (i(6)A), a member of the cytokinin family of plant hormones, has potent in vitro antitumour activity in different types of human epithelial cancer cell lines. Gene expression profile analysis of i(6)A-treated cells revealed induction of genes (e.g., PPP1R15A, DNAJB9, DDIT3, and HBP1) involved in the negative regulation of cell cycle progression and reportedly up regulated during cell cycle arrest in stress conditions. Of 6 i(6)A analogues synthesized, only the 1 with a saturated double bond of the isopentenyl side chain had in vitro antitumour activity, although weaker than that of i(6)A, suggesting that i(6)A biological activity is highly linked to its structure. In vivo analysis of i(6)A and the active analogue revealed no significant inhibition of cancer cell growth in mice by either reagent. Thus, although i(6)A may inhibit cell proliferation by regulating the cell cycle, further studies are needed to identify active analogues potentially useful in vivo. PMID- 19123481 TI - Three genetic developmental stages of papillary renal cell tumors: duplication of chromosome 1q marks fatal progression. AB - Papillary renal cell tumors (RCT) make up a cytomorphologically and biologically heterogeneous group of kidney cancers including renal cell adenomas (RCA) and renal cell carcinomas (RCC). To find genetic markers landmarking the tumor progression, we have evaluated the genetic alterations obtained by karyotyping, chromosomal and array-CGH and compared with the cytological characteristics and biological behavior of 60 papillary RCTs. Based on the genetic and clinical data, we have separated 3 groups of tumors and proposed 3 genetically defined developmental stages of papillary RCTs. Papillary RCAs are characterized by combined trisomy of chromosomes 7 and 17, whereas papillary RCCs displayed additional trisomies of 3q, 8q, 12q, 16q and 20q. In addition to the genetic changes occurring in the second group, the third group of tumors was characterized by 1q gain and 6q, 8p, 9p and 14q losses. Kaplan-Meier analysis revealed a significant association between chromosome 1q gain and deadly outcome of the disease. The cytomorphological variation and size of tumors in the second and third groups did not correlate with the clinical outcome. Therefore, we suggest that our genetic classification system landmarking papillary RCA, papillary RCC without and with progression offer a better system to characterize the tumor biology of clinical significance than a cellular/morphological classification. PMID- 19123482 TI - Elevated circulating adiponectin and elevated insulin sensitivity in adiponectin transgenic mice are not associated with reduced susceptibility to colon carcinogenesis. AB - Obesity, particularly visceral adiposity, is an established risk factor for colorectal cancer (CRC) and this is thought to result, at least in part, from insulin resistance and chronic hyperinsulinemia that may be mediated by adipokines. Serum levels of adiponectin, the most abundant protein secreted from adipocytes, are decreased in obesity and are inversely associated with insulin resistance and hyperinsulinemia. The objective of this study was to determine whether elevated circulating adiponectin plays a role in colon carcinogenesis using adiponectin transgenic (AdTg) mice that have 2-3-fold elevated circulating adiponectin but similar body weights as wildtype (WT) littermates used as controls. Eight-week old male and female AdTg and WT mice were treated with 4 weekly injections of the colon-specific carcinogen azoxymethane (AOM). One week following the last dose of AOM, all mice were placed on a high-fat diet and killed 24 weeks later, at 36 weeks of age, for the analysis of colon tumors. Glucose tolerance tests (GTT) were performed by injecting 2g/kg dextrose or 1.25 1.5 g/kg dextrose into all 12-week and 32-35-week-old mice respectively, and measuring blood from the tail vein 15, 30, 60 and 120 min following glucose administration. There were no significant differences in colon tumor incidence, number or size between AdTg and WT mice of either sex. AdTg mice of both sexes displayed resistance to diet-induced decreases in insulin sensitivity. Our results show that constitutively elevated levels of circulating adiponectin in AdTg mice do not confer protection against the development of colon tumors. PMID- 19123484 TI - Social anxiety in adolescents: the effect of video feedback on anxiety and the self-evaluation of performance. AB - BACKGROUND: A negative self-generated image is thought to play a role in the development and maintenance of social anxiety. Video feedback (VF) is an effective therapeutic technique for correcting this distorted image with adults during stressful social tasks. This study investigated the effectiveness of VF with adolescents. METHOD: Thirty-six highly socially anxious adolescents (13 to 17-year-olds) were randomly assigned to one of two groups: VF or no VF. Both groups gave two brief speeches to a video camera. Self-rated measures of anticipatory anxiety, predicted performance and actual performance during the speeches were completed at several time points. The speeches were also rated by two independent observers. RESULTS: Adolescents who received VF, in comparison with control participants, developed more positive appraisals of their performance during the first speech. Predictions of their performance on the second speech also improved, and a reduction in state anxiety was observed. Furthermore, a more positive appraisal of performance was generalized to the second speech. The independent observers could not distinguish between the participants who received VF and those who did not. CONCLUSIONS: When used with socially anxious adolescents, VF may be a beneficial therapeutic technique in correcting a distorted self-perception of performance, reducing anxiety prior to entering a stressful social task and enhancing predictions of performance for future tasks. PMID- 19123483 TI - In vitro and in vivo study of phloretin-induced apoptosis in human liver cancer cells involving inhibition of type II glucose transporter. AB - Phloretin (Ph), a natural product found in apples and pears with glucose transporter (GLUT) inhibitory activity, exerts antitumor effects. However, little is known about its effects on human liver cancer. The purpose of this study is to test the cytotoxic effects of Ph on HepG2 cells and to identify the underlying molecular pathways. Human hepatocellular carcinoma specimens and HepG2 show a high level of GLUT2 transporter activity in the cell membrane. Real-time PCR and MTT assays demonstrate that Ph-induced cytotoxicity correlates with the expression of GLUT2. Flow cytometry and DNA fragmentation studies show that 200 microM Ph induces apoptosis in HepG2, which was reversed by glucose pretreatment. GLUT2 siRNA knockdown induced HepG2 apoptosis, which was not reversed by glucose. Western blot analysis demonstrates that both intrinsic and extrinsic apoptotic pathways in addition to Akt and Bcl-2 family signaling pathways are involved in Ph-induced cell death in HepG2 cells. Furthermore, using flow cytometry analysis, a mitochondrial membrane potential assay and Western blot analysis, we show that cytochalasin B, a glucose transport inhibitor, enhances the Ph-induced apoptotic effect on HepG2 cells, which was reversed by pretreatment with glucose. Furthermore, we found significant antitumor effects in vivo by administering Ph at 10 mg/kg intraperitoneally to severe combined immune deficiency mice carrying a HepG2 xenograft. A microPET study in the HepG2 tumor-bearing mice showed a 10 fold decrease in (18)F-FDG uptake in Ph-treated tumors compared to controls. Taken together, these results suggest that Ph-induced apoptosis in HepG2 cells involves inhibition of GLUT2 glucose transport mechanisms. PMID- 19123487 TI - Anomalous melting behavior under extreme conditions: hard matter turning "soft". AB - We show that a system of particles interacting through the exp-6 pair potential, commonly used to describe effective interatomic forces under high compression, exhibits anomalous melting features such as reentrant melting and a rich solid polymorphism, including a stable BC8 crystal. We relate this behavior to the crossover, with increasing pressure, between two different regimes of local order that are associated with the two repulsive length scales of the potential. Our results provide a unifying picture for the high-pressure melting anomalies observed in many elements and point out that, under extreme conditions, atomic systems may reveal surprising similarities with soft matter. PMID- 19123488 TI - Charge on a weak polyelectrolyte. AB - Fluorescence measurements with single-molecule sensitivity are used to measure the hydrodynamic size and local pH of a weak polyelectrolyte, poly-2-vinyl pyridine end labeled with pH-sensitive dye, the polyelectrolyte having concentration so low (nanomolars) that molecular properties are resolvable only from fluorescence experiments and cannot be accessed by light scattering. We find that the local pH near the dye, inferred from its brightness, is consistently three orders of magnitude higher than the bulk pH. Upon varying the bulk pH, we measure the collapse point at which hydrophobic attraction overwhelms electrostatic repulsion between charged elements along the chain, and conclude that adding monovalent salt shifts this coil-to-globule collapse to higher pH than in the absence of salt. The influence of salt appears to shift the ionization equilibrium of this weak polyelectrolyte in the direction of the chain possessing enhanced electric charge at a given pH. Phenomenologically, this is opposite to the case for strong polyelectrolytes, although the mechanism differs. PMID- 19123489 TI - Constructing diabatic states from adiabatic states: extending generalized Mulliken-Hush to multiple charge centers with boys localization. AB - This article shows that, although Boys localization is usually applied to single electron orbitals, the Boys method itself can be applied to many electron molecular states. For the two-state charge-transfer problem, we show analytically that Boys localization yields the same charge-localized diabatic states as those found by generalized Mulliken-Hush theory. We suggest that for future work in electron transfer, where systems have more than two charge centers, one may benefit by using a variant of Boys localization to construct diabatic potential energy surfaces and extract electronic coupling matrix elements. We discuss two chemical examples of Boys localization and propose a generalization of the Boys algorithm for creating diabatic states with localized spin density that should be useful for Dexter triplet-triplet energy transfer. PMID- 19123490 TI - Ab initio calculation of interatomic decay rates of excited doubly ionized states in clusters. AB - Recently, a computational technique for ab initio calculation of the interatomic and intermolecular nonradiative decay processes has been developed [V. Averbukh and L. S. Cederbaum, J. Chem. Phys. 123, 204107 (2005)]. It combines the Fano formalism with the Green's function method known as the algebraic diagrammatic construction. The problem of normalization of continuum wave functions stemming from the use of the Gaussian basis sets is solved by using the Stieltjes imaging technique. In the present paper, the methodology is extended in order to describe the interatomic decay of excited doubly ionized states of clusters. The new computational scheme is applied to compute the interatomic decay rates of doubly ionized states formed by Auger relaxation of core vacancies in NeAr and MgNe van der Waals clusters. PMID- 19123491 TI - Structure and reactions of carbon and hydrogen on Ru(0001): a scanning tunneling microscopy study. AB - The interaction between carbon and hydrogen atoms on a Ru(0001) surface was studied using scanning tunneling microscopy (STM), density functional theory (DFT) and STM image calculations. Formation of CH species by reaction between adsorbed H and C was observed to occur readily at 100 K. When the coverage of H increased new complexes of the form of CH+nH (n=1, 2, and 3) were observed. These complexes, never observed before, might be precursors for further hydrogenation reactions. DFT analysis reveals that a considerable energy barrier exists for the CH+H-->CH(2) reaction. PMID- 19123492 TI - Variational and perturbative formulations of quantum mechanical/molecular mechanical free energy with mean-field embedding and its analytical gradients. AB - Conventional quantum chemical solvation theories are based on the mean-field embedding approximation. That is, the electronic wavefunction is calculated in the presence of the mean field of the environment. In this paper a direct quantum mechanical/molecular mechanical (QM/MM) analog of such a mean-field theory is formulated based on variational and perturbative frameworks. In the variational framework, an appropriate QM/MM free energy functional is defined and is minimized in terms of the trial wavefunction that best approximates the true QM wavefunction in a statistically averaged sense. Analytical free energy gradient is obtained, which takes the form of the gradient of effective QM energy calculated in the averaged MM potential. In the perturbative framework, the above variational procedure is shown to be equivalent to the first-order expansion of the QM energy (in the exact free energy expression) about the self-consistent reference field. This helps understand the relation between the variational procedure and the exact QM/MM free energy as well as existing QM/MM theories. Based on this, several ways are discussed for evaluating non-mean-field effects (i.e., statistical fluctuations of the QM wavefunction) that are neglected in the mean-field calculation. As an illustration, the method is applied to an S(N)2 Menshutkin reaction in water, NH(3)+CH(3)Cl-->NH(3)CH(3) (+)+Cl(-), for which free energy profiles are obtained at the Hartree-Fock, MP2, B3LYP, and BHHLYP levels by integrating the free energy gradient. Non-mean-field effects are evaluated to be <0.5 kcal/mol using a Gaussian fluctuation model for the environment, which suggests that those effects are rather small for the present reaction in water. PMID- 19123493 TI - Implementation and performance of a domain-specific basis set incremental approach for correlation energies: applications to hydrocarbons and a glycine oligomer. AB - The fully automated implementation of the incremental scheme for CCSD energies has been modified for the usage of a domain-specific basis set. We find that the computational effort can be reduced significantly without loss of accuracy. It is shown explicitly in applications on hydrocarbons and the (glycine)(4) oligomer that the error of the incremental expansion for the total energy is usually below 1 kcal/mol at third order. Furthermore, it is demonstrated that the proposed approach saves CPU time, random access memory, and disk space. Moreover, we show in various tests that the inherently parallel incremental calculations can be run on up to 50 CPUs without significant loss of computer time. PMID- 19123494 TI - Quantum Monte Carlo formulation of volume polarization in dielectric continuum theory. AB - We present a novel formulation based on quantum Monte Carlo techniques for the treatment of volume polarization due to quantum mechanical penetration of the solute charge density in the solvent domain. The method allows to accurately solve Poisson's equation of the solvation model coupled with the Schrodinger equation for the solute. We demonstrate the performance of the approach on a representative set of solutes in water solvent and give a detailed analysis of the dependence of the volume polarization on the solute cavity and the treatment of electron correlation. PMID- 19123495 TI - A note on the Pulay force at finite electronic temperatures. AB - Pulay's original expression for the basis-set dependent adjustment term to the Hellmann-Feynman force in electronic structure theory, which occurs for nonorthogonal local basis-set representations, is based on the idempotency condition of a pure ensemble. At finite electronic temperatures with a fractional occupation of the states, the conventional expression of the Pulay force is therefore no longer valid. Here we derive a simple and computationally efficient expression for a generalized Pulay force, which is suitable for large-scale ab initio simulations at finite electronic temperatures using local nonorthogonal basis-set representations. The generalized Pulay force expression is given in terms of the temperature-dependent density matrix. For the construction of the density matrix, we propose a recursive Fermi operator expansion algorithm that automatically converges to the correct chemical potential. PMID- 19123496 TI - Application of improved virtual orbital based multireference methods to N2, LiF, and C4H6 systems. AB - The improved virtual orbital (IVO) complete active space configuration interaction (CASCI) based multiconfigurational quasidegenerate perturbation theory (MCQDPT) and its single-root version (termed as MRMPPT) are applied to assess the efficacy and the reliability of these two methods. Applications involve the ground and/or excited state potential energy curves (PECs) of N(2), LiF, and C(4)H(6) (butadiene) molecules, systems that are sufficiently complex to assess the applicability of these methods. The ionic-neutral curve crossing involving the lowest two (1)Sigma(+) states of LiF molecule is studied using the IVO-MCQDPT method, while its single-root version (IVO-MRMPPT) is employed to study the ground state PEC for isomerization of butadiene and to model the bond dissociation of N(2) molecule. Comparisons with the standard methods (full CI, coupled cluster with singles and doubles, etc.) demonstrate that the IVO-based MRMPPT and MCQDPT approaches provide smooth and reliable PECs for all the systems studied. The IVO-CASCI method is explored to enable geometry optimization for ground state of C(4)H(6) using numerical energy gradients. The ground spectroscopic constants of N(2) and LiF determined using the numerical gradient based IVO-CASCI method are in accord with experiment and with other correlated calculations. As an illustration, we may point out that the maximum deviation from the experiment in our estimated normal mode frequency of LiF is 34 cm(-1), whereas for the bond length, the maximum error is just 0.012 A. PMID- 19123497 TI - Variational, V-representable, and variable-occupation-number perturbation theories. AB - Density-functional perturbation theory with variationally fitted Kohn-Sham (KS) potentials is described. Requiring the Fock matrix and density matrix to commute through each order of perturbation theory determines the off-diagonal elements of the density matrix, and thus the effect of changing occupation numbers in density functional perturbation theory. At each order of perturbation theory, the change in occupation numbers at that order enters only the diagonal part of the density matrix. The theory contains no phases, and a limiting process relates the rest of the diagonal density matrix element, obtained from wave function perturbation theory, to the off-diagonal part, obtained by commutation. V-representable density-functional theory is most practical when the KS potential is expanded in a finite basis to create the Sambe-Felton (SF) potential of analytic density functional theory. This reduces the dimensionality of perturbation theory from order N(2) in the orbital basis to order N in the SF basis. Computing the (occupied-virtual)(2), i.e., N(4), sum over states once at the end of a self consistent-field molecular orbital calculation removes the orbitals from all higher orders of perturbation theory. The rank-N(2) iterative coupled-perturbed equations are replaced by rank-N matrix inversion, to fit variationally the perturbed SF potential at each order. As an example of the 2n+1 rule of perturbation theory, the variational, first-order potential is used to give precise second and third derivatives of the energy with respect to occupation number. The hardness and hyperhardness are computed for a standard set of molecules. Both are essentially independent of how the variational SF potential is constrained for four different constraint combinations. With variational fitting, the precision of derivatives and the fidelity of the fit to the SF potential are not related. Analytic derivatives are accurate to machine precision for any constraint and all fitting basis sets. PMID- 19123498 TI - Calculation of first and second static hyperpolarizabilities of one- to three dimensional periodic compounds. Implementation in the CRYSTAL code. AB - A computational scheme for the evaluation of the static first (beta) and second (gamma) hyperpolarizability tensors of systems periodic in 1D (polymers), 2D (slabs), 3D (crystals), and, as a limiting case, 0D (molecules) has been implemented, within the coupled perturbed Hartree-Fock framework (CPHF), in the CRYSTAL code, which uses a Gaussian type basis set. This generalizes to 2D and 3D the work by Bishop et al. (J. Chem. Phys. 114, 7633 (2001)). CPHF is applied for beta and gamma (the polarizability tensor alpha is also reported for completeness) of LiF in different aggregation states: finite and infinite chains, slabs, and cubic crystal. Correctness of the computational scheme and its numerical efficiency are documented by the trend of beta and gamma for increasing dimensionality: for a finite linear chain containing N LiF units, the hyperpolarizability tends to the infinite chain value at large N, N parallel chains give the slab value when N is sufficiently large, and N superimposed slabs tend to the bulk value. High numerical accuracy can be achieved at relatively low cost, with a dependence on the computational parameters similar to that observed for field-free self-consistent field (SCF) calculations. PMID- 19123499 TI - Spin-free intermediate Hamiltonian Fock space coupled-cluster theory with full inclusion of triple excitations for restricted Hartree Fock based triplet states. AB - The recently reported inclusion of the connected triples into the intermediate Hamiltonian realization of the Fock space coupled-cluster (IH-FS-CC) theory [M. Musial and R. J. Bartlett, J. Chem. Phys. 129, 044101 (2008)] is extended to produce the triplet states. This is done entirely in spatial orbitals on the basis of the double occupancy in the restricted Hartree Fock reference function. New equations for the triplet state amplitudes expressed in terms of the Goldstone diagrams are derived and implemented. Several applications show the usefulness of the IH-FS-CC scheme to describe the triplet states with the computational gains inherent to a spin-free implementation. PMID- 19123501 TI - Contribution of the Gouy phase to two-pathway coherent control of the photoionization and photodissociation of vinyl chloride. AB - The electric field of a light wave accumulates a pi phase shift as it passes through a focus. We show here how this effect, known as the Gouy phase, may be used to control the branching ratio of a unimolecular reaction when the products are formed with different numbers of photons. We demonstrate this control method for the ionization and dissociation of vinyl chloride, using absorption of 177 and 532 nm photons to induce a pair of interfering paths. Excellent agreement between the observed and calculated phase shift as a function of the axial coordinate of the laser focus indicates that fragmentation occurs via a ladder switching mechanism. The axial dependence of the modulation depth is evidence of loss of coherence at higher internal temperatures of the molecule. PMID- 19123500 TI - Enhanced identification and exploitation of time scales for model reduction in stochastic chemical kinetics. AB - Widely different time scales are common in systems of chemical reactions and can be exploited to obtain reduced models applicable to the time scales of interest. These reduced models enable more efficient computation and simplify analysis. A classic example is the irreversible enzymatic reaction, for which separation of time scales in a deterministic mass action kinetics model results in approximate rate laws for the slow dynamics, such as that of Michaelis-Menten. Recently, several methods have been developed for separation of slow and fast time scales in chemical master equation (CME) descriptions of stochastic chemical kinetics, yielding separate reduced CMEs for the slow variables and the fast variables. The paper begins by systematizing the preliminary step of identifying slow and fast variables in a chemical system from a specification of the slow and fast reactions in the system. The authors then present an enhanced time-scale separation method that can extend the validity and improve the accuracy of existing methods by better accounting for slow reactions when equilibrating the fast subsystem. The resulting method is particularly accurate in systems such as enzymatic and protein interaction networks, where the rates of the slow reactions that modify the slow variables are not a function of the slow variables. The authors apply their methodology to the case of an irreversible enzymatic reaction and show that the resulting improvements in accuracy and validity are analogous to those obtained in the deterministic case by using the total quasi-steady-state approximation rather than the classical Michaelis-Menten. The other main contribution of this paper is to show how mass fluctuation kinetics models, which give approximate evolution equations for the means, variances, and covariances of the concentrations in a chemical system, can feed into time-scale-separation methods at a variety of stages. PMID- 19123502 TI - Gas-phase infrared multiple photon dissociation spectroscopy of isolated SF6- and SF5- anions. AB - Resonantly enhanced multiple photon dissociation of gas-phase SF(6) (-) and SF(5) (-) is studied using tunable infrared light from the FELIX free electron laser. The photodissociation spectrum of the sulfur hexafluoride anion, producing SF(5) (-), is recorded over the spectral range of 250-1650 cm(-1). The infrared multiple photon dissociation cross section exhibits a strong, broad resonance enhancement at 675 cm(-1) in agreement with the calculated value of nu(3), one of the two IR-active fundamental vibrational modes predicted for the O(h)-symmetry ion. Much weaker absorption features are observed in the spectral region of 300 450 cm(-1) as well as at 580 cm(-1) that are not easily assigned to the other IR active fundamental of SF(6) (-) since these resonances are observed at a much higher energy than the calculated values for the IR-active nu(4) mode. The potential role of binary combination bands is considered. Photodissociation from the sulfur pentafluoride anion produced only F(-), but photodetachment was also observed through SF(6) associative electron capture. The IR multiple photon dissociation spectrum of SF(5) (-) shows multiple resonances within the region of 400-900 cm(-1) and agreement with calculations is clear, including the observation of three fundamental frequencies: nu(1) at 780 cm(-1), nu(7) at 595 cm(-1), and nu(8) at 450 cm(-1). Comparisons of the measured frequencies with ab initio and density functional theory calculations confirm an SF(5) (-) anion of C(4v) symmetry. Similar comparisons for SF(6) (-) are not inconsistent with an anion of O(h) symmetry. PMID- 19123503 TI - Interaction of NO(A2Sigma+) with rare gas atoms: potential energy surfaces and spectroscopy. AB - We present the results of an ab initio study of the interaction of electronically excited NO(A (2)Sigma(+)) with rare gas (Rg) atoms. The bound states of each NO(A)-Rg species are determined from potential energy surfaces calculated at the RCCSD(T) level of theory. Making use of the NO(X (2)Pi)-Rg vibrational wavefunctions, we then simulate electronic spectra. For NO-Kr and NO-Xe we obtain good qualitative agreement with the previously published experimental spectra. For NO-Ar, the shallowness of the surface gives rise to agreement that is less satisfactory, but a global scaling provides better qualitative agreement. The assignment of the spectra is far from straightforward and is only possible with guidance from the calculated energies and wavefunctions of the energy levels of the complex. Previous assignments are discussed in the light of this conclusion. PMID- 19123504 TI - Binary-collision-induced longitudinal relaxation in gas-phase 83Kr. AB - Density dependent NMR relaxation measurements of noble gases can provide complementary information to that obtained from relaxation studies of molecular gases. However, conventional noble gas NMR is typically hindered by low sensitivity or prohibitively long relaxation times. In this work, the low sensitivity of (83)Kr (I=92) was overcome by spin exchange optical pumping, and the quadrupolar interaction dominated (83)Kr T(1) times of 40-400 s enabled rapid collection of relaxation data. The density dependence of the (83)Kr longitudinal relaxation in pure krypton was found to be about 1.6 x 10(-3) amagat(-1) s(-1). Experiments were also performed in krypton mixtures containing either helium or nitrogen as a buffer gas. By varying the composition and the density of these mixtures, the density dependence of buffer gas induced relaxation and the relaxation efficiency of (83)Kr-buffer gas collisions were determined. The results from these gas mixtures are compared with those from pure krypton. PMID- 19123505 TI - Extended negative glow and "hollow anode" discharges for submillimeter-wave observation of CN-, C2H-, and C4H-. AB - Three molecular anions, CN(-), C(2)H(-), and C(4)H(-), have been detected in an extended negative glow discharge and a "hollow anode" discharge in the submillimeter-wave region. These electrical discharges have been unexpectedly found to be reasonable anion sources. The measurements have been extended up to 830 GHz, and the molecular constants have been improved significantly compared with those determined by microwave and millimeter-wave spectroscopy. The parent gas mixture of C(2)N(2) or C(2)H(2) of 2 mTorr and Ar or Ne buffer of 15 mTorr was used for the production of CN(-) or C(2)H(-) and C(4)H(-). The number densities of the anions in the cell were estimated to be approximately 1x10(9), approximately 3x10(7), and approximately 3x10(6) cm(-3) for CN(-), C(2)H(-), and C(4)H(-), respectively. PMID- 19123506 TI - Pathways and reduced-dimension five-dimensional potential energy surface for the reactions H3+ + CO-->H2+HCO+ and H3+ + CO-->H2+HOC+. AB - To obtain theoretical insight regarding the stability and formation dynamics of the interstellar ions HCO(+) and HOC(+), stationary points and the associated vibrational frequencies on the full nine-dimensional potential energy surface for the electronic ground state have been calculated using coupled-cluster theory with both single and double substitutions (CCSD). The energetics were refined with a higher-level coupled-cluster method CCSD(T), with core-valence electron correlation treated at the complete basis set limit. To elucidate the formation mechanism and internal relaxation processes, the reaction paths for the reactions H(3) (+)+CO-->H(2)+HCO(+) and H(3) (+)+CO-->H(2)+HOC(+) were calculated at the second-order Moller-Plesset (MP2) level, and corresponding single-point energies were obtained at the higher CCSD(T)/aug-cc-pVTZ level. Based on the analysis of the main reaction processes, a reduced-dimension five-dimensional potential energy surface for this system was constructed from 128 440 ab initio points calculated at the CCSD(T)/aug-cc-pVTZ level. PMID- 19123507 TI - Renner-Teller coupled-channel dynamics of the N(2D)+H2 reaction and the role of the NH2 A2A1 electronic state. AB - We present Renner-Teller (RT) and Born-Oppenheimer (BO) coupled-channel (CC) dynamics of the reaction (14)N((2)D)+(1)H(2)(X (1)Sigma(g) (+))-->NH(X (3)Sigma( ))+H((2)S), considering both NH(2) coupled electronic states, X (2)B(1) and A (2)A(1), and Coriolis interactions. We use the best available potential energy surfaces (PESs), and we obtain initial-state-resolved reaction probabilities, cross sections, and rate constants through the real wavepacket and flux methods, taking into account the nuclear-spin statistics for both electronic states. Contrasting RT-CC with more approximate results, we point out the role of RT and Coriolis couplings, and discuss the importance of the A (2)A(1) excited state on the initial-state-resolved dynamics and on the thermal kinetic rate. Confirming the previous results, RT couplings transfer partly the reactivity from X (2)B(1) to A (2)A(1), and CC calculations are necessary to obtain accurate high-energy cross sections. When H(2) is initially rotating, RT couplings enhance strongly the electronic-state-resolved A (2)A(1) reactivity. Considering the nuclear-spin statistics for both electronic states, we find out that the A (2)A(1) state plays a significant role in the rotationally resolved dynamics of N((2)D)+ortho-H(2). However, the BO-X (2)B(1) approximation gives a thermal rate that is slightly smaller than the one obtained by the RT-CC calculations. This implies that this usual approximation is acceptable to calculate unresolved kinetic data of the title reaction. Our calculated rate constant values within the 213-300 K temperature interval are in excellent agreement with the experimental ones. PMID- 19123508 TI - Photoelectron imaging of atomic chlorine and bromine following photolysis of CH2BrCl. AB - Photoionization of chlorine and bromine atoms following photodissociation of CH(2)BrCl was studied in the wavelength range of 231-238 nm by photoelectron imaging technique. Final state-specific speed and angular distributions of the photoelectron were recorded. Analysis of relative branching ratios to different levels of Cl(+) and Br(+) revealed that the final ion level distributions are generally dominated by the preservation of the ion-core configuration of the intermediate resonant state. Some J(c) numbers of the intermediate states were newly assigned according to this regulation. The configuration interaction between resonant states and the autoionization in the continuum were also believed to play an important role in the ionization process since some ions that deviate from the regulation mentioned ahead were observed. The angular distributions of the electrons were found to be well characterized by beta(2) and beta(4), although the ionization process of chlorine and bromine atoms involves three photons. PMID- 19123509 TI - Electron binding motifs in the (CS2)n- (n>4) cluster anions. AB - Photoelectron imaging spectroscopy of (CS(2))(n) (-), n>4, reveals a new state with an electron binding energy smaller than that of any of the corresponding CS(2) (-) and C(2)S(4) (-) states known to date. With support from ab initio calculations, two long-lived, metastable binding motifs with small electron binding energies are discussed for these clusters. The first is a solvent network permeating state, where the excess electron is delocalized over a number of linear CS(2) molecules. The second is an excited (2)B(1) state of the core CS(2) (-) anion stabilized at a slightly bent geometry by the solvation interactions. Based on the observed solvation-induced shifts in binding energy, the second motif is favored. PMID- 19123510 TI - Theoretical study of noble-gas containing metal halides. AB - Equilibrium structures, energetic stability, and vibrational frequencies of noble gas containing metal halides, MNgX and NgMX (Ng=Ar,Kr,Xe; M=Cu,Ag,Au; X=F,Cl,Br) have been studied computationally using coupled cluster, density functional, and perturbation techniques. The NgMX species have been found to be stable with the Ng-M bond dissociation energy of 2-22 kcalmol. Our calculations indicate that the argon-containing MNgX compounds are unstable or very weakly bound. For most of the krypton- and xenon-containing species, well-defined (MNg)(delta+)X(delta-) equilibrium structures have been located. Large MNgX-->Ng+MX reorganization barriers for some of the MNgX molecules (e.g., AuXeF and AuXeCl) indicate their considerable kinetic stability. The presented results suggest that direct observation of the most stable of the MNgX molecules might be possible in experiment. PMID- 19123511 TI - Density functional theory of inhomogeneous liquids. III. Liquid-vapor nucleation. AB - The process of nucleation of vapor bubbles from a superheated liquid and of liquid droplets from a supersaturated vapor is investigated using the modified core van der Waals model density functional theory [J. F. Lutsko, J. Chem. Phys. 128, 184711 (2008)]. A novel approach is developed whereby nucleation is viewed as a transition from a metastable state to a stable state via the minimum free energy path, which is identified using the nudged elastic-band method for exploring free energy surfaces. This allows for the unbiased calculation of the properties of sub- and super-critical clusters, as well as of the critical cluster. For Lennard-Jones fluids, the results compare well to simulation and support the view that at low to moderate supersaturation nucleation proceeds smoothly rather than via spinodal-like instabilities as has been suggested recently. The possibility that the nucleation barrier vanishes at high supersaturation is, however, not ruled out. PMID- 19123512 TI - Thermodynamic properties of double square-well fluids: computer simulations and theory. AB - Computer simulations have been performed to obtain the thermodynamic properties of fluids with double square-well potentials, that is, potentials consisting of two adjacent square wells with different depths. The compressibility factor, excess energy, chemical potential, constant-volume excess heat capacity, and other derived properties have been obtained. These data have been used to test the performance of several perturbation theories for predicting the thermodynamic properties of this kind of fluids. Good agreement is found on the whole between theory and simulation at supercritical temperatures. The possible presence of anomalous behavior at high densities in the fluids considered has been also analyzed in light of the same theories, with the result that in general, they do not predict such anomalous behavior, with the possible exception of a Monte Carlo based perturbation theory for relatively large potential widths at high densities and very low temperatures. PMID- 19123513 TI - Molecular dynamics simulations of AOT-water/formamide reverse micelles: structural and dynamical properties. AB - We present results from molecular dynamics simulations performed on reverse micelles immersed in cyclohexane. Three different inner polar phases are considered: water (W), formamide (FM), and an equimolar mixture of the two solvents. In all cases, the surfactant was sodium bis(2-ethylhexyl) sulfosuccinate (usually known as AOT). The initial radii of the micelles were R approximately 15 A, while the corresponding polar solvent-to-surfactant molar ratios were intermediate between w(0)=4.3 for FM and w(0)=7 for W. The resulting overall shapes of the micelles resemble distorted ellipsoids, with average eccentricities of the order of approximately 0.75. Moreover, the pattern of the surfactant layer separating the inner pool from the non-polar phase looks highly irregular, with a roughness characterized by length scales comparable to the micelle radii. Solvent dipole orientation polarization along radial directions exhibit steady growths as one moves from central positions toward head group locations. Local density correlations within the micelles indicate preferential solvation of sodium ionic species by water, in contrast to the behavior found in bulk equimolar mixtures. Still, a sizable fraction of approximately 90% of Na(+) remains associated with the head groups. Compared to bulk results, the translational and rotational modes of the confined solvents exhibit important retardations, most notably those operated in rotational motions where the characteristic time scales may be up to 50 times larger. Modifications of the intramolecular connectivity expressed in terms of the average number of hydrogen bonds and their lifetimes are also discussed. PMID- 19123514 TI - Theory and atomistic simulation of krypton fluid. AB - An ab initio interaction potential available in literature is scaled via an empirical procedure and used in an extensive computer simulation study to investigate the thermodynamic properties and self-diffusion coefficient of krypton over a wide range of densities and temperatures. The thermodynamic properties of the fluid phase equilibriums are computed utilizing the Gibbs ensemble Monte Carlo simulation technique. The equation of state and the pair correlation function are obtained using the NVT-Monte Carlo simulation method. The time-correlation function formalism of Green-Kubo is applied in molecular dynamics simulations to calculate the self-diffusion coefficient. Furthermore, the modified Cohen-Turnbull theory is employed to determine the self-diffusion coefficient and the mean free volume needed for this purpose is provided via the generic van der Waals theory. The virial minimization method is used to compute the effective diameter and the results are applied within the generic van der Waals theory as the repulsion-attraction splitting distance of the interaction potential. A remarkable agreement is observed between the computed and empirical results for the orthobaric densities, the vapor pressure, the critical point, and the equation of state. A detailed analysis is presented for the calculated self diffusion coefficient. PMID- 19123515 TI - Dipole allowed transitions in GdF: A four-component relativistic general open shell configuration interaction study. AB - A four-component relativistic study of electronic transitions in the gadolinium monofluoride molecule (GdF) is presented. The electronic spectra of GdF have been investigated with a general open-shell configuration interaction method, where active electrons are distributed among molecular spinors mainly consisting of the Gd 4f, 5d, and 6s atomic spinors. The near-degeneracy effects of these spinors on the molecular electronic structure are considered by the valence full-CI-like approach. By the magnitudes of calculated transition dipole moments, the candidates for the observable transitions were selected. The present result is complementary to our previous study based on multireference configuration interaction singles and doubles calculations, which identified the electronic excited states of GdF by comparing the calculated excitation energies and angular momenta with those given by the laser spectroscopy. The spectra of the excited states less than 3.0 eV have been refined with the help of the calculated transition probabilities. The transitions between the excited states are newly analyzed and a rearrangement is proposed. PMID- 19123516 TI - Predictions of melting, crystallization, and local atomic arrangements of aluminum clusters using a reactive force field. AB - A parametrized reactive force field model for aluminum ReaxFF(Al) has been developed based on density functional theory (DFT) data. A comparison has been made between DFT and ReaxFF(Al) outputs to ascertain whether ReaxFF(Al) is properly parametrized and to check if the output of the latter has correlation with DFT results. Further checks include comparing the equations of state of condensed phases of Al as calculated from DFT and ReaxFF(Al). There is a good match between the two results, again showing that ReaxFF(Al) is correctly parametrized as per the DFT input. Simulated annealing has been performed on aluminum clusters Al(n) using ReaxFF(Al) to find the stable isomers of the clusters. A plot of stability function versus cluster size shows the existence of highly stable clusters (magic clusters). Quantum mechanically these magic clusters arise due to the complete filling of the orbital shells. However, since force fields do not care about electrons but work on the assumption of validity of Born-Oppenheimer approximation, the magic clusters are therefore correlated with high structural symmetry. There is a rapid decline in surface energy contribution due to the triangulated nature of the surface atoms leading to higher coordination number. The bulk binding energy is computed to be 76.8 kcal/mol. This gives confidence in the suitability of ReaxFF for studying and understanding the underlying dynamics in aluminum clusters. In the quantification of the growth of cluster it is seen that as the size of the clusters increase there is preference for the coexistence of fcc/hcp orders at the expense of simple icosahedral ordering, although there is some contribution from distorted icosahedral ordering. It is found that even for aluminum clusters with 512 atoms distorted icosahedral ordering exists. For clusters with N>/=256 atoms fcc ordering dominates, which implies that at this point we are already on the threshold of bulklike bonding. PMID- 19123517 TI - Electron density topology of cubic structure I Xe clathrate hydrate at high pressure. AB - In this report, we present a detailed powder x-ray diffraction study of the structural properties and charge density topology of structure I Xe clathrate hydrate under high pressure and room temperature. The pressure dependence of the structural parameters was determined by applying a Rietveld analysis to the experimental data. The combined Rietveld/maximum entropy method was used to derive the most probable charge density distribution at each pressure. Our results show that the charge density distribution of the encaged Xe atoms differs depending on the type of host cage at all pressures. Spherical electron density distributions were observed for the Xe atoms in the small cages while the atoms in the large cages showed longitudinal elongated electronic distributions. Along with the observed cage deformations, the change in electronic density distribution represents a clear indication that the guest-host interaction differs significantly between the small and large cages at high pressures. A similar behavior has been previously reported in low-temperature studies of methane clathrate hydrate. PMID- 19123518 TI - Thermodynamic and transport properties of the Weeks-Chandler-Andersen fluid: theory and computer simulation. AB - The equilibrium molecular dynamics (MD) simulation method is utilized to compute the equation of state, pair correlation function, and transport coefficients of the Weeks-Chandler-Andersen model fluid over a wide range of densities and temperatures. The calculated pair correlation function is used within the virial minimization method to obtain the effective diameter and the results are applied within the generic van der Waals (GvdW) theory to compute the GvdW parameters. The GvdW parameter B, which corresponds to the mean excluded volume of particles, is used to calculate the mean free volume. The computed effective diameter and Gvdw parameters are then fitted to functions of the density and temperature. The modified Cohen-Turnbull theory is employed to calculate the diffusion coefficient using the computed effective diameter and mean free volume. A detailed analysis is presented for the results of the theory and MD simulations for the diffusion coefficient. The system-size dependence of the diffusion coefficient through the MD simulation is studied. The Dullien formula is used to obtain the shear viscosity and the results are compared with those from MD simulations. PMID- 19123519 TI - The Ornstein-Zernike equation and critical phenomena in fluids. AB - It is shown that there are two classes of closure equations for the Ornstein Zernike (OZ) equation: the analytical equations B=B((an)) type of hyper-netted chain approximation, Percus-Yevick approximation etc., and the nonanalytical equation B=B((non)), where B((nan))=B((RG))+B((cr)); B((RG)) is the regular (analytical) component of the bridge functional, and B((cr)) is the critical (nonanalytical) component of B((nan)). The closure equation B((an)) defines coordinates of a critical point and other individual features of critical phenomena, and B((nan)) defines known relations between critical exponents. It is shown that a necessary condition for the existence of a nonanalytical solution of the OZ equation is the equality 5-eta=delta(1+eta), where eta and delta are critical exponents, the values of which can change in a narrow interval. It is shown that the transition from analytical solution to nonanalytical solution is accompanied by a step of derivative of pressure. On the phase diagram of fluids the boundaries dividing the area of existence of analytical and nonanalytical solutions are indicated. PMID- 19123520 TI - Translation-rotation decoupling of colloidal clusters of various symmetries. AB - Single-particle tracking was used to measure the diffusion in aqueous suspension of dilute colloidal clusters, 2-9 mum in size, fabricated by joining 1.57 mum spheres into planar arrangements of various particle numbers and symmetries (doublet, trimer, square, pentamer, hexamer, and so forth). They were allowed to sediment close to a glass surface and their Brownian motion parallel to the surface, effectively in a two dimensional (2D) geometry, was imaged in a microscope in the presence of 3 mM monovalent salt to essentially screen electrostatic interactions. Geometric asymmetry produced systematically increasing discrepancy between the equivalent hydrodynamic radius of translation and rotation-tabulated in this paper. Our observations include cases where the effective hydrodynamic radius changes more rapidly for translation than rotation, the converse, and also cases where the effective hydrodynamic radius for translation changes significantly, while that of rotation is effectively constant. The significance is to document the connection between translational and rotational 2D mobilities for geometrical shapes not described by the Stokes Einstein-Debye equations for spherical particles. PMID- 19123521 TI - Probing dynamics of single molecules: Nonlinear spectroscopy approach. AB - A two level model of a single molecule undergoing spectral diffusion dynamics and interacting with a sequence of two short laser pulses is investigated. Analytical solution for the probability of n=0,1,2 photon emission events for the telegraph and Gaussian processes is obtained. We examine under what circumstances the photon statistics emerging from such pump-probe setup provides new information on the stochastic process parameters and what are the measurement limitations of this technique. The impulsive and selective limits, the semiclassical approximation, and the fast modulation limit exhibit general behaviors of this new type of spectroscopy. We show that in the fast modulation limit, where one has to use impulsive pulses in order to obtain meaningful results, the information on the photon statistics is contained in the molecule's dipole correlation function, equivalently to continuous wave experiments. In contrast, the photon statistics obtained within the selective limit depends on the both spectral shifts and rates and exhibits oscillations, which are not found in the corresponding line shape. PMID- 19123522 TI - Freezing of charged colloids in slit pores. AB - Using Monte Carlo simulations in the grand canonical and isobaric ensembles we investigate freezing phenomena in a charged colloidal suspension confined to narrow slit pores. Our model involves only the macroions which interact via a Derjaguin-Landau-Verwey-Overbeek (DLVO) potential supplemented by a soft-sphere potential. We focus on DLVO parameters typical for moderately charged silica particles (with charges Z approximately 35) in solvents of low ionic strengths. The corresponding DLVO interactions are too weak to drive a (bulk) freezing transition. Nevertheless, for sufficiently small surface separations L(z) the confined systems display not only layering but also significant in-plane crystalline order at chemical potentials where the bulk system is a globally stable fluid (capillary freezing). At confinement conditions related to two-layer systems the observed in-plane structures are consistent with those detected in ground state calculations for perfect Yukawa bilayers [R. Messina and H. Lowen, Phys. Rev. Lett. 91, 146101 (2003)]. Here we additionally observe (at fixed L(z)) a compression-induced first-order phase transition from a two-layer to a three layer system with different in-plane structure, in agreement with previous findings for pure hard spheres. PMID- 19123524 TI - Dynamical simulations of charged soliton transport in conjugated polymers with the inclusion of electron-electron interactions. AB - We present numerical studies of the transport dynamics of a charged soliton in conjugated polymers under the influence of an external time-dependent electric field. All relevant electron-phonon and electron-electron interactions are nearly fully taken into account by simulating the monomer displacements with classical molecular dynamics and evolving the wave function for the pi electrons by virtue of the adaptive time-dependent density matrix renormalization group simultaneously and nonadiabatically. It is found that after a smooth turn on of the external electric field the charged soliton is accelerated at first up to a stationary constant velocity as one entity consisting of both the charge and the lattice deformation. An Ohmic region (6 mV/A20 kHz) and magnetic fields (750-900 MHz (1)H frequencies) using moderate rf fields, exhibit numerous cross peaks corresponding to long (up to 6-7 A) (13)C-(13)C distances which are particularly useful in protein structure determination. Using results from PAR spectra we calculate the structure of the Crh protein. PMID- 19123540 TI - Cavity-dumped 2.70 microm erbium laser using optomechanical shutter. AB - A cavity-dumped 2.70 microm erbium laser with a frustrated total internal reflection (FTIR) shutter was investigated and compared with a Q-switched erbium laser using the FTIR shutter. The Q-switched and the cavity-dumped 2.70 microm laser outputs were obtained with a dichroic coated mirror with high reflectance at 2.70 microm and high transmittance at 2.79 microm. For the Q-switched operation, a maximum peak power of 33.5 kW was achieved, and its pulse width was 1.3 mus. For the cavity-dumped operation, the laser pulse energy was optimized by changing the switching time of the FTIR shutter. When the pulse width is reduced to 210 ns, the peak power increases to 154 kW. PMID- 19123541 TI - New versatile and linear optical sensor based on electro-optical modulation and compensation. AB - This paper is devoted to the description and characterization of a new electro optic sensor based on a polarimetric device operating in reflection mode, and on a closed-loop controller. The main features of the proposed sensor are versatility, insensitivity to the laser beam optical power fluctuation, and fully linear working mode. The resolution of the sensor, in terms of phase shift, is shown to be better than +/-20 mrad and its accuracy to +/-50 mrad. PMID- 19123542 TI - High-density, high-temperature alkali vapor cell. AB - We present a vapor cell design that enables resonant optical spectroscopy in high density, high-temperature alkali metal vapors. Optical access is provided via reflection from the interface between the vapor and a sapphire window. The cell resists corrosion from the highly reactive alkali vapors, in our case, potassium, up to 800 degrees C (number densities up to 10(19) cm(-3)). The cells maintain their integrity for an average of 100 h above 500 degrees C with about 10 heating/cooling cycles. PMID- 19123543 TI - Imaging piezospectroscopy. AB - A novel instrument is described for obtaining accurate high-resolution residual stress images of aluminum oxide materials, based on the piezospectroscopy of Cr(3+) dopant ions. The instrument employs a charge coupled device camera, a narrow bandpass tunable filter and the use of Tikhonov regularization for reconstruction of the raw spectral data. The experimental accuracy and spectral shift resolution of this method were analyzed with two calibration light sources and with independently measured spectra, and were found to be approximately +/ 0.01 nm across the pixel array. This is close to the theoretically obtainable accuracy for the particular filter used, a solid etalon Fabry-Perot filter with a passband of 0.25 nm, based on an analysis with simulated data. The spectral resolution corresponds to a stress resolution, under biaxial stress conditions, of +/-40 MPa. PMID- 19123544 TI - High-resolution Rydberg tagging time-of-flight measurements of atomic photofragments by single-photon vacuum ultraviolet laser excitation. AB - By coupling a comprehensive tunable vacuum ultraviolet (VUV) laser system to a velocity-mapped ion imaging apparatus, we show that high-resolution high-n Rydberg tagging time-of-flight (TOF) measurements of nascent atomic photofragments formed by laser photodissociation can be made using single-photon VUV laser photoexcitation. To illustrate this single-photon Rydberg tagging TOF method, we present here the results of the VUV laser high-n Rydberg tagging TOF measurements of O((3)P(2)) and S((3)P(2)) formed in the photodissociation of SO(2) and CS(2) at 193.3 and 202.3 nm, respectively. These results are compared to those obtained by employing the VUV laser photoionization time-sliced velocity mapped ion imaging technique. The fact that the kinetic energy resolutions achieved in the VUV laser high-n Rydberg tagging TOF measurements of O and S atoms are found to be higher than those observed in the VUV laser photoionization, time-sliced velocity-mapped ion imaging studies show that the single-photon VUV laser high-n Rydberg tagging TOF method is useful and complementary to state-of-the-art time-sliced velocity-mapped ion imaging measurements of heavier atomic photofragments, such as O and S atoms. Furthermore, the general agreement observed between the VUV laser high-n Rydberg tagging TOF and velocity-mapped ion imaging experiments supports the conclusion that the lifetimes of the tagged Rydberg states of O and S atoms are sufficiently long to allow the reliable determination of state-resolved UV photodissociation cross sections of SO(2) and CS(2) by using the VUV laser high-n Rydberg tagging TOF method. PMID- 19123545 TI - A time-resolved single-pass technique for measuring optical absorption coefficients of window materials under 100 GPa shock pressures. AB - An experimental method was developed to perform time-resolved, single-pass optical absorption measurements and to determine absorption coefficients of window materials under strong shock compression up to approximately 200 GPa. Experimental details are described of (i) a configuration to generate an in situ dynamic, bright, optical source and (ii) a sample assembly with a lithium fluoride plate to essentially eliminate heat transfer from the hot radiator into the specimen and to maintain a constant optical source within the duration of the experiment. Examples of measurements of optical absorption coefficients of several initially transparent single crystal materials at high shock pressures are presented. PMID- 19123546 TI - Near UV-near IR Fourier transform spectrometer using the beam-folding position tracking method based on retroreflectors. AB - A near UV-near IR Fourier transform spectrometer based on a beam-folding position tracking method realized by using retroreflectors is reported. The use of retroreflectors maintains all beams in the beam-fold arrangement in parallel with the incident beams. The beam-folding interferometer used for position tracking is arranged to have optical path symmetry with the measurement interferometer in the zero path difference position of the measurement interferometer, and the vertex of the movable retroreflector in the measurement interferometer is arranged very close to the midpoint of the vertices of two movable retroreflectors in the position-tracking interferometer. These measures keep the equivalent optical axis of the position-tracking interferometer well in line with that of the measurement interferometer even with translational misalignments. Therefore, the change in the optical path difference of the position-tracking interferometer is always synchronous to that of the measurement interferometer during the scanning process. That is, the position-tracking error can be suppressed to very small values during a scan. We have demonstrated a UV-near IR Fourier transform spectrometer with a standard quality ball-bearing translation stage achieving a resolution close to the theoretical resolution of approximately 0.28 cm(-1) at the He-Ne laser wavelength when the scan distance reaches the travel distance of over 2 cm. This was achieved without the need for elaborate optics, sophisticated detecting electronics, and high-precision servomotion control. PMID- 19123547 TI - A new titration system of a novel split-type superconducting magnet NMR spectrometer. AB - A new titration system for studying protein-ligand interactions has been developed. In this system, the sample solution is circulated in the route formed by an access path in a split superconducting magnet to maintain a constant protein concentration during the titration experiments. A concentration-control procedure for the ligand/protein ratio is devised, and the ligand/protein ratio is well controlled by this apparatus. PMID- 19123548 TI - A broadband absorption spectrometer using light emitting diodes for ultrasensitive, in situ trace gas detection. AB - A broadband absorption spectrometer has been developed for highly sensitive and target-selective in situ trace gas measurements. The instrument employs two distinct modes of operation: (i) broadband cavity enhanced absorption spectroscopy (BBCEAS) is used to quantify the concentration of gases in sample mixtures from their characteristic absorption features, and (ii) periodic measurements of the cavity mirrors' reflectivity are made using step-scan phase shift cavity ringdown spectroscopy (PSCRDS). The latter PSCRDS method provides a stand-alone alternative to the more usual method of determining mirror reflectivities by measuring BBCEAS absorption spectra for calibration samples of known composition. Moreover, the instrument's two modes of operation use light from the same light emitting diode transmitted through the cavity in the same optical alignment, hence minimizing the potential for systematic errors between mirror reflectivity determinations and concentration measurements. The ability of the instrument to quantify absorber concentrations is tested in instrument intercomparison exercises for NO(2) (versus a laser broadband cavity ringdown spectrometer) and for H(2)O (versus a commercial hygrometer). A method is also proposed for calculating effective absorption cross sections for fitting the differential structure in BBCEAS spectra due to strong, narrow absorption lines that are under-resolved and hence exhibit non-Beer-Lambert law behavior at the resolution of the BBCEAS measurements. This approach is tested on BBCEAS spectra of water vapor's 4v+delta absorption bands around 650 nm. The most immediate analytical application of the present instrument is in quantifying the concentration of reactive trace gases in the ambient atmosphere. The instrument's detection limits for NO(3) as a function of integration time are considered in detail using an Allan variance analysis. Experiments under laboratory conditions produce a 1sigma detection limit of 0.25 pptv for a 10 s acquisition time, which improves with further signal averaging to 0.09 pptv in 400 s. Finally, an example of the instrument's performance under field work conditions is presented, in this case of measurements of the sum of NO(3)+N(2)O(5) concentrations in the marine boundary layer acquired during the Reactive Halogens in the Marine Boundary Layer field campaign. PMID- 19123549 TI - A newly designed single etalon double edge Doppler wind lidar receiving optical system. AB - A new optical Doppler lidar receiving system which can measure Doppler-shifted incoherent scattering light has been proposed and tested. This system functions in a manner similar to two edge filters and consists of a single etalon, a polarization beam splitter, and mirrors that can guide scattered light into a single etalon with two different angles. These two incident angles are precisely adjusted by the polarization beam splitter and mirrors. Using this optical receiving system we can measure a moving target with a high sensitivity. Theoretical calculations show that this system will have a better optical sensitivity than a single edge system and will allow the use of incoherent Doppler lidar for wind velocity measurements. PMID- 19123550 TI - A short working distance multiple crystal x-ray spectrometer. AB - For x-ray spot sizes of a few tens of microns or smaller, a millimeter-sized flat analyzer crystal placed approximately 1 cm from the sample will exhibit high energy resolution while subtending a collection solid angle comparable to that of a typical spherically bent crystal analyzer (SBCA) at much larger working distances. Based on this observation and a nonfocusing geometry for the analyzer optic, we have constructed and tested a short working distance (SWD) multicrystal x-ray spectrometer. This prototype instrument has a maximum effective collection solid angle of 0.14 sr, comparable to that of 17 SBCA at 1 m working distance. We find good agreement with prior work for measurements of the Mn Kbeta x-ray emission and resonant inelastic x-ray scattering for MnO, and also for measurements of the x-ray absorption near-edge structure for Dy metal using Lalpha(2) partial-fluorescence yield detection. We discuss future applications at third- and fourth-generation light sources. For concentrated samples, the extremely large collection angle of SWD spectrometers will permit collection of high-resolution x-ray emission spectra with a single pulse of the Linac Coherent Light Source. The range of applications of SWD spectrometers and traditional multi-SBCA instruments has some overlap, but also is significantly complementary. PMID- 19123551 TI - Scalable time-correlated photon counting system with multiple independent input channels. AB - Time-correlated single photon counting continues to gain importance in a wide range of applications. Most prominently, it is used for time-resolved fluorescence measurements with sensitivity down to the single molecule level. While the primary goal of the method used to be the determination of fluorescence lifetimes upon optical excitation by short light pulses, recent modifications and refinements of instrumentation and methodology allow for the recovery of much more information from the detected photons, and enable entirely new applications. This is achieved most successfully by continuously recording individually detected photons with their arrival time and detection channel information (time tagging), thus avoiding premature data reduction and concomitant loss of information. An important property of the instrumentation used is the number of detection channels and the way they interrelate. Here we present a new instrument architecture that allows scalability in terms of the number of input channels while all channels are synchronized to picoseconds of relative timing and yet operate independent of each other. This is achieved by means of a modular design with independent crystal-locked time digitizers and a central processing unit for sorting and processing of the timing data. The modules communicate through high speed serial links supporting the full throughput rate of the time digitizers. Event processing is implemented in programmable logic, permitting classical histogramming, as well as time tagging of individual photons and their temporally ordered streaming to the host computer. Based on the time-ordered event data, any algorithms and methods for the analysis of fluorescence dynamics can be implemented not only in postprocessing but also in real time. Results from recently emerging single molecule applications are presented to demonstrate the capabilities of the instrument. PMID- 19123552 TI - Simple two-dimensional-imaging spectrograph with wedged narrow band filters. AB - A minute, yet practical, modification of a well-known spectrograph based on a set of wedged narrow band filters is presented. It makes possible two-dimensional imaging spectral measurements with a potential subpixel precision of a few micrometers. The simple spectroscopic device can evaluate dispersion of quantities which can be transformed into an image, e.g., into a spatial beam distribution. The spectrograph was used to measure angular dispersion of both a laser oscillator producing femtosecond pulses and a simple dispersive optical system, an optical wedge. PMID- 19123553 TI - Velocity map imaging in time of flight mass spectrometry. AB - A new variation on time of flight mass spectrometry is presented, which uses a fast framing charge coupled device camera to velocity map image multiple product masses in a single acquisition. The technique is demonstrated on two photofragmentation processes, those of CS(2) and CH(3)S(2)CH(3) (dimethyldisulfide) at a photolysis wavelength of 193 nm. In both cases, several mass fragments are imaged simultaneously, and speed distributions and anisotropy parameters are extracted that are comparable to those obtained by imaging each fragment separately in conventional velocity map imaging studies. PMID- 19123554 TI - Transferring cold atoms in double magneto-optical trap by a continuous-wave transfer laser beam with large red detuning. AB - A novel scheme of transferring cold atoms in a double magneto-optical trap (MOT) system has been experimentally demonstrated. Cold cesium atoms trapped in a vapor cell MOT are efficiently transferred to an ultrahigh-vacuum (UHV) MOT by a continuous-wave divergent Gaussian transfer laser beam. When large red detuning and moderate intensity are adopted for the transfer laser beam, enhancement of the recapturing of atoms in the UHV MOT is clearly observed. Using the divergent transfer laser beam (diameter of approximately 1.60 mm in the vapor-cell MOT region) with typical power of approximately 20.2 mW, up to approximately 85% of transfer efficiency is obtained when the frequency detuning is set to around -1.2 GHz, and it is not sensitive to small detuning variation. This transfer is much efficient compared with that in the case of continuous-wave near-resonance weak transfer laser beam (typical power of order of approximately 100 microW and typical frequency detuning of approximately-10 MHz) which is normally used in double-MOT experiment. The enhancement is ascribed to the guiding effect on cold atomic flux by transverse dipole potential of the large red-detuned transfer laser beam. PMID- 19123555 TI - A new spin-polarized photoemission spectrometer with very high efficiency and energy resolution. AB - A new spin- and angle-resolved photoemission spectrometer was developed adopting the very-low-energy-electron-diffraction (VLEED)-type spin polarimeter. The Fe(001)p(1x1)-O film grown on MgO(001) crystal for the VLEED target yields significantly high spin-resolving power, the effective Sherman function of 0.40+/ 0.02, with long lifetime and stability compared to the conventional Fe(001) target. Under the favor of high resolving power, approximately 100 times higher efficiency than that of conventional Mott-type spin polarimeter, the figure of merit of 1.9+/-0.2x10(-2) was achieved. Owing to this high efficiency, high energy resolution can be realized with this new spin-polarized photoemission spectrometer. The simplified ways of target preparation and revitalization make the VLEED spin polarimeter much more convenient and feasible for the spin polarized photoemission spectroscopy. PMID- 19123556 TI - Simulation and analysis of solenoidal ion sources. AB - We present a detailed analysis and simulation of solenoidal, magnetically confined electron bombardment ion sources, aimed at molecular beam detection. The aim is to achieve high efficiency for singly ionized species while minimizing multiple ionization. Electron space charge plays a major role and we apply combined ray tracing and finite element simulations to determine the properties of a realistic geometry. The factors controlling electron injection and ion extraction are discussed. The results from simulations are benchmarked against experimental measurements on a prototype source. PMID- 19123557 TI - Design and characterization of combined function multipole magnet for accelerators. AB - This paper presents the design and analysis of a multipurpose combined function magnet for use in accelerators. This magnet consists of three corrector magnets: (i) skew quadrupole, (ii) horizontal dipole, and (iii) vertical dipole magnets, along with the main sextupole magnet. The strength of the corrector magnets is smaller than that of the main sextupole magnet. The strength of all the four magnets can be varied independently. The excitation strength required to produce skew quadrupole gradient and the presence of various multipole components in the magnet are estimated using first order perturbation theory. The experimental data for the variation of the sextupole strength and its higher order multipoles in the presence of skew quadrupole excitations are presented and compared to the theoretical predictions. Simulation using two-dimensional fine element code, Poisson, is also done. Results obtained from all the above three methods are found to be in good agreement with each other. The variations of skew quadrupole gradient for different sextupole excitations are also measured. The validity of this theory is also checked for various combinations of excitations including the case where magnet gets saturated. The excitation strengths required for producing the horizontal and vertical dipole fields are estimated analytically along with the presence of various multipoles. Theoretical predictions of permissible multipoles are compared to the results obtained from simulation. PMID- 19123558 TI - Correction of spherical aberration for an electrostatic gridded lens. AB - Two methods to correct spherical aberration in electrostatic gridded lenses have been studied using ray-tracing simulations. Both methods are based on modifying the electrostatic field on the radial periphery of the lens. In the simplest case, the modification is done by extending the grid support axially. In the second method, the electric field on a radial periphery of the lens is modified by applying optimum voltage on an isolated correcting electrode. It is demonstrated that, for a given focal length, the voltage on this lens can be optimized for minimum aberration, and also that these lenses reduce the emittance growth of the ion beam. PMID- 19123559 TI - High precision high voltage divider and its application to electron beam ion traps. AB - A high precision high voltage divider has been developed for the electron beam ion trap in Shanghai. The uncertainty caused by the temperature coefficient of resistance (TCR) and the voltage coefficient of resistance has been studied in detail and was minimized to the level of ppm (10(-6)) range. Once the TCR was matched between the resistors, the precision of the dividing ratio finally reached the ppm range also. We measured the delay of the divider caused by the capacitor introduced to minimize voltage ripple to be 2.35 ms. Finally we applied the divider to an experiment to measure resonant energies for some dielectronic recombination processes for highly charged xenon ions. The final energies include corrections for both space charge and fringe field effects are mostly under 0.03%. PMID- 19123560 TI - Development of compact size penning ion source for compact neutron generator. AB - For long-life operation, easy to mount and compact in size penning type ion sources are widely used in different fields of research such as neutron generators, material research, and surface etching. One penning type ion source has been developed in our laboratory. Applying high voltage of 2 kV between two oppositely biased electrodes and using permanent magnet of 500 gauss magnetic field along the axis, we had produced the glow discharge in the plasma region. The performance of this source was investigated using nitrogen gas. Deuterium ions were produced and extracted on the basis of chosen electrodes and the angle of extraction. Using a single aperture plasma electrode, the beam was extracted along the axial direction. The geometry of plasma electrode is an important factor for the efficient extraction of the ions from the plasma ion source. The extracted ion current depends upon the shape of the plasma meniscus. A concave shaped plasma meniscus produces converged ion beam. The convergence of extracted ions is related to the extraction electrode angle. The greater the angle, the more the beam converges. We had studied experimentally this effect with a compact size penning ion source. The detailed comparison among the different extraction geometry and different electrode angle are discussed in this paper. PMID- 19123561 TI - A collective scattering system for measuring electron gyroscale fluctuations on the National Spherical Torus Experiment. AB - A collective scattering system has been installed on the National Spherical Torus Experiment (NSTX) to measure electron gyroscale fluctuations in NSTX plasmas. The system measures fluctuations with k( perpendicular)rho(e) less, similar0.6 and k( perpendicular) less, similar20 cm(-1). Up to five distinct wavenumbers are measured simultaneously, and the large toroidal curvature of NSTX plasmas provides enhanced spatial localization. Steerable optics can position the scattering volume throughout the plasma from the magnetic axis to the outboard edge. Initial measurements indicate rich turbulent dynamics on the electron gyroscale. The system will be a valuable tool for investigating the connection between electron temperature gradient turbulence and electron thermal transport in NSTX plasmas. PMID- 19123562 TI - Phase-type quantum-dot-array diffraction grating. AB - A novel phase-type quantum-dot-array diffraction grating (QDADG) is reported. In contrast to an earlier amplitude-type QDADG [C. Wang et al., Rev. Sci. Instrum. 78, 053503 (2007)], the new phase-type QDADG would remove the zeroth order diffraction at some certain wavelength, as well as suppressing the higher-order diffractions. In this paper, the basic concept, the fabrication, the calibration techniques, and the calibration results are presented. Such a grating can be applied in the research fields of beam splitting, laser probe diagnostics, and so on. PMID- 19123563 TI - Improvement of temporal resolution for three-dimensional continuous-wave electron paramagnetic resonance imaging. AB - This paper describes improved temporal resolution for three-dimensional (3D) continuous-wave electron paramagnetic resonance (EPR) imaging. To improve temporal resolution, the duration of magnetic filed scanning that is used to obtain an EPR spectrum for each projection was reduced to 40 ms. The Helmholtz coil pair for field scanning was driven by triangular waves. The uniform distribution of projections was also used to reduce the number of projections for 3D image reconstruction. The reduction reaction of 4-hydroxy-2,2,6,6-tetramethyl piperidinooxy with ascorbic acid was visualized by improved 3D EPR imaging techniques with a temporal resolution of 5.8 s. PMID- 19123564 TI - Development of new in situ observation system for dynamic study of lubricant molecules on metal friction surfaces by two-dimensional fast-imaging Fourier transform infrared-attenuated total reflection spectrometer. AB - To observe the time-dependent two-dimensional (2D) images of spatial distribution of chemically/physically modified lubricant molecules on the metal surface during friction motion, a new in situ technique has been developed by combining the 2D fast-imaging Fourier-transform infrared-attenuated total reflection spectrometer with the temperature-controlled friction equipment containing lubricant agent. Using this new instrument, the time-dependent changes in lubricant molecules, for example, cis-trans isomerization, stress-induced molecular deformation, etc., can be detected successfully. The characteristic features of this instrument have been demonstrated in a detailed and concrete manner by demonstrating the experimental data measured for oleic acid and tricresyl phosphate. PMID- 19123565 TI - Photothermal excitation and laser Doppler velocimetry of higher cantilever vibration modes for dynamic atomic force microscopy in liquid. AB - The authors present an optically based method combining photothermal excitation and laser Doppler velocimetry of higher cantilever vibration modes for dynamic atomic force microscopy in liquid. The frequency spectrum of a silicon cantilever measured in water over frequencies ranging up to 10 MHz shows that the method allows us to excite and detect higher modes, from fundamental to fifth flexural, without enhancing spurious resonances. By reducing the tip oscillation amplitude using higher modes, the average tip-sample force gradient due to chemical bonds is effectively increased to achieve high-spatial-resolution imaging in liquid. The method's performance is demonstrated by atomic resolution imaging of a mica surface in water obtained using the second flexural mode with a small tip amplitude of 99 pm; individual atoms on the surface with small height differences of up to 60 pm are clearly resolved. PMID- 19123566 TI - Single-sided radio-frequency field gradient with two unsymmetrical loops: Applications to nuclear magnetic resonance. AB - Magnetic field gradients are nowadays indispensable to most nuclear magnetic resonance experiments and are at the basis of magnetic resonance imaging (MRI). Most of the time, gradients of the static magnetic field are employed. Gradients of the radio-frequency (rf) field may constitute an interesting alternative. Until now, they were produced by a single loop. We demonstrate in this paper how two unsymmetrical series loops can be optimized to produce rf gradients of much better performances. This optimization is based on a thorough theoretical approach and the gradient uniformity is studied through accurate simulations. Two prototypes were devised: one for a 2.34 T horizontal magnet (used in MRI), and the other for a 4.7 T vertical magnet (used for pure spectroscopic applications). These two-loop systems were designed for proton resonance frequencies (100 and 200 MHz, respectively). Performances of both systems were verified (versus theoretical predictions) by means of experiments employing gradients in view of the determination of the self-diffusion coefficients of liquids. PMID- 19123567 TI - Digital control of force microscope cantilevers using a field programmable gate array. AB - This report describes a cantilever controller for magnetic resonance force microscopy based on a field programmable gate array, along with the hardware and software used to integrate the controller into an experiment. The controller is assembled from a low-cost commercially available software defined radio device and libraries of open-source software. The controller includes a digital filter comprising two cascaded second-order sections ("biquads"), which together can implement transfer functions for optimal cantilever controllers. An appendix in this report shows how to calculate filter coefficients for an optimal controller from measured cantilever characteristics. The controller also includes an input multiplexer and adder used in calibration protocols. Filter coefficients and multiplexer settings can be set and adjusted by control software while an experiment is running. The input is sampled at 64 MHz; the sampling frequency in the filters can be divided down under software control to achieve a good match with filter characteristics. Data reported here were sampled at 500 kHz, chosen for acoustic cantilevers with resonant frequencies near 8 kHz. Inputs are digitized with 12 bit resolution, and outputs are digitized with 14 bits. The experiment software is organized as a client and server to make it easy to adapt the controller to different experiments. The server encapsulates the details of controller hardware organization, connection technology, filter architecture, and number representation. The same server could be used in any experiment, while a different client encodes the particulars of each experiment. PMID- 19123568 TI - Nanoscale lithography with frequency-modulation atomic force microscopy. AB - A lithographic method to draw nanoscale structures by repetitive depositions of the tip material by field evaporation has been developed based on frequency modulation atomic force microscopy (FM-AFM). Because of high stiffness of quartz tuning forks, a force sensor in the AFM, unwanted mechanical contact of the AFM tip with the substrate was prevented. Precise control of the tip-substrate gap distance with FM-AFM and a gold tip sharpened with focused ion beam enable us to deposit gold dots as small as approximately 20 nm in size and construct nanoscale patterns. PMID- 19123569 TI - Microbeam-integrated multiphoton imaging system. AB - Multiphoton microscopy has been added to the array of imaging techniques at the endstation for the Microbeam II cell irradiator at Columbia University's Radiological Research Accelerator Facility (RARAF). This three-dimensional (3D), laser-scanning microscope functions through multiphoton excitation, providing an enhanced imaging routine during radiation experiments with tissuelike samples, such as small living animals and organisms. Studies at RARAF focus on radiation effects; hence, this multiphoton microscope was designed to observe postirradiation cellular dynamics. This multiphoton microscope was custom designed into an existing Nikon Eclipse E600-FN research fluorescence microscope on the irradiation platform. Design details and biology applications using this enhanced 3D-imaging technique at RARAF are reviewed. PMID- 19123570 TI - A scanning Hall probe microscope for high resolution magnetic imaging down to 300 mK. AB - We present the design, construction, and performance of a low-temperature scanning Hall probe microscope with submicron lateral resolution and a large scanning range. The detachable microscope head is mounted on the cold flange of a commercial (3)He-refrigerator (Oxford Instruments, Heliox VT-50) and operates between room temperature and 300 mK. It is fitted with a three-axis slip-stick nanopositioner that enables precise in situ adjustment of the probe location within a 6x6x7 mm(3) space. The local magnetic induction at the sample surface is mapped with an easily changeable microfabricated Hall probe [typically GsAs/AlGaAs or AlGaAs/InGaAs/GaAs Hall sensors with integrated scanning tunnel microscopy (STM) tunneling tips] and can achieve minimum detectable fields >or=10 mG/Hz(1/2). The Hall probe is brought into very close proximity to the sample surface by sensing and controlling tunnel currents at the integrated STM tip. The instrument is capable of simultaneous tunneling and Hall signal acquisition in surface-tracking mode. We illustrate the potential of the system with images of superconducting vortices at the surface of a Nb thin film down to 372 mK, and also of labyrinth magnetic-domain patterns of an yttrium iron garnet film captured at room temperature. PMID- 19123571 TI - Direct calibration of colloidal probe cantilevers via Derjaguin, Landau, Verwey, and Overbeek surface forces in electrolyte solution. AB - The development of colloidal probe microscopy has made it possible to directly measure the interaction forces between two different surfaces in solution. Cantilever calibration is presently a subject of intense experimental and theoretical interest due to the need for accurate force measurement. We developed a novel and direct calibration method for colloidal probe cantilevers to which a silica microsphere has been previously attached based on fitting experimental force curves for the interaction between the silica sphere and a silica flat in dilute KBr solutions to the theoretical Derjaguin, Landau, Verwey, and Overbeek force curves using the measured zeta potential of the silica surfaces. PMID- 19123572 TI - Development of a scanning surface probe for nanoscale tip-enhanced desorption/ablation. AB - We report on the development of a versatile scanning apparatus for nanoscale surface sampling that utilizes the interaction of laser radiation at a sharp probe tip to effect desorption/ablation on opaque substrates. The process, which currently yields surface craters as small as approximately 50 nm diameterx5 nm deep, has been demonstrated with both metal-coated and bare silicon tips. Desorption/ablation under the tip occurs at illumination intensities below the corresponding optical far-field threshold, suggesting that the latter process should not degrade the spatial resolution attainable for proposed chemical imaging methods based on the scanning surface probe. PMID- 19123573 TI - Radio-frequency reflectometry on large gated two-dimensional systems. AB - We have embedded an AlGaAs/GaAs based, gated two-dimensional (2D) hole system (2DHS) into an impedance transformer LC circuit and show that by using radio frequency reflectometry it is possible to perform sensitive, large bandwidth, electrical resistance measurements of 2D systems at millikelvin temperatures. We construct a simple lumped element model where the gated 2DHS is described as a resistive transmission line. The model gives a qualitative understanding of the experimental results. As an example, we use our method to map out the Landau level evolution in a 2DHS as a function of magnetic field and gate voltage. PMID- 19123574 TI - Cryogenic current-in-plane tunneling apparatus. AB - We have designed and fabricated a cryogenic variable-temperature current-in-plane tunneling apparatus to measure the magnetoresistive properties of unpatterned magnetic tunnel junction wafers as a function of temperature. The wafer is mounted on the cold finger of a liquid helium continuous flow cryostat. The temperature can be continuously varied between 7 and 330 K. We describe the design and fabrication of the micromachined silicon probe head that comprises a comb of 20 measuring and 4 leveling probes. The measuring probes are typically 0.7 microm wide and 1.2 microm thick, with lengths of 10, 7, and 4 microm, and a pitch that varies from 1.5 to 30 microm. The leveling probes are used in conjunction with a tilt stage to adjust the parallelism between the comb and the sample wafer during the approach of the probe head. The probe head is mounted on a nonmagnetic x-y stage, which can access a 22x22 mm(2) area with a repeatability of approximately 1 microm. The first measurements taken at room and cryogenic temperatures are shown. PMID- 19123575 TI - Semicircular bend testing with split Hopkinson pressure bar for measuring dynamic tensile strength of brittle solids. AB - We propose and validate an indirect tensile testing method to measure the dynamic tensile strength of rocks and other brittle solids: semicircular bend (SCB) testing with a modified split Hopkinson pressure bar (SHPB) system. A strain gauge is mounted near the failure spot on the specimen to determine the rupture time. The momentum trap technique is utilized to ensure single pulse loading for postmortem examination. Tests without and with pulse shaping are conducted on rock specimens. The evolution of tensile stress at the failure spot is determined via dynamic and quasistatic finite element analyses with the dynamic loads measured from SHPB as inputs. Given properly shaped incident pulse, far-field dynamic force balance is achieved and the peak of the loading matches in time with the rupture onset of the specimen. In addition, the dynamic tensile stress history at the failure spot obtained from the full dynamic finite element analysis agrees with the quasistatic analysis. The opposite occurs for the test without pulse shaping. These results demonstrate that when the far-field dynamic force balance is satisfied, the inertial effects associated with stress wave loading are minimized and thus one can apply the simple quasistatic analysis to obtain the tensile strength in the SCB-SHPB testing. This method provides a useful and cost effective way to measure indirectly the dynamic tensile strength of rocks and other brittle materials. PMID- 19123576 TI - Compact cryogenic Kerr microscope for time-resolved studies of electron spin transport in microstructures. AB - A compact cryogenic Kerr microscope for operation in the small volume of high field magnets is described. It is suited for measurements both in Voigt and Faraday configurations. Coupled with a pulsed laser source, the microscope is used to measure the time-resolved Kerr rotation response of semiconductor microstructures with approximately 1 mum spatial resolution. The microscope was designed to study spin transport, a critical issue in the field of spintronics. It is thus possible to generate spin polarization at a given location on a microstructure and probe it at a different location. The operation of the microscope is demonstrated by time-resolved measurements of micrometer distance diffusion of spin polarized electrons in a GaAs/AlGaAs heterojunction quantum well at 4.2 K and 7 T. PMID- 19123577 TI - Benchtop time-resolved magneto-optical Kerr magnetometer. AB - We present here the construction and application of a compact benchtop time resolved Kerr magnetometer to measure the magnetization precession in magnetic thin films and lithographically patterned elements. As opposed to very expensive femtosecond lasers this system is built upon a picosecond pulsed injection diode laser and electronic pulse and delay generators. The precession is triggered by the electronic pulses of controlled duration and shape, which is launched onto the sample by a microstrip line. We used polarized optical pulses synchronous to the electronic pulses to measure the magneto-optical Kerr rotation. The system is integrated in a conventional upright microscope configuration with separate illumination, imaging, and magneto-optical probe paths. The system offers high stability, relative ease of alignment, sample changing, and a long range of time delay. We demonstrate the measurements of time-resolved dynamics of a Permalloy microwire and microdot using this system, which showed dynamics at two different time scales. PMID- 19123578 TI - Heat-capacity measurements on small samples: the hybrid method. AB - A newly developed method is presented for measuring heat capacities on small samples, particularly where thermal isolation is not sufficient for the use of the traditional semiadiabatic heat-pulse technique. This "hybrid technique" is a modification of this heat-pulse method in case the temperature drift of the sample after the heat pulse is not linear but exponential. Deliberate extrapolations of these exponential drift curves, dependent on the evaluated relaxation time, yield reliable results for the temperature steps. The method is faster than the traditional relaxation method, and by comparing with the values published by the National Bureau of Standards (NBS) on copper, the accuracy is shown to be of the order of 1%. PMID- 19123579 TI - A method for removing surfactants from an air/water interface. AB - The study of heat, mass, and momentum transport across an air/water interface is an aspect of fluid mechanics where the presence of surfactant monolayers can play a significant role. Experimental studies of air/water transport typically require a method for cleaning the air/water interface so that it is free from any contaminating surfactant monolayer. This may be for the sake of running an experiment under clean surface conditions, or to clean the surface prior to deposition of a known surfactant. Herein a method is described for maintaining a clean air/water interface during conditions of finite air flow over the water surface. The unique aspect of this method is its ability to maintain clean surfaces while experiments are conducted. PMID- 19123580 TI - Tunable VUV photochemistry using Rydberg H-atom time-of-flight spectroscopy. AB - In this article, we report an experimental method for studying tunable vacuum ultraviolet (VUV) photochemistry using the H-atom Rydberg tagging technique. In this method, two VUV laser light beams were generated using nonlinear four-wave mixing scheme in a single Kr gas cell: one VUV beam is fixed at the 121.6 nm wavelength to probe the H-atom product through the Lyman alpha transition, the other beam can be tunable for photodissociating molecules in the wavelength range lambda(VUV)=121-190 nm. Preliminary results on the H(2)O photodissociation in the B state are reported here. These results suggest that the experimental method is a powerful tool for investigating photodissociation dynamics in the VUV region for molecules involving H-atom processes. PMID- 19123581 TI - Infrared spectroscopic methods for the study of aerosol particles using White cell optics: Development and characterization of a new aerosol flow tube. AB - A description of a new aerosol flow tube apparatus for measurements in situ under atmospherically relevant conditions is presented here. The system consists of a laboratory-made nebulizer generation system and a flow tube with a White cell based Fourier transform IR for the detection system. An assessment of the White cell coupled to the flow tube was carried out by an extensive set of experiments to ensure the alignment of the infrared beam and optimize the performance of this system. The detection limit for CO was established as (1.0+/-0.3) ppm and 16 passes was chosen as the optimum number of passes to be used in flow tube experiments. Infrared spectroscopy was used to characterize dry aerosol particles in the flow tube. Pure particles composed of ammonium sulfate or sodium chloride ranging between 0.8 and 2.1 mum for size diameter and (0.8-4.9)x10(6) particles/cm(3) for density number were generated by nebulization of aqueous solutions. Direct measurements of the aerosol particle size agree with size spectra retrieved from inversion of the extinction measurements using Mie calculations, where the difference residual value is in the order of 0.2%. The infrared detection limit for ammonium sulfate aerosol particles was determined as d(p)=0.9 mum and N=5x10(3) particles/cm(3) with sigma=1.1 by Mie calculation. Alternatively, Mie calculations were performed to determine the flexibility in varying the optical length when aerosol particles are sent by the injector. The very good agreement between the values retrieved for aerosol particles injected through the flow tube or through the injector clearly validates the estimation of the effective optical path length for the injector. To determine the flexibility in varying the reaction zone length, analysis of the extinction spectra as function of the position of the injector was carried out by monitoring the integrated area of different absorption modes of the ammonium sulfate. We conclude that the aerosol loss in the flow tube reactor is negligible and that the aerosol particles remain on-axis for the length of the flow tube. PMID- 19123582 TI - Fabrication of gravity-driven microfluidic device. AB - We have studied the micro total analysis system as a blood test. A microfluidic device with a three-pronged microchannel and artificial capillary vessels was fabricated. The microchannel is to transport blood, focus blood cells, and line them up. The vessels are to observe red blood cell deformation. An excimer laser was used to form grooves and so on. Numbers of thermosetting resin film and fluororesin were piled up on a cover glass. A laser fabricated part of the channel at the each film every lamination, and then a three-dimensional structure microchannel was fabricated. The channel sizes have widths of 50-150 microm and depths of 45 mum. Through holes used as artificial capillary vessels are made in the fluororesin having a minimum diameter of 5 microm and a length of 100 microm. As blood and a physiological saline are injected into the microchannel, the device stands upward facing the channel, and blood cells go into the vessels by the force of gravity and sheath flow of the saline. By gravity various groove patterns were made changing the width and length for measurement of blood focusing. Moreover, the red blood cell deformation was observed in the vessels with a microscope. PMID- 19123583 TI - Visual observation and quantitative measurement of the microwave absorbing effect at X band. AB - We have set up a two-dimensional spatial field mapping system to measure graphically the quasi-free-space electromagnetic wave in a parallel plate waveguide. Our apparatus illustrates a potential application in characterizing the microwave absorbing materials. From the measured mappings of the microwave field, the visualization of spatial physical process and quantitative characterization of reflectivity coefficients can be achieved. This simple apparatus has a remarkable advantage over with conventional testing methods which usually involve huge, expensive anechoic chambers and demand samples of large size. PMID- 19123584 TI - Fast-charging compact seed source for magnetic flux compression generators. AB - Flux compression generators (FCGs) are some of the most attractive sources of single-use compact pulsed power available today due to their high energy density output and mobility. Driving FCGs requires some seed energy, which is typically provided by applying a high seed current, usually in the kiloampere range for midsized helical FCGs. This initial current is supplied by a high-current seed source that is capable of driving an inductive load. High-current seed sources have typically been comprised of discharging large capacitors using spark gaps and overvoltage triggering mechanisms to provide the prime power for FCGs. This paper will discuss a recent design of a self-contained (battery powered with full charge time less than 40 s), single-use compact seed source (CSS) using solid state components for the switching scheme. The CSS developed is a system (0.005 m(3) volume and weighing 3.9 kg) capable of delivering over 360 J ( approximately 12 kA) into a 5.20 muH load with a trigger energy of microjoules at the TTL triggering level. The newly designed solid-state switching scheme of the CSS incorporates off-the-shelf high-voltage semiconductor components that minimize system cost and size as necessary for a single-use application. A detailed evaluation of the CSS is presented primarily focusing on the switching mechanics and experimental characterization of the solid-state components used in the system. PMID- 19123585 TI - Efficient wavelet ridge extraction method for asymptotic signal analysis. AB - A wavelet ridge extraction method is proposed for efficient estimation of instantaneous frequency and amplitude of the asymptotic signal. Inequalities are derived to provide limitations on scale interval and sampling frequency. The evaluation of optimal scale interval and sampling frequency at any given accuracy can be done by solving these inequalities and minimizing the computational complexity of continuous wavelet transform. After the continuous wavelet transform of the signal is completed with these optimal parameters, a more reliable ridge can be formed using only the discontinuity points on the ridge extracted using the conventional method. The results of a system identification simulation test performed to verify the efficiency of the proposed method indicate that the computational load of the proposed method is about 1/6 of that of the conventional method. When the system identification method is used to measure samples with known moment of inertia, the relative error below 0.08% is achieved. PMID- 19123586 TI - A sensitive and high dynamic range cw laser power meter. AB - We report the design of a cost effective, highly sensitive cw laser power meter with a large dynamic range based on a photodiode. The power meter consists of a photodiode, a current to voltage converter circuit, an offset balancing circuit, a microcontroller, an analog to digital converter, reed relays, and an alphanumeric liquid crystal display. The power meter can record absolute laser power levels as low as 1 pW. The dynamic range measured with a cw laser at a wavelength of 532 nm is 8x10(10). The high sensitivity and large dynamic range are achieved by the implementation of an analog background balancing circuit and autoranging. PMID- 19123587 TI - A nanotube based electron microbeam cellular irradiator for radiobiology research. AB - A prototype cellular irradiator utilizing a carbon nanotube (CNT) based field emission electron source has been developed for microscopic image-guided cellular region irradiation. The CNT cellular irradiation system has shown great potential to be a high temporal and spatial resolution research tool to enable researchers to gain a better understanding of the intricate cellular and intercellular microprocesses occurring following radiation deposition, which is essential to improving radiotherapy cancer treatment outcomes. In this paper, initial results of the system development are reported. The relationship between field emission current, the dose rate, and the dose distribution has been investigated. A beam size of 23 mum has been achieved with variable dose rates of 1-100 Gy/s, and the system dosimetry has been measured using a radiochromic film. Cell irradiation has been demonstrated by the visualization of H2AX phosphorylation at DNA double strand break sites following irradiation in a rat fibroblast cell monolayer. The prototype single beam cellular irradiator is a preliminary step to a multipixel cell irradiator that is under development. PMID- 19123588 TI - Resistivity and thermopower measurement setups in the temperature range of 5-325 K. AB - Automated precision measurement setups for electrical resistivity of eight metallic samples simultaneously and thermoelectric power of different types of samples in the temperature range of 5-325 K have been developed. The details of the setups and their capabilities have been described. Usually each run takes nearly 5 h and typical error is within 3% and 4%, respectively. The results of high purity Nb and Pt samples are used as examples to demonstrate this. PMID- 19123589 TI - Polarimetric interferometer for nanoscale positioning applications. AB - We propose and demonstrate a displacement control method at the subnanometric scale based on a Michelson interferometer combined with a polarimeter and a phase locked loop electronic board. Step by step displacements with a step value of 5 nm are presented. A repeatability of 0.47 nm is obtained from back and forth displacements over 1 mum range. We show that a residual ellipticity of less than 10 degrees on the polarization state leads to a positioning error of less than 1 nm. Such system could be used over millimeter range displacements in a controlled surrounding environment leading to numerous applications in nanometrology. PMID- 19123590 TI - Bubble velocity, diameter, and void fraction measurements in a multiphase flow using fiber optic reflectometer. AB - A fiber optic reflectometer (FOR) technique featuring a single fiber probe is investigated for its feasibility of measuring the bubble velocity, diameter, and void fraction in a multiphase flow. The method is based on the interference of the scattered signal from the bubble surface with the Fresnel reflection signal from the tip of the optical fiber. Void fraction is obtained with a high accuracy if an appropriate correction is applied to compensate the underestimated measurement value. Velocity information is accurately obtained from the reflected signals before the fiber tip touches the bubble surface so that several factors affecting the traditional dual-tip probes such as blinding, crawling, and drifting effects due to the interaction between the probe and bubbles can be prevented. The coherent signals reflected from both the front and rear ends of a bubble can provide velocity information. Deceleration of rising bubbles and particles due to the presence of the fiber probe is observed when they are very close to the fiber tip. With the residence time obtained, the bubble chord length can be determined by analyzing the coherent signal for velocity determination before the deceleration starts. The bubble diameters are directly obtained from analyzing the signals of the bubbles that contain velocity information. The chord lengths of these bubbles measured by FOR represent the bubble diameters when the bubble shape is spherical or represent the minor axes when the bubble shape is ellipsoidal. The velocity and size of bubbles obtained from the FOR measurements are compared with those obtained simultaneously using a high speed camera. PMID- 19123591 TI - Design of a double diaphragm shock tube for fluid disintegration studies. AB - A double diaphragm shock tube facility for studying liquid-spray atomization and combustion-related phenomena at elevated pressures and temperatures is described. The present shock tube is specifically intended for the investigation of fundamental processes related to fluid disintegration and mixing under realistic engine conditions. Special features of the facility include a variable-area driver section to compensate for shock attenuation, a square test section to allow flow visualization in the postshock region, a skimmer to dispose part of the boundary layer, a heated, fast-response injector, a fully automated gas filling system, and a new control system and electronics. Test times of the order of 2-5 ms are possible with reflected shock pressures up to 50 bar and temperatures of 2000 K. Details on the setup design, construction and operation are given. Particular emphasis is placed on the accuracy and reproducibility of the test conditions. To that aim, qualification tests have been performed to assess the shock tube performance in terms of effectiveness of the skimmer concept, the capability to compensate for boundary layer effects and the generation of uniform and reproducible test and injection conditions. PMID- 19123592 TI - A furnace to 1200 K for in situ heating x-ray diffraction, small angle x-ray scattering, and x-ray absorption fine structure experiments. AB - A furnace with a water-cooled outside shell has been assembled to do in situ x ray diffraction (XRD), small angle x-ray scattering (SAXS), and x-ray absorption fine structure (XAFS) experiments. The details of the furnace are described in this paper. The in situ XRD, SAXS, and XAFS experiments during the heating process demonstrate that the available temperature range of this furnace is from room temperature to 1200 K with a temperature accuracy of +/-0.1 K. By using this furnace, in situ XRD, SAXS, and XAFS experimental techniques with temperature change can be easily combined together. PMID- 19123593 TI - A robust floating nanoammeter. AB - A circuit capable of measuring nanoampere currents while floating at voltages up to at least 25kV is described. The circuit relays its output to ground potential via an optical fiber. We particularly emphasize the design and construction techniques, which allow robust operation in the presence of high voltage spikes and discharges. PMID- 19123594 TI - Universal serial bus powered and controlled isolated constant-current physiological stimulator. AB - We have developed a compact, isolated, physiological, constant-current stimulator that is powered and controlled by a universal serial bus (USB) interface. The stimulator is designed to be used in ex vivo cardiac experiments but is suitable for a wide variety of settings. The cost and features compare very favorably with commercial stimulators usually used in research and student laboratories. In addition to being USB powered, other novel aspects of our stimulator include the ability to produce large currents, up to 100 mA through a typical 1 kOmega load, by means of a single high-voltage dc-to-dc converter; user-specified variable period, magnitude, and duration of complex monophasic or biphasic sequences; and easy integration via hardware or software into existing experimental setups. PMID- 19123595 TI - Polarization orthogonalizer for a pair of laser beams with nearly equal frequencies. AB - We have demonstrated a device to produce an overlapping pair of orthogonally polarized laser beams with a 3 GHz frequency offset out of a single laser beam containing the two frequency components with the same linear polarization. Our design is based on a Michelson interferometer formed by a polarizing beam splitter and two quarter-waveplates. Such a device can be used to make the polarization states of a carrier and a sideband produced through modulation mutually orthogonal. An orthogonally polarized pair of coherent laser beams can be used for an interferometric measurement of a small displacement in a heterodyne scheme or to produce a large-contrast coherent population trapping signal from alkali metal atoms. As a demonstration we used the device to achieve 40% contrast for a coherent population trapping signal from a rubidium vapor cell. PMID- 19123596 TI - Low-frequency mechanical spectroscopy study of conformational transition of polymer chains in concentrated solutions. AB - A low-frequency mechanical spectroscopy approach for liquids was proposed for studying conformational transition of polymer chains in concentrated solutions. The technique is applied to aqueous solutions of a poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer in order to determine if chain conformation is altered in response to temperature. Two transitions are revealed by mechanical spectra and verified by differential thermal analysis with increasing temperature, which may be related to the unimer to-micelle transition and the phase separation, respectively. The transitions are also found to be much dependent on the concentration of the solution and the addition of NaCl. Moreover, it reveals that the PEO blocks play a more important role in the micellar crystallization process. This study may be helpful in understanding the dynamics of polymer chains in concentrated solutions. PMID- 19123597 TI - Cutting down the forest of peaks in acoustic dynamic atomic force microscopy in liquid. AB - Acoustic dynamic force microscopy in liquids is a fundamental technique for the investigation of biological samples under physiological conditions. However, it shows an important drawback that consists of producing a myriad of resonance peaks, known as the forest of peaks, which hides the natural resonance frequency of the cantilever and prevents an optimum operation of the microscope. In this work, we propose a simple remedy for this problem, which consists on adding a small clay damper to the dither piezoelectric. The resulting frequency spectrum exhibits a single resonance peak that is comparable with the one obtained using magnetic excitation. PMID- 19123598 TI - A new tensile stage for in situ electron microscopy examination of the mechanical properties of "superelastic" specimens. AB - We have developed a novel tensile stage that can be used for in situ electron microscopy examination of the mechanical properties of "superelastic" materials. In our stage, one of the specimen clamps is replaced by a cylindrical roller, which when driven by a motor can easily stretch ("roll on") any specimen irrespective of its plastic properties. We have used the so-called Roll-o-meter in the study of the tensile behavior of two different film formed latex formulations, here referred to as standard and novel. We find that the values of the tensile strength and extension to break of the studied systems, measured by using the Roll-o-meter, are similar to those measured by a Hounsfield tensile testing machine outside the microscope chamber. Further, in situ environmental scanning electron microscopy examination of the deformation and failure of the lattices revealed that the standard specimens exhibit a more ductile behavior, compared to the novel ones. PMID- 19123599 TI - Measurement of shock wave unsteadiness using a high-speed schlieren system and digital image processing. AB - A new method to measure shock wave unsteadiness is presented. Time-resolved visualizations of the flow field under investigation are obtained using a high speed schlieren optical system and the motion of the shock wave is determined by means of digital image processing. Information on the shock's unsteadiness is subsequently derived with Fourier analysis. A sample study on shock unsteadiness in a shock-wave/turbulent boundary-layer interaction with separation is included. The method presented enables a measure of shock unsteadiness at locations in the imaged flow field not accessible by intrusive methods. PMID- 19123602 TI - Detecting topological features of chaotic fluid flow. PMID- 19123603 TI - Enstrophy amplification events in three-dimensional turbulence. PMID- 19123604 TI - From bouncing to boxing. PMID- 19123605 TI - Channel erosion due to subsurface flow. PMID- 19123606 TI - Swimming C. elegans in a wet granular medium. PMID- 19123607 TI - Force chains in a two-dimensional granular pure shear experiment. PMID- 19123608 TI - Cracking sheets: oscillatory fracture paths in thin elastic sheets. PMID- 19123609 TI - A cascade of length scales in elastic rings under confinement. PMID- 19123610 TI - Momentum distributions after double ionization. PMID- 19123611 TI - Inferring connectivity of interacting phase oscillators. AB - The question as to how network topology properties influence network dynamical behavior has been extensively investigated. Here we treat the inverse problem, i.e., how to infer network connection topology from the dynamic evolution, and suggest a control based topology identification method. This method includes two steps: (i) driving the network to a steady state and (ii) inferring all elements of the connectivity matrix by exploiting information obtained from the observed steady state response of each node. We adopt different strategies for model dependent (i.e., each local phase dynamics and coupling functions are known) and model-free (i.e., each local phase dynamics and coupling functions are unknown) cases and give detailed conditions for both cases under which network topology can be identified correctly. The influence of noise on topology identification is discussed as well. All proposed approaches are motivated and illustrated with networks of phase oscillators. We argue that these topology identification methods can be extended to general dynamical networks and are not restricted to only networks of phase oscillators. PMID- 19123613 TI - Winnerless competition principle and prediction of the transient dynamics in a Lotka-Volterra model. AB - Predicting the evolution of multispecies ecological systems is an intriguing problem. A sufficiently complex model with the necessary predicting power requires solutions that are structurally stable. Small variations of the system parameters should not qualitatively perturb its solutions. When one is interested in just asymptotic results of evolution (as time goes to infinity), then the problem has a straightforward mathematical image involving simple attractors (fixed points or limit cycles) of a dynamical system. However, for an accurate prediction of evolution, the analysis of transient solutions is critical. In this paper, in the framework of the traditional Lotka-Volterra model (generalized in some sense), we show that the transient solution representing multispecies sequential competition can be reproducible and predictable with high probability. PMID- 19123614 TI - Hopf bifurcation control in a congestion control model via dynamic delayed feedback. AB - A typical objective of bifurcation control is to delay the onset of undesirable bifurcation. In this paper, the problem of Hopf bifurcation control in a second order congestion control model is considered. In particular, a suitable Hopf bifurcation is created at a desired location with preferred properties and a dynamic delayed feedback controller is developed for the creation of the Hopf bifurcation. With this controller, one can increase the critical value of the communication delay, and thus guarantee a stationary data sending rate for larger delay. Furthermore, explicit formulae to determine the period and the direction of periodic solutions bifurcating from the equilibrium are obtained by applying perturbation approach. Finally, numerical simulation results are presented to show that the dynamic delayed feedback controller is efficient in controlling Hopf bifurcation. PMID- 19123612 TI - Asymmetric spatiotemporal chaos induced by a polypoid mass in the excised larynx. AB - In this paper, asymmetric spatiotemporal chaos induced by a polypoid mass simulating the laryngeal pathology of a vocal polyp is experimentally observed using high-speed imaging in an excised larynx. Spatiotemporal analysis reveals that the normal vocal folds show spatiotemporal correlation and symmetry. Normal vocal fold vibrations are dominated mainly by the first vibratory eigenmode. However, pathological vocal folds with a polypoid mass show broken symmetry and spatiotemporal irregularity. The spatial correlation is decreased. The pathological vocal folds spread vibratory energy across a large number of eigenmodes and induce asymmetric spatiotemporal chaos. High-order eigenmodes show complicated dynamics. Spatiotemporal analysis provides a valuable biomedical application for investigating the spatiotemporal chaotic dynamics of pathological vocal fold systems with a polypoid mass and may represent a valuable clinical tool for the detection of laryngeal mass lesion using high-speed imaging. PMID- 19123615 TI - Community structures and role detection in music networks. AB - We analyze the existence of community structures in two different social networks using data obtained from similarity and collaborative features between musical artists. Our analysis reveals some characteristic organizational patterns and provides information about the driving forces behind the growth of the networks. In the similarity network, we find a strong correlation between clusters of artists and musical genres. On the other hand, the collaboration network shows two different kinds of communities: rather small structures related to music bands and geographic zones, and much bigger communities built upon collaborative clusters with a high number of participants related through the period the artists were active. Finally, we detect the leading artists inside their corresponding communities and analyze their roles in the network by looking at a few topological properties of the nodes. PMID- 19123616 TI - The organization of intrinsic computation: complexity-entropy diagrams and the diversity of natural information processing. AB - Intrinsic computation refers to how dynamical systems store, structure, and transform historical and spatial information. By graphing a measure of structural complexity against a measure of randomness, complexity-entropy diagrams display the different kinds of intrinsic computation across an entire class of systems. Here, we use complexity-entropy diagrams to analyze intrinsic computation in a broad array of deterministic nonlinear and linear stochastic processes, including maps of the interval, cellular automata, and Ising spin systems in one and two dimensions, Markov chains, and probabilistic minimal finite-state machines. Since complexity-entropy diagrams are a function only of observed configurations, they can be used to compare systems without reference to system coordinates or parameters. It has been known for some time that in special cases complexity entropy diagrams reveal that high degrees of information processing are associated with phase transitions in the underlying process space, the so-called "edge of chaos." Generally, though, complexity-entropy diagrams differ substantially in character, demonstrating a genuine diversity of distinct kinds of intrinsic computation. PMID- 19123617 TI - Multistability and arithmetically period-adding bifurcations in piecewise smooth dynamical systems. AB - Multistability has been a phenomenon of continuous interest in nonlinear dynamics. Most existing works so far have focused on smooth dynamical systems. Motivated by the fact that nonsmooth dynamical systems can arise commonly in realistic physical and engineering applications such as impact oscillators and switching electronic circuits, we investigate multistability in such systems. In particular, we consider a generic class of piecewise smooth dynamical systems expressed in normal form but representative of nonsmooth systems in realistic situations, and focus on the weakly dissipative regime and the Hamiltonian limit. We find that, as the Hamiltonian limit is approached, periodic attractors can be generated through a series of saddle-node bifurcations. A striking phenomenon is that the periods of the newly created attractors follow an arithmetic sequence. This has no counterpart in smooth dynamical systems. We provide physical analyses, numerical computations, and rigorous mathematical arguments to substantiate the finding. PMID- 19123618 TI - Dynamical parameter identification from a scalar time series. AB - If a drive system with unknown parameters represents "reality" and the response system a "computational model," the unidirectional coupling can be used to change model parameters, as well as the model state, such that both systems synchronize with each other and model parameters coincide with their true values of "reality." Such a parameter identification method is called adaptive synchronization (also autosynchronization) method and is widely used in the literature. Because one usually cannot find proper parameter update rules by exploiting information obtained from only a scalar time series, parameter identification with adaptive synchronization from a scalar time series is not well understood and still remains challenging until now. In this paper we introduce a novel adaptive synchronization approach with an effective guidance parameter to update rule design. This method includes three steps: (i) finding some proper control signals such that the "computational model" synchronizes with the "real" system if no parameter mismatch exists (that is, both systems have identical parameters); (ii) designing parameter update rules in terms of a necessary condition for ensuring local synchronization; and (iii) determining the value for each parameter update rate for ensuring the local stability of autosynchronization manifold according to the conditional Lyapunov exponents method. The reliability of the suggested technique is illustrated with the Lorenz system and a unified chaotic model. PMID- 19123619 TI - Synchronization-based scalability of complex clustered networks. AB - Complex clustered networks arise in biological, social, physical, and technological systems, and the synchronous dynamics on such networks have attracted recent interests. Here we investigate system-size dependence of the synchronizability of these networks. Theoretical analysis and numerical computations reveal that, for a typical clustered network, as its size is increased, the synchronizability can be maintained or even enhanced but at the expense of deterioration of the clustered characteristics in the topology that distinguish this type of networks from other types of complex networks. An implication is that, for a large network in a realistic situation, if synchronization is important for its function, then most likely it will not have a clustered topology. PMID- 19123620 TI - Cryptanalysis of a chaotic communication scheme using adaptive observer. AB - This paper addresses the cryptanalysis of a secure communication scheme recently proposed by Wu [Chaos 16, 043118 (2006)], where the information signal is modulated into a system parameter of a unified chaotic system. With the Kerckhoff principle, assuming that the structure of the cryptosystem is known, an adaptive observer can be designed to synchronize the targeted system, so that the transmitted information and the user-specific parameters are obtained. The success of adaptive synchronization is mathematically proved with the use of Lyapunov stability theory, based on the original assumption, i.e., the dynamical evolution of the information signal is available. A more practical case, but yet much more difficult, is also considered. As demonstrated with simulations, generalized synchronization is still possible, even if the derivative of the information signal is kept secret. Hence, the message can be coarsely estimated, making the security of the considered system questionable. PMID- 19123622 TI - Synchronization of a class of chaotic systems with fully unknown parameters using adaptive sliding mode approach. AB - In this paper, an adaptive sliding mode control method for synchronization of a class of chaotic systems with fully unknown parameters is introduced. In this method, no knowledge of the bounds of parameters is required in advance and the parameters are updated through an adaptive control process. We use our proposed method to synchronize two chaotic gyros, which has been the subject of intense study during the recent years for its application in the navigational, aeronautical, and space engineering domains. The effectiveness of our method is demonstrated in simulation environment and the results are compared with some recent schemes proposed in the literature for the same task. PMID- 19123621 TI - Pinning synchronization of delayed neural networks. AB - This paper investigates adaptive pinning synchronization of a general weighted neural network with coupling delay. Unlike recent works on pinning synchronization which proposed the possibility that synchronization can be reached by controlling only a small fraction of neurons, this paper aims to answer the following question: Which neurons should be controlled to synchronize a neural network? By using Schur complement and Lyapunov function methods, it is proved that under a mild topology-based condition, some simple adaptive feedback controllers are sufficient to globally synchronize a general delayed neural network. Moreover, for a concrete neurobiological network consisting of identical Hindmarsh-Rose neurons, a specific pinning control technique is introduced and some numerical examples are presented to verify our theoretical results. PMID- 19123623 TI - Stability and multiple bifurcations of a damped harmonic oscillator with delayed feedback near zero eigenvalue singularity. AB - We investigate the dynamics of a damped harmonic oscillator with delayed feedback near zero eigenvalue singularity. We perform a linearized stability analysis and multiple bifurcations of the zero solution of the system near zero eigenvalue singularity. Taking the time delay as the bifurcation parameter, the presence of steady-state bifurcation, Bogdanov-Takens bifurcation, triple zero, and Hopf-zero singularities is demonstrated. In the case when the system has a simple zero eigenvalue, center manifold reduction and normal form theory are used to investigate the stability and the types of steady-state bifurcation. The stability of the zero solution of the system near the simple zero eigenvalue singularity is completely solved. PMID- 19123624 TI - On the bifurcation of species. AB - We propose and analyze a model of evolution of species based upon a general description of phenotypes in terms of a single quantifiable characteristic. In the model, species spontaneously arise as solitary waves whose members almost never mate with those in other species, according to the rules laid down. The solitary waves in the model bifurcate and we interpret such events as speciation. Our aim in this work is to determine whether a generic mathematical mechanism may be identified with this process of speciation. Indeed, there is such a process in our model: it is the Andronov homoclinic bifurcation. It is robust and is at the heart of the formation of new solitary waves, and thus (in our model) new species. PMID- 19123625 TI - Complete periodic synchronization in coupled systems. AB - Recently, complete chaotic synchronization in coupled systems has been well studied. In this paper, we study complete synchronization in coupled periodic oscillators with diffusive and gradient couplings. Eight typical types of critical curve for the transverse Lyapunov exponent of standard mode, which give rise to different synchronization-desynchronization patterns, are classified. All possible desynchronous behaviors including steady state, periodic state, quasiperiodic state, low-dimensional chaotic state, and two types of high dimensional chaotic state are identified, and two classical synchronization desynchronizaiton bifurcations--the shortest wavelength bifurcation and Hopf bifurcation from synchronous periodic state--are classified. PMID- 19123627 TI - An equivalent condition for a sequence to be the sequence of partial entropies. AB - We show that a sequence is the sequence of partial entropies of some dynamical system with respect to a partition of the phase space if and only if it is non negative, increasing, and has decreasing increments. PMID- 19123626 TI - Coherence resonance induced by colored noise near Hopf bifurcation. AB - Effects of colored noise near supercritical Hopf bifurcation, especially noise induced oscillation (NIO) and coherence resonance (CR), have been studied analytically in the Brusselator model, using the stochastic normal form method. Two types of colored noise are considered: one is the standard Gaussian colored noise generated by the Ornstein-Uhlenbeck (OU) process and the other is the so called power-limited (PL) process. Depending on the noise intensity and noise type, it is found that the autocorrelation time, most probable radius and signal to noise ratio of the NIO may show nontrivial dependencies on the noise correlation time tau(c). Interestingly, for OU-type noise with intensity above a threshold, SNR is a bell-shaped function of tau(c), indicating enhancement of CR by noise correlation; and for PL-type noise, SNR may show double maxima when tau(c) is changed, demonstrating a new kind of multiresonance phenomenon. These theoretical predictions are well reproduced by numerical simulations. PMID- 19123628 TI - Filtering by nonlinear systems. AB - Synchronization of nonlinear systems forced by external signals is formalized as the response of a nonlinear filter. Sufficient conditions for a nonlinear system to behave as a filter are given. Some examples of generalized chaos synchronization are shown to actually be special cases of nonlinear filtering. PMID- 19123629 TI - Perception-action map learning in controlled multiscroll systems applied to robot navigation. AB - In this paper a new technique for action-oriented perception in robots is presented. The paper starts from exploiting the successful implementation of the basic idea that perceptual states can be embedded into chaotic attractors whose dynamical evolution can be associated with sensorial stimuli. In this way, it can be possible to encode, into the chaotic dynamics, environment-dependent patterns. These have to be suitably linked to an action, executed by the robot, to fulfill an assigned mission. This task is addressed here: the action-oriented perception loop is closed by introducing a simple unsupervised learning stage, implemented via a bio-inspired structure based on the motor map paradigm. In this way, perceptual meanings, useful for solving a given task, can be autonomously learned, based on the environment-dependent patterns embedded into the controlled chaotic dynamics. The presented framework has been tested on a simulated robot and the performance have been successfully compared with other traditional navigation control paradigms. Moreover an implementation of the proposed architecture on a Field Programmable Gate Array is briefly outlined and preliminary experimental results on a roving robot are also reported. PMID- 19123630 TI - Improvements and applications of entrainment control for nonlinear dynamical systems. AB - This paper improves the existing entrainment control approaches and develops unified schemes to chaos control and generalized (lag, anticipated, and complete) synchronization of nonlinear dynamical systems. By introducing impulsive effects to the open-loop control method, we completely remove its restrictions on goal dynamics and initial conditions, and derive a sufficient condition to estimate the upper bound of impulsive intervals to ensure the global asymptotic stability. We then propose two effective ways to implement the entrainment strategy which combine open-loop and closed-loop control, and we prove that the feedback gains can be chosen according to a lower bound or be tuned with an adaptive control law. Numerical examples are given to verify the theoretical results and to illustrate their applications. PMID- 19123631 TI - Modulational instability in a purely nonlinear coupled complex Ginzburg-Landau equations through a nonlinear discrete transmission line. AB - We study wave propagation in a nonlinear transmission line with dissipative elements. We show analytically that the telegraphers' equations of the electrical transmission line can be modeled by a pair of continuous coupled complex Ginzburg Landau equations, coupled by purely nonlinear terms. Based on this system, we investigated both analytically and numerically the modulational instability (MI). We produce characteristics of the MI in the form of typical dependence of the instability growth rate on the wavenumbers and system parameters. Generic outcomes of the nonlinear development of the MI are investigated by dint of direct simulations of the underlying equations. We find that the initial modulated plane wave disintegrates into waves train. An apparently turbulent state takes place in the system during the propagation. PMID- 19123632 TI - Spatiotemporal system reconstruction using Fourier spectral operators and structure selection techniques. AB - A technique based on trigonometric spectral methods and structure selection is proposed for the reconstruction, from observed time series, of spatiotemporal systems governed by nonlinear partial differential equations of polynomial type with terms of arbitrary derivative order and nonlinearity degree. The system identification using Fourier spectral differentiation operators in conjunction with a structure selection procedure leads to a parsimonious model of the original system by detecting and eliminating the redundant parameters using orthogonal decomposition of the state data. Implementation of the technique is exemplified for a highly stiff reaction-diffusion system governed by the Kuramoto Sivashinsky equation. Numerical experiments demonstrate the superior performance of the proposed technique in terms of accuracy as well as robustness, even with smaller sets of sampling data. PMID- 19123633 TI - Beyond harmonic sounds in a simple model for birdsong production. AB - In this work we present an analysis of the dynamics displayed by a simple bidimensional model of labial oscillations during birdsong production. We show that the same model capable of generating tonal sounds can present, for a wide range of parameters, solutions which are spectrally rich. The role of physiologically sensible parameters is discussed in each oscillatory regime, allowing us to interpret previously reported data. PMID- 19123634 TI - The development of truncated inviscid turbulence and the Fermi-Pasta-Ulam problem. AB - A study was made of the possible similarity between the development of truncated, inviscid turbulence and the Fermi-Pasta-Ulam (FPU) problem. For the case of a constant time scale, which resembles the FPU problem, a significant increase in the time to achieve equipartition was found when the initial energy was decreased. At first a few modes were generated and only rather late in the spectral development other modes appear and equipartition was established. PMID- 19123635 TI - Phase synchronization in tilted inertial ratchets as chaotic rotators. AB - The phenomenon of phase synchronization for a particle in a periodic ratchet potential is studied. We consider the deterministic dynamics in the underdamped case where the inertia plays an important role since the dynamics can become chaotic. The ratchet potential is tilted due to a constant external force and is rocking by an external periodic forcing. This potential has to be tilted in order to obtain a rotator or self-sustained nonlinear oscillator in the absence of the external periodic forcing; this oscillator then acquires an intrinsic frequency that can be locked with the frequency of the external driving. We introduced an instantaneous linear phase, using a set of discrete time markers, and the associated average frequency, and show that this frequency can be synchronized with the frequency of the driving. We calculate Arnold tongues in a two dimensional parameter space and discuss their implications for the chaotic transport in ratchets. We show that the local maxima in the current correspond to the borders of these Arnold tongues; in this way we established a link between optimal transport in ratchets and phase synchronization. PMID- 19123636 TI - Global dissipativity analysis on uncertain neural networks with mixed time varying delays. AB - In this paper, the problems of global dissipativity and global exponential dissipativity are investigated for uncertain neural networks with discrete time varying delay and distributed time-varying delay as well as general activation functions. By constructing appropriate Lyapunov-Krasovskii functionals and employing Newton-Leibniz formulation and linear matrix inequality (LMI) technique, several new criteria for checking the global dissipativity and global exponential dissipativity of the addressed neural networks are established in terms of LMI, which can be checked numerically using the effective LMI toolbox in MATLAB. Illustrated examples are given to show the effectiveness and decreased conservatism of the proposed criteria in comparison with some existing results. It is noteworthy that the traditional assumptions on the differentiability of the time-varying delays and the boundedness of its derivative are removed. PMID- 19123637 TI - Vortex filament dynamics in computational models of ventricular fibrillation in the heart. AB - In three-dimensional cardiac tissue, the re-entrant waves that sustain ventricular fibrillation rotate around a line of phase singularity or vortex filament. The aim of this study was to investigate how the behavior of these vortex filaments is influenced by membrane kinetics, initial conditions, and tissue geometry in computational models of excitable tissue. A monodomain model of cardiac tissue was used, with kinetics described by a three-variable simplified ionic model (3V-SIM). Two versions of 3V-SIM were used, one with steep action potential duration restitution, and one with reduced excitability. Re entrant fibrillation was then simulated in three tissue geometries: a cube, a slab, and an anatomically detailed model of rabbit ventricles. Filaments were identified using a phase-based method, and the number, size, origin, and orientation of filaments was tracked throughout each simulation. The main finding of this study is that kinetics, initial conditions, geometry, and anisotropy all affected the number, proliferation, and orientation of vortex filaments in re entrant fibrillation. An important finding of this study was that the behavior of vortex filaments in simplified slab geometry representing part of the ventricular wall did not necessarily predict behavior in an anatomically detailed model of the rabbit ventricles. PMID- 19123638 TI - Stability diagram for the forced Kuramoto model. AB - We analyze the periodically forced Kuramoto model. This system consists of an infinite population of phase oscillators with random intrinsic frequencies, global sinusoidal coupling, and external sinusoidal forcing. It represents an idealization of many phenomena in physics, chemistry, and biology in which mutual synchronization competes with forced synchronization. In other words, the oscillators in the population try to synchronize with one another while also trying to lock onto an external drive. Previous work on the forced Kuramoto model uncovered two main types of attractors, called forced entrainment and mutual entrainment, but the details of the bifurcations between them were unclear. Here we present a complete bifurcation analysis of the model for a special case in which the infinite-dimensional dynamics collapse to a two-dimensional system. Exact results are obtained for the locations of Hopf, saddle-node, and Takens Bogdanov bifurcations. The resulting stability diagram bears a striking resemblance to that for the weakly nonlinear forced van der Pol oscillator. PMID- 19123639 TI - Synchronization of chaotic systems with uncertain chaotic parameters by linear coupling and pragmatical adaptive tracking. AB - We study the synchronization of general chaotic systems which satisfy the Lipschitz condition only, with uncertain chaotic parameters by linear coupling and pragmatical adaptive tracking. The uncertain parameters of a system vary with time due to aging, environment, and disturbances. A sufficient condition is given for the asymptotical stability of common zero solution of error dynamics and parameter update dynamics by the Ge-Yu-Chen pragmatical asymptotical stability theorem based on equal probability assumption. Numerical results are studied for a Lorenz system and a quantum cellular neural network oscillator to show the effectiveness of the proposed synchronization strategy. PMID- 19123640 TI - Comment on "Synchronization of chaotic systems with delay using intermittent linear state feedback" [Chaos 18, 033122 (2008)]. AB - In the referenced paper, there is technical carelessness in the third lemma and in the main result. Hence, it is a possible failure when the result is used to design the intermittent linear state feedback controller for exponential synchronization of two chaotic delayed systems. PMID- 19123642 TI - Simultaneous in vivo dynamic magnetic resonance-diffuse optical tomography for small animal imaging. AB - We present simultaneous measurement of enhancement kinetics of an optical and a magnetic resonance (MR) contrast agent in a small animal breast tumor model (R3230 ac) using a combined MR-diffuse optical tomographic (MR-DOT) imaging system. A mixture of a small molecular-weight MR contrast agent gadolinium diethylene-triamine-pentaacetic acid (Gd-DTPA) and a large molecular-weight optical contrast agent indocyanine green (ICG) was administered intravenously for multimodal dynamic imaging. Coregistration of optical and MR images was accomplished using agar-water-based markers. Using T(2) and dynamic T(1) weighted MR images, we divided the entire tumor into two regions of interest (ROI): a viable and a nonviable region. The absorption enhancements in the ROIs were calculated. An enhancement of the ICG was observed in the viable region. On the contrary, there was a lower enhancement in the nonviable region. PMID- 19123643 TI - Flexible miniature compound lens design for high-resolution optical coherence tomography balloon imaging catheter. AB - We report on a new optics design for an optical coherence tomography (OCT) balloon imaging catheter. The design involves a miniature compound gradient-index (GRIN) rod lens, which consists of a fiber optic mode-field reducer and relay rod lenses to achieve predictable high lateral resolution at a desired large working distance. The compound lens design significantly simplifies the engineering process for an OCT catheter and enables 3-D full circumferential cross sectional imaging of large luminal organs such as human esophagus. An as-designed OCT catheter is developed and demonstrated for real-time in vivo swine esophagus imaging in a 3-D spiral fashion. PMID- 19123645 TI - Parallel computing with graphics processing units for high-speed Monte Carlo simulation of photon migration. AB - General-purpose computing on graphics processing units (GPGPU) is shown to dramatically increase the speed of Monte Carlo simulations of photon migration. In a standard simulation of time-resolved photon migration in a semi-infinite geometry, the proposed methodology executed on a low-cost graphics processing unit (GPU) is a factor 1000 faster than simulation performed on a single standard processor. In addition, we address important technical aspects of GPU-based simulations of photon migration. The technique is expected to become a standard method in Monte Carlo simulations of photon migration. PMID- 19123644 TI - Polarization effect on the depth resolution of optical coherence tomography. AB - We find for the first time that polarization mismatch of the sample and reference arms in optical-fiber-based optical coherence tomography (OCT) has critical effect on its depth resolution when the light source is partially polarized. When the polarization states of the two arms are matched, the measured point spread function (PSF) is almost identical to the theoretical prediction. When their polarization states are mismatched, the PSF can be so distorted that the depth resolution is degraded to several times the theoretical value. When we polarize the source light with a polarizer, then the degree of polarization (DOP) is unity, and the depth resolution becomes independent of the polarization mismatch. This discovery has fundamental importance for high-resolution OCT imaging of biological tissues. With DOP<1, the depth resolution can be quickly degraded by either birefringence or scattering in the sample. Adjusting polarization controllers can only improve the depth resolution at a certain depth in a sample if the polarization state of light changes along the depth. When DOP=1, uniform resolution along the depth of a sample can be achieved. PMID- 19123646 TI - Cost-effective diffuse reflectance spectroscopy device for quantifying tissue absorption and scattering in vivo. AB - A hybrid optical device that uses a multimode fiber coupled to a tunable light source for illumination and a 2.4-mm photodiode for detection in contact with the tissue surface is developed as a first step toward our goal of developing a cost effective, miniature spectral imaging device to map tissue optical properties in vivo. This device coupled with an inverse Monte Carlo model of reflectance is demonstrated to accurately quantify tissue absorption and scattering in tissue like turbid synthetic phantoms with a wide range of optical properties. The overall errors for quantifying the absorption and scattering coefficients are 6.0+/-5.6 and 6.1+/-4.7%, respectively. Compared with fiber-based detection, having the detector right at the tissue surface can significantly improve light collection efficiency, thus reducing the requirement for sophisticated detectors with high sensitivity, and this design can be easily expanded into a quantitative spectral imaging system for mapping tissue optical properties in vivo. PMID- 19123647 TI - Live imaging of blood flow in mammalian embryos using Doppler swept-source optical coherence tomography. AB - Studying hemodynamic changes during early mammalian embryonic development is critical for further advances in prevention, diagnostics, and treatment of congenital cardiovascular (CV) birth defects and diseases. Doppler optical coherence tomography (OCT) has been shown to provide sensitive measurements of blood flow in avian and amphibian embryos. We combined Doppler swept-source optical coherence tomography (DSS-OCT) and live mouse embryo culture to analyze blood flow dynamics in early embryos. SS-OCT structural imaging was used for the reconstruction of embryo morphology and the orientation of blood vessels, which is required for calculating flow velocity from the Doppler measurements. Spatially and temporally resolved blood flow profiles are presented for the dorsal aorta and a yolk sac vessel in a 9.5-day embryo. We demonstrate that DSS OCT can be successfully used for structural analysis and spatially and temporally resolved hemodynamic measurements in developing early mammalian embryos. PMID- 19123648 TI - Pinhole shifting lifetime imaging microscopy. AB - Lifetime imaging microscopy is a powerful tool to probe biological phenomena independent of luminescence intensity and fluorophore concentration. We describe time-resolved imaging of long-lifetime luminescence with an unmodified commercial laser scanning confocal/multiphoton microscope. The principle of the measurement is displacement of the detection pinhole to collect delayed luminescence from a position lagging the rasting laser beam. As proof of principle, luminescence from microspheres containing europium (Eu(3+)), a red emitting probe, was compared to that of short-lifetime green-fluorescing microspheres and/or fluorescein and rhodamine in solution. Using 720-nm two-photon excitation and a pinhole diameter of 1 Airy unit, the short-lifetime fluorescence of fluorescein, rhodamine and green microspheres disappeared much more rapidly than the long-lifetime phosphorescence of Eu(3+) microspheres as the pinhole was repositioned in the lagging direction. In contrast, repositioning of the pinhole in the leading and orthogonal directions caused equal loss of short- and long-lifetime luminescence. From measurements at different lag pinhole positions, a lifetime of 270 micros was estimated for the Eu(3+) microspheres, consistent with independent measurements. This simple adaptation is the basis for quantitative 3-D lifetime imaging microscopy. PMID- 19123649 TI - Spectral filtering for improved pulsed photothermal temperature profiling in agar tissue phantoms. AB - We present a systematic experimental comparison of pulsed photothermal temperature profiling utilizing the customary spectral band of the InSb radiation detector (lambda=3.0 to 5.6 microm) and a narrowed acquisition band (4.5 to 5.6 microm). We use custom tissue phantoms composed of agar gel layers separated by thin absorbing layers. The laser-induced temperature profiles are reconstructed within the customary monochromatic approximation, using a custom minimization algorithm. In a detailed numerical simulation of the experimental procedure, we consider several acquisition spectral bands with the lower wavelength limit varied between 3.0 and 5.0 microm (imitating application of different long-pass filters). The simulated PPTR signals contain noise with amplitude and spectral characteristics consistent with our experimental system. Both experimental and numerical results indicate that spectral filtering reduces reconstruction error and broadening of temperature peaks, especially for shallower and more complex absorbing structures. For the simulated PPTR system and watery tissues, numerical results indicate an optimal lower wavelength limit of 3.8 to 4.2 microm. PMID- 19123650 TI - Retinal blood flow measurement by circumpapillary Fourier domain Doppler optical coherence tomography. AB - We present in vivo human total retinal blood flow measurements using Doppler Fourier domain optical coherence tomography (OCT). The scan pattern consisted of two concentric circles around the optic nerve head, transecting all retinal branch arteries and veins. The relative positions of each blood vessel in the two OCT conic cross sections were measured and used to determine the angle between the OCT beam and the vessel. The measured angle and the Doppler shift profile were used to compute blood flow in the blood vessel. The flows in the branch veins was summed to give the total retinal blood flow at one time point. Each measurement of total retinal blood flow was completed within 2 s and averaged. The total retinal venous flow was measured in one eye each of two volunteers. The results were 52.90+/-2.75 and 45.23+/-3.18 microlmin, respectively. Volumetric flow rate positively correlated with vessel diameter. This new technique may be useful in the diagnosis and treatment of optic nerve and retinal diseases that are associated with poor blood flow, such as glaucoma and diabetic retinopathy. PMID- 19123651 TI - Spectral discrimination of live prostate and bladder cancer cell lines using Raman optical tweezers. AB - An investigation into the use of Raman optical tweezers to study urological cell lines is reported, with the ultimate aim of determining the presence of malignant CaP cells in urine and peripheral fluids. To this end, we trapped and analyzed live CaP cells (PC-3) and bladder cells (MGH-U1), because both prostate and bladder cells are likely to be present in urine. The laser excitation wavelength of 514.5 nm was used, with Raman light collected both in back- and forward scattering geometric configurations. For the backscattering configuration the same laser was used for trapping and excitation, while for forward scattering a 1064 nm laser provided the trapping beam. Analysis of cell-diameter distributions for cells analyzed suggested normal distribution of cell sizes, indicating an unbiased cell-selection criterion. Principal components analysis afforded discrimination of MGH-U1 and PC-3 spectra collected in either configuration, demonstrating that it is possible to trap, analyze, and differentiate PC-3 from MGH-U1 cells using a 514.5 nm laser. By loading plot analysis, possible biomolecules responsible for discrimination in both configurations were determined. Finally, the effect of cell size on discrimination was investigated, with results indicating that separation is based predominantly on cell type rather than cell size. PMID- 19123652 TI - Multiscale structural analysis of mouse lingual myoarchitecture employing diffusion spectrum magnetic resonance imaging and multiphoton microscopy. AB - The tongue consists of a complex, multiscale array of myofibers that comprise the anatomical underpinning of lingual mechanical function. 3-D myoarchitecture was imaged in mouse tongues with diffusion spectrum magnetic resonance imaging (DSI) at 9.4 T (b(max) 7000 smm, 150-microm isotropic voxels), a method that derives the preferential diffusion of water/voxel, and high-throughput (10 fps) two photon microscope (TPM). Net fiber alignment was represented for each method in terms of the local maxima of an orientational distribution function (ODF) derived from the local diffusion (DSI) and 3-D structural autocorrelation (TPM), respectively. Mesoscale myofiber tracts were generated by alignment of the principal orientation vectors of the ODFs. These data revealed a consistent relationship between the properties of the respective ODFs and the virtual superimposition of the distributed mesoscale myofiber tracts. The identification of a mesoscale anatomical construct, which specifically links the microscopic and macroscopic spatial scales, provides a method for relating the orientation and distribution of cells and subcellular components with overall tissue morphology, thus contributing to the development of multiscale methods for mechanical analysis. PMID- 19123653 TI - Real-time optical monitoring of radio-frequency tissue fusion by continuous wave transmission spectroscopy. AB - Radio-frequency (RF) tissue fusion is a novel method of tissue approximation that can seal tissue without the need for sutures or staples, based on the combined effects of heat and pressure on the apposed tissue surfaces. RF delivery must be controlled and optimized to obtain a reproducible, reliable seal. We use real time optical measurements to improve understanding of the tissue modifications induced by RF fusion. The main macroscopic transformations are thermal denaturation and dehydration. Light propagation in tissue is a function of both and therefore should provide interesting insight into the dynamic of occurring phenomena. Quantification by continuous wave technique has proven challenging. We proposed an algorithm based on the measurement of the absolute transmittance of the tissue, making use of the modified Beer-Lambert law. The experimental method and the data algorithm are demonstrated by RF fusion of porcine small bowel. The proposed optical measurement modality is well adapted to modern surgical instrumentation used for minimally invasive procedures. PMID- 19123654 TI - Multimodal facial color imaging modality for objective analysis of skin lesions. AB - We introduce a multimodal facial color imaging modality that provides a conventional color image, parallel and cross-polarization color images, and a fluorescent color image. We characterize the imaging modality and describe the image analysis methods for objective evaluation of skin lesions. The parallel and cross-polarization color images are useful for the analysis of skin texture, pigmentation, and vascularity. The polarization image, which is derived from parallel and cross-polarization color images, provides morphological information of superficial skin lesions. The fluorescent color image is useful for the evaluation of skin chromophores excited by UV-A radiation. In order to demonstrate the validity of the new imaging modality in dermatology, sample images were obtained from subjects with various skin disorders and image analysis methods were applied for objective evaluation of those lesions. In conclusion, we are confident that the imaging modality and analysis methods should be useful tools to simultaneously evaluate various skin lesions in dermatology. PMID- 19123655 TI - High-speed two-photon excited autofluorescence imaging of ex vivo human retinal pigment epithelial cells toward age-related macular degeneration diagnostic. AB - Age-related macular degeneration (AMD) is among the major concerns in ophthalmology, as it is the primary cause for irreversible blindness in developed countries. Nevertheless, there is poor understanding of the origins and mechanisms that trigger this important ocular disease. In common clinical pratice, AMD is monitored by autofluorescence imaging of the retinal pigment epithelial (RPE) cells through a confocal scanning laser ophthalmoscope. The RPE cells derive their dominant autofluorescence from the lipofuscin granules that accumulate in the cytoplasm with increasing age and disease. We explored a different approach to retinal RPE imaging using two-photon excited autofluorescence, offering intrinsic three-dimensional resolution, larger sensing depth and reduced photodamage compared to single-photon excited fluorescence ophthalmoscopy. A two-photon microscope, based on the architecture of a conventional scanning laser ophthalmoscope (HRT, Heidelberg Engineering, Germany), was designed for autofluorescence imaging on retina samples from postmortem human-donor eyes. We were able to visualize at video-rate speed single RPE lipofuscin granules, demonstrating the potential to develop this method toward clinical practice for patients with RPE-related retinal disease like AMD. PMID- 19123656 TI - Application of attenuated total reflection Fourier transform infrared imaging and tape-stripping to investigate the three-dimensional distribution of exogenous chemicals and the molecular organization in Stratum corneum. AB - Attenuated total reflection Fourier transform infrared spectroscopic imaging combined with tape-stripping is an advantageous approach to map the depth penetration and lateral distribution of topically applied chemicals in Stratum corneum (SC) and the conformational order of SC lipids. Tape-stripping progressively removes layers of SC, and chemical imaging provides spatially resolved information on the chemical composition of both the newly exposed SC surface and of the tapes used for stripping. The procedure is rapid, minimally invasive, and does not necessitate cross-sectioning of the skin. This approach offers a simple and direct way to determine the distribution of exogenous volatile and non-volatile chemicals in SC as a function of the chemical composition of the formulation and time, and the conformational order of SC lipids in native and topically treated skin. The procedure described here is well suited to address questions of relevance for the areas of drug delivery, dermatology, and skin care. PMID- 19123657 TI - Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies. AB - Liver fibrosis is associated with an abnormal increase in an extracellular matrix in chronic liver diseases. Quantitative characterization of fibrillar collagen in intact tissue is essential for both fibrosis studies and clinical applications. Commonly used methods, histological staining followed by either semiquantitative or computerized image analysis, have limited sensitivity, accuracy, and operator dependent variations. The fibrillar collagen in sinusoids of normal livers could be observed through second-harmonic generation (SHG) microscopy. The two-photon excited fluorescence (TPEF) images, recorded simultaneously with SHG, clearly revealed the hepatocyte morphology. We have systematically optimized the parameters for the quantitative SHG/TPEF imaging of liver tissue and developed fully automated image analysis algorithms to extract the information of collagen changes and cell necrosis. Subtle changes in the distribution and amount of collagen and cell morphology are quantitatively characterized in SHG/TPEF images. By comparing to traditional staining, such as Masson's trichrome and Sirius red, SHG/TPEF is a sensitive quantitative tool for automated collagen characterization in liver tissue. Our system allows for enhanced detection and quantification of sinusoidal collagen fibers in fibrosis research and clinical diagnostics. PMID- 19123658 TI - Laser-induced retinal damage threshold measurements with wavefront correction. AB - An adaptive optics (AO) system was incorporated into a laser retinal exposure setup in order to correct for refractive error and higher-order aberrations of the nonhuman primate (NHP) eye during an in vivo retinal ED(50) measurement. Using this system, the ED(50) for a 100-ms, 532-nm small spot size exposure was measured to be 1.05 mJ total intraocular energy (TIE), a reduction of 22% from the value measured without aberration correction. The ED(50) for a 3.5-ns, 532-nm exposure was measured to be 0.51 microJ TIE, the lowest ED(50) reported for a ns duration exposure. This is a reduction of 37% from the value measured without aberration correction and is a factor of only 2.6 higher than the maximum permissible exposure (MPE) for a 3.5-ns, visible wavelength small spot size exposure. The trend of in vitro measurements using retinal explants suggests that the in vivo ED(50) for small spot-size exposures could potentially be one order of magnitude smaller than the previously reported in vivo ED(50). Distortion of the incident laser beam by ocular aberrations cannot fully explain the discrepancy between the in vivo measurements with no aberration correction and the in vitro results. PMID- 19123659 TI - Monte Carlo model to describe depth selective fluorescence spectra of epithelial tissue: applications for diagnosis of oral precancer. AB - We present a Monte Carlo model to predict fluorescence spectra of the oral mucosa obtained with a depth-selective fiber optic probe as a function of tissue optical properties. A model sensitivity analysis determines how variations in optical parameters associated with neoplastic development influence the intensity and shape of spectra, and elucidates the biological basis for differences in spectra from normal and premalignant oral sites. Predictions indicate that spectra of oral mucosa collected with a depth-selective probe are affected by variations in epithelial optical properties, and to a lesser extent, by changes in superficial stromal parameters, but not by changes in the optical properties of deeper stroma. The depth selective probe offers enhanced detection of epithelial fluorescence, with 90% of the detected signal originating from the epithelium and superficial stroma. Predicted depth-selective spectra are in good agreement with measured average spectra from normal and dysplastic oral sites. Changes in parameters associated with dysplastic progression lead to a decreased fluorescence intensity and a shift of the spectra to longer emission wavelengths. Decreased fluorescence is due to a drop in detected stromal photons, whereas the shift of spectral shape is attributed to an increased fraction of detected photons arising in the epithelium. PMID- 19123660 TI - Comparison of cerebral oxygen saturation in premature infants by near-infrared spatially resolved spectroscopy: observations on probe-dependent bias. AB - Spatially resolved spectroscopy (SRS) allows the estimation of absolute tissue oxygen saturation, the ratio of oxygenated to total hemoglobin concentration, which may facilitate the comparison of results among patients. Eighty-two premature infants were included over two years. The cerebral tissue oxygenation index (c-TOI) was measured using NIRO 300 (Hamamatsu Photonics KK). c-TOI was measured at several positions in each infant. c-TOI varied over time, increasing in the first third and decreasing in the last third of the study period (p<10( 6)). Two probes were used in the study, and a highly significant difference was found between these (p<10(-6)). The mean difference was 8.5% (95%CI 5.4 to 11.6%). After correction for this difference, there was no variation over time. A conclusive explanation for the bias could not be identified. Since the study groups were well distributed, the bias had no influence on the results of our clinical study. We investigated an unexpected but highly significant probe dependent bias in c-TOI with no conclusive explanation. Hence, comparisons of absolute TOI between groups of patients and among studies should be regarded with caution. A better strategy to detect potential instrumental problems will be useful in preventing biased c-TOI from occurring. PMID- 19123661 TI - Action spectra of electrochromic voltage-sensitive dyes in an intact excitable tissue. AB - Voltage-sensitive dyes (VSDs) provide a spatially resolved optical read-out of electrical signals in excitable tissues. Several common fluorescent VSDs display electrochromic shifts of their emission spectra, making them suitable candidates for ratiometric measurements of transmembrane voltages. These advantages of VSDs are tempered by tissue-specific shifts to their fluorescence emission. In addition, the optimal electrochromic dye response occurs in wavelength bands distinct from the dye's maximal resting emission. This "action spectrum" can undergo tissue-specific shifts as well. We have developed a technique for in situ measurements of the action spectra of VSDs in intact excitable tissues. Fluorescence emission spectra of VSDs during action-potential depolarization were obtained within a single sweep of a spectrophotometer equipped with a change coupled device (CCD) array detector. To resolve the subtle electrochromic shifts in voltage-induced dye emission, fluorescence emission spectra measured right before and during field-induced action-potential depolarization were averaged over about 100 trials. Removing white-noise contributions from the spectrometer's CCD detector/amplifier via low-pass filtering in Fourier space, the action spectra of all dyes could be readily determined. PMID- 19123662 TI - Model-based analysis of reflectance and fluorescence spectra for in vivo detection of cervical dysplasia and cancer. AB - Development, validation, and implementation of an analytical model to extract biologically and diagnostically relevant parameters from measured cervical tissue reflectance and fluorescence spectra are presented. Monte Carlo simulations of tissue reflectance are used to determine the relative contribution of the signal from the epithelium and stroma. The results indicate that the clinical probe used collects a majority of its reflectance signal from the stroma; therefore, a one layer analytical model of reflectance is used. Two analytical approaches to calculate reflectance spectra are compared to Monte Carlo simulations, and a diffusion theory-based model is implemented. The model is validated by fitting spectra generated from Monte Carlo simulations and comparing the input and output parameters. Median agreement between extracted optical properties and input parameters is 10.6%. The reflectance model is used together with an analytical model of tissue fluorescence to extract optical properties and fluorophore concentrations from 748 clinical measurements of cervical tissue. A diagnostic algorithm based on these extracted parameters is developed and evaluated using cross-validation. The sensitivity/specificity of this algorithm relative to the gold standard of histopathology per measurement are 8551%; this is comparable to accuracy reported in other studies of optical technologies for detection of cervical cancer and its precursors. PMID- 19123663 TI - Quantitative and morphometric evaluation of the angiogenic effects of leptin. AB - Angiogenesis is a dynamic process that requires an interaction of pro-and antiangiogenic factors. It is known that the cytokine leptin stimulates endothelial cell growth and angiogenesis, but further quantitative analysis is necessary to understand leptin angiogenic effects. The quail chorioallantoic membrane (CAM) assay has been used to study angiogenesis in vivo by focusing on morphometric parameters that quantify vascular complexity and density. We quantify the angiogenic activity of leptin using the CAM assay by digital morphometry and a computer-assisted image analysis to evaluate more precisely vessel length, diameter, branching, and tortuousity. CAM images are obtained from ex ovo cultures of E8-E9 quail embryos. MATLAB and custom software are used for our analysis. The effects of leptin, vascular endothelial growth factor-165 (VEGF(165)), and their corresponding neutralizing antibodies are compared. Our results show that CAM treated with leptin and VEGF(165) has a significant increase in vascular complexity and density. A corresponding decrease is observed using neutralizing antibodies. Notably, leptin induced more significant changes than VEGF in vessel length and tortuousity. Conversely, VEGF induced a greater increase in vessel branching than leptin. These results underscore the importance of using multiparametric quantitative methods to assess several aspects of angiogenesis and enable us to understand the proangiogenic effects of leptin. PMID- 19123664 TI - Second-harmonic generation and two-photon-excited autofluorescence microscopy of cardiomyocytes: quantification of cell volume and myosin filaments. AB - The ability to quantify changes in cardiomyocyte and myosin volume across gestation and in response to intrauterine insults will lead to a better understanding of the link between low birth weight and an increased risk of heart disease in adult life. We present the use of second-harmonic generation (SHG) and two-photon excitation autofluorescence (TPEF) microscopy to image unstained isolated fetal cardiomyocytes. The simultaneous collection of these two images provides a wealth of information on the morphology of cardiomyocytes. The SHG signal provides high-contrast images of myosin filaments and the TPEF signal can be used to clearly visualize cell morphology. A potential issue may arise if SHG microscopy is performed exclusively due to the lack of sensitivity to distinguish between mononucleated and binucleated cardiomyocytes. However, TPEF microscopy has the ability to efficiently separate the two types of cardiomyocytes. In addition, quantitative analysis of the SHG and TPEF images enables quantification of myosin filament level and accurate determination of cell volume. In short, we demonstrate that advanced nonlinear optical microscopy can be used to answer key physiological questions in the early origins of adult health with increased accuracy and speed compared to previously used methods. PMID- 19123665 TI - Crosstalk and error analysis of fat layer on continuous wave near-infrared spectroscopy measurements. AB - Accurate estimation of concentration changes in muscles by continuous wave near IR spectroscopy for muscle measurements suffers from underestimation and crosstalk problems due to the presence of superficial skin and fat layers. Underestimation error is basically caused by a homogeneous medium assumption in the calculations leading to the partial volume effect. The homogeneous medium assumption and wavelength dependence of mean partial path length in the muscle layer cause the crosstalk. We investigate underestimation errors and crosstalk by Monte Carlo simulations with a three layered (skin-fat-muscle) tissue model for a two-wavelength system where the choice of first wavelength is in the 675- to 775 nm range and the second wavelength is in the 825- to 900-nm range. Means of absolute underestimation errors and crosstalk over the considered wavelength pairs are found to be higher for greater fat thicknesses. Estimation errors of concentration changes for Hb and HbO(2) are calculated to be close for an ischemia type protocol where both Hb and HbO(2) are assumed to have equal magnitude but opposite concentration changes. The minimum estimation errors are found for the 700825- and 725825-nm pairs for this protocol. PMID- 19123666 TI - Photoacoustic monitoring of burn healing process in rats. AB - We performed multiwavelength photoacoustic (PA) measurement for extensive deep dermal burns in rats to monitor the healing process of the wounds. The PA signal peak at 532 nm, an isosbestic point for oxyhemoglobin (HbO(2)) and deoxyhemoglobin (HHb), was found to shift to a shallower region of the injured skin tissue with the elapse of time. The results of histological analysis showed that the shift of the PA signal reflected angiogenesis in the wounds. Until 24 h postburn, PA signal amplitude generally increased at all wavelengths. We speculate that this increase in amplitude is associated with dilation of blood vessels within healthy tissue under the injured tissue layer and increased hematocrit value due to development of edema. From 24 to 48 h postburn, the PA signal showed wavelength-dependent behaviors; signal amplitudes at 532, 556, and 576 nm continued to increase, while amplitude at 600 nm, an HHb absorption dominant wavelength, decreased. This seems to reflect change from shock phase to hyperdynamic state in the rat; in the hyperdynamic state, cardiac output and oxygen consumption increased considerably. These findings show that multiwavelength PA measurement would be useful for monitoring recovery of perfusion and change in local hemodynamics in the healing process of burns. PMID- 19123667 TI - Measuring erythrocyte deformability with fluorescence, fluid forces, and optical trapping. AB - A laser-based method has been developed for experimentally probing single red blood cell (RBC) buckling and determining RBC membrane rigidity. Our method combines a liquid flow cell, fluorescence microscopy, and an optical-trap to facilitate simple measurements of the shear modulus and buckling properties of single RBCs, under physiological conditions. The efficacy of the method is illustrated by studying buckling behavior of normal and Plasmodium-infected RBCs, and the effect of Plasmodium falciparum-conditioned medium on normal, uninfected cells. Our simple method, which quantifies single-RBC deformability, may ease detection of RBC hematological disorders. PMID- 19123668 TI - Intrinsic fluorescence spectroscopy for endoscopic detection and localization of the endobronchial cancerous lesions. AB - Fluorescence spectroscopy contains diagnostic information about the lung biochemistry and morphology, including tissue optical properties and fluorophores. However, the fluorophore information is generally masked by the optical properties of the tissue, which complicates the evaluation of their role in lung-cancer detection. In this work, we have developed a method for extracting the intrinsic fluorescence spectra from the endoscopic measurements of the combined fluorescence and reflectance spectra. Principle components and classification analysis was performed to evaluate the diagnostic potential of the extracted intrinsic fluorescence spectra from in vivo combined fluorescence and reflectance spectral measurements. We evaluated the diagnostic sensitivity and specificity of both the intrinsic fluorescence and the fluorescence spectra. The results showed that the intrinsic fluorescence spectra contain significant diagnostic information that had been masked by the lung optical properties. We have also found that the intrinsic fluorescence has improved the specificity for endobronchial-cancer detection, although with a slight decrease in the detection sensitivity, when compared to the fluorescence spectra. This may indicate that intrinsic fluorescence analysis could be used to improve the diagnostic specificity of fluorescence spectroscopy and imaging. PMID- 19123669 TI - Acute hyperglycemia compromises cerebral blood flow following cortical spreading depression in rats monitored by laser speckle imaging. AB - Hyperglycemia and cortical spreading depression (CSD) are possible factors that worsen the outcome of ischemic stroke, and it is probable that there is a longterm cooperative effect of hyperglycemia and CSD on cerebral blood flow (CBF). Long-lasting and full-field observation of changes in CBF following CSD in vivo during acute hyperglycemia in rats might show whether this is the case. Here, we utilized laser speckle imaging to study influences of acute hyperglycemia on CBF at the level of individual vascular compartments for 3 h in normal rats and those with CSD. It is shown that there are extensive increases of CBF at the arteriole and parenchyma over the normal rat cortex during acute hyperglycemia, whereas there is no significant change in CBF at the venule. We also find that, at all vascular compartments, after the glucose administration there is a stepwise reduction of CBF following CSD, but after saline injection CBF following CSD is close to the baseline. Our results indicate that acute hyperglycemia could aggravate the severity of decrease in CBF following CSD, suggesting possible mechanisms by which hyperglycemia exacerbates cerebral damage after ischemic stroke. PMID- 19123670 TI - New laryngoscope for quantitative high-speed imaging of human vocal folds vibration in the horizontal and vertical direction. AB - We report the design of a novel laser line-triangulation laryngoscope for the quantitative visualization of the three-dimensional movements of human vocal folds during phonation. This is the first successful in vivo recording of the three-dimensional movements of human vocal folds in absolute values. Triangulation images of the vocal folds are recorded at the rate of 4000 fps with a resolution of 256x256 pixels. A special image-processing algorithm is developed to precisely follow the subpixel movements of the laser line image. Vibration profiles in both horizontal and vertical directions are calibrated and measured in absolute SI units with a resolution of +/-50 microm. We also present a movie showing the vocal folds dynamics in vertical cross section. PMID- 19123671 TI - Detection of optical and mechanical property inhomogeneities in tissue mimicking phantoms using an ultrasound assisted optical probe. AB - We discuss the issue of separating contributions from mechanical and optical properties of a moderately scattering tissue phantom to the modulation depth (M) of intensity autocorrelation measured in an ultrasound-assisted optical tomography system using axial and transverse illuminations. For axial illumination, M is affected by both the displacement and absorption coefficient, more prominently by displacement. But transverse illumination has very little contribution from displacement of scattering centers. Since displacement is related to the elastic property of the insonified region, we show that there is a possibility of separating the contributions from elastic and optical properties of the insonified region using axial and transverse illuminations. The main conclusions of our study using moderately scattering phantoms are: 1. axial illumination is the best for mapping storage modulus inhomogeneities, but M is also affected by optical absorption; 2. transverse illumination is the best for mapping absorption inhomogeneities; and 3. for the practically relevant case of an inclusion with larger storage modulus and absorption, both illuminations produced large contrast in M. When the scattering coefficient is high, the angle dependence of illumination is lost and the present method is shown to fail to separate these contributions based on direction of illumination. PMID- 19123672 TI - Quantitative depth analysis of optic nerve head using stereo retinal fundus image pair. AB - Depth analysis of the optic nerve head (ONH) in the retinal fundus is important for the early detection of glaucoma. In this study, we investigate an automatic reconstruction method for the quantitative depth measurement of the ONH from a stereo retinal fundus image pair. We propose a technique to obtain the depth value from the stereo retinal fundus image pair, which mainly consists of five steps: 1. cutout of the ONH region from the stereo retinal fundus image pair, 2. registration of the stereo image pair, 3. disparity measurement, 4. noise reduction, and 5. quantitative depth calculation. Depth measurements of 12 normal eyes are performed using the stereo fundus camera and the Heidelberg Retina Tomograph (HRT), which is a confocal laser-scanning microscope. The depth values of the ONH obtained from the stereo retinal fundus image pair were in good accordance with the value obtained using HRT (r=0.80+/-0.15). These results indicate that our proposed method could be a useful and easy-to-handle tool for assessing the cup depth of the ONH in routine diagnosis as well as in glaucoma screening. PMID- 19123674 TI - In vivo staining of neocortical astrocytes via the cerebral microcirculation using sulforhodamine B. AB - Staining and imaging glial cells in vivo while observing the microvasculature could help understand brain physiology, namely neuronal-glial-vascular communication. Two-photon excitation microscopy provides a means to monitor these interactions at the cellular level in living animals, but the cells of interest must be fluorescent. Injecting dyes intravenously is a rapid and quasi noninvasive method to stain cells in the brain. It necessitates that the dye is soluble in the blood plasma and crosses the blood brain barrier (BBB). We demonstrate here, using two-photon imaging, that sulforhodamine B (SRB) crosses the BBB and stains in vivo, specifically mouse astrocytes. This is confirmed by experiments on primary neurons and astrocytes cultures showing the preferential SRB staining of the latter. SRB is rapidly eliminated from the blood, which allows repeated injections in longitudinal studies. PMID- 19123673 TI - Tissue hemoglobin monitoring of progressive central hypovolemia in humans using broadband diffuse optical spectroscopy. AB - We demonstrate noninvasive near-infrared diffuse optical spectroscopy (DOS) measurements of tissue hemoglobin contents that can track progressive reductions in central blood volume in human volunteers. Measurements of mean arterial blood pressure (MAP), heart rate (HR), stroke volume (SV), and cardiac output (Q) are obtained in ten healthy human subjects during baseline supine rest and exposure to progressive reductions of central blood volume produced by application of lower body negative pressure (LBNP). Simultaneous quantitative noninvasive measurements of tissue oxyhemoglobin (OHb), deoxyhemoglobin (RHb), total hemoglobin concentration (THb), and tissue hemoglobin oxygen saturation (S(t)O(2)) are performed throughout LBNP application using broadband DOS. As progressively increasing amounts of LBNP are applied, HR increases, and MAP, SV, and Q decrease (p<0.001). OHb, S(t)O(2), and THb decrease (p<0.001) in correlation with progressive increases in LBNP, while tissue RHb remained relatively constant (p=0.378). The average fractional changes from baseline values in DOS OHb (fOHb) correlate closely with independently measured changes in SV (r(2)=0.95) and Q (r(2)=0.98) during LBNP. Quantitative noninvasive broadband DOS measurements of tissue hemoglobin parameters of peripheral perfusion are capable of detecting progressive reductions in central blood volume, and appear to be sensitive markers of early hypoperfusion associated with hemorrhage as simulated by LBNP. PMID- 19123675 TI - Image reconstruction method for a two-layer tissue structure accounts for chest wall effects in breast imaging. AB - We develop a new tomographic imaging reconstruction algorithm for a two-layer tissue structure. Simulations and phantom experiments show more accurate reconstruction of target optical properties compared with those results obtained from a semi-infinite tissue model for layered structures. This improvement is mainly attributed to the more accurate estimation of background optical properties and more accurate estimation of weight matrix for imaging reconstruction by considering the light propagation effect in the second layer. Clinical results of breast lesions are also presented to demonstrate the utility of this new imaging algorithm. PMID- 19123676 TI - Estimation of mammographic density on an interval scale by transillumination breast spectroscopy. AB - Transillumination breast spectroscopy (TiBS) uses nonionizing optical radiation to gain information about breast tissue morphological and structural properties. TiBS spectra are obtained from 232 women and compared to mammographic density (MD) quantified using Cumulus. The ability of TiBS to estimate MD is assessed using partial least-squares (PLS) regression methods, which requires TiBS spectra as input (X) and Cumulus MD as target (Y) data. Multiple PLS models are considered to determine the optimal processing technique(s) for the input (X) and target (Y) data. For each model, the association between TiBS estimated MD (Y) and Cumulus MD (Y) is established using Spearman's rank correlation and linear regression analysis. The model that best estimates MD has the fewest assumptions regarding target (Y) and spectral (X) processing. The Spearman's correlation coefficient between predicted MD and Cumulus MD for this model is 0.88, with a regression slope (beta) of 0.93 (95% CI 0.83-1.02) and an R(2) of 0.78. The approximation of individual MD was within 10% of Cumulus MD for the majority of women (80%), without stratification on age, body mass index (BMI), and menopausal status. TiBS provides an alternative to mammography assessed MD enabling frequent and earlier use of MD as a risk marker in preventive oncology. PMID- 19123677 TI - Imaging of zinc oxide nanoparticle penetration in human skin in vitro and in vivo. AB - Zinc oxide (ZnO-nano) and titanium dioxide nanoparticles (20 to 30 nm) are widely used in several topical skin care products, such as sunscreens. However, relatively few studies have addressed the subdermal absorption of these nanoparticles in vivo. We report on investigation of the distribution of topically applied ZnO in excised and in vivo human skin, using multiphoton microscopy (MPM) imaging with a combination of scanning electron microscopy (SEM) and an energy-dispersive x-ray (EDX) technique to determine the level of penetration of nanoparticles into the sub-dermal layers of the skin. The good visualization of ZnO in skin achieved appeared to result from two factors. First, the ZnO principal photoluminescence at 385 nm is in the "quiet" spectral band of skin autofluorescence dominated by the endogenous skin fluorophores, i.e., NAD[P]H and FAD. Second, the two-photon action cross section of ZnO-nano [sigma(ZnO) ((TPEF)) approximately 0.26 GM; diameter, 18 nm] is high: approximately 500-fold of that inferred from its bulk third-order nonlinear susceptibility [Im chi(ZnO) ((3))], and is favorably compared to that of NAD[P]H and FAD. The overall outcome from MPM, SEM, and EDX studies was that, in humans in vivo, ZnO nanoparticles stayed in the stratum corneum (SC) and accumulated into skin folds and/or hair follicle roots of human skin. Given the lack of penetration of these nanoparticles past the SC and that the outermost layers of SC have a good turnover rate, these data suggest that the form of ZnO-nano studied here is unlikely to result in safety concerns. PMID- 19123678 TI - Quantitative real-time analysis of nucleolar stress by coherent phase microscopy. AB - We develop a method of coherent phase microscopy (CPM) for direct visualization of nonfixed, nonstained mammalian cells (both cultured cells and freshly isolated tumor biopsies) followed by computer-assisted data analysis. The major purpose of CPM is to evaluate the refractive properties of optically dense intracellular structures such as the nucleus and the nucleoli. In particular, we focus on quantitative real-time analysis of the nucleolar dynamics using phase thickness as an equivalent of optical path difference for optically nonhomogenous biological objects. Pharmacological inhibition of gene transcription leads to a dramatic decrease of the phase thickness of the nucleoli within the initial minutes of cell exposure. Furthermore, the acute depletion of intracellular ATP pool, depolymerization of microtubules and inhibition of DNA replication resulted in a rapid decrease of the nucleolar phase thickness. These optical effects were paralleled by segregation of nucleolar components as documented by electron microscopy. Thus, CPM detects early changes of nucleolar dynamics, in particular, the nucleolar segregation as part of general cellular response to cytotoxic stress, regardless of whether the nucleolus is or is not the primary target of the toxin. CPM is applicable for monitoring and quantitative analysis of the "nucleolar stress" in living mammalian cells. PMID- 19123679 TI - Thin polymer etalon arrays for high-resolution photoacoustic imaging. AB - Thin polymer etalons are demonstrated as high-frequency ultrasound sensors for three-dimensional (3-D) high-resolution photoacoustic imaging. The etalon, a Fabry-Perot optical resonator, consists of a thin polymer slab sandwiched between two gold layers. It is probed with a scanning continuous-wave (CW) laser for ultrasound array detection. Detection bandwidth of a 20-microm-diam array element exceeds 50 MHz, and the ultrasound sensitivity is comparable to polyvinylidene fluoride (PVDF) equivalents of similar size. In a typical photoacoustic imaging setup, a pulsed laser beam illuminates the imaging target, where optical energy is absorbed and acoustic waves are generated through the thermoelastic effect. An ultrasound detection array is formed by scanning the probing laser beam on the etalon surface in either a 1-D or a 2-D configuration, which produces 2-D or 3-D images, respectively. Axial and lateral resolutions have been demonstrated to be better than 20 microm. Detailed characterizations of the optical and acoustical properties of the etalon, as well as photoacoustic imaging results, suggest that thin polymer etalon arrays can be used as ultrasound detectors for 3-D high resolution photoacoustic imaging applications. PMID- 19123680 TI - Model-based spectroscopic analysis of the oral cavity: impact of anatomy. AB - In order to evaluate the impact of anatomy on the spectral properties of oral tissue, we used reflectance and fluorescence spectroscopy to characterize nine different anatomic sites. All spectra were collected in vivo from healthy oral mucosa. We analyzed 710 spectra collected from the oral cavity of 79 healthy volunteers. From the spectra, we extracted spectral parameters related to the morphological and biochemical properties of the tissue. The parameter distributions for the nine sites were compared, and we also related the parameters to the physical properties of the tissue site. k-Means cluster analysis was performed to identify sites or groups of sites that showed similar or distinct spectral properties. For the majority of the spectral parameters, certain sites or groups of sites exhibited distinct parameter distributions. Sites that are normally keratinized, most notably the hard palate and gingiva, were distinct from nonkeratinized sites for a number of parameters and frequently clustered together. The considerable degree of spectral contrast (differences in the spectral properties) between anatomic sites was also demonstrated by successfully discriminating between several pairs of sites using only two spectral parameters. We tested whether the 95% confidence interval for the distribution for each parameter, extracted from a subset of the tissue data could correctly characterize a second set of validation data. Excellent classification accuracy was demonstrated. Our results reveal that intrinsic differences in the anatomy of the oral cavity produce significant spectral contrasts between various sites, as reflected in the extracted spectral parameters. This work provides an important foundation for guiding the development of spectroscopic-based diagnostic algorithms for oral cancer. PMID- 19123681 TI - Pulsed diode laser-based singlet oxygen monitor for photodynamic therapy: in vivo studies of tumor-laden rats. AB - Photodynamic therapy (PDT) is a promising cancer treatment that involves optical excitation of photosensitizers that promote oxygen molecules to the metastable O(2)(a(1)Delta) state (singlet oxygen). This species is believed to be responsible for the destruction of cancerous cells during PDT. We describe a fiber optic-coupled, pulsed diode laser-based diagnostic for singlet oxygen. We use both temporal and spectral filtering to enhance the detection of the weak O(2)(a-->X) emission near 1.27 microm. We present data that demonstrate real-time singlet oxygen production in tumor-laden rats with chlorin e6 and 5 aminolevulinic acid-induced protoporphyrin photosensitizers. We also observe a positive correlation between post-PDT treatment regression of the tumors and the relative amount of singlet oxygen measured. These results are promising for the development of the sensor as a real-time dosimeter for PDT. PMID- 19123682 TI - Bioimaging assessment and effect of skin wound healing using bone-marrow-derived mesenchymal stromal cells with the artificial dermis in diabetic rats. AB - We investigate the relationship between the fate and healing effect of transplanted mesenchymal stromal cells (MSCs) in a rat diabetic skin wound model. Rats are treated with streptozotocin to induce diabetic conditions. A full thickness skin defect is surgically made on the head of diabetic rats, and covered with an artificial dermis impregnated with either bone marrow cells (BMCs) or bone-marrow-derived MSCs from firefly luciferase (luc) transgenic (Tg) rats. Wound healing is evaluated using planimetry and immunohistochemistry, and the fate of transplanted MSCs is determined using in-vivo luminescent imaging. The diabetic wound treated with MSCs-impregnated artificial dermis is significantly smaller than that treated with artificial dermis alone at 1 week postoperation. Photons of luc+ MSCs are detected at the transplanted site during healing (3 weeks), whereas those of luc+ MSCs are depleted only after 1 week postimplantation. Immunohistochemistry at the healing site treated with MSCs demonstrates that CD31+ vessels increase with expression of vascular endothelial growth factor, suggesting that MSCs accelerate angiogenesis. These findings suggest that transplanted MSCs could be retained at wound sites during the healing process in a diabetic rat model, and subsequently promote wound healing through angiogenesis. PMID- 19123683 TI - Diffusion measured by fluorescence recovery after photobleaching based on multiphoton excitation laser scanning microscopy. AB - Fluorescence recovery after photobleaching (FRAP) is a widely used method to measure diffusion. The technique is normally based on one-photon excitation, which limits diffusion to two dimensions due to extended photobleaching in the axial direction. Multiphoton excitation, on the other hand, creates a well defined focal volume. In the present work, FRAP based on a scanning laser beam and two-photon excitation is used to measure diffusion of macromolecules in solution and gels, as well as in the extracellular matrix in multicellular spheroids and tumor tissue in dorsal chambers. The bleaching profile is determined experimentally in immobilized gels, and for small scanning areas (approximately twice the lateral radius of the laser beam) a Gaussian bleaching distribution is found. In addition, the bleaching profile is determined theoretically based on the convolution of the Gaussian point spread function and a circular scanning area. The diffusion coefficient is determined by fitting a mathematical model based on a Gaussian laser beam profile to the experimental recovery curve. The diffusion coefficient decreases with increasing complexity of the sample matrix and increasing the amount of collagen in the gels. The potential of using two-photon laser scanning microscopes for noninvasive diffusion measurements in tissue is demonstrated. PMID- 19123684 TI - Analysis of the fluorescence temporal point-spread function in a turbid medium and its application to optical imaging. AB - A time-domain optical method to evaluate the concentration (n), lifetime (tau), and depth (d) of a fluorescent inclusion is described by the complete analysis of the fluorescence temporal point-spread function (TPSF). The behavior of parameters in the fluorescence TPSF is explored, and we demonstrate the method with experimental data from a localized fluorescent inclusion in scattering media to recover images of n, tau, and d. The method has potential application for in vivo fluorescence imaging. PMID- 19123686 TI - Improving surface-enhanced Raman scattering effect using gold-coated hierarchical polystyrene bead substrates modified with postgrowth microwave treatment. AB - We report a novel postgrowth microwave heating implementation by selectively modifying hierarchical polystyrene (PS) bead substrates coated with gold (Au) films to effectively improve the surface-enhanced Raman scattering (SERS) effect on the analytes. The SERS signal of probe molecule rhodamine 6G (Rh 6G) on the microwave-treated Au-PS substrates can be improved by 10-fold, while the detection limit of Rh 6G in concentration can be enhanced by two orders of magnitude compared to the as-growth substrates. The high-quality SERS spectrum of saliva can also be acquired using the modified substrates, demonstrating the potential for the realization of the high-performance SERS substrates for biomedical applications. PMID- 19123685 TI - Monitoring tau-tubulin interactions utilizing second harmonic generation in living neurons. AB - Tau is a microtubule associated protein that is localized to the axon in neurons. During pathological conditions, including frontotemporal dementia (FTD), a shift in tau isoforms occurs that leads to enhanced expression of a form of tau with four (rather than three) microtubule binding repeats; this has been postulated to alter microtubule structure. Second harmonic generation (SHG) is a technique that allows the visualization of intact microtubules in axons of living neurons without the need for labeling or fixing. We examined how the presence of exogenous tau influences SHG in living neurons. Our results show that the presence of tau significantly enhances SHG, specifically in neuronal axons, despite the presence of tau throughout the entire cell. Our data also suggest that the presence or absence of the fourth microtubule binding repeat does not significantly alter tau's ability to enhance SHG. These results provide evidence that SHG is a useful, noninvasive tool to study tau-microtubule interactions in axons; further, it appears that tau overexpression, rather than specific isoforms, is the major contributor to tau-induced changes in axonal microtubule SHG signal. PMID- 19123687 TI - In vivo long-term continuous observation of gene expression in zebrafish embryo nerve systems by using harmonic generation microscopy and morphant technology. AB - Gene expression plays an important role in embryo development and organ function. Previous studies have shown that harmonic generation microscopy (HGM) can be used as a fluorescence signal-independent, minimally invasive method with a subcellular 3-D resolution and a penetration depth in the order of millimeters for long-term continuous imaging of vertebrate embryos. We show that it is ideal to combine in vivo HGM with the morphant technology for minimally invasive, long term continuous observation of gene expression in the nervous system of vertebrate embryos. Since second- and third-harmonic generations (SHG, THG) are virtual-state-transition-based systems that depend only on the structure of the organisms, they are not temporally limited by the expression of the fluorescence proteins. We successfully identified the expression of the zarnt2a and the hif 1alpha, 2alpha, and 3alpha genes in the nervous system of zebrafish embryos with specific knockdown genes by microscopically observing the embryos from the early stages of embryogenesis. The results from a combination of the two different modalities, i.e., SHG microscopy and THG microscopy, successfully revealed the weak cell adhesion, cell apoptosis, nerve formation reduction, and neural tube distortion in the morphant zebrafish embryos. PMID- 19123688 TI - Empirical model description of photon path length for differential path length spectroscopy: combined effect of scattering and absorption. AB - Differential path length spectroscopy (DPS) is a method of reflectance spectroscopy that utilizes a specialized fiber geometry to make the photon path length (tau) insensitive to variations in tissue optical properties over a wide range of absorption (mu(a)) and total scattering (mu(s)) coefficients, which are common within the ultraviolet/visible (UV/VIS) wavelength region. This study extends the description of tau to larger mu(a) and smaller mu(s) values, optical properties that are representative of the near-infrared region (NIR), a region where the DPS path length may be dependent on both coefficients. This study presents a novel empirical relationship between tau and the combined effect of both mu(a) (range: 0.1-12 mm(-1)) and mu(s) (range: 1.5-42 mm(-1)), anisotropy of 0.8, and is applicable to DPS probes containing a wide range of fiber diameters (range: 100-1000 microm). The results indicate that the simple empirical formula, including only one fitted parameter, is capable of accurately predicting tau over a wide range (r=0.985; range: 80-940 microm) and predictions are not biased versus mu(a) or mu(s). This novel relationship is applicable to analysis of DPS measurements of tissue in both the UV/VIS and NIR wavelength regions and may provide information about the wavelength-specific tissue volume optically sampled during measurement. PMID- 19123689 TI - Monitoring angiogenesis noninvasively with near-infrared spectroscopy. AB - Near-infrared (NIR) spectroscopy is used to quantify cerebral blood volume (CBV) as a marker of angiogenesis (formation of new blood vessels). Rats are exposed to chronic hypoxia for 3 weeks at half atmospheric pressure to stimulate angiogenesis, and second-differential NIR spectroscopy is used to quantify total cerebral hemoglobin before and after angiogenesis. The cerebral hemoglobin (from broadband NIR spectroscopy), and the large vessel hemoglobin and hematocrit (from blood samples), are used to derive values for the calculation of CBV. The total hemoglobin in brain is 46.6+/-1.9 micromoll (mean+/-SD, n=5) preacclimation and increases by 72% postacclimation. CBV is initially 3.26+/-0.41% v/v and increases by 31% with acclimation. Each individual animal shows a measureable increase in CBV. This study indicates that NIR broadband spectroscopy can be used for repeated measurements of CBV and can be applied as a noninvasive method to study angiogenesis. PMID- 19123692 TI - Pulp vitality in teeth suffering trauma during orthodontic therapy. AB - OBJECTIVE: To examine pulpal vitality in teeth suffering dental trauma during orthodontic therapy with fixed appliances. MATERIALS AND METHODS: Pulpal condition was evaluated in 59 teeth that had suffered dental trauma during orthodontic treatment (TO-group), in 800 orthodontically treated teeth without previous dental trauma (O-group), and in 193 orthodontically untreated teeth with previous dental trauma (T-group). Pulpal vitality was examined clinically and with radiographs. Degree of pulp obliteration was rated as absent, partial, or total. All teeth in the TO-group showed a positive sensibility test prior to resumption of orthodontic therapy. RESULTS: Teeth in the TO-group revealed a significantly higher frequency of pulp necrosis than teeth in the O-group or teeth in the T-group (P < .001, respectively). In the TO-group, teeth with extrusive or lateral luxation (P = .031) and teeth with intrusive luxation (P = .015) injuries showed a significantly higher rate of pulp necrosis than teeth with fracture of enamel. In addition, teeth with total pulp obliteration showed a significantly higher frequency of pulp necrosis than teeth without pulp obliteration (P = .013). CONCLUSION: Teeth with severe periodontal injury during orthodontic therapy and subsequent total pulp obliteration have an increased risk of pulp necrosis during additional orthodontic treatment stages. PMID- 19123693 TI - Success rate of second-generation palatal implants. AB - OBJECTIVE: To analyze the clinical outcome of a prospective two-center study of second-generation palatal implants 6 months after functional loading. MATERIAL AND METHODS: From 2005 to 2006, 30 patients aged 12 to 41 years were included in the study. In all patients, orthodontic treatment required stationary anchorage. The palatal implants (Straumann, Basel, Switzerland) were placed in the median region of the anterior palate. RESULTS: All implants were initially stable at the time of placement. However, two (6.7%) were lost during the unloaded healing period. The remaining 28 (93.3%) were subjected to functional loading after a mean healing period of 12 weeks. Typical signs of slight superficial inflammation were observed in the peri-implant mucosa (n = 28). During the orthodontic loading phase, the implants were equipped with either a modified pendulum appliance for distalization or a transpalatal arch for stationary anchorage to the posterior teeth. No implant loosening or loss was registered during the active treatment period. CONCLUSIONS: The failure rate of palatal implants of the second generation was low (6.7%). Slight inflammatory reactions of peri-implant tissue caused neither implant loss nor pain. PMID- 19123694 TI - Why do dentists refer to specific orthodontists? AB - OBJECTIVE: This study tested the hypothesis that the quality of treatment outcomes and overall patient satisfaction are of equal importance in influencing a general dentist to refer a patient to an orthodontist for treatment. The investigation also attempted to determine what specific aspects of an orthodontically treated occlusion comprise an outstanding result in the opinion of the general dentist. MATERIALS AND METHODS: Self-administered surveys containing 35 questions were distributed to a random sample of 1000 general dentists in the midwestern United States. RESULTS: Three-quarters responded that the quality of previous orthodontic treatment and patient/ parental satisfaction were of equal importance in the orthodontic referral decision. If additional factors were considered, the quality of the treated result as judged by the general dentist was far more important than the location of the orthodontist's office, the reputation of the orthodontist, or the anticipated cost of treatment. With regard to desired postorthodontic occlusal and functional characteristics, the majority of the dentists ranked canine guidance as most important. Other characteristics were Class I molar and canine relationships, even contact of all teeth in centric occlusion, amount of overjet and overbite, absence of spacing, and absence of balancing interferences. CONCLUSIONS: The decision of the general dentist to refer a patient to one orthodontist over another is based on a number of interactive factors. It behooves the orthodontic clinician to provide high quality treatment, to interact well with his or her patients and their families, and to maintain good communications with general practitioners in the community. PMID- 19123695 TI - Immediate and post-retention effects of rapid maxillary expansion investigated by computed tomography in growing patients. AB - OBJECTIVE: To determine by low-dose computed tomography (CT) protocol the dental and periodontal effects of rapid maxillary expansion (RME). MATERIALS AND METHODS: The sample comprised 17 subjects (7 males and 10 females), with a mean age at first observation of 11.2 years. Each patient underwent expansion of 7 mm. Multislice CT scans were taken before rapid palatal expansion (T0), at the end of the active expansion phase (T1), and after a retention period of 6 months (T2). On scanned images, measurements were performed at the dental and periodontal levels. Mean differences between measurements at T0, T1, and T2 were examined through analysis of variance (ANOVA) for repeated measures with post-hoc tests. RESULTS: All interdental transverse measurements were significantly increased at both T1 and T2 with respect to T0. In the evaluation of T0-T1 changes, periodontal measurements were significant on the buccal aspect of banded teeth with a reduction in alveolar bone thickness corresponding to the mesial (-0.5 mm; P < .05) and distal (-0.4 mm; P < .05) roots of the right first molar and to the mesial root of the left first molar (-0.3 mm; P < .05). In the evaluation of overall T0-T2 changes, the lingual bone plate thickness of both first molars was found to be significantly increased (+0.6 mm; P < .05). CONCLUSIONS: RME therapy induces a significant increase in the transverse dimension of the maxillary arch in growing subjects without causing permanent injury to the periodontal bony support of anchoring teeth discernible on CT imaging. PMID- 19123696 TI - Water and saliva contamination effect on shear bond strength of brackets bonded with a moisture-tolerant light cure system. AB - OBJECTIVE: To evaluate the effects of water and saliva contamination on shear bond strength of brackets bonded with a moisture-tolerant light cure system. MATERIALS AND METHODS: Brackets were bonded to 240 bovine lower incisors divided into 12 groups. Four bonding procedures were evaluated, including (1) TSEP/Transbond XT, (2) TMIP/ Transbond XT, (3) TSEP/Transbond PLUS, and (4) TMIP/Transbond PLUS, each under three different bonding conditions: without contamination, with water contamination, and with saliva contamination. Shear bond strength was measured with a universal testing machine. The adhesive remnant on the teeth was quantified with the use of image analyzing equipment. RESULTS: Without contamination, bond strengths for the four procedures were similar (P > .05). TSEP/Tranbond PLUS and TMIP/Transbond PLUS left significantly less adhesive on the teeth after debonding than TSEP/Transbond XT and TMIP/Transbond XT (P < .008). Bond strength and adhesive remaining for TMIP/Transbond XT contaminated with water or saliva showed significantly worse performance than the other procedures evaluated (P < .008). Contamination (with water or saliva) did not affect either bond strength or adhesive remaining on the teeth for TSEP/ Transbond XT, TSEP/Transbond PLUS, or TMIP/Transbond PLUS (P > .017), although for TMIP/ Transbond XT, both variables showed significant reductions after contamination (P < .017). CONCLUSION: TSEP/Transbond PLUS, TMIP/Transbond PLUS, and TSEP/Transbond XT showed greater tolerance to wet conditions than was shown by TMIP/Transbond XT. PMID- 19123697 TI - Is Herbst-multibracket appliance treatment more efficient in adolescents than in adults? AB - OBJECTIVE: To determine whether Herbst treatment is more efficient in adolescent than in adult Class II division 1 subjects. MATERIALS AND METHODS: All Class II division 1 patients with a full secondary dentition who had been treated at the orthodontic department of the University of Giessen with a Herbst appliance between 1990 and 2000 were considered. The complete records of 77 patients were available. According to their skeletal maturity, as assessed on hand-wrist radiographs, the subjects were divided into an adolescent group (MP3-F to MP3-H; n = 49; mean age 13.5 years) and an adult group (R-IJ to R-J; n = 28; mean age 20.7 years). Pretreatment and posttreatment dental casts were evaluated using the Peer Assessment Rating (PAR) Index. The reductions in PAR scores of the two groups were compared. RESULTS: Before treatment, both groups had a severe Class II division 1 malocclusion. The average PAR score of the adolescent patients was slightly lower (27.8) than that of the adult patients (28.8). After treatment, good results were reached for both groups, and the average PAR scores of the two groups were comparable (adolescents: 4.5; adults: 4.8). The average reductions in PAR score were 82.7% (23.3 points) for the adolescent group and 82.9% (24.0 points) for the adults, indicating great improvement in both groups. CONCLUSIONS: Because good treatment results were achieved, with substantial improvement of the pretreatment situation in both groups, Herbst treatment can be considered equally efficient in adolescent and in adult Class II division 1 subjects. PMID- 19123698 TI - Distal movement of maxillary molars using miniscrew anchorage in the buccal interradicular region. AB - OBJECTIVE: To quantify the treatment effects of interradicular miniscrew anchorage and to confirm the validity of the clinical usage of interradicular miniscrews in the distal movement of maxillary molars in nonextraction treatment. MATERIALS AND METHODS: Twenty-four maxillary molars were moved to the distal using miniscrews placed in the interradicular space between the second premolar and the first molar at an oblique angle of 20 to 30 degrees to the long axis of the proximal tooth. The teeth were evaluated as to how the molars were moved to the distal with the use of lateral cephalograms and dental casts. RESULTS: Maxillary molars were moved to the distal by 2.8 mm with distal tipping of 4.8 degrees and intruded by 0.6 mm. Maxillary incisors were moved to the distal by 2.7 mm with palatal tipping of 4.3 degrees. Molar extrusion and/or consequent mandibular rotation was not observed in any patient. CONCLUSION: Miniscrews placed in the maxillary interradicular space provide successful molar distal movement of 2.8 mm without patient compliance and with no undesirable side effects such as incisor proclination, clockwise mandibular rotation, or root resorption. PMID- 19123699 TI - Structure, composition, and mechanical properties of Australian orthodontic wires. AB - OBJECTIVE: To investigate the surface morphology, structure, elemental composition, and key mechanical properties of various sizes and tempers of Australian wires. MATERIALS AND METHODS: Three types of Australian wire were used: 0.016'' regular, 0.018'' regular+, and 0.018'' special+ (A.J. Wilcock, Whittlesea, Victoria, Australia). Each type of wire was subjected to scanning electron microscopy (SEM) analysis, x-ray energy dispersive spectroscopy (EDS) investigation, Vickers hardness testing, and tensile testing. The modulus of elasticity and ultimate tensile strength were determined. Hardness, modulus, and strength data were analyzed with one-way analysis of variance (ANOVA) and Tukey testing at the .05 level of significance. RESULTS: All three types of Australian wire were found to possess considerably rough surfaces with striations, irregularities, and excessive porosity. All three wire types had high levels of carbon and a similar hardness, which ranged within 600 VHN (Vickers hardness number), and a similar modulus of elasticity (173 to 177 GPa). The 0.018'' special+ had a significantly lower tensile strength (1632 MPa) than the 0.016'' regular and the 0.018'' regular+ wire (2100 MPa). CONCLUSIONS: Australian wires did not show variation implied by the size or temper of the wires. PMID- 19123700 TI - Self-concept and the perception of facial appearance in children and adolescents seeking orthodontic treatment. AB - OBJECTIVE: To examine, in adolescents with mild to moderate malocclusion, the relationship between self-concept and demographic characteristics, a clinical assessment of malocclusion, self-perception of malocclusion, and self-perception of facial attractiveness. METHODS AND MATERIALS: Fifty-nine consecutive patients ages 9 to 15 years scheduled for initial records in a graduate orthodontic clinic consented to participate. Each subject independently completed the Multidimensional Self-Concept Scale (MSCS), the Facial Image Scale, and the Index of Treatment Need-Aesthetic Component (IOTN-AC). Peer Assessment Rating (PAR) scores were obtained from the patients' diagnostic dental casts. Forward multiple regression analysis with a backward overlook was used to analyze the effect of the demographic, clinical, and self-perception measures on each of the six self concept (MSCS) domains. RESULTS: Self-perception of the dentofacial region was the only statistically significant predictor (P < .05) for the Global, Competence, Affect, Academic, and Physical domains of self-concept, while age, parental marital status, and the adolescent's self-perception of the dentofacial region were statistically significant predictors (P < .05) of Social Self Concept. CONCLUSION: The self-perceived level of the attractiveness or "positive" feelings toward the dentofacial region is more strongly related to self-concept than the severity of the malocclusion as indicated by the PAR score or by the adolescent's perception of their malocclusion. PMID- 19123701 TI - Bond strength of metallic brackets after dental bleaching. AB - OBJECTIVE: To test the hypothesis that there is no difference between the action of at-home and in-office vital bleaching on the shear bond strength of metallic brackets bonded with composite resin 24 hours after bleaching. MATERIALS AND METHODS: Forty-five human upper premolars were randomly divided into three groups: (1) control, (2) 10% carbamide peroxide at-home bleached, and (3) 35% hydrogen peroxide in-office bleached. Twenty-four hours after bleaching the teeth were pumiced, bonded with metallic brackets, and stored in distilled water. One day after bonding the shear bond strength of the brackets was determined. RESULTS: The mean shear bond strength of Group 1 (control) and Group 2 (carbamide peroxide bleached) were not statistically different. Group 3 (hydrogen peroxide bleached) had a significantly lower mean shear bond strength than Group 1 and Group 2. CONCLUSIONS: The hypothesis is rejected. Use of 10% carbamide peroxide bleaching does not significantly alter shear bond strength values. On the other hand, use of 35% hydrogen peroxide bleaching significantly reduces these values and diminishes the amount of resin remnant on the tooth surface after bracket debonding. PMID- 19123702 TI - Adhesives for fixed orthodontic bands. A systematic review. AB - OBJECTIVE: To evaluate the effectiveness of adhesives used to attach bands to teeth during fixed appliance treatment. MATERIAL AND METHODS: Electronic databases, conference proceedings and the Internet were searched. There was no restriction with regard to publication status or language of publication. Randomized controlled trials (RCTs) and controlled clinical trials (CCTs) (including split-mouth studies) of adhesives used to attach orthodontic bands to molar teeth were selected. Patients with full arch fixed orthodontic appliance(s) who had bands attached to molars were included. All review authors were involved in study selection, validity assessment, and data extraction. Disagreements were resolved by discussion. Comparisons were made between the main types of adhesive. RESULTS: Five RCTs and three CCTs were identified, all of split-mouth design. Four trials compared chemically cured zinc phosphate and chemically cured glass ionomer; three trials compared chemically cured glass ionomer cement with light cured compomer; and one trial compared chemically cured glass ionomer with a chemically cured glass polyphosphonate. Data analysis was often inappropriate within the studies. Meta-analysis was not feasible. CONCLUSIONS: There is insufficient high-quality evidence with regard to the most effective adhesive for attaching orthodontic bands to molar teeth. Further RCTs are required. PMID- 19123703 TI - Type of archwire and level of acidity: effects on the release of metal ions from orthodontic appliances. AB - OBJECTIVE: To examine the effects of three different parameters-pH value, type of archwire, and length of immersion-on release of metal ions from orthodontic appliances. MATERIALS AND METHODS: Simulated fixed orthodontic appliances that corresponded to one-half of the maxillary arch were immersed in artificial saliva of different pH values (6.75 +/- 0.15 and 3.5 +/- 0.15) during a 28-day period. Three types of archwires were used: stainless steel (SS), nickel-titanium (NiTi), and thermo NiTi. The quantity of metal ions was determined with the use of a high resolution mass spectrophotometer (HR-ICP/MS). RESULTS: The release of six different metal ions was observed: titanium (Ti), chromium (Cr), nickel (Ni), iron (Fe), copper (Cu), and zinc (Zn). Repeated measures statistical analysis of variance (ANOVA) was used. Results showed that (1) the appliances released measurable quantities of all ions examined; (2) the change in pH had a very strong effect (up to 100-fold) on the release of ions; and (3) the release of ions was dependent on wire composition, but it was not proportional to the content of metal in the wire. The largest number of ions was released during the first week of appliance immersion. CONCLUSION: Levels of released ions are sufficient to cause delayed allergic reactions. This must be taken into account when type of archwire is selected, especially in patients with hypersensitivity or compromised oral hygiene. PMID- 19123704 TI - Relationships between dental roots and surrounding tissues for orthodontic miniscrew installation. AB - OBJECTIVE: To elucidate relationships between the dental roots and surrounding tissues in order to prevent complications after placement of a miniscrew. MATERIALS AND METHODS: Twenty human mandibles and maxillas were used for this study. In the 200 sections of each mandible and maxilla, nine items were measured to investigate the relationships between the dental roots. RESULTS: The interroot distance increased from anterior to posterior teeth and from the cervical line to the root apex in both the maxilla and the mandible. In the maxilla, the greatest interroot distance was between the second premolar and the first molar. In the mandible, the greatest interroot distance was between the first and second molars. The maxillary buccolingual bone width exceeded 10 mm from 7 mm (between canine and first premolar), 5 mm (between second premolar and first molar), and 4 mm (between first and second molars) above the cervical line. The mandibular buccolingual bone width exceeded 10 mm from 7 mm (between second premolar and first molar) and 4 mm (between first and second molars) below the cervical line. CONCLUSIONS: The safest zone for placement of a miniscrew in the maxilla was between the second premolar and the first molar, from 6 to 8 mm from the cervical line. The safest zone for placement of a miniscrew in the mandible was between the first and second molars, less than 5 mm from the cervical line. PMID- 19123705 TI - Surface anatomy of the lip elevator muscles for the treatment of gummy smile using botulinum toxin. AB - OBJECTIVE: To propose a safe and reproducible injection point for botulinum toxin A (BTX-A) as a supplementary method for the treatment of gummy smile, as determined by assessment of the morphologic characteristics of three lip elevator muscles. MATERIALS AND METHODS: A total of 50 hemi-faces from 25 adult cadavers (male 13, female 12; ages, 47 to 88 years) were used in this study. Topographic relations and the directions of the lip elevator muscles (ie, levator labii superioris [LLS], levator labii superioris alaeque nasi [LLSAN], and zygomaticus minor [ZMi]), were investigated. Possible injection points were examined through the study of predetermined surface landmarks. RESULTS: The insertion of the LLS was covered partially or entirely by the LLSAN and the ZMi, and the three muscles converged on the area lateral to the ala. The mean angle between the facial midline and each muscle vector was 25.8 +/- 4.8 degrees for the LLS, 55.7 +/- 6.4 degrees for the ZMi, and -20.2 +/- 3.2 degrees for the LLSAN; no significant differences were noted between male and female subjects or between left and right sides. The three vectors passed near a triangular region formed by three surface landmarks. The center of this triangle, named the "Yonsei point", was suggested as an appropriate injection point for BTX-A. The clinical effectiveness of the injection point was demonstrated in selected cases with or without orthodontic treatment. CONCLUSIONS: Under careful case selection, BTX-A may be an effective treatment alternative for patients with excessive gingival display caused by hyperactive lip elevator muscles. PMID- 19123706 TI - Experimentally created nonbalanced occlusion effects on the thickness of the temporomandibular joint disc in rats. AB - OBJECTIVES: To test the hypothesis that experimentally created physiologically nonbalanced occlusion will not affect the thickness of the temporomandibular joint (TMJ) discs in rats. MATERIAL AND METHODS: Twenty-four 8-week-old Sprague Dawley rats were equally divided into a control group that was left untreated and an experimental group where a nonbalanced occlusion was created. Elastic rubber bands, 1 mm in diameter, were inserted and 1 week later were replaced by plastic material between the first and the second molars of the left maxillary and the right mandibular dentitions to move the first molars about 0.8 mm mesially. This created and maintained a physiologically nonbalanced occlusion. The animals were euthanized 8 weeks later, and the TMJ disc thickness was measured on histologically prepared slices using an electronic meter. Two-way univariate analysis of variance was used to compare the groups (alpha level = .05). RESULTS: The intermediate zone was thicker in the experimental group than in the control group (P = .003), but no differences were found between groups regarding the anterior and posterior bands. There were no significant sex-related effects on this observation. CONCLUSION: The hypothesis is rejected. The results indicate that the intermediate zone of rat TMJ disc has the ability to adapt to the alteration of the space between condyle and fossa caused by occlusion changes. Further studies on larger groups that are followed for longer times are needed. PMID- 19123707 TI - Dental arch widths of Southern Chinese. AB - OBJECTIVE: To investigate the dental arch width of the Southern Chinese across buccal cusps, central fossae, or lingual cusps and compare these values with study findings in different populations and using different definitions. MATERIALS AND METHODS: The dental arch dimensions of study casts of an unselected sample from a 12-year-old Hong Kong Oral Health Survey of 12-year-old children (n = 358; 210 boys and 148 girls) were measured. RESULTS: When compared with Caucasians, the Southern Chinese were characterized by a wider dental arch width. However, variations were great. All maxillary and mandibular male arch widths were significantly larger than female arch widths, except at the incisor regions. CONCLUSION: This study yielded a database about dental arch widths by which different studies on these widths can be compared. PMID- 19123708 TI - Preventive effect of ozone on the development of white spot lesions during multibracket appliance therapy. AB - OBJECTIVE: To test the null hypotheses: (1) there is no difference in the caries protective effect of ozone and Cervitec/Fluor Protector during multibracket (MB) appliance therapy, and (2) DIAGNOdent and quantitative light-induced fluorescence (QLF) are not superior to a visual evaluation of initial caries lesions. MATERIALS AND METHODS: Twenty right-handed patients with a very poor oral hygiene who required full MB appliance therapy were analyzed during 26 months. In a split mouth-design, the four quadrants of each patient were either treated with ozone, a combination of Cervitec and Fluor Protector, or served as untreated controls. The visible plaque index (VPI) and white spot formation were analyzed clinically. DIAGNOdent and QLF were used for a quantitative assessment of white spot formation. RESULTS: The average VPI in all four dental arch quadrants amounted to 55.6% and was independent of the preventive measure undertaken. In the quadrants treated with Cervitec/Fluor Protector, only 0.7% of the areas developed new, clinically visible white spots. This was significantly (P < .05) less than in the quadrants treated with ozone (3.2%). The lesions detected with QLF only partially corresponded to the clinically detected white spots, while DIAGNOdent proved to be unable to detect any changes at all. CONCLUSIONS: The caries protective effect of Cervitec/Fluor Protector during MB therapy was superior to ozone, and a visual evaluation of initial caries lesions was superior to both DIAGNOdent and QLF. PMID- 19123709 TI - Shear bond strength of brackets bonded to enamel with a self-etching primer. Effects of increasing storage time after activation. AB - OBJECTIVE: To evaluate bonding efficacy of activated Transbond Plus Self-Etching Primer (TPSEP) used at different time points with Transbond XT to bond metallic orthodontic brackets to bovine incisors. MATERIALS AND METHODS: The inferior incisors of 210 bovines were randomly divided into seven groups (n = 30). TPSEPs were mixed, activated, and kept activated for 30 (group 30), 21 (group 21), 15 (group 15), 7 (group 7), 3 (group 3), or 1 (group 1) days before bonding, and in one group (group 0) TPSEP was used immediately after mixed. At day zero, incisors in each group were bonded in exactly the same way. After applying TPSEP, brackets were bonded with Transbond XT, according to the manufacturer's instructions. After 24 hours, shear bond strength (SBS) tests were performed for all samples at a crosshead speed of 0.5 mm/min, and the Adhesive Remnant Index was scored. RESULTS: There were no significant differences between the SBS of groups 0, 1, 3, 7, and 15 (P > .05) However, those groups had higher SBS (P < .05) compared with groups 21 and 30. No significant difference (P > .05) was observed between groups 21 and 30. Despite the decrease in SBS for groups 21 and 30, bond strength values were still satisfactory. CONCLUSION: After activation, the TPSEP mix can be stored for a period of 15 days without losing its adhesive properties. PMID- 19123710 TI - The relationship between digital model accuracy and time-dependent deformation of alginate impressions. AB - OBJECTIVES: To investigate the effects of different storage periods of alginate impressions on digital model accuracy. MATERIALS AND METHODS: A total of 105 impressions were taken from a master model with three different brands of alginates and were poured into stone models in five different storage periods. In all, 21 stone models were poured and immediately were scanned, and 21 digital models were prepared. The remaining 84 impressions were poured after 1, 2, 3, and 4 days, respectively. Five linear measurements were made by three researchers on the master model, the stone models, and the digital models. Time-dependent deformation of alginate impressions at different storage periods and the accuracy of traditional stone models and digital models were evaluated separately. RESULTS: Both the stone models and the digital models were highly correlated with the master model. Significant deformities in the alginate impressions were noted at different storage periods of 1 to 4 days. Alginate impressions of different brands also showed significant differences between each other on the first, third, and fourth days. CONCLUSIONS: Digital orthodontic models are as reliable as traditional stone models and probably will become the standard for orthodontic clinical use. Storing alginate impressions in sealed plastic bags for up to 4 days caused statistically significant deformation of alginate impressions, but the magnitude of these deformations did not appear to be clinically relevant and had no adverse effect on digital modeling. PMID- 19123711 TI - Friction between archwires of different sizes, cross-section and alloy and brackets ligated with low-friction or conventional ligatures. AB - OBJECTIVE: To test the null hypothesis that no statistically significant difference in frictional resistance is noted when round or rectangular archwires are used in conjunction with low-friction ligatures (small, medium, or large) or conventional ligatures. MATERIALS AND METHODS: A total of 10 stainless steel brackets, a 0.022-in slot, and various orthodontic archwires, ligated with low friction ligatures or conventional ligatures, were tested to compare frictional resistance. The archwires employed were 0.014-in and 0.016-in nickel titanium (NiTi), 0.018-in stainless steel (SS), 0.016 x 0.022-in NiTi, 0.016 x 0.022-in SS, 0.017 x 0.025-in titanium molybdenum alloy (TMA), 0.017 x 0.025-in NiTi, 0.017 x 0.025-in SS, 0.019 x 0.025-in SS, and 0.019 x 0.025-in NiTi. Each bracket/archwire combination was tested 10 times in the dry state at an ambient temperature of 34 degrees C. RESULTS: Low-friction ligatures with round archwires showed statistically significantly lower frictional resistance than did conventional ligatures. When coupled with 0.016 x 0.022-in NiTi and SS, no statistically significant difference was observed among the four groups. When coupled with 0.017 x 0.025-in archwires, low-friction ligatures showed statistically significantly greater frictional resistance than was seen with conventional ligatures. When coupled with 0.019 x 0.025-in NiTi, low-friction ligatures showed statistically significantly greater frictional resistance than did conventional ligatures, but no difference among the four groups was observed with the 0.019 x 0.025-in SS. No significant difference was assessed among low friction ligatures of different sizes. CONCLUSION: Low-friction ligatures show lower friction when compared with conventional ligatures when coupled with round archwires, but not when coupled with rectangular ones. PMID- 19123712 TI - Microleakage under ceramic and metallic brackets bonded with resin-modified glass ionomer. AB - OBJECTIVE: To test the null hypothesis that there is no significant difference between the microleakage of adhesive interferences at the occlusal and gingival margins of both ceramic and metallic brackets bonded with light-cured resin modified glass ionomer and a conventional adhesive. MATERIALS AND METHODS: Sixty freshly extracted human maxillary premolar teeth were randomly divided into four groups of 15 teeth each. Metal and ceramic brackets were bonded to groups 1 and 2 with resin-modified glass ionomer adhesive (RMGIA). Metal and ceramic brackets were bonded to group 3 and group 4 with a conventional adhesive (CA) system. A dye-penetration method was used for microleakage evaluation. Microleakage from the occlusal and gingival margins was determined by a stereomicroscope for the enamel-adhesive and bracket-adhesive interfaces. Statistical analysis was performed using the Kruskal-Wallis test and the Mann-Whitney U-test with a Bonferroni correction. RESULTS: The gingival side of all groups exhibited higher microleakage scores compared with the occlusal side for both adhesive interfaces. All bracket and adhesive combinations displayed statistically significant differences in microleakage between the enamel-adhesive and adhesive-bracket interfaces at the occlusal and gingival sides of the brackets (P < .001). When the adhesive systems were compared, the RMGIA showed more microleakage than the CA between the different interfaces. CONCLUSIONS: The hypothesis is rejected. RMGIA results in more microleakage between enamel-adhesive interfaces. PMID- 19123713 TI - Orthodontic treatment in a patient with cleidocranial dysostosis. AB - Cleidocranial dysostosis is a rare congenital skeletal disorder, associated with clavicular hypoplasia or aplasia, delayed closure of cranial fontanels, brachycephalic skull, delayed exfoliation of primary dentition, eruption of permanent teeth, and multiple supernumerary and morphologic abnormalities of the maxilla and mandible. The disorder is caused by mutation in the CBFA1 gene, on the short arm of chromosome 6p21. The prevalence of cleidocranial dysostosis is estimated one per million, without sex or ethnic group predilection. The purpose of this paper is to describe the orthodontic treatment in a patient with cleidocranial dysostosis. Therapy may include removal of supernumerary teeth, surgical exposure of impacted teeth, and orthodontic treatment. PMID- 19123714 TI - A severe reaction to ni-containing orthodontic appliances. AB - Exposure to nickel-containing orthodontic appliances may cause intra- or extraoral allergic reactions. Nickel is the most typical antigen implicated in causing allergic contact dermatitis, which is a Type IV delayed hypersensitivity immune response. This report presents an unusual reaction to nickel during the orthodontic treatment of an adult female patient. The patient had no previous history of allergy and had been wearing fixed metal upper appliances while in orthodontic treatment to assist the eruption of her impacted teeth. The adverse hypersensitivity reactions appeared only after the surgical exposure and included severe signs of eczematic and urticarial reactions of the face with redness, irritation, itching, eczema, soreness, fissuring, and desquamation as well as intraoral diffuse red zones. Diagnostic patch testing performed by the allergist revealed sensitization to nickel (++++ score). Treatment was achieved with nickel free appliances. PMID- 19123715 TI - Microleakage under orthodontic brackets using high-intensity curing lights. AB - OBJECTIVE: To compare the microleakage of the enamel-adhesive-bracket complex at the occlusal and gingival margins of brackets bonded with high-intensity light curing lights and conventional halogen lights. MATERIALS AND METHODS: Forty-five freshly extracted human maxillary premolar teeth were randomly separated into three groups of 15 teeth each. Stainless steel brackets were bonded in all groups according to the manufacturer's recommendations. Specimens (15 per group) were cured for 40 seconds with a conventional halogen light, 20 seconds with light emitting diode (LED), and 6 seconds with plasma arc curing light (PAC). After curing, the specimens were further sealed with nail varnish, stained with 0.5% basic-fuchsine for 24 hours, sectioned and examined under a stereomicroscope, and scored for microleakage for the enamel-adhesive and bracket-adhesive interfaces from both the occlusal and gingival margins. Statistical analyses were performed using Kruskal-Wallis and Mann-Whitney U-tests with a Bonferroni correction. RESULTS: The type of light curing unit did not significantly affect the amount of microleakage at the gingival or occlusal margins of investigated interfaces (P >.05). The gingival sides in the LED and PAC groups exhibited higher microleakage scores compared with those observed on occlusal sides for the enamel-adhesive and adhesive-bracket interfaces. The halogen light source showed similar microleakage at the gingival and occlusal sides between both adhesive interfaces. CONCLUSIONS: High-intensity curing units did not cause more microleakage than conventional halogen lights. This supports the use of all these curing units in routine orthodontic practice. PMID- 19123716 TI - Analysis of orthodontically induced root resorption using micro-computed tomography (Micro-CT). AB - OBJECTIVE: To establish a protocol for assessing orthodontically induced tooth root resorption (OITRR) in human premolars using micro-computed tomography. MATERIALS AND METHODS: Ten extracted maxillary first premolars were obtained from eight healthy adolescent patients; five of these premolars were extracted before any orthodontic movement was applied to them, and the other five premolars were involved in nonextraction orthodontic treatment for 1 year before treatment plan modification lead to extraction treatment. Using reconstructed scanned images, we measured several key resorption lacunae parameters, including the number, volume, and depth on each surface, as well as its extension coronoapically. RESULTS: Orthodontic treatment resulted in a significantly greater tooth root resorption lacunae volume, number, maximum depth, and coronoapical extension as well as in areas of dentin demineralization subjacent to the resorption lacunae than in normal premolars. CONCLUSIONS: We have established a protocol to evaluate OITRR quantitatively and have provided a method to predict further resorption based on dentine demineralization. PMID- 19123717 TI - Bond strength of amorphous calcium phosphate-containing orthodontic composite used as a lingual retainer adhesive. AB - OBJECTIVE: To evaluate the shear bond strength and fracture mode difference between amorphous calcium phosphate (ACP)-containing adhesive and conventional resin-based composite material used as an orthodontic lingual retainer adhesive. MATERIALS AND METHODS: Forty crowns of extracted lower human incisors were mounted in acrylic resin, leaving the buccal surface of the crowns parallel to the base of the molds. The teeth were randomly divided into two groups: experimental and control, containing 20 teeth each. Conventional lingual retainer composite (Transbond-LR, 3M-Unitek) and ACP-containing orthodontic adhesive (Aegis-Ortho) were applied to the teeth surface by packing the material into the cylindrical plastic matrices with a 2.34-mm internal diameter and a 3-mm height (Ultradent) to simulate the lingual retainer bonding. For shear bond testing, the specimens were mounted in a universal testing machine, and an apparatus (Ultradent) attached to a compression load cell was applied to each specimen until failure occurred. The shear bond data were analyzed using Student's t-test. Fracture modes were analyzed by chi(2) test. RESULTS: The statistical test showed that the bond strengths of group 1 (control Transbond-LR, mean: 24.77 +/- 9.25 MPa) and group 2 (ACP-containing adhesive, mean: 8.49 +/- 2.53 MPa) were significantly different from each other. In general, a greater percentage of the fractures were adhesive at the tooth-composite interface (60% in group 1 and 55% in group 2), and no statistically significant difference was found between groups. CONCLUSION: The ACP-containing Aegis-Ortho adhesive resulted in a significant decrease in bond strength to the etched enamel surface. PMID- 19123718 TI - Pain experience during initial alignment with a self-ligating and a conventional fixed orthodontic appliance system. A randomized controlled clinical trial. AB - OBJECTIVES: To test the hypotheses that (1) there is no difference in the pain experience during the week following initial placement of two orthodontic appliances (SmartClip and Victory; 3M Unitek, Monrovia, Calif); and (2) there is no difference in the pain experience during removal and insertion of orthodontic archwires with these brackets. MATERIALS AND METHODS: Sixty-six consecutive patients were treated with a self-ligating bracket system (SmartClip) or a conventional appliance (Victory) on the basis of computer-generated random allocation. After appliance placement and engagement of a 0.016'' nickel-titanium archwire, pain experience was recorded after 4, 24, and 72 hours and after 7 days with the use of a visual analog system (VAS) questionnaire. At a subsequent visit, participants documented pain experiences during removal and insertion of 0.019 x 0.025'' archwires on an additional 100 mm VAS questionnaire. Independent t-tests and analyses of covariance were used to analyze normally distributed data; the Mann-Whitney U-test was used for skewed distributions. RESULTS: Forty eight (72.2%) and fifty-one (77.3%) subjects completed the first and second parts of the study, respectively. Bracket type had no influence on pain experience at 4 hours (P = .958), 24 hours (P = .289), 72 hours (P = .569), and 7 days (P = .756) following appliance placement. However, bracket type significantly influenced pain experience during archwire removal (P = .001) and insertion (P = .013). CONCLUSIONS: Hypothesis 1 cannot be rejected. The bracket type had no effect on subjective pain experience during the first week after initial placement of two preadjusted orthodontic appliances. Hypothesis 2 was rejected. Significantly greater discomfort was experienced during archwire insertion and removal with the SmartClip appliance. PMID- 19123719 TI - Linear accuracy of cone beam CT derived 3D images. AB - OBJECTIVE: To compare the in vitro reliability and accuracy of linear measurements between cephalometric landmarks on cone beam computed tomography (CBCT) 3D volumetric images with varying basis projection images to direct measurements on human skulls. MATERIALS AND METHODS: Sixteen linear dimensions between 24 anatomic sites marked on 19 human skulls were directly measured. The skulls were imaged with CBCT (i-CAT, Imaging Sciences International, Hatfield, Pa) at three settings: (a) 153 projections, (b) 306 projections, and (c) 612 projections. The mean absolute error and modality mean (+/- SD) of linear measurements between landmarks on volumetric renderings were compared to the anatomic truth using repeated measures general linear model (P < or = .05). RESULTS: No difference in mean absolute error between the scan settings was found for almost all measurements. The average skull absolute error between marked reference points was less than the distances between unmarked reference sites. CBCT resulted in lower measurements for nine dimensions (mean difference range: 3.1 mm +/- 0.12 mm to 0.56 mm +/- 0.07 mm) and a greater measurement for one dimension (mean difference 3.3 mm +/- 0.12 mm). No differences were detected between CBCT scan sequences. CONCLUSIONS: CBCT measurements were consistent between scan sequences and for direct measurements between marked reference points. Reducing the number of projections for 3D reconstruction did not lead to reduced dimensional accuracy and potentially provides reduced patient radiation exposure. Because the fiducial landmarks on the skulls were not radio-opaque, the inaccuracies found in measurement could be due to the methods applied rather than to innate inaccuracies in the CBCT scan reconstructions or 3D software employed. PMID- 19123720 TI - Hard and soft tissue stability of orthognathic surgery. AB - OBJECTIVE: To test the hypothesis that there is no difference in the stability of the hard and soft tissue changes following a surgical mandibular setback using a sagittal split ramus osteotomy or an intraoral vertical ramus osteotomy. MATERIALS AND METHODS: The samples consisted of 45 female patients with mandibular prognathism, who were divided into two groups. Twenty-three underwent a sagittal split ramus osteotomy (SSRO) with rigid fixation by titanium mini screws and maxillomandibular fixation (MMF) for 14.0 +/- 0.7 days. Twenty-two underwent intraoral vertical ramus osteotomy (IVRO) without rigid fixation, and MMF was released 21.5 +/- 3.3 days after surgery. Data were collected from standardized cephalometric radiographs taken at the start of preoperative orthodontic treatment (T1), immediately after surgery (MMF, T2), and the completion of postoperative treatment (T3). Angular measurements were compared on each of T1, T2, and T3. RESULTS: There were no significant differences in the pretreatment hard and soft tissue morphology between the SSRO and IVRO. However, when fixation was released, the mandible was posteriorly positioned in the IVRO group. In the soft tissue profile, the mental region was located backward in the IVRO group at postoperative stage (T3). CONCLUSIONS: The hypothesis is rejected. The soft tissue profile of the IVRO group especially showed a retromandibular position after postoperative treatment in comparison with the SSRO group. This tendency of the IVRO group would contribute to the database for treatment planning and prediction. PMID- 19123721 TI - Soft-tissue facial characteristics of attractive Italian women as compared to normal women. AB - OBJECTIVE: To compare the facial characteristics of two different groups of attractive women with those of reference women. MATERIALS AND METHODS: The three dimensional coordinates of 50 facial landmarks were collected in 71 healthy reference women (18-30 years old) and in 24 coetaneous "attractive" women selected during two different beauty competitions; soft tissue facial angles, distances, areas, and volumes were computed and compared using analysis of variance. RESULTS: When compared with reference women, both groups of attractive women shared several similar facial characteristics: relatively large forehead (P < .001), reduced mandible (P = .008), and rounded face (reduced surface-to-volume ratio, P = .002). They had a more acute soft tissue profile, an increased upper facial width (P < .001) and middle facial depth, larger mouth, and more voluminous lips (P = .005) than reference women. CONCLUSIONS: Both groups of attractive women had several facial characteristics suggesting babyness. Nonetheless, each group of women was characterized by a different development of these features. Esthetic reference values can be a useful tool for clinicians, but should always consider the characteristics of individual faces. PMID- 19123722 TI - Intracranial lesions mimicking neoplasms. AB - CONTEXT: A broad spectrum of nonneoplastic conditions can mimic a brain tumor, both clinically and radiologically. In this review we consider these, taking into consideration the following etiologic categories: infection, demyelination, vascular diseases, noninfectious inflammatory disorders, and iatrogenic conditions. We give an overview of such diseases, which represent a potential pitfall for pathologists and other clinicians involved in patient care, and present selected cases from each category. OBJECTIVE: To illustrate the radiologic and pathologic features of nontumoral intracranial lesions that can clinically and radiologically mimic neoplasia. DATA SOURCES: Case-derived material and literature review. CONCLUSIONS: A variety of nonneoplastic lesions can present clinically and radiologically as primary or metastatic central nervous system tumors and result in surgical biopsy or resection of the lesion. In such situations, the pathologist has an important role to play in correctly determining the nature of these lesions. Awareness of the entities that can present in this way will assist the pathologist in the correct diagnosis of these lesions. PMID- 19123723 TI - Glucose harmonization: the role of accurate calibrators. PMID- 19123724 TI - How does a pathologist make a diagnosis? AB - A pathologic diagnosis is the result of a complex series of activities, mastered by the pathologist. The nature of these activities is, however, rarely talked about in depth. The medical literature occasionally discusses aspects of the pathologic diagnosis processes, generally departing from the pathologic practice. The lack of a model makes discussions about the subject a matter of preference or personal style. Educational programs are largely based on the apprenticeship model, and the development of specific abilities rests on the personal aspects of both apprentice and mentor. A model for the pathologic diagnostic process is proposed. The process of diagnosis can be viewed as an action plan, encompassing 4 domains: (1) cognitive, (2) communicative, (3) normative, and (4) medical conduct. The cognitive domain involves processes of perception, attention, memory, search, hypothesis creation, and verification, among others. Communicative skills consist of providing arguments in support of a diagnostic conclusion, with adequate clinical and relevant pathologic information. Pathologic diagnosis is also subject to technical rules (based on empirical experiences), rules of rational choice (strategies aiming at definite goals), and consensual rules among peers. Finally, the pathologic diagnosis has to be evaluated in the sphere of medical conduct, from the perspectives of both the pathologist and the referring clinician. An understanding of the diagnostic process from a theoretic perspective will benefit pathology as a science and a medical specialty because it provides the basis for understanding diagnostic variations and discrepancies. Pathologic difficulties or errors can be mapped, allowing the institution of specific remedies. This model may also enhance training and educational strategies because specific emphasis can be directed toward a particular difficulty. PMID- 19123725 TI - College of American Pathologists protocol for the reporting of ductal carcinoma in situ. PMID- 19123726 TI - Primary intestinal intraepithelial natural killer-like T-cell lymphoma: case report of a distinct clinicopathologic entity. AB - Intestinal T-cell lymphoma is a heterogenous group. These tumors differ in their association with enteropathy, intraepithelial or nonintraepithelial origin, primary or secondary involvement, and T-cell or natural killer-like T-cell immunophenotype. There are also nonneoplastic conditions, such as celiac disease, refractory sprue, and reactive T-cell infiltration that mimic intestinal T-cell lymphoma. Therefore, the differential diagnosis requires extensive morphologic, immunophenotypic, and molecular genetic studies. A subset of primary intestinal intraepithelial T-cell lymphoma has emerged in recent years that is distinguished from enteropathy-type T-cell lymphoma in terms of clinical presentation (nonenteropathic), morphology (monomorphic small to medium-sized cells), immunophenotype (CD3(-)CD8(+)CD56(+)), and cytogenetics. We report such a case with a unique immunophenotype (CD3(-), cytoplasmic CD3(+), CD4(-), CD8(+), CD5( ), CD7(+), CD16(-), CD56(+), CD57(-), CD103(+), T-cell intracellular antigen 1(+), and betaF1(+)) that is comparable to that of a newly identified subset of intraepithelial T cells. The tumor progressed rapidly and the patient died within 6 months after the onset of the disease. We recommend a large monoclonal antibody panel for the differential diagnosis of this heterogeneous group of T-cell lymphoma. PMID- 19123727 TI - Resolution of giant cell myocarditis after extended ventricular assistance. AB - Lower recurrence rates and improved long-term outcomes are the goal of treatment for giant cell myocarditis (GCM). We describe a case of GCM in an Asian woman who presented with new onset palpitations and syncope. She initially had normal systolic function by echocardiography with magnetic resonance imaging evidence of infiltrative cardiomyopathy. She underwent implantation of a biventricular assist device (BiVAD) because of rapidly deteriorating hemodynamic status. Giant cell myocarditis was diagnosed at that time by surgical biopsy. She was not treated with immunosuppressive therapy because of low likelihood of recovery and concerns for potential infection with the BiVAD in place. She received a heart transplant 12 months later and had extensive fibrosis but no evidence of active GCM in the heart explant. The role of extended BiVAD support in patients with GCM should be further investigated. PMID- 19123728 TI - Pediatric follicular lymphoma: a rare clinicopathologic entity. AB - Follicular lymphoma, although common in adults, is rare in children. Pediatric follicular lymphoma has a more favorable prognosis than adult follicular lymphoma, even though it is often of higher grade. Children with follicular lymphomas are generally at a lower clinical stage, respond well to less aggressive therapy, and have a better survival than adults. Follicular lymphoma must be distinguished from reactive follicular hyperplasia, which it may mimic. Immunohistochemical and molecular markers serve to facilitate this distinction, as well as careful attention to clinical and morphologic details. It is important to recognize pediatric follicular lymphoma as a unique clinicopathologic entity to properly diagnose and manage these patients. It may represent a subset of follicular lymphoma with a particularly good prognosis. PMID- 19123729 TI - Hamartoma of the spleen. AB - Splenic hamartoma is a rare, benign vascular proliferation that is often found incidentally while working up other complaints or at autopsy. Women more commonly present with symptoms related to mass effect than men. Histologic findings consist of unorganized vascular channels of varying width, with intervening red pulp-like disorganized stroma with or without lymphoid follicles. The endothelial cells are similar to those of normal splenic sinuses. Although rendering a diagnosis can be difficult, endothelial cells that are positive for CD8 are a key feature that differentiate hamartoma from other vascular lesions of the spleen. Clinical, radiologic, and histologic correlation is essential to ensure this benign lesion is not mistaken for malignancy. PMID- 19123730 TI - Protocol for the examination of specimens from patients with ductal carcinoma in situ of the breast. PMID- 19123732 TI - Comparative analysis of size estimation by mapping and counting number of blocks with ductal carcinoma in situ in breast excision specimens. AB - CONTEXT: The size of ductal carcinoma in situ (DCIS) is a significant predictor of local tumor recurrence and is used for selection of patients for conservative versus aggressive therapy. A standardized method for size assessment is lacking. OBJECTIVE: To evaluate 2 commonly used methods for measurement of DCIS extent: one based on the distribution of the lesion in sequential series of sections (mapping method) and the other on the number of sections with DCIS (block method). DESIGN: Ninety-eight consecutive cases of DCIS, measuring at least 1.0 cm, were retrieved from our files. All specimens were serially sectioned along the long axis. The size of DCIS was calculated for each case by 2 different methods: (1) mapping method (average thickness of each slice x number of consecutive slices with DCIS) and (2) block method (number of blocks with DCIS x 0.3 cm). Additional calculations were performed by using 0.35, 0.4, and 0.5 cm as multiplication factors for the block method in order to improve concordance. RESULTS: The block method underestimated the size in 71 cases (72%) by 4.5% to 81.3% (mean, 33%). Using 0.4 cm as the multiplication factor improved concordance, while multiplying by 0.5 cm led to an overestimation of size. CONCLUSIONS: Assessment of DCIS size by the block method is inaccurate and underestimates size in most cases (72%), with an average reduction of 33%. Using 0.4 cm as the multiplication factor improves concordance. A standardized method for size estimation is necessary for effective patient management. PMID- 19123733 TI - Measuring extent of ductal carcinoma in situ in breast excision specimens: a comparison of 4 methods. AB - CONTEXT: Measuring the extent of nonpalpable ductal carcinoma in situ (DCIS) in a breast specimen is challenging but important because it influences patient management. There is no standardized method for estimating the extent of DCIS, although serial sequential sampling with mammographic correlation is considered an accurate method. OBJECTIVE: To estimate the extent of DCIS using various methods and to compare these estimations with the extent as determined by the serial sequential sampling method. DESIGN: A total of 78 primary breast excisions with DCIS were retrospectively reviewed. All specimens had been sampled using the serial sequential sampling method, which involved mapping the location of each block on the sliced specimen radiograph and calculating the extent through 3 dimensional reconstruction. The other measures for estimating extent included (1) calculating size based on areas of calcification, (2) recording the number of blocks involved by DCIS and multiplying that number by 0.3 cm, and (3) measuring the largest extent of DCIS on a single slide. RESULTS: All 3 alternative methods tended to underestimate the DCIS. Discrepancies became more pronounced as size increased. The percentage of cases estimated to within 1 cm of the serial sequential sampling method were 81%, 72%, and 50%, respectively, for the calcification, blocks, and single-slide methods; differences of more than 2 cm were seen in 9%, 8%, and 30% of cases, respectively. CONCLUSIONS: The single slide method performed poorly and should be used only when DCIS is limited to a single slide. Although the calcification and the blocks methods gave better estimates, both produced substantial underestimates and/or overestimates that could affect clinical decision making. PMID- 19123734 TI - Physician satisfaction with clinical laboratory services: a College of American Pathologists Q-probes study of 138 institutions. AB - CONTEXT: Monitoring customer satisfaction is a valuable component of a laboratory quality improvement program. OBJECTIVE: To survey the level of physician satisfaction with hospital clinical laboratory services. DESIGN: Participating institutions provided demographic and practice information and survey results of physician satisfaction with defined aspects of clinical laboratory services, rated on a scale of 1 (poor) to 5 (excellent). RESULTS: One hundred thirty-eight institutions participated in this study and submitted a total of 4329 physician surveys. The overall satisfaction score for all institutions ranged from 2.9 to 5.0. The median overall score for all participants was 4.1 (10th percentile, 3.6; 90th percentile, 4.5). Physicians were most satisfied with the quality/reliability of results and staff courtesy, with median values of excellent or good ratings of 89.9%. Of the 5 service categories that received the lowest percentage values of excellent/good ratings (combined scores of 4 and 5), 4 were related to turnaround time for inpatient stat, outpatient stat, routine, and esoteric tests. Surveys from half of the participating laboratories reported that 96% to 100% of physicians would recommend the laboratory to other physicians. The category most frequently selected as the most important category of laboratory services was quality/reliability of results (31.7%). CONCLUSIONS: There continues to be a high level of physician satisfaction and loyalty with clinical laboratory services. Test turnaround times are persistent categories of dissatisfaction and present opportunities for improvement. PMID- 19123735 TI - Appeals in gynecologic cytology proficiency testing: review and analysis of data from the 2006 College of American Pathologists gynecologic cytology proficiency testing program. AB - CONTEXT: In 2006, 9643 participants took the initial College of American Pathologists (CAP) Proficiency Test (PT). Failing participants may appeal results on specific test slides. Appeals are granted if 3 referee pathologists do not unanimously agree on the initial reference diagnosis in a masked review process. OBJECTIVES: To investigate causes of PT failures, subsequent appeals, and appeal successes in 2006. DESIGN: Appeals were examined, including patient demographic information, Centers for Medicare and Medicaid Services category (A, B, C, or D), exact reference diagnosis, examinees per appeal, examinee's Centers for Medicare and Medicaid Services category, referee's Centers for Medicare and Medicaid Services category, slide preparation type, and slide field validation rate. RESULTS: There was a 94% passing rate for 2006. One hundred fifty-five examinees (1.6%) appealed 86 slides of all preparation types. Forty-five appeals (29%) were granted on 21 slides; 110 appeals (72%) were denied on 65 slides. Reference category D and B slides were most often appealed. The highest percentage of granted appeals occurred in category D (35% slides; 42% of participants) and the lowest occurred in category B (9% slides; 8% of participants). The field validation rate of all appealed slides was greater than 90%. CONCLUSIONS: Despite rigorous field validation of slides, 6% of participants failed. Thirty percent of failing participants appealed; most appeals involved misinterpretation of category D as category B. Referees were never unanimous in their agreement with the participant. The participants and referees struggled with the reliability and reproducibility of finding rare cells, "overdiagnosis" of benign changes, and assigning the morphologically dynamic biologic changes of squamous intraepithelial lesions to static categories. PMID- 19123736 TI - Flow cytometric immunophenotyping of anaplastic large cell lymphoma. AB - CONTEXT: Anaplastic large cell lymphoma (ALCL) is usually diagnosed by histologic and immunohistochemical analysis. However, fine-needle aspiration is becoming a popular alternative to lymph node biopsy, and flow-cytometric immunophenotyping is often used to analyze fine-needle aspiration specimens. OBJECTIVE: To review our experience using flow-cytometric immunophenotyping to assess cases of ALCL and to evaluate the diagnostic utility of this technique. DESIGN: Each case of ALCL was assessed by flow cytometry with 3-color or 4-color antibody panels, and data were reanalyzed by cluster analysis using Paint-a-Gate for cases with retrievable flow cytometry data files. Anaplastic lymphoma kinase (ALK) was assessed by using immunohistochemistry. RESULTS: Twenty-three ALCL cases were analyzed by flow cytometry. In 4 cases, neoplastic cells could not be identified. In the remaining 19 cases (11 ALK(+), 8 ALK(-)), all were positive for CD30 and CD45. Anaplastic large cell lymphoma cells were large and usually CD45 bright, with many or most cells falling in the region of monocytes on the CD45/side scatter plot. The frequencies of T-cell antigen expression in ALK(+) cases were CD2, 67%; CD7, 60%; CD3, 45%; CD4, 33%; CD5, 14%; and CD8, 14%. In ALK(-) cases, the frequencies of the T-cell antigen expression were CD2, 100%; CD3, 50%; CD4, 40%; CD7, 40%; CD5, 25%; and CD8, 20%. CONCLUSIONS: Flow cytometry can be used to immunophenotype ALCL cases. Neoplastic cells may be few, and they may fall outside of the lymphocyte gate. Cluster analysis using software like Paint-A-Gate is often helpful because it allows for flexible, sequential gating strategies to identify and characterize the neoplastic cells. PMID- 19123737 TI - Impact of digital image manipulation in cytology. AB - CONTEXT: Digital images have become an important component of cytology practice. They are used in telecytology, automated screening, educational material, and Web sites and have potential for use in proficiency testing. However, there has been no formal evaluation to date to determine if digital image manipulation (intentional or unintentional) can affect their interpretation. OBJECTIVE: To investigate whether alteration of digital cytology images affects diagnosis. DESIGN: Acquired digital images of ThinPrep Papanicolaou test slides were manipulated (rotated 90 degrees and brightness, contrast, red-green-blue color, and luminosity adjusted) using Photoshop. A test composed of these altered images, along with their original (unaltered) image and exact duplicates was given to 22 cytologists (13 cytotechnologists, 8 cytopathologists, and 1 fellow). All images were rated as negative, atypical (atypical squamous cells of undetermined significance), low-grade squamous intraepithelial lesion, high-grade squamous intraepithelial lesion, or positive for cancer. Weighted kappa and heterogeneity chi(2) statistics were used to measure levels of agreement and assess concordance between groups. RESULTS: The level of agreement for identical duplicate images was excellent (kappa = 0.81), compared with the poor agreement for manipulated image pairs (kappa = 0.21), a statistically significant difference (P < .001). For all altered image types agreement was poor. There was no significant difference between cytotechnologists and cytopathologists in level of agreement (P = .56). CONCLUSIONS: Manipulation of a Papanicolaou test digital image, irrespective of the specific category of cytologic material photographed, significantly affects its interpretation by both cytotechnologists and cytopathologists. This suggests that care needs to be taken when digital cytology images are used, to specifically ensure that their alteration does not affect diagnosis. PMID- 19123738 TI - Micropapillary variant of urothelial carcinoma in the upper urinary tract: a clinicopathologic study of 11 cases. AB - CONTEXT: Micropapillary urothelial carcinoma (MPUC) is a rare variant of urothelial carcinoma. Most studies of MPUC have focused on the urinary bladder, but MPUC of the upper urinary tract remains to be investigated. OBJECTIVE: To investigate the pathologic features and clinical significance of MPUC in the upper urinary tract. DESIGN: We searched the pathology files at our institution and identified 11 cases of MPUC of the upper urinary tract. The histology slides were reviewed, and the clinical information was obtained by review of medical charts. RESULTS: The average age of the patients was 64.2 years (range, 22-76 years). The tumors were located in the renal pelvis (n = 5), ureter (n = 4), and ureteropelvic junction (n = 2). In all cases, MPUC accounted for an average of 45% (range, 10%-80%) of the tumor and was associated with conventional urothelial carcinoma. Lymphovascular invasion was present in all cases, and metastasis to lymph node was present in 4 of 5 patients whose lymph nodes were dissected. Two patients presented with pT2 disease, and both were alive without evidence of disease at 85 and 119 months after surgery. The other 9 patients presented with pT3 or pT4 disease: 4 of them died of disease at an average of 18 months; 4 surviving patients developed distant metastases; and 1 surviving patient with limited follow-up (6 months) showed no evidence of disease. CONCLUSIONS: Micropapillary urothelial carcinoma of the upper urinary tract often presents at an advanced stage with lymphovascular invasion and distant metastasis. The presence of MPUC, even focal, indicates a poor clinical course. PMID- 19123739 TI - Carcinomas of the pancreas, gallbladder, extrahepatic bile ducts, and ampulla of vater share a field for carcinogenesis: a population-based study. AB - CONTEXT: Carcinomas co-occur in the pancreas, extrahepatic bile ducts, and ampulla of Vater. We investigated whether cancers originating in these sites represent a field effect similar to that observed in the lung and upper aerodigestive tract. OBJECTIVE: To determine whether a field effect for carcinogenesis exists in the ampulla of Vater, extrahepatic bile ducts, gallbladder, and pancreas. DESIGN: Data were obtained from National Cancer Institute's Surveillance Epidemiology and End Results Program from 1973 through 2005. Cases were compared by age frequency density plots, age-specific incidence rates, and logarithmic plots of the age-specific incidence rates and age of diagnosis. RESULTS: Incidence rates were 11.71, 1.43, 0.88, and 0.49 per 100,000 persons at risk for pancreatic, gallbladder, extrahepatic bile ducts, and ampullary carcinomas, respectively. Age frequency density plots were congruent for cancers originating in all 4 sites. Logarithmic plots of the age-specific incidence rates with age of diagnosis produced parallel linear rate patterns for the 4 sites indicative of similar populations for tumor development. However, density and logarithmic plots of pancreatic endocrine carcinomas, a tumor of different cellular differentiation and carcinogenic pathway, served as a comparison. The endocrine carcinomas showed a different age distribution and nonparallel rate patterns with ductal carcinomas. CONCLUSIONS: Carcinomas of the pancreas, gallbladder, extrahepatic bile ducts, and ampulla have a common embryonic cellular ancestry, differentiation pathways, mucosal histologic patterns, and population-related tumor development indicating a field effect in carcinogenesis. Parallel linear rate patterns indicate (1) the rate of cancer development is similar in all 4 sites even though the absolute incidence rates vary and (2) regardless of location, the ductal epithelium is equally susceptible to malignant transformation. If carcinogenic pathways to cancer are similar, then the different incidence rates seen clinically may depend on the relative surface area of the ductal system in these sites. Pancreatic cancers are most common because the surface area of the pancreas' ductal system is greater than that of the gallbladder, extrahepatic bile ducts, and ampulla. PMID- 19123740 TI - Improvement of coagulation laboratory practice in Thailand: the first-year experience of the national external quality assessment scheme for blood coagulation. AB - CONTEXT: In Thailand until 2005 there had been no external quality assessment scheme at the national level for blood coagulation tests. Only a few laboratories had an external quality assessment for these tests. In the year 2005, the Thailand National External Quality Assessment Scheme for Blood Coagulation was founded. OBJECTIVES: To describe the establishment of the Thailand National External Quality Assessment Scheme for Blood Coagulation (including problems encountered and solutions), its progression and expansion, and the improvement of coagulation laboratory practice in Thailand during 2 trial surveys and 4 formal surveys conducted in the first 1 1/2 years. DESIGN: Between 2005 and 2006, the external quality assessment samples for prothrombin time/international normalized ratio and activated partial thromboplastin time were distributed to the participants as well as the instructions and suggestions for the improvement of laboratory practice. From the data collected, the all-method coefficient of variation of the international normalized ratio and activated partial thromboplastin time was calculated for each survey. RESULTS: The number of participants increased during the first 1 1/2 years that the surveys were conducted, from 109 to 127. Survey data demonstrate an improvement in response rate and an increase in the number of laboratories that determine their own reference ranges and repeat this for every change of reagent lot, using the appropriate anticoagulant. The increased precision of tests is indicated by the decrease of the all-method coefficient of variation of the international normalized ratio and activated partial thromboplastin time. Examples of individual laboratory improvement through feedback are also described. CONCLUSIONS: The improvement of coagulation laboratory practice both through the instructions provided and liaison with participants was observed during the course of this scheme. PMID- 19123741 TI - The autopsy: knowledge, attitude, and perceptions of doctors and relatives of the deceased. AB - CONTEXT: Despite the crucial role that autopsy plays in the development of the science and practice of medicine, autopsy rates have been declining throughout the world in recent decades. OBJECTIVE: To identify factors influencing the acceptance of autopsies in Nigeria. DESIGN: Cross-sectional survey of the knowledge, attitude, and perceptions of doctors and relatives of deceased patients on factors influencing acceptance of autopsy at the University College Hospital, Ibadan, Nigeria. RESULTS: Only 38% of relatives had satisfactory knowledge about autopsy and about 50% of doctors knew that physicians' desire for autopsy should not override patients' consent. Doctors identified difficulty in obtaining consent from relatives of deceased patients, administrative problems in requesting autopsy, and delay in obtaining autopsy report as major reasons responsible for the decline in autopsy requests, whereas relatives of the deceased cited fear of mutilation, concerns about delaying the funeral, and objection by the patient before death as reasons for refusal to grant permission for an autopsy. Sociodemographic factors such as age, occupation, religion, ethnicity, and level of education significantly influenced willingness to give consent for autopsy. CONCLUSIONS: There is need for concerted effort on the part of clinicians, pathologists, the public, and the government to resuscitate and sustain the practice of autopsy in Nigeria. PMID- 19123742 TI - Can GEWF solution improve the retrieval of lymph nodes from colorectal cancer resections? AB - CONTEXT: Lymph node status is an important prognostic factor in colorectal cancer staging. OBJECTIVE: To determine whether the use of GEWF solution (glacial acetic acid, ethanol, distilled water, formaldehyde) increases the yield of lymph nodes (LNs) from colorectal cancer specimens. DESIGN: A prospective study investigated the usefulness of GEWF in identifying LN from colorectal cancer resection specimens. After standard formalin fixation, approximately half of the specimens were placed in GEWF solution, and the remainder were used as a control. The number of LNs, patient sex, length of specimen, site of the specimen, tumor staging, and the number of positive LNs were analyzed. The numbers of retrieved and metastasis-positive LNs were recorded and compared with specimens from the previous year, in which no GEWF solution was used. Residents were educated on the importance of LN status before the study. Data were analyzed using independent sample t tests. The residents also completed a subjective survey at the conclusion of the study. RESULTS: A total of 85 specimens, 45 (53%) with GEWF and 40 (47%) without GEWF, were examined by 11 residents. The mean number of retrieved LNs using GEWF was slightly higher than the control group (19.96 vs 18.30); however, there was no statistical significance (P = .53). The mean number of LNs from the current study (2004- 2005) is significantly higher than that from the previous year (2003-2004) (19.17 vs 11.51, P < .001). CONCLUSIONS: GEWF did not increase the yield of LNs from colorectal cancer specimens. However, application of the guideline for a minimum of 12 LNs significantly increased the LN retrieval. PMID- 19123743 TI - Urine mercury excretion following meso-dimercaptosuccinic acid challenge in fish eaters. AB - CONTEXT: Public awareness of methylmercury in fish has caused patients to seek testing for mercury poisoning. In some patients, the diagnosis of mercury poisoning has been made based on urine mercury excretions following oral dosing of meso-dimercaptosuccinic acid (DMSA), a metal chelator. However, studies comparing urine mercury excretion following DMSA in healthy non-fish eaters with healthy fish eaters could not be located. OBJECTIVES: To describe urinary mercury excretion before and after DMSA in healthy fish eaters and non-fish eaters, and to determine whether urine mercury excretion after DMSA would rise above baseline levels to a greater extent in fish eaters. DESIGN: A total of 24 healthy physicians were assigned to 1 of 3 groups based on fish consumption: non-fish eaters; 1 to 2 fish servings per week; and 3 or more servings per week. Blood mercury concentrations and 12-hour urine mercury and creatinine excretions were measured before and after oral ingestion of 30 mg of DMSA per kilogram of body weight. RESULTS: A total of 24 subjects completed the study, and 2 subsequently were excluded. No difference in baseline urinary mercury excretion was detected between groups. All groups demonstrated an increase in urinary mercury excretion following DMSA, which was higher in fish eaters (P = .04). Multiple linear regression found that the best predictor of a rise in urine mercury excretion following DMSA challenge was the prechelation blood mercury concentration. CONCLUSIONS: In this study of healthy physicians, oral DMSA produced a rise in urine mercury excretion both in non-fish eaters and fish eaters. The increase in chelated mercury excretion was higher in fish eaters. A simple rise in chelated mercury excretion over baseline excretion is not a reliable diagnostic indicator of mercury poisoning. PMID- 19123744 TI - Optimal use of Myco/F lytic and standard BACTEC blood culture bottles for detection of yeast and mycobacteria. AB - CONTEXT: The optimal use of dedicated fungal and mycobacterial blood culture bottles, such as the BACTEC Myco/F Lytic bottle, has not been well defined in clinical practice. OBJECTIVES: To compare the performance of Myco/F Lytic and standard blood culture in clinical practice in an urban tertiary care hospital setting and to implement a strategy for optimal use of Myco/F Lytic culture. DATA SOURCES: Retrospective review of laboratory records. RESULTS: Myco/F Lytic culture did not increase detection of yeasts. Nor did it decrease time to detection except for Candida glabrata, where mean time to positivity dropped from 2.6 +/- 1.1 days in standard to 1.8 +/- 0.8 days in Myco/F Lytic culture. Therefore, an algorithm was developed in which Myco/F Lytic culture was reserved primarily for detection of mycobacteria in patients with severely depressed CD4 counts. Implementation of this algorithm led to a sustained 3-fold reduction in Myco/F Lytic blood culture usage. CONCLUSIONS: Retrospective analysis suggests substantial clinical equivalence of standard blood and Myco/F Lytic culture for detection of yeast. A multifaceted educational approach based on this data led to a sustained change in physician ordering practices and more cost-effective use of resources. PMID- 19123745 TI - In situ detection of inflammatory cytokines and apoptosis in pemphigus foliaceus patients. AB - CONTEXT: Endemic pemphigus foliaceus, or fogo selvagem, is a chronic autoimmune disease characterized by the formation of intraepidermal blisters that reduce adhesion between keratinocytes. Endemic pemphigus foliaceus is associated with the presence of autoantibodies and high levels of cytokines involved in the inflammatory response. OBJECTIVES: To evaluate the expression of the proinflammatory cytokines interleukin 1, interferon gamma, and tumor necrosis factor alpha; the proapoptotic inducers Fas and inducible nitric oxide synthase; and the apoptosis inhibitor Bcl-2; and to evaluate the presence of apoptosis. DESIGN: Skin biopsies from 13 patients with endemic pemphigus foliaceus and controls were evaluated by immunohistochemistry and apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. RESULTS: Proinflammatory cytokines were only detected in cells of the inflammatory exudate. Inducible nitric oxide synthase, Fas, and Bcl-2 were expressed by both epithelial and inflammatory cells. Epithelial apoptosis was observed in 12 cases (92.3%), and subepithelial apoptosis in 11 cases (85%). CONCLUSIONS: This study suggests that apoptosis as well as the local production of proinflammatory cytokines are associated with endemic pemphigus foliaceus lesions. These results may contribute to the development of new therapeutic approaches to endemic pemphigus foliaceus. PMID- 19123746 TI - Altruism born of suffering: the roots of caring and helping after victimization and other trauma. AB - Research on altruism has focused on its positive roots, whereas research on the effects of victimization and suffering has focused on aggression and difficulties in functioning. However, anecdotal evidence, case studies, and some empirical research indicate that victimization and suffering can also lead people to care about and help others. This article examines the relation of "altruism born of suffering" to resilience and posttraumatic growth, and proposes potentially facilitating influences on altruism born of suffering during, after, and preceding victimization and trauma. These include experiences that promote healing, understanding what led harm doers to their actions, having received help and having helped oneself or others at the time of one's suffering, caring by others, and prosocial role models. We suggest psychological changes that may result from these influences and lead to altruistic action: strengthening of the self, a more positive orientation toward people, empathy and belief in one's personal responsibility for others' welfare. The article critically reviews relevant research, and suggests future research directions and interventions to promote altruism born of suffering. Given the amount of violence between individuals and groups, understanding how victims become caring rather than aggressive is important for promoting a more peaceful world. PMID- 19123747 TI - Is there intergenerational transmission of trauma? The case of combat veterans' children. AB - This article is a review of the literature on intergenerational transmission of posttraumatic stress disorder (PTSD) from fathers to sons in families of war veterans. The review addresses several questions: (1) Which fathers have a greater tendency to transmit their distress to their offspring? (2) What is transmitted from father to child? (3) How is the distress transmitted and through which mechanisms? And finally, (4) Which children are more vulnerable to the transmission of PTSD distress in the family? Whereas the existing literature deals mainly with fathers' PTSD as a risk for increased emotional and behavior problems among the children, this review also highlights the current paucity of knowledge regarding family members and extrafamilial systems that may contribute to intergenerational transmission of PTSD or to its moderation. Little is also known about resilience and strengths that may mitigate or prevent the risk of intergenerational transmission of trauma. PMID- 19123748 TI - Development and validation of the child psychosocial distress screener in Burundi. AB - In non-Western countries, efficient and contextually valid methods of community screening are scarce. The present study describes the validation of a new, brief, 7-item multi-informant screener for conflict-affected children (Child Psychosocial Distress Screener; CPDS). To determine concurrent validity, the CPSD was administered to 65 children and their teachers. CPDS scores were compared with indication for psychosocial treatment based on an in-depth clinical assessment by a psychiatrist and psychologist. Construct validity was assessed by testing the measurement equivalence of the CPDS in a community sample (N = 2,240) in Burundi. The CPDS identifies indication for treatment with an accurateness of .81(sensitivity of .84; specificity of .60). Test?retest reliability of the instrument is good (.83). A robust and invariant factor structure provides evidence for the construct validity of the CPSD. The CPDS appears to be a useful multidimensional tool that measures nonspecific child psychosocial distress, detecting children with an indication for treatment. Because of brevity and the ability to be administered by nonspecialists, the CPDS can be an appropriate instrument to screen large populations of conflict-affected children. PMID- 19123749 TI - Pathways to posttraumatic growth versus posttraumatic stress: coping and emotional reactions following the September 11, 2001, terrorist attacks. AB - Exposure to trauma can lead to both posttraumatic growth and posttraumatic stress, but little is known about the commonalities and differences in the pathways through which they occur. The authors examined coping and emotional reactions as mediators of the effect of television exposure on both posttraumatic growth and posttraumatic stress in a nationally representative sample of 1,004 U.S. adults approximately 6 weeks after the September 11, 2001, terrorist attacks. Although posttraumatic growth and posttraumatic stress symptoms were moderately positively related, the pathways from coping and emotions to the outcomes differed: Positive coping and anger were more strongly related to posttraumatic growth than to posttraumatic stress, and pathways of negative coping and feeling depressed regarding the attacks were more strongly related to stress than to growth. Comparison of models suggested that emotions are both outcomes of and motivators for coping and that patterns of coping and emotions relate differentially to posttraumatic stress and posttraumatic growth. PMID- 19123751 TI - Attachment-based intervention for maltreating families. AB - This article presents attachment theory-based intervention strategies as a means of addressing the core parent-child interaction deficits that characterize homes in which children are exposed to maltreatment. The article outlines the socioemotional and cognitive outcomes of maltreatment and proposes that although many prevention programs target different parental and family characteristics, few address the core relationship issues that are at stake. Recent research on attachment-based intervention strategies, aimed at improving the sensitivity and responsiveness of the parenting behaviors that children are exposed to, are presented as providing a means of addressing this domain. Attachment theory and research are briefly summarized, and the relational and interactional patterns observed in maltreating families, and their link to infant and child developmental outcome, are described. Research on attachment-based intervention is addressed, with a focus on studies conducted in the context of maltreating or high-risk families. This work is synthesized to present the basic components viewed as critical to effective attachment intervention with maltreating families. Finally, the authors end with recommendations aimed at the effective implementation of attachment-based intervention. PMID- 19123750 TI - Relations among gender, violence exposure, and mental health: the national survey of adolescents. AB - Using a nationally representative sample of 4,008 adolescents, this study examines gender differences in violence exposure, major depressive episode (MDE) and posttraumatic stress disorder (PTSD), and characteristics of violence incidents. It was hypothesized that there would be gender differences in the types of violence exposure reported and in the prevalence of MDE and PTSD and that gender would moderate the relationship between violence exposure and mental health outcomes. Results indicated significant gender differences in rates of violence exposure, PTSD, and MDE. Additionally, gender was a moderating variable in the relation between sexual assault and PTSD, but not in the other violence exposure?mental health relations examined. It thus appears that the pathways for developing PTSD may be different for male and female victims of sexual abuse. Implications for interventions and future research are discussed. PMID- 19123752 TI - Social relationships among persons who have experienced serious mental illness, substance abuse, and homelessness: Implications for recovery. AB - The new paradigm of recovery has highlighted the importance of positive social relationships, but little is known about their role in recovery among homeless individuals with serious mental illness and comorbid substance abuse. This study used within- and across-case analyses of longitudinal data from qualitative interviews with 41 dually diagnosed individuals entering residential programs to exit homelessness and receive needed services. Thematic findings include (a) "loner talk" and the need for privacy; (b) family ties as "good news, bad news"; (c) when it comes to a partner, other things come first; and (d) in search of positive people. Analyses of change in individual trajectories revealed that stronger social relationships did not coincide exactly with positive outcomes. Although positive life changes were gradual, negative changes could be precipitous. Social relationships were threatened by concentrated disadvantage, that is, a lack of social and economic currency. Findings are discussed with implications for improving services for the most vulnerable individuals who stand to benefit from the era of recovery. PMID- 19123753 TI - Substance involvement among youths in child welfare: the role of common and unique risk factors. AB - This study examines risk factors for substance involvement for youths involved with the child welfare (CW) system. In addition to common risk factors examined in general population studies; this research examines risk factors unique to youths in the CW system, including age at entry into CW and number of out-of-home placements. Participants included 214 youths ages 13 to 18, randomly sampled from youths active to CW in San Diego County, California. Severity of substance involvement was assessed through structured diagnostic interviews determining lifetime substance use, abuse, and dependence. Hierarchical regression analyses including demographics, psychosocial variables, maltreatment history, CW placement variables, and the interaction of age at entry into CW and number of out-of-home placements revealed that both common and CW-specific risk factors were associated with the severity of youth substance involvement. Multiple placement changes, later entry into the CW system, and multiple-placement changes at an older age are associated with higher risk for more serious substance involvement for youths in CW. PMID- 19123754 TI - On failed intersubjectivity: Recollections of loneliness experiences in offspring of Holocaust survivors. AB - Intergenerational consequences of extensive trauma experienced by parents for the loneliness experienced by their children were explored in 52 adults (26 men and 26 women) who grew up in Holocaust survivor families. These adults, children of mothers who had survived Nazi concentration camps, were recruited from a random nonclinical Israeli sample. A narrative analysis of their recollected accounts of loneliness in childhood and adolescence yielded 4 major categories of loneliness experiences in the context of growing up in Holocaust survivor families: (a) echoes of parental intrusive traumatic memories; (b) echoes of parental numbing and detachment; (c) perceived parents' caregiving style; and (d) social comparison with other families, in particular the lack of grandparents. The echoes of the parental trauma in the recollected loneliness accounts are conceptualized as representing a sense of failed intersubjectivity in these interpersonal processes. The experiences of not being understood by others, not understanding others, and the lack of shared understanding involved in failed intersubjectivity are discussed and related to the importance of opening lines of communication between survivors and their descendents. PMID- 19123755 TI - Attitudes toward and understanding of children's rights among middle school students in Jerusalem: the role of family values and patterns, nationality, and religion. AB - Children's rights have become a cornerstone of discussions of human rights and human services around the world. However, the meaning of children's rights and their significance for policies and programs vary across nations, cultures, religions, and families. Only recently has research begun to study the conceptualization of children's rights in non-Western and non-Christian-dominated cultures and, thus, in more traditional and authoritarian families. This article reports on a cross-sectional survey among 810 Jewish and 582 Arab children (372 Muslims, 210 Christians), aged 12-14 from eastern and western Jerusalem to examine how children view their rights. Adolescents completed a structured, anonymous, self-report questionnaire. Results suggest that nationality/ethnicity is a major factor in explaining differences in children's views of their rights, with religion playing a minor role. Results also suggest that family values and practices are significantly correlated with the approach to children's rights, as is gender, although its explanatory power is weaker. These findings should guide practitioners and children's rights advocates as they strive to enhance the support for children's rights in the Middle East and develop appropriate policies. PMID- 19123756 TI - Jewish and non-Jewish World War II child and adolescent survivors at 60 years after war: effects of parental loss and age at exposure on well-being. AB - The study investigated the effects of World War II (WWII) on psychological and social functioning of Jewish and non-Jewish survivors 60 years after the war. The authors hypothesized that the level of posttraumatic symptoms, depression, and social isolation of survivors who were at least 5 years old (but younger than 18) in the last year of WWII would be predicted by the extent of traumatic loss, (i.e., death of parent[s]) and age at the end of WWII. Data were collected from 211 individuals living in Poland, ages 66-80; 30% were Jewish Holocaust survivors. Current posttraumatic stress disorder was almost 2 times higher for Jewish (55.6%) than for non-Jewish survivors (30.9%), whereas no differences were found for depression and social isolation. Parental loss during the war predicted a global decrement of well-being (across measured outcome indices). For certain subgroups (e.g., Jewish survivors who had not lost their parents during WWII), war trauma may have less profound effects if most of the trauma exposure occurred during an earlier age (i.e., <5 years). PMID- 19123757 TI - Development of the zoological garden and mental hospital. AB - Ellenberger in 1960 described the parallel histories of the zoological garden and the mental hospital. To determine how impressions of the 2 institutions have evolved in the intervening years, students in an abnormal psychology class rated several mental hospital and zoo terms. The zoo was rated more positively than the mental hospital, but not as favorably as the wild animal park or the nature reserve. These ratings reflect the changes in zoo design and management that have occurred in the intervening years. Improvements in mental hospital design and mission were largely derailed by the deinstitutionalization movement. A second study, intended to examine the fruitfulness of Ellenberger's comparison, asked students about the "show value" of patients in different diagnostic categories. There was a negative relationship between preference for seeing patients on a tour, in which the major psychoses were top choices, and clients with less severe disorders with whom students would prefer to work professionally. PMID- 19123758 TI - Call to action: Reducing interpersonal violence across the lifespan. AB - On September 12-13, 2008, more than 40 representatives from national professional organizations, grass roots organizations, governmental agencies, policy organizations, and advocacy movements gathered together in San Diego, California, as part of the Think Tank on Reducing Interpersonal Violence Across the Lifespan: Connecting Agendas. Working as a whole and in small groups, the think tank identified major gaps and issues facing the field of interpersonal violence prevention and began to develop a blueprint for integrating action across types of violence and types of populations, as well as integrating research, practice, and policy across the lifespan. Recognizing the need for a mechanism to facilitate communication and integrative collaboration in research, practice, and policy activities across diverse domains of interests and populations, the organizations committed to work together formed the National Partnership to Prevent Interpersonal Violence Across the Lifespan. The 2010 conference will bring together researchers, practitioners, policymakers, advocates, consumers, and funders in a series of plenary sessions, workshops, and poster sessions to present cutting-edge examples of integrative work aimed at transforming approaches to the prevention of interpersonal violence. Interpersonal violence is an intransigent problem with widespread and long-lasting direct and indirect effects. Rates of intimate partner violence (IPV), sexual assault, and childhood physical and sexual abuse around the world remain unacceptably high (CDC, 2006; see also http://www.cdc.gov/ncipc). Although the rippling effects of interpersonal violence have yet to be fully understood and documented, it is known that outcomes of violence encompass death, injury, and a wide range of physical, sexual, reproductive, and mental health problems. This special section of the American Journal of Orthopsychiatry encourages support for the call to action by the National Partnership to Prevent Interpersonal Violence Across the Lifespan. PMID- 19123759 TI - National and global agendas on violence against women: historical perspective and consensus. AB - A policy analysis of 11 national and global institutions' violence against women agendas spanning 1990 to 2006 is presented. Analysis revealed 85 distinct recommendations. The highest percentages of them referenced prevention (29%); data, design, and measurement (21%); and psychotherapy and support (19%). Consensus (percentage of recommendations for future activities included in four or more agendas) was highest for advocacy (75%), funding (50%), prevention (48%), and data, design, and measurement (44%). Changes in emphasis over time, aims that have been abandoned, and observations contrasting U.S. and global agendas are also examined. The results create a context to inform the agendas currently in development within psychology, criminal justice, medicine, nursing, public health, and other disciplines. Next steps to guide future policy work include investigation of advocates', practitioners', researchers', and policymakers' perceived progress in implementing existing recommendations, empirical cataloguing of achievements that demonstrate progress toward aims, constituent input on reprioritization of activities, and contemporizing action steps. PMID- 19123760 TI - The complexity of trauma types in the lives of women in families referred for family violence: Multiple mediators of mental health. AB - Responding to calls for further research about the impact of multiple types of trauma across the life span, this study examines the interconnections among types of trauma in childhood and adulthood in a convenience clinical sample of 283 women obtaining social services for family violence. In particular, variables including family of-origin dysfunction and other childhood risk factors, relationship victimization in adulthood, and the presence of adult resources were examined as mediators of links between child maltreatment and adult mental health symptoms. Variables were assessed at different time points, 3 years apart. Path analysis revealed that the conceptual model of multiple pathways between childhood family violence exposure and adult outcomes fit the data well. In particular, the link between child maltreatment and adult trauma symptoms was mediated by more proximal adult sexual and intimate partner violence and its association with childhood risk markers (e.g., negative family environment) and decreased markers of resources. This link was not significant for a more general index of mental health symptoms in adulthood. PMID- 19123761 TI - Examining the sexual offenses of female juveniles: The relevance of childhood maltreatment. AB - Research on female juvenile sex offenders is limited by small clinical samples. Little is known about the characteristics of female sexual offending and how it is related to child maltreatment. This study examines data from the case histories of 118 female juvenile sex offenders. In contrast to portrayals in previous research, this study shows that female sex offenders are not a homogeneous group. Findings also included differentiation between female juvenile sexual offenders with a history of child maltreatment and those without a history of child maltreatment. Female juvenile sex offenders who had a history of child maltreatment were more likely to have a current mental health diagnosis and experience clinical levels of anger--irritability and depression--anxiety than those without a history of child maltreatment. The impact of a history of sexual abuse for female juvenile sex offenders was found to be important with regard to higher levels of coercion of their sexual abuse victims. Important distinctions are highlighted that have implications for female-specific assessment, treatment, and prevention. PMID- 19123762 TI - The role of socioeconomic status in helpseeking from hotlines, shelters, and police among a national sample of women experiencing intimate partner violence. AB - The role of socioeconomic status (SES) in the prevalence and course of intimate partner violence (IPV) has been established, but mechanisms of this role are less clear. An untested assumption has been that women of greater resources are better able to seek help. This study used a national sample of 1,077 women who had experienced IPV to explore the role of income and education in helpseeking from hotlines, shelters, and police. The authors found that SES did not play a large role in the use of hotlines, the least often used service in this study. Women with more income were less likely to use shelters and were less likely to feel they should have used them. There was an interaction between income and severity of violence in predicting police use, such that severity of violence only predicted calling police among higher income women. This finding suggests the possibility of a lower threshold for reporting violence among the lowest income women. The authors discuss the need for research using a more diverse sample of women, and the need to fine tune services according to our emerging understanding of social context. PMID- 19123763 TI - Acute stress responses: A review and synthesis of ASD, ASR, and CSR. AB - Toward the development of a unifying diagnosis for acute stress responses this article attempts to find a place for combat stress reaction (CSR) within the spectrum of other defined acute stress responses. This article critically compares the diagnostic criteria of acute stress disorder (ASD), acute stress reaction (ASR), and CSR. Prospective studies concerning the predictive value of ASD, ASR, and CSR are reviewed. Questions, recommendations, and implications for clinical practice are raised concerning the completeness of the current acute stress response diagnoses, the heterogeneity of different stressors, the scope of expected outcomes, and the importance of decline in function as an indicator of future psychological, psychiatric, and somatic distress. PMID- 19123764 TI - Psychiatric symptom patterns, proximal risk factors, and sexual risk behaviors among youth in outpatient substance abuse treatment. AB - The purpose of the current study was to classify adolescents receiving outpatient treatment for alcohol or other drug (AOD) problems via self-reports of the Diagnostic and Statistical Manual of Mental Disorders (4th ed. American Psychiatric Association, 1994) psychiatric symptoms, and to identify group differences in measures of proximal risk factors for sexual risk behaviors (SRBs) and self-reported SRBs. Structured interviews were administered to 300 adolescent clients (202 males, 98 females; M = 16.22 years; SD = 1.13) receiving treatment services as part of a larger National Institute on Alcohol Abuse and Alcoholism (NIAAA)-funded randomized clinical trial. Ward's method cluster analysis (Ward, 1963) was used to classify adolescents into distinct groups based on psychiatric symptom profiles. A multivariate analysis of variance was used to identify significant between-cluster differences in self-reported SRBs and proximal risk factors for SRB. Substantial heterogeneity in patterns of psychiatric symptoms was documented in this treatment sample. Membership in certain psychiatric symptom clusters was associated with several self-reported SRBs and correlated proximal risk factors. Among youth receiving AOD treatment, interventions to promote HIV/STI risk reduction may need adaptation for those with differing psychiatric profiles. PMID- 19123765 TI - Deliberate self-harm and alcohol involvement in college-aged females: a controlled comparison in a nonclinical sample. AB - Individuals who engage in deliberate self-harm (DSH) report using other problematic coping mechanisms. One potential problematic coping mechanism is alcohol consumption. Research on alcohol involvement and deliberate self-harm is conflicting. This study compared individuals who have engaged in deliberate self harm to controls on a range of alcohol measures. Five hundred females completed questionnaires assessing deliberate self-harm and alcohol involvement. Controlling for differences in psychopathology and impulsivity, the DSH group did not differ from the controls relative to quantity and frequency of alcohol use but did differ relative to negative consequences, risky behaviors, and alcohol expectancies. The authors discuss mechanisms that account for increased negative consequences and expectancies of alcohol consumption in the absence of differences in quantity and frequency, as well as the clinical importance of assessing a broad spectrum of alcohol involvement in the DSH population. PMID- 19123766 TI - Tics, twitches, tales: The experiences of Gilles de la Tourette's syndrome. AB - Gilles de la Tourette's syndrome (GTS) is well described and although many clinicians are aware of the condition, misunderstandings about the disorder still persist. There have been few accounts of the experiences of people with GTS, and none from affected medical practitioners. The first personal account in the medical literature was over 25 years ago. This is the first personal account of having GTS written by a practicing physician. The sensation of having a tic is a peculiar one and is difficult to describe to those who have not experienced them for themselves. Tics are distinctly voluntary, though not wanted. The desire to tic stems from the relief of a physical sensation (that is distinct from a conventional somatic sensation) that builds up prior to the tic and is relieved by it. However, akin to scratching a mosquito bite, tics lead to more tics. The sensation leading to the tic is paradoxically relieved yet restimulated by the tic itself. The effect of GTS on a personal life story is recounted, and the condition briefly reviewed. PMID- 19123767 TI - Working with families in Assertive Community Treatment (ACT): the case manager's perspective. AB - In response to research findings of insufficient family involvement in mental health services for people with severe mental illness, this grounded theory study examines case managers' interactions with families of clients in Assertive Community Treatment (ACT). Findings suggest that case managers conceptualize families as sources of social connections, rather than sources of care, for clients. This conceptualization is influenced by case managers' goals, which also guide their assessments of families for involvement in treatment in terms of the extent to which families help attain treatment goals. In developing strategies to work with families, case managers engage in ongoing assessments and consider client permission for family involvement in treatment, family availability to clients, and family wishes for involvement in treatment. Three case examples illustrate the relationships among case managers' goals, assessments of families, and selections of work strategies. The potential role of the ACT model in shaping this particular view on families is also discussed. PMID- 19123768 TI - 'I used to dream of lupus as some sort of creature': Chronic illness as an internal object. AB - This study examines the place occupied by chronic illness in the inner lives of 15 women suffering from Systemic Lupus Erythematosus (SLE). A phenomenological analysis of illness narratives demonstrates that sufferers construe their illness as a protagonist or, using an object-relations informed perspective, as an internal object. That is, with time sufferers constituted a mental representation of SLE that in itself has the power to influence the sufferers' affective states and behaviors. An insight into these "illness relations" is conducive to a better understanding of the "lived experience" of SLE for disabled, economically disadvantaged women. Through their experience, the study of risk and resilience in chronic illness may be advanced. PMID- 19123769 TI - Identity development and psychological well-being in Korean-born adoptees in the U.S. AB - Because adult Korean-born adoptees have unique experiences, the factors that contribute to their psychological well-being need to be studied separately from both Caucasian and Korean Americans. In this Internet-based study with 83 adult Korean-born adoptees in the United States, both ethnic identity and adjustment to adoption (considered a component of adoptive identity) were expected to predict psychological well-being. Results supported predictions: Each measure of psychological well-being (personal growth, self-acceptance, and positive relationships with others) was affected by the predictive variables in unique ways. Cultural socialization experiences also were related to personal growth, but this association was fully mediated by strength of ethnic identity. Implications for adoptive parents and counselors are discussed. PMID- 19123770 TI - How expressions of forgiveness, purpose, and religiosity relate to emotional intelligence and self-concept in urban fifth-grade students. AB - This study investigated how the tendency to express forgiveness, purpose, and religiosity in a free-response essay relates to emotional intelligence and self concept in 89 5th-graders (mean age = 10.84 years) from an urban public school district in New Jersey. Readers coded essays for expressions of forgiveness, purpose, and religiosity using originally developed rubrics. These data were compared with self-reports on scales of emotional intelligence and self-concept. It was hypothesized that expressions of the predictor variables would correlate positively with emotional intelligence and self-concept. In contrast to expressions of purpose, which were common among students, expressions of forgiveness and religiosity were infrequent. Furthermore, forgiveness was not significantly related to either criterion variable; purpose was positively related to self-concept (but not to emotional intelligence); and religiosity was negatively related to emotional intelligence (but not to self-concept). Correlational analyses by gender revealed a possible trend toward more robust relationships being observed among females than males; however, the differences between the correlation coefficients observed among males and females failed to reach statistical significance. Several of the study's unanticipated findings suggest the need for further empirical work investigating the psychological correlates of these constructs in children. PMID- 19123771 TI - The association between timely outpatient visits and the likelihood of rehospitalization for schizophrenia patients. AB - Early rehospitalization is common in schizophrenia. Given the impact on cost containment, there is an urgent need to identify the predictors of rehospitalization. This study aims to examine the association between the frequency of timely outpatient visits and the likelihood of early rehospitalization in schizophrenia patients. After adjusting for gender, age, substance use, and length of stay, patients failing to attend any outpatient appointments within 2 months after discharge had significantly higher rehospitalization rates than those attending at least 1 appointment. Thus, timely outpatient visits, shortly after discharge, may be associated with a decreased risk of rehospitalizations. PMID- 19123772 TI - Relationship of stigma to HIV risk among women with mental illness. AB - Urban women with severe mental illness (SMI) are vulnerable to stigma and discrimination related to mental illness and other stigmatized labels. Stigma experiences may increase their risk for negative health outcomes, such as HIV infection. This study tests the relationship between perceived stigma and HIV risk behaviors among women with SMI. The authors interviewed 92 women attending community mental health programs using the Stigma of Psychiatric Illness and Sexuality Among Women Questionnaire. There were significant relationships between personal experiences of mental illness and substance use accompanying sexual intercourse; perceived ethnic stigma and having a riskier partner type; and experiences of discrimination and having a casual or sex-exchange partner. Higher scores on relationship stigma were associated with a greater number of sexual risk behaviors. The findings underscore the importance of exploring how stigma attached to mental illness intersects with other stigmatized labels to produce unique configurations of HIV risk. HIV risk reduction interventions and prevention research should integrate attention to stigmatized identities in the lives of women with SMI. PMID- 19123773 TI - Multimode gradient elution in reversed-phase liquid chromatography: application to retention prediction and separation optimization of a set of amino acids in gradient runs involving simultaneous variations of mobile-phase composition, flow rate, and temperature. AB - The theory of multimode gradient elution in liquid chromatography involving combined gradients of the mobile-phase composition with flow rate and column temperature is presented, and a very simple stepwise method that allows for the calculation of the elution time of a sample solute under all gradient conditions is proposed. The theory is successfully applied to the separation of 12 o phthalaldehyde derivatives of amino acids in eluting systems modified by acetonitrile. Average errors below 2.9% have been found in the retention prediction using the above method, which is supported by adequate models and algorithms capable of describing the chromatographic behavior of solutes upon changes in the separation factors, such as the modifier content, flow rate, and temperature. PMID- 19123774 TI - User configurable microfluidic device for multiplexed immunoassays based on DNA directed assembly. AB - We present a microfluidic device for multiplexed immunoassays based on DNA directed immobilization. Because of the versatile building blocks used for this technique, it is possible to build up user configurable protein microarrays within the microfluidic system by means of DNA-directed self-assembly, which can be used for immunoassay applications. We demonstrate the performance of our system by parallel detection of cytokines in a multiplex immunoassay, employing silver deposition labeling and optical read-out. PMID- 19123775 TI - Stimuli-responsive polymersomes for programmed drug delivery. AB - In the past decade, polymersomes (also referred to as polymeric vesicles) have attracted rapidly growing interest based on their intriguing aggregation phenomena, cell and virus-mimicking dimensions and functions, as well as tremendous potential applications in medicine, pharmacy, and biotechnology. Unlike liposomes self-assembled from low molecular weight lipids, polymersomes are in general prepared from macromolecular amphiphiles of various architectures including amphiphilic diblock, triblock, graft and dendritic copolymers. Polymersomes exhibit very unique features highlighted with high stability, tunable membrane properties, versatility, and capacity of transporting hydrophilic as well as hydrophobic species such as anticancer drugs, genes, proteins, and diagnostic probes. Recently, much effort has been directed to the development of intelligent polymersomes that respond to internal or external stimuli, in particular, pH, temperature, redox potential, light, magnetic field, and ultrasound, either reversibly or nonreversibly. Stimuli-sensitive polymersomes have emerged as novel programmable delivery systems in which the release of the encapsulated contents can be readily modulated by the stimulus. The stimuli-responsive release may result in significantly enhanced therapeutic efficacy and minimized possible side effects. It is also feasible to form and disassemble polymersomes in water simply by applying an appropriate stimulus. In this article, recent advances in stimuli-sensitive polymersomes have been reviewed, and perspectives on future developments have been discussed. PMID- 19123776 TI - Photoionization and vibrational spectroscopy of the aniline-methanol clusters. AB - Aniline-methanol mixed clusters are ionized by single photon vacuum ultraviolet (VUV, 118 nm) radiation with which absorption to an excited intermediate S(1) state is not required. Aniline ion (An(+)), a series of (An)(n)(+)-(CH(3)OH)(m) (n = 1, 2) cluster ions, and their hydrogenated cluster ions, (An)(n)(+) (CH(3)OH)(m)H (n = 1, 2) are observed by mass spectrometry. Infrared (IR) absorption spectra of aniline-methanol cluster cations and neutrals are measured through IR and VUV (118 nm) "ion dip" spectroscopy in the range 2500-4000 cm(-1). The observed mid-IR spectrum of the An(+)-CH(3)OH has two sharp absorption bands, at 3438 and 3668 cm(-1), which are assigned to the free NH stretch vibration of the aniline cation and the free OH stretch vibration of methanol, respectively. Calculations demonstrated that a change in the charges on the nitrogen atom of the amine group upon ionization of the neutral to the cluster cation alters the role of aniline from hydrogen acceptor to hydrogen donor in its interaction with methanol. Theoretical and experimental results suggest that a hydrogen bond forms between one of the H atoms of the aniline amine group and the lone pair of electrons of the methanol oxygen atom in the aniline-methanol cluster cation. Measured IR spectra and theoretical results for neutral clusters suggest that the H atom of the methanol OH moiety is bonded to the aniline amine group lone pair electrons for the neutral ground state aniline-methanol cluster. PMID- 19123777 TI - Role of directed van der Waals bonded interactions in the determination of the structures of molecular arsenate solids. AB - Bond paths, local energy density properties, and Laplacian, L(r) = wedge(2)rho(r), composite isosurfaces of the electron density distributions were calculated for the intramolecular and intermolecular bonded interactions for molecular solids of As(2)O(3) and AsO(2) composition, an As(2)O(5) crystal, a number of arsenate molecules, and the arsenic metalloid, arsenolamprite. The directed intermolecular van der Waals As-O, O-O, and As-As bonded interactions are believed to serve as mainstays between the individual molecules in each of the molecular solids. As-O bond paths between the bonded atoms connect Lewis base charge concentrations and Lewis acid charge depletion domains, whereas the O-O and As-As paths connect Lewis base pair and Lewis acid pair domains, respectively, giving rise to sets of intermolecular directed bond paths. The alignment of the directed bond paths results in the periodic structures adopted by the arsenates. The arrangements of the As atoms in the claudetite polymorphs of As(2)O(3) and the As atoms in arsenolamprite are similar. Like the As(2)O(3) polymorphs, arsenolamprite is a molecular solid connected by relatively weak As As intermolecular directed van der Waals bond paths between the layers of stronger As-As intramolecular bonded interactions. The bond critical point and local energy density properties of the intermolecular As-As bonded interactions in arsenolamprite are comparable with the As-As interactions in claudetite I. As such, the structure of claudetite I can be viewed as a stuffed derivative of the arsenolamprite structure with O atoms between pairs of As atoms comprising the layers of the structure. The cubic structure adopted by the arsenolite polymorph can be understood in terms of sets of directed acid-base As-O and base-base O-O pair domains and bond paths that radiate from the tetrahedral faces of its constituent molecules, serving as face-to-face key-lock mainstays in forming a periodic tetrahedral array of molecules rather than one based on some variant of close packing. The relatively dense structure and the corrugation of the layers in claudetite I can also be understood in terms of directed van der Waals As-O bonded interactions. Our study not only provides a new basis for understanding the crystal chemistry and the structures of the arsenates, but it also calls for a reappraisal of the concept of van der Waals bonded interactions, how the structures of molecular solids are viewed, and how the molecules in these solids are bonded in a periodic structure. PMID- 19123778 TI - Structural insight into G-protein coupled receptor binding by apelin. AB - Apelin peptides are the cognate ligands for the G-protein coupled receptor APJ, with functions in the cardiovascular and central nervous systems, in glucose metabolism and as a human immunodeficiency virus (HIV-1) coreceptor. Apelin is found in 13-36 residue forms in vivo. The structures of five isoforms of apelin at physiological versus low (5-6 degrees C) temperature are compared here using circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy, demonstrating increased structure at low temperature. Far-ultraviolet (UV) CD spectra are predominantly random coil for apelin isoforms, but are convoluted by unusual bands from the C-terminal phenylalanine side chain. These bands, assigned using F13A-apelin-13, are accentuated at 5 degrees C and imply conformational restriction. At 35 degrees C, the R6-L9 region of apelin-17 is well structured, consistent with previous mutagenesis results showing necessity of this segment for apelin-APJ binding and activation. At 5 degrees C, R6-L9 retains its structuring while the functionally critical C-terminal G13-F17 region also becomes highly structured. Type IV beta-turns and some polyproline-II structure alongside F17 side chain motional reduction correlate well with CD spectral properties. Cis-trans peptide bond isomerization at P14 and P16 produces two sequentially assignable conformers (both trans:both cis approximately 4:1) alongside less populated conformers. Chemical shift assignment of apelin-12, -13 and pyroglutamate-apelin-13 implies highly similar structuring and the same isomerization at the C-terminus. Based on the apelin-17 structure, a two-step binding and activation mechanism is hypothesized. PMID- 19123779 TI - A self-starting method for obtaining analytic potential-energy surfaces from ab initio electronic structure calculations. AB - Previous methods proposed for obtaining analytic potential-energy surfaces (PES) from ab initio electronic structure calculations are not self-starting. They generally require that the sampling of configuration space important in the reaction dynamics of the process being investigated be initiated by using chemical intuition or a previously developed semiempirical potential-energy surface. When the system under investigation contains four or more atoms undergoing three- and four-center reactions in addition to bond scission processes, obtaining a sufficiently converged initial sampling can be very difficult due to the extremely large volume of configuration space that is important in the reaction dynamics. It is shown that by combining direct dynamics (DD) with previously reported molecular dynamics (MD), novelty sampling (NS), and neural network (NN) methods, an analytical surface suitable for MD computations for large systems may be obtained. Application of the method to the investigation of N-O bond scission and cis-trans isomerization reactions of HONO followed by comparison of the resulting neural network potential-energy surface to one obtained by using a semiempirical potential to initiate the sampling shows that the two potential surfaces are the same within the fitting accuracy of the surfaces. It is concluded that the combination of direct dynamics, molecular dynamics, novelty sampling, and neural network fitting provides a self-starting, robust, and accurate DD/MD/NS/NN method for the execution of first-principles, ab initio, molecular dynamics studies in systems containing four or more atoms which are undergoing simultaneous two-, three-, and four-center reactions. PMID- 19123780 TI - Dimeric complexes of tryptophan with M2+ metal ions. AB - IRMPD spectroscopy using the FELIX free electron laser and a Fourier transform ICR mass spectrometer was used to characterize the structures of electrosprayed dimer complexes M(2+)Trp(2) of tryptophan with a series of eight doubly charged metal ions, including alkaline earths Ca, Sr, and Ba, and transition metals Zn, Cd, Mn, Co, and Ni. With the support of DFT thermochemical calculations, at least three different structural motifs were distinguished spectroscopically, depending critically on the nature of the metal ion. The spectral signatures of a ligand in the charge-solvated (CS) configuration, namely peaks near 1730 and 1150 cm(-1), were prominent in all the spectra, and it was clear that all the dimer complexes contain at least one CS ligand. The spectra indicated that the second ligand is zwitterionic (ZW) for all complexes except the Ni case, with the second ligand having an extended binding geometry with smaller metals but showing some admixture of a compact chelated geometry with larger alkaline earths. It was concluded that these dimer complexes have a mixed configuration of ligands, denoted CS/ZW. The Ni(2+)Trp(2) complex is exceptional, with the spectroscopy and the thermochemical calculation both indicating a CS/CS configuration of ligands. This geometry appears to correlate with the exceptionally small size and high binding strength of the Ni(2+) cation. The complex CdClTrp(1+) was also obtained and gave a clear spectrum showing a CS ligand configuration. The presence of a CS ligand in all the dimeric complexes of the 2+ metals is an interesting contrast with the monomer complex Ba(2+)Trp, in which the ligand is ZW. PMID- 19123781 TI - Microwave-assisted combinatorial synthesis of new 3-pyrimidin-5-ylpropanamides via a solvent-dependent chemoselective reaction. AB - A series of new 3-pyrimidin-5-ylpropanamides was selectively synthesized via a microwave-assisted, chemoselective reaction of arylidene-Meldrum's acid, 6 hydroxypyrimidin-4(3H)-one, and structurally diverse amines including (S or R)-1 phenylethanamine, cyclohexanamine, and cyclopentanamine depended on nature of solvents. In this reaction, the utilization of HOAc as a solvent leads to 3 pyrimidin-5-ylpropanamides, whereas water as reaction media results in the spiro[5.5]undecane-1,5,9-triones from same starting materials. This method has the advantages of short synthetic route, operational simplicity, increased safety for small-scale high-speed synthesis, and minimal environment impact. PMID- 19123782 TI - Stabilization of square planar silicon: a new building block for conjugated Si containing systems. AB - The feasibility of square planar silicon as a building block for conjugated systems was investigated by ab initio calculations. A five-membered ring model system was used to map electronic and steric substituent effects that might help in the stabilization of the planar structure. A pi push-pull arrangement around the silicon was found to prefer planarity. Aromaticity was proven to play an important role in stabilization as well. With the help of steric constraints, a new structure was proposed as a synthetic target containing square planar silicon. The kinetic stability of this structure was also investigated. PMID- 19123783 TI - Structures of copper complexes of the hybrid [SNS] ligand of bis(2 (benzylthio)ethyl)amine and facile catalytic formation of 1-benzyl-4-phenyl-1H 1,2,3-triazole through click reaction. AB - A hybrid ligand, bis(2-(benzylthio)ethyl)amine (SNS), with an amine and two thioether donors reacts with Cu(II) to give mononuclear [CuCl(2)(SNS)] (1), [CuBr(2)(SNS)] (2), [Cu(OTf)(2)(SNS)(OH(2))] (3), and an one-dimensional Cu(I) coordination polymer [Cu(2)I(2)(SNS)](n) (4). All complexes have been characterized by single-crystal X-ray diffraction analysis, and 1-3 were studied by EPR analysis at room temperature. Complexes 1 and 2 are penta-coordinated with a distorted square pyramidal metal supported by a tridentate SNS ligand on the basal plane. Complex 3 shows a tetragonally distorted octahedral sphere with two trans and weakly bonding monodentate triflates. A 12-membered ring in the solid lattice is formed by intermolecular H-bonding among the coordinated triflate and aqua ligands from four neighboring molecules. Complex 4, the only Cu(I) in this series, shows a coordination polymer chain [Cu(4)I(4)](n) comprising tetrahedral Cu(I) centers stitched by the SNS ligand in a unique bridge-chelate mode in the form of a helix. All four complexes are catalytically active at room temperature in a copper-catalyzed azide-alkyne cycloaddition (CuAAA) three-component click reaction of benzyl chloride, sodium azide, and phenylacetylene in an aqueous MeCN mixture to give good isolated yields of 1-benzyl-4-phenyl-1H-1,2,3-triazole, without the use of a base or reducing agent. PMID- 19123785 TI - Physico-chemical properties of iodine-adsorbed single-walled carbon nanotubes. AB - I2 was adsorbed on single-walled carbon nanotube from ethanol solution at 303 K. The I2 adsorption isotherm was Langmuirian, giving 35 (+/-10) mg g(-1) of the saturated adsorption amount (coverage 0.06-0.09). The I2-adsorption treatment of SWCNT bundles reduced the N2 adsorption amount at 77 K by only 3%; the adsorption amount of supercritical H2 at 77 K was decreased by 30% because of the I2 adsorption treatment, indicating the blocking of interstitial pores by adsorbed I2. These adsorption results indicated the adsorption of I2 molecules in the narrow interstitial pores. The I2-adsorption treatment increases the Raman intensity coming from metallic SWCNTs, and the dc electrical conductivity increased by 15% because of the I2-adsorption treatment, strongly suggesting the presence of charge-transfer interaction between I2 and SWCNTs irrespective of small coverage by I2. PMID- 19123786 TI - Boundary effect on diffusiophoresis: spherical particle in a spherical cavity. AB - The boundary effect on the diffusiophoretic behavior of a particle is analyzed theoretically by considering the diffusiophoresis of a charged sphere under arbitrary surface potential and double-layer thickness at an arbitrary position in an uncharged spherical cavity. We show that the phenomenon under consideration is governed by double-layer relaxation, chemiosmotic/diffusioosmotic flow, and two types of competing double-layer polarization. The presence of the cavity has a profound influence on the diffusiophoretic behavior of the particle, especially when the surface potential is high. For instance, the scaled diffusiophoretic velocity of the particle has a local maximum as the position of the particle varies; it may have a local maximum and local minimum as the thickness of the double-layer varies. The significance of the effect of double-layer relaxation depends upon the level of surface potential and magnitude of the electric Peclet number. PMID- 19123787 TI - Orthogonal synthesis of isoindole and isoquinoline derivatives from organic azides. AB - Alpha-azido carbonyl compounds bearing a 2-alkenylaryl moiety at the alpha position are found to be promising precursors for synthesis of isoindole and isoquinoline derivatives via 1,3-dipolar cycloaddition of azides onto alkenes and 6pi-electrocyclization of N-H imine intermediates, respectively. PMID- 19123788 TI - Identification of human pituitary growth hormone variants by mass spectrometry. AB - The heterogeneity of human endogenous growth hormone (GH) is used in doping control analysis to distinguish it from the homogeneous recombinant analogue in plasma samples. Pituitary GH variants were characterized by gel electrophoresis and mass spectrometry. Besides 22 and 20 kDa isoforms, fragments of 9 and 12 kDa were identified and a glycosylated 23 kDa GH variant was elucidated to bear a HexHexNac 2 NeuAc modification presumably located at Thr 60. PMID- 19123789 TI - Enzymatic formation of unnatural novel chalcone, stilbene, and benzophenone scaffolds by plant type III polyketide synthase. AB - A C(19) hexaketide stilbene and a C(21) heptaketide chalcone were synthesized by Aloe arborescens octaketide synthase (OKS), a plant-specific type III polyketide synthase (PKS). Remarkably, the C(21) chalcone-forming activity was dramatically increased in a structure-guided OKS N222G mutant that produces a C(20) decaketide SEK15 from 10 molecules of malonyl-CoA. The findings suggested further strategies for production of unnatural polyketides by combination of the precursor-directed biosynthesis and the structure-guided engineering of type III PKS. PMID- 19123790 TI - Correlation between bonding geometry and band gap states at organic-inorganic interfaces: catechol on rutile TiO2(110). AB - Adsorbate-induced band gap states in semiconductors are of particular interest due to the potential of increased light absorption and photoreactivity. A combined theoretical and experimental (STM, photoemission) study of the molecular scale factors involved in the formation of gap states in TiO(2) is presented. Using the organic catechol on rutile TiO(2)(110) as a model system, it is found that the bonding geometry strongly affects the molecular electronic structure. At saturation catechol forms an ordered 4 x 1 overlayer. This structure is attributed to catechol adsorbed on rows of surface Ti atoms with the molecular plane tilted from the surface normal in an alternating fashion. In the computed lowest-energy structure, one of the two terminal OH groups at each catechol dissociates and the O binds to a surface Ti atom in a monodentate configuration, whereas the other OH group forms an H-bond to the next catechol neighbor. Through proton exchange with the surface, this structure can easily transform into one where both OH groups dissociate and the catechol is bound to two surface Ti in a bidentate configuration. Only bidendate catechol introduces states in the band gap of TiO(2). PMID- 19123791 TI - trans-Difluoro complexes of palladium(II). AB - Complexes trans-[(Py)(2)Pd(Ph)(F)] (1; Py = pyridine) and trans-[(t BuPy)(2)Pd(Ph)(F)] (2; t-BuPy = 4-tert-butylpyridine) have been prepared from the corresponding iodides and AgF. Thermal decomposition of 1 and 2 in anhydrous benzene at 80 degrees C did not result in C-F bond formation, but Pd black and Ph(2) were produced instead, along with novel difluorides trans-[(Py)(2)Pd(F)(2)] (3) and trans-[(t-BuPy)(2)Pd(F)(2)] (4). Both 3 and 4, the first trans-difluoro d(8) square complexes, were independently synthesized from the corresponding diiodides and AgF and fully characterized. Contrary to filled/filled d(pi)-p(pi) repulsion considerations, the Pd-F bond distances in 3 and 4 are unprecedentedly short, being only 1.947(4)-1.958(4) A and shorter than those in 1 and 2 by 0.12 0.13 A. The mechanism of formation of 3 and 4 and bonding in these complexes are discussed. PMID- 19123792 TI - Fragrant unsaturated aldehydes elicit activation of the Keap1/Nrf2 system leading to the upregulation of thioredoxin expression and protection against oxidative stress. AB - Thioredoxin, a key molecule in redox regulation, and many redox enzymes are regulated through the antioxidant responsive element (ARE). To search for antioxidative constituents, we screened extracts from vegetables and found that the extracts of Perilla frutescens and Artemisia princeps have potent thioredoxin inducing activities. By activity-guided purification of Perilla frutescens extracts, we identified perillaldehyde as a novel thioredoxin inducer. Fragrant unsaturated aldehydes, such as trans-cinnamaldehyde, safranal, 2,4-octadienal, citral, trans-2, cis-6-nonadienal, and trans-2-hexenal showed the ability to activate ARE. Perillaldehyde-induced activation through the ARE was suppressed by the overexpression of wild-type Keap1, whereas sulforaphane-induced activation seemed to be partially suppressed. Mutant Keap1 (R272A/K287A or C273A/C288A) did not suppress this activation. Pretreatment with perillaldehyde reduced the H(2)O(2)-induced cytotoxicity. Thus, we show that fragrant unsaturated aldehydes from edible plants are novel thioredoxin inducers and ARE activators and may be beneficial for protection against oxidative stress-induced cellular damage. These results also suggest that perillaldehyde activates the Nrf2-Keap1 system and that the lysine and arginine residues juxtaposed to the critical cysteine residues of Keap1 are required for signal sensing. PMID- 19123793 TI - Meloxicam and meloxicam-beta-cyclodextrin complex in model membranes: effects on the properties and enzymatic lipolysis of phospholipid monolayers in relation to anti-inflammatory activity. AB - Meloxicam, a nonsteroidal anti-inflammatory drug (NSAID), is known as a selective cyclooxygenase-2 inhibitor. Cyclooxygenase-2 is a membrane protein, functionally coupled to an interfacial enzyme, phospholipase A2. Consequently, it may be supposed that the interactions of NSAIDs with lipid membranes play a role in the anti-inflammatory process. In order to investigate the mechanism of this process, Langmuir films formed with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2 dilauroyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3 phosphoethanolamine, or 1,2-myristoyl-sn-glycero-3-phosphoethanolamine were exposed to meloxicam and its beta-cyclodextrin inclusion complex. The monolayers were studied by measuring surface pressure, electric surface potential, Brewster angle micrographs, polarization-modulation infrared reflection-absorption spectra, and phospholipase A2 activity; the inclusion complex was studied using molecular modeling. The results obtained show that the monolayers formed in the presence of meloxicam and its complex are expanded and more liquid-like compared to pure lipids. Both compounds modify hydration of the lipid polar heads, orientation of the molecules, morphology of the domains, and the rate of lipolysis catalyzed by phospholipase A2. The latter effect may be involved in the anti-inflammatory activity of meloxicam. Importantly, the effects observed with the meloxicam-beta-cyclodextrin complex are more pronounced compared to those of the free meloxicam. This observation may be relevant for developing new meloxicam preparations with increased bioavailability. PMID- 19123794 TI - Organometallic enantiomeric scaffolding. Sequential semipinacol/1,5-"Michael like" reactions as a strategic approach to bridgehead-quaternary center aza[3.3.1]bicyclics: application to the total synthesis of (-)-adaline. AB - A nontraditional approach to the enantiocontrolled construction of quaternary center-bearing heteroatom-bridged bicyclo[3.3.1]nonanes (homotropanes) is reported that is based on organometallic enantiomeric scaffolding. This strategy takes advantage of the unique reactivity profiles of TpMo(CO)(2)(5-oxo-eta(3) pyranyl) and TpMo(CO)(2)(5-oxo-eta(3)-pyridinyl) scaffolds, and features a molybdenum-mediated semipinacol/1,5-"Michael-like" reaction sequence to establish the quaternary center and synthesize the bridged bicyclic structure. An asymmetric total synthesis of (-)-adaline highlights this methodology. PMID- 19123795 TI - A yoctoliter-scale DNA reactor for small-molecule evolution. AB - The center of DNA three-way junctions, constituting a yoctoliter (10(-24) L) volume, is applied as an efficient reactor to create DNA-encoded libraries of chemical products. Amino acids and short peptides are linked to oligonucleotides via cleavable and noncleavable linkers. The oligonucleotide sequences contain two universal assembling domains at the center and a distal codon sequence specific for the attached building block. Stepwise self-assembly and chemical reactions of these conjugates in a combinatorial fashion create a library of pentapeptides in DNA three-way junctions in a single reaction vessel. We demonstrate the formation of an evenly distributed library of 100 peptides. Each library member contains a short synthetic peptide attached to a unique genetic code creating the necessary "genotype-phenotype" linkage essential to the process of in vitro molecular evolution. Selective enrichment of the [Leu]-enkephalin peptide from an original frequency of 1 in 10 million in a model library to a final frequency of 1.7% in only two rounds of affinity selection is described and demonstrates successful molecular evolution for a non-natural system. PMID- 19123796 TI - Predicting gas adsorption in complex microporous and mesoporous materials using a new density functional theory of finely discretized lattice fluids. AB - We introduce a nonlocal on-lattice version of density functional theory (DFT) that allows for efficient modeling of fluids in complex inhomogeneous materials. In its previous implementations, classical DFT has required fine discretization of the fluid density. As a result, in studies of gas adsorption it has been used only in idealized pore models with high symmetry. Our new lattice DFT dramatically reduces the computational demand required to model simple fluids and hence can be efficiently applied to complex materials with multiple directions of asymmetry. We apply our new lattice DFT to study nitrogen adsorption in a slit pore with open ends and directly obtain the correct desorption hysteresis. We also apply our DFT to predict hydrogen adsorption accurately in an atomistic model of a metal-organic framework. PMID- 19123797 TI - Separation of peptides with polyionic nanosponges for MALDI-MS analysis. AB - A polymer brush consisting of 70% poly(N-isopropylacrylamide) (PNIPAAM) and 30% polymethacrylic acid (PMAA) was synthesized from gold substrates with a "grafting from" AIBN-type free-radical initiator. Fractionation of two peptides, bradykinin and buccalin, was accomplished in less than 120 s by placing a 30 pM (pH approximately 6.2) droplet onto the polymer brush substrate. The eluant containing the anionic buccalin is pipetted away for MALDI analysis while the cationic bradykinin adsorbed to the swollen anionic brush and was subsequently released by adding a droplet of formic acid to the substrate. This caused the brush to collapse and release the bradykinin, much like squeezing a sponge; these nanosponge substrates exhibited very high loading capacity (>2.0 mg/mL) compared to plasma-polymer-modified MALDI substrates. Ellipsometric measurements showed that complementary peptides adsorb rapidly while those of the same charge do not, and MALDI-MS analysis of the two fractions showed separation of both peptides. The adsorption of bradykinin was monitored over time, and 85% of the peptide had been adsorbed to the nanosponge in 1 min from a 0.5 mg/mL aqueous solution. PMID- 19123800 TI - Chemical bonding and electronic and magnetic structure in LaOFeAs. AB - Periodic density functional calculations using hybrid exchange-correlation functionals predict that LaOFeAs is a strongly frustrated antiferromagnetic insulator with important covalence between Fe and As, with evident similarities with cuprates. PMID- 19123799 TI - Initial bacterial deposition on bare and zeolite-coated aluminum alloy and stainless steel. AB - In this study, the impact of zeolite thin film coatings on bacterial deposition and "biofouling" of surfaces has been investigated in an aqueous environment. The synthesis of two types of zeolite coatings, ZSM-5 coated on aluminum alloy and zeolite A coated on stainless steel, and the characterization of the coated and bare metal surfaces are described. The extent of cell deposition onto the bare and zeolite-coated aluminum alloy and stainless steel surfaces is investigated in a parallel plate flow chamber system under a laminar flow conditions. The initial rates of bacterial transfer to the various surfaces are compared by utilizing a marine bacterium, Halomonas pacifica g, under a range of ionic strength conditions. H. pacifica g deposited onto bare metal surfaces to a greater extent as compared with cells deposited onto the zeolite coatings. The surface properties found to have the most notable effect on attachment are the electrokinetic and hydrophobicity properties of the metal and zeolite-coated surfaces. These results suggest that a combination of two chemical mechanisms hydrophobic and electrostatic interactions-contribute to the antifouling nature of the zeolite surface. Additional observations on the relative role of the hydrodynamic and physical phenomena are also discussed. PMID- 19123801 TI - Mechanism of DNA strand exchange at liposome surfaces investigated using mismatched DNA. AB - DNA strand exchange is of great importance in vivo for genetic recombination and DNA repair. The detailed mechanism of strand exchange is not understood in full detail despite extensive studies. Simplistic model systems in which molecular parameters can be varied independently are therefore of interest to study. We chose the surface of a positively charged liposome as a scaffold, which we recently demonstrated to be able to catalyze the exchange of fully complementary DNA oligonucleotides. We here study how single base pair mismatches affect the rate of strand exchange on the liposome surface. Interestingly, the rate of the exchange does not simply follow the stability of the duplex in solution, as determined by melting temperatures, but also depends sensitively on the position of the mismatch. For duplexes with similar melting temperatures, the exchange is much faster for a mismatch close to the end than for a mismatch in the middle of the sequence. Our results suggest that the single strands are stabilized by the liposome surface; therefore, the duplex is fraying more and the DNA opens up in a zipperlike fashion on the surface, increasing the probability of strand exchange. We also show that the competition between greater stability (higher Tm in solution) and higher concentration is important for the final composition of the duplex when a large excess of single strands is added to a complementary double stranded DNA. Finally, the similar exchange rate constants for fully base-paired duplexes on the liposome surface when adding fully matched single strands or single strands with a mismatched base indicate that the rate is governed largely by separation of the initial duplex and not by the formation of the product duplex. PMID- 19123804 TI - Functionalization of poly(dimethylsiloxane) surfaces with maleic anhydride copolymer films. AB - Combining advantageous bulk properties of polymeric materials with surface selective chemical conversions is required in numerous advanced technologies. For that aim, we investigate strategies to graft maleic anhydride (MA) copolymer films onto poly(dimethylsiloxane) (PDMS) precoatings. Amino groups allowing the covalent attachment of the MA copolymer films to the PDMS (Sylgard 184) surface were introduced either by low-pressure ammonia plasma treatment, or by attachment of 3-aminopropyltriethoxysilane (APTES) onto air plasma-treated PDMS. The resultant coatings were extensively characterized by X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), contact angle measurements, and atomic force microscopy (AFM). The results show that the impact of the plasma treatment on the physical properties on the topmost surface of the PDMS is critically important for the characteristics of the layered coatings. PMID- 19123805 TI - Effect of the supporting electrolyte anion on the thickness of PSS/PAH multilayer films and on their permeability to an electroactive probe. AB - Quartz crystal microbalance and cyclic voltammetry are used to investigate the influence of the supporting salt of polyelectrolyte solutions on the buildup and the structure of PSS/PAH polyelectrolyte multilayers (PSS: poly(4-styrene sulfonate); PAH: poly(allylamine hydrochloride)). This film constitutes a model polyelectrolyte multilayer system. The supporting electrolytes were sodium salts where the nature of the anion was changed by following the Hofmeister series from cosmotropic to chaotropic anions (F-, Cl-, NO3-, ClO4-). For all the investigated anions, the film thickness increases linearly with the number of deposition steps.Wefind that chaotropic anions lead to larger thickness increments per bilayer during the film buildup than cosmotropic ones, confirming results found on PSS/PDADMA multilayers (PDADMA:poly(diallyldimethylammonium)). Films constituted by more than nine PSS/PAH bilayers are still permeable to hexacyanoferrate(II) ions, Fe(CN)(6)4-, whatever the nature of the supporting salt anion. On the other hand, these films are impermeable to ruthenium(II) hexamine ions, Ru(NH3)(6)2+, after the third PAH layer in the presence of NaF, NaCl, or NaNO3. These results are explained by the presence of an excess of positive charges in the film, which leads to a positive Donnan potential. We find that this potential is more positive when more chaotropic anions are used during the film buildup. We also find that a film constructed in the presence of chaotropic anions swells and becomes more permeable to Fe(CN)(6)4- ions when the film is brought into contact with a solution containing more cosmotropic anions. All our experimental findings can be explained by a strong interaction between chaotropic anions with the NH3+groups of PAH that is equivalent, as far as the multilayer buildup and electrochemical response is concerned, to a deprotonation of PAH as it is observed when the film is constructed at a higher pH. We thus arrive to a coherent explanation of the effect of the nature of the anions of the supporting electrolyte on the polyelectrolyte multilayer. We also find that great care must be taken when investigating polyelectrolyte multilayer films by electrochemical probing because electrochemical reactions involving the probes can appreciably modify the multilayer structure. PMID- 19123806 TI - Toward the total synthesis of the brasilinolides: construction of a differentially protected C20-C38 segment. AB - An efficient, convergent synthesis of a differentially protected C20-C38 segment of the brasilinolides is described. Iterative 1,4-syn aldol additions and ketone reductions were employed to construct the two related stereotetrads, while a sequence of Horner-Wadsworth-Emmons (HWE) coupling, CBS reduction, and Sharpless AE installed the epoxy alcohol functionality. PMID- 19123807 TI - Assemblies of double hydrophilic block copolymers and oppositely charged dendrimers. AB - The association of poly(ethylene oxide-b-methacrylic acid) and poly(amidoamine) dendrimers was examined by dynamic light scattering and small angle neutron scattering. With increasing amounts of the G4 dendrimer as the counterion, the size of the assemblies increases until it reaches a hydrodynamic radius of about 70 nm. The structure is consistent with poly(methyl methacrylate) (PMAA) chains closely aggregating with the dendrimers at low dendrimer amounts and volume filling PMAA blocks at higher dendrimer contents. Similar behavior was observed for G4 and G2 dendrimers, while smaller G0 molecules showed an opposite dependence. The results represent an example of finite size assemblies formed by "electrostatic self-assembly" that are stable in aqueous solution and represent equilibrium structures, the structure and size of which can be tuned through the building units, loading ratio, and pH. PMID- 19123808 TI - Photochemical size reduction of CdSe and CdSe/ZnS semiconductor nanoparticles assisted by n pi* aromatic ketones. AB - A novel effect of n pi* aromatic ketones on the quantum dots optical properties is reported. By controlling experimental conditions such as time of irradiation, presence of air in the media, hydrogen donor capacity of the solvent, and irradiation wavelength, core and core-shell CdSe QDs can be resized as convenient. PMID- 19123809 TI - From monomer to bulk: appearance of the structural motif of solid iodine in small clusters. AB - Formation of iodine clusters in a solid krypton matrix was studied using resonance Raman spectroscopy with a 1 cm(-1) resolution. The clusters were produced by annealing of the solid and recognized by appearance of additional spectral transitions. Two distinct regions, red-shifted from the fundamental vibrational wavenumber of the isolated I(2) at 211 cm(-1), were observed in the signal. The intermediate region spans the range 196-208 cm(-1), and the ultimate region consists of two peaks at 181 and 190 cm(-1) nearly identical to crystalline I(2). The experimental results were compared to DFT-D level electronic structure calculations of planar (I(2))(n) clusters (n = 1-7). The dimer, trimer, and tetramer structures, where the I(2) molecule is complexed from one end, were found to exhibit vibrational shifts corresponding to the intermediate size clusters. The larger, bulklike shift appears when the iodine molecule is coordinated from two opposite directions as in the case of a pentamer and higher clusters. Starting from the pentamer, the structural motif of crystalline iodine is clearly recognized in the clusters. PMID- 19123810 TI - Protein-directed synthesis of highly fluorescent gold nanoclusters. AB - A simple, one-pot, "green" synthetic route, based on the "biomineralization" capability of a common commercially available protein, bovine serum albumin (BSA), has been developed for the preparation of highly stable Au nanocrystals (NCs) with red emission and high quantum yield. PMID- 19123811 TI - Construction of carbohydrate microarrays by using one-step, direct immobilizations of diverse unmodified glycans on solid surfaces. AB - Carbohydrate microarrays have received great attention as high-throughput analytic tools in studies of carbohydrate-mediated biological processes. Most of the methods employed to fabricate glycan microarrays rely on the immobilization of modified glycans on the properly derivatized surfaces. This immobilization strategy requires the availability of modified glycans whose syntheses in many cases are time-consuming and difficult. We have developed a simple and direct immobilization technique that involves a one-step, site-specific attachment of diverse unmodified glycans to the hydrazide-derivatized glass surface. To demonstrate the generality of this direct immobilization method, we examined its use for the construction of carbohydrate microarrays containing a variety of glycans. The results of protein and cell-binding experiments indicate that the glycan microarrays, prepared by using this methodology, are applicable to the rapid evaluation of glycan-mediated biomolecular interactions and the determination of quantitative binding affinities between carbohydrates and proteins. PMID- 19123812 TI - SAMs on gold derived from the direct adsorption of alkanethioacetates are inferior to those derived from the direct adsorption of alkanethiols. AB - Self-assembled monolayers (SAMs) on gold derived from the direct adsorption of thioacetic acid S-decyl ester (C10SAc) and thioacetic acid S-octadecyl ester (C18SAc) were compared to the corresponding SAMs derived from the analogous adsorption n-decanethiol (C10SH) and n-octadecanethiol (C18SH). All SAMs were characterized using ellipsometry, contact angle goniometry, polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS), and X-ray photoelectron spectroscopy (XPS). The comparison revealed that the SAMs generated from the thioacetates are not as densely packed and well ordered as the SAMs generated from the thiols. Furthermore, studies of the kinetics of adsorption found that the thioacetates adsorb more slowly than the corresponding thiols. PMID- 19123813 TI - Growing hardier crops for better health: Salinity tolerance and the nutritional value of broccoli. AB - To evaluate the variations in the nutritional components of a broccoli cultivar under saline stress, two different NaCl concentrations (40 and 80 mM) were assayed. Glucosinolates, phenolic compounds, and ascorbic and dehydroascorbic acids (vitamin C) were analyzed by HPLC, and mineral composition was determined by ICP spectrophotometry. Qualitative differences were observed for several bioactive compounds depending on the plant organ and the intensity of the salt stress. Glucosinolate content showed the most significant increase in the florets; phenolic compounds also increased in the florets, whereas no variation in the vitamin C content was observed as a result of the saline treatments. The mineral composition of the edible parts of the inflorescences remained within the range of the recommended values for human consumption. Overall, the nutritional quality of the edible florets of broccoli was improved under moderate saline stress. PMID- 19123814 TI - Antimicrobial activity of coffee melanoidins-a study of their metal-chelating properties. AB - Melanoidins comprise a substantial proportion of severely heat-treated foods such as baked cereals or roasted coffee and are widely consumed dietary components. The antimicrobial activity of coffee melanoidins against different pathogenic bacteria has been studied, finding that such activity is due to their metal chelating properties. Three different mechanisms have been observed: at low concentrations melanoidins exerted a bacteriostatic activity mediated by iron chelation from the culture medium; in the case of bacterial strains that are able to produce siderophores for iron acquisition, melanoidins chelate the siderophore Fe3+ complex, which could decrease the virulence of such pathogenic bacteria; and, finally, coffee melanoidins also exerted a bactericide activity at high concentrations by removing Mg2+ cations from the outer membrane, promoting the disruption of the cell membrane and allowing the release of intracellular molecules. PMID- 19123816 TI - Urea's action on hydrophobic interactions. AB - For more than a century, urea has been commonly used as an agent for denaturing proteins. However, the mechanism behind its denaturing power is still not well understood. Here we show by molecular dynamics simulations that a 7 M aqueous urea solution unfolds a chain of purely hydrophobic groups which otherwise adopts a compact structure in pure water. The unfolding process arises due to a weakening of hydrophobic interactions between the polymer groups. We also show that the attraction between two model hydrophobic plates, and graphene sheets, is reduced when urea is added to the solution. The action of urea is found to be direct, through its preferential binding to the polymer or plates. It is, therefore, acting like a surfactant capable of forming hydrogen bonds with the solvent. The preferential binding and the consequent weakened hydrophobic interactions are driven by enthalpy and are related to the difference in the strength of the attractive dispersion interactions of urea and water with the polymer chain or plate. This relation scales with square root(epsilon(b)), where epsilon(b) is the Lennard Jones (LJ) energy parameter for each group on the chain. Larger values of epsilon(b) increase the preferential binding and result in a larger decrease of the hydrophobic interactions, with a crossover at very weak dispersions. We also show that the indirect mechanism, in which urea acts as a chaotrope, is not a likely cause of urea's action as a denaturant. These findings suggest that, in denaturing proteins, urea (and perhaps other denaturants) forms stronger attractive dispersion interactions with the protein side chains and backbone than does water and, therefore, is able to dissolve the core hydrophobic region. PMID- 19123815 TI - Measurement of short-chain carbohydrates in common Australian vegetables and fruits by high-performance liquid chromatography (HPLC). AB - Fermentable oligo-, di-, and monosaccharides and polyols (FODMAPs) are short chain carbohydrates that can be poorly absorbed by the small intestine and may have a wide range of effects on gastrointestinal processes. FODMAPs include lactose, fructose in excess of glucose, fructans and fructooligosaccharides (FOS, nystose, kestose), galactooligosaccharides (GOS, raffinose, stachyose), and sugar polyols (sorbitol, mannitol). This paper describes an analytical approach based on HPLC with ELSD that quantifies the major FODMAPs in 45 vegetables and 41 fruits. Sorbitol and/or mannitol were measured in 18 vegetables (range = 0.09 2.96 g/100 g of fw), raffinose and/or stachyose in 7 vegetables (0.08-0.68 g/100 g of fw), and nystose and/or kestose in 19 vegetables (0.02-0.71 g/100 g of fw). Apple, pear, mango, clingstone peach, and watermelon all contained fructose in excess of glucose. Sorbitol was measured in 15 fruits (0.53-5.99 g/100 g of fw), mannitol was found in 2 fruits, and nystose or kestose was measured in 8 fruits. Understanding the importance of dietary FODMAPs will be greatly assisted by comprehensive food composition data. PMID- 19123817 TI - Persistence of plant DNA sequences in the blood of dairy cows fed with genetically modified (Bt176) and conventional corn silage. AB - To determine whether plant sequences, including transgenic sequences, are present in animal blood, we tested blood samples from Holstein cows fed with either Bt176 genetically modified corn or conventional corn. We used previously described sensitive real-time PCR assays targeting transgenic sequences (35S promoter and Bt176 specific junction sequence), a monocopy maize-specific sequence (ADH promoter), and two multicopy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The presence of Cry1A(b) protein in bovine blood samples was also tested using a sandwich ELISA kit. Our study shows the ability of plant nuclear and/or chloroplast DNA fragments to enter bovine blood circulation. However, maize nuclear DNA, both mono- and multicopy sequences, was less detected than chloroplast DNA, probably because the higher number of chloroplast copies and also possibly because nuclear DNA might be less protected by the nuclear membrane. Despite our data confirm the ability of small (ca.150 bp) plant DNA fragments to cross the intestinal barrier, we were unable to demonstrate clearly the presence of transgenic DNA or proteins in bovine blood. No sample tested positive with the two real-time PCR assays targeting transgenic sequences (35S promoter and Bt176 specific junction sequence). Only faint punctual positive results occurred randomly and were probably due to postsample collection or laboratory contamination or can be considered as artifact as they have never been confirmed. Our data highlight the difficulties to detect transgenic sequences in blood of dairy cows fed genetically modified corn (Bt176) silage. Those results show that in order to meet the consumers' demand of animals fed with GM products there is currently no cost-effective analytical procedure to replace documentary traceability. PMID- 19123818 TI - Immersion of lemons into imazalil mixtures heated at 50 degrees C alters the cuticle and promotes permeation of imazalil into rind wounds. AB - The influence of the immersion of lemon fruit in aqueous imazalil (IMZ) mixtures at 25 or 50 degrees C on the deposition and persistence of IMZ within the fruit rind and its effectiveness in controlling postharvest green mold, caused by Penicillium digitatum, was investigated. Concentrations of IMZ in the albedo tissue surrounding deep wounds in the rind were higher than the mean EC50 values for IMZ-resistant strains (1.0 microg *mL(-1)) after immersion of fruit into IMZ mixtures containing 25, 50, or 75 mg* L(-1) heated to 50 degrees C, but not those at 25 degrees C, where fungicide deposition was insufficient. IMZ residue, one day after treatment with 25, 50, or 75 mg* L(-1), was 3, 5, or 7 times higher after treatment at 50 degrees C compared to treatment at 25 degrees C. IMZ residues within the albedo of unwounded fruit treated with 25, 50, or 75 mg* L( 1) at 50 degrees C were 0.8, 1.3, or 2.0 microg * g(-1), respectively, while those similarly treated at 25 degrees C had negligible residues. Residues following treatment at 50 degrees C in the albedo did not decline during storage at 10 degrees C after 60 days. IMZ residues in the flavedo were higher than those in the albedo: after treatment with 25, 50, or 75 mg* L(-1) IMZ, they averaged 1.0, 1.2, or 2.7 microg * g(-1), respectively, after treatment at 25 degrees C, and 5.5, 7.9, or 16.2 microg * g(-1), respectively, after treatment at 50 degrees C. IMZ effectiveness as an eradicant to control green mold improved when it was heated. Green mold was reduced by 22 or 95% after treatment of lemons inoculated 1 h before immersion of lemons in IMZ at 50 mg* L(-1) at 25 or 50 degrees C, respectively. Green mold was reduced by 18 or 61% after treatment of lemons inoculated 1 day after immersion of lemons in IMZ at 50 mg* L(-1) at 25 or 50 degrees C, respectively. Scanning electron microscopy of the rind surface indicated that treatment at 50 degrees C for 2 min also caused ruptures in the cuticle. PMID- 19123819 TI - Thiophene dendrimers as entangled photon sensor materials. AB - The ability to do spectroscopy with a small number of entangled photons is an important development in the area of materials and sensing. This report investigates the effects of increasing thiophene dendrimer generation on the cross-section for both entangled (sigmaE) and random (deltaE) two-photon absorption cross-sections. Nonlinear optical properties of dendrimers are an interesting area of study because of potential applications in optical signal processing and remote sensing, and the use of a nonlinear optical material as a sensor for entangled photons offers great possibilities for applications in quantum lithography. Entangled two-photon absorption (ETPA) experiments and two photon excited fluorescence (TPEF) experiments vary by at least 10 orders of magnitude in the photon flux used to probe the material. ETPA cross-sections from liquid samples as well as those of thin-film samples are investigated. An increase in sigmaE and de;taR with increasing dendrimer generation is observed, suggesting that the thiophene groups within the dendrimer nonlinearly absorb in a cooperative manner, which is further evidenced in the variation of cross-section per thiophene group. The nonlinear spectroscopic features obtained by the TPEF measurements were also obtained by the ETPA experiments, despite the fact that 10 orders of magnitude fewer photons are used in the latter technique. All dendrimer generations investigated in this work are found to have great potential for applications in quantum optical devices. PMID- 19123820 TI - Repellency of an over-the-counter essential oil product in China against workers of red imported fire ants. AB - Repellency of an over-the-counter essential oil product from China, and its major components against workers of red imported fire ants, Solenopsis invicta Buren, was evaluated using an ant digging bioassay. Three concentrations (1.0, 10.0, and 100.0 mg/kg in sand) of the product were tested. At 100.0 mg/kg, the digging suppress index (DSI) was 1.0 +/- 0.00 (mean +/- SE) for all six test colonies, indicating this product produced a complete digging suppression; at 10 mg/kg, DSI was 0.22 +/- 0.089 to 0.75 +/- 0.12 and significant repellency occurred against five of six colonies; and at 1.0 mg/kg, DSI was 0.21 +/- 0.091 to 0.38 +/- 0.14 and significant repellency occurred against four of six colonies. The chemical components of this product were analyzed using gas chromatography-mass spectrometry (GC-MS). Camphor, eucalyptol, eugenol, menthol, methyl salicylate, and phenylethanol were identified. A digging bioassay was also conducted on each of those identified compounds at concentrations of 1.0, 10.0, and 100.0 mg/kg. Based on pooled data from three colonies, each component significantly suppressed the digging behavior at 100 mg/kg. Eugenol, menthol, and methyl salicylate significantly suppressed the digging at 10 mg/kg. At 1.0 and 10.0 mg/kg, DSI for eucalyptol was -0.039 +/- 0.032 and -0.050 +/- 0.021, respectively. The negative DSI indicated a digging facilitation. However, only at 10.0 mg/kg, was such facilitation statistically significant. PMID- 19123821 TI - Fatty acid and stable carbon isotope characterization of Camelina sativa oil: implications for authentication. AB - The importance of authenticity characterization is an increasing and pressing requirement for all foods. Vegetable oil is one of the most studied foods because of its nutritional and medicinal properties in a correct diet. In this study, a total of 53 Camelina sativa samples, from all known growing areas, were chemically and isotopically characterized. The fatty acid content of camelina oil was determined by gas chromatography (GC), and the ratios of stable carbon isotopes ((13)C/(12)C) of individual fatty acids and seed/bulk oil were determined by gas chromatography-combustion-stable isotope ratio mass spectrometry (GC/C/IRMS) and elemental analysis-stable isotope ratio mass spectrometry (IRMS). A total of 17 different fatty acids were detected by GC, with omega3 R-linolenic acid (C(18:3n3)) being the most abundant (29.7-40.0 wt %). Oleic acid (C(18:1n9)), linoleic acid (C(18:2n6)) and eicosenoic acid (C(20:1n9)) all belong to the second group of major fatty acids. The stable carbon isotopic values (delta(13)C) fell into a range typical for C(3) plants. The use of delta(13)C(18:2n6) vs delta(13)C(18:3n3) correlation could show cases where impurity or adulteration is suspected, whereas principal component analysis clearly separates oil samples from different continents. Preliminary results on the camelina oil authentication procedure provide a basis for the investigation of geographical origin and the further distinction between camelina and camelina refined or other, less expensive oils. PMID- 19123823 TI - Solvent dependence of conformational distribution, molecular geometry, and electronic structure in adenosine. AB - Solvation dynamics of adenosine in water and chloroform solvents under ambient conditions has been investigated using both force-field molecular dynamics (MD) and first-principles Car-Parrinello molecular dynamics (CPMD) calculations. First, the solvent dependence of the equilibria between anti-syn forms, C((3')) endo-C((2'))-endo conformations, and carbinol group rotamers has been discussed from MD calculations. We find that in both the solvents the adenosine molecule can remain either in anti or syn conformations. But, the anti-syn interconversion occurs relatively faster in water solvent than in chloroform solvent. Because of the relatively larger time scale for the interconversion, anti and syn conformational states of adenosine are studied separately in water and chloroform solvents using CPMD calculations. The dipole moments calculated from CPMD and MD calculations for adenosine in water are significantly larger than in chloroform solvent. On the basis of the CPMD calculations, the syn form of adenosine in water has a larger dipole moment than the anti form. Moreover, the molecular geometry of anti and syn forms of adenosine in these two solvents is reported. We report a remarkable solvent effect on the geometry of the anti form of the adenosine, which is attributed to differences in the intermolecular and intramolecular hydrogen-bonding stabilization. We also report the solvent effect on the frontier Kohn-Sham orbitals and energy gaps for anti-syn conformational states. Finally, we report the solvation shell structure of adenosine in both the solvents, and we find that the solvent-solute interaction is site-specific in the case of water while in chloroform solvent the interaction is globular isotropic in nature. PMID- 19123824 TI - Molecular simulation of ion transport in silica nanopores. AB - Ion distribution and transport of KCl aqueous solutions at the junction of hydrophobic and hydrophilic regions inside silica nanopores were studied by using two kinds of molecular simulation: grand canonical Monte Carlo (GCMC) simulations and nonequilibrium molecular dynamics (NEMD) simulations. The nanopores were 2 nm diameter silica pores in which surface functional groups, -SiOH, had been modified by hydrophobic surface functional groups, -SiCH(3), within three different lengths along the pore direction (z-direction), L(z0) = 0, 2, and 4 nm. If L(z0) is long enough, water could not enter the hydrophobic region, but for all L(z0) studied here, water entered the hydrophobic region. When an external electric field was applied along the z-direction, ions could not pass through the hydrophobic region when the external electric field was less than a threshold level, E(0), whereas the ionic current increased relatively linearly with increasing electric field strength above E(0). In 2 nm diameter fluidic pores, the electrical potential barrier appeared at the junction between the hydrophilic and hydrophobic regions due to the difference in dipole moment of the surface functional groups, although the overall surface charge of the pore was neutral. This junction formed an electrical potential threshold, and the ionic current could be modulated by adjusting the external electric field. PMID- 19123825 TI - Stability and rupture of archaebacterial cell membrane: a model study. AB - It is known that the thermoacidophilic archaebacterium Sulfolobus acidocaldarius can grow in hot springs at 65-80 degrees C and live in acidic environments (pH 2 3); however, the origin of its unusual thermal stability remains unclear. In this work, using a vesicle as a model, we study the thermal stability and rupture of archaebacterial cell membrane. We perform a simulation investigation of the structure-property relationship of monolayer membrane formed by bolaform lipids and compare it with that of bilayer membrane formed by monopolar lipids. The origin of the unusually thermal stability of archaebacterial cell and the mechanism for its rupture are presented in molecular details. PMID- 19123826 TI - Femtosecond solvation dynamics in a micron-sized aggregate of an ionic liquid and P123 triblock copolymer. AB - Dynamic light scattering studies indicate that addition of a room temperature ionic liquid (RTIL, [pmim][Br]), to a triblock copolymer (P123) micelle leads to the formation of giant P123-RTIL clusters of size (diameter) 40 nm in 0.9 M and 3500 nm (3.5 microm) in 3 M RTIL. They are much larger than a P123 micelle ( approximately 18 nm) or [pmim][Br] (1.3 nm). Dynamics in different regions of the P123-RTIL aggregate is probed by variation of the excitation wavelength (lambda(ex)) using femtosecond up-conversion. For lambda(ex) = 375 nm, the nonpolar core of the P123-RTIL aggregate is preferentially excited while lambda(ex) = 435 nm selects the polar corona region. Solvation dynamics and anisotropy decay of coumarin 480 (C480) in a P123-RTIL giant aggregate are markedly different from those in either P123 micelle or those in an aqueous solution of the RTIL. For lambda(ex) = 405 nm in 5 wt % P123 and 0.9 M RTIL average rotational time, ( = 1350 ps) of C480 is approximately 7 times longer than that (200 ps) in an aqueous solution of the RTIL in the absence of P123 and is shorter than that (3000 ps) in a P123 micelle. In 0.9 M RTIL and 5 wt % P123, solvation dynamics in the corona region (lambda(ex) = 435 nm, = 75 ps) is approximately 25 times faster than that at the core region (at lambda(ex) = 375 nm, = 1900 ps). The solvation dynamics in the core of the P123-RTIL aggregate is faster than that in P123 micelle (3550 ps in the core) and is much slower than that (130 ps) in an aqueous solution containing 0.9 M RTIL. In the 3.5 microm sized aggregate (3 M RTIL and P123), the solvation dynamics in the core ( = 500 ps) is approximately 4 times faster than that in 0.9 M RTIL. PMID- 19123827 TI - Effect of surfactant mixing on partitioning of model hydrophobic drug, naproxen, between aqueous and micellar phases. AB - Mixed surfactants may improve the performance of surfactant-enhanced solubilization of drugs and thus can serve as the tool for increased bioavalaibility, controlled drug release, and targeted delivery. Solubilization of Naproxen by micellar solutions at 25 degrees C using single and mixed surfactant systems was measured and compared. Solubilization capacity determined with spectrophotometry and tensiometry has been quantified in terms of molar solubilization ratio, micelle-water partition coefficient, and locus of solubilization. Cationic surfactants exhibited higher solubilization capacity than nonionics and anionics, the efficiency increasing with chain length. Mixing effect of surfactants on mixed micelle formation and solubilization efficiency has been discussed in light of regular solution approximation (RSA). Equimolar cationic-nonionic surfactant combinations showed better solubilization capacity than pure cationics or nonionics, which, in general, increased with increase in hydrophobic chain length. Equimolar cationic-nonionic-nonionic ternary surfactant systems exhibited intermediate solubilization efficiency between their single and binary counterparts. Use of RSA has been extended, with fair success, to predict the partition coefficient of ternary surfactant systems using data from binary mixtures. The theoretical micelle-water partition coefficients calculated from the geometric mean equation compared well with experimental values. Locus of solubilization of NAP in different micellar solutions was probed by UV-visible spectroscopy. PMID- 19123828 TI - Molecular dynamics simulations of the chromophore binding site of Deinococcus radiodurans bacteriophytochrome using new force field parameters for the phytochromobilin chromophore. AB - The conformational flexibility of the tetrapyrrolic phytochromobilin (PPhiB) chromophore of the bacteriophytochrome Deinococcus radiodurans (DrCBD) in the Pr state has been investigated by molecular dynamics simulations. Because these simulations require accurate force field parameters for the prosthetic group, in the present work we developed new empirical force field parameters for the PPhiB molecule that are compatible with the CHARMM22 force field for proteins. For this reason, the new force field parameters for the nonbonded (partial atomic charges) and bonded (bonds, angles, dihedrals, improper) energy terms were derived by reproducing ab initio target data following the methodology used in the development of the CHARMM22 force field. This new set of parameters was employed to analyze structural and dynamical features of PPhiB inside DrCBD. The 45 ns all atom molecular dynamics (MD) simulation reveals the existence of two stable conformational states of the chromophore characterized by distinct torsional angles around the C-C bond at the methine bridge connecting rings A and B of the tetrapyrrole. This result supports experimental observations derived from NMR and resonance Raman spectroscopy. Furthermore, statistical analysis of H-bonding events allowed us to identify (a) important H-bonds between the propionic side chains of the chromophore and the apoprotein which may be relevant for the signal transduction step during the photoinduced cycle and (b) a network of eight water molecules which remain in the vicinity of the chromophore during the entire 45 ns production run. PMID- 19123829 TI - SO(2) saturation of the room temperature ionic liquid [C(2)mim][NTf(2)] much reduces the activation energy for diffusion. AB - The physical effect of high concentrations of reversibly dissolved SO(2) on [C(2)mim][NTf(2)] was examined using cyclic voltammetry, chronoamperometry, and ESR spectroscopy. Cyclic voltammetry of the oxidation of solutions of ferrocene, N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), and chloride in the room temperature ionic liquid (RTIL) 1-ethyl-3-methylimidazolium bis(trifluoromethanesufonyl)imide ([C(2)mim][NTf(2)]) reveals an increase in limiting current of each species corresponding to the addition of increasing concentrations of sulfur dioxide. Quantitative chronoamperometry reveals an increase in each species' diffusion coefficient with SO(2) concentration. When chronoamperometric data were obtained for ferrocene in [C(2)mim][NTf(2)] at a range of temperatures, the translational diffusion activation energy (29.0 +/- 0.5 kJ mol(- 1)) was found to be in good agreement with previous studies. Adding SO(2) results in apparent near-activationless translational diffusion. A significant decrease in the activation energy of rotational diffusion with the SO(2) saturation of a 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) solution in [C(2)mim][NTf(2)] (29.9 +/- 2.0 to 7.7 +/- 5.3 kJ mol(- 1)) was observed using electron spin resonance (ESR) spectroscopy. The reversible physical absorption of SO(2) by [C(2)mim][NTf(2)] should have no adverse effect on the ability of that ionic liquid to be employed as a solvent in an electrochemical gas sensor, and it is possible that the SO(2)-mediated reduction of RTIL viscosity could have intrinsic utility. PMID- 19123830 TI - Adsorption isotherms of water on mica: redistribution and film growth. AB - Adsorption isotherms of water on muscovite mica are obtained using grand canonical Monte Carlo simulations over a wide range of relative vapor pressures, p/p(0) at 298 K. Three distinct stages are observed in the adsorption isotherm. A sharp rise in the water coverage occurs for 0 < p/p(0) < 0.1. This is followed by a relatively slow increase in the coverage for 0.1 < or = p/p(0) < or = 0.7. Above p/p(0) = 0.7, a second increase in the coverage occurs due to the adsorption of water with bulklike features. The derived film thickness and isotherm shape for the simple point charge (SPC) water model is in excellent agreement with recent experiments of Balmer et al. [ Langmuir 2008 , 24 , 1566 ]. A novel observation is the significant redistribution of water between adsorbed layers as the water film develops. This redistribution is most pronounced for 0.1 < or = p/p(0) < or = 0.7, where water is depleted from the inner layers and film growth is initiated on the outer layer. During this stage, potassium hydration is found to play a dominant role in the rearrangement of water near the mica surface. The analysis of structural features reveals a strongly bound first layer of water molecules occupying the ditrigonal cavities between the potassium ions. In-plane structure of oxygen in the second layer, which forms part of the first hydration shell of potassium, reveals a liquidlike structure with the oxygen oxygen pair correlation function displaying features similar to bulk water. Isosteric heats of adsorption were found to be in good agreement with the differential microcalorimetric data of Rakhmatkariev ( Clays Clay Miner. 2006 , 54 , 402 ), over the entire range of pressures investigated. Both SPC and extended simple point charge (SPC/E) water models were found to yield qualitatively similar adsorption and structural characteristics, with the SPC/E model predicting lower coverages than the SPC model for p/p(0) > 0.7. PMID- 19123831 TI - Gas-phase reactions of sulfur hexafluoride with transition metal and main group atomic cations: room-temperature kinetics and periodicities in reactivity. AB - Gas-phase reactions of SF(6) were investigated with 46 different atomic metal and main group cations at room temperature using an Inductively-Coupled Plasma/Selected-Ion Flow Tube (ICP/SIFT) tandem mass spectrometer. The atomic ions were produced at about 5500 K in the ICP source and allowed to decay radiatively and to thermalize by collisions with argon and helium atoms prior to reaction downstream in a flow tube in helium buffer gas at 0.35 +/- 0.01 Torr and 295 +/- 2 K. Rate coefficients and product distributions were measured for the reactions of fourth-row atomic ions from K(+) to Se(+), of fifth-row atomic ions from Rb(+) to Te(+) (excluding Tc(+)), and of sixth-row atomic ions from Cs(+) to Bi(+). The early transition metal ions react with SF(6) very efficiently (k/k(c) = 0.56-0.96) to produce MF(m)(+) (m = 1-4) and SF(n)(+) (n = 1-4) ions, whereas the late transition metal ions react much less efficiently (k/k(c) < 0.2) to form M(+)(SF(6)) adduct ions. Reactions of SF(6) with Ca(+), Sr(+), Ba(+), Ge(+), and As(+) proceed efficiently (k/k(c) = 0.35-0.85) through various channels, while all other main group metal ions are inert toward sulfur hexafluoride. Primary and secondary adduct formation was observed to exhibit equilibrium kinetics, and the standard free energy change for SF(6) addition is found to correlate with the efficiency of addition according to log[k/k(c)] = -8.7 + 7.8 log[-DeltaG(o)/(kcal mol(-1))]. Several MF(m)(+) ions were observed to react further with SF(6) to produce MF(m+k)(+), SF(n)(+), and MF(m)(+)(SF(6)) as secondary products. PMID- 19123832 TI - On the mechanism of iron(III)-dependent oxidative dehydrogenation of amines. AB - Kinetic and structural data are presented for the iron-promoted dehydrogenation of the amine, [Fe(III)L3]3+ (1), L3 = 1,9-bis(2'-pyridyl)-5-[(ethoxy-2'' pyridyl)methyl]-2,5,8-triazanonane. Spectroscopic and electrochemical experiments under the exclusion of dioxygen helped to identify reaction intermediates and the final product, the Fe(II)-monoimine complex [Fe(II)L4]2+ (2), L4 = 1,9-bis(2' pyridyl)-5-[(ethoxy-2''-pyridyl)methyl]-2,5,8-triazanon-1-ene. 2 is formed by disproportionation of the starting complex 1 by a three-step reaction mechanism, most likely via ligand-centered radical intermediates. The rate law can be described by the second-order rate equation, -d[(Fe(III)L3)3+]/dt = k(EtO)- [(Fe(III)L3)3+][EtO-], with k(EtO)- = 4.92 +/- 0.01 x 104 M(-1) s(-1) (60 degrees C, mu = 0.01 M). The detection of general base catalysis and a primary kinetic isotope effect (k(EtO)-(H)/k(EtO)-(D) = 1.73) represents the first kinetic demonstration that the deprotonation becomes rate determining followed by electron transfer in the oxidative dehydrogenation mechanism. We also isolated the Fe(II)-monoimine complex 2 and determined its structure in solution (NMR) and in the solid state (X-ray). PMID- 19123833 TI - An efficient synthesis of terminal allenes from terminal 1-alkynes. AB - We have developed a modified method for the synthesis of terminal allenes from terminal 1-alkyne: The reaction of 1-alkynes with 1.8 equiv of Cy(2)NH and 2.5 equiv of paraformaldehyde mediated by CuI (0.5 equiv) in refluxing dioxane may produce terminal allenes in much higher yields than the previously reported protocol and many functional groups such as mesylate, hydroxyl group, ether, amide, etc. may be tolerated. PMID- 19123834 TI - Kinetics and mechanism of the sphere-to-rod transition of triblock copolymer micelles in aqueous solutions. AB - The kinetics of the sphere-to-rod transition of micelles composed of triblock copolymers of ethylene oxide and propylene oxide (EO(20)PO(70)EO(20)) have been investigated using dynamic light scattering (DLS) and cryogenic electron transmission microscopy (Cryo-EM). Sphere-to-rod transition is induced by a solvent jump, initiated by adding KCl and ethanol to an aqueous micellar solution. The growth process of the wormlike micelles depends on the experimental conditions and has two distinct regions that can be described as initiation period and actual growth to equilibrium. All growth curves exhibit a single relaxation time that represents the lifetime of the micelles. The growth curves collapse into a master curve, when shifted by the relaxation time, indicating that the actual growth process of the micelles in all samples occurs through the same mechanism. The relaxation time decreases with increasing surfactant concentration. Additionally, some of the formed micelles exhibit a caterpillarlike shape in which some of the original spherical species can still be detected. These facts suggest that the micelles grow longer predominantly by random coagulation/fragmentation reactions involving micellar species of different sizes. However, the appearance of a unimer peak is detected with DLS during the growth stage. This implies that unimer exchange may also contribute to the elongation of the micelles. PMID- 19123835 TI - Mechanism of hydrogen peroxide production by copper-bound amyloid beta peptide: a theoretical study. AB - The amyloid beta peptide (Abeta) of Alzheimer's disease evolves hydrogen peroxide in vitro in the presence of Cu(II), external reducing agents, and molecular oxygen, without producing detectable amounts of the one-electron reduced intermediate, superoxide, O(2)(-*). The mechanism of this process was examined by ab initio computational chemistry techniques in systems that model the binding of Cu(II) to the His13His14 fragment of Abeta. The catalytic cycle begins with the reduction of the most stable Cu(II) complex to the most stable Cu(I) complex. This Cu(I) complex forms a Cu(II)-like adduct with (3)O(2) that cannot dissociate in water to yield O(2)(-*). However, it can be reduced by proton-coupled electron transfer to an adduct between HOO(-) and the Cu(II)-like complex, which in turn can be protonated. The protonated complex decomposes to yield H(2)O(2) by an associative-dissociative mechanism, thus completing the cycle. PMID- 19123836 TI - Brevisin: an aberrant polycyclic ether structure from the dinoflagellate Karenia brevis and its implications for polyether assembly. AB - Brevisin is an unprecedented polycyclic ether isolated from the dinoflagellate Karenia brevis, an organism well-known to produce complex polycyclic ethers. The structure of brevisin was determined by detailed analyses of MS and 2D NMR spectra and is remarkable in that it consists of two separate fused polyether ring assemblies linked by a methylene group. One of the polycyclic moieties contains a conjugated aldehyde side chain similar to that recently observed in other K. brevis metabolites, though the "interrupted" polyether structure of brevisin is novel and provides further insight into the biogenesis of such fused ring polyether systems. On the basis of the unusual structure of brevisin, principles underlying the initiation of polyether assemblies are proposed. Brevisin was found to inhibit the binding of [(3)H]-PbTx-3 to its binding site on the voltage-sensitive sodium channels in rat brain synaptosomes. PMID- 19123837 TI - Effect of additives on chemoselectivity and diastereoselectivity in the catalytic epoxidation of chiral allylic alcohols with hydrogen peroxide and binuclear manganese complexes. AB - The catalytic oxidations of chiral allylic alcohols 2 by manganese complexes of the cyclic triamine 1,4,7-trimethyl-1,4,7-triazacyclononane (tmtacn) 1 and hydrogen peroxide as oxygen donor in the presence of co-catalyst are investigated to understand the factors that affect the catalyst selectivity. Chemoselectivity and diastereoselectivity of catalyst 1 are significantly affected by the structure of the allylic alcohol and the nature and amount of co-catalyst. More pronounced is the influence of the amount of added molar equivalents of H(2)O(2) (20-110 mol % with respect to the substrate). Our present results reflect the complex redox chemistry of the Mn catalyst 1/H(2)O(2)/co-catalyst system in the early phase of the alkene oxidation. PMID- 19123838 TI - Proton-transfer reactions of nitroalkanes: the role of aci-nitro species. AB - Proton-transfer reactions of two systems, ionization of a series of small carbon acids in water (the Pearson system) and reactions of substituted phenylnitromethanes, were examined in detail computationally. Comparison of experimental reactivity and pK(a) with calculated relative activation barrier and reaction energy for the Pearson system suggested that the origin of the well-know nitroalkane anomaly does not reside in the reactivity but in the equilibrium. For the reactions of substituted phenylnitromethanes, proton transfers among three species, PhCH(2)NO(2), PhCHNO(2)(-), and PhCH=NO(2)H, were examined, and the role of the aci-nitro species (PhCH=NO(2)H) was evaluated on the basis of its stability and reactivity. Protonation of PhCHNO(2)(-) by H(2)O was suggested to occur kinetically on the oxygen site, but due to its instability PhCH=NO(2)H does not contribute to the overall reaction energetics. The protonation of PhCHNO(2)( ) under acidic conditions occurs on the oxygen site to give PhCH=NO(2)H both kinetically and thermodynamically. The aci-nitro species thus formed appears to give PhCH(2)NO(2) via intramolecular H(2)O-mediated proton transfer, but a possibility of the route through PhCHNO(2)(-)-C-protonation would not be fully eliminated. PMID- 19123840 TI - Instantaneous deprotection of tosylamides and esters with SmI(2)/amine/water. AB - SmI(2)/amine/water mediates instantaneous cleavage of tosyl amides and tosyl esters. Highly hindered, sensitive and functionalized substrates were successfully deprotected in near quantitative yield. PMID- 19123841 TI - Order-disorder phase transformation of triacylglycerols: effect of the structure of the aliphatic chains. AB - Plant oils have been used as environmentally benign lubricants since they present high viscosity index and flash points and low evaporation loss. Triacylglycerols (TAG) are the major components of naturally occurring oils and fats and are able to produce high strength lubricant films. One of the main concerns that hinders the usage of triacylglycerols as lubricants, however, is the thermal stability of these molecules. In this paper, we report on the effect of chain structure on density, viscosity, and thermal stability of triacylglycerols using molecular dynamics simulations. The selected triacylglycerols are trilauroylglycerol (LLL TAG), tristearoylglycerol (SSS-TAG), trans-trioleoylglycerol (trans-OOO-TAG), and trans-trilinolenoylglycerol (trans-LeLeLe-TAG). The first two TAGs are saturated molecules with a different number of carbons in the chain, and the second two TAGs are monounsaturated and polyunsaturated molecules, respectively. The computed results demonstrate that the length of the aliphatic chain influences the physical properties of triacylglycerols. TAGs with short chain (LLL-TAG) show higher density than TAGs with longer chains. Viscosity is determined by the degree of recoil of the aliphatic chains and by the number and location of unsaturated bonds. Thermal stability, as represented by the ability of triacylglycerols to stay in a disordered phase during the cooling process, is related to the order-disorder phase transition temperature. Since the phase transition temperature can be correlated to the thermal stability during the cooling process, LeLeLe-TAG shows the highest thermal stability among the systems considered. These results can aid in the design of molecules with specific lubrication properties. PMID- 19123842 TI - Total synthesis of (-)-platensimycin, a novel antibacterial agent. AB - An enantioselective synthesis of platensimycin, a novel antibiotic natural product that inhibits bacterial beta-ketoacyl-(acyl-carrier-protein) synthase (FabF), is described. Our synthetic strategy for the construction of the oxatetracyclic core involved an intramolecular Diels-Alder reaction. Our preliminary studies provided a complex tetracyclic product by first undergoing an interesting 1,5-hydride shift followed by a Diels-Alder reaction. Further optimization of the diene's electronic properties, by incorporation of a methoxy group, led to the oxatetracyclic core of platensimycin. The three required chiral centers, including two all-carbon quaternary chiral centers, were built in the intramolecular Diels-Alder step. The synthesis utilized natural (+)-carvone as the key chiral starting material, which determined the stereochemistry of the final product. The synthesis also featured an efficient Petasis olefination, a hydroboration sequence, a Gais's asymmetric Horner-Wadsworth-Emmons reaction, and a mercury salt catalyzed enol ether isomerization. PMID- 19123843 TI - Synthesis of mannose and galactose oligonucleotide conjugates by bi-click chemistry. AB - Glyco oligonucleotide conjugates, each exhibiting two mannose and two galactose residues, were efficiently synthesized by two successive 1,3-dipolar cycloadditions (click chemistry). Two phosphoramidite derivatives were used: one bearing a bromoalkyl group as a precursor to azide functionalization and another bearing a propargyl group. After a first cycloaddition with a mannosyl-azide derivative, the bromine atoms were substituted with NaN(3) and a second click reaction was performed with a 1'-O-propargyl galactose, affording the heteroglyco oligonucleotide conjugate. PMID- 19123844 TI - Flexibility of the saturated five-membered ring in 2,5-pyrrolidinedione (succinimide): electron diffraction and quantum-chemical studies with use of vibrational spectroscopy data. AB - The flexibility of succinimide molecule has been studied for the first time by quantum-chemical (at the MP2 level with up to the 6-311G(3df,2p) basis sets) and gas-phase electron diffraction (GED) methods using vibrational spectroscopy data from literature. The analysis of vibrational spectra, performed for the molecular model of C(2v) symmetry (predicted by high-level ab initio calculations) using the scaling procedure, has shown that the two out-of-plane ring motions, that is, ring-bending and ring-twisting, are practically pure modes of different symmetry types and can be considered separately. The one-dimensional potential curves for the ring-bending and ring-twisting vibrations calculated at the MP2(full)/6 311G(3df,2p) level could be approximated by harmonic and anharmonic functions, respectively. The diverged energy levels for the ring-twisting vibration and the constant transition frequencies for the bending motion, obtained by the solution of the direct one-dimensional problem for the nonrigid model, demonstrate this statement. In the GED analysis, the succinimide molecule with a large-amplitude ring-twisting motion was described by a dynamic model with the distribution of pseudoconformers according to the calculated potential function taking into account structural relaxation effects from the MP2(full)/6-311G(3df,2p) calculations. This model greatly improved the fit of the GED intensities (R factor decreased from 4.6% for static model to 2.8%). The equilibrium molecular parameters r(e) determined in the dynamic approximation are very close to the corresponding values from the ab initio calculations. At the same time, the parameters of the -CH(2)-CH(2)- fragment involved in the ring-twisting motion deviate considerably from those obtained for the static model (C-C bond length by 0.027 A, =C-C-H, C-C-H, and H-C-C-N angles by up to 7 degrees). The flexibility influence on the C-C bond length is several times larger than the calculated vibrational correction (r(e) - r(a)) as well as the experimental uncertainty. PMID- 19123845 TI - In situ IR spectroscopy and ab initio calculations to study polymer swelling by supercritical CO(2). AB - The CO(2) sorption and polymer swelling of hydroxytelechelic polybutadiene (HTPB) and poly(ethylene glycol) (PEG) have been investigated as a function of temperature and CO(2) pressure by combining in situ near-infrared spectroscopy with molecular modeling. The results reported here for the PEG-CO(2) system are in a very good agreement with literature data hence validating our experimental procedure. It has been found that CO(2) sorption and swelling effect is more important for PEG than for HTPB. For both polymers, an increase of temperature leads to a strong decrease of both the CO(2) sorption and swelling. In order to identify at a molecular level the nature and strength of intermolecular interaction occurring between CO(2) and the polymers, ab initio calculations have been performed on model structures, representative of the main functional group of the polymer, and their complex with CO(2). Trans-3-hexene (3-Hex), propyl methyl ether (PME) and methoxytrimethylsilane (MTMS) have been selected to mimic the functional groups of HTPB, PEG and polydimethyl siloxane (PDMS), respectively. The last system has been chosen since previous works on the swelling of PDMS by high pressure CO(2) have revealed the high ability of CO(2) to swell both uncrosslinked and crosslinked PDMS. The calculated stabilization energies of the MTMS-CO(2), PME-CO(2), and 3-Hex-CO(2) dimers indicate that CO(2) interacts specifically with the three moieties through a Lewis acid-Lewis base type of interaction with the energies displaying the following order: E(MTMS CO(2)) = -3.59 > E(PME-CO(2)) = -3.43 > E(3-Hex-CO(2)) = -2.5 kcal/mol. Since the solubility of CO(2) in the corresponding homopolymers follows the same order, it is evidenced that the stronger the interaction between CO(2) and the polymer, the higher the CO(2) sorption. Therefore, even if one cannot exclude the influence of free volume and chain flexibility of the polymer, it appears that the solubility of CO(2) in the polymer is predominantly governed by the interaction between CO(2) and the polymer. Although the same trend is observed for the swelling of the polymer as a function of the CO(2) pressure, we have found that for a given value of CO(2) sorption, the swelling of the polymer depends on its nature, meaning that the swelling is not only governed by the CO(2)-polymer interaction but also by other intrinsic properties of the polymer. PMID- 19123846 TI - Probing the lactose.GM3 carbohydrate-carbohydrate interaction with glycodendrimers. AB - Multivalent glycoconjugates were prepared using generation-4 PAMAM dendrimers, and their interaction with Langmuir monolayers containing GM3 was investigated. Excessive carbohydrate valency adversely affects the carbohydrate-carbohydrate interaction. The GM3 monolayer selectively interacts with lactose-functionalized dendrimers in the presence of calcium ions. PMID- 19123847 TI - Photoarylation/alkylation of bromonaphthols. AB - The photochemistry of 6-bromo-2-naphthols has been studied in acetonitrile, aqueous acetonitrile, and isopropyl alcohol in the absence and in the presence of triethylamine by product distribution analysis, laser flash photolysis (LFP), fluorescence, phosphorescence, electrochemical measurements, and DFT calculations. Hydrobromic acid loss in the presence of Et(3)N occurs from the triplet state of 6-bromo-2-naphthol, generating an electrophilic carbene intermediate, which has been successfully trapped by oxygen, allyltrimethylsilane, 2,3-dimethylbut-2-ene, pyrrole, acrylonitrile, 1,4 dimethoxybenzene, and also pyridine. The generation and the reactivity of a triplet carbene intermediate has been supported by LFP, with the detection of 2,6 naphthoquinone-O-oxide (530 < lambda < 650 nm) in the presence of O(2). The electrophilic diradical character of the carbene has been supported by DFT calculations, using the B3LYP, PBE0, and MPWB1K functionals, with the 6-31+G(d,p) basis set and PCM solvation model. PMID- 19123848 TI - CuX2-mediated halolactonization reaction of monoesters of 1,2-allenyl phosphonic acids and their Suzuki cross-coupling reaction. AB - CuX(2)-mediated (X = Cl, Br) halolactonization of monoesters of 1,2-allenyl phosphonic acids is presented. The reaction proceeded smoothly under the mild condition for differently substituted allenic substrates giving the 4-halo-2,5 dihydro[1,2]oxaphosphole 2-oxides in good yields. The Suzuki cross-coupling reaction of these bromides and even chlorides with organic boronic acids under the catalysis of PdCl(2)(Sphos)(2) afforded 4-substituted-2,5 dihydro[1,2]oxaphosphole 2-oxides in moderate to good yields. PMID- 19123849 TI - Reductive defluorination of aqueous perfluorinated alkyl surfactants: effects of ionic headgroup and chain length. AB - Perfluorinated chemicals (PFCs) are distributed throughout the environment. In the case of perfluorinated alkyl carboxylates and sulfonates, they can be classified as persistent organic pollutants since they are resistant to environmentally relevant reduction, oxidation, and hydrolytic processes. With this in mind, we report on the reductive defluorination of perfluorobutanoate, PFBA (C(3)F(7)CO(2)(-)), perfluorohexanoate, PFHA (C(5)F(11)CO(2)(-)), perfluorooctanoate, PFOA (C(7)F(15)CO(2)(-)), perfluorobutane sulfonate, PFBS (C(4)F(9)SO(3)(-)), perfluorohexane sulfonate, PFHS (C(6)F(13)SO(3)(-)), and perfluorooctane sulfonate, PFOS (C(8)F(17)SO(3)(-)) by aquated electrons, e(aq)( ), that are generated from the UV photolysis (lambda = 254 nm) of iodide. The ionic headgroup (-SO(3)(-) vs -CO(2)(-)) has a significant effect on the reduction kinetics and extent of defluorination (F index = -[F( )](produced)/[PFC](degraded)). Perfluoroalkylsulfonate reduction kinetics and the F index increase linearly with increasing chain length. In contrast, perfluoroalkylcarboxylate chain length appears to have a negligible effect on the observed kinetics and the F index. H/F ratios in the gaseous fluoro-organic products are consistent with measured F indexes. Incomplete defluorination of the gaseous products suggests a reductive cleavage of the ionic headgroup occurs before complete defluorination. Detailed mechanisms involving initiation by aquated electrons are proposed. PMID- 19123851 TI - Diammoniosilane: computational prediction of the thermodynamic properties of a potential chemical hydrogen storage system. AB - Atomization energies at 0 K and heats of formation at 0 and 298 K are predicted for diammoniosilane, H(4)Si(NH(3))(2), and its dehydrogenated derivates at the CCSD(T) and G3(MP2) levels. To achieve near chemical accuracy (+/-1 kcal/mol), three corrections were added to the complete basis set binding energies based on frozen core coupled cluster theory energies: a correction for core-valence effects, a correction for scalar relativistic effects, and a correction for first order atomic spin-orbit effects. Vibrational zero-point energies were computed at the CCSD(T) or MP2 levels. The edge inversion barrier of silane is predicted to be 88.9 kcal/mol at 298 K at the CCSD(T) level and a substantial amount, -63.6 kcal/mol, is recovered upon complexation with 2 NH(3) molecules, so that the diammoniosilane complex is only 25.6 kcal/mol at 298 K above the separated reactants SiH(4) + 2NH(3). The complex is a metastable species characterized by all real frequencies at the MP2/aV(T+d)Z level. We predict the heat of reaction for the sequential dehydrogenation of diammoniosilane to yield H(3)Si(NH(2))(NH(3)) and H(2)Si(NH(2))(2) to be exothermic by 33.6 and 12.2 kcal/mol at 298 K at the CCSD(T) level, respectively. The cumulative dehydrogenation reaction yielding two molecules of hydrogen at 298 K is -45.8 kcal/mol at the CCSD(T) level. The sequential release of H(2) from H(2)Si(NH(2))(2) consequently yielding HN=SiH(NH(2)) and HN=Si=NH are predicted to be largely endothermic reactions at 45.3 and 55.7 kcal/mol at the CCSD(T) level at 298 K. If the endothermic reaction for the third step and the exothermic reactions for the release of the first two H(2) were coupled effectively, loss of three H(2) molecules from H(4)Si(NH(3))(2) would be almost thermoneutral at 0 K. PMID- 19123850 TI - A coupled polarization-matrix inversion and iteration approach for accelerating the dipole convergence in a polarizable potential function. AB - A coupled polarization-matrix inversion and iteration (CPII) method is described to achieve and accelerate the convergence of induced dipoles for condensed phase systems employing polarizable intermolecular potential functions (PIPF). The present PIPF is based on the Thole interaction dipole model in which all atomic pair interactions are considered, including those that are directly bonded covalently. Although induced dipoles can be obtained both by inverting a 3N x 3N polarization-matrix where N is the number of polarizable sites, or by a direct iterative approach, the latter approach is more efficient computationally for large systems in molecular dynamics simulations. It was found that induced dipole moments failed to converge in the direct iterative approach if 1-2, 1-3, and 1-4 intramolecular interactions are included in the Thole model. However, it is necessary to include all intramolecular interactions in the Thole model to yield the correct molecular anisotropic polarizability tensor. To solve this numerical stability problem, we reformulated the Thole interaction dipole model in terms of molecular block matrices, which naturally leads to a coupled, preconditioning algorithm that involves a polarization-matrix inversion term to account for intramolecular interactions, and an iterative procedure to incorporate the mutual polarization effects between different molecules. The CPII method is illustrated by applying to cubic boxes of water and NMA molecules as well as an alanine pentapeptide configuration, and it was shown that the CPII method can achieve convergence for the dipole induction polarization rapidly in all cases, whereas the direct iterative approach failed to reach convergence in these cases. In addition, the CPII reduces the overall computational costs by decreasing the number of iteration steps in comparison with the direct iteration approach in which intramolecular bonded interactions are excluded to ensure that induced dipole convergence is obtained. PMID- 19123852 TI - Rotational spectrum of NSF3 in the ground and v5 = 1 vibrational states: observation of Q-branch perturbation-allowed transitions with delta(k - l) = 0, +/-3, +/-6 and anomalies in the rovibrational structure of the v5 = 1 state. AB - The rotational spectrum of NSF3 in the ground and v5 = 1 vibrational states has been investigated in the centimeter- and millimeter-wave ranges. R-branch (J + 1 <-- J) transitions for J = 0, 1 and Q-branch rotational transitions for the v5 = 1 vibrational state have been measured by waveguide Fourier transform microwave spectroscopy in the range 8-26.5 GHz. The Q-branch transitions include 28 direct l-type doubling transitions (kl = +1, A1) <--> (kl = +1, A2) with J < or = 62, and 108 direct l-type resonance transitions following the selection rule delta k = delta l = +/-2 with J < or = 60 and G = |k - l| < or = 3. A process called "regional resonance" was observed in which a cluster of levels interacted strongly over a large range in J. This process led to the observation of 55 perturbation-allowed transitions following the selection rules delta(k - l) = +/ 3, +/-6. In particular, (kl = +1, A+) <--> (kl = -2, A-), (kl = +4, A+) <--> (kl = +1, A-), (kl = +2) <--> (kl = -1), (kl = +3) <--> (kl = 0), (kl = +2) <--> (kl = -3), and (kl = +3) <--> (kl = -3). The various aspects of the regional resonances are discussed in detail. An accidental near-degeneracy of the kl = 0 and kl = -4 levels at J = 26/27 led to the observation of perturbation-allowed transitions following the selection rule delta(k-l) = +/-6 with (kl = +2) <--> (kl = -4). A corresponding near-degeneracy between kl = -1 and kl = -3 levels at J = 30/31 led to the detection of similar transitions, but with (kl = +3) <--> (kl = -3). In the range 230-480 GHz, R-branch rotational transitions have been measured by absorption spectroscopy up to J = 49 in the ground-state and up to J = 50 in the v5 = 1 vibrational state. The transition frequencies have been analyzed using various reduced forms of the effective Hamiltonians. The data for the v5 = 1 vibrational state have been fitted successfully using two models up to seventh order with delta k = +/-3 interaction parameters constrained (dt constrained to zero, and epsilon to zero or to the ground-state value). On the other hand, reductions with the (delta k = +/-1, deltal = -/+2) interaction parameter q12 fixed to zero failed to reproduce the experimental data since the parameters defining the reduction transformation do not arise in the correct order of magnitude. The ground-state data have been analyzed including parameters up to fourth order constraining either parameters of the delta k = +/-3 interactions to zero (reduction A), or of the delta k = +/-6 interactions to zero (reduction B). The unitary equivalence of the different parameter sets obtained is demonstrated for both vibrational states. PMID- 19123853 TI - Theoretical study of the cyclometalated iridium(III) complexes used as chromophores for organic light-emitting diodes. AB - Time-dependent density functional theory with linear and quadratic response technology is used to calculate electronic structure, spectra, and spin-orbit coupling effects for analysis of the main mechanism for phosphorescence of the recently synthesized iridium complex [bis(2-phenylpyridine)(2-carboxy-4 dimethylaminopyridine)iridium(III)]. This compound exhibits strong green phosphorescence which is used in solution processable organic light-emitting diode devices (OLEDs) to overcome the efficiency limit imposed by the formation of triplet excitons. Attempting to foresee new structure-property relations that can guide an improved design of OLED devices based on phosphorescence of the lowest triplet state, we have conducted a theoretical analysis of the photophysical properties of a series of iridium cyclometalated complexes. PMID- 19123854 TI - Hydrogen-bonding interaction in molecular complexes and clusters of aerosol nucleation precursors. AB - Complexes and clusters bridge the gap between molecular and macroscopic levels by linking individual gaseous molecules to newly formed nanoparticles but the driving forces and mechanism for the formation of complexes and clusters in the atmosphere are not well understood. We have performed ab initio and density functional quantum chemical calculations to elucidate the role of organic acids in the formation of complexes with common atmospheric nucleating precursors such as sulfuric acid, water, and ammonia. A central feature of the complexes is the presence of two hydrogen bonds. Organic acid-sulfuric acid complexes show one strong and one medium-strength hydrogen bond whereas the corresponding hydrogen bonds in organic acid-ammonia complexes are characterized as medium-strength and weak. The formation of strong hydrogen bonds in organic acid-sulfuric acid complexes is explained by the well-established resonance-assisted hydrogen bonding theory. Organic acid-sulfuric acid and organic acid-organic acid complexes possess the largest binding energies among the homomolecular and heteromolecular dimers, about 18 kcal mol(-1) from the composite theoretical methods. Topological analysis employing quantum theory of atoms in molecules (QTAIM) shows that the charge density and the Laplacian at bond critical points (BCPs) of the hydrogen bonds of the organic acid-sulfuric acid complex (e.g., benzoic acid-sulfuric acid and cis-pinonic acid-sulfuric acid) are 0.07 and 0.16 au, respectively, which falls in or exceeds the range of one strong and one medium-strength hydrogen bonding criteria. PMID- 19123855 TI - Design of beta-hairpin peptides for modulation of cell adhesion by beta-turn constraint. AB - The CD2-CD58 interaction in immune regulation and disease pathology has provided new targets for developing potential immunosuppressive agents. In the present study, we report the introduction of constraints to generate beta-hairpin structures from the strand sequences of CD2 protein. The beta-hairpin structures were induced in the designed peptides by introducing Pro-Gly sequences in the peptides. Results from NMR and MD simulation indicated that the peptides exhibited beta-turn structure at the X-Pro-Gly-Y sequence and formed the beta hairpin structure in solution. The ability of these peptides to inhibit cell adhesion was evaluated by two cell adhesion assays. Among the peptides studied (1 4) (P1-P4), peptides 2-4 were able to inhibit cell adhesion between Jurkat cells and SRBC nearly 50% at 180 microM, and 80% inhibition between Jurkat cells and Caco-2 cells was seen at 90 microM. Peptide 1 did not show significant inhibition activity compared to control. PMID- 19123856 TI - A density functional study of the interaction of NCO with small copper clusters. AB - Density functional calculations have been carried out for Cu(n)NCO(-), Cu(n)NCO, Cu(n)NCO(+), and Cu(n)NCO(2+) clusters. It was found that for small n, charge state has a strong influence on the NCO location site. The ground states of the neutral and anionic complex clusters have a dominantly planar structure while most of the cationic complex clusters prefer three-dimensional structures. The electrostatic interaction is essential for the Cu-NCO bonding, while covalent interaction through 2pi donation strongly enhances the bonding. In neutral and anionic species the N-C bonds are strengthened and the C-O bonds are weakened, while in cationic species all the C-O distances decrease and the N-C distances can be slightly elongated in some cases, which is related to a higher NCO reactivity toward NO and O(2) to form N(2) over the positively charged Cu(n)(delta+) sites than that over the metallic Cu(n) sites. PMID- 19123857 TI - A gold(I) phosphine complex containing a naphthalimide ligand functions as a TrxR inhibiting antiproliferative agent and angiogenesis inhibitor. AB - The novel luminescent gold(I) complex [N-(N',N'-dimethylaminoethyl)-1,8 naphthalimide-4-sulfide](triethylphosphine)gold(I) was prepared and investigated for its primary biological properties. Cell culture experiments revealed strong antiproliferative effects and induction of apoptosis via mitochondrial pathways. Biodistribution studies by fluorescence microscopy and atomic absorption spectroscopy showed the uptake into cell organelles, an accumulation in the nuclei of tumor cells, and a homogeneous distribution in zebrafish embryos. In vivo monitoring of vascularisation in developing zebrafish embryos revealed a significant anti-angiogenic potency of the complex. Mechanistic experiments indicated that the inhibition of thioredoxin reductase (based on the covalent binding of a gold triethylphosphine fragment) might be involved in the pharmacodynamic behavior of this novel gold species. PMID- 19123858 TI - Electrolytically generated nanobubbles on highly orientated pyrolytic graphite surfaces. AB - Electrolysis of water is employed to produce surface nanobubbles on highly orientated pyrolytic graphite (HOPG) surfaces. Hydrogen (oxygen) nanobubbles are formed when the HOPG surface acts as a negative (positive) electrode. The coverage and volume of the nanobubbles increase with increasing voltage. The yield of hydrogen nanobubbles is much larger than the yield of oxygen nanobubbles. The growth of the individual nanobubbles during the electrolysis process is recorded in time with the help of AFM measurements and correlated with the total current. Both the size of the individual nanobubbles and the total current saturate typically after 1 min; then the nanobubbles are in a dynamic equilibrium, meaning that they do not further grow, in spite of ongoing gas production and nonzero current. The surface area of nanobubbles shows a good correlation with the nanobubble volume growth rate, suggesting that either the electrolytic gas emerges directly at the nanobubbles' surface or it emerges at the electrode's surface and then diffuses through the nanobubbles' surface. Moreover, the experiments reveal that the time constants of the current and the aspect ratio of nanobubbles are the same under all conditions. Replacement of pure water by water containing a small amount of sodium chloride (0.01 M) allows for larger currents, but qualitatively gives the same results. PMID- 19123859 TI - A Cassie-like law using triple phase boundary line fractions for faceted droplets on chemically heterogeneous surfaces. AB - We present experimental contact angle data for surfaces, which were surface engineered with a hydrophobic micropattern of hexagonal geometry. The chemically heterogeneous surface of the same hexagonal pattern of defects resulted in faceted droplets of hexagonal shape. When measuring the advancing contact angles with a viewing position aligned parallel to rows of defects, we found that an area averaged Cassie-law failed in describing the data. By replacing the area fractions by line fractions of the triple phase boundary line segments in the Cassie equation, we found excellent agreement with data. PMID- 19123860 TI - Atomic layer deposition of metal tellurides and selenides using alkylsilyl compounds of tellurium and selenium. AB - Atomic layer deposition (ALD) of metal selenide and telluride thin films has been limited because of a lack of precursors that would at the same time be safe and exhibit high reactivity as required in ALD. Yet there are many important metal selenide and telluride thin film materials whose deposition by ALD might be beneficial, for example, CuInSe2 for solar cells and Ge2Sb2Te5 for phase-change random-access memories. Especially in the latter case highly conformal deposition offered by ALD is essential for high storage density. By now, ALD of germanium antimony telluride (GST) has been attempted only using plasma-assisted processes owing to the lack of appropriate tellurium precursors. In this paper we make a breakthrough in the development of new ALD precursors for tellurium and selenium. Compounds with a general formula (R3Si)2Te and (R3Si)2Se react with various metal halides forming the corresponding metal tellurides and selenides. As an example, we show that Sb2Te3, GeTe, and GST films can be deposited by ALD using (Et3Si)2Te, SbCl3, and GeCl2 x C4H8O2 compounds as precursors. All three precursors exhibit a typical saturative ALD growth behavior and GST films prepared at 90 degrees C show excellent conformality on a high aspect-ratio trench structure. PMID- 19123861 TI - Suspensions of organic microcrystals produced in the presence of polymers: diversity of UV/Vis absorption and fluorescence properties according to the preparation conditions. AB - Free-standing microcrystals of an organic fluorescent dye, specifically, 4-n octylamino-7-nitrobenz-2-oxa-1,3-diazole, were prepared using a solvent-exchange process at room temperature, in the presence of polymers used as additives. Parameters such as the dye concentration, the nature and concentration of the polymer, and the pH of the solution were varied. Six samples of microcrystals were therefore obtained and characterized by fluorescence microscopy and by electron microscopy (TEM and SEM). They differed by their content in microcrystals, the shape and size of which depends strongly on experimental conditions. Curiously, the UV/vis absorption spectra of the microcrystal suspensions were very different from one sample to another. As a result the emission spectra were also varied. The diversity of the optical response obtained was attributed to the presence of several dye populations in the microcrystal suspensions. A distinction was made between the intrinsic spectral properties of the microcrystals and artifacts due to the presence of the additives. PMID- 19123862 TI - Protein structure refinement using 13C alpha chemical shift tensors. AB - We have obtained the (13)C alpha chemical shift tensors for each amino acid in the protein GB1. We then developed a CST force field and incorporated this into the Xplor-NIH structure determination program. GB1 structures obtained by using CST restraints had improved precision over those obtained in the absence of CST restraints and were also more accurate. When combined with isotropic chemical shifts, distance, and vector angle restraints, the root-mean squared error with respect to existing X-ray structures was better than approximately 1.0 A. These results are of broad general interest since they show that chemical shift tensors can be used in protein structure refinement, improving both structural accuracy and precision, opening up the way to accurate de novo structure determination. PMID- 19123863 TI - Eosinophilic meningitis due to Angiostrongylus and Gnathostoma species. AB - Eosinophilic meningitis can be the result of noninfectious causes and infectious agents. Among the infectious agents, Angiostrongylus cantonensis and Gnathostoma spinigerum are the most common. Although angiostrongyliasis and gnathostomiasis are not common in the United States, international travel and immigration make these diseases clinically relevant. Both A. cantonensis and G. spinigerum infection can present as severe CNS compromise. Diagnoses of both infections can be challenging and are often clinical because of a paucity of serological assays readily available in the United States. Furthermore, there are conflicting recommendations about treatment for angiostrongyliasis and gnathostomiasis. To further explore the emerging nature of these helminthic infections, a case description and review of A. cantonensis and G. spinigerum infections are presented. The clinical severity of eosinophilic meningitis and diagnosis of these infections are highlighted. PMID- 19123864 TI - Use of T cell-based diagnosis of tuberculosis infection to optimize interpretation of tuberculin skin testing for child tuberculosis contacts. AB - BACKGROUND: Treatment of recent tuberculosis infection in children aged <2 years is essential, because of high risk of progression to disease, but diagnosis is hindered by the inaccuracy of the tuberculin skin test (TST). More-accurate T cell-based tests of infection could enhance diagnosis by optimizing interpretation of the TST results. METHODS: A total of 979 child tuberculosis contacts in Istanbul underwent the TST and enzyme-linked immunospot assay. Using enzyme-linked immunospot test results as a reference standard, we assessed the effect of age and bacille Calmette-Guerin (BCG) vaccination on the sensitivity and specificity of the TST, and we computed the optimal TST cutoff points, using receiver operating characteristic curves. RESULTS: With a TST cutoff point of >or=10 mm, the sensitivity of the TST was 66% for children aged <2 years, which was lower than that for older children (P= .006). Specificity was 75% for BCG vaccinated children, compared with 92% for unvaccinated children (P= .001). Optimal cutoff points improved TST specificity for children with 1 BCG scar, with little loss of sensitivity. Despite the use of optimal cutoff points, TST sensitivity remained <70% for children aged <2 years, specificity remained <87% for BCG-vaccinated children aged >or=2 years, and overall accuracy was low for children with >1 BCG scar. CONCLUSIONS: Negative results of the TST cannot exclude tuberculosis infection for child tuberculosis contacts aged <2 years, which supports the use of preventive therapy regardless of the TST results for this age group. In children aged >or=2 years, the accuracy of the TST can be improved by adjustment of cutoff points for BCG-vaccinated children but remains poor for children with >1 BCG scar. This methodology can define optimal TST cutoff points for diagnosis of tuberculosis infection tailored to target populations. PMID- 19123867 TI - New therapies for hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection remains a global health threat with approximately 175 million carriers worldwide. Currently, treatment consists of pegylated interferon alpha plus ribavirin for 12-72 weeks, depending on HCV genotype, baseline viral load, and initial virological response to therapy. Serious adverse effects and limited sustained virological responses with this therapy warrant the need for novel HCV therapies. Specifically targeted antiviral therapies designed to inhibit the HCV serine protease and the RNA-dependent RNA polymerase have recently entered clinical development. Herein, the main characteristics of these new antiviral agents and the most important challenges arising with their use--namely, toxicities and rapid selection of resistance--are discussed. PMID- 19123865 TI - Incomplete reconstitution of T cell subsets on combination antiretroviral therapy in the AIDS Clinical Trials Group protocol 384. AB - BACKGROUND: Initiation of combination antiretroviral therapy (ART) results in higher total CD4 cell counts, a surrogate for immune reconstitution. Whether the baseline CD4 cell count affects reconstitution of immune cell subsets has not been well characterized. METHODS: Using data from 978 patients (621 with comprehensive immunological assessments) from the AIDS [Acquired Immunodeficiency Syndrome] Clinical Trials Group protocol 384, a randomized trial of initial ART, we compared reconstitution of CD4(+), CD4(+) naive and memory, CD4(+) activation, CD8(+), CD8(+) activation, B, and natural killer cells among patients in different baseline CD4(+) strata. Reference ranges for T cell populations in control patients negative for human immunodeficiency virus (HIV) infection were calculated using data from AIDS Clinical Trials Group protocol A5113. RESULTS: Patients in the lower baseline CD4(+) strata did not achieve total CD4(+) cell counts similar to those of patients in the higher strata during 144 weeks of ART, although CD4(+) cell count increases were similar. Ratios of CD4(+) naive-memory cell counts and CD4(+):CD8(+) cell counts remained significantly reduced in patients with lower baseline CD4(+) cell counts (350 cells/mm(3) achieved or approached the reference range those of control individuals without HIV infection. In contrast, patients who began ART with 0.99 for all the cases. The overall gamma values (mean +/- 2 standard deviations) were as follows: gamma = -1.74 +/- 0.47 (Model 4C) versus -1.77 +/- 0.40 (Perfexion) within 100-75% of the prescription dose; gamma = -1.57 +/- 0.26 (Model 4C) versus -1.58 +/- 0.25 (Perfexion) within 100-50% of the prescription dose; gamma = -1.47 +/- 0.18 (Model 4C) versus -1.50 +/- 0.16 (Perfexion) within 100-25% of the prescription dose. No statistical significance between the mean differences for Gamma Knife Perfexion and Model 4C was found within these ranges. The probability values were 0.65, 0.84, and 0.22, respectively. CONCLUSIONS: The use of composite sector collimators in Gamma Knife Perfexion demonstrated no statistically significant effects on the volume-averaged dose fall-off near a target periphery for typical treatment cases. PMID- 19123884 TI - Image registration accuracy of GammaPlan: a phantom study. AB - OBJECT: The authors evaluated the accuracy of the automatic image coregistration function implemented in the Leksell GammaPlan treatment planning software (Version 4C with MultiView Extension and Version 8.0). METHODS: The authors used a phantom with 9 landmarks (tips of thin cylindrical acrylic rods) evenly distributed in the treatment space. Two sets of images of the phantom were taken with both CT and MR imaging systems. The first image was obtained with the phantom aligned with the scanner's axis and the second scan was made by intentionally shifting and rotating the phantom relative to the scanner's axis. The authors attempted image registration of 2 CT image sets, CT and MR image sets, and 2 MR image sets. The accuracy of image registration was evaluated by measuring the x, y, and z coordinate values of the landmarks on each image set after 2 image sets were coregistered. The authors calculated the differences of the x, y, and z values and the distance, d, between corresponding landmarks in 2 image sets. To minimize interobserver dependence of coordinate measurements, 2 physicists did measurements independently. RESULTS: The distances, d, averaged over the 9 landmarks, were 2.63 +/- 1.64 and 0.95 +/- 0.25 mm for CT-CT and MR-MR image registrations, respectively. When the CT images of the air-filled phantom and MR images were coregistered, however, the algorithm performed poorly: d = 13.8 +/- 1.23 mm. To remedy this, the authors undertook a 2-step process by first performing landmark-based registration of the 2 image sets and subsequently applying the automatic registration. With this approach, the mean distance drastically improved: d = 0.74 +/- 0.31 mm. When the water-filled phantom was used for CT scans, the registration accuracy of CT and MR image sets was acceptable without the 2-step registration process: d = 1.18 +/- 0.36 mm. CONCLUSIONS: The accuracy of automatic registration of image sets from the same modality was within the voxel size of the scanned images. The accuracy of CT-MR image registration strongly depended on whether the phantom for CT scans was filled with air or water. This indicates the significant effect of the amount of common data available for a mutual information-based algorithm on the accuracy. PMID- 19123885 TI - Virtual framing: the feasibility of frameless radiosurgical planning for the Gamma Knife. AB - OBJECT: Gamma Knife (GK) treatments are typically delivered in 1 day with little opportunity to test different planning strategies. The authors demonstrate 2 methods for imposing GK coordinate systems upon imaging datasets without frame attachment to allow leisurely preprocedural planning, and discuss potential applications. METHODS: A "virtual framing" is constructed by coregistering a CT scan of a Leksell frame with a patient dataset using the GammaPlan (Multiview) module. Equations for skull radii are derived by approximating the skull as an ellipsoid. No proprietary software other than that of the GK system is required. In a second method, images of fiducial markers are directly superimposed on the patient dataset. Validation of the first method was achieved by comparing the lengths of 75 line segments and 60 single shot diameters measured in the virtual coordinates with those measured in real coordinates. In addition to preplanning, 2 applications are discussed. The first is the use of GK software to aid radiosurgical planning for other devices. The second is the use of virtual framing to enhance automatic optimization algorithms. RESULTS: Mean (+/- standard deviation) and root-mean-square differences in lengths were 0.18 +/- 0.32 and 0.37 mm. Mean and root-mean-square differences in diameters of single-shot plans were 0.01 +/- 0.18 and 0.18 mm. CONCLUSIONS: Virtual framing allows exploration of radiosurgical planning strategies prior to the day of treatment using only the GK software. Other applications include enhancement of radiosurgical planning for other systems and enhancement of optimization algorithms. PMID- 19123886 TI - Use of hybrid shots in planning Perfexion Gamma Knife treatments for lesions close to critical structures. AB - OBJECT: The authors investigated the use of different collimator values in different sectors (hybrid shots) when treating patients with lesions close to critical structures with the Perfexion model Gamma Knife. METHODS: Twelve patients with various tumors (6 with a pituitary tumor, 3 with vestibular schwannoma, 2 with meningioma, and 1 with metastatic lesion) that were within 4 mm of the brainstem, optic nerve, pituitary stalk, or cochlea were considered. All patients were treated at the authors' institution between June 2007 and March 2008. The patients' treatments were replanned in 2 different ways. In the first plan, hybrid shots were used such that the steepest dose gradient was aligned with the junction between the target and the critical structure(s). This was accomplished by placing low-value collimators in appropriate sectors. In the second plan, no hybrid shots were used. Sector blocking (either manual or dynamic) was required for all plans to reduce the critical structure doses to acceptable levels. Prescribed doses ranged from 12 to 30 Gy at the periphery of the target. The plans in each pair were designed to be equally conformal in terms of both target coverage (as measured by the Paddick conformity index) and critical structure sparing. RESULTS: The average number of shots required was roughly the same using either planning technique (16.7 vs 16.6 shots with and without hybrids). However, for all patients, the number of blocked sectors required to protect critical areas was larger when hybrid shots were not used. On average, nearly twice as many blocked sectors (14.8 vs 7.0) were required for the plans that did not use hybrid shots. The number of high-value collimators used in each plan was also evaluated. For small targets ( 1 cm(3)), for which 16 mm was considered a high value for the collimator, hybrid plans used an average of 1.4 times as many 16-mm sectors as did the plans without hybrids (10.7 vs 7.7 sectors). Decreasing the number of blocked sectors and increasing the number of high-value collimator sectors led to use of shorter beam-on times. Beam-on times were 1-39% higher (average 17%) when hybrid shots were not allowed. The average beam-on time for plans with and without hybrid shots was 67.4 versus 78.4 minutes. CONCLUSIONS: The judicious use of hybrid shots in patients for whom the target is close to a critical structure is an efficient way to achieve conformal treatments while minimizing the beam-on time. The reduction in beam-on time with hybrid shots is attributed to a reduced use of blocked sectors and an increased number of high-value collimator sectors. PMID- 19123887 TI - Definition of the key target volume in radiosurgical management of arteriovenous malformations: a new dynamic concept based on angiographic circulation time. AB - OBJECT: The cumulative experience worldwide indicates complete radiosurgical obliteration rates of brain arteriovenous malformations (AVMs) ranging from 35 to 90%. The purpose of this study was to propose a strategy to increase the obliteration rate for AVMs through the dynamic definition of the key target volume (KTV). METHODS: A prospective series of patients harboring an AVM was assessed using digital subtraction angiography in which a digital counter was used to measure the several stages of the frame-by-frame circulation time. All the patients were analyzed using dynamic measurement planning to define the KTV, corresponding to the volume of the shunt with the least vascular resistance and the earliest venous drainage. All patients underwent catheter-based angiography, a subgroup was additionally assessed by means of a superselective catheterization, and among these a further subgroup received embolization. The shunts were also categorized according to their angioarchitectural type: fistulous, plexiform, or mixed. The authors applied the radiosurgery-based grading system (RBGS) as well to find a correlation with the obliteration rate. RESULTS: This series includes 44 patients treated by radiosurgery; global angiography was performed for all patients, including dynamic measurement planning. Eighty-four percent of them underwent superselective catheterization, and 50% of the total population underwent embolization. In the embolized arm of the study, the pretreatment volume was up to 120 ml. In patients with a single treatment, the mean volume was 8.5 ml, and the median volume was 6.95 +/- 4.56 ml (mean +/- standard deviation), with a KTV of up to 15 ml. For prospectively staged radiosurgery, the mean KTV was 28 ml. The marginal radiation dose was 18 22 Gy, with a mean of dose 20 Gy. The mean RBGS score was 1.70. The overall obliteration rate was 91%, including the repeated radiosurgery group (4 patients), in which 100% showed complete obliteration. The overall permanent deficit was 2 of 44 patients, 1 in each group. CONCLUSIONS: Dynamic definition of the KTV might increase the obliteration rate, even in complex AVMs, allowing the treatment of smaller volumes off the recruitment vessels (pseudonidus). By using this technique, the authors avoided double-blind treatment, where the neurosurgeon does not know precisely which type of lesion he or she is irradiating and the interventionalist does not know why and what he or she is embolizing. PMID- 19123888 TI - Treatment of arteriovenous malformations using Gamma Knife surgery: the experience at the University of Washington from 2000 to 2005. AB - OBJECT: The purpose of this study was to examine the efficacy and toxicity of treating arteriovenous malformations (AVMs) with the model 3C Gamma Knife at the University of Washington Medical Center. METHODS: Ninety-five evaluable patients with 99 treatable AVMs were treated at the University of Washington Medical Center from April 2000 through June 2005. The median patient age at the time of treatment was 40 years (range 6-68 years). The male to female patient ratio was 0.98:1. The median AVM volume treated was 3.8 cm(3) (range 0.12-32 cm(3)). Forty four percent of the patients had hemorrhaged prior to treatment. The median peripheral Gamma Knife surgery dose was 20 Gy with a median of 12 isocenters treated. The median follow-up duration was 38 months (range 3-91 months). Eighty one percent of the patients had no previous stereotactic radiosurgery (SRS), whereas the remaining 19% had previously been treated with linear accelerator based SRS. RESULTS: The Kaplan-Meier estimated 6-year AVM obliteration rate for the entire cohort was 71.4%. The Kaplan-Meier estimated 6-year obliteration rate was 72% for patients having no prior SRS and 54.5% for those undergoing repeat SRS. The median time to AVM obliteration was 47 months, with 90% of the obliterations occurring between 24 and 58 months. Eight patients (7.4%) experienced late toxicities. There were 2 fatal bleeds and 13 (13.8%) nonfatal bleeds after Gamma Knife surgery. CONCLUSIONS: Gamma Knife surgery is an effective treatment for AVMs, resulting in an excellent obliteration rate with acceptable toxicity. PMID- 19123889 TI - Retrospective analysis of unstaged and staged Gamma Knife surgery with and without preceding embolization for the treatment of arteriovenous malformations. AB - OBJECT: The authors conducted a retrospective study to examine data on rates of obliteration of arteriovenous malformations (AVMs) with use of various combinations of treatment modalities based on Gamma Knife surgery (GKS). The authors believe that this study is the first to report on patients treated with embolization followed by staged GKS. METHODS: The authors identified 150 patients who underwent GKS for treatment of AVMs between 1994 and 2004. In a retrospective study, 4 independent groups emerged based on the various combinations of treatment: 92 patients who underwent unstaged GKS, 28 patients who underwent embolization followed by unstaged GKS, 23 patients who underwent staged GKS, and 7 patients who underwent embolization followed by staged GKS. A minimum of 3 years of follow-up after the last GKS treatment was required for inclusion in the retrospective analysis. Angiograms, MR images, or CT scans at follow-up were required for calculating rates of obliteration of AVMs. RESULTS: Fifty-seven of 150 patients (38%) supplied angiograms, and overall obliteration was confirmed in 43 of these 57 patients (75.4%). An additional 37 patients had follow-up MR images or CT scans. The overall obliteration rate, including patients with follow up angiograms and patients with follow-up MR images or CT scans, was 68 of 94 (72.3%). Patients who underwent unstaged GKS had a follow-up rate of 58.7% (54 of 92) and an obliteration rate of 75.9% (41 of 54). Patients who underwent embolization followed by unstaged GKS had a follow-up rate of 53.5% (15 of 28) and an obliteration rate of 60.0% (9 of 15). Patients who underwent staged GKS had a follow-up rate of 82.6% (19 of 23) and an obliteration rate of 73.7% (14 of 19). Patients who underwent embolization followed by staged GKS had a follow-up rate of 85.7% (6 of 7) and an obliteration rate of 66.7% (4 of 6). CONCLUSIONS: Gamma Knife surgery is an effective means of treating AVMs. Embolization prior to GKS may reduce AVM obliteration rates. Staged GKS is a promising method for obtaining high obliteration rates when treating larger AVMs in eloquent locations. PMID- 19123890 TI - Staged radiosurgery for extra-large cerebral arteriovenous malformations: method, implementation, and results. AB - OBJECT: The effectiveness and safety of radiosurgery for small- to medium-sized cerebral arteriovenous malformations (AVMs) have been well established. However, the management for large cerebral AVMs remains a great challenge to neurosurgeons. In the past 5 years the authors performed preplanned staged radiosurgery to treat extra-large cerebral AVMs. METHODS: An extra-large cerebral AVM is defined as one with nidus volume > 40 ml. The nidus volume of cerebral AVM is measured from the dose plan-that is, as being the volume contained within the best-fit prescription isodose. From January 2003 to December 2007, the authors treated 6 patients with extra-large AVMs by preplanned staged GKS. Staged radiosurgery is implemented by rigid transformation with translation and rotation of coordinates between 2 stages. The average radiation-targeted volume was 60 ml (range 47-72 ml). The presenting symptoms were seizure in 4 patients and a bleeding episode in 2. One patient had undergone a previous craniotomy and evacuation of hematoma. The mean interval between the 2 radiosurgical sessions was 6.9 months (range 4.5-9.1 months). The prescribed marginal dose given to the nidus volume in each stage ranged from 16 to 18.6 Gy. The expected marginal dose of total nidus was 17-19 Gy. Regular follow-up MR imaging was performed every 6 months. The mean follow-up period was 28 months (range 12-54 months). RESULTS: Most of the patients exhibited clinical improvement: relief of headache and reduced frequency of seizure attack. All patients had significant regression of nidus observed on MR imaging follow-up. Two patients had angiogram-confirmed complete obliteration of the nidus 45 and 60 months after the second-stage radiosurgical session. One patient experienced minor bleeding 8 months after the second-stage radiosurgery with mild headache. She had satisfactory recovery without clinical neurological deficit after conservative treatment. CONCLUSIONS: These preliminary results indicate that staged radiosurgery is a practical strategy to treat patients with extra-large cerebral AVMs. It takes longer to obliterate the AVMs. The observed high signal T2 changes after the radiosurgery appeared clinically insignificant in 6 patients followed up for an average of 28 months. Longer follow-up is necessary to confirm its long-term safety. PMID- 19123891 TI - Optimal timing for Gamma Knife surgery after hemorrhage from brain arteriovenous malformations. AB - OBJECT: Optimal timing of Gamma Knife surgery (GKS) after hemorrhage from brain arteriovenous malformations (AVMs) is unclear and of concern to neurosurgeons because GKS is usually performed after absorption of the hematoma. The authors investigated whether waiting for hematoma absorption is beneficial and aimed to clarify the optimal treatment timing. METHODS: The authors retrospectively studied 211 patients with AVMs who presented with hemorrhage and underwent GKS as the initial treatment. Patients were categorized into 3 groups according to the interval between the time of first hemorrhage and GKS, as follows: Group 1, 0-3 months (70 patients); Group 2, 3-6 months (62 patients); and Group 3, > 6 months (79 patients). The obliteration rates, number of hemorrhages before and after GKS, and complication rates were compared between these 3 groups. The authors also analyzed a subgroup of 127 patients who presented with intracerebral hemorrhage (ICH) to identify the influence of ICH on outcome. RESULTS: After a median follow-up of 6.3 years, the rates of obliteration, hemorrhage after treatment, and complication were not significantly different between the 3 groups even though the patients with a longer interval before GKS (Group 3) had more AVMs in eloquent areas and neurological deficits. However, the numbers of patients with preoperative hemorrhage in the interval before GKS was significantly higher in Group 3 (1, 3, and 20 patients in Group 1, 2, and 3, respectively). These results were similar in the analyses of 127 patients presenting with ICH. CONCLUSIONS: No benefit was detected in waiting for hematoma absorption until GKS after hemorrhage from AVM. Because of higher hemorrhagic risk until GKS > 6 months after hemorrhage, the authors recommend GKS within 6 months after hemorrhage. PMID- 19123892 TI - Prognostic factors and grading systems for overall survival in patients treated with radiosurgery for brain metastases: variation by primary site. AB - OBJECT: The authors conducted a study to determine whether prognostic factors and the applicability of prognostic systems vary by primary tumor site in patients treated with radiosurgery for brain metastases. METHODS: The authors evaluated data obtained in patients who underwent radiosurgery with or without whole-brain radiotherapy (WBRT) from 1991 to 2005 for newly diagnosed brain metastases. Four groups were analyzed: 1) all primary sites combined, 2) breast, 3) lung, and 4) melanoma primary sites. Kaplan-Meier, log-rank, Cox proportional hazard uni- and multivariate analysis, and recursive partitioning analysis (RPA) were used to assess prognostic factors and 4 prognostic systems: Radiation Therapy Oncology Group (RTOG) RPA, Graded Prognostic Assessment (GPA), basic score for brain metastases (BSBM), and the newly proposed Golden grading system (GGS). The GGS divides patients into 4 prognostic groups by age >or= 65 years, Karnofsky Performance Scale (KPS) score < 70, and known presence of extracranial metastases. RESULTS: Data acquired in 479 newly diagnosed patients with 1664 lesions were analyzed. The median survival time from diagnosis of brain metastases was 12.1 months; the median follow-up was 25.4 months in 73 patients who were censored. Survival and prognostic factors were equivalent for 369 patients treated with radiosurgery compared with 110 patients treated with radiosurgery and WBRT, so these subsets were combined. Multivariate analysis of all primary sites combined demonstrated age < 65 years, KPS score >or= 70, no known extracranial metastases, and or= 70 years, primary tumor control, and 3 metastases was 15.6 and 16.9 months, respectively, for breast, 16.5 and 11.3 months for lung, and 9.0 and 5.7 months for melanoma. Analysis of the 4 prognostic systems (RTOG RPA, BSBM, GPA, and GGS) showed that each prognostic system's clinical applicability varied depending on primary tumor site. The RPA confirmed that GGS and primary tumor site are significant variables for prognosis. CONCLUSIONS: Favorable prognostic factors for patients with newly diagnosed brain metastases treated with radiosurgery vary by primary site. The 4 prognostic grading systems analyzed were applicable to different primary sites depending on which prognostic factors each individual system incorporated. Therefore, the authors recommend further development and use of primary-specific prognostic systems. PMID- 19123893 TI - A validation study of a new prognostic index for patients with brain metastases: the Graded Prognostic Assessment. AB - OBJECT: The purpose of this study was to validate a new prognostic index for patients with brain metastases. This index, the Graded Prognostic Assessment (GPA), is based on an analysis of 1960 patients whose data were extracted from the Radiation Therapy Oncology Group (RTOG) database. The GPA is based on 4 criteria: age, Karnofsky Performance Scale score, number of brain metastases, and the presence/absence of extracranial metastases. Each of the 4 criteria is given a score of 0, 0.5, or 1.0, so the patient with best prognosis would have a GPA score of 4.0. METHODS: Between April 2005 and December 2006, 140 eligible patients with brain metastases were treated at the Gamma Knife Center at the University of Minnesota. The GPA score was calculated for each patient, and the score was then correlated with survival. Survival duration was calculated from the date treatment began for the brain metastases. Eligibility criteria included patients treated with whole-brain radiation therapy, stereotactic radiosurgery, or both. RESULTS: The median survival time in months observed in the RTOG and Minnesota data by GPA score was as follows: GPA 3.5-4.0, 11.0 and 21.7; GPA 3.0, 8.9 and 17.5; GPA 1.5-2.5, 3.8 and 5.9; and GPA 0-1.0, 2.6 and 3.0, respectively. CONCLUSIONS: The University of Minnesota data correlate well with the RTOG data and validate the use of the GPA as an effective prognostic index for patients with brain metastases. Clearly, not all patients with brain metastases have the same prognosis, and treatment decisions should be individualized accordingly. The GPA score does appear to be as prognostic as the RPA and is less subjective (because the RPA requires assessment of whether the primary disease is controlled), more quantitative, and easier to use and remember. A multi institutional validation study of the GPA is ongoing. PMID- 19123894 TI - Neuroimaging and quality-of-life outcomes in patients with brain metastasis and peritumoral edema who undergo Gamma Knife surgery. AB - OBJECT: Gamma Knife surgery (GKS) has been shown to be effective for treating many patients with brain metastasis. Some brain metastases demonstrate significant peritumoral edema; radiation may induce cerebral edema or worsening preexisting edema. This study was conducted to evaluate the imaging and neurobehavioral outcomes in patients with preexisting peritumoral edema who then undergo GKS. METHODS: Between August 2003 and January 2008, 63 cases of brain metastasis with significant peritumoral edema (> 20 cm(3)) were prospectively studied. The study inclusion criteria were as follows: 1) a single metastatic lesion with significant edema (perilesional edema signal volume on FLAIR > 20 cm(3)); and 2) inclusion of only 1 lesion > 20 cm(3) in the study (in cases of multiple lesions noted on FLAIR images). All patients received MR imaging with pulse sequences including T1-weighted imaging and FLAIR with or without contrast and T2-weighted imaging at an interval of 3 months. A neurological assessment and Brain Cancer Module (BCM-20) questionnaire were obtained every 2-3 months. Kaplan Meier, Cox regression, and logistic regression were used for analysis of survival and associated factors. RESULTS: At the time of GKS, the median Karnofsky Performance Scale (KPS) score was 70 (range 50-90), and the mean BCM-20 score was 45.5 +/- 6.1. The mean tumor volume (+/- standard deviation) was 5.2 +/- 4.6 cm(3) with corresponding T2-weighted imaging and FLAIR volumes of 59.25 +/- 37.3 and 62.1 +/- 38.8 cm(3), respectively (R(2) = 0.977, p < 0.001). The mean edema index (volume of peritumoral edema/tumor volume) was 17.5 +/- 14.5. The mean peripheral and maximum GKS doses were 17.4 +/- 2.3 and 35 +/- 4.7 Gy, respectively. The median survival was 11 months. The longer survival was related to KPS scores >or= 70 (p = 0.008), age < 65 years (p = 0.022), and a reduction of > 6 in BCM-20 score (p = 0.007), but survival was not related to preexisting edema or tumor volume. A reduction in BCM-20 score of > 6 was related to decreased volume in T1-weighted and FLAIR imaging (p < 0.001). Thirty-eight (79.2%) of 48 patients demonstrated decreased tumor volume and accompanied by decreased T2-weighted imaging and FLAIR volume. Eight (16.7%) of the 48 patients exhibited increased or stable tumor volume. A margin dose > 18 Gy was more likely to afford tumor reduction and resolution of peritumoral edema (p = 0.005 and p = 0.006, respectively). However, prior external-beam radiation therapy correlated with worsened preexisting peritumoral edema (p = 0.013) and longer maintenance of corticosteroids (p < 0.001). CONCLUSIONS: Patients demonstrating a reduction in the BCM-20 score > 6, age < 65 years, and KPS score >or= 70 exhibited longer survival. Significant preexisting edema did not influence the tumor response or clinical outcome. The resolution of edema was related to better quality of life but not to longer survival. PMID- 19123895 TI - Gamma Knife surgery for the treatment of melanoma metastases: the effect of intratumoral hemorrhage on survival. AB - OBJECT: Gamma Knife surgery (GKS) improves overall survival in patients with malignant melanoma metastatic to the brain. In this study the authors investigated which patient- or treatment-specific factors influence survival of patients with melanoma brain metastases; they pay particular interest to pre- and post-GKS hemorrhage. METHODS: Demographic, treatment, and survival data on 59 patients with a total of 208 intracranial metastases who underwent GKS between 1998 and 2007 were abstracted from treatment records and from the Connecticut Tumor Registry. Multivariate analysis was used to identify factors that independently affected survival. RESULTS: Survival was significantly better in patients with solitary metastasis (p = 0.04), lesions without evidence of pre-GKS hemorrhage (p = 0.004), and in patients with total tumor volume treated < 4 cm(3) (p = 0.02). Intratumoral bleeding occurred in 23.7% of patients pre-GKS. Intratumoral bleeding occurred at a mean of 1.8 months post-GKS at a rate of 15.2%. Unlike the marked effect of pretreatment bleeding, posttreatment bleeding did not independently affect survival. Sex, systemic control, race, metastases location, whole-brain radiation therapy, chemotherapy, history of antithrombotic medications, and cranial surgery had no independent association with survival. CONCLUSIONS: These data corroborate previous findings that tumor burden (either as increased number or total volume of lesions) at the time of GKS is associated with diminished patient survival in those with intracerebral melanoma metastases. Patients who were noted to have hemorrhagic melanoma metastases prior to GKS appear to have a worse prognosis following GKS compared with patients with nonhemorrhagic metastases, despite similar rates of bleeding pre- and post-GKS treatment. Gamma Knife surgery itself does not appear to increase the rate of hemorrhage. PMID- 19123896 TI - Prospective trial of gross-total resection with Gliadel wafers followed by early postoperative Gamma Knife radiosurgery and conformal fractionated radiotherapy as the initial treatment for patients with radiographically suspected, newly diagnosed glioblastoma multiforme. AB - OBJECT: The purpose of this study was to determine whether increased local control and improved survival can be achieved in patients with glioblastoma multiformes (GBMs) who undergo aggressive resection, Gliadel wafer implantation, Gamma Knife radiosurgery (GKS), and fractionated radiotherapy (RT) as the initial treatment. METHODS: Thirty patients with radiographically suspected GBMs were screened for enrollment in a Phase I/II prospective clinical trial. Twenty-seven patients were eligible and underwent gross-total resection and Gliadel wafer implantation. Gamma Knife radiosurgery (12 Gy at 50%) was administered to the resection cavity within 2 weeks of surgery. Patients then received standard fractionated RT (total dose 60 Gy over 6 weeks). Temozolomide was prescribed for patients at the time of recurrence. Surveillance MR imaging, neurological examination, and quality-of-life evaluations were performed at 2-month intervals. To estimate the potential effects on the DNA repair mechanism, tumor tissue was analyzed with methylation-specific polymerase chain reaction analysis and immunohistochemical assays for MGMT gene promoter methylation and protein expression. RESULTS: The median survival for all patients was 50 weeks and the 2 year survival rate was 22%. When stratified into standard and high-risk patient groups, the median survivals were 76 and 33 weeks, respectively. Two patients remain alive at the time of this report with no clinical or radiographic evidence of disease at > 189 and 239 weeks posttreatment and excellent performance status. Local tumor control was achieved in 53% of patients, and local failure occurred in 47%. No acute early toxicity was noted; however, delayed symptomatic radionecrosis occurred in 47% of patients, which required repeated operations 9 24 months after the initial treatment. Delayed hydrocephalus requiring ventriculoperitoneal shunt placement occurred in 47% of patients. There was a significant difference in survival between patients whose tumors contained the methylated and unmethylated MGMT promoter, 103 versus 45 weeks, respectively (p = 0.0009, log-rank test). CONCLUSIONS: The combination of aggressive resection, Gliadel wafer implantation, and GKS in addition to standard fractionated RT in selected patients resulted in increased local control and increased survival compared with a historical control group treated with surgery and involved-field RT alone. Delayed focal radionecrosis was increased to 47% in this series and was managed with steroids and repeated resection. Aggressive local tumor control with these multimodal therapies should be approached judiciously for a select group of high performance patients and the probability of developing symptomatic radionecrosis requiring surgery should be anticipated and fully disclosed to patients who undergo this treatment. PMID- 19123897 TI - Gamma Knife surgery for metastatic brain tumors. AB - OBJECT: The authors compared results of Gamma Knife surgery (GKS) for brain metastases obtained at 2 institutions in Japan. METHODS: They analyzed a consecutive series of 2390 patients with brain metastases who underwent GKS from 1998 through 2005 in 2 institutes (1,181 patients in Chiba; 1,209 in Mito). In the 2 facilities, 1 neurosurgeon each was responsible for diagnosis, patient selection, GKS procedures, and follow-up (T.S. in Chiba, M.Y. in Mito). Even if tumor numbers exceeded 4, all visible lesions were irradiated with a total skull integral dose (TSID) of or= 90% coverage versus 0.0% for lesions receiving < 90% (p = 0.00048). Median overall survival was 5.1 months after GKS and 8.4 months after the discovery of brain metastases. Univariate analysis revealed that KPS score (p = 0.000004), recursive partitioning analysis class (p = 0.00043), and single metastases (p = 0.028), but not more than 3 metastases, to be prognostic factors of overall survival. The KPS score remained significant after multivariate analysis. Overall survival for patients with a KPS score >or= 70 was 7.1 months compared with 1.3 months for a KPS score 3 metastases. Higher rates of local tumor control were achieved for RCC in comparison with melanoma, and this may have an effect on survival in some patients. Although outcomes generally remained poor in this study population, these results suggest that GKS can be considered as a treatment option for many patients with radioresistant brain metastases, even if these patients have multiple lesions. PMID- 19123899 TI - Hearing preservation in vestibular schwannoma stereotactic radiosurgery: what really matters? AB - Stereotactic radiosurgery (SRS) for vestibular schwannomas has evolved and improved over time. Although early short-term follow-up reports suggest that fractionation yields hearing preservation rates equivalent to modern single-dose SRS techniques, significant questions remain regarding long-term tumor control after the use of fractionation in a late responding tumor with a low proliferative index and alpha/beta ratio. With single-dose SRS, critical hearing preservation variables include: 1) strict attention to prescription dose 3D conformality so that the ventral cochlear nucleus (VCN) receives or= 60-month follow-up available in 27 of these patients, >or= 48 months in 32, >or= 36 months in 38, >or= 24 months in 43, >or= 12 months in 51, and >or= 6 months in 54 (1 patient was excluded from the analysis because the follow-up was < 6 months). Various patterns of changes in vestibular function were observed in either positive or negative directions. A significant difference in total DHI score was seen only in the elderly (> 65 years old) patients pre-GKS compared with post-GKS (t = 1.34, p = 0.05). CONCLUSIONS: Longitudinal changes in vestibular function occur over time, with the largest changes seen in the first 6 months after treatment. Potential for clinical intervention, such as vestibular rehabilitation therapy, exists during this interval; however, larger cohorts must be studied to determine the timing and efficacy of this intervention. The statistically significant improvement in the DHI score in the patient cohort > 65 years old treated with GKS suggests that this group may benefit from this option when considering the symptom of dizziness. PMID- 19123901 TI - Retreatment of vestibular schwannomas with Gamma Knife surgery. AB - OBJECT: The response rate of vestibular schwannomas (VSs) to radiosurgery has reached the 97% level in several published series. When failure rarely occurs, some controversy has existed as to whether the tumor has to be resected or can be safely retreated with radiosurgery. The authors retrospectively studied the outcome of retreating 11 patients with Gamma Knife surgery (GKS). METHODS: The authors studied 11 patients at the New England Gamma Knife Center who had undergone GKS as a second radiosurgical treatment for VS from 1994 to 2007. One patient underwent proton-beam radiotherapy as the first treatment, and the other 10 patients had undergone GKS initially. Tumor control (size before and after the first and the second treatment) was evaluated using MR imaging to demonstrate the course after the 2 treatments. Facial nerve function (House-Brackmann grading system), trigeminal nerve function, hearing (Gardner-Robertson classification), and any adverse radiation effects were evaluated. The prescription dose was 12 Gy (11-13.2 Gy) for both treatments. RESULTS: Of the 11 patients, 2 showed increase, 1 had no change, and, in 8, the VS decreased in size after the retreatment. One tumor remained unchanged over the first 6 months, but demonstrated signs of internal necrosis. All patients demonstrated stable facial nerve function. Regarding facial numbness, 2 patients experienced increases, 8 no change, and 1 decreased numbness. There was no functional hearing prior to the second treatment in 10 patients, and hearing was impaired in 1 patient. Adverse radiation effects (slight peduncular edema) were seen in 2 patients after the second treatment, and 1 patient had edema after the first treatment as well. CONCLUSIONS: Vestibular schwannomas can be retreated with GKS with good tumor control response and low risk of toxicity. PMID- 19123902 TI - Gamma Knife surgery for schwannomas originating from cranial nerves III, IV, and VI. AB - OBJECT: Schwannomas from the motor cranial nerves controlling eye movement are rare. The authors evaluated the role of Gamma Knife surgery (GKS) in the management of schwannomas originating from cranial nerves III, IV, and VI. METHODS: Over a 7-year period, 8 patients with schwannomas originating from the oculomotor (2 patients), trochlear (5 patients), or abducent (1) nerve underwent GKS. The mean patient age was 46.1 years (range 19-59 years). The presenting symptoms included diplopia in 5 patients, ptosis in 1 patient, ophthalmoplegia in 1 patient, and headache in 1 patient. Two patients had a history of neurofibromatosis Type 2. Gamma Knife surgery was performed as primary management in 7 patients and after prior resection in 1 patient. The median and mean tumor volumes were 0.22 and 1.32 cm(3) (range 0.03-7.4 cm(3)). A median margin dose of 12.5 Gy (range 11.0-13.0 Gy) was prescribed to the tumor margin. Clinical and imaging follow-up data were available for all 8 patients. RESULTS: Magnetic resonance imaging showed tumor regression in all patients. The progression-free period varied from 4 to 42 months, with a mean of 21 months. Over a mean of 23 months, 4 of the 5 patients with a trochlear schwannoma and symptoms of diplopia noted symptomatic improvement. No improvement was noted in the 2 patients with oculomotor nerve palsies. Headache was improved in the 1 patient with an abducent neuroma. CONCLUSIONS: Gamma Knife surgery is an effective and minimally invasive approach capable of inactivating schwannomas originating from the oculomotor, trochlear, and abducent nerves. Accompanying trochlear function may improve. Longer follow-up and larger patient samples are needed to confirm the authors' initial observations. PMID- 19123903 TI - Gamma Knife surgery for trigeminal pain caused by benign brain tumors. AB - OBJECT: The authors report the effects of Gamma Knife surgery (GKS) on benign tumor-related trigeminal pain in patients who underwent follow-up for a mean 57.8 months. METHODS: From 1999 to 2004, 21 patients with benign tumor-related trigeminal pain (12 meningiomas and 9 schwannomas) underwent GKS as a primary or repeated treatment. These patients harbored tumors within the radiosurgical target area. For meningiomas, the mean radiosurgical treatment volume was 8.2 ml (range 1.1-21 ml), and the mean radiosurgical tumor margin dose was 12.7 Gy (range 12-15 Gy); for schwannomas, the mean volume was 5.6 ml (range 2-9.2 ml), and the mean marginal dose was 13 Gy (range 11.5-16 Gy). Seven patients underwent retreatment for recurrent or persistent pain; the ipsilateral trigeminal nerve or ganglion was identified and a mean maximal dose of 60.7 Gy (range 40-70 Gy) was delivered to these targets. In 1 patient undergoing retreatment, the margin dose was 12 Gy. The mean age at the time of radiosurgery was 54.5 years (range 18-79 years). RESULTS: The mean follow-up period was 57.8 months (range 36-94 months). Overall, 12 (57%) of 21 patients experienced pain relief without medication after the first GKS and the mean time to drug discontinuation was 10.5 months (range 2 24 months). Initial pain improvement was noted in 17 patients (81%) with a mean time of 3.7 months (range 1 week-10 months) after GKS. Eight patients underwent repeated GKS for persistent and recurrent pain. Four patients (50%) had complete pain relief. The final results of the first and repeated GKS were excellent in 16 patients (76%), and in only 1 patient did GKS fail, and this patient later underwent open surgery. For all 21 patients (100%), control of tumor growth was documented at a mean of 46 months after GKS. Three of 6 patients with pre-GKS facial numbness reported improvement, but 4 suffered new facial numbness after repeated GKS. CONCLUSIONS: Gamma Knife surgery appears to be an effective tool to treat benign tumor-related trigeminal pain and control tumor growth. Repeated GKS targeting the trigeminal root or ganglion can be considered a tool to enhance the efficacy of pain management if pain persists or recurs, but the optimum treatment dose needs further investigation. PMID- 19123904 TI - A prospective cohort study of microvascular decompression and Gamma Knife surgery in patients with trigeminal neuralgia. AB - OBJECT: The aim of this study was to analyze 1 surgeon's 4-year experience with microvascular decompression ([MVD], 36 patients) and Gamma Knife surgery ([GKS], 44 patients) in 80 consecutive patients with trigeminal neuralgia (TN). METHODS: The authors conducted a prospective cohort study from March 1999 to December 2003 with an independent clinical assessment of the results and serial patient satisfaction surveys. All patients completed a 2004 patient satisfaction survey (0.5-5 years postoperative), and 70% of surviving patients completed the same survey in 2007 (3.9-8.5 years postoperative). Follow-up was undertaken in 100% of the patients (mean 3.4 +/- 2.14 years, range 0.17-8.5 years). RESULTS: Respective initial and latest follow-up raw pain-free rates were 100 and 80.6% for MVD and 77.3 and 45.5% for GKS. The median time to the maximal benefit after GKS was 4 weeks (range 1 week-6 months). Respective initial, 2-, and 5-year actuarial pain free rates were 100, 88, and 80% for MVD and 78, 50, and 33% for GKS (p = 0.0002). The relative risk of losing a pain-free status by 5 years posttreatment was 3.35 for patients in the GKS group compared with the MVD group. Initial and 5 year actuarial rates for >or= 50% pain relief after GKS were 100 and 80%, respectively. The respective rates of permanent mild and severe sensory loss were 5.6 and 0% for patients in the MVD group, as opposed to 6.8 and 2.3% for patients in the GKS group. Anesthesia dolorosa did not occur during the study. Both procedures enjoyed a high degree of early patient satisfaction (95-100%). Microvascular decompression maintained the same rate of patient satisfaction, but satisfaction with GKS decreased to 75% as pain control waned. Twenty-three patients (29%) died of causes unrelated to the TN or the surgical intervention during the follow-up, and their pain status was known at the time of death. Statistically significant intergroup differences for the MVD versus GKS cohorts were age (median 54 years, range 36-70 years vs median 74 years, range 48-92 years, respectively), preoperative symptom duration (median 2.58 years, range 0.33-15 years vs median 7.5, range 0.6-40 years, respectively), and the presence of major comorbidities (2.8 vs 58.3%, respectively). CONCLUSIONS: In this nonrandomized prospective cohort trial of selected patients with potentially relevant intergroup differences, MVD was significantly superior to GKS in achieving and maintaining a pain-free status in those with TN and provided similar early and superior longer-term patient satisfaction rates compared with those for GKS. The complications of wound cerebrospinal fluid leakage, hearing loss, and persistent diplopia (1 case each in the MVD group) were not seen after GKS. PMID- 19123905 TI - Clinical outcomes after Gamma Knife surgery for idiopathic trigeminal neuralgia: review of 76 consecutive cases. AB - OBJECT: Stereotactic radiosurgery is an increasingly used, and the least invasive, surgical option for patients with trigeminal neuralgia (TN). In this study, the authors performed a retrospective evaluation of the safety and efficacy of this method for idiopathic TN. METHODS: The authors reviewed data from 76 patients with idiopathic TN who underwent Gamma Knife surgery (GKS). The mean age of the patients was 64 years (range 27-83 years). All patients had typical features of TN. Thirty patients (39.5%) had previously undergone surgery. The intervention consisted of GKS on the retrogasserian cisternal portion of the fifth cranial nerve. The mean maximum GKS dose used was 85.1 Gy (range 75-90 Gy). RESULTS: Patients were followed-up from 6 to 42 months (mean 20.3 months) after GKS. Complete pain relief was achieved in 83.1% of the patients within 1 year, 70.9% within 2 years, and 62.5% within 3 years. Patients who underwent previous surgery demonstrated a lower rate of pain relief (p < 0.05). Twenty patients (26.3%) reported pain recurrence between 6 and 42 months after treatment. New or worsened persistent trigeminal dysfunction developed after GKS in 16 patients (21%); 8 of these patients described some facial numbness/not bothersome, and 8 reported some facial numbness/somewhat bothersome. None of the patients developed troublesome dysesthesia or anesthesia dolorosa. CONCLUSIONS: Gamma Knife surgery for idiopathic TN proved to be safe and effective and was associated with a particularly low rate of complications. PMID- 19123906 TI - Gamma Knife surgery used as primary and repeated treatment for idiopathic trigeminal neuralgia. AB - OBJECT: The purpose of this study was to assess the outcome of idiopathic trigeminal neuralgia (TN) treated with Gamma Knife surgery (GKS) as a primary and repeated treatment modality with a mean follow-up of 5.7 years. METHODS: Between July 1999 and September 2005, a total of 89 patients with idiopathic TN underwent GKS as a primary treatment. The entry zone of the TN was targeted with a 4-mm collimator and treated with a maximal dose of 60-90 Gy (mean 79 Gy). The dose to the pontine margin was always kept < 15 Gy. Twenty patients received repeated GKS for recurrent or residual pain with a maximal dose of 40-76 Gy (mean 52 Gy). For the second procedure, the target was positioned at the same location as the first treatment. RESULTS: The mean follow-up period was 68 months (range 32-104 months). Sixty-nine (77.5%) of the 89 patients experienced a favorable response, as follows: 50 (56%) had excellent, 12 (13.5%) had good, and 7 (7.8%) had fair outcomes. The mean time to pain relief was 1.1 months (range 2 days-6 months). No significant correlation, but more likely a tendency, was found between the dose and pain relief (p = 0.08). Also, no correlation was noted for facial numbness (p = 0.77). The mean follow-up period after repeated GKS was 60 months (range 32-87 months). Outcomes after repeated GKS were excellent in 11 patients (55%) and good in 1 (5%). Seven patients experienced facial numbness. No correlation was found between the additive dose and pain relief (p = 0.24) or facial numbness (p = 0.15). Final outcomes of primary and repeated GKS were excellent in 61 (68.5%), good in 13 (14.6%), and fair in 7 (7.9%). In total, 91% of the patients were successfully treated with this method. There was no statistical significance for efficacy between primary and repeated GKS (p = 0.65), but there was a significant difference for facial numbness (p = 0.007). CONCLUSIONS: Gamma Knife surgery established durable pain relief when used as a primary and repeated surgery. Treatment was successful for a total of 91% of patients at a mean follow-up of 5.7 years, but facial numbness was also relatively higher. PMID- 19123907 TI - Comparison of the results of 2 targeting methods in Gamma Knife surgery for trigeminal neuralgia. AB - OBJECT: Gamma Knife surgery (GKS) is an effective treatment for intractable trigeminal neuralgia (TN). The authors compared results using two major GKS target points, the dorsal root entry zone and the retrogasserian portion, in a series of patients with intractable TN. METHODS: One hundred patients with medically refractory TN underwent GKS between August 1998 and December 2007. Thirty-seven were men, and 63 were women. The median age at GKS was 74 years. With a single isocenter and use of a 4mm collimator, 51 patients received 80 Gy at the proximal trigeminal nerve (posterior group) and 7 patients received 80 Gy, 1 patient received 85 Gy, and 41 patients received 90 Gy at the retrogasserian portion (anterior group). Follow-up was obtained by clinic visits every 3-6 months after GKS. Data on pain control, complications, and pain recurrence were recorded. The relationships between pain control status, complications, recurrence, and the target portions (anterior vs posterior) were analyzed. RESULTS: The median duration of follow-up was 30 months (range 3-88 months). Initially, 87 patients achieved pain-free status and 64 achieved complete remission. At the final follow-up visit, 68 patients were still in pain-free status and 42 were in complete remission. Recurrence of facial pain occurred in 15 patients. Forty-one patients developed some degree of trigeminal dysfunction. The rate of initial complete remission was higher in the posterior group than in the anterior group (p = 0.003). More complications were observed in the anterior group than in the posterior group (p = 0.009). CONCLUSIONS: The posterior targeting group had better pain control and a lower complication rate. The authors recommend the posterior targeting method and use of 80 Gy for treatment of TN with GKS. PMID- 19123908 TI - Characterization and adsorption modeling of silicon carbide-derived carbons. AB - We present characterization results of silicon carbide-derived carbons (Si-CDCs) prepared from both nano- and micron-sized betaSiC particles by oxidation in pure chlorine atmosphere at various synthesis temperatures (600-1000 degrees C). Subsequently, the adsorption modeling study of simple gases (CH4 and CO2) in these Si-CDC samples for a wide range of pressures and temperatures using our Finite Wall Thickness model [Nguyen, T. X.; Bhatia, S. K. Langmuir 2004, 20, 3532] was also carried out. In general, characterization results showed that the core of Si-CDC particles contains predominantly amorphous material while minor graphitization was also observed on the surface of these particles for all the investigated synthesis temperatures (600-1000 degrees C). Furthermore, postsynthetic heat treatment at 1000 degrees C for 3 days, as well as particle size of precursor (betaSiC) were shown to have slight impact on the graphitization. In spite of the highly disordered nature of Si-CDC samples, the adsorption modeling results revealed that the Finite Wall Thickness model provides reasonably good prediction of experimental adsorption data of CO2 and CH4 in all the investigated Si-CDC samples at the temperatures of 273 K, 313 K, and 333 K for a wide range of pressure up to 200 bar. Furthermore, the impact of the difference in molecular size and geometry between analysis and probing gases on the prediction of the experimental adsorption isotherm in a disordered carbon using the slit-pore model is also found. Finally, the correlation between compressibility of the Si-CDC samples under high pressure adsorption and their synthesis temperature was deduced from the adsorption modeling. PMID- 19123909 TI - Direct-write patterning palladium colloids as a catalyst for electroless metallization for microwave composites. AB - Patterning of metal colloids by inkjet printing on paper is demonstrated as a precursor to electroless metallization. The development of the metal pattern is followed in terms of the conductivity and mass of the metal deposited and is shown to have critical phase behavior. The utility of this technique for large area microscale patterning is demonstrated. Sample patterns of frequency selective surface designs were manufactured and shown to conform to computationally modeled expectations in the microwave regime. PMID- 19123911 TI - Importance of van der Waals interactions in liquid water. AB - We present ab initio molecular dynamics studies on liquid water using density functional theory in conjunction with either dispersion-corrected atom-centered potentials or empirical van der Waals corrections. Our results show that improving the description of van der Waals interactions in DFT-GGA leads to a softening of liquid water's structure with higher mobility. The results obtained with dispersion-corrected atom-centered potentials are especially encouraging. In particular, the radial distribution functions are in better agreement with experiment, and the self-diffusion coefficient increases by more than three-fold compared with the one predicted by the BLYP functional. This work demonstrates that van der Waals interactions are essential in fine-tuning both structural and dynamical properties of liquid water. PMID- 19123912 TI - Investigations of strong hydrogen bonding in (ROH)n...FHF- (n = 1, 2 and R = H, CH3, C2H5) clusters via high-pressure mass spectrometry and quantum calculations. AB - An examination of strong hydrogen bonds found in (ROH)(n)...FHF(-) clusters (n = 1 and 2; R = H, CH(3), C(2)H(5)) is presented. Excellent agreement is observed between thermochemical values obtained from high-pressure mass spectrometric measurements and those predicted from MP2(full)/6-311++G(d,p)//B3LYP/6 311++G(d,p) calculations. Calculated structures are examined, and insight into the geometric nature of the bonding for these systems is obtained. In the case of water binding to FHF(-), it was found that the large entropic advantage of one particular structure, which was not the most enthalpically favored, was significant enough to make it the predominant species within the ion source. In the case of methanol solvation, no evidence of secondary interaction of the methyl group and any other moiety could be found. The structural details revealed from calculations of the ethanol-solvated clusters indicate that secondary interactions between the terminal methyl group and FHF(-) have an impact on the length of the FHF and OHF bonds. PMID- 19123914 TI - Methyl group dynamics in polycrystalline and liquid ubiquinone Q(0) studied by neutron scattering. AB - We present a quasi-elastic neutron scattering (QENS) study on the methyl group dynamics of ubiquinone Q(0) in the solid and liquid state. For solid ubiquinone Q(0), the dynamics can be described with three Lorentzian functions in the framework of a jump model among three equidistant sites on a circle. According to the known molecular structure of Q(0) in the solid state, this is consistent with three nonequivalent methyl groups in the molecule. From the temperature-dependent analysis of the QENS spectra, the activation energies were determined. The barrier heights could be evaluated from librational bands in the inelastic part of the spectra. The results from neutron spectroscopy are compared to Gaussian 03 calculations leading to an assignment of the activation energies to the different methyl groups in Q(0). The dynamics of Q(0) in the liquid state is evaluated with a scattering function taking into account three different molecular motions. It is demonstrated that the temperature dependence of the long-range diffusion and isotropic rotational diffusion exhibit an Arrhenius-like behavior, whereas the process of methyl group rotation in the liquid phase is virtually free of a barrier. PMID- 19123915 TI - Cyclic versus linear isomers produced by reaction of the methylidyne radical (CH) with small unsaturated hydrocarbons. AB - The reactions of the methylidyne radical (CH) with ethylene, acetylene, allene, and methylacetylene are studied at room temperature using tunable vacuum ultraviolet (VUV) photoionization and time-resolved mass spectrometry. The CH radicals are prepared by 248 nm multiphoton photolysis of CHBr(3) at 298 K and react with the selected hydrocarbon in a helium gas flow. Analysis of photoionization efficiency versus VUV photon wavelength permits isomer-specific detection of the reaction products and allows estimation of the reaction product branching ratios. The reactions proceed by either CH insertion or addition followed by H atom elimination from the intermediate adduct. In the CH + C(2)H(4) reaction the C(3)H(5) intermediate decays by H atom loss to yield 70(+/-8)% allene, 30(+/-8)% methylacetylene, and less than 10% cyclopropene, in agreement with previous RRKM results. In the CH + acetylene reaction, detection of mainly the cyclic C(3)H(2) isomer is contrary to a previous RRKM calculations that predicted linear triplet propargylene to be 90% of the total H-atom coproducts. High-level CBS-APNO quantum calculations and RRKM calculations for the CH + C(2)H(2) reaction presented in this manuscript predict a higher contribution of the cyclic C(3)H(2) (27.0%) versus triplet propargylene (63.5%) than earlier predictions. Extensive calculations on the C(3)H(3) and C(3)H(2)D system combined with experimental isotope ratios for the CD + C(2)H(2) reaction indicate that H atom-assisted isomerization in the present experiments is responsible for the remaining discrepancy between the new RRKM calculations and the experimental results. Cyclic isomers are also found to represent 30(+/-6)% of the detected products in the case of CH + methylacetylene, together with 33(+/-6)% 1,2,3 butatriene and 37(+/-6)% vinylacetylene. The CH + allene reaction gives 23(+/-5)% 1,2,3-butatriene and 77(+/-5)% vinylacetylene, whereas cyclic isomers are produced below the detection limit in this reaction. The reaction exit channels deduced by comparing the product distributions for the aforementioned reactions are discussed in detail. PMID- 19123916 TI - Thrombospondin-2 influences the proportion of cartilage and bone during fracture healing. AB - Thrombospondin-2 (TSP2) is a matricellular protein with increased expression during growth and regeneration. TSP2-null mice show accelerated dermal wound healing and enhanced bone formation. We hypothesized that bone regeneration would be enhanced in the absence of TSP2. Closed, semistabilized transverse fractures were created in the tibias of wildtype (WT) and TSP2-null mice. The fractures were examined 5, 10, and 20 days after fracture using microCT, histology, immunohistochemistry, quantitative RT-PCR, and torsional mechanical testing. Ten days after fracture, TSP2-null mice showed 30% more bone by microCT and 40% less cartilage by histology. Twenty days after fracture, TSP2-null mice showed reduced bone volume fraction and BMD. Mice were examined 5 days after fracture during the stage of neovascularization and mesenchymal cell influx to determine a cellular explanation for the phenotype. TSP2-null mice showed increased cell proliferation with no difference in apoptosis in the highly cellular fracture callus. Although mature bone and cartilage is minimal 5 days after fracture, TSP2-null mice had reduced expression of collagen IIa and Sox9 (chondrocyte differentiation markers) but increased expression of osteocalcin and osterix (osteoblast differentiation markers). Importantly, TSP2-null mice had a 2-fold increase in vessel density that corresponded with a reduction in vascular endothelial growth factor (VEGF) and Glut-1 (markers of hypoxia inducible factor [HIF]-regulated transcription). Finally, by expressing TSP2 using adenovirus starting 3 days after fracture, chondrogenesis was restored in TSP2-null mice. We hypothesize that TSP2 expressed by cells in the fracture mesenchyme regulates callus vascularization. The increase in vascularity increases tissue oxemia and decreases HIF; thus, undifferentiated cells in the callus develop into osteoblasts rather than chondrocytes. This leads to an alternative strategy for achieving fracture healing with reduced endochondral ossification and enhanced appositional bone formation. Controlling the ratio of cartilage to bone during fracture healing has important implications for expediting healing or promoting regeneration in nonunions. PMID- 19123917 TI - Dispersal in a statistically structured population: fat tails revisited. AB - Dispersal has long been recognized as a crucial factor affecting population dynamics. Several studies on long-distance dispersal revealed a peculiarity now widely known as a problem of "fat tail": instead of the rate of decay in the population density over large distances being described by a normal distribution, which is apparently predicted by the standard diffusion approach, field data often show much lower rates such as exponential or power law. The question as to what are the processes and mechanisms resulting in the fat tail is still largely open. In this note, by introducing the concept of a statistically structured population, we show that a fat-tailed long-distance dispersal is a consequence of the fundamental observation that individuals of the same species are not identical. Fat-tailed dispersal thus appears to be an inherent property of any real population. We show that our theoretical predictions are in good agreement with available data. PMID- 19123920 TI - Stacking of bacteriochlorophyll c macrocycles in chlorosome from Chlorobium limicola as revealed by intermolecular 13C magnetic-dipole correlation, X-ray diffraction, and quadrupole coupling in 25Mg NMR. AB - The stacking of the bacteriochlorophyll (BChl) c macrocycles and the role of water in forming an aggregate sheet, in chlorosome, were examined by means of (13)C NMR spectroscopy, the measurement of the X-ray diffraction pattern, and (25)Mg NMR spectroscopy. (1) The stacking of the macrocycles, i.e., weakly overlapped dimers forming displaced layers, was selected out of six different kinds of stacking so far identified in the aggregates of isomeric BChl c in solution and in the solid aggregate of an isomeric mixture of BChl c extracted from Chlorobium limicola. The selection was based on the comparison of the intermolecular (13)C...(13)C magnetic-dipole correlations with the nearest neighbor carbon-to-carbon close contacts simulated for the above six different stackings. It has turned out that the stacking of the macrocycles in chlorosome is basically the same as that in the in vitro solid aggregate. (2) The crucial role of water in stabilizing the aggregate structure in chlorosome was shown by tracing the dehydration processes and by comparison with the solid aggregate using the X-ray diffraction pattern. Possible binding sites of water molecules were located, by structural simulation, based on the particular stacking structure. (3) The dimer-based stacking of the macrocycles was evidenced by (25)Mg NMR spectroscopy, which exhibited a pair of signals showing different quadrupole coupling, due to the presence or absence of a water molecule in the axial position. PMID- 19123919 TI - Enzymatic mechanism of human apurinic/apyrimidinic endonuclease against a THF AP site model substrate. AB - The endonucleolytic activity of human apurinic/apyrimidinic endonuclease (AP endo) is a major factor in the maintenance of the integrity of the human genome. There are estimates that this enzyme is responsible for eliminating as many as 10(5) potentially mutagenic and genotoxic lesions from the genome of each cell every day. Furthermore, inhibition of AP endonuclease may be effective in decreasing the dose requirements of chemotherapeutics used in the treatment of cancer as well as other diseases. Therefore, it is essential to accurately and directly characterize the enzymatic mechanism of AP endo. Here we describe specifically designed double-stranded DNA oligomers containing tetrahydrofuran (THF) with a 5'-phosphorothioate linkage as the abasic site substrate. Using H(2)(18)O during the cleavage reaction and leveraging the stereochemical preferences of AP endo and T4 DNA ligase for phosphorothioate substrates, we show that AP endo acts by a one-step associative phosphoryl transfer mechanism on a THF-containing substrate. PMID- 19123921 TI - Contribution of individual amino acids to the RNA binding activity of the Wilms' tumor suppressor protein WT1. AB - In addition to binding to DNA, the zinc finger protein WT1 can also bind specifically to RNA. To determine the role of individual zinc fingers of the protein in this RNA binding activity, deletion and substitution mutants of the WT1 zinc finger domain were constructed. The effects of the various mutations on the binding of WT1 to the RNA aptamers RNA22 and RNA38 were determined using a quantitative equilibrium binding assay. The results indicate that zinc fingers 2 and 3 of WT1 are essential for the binding of the protein to the RNA aptamers. For both of these fingers, the arginine residue immediately preceding the alpha helix makes a significant contribution to RNA binding. For zinc finger 2, a second arginine residue within the alpha-helix is also critical for RNA binding, while several alpha-helical residues in zinc finger 3 contribute to the overall affinity of WT1 for RNA. Investigating the effects of the same point mutations on DNA binding indicates that there are similarities and differences in the contributions of zinc fingers 2 and 3 to the DNA and RNA binding activities of WT1. PMID- 19123923 TI - Tautomer enumeration and stability prediction for virtual screening on large chemical databases. AB - Tautomeric rearrangements affect the results of cheminformatics applications that depend on the knowledge of the 2D or 3D structure of a compound, such as tools for database searches, fingerprint generation, virtual screening, and physical chemical properties prediction. In this paper we present TauThor, a tool to enumerate tautomers and predict tautomer stability in the aqueous medium. The enumeration is based on a recursive process that generates tautomers according to the general scheme HX-Y=Z left harpoon over right harpoon X=Y-ZH. The stability of a tautomer is calculated by using a library of 145 fragments associated with experimental tautomeric percentages in water and a pK(a) based-method that utilizes pK(a) values predicted by MoKa. Predicted tautomeric ratios based on pK(a) calculations were benchmarked against literature data for a set of eleven compounds. The FDA approved drugs database, the NCI database and two vendor databases - Specs Screening Library and Asinex Gold Collection - were used to illustrate the impact of tautomerism on chemical libraries and to evaluate the relative occurrences of alternative tautomeric forms. PMID- 19123922 TI - C-Terminal membrane spanning region of human heme oxygenase-1 mediates a time dependent complex formation with cytochrome P450 reductase. AB - Heme oxygenase-1 (HO-1) catalyzes the oxidative degradation of heme to biliverdin, carbon monoxide, and free iron in a reaction requiring the interaction of HO-1 with NADPH-cytochrome P450 reductase (CPR). HO-1 is bound to the endoplasmic reticulum by 23 C-terminal amino acids; however, a soluble HO-1 (sHO-1) lacking this membrane spanning region has been extensively studied. The goal of this project was to characterize the effect of the C-terminal hydrophobic domain on formation of the HO-1/CPR complex. Full-length HO-1 was shown to exhibit higher reaction rates than sHO-1, particularly at subsaturating CPR, indicating that the C-terminal region influences HO-1 binding to CPR. The increased activity of HO-1 was attributable to a time-dependent formation of a low K(m) HO-1/CPR complex that was not seen with sHO1. Gel filtration analysis confirmed the formation of multiple high molecular weight complexes in the presence and absence of the synthetic lipid dilauroylphosphatidylcholine (DLPC). However, the largest complex appeared following a 2 h incubation of HO-1 and CPR in DLPC, suggesting that the C-terminal region was required for the high-affinity HO-1/CPR complex formation and membrane incorporation. These data demonstrate that the C-terminal region of HO-1 influenced complex formation and ultimately its affinity for CPR. PMID- 19123924 TI - How similar are similarity searching methods? A principal component analysis of molecular descriptor space. AB - Different molecular descriptors capture different aspects of molecular structures, but this effect has not yet been quantified systematically on a large scale. In this work, we calculate the similarity of 37 descriptors by repeatedly selecting query compounds and ranking the rest of the database. Euclidean distances between the rank-ordering of different descriptors are calculated to determine descriptor (as opposed to compound) similarity, followed by PCA for visualization. Four broad descriptor classes are identified, which are circular fingerprints; circular fingerprints considering counts; path-based and keyed fingerprints; and pharmacophoric descriptors. Descriptor behavior is much more defined by those four classes than the particular parametrization. Using counts instead of the presence/absence of fingerprints significantly changes descriptor behavior, which is crucial for performance of topological autocorrelation vectors, but not circular fingerprints. Four-point pharmacophores (piDAPH4) surprisingly lead to much higher retrieval rates than three-point pharmacophores (28.21% vs 19.15%) but still similar rank-ordering of compounds (retrieval of similar actives). Looking into individual rankings, circular fingerprints seem more appropriate than path-based fingerprints if complex ring systems or branching patterns are present; count-based fingerprints could be more suitable in databases with a large number of repeated subunits (amide bonds, sugar rings, terpenes). Information-based selection of diverse fingerprints for consensus scoring (ECFP4/TGD fingerprints) led only to marginal improvement over single fingerprint results. While it seems to be nontrivial to exploit orthogonal descriptor behavior to improve retrieval rates in consensus virtual screening, those descriptors still each retrieve different actives which corroborates the strategy of employing diverse descriptors individually in prospective virtual screening settings. PMID- 19123925 TI - CD146 expression is associated with a poor prognosis in human breast tumors and with enhanced motility in breast cancer cell lines. AB - INTRODUCTION: Metastasis is a complex process involving loss of adhesion, migration, invasion and proliferation of cancer cells. Cell adhesion molecules play a pivotal role in this phenomenon by regulating cell-cell and cell-matrix interactions. CD146 (MCAM) is associated with an advanced tumor stage in melanoma, prostate cancer and ovarian cancer. Studies of CD146 expression and function in breast cancer remain scarce except for a report concluding that CD146 could act as a tumor suppressor in breast carcinogenesis. METHODS: To resolve these apparent discrepancies in the role of CD146 in tumor cells, we looked at the association of CD146 expression with histoclinical features in human primary breast cancers using DNA and tissue microarrays. By flow cytometry, we characterized CD146 expression on different breast cancer cell lines. Using siRNA or shRNA technology, we studied functional consequences of CD146 downmodulation of MDA-MB-231 cells in migration assays. Wild-type, mock-transfected and downmodulated transfected cells were profiled using whole-genome DNA microarrays to identify genes whose expression was modified by CD146 downregulation. RESULTS: Microarray studies revealed the association of higher levels of CD146 with histoclinical features that belong to the basal cluster of human tumors. Expression of CD146 protein on epithelial cells was detected in a small subset of cancers with histoclinical features of basal tumors. CD146+ cell lines displayed a mesenchymal phenotype. Downmodulation of CD146 expression in the MDA-MB-231 cell line resulted in downmodulation of vimentin, as well as of a set of genes that include both genes associated with a poor prognosis in a variety of cancers and genes known to promote cell motility. In vitro functional assays revealed decreased migration abilities associated with decreased CD146 expression. CONCLUSIONS: In addition to its expression in the vascular compartment, CD146 is expressed on a subset of epithelial cells in malignant breast. CD146 may directly or indirectly contribute to tumor aggressiveness by promoting malignant cell motility. Changes in molecular signatures following downmodulation of CD146 expression suggest that CD146 downmodulation is associated with the reversal of several biological characteristics associated with epithelial to mesenchymal transition, and the phenomenon associated with the metastatic process. PMID- 19123926 TI - Pattern of drug utilization for treatment of uncomplicated malaria in urban Ghana following national treatment policy change to artemisinin-combination therapy. AB - BACKGROUND: Change of first-line treatment of uncomplicated malaria to artemisinin-combination therapy (ACT) is widespread in Africa. To expand knowledge of safety profiles of ACT, pharmacovigilance activities are included in the implementation process of therapy changes. Ghana implemented first-line therapy of artesunate-amodiaquine in 2005. Drug utilization data is an important component of determining drug safety, and this paper describes how anti-malarials were prescribed within a prospective pharmacovigilance study in Ghana following anti-malarial treatment policy change. METHODS: Patients with diagnosis of uncomplicated malaria were recruited from pharmacies of health facilities throughout Accra in a cohort-event monitoring study. The main drug utilization outcomes were the relation of patient age, gender, type of facility attended, mode of diagnosis and concomitant treatments to the anti-malarial regimen prescribed. Logistic regression was used to predict prescription of nationally recommended first-line therapy and concomitant prescription of antibiotics. RESULTS: The cohort comprised 2,831 patients. Curative regimens containing an artemisinin derivative were given to 90.8% (n = 2,574) of patients, although 33% (n = 936) of patients received an artemisinin-based monotherapy. Predictors of first-line therapy were laboratory-confirmed diagnosis, age >5 years, and attending a government facility. Analgesics and antibiotics were the most commonly prescribed concomitant medications, with a median of two co prescriptions per patient (range 1-9). Patients above 12 years were significantly less likely to have antibiotics co-prescribed than patients under five years; those prescribed non-artemisinin monotherapies were more likely to receive antibiotics. A dihydroartemisinin-amodiaquine combination was the most used therapy for children under five years of age (29.0%, n = 177). CONCLUSION: This study shows that though first-line therapy recommendations may change, clinical practice may still be affected by factors other than the decision or ability to diagnose malaria. Age, diagnostic confirmation and suspected concurrent conditions lead to benefit:risk assessments for individual patients by clinicians as to which anti-malarial treatment to prescribe. This has implications for adherence to policy changes aiming to implement effective use of ACT. These results should inform education of health professionals and rational drug use policies to reduce poly-pharmacy, and also suggest a potential positive impact of increased access to testing for malaria both within health facilities and in homes. PMID- 19123927 TI - Socioeconomic differences in lack of recreational walking among older adults: the role of neighbourhood and individual factors. AB - BACKGROUND: People with a low socioeconomic status (SES) are more likely to be physically inactive than their higher status counterparts, however, the mechanisms underlying this socioeconomic gradient in physical inactivity remain largely unknown. Our aims were (1) to investigate socioeconomic differences in recreational walking among older adults and (2) to examine to what extent neighbourhood perceptions and individual cognitions regarding regular physical activity can explain these differences. METHODS: Data were obtained by a large scale postal survey among a stratified sample of older adults (age 55-75 years) (N = 1994), residing in 147 neighbourhoods of Eindhoven and surrounding areas, in the Netherlands. Multilevel logistic regression analyses assessed associations between SES (i.e. education and income), perceptions of the social and physical neighbourhood environment, measures of individual cognitions derived from the Theory of Planned Behaviour (e.g. attitude, perceived behaviour control), and recreational walking for >/=10 minutes/week (no vs. yes). RESULTS: Participants in the lowest educational group (OR 1.67 (95% CI, 1.18-2.35)) and lowest income group (OR 1.40 (95% CI, 0.98-2.01)) were more likely to report no recreational walking than their higher status counterparts. The association between SES and recreational walking attenuated when neighbourhood aesthetics was included in the model, and largely reduced when individual cognitions were added to the model (with largest effects of attitude, and intention regarding regular physical activity). The assiation between poor neighbourhood aesthetics and no recreational walking attenuated to (borderline) insignificance when individual cognitions were taken into account. CONCLUSION: Both neighbourhood aesthetics and individual cognitions regarding physical activity contributed to the explanation of socioeconomic differences in no recreational walking. Neighbourhood aesthetics may explain the association between SES and recreational walking largely via individual cognitions towards physical activity. Intervention and policy strategies to reduce socioeconomic differences in lack of recreational walking among older adults would be most effective if they intervene on both neighbourhood perceptions as well as individual cognitions. PMID- 19123928 TI - Xenopus NM23-X4 regulates retinal gliogenesis through interaction with p27Xic1. AB - BACKGROUND: In Xenopus retinogenesis, p27Xic1, a Xenopus cyclin dependent kinase inhibitor, functions as a cell fate determinant in both gliogenesis and neurogenesis in a context dependent manner. This activity is essential for co ordination of determination and cell cycle regulation. However, very little is known about the mechanism regulating the context dependent choice between gliogenesis versus neurogenesis. RESULTS: We have identified NM23-X4, a NM23 family member, as a binding partner of p27Xic1. NM23-X4 is expressed at the periphery of the ciliary marginal zone of the Xenopus retina and the expression overlaps with p27Xic1 at the central side. Our in vivo functional analysis in Xenopus retina has shown that knockdown of NM23-X4 activates gliogenesis. Furthermore, co-overexpression of NM23-X4 with p27Xic1 results in the inhibition of p27Xic1-mediated gliogenesis, through direct interaction of NM23-X4 with the amino-terminal side of p27Xic1. This inhibitory effect on gliogenesis requires serine-150 and histidine-148, which correspond to the important residues for the kinase activities of NM23 family members. CONCLUSION: This study demonstrates that NM23-X4 functions as an inhibitor of p27Xic1-mediated gliogenesis in Xenopus retina and suggests that this activity contributes to the proper spatio-temporal regulation of gliogenesis. PMID- 19123930 TI - Design of FitFor2 study: the effects of an exercise program on insulin sensitivity and plasma glucose levels in pregnant women at high risk for gestational diabetes. AB - BACKGROUND: Pregnancy is a period in the life of women that is often associated with decreased daily physical activity and/or exercise. However, maintaining adequate levels of daily physical activity during pregnancy is important for mother and child. Studies suggest that moderate daily physical activity and exercise during pregnancy are associated with reductions in the risk of gestational diabetes mellitus (GDM). However, at present, physical activity is not routinely advised to pregnant women at risk for gestational diabetes in the Netherlands. In FitFor2-study we aim to assess whether an exercise program can improve insulin sensitivity and fasting plasma glucose levels of women at high risk for gestational diabetes, assuming that this will lower their risk of gestational diabetes. METHODS: The FitFor2-study is a randomised controlled trial. Women who visit one of the participating hospitals or midwifery practices and who are at risk for gestational diabetes are eligible to participate. After baseline measurement they are randomly allocated to in the intervention or control group. The intervention group receives an exercise program twice a week in addition to usual care. The exercise program consist of aerobic and strength exercises and takes place under close supervision of a physiotherapist. Data are collected at 15, 24 and 32 weeks of pregnancy and 12 weeks after delivery. Primary maternal outcome measures are fasting plasma glucose and relative increase in insulin resistance. Primary neonatal outcome is birth weight. Secondary outcome measures are: maternal serum triglycerides, HDL, cholesterol, HbA1c, maternal weight gain during pregnancy, maternal physical activity level, foetal growth. DISCUSSION: If the FitFor2 intervention program proves to be effective, obstetricians and midwives should refer women at risk for GDM to a special exercise program. Exercise programs for pregnant women under supervision of an experienced trainer are already available in the Netherlands, and these programs could be adjusted easily for this target group. Furthermore, the costs of these programs should be refunded by including them in the basic health care cost reimbursement schemes. TRIAL REGISTRATION: NTR1139. PMID- 19123929 TI - Heterogeneity in the developmental potential of motor neuron progenitors revealed by clonal analysis of single cells in vitro. AB - BACKGROUND: The differentiation of neural progenitors into distinct classes within the central nervous system occurs over an extended period during which cells become progressively restricted in their fates. In the developing spinal cord, Sonic Hedgehog (Shh) controls neural fates in a concentration-dependent manner by establishing discrete ventral progenitor domains characterized by specific combinations of transcription factors. It is unclear whether motor neuron progenitors can maintain their identities when expanded in vitro and whether their developmental potentials are restricted when exposed to defined extracellular signals. RESULTS: We have generated mice expressing the enhanced green fluorescent protein under the control of the Nkx6.1 promoter, enabling fluorescence-activated cell sorting (FACS), purification and culture of individual spinal progenitors at clonal density, and analysis of their progeny. We demonstrate that cells isolated after progenitor domains are established are heterogeneous with respect to maintaining their identity after in vitro expansion. Most Nkx6.1+ progenitors lose their ventral identity following several divisions in culture, whereas a small subset is able to maintain its identity. Thus, subtype-restricted progenitors from the Nkx6.1+ region are present in the ventral spinal cord, although at a lower frequency than expected. Clones that maintain a motor neuron identity assume a transcriptional profile characteristic of thoracic motor neurons, despite some having been isolated from non-thoracic regions initially. Exposure of progenitors to Bone Morphogenetic Protein-4 induces some dorsal cell type characteristics in their progeny, revealing that lineage-restricted progenitor subtypes are not fully committed to their fates. CONCLUSION: These findings support a model whereby continuous Shh signaling is required to maintain the identity of ventral progenitors isolated from the spinal cord, including motor neuron progenitors, after in vitro expansion. They also demonstrate that pre-patterned neural progenitors isolated from the central nervous system can change their regional identity in vitro to acquire a broader developmental potential. PMID- 19123931 TI - Physical characteristics of the back are not predictive of low back pain in healthy workers: a prospective study. AB - BACKGROUND: In the working population, back disorders are an important reason for sick leave and permanent work inability. In the context of fitting the job to the worker, one of the primary tasks of the occupational health physician is to evaluate the balance between work-related and individual variables. Since this evaluation of work capacity often consists of a physical examination of the back, the objective of this study was to investigate whether a physical examination of the low back, which is routinely performed in occupational medicine, predicts the development of low back pain (LBP). METHODS: This study is part of the Belgian Low Back Cohort (BelCoBack) Study, a prospective study to identify risk factors for the development of low back disorders in occupational settings. The study population for this paper were 692 young healthcare or distribution workers (mean age of 26 years) with no or limited back antecedents in the year before inclusion. At baseline, these workers underwent a standardised physical examination of the low back. One year later, they completed a questionnaire on the occurrence of LBP and some of its characteristics. To study the respective role of predictors at baseline on the occurrence of LBP, we opted for Cox regression with a constant risk period. Analyses were performed separately for workers without any back antecedents in the year before inclusion ('asymptomatic' workers) and for workers with limited back antecedents in the year before inclusion ('mildly symptomatic' workers). RESULTS: In the group of 'asymptomatic' workers, obese workers showed a more than twofold-increased risk on the development of LBP as compared to non-obese colleagues (RR 2.57, 95%CI: 1.09 - 6.09). In the group of 'mildly symptomatic' workers, the self-reports of pain before the examination turned out to be most predictive (RR 3.89, 95%CI: 1.20 - 12.64). CONCLUSION: This study showed that, in a population of young workers wh no or limited antecedents of LBP at baseline, physical examinations, as routinely assessed in occupational medicine, are not useful to predict workers at risk for the development of back disorders one year later. PMID- 19123932 TI - Rapid decrease of malaria morbidity following the introduction of community-based monitoring in a rural area of central Vietnam. AB - BACKGROUND: Despite a successful control programme, malaria has not completely disappeared in Vietnam; it remains endemic in remote areas of central Vietnam, where standard control activities seem to be less effective. The evolution of malaria prevalence and incidence over two and half years in a rural area of central Vietnam, after the introduction of community-based monitoring of malaria cases, is presented. METHODS: After a complete census, six cross-sectional surveys and passive detection of malaria cases (by village and commune health workers using rapid diagnostic tests) were carried out between March 2004 and December 2006 in Ninh-Thuan province, in a population of about 10,000 individuals. The prevalence of malaria infection and the incidence of clinical cases were estimated. RESULTS: Malaria prevalence significantly decreased from 13.6% (281/2,068) in December 2004 to 4.0% (80/2,019) in December 2006. Plasmodium falciparum and Plasmodium vivax were the most common infections with few Plasmodium malariae mono-infections and some mixed infections. During the study period, malaria incidence decreased by more than 50%, from 25.7/1,000 population at risk in the second half of 2004 to 12.3/1,000 in the second half of 2006. The incidence showed seasonal variations, with a yearly peak between June and December, except in 2006 when the peak observed in the previous years did not occur. CONCLUSION: Over a 2.5-year follow-up period, malaria prevalence and incidence decreased by more than 70% and 50%, respectively. Possibly, this could be attributed to the setting up of a passive case detection system based on village health workers, indicating that a major impact on the malaria burden can be obtained whenever prompt diagnosis and adequate treatment are available. PMID- 19123934 TI - Low-intensity body building exercise induced rhabdomyolysis: a case report. AB - INTRODUCTION: Rhabdomyolysis is a severe and debilitating condition that promotes muscle breakdown and is a relatively rare, not always diagnosed cause of acute renal failure (ARF) with an 8-20% reported incidence. Exertional rhabdomyolysis only appears in adult patients 24-48 h after strenuous activities as military basic training, weight lifting, and marathon running. CASE PRESENTATION: A 30 year-old man was admitted to our department because of weakness and painful swelling of the muscles as well as dark urine appearing 24 h after carrying out a body-building exercises of low intensity. The development of an acute exertional rhabdomyolysis was confirmed by the increased serum enzyme levels and myoglobinuria. The patient was treated with intravenous sodium chloride, and sodium bicarbonate. The nephrotoxicity of myoglobin was decreased by forced alkaline diuresis. CONCLUSION: The reported case emphasizes the occurrence of acute rhabdomyolysis even in those who underwent a low-intensity exercise. A proper treatment is mandatory to avoid a sudden worsening of clinical conditions eventually evolving to acute renal failure. PMID- 19123933 TI - Spontaneous improvement in randomised clinical trials: meta-analysis of three armed trials comparing no treatment, placebo and active intervention. AB - BACKGROUND: It can be challenging for patients and clinicians to properly interpret a change in the clinical condition after a treatment has been given. It is not known to which extent spontaneous improvement, effect of placebo and effect of active interventions contribute to the observed change from baseline, and we aimed at quantifying these contributions. METHODS: Systematic review and meta-analysis, based on a Cochrane review of the effect of placebo interventions for all clinical conditions. We selected all trials that had randomised the patients to three arms: no treatment, placebo and active intervention, and that had used an outcome that was measured on a continuous scale or on a ranking scale. Clinical conditions that had been studied in less than three trials were excluded. RESULTS: We analysed 37 trials (2900 patients) that covered 8 clinical conditions. The active interventions were psychological in 17 trials, physical in 15 trials, and pharmacological in 5 trials. Overall, across all conditions and interventions, there was a statistically significant change from baseline in all three arms. The standardized mean difference (SMD) for change from baseline was 0.24 (95% confidence interval -0.36 to -0.12) for no treatment, -0.44 (-0.61 to 0.28) for placebo, and -1.01 (-1.16 to -0.86) for active treatment. Thus, on average, the relative contributions of spontaneous improvement and of placebo to that of the active interventions were 24% and 20%, respectively, but with some uncertainty, as indicated by the confidence intervals for the three SMDs. The conditions that had the most pronounced spontaneous improvement were nausea (45%), smoking (40%), depression (35%), phobia (34%) and acute pain (25%). CONCLUSION: Spontaneous improvement and effect of placebo contributed importantly to the observed treatment effect in actively treated patients, but the relative importance of these factors differed according to clinical condition and intervention. PMID- 19123935 TI - Hepatic splenosis mimicking HCC in a patient with hepatitis C liver cirrhosis and mildly raised alpha feto protein; the important role of explorative laparoscopy. AB - BACKGROUND: Splenosis is a heterotropic implantation of splenic fragments onto exposed vascularised peritoneal and intrathoracic surfaces, following splenic injury or elective splenectomy. CASE PRESENTATION: A 60 year old cirrhotic patient was referred to us with a hepatic mass, suspected to be HCC in a cirrhotic liver. A computerized tomography scan (CT) demonstrated a cirrhotic liver with a 2 x 2.7 cm focal hypervascular nodule, lying peripherally at the junction of segment 7 and 8. Diagnostic laparoscopy demonstrated a 3 cm exofitic dark brown splenunculus attached to the diaphragm and indenting the surface of segment 7 of the liver. The lesion was easily resected laparoscopically and shaved from the live surface with no need for a liver resection. The histopathological assessment confirmed the diagnosis of splenunculus, with no evidence of neoplasia. CONCLUSION: Hepatic splenosis is not a rare event and should be suspected in patients with a history of splenic trauma or splenectomy. Correct diagnosis is essential and will determine subsequent management plans. In doubtful cases laparoscopic investigation can offere essential information and should be part of the standard protocol for investigating suspected splenosis. PMID- 19123936 TI - Gastric outlet obstruction possibly secondary to ulceration in a 2-year-old girl: a case report. AB - Gastric outlet obstruction due to ulceration is extremely rare in childhood. We report a case of gastric outlet obstruction possibly secondary to peptic ulceration and our surgical management. Our approach, without vagotomy or antrectomy, would appear to be a safe and effective. PMID- 19123938 TI - Is a purpose of REM sleep atonia to help regenerate intervertebral disc volumetric loss? AB - The nature of atonia in sleep continues to be enigmatic. This article discusses a new hypothesis for complete core muscle relaxation in REM sleep, suggesting a bottom-up recuperative perspective. That is, does the atonia in REM sleep provide a utility to help restore the mechanobiology and respective diurnal intervertebral disc hydraulic loss? By combining the effects of gravity with current compressive concepts in spinal stability, this article looks at vertebral approximation as a deleterious experience with an intrinsic biological need to keep vertebrae separated. Methods using polysomnography and recumbent MRI are discussed. PMID- 19123937 TI - Comparative analysis of processed ribosomal protein pseudogenes in four mammalian genomes. AB - BACKGROUND: The availability of genome sequences of numerous organisms allows comparative study of pseudogenes in syntenic regions. Conservation of pseudogenes suggests that they might have a functional role in some instances. RESULTS: We report the first large-scale comparative analysis of ribosomal protein pseudogenes in four mammalian genomes (human, chimpanzee, mouse and rat). To this end, we have assigned these pseudogenes in the four organisms using an automated pipeline and make the results available online. Each organism has a large number of ribosomal protein pseudogenes (approximately 1,400 to 2,800). The majority of them are processed (generated by retrotransposition). However, we do not see a correlation between the number of pseudogenes associated with a ribosomal protein gene and its mRNA abundance. Analysis of pseudogenes in syntenic regions between species shows that most are conserved between human and chimpanzee, but very few are conserved between primates and rodents. Interestingly, syntenic pseudogenes have a lower rate of nucleotide substitution than their surrounding intergenic DNA. Moreover, evidence from expressed sequence tags indicates that two pseudogenes conserved between human and mouse are transcribed. Detailed analysis shows that one of them, the pseudogene of RPS27, is likely to be a protein-coding gene. This is significant as previous reports indicated there are exactly 80 ribosomal protein genes encoded by the human genome. CONCLUSIONS: Our analysis indicates that processed ribosomal protein pseudogenes abound in mammalian genomes, but few of these are conserved between primates and rodents. This highlights the large amount of recent retrotranspositional activity in mammals and a relatively larger amount of it in the rodent lineage. PMID- 19123939 TI - Patterns of seasonal phytoplankton distribution in prairie saline lakes of the northern Great Plains (U.S.A.). AB - Seasonal changes in freshwater phytoplankton communities have been extensively studied, but key drivers of phytoplankton in saline lakes are currently not well understood. Comparative lake studies of 19 prairie saline lakes in the northern Great Plains (USA) were conducted in spring and summer of 2004, with data gathered for a suite of limnological parameters. Nutrient enrichment assays for natural phytoplankton assemblages were also performed in spring and summer of 2006. Canonical correspondence analysis of 2004 data showed salinity (logCl), nitrogen, and phosphorus (N:P ratios) to be the main drivers of phytoplankton distribution in the spring, and phosphorus (C:P ratios), iron (logTFe), and nitrogen (logTN) as important factors in the summer. Despite major differences in nutrient limitation patterns (P-limitation in freshwater systems, N-limitation in saline systems), seasonal patterns of phytoplankton phyla changes in these saline lakes were similar to those of freshwater systems. Dominance shifted from diatoms in the spring to cyanobacteria in the summer. Nutrient enrichment assays (control, +Fe, +N, +P, +N+P) in 2006 indicated that nutrient limitation is generally more consistent within lakes than for individual taxa across systems, with widespread nitrogen and secondary phosphorus limitation. Understanding phytoplankton community structure provides insight into the overall ecology of saline lakes, and will assist in the future conservation and management of these valuable and climatically-sensitive systems. PMID- 19123940 TI - Evolution and differential expression of a vertebrate vitellogenin gene cluster. AB - BACKGROUND: The multiplicity or loss of the vitellogenin (vtg) gene family in vertebrates has been argued to have broad implications for the mode of reproduction (placental or non-placental), cleavage pattern (meroblastic or holoblastic) and character of the egg (pelagic or benthic). Earlier proposals for the existence of three forms of vertebrate vtgs present conflicting models for their origin and subsequent duplication. RESULTS: By integrating phylogenetics of novel vtg transcripts from old and modern teleosts with syntenic analyses of all available genomic variants of non-metatherian vertebrates we identify the gene orthologies between the Sarcopterygii (tetrapod branch) and Actinopterygii (fish branch). We argue that the vertebrate vtg gene cluster originated in proto chromosome m, but that vtg genes have subsequently duplicated and rearranged following whole genome duplications. Sequencing of a novel fourth vtg transcript in labrid species, and the presence of duplicated paralogs in certain model organisms supports the notion that lineage-specific gene duplications frequently occur in teleosts. The data show that the vtg gene cluster is more conserved between acanthomorph teleosts and tetrapods, than in ostariophysan teleosts such as the zebrafish. The differential expression of the labrid vtg genes are further consistent with the notion that neofunctionalized Aa-type vtgs are important determinants of the pelagic or benthic character of the eggs in acanthomorph teleosts. CONCLUSION: The vertebrate vtg gene cluster existed prior to the separation of Sarcopterygii from Actinopterygii >450 million years ago, a period associated with the second round of whole genome duplication. The presence of higher copy numbers in a more highly expressed subcluster is particularly prevalent in teleosts. The differential expression and latent neofunctionalization of vtg genes in acanthomorph teleosts is an adaptive feature associated with oocyte hydration and spawning in the marine environment. PMID- 19123942 TI - Health promotion and the randomised controlled trial: a square peg in a round hole? AB - In their paper published in BMC Oral Health in March, Barker and Horton present qualitative data which explored Latino parents' main concerns regarding accessing dental care for their pre-school children. In the radical discourse of health promotion the use of participant narratives is a first and essential step in community development interventions. While there is agreement regarding the development and implementation of health promotion, the means by which it is evaluated or the type of evaluation design used, is hotly debated. This commentary outlines the rationale of adopting a randomised controlled trial methodology, contrasts it with realistic evaluation and considers design evaluation in the light of the Medical Research Council's (MRC) guidance of 2000 and 2008. It is at this juncture that the commentary suggests that, despite the MRC's acknowledgement of the limitations of its 2000 guidance, there remains, in the 2008 guidance, an underlying insistence to use design evaluations which control for selection bias and confounding extraneous factors. For the evaluation of health promotion interventions it may remain a case of fitting a square peg into a round hole. PMID- 19123941 TI - Transcript and proteomic analysis of developing white lupin (Lupinus albus L.) roots. AB - BACKGROUND: White lupin (Lupinus albus L.) roots efficiently take up and accumulate (heavy) metals, adapt to phosphate deficiency by forming cluster roots, and secrete antimicrobial prenylated isoflavones during development. Genomic and proteomic approaches were applied to identify candidate genes and proteins involved in antimicrobial defense and (heavy) metal uptake and translocation. RESULTS: A cDNA library was constructed from roots of white lupin seedlings. Eight thousand clones were randomly sequenced and assembled into 2,455 unigenes, which were annotated based on homologous matches in the NCBInr protein database. A reference map of developing white lupin root proteins was established through 2-D gel electrophoresis and peptide mass fingerprinting. High quality peptide mass spectra were obtained for 170 proteins. Microsomal membrane proteins were separated by 1-D gel electrophoresis and identified by LC-MS/MS. A total of 74 proteins were putatively identified by the peptide mass fingerprinting and the LC-MS/MS methods. Genomic and proteomic analyses identified candidate genes and proteins encoding metal binding and/or transport proteins, transcription factors, ABC transporters and phenylpropanoid biosynthetic enzymes. CONCLUSION: The combined EST and protein datasets will facilitate the understanding of white lupin's response to biotic and abiotic stresses and its utility for phytoremediation. The root ESTs provided 82 perfect simple sequence repeat (SSR) markers with potential utility in breeding white lupin for enhanced agronomic traits. PMID- 19123944 TI - Cytokinesis is blocked in mammalian cells transfected with Chlamydia trachomatis gene CT223. AB - BACKGROUND: The chlamydiae alter many aspects of host cell biology, including the division process, but the molecular biology of these alterations remains poorly characterized. Chlamydial inclusion membrane proteins (Incs) are likely candidates for direct interactions with host cell cytosolic proteins, as they are secreted to the inclusion membrane and exposed to the cytosol. The inc gene CT223 is one of a sequential set of orfs that encode or are predicted to encode Inc proteins. CT223p is localized to the inclusion membrane in all tested C. trachomatis serovars. RESULTS: A plasmid transfection approach was used to examine the function of the product of CT223 and other Inc proteins within uninfected mammalian cells. Fluorescence microscopy was used to demonstrate that CT223, and, to a lesser extent, adjacent inc genes, are capable of blocking host cell cytokinesis and facilitating centromere supranumeracy defects seen by others in chlamydiae-infected cells. Both phenotypes were associated with transfection of plasmids encoding the carboxy-terminal tail of CT223p, a region of the protein that is likely exposed to the cytosol in infected cells. CONCLUSION: These studies suggest that certain Inc proteins block cytokinesis in C. trachomatis infected cells. These results are consistent with the work of others showing chlamydial inhibition of host cell cytokinesis. PMID- 19123945 TI - Mapping new theoretical and methodological terrain for knowledge translation: contributions from critical realism and the arts. AB - BACKGROUND: Clinical practice guidelines have been a popular tool for the improvement of health care through the implementation of evidence from systematic research. Yet, it is increasingly clear that knowledge alone is insufficient to change practice. The social, cultural, and material contexts within which practice occurs may invite or reject innovation, complement or inhibit the activities required for success, and sustain or alter adherence to entrenched practices. However, knowledge translation (KT) models are limited in providing insight about how and why contextual contingencies interact, the causal mechanisms linking structural aspects of context and individual agency, and how these mechanisms influence KT. Another limitation of KT models is the neglect of methods to engage potential adopters of the innovation in critical reflection about aspects of context that influence practice, the relevance and meaning of innovation in the context of practice, and the identification of strategies for bringing about meaningful change. DISCUSSION: This paper presents a KT model, the Critical Realism and the Arts Research Utilization Model (CRARUM), that combines critical realism and arts-based methodologies. Critical realism facilitates understanding of clinical settings by providing insight into the interrelationship between its structures and potentials, and individual action. The arts nurture empathy, and can foster reflection on the ways in which contextual factors influence and shape clinical practice, and how they may facilitate or impede change. The combination of critical realism and the arts within the CRARUM model promotes the successful embedding of interventions, and greater impact and sustainability. CONCLUSION: CRARUM has the potential to strengthen the science of implementation research by addressing the complexities of practice settings, and engaging potential adopters to critically reflect on existing and proposed practices and strategies for sustaining change. PMID- 19123943 TI - Activation of the steroid and xenobiotic receptor, SXR, induces apoptosis in breast cancer cells. AB - BACKGROUND: The steroid and xenobiotic receptor, SXR, is an orphan nuclear receptor that regulates metabolism of diverse dietary, endobiotic, and xenobiotic compounds. SXR is expressed at high levels in the liver and intestine, and at lower levels in breast and other tissues where its function was unknown. Since many breast cancer preventive and therapeutic compounds are SXR activators, we hypothesized that some beneficial effects of these compounds are mediated through SXR. METHODS: To test this hypothesis, we measured proliferation of breast cancer cells in response to SXR activators and evaluated consequent changes in the expression of genes critical for proliferation and cell-cycle control using quantitative RT-PCR and western blotting. Results were confirmed using siRNA mediated gene knockdown. Statistical analysis was by t-test or ANOVA and a P value < or = 0.05 was considered to be significant. RESULTS: Many structurally and functionally distinct SXR activators inhibited the proliferation of MCF-7 and ZR-75-1 breast cancer cells by inducing cell cycle arrest at the G1/S phase followed by apoptosis. Decreased growth in response to SXR activation was associated with stabilization of p53 and up-regulation of cell cycle regulatory and pro-apoptotic genes such as p21, PUMA and BAX. These gene expression changes were preceded by an increase in inducible nitric oxide synthase and nitric oxide in these cells. Inhibition of iNOS blocked the induction of p53. p53 knockdown inhibited up-regulation of p21 and BAX. We infer that NO is required for p53 induction and that p53 is required for up-regulation of cell cycle regulatory and apoptotic genes in this system. SXR activator-induced increases in iNOS levels were inhibited by siRNA-mediated knockdown of SXR, indicating that SXR activation is necessary for subsequent regulation of iNOS expression. CONCLUSION: We conclude that activation of SXR is anti-proliferative in p53 wild type breast cancer cells and that this effect is mechanistically dependent upon the local production of NO and NO-dependent up-regulation of p53. These findings reveal a novel biological function for SXR and suggest that a subset of SXR activators may function as effective therapeutic and chemo-preventative agents for certain types of breast cancers. PMID- 19123946 TI - Gene expression profiling of whole blood: comparison of target preparation methods for accurate and reproducible microarray analysis. AB - BACKGROUND: Peripheral blood is an accessible and informative source of transcriptomal information for many human disease and pharmacogenomic studies. While there can be significant advantages to analyzing RNA isolated from whole blood, particularly in clinical studies, the preparation of samples for microarray analysis is complicated by the need to minimize artifacts associated with highly abundant globin RNA transcripts. The impact of globin RNA transcripts on expression profiling data can potentially be reduced by using RNA preparation and labeling methods that remove or block globin RNA during the microarray assay. We compared four different methods for preparing microarray hybridization targets from human whole blood collected in PAXGene tubes. Three of the methods utilized the Affymetrix one-cycle cDNA synthesis/in vitro transcription protocol but varied treatment of input RNA as follows: i. no treatment; ii. treatment with GLOBINclear; or iii. treatment with globin PNA oligos. In the fourth method cDNA targets were prepared with the Ovation amplification and labeling system. RESULTS: We find that microarray targets generated with labeling methods that reduce globin mRNA levels or minimize the impact of globin transcripts during hybridization detect more transcripts in the microarray assay compared with the standard Affymetrix method. Comparison of microarray results with quantitative PCR analysis of a panel of genes from the NF-kappa B pathway shows good correlation of transcript measurements produced with all four target preparation methods, although method-specific differences in overall correlation were observed. The impact of freezing blood collected in PAXGene tubes on data reproducibility was also examined. Expression profiles show little or no difference when RNA is extracted from either fresh or frozen blood samples. CONCLUSION: RNA preparation and labeling methods designed to reduce the impact of globin mRNA transcripts can significantly improve the sensitivity of the DNA microarray expression profiling assay for whole blood samples. While blockage of globin transcripts during first strand cDNA synthesis with globin PNAs resulted in the best overall performance in this study, we conclude that selection of a protocol for expression profiling studies in blood should depend on several factors, including implementation requirements of the method and study design. RNA isolated from either freshly collected or frozen blood samples stored in PAXGene tubes can be used without altering gene expression profiles. PMID- 19123948 TI - Acute central retinal artery occlusion presenting as CREST syndrome: a case report. AB - BACKGROUND: A 75 year old lady presented with acute central retinal artery occlusion and contralateral cotton wool spots. CASE PRESENTATION: General physical examination and investigations led to a diagnosis of CREST syndrome; however, association of central retinal artery occlusion with CREST syndrome is not well known. While diabetes, systemic hypertension, carotid atherosclerosis and cardiac pathology are common causes of CRAO it is always important to rule out giant cell arteritis. CONCLUSION: This case highlights that inflammatory causes of central retinal artery occlusion other than giant cell arteritis should also be considered as a possibility to spare unnecessary use of excessive systemic corticosteroids. PMID- 19123947 TI - Genome sequence analysis of Helicobacter pylori strains associated with gastric ulceration and gastric cancer. AB - BACKGROUND: Persistent colonization of the human stomach by Helicobacter pylori is associated with asymptomatic gastric inflammation (gastritis) and an increased risk of duodenal ulceration, gastric ulceration, and non-cardia gastric cancer. In previous studies, the genome sequences of H. pylori strains from patients with gastritis or duodenal ulcer disease have been analyzed. In this study, we analyzed the genome sequences of an H. pylori strain (98-10) isolated from a patient with gastric cancer and an H. pylori strain (B128) isolated from a patient with gastric ulcer disease. RESULTS: Based on multilocus sequence typing, strain 98-10 was most closely related to H. pylori strains of East Asian origin and strain B128 was most closely related to strains of European origin. Strain 98 10 contained multiple features characteristic of East Asian strains, including a type s1c vacA allele and a cagA allele encoding an EPIYA-D tyrosine phosphorylation motif. A core genome of 1237 genes was present in all five strains for which genome sequences were available. Among the 1237 core genes, a subset of alleles was highly divergent in the East Asian strain 98-10, encoding proteins that exhibited <90% amino acid sequence identity compared to corresponding proteins in the other four strains. Unique strain-specific genes were identified in each of the newly sequenced strains, and a set of strain specific genes was shared among H. pylori strains associated with gastric cancer or premalignant gastric lesions. CONCLUSION: These data provide insight into the diversity that exists among H. pylori strains from diverse clinical and geographic origins. Highly divergent alleles and strain-specific genes identified in this study may represent useful biomarkers for analyzing geographic partitioning of H. pylori and for identifying strains capable of inducing malignant or premalignant gastric lesions. PMID- 19123949 TI - Elective laparoscopic splenectomy for giant hemangioma: a case report. AB - Although unusual, hemangioma is the most common primary splenic neoplasm. Splenectomy is indicated when the tumor is large, with increased risk of hemorrhage. The laparoscopic approach is preferred for most elective splenectomies. Although technically feasible, laparoscopic splenectomy can be a challenge in the patient with splenomegaly. We present herein a case of an 18 year-old male asymptomatic patient who underwent laparoscopic splenectomy for the incidental finding of splenomegaly caused by a large splenic hemangioma. Laparoscopic splenectomy appears to be a safe and effective procedure, in appropriately experienced hands, for patients with splenomegaly, given the spleen's fragile anatomy and its relationship to other abdominal viscera. PMID- 19123950 TI - HER-2/neu amplification testing in breast cancer by Multiplex Ligation-dependent Probe Amplification: influence of manual- and laser microdissection. AB - BACKGROUND: Accurate assessment of HER-2/neu status is crucial for proper prognostic information and to offer direct appropriate treatment for breast cancer patients. Next to immunohistochemistry (IHC) to evaluate HER2 protein overexpression, a second line gene amplification test is generally deemed necessary for cases with equivocal protein expression. Recently, a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), was introduced as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer. MLPA was previously shown to correlate well with IHC and in situ hybridization (ISH), but a low tumor percentage in the tissue tested could negatively affect the accuracy of MLPA results. METHODS: To examine this, MLPA was repeated in 42 patients after serial H&E section guided manual dissection with a scalpel and after laser microdissection of the tumor. RESULTS: Both dissection techniques led to higher HER2 gene copy number ratios and thereby made MLPA more quantitative. Concordance between MLPA and ISH improved from 61% to 84% after manual microdissection and to 90% after laser microdissection. CONCLUSION: Manual and laser microdissection similarly increase the dynamic range of MLPA copy number ratios which is a technical advantage. As clinically a dichotomization between normal and amplified suffices and MLPA is relatively unsensitive to tumor content, microdissection before MLPA may not be routinely necessary but may be advisable in case of very low tumor content ( 75% overall radiochemical yield and > 99% radiochemical purity. [125I]IBDG was evaluated as a substrate using beta-galactosidase-expressing (D54L) and nonexpressing (D54) human glioma cell lines. A 24-hour incubation of this substrate with cultured cells revealed a 6.5-fold greater intracellular trapping of radioactivity in D54L cells compared with D54 cells. Systemic delivery of [125I]IBDG in nude mice bearing D54L tumors failed to show significant trapping of radioactivity within these tumors by SPECT imaging. In contrast, intratumoral injection of the substrate resulted in efficient trapping of radioactivity in D54L tumors but not D54 tumors, resulting in clear SPECT visualization of the former tumor. Based on dynamic SPECT imaging and blood metabolite analysis, we conclude that although [125I]IBDG is an efficient in vivo substrate for beta galactosidase, its rapid renal clearance hampers its intratumoral availability on systemic administration. PMID- 19123991 TI - Optimizing imaging of three-dimensional multicellular tumor spheroids with fluorescent reporter proteins using confocal microscopy. AB - Tumor spheroids more faithfully mimic tumor biology than monolayer cultures and require three-dimensional microscopy. Our goal in this study was to overcome the limitations of signal to noise ratio that have traditionally limited three dimensional imaging to depths of 100 microm or less. We studied the expression of hypoxia-inducible factor 1alpha (HIF-1alpha), the main regulator of cellular hypoxic response in C6 glioma spheroids. In our spheroids, red fluorescent protein is expressed constitutively and green fluorescent protein is expressed conditionally under control of a HIF-1alpha promoter. In this article, we show a series of optimizations that allowed us to obtain excellent quality confocal microscopy images at imaging depths of up to 320 microm. The combined use of special objectives, glass-bottomed culture dishes, and depth-dependent laser output modulation extended our depth range beyond previously accepted limits. This allowed us to image up to the equator of spheroids of 650 microm diameter, allowing interrogation of HIF-1alpha expression from the spheroid periphery to its hypoxic center. PMID- 19123990 TI - Functional live-cell imaging demonstrates that beta1-integrin promotes type IV collagen degradation by breast and prostate cancer cells. AB - The ability of tumor cells to adhere to, migrate on, and remodel extracellular matrices is mediated by cell surface receptors such as beta1-integrins. Here we conducted functional live-cell imaging in real time to investigate the effects of modulating beta1-integrin expression and function on proteolytic remodeling of the extracellular matrix. Human breast and prostate cancer cells were grown on reconstituted basement membrane containing a quenched fluorescent form of collagen IV. Generation of cleavage products and the resulting increases in fluorescence were imaged and quantified. Decreases in the expression and activity of beta1-integrin reduced digestion of quenched fluorescent-collagen IV by the breast and prostate cancer cells and correspondingly their invasion through and migration on reconstituted basement membrane. Decreased extracellular matrix degradation also was associated with changes in the constituents of proteolytic pathways: decreases in secretion of the cysteine protease cathepsin B, the matrix metalloproteinase (MMP)-13, and tissue inhibitors of metalloproteinases (TIMP)-1 and 2; a decrease in expression of MMP-14 or membrane type 1 MMP; and an increase in secretion of TIMP-3. This is the first study to demonstrate through functional live-cell imaging that downregulation of beta1-integrin expression and function reduces proteolysis of collagen IV by breast and prostate cancer cells. PMID- 19123992 TI - Stably integrated luxCDABE for assessment of Salmonella invasion kinetics. AB - Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence-based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent. PMID- 19123993 TI - Dynamic observation of the three-dimensional distribution of labeled liposomes using the novel high-resolution single-photon emission computed tomography. AB - The aim of this study was to show that the multipinhole technique (high resolution single-photon emission computed tomography [HiSPECT]) is suitable for dynamic imaging of both biodistribution and temporal uptake behavior of radiolabeled cationic liposomes in Balb/c-mice. HiSPECT uses multipinhole collimators adapted to a clinical SPECT scanner, together with a dedicated iterative reconstruction program. This technique provides both high spatial resolution and an improvement in sensitivity. Six male Balb/c mice received 9.8 +/- 4.0 MBq of the In 111-labeled liposomes. The measurements started directly after the injection and tomographic data were acquired in steps of 5 minutes. The regional evaluation displayed a high initial uptake of liposomes in the lungs (45.4%), which decreased to 25.1% after 30 minutes and to below 2% after 48 hours. In contrast, liver uptake increased in the first 30 minutes from 13.1 to 21.2% and remained relatively stable at 24.4% (24 hours) and 18.8% (48 hours). The data are interpreted as a slow shift of liposomes from the lungs into the liver and later to other organs such as the spleen and bladder. This study shows that the HiSPECT technique is capable of dynamically visualizing the uptake behavior of radioactively labeled liposomes in vivo with high temporal and spatial resolution. PMID- 19123994 TI - In vivo tumor targeting by the B-subunit of shiga toxin. AB - Delivery of drugs to the appropriate target cells would improve efficacy and reduce potential side effects. The nontoxic B-subunit of the intestinal pathogen produced Shiga toxin (STxB) binds specifically to the glycosphingolipid Gb3, overexpressed in membranes of certain tumor cells, and enters these cells through the retrograde pathway. Therefore, STxB binding to Gb3 receptors may be useful for cell-specific vectorization or imaging purposes. Here we labeled STxB with a fluorophore to evaluate its potential as an in vivo cell-specific targeting reagent in two different models of human colorectal carcinoma. Fluorescent STxB was administered systemically to xenografted nude mice, and its biodistribution was studied by optical imaging. The use of fluorescent STxB allowed the combination of the macroscopic observations with analyses at the cellular level using confocal microscopy. After administration, the fluorescent STxB was slowly eliminated by renal excretion. However, it accumulated in the tumor area. Furthermore, STxB was demonstrated to enter the Gb3-expressing tumoral cells, as well as the epithelial cells of the neovascularization and the monocytes and macrophages surrounding the xenografts. PMID- 19123995 TI - Dynamic analysis of the blood-brain barrier disruption in experimental stroke using time domain in vivo fluorescence imaging. AB - The blood-brain barrier (BBB) disruption following cerebral ischemia can be exploited to deliver imaging agents and therapeutics into the brain. The aim of this study was (a) to establish novel in vivo optical imaging methods for longitudinal assessment of the BBB disruption and (b) to assess size selectivity and temporal patterns of the BBB disruption after a transient focal ischemia. The BBB permeability was assessed using in vivo time domain near-infrared optical imaging after contrast enhancement with either free Cy5.5 (1 kDa) or Cy5.5 conjugated with bovine serum albumin (BSA) (67 kDa) in mice subjected to either 60- or 20-minute transient middle cerebral artery occlusion (MCAO) and various times of reperfusion (up to 14 days). In vivo imaging observations were corroborated by ex vivo brain imaging and microscopic analyses of fluorescent tracer extravasation. The in vivo optical contrast enhancement with Cy5.5 was spatially larger than that observed with BSA-Cy5.5. Longitudinal studies after a transient 20-minute MCAO suggested a bilateral BBB disruption, more pronounced in the ipsilateral hemisphere, peaking at day 7 and resolving at day 14 after ischemia. The area differential between the BBB disruption for small and large molecules could potentially be useful as a surrogate imaging marker for assessing perinfarct tissues to which neuroprotective therapies of appropriate sizes could be delivered. PMID- 19123996 TI - First In Vivo SPECT Imaging of Mouse Femorotibial Cartilage Using 99mTc-NTP 15-5. AB - This study aimed to report the first single-photon emission computed tomographic (SPECT) imaging of articular cartilage in mice using 99mTc-NTP 15-5 radiotracer. Mice intravenously injected with 99mTc-NTP 15-5 were submitted to (1) dynamic planar imaging, (2) static planar imaging, (3) 1 mm pinhole SPECT acquisition, and (4) dissection. Tomographic reconstruction of SPECT data was performed with a three-dimensional ordered subset expectation maximization algorithm, and slices were reconstructed in three axes. 99mTc-NTP 15-5 rapidly accumulated in the joint, with a peak of radioactivity being reached from 5 minutes postinjection and maintained for at least 90 minutes. Given that bone and muscle did not show any accumulation of the tracer, highly contrasted joint imaging was obtained from 15 minutes postinjection. When 1 mm pinhole SPECT acquisition was focused on the knee, the medial and lateral compartments of both the femoral condyle and tibial plateau were highly delineated, allowing a separate quantitation of tracer accumulation within each component of the femorotibial joint. A good correlation was found between tracer uptake determined by region of interest analysis of both planar and SPECT scans and dissection. This new approach to imaging of cartilage in mice provides joint functionality assessment in vivo, giving a unique opportunity to achieve a greater understanding of cartilage physiology in health and disease. PMID- 19123997 TI - In vivo biodistribution and lifetime analysis of cy5.5-conjugated rituximab in mice bearing lymphoid tumor xenograft using time-domain near-infrared optical imaging. AB - Rituximab is a chimeric monoclonal antibody directed against human CD20 antigen, which is expressed on B-cell lymphocytes and on the majority of B-cell lymphoid malignancies. Herein we report the conjugate of rituximab with the near-infrared (NIR) fluorophore Cy5.5 (RI-Cy5.5) as a tool for in vitro, in vivo, and ex vivo NIR time-domain (TD) optical imaging. In vitro, RI-Cy5.5 retained biologic activity and led to elevated cell-associated fluorescence on tumor cells. In vivo, TD optical imaging analysis of RI-Cy5.5 injected into lymphoma-bearing mice revealed a slow tumor uptake and a specific long-lasting persistence of the probe within the tumor. Biodistribution studies after intraperitoneal and endovenous administration were undertaken to evaluate differences in the tumor uptake. RI Cy5.5 concentration in the organs after intraperitoneal injection was not as high as after endovenous injection. Ex vivo analysis of biologic tissues and organs by both TD optical imaging and immunohistochemistry confirmed the probe distribution, as demonstrated by imaging experiment in vivo, showing that RI Cy5.5 selectively accumulated in the tumor tissue and major excretion organs. In summary, the study indicates that NIR TD optical imaging is a powerful tool for rituximab-targeting investigation, furthering understanding of its administration outcome in lymphoma treatment. PMID- 19123998 TI - Molecular imaging provides novel insights on estrogen receptor activity in mouse brain. AB - Estrogen receptors have long been known to be expressed in several brain areas in addition to those directly involved in the control of reproductive functions. Investigations in humans and in animal models suggest a strong influence of estrogens on limbic and motor functions, yet the complexity and heterogeneity of neural tissue have limited our approaches to the full understanding of estrogen activity in the central nervous system. The aim of this study was to examine the transcriptional activity of estrogen receptors in the brain of male and female mice. Exploiting the ERE-Luc reporter mouse, we set up a novel, bioluminescence based technique to study brain estrogen receptor transcriptional activity. Here we show, for the first time, that estrogen receptors are similarly active in male and female brains and that the estrous cycle affects estrogen receptor activity in regions of the central nervous system not known to be associated with reproductive functions. Because of its reproducibility and sensitivity, this novel bioluminescence application stands as a candidate as an innovative methodology for the study and development of drugs targeting brain estrogen receptors. PMID- 19123999 TI - Specificity analysis of lectins and antibodies using remodeled glycoproteins. AB - Due to their ability to bind specifically to certain carbohydrate sequences, lectins are a frequently used tool in cytology, histology, and glycan analysis but also offer new options for drug targeting and drug delivery systems. For these and other potential applications, it is necessary to be certain as to the carbohydrate structures interacting with the lectin. Therefore, we used glycoproteins remodeled with glycosyltransferases and glycosidases for testing specificities of lectins from Aleuria aurantia (AAL), Erythrina cristagalli (ECL), Griffonia simplicifolia (GSL I-B(4)), Helix pomatia agglutinin (HPA), Lens culinaris (LCA), Lotus tetragonolobus (LTA), peanut (Arachis hypogaeae) (PNA), Ricinus communis (RCA I), Sambucus nigra (SNA), Vicia villosa (VVA), and wheat germ (Triticum vulgaris) (WGA) as well as reactivities of anti-carbohydrate antibodies (anti-bee venom, anti-horseradish peroxidase [anti-HRP], and anti Lewis(x)). After enzymatic remodeling, the resulting neoglycoforms display defined carbohydrate sequences and can be used, when spotted on nitrocellulose or in enzyme-linked lectinosorbent assays, to identify the sugar moieties bound by the lectins. Transferrin with its two biantennary complex N-glycans was used as scaffold for gaining diverse N-glycosidic structures, whereas fetuin was modified using glycosidases to test the specificities of lectins toward both N- and O glycans. In addition, alpha(1)-acid glycoprotein and Schistosoma mansoni egg extract were chosen as controls for lectin interactions with fucosylated glycans (Lewis(x) and core alpha1,3-fucose). Our data complement and expand the existing knowledge about the binding specificity of a range of commercially available lectins. PMID- 19124000 TI - Interference of sugars in the Coomassie Blue G dye binding assay of proteins. AB - The presence of sugars causes significant deviation from the actual absorbance of proteins in the Bradford protein assay. In these studies, polysaccharides and disaccharides at milligram levels mimicked proteins in microgram equivalents. Monosaccharides, which individually did not show any absorbance, interfered significantly by sequestering the dye species. The studies demonstrated that in a mixture of sugars and proteins, sugar interference was much higher than expected from sugar molecules' individual contribution. Estimated protein values were increased 2 to 4 times after precipitation from fungal culture broths. Thus, in carbohydrate-rich samples, protein concentrations should be ascertained by precipitation from crude extracts and resolubilization in a noninterfering buffer. PMID- 19124001 TI - Transcription of a "silent" cyanobacterial psbA gene is induced by microaerobic conditions. AB - Cyanobacteria, contrary to higher plants, have a small psbA gene family encoding the reaction centre D1 protein subunit of photosystem II, the first macromolecular pigment-protein complex of the photosynthetic electron transport chain. Modulation of expression of multiple psbA genes in the family allows cyanobacteria to adapt to changing environmental conditions. To date, two different strategies for regulation of the psbA genes have emerged. One, characterized in Synechocystis PCC6803 and Gloeobacter violaceus PCC7421 involves the increased expression of one type of D1 protein to cope with the increased rate of damage. The other strategy, in Synechococcus PCC7942 and Anabaena PCC7120, is to replace the existing D1 with a new D1 form for the duration of the stress. However, most of the psbA gene families characterized to date contain also a divergent, apparently silent psbA gene of unknown function. This gene, present in Synechocystis, Anabaena and Thermosynechococcus elongatus BP-1 was not induced by any stress condition applied so far. Our data shows a reversible induction of the divergent psbA gene during the onset of argon-induced microaerobic conditions in Synechocystis, Anabaena and Thermosynechococcus elongatus. The unitary functional response of three unrelated cyanobacterial species, namely the induction of the expression of the divergent psbA gene as a reaction to the same environmental cue, indicates that these genes and the protein they encode are part of a specific cellular response to microaerobic conditions. There are no specific primary structure similarities between the different microaerobic inducible D1 forms, designated as D1'. Only three amino acid residues are consistently conserved in D1'. These modifications are: G80 to A, F158 to L and T286 to L. In silico mutation of the published D1 structure from Thermosynechococcus did not reveal major modifications. The point by point effects of the mutations on the local environment of the PSII structure are also discussed. PMID- 19124002 TI - Wild-type AIRE cooperates with p63 in HLA class II expression of medullary thymic stromal cells. AB - During T cell development in the thymus, autoreactive T cells are deleted through a mechanism that is actively supported by medullary epithelial cells. These epithelial cells possess particular transcription factors including autoimmune regulator (AIRE), which is responsible for regulating expression of self antigens, as well as p63, a p53-like molecule. Here we present evidence suggesting interaction of AIRE with p63 through a SAND domain and a transactivation domain, respectively. Interestingly an AIRE molecule with a mutated SAND domain of G228W, whose genetic alteration is inherited in an autosomal dominant manner, could not establish a complex with p63 as indicated by immunoprecipitation and molecular modeling analyses. Further in vitro study indicated that the G228W mutation led to downregulation of the transcription levels of CIITA and, accordingly, the cell surface expression of HLA class II molecules in thymic epithelial cells with p63. This indicates novel involvement of AIRE and p63 in the regulation of HLA class II, and suggests that defects in the AIRE-p63 interaction may lead to malfunction of HLA-based selection of self reactive helper CD4(+) T cells in the thymus. PMID- 19124003 TI - Hematopoiesis-dependent expression of CD44 in murine hepatic progenitor cells. AB - The fetal liver serves as the predominant hematopoietic organ until birth. However, the mechanisms underlying this link between hematopoiesis and hepatogenesis are unclear. Previously, we reported the isolation of a monoclonal antibody (anti-Liv8) that specifically recognizes an antigen (Liv8) present in murine fetal livers at embryonic day 11.5 (E11.5). Liv8 is a cell surface molecule expressed by hematopoietic cells in both fetal liver and adult mouse bone marrow. Here, we report that Liv8 is also transiently expressed by hepatoblasts at E11.5. Using protein purification and mass spectrometry, we have identified Liv8 as the CD44 protein. Interestingly, the expression of Liv8/CD44 in fetal liver was completely lost in AML1(-/-) murine embryos, which lack definitive hematopoiesis. These results show that hepatoblasts change from Liv8/CD44-negative to Liv8/CD44-positive status in a hematopoiesis-dependent manner by E11.5, and indicate that Liv8/CD44 expression is an important link between hematopoiesis and hepatogenesis during fetal liver development. PMID- 19124004 TI - CMTM5-v1 induces apoptosis in cervical carcinoma cells. AB - CMTM5 (CKLF-like MARVEL transmembrane domain-containing member 5) exhibits tumor inhibition activity with frequent epigenetic inactivation in various tumor cell lines including cervical carcinoma (CC) cells. In this paper, we examined the function of CMTM5-v1 (the primary RNA splicing form) in both HeLa and SiHa cells. Overexpression of CMTM5-v1 in both cells can induce apoptosis, but the effects are more obvious in SiHa than that in HeLa. In SiHa cells, restoration of CMTM5 v1 caused disruption of mitochondrial transmembrane potential, release of cytochrome c, activation of caspase3 and cleavage of PARP. General caspase inhibitor almost prevented apoptosis of SiHa cells, suggesting that CMTM5-v1 induces apoptosis mainly through caspase-dependent pathway. These findings verify that CMTM5-v1 inhibits the growth of CC cell lines via inducing apoptosis. PMID- 19124005 TI - Galectin-3 regulates apoptosis and doxorubicin chemoresistance in papillary thyroid cancer cells. AB - A subset of patients with papillary thyroid cancer (PTC) present with aggressive disease that is refractory to conventional treatment. Novel therapies are needed to treat this group of patients. Galectin-3 (Gal-3) is a beta-galactoside-binding protein with anti-apoptotic activity. Over 30 studies in the last 3 years have reported that Gal-3 is highly expressed in PTC relative to normal thyrocytes. In this study, we show that Gal-3 silencing with RNA interference stimulates apoptosis, while Gal-3 overexpression protects against both TRAIL- and doxorubicin-induced apoptosis in PTC cells. The anti-apoptotic activity and chemoresistance related to Gal-3 function can be partially reversed through the inhibition of the PI3K-Akt pathway, suggesting that Gal-3 acts, at least in part, on the PI3K-Akt axis. These observations support further evaluation of Gal-3 as a potential therapeutic target in patients with aggressive PTC. PMID- 19124006 TI - HSV-2 induces TLRs and NF-kappaB-dependent cytokines in cervical epithelial cells. AB - Herpes simplex virus type 2 (HSV-2) is one of the most common sexually transmitted pathogens worldwide. The host immune response induced by viral infection is cell-type specific. Little is known about the innate immune response to this virus in its natural host cells. In this study, we established an in vitro HSV-2 infection model with human cervical epithelial (HCE) cells. The viral infection was sufficient to induce expression of Toll-like receptors (TLRs), and Western blot and reporter assays suggest that HSV-2 infection leads to dramatic activation of the NF-kappaB signaling pathway. More importantly, our data provide direct evidence that the activation of NF-kappaB is required for the production of both IL-6 and IFN-beta induced by HSV-2 in HCE cells. Taken together, our results suggest the potential contributions of TLRs and a critical role of NF kappaB in the innate immune response to HSV-2 in HCE cells. PMID- 19124007 TI - Hyaluronan dynamics during retinal development. AB - We have already shown the regulated expressions of hyaluronan binding molecules including versican, SPACR, and SPACRCAN, during retinal development. Here we analyzed the profiles of hyaluronan during these periods. Hyaluronan and hyaluronan synthase (HAS) expressions in chicken retinas at different ages were assessed by slot blot and real-time PCR, respectively. The molecular size of hyaluronan was determined using a Superose-6 column, and histochemistry was used to localize hyaluronan expression. Hyaluronan degradation activity was measured after adding hyaluronan to retinal extracts. In the embryonic retina, hyaluronan was mainly present in the inner plexiform and photoreceptor layers. Expression in the inner plexiform layer decreased with development and aging while expression in the photoreceptor layer remained constant. The molecular size of hyaluronan decreased from embryonic day 12 (E12) to postnatal day 1 (P1) with increasing retinal hyaluronan degradation activity. In adult retinas, hyaluronan was only present in the photoreceptor layer, and its molecular size was the highest. HAS-2 and HAS-3 expressions increased from E15 to P1, but both were reduced in adulthood. Thus hyaluronan was specifically detected where versican, SPACR, and SPACRCAN were previously demonstrated, and the synthesis and degradation of hyaluronan are regulated. Hyaluronan together with versican, SPACR, and SPACRCAN are closely related during retinal development, and may be important for retinal physiology. PMID- 19124008 TI - Radial glia-like cells persist in the adult rat brain. AB - During development, radial glia cells contribute to neuronal migration and neurogenesis, and differentiate into astrocytes by the end of the developmental period. Recently, it was demonstrated that during development, radial glia cells, in addition to their role in migration, also give rise to neuroblasts. Furthermore, radial glial cells remain in the adult brain as adult neural stem cells (NSC) in the subventricular zone (SVZ) around the lateral ventricles (LVs), and generate new neurons continuously throughout adulthood. In this study, we used immunohistochemical and morphological methods to investigate the presence of radial glia-like cells around the LVs during the postnatal development period until adulthood in rats. In all ages of rats studied, we identified cells with morphological and immunocytochemical features that are similar to the radial glia cells found in the embryonic brain. Similarly to the radial glia, these cells express nestin and vimentin, and have a radial morphology, extending perpendicularly as processes from the ventricle wall. These cells also express GFAP, GLAST, and Pax6, and proliferate. In the brains of adult rats, we identified cells with relatively long processes (up to 600 mum) in close apposition with migrating neuroblasts. Our results showed that the radial glia like cells present in the adult rat brain share several morphological and functional characteristics with the embryonic radial glia. We suggest that the embryonic radial glia cells located around the LV walls do not complete their transformation into astrocytes, but rather persist in adulthood. PMID- 19124009 TI - Lack of association between the polymorphisms of beta-site APP-cleaving enzyme 2 (BACE2) 5'-flanking region and sporadic Alzheimer's disease. AB - Amyloid beta-peptide (A beta) plays a central role in the pathogenesis of Alzheimer's disease (AD). A beta is produced by sequential cleavage of the amyloid precursor protein (APP) by two enzymes referred to as beta- and gamma secretase. beta-secretase is of more importance, as it catalyses the rate limiting step in the production of A beta. Although beta-site APP-cleaving enzyme 1 (BACE1) is known to cleave APP at the beta-secretase site as required for the generation of A beta, the role of its homologue BACE2 is controversial. For seeking the correlation of the BACE2 promoter with sporadic AD (SAD), we performed a case-control study in a Chinese Han population. In the study, we sequenced the 2641 bp fragment of the 5'-flanking region of BACE2 gene and found three polymorphisms which are -320C/- (rs11316732), -1541A/T (rs9975138) and 1904C/T (rs28656880). Definitive genotyping these markers and apolipoprotein E (APOE) polymorphism were surveyed using restriction enzyme digestion and direct sequencing in 359 SAD patients and 334 controls. We failed to find any association between these three polymorphisms and SAD even after statistical adjustment for age, gender and APOE epsilon 4 status. Our data do not support that there is a linkage between the 5'-flanking region polymorphisms of BACE2 and SAD in the Chinese Han population. PMID- 19124010 TI - Effects of acute and repeated restraint stress on GABA efflux in the rat basolateral and central amygdala. AB - Stress can precipitate onset of multiple mood-related disorders, including depression. Examination of the neural basis of this phenomenon has highlighted the amygdala as a key component. Alterations in amygdalar activity and structure accompany various mood-related disorders, and interestingly, amygdalar morphology and behavior can be altered in animals subjected to repeated stress. Gamma aminobutyric acid (GABA) transmission in the amygdala represents an important means by which information flow, activity, and function can be controlled; therefore, we determined the effects of acute and repeated restraint stress (RRS) on GABA efflux in the basolateral and central amygdalar complexes. In vivo microdialysis revealed that acute restraint stress increased GABA efflux in the basolateral amygdala, whereas central amygdala efflux remained unchanged. Animals subjected to prior repeated stress displayed no acute stress-mediated increases in GABA efflux in the basolateral amygdala, an event accompanied by no changes in basal GABA concentrations. Conversely, repeated restraint stress had no effect on GABA efflux or basal GABA levels in the CeA. Collectively, these data demonstrate that acute stress elicits unique and region-specific increases in GABA efflux in the rat amygdala, and that prior repeated stress differentially modifies this response. PMID- 19124011 TI - Perinatal exposure to PTU decreases expression of Arc, Homer 1, Egr 1 and Kcna 1 in the rat cerebral cortex and hippocampus. AB - Environmental chemicals have a potential impact on neuronal development and children's health. The current developmental neurotoxicity (DNT) guideline studies to assess their underlying risk are costly and time-consuming; therefore the more efficient protocol for DNT test is needed. Hypothyroidism in rats induced by perinatal exposure to propylthiouracil (PTU), a thyroid hormone synthesis inhibitor, offers an advantageous model of developmental neurotoxicity (DNT). Understanding the associated alterations in gene expression in brain is a key to elucidate mechanisms and find appropriate molecular markers. The purpose of the present study was to identify PTU treatment-affected transcriptomes in the rat cerebral cortex and the hippocampus using DNA microarrays, and to specify candidate genes linked to DNT. We used an approximately 9000 probe microarray to examine differentially expressed genes between PTU-dosed and vehicle-dosed rats at postnatal days 4, 14, 22 and 70. Expression of immediate early genes (IEGs) such as activity-regulated cytoskeleton-associated protein (Arc), Homer 1, early growth response 1 (Egr 1), myelin-associated genes such as myelin-associated oligodendrocytic basic protein (MOBP), myelin basic protein (MBP) and proteolipid protein (PLP) and Kcna1 was apparently affected by perinatal administration of PTU. The results suggest that the alterations may be responsible for the detrimental effects caused by PTU treatment on the nervous system. PMID- 19124012 TI - Cerebral metabolic effects of acetyl-l-carnitine in rats during aging. AB - The aim of the present study was to investigate the neuronal structures that mediate the antiaging properties of acetyl-l-carnitine (ALCAR). The regional cerebral metabolic rates for glucose (rCMRglc) have been determined with the quantitative autoradiographic [(14)C]2-deoxyglucose procedure at different times after i.v. administration of saline or ALCAR 500 mg/kg to naive, non pretreated 3 , 12- and 24-month-old rats and to 24-month-old rats pretreated with ALCAR (100 mg/kg/day, for 3 months). rCMRglc increased maximally at 30 min after ALCAR in 3 , 12- and 24-month old rats (14, 15 and 15 areas affected, 19, 24 and 22% mean increments). Peak metabolic activations occurred with similar magnitude in motor, visual, limbic and thalamic areas in all age rats and with larger magnitude in hippocampal and thalamic areas in aged rats. Cerebral metabolic activations subsided by 60 min after ALCAR in 3-month rats (3 brain regions affected, 4% decrease) and persisted by that time in 12- and 24-month-old rats (14 and 12 regions affected, 15 and 20% increases). Cerebral activations were enhanced in aged rats after chronic treatment with ALCAR (24 brain regions affected, 20% mean increase). Hence, during aging, metabolic responsivity to ALCAR is maintained in most brain areas and increased in limbic and thalamic regions. Increased responsivity to ALCAR may result from undetermined pharmacokinetic factors and/or from a higher sensitivity and contribute to the aging reversal properties of ALCAR. PMID- 19124013 TI - Method specific second-trimester reference intervals for thyroid-stimulating hormone and free thyroxine. AB - OBJECTIVE: To determine second trimester reference intervals for TSH and FT4. DESIGN: Samples from 3102 subjects were tested for TPO and Tg antibodies. METHODS: Elecsys E170 reference intervals for TSH and FT4 were determined using antibody-negative samples. RESULTS: Second trimester reference intervals for TSH and FT4 were 0.18-4.07 mIU/L and 9.5-15.8 pmol/L, respectively. The Elecsys E170 TSH results were positively biased compared to ARCHITECT i2000(SR) results for these same samples. CONCLUSIONS: Method-specific reference intervals are required for TSH and FT4. PMID- 19124014 TI - Testis cord differentiation after the sex determination stage is independent of Sox9 but fails in the combined absence of Sox9 and Sox8. AB - Sox9 and Sox8 are transcription factors expressed in embryonic and postnatal Sertoli cells of the mouse testis. Sox9 inactivation prior to the sex determination stage leads to complete XY sex reversal. In contrast, there is normal embryonic testis development in Sox8 mutants which are initially fertile, but later develop progressive seminiferous tubule failure and infertility. To determine whether Sox9 is required for testis development after the initial steps of sex determination, we crossed Sox9(flox) mice with an AMH-Cre transgenic line thereby completely deleting Sox9 in Sertoli cells by E14.0. Conditional Sox9 null mutants show normal embryonic testis development and are initially fertile, but, like Sox8(-/-) mutants, become sterile from dysfunctional spermatogenesis at about 5 months. To see whether Sox8 may compensate for the absence of Sox9 during embryonic testis differentiation, we generated a Sox9 conditional knockout on a Sox8 mutant background. In the double mutants, differentiation of testis cords into seminiferous testis tubules ceases after P6 in the absence of one Sox8 allele, and after P0 in the absence of both Sox8 alleles, leading to complete primary infertility. Sox9,Sox8 double nullizygous testes show upregulation of early ovary-specific markers and downregulation of Sertoli intercellular junctions at E15.5. Their very low Amh levels still cause complete regression of the Mullerian duct but with reduced penetrance. This study shows that testis cord differentiation is independent of Sox9, and that concerted Sox9 and Sox8 function in post E14.0 Sertoli cells is essential for the maintenance of testicular function. PMID- 19124015 TI - Expression of Akt and Mdm2 in human esophageal squamous cell carcinoma. AB - The Akt-Mdm2 pathway plays an important role in carcinogenesis in a variety of malignant tumors. However, the Akt-Mdm2 pathway in esophageal squamous cell carcinoma (ESCC) has not been fully studied. We investigated the proteins and mRNA expression of Akt and Mdm2 to elucidate the roles of these proteins in ESCC. We also examined the effect of Akt knockdown on Mdm2 expression in ESCC cells. ESCC tissue samples were obtained from 23 individuals who underwent surgical resection with no preoperative treatment. Akt1-3 and Mdm2 gene and protein expression were analyzed. The effect of siRNA-mediated Akt knockdown on Mdm2 expression was also studied, using ESCC cell lines. Akt1 and Mdm2 immunoreactivity was detected in 77.8 and 66.7% of tumor specimen from ESCC patients, respectively. Akt1 and Mdm2 mRNA expressions were correlated and significantly elevated in tumor tissue (p<0.0001 and p<0.05, respectively). The siRNA-targeted reduction of each Akt isoform reduced Mdm2 protein expression. The overexpression of Akt1 and Mdm2 may be related to esophageal carcinogenesis. Furthermore, Akt expression regulates Mdm2 expression, which may in turn regulate the function of wild-type p53. These results may provide the basis for future preventative or clinical therapies for esophageal cancer. PMID- 19124016 TI - PRMT1 mediated methylation of TAF15 is required for its positive gene regulatory function. AB - TAF15 (formerly TAF(II)68) is a nuclear RNA-binding protein that is associated with a distinct population of TFIID and RNA polymerase II complexes. TAF15 harbours an N-terminal activation domain, an RNA recognition motif (RRM) and many Arg-Gly-Gly (RGG) repeats at its C-terminal end. The N-terminus of TAF15 serves as an essential transforming domain in the fusion oncoprotein created by chromosomal translocation in certain human chondrosarcomas. Post-transcriptional modifications (PTMs) of proteins are known to regulate their activity, however, nothing is known on how PTMs affect TAF15 function. Here we demonstrate that endogenous human TAF15 is methylated in vivo at its numerous RGG repeats. Furthermore, we identify protein arginine N-methyltransferase 1 (PRMT1) as a TAF15 interactor and the major PRMT responsible for its methylation. In addition, the RGG repeat-containing C-terminus of TAF15 is responsible for the shuttling between the nucleus and the cytoplasm and the methylation of RGG repeats affects the subcellular localization of TAF15. The methylation of TAF15 by PRMT1 is required for the ability of TAF15 to positively regulate the expression of the studied endogenous TAF15-target genes. Our findings demonstrate that arginine methylation of TAF15 by PRMT1 is a crucial event determining its proper localization and gene regulatory function. PMID- 19124017 TI - Synemin isoforms during mouse development: multiplicity of partners in vascular and neuronal systems. AB - The intermediate filament (IF) synemin gene encodes three IF proteins (H 180, M 150, L 41 kDa isoforms) with overlapping distributions. In the present study we analysed the mRNA and protein expression of each isoform in developing mouse embryos. Synemin M mRNA was present as early as E5 with vimentin and nestin. Synemin H was found later at E9 in the nervous system and mesodermic derivatives concomitantly with angiogenesis, somitogenesis and the migration of neural crest cells. Synemin L appeared later in neurons at E15. Furthermore, the synemin isoforms required different IF partners depending on the cell type to form filamentous structures. In endothelial cells, synemin H/M were found associated with vimentin and were absent in vimentin-null mice. In neurons of the peripheral nervous system of E15 embryos, synemin H/M or L were co-expressed with neurofilament, peripherin and internexin. In adult mice, our data support the existence of different subpopulations of neurons within the dorsal root ganglia: one composed of small neurons containing synemin H/M and peripherin, and another composed of large neurons containing synemin L and neurofilaments. Axons devoid of neurofilaments from mutant mice (NFHLacZ) showed an absence of the L isoform but contained H/M isoforms with peripherin. PMID- 19124018 TI - Short hypoxia could attenuate the adverse effects of hyperhomocysteinemia on the developing rat brain by inducing neurogenesis. AB - Gestational deficiency in methyl donors such as folate and vitamin B12 impairs homocysteine metabolism and can alter brain development in the progeny. Since short hypoxia has been shown to be neuroprotective in preconditioning studies, we aimed to investigate the effects of brief, non-lesioning neonatal hypoxia (100% N2 for 5 min) on the developing brain of rats born to dams fed either a standard diet or a diet lacking vitamins B12, B2, folate and choline until offspring's weaning. While having no influence on brain accumulation of homocysteine and concomitant apoptosis in 21-day-old deficient pups, exposure to hypoxia reduced morphological injury of the hippocampal CA1 layer. It also markedly stimulated the incorporation of bromodeoxyuridine (BrdU) in permissive areas such as the subventricular zone and the hippocampus followed by the migration of new neurons. Scores in a locomotor coordination test (days 19-21) and learning and memory behavior in the eight-arm maze (days 80-84) were found to be significantly improved in rats exposed to hypoxia in addition to the deficient diet. Therefore, by stimulating neurogenesis in rat pups, brief neonatal hypoxia appeared to attenuate the long-term effects of early exposure to a deficiency in nutritional determinants of hyperhomocysteinemia. PMID- 19124019 TI - Recurrent apnea induces neuronal apoptosis in the guinea pig forebrain. AB - Obstructive sleep apnea (OSA) and sleep-disordered breathing (SDB) can result in impaired cognition and mental acuity, and the generation of mood disorders, including depression. However, the mechanisms of neuronal damage for these complications have not been elucidated. Accordingly, using immunohistochemical technique with monoclonal antibody against single-stranded DNA, we examined the morphological effects of chronic recurrent apnea on neurons in the hippocampus and related forebrain sites in guinea pigs. Our results show that a large number of neurons labeled by anti-ssDNA antibody were present in the cingulate, insular and frontal cortices, the hippocampus and the amygdala in conjunction with periods of recurrent apnea. However, no labeling was observed in comparable regions of the brain in control guinea pigs. In the cortices of experimental animals, labeled neurons were detected mainly in the superficial layers (II-III) in the frontal, insular and cingulate cortex. In the hippocampus, most labeled neurons were located in the CA1 region, in which most of stained neurons were observed in strata pyramidal, while only a few positive neurons were located in the strata radiatum and the strata oriens. In addition, a large number of labeled neurons were also detected in the central nucleus of amygdala in the guinea pigs underwent recurrent periods of apnea. The present data indicate that recurrent apnea results in cell death in the hippocampus and related forebrain regions via mechanisms of apoptosis, which may represent the basis for the clinical complications of obstructive sleep apnea and sleep-disordered breathing. PMID- 19124020 TI - Selective leishmanicidal effect of 1,3,4-thiadiazole derivatives and possible mechanism of action against Leishmania species. AB - With the aim of determining selectivity and the possible target(s) of nitroheteroaryl-1,3,4-thiadiazoles considered as possible leads for the development of anti-leishmanial agents, we studied 5-nitroimidazole, 5-nitrofuran and 5-nitrothiophene analogs of N-substituted-piperazinyl-1,3,4-thiadiazoles. We investigated 21 representative compounds 1-3(a-g) for the following properties: selectivity and efficiency against different Leishmania wild type species and intracellular parasite, toxicity against host cells and inhibition of topoisomerases I and II. Our results indicate that the nitroimidazole analogs 1a and 1f, and nitrofuran derivatives 2a, 2b, 2c, 2f, and 2g exhibited low toxicity against the host cells (IC(50)> or =80 microM), but high selectivity against intracellular amastigotes (selectivity index>12). Leishmania topoisomerases revealed impressive sensitivity to the agents (%inhibition >50 at IC(50) doses of compounds against Leishmania). Our findings showed that at least part of leishmaniacidal effect of the compounds could be attributed to disruption DNA relaxed activities of topoisomerases I and II, and cleavable-complex formation. PMID- 19124021 TI - Multiplex real-time PCR detection of P. falciparum, P. vivax and P. malariae in human blood samples. AB - Two duplex real-time PCR assays were developed to diagnose three human parasites: Plasmodium falciparum, Plasmodium vivax and Plasmodium malariae. TaqMan duplex real-time PCR was evaluated in 263 blood samples of suspected malaria patients by comparing results against those obtained with microscopy and nested PCR. Compared with nested PCR, duplex real-time PCR assays showed 100% sensitivity and specificity. Duplex real-time PCR detected all mixtures of P. falciparum and P. vivax DNA, except at threshold detection limits for both parasites in which P. vivax was not amplified. Threshold detection limits of real-time PCR were 3.1, 0.3 and 0.8 parasites per microlitre of blood for P. falciparum, P. vivax and P. malariae, respectively. Duplex real-time PCR allows the detection of malarial cases, including mixed species infection, it simplifies analysis and reduces cost. Thus, this protocol may prove invaluable for use in the diagnosis of human infection, trial treatments and epidemiologic studies in which high-throughput analyses are often required. PMID- 19124022 TI - Electronic clinical challenges and images in GI. Image 2. PMID- 19124023 TI - Burden of digestive diseases in the United States part I: overall and upper gastrointestinal diseases. PMID- 19124024 TI - Dog's gaze at its owner increases owner's urinary oxytocin during social interaction. AB - Oxytocin (OT) has been shown to play an important role in social bonding in animals. However, it is unclear whether OT is related to inter-species social bonding. In this study, to examine the possibility that urinary OT concentrations of owners were increased by their "dog's gaze", perhaps representing social attachment to their owners, we measured urinary OT concentrations of owners before and after interaction with their dogs. Dog owners interacted with their dogs as usual for 30 min (interaction experiment) or were instructed not to look at their dogs directly (control experiment). We observed the behaviors of owners and their dogs during the experiments, and measured OT concentrations by radioimmunoassay in urine samples from the owners collected just before and 20 min after interaction with their dogs. Using a cluster analysis, owners could be divided into two groups: one received a longer duration of gaze from their dogs and reported a higher degree of relationship with their dogs (LG); the other received a shorter duration of gaze and reported a lower degree of relationship (SG). Urinary OT was higher in LG than SG after usual interaction with their dogs, but not in the control experiment. In the interaction experiment, a high correlation was found in LG between the frequency of behavioral exchanges initiated by the dog's gaze and the increase in urinary OT. We conclude that interactions with dogs, especially those initiated by the dog's gaze, can increase the urinary OT concentrations of their owners as a manifestation of attachment behavior. PMID- 19124026 TI - Insight on the molecular envelope of lipid-bound apolipoprotein E from electron paramagnetic resonance spectroscopy. AB - Although a high-resolution X-ray structure for the N-terminal domain of apolipoprotein E (apoE) in the lipid-free state has been solved, our knowledge of the structure of full-length apoE in a lipid-bound state is limited to an X-ray model fitting a molecular envelope at 10-A resolution. To add molecular detail to the molecular envelope, we used cysteine mutagenesis to incorporate spin labels for analysis with electron paramagnetic resonance (EPR) spectroscopy. Twelve cysteine residues were introduced singly and in pairs at unique locations throughout apoE4 and labeled with an EPR spin probe. The labeled apoE4 was combined with dipalmitoylphosphatidylcholine, the particles were purified, and spectra were determined for 24 combinations (single and double) of the cysteine mutants. Data on the conformation, mobility, distance, and surface exposure of regions revealed by the cysteine probes were modeled into the molecular envelope of apoE bound to dipalmitoylphosphatidylcholine that had been determined by X-ray analysis. This EPR model of apoE in a native lipid-bound state validates the structural model derived from X-ray analysis and provides additional insight into apoE structure-function relationships. PMID- 19124025 TI - Chronic estradiol treatment increases CA1 cell survival but does not improve visual or spatial recognition memory after global ischemia in middle-aged female rats. AB - Transient global ischemia induces selective, delayed neuronal death in the hippocampal CA1 and cognitive deficits. Physiological levels of 17beta-estradiol ameliorate ischemia-induced neuronal death and cognitive impairments in young animals. In view of concerns regarding hormone therapy in postmenopausal women, we investigated whether chronic estradiol treatment initiated 14 days prior to ischemia attenuates ischemia-induced CA1 cell loss and impairments in visual and spatial memory, in ovariohysterectomized (OVX), middle-aged (9-11 months) female rats. To determine whether the duration of hormone withdrawal affects the efficacy of estradiol treatment, hormone treatment was initiated immediately (0 week), 1 week, or 8 weeks after OVX. Age-matched, OVX and gonadally intact females were studied at each OVX interval. Ischemia was induced 1 week after animals were pretested on a variety of behavioral tasks. Global ischemia produced significant neuronal loss in the CA1 and impaired performance on visual and spatial recognition. Chronic estradiol modestly but significantly increased the number of surviving CA1 neurons in animals at all OVX durations. However, in contrast with previous results in young females, estradiol did not preserve visual or spatial memory performance in middle-aged females. All animals displayed normal locomotion, spontaneous alternation and social preference, indicating the absence of global behavioral impairments. Therefore, the neuroprotective effects of estradiol are different in middle-aged than in young rats. These findings highlight the importance of using older animals in studies assessing potential treatments for focal and global ischemia. PMID- 19124027 TI - Insights into the conformational variability and regulation of human Nek2 kinase. AB - The Nek family of serine/threonine kinases regulates centrosome and cilia function; in addition, several of its members are potential targets for drug discovery. Nek2 is dimeric, is cell cycle regulated and functions in the separation of centrosomes at G2/M. Here, we report the crystal structures of wild type human Nek2 kinase domain bound to ADP at 1.55-A resolution and T175A mutant in apo form as well as that bound to a non-hydrolyzable ATP analog. These show that regions of the Nek2 structure around the nucleotide-binding site can adopt several different but well-defined conformations. None of the conformations was the same as that observed for the previously reported inhibitor-bound structure, and the two nucleotides stabilized two conformations. The structures suggest mechanisms for the auto-inhibition of Nek2 that we have tested by mutagenesis. Comparison of the structures with Aurora-A and Cdk2 gives insight into the structural mechanism of Nek2 activation. The production of specific inhibitors that target individual kinases of the human genome is an urgent challenge in drug discovery, and Nek2 is especially promising as a cancer target. We not only identify potential challenges to the task of producing Nek2 inhibitors but also propose that the conformational variability provides an opportunity for the design of Nek2 selective inhibitors because one of the conformations may provide a unique target. PMID- 19124028 TI - Identification and functional analysis of a novel TRPC6 mutation associated with late onset familial focal segmental glomerulosclerosis in Chinese patients. AB - OBJECTIVE: Mutations in the TRPC6 gene are responsible for a late onset form of familial focal segmental glomerulosclerosis (FSGS). However, the role of TRPC6 variation in Chinese patients with late onset familial FSGS remains unclear. Here, we screened 31 Chinese pedigrees with late onset familial FSGS for changes in TRPC6 by DNA sequence analysis. METHODS: Genomic DNA was extracted from peripheral leukocytes. We PCR-amplified each of 13 exons of TRPC6 for sequence analysis. When a novel nucleotide change seemed likely to cause FSGS, we carried out an in vitro research to determine the effects of the mutation on TRPC6 function. HEK 293 cells were transfected stably with vectors containing mutant or wild type TRPC6 cDNA. We then compared the expression of mutant TRPC6 to wild type TRPC6 using Western blot. For the observation of the function of mutant TRPC6 channel compared with wild type TRPC6 channel, Intracellular Ca(2+) concentration was detected using fluorescent indicator Fluo-3 among different groups of cells upon stimulation with 1-oleoyl-2-acetyl sn-glycerol (OAG). RESULTS: All the 31 pedigrees with late onset familial FSGS were collected in our department from September 1997 to October 2007. A novel TRPC6 mutation (cytosine 2664 adenine resulting in Glutamine 889 Lysine substitution, Q889K) was identified in one of these pedigrees. Mutant TRPC6 (TRPC6(Q889K)) or wild type TRPC6 was stably expressed in HEK293 cells by Western blot. The mutant TRPC6 expression was a little increased without significant difference compared with wild type TRPC6 expression, whereas the intracellular Ca(2+) level in cells expressing mutant TRPC6 was significantly increased compared with that in the cells expressing wild TRPC6 upon stimulation. CONCLUSION: We identified a novel TRPC6 mutation Q889K associated with late onset FFSGS in Chinese pedigrees and this mutation was demonstrated to be "gain of function" by an in vitro functional research. PMID- 19124029 TI - Spatial attention and interhemispheric visuomotor integration in the absence of the corpus callosum. AB - In the lateralized simple reaction time (SRT) task with unimanual responses (Poffenberger paradigm), reaction times (RTs) are faster with ipsilateral (uncrossed) than with contralateral (crossed) response hand-target hemifield combinations. The difference between crossed and uncrossed responses (CUD) has typically been interpreted to reflect callosal transfer time. Indeed, acallosal subjects and split-brain subjects have longer CUDs than control subjects. However, a few recent studies have demonstrated that, contrary to classical findings, the CUD is also affected by non-anatomical factors. Here we show that the CUD is also affected by non-anatomical factors in patients with agenesis of the corpus callosum and complete commissurotomy where interhemispheric transfer must be subcallosal. We tested acallosal subject M.M. and split brain patient A.A. on a lateralized SRT task with their arms alternately uncrossed (natural arms position) or crossed (unnatural arms position) across blocks of trials. The results revealed a significant effect of arms crossing on the size and direction of the CUD as previously found in normal subjects [Mooshagian, E., Iacoboni, M., & Zaidel, E. (2008). The role of task history in simple reaction time to lateralized light flashes. Neuropsychologia, 46(2), 659-664]. This suggests that non-anatomical factors that modulate interhemispheric visuomotor integration may occur in absence of the corpus callosum. Anterior commissure and interhemispheric cortico-subcortical pathways are likely implicated in these effects. PMID- 19124030 TI - Recall of remote episodic memories can appear deficient because of a gist-based retrieval orientation. AB - Determining whether patients with amnesia can succeed in remembering their distant past has pivotal implications for theories of memory storage. However, various factors influence recall. We speculated that some patients with anterograde amnesia adopt a gist-based retrieval orientation for memories from all time periods, thereby exaggerating remote recall deficits. We tested whether an experimentally induced gist-based retrieval orientation could indeed hinder remote recall. Healthy individuals described photographs of complex scenes (e.g., of a cluttered desk) either with many words or few words (detail- or gist-based manipulation, respectively). They subsequently recalled autobiographical events and produced less episodic information after engaging the gist-based compared to the detail-based orientation. These results demonstrate the ease with which a gist-based orientation can produce apparent recall impairments. Deficits in remote episodic recall, and in future-event imagining, must thus be interpreted in light of habitual tendencies toward gist-based retrieval that some amnesic patients may exhibit. PMID- 19124031 TI - Attention effects on auditory scene analysis in children. AB - Auditory scene analysis begins in infancy, making it possible for the baby to distinguish its mother's voice from other noises in the environment. Despite the importance of this process for human behavior, the question of how perceptual sound organization develops during childhood is not well understood. The current study investigated the role of attention for perceiving sound streams in a group of school-aged children and young adults. We behaviorally determined the frequency separation at which a set of sounds was detected as one integrated or two separated streams and compared these measures with passively and actively obtained electrophysiological indices (mismatch negativity (MMN) and P3b) of the same sounds. In adults, there was a high degree of concordance between passive and active electrophysiological indices of stream segregation that matched with perception. In contrast, there was a large disparity in children. Active electrophysiological indices of streaming were concordant with behavioral measures of perception, whereas passive indices were not. In addition, children required larger frequency separations to perceive two streams compared to adults. Our results suggest that differences in stream segregation between children and adults reflect an under-development of basic auditory processing mechanisms, and indicate a developmental role of attention for shaping physiological responses that optimize processes engaged during passive audition. PMID- 19124033 TI - The role of selective attention in matching observed and executed actions. AB - Substantial evidence suggests that observed actions can engage their corresponding motor representations within the observer. It is currently believed that this process of observation-execution matching occurs relatively automatically, without the need for top-down control. In this study we tested the susceptibility of the observation-execution matching process to selective attention. We used a Go/NoGo paradigm to investigate the phenomenon of 'automatic imitation', in which participants are faster to initiate a hand movement that is congruent with a concurrently observed action, relative to one that is incongruent. First, we replicated previous findings of automatic imitation, and excluded the possibility that spatial compatibility effects might explain these results (Experiment 1). We then presented participants with the same goal directed actions while directing their attention to an imperative stimulus that spatially overlapped, but was distinct from, the observed actions (Experiment 2). Crucially, automatic imitation no longer occurred when participants directed their attention away from the displayed actions and towards the spatially overlapping stimulus. In a final experiment, we examined whether the automatic imitation of grasp persists when participants attend to an irrelevant feature of the observed action, such as whether it is performed by a left or right hand (Experiment 3). Here we found that automatic imitation is contingent on participants attending to the feature of the observed hand that was relevant to their responses. Together these findings demonstrate the importance of selective mechanisms in the filtering of task-irrelevant actions, and indicate a role for top-down control in limiting the motoric simulation of observed actions. PMID- 19124032 TI - Aging and cortical mechanisms of speech perception in noise. AB - Spoken language processing in noisy environments, a hallmark of the human brain, is subject to age-related decline, even when peripheral hearing might be intact. The present study examines the cortical cerebral hemodynamics (measured by fMRI) associated with such processing in the aging brain. Younger and older subjects identified single words in quiet and in two multi-talker babble noise conditions (SNR 20 and -5dB). Behaviorally, older and younger subjects did not show significant differences in the first two conditions but older adults performed less accurately in the SNR -5 condition. The fMRI results showed reduced activation in the auditory cortex but an increase in working memory and attention related cortical areas (prefrontal and precuneus regions) in older subjects, especially in the SNR -5 condition. Increased cortical activities in general cognitive regions were positively correlated with behavioral performance in older listeners, suggestive of a compensatory strategy. Furthermore, inter-regional correlation revealed that while younger subjects showed a more streamlined cortical network of auditory regions in response to spoken word processing in noise, older subjects showed a more diffused network involving frontal and ventral brain regions. These results are consistent with the decline-compensation hypothesis, suggestive of its applicability to the auditory domain. PMID- 19124034 TI - Knowing we know before we know: ERP correlates of initial feeling-of-knowing. AB - Subjects performed a rapid feeling-of-knowing task developed by (Reder, L. M., & Ritter, F. (1992). What determines initial feeling of knowing? Familiarity with question terms, not with the answer. Journal of Experimental Psychology: Learning, Memory, and Cognition, 18, 435-451), while event-related potentials (ERPs) were recorded to identify the time course of "feeling-of-knowing" signals. Subjects were shown a series of math problems, some of which were repeated multiple times during the course of the experiment, and subjects had to rapidly decide whether the answer to a given problem could be quickly retrieved from memory (retrieval trials) or had to be calculated on scrap paper (calculate trials). Behavioral results replicated the 1992 study, showing that subjects can estimate whether the answer is known much faster than the answer can be retrieved. ERPs time-locked to the onset of the math problem showed that accurate retrieval trials were associated with greater positivity for an early frontal P2 component (epoched from 180 to 280ms) and a frontal-central P3 component (epoched from 300 to 550ms). Moreover, this feeling-of-knowing signal was not found for subjects who never obtained a successful on-time retrieval. We interpret these findings as suggesting that initial feeling-of-knowing relies on a rapid assessment of the "perceptual fluency" with which the stimulus is processed. If a stimulus is deemed sufficiently familiar, the activation level of an internal problem representation is used to arrive at a decision of whether to search for the answer or to calculate it. PMID- 19124035 TI - Adaptive changes of inner retina function in response to sustained pattern stimulation. AB - We have characterized adaptive changes of inner retina function in response to sustained pattern stimulation in 32 normal subjects with an age range 23-77 years by measuring changes of the pattern electroretinogram (PERG) as a function of time. Contrast-reversal stimuli had square-wave profile in space and time, with peak spatial and temporal frequency and high contrast to maximize response amplitude. The PERG signal was sampled over 5min with a resolution of 15s. PERG signals were non-stationary, resulting in either progressive amplitude decline or even enhancement to a plateau, with a time course that could be well described by an exponential function with a time constant of 1-2min. Higher initial amplitudes were generally associated with amplitude decline, and lower initial amplitudes with enhancement. The delta amplitude (plateau minus initial) was a linear function of the initial amplitude. The magnitude of delta decreased with decreasing initial amplitude and inverted its sign for initial amplitudes about 1/3 lower than the maximum initial amplitude measured, but still about 3-4 times larger than the noise. Amplitude decline was generally associated with phase lag, whereas amplitude enhancement was associated with phase advance. Altogether, PERG generators appear to slowly adjust their gain in order to keep their sustained activity at an intermediate level that is rather independent of the level of activity at stimulus onset. This behavior is reminiscent of a buffering mechanism, where glial cells may play a primary role. An energy-budget model of neural-vascular-glial interaction is provided together with an equivalent electrical circuit that accounts for the results. PMID- 19124036 TI - On the relation between dichoptic masking and binocular rivalry. AB - When our two eyes view incompatible images, the brain invokes suppressive processes to inhibit one image, and favor the other. Two phenomena are typically observed: dichoptic masking (reduced sensitivity to one image) for brief presentations, and binocular rivalry (alternation between the two images), over longer exposures. However, it is not clear if these two phenomena arise from a common suppressive process. We investigated this by measuring both threshold elevation in simultaneous dichoptic masking and mean percept durations in rivalry, whilst varying relative stimulus orientation. Masking and rivalry showed significant correlations, such that strong masking was associated with long dominance durations. A second experiment suggested that individual differences across both measures are also correlated. These findings are consistent with varying the magnitude of interocular suppression in computational models of both rivalry and masking, and imply the existence of a common suppressive process. Since dichoptic masking has been localised to the monocular neurons of V1, this is a plausible first stage of binocular rivalry. PMID- 19124037 TI - Identification of everyday objects on the basis of kinetic contours. AB - Using kinetic contours derived from everyday objects, we investigated how motion affects object identification. In order not to be distinguishable when static, kinetic contours were made from random dot displays consisting of two regions, inside and outside the object contour. In Experiment 1, the dots were moving in only one of two regions. The objects were identified nearly equally well as soon as the dots either in the figure or in the background started to move. RTs decreased with increasing motion coherence levels and were shorter for complex, less compact objects than for simple, more compact objects. In Experiment 2, objects could be identified when the dots were moving both in the figure and in the background with speed and direction differences between the two. A linear increase in either the speed difference or the direction difference caused a linear decrease in RT for correct identification. In addition, the combination of speed and motion differences appeared to be super-additive. PMID- 19124038 TI - Limitations of the Framingham risk score are now much clearer. PMID- 19124039 TI - Expression and localization of rabbit B-cell activating factor (BAFF) and its specific receptor BR3 in cells and tissues of the rabbit immune system. AB - Rabbits are widely used for vaccine development, and investigations of human infectious and autoimmune diseases such as Systemic Lupus Erythematosus (SLE). For these applications, we cloned, sequenced and expressed rabbit B-cell Activating Factor (BAFF), and localized BAFF in cells and tissues of the rabbit immune system. The rabbit homolog of the human BAFF binding site (miniBR3 peptide) within the BAFF-specific receptor BR3 was synthesized. This 26-residue core domain binds to recombinant rabbit BAFF protein. Flow cytometric analyses using purified recombinant rabbit BAFF combined with real-time PCR findings revealed that BAFF detected on peripheral blood B-cells from normal rabbits is probably complexed to BAFF receptors rather than produced by the B-cells. BAFF was detected in developing appendix of young rabbits by immunohistochemical staining suggesting that BAFF plays a role during the period following birth when rabbit B-cell development and pre-immune antibody repertoire diversification and selection is occurring. PMID- 19124040 TI - In vitro cultivation and cryopreservation of duck embryonic hepatocytes. AB - Hepatitis B-virucidal testing of biocides in quantitative suspension tests using duck hepatitis B virus (DHBV) requires primary duck embryonic hepatocytes for viral propagation. To improve the test system and availability of these cells, commercial culture plates with different growth surfaces were tested for cell cultivation and different approaches for cryopreservation of hepatocyte suspension were examined. After 12 days of culture, the largest amounts of hepatocytes were grown in CellBIND and TTP plates and CellBIND surface showed the lowest tendency of monolayer detachment nearly comparable with collagen 1-coated CELLCOAT plates. For cryopreservation of hepatocyte suspension, the use of growth medium supplemented with fetal calf serum (FCS) and dimethyl sulfoxide (ME(2)SO), FCS supplemented with ME(2)SO or cryosafe-1 as cryoprotective agents provided the highest rates of surviving cells after thawing. The freezing-thawing process did not significantly reduce the susceptibility of hepatocytes to infection with DHBV. In conclusion, plates without collagen 1 such as CellBIND are recommended for cultivation of primary duck embryonic hepatocytes in infectivity experiments of DHBV for virucidal testing of biocides. The use of cryopreserved hepatocytes is possible when freshly isolated cells from the liver of duck embryos are not available. PMID- 19124041 TI - Laboratory safe detection of nucleocapsid protein of Rift Valley fever virus in human and animal specimens by a sandwich ELISA. AB - A safe laboratory procedure, based on a sandwich ELISA (sAg-ELISA), was developed and evaluated for the detection of nucleocapsid protein (NP) of Rift Valley fever virus (RVFV) in specimens inactivated at 56 degrees C for 1h in the presence of 0.5% Tween-20 (v/v) before testing. Polyclonal capture and detection immune sera were generated respectively in sheep and rabbits immunized with recombinant NP antigen. The assay was highly repeatable and specific; it detected strains of RVFV from the entire distributional range of the disease, isolated over a period of 53 years; no cross-reactivity with genetically related African phleboviruses or other members of the family Bunyaviridae was observed. In specimens spiked with RVFV, including human and animal sera, homogenates of liver and spleen tissues of domestic ruminants, and Anopheles mosquito homogenates, the sAg-ELISA detection limit ranged from log(10)10(2.2) to 10(3.2) TCID(50)/reaction volume. The ELISA detected NP antigen in spiked bovine and sheep liver homogenates up to at least 8 days of incubation at 37 degrees C whereas infectious virus could not be detected at 48h incubation in these adverse conditions. Compared to virus isolation from sera from RVF patients and sheep infected experimentally, the ELISA had 67.7% and 70% sensitivity, and 97.97% and 100% specificity, respectively. The assay was 100% accurate when testing tissues of various organs from mice infected experimentally and buffalo foetuses infected naturally. The assay was able to detect NP antigen in infective culture supernatants 16-24h before cytopathic effects were observed microscopically and as early as 8h after inoculation with 10(5.8) TCID(50)/ml of RVFV. This ability renders the assay for rapid identification of the virus when its primary isolation is attempted in vitro. As a highly specific, safe and simple assay format, the sAg-ELISA represents a valuable diagnostic tool for use in less equipped laboratories in Africa, and for routine differential diagnosis of viral hemorrhagic fevers. PMID- 19124042 TI - Rapid detection and quantitation of poliovirus and rhinovirus sequences in viral stocks and infected cells. AB - Laboratories working with closely related viruses need simple and cost-effective ways to rapidly validate viral stocks, detect contamination and measure the abundance of viral RNA species. Using RT-PCR and specific primers an approach for the specific detection of rhinovirus type 14 (RV14) or poliovirus type 1 (PV1) is presented. It is demonstrated that viral sequences can be amplified directly from viral stocks or from infected cells. In addition, the utility of this protocol for the detection of low levels of contaminating PV1 in RV14 stocks is shown. Further, using quantitative real-time PCR It is shown that this approach can be used for the quantitative analysis of viral RNA and replication kinetics in infected cells. This method should be useful for laboratories working with PV and RV14 and could be adapted easily for use by laboratories working with other rhinovirus and enterovirus serotypes. PMID- 19124044 TI - Walking pattern analysis after unilateral 6-OHDA lesion and transplantation of foetal dopaminergic progenitor cells in rats. AB - Functional sensorimotor recovery after transplantation of mesencephalic dopaminergic (DAergic) neurons has been well documented in the rat 6 hydroxydopamine (6-OHDA) model of Parkinson's disease. However, the functional restoration of more specific gait-related patterns such as skilled walking, balance, and individual limb movements have been insufficiently studied. The purpose of this study was to investigate the behavioural effects of intrastriatal DA grafts on different aspects of normal and skilled walking in rats following unilateral 6-OHDA lesions of the medial forebrain bundle. Rats were subjected to drug-induced rotation, detailed footprint analysis, and assessment of skilled walking in the ladder rung walking test prior and after the transplantation of E14 ventral mesencephalon-derived progenitor cells. Good DAergic graft survival, as revealed by immunohistochemistry, was accompanied by a compensation of drug induced rotational asymmetries. Interestingly, the analysis of walking patterns displayed a heterogeneous graft-induced response in skilled and non-skilled limb use. Grafted animals made fewer errors with their contralateral limbs in skilled walking than the sham-transplanted rats, and they improved their ipsi- and contralateral limb rotation. However, the parameter distance between feet showed a delayed recovery, and the stride length was not affected by the DA grafts at all. These findings indicate that ectopic intrastriatal transplantation of E14 ventral mesencephalon-derived cells promotes recovery of gait balance and stability, but does not ameliorate the shuffling gait pattern associated with 6 OHDA lesions. A full restoration of locomotor gait pattern might require a more complete and organotypic reconstruction of the mesotelencephalic DAergic pathway. PMID- 19124043 TI - Estradiol induces region-specific inhibition of ZENK but does not affect the behavioral preference for tutored song in adult female zebra finches. AB - Female zebra finches display a preference for songs of males raised with tutors compared to those from males without tutors. To determine how this behavioral preference may be mediated by auditory perception sites, the social behavior network, and the dopamine reward system, and whether responses of these regions are affected by estradiol, females were treated with hormone or blank implants. An auditory choice test was conducted followed by exposure to tutored or untutored song or silence to examine induction of the immediate early gene, ZENK. Birds spent significantly more time near tutored than untutored song, regardless of estrogen treatment, and estradiol significantly decreased the density of ZENK immunoreactive neurons within the ventromedial hypothalamus. These results suggest that selective neural and behavioral responses can be induced by both high quality vocalizations and estradiol, although they are not necessarily correlated. PMID- 19124045 TI - The phytoestrogen ferutinin affects female sexual behavior modulating ERalpha expression in the hypothalamus. AB - This study was designed to assess the effect of the phytoestrogenic compound ferutinin, chronically administered in ovariectomized progesterone primed rats, alone or in combination with estradiol benzoate. After 2, 3 and 4 weeks of treatments, female rats were tested for receptive (lordosis) and proceptive behaviors (hops, darts and ear wigglings). Ferutinin given alone markedly increased the intensity of the lordotic response in ovariectomized rats but failed to significantly affect proceptivity. On the other hand estradiol benzoate significantly increased both receptive and proceptive behaviors. When administered in combination with estradiol, ferutinin reduced the increase in receptivity and proceptivity due to estrogen effects, acting as an antiestrogen. At the end of the behavioral experiments, animals were sacrificed and Western blot analysis of estrogen receptor alpha (ERalpha) levels was performed in the dissected hypothalami. Ferutinin increased ERalpha expression when administered alone, as estradiol did, but decreased the response to estradiol when administered in combination. These results suggest that ferutinin displays estrogenic or antiestrogenic activity through ERalpha in the hypothalamus, depending on the absence or the presence of estrogen priming. PMID- 19124046 TI - Molecular basis of multiple sulfatase deficiency, mucolipidosis II/III and Niemann-Pick C1 disease - Lysosomal storage disorders caused by defects of non lysosomal proteins. AB - Multiple sulfatase deficiency (MSD), mucolipidosis (ML) II/III and Niemann-Pick type C1 (NPC1) disease are rare but fatal lysosomal storage disorders caused by the genetic defect of non-lysosomal proteins. The NPC1 protein mainly localizes to late endosomes and is essential for cholesterol redistribution from endocytosed LDL to cellular membranes. NPC1 deficiency leads to lysosomal accumulation of a broad range of lipids. The precise functional mechanism of this membrane protein, however, remains puzzling. ML II, also termed I cell disease, and the less severe ML III result from deficiencies of the Golgi enzyme N acetylglucosamine 1-phosphotransferase leading to a global defect of lysosome biogenesis. In patient cells, newly synthesized lysosomal proteins are not equipped with the critical lysosomal trafficking marker mannose 6-phosphate, thus escaping from lysosomal sorting at the trans Golgi network. MSD affects the entire sulfatase family, at least seven members of which are lysosomal enzymes that are specifically involved in the degradation of sulfated glycosaminoglycans, sulfolipids or other sulfated molecules. The combined deficiencies of all sulfatases result from a defective post-translational modification by the ER localized formylglycine-generating enzyme (FGE), which oxidizes a specific cysteine residue to formylglycine, the catalytic residue enabling a unique mechanism of sulfate ester hydrolysis. This review gives an update on the molecular bases of these enigmatic diseases, which have been challenging researchers since many decades and so far led to a number of surprising findings that give deeper insight into both the cell biology and the pathobiochemistry underlying these complex disorders. In case of MSD, considerable progress has been made in recent years towards an understanding of disease-causing FGE mutations. First approaches to link molecular parameters with clinical manifestation have been described and even therapeutical options have been addressed. Further, the discovery of FGE as an essential sulfatase activating enzyme has considerable impact on enzyme replacement or gene therapy of lysosomal storage disorders caused by single sulfatase deficiencies. PMID- 19124047 TI - The serine hydroxymethyltransferase gene glyA in Corynebacterium glutamicum is controlled by GlyR. AB - Serine hydroxymethyltransferase (SHMT) occupies a central position in one-carbon metabolism, and we here study its regulation in Corynebacterium glutamicum. Enzyme quantifications revealed an about 3-fold increase of SHMT activity during exponential growth with a further increase at the onset of the stationary phase. The SHMT encoding glyA gene was shown to be transcribed as a monocistronic mRNA, and its transcriptional start site was determined. Using DNA affinity chromatography the regulator GlyR (Cg0527) was identified and its chromosomal deletion shown to abolish the increase in SHMT activity in the stationary phase. The involvement of GlyR in glyA control was further confirmed by a transcriptional fusion of the glyA promoter with cat and chloramphenicol acetyltransferase quantifications. GlyR was isolated and mutational studies together with electrophoretic mobility shift assays showed that it binds to the imperfect palindromic motif CACT-N(2)-AATG in the -119 to -96 upstream region of the glyA promoter. These and further data illustrate that the essential SHMT has highest activity in the stationary phase and that GlyR acts as an activator of glyA transcription in this growth phase. PMID- 19124048 TI - Genetic connection of two contiguous bacterial artificial chromosomes using homologous recombination in Bacillus subtilis genome vector. AB - A Bacillus subtilis genome (BGM) vector system using homologous recombination was applied to connect two contiguous BAC clones covering the entire 355-kb transcription unit of the mouse jumonji genomic region. Results from the convenient genomic manipulation indicated that the BGM system facilitates the connection of DNAs from a BAC library without exchange and deletion of original sequence, which can expand large-sized DNA construction beyond BAC-building in Escherichia coli. PMID- 19124049 TI - Deoxynojirimycin enhanced the transglycosylation activity of a glycosidase from the China white jade snail. AB - A beta-D-glycosidase (G I) from the China white jade snail showed non-Michaelis Menten mode in catalyzing the reaction using pNPGlu and pNPFuc as the substrate and monitoring the released pNP. We determined quantitatively both the transglycosidic and hydrolytic products of pNPGlu and pNPFuc solvolysis for the detailed kinetic analysis on G I-catalyzed hydrolysis and transglycosylation reaction. The inhibition kinetic studies using deoxynojirimycin (DNJ) and butanol as inhibitors were preceded. DNJ only inhibited competitively the hydrolysis of cellobiose and pNPGlu while "activated" the transglycosylation of pNPGlu and pNPFuc. This was evident from the increased V(max)tr value with no change of the apparent K(m)tr. In contrast, butanol exhibited a competitive inhibition to the transglycosylation reaction and non-competitive inhibition to the hydrolysis. The results indicated that the non-Michaelis-Menten kinetic behavior was caused by the co-occurrence of substrate transglycosylation reaction. This study provided a simple method to increase the transglycosylation yield by using DNJ to inhibit hydrolysis. PMID- 19124050 TI - Establishment of transgenic tobacco hairy roots expressing basic peroxidases and its application for phenol removal. AB - Transgenic hairy root (HR) systems constitute an interesting alternative to improve the efficiency of phytoremediation process. Since peroxidases (Px) have been associated with phenolic compounds removal, in the present work, transgenic tobacco HR, which expressed basic Px genes from tomato (tpx1 and tpx2), were established and assayed for phenol removal. Tobacco HR clones were obtained, including those transgenic for TPX1 or TPX2, those double transgenic (DT) for both Px and the corresponding controls. Based on growth index, the presence of rol C sequence, tpx1 and/or tpx2 genes and the coded proteins, as well as Px activity determinations, we selected 10 tobacco HR clones for phenol removal assays. The removal efficiencies were high for all the HR, although, some transgenic HR showed significantly higher removal efficiencies compared with controls. The results demonstrate that TPX1 is involved in phenol removal not only when it was overexpressed in tomato, but also when it was expressed in other plant, such as tobacco. The higher efficiency of TPX2 transgenic HR showed that this Px also participates in the process. The contribution of other mechanisms (adsorption, H2O2 independent enzymatic processes) could be considered depreciable, which establishes the great implication of Px in phenol removal. PMID- 19124051 TI - Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC). AB - Liposomal formulation of siRNA is an attractive approach for improving its delivery in vivo, shielding the RNA from nucleases and promoting tumor targeting. Here, the production of very small batch sizes of siRNA-liposomes by using the "dual asymmetric centrifugation (DAC)" technique was investigated. This new technique combines rapid and sterile liposome preparation with very high entrapping efficiencies. DAC is here presented in conjunction with a non destructive microscale analysis based on double fluorescence labeling, which enables monitoring of siRNA integrity during the liposomal preparation. Integrity is reflected in spatial proximity of the dyes, which results in measurable fluorescence resonance energy transfer (FRET). The combination of DAC and the sensitive FRET analysis allows the handling of batch sizes down to 20 mg of conventional liposomes (CL) and sterically stabilized liposomes (SL). These were prepared in common 2 ml reaction tubes and loaded with calcein or labeled siRNA. Liposome sizes were 79+/-16 nm for CL and 109+/-9 nm for SL loaded with siRNA. Trapping efficiencies ranged from 43 to 81%, depending on batch size, enclosed compound, and liposome composition. FRET monitoring showed that the siRNA remained intact throughout DAC and that liposomal formulations protected the siRNA from nucleases. siRNA-liposomes remained stable for at least 3 months. PMID- 19124052 TI - The role of human ABC transporter ABCG2 (BCRP) in pharmacotherapy. PMID- 19124053 TI - Purification and structural analyses of ABCG2. AB - ABCG2 is best known as a multidrug transporter capable of conferring resistance to cancer cells. However, the protein is also inherently expressed in numerous barrier tissues and intriguingly within hematopoietic stem cells. Unlike its partners ABCB1 and ABCC1, there is considerably less information available on the molecular mechanism of ABCG2. The transporter has a distinct topology and is presumed to function as a homodimer. However, a number of biochemical studies have presented data to suggest that the protein adopts higher order oligomers. This review focuses on this controversial issue with particular reference to findings from low resolution structural data. In addition, a number of molecular models of ABCG2 based on high resolution structures of bacterial ABC transporters have recently become available and are critically assessed. ABCG2 is a structurally distinct member of the triumvirate of human multidrug transporters and continues to evade description of a unifying molecular mechanism. PMID- 19124054 TI - Controlled coupling of peptides at their C-termini. AB - Fusion of two proteins has become an important tool in biotechnology. Whereas biotechnological methods easily can produce C-terminal to N-terminal fused compounds, methods to couple two proteins to each of their C-termini are not easily accessible. Herein, peptides are used as models for larger proteins. A method is described exploiting the possibility to attach different reactive handles to their C-termini using a reaction catalyzed by the enzyme carboxypeptidase Y (CPY). It is possible to attach pairs of reaction handles which can react with each other to each of the peptides to be coupled. In a second step, the two modified peptides can be linked together by a chemical reaction, such as an oxime-forming reaction or a copper(I) catalyzed [2+3] cycloaddition reaction of an azide with an alkyne. PMID- 19124055 TI - Role and action in the pituitary corticotroph of corticotropin-releasing factor (CRF) in the hypothalamus. AB - Corticotropin-releasing factor (CRF), produced in the hypothalamic paraventricular nucleus (PVN) in response to stress, stimulates the synthesis and secretion of adrenocorticotropin (ACTH) via CRF receptor type 1 (CRF(1) receptor) in the anterior pituitary (AP) of mammals. CRF is critical for the circadian rhythmicity of the hypothalamic-pituitary-adrenal axis and the augmented release of ACTH from the pituitary in response to the stress. A higher molecular weight form of immunoreactive beta-endorphin, putative proopiomelanocortin (POMC), is increased in CRF-knockout mice (CRF KO), suggesting the important role of CRF in the processing of POMC. In fact, CRF is able to modulate the processing of POMC through changes in prohormone convertase (PC)-1 expression levels. Multiple forms of ACTH-related peptides containing unprocessed ones are present in some cases of ACTH-producing tumors, presumably without action of PC-1 under the control of CRF. Following CRF-activated stimulation of the receptor signaling, CRF(1) receptor is down-regulated and desensitized. In fact, CRF facilitates the degradation of CRF(1) receptor mRNA via the protein kinase A pathway. Prolonged agonist activation of CRF(1) receptor leads to a loss of responsiveness, or desensitization of the receptor. G protein-coupled receptor kinase 2 is involved in desensitization of CRF(1) receptor by CRF in the corticotroph. PMID- 19124056 TI - Clonal diversity of cytotoxic T lymphocytes that recognize autologous oral squamous cell carcinoma. AB - Cytotoxic T lymphocytes (CTLs) play an essential role in immunologic responses for tumor rejection. In the past decade, various melanoma tumor-associated antigens (TAAs) have been identified, and several clinical trials of vaccination immunotherapy and adoptive immunotherapy using such antigens with or without adjuvants have had fascinating results. However, this has not been the case with oral squamous cell carcinoma (OSCC) because of the difficulty of establishing oral cancer cell lines and CTLs against autologous oral cancer cells. Therefore, few oral cancer antigens have been identified with such CTLs. We herein present the successful establishment of an oral squamous cell carcinoma cell line, POT-1, and an HLA-A24-restricted CTL line (TcPOT-1) from a patient's autologous peripheral blood lymphocytes. TcPOT-1 recognized autologous POT-1 cells in an HLA A24-restricted manner, and also allogeneic HLA-A24 (+) OSCC cell lines OSC-70 and HSC-2. We also succeeded in isolating two distinct CTL clones from TcPOT-1, HLA A24-restricted CTL clone 4F11 and HLA-A33-restricted clone 4A11. Both of these clones recognized autologous POT-1 but not allogeneic OSSC cell lines. These data imply that the TcPOT-1 CTL line may include several CTL subpopulations with distinct antigen specificities, such as an HLA-A24-restricted POT-1-specific clone, HLA-A33-restricted POT-1-specific clone, and HLA-A24-restricted allogeneic OSCC-recognizing clone. Therefore, precise analysis of TcPOT-1-recognizing antigens may provide us with important information on as-yet-unknown tumor rejection antigens in OSCC. PMID- 19124057 TI - Safety and immunogenicity of a hexavalent diphtheria-tetanus-acellular pertussis inactivated poliovirus-Haemophilus influenzae b conjugate-hepatitis B vaccine at 2, 3, 4, and 12-14 months of age. AB - Combination vaccines improve parental and provider satisfaction and schedule compliance by decreasing the number of injections. In a Phase 2, randomized, double-blind, multicenter study, we compared four formulations of a liquid, hexavalent diphtheria-tetanus-acellular pertussis-inactivated poliovirus Haemophilus influenzae b conjugate-hepatitis B virus (DTaP-IPV-Hib-HBV) vaccine in 708 infants immunized at 2, 3, 4, and 12-14 months of age. The formulations contained identical DTaP and IPV components, differing in the contents of Hib polyribosylribitol phosphate (PRP) conjugate component (tetanus-toxoid [PRP-T, 12microg] or Neisseria meningitidis outer-membrane-protein-complex [PRP-OMPC, 3microg or 6microg]), and in hepatitis B surface antigen (HBsAg, 10microg or 15microg). A minimum acceptable postdose 3 antibody response rate was defined by the lower limit of the 95% confidence interval exceeding a prespecified target. Rates of adverse events (AEs) were similar among groups, with a trend for increased solicited injection-site reactions (pain, redness, swelling) with increasing PRP-OMPC and HBsAg concentration. Serious AEs reported by eight subjects were not considered to be vaccine related. All PRP-OMPC formulations met prespecified acceptability criteria for postdose 3 immunogenicity for all antigens: PRP, HBsAg, pertussis, diphtheria, tetanus and polio. Apart from the Hib response, the postdose 3 responses obtained with the PRP-T formulation met the acceptability criterion for each antigen. Postdose 4 responses were acceptable for all antigens in all formulations. All vaccine formulations were well tolerated. The three PRP-OMPC formulations met prespecified immunogenicity criteria, and the one with the lowest PRP-OMPC concentration was selected for further optimization of immunogenicity. PMID- 19124059 TI - Antioxidant effects of flavonoid from Croatian Cystus incanus L. rich bee pollen. AB - Oxidant/antioxidant status, estrogenic/anti-estrogenic activity and gene expression profile were studied in mice fed with Cystus incanus L. (Cistaceae) reach bee pollen from location in Central Croatia's Dalmatia coast and offshore islands. Seven phenolic compounds (out of 13 tested) in bee pollen sample were detected by high performance liquid chromatography (HPLC) analysis. Phenolics detected in C. incanus L. bee pollen belong to flavonol (pinocembrin), flavanols (quercetin, kaempferol, galangin, and isorhamnetin), flavones (chrysin) and phenylpropanoids (caffeic acid). Bee pollen as a food supplement (100mg/kgbw mixed with commercial food pellets) compared to control (commercial food pellets) modulated antioxidant enzymes (AOE) in the mice liver, brain and lysate of erythrocytes and reduced hepatic lipid peroxidation (LPO). Bee pollen induced 25% of anti-estrogenic properties while no estrogenic activity was found. Differential gene expression profile analyses after bee pollen enriched diet identify underexpressed gene Hspa9a, Tnfsf6 (liver) and down-regulated gene expression of Casp 1 and Cc121c (brain) which are important in the apoptosis pathway and chemotaxis. These results indicate that used bee pollen possess a noticable source of compounds with health protective potential and antioxidant activity. PMID- 19124060 TI - Viewing brain processes as Critical State Transitions across levels of organization: Neural events in Cognition and Consciousness, and general principles. AB - In this theoretical and speculative essay, I propose that insights into certain aspects of neural system functions can be gained from viewing brain function in terms of the branch of Statistical Mechanics currently referred to as "Modern Critical Theory" [Stanley, H.E., 1987. Introduction to Phase Transitions and Critical Phenomena. Oxford University Press; Marro, J., Dickman, R., 1999. Nonequilibrium Phase Transitions in Lattice Models. Cambridge University Press, Cambridge, UK]. The application of this framework is here explored in two stages: in the first place, its principles are applied to state transitions in global brain dynamics, with benchmarks of Cognitive Neuroscience providing the relevant empirical reference points. The second stage generalizes to suggest in more detail how the same principles could also apply to the relation between other levels of the structural-functional hierarchy of the nervous system and between neural assemblies. In this view, state transitions resulting from the processing at one level are the input to the next, in the image of a 'bucket brigade', with the content of each bucket being passed on along the chain, after having undergone a state transition. The unique features of a process of this kind will be discussed and illustrated. PMID- 19124061 TI - New (but old) molecules regulating synapse integrity and plasticity: Cbln1 and the delta2 glutamate receptor. AB - The delta2 glutamate receptor (GluRdelta2) is predominantly expressed in cerebellar Purkinje cells and plays crucial roles in cerebellar functions: GluRdelta2-null mice display ataxia and impaired motor learning. Interestingly, the contact state of synapses between parallel fibers (PFs) and Purkinje cells is specifically and severely affected, and the number of normal PF synapses is markedly reduced in GluRdelta2-null Purkinje cells. Furthermore, long-term depression at PF-Purkinje cell synapses is abrogated. Cbln1, a member of the C1q/tumor necrosis factor (TNF) superfamily, is predominantly expressed and released from cerebellar granule cells. Unexpectedly, the behavioral, physiological and anatomical phenotypes of cbln1-null mice precisely mimic those of GluRdelta2-null mice. Thus, we propose that Cbln1, which is released from granule cells, and GluRdelta2, which is predominantly expressed in Purkinje cells, are involved in a common signaling pathway crucial for synapse formation/maintenance and plasticity in the cerebellum. Since molecules related to Cbln1 are expressed in various brain regions other than the cerebellum, other C1q/TNF superfamily proteins may also regulate various aspects of synapses in the CNS. Therefore, an understanding of the signaling mechanisms underlying Cbln1 and GluRdelta2 in the cerebellum will provide new insights into the roles of C1q/TNF superfamily proteins as new cytokines that regulate normal and abnormal brain functions. PMID- 19124062 TI - Analyzing variable behavioral contingencies: are certain complex skills homologous with locomotion? AB - This paper considers behavioral contingencies that change as a function of time, of the individual's own behavior (as in locomotion and reading), of the behavior of other parties or of interactions with them. A detailed analysis of locomotion and of reading out loud shows that the behavioral contingencies for these are virtually the same. The terrain being traversed and the locomotion behavior involved are shown to be analogous to a segment of text being read and the articulation of the words. In both cases, successive upcoming segments are perceived and processed, and during the processing phases, motor behavior is formulated. In both, the smooth concatenation of the motor phases for successive segments requires buffering. Both involve corrective or digressive actions in response to obstacles or unanticipated stimuli encountered. Both involve looking ahead at the upcoming segment and processing it while the motor phase of the prior segment is still in progress. For both, the size, entropy, familiarity, and other attributes of the upcoming segment are parameters of the performance. It is suggested that locomotion has similar parallels with certain other complex skills, such as listening, copying, receiving Morse code, simultaneous interpreting, and certain types of performance, and may therefore be their phylogenetic prototype and biological homologue. PMID- 19124063 TI - Relationships between replication timing and GC content of cancer-related genes on human chromosomes 11q and 21q. AB - The human genome is composed of large-scale compartmentalized structures, including long G+C% (GC%) mosaic structures and replication-timing zones, which are related to chromosome band zones. Previously, we measured replication timing along the entire lengths of human chromosomes 11q and 21q at the sequence level, and it was suggested that the transition regions of replication timing from early to late S-phase coincided with "unstable" regions of the genome associated with increased DNA damage. In the present study, we measured replication timing of 15 known oncogenes and tumor suppressor genes on human chromosomes 11q and 21q using two human cell lines (THP-1 and Jurkat). We found unusual relationships between replication timing and the GC content of the genomic regions in which these cancer-related genes were located. Many of these genes showed similar replication timing between the two cell lines, and the majority replicated intermediately between early and late in both cell lines. On the other hand, more than half of these genes were located at very GC-rich (50-55 GC%) regions. In addition, we analyzed the exact relationships between early/late-switch regions of replication timing where cancer-related genes were located, and GC% transitions in and around five R/G-chromosomal band boundaries (each ca. 4 Mb) by using newly designed PCR primer sets. We found that the majority of cancer-related genes including oncogenes were located in GC-rich isochores close to GC% transitions within early/late-switch regions of replication timing, many of which replicated intermediately between the early and late S-phase. Unusual relationships between replication timing and GC content of the genomic regions in which cancer-related genes were located may be related to the molecular mechanisms of genomic instability associated with increased DNA damage. PMID- 19124064 TI - The Bcl-w promoter is activated by beta-catenin/TCF4 in human colorectal carcinoma cells. AB - The antiapoptotic BCL-2 family protein BCL-W is often overexpressed in colorectal carcinoma (CRC) where it correlates with advanced stage and expression of p53. In this work we have analysed the Bcl-w promoter to identify potential regulators of BCL-W expression in CRC cells. The Bcl-w promoter was highly active in cell lines derived from CRC as well as other cancer types. Although expression of p53 and BCL-W correlate in CRC, overexpression of wild type or mutant p53 did not significantly alter Bcl-w promoter activity, and deletion of endogenous p53 did not alter the expression of Bcl-w RNA in HCT116 cells. Promoter deletion analysis lead to the identification of a potential binding site for TCF/LEF factors, obligate binding partners for beta-catenin, a downstream target of the WNT signalling pathway. TCF4 and beta-catenin interacted with the Bcl-w promoter in intact HCT116 cells and mutation of this site significantly decreased promoter activity. The activity of the Bcl-w promoter was increased or decreased, respectively, by overexpression of beta-catenin or dominant negative TCF4. beta catenin is activated in the majority of CRC and these results suggest that BCL-W may function as a downstream effector of inappropriate WNT/beta-catenin signalling. PMID- 19124065 TI - Enhancement of oral absorption of curcumin by self-microemulsifying drug delivery systems. AB - Curcumin is a poorly water-soluble drug and its oral bioavailability is very low. A new self-microemulsifying drug delivery system (SMEDDS) has been successfully developed to improve the solubility and oral absorption of curcumin. Suitable compositions of SMEDDS formulation were screened via solubility studies of curcumin and compatibility tests. The formulation of curcumin-loaded SMEDDS was optimized by a simplex lattice experiment design. The optimal formulation of SMEDDS was comprised of 57.5% surfactant (emulsifier OP:Cremorphor EL = 1:1), 30.0% co-surfactant (PEG 400) and 12.5% oil (ethyl oleate). The solubility of curcumin (21 mg/g) significantly increased in SMEDDS. The average particle size of SMEDDS-containing curcumin was about 21 nm when diluted in water. No significant variations in particle size and curcumin content in SMEDDS were observed over a period of 3 months at 4 degrees C. The spherical shape of microemulsion droplet was observed under TEM. The dissolution study in vitro showed that more than 95% of curcumin in SMEDDS could be dissolved in pH 1.2 or pH 6.8 buffer solutions in 20 min, however, less than 2% for crude curcumin in 60 min.The in situ absorption property of curcumin-loaded SMEDDS was evaluated in intestines of rats. The results showed the absorption of curcumin in SMEDDS was via passive transfer by diffusion across the lipid membranes. The results of oral absorption experiment in mice showed that SMEDDS could significantly increase the oral absorption of curcumin compared with its suspension. Our study illustrated that the developed SMEDDS formulation held great potential as a possible alternative to traditional oral formulations of curcumin. PMID- 19124066 TI - Modeling a system of phosphated cross-linked high amylose for controlled drug release. Part 2: physical parameters, cross-linking degrees and drug delivery relationships. AB - High amylose cross-linked to different degrees with sodium trimetaphosphate by varying base strength (2% or 4%) and contact time (0.5-4h) was evaluated as non compacted systems for sodium diclophenac controlled release. The physical properties and the performance of these products for sodium diclophenac controlled release from non-compacted systems were related to the structures generated at each cross-linking degree. For samples at 2% until 2h the swelling ability, G' and eta* values increased with the cross-linking degree, because the longer polymer chains became progressively more entangled and linked. This increases water uptake and holding, favoring the swelling and resulting in systems with higher viscosities. Additionally, the increase of cross-linking degree should contribute for a more elastic structure. The shorter chains with more inter-linkages formed at higher cross-linking degrees (2%4h and 4%) make water caption and holding difficult, decreasing the swelling, viscosity and elasticity. For 2% samples, the longer drug release time exhibited for 2%4h sample indicates that the increase of swelling and viscosity contribute for a more sustained drug release, but the mesh size of the polymeric network seems to be determinant for the attachment of drug molecules. For the 4% samples, smaller meshes size should determine less sustained release of drug. PMID- 19124067 TI - Zn(2+), derived from cell preparation, partly attenuates Ca(2+)-dependent cell death induced by A23187, calcium ionophore, in rat thymocytes. AB - A23187, a calcium ionophore, is used to induce Ca(2+)-dependent cell death by increasing intracellular Ca(2+) concentration ([Ca(2+)](i)) under in vitro condition. Since this ionophore also increases membrane permeability of metal divalent cations such as Zn(2+) and Fe(2+) rather than Ca(2+), trace metal cations in cell suspension may affect Ca(2+)-dependent cell death induced by A23187. Therefore, the effects of chelators for divalent metal cations, EDTA and TPEN, on the A23187-induced cytotoxicity were cytometrically examined in rat thymocytes. The cytotoxicity of A23187 was attenuated by 1mM EDTA while it was augmented by 50 microM EDTA and 10 microM TPEN. These changes were statistically significant. The A23187-induced increase in Fluo-3 fluorescence intensity, a parameter for [Ca(2+)](i), was significantly reduced by 1mM EDTA while it was not the case for 50 microM EDTA and 10 microM TPEN. The intensity of FluoZin-3 fluorescence, a parameter for [Zn(2+)](i), increased by A23187 was respectively reduced by 50 microM EDTA and 10 microM TPEN. It is suggested that the attenuation of A23187-induced cytotoxicity by 1mM EDTA is due to the chelation of extracellular Ca(2+) and Zn(2+) while the augmentation by 50 microM ETDA or 10 microM TPEN is due to the chelation of extracellular Zn(2+). The Tyrode's solution without thymocytes contained 32.4 nM of zinc while it was 216.9 nM in the cell suspension. In conclusion, trace Zn(2+), derived from cell preparation, partly attenuates the Ca(2+)-dependent cell death induced by A23187. PMID- 19124068 TI - The use of established skeletal muscle cell lines to assess potential toxicity from embedded metal fragments. AB - The use of novel materials on the modern battlefield, both in military munitions as well as in Improvised Explosive Devices, opens the possibility of wounds with embedded fragments whose health effects and toxicity characteristics have not been fully investigated, if at all. The costly and time-consuming nature of standard two-year lifespan studies prohibits the testing of many materials. In this report, we describe an in vitro system for rapidly assessing potential toxicity of metals and metal mixtures. Using rat L6 and mouse C2C12 skeletal muscle cells and tests for cellular viability, we have shown that two militarily relevant tungsten alloy mixtures (W/Ni/Co and W/Ni/Fe) significantly decreased the metabolic viability of rat L6 cells, whereas the viability of mouse C2C12 cells was not affected by W/Ni/Co and only slightly affected by W/Ni/Fe. In addition, viability assessed through lysosomal uptake of neutral red dye was not affected by either mixture in either cell line indicating that the mitochondria may be the target organelle of these unique metal mixtures. Development of this in vitro screening system may provide a procedure by which the potential toxicities of embedded metal fragments can be rapidly assessed. PMID- 19124069 TI - Genotoxicity of plumbagin and its effects on catechol and NQNO-induced DNA damage in mouse lymphoma cells. AB - Plumbagin, a naphtoquinone present in the roots of Plumbago zeylanica, has been reported to have many beneficial effects such as antibacterial, antifungal, anticancer, antimutagenic and antioxidant effects, but this compound has also been reported to have many side effects. Given the wide use of P. zeylanica in traditional medicine and the various potential therapeutic uses of plumbagin, the present study was carried out to further elucidate the potential genotoxicity and antigenotoxicity of plumbagin in mouse lymphoma L5178Y cells, using the comet assay. Without affecting the cell viability, plumbagin itself was found to induce significant DNA damage at concentrations as low as 0.25 ng/ml. When the cells were exposed to non-DNA damaging concentrations of plumbagin, together with NQNO (known to interact with DNA in many different ways) or catechol (known to induce oxidative DNA damage), plumbagin was found to significantly reduce the catechol induced DNA damage, but to be without protective effect against the NQNO-induced damage. The fact that non-DNA damaging concentrations of plumbagin diminished the DNA damage induced by catechol, provides further support for the idea that plumbagin may act as an antioxidative agent at low concentrations. PMID- 19124070 TI - Naringenin-induced apoptosis is attenuated by Bcl-2 but restored by the small molecule Bcl-2 inhibitor, HA 14-1, in human leukemia U937 cells. AB - Naringenin, a naturally occurring citrus flavonone, has shown cytotoxicity in various human cancer cell lines as well as inhibitory effects on tumor growth and there is increasing interest in its therapeutic applications. In this study, the effect of ectopic Bcl-2 expression on naringenin-induced apoptosis was investigated. We found that Bcl-2 overexpression markedly protected human leukemia U937 cells from time- and dose-dependent induction of apoptosis by naringenin, as did caspase-3 and caspase-9 inhibitors. Additionally, Bcl-2 overexpression attenuated naringenin-induced Bax translocation and cytosolic release of cytochrome c. Our results also indicated that co-administration of HA14-1 and naringenin increased apoptosis in Bcl-2 overexpressing U937 cells by restoring mitochondrial dysfunction and activation of caspase-9 and caspase-3, as well as by cleavage of poly (ADP-ribose) polymerase. Taken together, these observations indicate that Bcl-2 confers apoptosis resistance to naringenin by inhibiting a mitochondrial amplification step in U937 cells. PMID- 19124071 TI - Rapid and sensitive detection of white spot syndrome virus by loop-mediated isothermal amplification combined with a lateral flow dipstick. AB - Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acids under isothermal conditions using a set of four specifically designed primers that recognize six distinct target sequences. It can be combined with a chromatographic lateral flow dipstick (LFD) for highly specific, rapid and simple visual detection of WSSV-specific amplicons. Using this protocol, a 30-min amplification followed by 5 min hybridization with an FITC-labeled DNA probe and 5 min LFD resulted in visualization of DNA amplicons trapped at the LFD test line. Thus, 10 min for rapid DNA extraction followed by LAMP combined with LFD detection resulted in a total assay time of approximately 50 min. Detection sensitivity was comparable to other commonly-used methods for nested PCR detection of WSSV but had the additional advantages of reduced assay time, confirmation of amplicon identity by hybridization and elimination of electrophoresis with carcinogenic ethidium bromide. PMID- 19124072 TI - Localization of the transcription factor, sterol regulatory element binding protein-2 (SREBP-2) in the normal rat brain and changes after kainate-induced excitotoxic injury. AB - The transcription factor sterol regulatory element binding protein-2 (SREBP-2) has a key role in regulating cholesterol biosynthesis. In view of ongoing cholesterol biosynthesis in the brain, the present study was carried out to examine the distribution of the SREBP-2 in the normal rat brain, and possible changes after kainate-induced excitotoxicity. Western immunoblot analysis of SREBP-2 showed a dense band at 70kDa corresponding to the transcriptionally active form of SREBP-2 in homogenates from the rat hippocampus, cortex and striatum. SREBP-2 immunolabeled sections showed dense labeling of pyramidal neurons in field CA1 and CA3 of the hippocampus, moderately dense labeling of pyramidal and non-pyramidal neurons of the cerebral neocortex, and moderate labeling of putative medium spiny neurons in the caudate nucleus and putamen. Label was observed in the somatic cytoplasm, nucleus and apical dendrites of pyramidal neurons, and dendritic shafts and spines in the neuropil in the hippocampus. The expression of SREBP-2 was also elucidated after excitotoxic neuronal injury induced by intracerebroventricular injections of kainate. Real time RT-PCR analyses showed that mRNA level of SREBP-2 was significantly reduced at both 1 day and 2 weeks post-kainate injection. Immunohistochemical analyses showed significantly reduced SREBP-2 immunoreactivity in the kainate-induced lesions, in support of the real time RT-PCR results. Taken together, the above results show that there is high level of SREBP-2 expression in the normal hippocampus, and that neuronal injury results in a significant reduction of SREBP 2 expression in the damaged areas. PMID- 19124073 TI - Galanin-like peptide stimulates feeding and sexual behavior via dopaminergic fibers within the medial preoptic area of adult male rats. AB - Galanin-like peptide (GALP) is located in the arcuate nucleus (Arc) of the hypothalamus and is known to regulate both food intake and sexual behaviors in adult male rats. We have previously demonstrated that ICV GALP administration elicits a significant fos response within the medial preoptic area (mPOA). GALP is known to stimulate both food intake and male-typical sex behavior, presumably by direct actions within the mPOA. Recent data from our and other labs have led us to suspect that GALP effects on sex behaviors are due to activation of incertohypothalamic dopaminergic neurons that terminate within the mPOA. To test the hypothesis that GALP activates mPOA dopaminergic systems, we utilized an immunolesion technique to eliminate dopaminergic fiber input to the mPOA via a dopamine transporter-specific toxin (DATSAP, n=8) and compared to control injections (SAP, n=8). All animals were sexually experienced adult male Long Evans rats. DATSAP-treated male rats showed a significant (p<0.001) reduction in male sexual behaviors compared to SAP controls. We found that elimination of dopaminergic fibers within the mPOA significantly (p<0.001) eliminated all aspects of male sexual behavior under normal mating paradigms. Injections of GALP (5.0 nmol) significantly increased (p<0.01) male sex behavior and food intake in SAP control male rats but GALP did not stimulate the expression of these behaviors in DATSAP-treated rats. The orexigenic and anorexigenic effects of GALP were significantly (p<0.001) attenuated in DATSAP-treated male rats compared to SAP controls; however, ICV GALP was still able to significantly (p<0.05) reduce 24h body weight in both DATSAP and SAP rats. ICV GALP significantly (p<0.05) stimulated fos within the mPOA of SAP rats but not in DATSAP-treated male rats. These data suggest that GALP activates feeding and sexual behaviors in male rats by stimulating dopaminergic neurons that terminate within the mPOA. PMID- 19124076 TI - Injectability of biodegradable in situ forming microparticle systems (ISM). AB - In situ forming microparticle (ISM) systems offer a new encapsulation technique that provide a prolonged release of drug along with much greater ease of preparation and administration than conventional microparticles and surgically implanted systems. In this study, the forces required to inject biodegradable emulsions of the ISM systems via different sizes of syringe and needle into a newly developed chicken meat model, were determined using a compression mode of Texture Analyzer. The obtained injection forces were finally correlated with the injectability of the formulations into living rats. Results indicated that the flow of the ISM formulations through the needle could be described by the well known Poiseuille equation. The injection forces were directly proportional to the formulations' viscosity, injection speed, the second power of the syringe plunger radius and inversely proportional to the forth power of the needle radius. This test method could differentiate very well the required forces from various formulations and injection conditions as well as different injection sites (muscle, subcutaneous tissues or air). In comparison to the polymer solutions that form implants in situ, the ISM systems were more easily injectable with smaller needle size thus expected to be less painful and give better patient comfort/compliance. PMID- 19124075 TI - Support vector machine and pharmacophore-based prediction models of multidrug resistance protein 2 (MRP2) inhibitors. AB - Overexpression of multidrug-resistance protein 2 (MRP2) is one of the main causes that lead the curative effect reduction of many drugs, particularly anticancer drugs. Development of MRP2 inhibitors is the aim to overcome multidrug resistance due to MRP2. In this study, computational prediction models of MRP2 inhibitors have been developed by using support vector machine (SVM) and pharmacophore modeling method. For the SVM model, the overall prediction accuracy is 82.9% for the training set (257 compounds) and 77.1% for the independent test set (61 compounds). And 16 descriptors have been used in the SVM modeling; but from which it is difficult to get understanding about the action mechanism. The established pharmacophore model Hypo1 consists of two hydrogen bond acceptors and one hydrophobic feature. With the use of Hypo1, 78.1% of MRP2 inhibitors and 69.6% non-inhibitors can be predicted correctly. The overall prediction accuracy is 73.9%. Although the prediction accuracy of the pharmacophore model is lower than that of SVM model, it gives a clear picture of chemical features necessary for the MRP2 inhibitors. Taken together, the SVM model is capable of predicting MRP2 inhibitors with considerable good accuracy. But the gain of action mechanism related information needs the help of pharmacophore model. PMID- 19124074 TI - Genetic deficiency of NADPH oxidase does not diminish, but rather enhances, LPS induced acute inflammatory responses in vivo. AB - Reactive oxygen species (ROS) and oxidative stress are thought to play a central role in the etiology of cell dysfunction and tissue damage in sepsis. However, there is limited and controversial evidence from in vivo studies that ROS mediate cell signaling processes that elicit acute inflammatory responses during sepsis. Because NADPH oxidase is one of the main cellular sources of ROS, we investigated the role of this enzyme in lipopolysaccharide (LPS)-induced acute inflammation in vivo, utilizing mice deficient in the gp91(phox) or p47(phox) subunits of NADPH oxidase. Age-and body weight-matched C57BL/6J wild-type (WT) and gp91(phox-/-) and p47(phox-/-) mice were injected ip with 50 microg LPS or saline vehicle and sacrificed at various time points up to 24 h. We found that LPS-induced acute inflammatory responses in serum and tissues were not significantly diminished in gp91(phox-/-) and p47(phox-/-) mice compared to WT mice. Rather, genetic deficiency of NADPH oxidase was associated with enhanced gene expression of inflammatory mediators and increased neutrophil recruitment to lung and heart. Furthermore, no protection from LPS-induced septic death was observed in either knockout strain. Our findings suggest that NADPH oxidase-mediated ROS production and cellular redox signaling do not promote, but instead limit, LPS-induced acute inflammatory responses in vivo. PMID- 19124077 TI - Localization and quantification of biodegradable particles in an intestinal cell model: the influence of particle size. AB - The physicochemical parameters influencing particle crossing through the epithelial barrier have been largely studied and size has been shown as a crucial factor. Most part of this work has been done with polystyrene (PS) particles; however, few studies were conducted with well characterized biodegradable polymeric nanoparticles, more suitable for drug delivery. The aim of the present work was to study the influence of the size of well characterized biodegradable polymeric particles on cellular uptake by Caco-2 cells. Poly (d,l-lactide-co glycolide acid) (PLGA) particles loaded with a fluorescent dye, 3,3' dioctadecyloxacarbo-cyanine perchlorate (DiO), were prepared by the emulsion evaporation process. Five batches of particles with narrow size distribution (100, 300, 600, 1000, and 2000nm) were produced using selective centrifugation. Surface properties (zeta potential, hydrophobicity and residual surfactant rate) were similar among all batches. Preliminary study showed that after growth and differentiation, cell counting differed from plate to plate. Therefore, a quantitative method using fluorescence spectroscopy was developed to estimate the number of particles interacting with a single cell. The results were compared to the interaction obtained with polystyrene particles, commonly used as particle model. The interaction was clearly dependant on particle size and concentration. Particles in the range of 100nm presented a higher interaction when compared to larger particles. More than 6000 (d,l-lactide-co-glycolide acid) particles and 200,000 polystyrene particles were quantified per cell. Cellular localisation of particles by confocal microscopy showed the association of the poly (d,l-lactide co-glycolide acid) particles with the cells. Small particles were observed intracellularly, whereas particles larger than 300nm were associated with the apical membranes. Interestingly, some of the 100nm PLGA particles were localized in the nuclei of the cells. PMID- 19124078 TI - Innexin genes and gap junction proteins in the locust frontal ganglion. AB - Gap junctions (GJs) belong to one of the most conserved cellular structures in multicellular organisms. They probably serve similar functions in all Metazoa, providing one of the most common forms of intercellular communication. GJs are widely distributed in embryonic cells and tissues and have been attributed an important role in development, modulating cell growth and differentiation. These channels have been also implicated in mediating electrical synaptic signaling; Coupling through GJs is now accepted as a major pathway that supports network behavior and contributes to physiological rhythms. Here we focus on the physiology and molecular biology of GJs in a recently established model for the study of rhythm-generating networks and their role in behavior: the frontal ganglion (FG) of the desert locust, Schistocerca gregaria. Four novel genes of the invertebrate GJs (innexin) gene family were found to be expressed in the FG: Sg-inx1, Sg-inx2, Sg-inx3 and Sg-inx4. Immunohistochemistry revealed that some of the neurons in the FG express at least one innexin protein, INX1. We also established the presence of functional gap junction proteins in the FG and demonstrated functional electrical coupling between the neurons in the FG. This study forms the basis for further investigation of the role of GJs in network development and behavior. PMID- 19124079 TI - Is thrombin generation the new rapid, reliable and relevant pharmacological tool for the development of anticoagulant drugs? AB - The ex vivo testing emerges as an essential and critical step for the selection of the most promising prospective anticoagulant agents. The aim of the present study was to validate the thrombin generation assay as an ex vivo pharmacological screening test for measuring the anticoagulant behaviour and potency of molecules. The effects of six thrombin and/or factor Xa (FXa) inhibitors (argatroban, lepirudin, PPACK, enoxaparin, ZK-807834, fondaparinux) were investigated on the time course of thrombin catalytic activity triggered by the tissue factor pathway in platelet-poor plasma (PPP) of male healthy volunteers using the Calibrated Automated Thrombogram((R)) (CAT) method. In the presence of the anticoagulant drugs, the thrombin activity profiles were dose-dependently modified according to their specific enzyme inhibitory activity. ZK-807834 was the most potent drug for reducing the C(max) and the V(max) but also for prolonging the T(max). Lepirudin most efficiently delayed the lag time whereas enoxaparin was the most powerfully drug for diminishing the endogenous thrombin potential (ETP). In conclusion, the thrombin activity profile performed with the CAT method is a very rapid, suitable and reliable pharmacological tool for screening thrombin and/or FXa inhibitors whatever their inhibition mode. It consists of a powerful alternative for the classical PT clotting assay, especially regarding to the time course and the total amount of active thrombin generated. Last but not least, it provides insight into the mechanism of action of the compounds. PMID- 19124080 TI - The scale of divergence: a phylogenetic appraisal of intercontinental allopatric speciation in a passively dispersed freshwater zooplankton genus. AB - Molecular studies have enlightened our understanding of freshwater zooplankton biogeography, yet questions remain regarding the scale and commonality of geographic speciation. Here, we present a mtDNA-based phylogenetic hypothesis for 92 Daphnia species from all seven continents, with a focus on North and South America, Europe, and Australia, and use it to explore the frequency, scale, and geographical orientation of allopatric divergence events. Allopatric speciation can conservatively account for at least 42% of cladogenetic events among the species included in our study; most of these involve intercontinental splits. Closely related species pairs are concentrated in the circumarctic region and between northern and southern continents, aligned with bird migration routes, suggesting recent dispersal. By contrast, deeper phylogenetic patterns are consistent with vicariance scenarios linked to continental fragmentation. The possible reasons for the puzzling persistence of these ancient patterns in light of the eroding force of dispersal are considered. Our results demonstrate the high frequency and complex pattern of allopatric speciation in this ancient, passively dispersed genus. PMID- 19124081 TI - Social determination of sex in reef fishes. AB - Many fishes living in reef environments display remarkable flexibility in sexuality with social interactions determining their sex either during juvenile development or in adulthood. The evolutionary advantages of such flexibility are relatively well established. By contrast, the mechanisms by which social cues guide development of the sexual phenotype are less well understood. This paper reviews our understanding of these processes for some well-studied reef fish groups at the gonadal and neuroendocrine levels as well as proposing promising directions for future study. PMID- 19124082 TI - Specification and loss of melanocyte stem cells. AB - The melanocyte stem cells of the hair follicle provide an attractive system for the study of stem cells. The stem cells exist in an anatomically defined niche clearly separated from their differentiated progeny, differentiated progeny can be selected against by treatment with an inhibitor of Stem Cell Factor signaling, perturbations which affect survival and differentiation have clearly visible pigmentation phenotypes, and genetic mutations can impair or completely abolish this lineage without lethality. In mice several coat color mutants have been shown to have impaired specification or survival of melanocyte stem cells. Furthermore understanding of the normal regulation and behaviors of melanocytes and melanocyte stem cells will allow development of better strategies for cancer treatment. This article will review the discovery and behaviors of melanoctye stem cells as well as some aspects of melanocyte biology. PMID- 19124083 TI - Protein kinase A regulates production of virulence determinants by the entomopathogenic fungus, Metarhizium anisopliae. AB - Metarhizium anisopliae is a model system for studying insect fungal pathogenesis. The role of cAMP signal transduction in virulence was studied by disrupting the class I PKA catalytic subunit gene (MaPKA1). The PKA mutant (DeltaMaPKA1) showed reduced growth and greatly reduced virulence. PKA was dispensable for differentiation of infection structures (appressoria), but differentiation was delayed and the appressoria were defective because of reduced turgor pressure. DeltaMaPKA1 germinated at similar rates as the wild type in glucose and glycerol, but germination was delayed on alanine. Conidial adhesion and appressorium formation by DeltaMaPKA1 against a plastic surface was fully inhibited with glucose as sole nutrient source. Adhesion to plastic was not inhibited with glycerol or alanine, but appressorium formation was delayed. DeltaMaPKA1 showed reduced tolerance to the oxidative agent diamide, but not to H(2)O(2) and methyl viologen. Comparative transcriptome analysis of DeltaMaPKA1 and the wild type strain showed that PKA is responsible for up-regulating approximately one-third of the genes induced by insect cuticle, including subsets of those responsible for differentiation of appressoria and penetration pegs, cuticle degradation, nutrient acquisition, pH regulation, lipid synthesis, cell cycle control and the cytoskeleton. PKA was not however required for expression of toxin-producing genes. We conclude therefore that MaPKA1 is required for sensing host-related stimuli and transduction of these signals to regulate many infection processes. PMID- 19124084 TI - Csk-binding protein can regulate Lyn signals controlling cell morphology. AB - The Src family kinase Lyn is involved in differentiation signals emanating from activated erythropoietin (Epo) receptors, it interacts with COOH-terminal Src kinase-binding protein (Cbp), an adaptor protein that recruits negative regulators COOH-terminal Src kinase (Csk) and suppressor of cytokine signaling-1 (SOCS1). Lyn phosphorylates Cbp on several tyrosine residues, including Tyr314, which recruits Csk/SOCS1, as well as Tyr381 and Tyr409 that bind Lyns own SH2 domain. We show that Cbp alters not only the ability of erythroid cells to differentiate but also their colony morphology. Consequently, we detailed the ability of Cbp to interact with and influence Lyns ability to initiate changes in cellular architecture, which affect cell-cell and cell-substratum interactions. Over-expression of active Lyn promotes filopodia formation while inactive Lyn promotes lamellipodia formation. Conversely, Cbp over-expression, which inhibits Lyn activity, promotes lamellipodia formation, while Cbp mutants preventing its interaction/signaling consequently allow Lyn to promote filopodia formation. Thus, the Lyn-Cbp pathway and subsequent regulation of Lyn signaling and cell morphology involves a dynamic and complex series of interactions. PMID- 19124086 TI - A new evaluation methodology for literature-based discovery systems. AB - While medical researchers formulate new hypotheses to test, they need to identify connections to their work from other parts of the medical literature. However, the current volume of information has become a great barrier for this task. Recently, many literature-based discovery (LBD) systems have been developed to help researchers identify new knowledge that bridges gaps across distinct sections of the medical literature. Each LBD system uses different methods for mining the connections from text and ranking the identified connections, but none of the currently available LBD evaluation approaches can be used to compare the effectiveness of these methods. In this paper, we present an evaluation methodology for LBD systems that allows comparisons across different systems. We demonstrate the abilities of our evaluation methodology by using it to compare the performance of different correlation-mining and ranking approaches used by existing LBD systems. This evaluation methodology should help other researchers compare approaches, make informed algorithm choices, and ultimately help to improve the performance of LBD systems overall. PMID- 19124085 TI - Reassessing the amyloid cascade hypothesis of Alzheimer's disease. AB - Since its inception, the amyloid cascade hypothesis has dominated the field of Alzheimer's disease (AD) research and has provided the intellectual framework for therapeutic intervention. Although the details of the hypothesis continue to evolve, its core principle has remained essentially unaltered. It posits that the amyloid-beta peptides, derived from amyloid precursor protein (APP), are the root cause of AD. Substantial genetic and biochemical data support this view, and yet a number of findings also run contrary to its tenets. The presence of familial AD mutations in APP and presenilins, demonstration of Abeta toxicity, and studies in mouse models of AD all support the hypothesis, whereas the presence of Abeta plaques in normal individuals, the uncertain nature of the pathogenic Abeta species, and repeated disappointments with Abeta-centered therapeutic trials are inconsistent with the hypothesis. The current state of knowledge does not prove nor disprove the amyloid hypothesis, but rather points to the need for its reassessment. A view that Abeta is one of the factors, as opposed to the factor, that causes AD is more consistent with the present knowledge, and is more likely to promote comprehensive and effective therapeutic strategies. PMID- 19124087 TI - A measles-derived peptide treats and vaccinates against adjuvant arthritis. AB - Measles vaccine and porcine myelin basic protein were both found to ameliorate or abolish the symptoms of adjuvant arthritis (AA) in Lewis rats whether inoculated at the time of adjuvant administration or after the onset of arthritis. These results are consistent with clinical observations that measles infection can sometimes cause remission of juvenile rheumatoid arthritis. The fact that measles virus proteins and myelin basic protein have significant regions of homology allowed peptides based on these regions to be synthesized. A twenty-amino acid sequence exhibits significant anti-arthritic activity when inoculated into rats with pre-existing AA and it also prevented onset of AA when a single dose was preinoculated three weeks prior to AA induction. These data suggest the possibility of developing novel therapeutic vaccines against some forms of arthritis. PMID- 19124088 TI - Phospholipids influence the aggregation of recombinant ovine prions. From rapid extensive aggregation to amyloidogenic conversion. AB - The transformation of prion protein (PrP) into its insoluble amyloid form correlates with neurodegeneration in transmissible spongiform encephalopathies. PrP is connected to the neuronal membrane by a covalently-linked glycosylphosphatidylinositol (GPI) anchor. The current study demonstrates that phosphatidylinositol and phosphatidylethanolamine in low concentrations (0.5-50 muM) stimulate rapid unlimited aggregation of PrP. At a higher concentration (500 muM), lipid particles prevent the formation of large PrP aggregates and induce an increase in the beta-sheet structure content of PrP protein. Thus, the liberation of PrP from the membrane and its direct interaction with its own GPI moiety, as well as with membrane lipids, can promote the formation of aggregated structures of PrP. The phospholipids studied are also able to upregulate the aggregation of oligomeric PrP forms (12-mers and 36-mers), the neurotoxicity of which has been reported recently. Low phosphatidylinositol concentrations induce these oligomers to form aggregates of smaller size when compared with aggregates formed directly from monomers. The inhibition of extensive aggregation observed at a high concentration of phosphatidylinositol (500 muM) results in both the formation of amyloids from PrP monomers and the interaction of protein molecules with lipid micelles. Thus, phospholipids are not only involved in the aggregation of prion monomers and their amyloidogenic conversion, but also regulate the aggregative status of prion oligomers already formed. Consequently, depending on their micellar status, phospholipids can either promote amyloidogenic conversion and conserve neurotoxic oligomeric forms (lipid micelles) or mediate the formation of large-size amorphous aggregates (non-micellar phospholipids). PMID- 19124089 TI - Testosterone, but not IGF-1, LH, prolactin or cortisol, may serve as antler stimulating hormone in red deer stags (Cervus elaphus). AB - The role of androgens and insulin-like growth factor 1 (IGF-1) in antler growth has been disputed. We predicted that the secretory of IGF-1 may be associated with an acceleration of body growth rather than with antler growth. Furthermore we anticipated a relationship between the increase of testosterone and the progress of antler growth. If IGF-1 is involved in the stimulation of antler growth, this should be more obvious in young than in mature stags. Eight two-year old red deer stags (Cervus elaphus), and twelve adult red deer stags were blood sampled and the length of their velvet antlers was measured in one-week intervals during the period of antler growth. Concentrations of testosterone, cortisol, IGF 1, luteinizing hormone (LH), and prolactin were determined in plasma by enzyme immunoassay or radioimmunoassay. Antler growth per day was primarily dependent on changes in testosterone concentration per day in both groups of stags. As expected, only in two-year-old stags we detected a possible role of IGF-1 in the antler growth regulation, but that was not in agreement with previously published studies. Nevertheless, this effect was still utilized in interaction with testosterone. In addition to total antler length, only concentrations of testosterone and LH were significantly higher in adult males in comparison to two year-old males. Our present results lead us to conclude that it is not IGF-1 but testosterone which is responsible for the intensity of antler growth in subadult and adult red deer stags. PMID- 19124090 TI - Effects of cod bone gelatin on bone metabolism and bone microarchitecture in ovariectomized rats. AB - Several animal studies have showed that gelatin may be effective for minimizing bone loss in OVX rats with established osteopenia. To gain insight into how cod bone gelatin administration affects bone loss after ovariectomy, studies were carried out focusing on bone quality and the molecular mechanisms. Eighty-four female rats were ovariectomized, 12 sham-operated, divided into six groups of 12 each and treated one week after ovariectomy either with vehicle or cod bone gelatin (0.375, 0.75, 1.5, 3, 6 mg/kg body weight) for 90 days. Bone densitometry, microCT analysis, real-time PCR analysis and biochemical analysis were used at the end of the study. After 90 days, BMD of proximal tibia and femoral neck decreased in OVX rats, whereas the loss of BMD in those regions was prevented at 3 g/kg (P<0.05). However, the BMD of midshaft femurs showed no significant differences. BV/TV, Tb.N. and Tb.Th. in the 3 g/kg group were, respectively, 30.4% (P<0.05), 145.5% (P<0.05) and 81.5% (P<0.05) higher than in the OVX group. A significant decrease was detected in urine CTX, NTX and DPD, suggesting decreased bone resorption. Treatment with 3 g/kg and 6 g/kg cod bone gelatin attenuated the increase in serum IL-1beta, IL-6 and TNF-alpha observed in the OVX group. Real-time PCR showed significantly decreased levels of mRNA expression for RANKL at the dosage of 6 g/kg and the RANKL/OPG mRNA ratio in the 3 g/kg and 6 g/kg group significantly decreased compared to the OVX group (P<0.05). In conclusion, our data confirmed that the cod bone gelatin treatment at 3 g/kg is effective in the prevention of estrogen deficient bone loss by modulating the expression of RANKL and OPG and suppressing the release of proinflammatory cytokines. PMID- 19124093 TI - Retraction notice. PMID- 19124091 TI - Distribution of the alcohol dehydrogenase ADH1B*47His allele in Eurasia. PMID- 19124094 TI - Guest editors' introduction. Methods in tissue engineering. PMID- 19124095 TI - Breaking bad news in breast imaging: a critical component of patient care. PMID- 19124096 TI - 4-D Imaging in cerebrovascular disorders by using 320-slice CT: feasibility and preliminary clinical experience. AB - RATIONALE AND OBJECTIVES: The authors report study protocols and initial clinical experience in assessing patients with acute and chronic cerebrovascular disorders using the recently introduced technique of volume computed tomography (VCT). MATERIALS AND METHODS: Thirteen patients with presumptive cerebrovascular insufficiency underwent VCT using a 320-slice scanner (detector width, 160 mm), including time-resolved whole-brain perfusion and cerebral angiography (four dimensional computed tomographic angiography [CTA] and computed tomographic perfusion [CTP]). Unenhanced cranial CT (cCT) and helical cervicocranial CT (three-dimensional CTA) were added according to clinical requirements. Study protocols are presented, and image quality, data management, and radiation exposure issues are discussed. RESULTS: In 12 of 13 patients, the procedure was performed successfully on admission; in the other patient, the study was aborted for clinical reasons and repeated. Total scan time amounted to about 5 minutes, and data reconstruction times were up to 10 minutes. About 9000 primary images were generated, partially in the enhanced Digital Imaging and Communications in Medicine format, thus requiring new data postprocessing and management strategies. Image artifacts restricted the use of single-rotation cCT and incremental VCT (three-dimensional CTA). Overall exposure figures (computed tomographic dose index and dose-length product) were increased by 65% on average when three-dimensional CTA was added to volume cCT and four-dimensional CTA and CTP (5.0 mSv and 2178 mGy . cm, respectively). CONCLUSION: Preliminary clinical experience indicates that whole-brain four-dimensional CTA and CTP is a robust technique that provides relevant clinical information with respect to whole-brain perfusion as well as cerebral hemodynamics. The exposure benefit of deriving time resolved perfusion and vessel images from one source data set is compromised when adding three-dimensional CTA to the protocol. Other acquisition techniques specific to VCT, such as single-rotation cCT and incremental three-dimensional CTA, suffer from restrictions in terms of image quality at present. PMID- 19124097 TI - Breaking bad news in the breast imaging setting. AB - RATIONALE AND OBJECTIVES: The aim of this study was to investigate the experience and desire for training breast imagers in discussing "bad news" with their patients. No such information has been previously reported. MATERIALS AND METHODS: Following University of Michigan (UM) institutional review board approval and approval from the Society of Breast Imaging (SBI), questionnaires were mailed to current UM breast imaging faculty members, fellows, and residents and to SBI fellows. The final questionnaire page was mistakenly omitted in the SBI mailing. After institutional review board approval, the final page was sent to the SBI fellows. RESULTS: The response rates were 65% (61 of 94) for SBI fellows and 65% (13 of 20) for UM physicians. Ninety-five percent of respondents were aged >or= 40 years. Sixty-two percent of UM physicians were women, compared to 52% of SBI fellows. Sixty-four percent of respondents practiced in university settings and 34% in private settings. Thirty-five percent of respondents had breast imaging fellowships. The frequency of delivering bad news was most often weekly or daily, whether the news was a need for extra views, a biopsy recommendation, or positive core biopsy results. Recommendations for extra views and breast biopsies were reported to be usually given in person, whereas 59% of respondents delivered positive biopsy results by telephone. Eighty-five percent of respondents noted no previous training in delivering bad news. On a comfort scale ranging from 1 (very uncomfortable) to 10 (very comfortable), 95% of respondents rated their comfort at 8 to 10 for recommending extra views, 85% had the same comfort level for recommending biopsies, and 67% had the same level for giving positive biopsy results. Sixty-two percent of UM radiologists expressed interest in obtaining further training in how to deliver bad news, whereas only 32% of SBI fellows were desirous of such training. CONCLUSION: Although radiologists commonly delivered bad news in the breast imaging setting, very few had training in how to do so. Comfort levels declined as the severity of news increased, yet a majority of respondents were not interested in obtaining training in how to deliver bad news. These findings raise the need for further study. PMID- 19124098 TI - Prophylactic vertebroplasty: cement injection into non-fractured vertebral bodies during percutaneous vertebroplasty. AB - RATIONALE AND OBJECTIVES: We investigated the efficacy of prophylactic cement injection into the vertebral body adjacent to fractured vertebra to prevent new fractures after percutaneous vertebroplasty (PV). MATERIALS AND METHODS: Between February 2002 to August 2004, PV was performed for osteoporotic compression fractures in 89 consecutive patients. All patients underwent PV for only fractured vertebrae. Between September 2004 and October 2006, we performed prophylactic cement injection for 155 patients, with cement injected into the non fractured vertebra adjacent to the fractured vertebra, immediately above the fractured vertebra in the same procedure. We evaluated the frequency of new vertebral fractures and the efficacy of prophylactic therapy. RESULTS: In the non prophylactic group, 15 of 89 patients (16.8%) developed new fractures within 3 months and 20 of 89 patients (22.4%) developed new painful compression fractures within a year after the first PV. These fractures occurred mostly in adjacent vertebra, particularly in the vertebra immediately superior to the treated one and occurred in the lower thoracic and upper lumbar spine. In the prophylactic group, 7 of 155 patients (4.5%) developed new compression fractures within 3 months and 15 of 155 patients (9.7%) developed new compression fractures within 1 year. Statistical analysis showed that fewer new fractures developed in the prophylactic group than in the non-prophylactic group at both 3 months (P = .0020, Fisher's exact test) and 1 year (P = .0079). CONCLUSIONS: Prophylactic cement injection into non-fractured vertebrae adjacent to fractured vertebrae may prevent new compression fractures after vertebroplasty for osteoporotic patients. PMID- 19124099 TI - Contrast enhancement in chest multidetector computed tomography: intraindividual comparison of 300 mg/ml versus 400 mg/ml iodinated contrast medium. AB - RATIONALE AND OBJECTIVES: We sought to intraindividually compare intravascular contrast enhancement in multidector computed tomography (MDCT) of the chest using contrast media (CM) containing 300 and 400 mg iodine/ml. MATERIALS AND METHODS: Seventy-one patients underwent repeated MDCT scanning of the chest at baseline and follow-up. CM with standard iodine (protocol A: 300 mg iodine/ml; Iopromide 300) and high iodine concentration (protocol B: 400 mg iodine/ml; Iomeprol 400) were used. The iodine delivery rate (1.29 g iodine/s) and total iodine load (37 g iodine) were identical for the two protocols. Contrast enhancement was measured in the right and left ventricles, pulmonary trunk, right and left pulmonary arteries, and ascending and descending aortas. Results were compared using paired t-tests; P values were adjusted using Bonferroni correction (P .005). In the right ventricle, pulmonary trunk, and right and left pulmonary arteries, higher attenuation values for protocol A were detected compared to protocol B (379.0 +/- 110.5 vs. 349.8 +/- 117.6, 354.5 +/- 112.2 vs 330.9 +/- 118.3, 348.6 +/- 106.0 vs. 321.8 +/- 109.9, and 347.9 +/- 102.4 vs. 321.0 +/- 104.9 HU, respectively). After the lung circulation (left ventricle, ascending aorta, and descending aorta), attenuation values were marginally higher for protocol B. Using both protocols resulted in suitable contrast enhancement with a mean pulmonary attenuation higher than 300 HU. CONCLUSIONS: Using an adapted injection protocol, the administration of 300 and 400 mg iodine CM resulted in a suitable intravascular contrast enhancement in the chest. The use of 400 mg iodine CM does not lead to a statistically significant improvement in contrast enhancement compared to the 300 mg iodine CM. PMID- 19124100 TI - Helical multidetector row quantitative computed tomography (QCT) precision. AB - RATIONALE AND OBJECTIVES: The impact of varying image acquisition parameters on the precision of measurements using quantitative computed tomography is currently based on studies performed before the advent of helical image acquisition and multidetector-row scanners. The aim of this study was to evaluate helical multidetector-row quantitative computed tomography to determine the factors contributing to the overall precision of measurements on quantitative computed tomography conducted using current vintage computed tomographic (CT) scanners. MATERIALS AND METHODS: The effects of CT protocol parameters (x-ray tube voltage and current, pitch, gantry rotation speed, detector configuration, table height, and reconstruction algorithm) and short-term scanner variation were examined on two commercially available quantitative CT (QCT) systems (ie, a combination of reference phantoms and analysis software) using seven multidetector-row CT scanners (available from a single vendor) operated in helical mode. Combined with simulated patient repositioning using three ex vivo spine specimens, precision (coefficient of variation) estimates were made on the basis of three scenarios: "best case," "routine case," and "worst case." RESULTS: The overall best-case QCT precision was 1.4%, provided that no changes were permitted to the bone mineral density (BMD) scan protocol. Routine-case examination (with a BMD reference phantom in place) that permitted some variation in the x-ray tube current and table speed produced a precision of 1.8%. Without any constraints on the clinical QCT examinations, the worst-case precision was estimated at 3.6%. CONCLUSIONS: Although small in appearance, these errors are for single time points and may increase substantially when monitoring changes through QCT measurements over several time points. This calls for increased caution and attention to detail whenever using helical multidetector-row quantitative computed tomography for the assessment of BMD change. PMID- 19124101 TI - Dual-energy CT discrimination of iodine and calcium: experimental results and implications for lower extremity CT angiography. AB - RATIONALE AND OBJECTIVES: The purpose of this work was to measure the accuracy of dual-energy computed tomography for identifying iodine and calcium and to determine the effects of calcium suppression in phantoms and lower-extremity computed tomographic (CT) angiographic data sets. MATERIALS AND METHODS: Using a three-material basis decomposition method for 80- and 140-kVp data, the accuracy of correctly identified contrast medium and calcium voxels and the mean attenuation before and after calcium suppression were computed. Experiments were first performed on a phantom of homogenous contrast medium and hydroxyapatite samples with mean attenuation of 57.2, 126, and 274 Hounsfield units (HU) and 50.0, 122, and 265 HU, respectively. Experiments were repeated in corresponding attenuation groups of voxels from manually segmented bones and contrast medium enhanced arteries in a lower-extremity CT angiographic data set with mean attenuation of 293 and 434 HU, respectively. Calcium suppression in atherosclerotic plaques of a cadaveric specimen was also studied, using micro computed tomography as a reference, and in a lower-extremity CT angiographic data set with substantial below-knee calcified plaques. RESULTS: Higher concentrations showed increased accuracy of iodine and hydroxyapatite identification of 87.4%, 99.7%, and 99.9% and 88.0%, 95.0%, and 99.9%, respectively. Calcium suppression was also more accurate with higher concentrations of iodine and hydroxyapatite, with mean attenuation after suppression of 47.1, 122, and 263 HU and 7.14, 11.6, and 12.6 HU, respectively. Similar patterns were seen in the corresponding attenuation groups of the contrast medium-enhanced arteries and bone in the clinical data set, which had overall accuracy of 81.3% and 78.9%, respectively, and mean attenuation after calcium suppression of 254 and 73.7 HU, respectively. The suppression of calcified atherosclerotic plaque was accurate compared with the micro-CT reference; however, the suppression in the clinical data set showed probable inappropriate suppression of the small vessels. CONCLUSION: Dual-energy computed tomography can detect and differentiate between contrast medium and calcified tissues, but its accuracy is dependent on the CT density of tissues and limited when CT attenuation is low. PMID- 19124102 TI - Computer-aided detection of endobronchial valves using volumetric CT. AB - RATIONALE AND OBJECTIVES: The ability to automatically detect and monitor implanted devices may serve an important role in patient care by aiding the evaluation of device and treatment efficacy. The purpose of this research was to develop a system for the automated detection of one-way endobronchial valves that were implanted for less invasive lung volume reduction. MATERIALS AND METHODS: Volumetric thin-section computed tomographic data was obtained for 194 subjects; 95 subjects implanted with 246 devices were used for system development and 99 subjects implanted with 354 devices were reserved for testing. The detection process consisted of preprocessing, pattern recognition based detection, and a final device selection. Following the preprocessing, a set of classifiers was trained using AdaBoost to discriminate true devices from false positives. The classifiers in the cascade used two simple features (either the mean or maximum attenuation) of a local region computed at multiple fixed landmarks relative to a template model of the valve. RESULTS: Free-response receiver-operating characteristic analysis was performed for the evaluation; the system could be set so the mean sensitivity was 96.5% with a mean of 0.18 false positives per subject. If knowledge of the number of implanted devices were incorporated, the sensitivity would be 96.9% with a mean of 0.061 false positives per subject; this corresponds to a total of 12 false negatives and six false positives for the 99 subjects in the test dataset. CONCLUSION: Software was developed for automated detection of endobronchial valves on volumetric computed tomography. The proposed device modeling and detection techniques may be applicable to other devices as well as useful for evaluation of treatment response. PMID- 19124103 TI - Comparison of the accuracy of CT volume calculated by circumscription to prolate ellipsoid volume (bidimensional measurement multiplied by coronal long axis). AB - RATIONALE AND OBJECTIVES: Tumor volume is one of the most important factors in evaluating the response to treatment of patients with cancer. The objective of this study was to compare computed tomographic (CT) volume calculation using a semiautomated circumscribing tracing tool (manual circumscription [MC]) to prolate ellipsoid volume calculation (PEVC; bidimensional measurement multiplied by coronal long axis) and determine which was more accurate and consistent. MATERIALS AND METHODS: The study included six patients with nine neoplasms, six phantoms, and two radiologists. The neoplasms and phantoms of varying sizes and shapes were imaged using multidetector CT scanners, with slice thicknesses ranging from 0.5 to 3 mm. Measurements were performed using a TeraRecon 3D workstation. Each lesion and phantom was manually circumscribed, and its three dimensions were measured. The measurements were repeated 2 weeks later. RESULTS: MC of the phantoms deviated from their true volumes by an average of 3.0 +/- 1%, whereas PEVC deviated by 10.1 +/- 3.99%. MC interobserver readings varied by 1.2 +/- 0.6% and PEVC by 4.8 +/- 3.3%. MC intraobserver readings varied by 1.95 +/- 1.75% and PEVC by 2.5 +/- 1.55%. Patient tumor volume predicted by MC and PEVC varied greatly; MC interobserver readings differed by 3.3 +/- 2.1% and PEVC by 20.1 +/- 10.6%. MC intraobserver readings varied by 2.5 +/- 1.9% and PEVC by 5.5 +/- 3.2%. Variability was greater for complex shapes than for simple shapes. Bidimensional analysis demonstrated an interobserver difference of 12.1 +/- 8.7% and an intraobserver difference of 5.05 +/- 3.3%. These results demonstrate large interobserver and intraobserver variability. Variability was greater for complex shapes than for simple shapes. CONCLUSION: MC of neoplasms provided more accurate and consistent volume predictions than PEVC. More complicated shapes demonstrated the superiority of MC over PEVC. PMID- 19124104 TI - Coronary artery calcium scoring on low-dose prospective electrocardiographically triggered 64-slice CT. AB - RATIONALE AND OBJECTIVES: The purpose of this prospective study was to assess image noise and variability in repeated coronary artery calcium (CAC) scoring on low-dose prospective electrocardiographically-triggered 64-slice multidetector computed tomography. MATERIALS AND METHODS: Patients (n = 115) suspected of having coronary artery disease were scanned twice, using a tube current of 10 x body mass index mA. The standard deviation (SD) of the computed tomographic value in the ascending aorta and (mean + 2 x SD) were obtained. Repeated CAC scores (Agatston, volume, and mass) were measured by two observers, and the interscan and interobserver variability were determined. RESULTS: The mean tube current used was 246 +/- 36 mA. The mean tube current-time product and mean estimated effective dose were 57 +/- 8 mA and 0.9 +/- 0.2 mSv, respectively. The SD and (mean + 2 x SD) computed tomographic values in the ascending aorta were 16 +/- 3 and 75 +/- 10 Hounsfield units, respectively. Repeated CAC scores were correlated (r(2) = 0.995-0.998). The interscan variability for observer 1 and observer 2, respectively, were 13% and 13% for Agatston score, 12% and 11% for volume, and 11% and 11% for mass. The interobserver variability for scan 1 and scan 2, respectively, were 3% and 3% for Agatston score, 5% and 3% for volume, and 3% and 3% for mass. CONCLUSION: Low-dose prospective electrocardiographically-triggered 64-slice multidetector computed tomography shows low interscan and interobserver variability on CAC scoring while maintaining low image noise. PMID- 19124105 TI - Progression of coronary atherosclerosis after heart transplantation on electron beam computed tomography. AB - RATIONALE AND OBJECTIVES: Cardiac transplant vasculopathy is the most important long-term complication of heart transplantation, with overlapping features with conventional, atherosclerotic coronary artery disease. The aim of this study was to determine the progression of coronary artery disease after heart transplantation by measuring total coronary calcium load. MATERIALS AND METHODS: After heart transplantation, 185 patients were serially examined using electron beam computed tomography for coronary calcium load for clinical reasons. The mean time between the initial examination and the follow-up scan was 566 days (range, 126-1,436). Coronary calcium load was measured by the Agatston method, and the total calcium scores at both examinations were compared between patients taking and those not taking lipid-lowering medications (statins). RESULTS: Patients not taking statins (n = 94) displayed a median annualized percentage increase in total calcium score of 0 Agatston units, whereas patients taking at least the lowest recommended daily dose of a statin (n = 84) displayed an annualized percentage decrease of 11 Agatston units. The difference was not statistically significant (Wilcoxon's rank-sum test, P = .35). Only 17 patients had increases of > 24 Agatston units, and eight of them were taking statins (chi(2) test, P = .99). CONCLUSION: The annual rate of progression of coronary calcium load after heart transplantation is low. In this investigation, no beneficial effects of statins could be detected. PMID- 19124106 TI - Quantitative measurement of iodine concentration in the liver using abdominal C arm computed tomography. AB - RATIONALE AND OBJECTIVES: C-arm computed tomography is an option on a C-arm angiographic system capable of acquiring projections while rotating the C-arm around the patient and reconstructing cross-sectional images with improved contrast resolution of 5 to 10 Hounsfield units. Typical abdominal C-arm computed tomographic (CCT) images, however, exhibit artifacts with spatially varying and drifting pixel values. Considering liver tumor oncologic procedures, the aim of this study was to evaluate the accuracy of liver iodine concentration (IC) estimated from CCT images under such challenging conditions. MATERIALS AND METHODS: The proposed method estimates the IC in a region of interest (ROI) using pixel values of CCT images measured at the ROI and a nonenhanced background. Two approaches to measure the background value were tested: one approach, L-BG, measured a corresponding local background value near each ROI, and the other, G BG, used one global background value for the entire object. The accuracy of estimations using CCT and computed tomographic scanners was evaluated; an elliptical cylinder water phantom with iodine solution inserts and seven patient data sets with transcatheter arterial chemoembolization were used. RESULTS: With the least "truncation" (the edge of the object being outside the field of view) of 27 mm, the IC was accurately estimated with CCT images (n = 9; root-mean square error [RMSE], 1.60-1.63 mg/mL; normalized RMSE, 11.8%; r(2) = 0.97; P < .001), with the true concentration ranging from 2.32 to 31.82 mg/mL. With truncations of up to 100 mm (n = 88), the estimation by L-BG remained accurate independent of the amount of truncation (RMSE, 1.58 mg/mL; normalized RMSE, 12.5%; r(2) = 0.06; P = .02), whereas the estimation by G-BG reduced the accuracy (RMSE, 4.61 mg/mL; normalized RMSE, 34.3%; r(2) = 0.10; P = .003). Clinical data (n = 37) showed that the estimation from CCT images using the L-BG method agreed well with that from computed tomographic images (RMSE, 2.86 mg/mL; normalized RMSE, 38.7%; r(2) = 0.76; P < .001). CONCLUSION: The liver IC can be accurately estimated with abdominal CCT images. PMID- 19124107 TI - Reducing the number of reader interpretations in MRMC studies. AB - RATIONALE AND OBJECTIVES: Multireader, multicase (MRMC) receiver-operating characteristic studies often require large numbers of patients, readers, and reader interpretations. The objective of this work is to evaluate a new "mixed" MRMC study design that reduces the number of reader interpretations. MATERIALS AND METHODS: As compared to the traditional MRMC design, the number of reader interpretations and the number of cases that must be truth-verified for the new mixed design was evaluated theoretically and empirically for various correlation values and sample sizes. RESULTS: For large MRMC studies, the new mixed design offers a substantial savings in the number of reader interpretations if the magnitude of the difference in between-reader correlations is not zero. For example, compared to a traditional design with 20 readers, 200 total cases, and a difference in between-reader correlations of 0.05, the newly proposed mixed design requires each reader to interpret only 132 cases, but at a cost of truth verifying an additional 64 cases. CONCLUSIONS: The mixed design can reduce the number of cases that readers need to interpret and the overall duration of a study, but at a cost in terms of the number of cases that must be truth-verified. The mixed design is particularly useful for studies where the condition being detected is not rare and patients routinely undergo the gold standard assessment. PMID- 19124108 TI - Prediction of metastatic disease and survival in patients with gastric and gastroesophageal junction tumors: the incremental value of PET-CT over PET and the clinical role of primary tumor volume measurements. AB - RATIONALE AND OBJECTIVES: To investigate the accuracy of M staging (staging of metastatic disease) in esophageal carcinoma based on a visual interpretation and based on tumor volume measurements on positron emission tomography (PET) computed tomography (CT). MATERIALS AND METHODS: Fifty-nine untreated patients with gastroesophageal junction tumors were enrolled, including 36 subcardial gastric tumors (type III according to Siewert classification) and 23 adenocarcinomas of the cardia (AEG, type II Siewert). Patients were grouped in metastasis free (M0 stage, n = 34) and metastatic stages (M1 stage, n = 25). Tumor volume and mean and maximum standardized uptake value were measured on PET-CT. The accuracy of these quantitative tumor volume parameters in distinguishing metastasis-free tumors (M0 stage) from metastatic stages (M1 stage) was compared to the accuracy of a visual analysis with fused PET-CT. Furthermore, accuracy of PET-CT was compared to PET reviewed side by side with CT in a lesion-based analysis of 84 distant metastatic sites. RESULTS: In the visual interpretation, PET-CT (accuracy 88%, 74/84) was more accurate than PET (accuracy 78%, 66/84; P = .008) in characterizing the 84 potential metastatic sites in the 59 patients. Among the tumor parameters, the PET-CT tumor volume was the most accurate predictor of M1 stage and overall survival. With a threshold of 39 mL, PET-CT volume was able to predict M1 stage disease with a sensitivity of 96% and a specificity of 85%. The accuracy of M-staging was increased further when combining tumor volume measurements with the results from the visual analysis (combined results: sensitivity 96%, specificity 94%). CONCLUSIONS: PET-CT was more accurate than PET (reviewed side by side with CT) in characterizing distant metastatic sites of gastroesophageal junction carcinomas. The highest accuracy for M-staging was obtained when combining the results of the visual analysis with the results from primary tumor volume measurements. Primary tumor volume was shown to be an independent prognostic factor. PMID- 19124109 TI - Statistical approaches for modeling radiologists' interpretive performance. AB - Although much research has been conducted to understand the influence of interpretive volume on radiologists' performance of mammography interpretation, the published literature has been unable to achieve consensus on the volume standards required for optimal mammography accuracy. One potential contributing factor is that studies have used different statistical approaches to address the same underlying scientific question. Such studies have relied on multiple mammography interpretations from a sample of radiologists; thus, an important statistical issue is appropriately accounting for dependence, or correlation, among interpretations made by (or clustered within) the same radiologist. The aim of this review is to increase awareness about differences between statistical approaches used to analyze clustered data. Statistical frameworks commonly used to model binary measures of interpretive performance are reviewed, focusing on two broad classes of regression frameworks: marginal and conditional models. Although both frameworks account for dependence in clustered data, the interpretations of their parameters differ; hence, the choice of statistical framework may (implicitly) dictate the scientific question being addressed. Additional statistical issues that influence estimation and inference are also discussed, together with their potential impact on the scientific interpretation of the analysis. This work was motivated by ongoing research being conducted by the National Cancer Institute's Breast Cancer Surveillance Consortium; however, the ideas are relevant to a broad range of settings in which researchers seek to identify and understand sources of variability in clustered binary outcomes. PMID- 19124110 TI - Determining optimal acquisition parameters for computed tomography coronary angiography: evaluation of a software-assisted, breathhold exam simulation. AB - RATIONALE AND OBJECTIVES: Scanning parameters used in multislice computed tomographic (MSCT) coronary angiography should be adapted to patients' heart rates. The aim of this study was to evaluate the rate of success of a software assisted scan simulation to determine optimal gantry rotation time prior to MSCT coronary angiography. MATERIALS AND METHODS: Data from 218 64-slice MSCT coronary angiographic studies were retrospectively analyzed. Prior to the MSCT examination, a scan had been simulated by giving a breath-hold command, after which a software program recorded the patient's heart rate from an electrocardiogram over the expected scanning time and predicted optimal scanning parameters. The success rate in predicting optimal parameters for the subsequent MSCT examination and the resulting acquisition window width were analyzed. RESULTS: There was a wide range of heart rates among the patients. The mean intraindividual variation during scan simulation and examination was 4.8 beats/min. Optimal scan parameters were selected in 179 of 218 cases (82%). The median acquisition window width achieved in this group was 174 ms (range, 100 200), compared with 192 ms (range, 149-225) in cases in which suboptimal settings were selected (P < .001). Correct prediction occurred significantly more often in patients with low heart rates (<66 beats/min) than in those with higher heart rates (>or=66 beats/min) (93% vs 66%; P < .001). CONCLUSIONS: The software assisted scan simulation was a fast and simple procedure that allowed the selection of optimal computed tomographic parameters in >80% of patients. The procedure may be a useful adjunct to avoid unwanted synchronicity of gantry rotation and heart cycle and optimize temporal resolution in MSCT coronary angiography. PMID- 19124111 TI - Colonoscopies, real or virtual. PMID- 19124113 TI - Prevalence, characteristics, and impact of bloating symptoms in patients with irritable bowel syndrome. AB - BACKGROUND & AIMS: Bloating symptoms are common in patients with irritable bowel syndrome (IBS) seen in primary care and gastrointestinal clinics. However, the underlying mechanisms of IBS are poorly understood, and there are few data available about the epidemiology of this syndrome or the impact of its symptoms. We investigated the prevalence, characteristics, and impact of bloating symptoms in patients with IBS. METHODS: IBS patients were identified by Rome II criteria in a U.S. population representative web-based survey. Patients were asked about the quality, frequency, and severity of their gastrointestinal symptoms. The impact of these symptoms was investigated by assessing patients' health-related quality of life, utilization of health care, and use of medications. RESULTS: Of the 337 IBS patients in this study, 82.5% (n = 278) reported bloating symptoms, the second most bothersome symptom after abdominal cramping. The symptoms were more prevalent in female patients, 87.4% (n = 209), than in male patients, 70.4% (n = 69) (P < .0001), and in patients with constipation, 88.7% (n = 47), and mixed symptoms, 88.8% (n = 135), than in patients with diarrhea, 72.3% (n = 96), (P = .02 and P < .01, respectively). Bloating symptoms were the third (of 14) most important reason to seek medical care, and more than half of the patients reported regular use of anti-gas medications. Bloating symptoms were associated with decreased energy levels (P = .04), food intake (P < .01), and physical functioning (P = .06). CONCLUSIONS: Bloating symptoms are common in patients with IBS, and their prevalence and relative severity differ on the basis of sex and IBS subtype. Bloating symptoms are associated with a decrease in the quality of life and increases in health care utilization and use of medications. PMID- 19124114 TI - Irritable bowel syndrome: more than abdominal pain and bowel habit abnormalities. AB - BACKGROUND & AIMS: Although irritable bowel syndrome (IBS) can be defined using few symptoms, principal symptoms alone may be inadequate in monitoring disorder severity. Secondary analysis of a published data set was performed to determine if more inclusive symptom measures would better reflect the burden of this disorder. METHODS: From a prospective naturalistic study of 213 patients meeting Rome II criteria, all the data were used from daily questionnaires recorded for 4 weeks, and repeated again after an interval of 4 weeks. The total number of 11 symptoms and intensity grading score of each symptom were analyzed alongside individual symptom intensities by principal component analysis. RESULTS: The trend accounting for the most variance was explained by the intensity of all symptoms together. The second largest trend was explained by differences between IBS bowel habits (constipation and diarrhea). The 2 constipation and 4 diarrhea symptoms closely correlated within each group, but the category of other symptoms were not correlated directly with either, and represent a separate dimension. Other symptoms (pain/discomfort, abdominal uneasiness, flatulence/distension, incomplete evacuation, pain or burning in the stomach) correlated more highly with disease intensity than either constipation or diarrhea symptoms. The sum of all symptoms and their intensity was consistent over each week, although the relative intensity of individual symptoms was more variable. Investigator measures of disease intensity underestimated that reported by patients. CONCLUSIONS: Non-bowel habit symptoms include more than abdominal pain and discomfort, and contribute to the largest component of the total symptom burden. Thus, more than bowel habits and abdominal pain drive IBS symptom severity. PMID- 19124115 TI - Increased cyclooxygenase-2 expression in juvenile polyposis syndrome. AB - BACKGROUND & AIMS: Gastrointestinal juvenile polyps may occur in juvenile polyposis syndrome (JPS) or sporadically. JPS is an autosomal-dominant condition caused by a germline defect in SMAD4 or BMPR1A in 50% to 60% of cases, and is characterized by multiple juvenile polyps, predominantly in the colorectum. JPS has an increased risk of gastrointestinal malignancy but sporadic juvenile polyps do not. Cyclooxygenase-2 (COX-2) expression is increased in gastrointestinal tumorigenesis and familial adenomatous polyposis. Inhibition of COX-2 leads to regression of colorectal adenomas in familial adenomatous polyposis patients and inhibits gastrointestinal tumorigenesis. To investigate the role of COX-2 in juvenile polyps, we compared the expression of COX-2 in juvenile polyps from a well-defined group of juvenile polyposis patients and sporadic juvenile polyps. METHODS: COX-2 expression was assessed in 24 genetically well-defined JPS patients and 26 patients with sporadic juvenile polyps using tissue microarray analysis. Two additional markers, Hu-antigen R, a stabilizer of messenger RNA, and CCAAT/enhancer-binding protein beta, a transcription factor, both associated with increased COX-2 expression, also were investigated. RESULTS: Increased COX-2 expression in JPS patients was noted compared with patients with sporadic juvenile polyps (P < .001). Also, JPS patients with a BMPR1A germline defect had higher COX-2 expression than did JPS patients in whom no germline mutation was detected. High COX-2 levels correlated with increased cytoplasmic Hu-antigen R expression in JPS polyps (P = .022), but not in sporadic juvenile polyps. CONCLUSIONS: Juvenile polyposis and sporadic juvenile polyps show distinctive expression profiles of COX-2 that may have clinical implications. PMID- 19124116 TI - Food hypersensitivity as a cause of rectal bleeding in adults. AB - BACKGROUND & AIMS: Rectal bleeding and lymphonodular hyperplasia (LNH) in children can be caused by food hypersensitivity (FH). Our aim was to verify whether similar clinical and endoscopy presentations in adults can be due to FH. METHODS: Consecutive adult patients with rectal bleeding were enrolled. All underwent routine assays, colonoscopy, and histology study. RESULTS: Ten of 64 (15%) patients showed LNH as the unique sign at colonoscopy. An oligoantigenic diet resolved the rectal bleeding in 9 patients, and the reintroduction of several foods caused symptom reappearance. Double-blind placebo-controlled challenges with cow's milk and wheat protein confirmed the FH; symptoms reappeared 1-96 hours after the challenge. None of the patients were positive for IgE-mediated assays. In patients with LNH and FH, histology of the ileum and colon mucosa showed a higher number of lymphoid follicles and intraepithelial and lamina propria eosinophils compared with the other patients with rectal bleeding. CONCLUSIONS: Recurrent rectal bleeding can be caused by FH in adult patients. Endoscopic evidence of LNH characterizes these cases. PMID- 19124117 TI - Visual objects can potentiate a grasping neural simulation which interferes with manual response execution. AB - Previous studies on visuomotor priming have provided insufficient information to determine whether the reach-to-grasp potentiation of a non-target object produces a specific effect during response execution. In order to answer this question, subjects were instructed to reach and grasp a response device with either a power or a precision grip, depending on whether the stimulus they saw was empty or full. Stimuli consisted of containers (graspable with either a power or a precision grip), with non-graspable stimuli added as a control condition (geometrical shapes). The image of the non-target object was removed during the execution phase. Results demonstrate slower execution responses related to motor incompatibility, though conversely, no faster responses with motor compatibility. Moreover, any visuomotor priming effect required that the container be displayed during response execution. These data suggest that during response execution, motor incompatibility produces a disruptive effect likely due to competition between two cerebral events: motor control of the actual response execution and visual object reach-to-grasp neural simulation. PMID- 19124118 TI - In vitro antimicrobial effect of bacteriocin PsVP-10 in combination with chlorhexidine and triclosan against Streptococcus mutans and Streptococcus sobrinus strains. AB - OBJECTIVES: To determine the minimal inhibitory concentrations (MICs) of bacteriocin PsVP-10, chlorhexidine and triclosan on S. mutans and S. sobrinus and to study the potential synergistic combination between these antimicrobial and the bacteriocin PsVP-10. DESIGN: Were determined MICs of bacteriocin PsVP-10, triclosan and chlorhexidine on strains of S. mutans and S. sobrinus, which formed a biofilm or did not form a biofilm. In addition, the synergistic effect was analysed by the determination of respective fractionary inhibitory concentrations (FICs) between bacteriocin PsVP-10 plus chlorhexidine and bacteriocin PsVP-10 plus triclosan. RESULTS: MICs of three antimicrobials used were higher in those bacterial strains, which formed a biofilm. An interesting synergistic effect on both studied species was observed when bacteriocin and chlorhexidine were combined. A slighter synergy was determined for the combination bacteriocin PsVP 10 and triclosan. CONCLUSIONS: The results showed that the combination of chlorhexidine bacteriocin PsVP-10 could reduce the number of cariogenic bacteria for in vitro studies. In the future this synergistic combination could be an alternative to antimicrobial therapy against S. mutans or S. sobrinus. PMID- 19124119 TI - Dysfunctional schema modes, childhood trauma and dissociation in borderline personality disorder. AB - OBJECTIVE: To examine the relationship between dysfunctional schema modes, childhood trauma and dissociation in borderline personality disorder (BPD). METHOD: 30 BPD patients completed the Wessex Dissociation Scale (WDS), Childhood Trauma Questionnaire (CTQ), General Health Questionnaire (GHQ), and Schema Mode Questionnaire (SMQ). RESULTS: The 'Angry and Impulsive Child' and 'Abandoned and Abused Child' modes uniquely predicted dissociation scores. Childhood trauma did not predict dissociation scores. CONCLUSIONS: Findings support the schema mode model of BPD [Young, J. E., Klosko, J. S., & Weishaar, M. E. (2003). Schema therapy: A practitioners guide. London: Guilford Press] and its emphasis on the role of dissociation. Clinically they support the emphasis on the identification and integration of dysfunctional parts of the personality in working with individuals diagnosed with BPD. PMID- 19124120 TI - Bone remodeling regulation under unloading conditions: numerical investigations. AB - The present paper addresses the following question: can a simple regulatory bone remodeling model predict effects of unloading conditions on the trabecular bone morphology? In an attempt to answer this question, rat tail-suspension was chosen as a model that mimics the microgravity environment. Over 23 days, histomorphometric analysis was carried out on cross-sections of tibias of the suspended animals. The slices were digitalized and images discretized to obtain osteocyte distribution and apparent bone density. Based on these experimental data, finite element simulations were conducted to evaluate the bone loss and the change in trabecular architecture similar to those observed after a spaceflight. The numerical model is driven by a remodeling law that takes into account the nonuniform osteocyte distribution that may itself provide mechanoreception. We used the bone density rate of change from the remodeling theory and a time stepping algorithm witch are implemented in a finite element software. This approach takes into account the unloading effects on bone remodeling process and permits to confront experimental and numerical data. We showed that there is a good agreement between these data, particularly at the beginning of the simulated bone mass loss during the rat tail-suspension experiment. Indeed, we obtained a variation of 5.25% at day 7 (D7), 2.09% at day 13 (D13) and finally, 51.03% at day 23 (D23). Despite that last variation, the proposed theoretical model can be suitable to simulate the alteration of bone mineral density under the specific unloading conditions of the rat tail-suspension model. PMID- 19124121 TI - Effect of simvastatin on kidney function loss in patients with coronary heart disease: findings from the Scandinavian Simvastatin Survival Study (4S). AB - OBJECTIVE: Statins have been hypothesized to slow loss of kidney function, however, data is conflicting. This study assessed whether simvastatin reduces rates of kidney function loss in participants with coronary heart disease (CHD). METHODS: We performed a post hoc analysis from the Scandinavian Simvastatin Survival Study, a randomized trial of simvastatin vs. placebo in 4444 men and women with CHD. Kidney disease progression defined as the occurrence of a >or=25% reduction in estimated glomerular filtration rate (GFR) from baseline or development of chronic kidney disease (CKD), defined as estimated GFR <60 mL/min/1.73 m(2). The annual rate of change in estimated GFR was also evaluated. Estimated GFR was determined by the Modified Diet and Renal Disease Study equation. RESULTS: Change in kidney function could be calculated in 3842 subjects, of whom 409 had CKD at baseline. In the whole population, simvastatin significantly reduced the frequency of a >or=25% decline in kidney function [adjusted odds ratio (OR) 0.68, 95% confidence interval (CI), 0.50-0.92; p=0.01] during 5.5 years of follow-up. The adjusted annualized least-square mean decline in estimated GFR in the simvastatin group was lower than that in the placebo group (0.34 mL/min/1.73 m(2)/year vs. 0.41 mL/min/1.73 m(2)/year, respectively; p=0.02). There was no significant effect of simvastatin on kidney function in the subgroup of participants (n=363), who developed incident CKD at the end of follow up [adjusted OR: 0.89 (95% CI 0.70-1.14; p=0.36)]. CONCLUSION: Simvastatin was associated with a moderate protective effect on the rate of kidney function loss in participants with CHD. PMID- 19124122 TI - Alcohol consumption is directly associated with carotid intima-media thickness in Finnish young adults: the Cardiovascular Risk in Young Finns Study. AB - OBJECTIVES: There is substantial epidemiological data suggesting a J- or U-shaped association between alcohol consumption and coronary events. However, some studies in experimental animals suggest that alcohol may increase atherosclerosis. Therefore, our aim was to study whether alcohol consumption is associated with carotid intima-media thickness (IMT), marker of subclinical atherosclerosis, in young, healthy adults. METHODS: Alcohol consumption, carotid IMT and conventional cardiovascular risk factors were investigated in 2074 subjects, aged 24-39 years. RESULTS: In subjects consuming none, >0 to <2, 2 to <4 or >or=4 units of alcohol per day, the respective carotid IMT values were 0.57+/-0.004, 0.59+/-0.003, 0.59+/-0.006, and 0.60+/-0.012 mm (mean+/-S.E.M., P<0.0001 for increasing IMT trend across alcohol consumption categories). This direct association between alcohol consumption and IMT was independent of age, sex and several cardiovascular risk factors, e.g. blood pressure, LDL cholesterol, HDL-cholesterol, BMI, smoking, CRP and insulin (P=0.008 in multivariable regression model). The frequencies of drinking wine or strong alcohol beverages (respective P-values 0.03 and 0.01 for increasing IMT trend across beverage consuming frequency) were directly correlated with carotid IMT in models adjusted for age, sex and risk factors. CONCLUSIONS: We found a direct relationship between alcohol consumption and carotid IMT in young adults. This association was independent of cardiovascular risk factors suggesting that in young healthy adults alcohol consumption may have pro-atherogenic effects. PMID- 19124123 TI - Risk factor correlates of platelet and leukocyte markers assessed by flow cytometry in a population-based sample. AB - BACKGROUND: Platelet and leukocyte products are involved in atherothrombosis. However, the determinants of platelet and leukocyte markers assessed by flow cytometry have not been documented in a population-based sample. METHODS AND RESULTS: We performed flow cytometry on blood from participants (n=1894) in the Atherosclerosis Risk in Communities (ARIC) Carotid MRI Study. Cellular aggregates and multiple platelet and leukocyte markers, such as myeloperoxidase in granulocytes and toll-like receptor-4, CD14, and CD45 in monocytes, were quantified. Their cross-sectional associations with demographic and risk factors were assessed using multiple linear regression. Mean values of most cellular markers and aggregates were considerably higher in blacks than whites (p<0.01). There were some differences in cellular markers between men and women, but little association with age. LDL-cholesterol was associated positively with several markers (toll-like receptor-4 and myeloperoxidase in granulocytes and CD162 in lymphocytes). Cholesterol-lowering therapy tended to show opposite associations. Smokers had much higher granulocyte myeloperoxidase than nonsmokers. However, most other correlations between risk factors and cellular markers were nonsignificant. CONCLUSIONS: Race/ethnicity, sex, and to a lesser degree LDL cholesterol and cholesterol-lowering therapy, but few other risk factors, were correlated with markers of cellular activation in this population-based study. PMID- 19124124 TI - Exogenous in vivo NO-donor treatment preserves p53 levels and protects vascular cells from apoptosis. AB - OBJECTIVE: Nitric oxide (NO) is critical in cardiovascular protection. However, NO production is impaired in atherosclerosis resulting in thrombosis, vasoconstriction, and restenosis. Exogenous NO-donors (NOd) have proven protection against ischemia and thrombosis. However, their effect on vascular cell function remains unknown. Our objective was to determine the effect of NOd on vascular smooth muscle cell (VSMC) survival. METHODS: Pigs (N=16) were randomly distributed in the following treatment groups: (I) LA419 (nitratethiol; 10-day p.o. 0.9 mg/kg bid); (II) LA816 (nitrosothiol; 2h i.v. infusion, 6.6 nmol/(kg min)); (III) nitroglycerine (GTN, 2h i.v. infusion, 2.5mg/kg); and (IV) placebo-control. After sacrifice, pig aortic arch explants were either frozen or incubated with 20% homologous pig serum for different time periods (18h, 2 and 10 days). Bcl2/bax ratio, phosphorylated-p53, cyclin-D2, casp-3, casp-8, and bax mRNA levels and protein expression were evaluated by real-time PCR and western blot, respectively. RESULTS: Within the first 2 days explants from NOd-treated animals had an increased pro-survival Bcl2/bax ratio compared with placebo control (5x higher in LA816 and GTN i.v.-treated animals and 10x higher in 10-day orally treated LA419 animals; P<0.05). Phosphorylated-P53-protein levels were consistently increased by all NOd-treatments vs. placebo-control (P<0.05) and p53 mRNA levels showed an increase in placebo-controls at day 10 while they did not rise in NOd-treated animals (P<0.05). Cyclin-D2, casp-3, and bax mRNA content were 6x lower in NOd-treated animals than in control explants at day-10 (P<0.001). No changes were detected on caspase-8. CONCLUSIONS: Taken together, our findings suggest that exogenous in vivo NO treatment seems to preserve VSMC from mitochondrial-dependent apoptosis and drive cells to quiescence through p53 increase. PMID- 19124125 TI - High apoB/apoA-I ratio is associated with increased progression rate of carotid artery intima-media thickness in clinically healthy 58-year-old men: experiences from very long-term follow-up in the AIR study. AB - OBJECTIVES: The aim of the present study was to investigate the relationship between atherosclerotic progression rate as measured by carotid artery IMT during very long-term follow-up in clinically healthy men and a number of baseline risk factors of potential importance for atherosclerosis progression including apoA-I, apoB, apoB/apoA-I ratio, other lipid variables including LDL particle size, body composition variables, blood pressure, smoking, fasting blood glucose and insulin, and also hsCRP. BACKGROUND: Low-density lipoprotein (LDL) is associated with increased carotid IMT progression rate during long-term follow-up, whereas the relationship between newer biomarkers such as apoB/apoA-I ratio and carotid artery IMT progression rate has been less investigated. METHODS: 58-year-old men identified by screening in the community (n=391) with varying degrees of obesity and insulin sensitivity were examined with high-resolution B-mode ultrasound at baseline and after 3, and 8.9 years of follow-up (n=305 investigated after 8.9 years). The carotid arteries were examined bilaterally, and the mean intima thickness was calculated for 10mm sections of the composite of common carotid arteries and bulbs (IMT(composite)). Serum levels of apoB and apoA-I were measured using a turbidimetric method. Uni- and multi-variable analyses were performed to study the relationship between carotid IMT(composite) progression rate and risk factors. RESULTS: In a multi-variable analysis including all baseline variables only the apoB/apoA-I ratio (p=0.003; beta=0.181, standard error=0.003) and serum insulin (p=0.026; beta=-0.133, standard error=0.000) was significantly related to IMT(composite) progression rate. CONCLUSION: The results indicate that apoB/apoA-I ratio is an important risk factor for predicting atherosclerotic progression rate during very long-term follow-up in clinically healthy middle-aged men. PMID- 19124126 TI - Influence of the distance in a roundhouse kick's execution time and impact force in Taekwondo. AB - Taekwondo, originally a Korean martial art, is well known for its kicks. One of the most frequently used kicks in competition is Bandal Chagui or roundhouse kick. Excellence in Taekwondo relies on the ability to make contact with the opponent's trunk or face with enough force in as little time as possible, while at the same time avoiding being hit. Thus, the distance between contestants is an important variable to be taken into consideration. Thirty-one Taekwondo athletes in two different groups (expert and novice, according to experience in competition) took part in this study. The purpose of this study was to examine both impact force and execution time in a Bandal Chagui or roundhouse kick, and to explore the effect of execution distance in these two variables. A new model was developed in order to measure the force exerted by the body on a load. A force platform and a contact platform were used to measure these variables. The results showed that there are no significant differences in terms of impact force in relation to execution distance in expert competitors. Significant and positive correlations between body mass and impact force (p<.01) seem to mean that novice competitors use their body mass to generate high impact forces. Significant differences were found in competitive experience and execution time for the three different distances of kicking considered in the study. Standing at a certain further distance from the opponent should be an advantage for competitors who are used to kick from a further distance in their training. PMID- 19124127 TI - Modeling of the muscle/tendon excursions and moment arms in the thumb using the commercial software anybody. AB - A biomechanical model of a thumb would be useful for exploring the mechanical loadings in the musculoskeletal system, which cannot be measured in vivo. The purpose of the current study is to develop a practical kinematic thumb model using the commercial software Anybody (Anybody Technology, Aalborg, Denmark), which includes real CT-scans of the bony sections and realistic tendon/muscle attachments on the bones. The thumb model consists of a trapezium, a metacarpal bone, a proximal and a distal phalanx. These four bony sections are linked via three joints, i.e., IP (interphalangeal), MP (metacarpophalangeal) and CMC (carpometacarpal) joints. Nine muscles were included in the proposed model. The theoretically calculated moment arms of the tendons are compared with the corresponding experimental data by Smutz et al. [1998. Mechanical advantage of the thumb muscles. J. Biomech. 31(6), 565-570]. The predicted muscle moment arms of the majority of the muscle/tendon units agree well with the experimental data in the entire range of motion. Close to the end of the motion range, the predicted moment arms of several muscles (i.e., ADPt and ADPo (transverse and oblique heads of the adductor pollicis, respectively) muscles for CMC abduction/adduction and ADPt and FPB (flexor pollicis brevis) muscle for MP extension/flexion) deviate from the experimental data. The predicted moment potentials for all muscles are consistent with the experimental data. The findings thus suggest that, in a biomechanical model of the thumb, the mechanical functions of muscle-tendon units with small physiological cross-sectional areas (PCSAs) can be well represented using single strings, while those with large PCSAs (flat-wide attachments, e.g., ADPt and ADPo) can be represented by the averaged excursions of two strings. Our results show that the tendons with large PCSAs can be well represented biomechanically using the proposed approach in the major range of motion. PMID- 19124128 TI - A novel application of velocity-based force control for use in robotic biomechanical testing. AB - This paper presents a novel application of a velocity-based force control routine used for robotic biomechanical testing. The routine employs a jog function, available from the robot's motion commands, that permits easy adjustment of velocity on each axis. Force and moment targets are achieved by adjusting jog velocities in proportion to force or moment errors while limiting the maximum velocity of the system. The force control jog routine does not require specimen stiffness values and is inherently stable. The performance of the method was shown to be suitable for unconstrained in vitro spine testing in a rabbit model where extremely small motions are necessary to maintain the target force values. The jogging feature on which this work is based is a feature available on most robots and is equally applicable to a serial robot. The simplicity, stability, and performance of this method warrant its consideration for other robotic biomechanical testing applications where force control is required. PMID- 19124129 TI - Use of a programmed temperature vaporizer and an in situ derivatization reaction to improve sensitivity in headspace-gas chromatography. Application to the analysis of chlorophenols in water. AB - In the present work we propose the combined use of a derivatization reaction within the vial of a headspace sampler with a programmed temperature vaporizer (PTV) inlet in the solvent vent mode as a new methodology for obtaining an increase in sensitivity in headspace-gas chromatography (HS-GC) for the analysis of sparingly volatile compounds. As test analytes the following chlorophenols were used: 2-chlorophenol (2CP), 2,4-dichlorophenol (24DCP), 4-chloro-3 methylphenol (4C3MP) and 2,4,6-trichlorophenol (246TCP). The derivatization reaction was carried out with acetic anhydride because it can be carried out in situ in aqueous medium. In the programmed temperature vaporizer inlet, three different liners, one of them empty and the others with materials of different trapping strengths (glass wool and Tenax-TA), were compared. The best results were obtained when an empty liner was used, with better repeatability and S/N ratios. In the case of the liner filled with Tenax-TA, a considerable lack of repeatability was observed, this being attributed to interactions between the derivatized compounds and the adsorbent. The proposed methodology affords very low limits of detection, in the range of a few ng/L for all the compounds, with good precision and accuracy values. PMID- 19124130 TI - Novel beta-cyclodextrin derivative functionalized polymethacrylate-based monolithic columns for enantioselective separation of ibuprofen and naproxen enantiomers in capillary electrochromatography. AB - A novel enantioselective polymethacrylate-based monolithic column for capillary electrochromatography was prepared by ring-opening reaction of epoxy groups from poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolith with a novel beta cyclodextrin derivative bearing 4-dimethylamino-1,8-naphthalimide functionalities. Conditions for the ring-opening reaction with respect to different reaction parameters were thoroughly optimized to obtain high electroosmotic flow, separation efficiency and enantioselectivity for the analytes. The nonaqueous mobile phase composition regarding acetonitrile-methanol ratio and the concentration of electrolyte were examined to manipulate the hydrophobic inclusion and anion-exchange interaction between the analytes and chiral stationary phase. It was observed that in addition to beta-cyclodextrin cavity, the electrostatic interaction exhibited pronounced influence on the enantioseparation of acidic analytes. Acidic enantiomers (ibuprofen and naproxen) could be separated with separation factor (alpha) values up to 1.08 and a maximum separation efficiency of 86000 plates/m could be achieved. PMID- 19124131 TI - The 3-amino-derivative of gamma-cyclodextrin as chiral selector of Dns-amino acids in electrokinetic chromatography. AB - The enantioseparation of the enantiomeric pairs of 10 Dns derivatives of alpha amino acids was successfully carried out by using for the first time the 3-amino derivative of the gamma-cyclodextrin. The effects of pH and selector concentration on the migration times and the resolutions of analytes were studied in detail. 3-Deoxy-3-amino-2(S),3(R)-gamma-cyclodextrin (GCD3AM) shows very good chiral recognition ability even at very low concentrations at all the three investigated values of pH, as shown by the very large values of selectivity and resolution towards several pairs of amino acids. The role played by the cavity, the substitution site and the protonation equilibria on the observed properties of chiral selectivity, on varying the specific amino acid involved, is discussed. PMID- 19124132 TI - Open source simulation tool for electrophoretic stacking, focusing, and separation. AB - We present the development, formulation, and performance of a new simulation tool for electrophoretic preconcentration and separation processes such as capillary electrophoresis, isotachophoresis, and field amplified sample stacking. The code solves the one-dimensional transient advection-diffusion equations for multiple multivalent weak electrolytes (including ampholytes) and includes a model for pressure-driven flow and Taylor-Aris dispersion. The code uses a new approach for the discretization of the equations, consisting of a high resolution compact scheme which is combined with an adaptive grid algorithm. We show that this combination allows for accurate resolution of sharp concentration gradients at high electric fields, while at the same time significantly reducing the computational time. We demonstrate smooth, stable, and accurate solutions at current densities as high as 5000A/m(2) using only 300 grid points, and a 75-fold reduction in computational time compared with equivalent uniform grid techniques. The code is available as an open source for free at http://microfluidics.stanford.edu. PMID- 19124133 TI - Fullerenes for aromatic and non-aromatic N-nitrosamines discrimination. AB - The detection of N-nitrosamines (NAms) in water supplies is an environmental and public health issue because many NAms are classified as probable human carcinogens. Non-aromatic (aliphatic and cyclic) NAms are more toxic than aromatic ones as their maximum admissible concentration is limited in drinking water (20-2000ngL(-1)). From that premise, a simple and novel method to discriminate between both fractions of NAms according to their toxicity was proposed. An automatic solid-phase extraction unit containing two sequential sorbent columns was constructed. A sample volume of 25mL was passed through a C(60) fullerene column in which only the aromatic fraction was retained, and the effluent was then passed through a Merck LiChrolut EN column where the non aromatic fraction was retained. Following elution of the non-aromatic NAms with 150microL of ethyl acetate-acetonitrile (9:1), 1microL of the extract was injected into a GC/MS. A comparative study of C(60) and C(70) fullerenes and nanotubes revealed C(60) fullerene to be the best choice to selectively retain the aromatic fraction. The method exhibits a linear range of 15-20,000ngL(-1); limits of detection of 4-15ngL(-1); and an RSD of approximately 5%. Recoveries throughout the whole method were between 95% and 102% for six non-aromatic NAms spiked into several types of waters. Our study demonstrates that a simple and fast SPE system (10min per sample) with a customary GC-MS instrument permits the quantification of these amines in complex matrices with considerable sensitivity and selectivity. PMID- 19124134 TI - Detection of porcine circovirus 2 in mammary and other tissues from experimentally infected sows. AB - The aim of this study was to determine whether porcine circovirus 2 (PCV2) may infect the mammary gland of sows and be shed in the milk. Six pregnant sows were inoculated intranasally with PCV2 three weeks before their expected farrowing date and two further sows acted as uninfected controls. The animals remained clinically healthy and farrowed normally. Milk samples were collected from all sows on the first, second and third days of lactation. PCV2 DNA was detected in the milk of infected sows from day 1 of lactation but not in the milk of uninfected controls. PCV2 antigen and DNA were detected in the mammary gland and other tissues by immunohistochemistry and in-situ hybridization, respectively. Simultaneous detection of viral protein and DNA provided molecular evidence of PCV2 infection and replication within these tissues. PMID- 19124135 TI - Influence of intensity on children's sensitivity to happy, sad, and fearful facial expressions. AB - Most previous studies investigating children's ability to recognize facial expressions used only intense exemplars. Here we compared the sensitivity of 5-, 7-, and 10-year-olds with that of adults (n=24 per age group) for less intense expressions of happiness, sadness, and fear. The developmental patterns differed across expressions. For happiness, by 5 years of age, children were as sensitive as adults even to low intensities. For sadness, by 5 years of age, children were as accurate as adults in judging that the face was expressive (i.e., not neutral), but even at 10 years of age, children were more likely to misjudge it as fearful. For fear, children's thresholds were not adult-like until 10 years of age, and children often confused it with sadness at 5 years of age. For all expressions, including even happy expressions, 5- and 7-year-olds were less accurate than adults in judging which of two expressions was more intense. Together, the results indicate that there is slow development of accurate decoding of subtle facial expressions. PMID- 19124136 TI - Floating marine debris in fjords, gulfs and channels of southern Chile. AB - Floating marine debris (FMD) is reported from all oceans. The bulk of FMD are plastics, which due to their longevity cause multiple negative impacts on wildlife and environment. Identifying the origins of FMD (land- or sea-based) is important to take the necessary steps to diminish their abundance. Using ship surveys we examined the abundance, composition and distribution of FMD during the years 2002-2005 in the fjords, gulfs and channels of southern Chile. Abundances of FMD were relatively high compared with other studies, ranging from 1 to 250 items km(-2). The majority (approximately 80%) of FMD was composed of styrofoam (expanded polystyrene), plastic bags and plastic fragments. Styrofoam, which is intensively used as flotation device by mussel farms, was very abundant in the northern region but rarely occurred in the southern region of the study area. Food sacks from salmon farms were also most common in the northern region, where approximately 85% of the total Chilean mussel and salmon harvest is produced. Plastic bags, which could be from land- or sea-based sources, were found throughout the entire study area. Our results indicate that sea-based activities (mussel farming and salmon aquaculture) are responsible for most FMD in the study area. In order to reduce FMDs in the environment, in addition to stronger legislation and identification of potential sources, we suggest environmental education programs and we encourage public participation (e.g. in beach surveys and clean-ups). PMID- 19124137 TI - Occurrence and concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in coastal marine sediments in Kuwait. PMID- 19124139 TI - Cerebral immunoglobulin light chain amyloid angiopathy-related hemorrhages. AB - INTRODUCTION: Cerebral amyloid angiopathy (CAA) is a common cause of intracerebral hemorrhage (ICH) particularly in elderly patients. In CAA-related hemorrhages, amyloid deposits in the brain vessel walls mainly contain amyloid beta-protein (A-beta). Rarely other forms of amyloid substances have been reported in sporadic CAA-related hemorrhages. METHODS: We report the case of a 44 year-old patient with recurrent ICH who had surgical evacuation of a large frontal hematoma. Following surgery, samples from the hematoma and adjacent cerebral cortex were obtained for histopathological examination. RESULTS: Within the recent hemorrhage, a few arteriolar walls were thickened with an amyloid deposit that was immunostained for immunoglobulin (Ig) M and light chain lambda. In the wall of some vessels, around the amyloid deposits, as well as in the adjacent cerebral cortex, there was an infiltration by monotypic lymphocytes and plasma cells expressing IgM and light chain lambda. No amyloid deposition was found outside the hemorrhage. There was no evidence of multiple myeloma, B-cell malignancy, or systemic amyloidosis. CONCLUSIONS: Recurrent ICH may be due to amyloid deposition of IgM lambda produced by monotypic proliferation of lymphocytes and plasma cells purely localized to the brain. PMID- 19124140 TI - [Homonymous hemianopia and posterior cortical atrophy]. AB - INTRODUCTION: Posterior cortical atrophy (PCA) is a clinically and radiologically defined syndrome, in which predominant symptoms focus on higher visual dysfunction with progressive course and association with cortical atrophy or hypometabolism that predominates in the posterior part of the hemispheres. Homonymous hemianopia (HH) has rarely been described in this syndrome. METHODS: We report on six patients (four females, two males, aged 63 to 80) referred for visual disorder which led to demonstration of HH using perimetry testing. These patients were followed for 1 to 5 years after discovery of HH. Brain imaging with MRI or CT scan was obtained in the six cases and a SPECT scan was performed in four cases. RESULTS: HH was left-sided in four cases and right-sided in two cases. Associated symptoms related to higher visual dysfunction were simultagnosia, alone or as part of a full Balint's syndrome, alexia, constructional apraxia, dressing apraxia, visual form agnosia, prosopagnosia and hemispatial neglect. These symptoms were mild at onset but invariably worsened with disease progression. Dementia eventually developed in all cases. The clinical diagnosis was probable Alzheimer's disease in five cases and corticobasal degeneration in one case. Radiology showed posterior cortex atrophy in all cases as well as reduced cerebral blood flow in the same region, with an asymmetrical pattern compatible with the side of HH. CONCLUSION: Elementary cortical lesions in PCA can develop mainly in the associative visual areas and even in the primary visual area, resulting in HH. HH has rarely been documented in PCA, but its prevalence would probably be higher if systematic search was conducted. Apparently isolated HH of insidious onset should suggest PCA and lead to neuropsychological testing and search for discrete atrophic changes of the posterior cortex on MRI as well as for metabolic alterations with SPECT or PET. PMID- 19124141 TI - [Susac syndrome: study of five cases]. AB - INTRODUCTION: Susac syndrome is a rare microangiopathy, responsible for small cerebral, retinal and cochlear infarcts. The classic clinical triad includes multiple neurologic signs (from headaches to coma), retinal branch occlusions and sensorineural hearing loss. METHODS: We report a series of five patients with Susac syndrome followed in our department from 1997 to 2007. RESULTS: There were four women and one man (mean age at onset: 35.2 years). Clinical symptoms at onset were neurological (n=1), ophthalmological (n=1), auditory (n=1) and clinical triad (n=2). Neurologic symptoms included encephalopathy (n=2), headache (n=5), transient ischemic attacks (n=1). Brain MRI showed T2 lesions in the white and grey matter, corpus callosum and gadolinium-enhanced punctiform lesions. Cerebrospinal fluid contained an elevated protein level in three cases. Immunologic treatments (steroids [n=4], cylophosphamid [n=3], intravenous immunoglobulins [n=5]) associated with aspirin and/or oral anticoagulants, despite early relapses (n=2), led to dramatic clinical improvement (n=5). CONCLUSION: Due to its polymorphism the SS is difficult to diagnose when the clinical triad is lacking. In the absence of clinical trial and consensus treatment is empiric and based on supposed pathogenesis. PMID- 19124142 TI - [Intravenous thrombolysis with rt-PA in stroke: experience of the Nice stroke unit]. AB - INTRODUCTION: Intravenous thrombolysis with rt-Pa in stroke has been approved in France since 2002. We report an evaluation of our practice. We have tried to identify predictive factors of dependence and death, and to compare our results with the data of the literature. PATIENTS AND METHOD: All patients treated with intravenous rt-PA within the first 270min after the stroke onset were included. Univariate, then multivariate analyses were performed to determine the variables influencing the functional outcome at 3 months follow-up, according to a dichotomy established from the modified Rankin scale. RESULTS: One hundred and forty-two patients were included in this study (mean initial National Institute of Health Stroke Scale [NIHSS]: 15). Fifty percent had a Rankin score higher than 2 at 3 months follow-up. NIHSS above 12, glycemia of at least 120mg/l, and systolic blood pressure above 160mmHg at admission were identified as independent predictive factors of poor functional outcome. Less than 4 points decrease of NIHSS proved to be a simple and early predictor of poor functional outcome at 3 months follow-up. CONCLUSIONS: In terms of safety and efficacy the data issuing from the daily activity of our stroke unit are comparable with those of clinical trials. PMID- 19124143 TI - Determination of flame-retardant hexabromocyclododecane diastereomers in textiles. AB - To establish a concise and rapid procedure to analyze hexabromocyclododecane (HBCD) diastereomers in flame-retarded textiles, three different methods of extraction-Soxhlet, ultrasonic, and soaking extractions with toluene and dichloromethane (DCM)-were compared. During Soxhlet extraction using toluene, the percent contribution of alpha-HBCD to total HBCDs increased slightly and that of gamma-HBCD decreased, indicating that gamma-HBCD was isomerized to some extent at the boiling point of toluene (110.6 degrees C). For ultrasonic extraction, the temperature of the water bath can easily increase over time during the procedure, which might lead to undesirable effects. Therefore, we considered soaking extraction with DCM to be the most facile procedure to analyze HBCD diastereomers in textiles. Using the method established in this study, commercially available textiles in Japan (n=10) were analyzed to understand the actual composition of HBCD contents and its diastereomer profiles. With the exception of one textile sample, HBCDs were detected in all the samples analyzed, with concentrations ranging from 22000 to 43000 mg kg(-1) (i.e. 2.2-4.3%). We found a higher proportion of the alpha-diastereomer in most textile products compared with that of commercial HBCD mixtures, indicating that gamma-HBCD isomerized to alpha diastereomer by heating processes to incorporate the commercial formulation into treated materials or that the alpha-diastereomer preferentially absorbed onto textile materials during the manufacturing of flame-retarded consumer products. PMID- 19124144 TI - Effects of microcystin-LR on the expression of P-glycoprotein in Jenynsia multidentata. AB - The multixenobiotic resistance phenomenon (MXR) related to the P-glycoprotein multidrug transporter protein (P-gp) has been identified and characterized in several aquatic organisms. In the present work, we prove the presence of a P-gp in liver, gills and brain of Jenynsia multidentata by Western Blot and RT-PCR. A 170 kDa protein has been found in liver and gills while in brain a approximately 80 kDa protein has been detected. The partial nucleotide sequence obtained in this autochthonous fish showed high similarity ranging from 83% to 92% with other fishes. In addition, P-gp expression in this fish was evaluated after time and dose-dependent exposures to the cyanotoxin microcystin-LR. Individuals were exposed to MC-LR at concentrations of 2, 5 and 10 microg L(-1) for 24h and for 6, 12 and 24h at 2 microg L(-1) MC-LR. Changes in P-gp expression were observed in liver, gills and brain. However, this response was tissue specific. Only in gills of J. multidentata P-gp expression, measured either by real-time RT-PCR or Western Blot, was significantly higher compared to controls at most tested times and doses. A 3-fold increase with respect to controls was found at 12h by RT-PCR and after 24h by Western Blot. In dose-dependent experiments the maximum P-gp expression was observed at 2 microg L(-1) MC-LR, measured by both RT-PCR and Western Blot. In the liver, P-gp protein levels were significantly increased after 24h of exposure, at every toxin dose tested. Thus, probably longer exposures would show also significant increases in this tissue. Considering these results we can propose that P-gp belongs to the defence system involved in the response to MC-LR in J. multidentata. PMID- 19124145 TI - Possible role of denitrification on aerobic granular sludge formation in sequencing batch reactor. AB - The aim of this work was to evaluate and quantify the influence of denitrification of nitrate on the aerobic granular sludge development. Two parallel sequencing batch reactors (SBRs) were fed with synthetic wastewater, the first reactor (R1) receiving nitrate at a fixed concentration whereas the second reactor (R2) did not receive any external source of nitrate. Both systems were working at the same operational conditions with a low specific air velocity (0.07cms(-1)), a relatively low and acceptable oxygen concentration (1.8+/ 0.8mgL(-1)) and without significant biomass selection pressure (minimal velocity 0.4mh(-1)). The results showed that the sludge settling properties of R1 were comparable to granular sludge (settling velocity approximately 10.6mh(-1), SVI(5) approximately 13-30mLg(-1), SVI(30) approximately 11-28mLg(-1) and SVI(5)/SVI(30) ratio approximately 1.05-1.1). In the opposition, biomass of the reactor R2 reveals more traditional properties. Nitrogen measurements revealed that a significant denitrification (up to 80% of fed nitrate) occurred in R1 in the core of aggregates, as aerobic condition was maintained in the bulk. After the mean particle size was reduced from 250 to 125microm by an increase of mechanical stirring rate, it was shown that denitrification decreased from 80% to 20%. These parallel experiments definitively proved that the presence of nitrate in SBRs can assist the densification of the biological aggregates in aerobic condition. The presence of nitrate (commonly provided by nitrification) should be considered as a possible factor which helps to maintain the granulation process. PMID- 19124146 TI - Structural characterization and thermal stabilities of the isomers of the brominated flame retardant 1,2,5,6-tetrabromocyclooctane (TBCO). AB - 1,2,5,6-Tetrabromocyclooctane (TBCO) is a commercial brominated flame retardant that is employed mainly as an additive in textiles, paints and plastics. Very little is known about its presence or behavior in the environment or its analysis. TBCO can exist as two diastereomers, the stereochemistries of which have not been previously reported. We have named the first eluting isomer, under HPLC conditions, as alpha-TBCO (alpha-TBCO) and the later eluting isomer as beta TBCO (beta-TBCO) when using an Acquity UPLC BEH C(18) column with methanol/acetonitrile/water as the mobile phase. The structural elucidation of these two isomers was accomplished by 1H NMR spectroscopy, GC/MS, LC/MS and X-ray structure determinations. alpha-TBCO is (1R,2R,5S,6S)-1,2,5,6 tetrabromocyclooctane and beta-TBCO is rac-(1R,2R,5R,6R)-1,2,5,6 tetrabromocyclooctane. As with some other brominated cycloaliphatic compounds, TBCO is thermally labile and the isomers easily interconvert. A thermal equilibrium mixture of alpha- and beta-TBCO consists of approximately 15% and 85% of these isomers, respectively. Separation of the two diastereomers, with minimal thermal interconversion between them, is achievable by careful selection of GC capillary column length and injector temperature. LC/MS analyses of TBCO also presents an analytical challenge due to poor resolution of the isomers on chromatographic stationary phases, and weak intensity of molecular ions (or major fragment ions) when using LC-ESI/MS. Only bromide ions were seen in the mass spectra. APCI and APPI also failed to produce the molecular ion with sufficient intensity for identification. PMID- 19124147 TI - Environmental fate of the antiviral drug Tamiflu in two aquatic ecosystems. AB - The antiviral drug Tamiflu (Oseltamivir Phosphate, OP), has been indicated by the World Health Organization as a first-line defense in case of an avian influenza pandemic. Recent studies have demonstrated that Oseltamivir Carboxylate (OC), the active metabolite of the prodrug OP, has the potential to be released into water bodies. The present laboratory study focused on basic processes governing the environmental fate of OC in surface water from two contrasting aquatic ecosystems of northern Italy, the River Po and the Venice lagoon. Results of this study confirmed the potential of OC to persist in surface water. However, addition of 5% of sediments resulted in rapid OC degradation. Estimated half-life of OC in water/sediment of the River Po was 15 days. After three weeks of incubation at 20 degrees C, more than 8% of (14)C-OC evolved as (14)CO(2) from water/sediment samples of the River Po and Venice lagoon. At the end of the 21-day incubation period, more than 65% of the (14)C-residues were recovered from the liquid phase of both Po and Venice water/sediment samples. OC was moderately retained onto coarse sediments from the two sites. In water/sediment samples of the River Po and Venice lagoon treated with (14)C-OC, more than 30% of the (14)C-residues remained water-extractable after three weeks of incubation. The low affinity of OC to sediments suggests that presence of sediments would not reduce its bioavailability to microbial degradation. PMID- 19124149 TI - Autonomic innervation of the fish gut. AB - The enteric nervous system follows a similar overall arrangement in all vertebrate groups. In fish, the majority of nerve cell bodies are found in the myenteric plexus, innervating muscles, blood vessels and glands. In this review, I describe similarities and differences in size, shape and transmitter content in enteric neurons in different fish species and also in comparison with other vertebrates, foremost mammals. The use of different histological and immunochemical methods is reviewed in a historical perspective including advantages and disadvantages of different methods. Lately, zebrafish have become an important model species for developmental studies of the nervous system, including the enteric nervous system, and this is briefly discussed. Finally, examples of how the enteric nervous system controls gut activity in fish is presented, focussing on the effect on gastrointestinal motility. PMID- 19124148 TI - Interaction of tomato lectin with ABC transporter in cancer cells: glycosylation confers functional conformation of P-gp. AB - Phospho-glycoprotein (P-gp) is a polytopic plasma membrane protein whose overexpression causes multidrug resistance (MDR) responsible for the failure of cancer chemotherapy. P-gp 170 is a member of the ATP-binding cassette (ABC) transporter superfamily and has two potentially interesting regions for drugs interfering with its efflux function, namely the oligosaccharides on the first extracellular loop with unknown function and the two intracellular ATP-binding regions providing the energy for drug efflux function. The polylactoseamine oligosaccharides on the first loop can specifically bind the tomato lectin (TL). The P-gp efflux activities of TL-pre-treated MDR resistant cells were measured in the presence of structurally unrelated resistance modifiers such as phenothiazines, terpenoids and carotenoids. The inhibition of efflux activity was measured via the increased rhodamine uptake by mouse lymphoma cells transfected in human MDR1 gene and in human brain capillary endothelial cells. The tested resistance modifiers inhibit the function of ABC transporter resulting in increased R123 accumulation in MDR1 expressing cells. TL prevented the inhibitory action of phenothiazine and verapamil on brain capillary endothelial and MDR1 lymphoma cells, presumably due to the stabilization of the functional active conformation of P-gp. Our results indicate that the polylactosamine chains of P gp are part of the functionally active protein conformation. PMID- 19124150 TI - Spatial and cure-time distribution of dynamic-mechanical properties of a dimethacrylate nano-composite. AB - OBJECTIVE: The purpose of this study was to evaluate a nano-filled dental composite, with varying cure irradiation-time, in terms of the spatial distribution of dynamic-mechanical properties determined at nanometre scale and the resultant distinction between filler, matrix and inter-phase regions. MATERIALS AND METHODS: Specimen groups (n=5) of the composite Filtek Supreme XT were cured in 2mm deep molds for 5, 10, 20 and 40s, and stored for 24h in distilled water at 37 degrees C. Properties were measured at 2mm depth, on the lower specimen surfaces. Nano-dynamic-mechanical parameters (complex, storage and loss modulus, tandelta) were determined at an array of 65,000 locations in a 5microm x 5microm area. Micro-mechanical properties (hardness, modulus of elasticity, creep and elastic/plastic deformation) were also measured and additionally the real-time degree of cure, by ATR-FTIR, for 10min after photo initiation and after storage. RESULTS: The spatial distribution of nano-dynamic mechanical properties varied significantly enabling four distinguishable matrix, filler-cluster and inter-phase regions to be identified. Proceeding from matrix to filler-cluster locations, complex-moduli increased linearly and loss-factors decreased linearly, consistent with visco-elastic composite theory. Curing time strongly affected all measured properties at 2mm depth. The organic matrix was shown to be inhomogeneous for all curing times. By increasing cure-time, the proportion of less well polymerized area decreased from 37.7 to 1.1%, resulting in a more homogeneous organic matrix. SIGNIFICANCE: The experimentally observed graduated transition, in complex modulus and related dynamic-mechanical properties, across the matrix - inter-phases - filler-cluster regions is conducive to low internal stresses, in contrast to the abrupt modulus transitions anticipated or observed in many other particulate composite structures. The identification of these phase-regions provides a realistic basis for accurate nano- and micro-mechanical computational modelling. PMID- 19124151 TI - Mixed peptide-chitosan membranes to mimic the biological activities of a multifunctional laminin alpha1 chain LG4 module. AB - Laminin alpha1 chain LG4 module is multifunctional and interacts with syndecans and integrin alpha2beta1 via AG73 (RKRLQVQLSIRT) and EF-I (DYATLQLQEGRLHFMFDLG) sites, respectively. Here, we conjugated the AG73 and EF1zz (ATLQLQEGRLHFXFDLGKGR, X: Nle) peptides on a chitosan membrane in various ratios to develop an LG4 mimic biomaterial. The AG73-chitosan membrane promoted strong cell attachment with membrane ruffling and the EF1zz-chitosan membrane promoted integrin-mediated cell adhesion with well-organized actin stress fibers. When AG73 and EF1zz were conjugated on a chitosan membrane with 1:9 molar ratio, the mixed peptide-chitosan membrane promoted the strong cell attachment and neurite outgrowth similar to that on the recombinant LG4 protein. Well-organized actin stress fibers and vinculin accumulated focal contacts were observed in the cells attached on the AG73:EF1zz (molar ratio=1:9)-chitosan membrane. These results suggest that the mixed peptide-chitosan membrane interacts with both syndecans and integrin alpha2beta1 and mimics the cell adhesion of a multifunctional LG4 protein. The mixed peptide-chitosan approach has potential as a multifunctional biomaterial for cell and tissue engineering. PMID- 19124152 TI - Bone healing induced by local delivery of an engineered parathyroid hormone prodrug. AB - Regenerative medicine requires innovative therapeutic designs to accommodate high morphogen concentrations in local depots, provide their sustained presence, and enhance cellular invasion and directed differentiation. Here we present an example for inducing local bone regeneration with a matrix-bound engineered active fragment of human parathyroid hormone (PTH(1-34)), linked to a transglutaminase substrate for binding to fibrin as a delivery and cell-invasion matrix with an intervening plasmin-sensitive link (TGplPTH(1-34)). The precursor form displays very little activity and signaling to osteoblasts, whereas the plasmin cleavage product, as it would be induced under the enzymatic influence of cells remodeling the matrix, was highly active. In vivo animal bone-defect experiments showed dose-dependent bone formation using the PTH-fibrin matrix, with evidence of both osteoconductive and osteoinductive bone-healing mechanisms. Results showed that this PTH-derivatized matrix may have potential utility in humans as a replacement for bone grafts or to repair bone defects. PMID- 19124153 TI - Preferential cell attachment to nitrogen-doped diamond-like carbon (DLC:N) for the measurement of quantal exocytosis. AB - Electrochemical measurement of transmitter or hormone release from individual cells on microchips has applications both in basic science and drug screening. High-resolution measurement of quantal exocytosis requires the working electrode to be small (cell-sized) and located in immediate proximity to the cell. We examined the ability of candidate electrode materials to promote the attachment of two hormone-secreting cell types as a mechanism for targeting cells for to recording electrodes with high precision. We found that nitrogen-doped diamond like carbon (DLC:N) promoted cell attachment relative to other materials tested in the rank order of DLC:N>In(2)O(3)/SnO(2) (ITO), Pt>Au. In addition, we found that treating candidate electrode materials with polylysine did not increase attachment of chromaffin cells to DLC:N, but promoted cell attachment to the other tested materials. We found that hormone-secreting cells did not attach readily to Teflon AF as a potential insulating material, and demonstrated that patterning of Teflon AF leads to selective cell targeting to DLC:N "docking sites". These results will guide the design of the next generation of biochips for automated and high-throughput measurement of quantal exocytosis. PMID- 19124154 TI - A DNA nanomachine induced by single-walled carbon nanotubes on gold surface. AB - Single-walled carbon nanotubes (SWNTs) can selectively induce human telomeric i motif DNA formation at pH 7.0. Based on this property, we design a DNA nanomachine induced by SWNTs on gold surface. The motor DNA is human telomeric G quadruplex DNA. The reversible hybridization between the motor DNA and its complementary human telomeric i-motif DNA can be modulated by SWNTs without changing solution pH. Up to now, to our knowledge, there is no report to show that a DNA nanomachine is induced by SWNTs or a DNA nanomachine can detect i motif formation at pH 7.0. Our work may provide a new concept for designing an SWNT-induced DNA nanomachine and for the detection of i-motif DNA structure at pH 7.0. DNA hybridization, conformational transition and i-motif formation have been characterized on surface or in solution by fluorescence confocal microscopy, circular dichroism, DNA melting and gel electrophoresis. The folding and unfolding kinetics of the DNA nanomachine on gold surface were studied by Fourier transform-surface plasmon resonance (FT-SPR). All these results indicate that SWNTs can induce the DNA nanomachine to work efficiently and reversibly. Therefore our work will provide new insights into the design and application of SWNT-induced DNA nanodevice under physiological conditions. PMID- 19124155 TI - Tubular micro-scale multiwalled carbon nanotube-based scaffolds for tissue engineering. AB - In this study we have prepared a tubular knitted scaffold from a 9 ply multiwalled carbon nanotube (MWCNT) yarn and a composite scaffold, formed by electrospinning poly(lactic-co-glycolic acid) (PLGA) nanofibres onto the knitted scaffold. Both structures were assessed for in vitro biocompatibility with NR6 mouse fibroblast cells for up to 22 days and their suitability as tissue engineering scaffolds considered. The MWCNT yarn was found to support cell growth throughout the culture period, with fibroblasts attaching to, and proliferating on, the yarn surface. The knitted tubular scaffold contained large pores that inhibited cell spanning, leading to the formation of cell clusters on the yarn, and an uneven cell distribution on the scaffold surface. The smaller pores, created through electrospinning, were found to promote cell spanning, leading to a uniform distribution of cells on the composite scaffold surface. Evaluation of the electrical and mechanical properties of the knitted scaffold determined resistance levels of 0.9 kOmega/cm, with a breaking load and extension to break approaching 0.7N and 8%, respectively. The PLGA/MWCNT composite scaffold presented in this work not only supports cell growth, but also has the potential to utilize the full range of electrical and mechanical properties that carbon nanotubes have to offer. PMID- 19124156 TI - Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression. AB - Expression of the anti-inflammatory cytokine IL-10 is suppressed by the pro inflammatory interferon gamma but the mechanism of this action is unknown. We analysed activity of IL-10 promoter luciferase reporter constructs in transfected RPMI 8226.1 B cells that were treated at -2 h with IFNgamma (1000 U/ml) followed by stimulation with LPS (100 ng/ml) at 0 h. IFNgamma treatment suppressed LPS induced IL-10 promoter activity in a construct carrying the -1044 promoter and also one containing the -195 promoter. The suppression was independent of the IRF motif at -182 but involved the Stat-motif at -120. In gelshift analysis this Stat motif did bind LPS-induced Stat3 and with IFNgamma treatment it did, in addition, bind Stat1. ChIP analysis for detection of transcription factor binding to chromatin in intact cells demonstrated Stat3 binding to the proximal IL-10 promoter when cells are stimulated with LPS only. Treatment with IFNgamma only led to Stat1 binding in ChIP analysis and treatment with IFNgamma plus LPS led to reduced Stat3 binding while Stat1 binding remained high. Finally, LPS-induced activity of the trimeric Stat-motif in front of the luciferase reporter was suppressed by IFNgamma. These data demonstrate that IFNgamma down-regulates expression of the IL-10 gene by a novel mechanism that involves displacement of transactiving Stat3 by IFNgamma-induced Stat1. PMID- 19124157 TI - Porcine Fc gammaRIII isoforms are generated by alternative splicing. AB - The Fc gammaRIII plays an essential role in antibody-mediating effector functions of immune cells. Here, we report that transcripts encoding porcine Fc gammaRIII isoforms are generated by alternative splicing. Fc gammaRIII a.1 is expressed on the cell surface while Fc gammaRIII a.2 is secreted from the transfected cells due to a partial deletion of the transmembrane domain. Interestingly, a putative soluble Fc gammaRIII (sCD16) is detected in circulating plasma. Both Fc gammaRIII a.2 and sCD16 exhibit a similar molecular mass (approximately 35 kDa) and contain the extracellular D2 domains that are structurally intact. Despite the D2 domain deletion, Fc gammaRIII a.3 associates with Fc gammaRIII a.1. Hence, these results suggest one possibility that three Fc gammaRIII isoforms differentially modulate Fc gammaRIII-mediated immune responses in the porcine system. Furthermore, we demonstrate that a cytolytic triggering G7 monoclonal antibody recognizes the D2 domain that is responsible for interacting with the immune complexes and subsequent activations of porcine innate immune cells. This result suggests that the D2 domain is a target region to develop therapeutic antibodies that regulate the FcR-mediated immune responses. PMID- 19124158 TI - Effects of the loss of Atox1 on the cellular pharmacology of cisplatin. AB - Previous work has demonstrated that the copper (Cu) transporters Ctr1, Atp7a and Atp7b regulate the cellular pharmacology of cisplatin (CDDP) by mediating its uptake and efflux. It was also shown that, in the process of uptake by Ctr1, CDDP triggers the rapid proteasomal degradation of its own transporter. The current study examined the role of the metallochaperone Atox1 in the regulation of uptake, efflux and subcellular distribution of CDDP by using a pair of fibroblast cell lines established from Atox1(+/+) and Atox1(-/-) mice. Atox1 is a metallochaperone that is known to play a central role in distributing Cu within the cells and was recently shown to act as a Cu-dependent transcription factor. Loss of Atox1 increased Cu accumulation and reduced efflux. In contrast, loss of Atox1 reduced the influx of CDDP and subsequent accumulation in vesicular compartments and in DNA. Loss of Atox1 was found to block the CDDP-induced down regulation of Ctr1. Ctr1 was found to be polyubiquitinated in an Atox1-dependent manner during CDDP exposure. In conclusion, Atox1 is required for the polyubiquitination of Ctr1 and the Ctr1-mediated uptake of CDDP. PMID- 19124159 TI - Expression of IFN-gamma, TNF-alpha, IL-10 and TGF-beta in lymph nodes associates with parasite load and clinical form of disease in dogs naturally infected with Leishmania (Leishmania) chagasi. AB - American visceral leishmaniasis is a zoonosis of the New World. Dogs are the main reservoir of the disease and there is much interest in the understanding of mechanisms implicated in protection against canine infection. Nevertheless, most studies in dogs have not been carried out in organs that are targets of infection. This work is first to report the profile of cytokines and parasite burdens, as determined by real-time PCR, in the lymph nodes of dogs naturally infected with Leishmania chagasi. With this purpose, 18 mongrel dogs were divided in three groups: control non-infected dogs (n=6) and naturally infected animals with L. chagasi, asymptomatic (n=6) and symptomatic (n=6). Parasite burden in lymph nodes was 73-fold greater in symptomatic than asymptomatic animals. Prescapular lymph nodes of asymptomatic dogs had the highest expression of IFN gamma and TNF-alpha and low parasite burden, indicating that these cytokines play a role in protection against infection. Highest expression of IL-10 and TGF-beta and high parasite burden were observed in symptomatic dogs, suggesting a role for these cytokines in the progression of disease. Hence, the balance of expression of IFN-gamma and TNF-alpha (protective) and IL-10 and TGF-beta (disease progression) in lymph nodes determine parasite burden and clinical expression in naturally infected dogs. PMID- 19124161 TI - Juvenile nasopharyngeal angiofibroma: analysis of 42 cases and important aspects of endoscopic approach. AB - OBJECTIVE: To determine the general characteristics of 42 patients who were operated for juvenile nasopharyngeal angiofibroma (JNA); and to determine the important aspects and the advantages of endoscopic transnasal approach over other techniques. METHODS: Forty-two patients with JNA, 12 of whom were operated with endoscopic transnasal approach at a tertiary care center between March 1987 and February 2008 were evaluated retrospectively. The general characteristics of the patients, presenting signs and symptoms, the surgical approach performed, tumor stage, whether preoperative embolization was performed and rates of recurrence were studied. Patients who underwent surgery with endoscopic transnasal approach were compared to others who underwent surgery with different techniques. RESULTS: One patient was female and the remaining 41 patients were male. Twelve patients underwent endoscopic removal of the tumor. Lateral rhinotomy was used for 10 patients, degloving approach for 7 patients, transpalatal approach for 6 patients, combined approach for another 6 patients and finally midfacial splitting for 1 patient. Seventeen percent of the patients had referred with recurrent diseases. According to staging system of Radkowski, about half of the patients belonged to stage IIA or IIC. The mean age was 16 and the most common presenting symptom was nasal obstruction. Preoperative embolization rate was 59.5% while recurrence rate in the postoperative period was 17%. In non endoscopic approaches, the tumor contiguity and operative plan were determined via radiological data. In operations performed with endoscopic transnasal approach in patients with JNA, the radiological data as well as a detailed endoscopic tumor examination were utilized and it was observed that tumor invaded the middle turbinate in 67% and both the septum and middle turbinate in 42% of the patients. While preoperative embolization was performed only in larger tumors before 1998, it was recruited in all subjects after 1998. Preoperative and postoperative hemorrhage were less, and durations of packing and hospitalization were shorter in patients operated with endoscopic approach plus preoperative embolization. CONCLUSION: Endoscopic transnasal approach has advantages of no non cosmetic sequela, less hemorrhage and no disruption in facial skeleton. Besides, this method allows better visualization of tumor contiguity and enables dissection and ligature of vascular structures in JNA surgery. The rate of tumors associated with middle turbinate and septum is greater than that is assumed in patients with JNA. Starting the tumor excision with partial resection of the middle turbinate and subperiostal dissection of the septum and anterior sphenoidal wall, and avoiding direct contact with the tumor might decrease the amount of bleeding. PMID- 19124160 TI - Indices of inflammation in the lung and liver in the early stages of the black walnut extract model of equine laminitis. AB - The liver and lung are not only described as "target organs" in sepsis in most species, but are purported to be sources of circulating inflammatory mediators central to the systemic inflammatory response syndrome (SIRS). As we have recently reported an inflammatory response in the laminar tissue in laminitis similar to that described in "target organs" in human sepsis, we investigated the inflammatory response of the lung and liver in the black walnut extract (BWE) model of equine laminitis to determine (1) if a similar systemic inflammatory response occurs in this laminitis model as described for these organs in human sepsis, and (2) if these organs may be an important source of the inflammatory mediators leading to laminar inflammation. Real-time quantitative PCR (RT-qPCR) was used to measure hepatic and pulmonary mRNA concentrations of IL-1beta, IL-4, IL-6, IL-8, IL-10, TNF-alpha, COX-1 and COX-2. Hepatic samples were assessed from two time points in the developmental/prodromal period: (1) 1.5h post-BWE administration (BWE-1.5H, n = 5), and (2) the "developmental time point" (onset of leukopenia, approximately 3h post-BWE administration, BWE-DEV, n = 5). Pulmonary samples were only assessed for the BWE-DEV group. One control group (CON-3H, n = 5) was used for both the 1.5H and DEV groups. Finally, CD13 immunohistochemistry was performed to assess leukocyte emigration into hepatic and pulmonary parenchyma. Hepatic and pulmonary mRNA concentrations of the proinflammatory cytokines IL-6, IL-8 and TNF-alpha were significantly increased (P < 0.05) in BWE-1.5H and BWE-DEV groups compared to the control group; IL-1beta mRNA concentrations were only increased in the lung. The "anti-inflammatory" cytokines, IL-10 and IL-4, underwent transient decreases at different time points. Significant increases in parenchymal leukocyte numbers occurred in both the lung and liver at the BWE-DEV time point. Hepatic and pulmonary proinflammatory cytokine expression differ from that previously reported for the laminae in that TNF-alpha was increased in the hepatic and pulmonary tissues, the increases in expression of IL-6 and IL-8 are dramatically smaller for the liver and lung compared to those reported for the laminae, and the peak changes appear to occur later in the disease process in the liver than in the laminae (BWE-DEV in liver vs. 1.5H in the laminae). PMID- 19124162 TI - A new imaging modality in detection of caustic oesophageal injury: Technetium-99m pyrophosphate scintigraphy. AB - OBJECTIVES: Early oesophagoscopy is usually recommended in children after caustic ingestion to assess the severity of the initial digestive lesions. An oesophagoscopic procedure in children always demands to be performed under sedation and bears a certain risk of iatrogenic injury. The purpose of the study is to determine the value and sensitivity of Tc-99m pyrophosphate (99mTc-PYP) scintigraphy for the detection of caustic oesophageal injury. METHODS: The caustic oesophageal burns were created with 25% NaOH in an experimental rat model. Seventeen rats were divided into two groups. The BURN group (n=9) included caustic oesophageal burning rats. Non-BURN group (n=8) included sham-operated rats. All animals underwent 99mTc-PYP scintigraphy 72 h after the burning experiment. Anterior static and single-photon emission computed tomography images were obtained using a low-energy all-purpose collimator, 2.55 zoom factor, 3h after intravenous injection of 10 mCi 99mTc-PYP. After the imaging, all the oesophagi were explored for external determination of the injury sites and macroautoradiographic scintigraphy as well. Rectangular region of interest were placed in the visually determined lesion of oesophagus (O) as well as non-lesion area (N), and an ON ratio was created. The severity and extension of oesophageal burn were assessed 3 days after burning by histopathologic evaluation. The injuries were graded according to the ulcer depth, ulcer width, thrombus formation, and the development of perivascular fibrinoid necrosis. RESULTS: The oesophageal burning areas were demonstrated by visual evaluation of the Tc-99m PYP scintigraphy on all animals. The mean ON ratio of 99mTc-PYP was 21.61+/-7.01 in the BURN group and in the non-BURN group, was 2.25+/-0.24 (p<0.001). The best cut-off point of the ON ratio for caustic oesophageal injury was 5.45, with sensitivity and specificity of 100%. PYP scan findings were also confirmed by histological evidence of ulcer depth, ulcer width, thrombus formation, and perivascular fibrinoid necrosis in the same areas. CONCLUSIONS: The caustic oesophageal injuries have been demonstrated by 99mTc-PYP scintigraphy in burning experiment. 99mTc-PYP scintigraphy is sensitive for detection of caustic oesophageal injury. This new imaging tool is found to be sensitive and specific for the detection of caustic oesophageal injury in the rats. PMID- 19124163 TI - Benefits of the maximum tolerated dose (MTD) and maximum tolerated concentration (MTC) concept in aquatic toxicology. AB - There is increasing recognition of the need to identify specific sublethal effects of chemicals, such as reproductive toxicity, and specific modes of actions of the chemicals, such as interference with the endocrine system. To achieve these aims requires criteria which provide a basis to interpret study findings so as to separate these specific toxicities and modes of action from not only acute lethality per se but also from severe inanition and malaise that non specifically compromise reproductive capacity and the response of endocrine endpoints. Mammalian toxicologists have recognized that very high dose levels are sometimes required to elicit both specific adverse effects and present the potential of non-specific "systemic toxicity". Mammalian toxicologists have developed the concept of a maximum tolerated dose (MTD) beyond which a specific toxicity or action cannot be attributed to a test substance due to the compromised state of the organism. Ecotoxicologists are now confronted by a similar challenge and must develop an analogous concept of a MTD and the respective criteria. As examples of this conundrum, we note recent developments in efforts to validate protocols for fish reproductive toxicity and endocrine screens (e.g. some chemicals originally selected as 'negatives' elicited decreases in fecundity or changes in endpoints intended to be biomarkers for endocrine modes of action). Unless analogous criteria can be developed, the potentially confounding effects of systemic toxicity may then undermine the reliable assessment of specific reproductive effects or biomarkers such as vitellogenin or spiggin. The same issue confronts other areas of aquatic toxicology (e.g., genotoxicity) and the use of aquatic animals for preclinical assessments of drugs (e.g., use of zebrafish for drug safety assessment). We propose that there are benefits to adopting the concept of an MTD for toxicology and pharmacology studies using fish and other aquatic organisms and the development of sound criteria for data interpretation when the exposure of organisms has exceeded the MTD. While the MTD approach is well established for oral, topical, inhalational or injection exposure routes in mammalian toxicology, we propose that for exposure of aquatic organisms via immersion, the term Maximum Tolerated Concentration (MTC) is more appropriate. PMID- 19124164 TI - Effects of elevated temperature and cadmium exposure on stress protein response in eastern oysters Crassostrea virginica (Gmelin). AB - Stress proteins such as heat shock proteins (HSPs) and metallothioneins (MTs) play a key role in cellular protection against environmental stress. Marine ectotherms such as eastern oysters Crassostrea virginica are commonly exposed to multiple stressors including temperature and pollution by metals such as cadmium (Cd) in estuaries and coastal zones; however, the combined effects of these stressors on their cellular protection mechanisms are poorly understood. We acclimated C. virginica from populations adapted to different thermal regimes (Washington, North Carolina and Texas) at a common temperature of 12 degrees C, and analyzed their expression of MTs and HSPs (cytosolic HSP69, HSC72-77, HSP90 and mitochondrial HSP60) in response to the combined acute temperature stress and long-term Cd exposure. Overall, HSP and MT induction patterns were similar in oysters from the three studied geographically distant populations. HSP69 and MTs were significantly up-regulated by Cd and temperature stress implying their important role in cellular stress protection. In contrast, HSC72-77, HSP60 and HSP90 were not consistently induced by either acute heat or Cd exposure. The induction temperature for MTs was higher than for HSP69 (>28 degrees C vs. 20 degrees C, respectively), and MTs were more strongly induced by Cd than by temperature stress (to up to 38-94-fold compared by 3.5-7.5-fold, respectively) consistent with their predominant role in metal detoxification. Notably, heat stress did not result in an additional increase in metallothionein expression in Cd-exposed oysters suggesting a capacity limitation during the combined exposure to Cd and temperature stress. Levels of HSP69 and in some cases, HSC72-77 and HSP90 were lower in Cd-exposed oysters as compared to their control counterparts during heat stress indicating that simultaneous exposure to these two stressors may have partially suppressed the cytoprotective upregulation of molecular chaperones. These limitations of stress protein response may contribute to the reduced thermotolerance of oysters from metal-polluted environments. PMID- 19124165 TI - Effects of the azole fungicide Imazalil on the development of the ascidian Ciona intestinalis (Chordata, Tunicata): morphological and molecular characterization of the induced phenotype. AB - Imazalil (IMA) is a fungicide that is used extensively in fruit plantations and post-harvest treatments, but has teratogenic effects on vertebrate development, possibly due to the perturbation of retinoic acid (RA) levels in the embryo. Ascidians are sessile marine invertebrate chordates that develop through a tadpole larva, with a body plan that shares basic homologies with vertebrates. In this work, we tested the effects of IMA on the development of the solitary ascidian Ciona intestinalis by treating two-cell stage embryos with a range of concentrations (0.1, 0.5, 1, 2.5, 5, 10, 20 and 50microThe fungicide significantly altered ascidian development even at low concentrations and its effects were dose-dependent. Probit analysis revealed that the median lethal concentration, LC(50), was 4.87microM and the median teratogenic concentration, TC(50), was 0.73microM. Larvae developing from embryos exposed to IMA showed malformations of the anterior structures, which became more severe as IMA concentration increased. In particular, the anterior nervous system and the sensory vesicle were reduced, and the pigmented organs (the ocellus and the otolith) progressively lost their pigmentation. The larval phenotype induced by 5microM IMA exposure was further characterized by means of molecular analysis, through whole mount in situ hybridization with probes for genes related to the nervous system: Ci-Otp, Ci-GAD, Ci-POU IV, which are markers of the anterior neuro-ectoderm, the central nervous system and the peripheral nervous system respectively, and Ci-Hox-1, a gene specifically activated by RA, and Ci-Aldh2, a gene for aldehyde dehydrogenase, which is involved in RA synthesis. The altered expression of Ci-Otp, Ci-GAD, Ci-POU IV in 5microM IMA-exposed larvae compared to control larvae showed that this fungicide could affect the differentiation of the anterior nervous system, particularly of the sensory vesicle neurons. Recent studies suggest a similarity between IMA- and RA-induced phenotypes in tunicates, indicating that triazoles may also alter RA metabolism in ascidians. The observed Ci-Hox-1 and Ci-Aldh2 expression in control and treated larvae did not allow a direct link between IMA teratogenic potential and RA-dependent morphogenesis to be identified. It is likely that the fungicidal teratogenic mechanism involved RA signalling but that its effects on ascidian development depend on a more complex mechanism. PMID- 19124166 TI - PTH-analogs: comparable or different? AB - Because no comparative studies exist, no clear pronouncements can be made about the potential differences in effectiveness and safety between PTH 1-34 and PTH 1 84. As regards the efficacy, a convincing reduction of vertebral fractures was shown in both cases [Neer, R.M., Arnaud, C.D., Zanchetta, J.R., Prince, R., Gaich, G.A., Reginster, J.Y., Hodsman, A.B., Eriksen, E.F., Ish-Shalom, S., Genant, H.K., Wang, O., Mitlak, B.H., 2001. Effect of parathyroid hormone (1-34) on fractures and bone mineral density in postmenopausal women with osteoporosis. N. Engl. J. Med. 344, 1434-1441; Greenspan, S.L., Bone, H.G., Ettinger, M.P., Hanley, D.A., Lindsay, R., Zanchetta, J.R., Blosch, C.M., Mathisen, A.L., Morris, S.A., Marriott, T.B., Treatment of Osteoporosis with Parathyroid Hormone Study Group, 2007. Effect of recombinant human parathyroid hormone (1-84) on vertebral fracture and bone mineral density in postmenopausal women with osteoporosis: a randomized trial. Ann. Intern. Med. 146, 326-339]. A reduction of non-vertebral fractures was shown in the case of PTH 1-34 only. Another significant resemblance is that both medicines have a strong anabolic action; this mechanism of action is essentially different from the bisphosphonates and strontium ranelate. Both medicines constitute a welcome addition to the therapeutic arsenal for patients with severe osteoporosis. More data from literature (including information on follow-up data and use in men) are available for PTH 1-34 because it has been available for longer. As regards the side effect profile, PTH 1-84 appears to have a higher incidence of hypercalcemia, hypercalciuria and nausea than teriparatide. Here, too, no comparative study exists: the differences may therefore be based on an actual difference in side effects, or it may be ascribed to differences in definitions and/or patient populations. PMID- 19124167 TI - Home environmental problems and physical function in Taiwanese older adults. AB - Environmental hazards play an important role in the disablement process. The purpose of this study was to investigate the relationship between home environmental problems and personal physical function. Data were based on a two stage nationwide survey and evaluation on the needs of long-term care in Taiwan. A total of 10,596 individuals aged 65 and over were included in this study. These participants were identified with physical or cognitive problems at the screening interview and further evaluated at the second interview on health condition, functional status, needs of long-term care, and home environmental problems. Six items of environmental hazards were assessed at the participants' homes with direct observation. The prevalence rates of home environmental problems were similar among older adults with different levels of physical function. No grab bars (79.6-85.1%) and no protections against slip (81.9-92.8%) in the bathroom were two commonly present hazards in older adults' homes. Older adults with a higher income (Odds ratio=OR=0.75), without income information (OR=0.78) or living with other persons (OR=0.74) were less likely to experience environmental problems at home. Results from this study revealed that home environment condition was associated with factors other than personal disabling conditions for the elderly. Modifying home environment, especially the bathroom, should be attached with great importance for physically disabled older adults. PMID- 19124168 TI - Investigation on status and influential factors of cognitive function of the community-dwelling elderly in Changsha City. AB - The present study describes the results of investigations on the cognitive status and the influential factors of cognitive function in the community-dwelling elderly of Changsha City, China. Analyzing data of 1000 subjects over 60 years of age was collected by the self-designed demographic information sheet, the Mini Mental State Examination (MMSE) and Activity of Daily Living (ADL) scales. The result showed that MMSE mean score was 24.35+/-4.91, ranging from 4 to 30; there were 275 subjects with cognitive impairments of various extents. These results reveal that the elderly in this study differed quite strongly in their cognitive function; over one-fourth of them showed slight to severe cognitive impairment. Moreover, the influential factors such as gender, age, marriage status, education level, occupation, income, hobby, caregivers, medical history, self-appraised health, and ADL were detected. Of these parameters, age, education level, gender, self-appraised health, ADL, medical history, and caregivers were the striking influential factors. PMID- 19124169 TI - Serum and intracellular magnesium deficiency in patients with metabolic syndrome- evidences for its relation to insulin resistance. AB - This cross sectional study evaluated serum (SMg) and intramononuclear (MMg) magnesium in patients with metabolic syndrome without diabetes and correlated them with cardiovascular risk factors. 72 patients and 57 controls (blood donors) were studied. Hypomagnesemia (SMg<1.7 mg/dL) was seen in 23.2% and intracellular depletion in 36.1% of the patients. SMg and MMg means were significantly lower in patients than in controls: 1.80+/-0.18 mg/dL vs. 2.43+/-0.43 mg/dL and 0.98+/ 0.55 microg/mg vs. 1.67+/-0.64 microg/mg of protein (P<0.001). Inverse correlation was observed between, SMg and MMg with BMI; SMg with systolic blood pressure and waist circumference in women. Patients with acanthosis nigricans had lower SMg (1.75+/-0.18 mg/dL vs. 1.85+/- 0.18 mg/dL, P<0.05). Non-white people had lower SMg (1.78+/-0.16 mg/dL vs. 1.92+/-0.24 mg/dL, P=0.007) and MMg (0.95+/ 0.59 microg/mg vs. 1.13+/-0.42 microg/mg, P=0.03). Patients with IR showed lower MgM means (0.84+/-0.33 microg/mg vs. 1.14+/-0.69 microg/mg, P<0.05). The same occurred in patients with low HDL-c levels (0.92+/-0.46 microg/mg vs. 1.20+/-0.70 microg/mg, P=0.03), and those with moderate and severe hepatic steatosis (0.77+/ 0.29 microg/mg vs. 1.21+/-0.80 microg/mg, P<0.05). In conclusion, magnesium depletion in serum and mononuclear cells is common in obese people with metabolic syndrome, and it is more evident in non-white people with insulin resistance. This depletion may contribute to a post-receptor insulin resistance. PMID- 19124170 TI - The status of glycemic control by general practitioners and specialists for diabetes in Japan: a cross-sectional survey of 15,652 patients with diabetes mellitus. AB - To determine the status of diabetes care by general practitioners and diabetes specialists in Japan, we conducted a nation-wide cross-sectional survey. We asked 8112 clinics and hospitals randomly, from throughout Japan, to participate in this study and 721 facilities agreed. A total of 15,652 patients aged from 15 to 97 with type 1 and type 2 diabetes were enrolled. Of these, 14,560 (93.0%) and 1092 (7.0%) patients were cared for by general practitioners and diabetes specialists, respectively. HbA1c levels were measured by a latex agglutination method, and age, height, body weight, type of diabetes and treatment modality were obtained from each patient. Mean HbA1c level for all patients treated by general practitioners was significantly lower than for those treated by the diabetes specialists (6.8+/-1.2% vs. 7.0+/-1.2%, p=0.0002). Mean HbA1c level for patients without insulin therapy was lower than for those treated with insulin, irrespective of caring physician. The proportion of patients treated with insulin therapy by diabetes specialists was higher (17.7%) than that by general practitioners (6.5%). This study showed that average HbA1c levels in Japanese patients treated by either general practitioners or specialists was acceptable, regardless of study limitations or bias. PMID- 19124172 TI - Hyperglycaemia following glucose challenge test during pregnancy: when can a screening test become diagnostic? AB - OBJECTIVE: The 50g-glucose challenge test (GCT) is commonly used for screening of gestational diabetes (GDM) in low risk pregnant women. If elevated, glucose tolerance test is performed to confirm the diagnosis. In this study, we evaluated whether GCT alone is sufficient to diagnose GDM when the GCT result is very elevated. RESEARCH DESIGN AND METHODS: Using a database of 62877 pregnancies over 10 years, the positive predictive value (PPV) of GCT for GDM was assessed using different GCT cut-off values. RESULTS: At a glucose cut-off value of 11 mmol/l, the PPV for GDM was 85.3%, based on the subsequent GTT. This increased to 95.3% when the cut-off was 12 mmol/l. Furthermore, the PPV was consistently higher when GCT was performed in the morning. CONCLUSION: We concluded that the diagnosis of GDM can be made when the glucose level following GCT is very elevated, and GTT need not to be performed for confirmation of GDM. The timing of GCT also affected PPV for GDM, and has implications on the diagnostic value of the test. PMID- 19124171 TI - Classical PKC is not associated with defective insulin signaling in patients with impaired glucose tolerance. AB - BACKGROUND AND AIM: To investigate the role of insulin signaling defects in impaired glucose tolerance (IGT), we assessed the functionality of the insulin signaling cascade before and after insulin stimulation in both IGT group and control group. METHODS: Ten IGT subjects and 15 control subjects were recruited for this study. Whole-body insulin-mediated glucose uptake was determined using a euglycemic hyperinsulinemic clamp test. Muscle biopsies were obtained from the vastus lateralis muscle before and after insulin stimulation, to assess the insulin signaling cascade. RESULTS: The insulin-stimulated incremental changes in phosphorylated IR-beta, IRS, Akt, and GSK-3 beta and in the membrane-associated PKC-zeta protein level were reduced in the IGT group compared with those in the control group (p<0.05). The membrane-associated PKC-lambda protein level was also reduced in the IGT group, but not significantly so (p=0.08). The incremental changes in the protein levels of PKC-alpha, -beta, and -theta were not significantly different between the two groups. CONCLUSION: The subjects with IGT showed decreased membrane-associated PKC-zeta/lambda activity in response to insulin stimulation, as well as defects in early insulin signaling. Our results suggest that membrane-associated PKC-alpha and -beta may not be associated with insulin resistance in IGT. PMID- 19124173 TI - Clustering dynamic PET images on the Gaussian distributed sinogram domain. AB - Segmentation of dynamic PET images is an important preprocessing step for kinetic parameter estimation. The time activity curve (TAC) of individual pixels has very low signal-to-noise ratio (SNR). Therefore, the kinetic parameters estimated from the TAC of an individual pixel may not be accurate, and these estimations may have very high spatial variance. To alleviate this problem, pixels with similar kinetic characteristics are clustered into regions, and TACs of pixels within each region are averaged to increase SNR. It has recently been shown that clustering dynamic PET images in the sinogram domain is better than clustering them in the reconstructed image domain [M.E. Kamasak, B. Bayraktar, Clustering dynamic PET images on the projection domain, IEEE Trans. Nucl. Sci. 54 (3) (June 2007) 496-503.]. In that study, the sinograms are assumed to have Poisson distribution. The clusters and TACs of the clusters are then chosen to maximize the posterior probability of the measured sinograms. Although the raw sinogram data are Poisson distributed, the sinogram data corrected for scatter, randoms, attenuation etc. are not Poisson distributed anymore. In this paper, we describe how to cluster dynamic PET images on the sinogram domain when the sinograms are Gaussian distributed. PMID- 19124174 TI - Culture and functional studies of mouse macrophages on native-like fibrillar type I collagen. AB - Freshly isolated, starch-elicited mouse peritoneal macrophages (Mo) attached very efficiently to type I collagen in vitro, if collagen molecules were arranged in ordered supra-molecular assemblies corresponding to the precursor native fibrils. After 6-20h of incubation, the collagen-bound cells were observed to secrete fibronectin, which presumably enhanced cell-collagen interaction associated with cellular differentiation. Mo attachment to collagen could be temporarily inhibited by addition of the linear tri-peptide Arg-Gly-Asp (RGD) to the culture media. This inhibition was much more pronounced when using the cyclic RGD containing peptide cGRGDSPA. Similarly, cells could be easily detached from the fibrillar collagen layers within 20 min at 37 degrees C by RGDS, GRGDS or cGRGDSPA but not by the glutamate-containing RGES peptide. Using antibodies to known collagen receptors, attachment of Mo to type I collagen fibers was best inhibited by antibodies directed against the alpha2 and beta1 integrin subunits. The presence of these integrins on Mo was confirmed by immunofluorescence. Binding of the alpha2beta1 integrin on collagen was divalent cation-dependent and was supported by magnesium but not by calcium. Cells recovered by RGD-mediated detachment from collagen were highly phagocytic and synthesized DNA when exposed to growth factors. These cells could be activated for cytotoxicity by treatment with interferon-gamma and lipopolysaccharide. Comparative in vitro assays performed on macrophages cultured on plastic and on collagen allowed the detection of NO production by activated macrophages followed by spontaneous deactivation for cells cultivated on collagen. These findings suggest that Mo can recognize native collagen of type I through functional interactions with their specific triple helix-binding integrin receptors indicating that integrins other than those directed to fibronectin may also occupy active focal points on the cell at the initial phase of attachment. PMID- 19124176 TI - Developing inter-professional learning: tactics, teamwork and talk. AB - SUMMARY: Teamwork and collaboration between all health professionals results in high quality clinical care, and increased job satisfaction for staff. Encouraging inter-professional learning (IPL) may be advantageous in developing more effective teams. There is little rigorous research in this area, but many small uncontrolled studies do demonstrate positive results. IPL involves structured learning opportunities that enhance problem-solving abilities and conflict resolution. It should be clearly differentiated from shared teaching (or multidisciplinary/multiprofessional learning), where common content is taught to many professions without any intention to develop interaction. To counteract the sometimes negative attitudes in both students and staff, educators need to commence IPL early in the programme, base it in both theoretical and clinical placements and ensure that it is valued and assessed. Difficulties with timetabling and accommodation need to be solved prior to commencement. A facilitator should be employed, and a team of committed lecturers developed, with an emphasis on teamwork and the discouragement of individualism. Opportunities for student interaction and ways of improving group dynamics within non threatening learning environments should to be sought, and instances of conflict embraced and resolved. Future IPL programmes should be rigorously evaluated and may demonstrate enhanced inter-professional relationships and improved quality of patient/client care. PMID- 19124177 TI - A national review of mandibular orthognathic surgery activity in the National Health Service in England over a nine year period: part 2--patient factors. AB - We aimed to investigate patients who had mandibular orthognathic surgery in England for the period 1 April 1997-30 March 2006. Data on hospital inpatient activity were extracted from the NHS Hospital Episode Statistics (HES) database for all mandibular orthognathic surgical procedures between these dates. Details of patients were investigated in relation to clinical details, socioeconomic status, other diagnoses, and clinical outcomes. Mandibular orthognathic operations were done for 8941 patients in England over the 9-year period, and the ratio of women: men was 1.7:1. The mean (SD) age at operation was 24 (9) years with no difference between sexes. There was a socioeconomic gradient in patients requesting treatment, most of whom lived in relatively affluent areas. The most common other diagnosis was "additional dentofacial anomalies" (8% of episodes). Mental or behavioural diagnoses were recorded in 41 patients (<1%). One patient died in hospital. Our results confirmed the differences in sex in patients receiving orthognathic care and highlighted socioeconomic inequalities in the treated patients. A number of coexisting diagnoses were also identified. PMID- 19124175 TI - Antioxidants have a rapid and long-lasting effect on neuritic abnormalities in APP:PS1 mice. AB - Senile plaques are a major pathological hallmark of Alzheimer's disease (AD). Compelling evidence suggests that senile plaques lead to structural alterations of neuronal processes and that local toxicity may be mediated by increased oxidative stress. Anti-oxidant therapy can alleviate the neuronal abnormalities in APP mice, but the time-course of this beneficial effect is unknown. We used multiphoton microscopy to assess in vivo the characteristics of antioxidant treatment on senile plaques and neurites in AD model mice (APPswe/PS1dE9). We observed that alpha-phenyl-N-tert-butyl nitrone (PBN), Ginkgo biloba extract (EGb 761) and Trolox had no effect on the size of existing senile plaques. However, all anti-oxidants had a straightening effect on curved neurites. This effect was detected as soon as 4 days after commencing the treatment, and was maintained after 1 month of daily treatment, with no further increase in the effect. The straightening of neurites persisted 15 days after stopping the treatment. These data indicate that neuronal plasticity is fast and still active in adult animals, and suggest that amelioration of the neuritic distortions associated with senile plaques with antioxidants is both rapid and long lasting. PMID- 19124178 TI - A national review of mandibular orthognathic surgery activity in the National Health Service in England over a nine year period: part 1--service factors. AB - We aimed to investigate the changing provision of mandibular orthognathic surgery in England for the period 1 April 1997-30 March 2006. Data on hospital inpatient activity were extracted from the NHS Hospital Episode Statistics (HES) database for all mandibular orthognathic surgical procedures between these dates. We investigated provider factors (in which units providing services were classified as undertaking either high or low volumes of activity) and temporal changes. There was a steady increase in the number of procedures over time, and a decrease in the mean time spent in hospital (p<0.001). Provider units with high volumes of activity had shorter inpatient stays than those that provided low volume (p<0.001) and exhibited a greater reduction in the duration of inpatient stays (p=0.02). Patterns of care are changing, and increasing numbers of treatments are being done as day-cases (p<0.001). Units with high volumes seem to be more efficient in terms of duration of hospital stay than low volume units. However, our data do not allow an insight into the quality of care provided, and further research is needed to address this issue. PMID- 19124179 TI - Does invasion of oral squamous cell carcinoma occur into a non-pathological fracture of the mandible? AB - Although the route of invasion of oral squamous cell carcinoma into the intact mandible is well-documented we know of no reports about whether malignant cells invade through non-pathological fractures. A 68-year-old patient presented with a traumatic fracture of the mandible at the site of an existing carcinoma. He had asegmental resection but histological examination showed no evidence of invasion into the bone. PMID- 19124180 TI - Re: Reconstruction of curved mandibular angle defects using a new internal transport distraction device: an experiment in goats. PMID- 19124181 TI - Particle-size distribution and gas/particle partitioning of atmospheric polybrominated diphenyl ethers in urban areas of Greece. AB - Ambient concentrations, gas/particle partitioning and particle-size distribution of polybrominated diphenyl ethers (PBDEs) were investigated in two urban areas (Athens and Heraklion) of Greece. Atmospheric (gas+particle) concentrations of summation operatorPBDE varied from 21 to 30 pg m(-3) in the center of Athens and from 4 to 44 pg m(-3) in the suburbs of Heraklion. A predominance of particulate PBDEs was observed in Athens (71-76% in particles), whereas the opposite was evident in Heraklion (69-92% in gas phase). In both urban areas, PBDE particle size distribution featured a distinct enrichment in smaller particles. A similar trend was also observed in aerosols of a background marine site. For all sampling sites, more than 46% of summation operatorPBDE was associated with particles of <0.57 microm in diameter. Our results imply that particulate PBDEs may have long atmospheric residence time and they may be capable of reaching the deeper parts of the human respiratory system. PMID- 19124182 TI - Removal of trace elements in three horizontal sub-surface flow constructed wetlands in the Czech Republic. AB - Between March 2006 and June 2008 removal of 34 trace elements was measured on a monthly basis at three horizontal-flow constructed wetlands in the Czech Republic designed to treat municipal wastewater. In general, the results indicated a very wide range of removal efficiencies among studied elements. The highest degree of removal (average of 90%) was found for aluminum. High average removal was also recorded for zinc (78%). Elements removed in the range of 50-75% were uranium, antimony, copper, lead, molybdenum, chromium, barium, iron and gallium. Removal of cadmium, tin, mercury, silver, selenium and nickel varied between 25 and 50%. Low retention (0-25%) was observed for vanadium, lithium, boron, cobalt and strontium. There were two elements (manganese and arsenic) for which average outflow concentrations were higher compared to inflow concentrations. Reduced manganese compounds are very soluble and therefore they are washed out under anaerobic conditions. PMID- 19124183 TI - Mapping a non-carious cervical lesion using stereoimagery and dental casts incorporating optical texture. AB - OBJECTIVES: Recent advances in digital photogrammetry have enabled a new approach to high resolution mapping of tooth surfaces. The aim of this study was to assess the accuracy and the precision of a new system using automatic digital stereo photogrammetry combined with an experimental casting material and to measure tooth surface loss in a non-carious cervical lesion. METHODS: A test object and tooth replicas incorporating optical texture obtained at baseline, one and 2 years were imaged with a pre-calibrated stereocamera. The stereoimagery was then processed with digital photogrammetric software to automatically generate digital surface models. Test object models were analysed for accuracy and precision and tooth surface models were aligned with point cloud analysis software and tooth surface loss determined. RESULTS: Replicas were mapped to an accuracy of (6+/ 13)microm. The sensitivity of change detection on tooth replicas was 40 microm with change ranging from 40 to 70 microm per annum in a discrete part of the lesion. CONCLUSIONS: In the case examined, the change detection data were of sufficient accuracy and resolution to draw meaningful conclusions about the spatial distribution and quantum of tooth hard tissue loss. The results of this investigation suggest that annual change detection studies will provide a clearer picture of the pattern of tooth surface loss and, in combination with other analytical techniques, a more detailed explanation of the natural history of these lesions. PMID- 19124184 TI - In vitro evaluation of casein phosphopeptide-amorphous calcium phosphate (CPP ACP) effect on stripped enamel surfaces. A SEM investigation. AB - OBJECTIVES: Interproximal enamel reduction (orthodontic stripping) leaves surface roughness that encourages plaque adherence. Sealants and fluoride products have been recommended to avoid possible detrimental effects. The objective of this study was to qualitatively evaluate, by means of scanning electron microscopy (SEM), the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on stripped enamel morphology after exposure to an acid solution. METHODS: 15 extracted lower incisors underwent stripping with metal strips (Horico, 80 microm) by 1 operator. Mesial and distal surfaces were sectioned into 30 samples (22 stripped; 8 unstripped) and assigned to 2 groups: Group A (2 stripped; 2 unstripped) served as control; in Group B (20 stripped; 6 unstripped) tooth mousse containing CPP-ACP was tested. For 8 days, teeth were placed in acid lactic solution for 2h three times a day (from 9a.m. to 9p.m.) with 2h water preservation in between. After each demineralizing bath, samples of group B were coated with CPP-ACP for 5 min before immersion into water. RESULTS: In Group A, stripped samples exhibited greater demineralization compared to unstripped. In Group B, CPP-ACP reduced enamel dissolution on both intact and abraded samples compared to Group A. CONCLUSIONS: Injuries to enamel morphology caused by stripping might predispose to greater demineralization in the event of exposure to acid environment. Topical applications of CPP-ACP could be effective in promoting enamel remineralization after interdental stripping. PMID- 19124185 TI - Influence of glutaraldehyde priming on bond strength of an experimental adhesive system applied to wet and dry dentine. AB - OBJECTIVES: This study tested the following null hypotheses: (1) there is no difference in resin-dentine bond strength when an experimental glutaraldehyde primer solution is added prior to bonding procedures and (2) there is no difference in resin-dentine bond strength when experimental glutaraldehyde/adhesive system is applied under dry or wet demineralized dentine conditions. METHODS: Extracted human maxillary third molars were selected. Flat, mid-coronal dentine was exposed for bonding and four groups were formed. Two groups were designated for the dry and two for the wet dentine technique: DRY: (1) Group GD: acid etching+glutaraldehyde primer (primer A)+HEMA/ethanol primer (primer B)-under dried dentine+unfilled resin; (2) Group D: the same as GD, except for primer A application; WET: (3) Group GW: the same as GD, but primer B was applied under wet dentine condition; (4) Group W: the same as GW, except for primer A application. The bonding resin was light-cured and a resin core was built up on the adhesive layer. Teeth were then prepared for microtensile bond testing to evaluate bond strength. The data obtained were submitted to ANOVA and Tukey's test (alpha=0.05). RESULTS: Glutaraldehyde primer application significantly improved resin-dentine bond strength. No significant difference was observed when the same experimental adhesive system was applied under either dry or wet dentine conditions. These results allow the first null hypothesis to be rejected and the second to be accepted. CONCLUSION: Glutaraldehyde may affect demineralized dentine properties leading to improved resin bonding to wet and dry substrates. PMID- 19124186 TI - Occlusal caries: Evaluation of direct microscopy versus digital imaging used for two histological classification systems. AB - OBJECTIVE: Histology is frequently used as a gold standard to validate caries detection devices. Poor assessment consistency could lead to apparent changes in diagnostic accuracy. In multi-center, multi-examiner studies electronic transfer of information would be convenient, provided there is no deteriation in quality. This study tested the hypothesis that examiner reproducibility in the assessment of caries lesion depth when viewing photographic images of histological sections on a computer monitor, is comparable with viewing the same sections under a microscope using two histological classification systems. METHODS: 166 investigation sites (96 teeth) were selected for visual examination (ICDAS-II) and sections made using a novel technique which reduced risk of section damage and allowed accurate allocation of section to each investigation site. Digital images of the sections were produced and four examiners viewed the sections under a microscope and on a separate occasion corresponding digital images on a computer monitor. Presence and extent of caries was scored according to two histological classification systems (Downer, ERK). RESULTS: The inter- and intra examiner reproducibility for both histological classification systems and both examination techniques was substantial to almost perfect (weighted kappa=0.63 0.90). Comparing the kappa values between microscopy and viewing digital images, there was no effect or only a small effect between both examination techniques (effect size 0.00-0.28). There was also a strong relationship between the two viewing techniques (r(s)=0.748-0.844). CONCLUSIONS: Viewing digital images of tooth sections produces results comparable to viewing images directly under a microscope and therefore has potential benefits for multi-centre studies. PMID- 19124187 TI - Assessing fluoride treatment and resistance of dental enamel to soft drink erosion in vitro: applications of focus variation 3D scanning microscopy and stylus profilometry. AB - OBJECTIVES: This study assesses the application of the focus variation 3D microscopy for the evaluation of dental erosion and fluoride treatment for prevention of enamel erosion in vitro. METHODS: Human dental enamel disks were treated with Prevident 5000 (PV, n=15) for 1 week and compared with a reference group (PN, n=15) after orange juice erosion in vitro. A focus variation 3D scanning microscope (IFM) and a stylus type profilometer (SSP) were used to evaluate the erosion depths on enamel. 3D topographic images were taken with vertical resolutions of 0.1 and 0.02 microm. Scratch marks depths from SSP were measured on IFM images. Measurements were compared between the SSP and IFM and between the two study groups. RESULTS: The SSP and IFM measurements of eroded enamel surfaces showed similar trends between the two methods and between the two study groups. The SSP and the IFM measurements were statistically significantly different but correlated with each other. PV group showed consistently lower erosion depth than PN in all profile measures using both SSP and IFM. The stylus tip created scratch marks that were significantly different in depths between the eroded and the reference surfaces in both groups. CONCLUSIONS: The focus variation 3D microscopy is a powerful tool in evaluating surface topography associated with enamel erosion and in assessing the treatment effects of anti erosive therapies. Topical treatment with Prevident 5000 significantly increased enamel resistance to erosion by orange juice and should be considered as a treatment choice in patients susceptible to acidic dental erosion. PMID- 19124188 TI - Comment on Spallicci et al. "Use of hyaluronidase for cervical ripening: a randomized trial" [Eur. J. Obstet. Gynecol. Reprod. Biol. 130 (1) (2007) 46-50]. PMID- 19124189 TI - Is metastasectomy for urothelial carcinoma worthwhile? PMID- 19124190 TI - New onset of idiopathic bilateral ear tics in an adult. AB - Tic disorders are commonly considered to be childhood syndromes. Newly presenting tic disorders during adulthood are uncommon and mostly described in relation to an acquired brain lesion or as incidental tics, particularly in context with other neurological or psychiatric diseases. Tic disorder involving the ears is extremely uncommon with only few studies in English literature. In the present case, we describe an adult patient with new-onset idiopathic tics disorder involving both ears, causing social embarrassment. In addition, our patient had recent onset of the tics without any childhood or family history of tic disorders. The single most important component of management is an accurate diagnosis. At the same time, tics should be differentiated from other movement disorders such as chorea, stereotypy, and dystonias. PMID- 19124191 TI - Adsorption of Cr(VI) onto cationic surfactant-modified activated carbon. AB - Highly toxic oxyanions, such as hexavalent chromium (Cr(VI)), have caused adverse effects on human health. This study evaluated the feasibility of using cationic surfactant-modified activated carbon (AC) to remove Cr(VI). To modify activated carbon using a cationic surfactant, AC was mixed with a surfactant solution of 0.5 critical micelle concentration (CMC), at which concentration the surfactant molecules exist as mono-molecules. Adsorption kinetics and an isotherm model were used to study the adsorption characteristics of Cr(VI) onto MAC. The adsorption capacity of MAC for Cr(V) was enhanced compared to that of AC. MAC modified by hexadecyltrimethylammonium had a higher adsorption capacity for the removal of Cr(VI) than that modified by cetylpyridinium. The modification of AC by a cationic surfactant enhanced both its Cr(VI) adsorption rate and its Cr(VI) adsorption capacity. The breakthrough point of MAC for Cr(VI) was 100 times greater than for the raw AC. As a result, MAC is a promising adsorbent to treat Cr(VI) in an aqueous stream. PMID- 19124192 TI - Effect of aggregate structure on VOC gas adsorption onto volcanic ash soil. AB - The understanding of the gaseous adsorption process and the parameters of volatile organic compounds such as organic solvents or fuels onto soils is very important in the analysis of the transport or fate of these chemicals in soils. Batch adsorption experiments with six different treatments were conducted to determine the adsorption of isohexane, a gaseous aliphatic, onto volcanic ash soil (Tachikawa loam). The measured gas adsorption coefficient for samples of Tachikawa loam used in the first three treatments, Control, AD (aggregate destroyed), and AD-OMR (aggregate destroyed and organic matter removed), implied that the aggregate structure of volcanic ash soil as well as organic matter strongly enhanced gas adsorption under the dry condition, whereas under the wet condition, the aggregate structure played an important role in gas adsorption regardless of the insolubility of isohexane. In the gas adsorption experiments for the last three treatments, soils were sieved in different sizes of mesh and were separated into three different aggregate or particle size fractions (2.0 1.0mm, 1.0-0.5mm, and less than 0.5mm). Tachikawa loam with a larger size fraction showed higher gas adsorption coefficient, suggesting the higher contributions of macroaggregates to isohexane gas adsorption under dry and wet conditions. PMID- 19124193 TI - Synergistic effect of catalyst for oxidation removal of toluene. AB - A series of experiments was performed for toluene removal from a gaseous influent at the normal temperature and atmospheric pressure by decomposition due to dielectric barrier discharge generated non-thermal plasma, by using MnO(2)/gamma Al(2)O(3) as catalyst. The removal efficiency of toluene was significantly increased by combining MnO(2)/gamma-Al(2)O(3) with NTP. At the same time, the goal of improving energy efficiency and decreasing O(3) from exhaust gas treatment was accomplished. PMID- 19124194 TI - Preparation of a new sorbent with hydrated lime and blast furnace slag for phosphorus removal from aqueous solution. AB - The removal of dissolvable inorganic phosphate (H(2)PO(4)(-)) by sorbents prepared from hydrated lime (HL) and blast furnace slag (BFS) was fundamentally studied by an orthogonal experiment design. Based on statistic analysis, it is revealed that the weight ratio of BFS/HL is the most significant variable, and an optimized preparation condition is figured out. With the increase of HL content, the adsorption capacity increases, suggesting that the HL plays the important role in the removal process in the gross. However, in the lower HL content, it is interesting that the adsorption capacity of as-prepared sorbents exceed the sum of the capacities of the same ratio of BFS and HL. The further analysis indicate the excess capacities linearly depend on the specific surface area of sorbents, suggesting that the removal of H(2)PO(4)(-) is closely related with the microstructure of sorbents in the lower HL content, according to the characterization with SEM, XRD and pore analysis. Additionally, an adsorption model and kinetic are discussed in this paper. PMID- 19124195 TI - Catalytic ozonation of dimethyl phthalate and chlorination disinfection by product precursors over Ru/AC. AB - Catalytic ozonation of dimethyl phthalate (DMP) in aqueous solution (5mg/L) under various reactions was performed to examine the effect of catalyst dosage, catalyst particle size, ozone dosage, and gas flow rate on the mineralization of DMP. The mineralization of DMP can be achieved via ozonation and the presence of Ru/AC could greatly accelerate the mineralization rate of DMP in ozonation process. In the continuous experiment of the Ru/AC catalyzed ozonation of DMP, total organic carbon (TOC) removals were kept stable around 75% during 42 h reaction. No leaching of ruthenium was observed in the treated water samples. The treatment of natural water using Ru/AC+O(3), Ru/AC+O(2) and ozonation alone was studied. In the Ru/AC+O(3) process, TOC removals, the reductions of the haloacetic acid formation potentials (HAAFPs), and the reductions of the trihalomethane formation potentials (THMFPs) of 11 water samples were 22-44%, 39 61% and 50-65%, respectively. Ru/AC+O(3) process was much more effective than ozonation alone for TOC removal and the reduction of disinfection by-product formation potential (DBPFP) in the treatment of natural water. It is a promising water treatment technology. PMID- 19124196 TI - Environmentally friendly efficient coupling of n-heptane by sulfated tri component metal oxides in slurry bubble column reactor. AB - SO(4)(2-)/M(x)O(y) is of the greatest interest in solid catalysts and green catalysts. Slurry bubble column reactors are of considerable interest in industrial processes and various biochemical processes. The cetane number (CN) has widely used diesel fuel quality parameter related to the ignition delay time (and combustion quality) of a fuel. CN improvement of diesel fuels is a difficult task that refiners will face in the near future. For that purpose, the tests were designed in which n-heptane is used as the reactant in the air or ozone atmosphere at room temperature (RT) and local atmospheric pressure (LAP) using different catalysts of sulfated tri-component metal oxides SO(4)(2-)/Fe(2)O(3) TiO2-SnO(2) (SFTSn) and SO(4)(2-)/MnO(2)-TiO2-SnO(2) (SMTSn) in slurry bubble column reactor. The products distribution was analyzed by gas chromatography-mass spectrometry (GC-MS) method and the results show that the relative selectivity of long linear alkane (C(12)-C(28)) reaches the maximum (87.330%) when SMTSn is used as catalyst in flow air at 60 min. Diesel fuel components with higher cetane numbers can be easily obtained from this study. PMID- 19124198 TI - An experimental study on the hazard assessment and mechanical properties of porous concrete utilizing coal bottom ash coarse aggregate in Korea. AB - This study evaluates quality properties and toxicity of coal bottom ash coarse aggregate and analyzes mechanical properties of porous concrete depending on mixing rates of coal bottom ash. As a result, soundness and resistance to abrasion of coal bottom ash coarse aggregate were satisfied according to the standard of coarse aggregate for concrete. To satisfy the standard pertaining to chloride content, the coarse aggregates have to be washed more than twice. In regards to the result of leaching test for coal bottom ash coarse aggregate and porous concrete produced with these coarse aggregates, it was satisfied with the environment criteria. As the mixing rate of coal bottom ash increased, influence of void ratio and permeability coefficient was very little, but compressive and flexural strength decreased. When coal bottom ash was mixed over 40%, strength decreased sharply (compressive strength: by 11.7-27.1%, flexural strength: by maximum 26.4%). Also, as the mixing rate of coal bottom ash increased, it was confirmed that test specimens were destroyed by aggregate fracture more than binder fracture and interface fracture. To utilize coal bottom ash in large quantities, it is thought that an improvement method in regards to strength has to be discussed such as incorporation of reinforcing materials and improvement of aggregate hardness. PMID- 19124197 TI - Evaluation of arsenic and other physico-chemical parameters of surface and ground water of Jamshoro, Pakistan. AB - Arsenic contamination in water has caused severe health problems around the world. The purpose of this study was to evaluate the geological and anthropogenic aspects of As pollution in surface and groundwater resources of Jamshoro Sindh, Pakistan. Hydride generator atomic absorption spectrophotometry (HG-AAS) is employed for the determination of arsenic in water samples, with detection limit of 0.02 microg l(-1). Arsenic concentrations in surface and underground water range from 3.0 to 50.0, and 13 to 106 microg l(-1), respectively. In most of the water samples As levels exceeded the WHO provisional guideline values 10 microg l(-1). The high level of As in under study area may be due to widespread water logging from Indus river irrigation system which causes high saturation of salts in this semi-arid region and lead to enrichment of As in shallow groundwater. Among the physico-chemical parameters, electrical conductivity, Na(+), K(+), and SO(4)(2-) were found to be higher in surface and ground water, while elevated levels of Ca(2+) and Cl(-) were detected only in ground water than WHO permissible limit. The high level of iron was observed in ground water, which is a possible source of As enrichment in the study area. The multivariate technique (cluster analysis) was used for the elucidation of high, medium and low As contaminated areas. It may be concluded that As originate from coal combustion at brick factories and power generation plants, and it was mobilized promotionally by the alkaline nature of the understudy groundwater samples. PMID- 19124199 TI - Gold(III) biosorption and bioreduction with the brown alga Fucus vesiculosus. AB - In this paper, the bioreduction of Au(III) to Au(0) using biomass of the brown alga Fucus vesiculosus was investigated. The recovery and reduction process took place in two stages with an optimum pH range of 4-9 with a maximum uptake obtained at pH 7. In the first stage, an induction period previous to gold reduction, the variation of pH, redox potential and gold concentration in solution was practically negligible and no color change was observed. In the second stage, the gold reduction was followed by a sharp decrease of gold concentration, pH and redox potential of solution and a color change from yellow to reddish purple. Hydroxyl groups present in the algal polysaccharides were involved in the gold bioreduction. Metallic gold was detected as microprecipitates on the biomass surface and in colloidal form as nanoparticles in the solution. Bioreduction with F. vesiculosus could be an alternative and environmentally friendly process that can be used for recovering gold from dilute hydrometallurgical solutions and leachates of electronic scraps, and for the synthesis of gold nanoparticles of different size and shape. PMID- 19124200 TI - Octadecyl bonded silica membrane disk modified with Cyanex302 for separation and flame atomic absorption spectrometric determination of nickel from tap water and industrial effluent. AB - A simple and reliable method based upon impregnation of Cyanex302 on octadecyl bonded silica membrane disk has been developed for separation and atomic absorption spectrometric determination of nickel. The influence of various parameters like aqueous phase pH, flow rate and volume of eluent were investigated systematically to optimize the conditions for quantitative sorption and desorption of nickel. The break through volume for nickel was greater than 1.0 dm(3), enrichment factor more than 100 and a detection limit of 2.1 microg dm(-3) was achieved. The method applied for detection of nickel in tap water and effluent sample had a relative standard deviation (R.S.D.) of 0.4%. PMID- 19124201 TI - Continuous elimination of Pb2+, Cu2+, Zn2+, H+ and NH4 + from acidic waters by ionic exchange on natural zeolites. AB - A study of breakthrough curves for cations usually found in acid mine drainage (Pb(2+), Cu(2+), Zn(2+) and H(+)) and municipal wastewater (NH(4)(+)) have been conducted on some natural zeolitic tuffs. The zeolitic tuffs used in this study are: three zeolitic tuffs from Cayo Formation, Guayaquil (Ecuador), characterized by X-ray diffraction as clinoptilolite (sample CLI-1) and heulandite (samples HEU 1 and HEU-2)-rich tuffs, and two zeolitic tuffs from Parnaiba Basin, Belem do Para (Brazil), characterized as stilbite-rich tuffs (samples STI-1 and STI-2). The clinoptilolite sample CLI-1 shows an exceedingly good exchange capacities for Pb(2+) and NH(4)(+) as received, and also a very high exchange capacity for Cu(2+) and Zn(2+) when conditioned with 2M sodium chloride, with much higher values than those reported in the literature for other clinioptilolite ores. A general order of effective cation exchange capacity could be inferred from breakthrough curves on these zeolitic tuffs: CLI-1 > HEU-2 > HEU-1 > STI-2. Since it is true for most of the cations studied. PMID- 19124202 TI - Electron beam confinement and image contrast enhancement in near field emission scanning electron microscopy. AB - In conventional scanning electron microscopy (SEM), the lateral resolution is limited by the electron beam diameter impinging on the specimen surface. Near field emission scanning electron microscopy (NFESEM) provides a simple means of overcoming this limit; however, the most suitable field emitter remains to be determined. NFESEM has been used in this work to investigate the W (110) surface with single-crystal tungsten tips of (310), (111), and (100)-orientations. The topographic images generated from both the electron intensity variations and the field emission current indicate higher resolution capabilities with decreasing tip work function than with polycrystalline tungsten tips. The confinement of the electron beam transcends the resolution limitations of the geometrical models, which are determined by the minimum beam width. PMID- 19124203 TI - Immunohistochemical surfactant protein-A expression: fatal drowning vs. postmortem immersion. AB - The postmortem diagnosis of drowning continues to be one of the most difficult in forensic pathology because of unspecific autopsy findings. It must be always remembered that disposal of a victim body in water is not unknown in homicide. The most important physiological consequence in fatal drowning is hypoxemia. The air-liquid interface of alveoli and distal airways of the mature lung are lined with a thin layer of lung surfactant, composed of phospholipids, proteins and neutral lipids. Surfactant components are synthesized and/or incorporated into lung surfactant in alveolar type II cells, and secreted to form an airspace lining film. The composition and function of lung surfactant is disturbed in cases od acute lung injury (ALI) including drowning. Surfactant protein-A (SP-A) is the most abundant surfactant protein. Surfactant protein-A (SP-A) is secreted by type II alveolar cells and cells. It's immunohistochemical distribution is observed in two different pathways: a linear membranous staining and a granular intra-alveolar staining. We hypothesize the significance of immunohistochemical detection of SP-A and its help in determination of the time of death, and possibly distinguishing of death by immersion vs. postmortem immersion using the morphological analysis applied on SP-A immunohistochemical stained lung tissue samples. We also argue in favor of routine use of SP-A staining in selected forensic cases where pathogenesis includes mechanical asphyxia and lung pathology. Although some studies reached conclusions to define the mechanisms involved in pathogenesis of mechanical asphyxia and aspiration necessity of additional studies arose. The mechanism of the production of massive aggregates remains to be determined. In the mean time the detection of SP-A (immunohistochemical) as well as biochemical is potentially useful tool in the forensic practice with possible application in daily practice. PMID- 19124204 TI - Do we need more intensive enzyme replacement therapy for Anderson-Fabry disease? PMID- 19124205 TI - Analysis of APC, alpha-, beta-catenins, and N-cadherin protein expression in aggressive fibromatosis (desmoid tumor). AB - The aims of this study were to analyze the cadherin/catenin adhesion complex in cells from abdominal and extra-abdominal aggressive fibromatosis tumors, and to estimate the correlation between the expression of the tested proteins and the clinical data of the desmoid patients. Immunohistochemistry was used to examine the expression of the cadherin/catenin adhesion complex: APC protein, alpha-, beta-catenin, and N-cadherin in archival material derived from 15 cases of extra abdominal desmoid tumor (E-AD) and 20 cases of abdominal (AD) desmoid tumor. The tested proteins demonstrated cytoplasmic (c) staining. Furthermore, nuclear (n) or cytoplasmic and nuclear (c+n) staining was observed for beta-catenin. The mean values of the percentage of positive cells for the tested proteins between E-AD vs. AD did not demonstrate any statistically significant difference except for alpha-catenin. In the E-AD group, in both cases of recurrent tumors, no alpha catenin expression was observed but the expression of this protein was detected in primary tumors. In the groups investigated, no statistically significant correlation was found between alpha-catenin, beta-catenin (c), (n) and (c+n) expression, and tumor size (p>0.1). The results regarding beta-catenin expression obtained in our study confirm the previous findings that nuclear accumulation of this protein plays a crucial role in the pathogenesis of aggressive fibromatosis. PMID- 19124206 TI - Expression of Toll-like receptor mRNA and cytokines in pigs infected with porcine reproductive and respiratory syndrome virus. AB - Field observations have suggested that porcine reproductive and respiratory syndrome virus (PRRSV) predispose pigs to secondary infections. The interaction between PRRSV and the secondary invaders has not yet been well elucidated. In this study, we investigated the mRNA expression of Toll-like receptors (TLR) in lymphoid organs and cells, and cytokine secretions by alveolar macrophages (AMs) and peripheral blood mononuclear cells (PBMCs) in response to pathogen-associated molecular patterns (PAMPs) in PRRSV-challenged pigs. TLR mRNA expressions were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT PCR) and cytokine concentrations were determined using commercially available ELISA kits. PRRSV infection led to significantly increased secretions of IL-1beta and IL-6 by AMs of PRRSV-infected pigs. Infection of pigs with PRRSV also resulted in an increased secretion of IL-1beta by AMs in response to lipoteichoic acid (LTA) stimulation, and IL-6 by PBMCs in response to lipopolysaccharide (LPS) and LTA stimulation. Infection of pigs with PRRSV tended to up-regulate the mRNA expression of TLR2, 3, 4, 7 and 8 in at least one of the lymphoid tissues and cells. Further research is required to demonstrate the association between the enhanced expressions of the specific TLRs and the increased susceptibility to secondary agents with more severe clinical outcomes in PRRSV-infected pigs. PMID- 19124207 TI - Facial soft tissue thicknesses for the Portuguese adult population. AB - In recent years there has been an important increment of studies on the determination of facial soft tissue thicknesses for craniofacial identification purposes. In spite of this, there is no information on the particularities of the facial soft tissue depths for the Portuguese adult population. In order to address this lack, the soft tissue thicknesses of a Portuguese cadaver sample of 151 individuals, 103 males and 48 females with ages between 20 and 99 years old, autopsied at the South Branch of the National Institute of Legal Medicine, were measured using published needle puncture methods at 20 anatomical points. The stature and weight of all cadavers were determined and their body mass index (BMI) calculated. In order to prevent post-mortem distortion, soft tissue depths were assessed prior to the individual's autopsy in cadavers whose time of death did not exceed 24h. From these data, descriptive statistics were calculated and the average soft tissue depths were reported considering the BMI and the sex of the cadavers. In general, males were found to have greater soft tissue depths than females but some landmarks showed the opposite while others presented the same values for both sexes. In spite of this, almost all the differences among the sexes were found to be not statistically significant. The Post Hoc Gabriel test performed between the different BMI categories for mixed sexes and for each sex suggested BMI-related differences on the soft tissue thicknesses, although these vary considerably within and between the sexes with no linear correspondence between the anatomical landmarks. These results, point not only to the necessity to present data in accordance with the sexual dimorphism, but also to consider the BMI as an important variable for better results in the achievement of the likeness of the deceased individuals in both two-dimensional and three-dimensional facial approximations or superimpositions. Additionally, evidence of interpopulation differences in the average soft tissue thicknesses were observed by the comparison of this data set with similar studies from Caucasian groups with different geographic origin. PMID- 19124208 TI - High-risk human papillomavirus (HPV) is not associated with p53 and bcl-2 expression in oral squamous cell carcinomas. AB - OBJECTIVE: To determine the frequency and type of human papillomavirus (HPV) in oral squamous cell carcinomas (OSCC), as well as to identify a possible association between HPV infection and the expression pattern of p53 and bcl-2, and identify whether the oral HPV infection is a characteristic finding in our sample. METHODS: We performed polymerase chain reaction and dot blot hybridization for the detection of HPV DNA in paraffin sections as well as immunohistochemical analysis of p53 and bcl-2 in our sample. RESULTS: Twenty-six cases (29.5%) were positive for the virus by PCR. Dot blot hybridization identified HPV 18 in 21 (80.8%) cases, HPV 16 in one (3.8%) case and a combination of the two types in the four (15.4%) remaining cases. No other type of HPV was detected in the sample. Immunohistochemistry showed p53 in 26 (60.4%) cases and bcl-2 in 17 (39.5%) ones. No significant association was observed between the presence of HPV and the expression of the proteins studied (p=0.988 and p=0.748, respectively). CONCLUSIONS: Although this investigation have detected only 29.5% of HR-HPV DNA in OSCC, it is possible that this virus contribute to the development of some case of this tumor. Furthermore, it seems that the immunohistochemical expression of p53 and bcl-2 and the presence of HPV DNA are independent events in OSCC. PMID- 19124209 TI - Efficacy of a long-term antibiotic treatment in patients with a chronic Tick Associated Poly-organic Syndrome (TAPOS). AB - SETTINGS: Despite a now codified antibiotic treatment for Lyme disease, a significant proportion of patients treated according to recommendations complain of persistent signs and symptoms. The pathophysiological mechanisms which underlie this syndrome of post-treatment chronic systemic illness remain unclear. For some physicians post-treatment symptoms indicate a persistent infection requiring prolonged antibiotic therapy. For others, there is no benefit from antimicrobial therapy. The difficulty of assessment encountered in studies is significant because many symptoms are subjective. We think that the term "chronic Lyme disease" is not appropriate and should be replaced by chronic "tick associated poly-organic syndrome" (TAPOS). OBJECTIVE: This open-label prospective study was made on a group of 100 patients having followed a medical treatment for a chronic TAPOS and to evaluate their evolution under prolonged antibiotic treatment. RESULTS: The medical management was found to be effective for symptoms, especially for patients with a high probability of chronic TAPOS (NEJM score). Patients with post tick-bite symptoms, which often worsens their quality of life, deserve particular attention. CONCLUSION: This study had methodological limitations but could help in terms of feasibility, choice of inclusion criteria, and design of follow-up for a future randomized, double blind study to test for an optimal management of TAPOS. PMID- 19124210 TI - [Cholangiocarcinoma and humoral hypercalcemia of malignancy]. PMID- 19124212 TI - Shear and extensional rheology of hydroxypropyl cellulose melt using capillary rheometry. AB - With increasing interest in hot melt extrusion for preparing polymer-drug systems, knowledge of the shear and extensional rheology of polymers is required for the formulation and process design. Shear and extensional rheology of three commercial grades of hydroxypropyl cellulose (HPC) was examined at 140, 145 and 150 degrees C using twin bore capillary rheometry at range of processing rates. The power law model fitted for shear flow behaviour up to shear strain rates of approximately 1000s(-1), above which measured shear viscosities deviated from the power law and surface instabilities were observed in the extrudate, particularly for higher molecular weight grades. Shear thinning index was found to be relatively independent of temperature and molecular weight, whilst the consistency index, indicative of zero shear viscosity increased exponentially with increase in molecular weight. Extensional viscosity of all grades studied was found to decrease with increasing temperature and increasing processing rate. Foaming of the extrudate occurred especially at low temperatures and with the high molecular weight grade. An understanding of the relationships between shear and extensional flows with temperature, processing rate and molecular weight is a useful tool for process design; optimisation and troubleshooting of Hot melt extrusion (HME) of pharmaceutical formulations. PMID- 19124213 TI - Development and validation of HPLC method for the determination of Cyclosporin A and its impurities in Neoral capsules and its generic versions. AB - Cyclosporin A (CyA) is a cornerstone immunosuppressant for the prophylaxis against allograft rejection after organ transplantation. The most widely prescribed CyA formulation is Neoral soft gelatine capsules (Novartis Pharmaceuticals, Basel, Switzerland). After Novartis patent expiration, several generic formulations have been developed. In this paper, a simple and reliable HPLC method was developed and validated for the evaluation of four CyA degradation products (ID-005-95, CyH, IsoCyH and IsoCyA) and two related compounds (CyB and CyG) aimed for the quality control of Neoral capsules and its generic formulations. In a second step, the validated method was then compared to the USP assay method for capsules, where some of the mentioned impurities were not adequately resolved from the CyA peak. Isocratic elution at a flow rate of 1.0mLmin(-1) was employed on a Lichrospher RP-18 (4mmx250mm; 5microm) analytical column maintained at 75 degrees C with a tetrahydrofuran:phosphoric acid (0.05M) (44:56, v/v) as mobile phase. The chromatograms were recorded using a Hewlett Packard 1100 chromatographic system. The UV detection wavelength was performed at 220nm and 10microL of sample was injected. The developed method was validated in terms of selectivity, linearity, precision, accuracy, limit of detection and limit of quantitation. The validate method was successfully applied to commercial capsules, Neoral and generic versions. Therefore, the proposed method is suitable for the simultaneous determination of CyA as well as its major impurities. PMID- 19124214 TI - Development and validation of an HPLC-DAD method for bis(12)-hupyridone and its application to a pharmacokinetic study. AB - A rapid and simple method of high performance liquid chromatography with UV detection for the quantification of bis(12)-hupyridone in rat blood has been developed and validated. Chromatographic separation was carried out in an Agilent Extend C(18) 5microm column (length, 250mm; inner diameter, 4.6mm) using a mixture of water-acetonitrile-trifluoroacetic acid (81:19:0.04, v/v/v) as the mobile phase at a flow rate of 1mL/min, with detection at 229nm. The method used for the bis(12)-hupyridone quantification showed linearity for concentration range of 0.1-7.5microg/mL with r(2)=0.9991. The limit of detection and quantification of this method were 0.05microg/mL and 0.1microg/mL, respectively. The intra- and inter-day variations of the analysis were less than 4.22% with standard errors less than 13.3%. The developed method was successfully applied to the pharmacokinetic study of bis(12)-hupyridone after intravenous administration of 5mg/kg and intraperitoneal administration of 10 and 20mg/kg in rats. PMID- 19124215 TI - Increasing mental health awareness and appropriate service use in older Chinese Americans: a pilot intervention. AB - OBJECTIVE: This study evaluated the effectiveness of a community intervention in increasing awareness of mental health issues and available resources among elderly Chinese Americans. METHODS: Twenty-seven members of a community church received a 1-h didactic presentation, in English and Mandarin, and completed surveys regarding their help-seeking preferences before and after the intervention. Results were analyzed using a series of Wilcoxon matched-pair signed rank tests and comparing pre- and post-test scores. RESULTS: Findings indicated an increase (p<.05) in intention to consult a mental health professional for psychiatric symptoms at post-test. A significant increase also was found in preference for consulting a physician for physical symptoms. CONCLUSION: The pilot educational intervention increased awareness of mental health and treatment issues and the role of mental health professionals, lending support to evaluate the intervention on a larger scale. PRACTICE IMPLICATIONS: Greater awareness of mental health among Chinese Americans can be promoted via education forums provided through faith-based organizations. Stigma of mental illness leads many Chinese individuals to seek help for psychiatric problems from primary care physicians. Integrating mental health practitioners in primary care settings may help decrease stigma and encourage appropriate help-seeking behavior. PMID- 19124216 TI - Analyzing the "nature" and "specific effectiveness" of clinical empathy: a theoretical overview and contribution towards a theory-based research agenda. AB - OBJECTIVE: To establish sound empirical evidence that clinical empathy (abbreviated as CE) is a core element in the clinician-patient relationship with profound therapeutic potential, a substantial theoretical-based understanding of CE in medical care and medical education is still required. The two aims of the present paper are, therefore, (1) to give a multidisciplinary overview of the "nature" and "specific effectiveness" of CE, and (2) to use this base as a means of deriving relevant questions for a theory-based research agenda. METHOD: We made an effort to identify current and past literature about conceptual and empirical work focusing on empathy and CE, which derives from a multiplicity of disciplines. We review the material in a structured fashion. RESULTS: We describe the "nature" of empathy by briefly summarizing concepts and models from sociology, psychology, social psychology, education, (social-)epidemiology, and neurosciences. To explain the "specific effectiveness" of CE for patients, we develop the "Effect model of empathic communication in the clinical encounter", which demonstrates how an empathically communicating clinician can achieve improved patient outcomes. Both parts of theoretical findings are synthesized in a theory-based research agenda with the following key hypotheses: (1) CE is a determinant of quality in medical care, (2) clinicians biographical experiences influence their empathic behavior, and (3) CE is affected by situational factors. CONCLUSION: The main conclusions of our review are twofold. First of all, CE seems to be a fundamental determinant of quality in medical care, because it enables the clinician to fulfill key medical tasks more accurately, thereby achieving enhanced patient health outcomes. Second, the integration of biographical experiences and situational factors as determinants of CE in medical care and medical education appears to be crucial to develop and promote CE and ultimately ensuring high-quality patient care. PRACTICE IMPLICATIONS: Due to the complexity and multidimensionality of CE, evidence-based investigations of the derived hypotheses require both well-designed qualitative and quantitative studies as well as an interdisciplinary research approach. PMID- 19124217 TI - Isolated tumour cells versus micrometastases and non-sentinel node involvement in breast cancer. PMID- 19124218 TI - Endovascular treatment of renal artery aneurysm secondary to fibromuscular dysplasia: a case report. AB - We present a case where a saccular aneurysm of the left renal artery in a patient with fibromuscular dysplasia (FMD) was successfully treated by an endovascular approach utilizing two covered 6 x 22 mm stent grafts. Aneurysm formation is a known complication of FMD. There are multiple treatment options for dealing with these aneurysms, both open and endovascular. This case report describes management of a left renal artery aneurysm with covered stents. PMID- 19124219 TI - Conference abstracts of a new oncology drug do not always lead to full publication: proceed with caution. AB - BACKGROUND: Conference abstracts, often the first public record of a study, serve as a catalyst to initiate clinical and policy change. On average, 45% of all conference abstracts subsequently appear as full publications; however, the generalizability of this finding to studies of one intervention, in one population, is unknown. Our objectives were to determine the full publication rate of a cohort of abstracts, median time to publication, and predictors of these relationships. METHODS: We included the first 5 years of clinical abstract reports of rituximab for non-Hodgkin's lymphoma (NHL) from American Society of Hematology (ASH) meetings (1997-2001), identified all unique studies, and used electronic databases to identify full publications. We determined the full publication rate, median time to publication, and predictors of these outcomes. RESULTS: Of 109 abstracts representing 86 unique studies, the publication rate was 52.3% (45, 95% confidence interval [CI]: 41.3, 63.2), and the median time to publication, 1.4 years with 6.8 years' follow-up. Author affiliation with industry (odds ratio [OR] [95% CI]=4.60 [1.32, 16.08] and presentation type (oral OR=5.94 [1.31, 26.88], poster OR=3.39 [1.24, 9.25]; reference, publication in conference abstract book only) independently predicted subsequent full publication in the adjusted analysis. We identified no predictors of time to publication. INTERPRETATION: We suggest cautious consideration of data from conference proceedings to inform new technology clinical or policy decisions. Future work needs to examine the generalizability of our results to other diseases and technologies. PMID- 19124220 TI - Mathematical weighting of a clinimetric index (Pediatric Ulcerative Colitis Activity Index) was superior to the judgmental approach. AB - BACKGROUND: We aimed to compare the judgmental and mathematical approaches in weighting the Pediatric Ulcerative Colitis Activity Index (PUCAI). METHODS: The PUCAI was previously weighted mathematically using multivariate regression modeling on 157 children with ulcerative colitis (UC). Independently, a Delphi group of 36 experts in pediatric UC judgmentally provided weights to the PUCAI's items. The agreement between the tools was evaluated using the 95% limits of agreement method. Validity was assessed on a prospective cohort of 48 UC children, using three constructs: colonoscopic appearance, physician's global assessment, and the Mayo score. Responsiveness was compared on a longitudinal cohort of 75 children. RESULTS: The weights of the resulting PUCAI tools were quite similar, but the Delphi group retained the laboratory items, excluded by the mathematical modeling. This difference was reflected by the Bland and Altman method. Both tools performed equally well in the validation and responsiveness cohorts. CONCLUSION: The judgmentally weighted PUCAI had good validity and responsiveness. The mathematical weighting, however, performed just as well without the laboratory items, resulting in a more feasible index. Therefore, the mathematical modeling has proven to be superior in weighting the PUCAI. PMID- 19124221 TI - New strategies are needed to improve the accuracy of influenza vaccine effectiveness estimates among seniors. AB - OBJECTIVE: The magnitude of the benefit of influenza vaccine among elderly individuals has been recently debated. Existing vaccine effectiveness estimates derive primarily from observational studies, which may be biased. In this paper, we provide a methodological examination of the potential sources of bias in observational studies of influenza vaccine effectiveness in seniors and propose design and analysis strategies to reduce bias in future studies. STUDY DESIGN AND SETTING: We draw parallels to bias documented in observational studies of therapies in other areas of medical research including pharmacoepidemiology, discuss reasons why existing adjustment methods in influenza studies may not adequately control for the bias, and evaluate statistical approaches that may yield more accurate estimation of influenza vaccine effectiveness. RESULTS: There is strong evidence for the presence of bias in existing observational estimates of influenza vaccine effectiveness in the elderly and the failure of current adjustment methods to reduce bias. CONCLUSION: Promising approaches for reducing bias include obtaining more accurate information on confounders, such as functional status, avoiding all-cause death in favor of outcomes, such as pneumonia or influenza-related pneumonia, and evaluating the extent to which bias is reduced by these and other methods using the 'control' period before influenza season. PMID- 19124222 TI - Inactivated influenza vaccines: methods, policies, and politics. PMID- 19124224 TI - New approach to gallbladder ultrasonic images analysis and lesions recognition. AB - This paper presents a new approach to gallbladder ultrasonic image processing and analysis towards detection of disease symptoms on processed images. First, in this paper, there is presented a new method of filtering gallbladder contours from USG images. A major stage in this filtration is to segment and section off areas occupied by the said organ. In most cases this procedure is based on filtration that plays a key role in the process of diagnosing pathological changes. Unfortunately ultrasound images present among the most troublesome methods of analysis owing to the echogenic inconsistency of structures under observation. This paper provides for an inventive algorithm for the holistic extraction of gallbladder image contours. The algorithm is based on rank filtration, as well as on the analysis of histogram sections on tested organs. The second part concerns detecting lesion symptoms of the gallbladder. Automating a process of diagnosis always comes down to developing algorithms used to analyze the object of such diagnosis and verify the occurrence of symptoms related to given affection. Usually the final stage is to make a diagnosis based on the detected symptoms. This last stage can be carried out through either dedicated expert systems or more classic pattern analysis approach like using rules to determine illness basing on detected symptoms. This paper discusses the pattern analysis algorithms for gallbladder image interpretation towards classification of the most frequent illness symptoms of this organ. PMID- 19124223 TI - Response analysis for multiple symptoms revealed differences between arms of a symptom management trial. AB - OBJECTIVE: To describe the methodology of evaluating the response of cancer patients to interventions directed at lowering severity of multiple symptoms, and to compare two arms of a symptom management trial to determine factors associated with response and time to response. STUDY DESIGN AND SETTING: Randomized trial comparing a nurse-assisted symptom management (NASM) cognitive-behavioral intervention with an automated telephone symptom management (ATSM). Patients in both arms received six intervention contacts over 8 weeks. Analyses of the intervention contact data for 190 patients in NASM arm and 164 patients in the ATSM arm were conducted. Severities of 15 cancer-related symptoms were assessed at each intervention contact, and an anchor-based definition of response was adopted. Analyses were carried out using generalized estimating equations and Cox marginal proportional hazard models. RESULTS: When compared with patients in the NASM, patients in the ATSM had better response to manage anxiety, depression, poor appetite, cough, and fatigue. NASM was more successful in managing cancer pain. Response and time to response were associated with several patient and disease characteristics. CONCLUSION: The approach described here presents an analytic and clinical improvement over methods that examine each symptom separately or use summed scores of severity. PMID- 19124226 TI - Long-term outcome of childhood absence epilepsy: Dutch Study of Epilepsy in Childhood. AB - SUMMARY: We determined long-term outcome and the predictive value of baseline and EEG characteristics on seizure activity evolution in 47 children with newly diagnosed childhood absence epilepsy (CAE) included in the Dutch Study of Epilepsy in Childhood. All children were followed for 12-17 years. The children were subdivided in three groups for the analyses: those becoming seizure-free (I) within 1 month after enrolment; (II) 1-6 months after enrolment; and (III) more than 6 months after enrolment or having seizures continuing during follow-up. No significant differences were observed between groups in sex, age at onset, occurrence of febrile seizures, and positive first-degree family history for epilepsy. All groups had high remission rates after 12-17 years. Significantly more relapses occurred in group III than in group I. Total duration of epilepsy and mean age at final remission were 3.9 and 9.5 years, respectively, being significantly longer and higher in group III than in groups I and II. In all groups only a small number of children (total 13%) developed generalized tonic clonic seizures. In conclusion, our children with CAE had an overall good prognosis with few children (7%) still having seizures after 12-17 years. Remission rate in children with CAE cannot be predicted on the basis of baseline and EEG characteristics. The early clinical course (i.e. the first 6 months) has some predictive value with respect to the total duration of absence epilepsy. PMID- 19124227 TI - Tactile impression and friction of water on human skin. AB - Water has a unique touch as well as characteristic physical properties. However, nobody knows the real identity of its touch. Here, we show that water creates a stick-slip feel when a small amount is rubbed using fingertip on an artificial skin that mimics the structure of human skin. The results of frictional analyses predict that this stick-slip feel is caused by a drastic change in frictional resistance. The present result is valuable for biologists and robot engineers as well as cognitive scientists and tribologists, because it is a new example of stick-slip phenomena on biological surfaces. The tactile texture of this most familiar material could also be applied to consumer products or virtual reality systems. PMID- 19124225 TI - Zonisamide block of cloned human T-type voltage-gated calcium channels. AB - SUMMARY: Zonisamide (ZNS) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures, with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action. In this study, we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous HEK-293 expression system using whole cell patch-clamp technique. Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8% reduction of Ca(2+) influx within the maximum therapeutic plasma range (50-200 microM ZNS). The other T-type Ca(2+) channel entities, Ca(v)3.1 and Ca(v)3.3, were even less sensitive to ZNS. Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC, whereas Ca(v)3.1 and Ca(v)3.3 exhibited minor, though significant reduction of inactivation-tau. Interestingly, ZNS block of Ca(v)3.2 VGCCs was not use dependent and remained unaffected by changes in the holding potential. Steady state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS (DeltaV(1/2)=3.1mV, p=0.071). Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics, use- and state-dependence of blockade. These results suggest that T type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug. PMID- 19124228 TI - Self-aggregated nanoparticles from linoleic acid modified carboxymethyl chitosan: Synthesis, characterization and application in vitro. AB - The purpose of the present research work was to study the formation of linoleic acid (LA) modified carboxymethyl chitosan (LCC). Another objective was to evaluate effect of linoleic acid degree of substitution on loading capacity (LC), ADR loading efficiency (LE) and in vitro release profile of LCC nanoparticles. The hydrogel nanoparticles can be prepared using linoleic acid modified carboxymethyl chitosan (LACMCS) after the sonication. The critical aggregation concentration (CAC) of the self-aggregate of LA modified CMCS (LCC) was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC values were in the range of 0.061-0.081mg/mL. Self-aggregated nanoparticles exhibited an increased LC and LE, decreased sustained release with an increasing ratio of the hydrophobic LA to hydrophilic CMCS. LCC nanoparticles loaded with ADR exerted in vitro anticancer activity against Hela cells that was comparable to the activity of free (non-entrapped in nanoparticles) ADR. PMID- 19124229 TI - Behavior of sulfatide/cholesterol mixed monolayers at the air/water interface. AB - The monolayer properties of sulfatide and cholesterol binary system have been investigated with surface pressure-mean molecular area isotherms measurements and atomic force microscopy (AFM). The thermodynamic analysis indicates that the obtained negative deviation of the excess molecular area (DeltaA((ex))) and surface excess Gibbs energy (DeltaG((ex))) from the ideal behavior at various molar ratios, suggesting an attractive interaction between sulfatide and cholesterol in the monolayers as compared with the pure components monolayers. Meanwhile, the compression modulus (C(s)(-1)) vs. surface pressure (pi) and activity coefficients (f(1) and f(2)) of mixed films dependencies for mixed monolayers are drawn at different mole fractions. The AFM images for the mixed sulfatide/cholesterol monolayers deposited on the mica at 15 and 30mNm(-1) show the stronger molecular attractive force to form condensed structure. The behavior of sulfatide is thought to be involved in lateral domain formation in biological membranes. Therefore, the interaction between sulfatide and cholesterol becomes more important in mimicking "lipid rafts" domains. PMID- 19124230 TI - IDM release behavior and surface characteristics of the novel Cu/IDM/LDPE nanocomposite for intrauterine device. AB - Copper/indomethacin/low-density polyethylene (Cu/IDM/LDPE) nanocomposite was prepared as a novel material for intra-uterine device (IUD). IDM release profile of the nanocomposite was investigated by using spectrophotometer. The results show that IDM release rate of Cu/IDM/LDPE nanocomposite is higher in simulated uterine solution than that in methanol, confirming that the release process of IDM is dominated mainly by pore diffusion. The decrease in copper particle size and the increase in copper mass content all accelerate IDM release, indicating that IDM release rate can be adjusted by changing copper loading or copper particle size. The surface of the incubated nanocomposite was characterized by X ray diffraction, scanning electron microscopy and energy dispersive X-ray microanalysis. A few deposits composed of P, Cl, Ca, Cu and O were observed on the nanocomposite surface, which may be related to the presence of IDM particles with large particle size. PMID- 19124231 TI - Preparation and drug controlled-release of polyion complex micelles as drug delivery systems. AB - Block copolymers, poly(N-vinylprrolidone)-block-poly(styrene-alter-maleic anhydride) (PVP-b-PSMA) and poly(N-vinylprrolidone)-block-poly(N,N dimethylaminoethyl methacrylate) (PVP-b-PDMAEMA), were synthesized by reversible addition- fragmentation chain transfer (RAFT) polymerization. In aqueous media, this a pair of oppositely-charged diblock copolymers could self-assemble into stable and narrow distribution polyion complex micelles (PICMs). Transmission electron micrographs (TEM) and dynamic light scattering (DLS) analysis showed that the micelles to be spherically shaped with mean hydrodynamic diameter around 70nm. In addition, the PICMs display ability to response to external stimuli. All of theses features are quite feasible for utilizing it as a novel intelligent drug delivery system. In order to assess its application in biomedical area, release profiles of coenzyme A (Co A) from PICMs were studied under both simulated gastric and intestinal pH conditions. The release was much quicker in pH 7.4 buffer than in pH 2.0 solution. Based on these results, these PICMs could be a potential pH-sensitive carrier for colon-specific drug delivery system. PMID- 19124232 TI - The pterygopalatine ganglion in humans: a morphological study. AB - As a rule the pterygopalatine ganglion (PPG) is considered to be a single structure of the parasympathetic nervous system, associated with the maxillary nerve in the pterygopalatine fossa (PPF). A few structural studies in humans are available in the indexed references. We designed the present study of the PPG in order to provide evidence of possible variations in morphological patterns of the PPG. We performed dissections of the PPF on 20 human adult heads, using different approaches. The dissected specimens were stained with hematoxylin-eosin and silver (Bielschowsky) or prepared for immunohistochemistry for synaptophisin and neurofilament. Four morphological types of the PPG were defined macroscopically: A (10%): partitioned PPG, the upper partition receiving the vidian nerve; B (55%): single, the upper part (base) receiving the vidian nerve; C (15%): single, but the vidian nerve reaches the lower part (tip) of the ganglion; D (20%): partitioned, the lower partition receiving the vidian nerve. We propose that it may be inappropriate to invariably regard the PPG as a single morphological structure. From individual to individual the PPG may present either as a single ganglion or as a partitioned one, with distinct superior and inferior components. Nevertheless, the presence of the dispersed pterygopalatine microganglia (DPPG) evidenced by histochemistry and immunohistochemistry serves to complete an individually variable morphological pattern of a structure usually described as single. The individual variation may be the reason for failures in ablation procedures of the PPG; partitions of the PPG and/or the DPPG may functionally correlate with specific territories and targets and further tracing studies may be helpful in validating or invalidating this theory. PMID- 19124233 TI - Moral regulation and the presumption of guilt in Health Canada's medical cannabis policy and practice. AB - This paper is a sociological examination of policies and practices in Health Canada's Marihuana Medical Access Division (MMAD) that presume the illicit intentions and inherent "guilt" of medical cannabis users, hampering safe access to a medicine to which many are legally entitled, and raising doubts about this federal programme's overall effectiveness and constitutional legitimacy. Beginning with a brief historical overview of Canada's federal medical cannabis programme, this paper examines the failure of the MMAD to meet the needs of many sick and suffering Canadians through Hunt's [Hunt, A. (1999). Governing morals: A social history of moral regulation. Cambridge, UK: Cambridge University Press] work on moral regulation and Wodak's [Wodak, A. (2007). Ethics and drug policy. Psychiatry, 6(2), 59-62] critique of "deontological" drug policy strategies. I then cite Tupper's [Tupper, K. W. (2007). The globalization of ayahuasca: Harm reduction or benefit maximization? International Journal of Drug Policy, doi:10.1016/j.drugpo.2006.11.001] argument that shifting to a generative metaphor that constructs certain entheogenic substances as potentially useful "tools" rather than regulating them through inherently moralistic prohibitionist policies would better serve public health, and incorporate Young's [Young, I. M. (1990). Justice and the politics of difference. Princeton, New Jersey: Princeton University Press] theories of domination and oppression to examine the rise of community-base medical cannabis dispensaries as "new social movements". First hand accounts by medical cannabis patients, federally funded studies, and internal Health Canada communication and documents suggest that current federal policies and practices are blocking safe access to this herbal medicine. The community-based dispensary model of medical cannabis access is a patient-centered "new social movement" that mitigates the stigmatization and moral regulation of their member-clients by creating opportunities for engagement, empowerment and joint knowledge creation. In light of ongoing Charter challenges and patient criticism, the survival of this federal programme will depend on the government's ability to shift away from policies based on the oppression and moral regulation, and towards consequentialist policies that balance harm reduction and benefit maximization. The effectiveness of such an approach is exemplified by the success of the community-based dispensary model which is currently producing more peer reviewed research and supplying medical cannabis to a far greater number of patients than Health Canada's Marihuana Medical Access Division. PMID- 19124234 TI - Characterization of ceramic roof tile wastes as pozzolanic admixture. AB - The aim of this work is to study the recycling of tile wastes in the manufacture of blended cements. Cracked or broken ceramic bodies are not accepted as commercial products and, therefore, the unsold waste of the ceramic industry becomes an environment problem. The use of powdered roof tile in cement production, as pozzolanic addition, is reported. The wastes were classified as nonglazed, natural and black glazed tiles. The mineralogy of the powders was controlled by SEM-EDX microscopy, XRD analysis and FTIR spectroscopy. Particle size was checked by laser granulometry. Once the materials were fully characterized, pozzolanic lime consumption tests and Fratini tests were carried out. Different formulations of cement-tile blends were prepared by incorporation of up to 30% weight ratios of recycled waste. The compressive strength of the resulting specimens was measured. The evolution of hydration of the cement-tile blends was analyzed by XRD and FTIR techniques. Vibrational spectroscopy presented accurate evidence of pozzolanic activity. The results of the investigation confirmed the potential use of these waste materials to produce pozzolanic cement. PMID- 19124235 TI - A simple strategy of probe DNA immobilization by diazotization-coupling on self assembled 4-aminothiophenol for DNA electrochemical biosensor. AB - A novel and simple strategy for fabricating of DNA electrochemical biosensor was developed based on covalent coupling of probe NH(2)-ssDNA (S(1)) on Au electrode that had been functionalized by diazotization of assembled 4-aminothiophenol (4 ATP) monolayer. The thiol group of 4-ATP allowed the stable assembly of 4-ATP monolayer. The following diazotization reaction was directly performed to prepare functional diazo-ATP film for covalent coupling of probe S(1). Remarkably, it was noting that the diazo-ATP provided a surface with high conductibility for electron transfer. The complementary ssDNA was determined by using differential pulse voltammetry. The linear range of the developed biosensor was from 1.57 x 10(-9) to 4.52 x 10(-7)M with a detection limit of 3.26 x 10(-10)M. The fabricated biosensor possessed good selectivity and could be regenerated. The covalent immobilization of probe S(1) by simple diazotization-coupling on self assembled 4-ATP monolayer could serve as a versatile platform for DNA immobilization and biosensors fabricating. PMID- 19124236 TI - Development of an electrochemical immunoassay for rapid detection of E. coli using anodic stripping voltammetry based on Cu@Au nanoparticles as antibody labels. AB - A sensitive electrochemical immunoassay for rapid detection of Escherichia coli has been developed by anodic stripping voltammetry (ASV) based on core-shell Cu@Au nanoparticles (NPs) as anti-E. coli antibody labels. The characteristics of Cu@Au NPs before and after binding with antibody were confirmed by transmission electron microscopy (TEM). After Cu@Au-labeled antibody reacted with the immobilized E. coli on Polystyrene (PS)-modified ITO chip, Cu@Au NPs were dissolved by oxidation to the metal ionic forms, and the released Cu(2+) ions were determined at GC/Nafion/Hg modified electrode by ASV. The utilization of GC/Nafion/Hg modified electrode could enhance the sensitivity for Cu(2+) detection with a concentration as low as 9.0 x 10(-12)mol/L. Since Cu@Au NPs labels were only present when antibody reacted with E. coli, the amount of Cu(2+) directly reflected the number of E. coli. The technique could detect E. coli with a detection limit of 30CFU/mL and the overall analysis could be completed in 2h. By introducing a pre-enrichment step, a concentration of 3CFU/10mL E. coli in surface water was detected by the electrochemical immunoassay. PMID- 19124237 TI - Resistance to EGF-R (erbB-1) and VEGF-R modulating agents. AB - In an effort to improve the survival of cancer patients, new therapeutic approaches focusing on the molecular mechanisms that mediate tumour cell growth or survival have gained much attention. In particular, EGF-R and VEGF/VEGF-R have been extensively investigated as targets for anti-neoplastic therapy. Agents that selectively target EGF-R, erbB-2, VEGF-R-2 or VEGF have shown promising activity in clinical trials, and several are now approved for use in selected cancer indications. However, all patients ultimately develop resistance to these drugs. Thus, there is a great need to understand how patients become resistant to effective therapies for these cancers since this approach may lead to improvements in therapies that target EGF-R and VEGF/VEGF-R. Pre-clinical studies have begun to shed light on the mechanisms of resistance to anti-angiogenetic drugs and to date four mechanisms of resistance have been identified (1) upregulation of bFGF, (2) overexpression of MMP-9, (3) increased levels of SDF 1alpha and (4) HIF-1alpha-induced recruitment of bone marrow-derived CD45+ myeloid cells. In addition, the molecular mechanisms of resistance to EGF-R modulating agents can be attributed to several general processes: (1) activation of alternative tyrosine kinase inhibitors that bypass the EGF-R pathway (e.g. c MET and IGF-1R), (2) increased angiogenesis, (3) constitutive activation of downstream mediators (e.g. PTEN and K-ras) and (4) the existence of specific EGF R mutations. K-ras mutations have been significantly associated with a lack of response to EGF-R tyrosine kinase inhibitors in patients with NSCLC and with a lack of response to cetuximab or to panitumumab in patients with advanced colorectal cancer. The identification of these resistance mechanisms has led to clinical trials using newly designed targeted therapies that can overcome resistance and have shown promise in laboratory studies. Ongoing research efforts will likely continue to identify additional resistance mechanisms, and these findings will hopefully translate into effective therapies for different cancers. PMID- 19124238 TI - Endobronchial ultrasound-guided transbronchial needle aspiration for staging of lung cancer: a systematic review and meta-analysis. AB - STUDY OBJECTIVES: Recently, less invasive methods have emerged as potential alternatives for staging with tissue confirmation of suspected metastatic mediastinal lymph nodes in lung cancer. The objective of this review was to assess the overall diagnostic accuracy of EBUS-TBNA in detecting metastatic mediastinal lymph node in lung cancer with a meta-analysis. METHODS: The MEDLINE, EMBASE, Cancerlit and Cochrane Library database, from January 1995 to September 2008, were searched for studies evaluating EBUS-TBNA accuracy. Meta-analysis methods were used to pool sensitivity and specificity and to construct summary receiver-operating characteristic. RESULTS: A total of 11 studies with 1299 patients, who fulfilled all of the inclusion criteria, were considered for the analysis. No publication bias was found. EBUS-TBNA had a pooled sensitivity of 0.93 (95% CI, 0.91-0.94) and a pooled specificity of 1.00 (95% CI, 0.99-1.00). The subgroup of patients who were selected on the basis of CT or PET positive results had higher pooled sensitivity (0.94, 95% CI 0.93-0.96) than the subgroup of patients without any selection of CT or PET (0.76, 95% CI 0.65-0.85) (p<0.05). Study sensitivity was not correlated with the prevalence of lymph node metastasis. Only two complications occurred (0.15%). CONCLUSION: EBUS-TBNA was an accurate, safe and cost-effective tool in lung cancer staging. The selection of patients who had positive results of suspected lymph node metastasis in CT or PET may improve the sensitivity of EBUS-TBNA. High-quality prospective studies regarding EBUS-TBNA in lung cancer staging are still needed to be conducted. PMID- 19124240 TI - Effect of the oligo(ethylene glycol) group on the antioxidant activity of manganese salen complexes. AB - The synthesis and antioxidant activity of oligo(ethylene glycol)-modified manganese salen complexes are reported. Their SOD activities were similar and 2- to 3-fold more potent than the standard compound EUK-134. Their catalase-like activity was lower than that of EUK-134 in the initial conversion rate; however, some analogs exhibited a better catalytic turnover number. PMID- 19124241 TI - Reactivation potency of fluorinated pyridinium oximes for acetylcholinesterases inhibited by paraoxon organophosphorus agent. AB - For the purpose of developing new oxime reactivators of acetylcholinesterases (AChE) that have been inhibited by organophosphorus agents, emphasis was given to the finding that the lipophilic nature of fluorinated compounds is responsible for their enhanced transport across the blood brain barrier (BBB). As a result, we have designed and synthesized the fluorinated oxime derivatives, which quantum mechanical calculations suggest should have a greater lipophilicity and BBB permeability than their non-fluorinated analogs. Among the compounds explored in this study, 4 was found to have the highest potency for reactivation of paraoxon inhibited housefly (HF) AChE. PMID- 19124242 TI - Design and synthesis of amidoxime derivatives for orally potent C-alkylamidine based antimalarial agents. AB - Within the frame of the design of prodrug candidates to deliver a C-alkylamidine antimalarial agent, we showed that specific O-substitutions were needed on the alkylamidoxime structure. Among the newly synthesized molecules, bis-oxadiazolone and bis-O-methylsulfonylamidoxime derivatives induced a complete clearance of parasitemia in mice after oral administration. PMID- 19124243 TI - 3,4-Disubstituted isothiazoles: novel potent inhibitors of VEGF receptors 1 and 2. AB - Novel derivatives of isothiazoles are described as potent ATP-competitive inhibitors of vascular endothelial growth factor receptors I and II (VEGFR-1/2). A number of compounds exhibited VEGFR-2 inhibitory activity comparable to that of Vatalanib in both HTRF enzymatic and cellular assays. Several derivatives featuring bulky meta-substituents in the amide portion of the molecule displayed 4- to 8-fold specificity for VEGFR-2 versus VEGFR-1. Active molecules also showed high intrinsic permeability (> 30 x 10(-5) cm/min) across Caco-2 cell monolayer. PMID- 19124239 TI - Survival trends in European cancer patients diagnosed from 1988 to 1999. AB - We analysed data from 49 cancer registries in 18 European countries over the period 1988-1999 to delineate time trends in cancer survival. Survival increased in Europe over the study period for all cancer sites that were considered. There were major survival increases in 5 year age-adjusted relative survival for prostate (from 58% to 79%), colon and rectum (from 48% to 54% men and women), and breast (from 74% to 83%). Improvements were also significant for stomach (from 22% to 24%), male larynx (from 62% to 64%), skin melanoma (from 78% to 83%), Hodgkin disease (from 77% to 83%), non-Hodgkin lymphoma (from 49% to 56%), leukaemias (from 37% to 42%), and for all cancers combined (from 34% to 39% in men, and from 52% to 59% in women). Survival did not change significantly for female larynx, lung, cervix or ovary. The largest increases in survival typically occurred in countries with the lowest survival, and contributed to the overall reduction of survival disparities across Europe over the study period. Differences in the extent of PSA testing and mammographic screening, and increasing use of colonoscopy and faecal blood testing together with improving cancer care are probably the major underlying reasons for the improvements in survival for cancers of prostate, breast, colon and rectum. The marked survival improvements in countries with poor survival may indicate that these countries have made efforts to adopt the new diagnostic procedures and the standardised therapeutic protocols in use in more affluent countries. PMID- 19124244 TI - Osteogenic activity of constituents from Butea monosperma. AB - Phytochemical investigation from the stem bark of Butea monosperma, led to the isolation and identification of three new compounds named buteaspermin A (1), buteaspermin B (2) and buteaspermanol (3), along with 19 known compounds. The structure of compounds 1-22 were established on the basis of their spectroscopic data. The isolated compounds 2-17 were evaluated using neonatal (1-3 day old) rat calvaria derived primary osteoblast cultures. Five of these compounds 7, 10-13 showed promising osteogenic activity, attributed to increased osteoblast proliferation, differentiation and mineralization as evidenced by marked increase in expression of alkaline phosphatase, an early phase differentiation marker, and alizarin Red S staining of osteoblasts cultured for 48 h and von Kossa silver staining of nodules formed 15 days after culture with these compounds. Quantification of mineralization by optical density measurement of Alizarin Red S extracted from stained osteoblasts cultured for 7 days in presence of these compounds showed significant (P<0.05, vs corresponding vehicle control group) increase in mineralization. On the basis of biological results, structure activity relationships are discussed. PMID- 19124245 TI - Changes in the coordination of hip and pelvis kinematics with mode of locomotion. AB - Limited hip extension mobility has been proposed as a possible cause of both increased anterior pelvic tilt and subsequent exaggerated lumbar lordosis during walking and running. The purpose of the present study was to examine the coordinated sagittal plane kinematic patterns of the hip and pelvis during walking and running in a substantial group of adult recreational runners. The kinematics of 73 healthy adult runners (age: 34+/-11years) were examined on an instrumented treadmill at self-selected walking and running speeds using a three dimensional motion capture system. Although stride length increased considerably from walking to running, the range of hip extension utilized during running was not significantly greater than that during walking. Thigh extension and anterior pelvic tilt were significantly greater during running than walking. Also, a significant positive correlation was found between hip extension and anterior pelvic tilt during both walking and running, indicating that anterior pelvic tilt was greater in subjects that displayed reduced utilized peak hip extension. Thus, compensations for the increased stride length during running seem to occur at the pelvis, and presumably in the lumbar spine, rather than at the hip. Considering the association between anterior pelvic tilt and lumbar lordosis, the present findings may have clinical relevance regarding the prevention and treatment of hamstring injuries and of injuries to the lumbar spine. PMID- 19124246 TI - Cortical hyperexcitability and epileptogenesis: understanding the mechanisms of epilepsy - part 1. AB - Epilepsy encompasses a diverse group of seizure disorders caused by a variety of structural, cellular and molecular alterations of the brain primarily affecting the cerebral cortex, leading to recurrent unprovoked epileptic seizures. In this two-part review we examine the mechanisms underlying normal neuronal function and those predisposing to recurrent epileptic seizures starting at the most basic cellular derangements (Part 1) and working up to the highly complex epileptic networks (Part 2). We attempt to show that multiple factors can modify the epileptic process and that different mechanisms underlie different types of epilepsy, and in most situations there is an interplay between multiple genetic and environmental factors. PMID- 19124247 TI - A novel and potent VLA-4 antagonist based on trans-4-substituted cyclohexanecarboxylic acid. AB - During the course of our study, it was revealed that the poor pharmacokinetic properties of a series of benzoic acid derivatives such as 1 should be attributed to the diphenylurea moiety. Thus, we replaced the diphenylurea moiety in 1 with a 2-(2-methylphenylamino)benzoxazole moiety which mimics the diphenylurea structure. However, this modification resulted in a significant decrease (3, IC(50)=19 nM) in VLA-4 inhibitory activity compared to 1 (IC(50)=1.6 nM). To address this discrepancy, we worked on optimization of the carboxylic acid moiety in compound 3. As a result, our efforts have led to the discovery of trans-4 substituted cyclohexanecarboxylic acid derivative 11b (IC(50)=2.8 nM) as a novel and potent VLA-4 antagonist. In addition, compound 11b exhibited favorable pharmacokinetic properties (CL=3.3 ml/min/kg, F=51%) in rats. PMID- 19124248 TI - Death receptor 5 targeting activity-guided isolation of isoflavones from Millettia brandisiana and Ardisia colorata and evaluation of ability to induce TRAIL-mediated apoptosis. AB - Death receptor 5 (DR5) is an apoptosis-inducing membrane receptor for TNF-related apoptosis-inducing ligand (TRAIL). On screening for compounds that enhance DR5 expression using a luciferase assay with DLD-1/SacI, we previously identified 4' demethyltoxicarol isoflavone (1) isolated from the leaves of Millettia brandisiana. In this study, we revealed that 1 sensitized TRAIL-resistant human gastric adenocarcinoma (AGS) cells to TRAIL-induced apoptosis by up-regulating the expression of DR5. 1 induced DR5 expression at both the mRNA and protein level. A human recombinant DR5/Fc chimera remarkably inhibited 1-induced apoptosis. These results suggest that the enhancement of DR5 expression by 1 was critical to the cell death. Furthermore, a MeOH extract of the bark of Ardisia colorata markedly enhanced DR5 activity in this screening system. Bioassay-guided fractionation of A. colorata led to the isolation and identification of a new isoflavone, coloratanin A (3), together with ten known compounds. The chemical structure of the new compound was elucidated on the basis of a spectroscopic analysis. PMID- 19124249 TI - Cooperative alkylation of double-strand human telomere repeat sequences by PI polyamides with 11-base-pair recognition based on a heterotrimeric design. AB - We designed and synthesized alkylating conjugates 5-7 and their partner N methylpyrrole-N-methylimidazole (PI) polyamides 8, 9. The DNA alkylating activities of conjugates 5-7 were evaluated by high-resolution denaturing polyacrylamide gel electrophoresis with a 219 base pair (bp) DNA fragment containing the human telomere repeat sequence. Conjugate 5 efficiently alkylated the sequence, 5'-GGTTAGGGTTA-3', in the presence of partner PI polyamide 8 or distamycin A (Dist). In contrast, the heterodimer system of 5 with 9 showed very weak alkylating activity. Accordingly, this heterotrimeric system of 5 with two short partners is an expedient way to attain improved precision and extension of the recognition of DNA sequences. PMID- 19124250 TI - Novel DNA-directed alkylating agents: design, synthesis and potent antitumor effect of phenyl N-mustard-9-anilinoacridine conjugates via a carbamate or carbonate linker. AB - A series of phenyl N-mustard-9-anilinoacridine conjugates via a carbamate or carbonate linker was synthesized for antitumor evaluation. The carbamate or carbonate linker is able to lower the reactivity of the phenyl N-mustard pharmacophore and thus, these conjugates are rather chemically stable. The in vitro studies revealed that these derivatives possessed significant cytotoxicity with IC(50) in sub-micromolar range in inhibiting human lymphoblastic leukemia (CCRF-CEM), breast carcinoma (MX-1), colon carcinoma (HCT-116) and human non small cell lung cancer (H1299) cell growth in vitro. Compounds 10a, 10b, 10e, 10i, and 15a were selected for evaluating their antitumor activity in nude mice bearing MX-1 and HCT-116 xenografts. Remarkably, total tumor remission was achieved by these agents with only one cycle of treatment. Interestingly, no tumor relapse was found in mice treated with 10a over 129 days. This agent is capable of inducing DNA interstrand cross-linking in human non-small lung cancer H1299 cells in a dose dependent manner by modified comet assay and has a long half-life in rat plasma. PMID- 19124251 TI - Towards Gram-negative antivirulence drugs: new inhibitors of HldE kinase. AB - Gram-negative bacteria lacking heptoses in their lipopolysaccharide (LPS) display attenuated virulence and increased sensitivity to human serum and to some antibiotics. Thus inhibition of bacterial heptose synthesis represents an attractive target for the development of new antibacterial agents. HldE is a bifunctional enzyme involved in the synthesis of bacterial heptoses. Development of a biochemical assay suitable for high-throughput screening allowed the discovery of inhibitors 1 and 2 of HldE kinase. Study of the structure-activity relationship of this series of inhibitors led to highly potent compounds. PMID- 19124252 TI - Solid-phase parallel synthesis and SAR of 4-amidofuran-3-one inhibitors of cathepsin S: effect of sulfonamides P3 substituents on potency and selectivity. AB - Highly potent and selective 4-amidofuran-3-one inhibitors of cathepsin S are described. The synthesis and structure-activity relationship of a series of inhibitors with a sulfonamide moiety in the P3 position is presented. Several members of the series show sub-nanomolar inhibition of the target enzyme as well as an excellent selectivity profile and good cellular potency. Molecular modeling of the most interesting inhibitors describes interactions in the extended S3 pocket and explains the observed selectivity towards cathepsin K. PMID- 19124253 TI - Carbonic anhydrase inhibitors: inhibition of the beta-class enzyme from the yeast Saccharomyces cerevisiae with sulfonamides and sulfamates. AB - The protein encoded by the Nce103 gene of Saccharomyces cerevisiae, a beta carbonic anhydrase (CA, EC 4.2.1.1) designated as scCA, has been cloned, purified, characterized kinetically and investigated for its inhibition with a series of sulfonamides and one sulfamate. The enzyme showed high CO(2) hydrase activity, with a k(cat) of 9.4x10(5)s(-1), and k(cat)/K(M) of 9.8x10(7)M(-1)s( 1). Simple benzenesulfonamides substituted in 2-, 4- and 3,4-positions of the benzene ring with amino, alkyl, halogeno and hydroxyalkyl moieties were weak scCA inhibitors with K(I)s in the range of 0.976-18.45 microM. Better inhibition (K(I)s in the range of 154-654 nM) was observed for benzenesulfonamides incorporating aminoalkyl/carboxyalkyl moieties or halogenosulfanilamides; benzene 1,3-disulfonamides; simple heterocyclic sulfonamides and sulfanilyl-sulfonamides. The clinically used sulfonamides/sulfamate (acetazolamide, ethoxzolamide, methazolamide, dorzolamide, topiramate, celecoxib, etc.) generally showed effective scCA inhibitory activity, with K(I)s in the range of 82.6-133 nM. The best inhibitor (K(I) of 15.1 nM) was 4-(2-amino-pyrimidin-4-yl) benzenesulfonamide. These inhibitors may be useful to better understand the physiological role of beta-CAs in yeast and some pathogenic fungi which encode orthologues of the yeast enzyme and eventually for designing novel antifungal therapies. PMID- 19124254 TI - Fabrication and characterization of TiN-Ag nano-dice. AB - TiN-Ag nanocomposite was synthesized by dc arc-plasma method. Microstructures of TiN-Ag nanocomposite were carefully characterized by powder X-ray diffraction method and transmission electron microscopy, and nano-morphologies by three dimensional electron tomography. It was found that the surface of nanocrystalline TiN matrix was densely covered by finely dispersed Ag nanoparticles, and it was found that they were physically attached but not chemically bonded from their orientation relationships. PMID- 19124255 TI - Low-temperature effect on enzyme activities involved in sucrose-starch partitioning in salt-stressed and salt-acclimated cotyledons of quinoa (Chenopodium quinoa Willd.) seedlings. AB - The effect of low temperature on growth, sucrose-starch partitioning and related enzymes in salt-stressed and salt-acclimated cotyledons of quinoa (Chenopodium quinoa Willd.) was studied. The growth of cotyledons and growing axes in seedlings grown at 25/20 degrees C (light/dark) and shifted to 5/5 degrees C was lower than in those only growing at 25/20 degrees C (unstressed). However, there were no significant differences between low-temperature control and salt-treated seedlings. The higher activities of sucrose phosphate synthase (SPS, EC 2.4.1.14) and soluble acid invertase (acid INV, EC 3.2.1.25) were observed in salt-stressed cotyledons; however, the highest acid INV activity was observed in unstressed cotyledons. ADP-glucose pyrophosphorylase (ADP-GPPase, EC 2.7.7.27) was higher in unstressed cotyledons than in stressed ones. However, between 0 and 4days the highest value was observed in salt-stressed cotyledons. The lowest value of ADP GPPase was observed in salt-acclimated cotyledons. Low temperature also affected sucrose synthase (SuSy, EC 2.4.1.13) activity in salt-treated cotyledons. Sucrose and glucose were higher in salt-stressed cotyledons, but fructose was essentially higher in low-temperature control. Starch was higher in low-temperature control; however, the highest content was observed at 0day in salt-acclimated cotyledons. Results demonstrated that low temperature induces different responses on sucrose starch partitioning in salt-stressed and salt-acclimated cotyledons. Data also suggest that in salt-treated cotyledons source-sink relations (SSR) are changed in order to supply soluble sugars and proline for the osmotic adjustment. Relationships between starch formation and SuSy activity are also discussed. PMID- 19124256 TI - Electron emission from photo-excited testosterone in water-ethanol solution. AB - Testosterone (TES; 4-androstene-17beta-ol-3-on) is found for the first time to eject electrons from its singlet excited state in water-ethanol solvent mixture. This ability was very recently also observed for 17beta-estradiol (17betaE2) and progesterone (PRG)/1/. With increasing TES-concentration, the yield of solvated electrons (e(s)(-)) is decreasing, because of "associate" formation. At higher absorbed UV-doses (lambda=254 nm) the e(s)(-) yield is passing a sharp maximum by formation of TES-ethanol adducts, which are able likewise to emit electrons when excited. At prolonged irradiation the resulting photolytic products of TES ethanol adducts are also able to emit electrons. The capability of the hormones: 17betaE2, PRG and TES to eject electrons and the resulting metabolites, some of which can induce cancer, is discussed. PMID- 19124257 TI - Enantiomeric behaviour of albendazole and fenbendazole sulfoxides in domestic animals: pharmacological implications. AB - Albendazole and fenbendazole are methylcarbamate benzimidazole anthelmintics extensively used to control gastrointestinal parasites in domestic animals. These parent compounds are metabolised to albendazole sulfoxide and fenbendazole sulfoxide (oxfendazole), respectively. Both sulfoxide derivatives are anthelmintically active and are manufactured for use in animals. They metabolites have an asymmetric centre on their chemical structures and two enantiomeric forms of each sulfoxide have been identified in plasma, tissues of parasite location and within target helminths. Both the flavin-monooxygenase and cytochrome P450 systems are involved in the enantioselective biotransformation of these anthelmintic compounds in ruminant species. A relevant progress on the understanding of the relationship among enantioselective metabolism and systemic availability of each enantiomeric form has been achieved. This article reviews the current knowledge on the pharmacological implications of the enantiomeric behaviour of albendazole sulfoxide and oxfendazole in domestic animals. PMID- 19124258 TI - Effect of conformational degrees of freedom on the charge transfer in model tripeptide. AB - An extensive conformational dependence of the intramolecular charge transfer (both hole and electron) between intermediate residues of the model tripeptide in gas phase has been studied. The charge transfer integral, spatial overlap integral and site-energy for both hole and electron transfer between the intermediate residues in the tripeptides were calculated using the fragment orbital method. The site-energies and the charge transfer integrals have been calculated for different conformation of the glycine tripeptide by varying the dihedral angles (phi and psi) along the alpha-carbon atom of amino acid subgroups. Electronic structure calculations show that the charge transfer integral between intermediate residues is strongly depending on the nature of the conformation of the peptide. The calculations indicate that the charge transfer is maximum at the particular conformation of the intermediate amino acid residues. PMID- 19124261 TI - Endothelial microparticles in patients with insulin resistance. PMID- 19124260 TI - A paradoxical reaction during antituberculosis therapy for congenital tuberculosis. AB - A paradoxical tuberculosis (TB) reaction is defined as the clinical or radiological worsening of pre-existing TB lesions or the development of new lesions during treatment. We treated a case of such a paradoxical reaction in a 21-day-old female infant who was diagnosed with congenital TB and was being treated with antituberculous drugs. The paradoxical reaction improved after additional treatment with corticosteroids. Therefore, corticosteroids might be useful to control paradoxical reactions in patients with congenital tuberculosis. PMID- 19124259 TI - Postgraduate institute management protocol for invasive Aspergillus flavus sinusitis: is it effective? AB - OBJECTIVE: To analyze the results of a postgraduate institute management protocol for invasive Aspergillus flavus sinusitis. METHODS: This was a prospective study wherein all patients with a final histopathological diagnosis of invasive sino orbital aspergillosis were included for the period between January 1998 and December 2005. All the patients were negative for HIV, diabetes, and chronic granulomatous disease. All patients were evaluated clinically, biochemically, and radiologically for the condition, and were classified into four groups on the basis of clinical and radiological findings. All patients were subjected to endonasal endoscopic biopsy and were subjected as per the management protocol. These patients were then followed up clinically, endoscopically, and radiologically for a period ranging from 9 months to 7 years. RESULTS: A total of 74 patients were enrolled in the study, 52 males and 22 females, ranging in age from 26 to 53 years. The most common sign was proptosis. The patients in each group were subjected to the treatment protocol decided for the group. Overall, residual disease was seen in 14 patients (18.9%) and recurrence was seen in six patients (8.1%). Twenty-four (32.4%) patients died of the disease in our series. Residual disease and recurrent disease were seen more often in the advanced cases. CONCLUSION: Invasive sino-orbital aspergillosis is associated with significant mortality. By following our management protocol we were able to reduce the mortality to 32.4% in our series. PMID- 19124262 TI - [An enlarged dismantable anatomical model of the ear by Auzoux: observation and photographical tridimensional reconstruction]. AB - The aim of the study was to determine the accuracy of a 19th century anatomical model of an ear by Auzoux (a French manufacturer). It measured 63 cm in length; 43 cm in width; 37 cm in height. The disassembled parts were studied on the morphological aspect in its whole and after removing of its components: external ear (43 cm x 23 cm x 15 cm), tympanic drum and middle ear ossicles; cochleo vestibular apparatus. The main dimensions of each part were recorded. The arterial vascularisation of the three components of the ear and its sensitive and sensorial nervous system were established on the model which represented the external carotid artery with its terminal bifurcation, several collateral branches specially the posterior auricular artery and the middle meningeal artery; the internal carotid artery at the two extremities of the carotid canal. The cochleo-vestibular and facial nerve as well as other elements of small calibre were exactly figured and labelled by a number. The middle ear components were disassembled. The ossicles of the middle ear and a dried peritoneal tympanic membrane were dismountable as a whole. Only a few minor modification from the admitted data of anatomy as well as the cutaneous muscles of the external ear were observed on this sample, the anatomical aspects of which were very precisely represented. The material was typical of the Auzoux manner: a special papier mache whose conception and realization remains unequalled. This kind of very rare model can be dated at the earliest 1835 (Lemire, 1990). PMID- 19124264 TI - [Pathological fracture of the carpal scaphoid (intra-osseous synovial cyst). Clinical and therapeutic aspect. Case report]. AB - The authors report a case of fracture of the carpal scaphoid through an intraosseous synovial cyst. Synovial cysts are not uncommon in the carpal region, and are usually discovered as an incidental radiological finding: they rarely present with wrist pain and only exceptionally as a fracture. In our knowledge, only one other case of intraosseous synovial cyst of the carpal scaphoid presenting as a fracture has been published. Surgical treatment of these benign tumors, in cases of fracture or painful symptoms, consists of curettage of the lesion followed by impaction grafting of cancellous bone combined with stabilization in the case of a fracture. The use of a vascularized bone graft pedicled on the volar carpal artery enables simultaneous treatment of both the cyst and the fracture. The prognosis is good and relapse is exceptional. PMID- 19124263 TI - Familial chilblain and late contractural arachnodactyly: a novel association? AB - We report an Italian family suffering from chilblain. Seven members over three generations affected, two of them presenting association with late contractural arachnodactyly. Inflammatory cutaneous lesions following exposure to cold or drop in temperature are called chilblain. Arachnodactyly is an hallmark of hereditary connective tissue disorders such as Marfan's syndrome and Beals' syndrome. The latter, showing joint congenital contractures, is called indeed congenital contractural arachnodactyly. We speculate that an underlying alteration in connective tissue could lead to the association between chilblain lesions and contractural arachnodactyly. PMID- 19124265 TI - [Dorsally displaced intra-articular distal radius fractures treated by locked palmar plate. Technique and results]. AB - OBJECTIVES: The purpose of this study was to evaluate the results and complications of locked palmar plating in patients with articular fracture of the distal radius. METHODS: Twenty-two patients were reviewed retrospectively. The average age was 68 years. All fractures were classified as Type C according to the AO classification. Clinical and functional examination including range of motion, grip strength, pain and return to previous activities were assessed. Loss of radial height, radial inclination, palmar tilt and ulnar variance were evaluated with preoperative and postoperative radiographs. RESULTS: At an average follow-up of 10 months, the range of motion was 124 degrees in flexion and extension and 178 degrees in pronation and supination. Grip strength was 80% of the opposite side. Fifteen patients were free of pain. Three patients suffered tenosynovitis of the extensor tendons. Radiographic measurements averaged 11 mm radial height, 21 degrees radial inclination, 4 degrees palmar tilt and 1 mm ulnar variance. CONCLUSIONS: The result of this study showed that locked palmar plating of articular distal radius fractures is effective even in cases of metaphyseal comminution with a low rate of complications. PMID- 19124266 TI - Threshold curves obtained under various gaseous conditions for free radical generation by burst ultrasound - Effects of dissolved gas, microbubbles and gas transport from the air. AB - To understand the underlying concepts required for the determination of thresholds for free radical generation, effects of gas dissolution in and microbubble addition to sonicated solutions were investigated. Four solutions with different gaseous conditions, air-saturated and degassed solutions with and without microbubbles of 20 microm in diameter with shells, were studied in the presence of an air-liquid interface. These test solutions were exposed to 1 MHz ultrasound of 0.06 MPa(p-p) at various pulse durations (PDs) from 0.1 to 5 ms and pulse repetition frequencies from 0.1 to 2 kHz. Generation of free radicals was evaluated using the electron spin resonance (ESR) spin trapping method and starch iodine method. Thresholds of duty ratio (DR) corresponding to temporal average intensity of ultrasound for free radical generation were significantly greater in degassed solutions than in air-saturated solutions. Microbubbles had no significant effects in air-saturated solutions but caused a slight decrease in the threshold in degassed solutions. In all of these results, the DR of a threshold curve against pulse repetition period (PRP) was not constant but linearly decreased with it, suggesting that a balance between bubble growth and shrinkage during the ON and OFF times of burst ultrasound is the primary parameter for the interpretation of thresholds. The effect of an air-liquid interface of the solution was also examined, and it was revealed that gas transport from the air is a predominant factor determining the amount of free radicals. PMID- 19124267 TI - Modelling of experimentally created partial-thickness human skin burns and subsequent therapeutic cooling: a new measure for cooling effectiveness. AB - Rapid post-injury cooling of a skin burn has been shown to have both symptomatic and therapeutic benefits. However, the latter cannot be explained by temperature reduction alone, and must thus be secondary to an altered biological response. In this study, we construct a computational model to calculate the heat transfer and damage accumulation in human skin during and after a burn. This enables us to assess the effectiveness of various cooling protocols (involving both free and forced convection to air and water respectively) in terms of their reduction in Arrhenius tissue damage. In this process, we propose an extension of the Arrhenius damage model in the form of a new measure xi, which estimates the relevance of post-burn accrued damage. It was found that the reduction in Arrhenius damage integrals near the skin surface was too small to be physiologically relevant. Hence our results confirm that while the reduction in tissue temperatures is indeed quicker, the therapeutic benefit of cooling cannot be explained by thermal arguments (i.e. based on Arrhenius damage models) alone. We plan to validate this hypothesis by conducting future microarray analyses of differential gene expression in cooled and non-cooled burn lesions. Our computational model will support such experiments by calculating the necessary conditions to produce a burn of specified severity for a given experimental setup. PMID- 19124268 TI - Synthesis, DNA binding and cleavage studies of Ni(II) complexes with fused aromatic N-containing ligands. AB - The three Ni(II) complexes of fused aromatic N-containing ligands such as [Ni(bnp)(3)](PF(6))(2) (1), [Ni(phen)(2)(bnp)](PF(6))(2) (2) and [Ni(bpy)(2)(bnp)](PF(6))(2) (3) (where bnp=dibenzo(b)1,8-naphthpyridine, phen=1,10-phenanthroline and bpy=bipyridine) were synthesized and structurally characterized. Elemental analysis, magnetic and spectroscopic data suggested octahedral geometry for all the complexes. Binding of these complexes with (ds)DNA were analyzed by absorption spectra, viscosity and thermal denaturation studies. Detailed analysis revealed that the metal complexes intercalates into the DNA base stack as intercalator. The oxidative cleavage activities of the complexes were studied with supercoiled (SC)pUC19 DNA by using gel electrophoresis, and the results show that complexes have potent nuclease activity. PMID- 19124269 TI - Analysis of vibrational spectra of 1-chloro-2,4-dinitrobenzene based on density functional theory calculations. AB - This work deals with the vibrational spectroscopy of 1-chloro-2,4-dinitrobenzene (CDNB) by means of quantum chemical calculations. The FTIR and FT-Raman spectra were measured in the condensed state. The fundamental vibrational frequencies and intensity of vibrational bands were evaluated using density functional theory (DFT) with the standard B3LYP/6-311+G** basis set combinations, and was scaled using various scale factors which yields a good agreement between observed and calculated frequencies. The vibrational spectra were interpreted with the aid of normal coordinate analysis based on scaled quantum mechanical force field. The results of the calculations were applied to simulated infrared and Raman spectra of the title compound, which showed excellent agreement with the observed spectra. PMID- 19124270 TI - Vibrational spectroscopic study of terbutaline hemisulphate. AB - The Raman spectrum of terbutaline hemisulphate is reported for the first time, and molecular assignments are proposed on the basis of ab initio BLYP DFT calculations with a 6-31G* basis set and vibrational frequencies predicted within the quasi-harmonic approximation; these predictions compare favourably with the observed vibrational spectra. Comparison with previously published infrared data explains several spectral features. The results from this study provide data that can be used for the preparative process monitoring of terbutaline hemisulphate, an important beta(2) agonist drug in various dosage forms and its interaction with excipients and other components. PMID- 19124271 TI - Spectroscopic analysis and DFT calculations of a food additive carmoisine. AB - FT-IR and Raman techniques were employed for the vibrational characterization of the food additive Carmoisine (E122). The equilibrium geometry, various bonding features, and harmonic vibrational wavenumbers have been investigated with the help of density functional theory (DFT) calculations. A good correlation was found between the computed and experimental wavenumbers. Azo stretching wavenumbers have been lowered due to conjugation and pi-electron delocalization. Predicted electronic absorption spectra from TD-DFT calculation have been analysed comparing with the UV-vis spectrum. The first hyperpolarizability of the molecule is calculated. Intramolecular charge transfer (ICT) responsible for the optical nonlinearity of the dye molecule has been discussed theoretically and experimentally. Stability of the molecule arising from hyperconjugative interactions, charge delocalization and C-H ...O, improper, blue shifted hydrogen bonds have been analysed using natural bond orbital (NBO) analysis. PMID- 19124272 TI - Self-reported skill level and injury severity in skiers and snowboarders. AB - There is evidence to suggest that the rate of injury is lower for expert skiers and snowboarders than for beginners. A better understanding of the relation between injury severity and skill level is also needed for planning injury prevention strategies. Our objective was to examine the severity and location of injuries sustained by self-reported expert and beginner skiers and snowboarders. A case-control study design was used. Injured skiers and snowboarders had to report their skill level on a 5 point scale (1: "beginner"; 5: "expert"). Two sets of severely injured cases were defined based on the type of injury and ambulance evacuation. Controls were those who did not sustain severe injuries. Logistic regression analyses were performed to relate injury severity to skill level. Subjects were 22 078 injured skiers and snowboarders who reported to the ski patrol with an injury sustained on the slopes of an alpine ski centre of the Canadian province of Quebec during the seasons 2001-2002 to 2004-2005. Compared with beginners, experts had an increased risk of suffering from a severe injury (adjusted odds ratio [AOR]: 1.88; 95% CI: 1.58-2.23). Expert snowboarders were also more likely to suffer from a severe injury or be evacuated by ambulance (AOR: 1.18; 95% CI: 1.02-1.38). Results suggest that the type of activities or manoeuvres performed by expert skiers and snowboarders may increase the risk of sustaining a severe injury compared with beginner participants. PMID- 19124273 TI - The challenge of understanding and assessing physical activity in preschool-age children: Thinking beyond the framework of intensity, duration and frequency of activity. AB - The physical activity behavior of young preschool-aged children is different from that seen in older children and adults, due in part to biological, psychosocial and cognitive immaturity. This paper: (a) provides an overview the relevant development of young children in relation to physical activity; and (b) outlines the relationship between child development, play, and physical activity in young children. Understanding, assessing and promoting physical activity in young children should include identification of contextual factors such as developmental stage and aspects of play. PMID- 19124274 TI - The kinin B(1) receptor and inflammation: new therapeutic target for cardiovascular disease. AB - The kinin B(1) receptor plays an important role in mediating the inflammatory effects of the kallikrein-kinin pathway. The recent development of orally available non-peptidic antagonists and genetically modified mice deficient in B(1) receptor expression have demonstrated that the receptor plays a pivotal role in the cellular, particularly neutrophil, recruitment associated with an acute inflammatory response. These tools have also enabled elucidation of the pathways involved in mediating this effect and have highlighted a major role for chemokines, particularly CXCL5 and CCL2. Neutrophil recruitment is involved in the pathogenesis of renal disease and has very recently been implicated in the early stages of atherosclerosis. In this review we discuss the most recent evidence linking the B(1) receptor with the pathogenesis of these two inflammatory cardiovascular diseases and highlight the therapeutic potential of the kinin B(1) receptor in these disease states. PMID- 19124275 TI - Podcasts and videostreaming: Useful tools to facilitate learning of pathophysiology in undergraduate nurse education? AB - Nurses require an understanding of pathophysiological processes to support safe practice in the clinical area. A review of the literature indicates that nursing students have traditionally found this to be a challenging element of the course. The increasing availability of newer technology in higher education such as videostreaming and podcasting provides the opportunity to utilise a variety of approaches to cater for a wider range of learning styles. In view of this, lecturers developed multimedia resources to incorporate into a pathophysiology module. An evaluation of students' views on the use of ICT to facilitate understanding of pathophysiology was undertaken. The majority of students appeared to welcome the incorporation of alternative teaching approaches into the module. Interestingly despite the opportunities which technology offers to enhance learning many of the students also highlighted the importance of maintaining significant face-to-face contact with lecturers. This supports the findings of the literature which indicates that students find hybrid learning strategies that combine traditional teaching with more innovative methods particularly beneficial. PMID- 19124276 TI - Size-dependent palindrome-induced intrachromosomal recombination in yeast. AB - Palindromic and quasi-palindromic sequences are important DNA motifs found in various cis-acting genetic elements, but are also known to provoke different types of genetic alterations. The instability of such motifs is clearly size related and depends on their potential to adopt secondary structures known as hairpins and cruciforms. Here we studied the influence of palindrome size on recombination between two directly repeated copies of the yeast CYC1 gene leading to the loss of the intervening sequence ("pop-out" recombination). We show that palindromes inserted either within one copy or between the two copies of the CYC1 gene become recombinogenic only when they attain a certain critical size and we estimate this critical size to be about 70 bp. With the longest palindrome used in this study (150 bp) we observed a more than 20-fold increase in the pop-out recombination. In the sae2/com1 mutant the palindrome-stimulated recombination was completely abolished. Suppression of palindrome recombinogenicity may be crucial for the maintenance of genetic stability in organisms containing a significant number of large palindromes in their genomes, like humans. PMID- 19124277 TI - Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization. AB - Analytical derivatization (AD) increases the sensitivity of analysis by one to three orders of magnitude, stabilizes labile analytes and converts them into readily extractable products. Using a variant of this technique, we applied solid phase analytical derivatization (SPAD) to fully automate extraction, derivatization and liquid chromatography. The resulting device (AutoSPAD) determined malonyldialdehyde (MDA) from biological fluids. This biomarker of oxidative stress is highly water-soluble (500 g/L at pH 7), chemically labile and lacks any functionality that enables detection at high sensitivity. AutoSPAD utilizes column-switching technology to load DANSYL hydrazine onto the solid phase, pass the biological sample over the resulting reactor bed for derivatization on the surface to form a hydrophobic derivative suitable for increasing sensitivity of any other LC technique including LC-MS/MS. The hydrophobic solid phase retains the derivative during washing steps, following which AutoSPAD transfers the derivatized extract to the analytical column for separation and detection by fluorescence. In plasma, however, MDA exists both in free form and covalently bound to protein. Measuring MDA from plasma, therefore, required identification of appropriate protein precipitation and hydrolysis conditions. Under these conditions, the DANSYL derivative formed at only one aldehydic position but did not cyclize as reported for other reactions between hydrazine reagents and MDA. The calibration curve using approximately 7 microL of plasma was linear (r(2)=0.999) in the physiological range (0.1-3 microg/mL) and the relative standard deviation of replicate determinations at 1 microg/mL was less than 5%. PMID- 19124278 TI - Absolute ProGRP quantification in a clinical relevant concentration range using LC-MS/MS and a comprehensive internal standard. AB - The objective of this study was to develop a method using liquid chromatography with tandem mass spectrometric detection for the absolute quantification of the small cell lung cancer biomarker ProGRP in human serum, using its tryptic signature peptide NLLGLIEAK. The samples were precipitated for most of its proteins using acetonitrile prior to tryptic digestion. Further sample clean-up and enrichment was achieved by the use of an on-line restricted access media column, followed by separation on a BioBasic C8 column. Detection and quantification was carried out by operating a triple quadrupole MS in the selected reaction monitoring mode. This setup allowed analysis of realistic samples and detections limits in human serum of 150 pg ProGRP on column. Using an internal standard derived from the parent ProGRP after acetylation of the lysine side chain allowed better quantification through variation correction in all sample pretreatment steps, trypsination included. PMID- 19124279 TI - LC-MS/MS and centrifugal ultrafiltration method for the determination of novobiocin in chicken, fish tissues, milk and human serum. AB - We present a rapid and simple method for detecting novobiocin in biologic samples using a methanol-based extraction of the tissue matrix and liquid chromatography with electrospray tandem mass spectrometry (LC-ESI-MS/MS) on positive mode. The sample, prepared using centrifugal ultrafiltration with 5.0% SDS, was directly injected into the LC-MS/MS. Chromatographic separation was performed on a TSK-GEL ODS 100 V column using 0.5% formic acid in water/methanol. The method was validated according to the Japanese Maximum Residue Limits recommendations. Detection was linear over a range of 5-100 ppb matrix solution (r>0.998). Novobiocin recovery values from chicken (0.05 ppm) and fish tissues (0.05 ppm), milk (0.08 ppm), and human serum (0.05 and 0.01 ppm) samples ranged from 71+/-1 to 95+/-2%. PMID- 19124280 TI - Development of an LC-ESI-MS/MS method for the determination of histamine: application to the quantitative measurement of histamine degranulation by KU812 cells. AB - A rapid, simple, and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for the identification and quantification of histamine without a previous derivatization step or the addition of general ion-pairing reagents to the mobile phase. This method was used to measure histamine release following degranulation of KU812 human basophilic cells, using pyrazol as an internal standard. Analyses were performed on an LC system employing a Cosmosil 5C(18) PAQ column and an isocratic elution with methanol-0.005% trifluoroacetic acid (1:1) at a flow rate of 0.2 mL/min. A triple-quadrupole mass spectrometer, equipped with an electrospray ionization interface was employed, operating in the positive ion mode. The retention time of histamine and the internal standard were 4.0 and 5.0 min, respectively. The relative standard deviations (R.S.D.s) of the retention time and peak area were between 0.47% and 2.03%. Micropipette tip solid-phase extraction (SPE) using LooseTip C(18) allowed for not only rapid sample preparation, but also decreased suppression effects, improving peak shape. This method was used to evaluate the anti-allergic effects of compounds contained in Taxus yunnanensis extracts. Four constituents that were isolated from the wood extracts of T. yunnanensis and sodium cromoglicate, which is used as a first line anti-allergic drug, were tested in an in vitro histamine release inhibition assay. Of these compounds, taxiresinol and isotaxiresinol were more inhibitory than sodium cromoglicate. PMID- 19124281 TI - Development and validation of a LC/MS/MS method for quantification of nobiliside A in rat plasma. AB - A LC/MS/MS method was developed and validated for determination of nobiliside A in rat plasma. Analyses were separated by an Agella Venusil ASB-C18 column and isocratic elution with methanol:water (80:20, v/v) as a mobile phase. A tandem mass spectrometer was used as a detector for quantitative analysis. Calibration curves (R(2)>0.999) in spiked plasma were linear over the concentration range of 50-5000 ng/mL. The overall accuracy of this method was 93-104% for nobiliside A. Within-day and between-day variability was both less than 9% in plasma. The data indicate that the LC/MS/MS method is an effective method for the pharmacokinetics study of nobiliside A in rat plasma. PMID- 19124282 TI - Determination of sulfadimethoxine and 4N-acetylsulfadimethoxine in bovine plasma, urine, oral fluid, and kidney and liver biopsy samples obtained surgically from standing animals by LC/MS/MS. AB - A quantitative method was developed and validated to measure the concentration of sulfadimethoxine (SDM) and its major metabolite, (4)N-acetylsulfadimethoxine (AcSDM), in bovine tissues and body fluids. Liquid chromatography/tandem mass spectrometry (LC/MS/MS) gave quantitative results for these two analytes in extracts from bovine plasma, urine, oral fluid, kidney, and liver, using SDM-d(4) as internal standard (I.S.). The lower limit of quantitation (LLOQ) for both analytes in these matrices was validated at 2, 100, and 5 ng/mL in plasma, urine, and oral fluid respectively, and 10 ng/g in both kidney (cortex) and liver. The overall accuracy (average of 4 levels) is, for plasma, 104% (SDM) and 95% (AcSDM), with standard deviation of 9% (SDM) and 15% (AcSDM); for urine, 100% (SDM) and 106% (AcSDM), with standard deviation of 5% (SDM) and 6% (AcSDM); for oral fluid, 103% (SDM) and 103% (AcSDM), with standard deviation of 4% (SDM) and 4% (AcSDM); for kidney, 101% (SDM) and 111% (AcSDM), with standard deviation of 7% (SDM) and 6% (AcSDM); and for liver, 99% (SDM) and 115% (AcSDM), with standard deviation of 11% (SDM) and 9% (AcSDM). C18 SPE cartridges were used to clean-up these matrices, except for urine which was diluted directly with buffer before analysis by LC/MS/MS. PMID- 19124283 TI - Enzymatic diagnosis of Sjogren-Larsson syndrome using electrospray ionization mass spectrometry. AB - BACKGROUND: Sjogren-Larsson syndrome is a metabolic disorder characterized by accumulation of long-chain fatty alcohols in plasma of patients due to mutations in the ALDH3A2 gene, that codes for a microsomal fatty aldehyde dehydrogenase (FALDH). Recent studies have demonstrated that FALDH is involved in the last step of the conversion of 22-hydroxy-C22:0 into the dicarboxylic acid of C22:0 (C22:0 DCA). METHODS: FALDH activity was determined by incubating fibroblast homogenates with omega-hydroxy-C22:0 in the presence of NAD(+). Electrospray ionization mass spectrometry (ESI-MS) was used to quantify the amounts of C22:0-DCA produced. RESULTS: All SLS patients were deficient in C22:0-DCA productions with activities ranging from 3.2-26.3% of mean control. CONCLUSIONS: The new assay described in this paper has substantial advantages over previous assays, and allows for the easy, reliable and rapid diagnosis of SLS. PMID- 19124284 TI - An liquid chromatography-tandem mass spectrometry assay for determination of trace amount of new antifungal drug iodiconazole in human plasma. AB - A simple, rapid and sensitive LC-MS/MS method for determination of trace amount of new antifungal drug iodiconazole in human plasma was developed (1-(1H-1,2,4 triazole)-2-(2,4-diflurophenyl)-3-[N-methyl-N-(3-chlor-benzyl)amino]-2-propanol), was used as internal standard (IS). The analytes were extracted by liquid-liquid extraction with n-hexane after internal standard spiked. The separation was performed by a ZORBAX SB-C(18) column (3.5 microm, 2.1 mm x 100 mm) with an isocratic mobile phase consisting of methanol-water-formic acid (50:50:0.05, v/v/v) at a flow rate of 0.3 mL/min. The lower limit of quantification (LLOQ) was 0.10 ng/mL. This method was successfully used to determine the concentration of iodiconazole in human plasma following dermal administration. PMID- 19124285 TI - Validation of a sensitive gas chromatographic-mass spectrometric method for the simultaneous determination of beta-elemene and beta-elemenal in human plasma. AB - A sensitive gas chromatographic-mass spectrometric assay was described for determination of beta-elemene and beta-elemenal in human plasma, which has been successfully applied in clinical trial. After liquid-liquid extraction and gas chromatographic separation, the analytes were identified and quantitated. Calibration curves were linear in range from 31.25 to 8000 ng mL(-1) and the limit of quantification for both was 31.25 ng mL(-1). Intra- and inter-day precision at three concentrations were 2.3-8.3% with accuracy of -8.5 to 6.1% for elemene and 3.0-9.9% with accuracy of -2.3 to 5.9% for elemenal. The method was validated to be suitable for further pharmacokinetic study. PMID- 19124286 TI - Carrier mediated hollow fiber liquid phase microextraction combined with HPLC-UV for preconcentration and determination of some tetracycline antibiotics. AB - In the present study, a simple and efficient preconcentration method was developed using carrier mediated three phase liquid phase microextraction prior to HPLC-UV for simultaneous extraction and determination of trace amounts of highly hydrophilic tetracycline antibiotics including tetracycline (TCN), oxytetracycline (OTCN) and doxycycline (DCN) in bovine milk, human plasma and water samples. For extraction, 11.0 mL of the aqueous sample containing TCNs and 0.05 M Na(2)HPO(4) (9.10.995). Finally, applicability of the proposed method was successfully confirmed by extraction and determination of the drugs in water and plasma samples as well as in bovine milk samples with low and high fat contents. Comparing to the traditional methods, the proposed method exhibits high sensitivity and high preconcentration factors as well as good precision. The extraction setup is simple and due to active transport of analytes, high cleanup effect and good selectivity are obtained in extraction process. PMID- 19124287 TI - Bioassay guided discovery of apoptosis inducers from gamboge by high-speed counter-current chromatography and high-pressure liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry. AB - A screening system, composed of high-speed counter-current chromatography and high-pressure liquid chromatography/electrospray ionization quadrupole time-of flight mass spectrometry, was established to find bioactive lead compound. This system succeeded in discovering apoptosis inducers from gamboge, the resin of Garcinia hanburyi. High-speed counter-current chromatography was used to provide well-separated fractions for bioassay and the resulted active fractions were rapidly identified using high-pressure liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry. The solvent system of n hexane/ethyl acetate/methanol/water was optimized to the ratio of 7:3:7:3 (v/v/v/v) by a K value analysis. As a result, two active fractions were obtained. They showed apoptosis inducing effects as potent as that of taxol (500 nM) at the concentration of 1 microg/ml. Gambogenic acid (72.1%) and epimeric isogambogic acids (25.3%) were identified in one of the fractions. The other active fraction mainly contained two epimeric mixtures, gambogic acids (68.7%) and gambogoic acids (26.9%). Among them, gambogenic acid, without epimerization, has priority to be lead compound. PMID- 19124288 TI - Comment on 'The 'Gallipot' splash guard: a simple and effective safety measure for wound irrigation'. PMID- 19124289 TI - Congenital infiltrating lipomatosis. AB - The congenital infiltrating lipomatosis (LCI) represents a distinct, clinical and pathological aspect of fat soft-tissue neoplasia; very few cases of which have been described in literature. In 1983, Slavin and Cols described the main characteristics of LCI as:(1) non-encapsulated tumours containing mature adipocites, (2) fat infiltration in muscles and adjacent soft parts, (3) absence of malignant characteristics, (4) absence of lipoblasts, (5) presence of fibrous elements, increased number of vessels and nerves and (6) adjacent bone hypertrophy. In this article, we describe a congenital infiltrating lipomatosis of the face in a child and discuss the diagnoses and the treatment of this disease. PMID- 19124290 TI - Biomechanical stability analysis of rigid intraoral fixation for bilateral sagittal split osteotomy. AB - BACKGROUND: Biomechanical stability in patients in whom mandibular prognathism was corrected with different fixation methods during bilateral sagittal split osteotomy (BSSO) surgery remains controversial and needs to be clarified. METHODS: A three-dimensional (3D) finite element (FE) model of the mandible was developed to simulate the biomechanical responses of osteo-synthesis screws and the stability of different screw-placement arrangements in BSSO. Six types of fixation methods for the osseous segments were simulated with two or three screws in different placement arrangements to avoid injury to the inferior alveolar nerve. RESULTS: A triangular configuration of the screw position across the nerve presented less stress loading than the linear configuration, and hence provided better stability as the preferred fixation method for BSSO of the mandible. When the screws were aligned in a linear setting, the stress values were 4 times higher, implying a less stable fixation. Neither two nor three screws applied at the superior border appeared to be better at exploiting the increased thickness of the cortical bone encountered in this region. CONCLUSIONS: According to the 3D FE analysis, the configuration with three screws inserted in a triangular shape across the inferior alveolar nerve (Type 4) demonstrates the best rigidity among six screw-placement configurations. Three 2.3-mm diameter bi-cortical screws were considered a sufficient fixation tool after BSSO of the mandible. PMID- 19124291 TI - Successful re-construction of a large tissue defect using a free pre-fabricated flap with purely implanted arterialised venous loop. AB - Flap pre-fabrication represents an interesting technique in tissue-defect reconstruction. In this article we report the successful clinical application of a free pre-fabricated flap using an isolated arterialised venous loop as an implanted vascular pedicle for reconstruction of an extended soft-tissue defect on the leg, as well as clinical neo-vascularisation development following arterialised venous loop implantation. The procedure appears potentially useful in selected patients due to the easy harvest of an isolated vein loop as well as a favourable functional and aesthetic result at the donor site. PMID- 19124292 TI - Properties of cytochrome bd plastoquinol oxidase from the cyanobacterium Synechocystis sp. PCC 6803. AB - In the aerobic respiratory chain of the cyanobacterium Synechocystis sp. PCC 6803, cytochrome c oxidase serves as a major terminal oxidase while cyanide resistant cytochrome bd serves as an alternative oxidase and evades the over reduction of the plastoquinone pool under stress conditions. Here we expressed Synechocystis cytochrome bd in Escherichia coli and characterized enzymatic and spectroscopic properties. Cyanobacterial cytochrome bd showed the higher activity with ubiquinols than with decyl-plastoquinol and K(m) values for quinols were 2 fold smaller than those of E. coli cytochrome bd (CydAB). The dioxygen reduction site was resistant to cyanide as in E. coli oxidase while the quinol oxidation site was more sensitive to antimycin A and quinolone inhibitors. Spectroscopic analysis showed the presence of the haem b(595)-d binuclear centre but the sequence analysis indicates that cyanobacterial cytochrome bd is structurally related to cyanide-insensitive oxidase (CioAB), which does not show typical spectral changes upon reduction and ligand binding. Our data indicate that cyanobacterial cytochrome bd has unique enzymatic and structural properties and we hope that our findings will help our understanding the role and properties of CydAB and CioAB quinol oxidases in other bacterial species. PMID- 19124294 TI - Smoking among Vietnamese health professionals: knowledge, beliefs, attitudes, and health care practice. AB - The study aimed to investigate smoking patterns and compare knowledge, beliefs, and attitudes, as well as explore predictors of smoking status among Vietnamese health professionals. A global survey questionnaire on tobacco use among health professionals by the World Health Organization and the Centers for Disease Control and Prevention was adapted for data collection. Data from 2151 health workers from the 3 largest hospitals--each of which is located in South, Central, and North Vietnam, respectively--were collected using quantitative methods. Both descriptive and inferential statistics were used in the data analysis. Smoking prevalence among Vietnamese health professionals is 13.4%, dominant among male health professionals compared with female counterparts (35.6% vs 1.8%), and significantly varies by regions. Physicians and dentists display a greater smoking proportion than nurses (23.0%, 10.5%, and 7.7%, respectively). The findings highlight the importance of improving and promoting beliefs of health professionals about being role models for their patients by not smoking. Special attention should be given to the following slogan: "Health professionals should act as nonsmoking role models for their patients and the public." This message should be incorporated into cigarette restriction regulations and policies at hospitals and recognized as one of the effective measures in cigarette control in the hospital context. PMID- 19124295 TI - Smoking among Vietnamese medical students: prevalence, costs, and predictors. AB - This cross-sectional study investigated smoking patterns and predictors among Vietnamese medical students. In total, 4720 medical students from 3 universities, each located in South, Central, and North Vietnam, were interviewed using an adapted Global Youth Tobacco Survey Questionnaire. Ideas on smoking behavior and tobacco control derived from group discussions with students and from unstructured interviews with student managers and university directors. Overall, the current smoking rate was 25.0% and dominant in men at 43.7%. Most started smoking when just entering university (18 +/- 3.3 years). Male students from the central region had the highest smoking rate (35.1%), whereas their northern counterparts had the greatest smoking magnitude: age of smoking initiation (18.6 +/- 3.5), number of cigarettes per day (4.4 +/- 4.5), number of smoking days per month (16.5 +/- 11.6), and proportion of smoking cost among total expenses per month (10.9% +/- 11.9%). Smoking tended to increase across academic years, being highest in years 5 to 6 (35.0%). In contrast, the practice of ever quitting and the intention of quitting tended to decline from years 1-2 to years 5-6 (from 82.2% to 71.5%, P < .05, and from 70.8% to 51.5%, P < .001, respectively). Positive attitudes toward smoking (odds ratio = 1.4, P < .05), negative beliefs on hazards of smoking (odds ratio = 1.7), and daily exposure to family smokers (odds ratio = 2.0, P < .05) and to social smokers (odds ratio = 4.5, P < .05) were main predictors of smoking. Qualitative results suggest that nonsmoking university regulations played a critical role in tobacco control among medical students. Nonsmoking regulations and penalties for students who smoke need to be formulated at medical universities. The message that medical students must make a nonsmoking role model for community should be systematically promoted. PMID- 19124296 TI - Exposure to secondhand smoke in Kanchanaburi demographic surveillance system, Thailand. AB - Using data from the Kanchanaburi demographic surveillance system in Thailand, this article documents that tobacco smoke affects 60% of the population. The main effect is through exposure to secondhand smoke. More than half of men are smokers compared with only one tenth of women. Most men tend to start smoking during their teenage years, whereas the majority of women start at later ages. The majority of households are exposed to secondhand smoke. The highest level of exposure is in rural areas. Smokers are most likely to be male and older, but those exposed to secondhand smoke tend to be female and younger. Exposure to secondhand smoke is more likely to occur in households with lower socioeconomic status. Logistic regression analysis supports 2 study hypotheses: that children and women are the most affected by secondhand smoke, and household factors are the most important factors affecting the exposure to secondhand smoke. PMID- 19124298 TI - Identifying types of domestic violence and its associated risk factors in a pregnant population in Kerman hospitals, Iran Republic. AB - The objective of this study was to estimate the prevalence of different kinds of physical and emotional violence in an Iranian pregnant population and to examine its associated risk factors. This cross-sectional study was done from March through July 2005 in the 4 main hospitals of Kerman, Iran, which had maternity units. In total, 416 out of 460 women who were asked to participate agreed to be interviewed, a 90.4% response rate. All respondents were interviewed privately during the first 48 hours after delivery. The mean age (+/- SD) was 28.0 +/- 5.6, and all were married. Most of the women were urban residents (89.2%), and the majority of them were multiparous (78.8%). Nearly 16% of mothers said the pregnancies were unintended. In total, 35% (95% confidence interval: 30%-40%) of women had experienced 1 or more episodes of emotional violence during the pregnancy inflicted by their husbands, and 106 women (25%; 95% confidence interval: 21%-30%) had experienced at least 1 episode of physical violence. The highest odds of domestic violence during pregnancy was associated with unintended pregnancies (odds ratio: 7.66; 95% confidence interval: 3.45-16.99) and multiparous pregnancies (odds ratio: 6.88; 95% confidence interval: 3.46-13.68). Considering the high prevalence of different types of domestic violence during pregnancy, it should be regarded as a priority for health policy experts in Kerman and possibly Iran. PMID- 19124297 TI - Indoor air pollution and respiratory function of children in Ashok Vihar, Delhi: an exposure-response study. AB - The objective of this study was to investigate the effects of indoor air pollution on respiratory function of children (aged 7-15 years). The study took place at Ashok Vihar, an urban locality in the northwest part of Delhi during the summer months of June and July 2004. The team did house visits. The questionnaire, administered at the house itself, asked about the history of smoking in the family, type of cooking fuel used, duration of cooking, ventilation and lighting at the cooking place, and other confounders. In total, 441 children (59% male, 41% female) between ages 7 and 15 years were considered for the study, and a detailed profile was collected. Clinical examination with special reference to respiratory system was done. Pulmonary function tests/peak expiratory flow rates of each child were measured. Indoor air pollutant (suspended particulate matter, SO(2), NO(2)) was measured, and the effect of these pollutants on the children's respiratory function was analyzed. The respiratory health profile suggests that children had cough, sputum production, shortness of breath, wheezing, common cold, and throat congestion. Indoor SO(2) , NO(2), and suspended particulate matter levels were high in houses where there was a family history of smoking. SO(2) level was significantly high according to occupancy per room. NO(2) and suspended particulate matter levels were significantly high in houses where children had respiratory problems. It is concluded that indoor air pollution had an association with respiratory function of children. PMID- 19124299 TI - Larvivorous potential of the guppy, Poecilia reticulata, in anopheline mosquito control in riverbed pools below the Kotmale dam, Sri Lanka. AB - The larvivorous potential of Poecilia reticulata was studied in the laboratory and in the field. In the laboratory, the number of anopheline larvae consumed per fish per day and the number of larvae in fecal matter per fish were determined. In the field study, 29 of 60 selected riverbed pools in Kotmale oya, below the Kotmale dam, were stocked with P reticulata, whereas the rest served as controls. Anopheline larval surveys were carried out starting from 1 day prior to stocking of fish and on selected days subsequently. P. reticulata consumed an average of 117 +/- 32.33 larvae per fish per day. Fecal matter of P. reticulata contained an average of 2.7 +/- 2.68 larvae per fish . There was a significant reduction in the number of pools positive for anopheline larvae (P < .001) after stocking fish and the number of larvae per pool (P < .001) and per 100 dips (P < .001) as compared with controls. PMID- 19124300 TI - Climate change and the transmission of vector-borne diseases: a review. AB - This article reviews studies examining the relationship between climate variability and the transmission of vector- and rodent-borne diseases, including malaria, dengue fever, Ross River virus infection, and hemorrhagic fever with renal syndrome. The review has evaluated their study designs, statistical analysis methods, usage of meteorological variables, and results of those studies. The authors found that the limitations of analytical methods exist in most of the articles. Besides climatic variables, few of them have included other factors that can affect the transmission of vector-borne disease (eg, socioeconomic status). In addition, the quantitative relationship between climate and vector-borne diseases is inconsistent. Further research should be conducted among different populations with various climatic/ecological regions by using appropriate statistical models. PMID- 19124301 TI - Mice, pigs and public health. PMID- 19124302 TI - The reliability of dietary and lifestyle information obtained from spouses in an elderly chinese population. AB - In many health studies of the elderly population, the subjects have cognitive or linguistic impairments, so data need to be collected from surrogates. This study compares dietary and lifestyle information reported by elderly Chinese with those provided by their spouses. Community couples 60 years and older were recruited to participate in an interview. One person from each couple was randomly chosen as the index person. Characteristics concerning the index person were then solicited from that person and separately from his or her spouse using validated questionnaires. For the 128 food items considered, the mean kappa was 0.73 for both frequency (SD 0.18) and amount (SD 0.22) of intake, and more than 70% of the couples had kappa statistics exceeding 0.61. Food items exhibiting high agreement between the spouses include rice, apples, tomatoes, and pork chops. The proportion of perfect agreement was higher than 80% for physical activity, smoking, and tea drinking behaviors. In conclusion, the spouse can serve as a proxy to provide reliable information when his or her partner is unavailable. PMID- 19124303 TI - Community smoking behavior in Changqiao, Shanghai. AB - China has observed increasing tobacco use in the past decade. Only a few studies describing smoking behavior were reported at the community level in China. The present research aimed to obtain baseline data on tobacco use and factors that influence smoking behaviors in a local community in Shanghai, China. A total of 2100 residents in Changqiao, Shanghai, between ages 13 and 84 years were surveyed using a multistage proportional random sampling design. On the basis of that, a subsampling was conducted, and 1500 residents were randomly selected to avoid a cluster effect. There were 28.3% ever smokers (53.3% for men and 2.3% for women) and 23.5% current smokers (44.2% for men and 1.9% for women). Men ages 40 to 49 years smoked the most (more than 17 cigarettes per day on average). Age, education, and marital status were significant predictors of current smoking among men. Only 13.4% of current smokers were willing to quit. The prevalence of secondhand smoke exposure was 30.4% among nonsmokers. Smoking and secondhand smoking are serious problems in Changqiao, Shanghai. Comprehensive and intensive interventions should be implemented to motivate quitting and reduce secondhand smoke exposure. PMID- 19124304 TI - Early detection and prevention of domestic violence using the Women Abuse Screening Tool (WAST) in primary health care clinics in Malaysia. AB - Despite being an emergent major public health problem, little research has been done on domestic violence from the perspectives of early detection and prevention. Thus, this cross-sectional study was conducted to identify domestic violence among female adult patients attending health centers at the primary care level and to determine the relationship between social correlates of adult patients and domestic violence screening and subsequent help/health-seeking behavior if abused. Face-to-face interviews were conducted with 710 female adult patients from 8 health centers in Selangor who matched the inclusion criteria and consented to participate in the study, using a structured questionnaire that included adaptation of a validated 8-item Women Abuse Screening Tool (WAST). Statistical tests showed significant differences in ethnicity, income, and education between those screened positive and those screened negative for domestic violence. Of the participants, 92.4% reported that during consultations, doctors had never asked them whether they were abused by their husband/partner. Yet, 67.3% said they would voluntarily tell the doctor if they were abused by their husband/partner. The findings indicate that primary care has an important role in identifying domestic violence by applying the WAST screening tool, or an appropriate adaptation, with women patients during routine visits to the various health centers. Such assessment for abuse could be secondary prevention for the abused women, but more important, it will serve as primary prevention for nonabused women. This approach not only will complement the existing 1-stop crisis center policy by the Ministry of Health that copes with crisis intervention but also will spearhead efforts toward prevention of domestic violence in Malaysia. PMID- 19124305 TI - Reproductive health awareness among adolescent girls in rural Bangladesh. AB - This article presents the status of rural Bangladeshi adolescent girls' awareness about reproductive health. Analysis of data revealed that a sizable proportion of adolescent girls had incorrect knowledge or misconceptions about the fertile period, reproduction, sexually transmitted diseases, and HIV/AIDS. Age, education either of adolescents or their mothers, residence, and exposure to mass media were the significant predictors of adolescent girls' knowledge about reproductive health. Strong efforts are needed to improve awareness and to clarify misconceptions about reproductive health. Improved access to mass media and education could improve rural Bangladeshi adolescent girls' awareness about reproductive health. PMID- 19124306 TI - Treatment-seeking behavior and treatment practices of lymphatic filariasis patients with lymphoedema in the Colombo district, Sri Lanka. AB - This cross-sectional, descriptive study describes the treatment-seeking behavior of 413 lymphoedema patients attending 2 filariasis clinics in the Colombo district, Sri Lanka. A pretested, interviewer-administered questionnaire obtained information regarding sources and types of treatment taken, time taken for diagnosis, and details regarding diethylcarbamazine citrate (DEC) treatment. There was a mean delay of 2.37 years (SD 1.37) in diagnosing filariasis after the first appearance of limb swelling. General practitioners were the most frequent first-contact health care providers and the most visited source overall, followed by government hospitals and Ayurvedic practitioners. Approximately 95% of patients were on DEC treatment ranging from 10 days to 43 years (mean 2.5 years SD +/- 1.1). Sixty-one percent of patients reported always having taken the recommended DEC course. Nonsteroidal anti-inflammatory drugs, diuretics, and antibiotics were liberally prescribed. Approximately 97% had sought treatment from a medical practitioner for an acute adenolymphangitis attack. Despite the area being endemic for filariasis, there was a delay in treatment and inappropriate use of DEC in patients with chronic filarial lymphoedema. PMID- 19124307 TI - Sharps injury and body fluid exposure among health care workers in an Australian tertiary hospital. AB - To examine sharps injury and body fluid exposure among health care workers, a descriptive epidemiological study was conducted in a 1000-bed tertiary hospital between 2000 and 2003 using surveillance data of all reported sharps injuries and body fluid exposures. A total of 640 sharps injuries and body fluid exposures were reported from hospital and nonhospital staff, although no seroconversions to HIV, hepatitis B virus, or hepatitis C virus were observed during the study period. Nurses reported 47% of sharps injuries and 68% of body fluid exposures, medical staff reported 38% and 16%, and other nonmedical staff notified 5% and 4%, respectively, while nonhospital staff reported the rest. Hollow-bore needles accounted for 56% of sharps injuries, while 11% of the incidents were sustained during recapping and inappropriate disposal. Further research into Australian work practices, disposal systems, education strategies, and the use of safety sharps should be emphasized to implement strategies to reduce work-related injuries among health care workers. PMID- 19124308 TI - Deaths due to external causes among adolescents in Hong Kong. AB - The authors compared adolescent mortalities due to external causes in two 5-year periods (from 1986-1990 and 1996-2000) in Hong Kong. Leading external causes are traffic accidents and suicide. The mortality rate is comparatively low and has decreased significantly over time. Suicide deaths are, however, on the increase. Monitoring the changing trend is useful in setting public health priorities. PMID- 19124309 TI - Association between pregnancy induced hypertension and low birth weight; a population based case-control study. AB - The purpose of this study was to investigate the association between pregnancy induced hypertension and low birth weight. A population-based case control study was conducted. Antenatal mothers who attended the government health centers in the district of Kuala Muda, Kedah, Malaysia from June 2003 to May 2004 were recruited. Cases were 312 mothers who delivered low birth weight babies, and controls were 312 mothers who delivered normal birth weight babies. Face-to-face interviews using a structured questionnaire and a review of medical records were carried out. After controlling for important confounders such as gestational age at delivery, maternal age, ethnicity, education, parity, and previous history of abortion, pregnancy-induced hypertension was found to be an independent risk factor (adjusted odds ratio = 5.06; 95% confidence interval: 2.63, 9.71) for low birth weight. There was a significant association of pregnancy-induced hypertension with low birth weight. Women who delivered low birth weight babies were 5 times more likely to have had pregnancy-induced hypertension. PMID- 19124310 TI - Urban-rural disparity of overweight, hypertension, undiagnosed hypertension, and untreated hypertension in China. AB - The objective of this study was to examine the urban-rural variations in the prevalence of overweight, hypertension, undiagnosed hypertension, and untreated hypertension among adults. The China Health and Nutrition Survey was used to study adults 25 to 65 years old. Logistic regression was used to obtain the odds ratios (ORs) after adjusting for demographic, socioeconomic, and lifestyle variables. Urban adults have a higher probability of being overweight (OR = 1.18, P < .01) and having hypertension (OR = 1.19, P < .1). After further controlling for lifestyle variables, the difference is not significant. Urban hypertensive adults are less likely to be undiagnosed (OR = 0.54, P < .001) and untreated (OR = 0.53, P < .001), even after controlling for socioeconomic and lifestyle variables, suggesting that there are other reasons for the urban-rural disparity of undiagnosed and untreated hypertension. This finding suggests that modifying lifestyles is critical in preventing urban adults from being overweight. Interventions targeting rural adults should promote the awareness and treatment of hypertension. PMID- 19124311 TI - A journal for public health issues in the Asia Pacific: APJPH. PMID- 19124312 TI - Cigarette smoking status and smoking cessation counseling of Chinese physicians in Wuhan, Hubei province. AB - Among the 347 physicians surveyed, 58% of the male physicians and 18.8% of the female physicians were current cigarette smokers; 54.4% of the male and 70.4% of the female physicians often or always provided smoking cessation counseling for patients; 37.5% of the physicians thought that for a Chinese smoker, cigarette smoking served as a social lubricant; 31.5% thought it a habit; 21.7% thought it a stress reliever; and 9.2% thought it a social status symbol. The following 5 variables were significantly associated with physicians' smoking cessation counseling frequency: their smoking status, perceived success in their past counseling, perceived influence, perceived exemplary role, and perceived responsibility. To increase physicians' smoking cessation counseling, the Chinese Ministry of Health would need to discourage physicians to smoke and appeal to their sense of responsibility to help patients quit smoking. PMID- 19124314 TI - Smoking among young urban Malaysian women and its risk factors. AB - This study was conducted to measure the prevalence of cigarette smoking and to determine the individual, family, and environmental factors associated with smoking among young urban women. A cross-sectional study through self administered questionnaire was conducted on female students enrolled in private higher learning institutions in Kuala Lumpur and Selangor, Malaysia, between July and October 2005. Analysis on 408 respondents showed that current smoker prevalence rate was 18.6%. Adjusted analyses showed significant association between smoking and individual factors, which are the importance of slim image, average monthly allowance, and car ownership. For family factors, analyses showed significant association between smoking and parental marital status and smoking status of male siblings. Strong associations were seen between female smoking and environmental factors, such as having more smoker friends, having smokers as best friends, keeping cigarette-brand items, being offered free cigarette, and perceiving female smoking as normal. The identified risk factors could be used to develop more effective prevention programs to overcome smoking among young urban women. PMID- 19124313 TI - Patterns of smoking among adolescents in Malaysia and Thailand: findings from the International Tobacco Control Southeast Asia survey. AB - At present, 70% of the world's 1.1 billion smokers are in developing countries, with over 50% in Asia alone. The current study examined patterns of youth smoking in Thailand and Malaysia. Respondents were 2002 youths between the ages of 13 and 17 from Thailand (n = 1000) and Malaysia (n = 1002). Respondents were selected using a multistage cluster sampling design and surveyed between January 2005 and March 2005. Approximately 3% of youth between the ages of 13 and 17 were current smokers, with an additional 10% to 12% reporting experimental smoking. Males were between 7 and 15 times more likely to report smoking behavior than females. Less than 1% of females respondents in either country met the criteria for current smoking, and less than 5% met the criteria for experimental smoking. In contrast, more than 50% Thai males and approximately one-third of Malaysian males aged 17 met the criteria for either experimental or current smoking. PMID- 19124315 TI - Factors contributing to medication noncompliance of newly diagnosed smear positive pulmonary tuberculosis patients in the district of Colombo, Sri Lanka. AB - Medication noncompliance hinders effective tuberculosis control. This descriptive study investigates the factors contributing to medication noncompliance among new patients with smear-positive pulmonary tuberculosis on treatment at government health institutions in Colombo, Sri Lanka. In a cohort of patients aged > or =15 years (n = 326), 23% were found to be noncompliers (n = 74) on follow-up. The median age of noncompliers (50 years) was significantly higher than the compliers (45 years). In multivariate logistic regression analysis, factors associated with noncompliance are as follows: being a male, living alone or with extended family, experiencing side effects to medication, perceiving nonsusceptibility to adverse effects of illness, and perceiving no benefit in regular treatment. The participants of a focus group discussion on service factors opined that the reception at treatment facilities and the interaction with certain categories of staff were poor. Noncompliance is related to a multiplicity of factors involving patients and healthcare services. PMID- 19124316 TI - Epidemiology of insomnia in Malaysian adults: a community-based survey in 4 urban areas. AB - This study aimed to determine the prevalence and pattern of insomnia in a Malaysian population aged 30 to 70 years. The sample consisted of 1611 subjects, recruited by stratified random sampling and interviewed using a semistructured questionnaire conducted in 2004. This was a community-based survey in 4 Malaysian states. The prevalence of insomnia symptom was 33.8%, and 12.2% of the subjects had chronic insomnia. Insomnia was more common among elderly; those who were separated, divorced, or widowed; and those who smoked at bedtime. Subjects with insomnia had a higher prevalence of feeling depressed (12.7), loss of concentration (19.1%), exhaustion (17.2%), poor memory (9.2%), decreased work productivity (6.4%), and perceived poor health status (40.9%; all, P < .05). A total of 22.2% of those with insomnia had excessive daytime sleepiness based on their Epworth Sleepiness Score (P = <.001). Those with insomnia used more sedatives (9.9%) compared with those without insomnia (2.6%; P < .001). About one third of the population had insomnia associated with impaired daily function. This study concluded that insomnia is common in Malaysian adult population, and it has significant impact on psychological well being and daily functioning. PMID- 19124317 TI - Problems related to menstruation and their effect on daily routine of students of a medical college in Delhi, India. AB - Menstrual problems are common among young girls. These deserve careful evaluation as uncorrected menstrual problems may adversely affect the daily routine and quality of life. The aim of this article is to study the prevalence and the effect of menstrual disorders on daily routine among unmarried undergraduate medical students and their treatment-seeking behavior. Of 276 undergraduate girl students, 112 were sampled by stratified random sampling. All the consenting participants were given a pretested semistructured questionnaire to collect their responses by personal interviews. The data collected were analyzed. Premenstrual syndrome (67%) and dysmenorrhea (33%) were perceived by the study subjects as the most distressing problems associated with menstruation. The most common effect of menstrual problems on daily routine reported by the study subjects was in the form of prolonged resting hours (54%) followed by inability to study (50%). More than half (52%) of the subjects discussed their problems with their mother, and 60% of the study subjects were opted for allopathic treatment for their menstrual problems. PMID- 19124318 TI - Current developments in genomics and personalized health care: impact on public health. AB - The knowledge gained from the characterization of genomes, especially the human genome, holds considerable potential for the development of new health care innovations for prevention, diagnosis, and management of many diseases in the coming decade. However, owing to the presence of highly complex scientific, economic, social, and ethical issues associated with this field, societies will need to be better prepared for the era of postgenomics and its consequences. It is important to ensure that the benefits of genomics are distributed fairly among all the countries of the world and that the well-tried and more conventional approaches to medical research and practice are not neglected while the medical potential of genomics is being explored. In this report, the author focuses mainly on human genomics, its applications, development of related technologies and issues related to the dissemination of knowledge derived from genome information, and finally, their impact on global health care. PMID- 19124320 TI - Researching women's health in Asia: are we doing enough? PMID- 19124319 TI - Risk factors for fragility fracture in Seremban district, Malaysia: a comparison of patients with fragility fracture in the orthopedic ward versus those in the outpatient department. AB - Osteoporosis is a silent disease and becomes clinically significant in the presence of fragility fracture. Identifying risk factors that are associated with osteoporosis in the community is important in reducing the incidence of fragility fracture. The aim of this study is to identify risk factors associated with fragility fracture in the Seremban District of Malaysia. This is a population comparison study between orthopedic ward patients and outpatients attending a community health clinic for 6 months. Epidemiological data and the possible risk factors for osteoporosis were collected by direct interview. This study demonstrates that advancing age, low body weight, smoking, lack of regular exercise, low consumption of calcium containing foods, and using bone depleting drugs (steroids, thyroid hormone, and frusemides) are major risk factors for fragility fracture. Most of these risk factors are modifiable through effective lifestyle intervention. PMID- 19124321 TI - The evolving evidence base for child protection in Chinese societies. AB - Child maltreatment is a substantial public health problem worldwide. Although extensively studied in Western countries, until recently little systematic research had been published about the situation in the world's most populous nation and ethnic diaspora. In this review, we examine trends from community based research with Chinese young people and parents in mainland China, Hong Kong SAR, Taiwan, Singapore, and Malaysia. It is clear that many Chinese adolescents experience a substantial burden from various forms of maltreatment and the psychological and behavioral correlates are similar to those found in other cultures. However, the research reveals a large gap between this reality and Chinese adults' perceptions about emotional, physical, and sexual abuse. Comprehensive awareness programs are needed to close this information gap and thereby mobilize support for prevention and care initiatives. PMID- 19124322 TI - Evaluation of a health education intervention on knowledge and attitudes of food handlers working in a medical college in Delhi, India. AB - The present study assessed change in knowledge, attitudes, and self-reported hand washing practices of 136 food handlers 3 months after providing them health education using posters and interactive sessions using a flip chart. There was a significant change in the proportion of study participants who named food borne or waterborne diseases, including gastroenteritis (79.4% to 91.9%), worm infestation (39.0% to 55.9%), cholera (16.2% to 42.7%), and typhoid (9.6% to 28.7%). Significant increase in knowledge about hand hygiene measures, namely, washing hands before handling food (23.5% to 65.4%) and keeping nails cut and clean (8.1% to 57.4%), was observed. Baseline self-reported hand-washing practices revealed low figures for washing hands after micturition (82.4%) and smoking (52.8%) and consistent use of soap at the workplace (24.3%) and after micturition (14.0%), which improved after health education but not to the desired extent. Findings highlight the importance of providing health education in food and personal hygiene to food handlers and incorporating the same in existing guidelines for food establishments laid down by civic agencies in Delhi and elsewhere. PMID- 19124323 TI - Cooperation, integration, and long-term commitment: what Solomon Islanders and development workers say about health sector aid. AB - INTRODUCTION: The Solomon Islands is experiencing instability and insecurity and also a concomitant increase in aid. This article aims to address the need for theoretical coordination frameworks to be further informed by the actual experiences, requirements, and views of the recipients of aid. METHODS: Qualitative research techniques were used to better understand governmental and nongovernmental leaders' views of health sector aid in the Solomon Islands. Data were collected using previously published literature, government and nongovernmental documents, and in-person interviews. RESULTS: Two key themes emerged from the interviews: the need for coordination and integration of aid and the need for this integration to occur over the long-term. These themes are presented using quotations from key informants. CONCLUSION: Themes and quotations arising from the analyses may assist in understanding theoretical frameworks for coordination, particularly in postconflict states. Future needs regarding mechanisms of collaboration in the Solomons are also discussed. PMID- 19124324 TI - Orgasmic dysfunction among women at a primary care setting in Malaysia. AB - AIM: The aim of this study was to investigate the prevalence of orgasmic dysfunction and the potential risk factors that may be associated with orgasmic dysfunction among women at a primary care setting in Malaysia. METHODS: A validated questionnaire for sexual function was used to assess orgasmic function. A total of 230 married women aged 18 to 70 years participated in this study. Their sociodemographic and marital profiles were compared between those who had orgasmic dysfunction and those who did not, and the risk factors were examined. RESULTS: The prevalence of orgasmic dysfunction in the primary care population was 51.9%. Women with orgasmic dysfunction were found to be significantly higher in the following groups: age >45 years, being non-Malay, having lower academic status, married longer, having more children, married to an older husband, and being at menopausal state. CONCLUSION: Women with infrequent sexual intercourse are less likely to be orgasmic (odds ratio = 0.29, 95% confidence interval = 0.11 0.74). PMID- 19124325 TI - The economic evaluation of a health screening program on congenital heart disease for school children in Taichung, Taiwan. AB - OBJECTIVE: The objective of this study was the economic evaluation of screening school children for congenital heart disease (CHD). METHODS: The sample consisted of all the 45,725 students in grades 1 and 7 from Taichung County, Taiwan. STATISTICA, version 7.1, was used to determine descriptive statistics and Student's t tests on students' height, weight, and body mass index (BMI). RESULTS: The total cost of CHD screening was NT$4,863,866 (NT$32 = US$1), and the average cost was NT$106.37 per child. The rate of finding new asymptomatic students was 2.9174%, and the cost of finding 1 asymptomatic student was NT$3646. Also, a cost-effective analysis showed NT$165.7 per year of life saved. CONCLUSIONS: Twelve Student's t tests were conducted, and it found that students' height, weight, and BMI was not associated with asymptomatic students. A goodness of-fit chi(2) found that females have a higher rate of asymptomatic CHD. PMID- 19124326 TI - The influence of low blood lead concentrations on the cognitive and physical development of primary school children in Malaysia. AB - This study aimed to determine the relationship between blood lead (BPb) concentrations and cognitive and physical development in school children. A total of 169 urban children and 100 industrial children of Malay ethnicity, in the age range of 6(1/2) to 8(1/2) years, were selected. BPb was determined using GF atomic absorption spectrophotometer. The mean cognitive score (102.55) of the children from the industrial area was significantly higher than that of the urban children (95.09; P < .001). However, no significant differences were found in the BPb levels between the 2 groups (industrial, 3.75 microg/dL; urban, 3.56 microg/dL). There was significant inverse correlation between BPb and cognitive scores for all children (P < .05). The cognitive scores for all children were influenced by BPb after adjustments (P < .05). The urban children had significantly better Weight for Height and Left Arm Circumference values than those from industrial area. There was no significant correlation between BPb and the anthropometric measurements. In conclusion, low BPb influenced the cognitive development, whereas physical development was not affected. PMID- 19124327 TI - Increased fatalities and cost of traumatic injuries in elderly pedestrians in Hawaii: a challenge for prevention and outreach. AB - This study was carried out to evaluate and quantify risk factors, mechanisms, and cost of traumatic injuries in Hawaii's elderly and to identify potential preventive strategies. A retrospective review of a prospective database of patients admitted to the only Trauma Center in the Pacific Basin, between January 2000 and December 2001, was conducted. Of 2634 trauma admissions, 11% were >or=65 years of age. Mechanisms of injury included falls, motor vehicle crashes (MVCs), pedestrians hit by automobiles, and miscellaneous causes. The incidence of elderly pedestrians hit by automobiles in Hawaii is higher than previously reported. Hospital mortality rate was highest for the pedestrian hit group, followed by falls, and then MVCs. The pedestrian hit group consumed the largest quantity of resources and MVCs the least. Given the high mortality rate and associated resource consumption in the pedestrian hit group, it would be appropriate to give priority to this group while developing preventive measures for this age group. PMID- 19124328 TI - Prevalence of rubella susceptibility among pregnant mothers in a community-based antenatal clinic in Malaysia: a cross-sectional study. AB - INTRODUCTION: routine rubella antibody screening is not done for antenatal mothers in community health clinics in Malaysia. However, congenital rubella syndrome has persisted with its associated health burden. OBJECTIVES: to determine the prevalence of rubella susceptibility among pregnant mothers and its associated risk factors. METHODOLOGY: a cross-sectional study was carried out in the Petaling district, Selangor, Malaysia, where 500 pregnant mothers were recruited, and face-to-face interviews were conducted. Rubella IgG tests were performed. RESULTS: the prevalence of rubella susceptibility among pregnant mothers was 11.4%. Using logistic regression, a history of not having received rubella vaccination or having unknown rubella vaccination status was found to be a significant predictor for mothers to be rubella susceptible (odds ratio = 2.691; 95% confidence interval = 1.539-4.207). CONCLUSIONS: routine rubella IgG screening tests need to be offered to all antenatal mothers in view of the high prevalence found. PMID- 19124329 TI - Prevalence of anemia and its determinants among nonpregnant and pregnant women in India. AB - This study investigates the severity and distribution of anemia among nonpregnant and pregnant women aged 15 to 49 years in urban and rural sectors of 26 states in India and its association with certain economic and biosocial factors. A national survey was conducted to collect data on hemoglobin, height, weight, and certain economic and biosocial factors of 72 660 nonpregnant and 5619 pregnant women. Nonpregnant women aged less than 25 years are most affected by anemia. Women's education and standard of living in the households have a vital role in reducing anemia. Urban and well-nourished women also suffer less from anemia. The severity of anemia is higher among pregnant than nonpregnant women. Efforts must be made to educate women and enhance their level of economic status so that the prevalence of anemia can be reduced substantially. PMID- 19124330 TI - Depressive symptoms in middle-aged women in peninsular Malaysia. AB - A household survey was carried out in Peninsular Malaysia to determine the prevalence rate of depressive symptoms in middle-aged women and to explore its associated factors. Women aged 45 to 60 years were assessed on sociodemographic profiles, menopausal status, depressive symptoms, marital dissatisfaction, and coping strategies. A total of 3934 women participated, and the mean age was 51 years. The prevalence rate of depressive symptoms was 54.2%. Of the married women, 38.4% had poor to very severe problems in their marriage. Depressive symptoms were significantly associated with marital status (P < .001), ethnicity (P < .001), educational levels (P < .001), occupational status ( P < .001), domicile (P < .001), and menopausal status ( P < .05). Depressive symptoms were correlated with marital dissatisfaction scores. Women who consulted professionals, shared with relatives, kept problems to themselves, or consulted parents/siblings had higher mean scores on depressive symptoms compared with those who did not use these coping strategies. PMID- 19124331 TI - Factors associated with inconsistent condom use among female sex workers in Nha Trang, Vietnam. AB - This study examined consistent condom use with clients, self-reported sexually transmitted infections (STIs), and associated factors among Vietnamese female sex workers (FSWs). Data were collected through a cross-sectional survey among 192 FSWs in Nha Trang city, in 2005. The rate of consistent condom use with casual clients, regular clients, and nonpaying partners were 59.3%, 38.3%, and 5.3%, respectively. About 5.7% of the FSWs admitted that they are intravenous drug users, and 66.7% of the sample had symptoms of STIs in the past 12 months. Indirect FSW, being drug users, having many clients per week, not watching television on a regular basis, those who thought that they themselves are at extremely high risk of having HIV, and those have never been tested for HIV have been found to be associated with inconsistent condom use. Our results suggest the need to improve STI control in FSWs, further promotion of condom use and information, education, and communication among FSWs. PMID- 19124332 TI - Child immunization coverage in rural hard-to-reach Haor areas of Bangladesh: possible alternative strategies. AB - This article assessed the status of childhood vaccination coverage and the possibility of using selected alternative vaccination strategies in rural hard-to reach haor (low lying) areas of Bangladesh. Data were collected through survey, in-depth interviews, group discussion, and observations of vaccination sessions. Complete immunization coverage among 12- to 23-month-old children was found to be significantly lower in study areas when compared with the national coverage levels. The study identified reasons for low complete immunization coverage in hard-to-reach areas, including irregular/cancelled extended program on immunization (EPI) sessions, less time spent in EPI spots by field staff, and absence of any alternative strategy for remote areas. The findings indicated that the existing service delivery strategy is not sufficient to improve immunization coverage in hard-to-reach areas. However, most of the strategies assessed are considered possible to implement by health care providers in hard-to-reach areas. The study suggested that before implementing alternative strategies in hard-to reach areas, feasibility and effectiveness of the possible strategies need to be tested to identify evidence-based strategies. PMID- 19124333 TI - Assessment of mental health status of senior secondary school children in Delhi. AB - BACKGROUND: Schoolchildren undergo rapid mental, emotional, and social changes and are susceptible to psychiatric disorders. OBJECTIVES: This study aimed to assess the mental health status and some determinants in senior secondary school children in Delhi. METHODOLOGY: A cross-sectional study was carried out on a sample of 458 children, selected by 2-stage sampling. Goldberg's General Health Questionnaire containing 60 items (GHQ-60) was used for assessment of mental health. RESULTS: GHQ was administered to 239 boys and 219 girls in the age group of 15 to 20 years. Of 458 students, 113 (24.7%) had a score>or=16, the cutoff score, indicating mental morbidity among them. Among boys and girls, 28.5% and 20.5%, respectively, had some component of mental morbidity. Morbidity was significantly higher in children hailing from nuclear families and among children who had either failed or those who had scored highest in the class. CONCLUSION: Mental health needs of schoolchildren need to be addressed by the school health services. PMID- 19124334 TI - Sentinel surveillance of soil-transmitted helminthiasis in selected local government units in the Philippines. AB - This study describes baseline prevalence and intensity of soil-transmitted helminth infections as well as baseline anthropometric and school performance data among public elementary school children in 6 sentinel provinces in the Philippines. Stratified cluster sampling was used to select 6 provinces, where grade 3 elementary school pupils were surveyed. Secondary anthropometric data and achievement test results of the immediate past academic year were examined. Overall cumulative prevalence and proportion of heavy intensity infections for the 6 selected provinces were 54.0% and 23.1%, respectively. These recent findings further support the need for mass treatment to be given at least twice a year. The findings of the study also demonstrate the relationship that exists between worm burden and nutritional status. Strategies focusing on mass treatment integration, environmental sanitation, personal hygiene, and health education should be developed to control soil-transmitted helminth infections and their detrimental effects. PMID- 19124335 TI - Severity of visual impairment and depression among elderly Malaysians. AB - OBJECTIVE: This study aimed at evaluating the relationship between severity of visual impairment and depression among elderly Malaysians attending an eye clinic. METHODS: This was a cross-sectional study. The study population included patients, aged>or=60 years who attended the Eye Clinic in University Malaya Medical Centre. Exposure measurement was based on ophthalmologic examinations by an ophthalmologist. Data on outcome were measured using the Geriatric Depression Scale. RESULTS: After adjusting for important confounders, severity of visual impairment either having low vision or blind were independent risk factors of depression. The odds of developing depression among elderly with low vision were 2 times more than those with normal vision, and elderly who were blind had almost 5 times the odds to be depressed compared with those having normal vision. CONCLUSION: Findings from this study suggest a positive relationship between the severity of visual impairment and depression among elderly Malaysians. PMID- 19124336 TI - Chronic disease risk factors in rural Australia: results from the Greater Green Triangle risk factor surveys. AB - The aim of this article was to assess the level and prevalence of major chronic disease risk factors among rural adults. Two cross-sectional surveys were carried out in 2004 and 2005 in the southeast of South Australia and the southwest of Victoria. Altogether 891 randomly selected persons aged 25 to 74 years participated in the studies. Surveys included a self-administered questionnaire, physical measurements, and a venous blood specimen for lipid analyses. Two thirds of participants had cholesterol levels>or=5.0 mmol/L. The prevalence of high diastolic blood pressure (>or=90 mm Hg) was 22% for men and 10% for women in southeast of South Australia, and less than 10% for both sexes in southwest of Victoria. Two thirds of participants were overweight or obese (body mass index>or=25 kg/m2). About 15% of men and slightly less women were daily smokers. The abnormal risk factor levels underline the need for targeted prevention activities in the Greater Green Triangle region. Continuing surveillance of levels and patterns of risk factors is fundamentally important for planning and evaluating preventive activities. PMID- 19124337 TI - Home smoking restrictions among Koreans in Seoul. AB - The prevalence and correlates of home smoking bans in the Republic of Korea were examined using population-based data from telephone interviews with 500 Seoul adult residents in 2002. Most (97%) respondents indicated that they smoked, or that their spouse, other family member, or a regular friend smoked. Nearly all indicated that environmental tobacco smoke (ETS) is harmful. Only 19% of homes banned smoking, 65% allowed smoking anywhere, and 16% allowed smoking by special guests or in certain areas. The odds of having a full ban were significantly greater for men, married individuals, those less than 35 years or more than 50 years of age, non-smokers, individuals whose nonspouse family members did not smoke, and individuals with more sources of anti-ETS messages. Home smoking bans should be promoted by media campaigns and other tobacco control activities, because knowledge of the effects of ETS was already high. PMID- 19124338 TI - The stages of physical activity and exercise behavior: an integrated approach to the theory of planned behavior. AB - This research aims to verify whether it is possible to explain the health promoting behaviors based on sociodemographic characteristics by integrating the theory of planned behavior (TPB) proposed by Ajzen in 1988 and the transtheoretical model (TTM) proposed by Prochaska and DiClemente in 1983. In particular, the aim was to verify whether the variables of the TPB can properly distinguish the stages of change in exercise in the proposed integrated model and to figure out how attitude, subjective norm, perceived behavior control, influence, and intention-can explain the stages of change in exercise. Investigators who have taken previous training for the survey visited and interviewed 3658 people older than 30 years in the chosen town by multistage sampling method from July 27 to July 31, 2003. After the exclusion of inappropriate data out of 760 participants, only data from 584 participants were used for this research. chi2 test, t test, and 1-way analysis of variance were used to identify the difference between the distribution of the stages of change in exercise and the variables of the means. A discriminant analysis to verify the accuracy of the stages of change in exercise by means of the variables of the TPB and a path analysis to verify the fit of the integrated model were also used. The variables of the TPB were useful to satisfactorily distinguish and predict the stages of change in exercise. But to clarify the validity of this model, more diversified research should be conducted in the future, and the results must be accumulated. PMID- 19124339 TI - Estimating unit costs for dental service delivery in institutional and community based settings in southern Thailand. AB - In this cross-sectional study, the cost of different dental services was estimated and the unit costs of dental services for schoolchildren were compared between 2 settings: hospital-based and community-based mobile dental clinics. Heads of all departments in a selected community hospital were invited to attend 2 workshops to collect relevant data. Unit costs of different dental services varied from 41 to 2693 baht, with services falling into 4 unit cost groups: very high, high, moderate, and low. The very-high-unit-cost services included rehabilitative dental services. The high-unit-cost services covered removal of an impacted tooth, root canal treatment, and tooth-color fillings. The moderate-unit cost group included a wide range of other dental services, with screening and oral hygiene instruction in community-based dental clinics falling into the low unit-cost group. Generally, services provided in the community-based mobile clinic had lower unit costs than the same services provided in the hospital dental clinic. PMID- 19124340 TI - Public health policy and medical missions in the Philippines: the case of oral- facial clefting. AB - This article examines the critical role of medical missions as alternative or proxy health delivery agencies in developing countries like the Philippines. Why and how they exist, what they can and cannot do, how they interact with the for profit and public health sectors, and what challenges they face in the context of underdevelopment are analyzed by using a proposed structural- behavioral framework. We find that these missions can offer short-term benefits to specialized public health problems, such as oral-facial clefting, but depend on public and private partnerships and resources for long-term solutions. The article thus suggests that health care provision in developing countries should also be treated as a managerial issue. The challenge is for Third World governments to promote effective trisector collaborations, improve accessibility and adequacy of services, and support socially redistributive health policies. PMID- 19124341 TI - Socioeconomic determinants of age at first birth in rural areas of Bangladesh. AB - The age at first birth is very low under existing rural sociocultural settings in Bangladesh. This study examined the socioeconomic and cultural determinants of age at first birth. The study subjects were married women aged 15 to 29 years in 2 rural areas that were identified through a multistage sampling technique. To collect the relevant information, a semistructured interviewer schedule was applied to the eligible women. This study found that 72.8% women gave first birth at <20 years of age with mean age at first birth 18.74 years. Simple linear regression model explained 30.9% of variance in age at first live birth. Among socioeconomic and cultural determinants, family pressure explained the most significant variance. It is really difficult to reduce fertility in complex sociocultural settings in rural Bangladesh. However, the findings of this study may provide an answer to increase the age at first birth and hence to reduce the high fertility among these group of women. PMID- 19124342 TI - Clinicians' attitude on mass drug administration under the program to eliminate lymphatic filariasis: a qualitative study from Orissa, India. AB - The global program to eliminate lymphatic filariasis (LF) is currently based on mass drug administration (MDA) with annual single dose of antifilarials. In India, the MDA is implemented by primary health centers (PHC) of the district health system. The aim of the present study was to explore the attitudes of clinicians working at PHC level in three districts of Orissa, India. The data are obtained from interviews of 50 clinicians (35 from PHCs and 15 private). Only 11 PHC clinicians and 1 private clinician opined that the elimination of LF is possible through MDA of diethylcarbamazine (DEC); and 40% PHC and 87% private clinicians strongly felt that the elimination is not feasible through MDA and that the yearly single dose of DEC was not efficacious. The remaining clinicians were unable to comment on the feasibility and efficacy of MDA. The indifferent attitudes arise from lack of knowledge or erroneous beliefs. A good understanding of the program through reorientation and rigorous training should be emphasized to make the MDA successful. PMID- 19124343 TI - Editorial. A new respiratory journal. PMID- 19124344 TI - Analysis of exhaled breath condensate in respiratory medicine: methodological aspects and potential clinical applications. AB - Analysis of exhaled breath condensate (EBC) is a noninvasive method for studying the composition of airway lining fluid and has the potential for assessing lung inflammation. EBC is mainly formed by water vapor, but also contains aerosol particles in which several biomolecules including leukotrienes, 8-isoprostane, prostaglandins, hydrogen peroxide, nitric oxide-derived products, and hydrogen ions, have been detected in healthy subjects. Inflammatory mediators in EBC are detected in healthy subjects and some of them are elevated in patients with different lung diseases. Analysis of EBC is completely noninvasive, is particularly suitable for longitudinal studies, and is potentially useful for assessing the response to pharmacological therapy. Identification of selective profiles of biomarkers of lung diseases might also have a diagnostic value. However, EBC analysis currently has important limitations. The lack of standardized procedures for EBC analysis and validation of some analytical techniques makes it difficult comparison of results from different laboratories. Analysis of EBC is currently more useful for relative measures than for quantitative assessment of inflammatory mediators. Reference analytical techniques are required to provide definitive evidence for the presence of some inflammatory mediators in EBC and for their accurate quantitative assessment in this biological fluid. Several methodological issues need to be addressed before EBC analysis can be considered for clinical applications. However, further research in this area is warranted due to the relative lack of noninvasive methods for assessing lung inflammation. PMID- 19124345 TI - Rho kinase as a therapeutic target in the treatment of asthma and chronic obstructive pulmonary disease. AB - Asthma is a complex inflammatory disease of the airways involving reversible bronchoconstriction. Chronic obstructive pulmonary disease is typified by inflammation and airflow limitation that has an irreversible component. There is now substantial evidence that Rho kinase is involved in many of the pathways that contribute to the pathologies associated with these respiratory diseases including bronchoconstriction, airway inflammation, airway remodelling, neuromodulation and exacerbations due to respiratory tract viral infection. Indeed the Rho kinase inhibitor Y-27632 causes bronchodilatation and reduces pulmonary eosinophilia trafficking and airways hyperresponsiveness. Furthermore, accumulating evidence suggests that inhibition of Rho kinase could have a major beneficial impact on symptoms and disease progression in asthma and COPD by modulating several other systems and processes. Thus, the Rho kinase pathway may indeed be a worthwhile therapeutic target in the treatment of asthma and chronic obstructive pulmonary disease. PMID- 19124347 TI - Systemic consequences of COPD. AB - Chronic obstructive pulmonary disease (COPD) is a highly prevalent chronic lung condition, affecting approximately 10% of adults over the age of 40 years in the western world. Research over the past 10 years has shown that COPD is more than just a lung disorder; it affects other end-organs including the cardiovascular and the musculoskeletal systems, making it a multi-component, multi-system disease. COPD increases the risk for ischemic heart disease, stroke, osteoporosis, cachexia, and muscle weakness by two to threefold, independent of other factors such as smoking and age. The mechanisms by which COPD affects these end-organs, however, are unclear. In this paper, we review some of the common and serious extra-pulmonary manifestations of COPD and the potential mechanisms by which they can be linked with COPD. PMID- 19124346 TI - Safety of long-acting beta2-agonists in the treatment of asthma. AB - Several studies suggested an association between the regular use of beta2 agonists and asthma deaths. Whether this association represents adverse effects of beta -agonist use or is entirely due to disease severity is a matter of ongoing debate. Previous literature indicates that confounding by poor asthma control may explain the apparent deleterious effects of inhaled beta2-agonists. Tolerance to nonbronchodilator effects of beta2-agonists may account for the increase in reactivity to indirect bronchoconstrictor challenges and explain why some studies have demonstrated enhanced bronchoconstriction in patients with asthma after regular beta 2-agonist therapy. Nonetheless, the salmeterol multi centre asthma research trial (SMART) found more asthma deaths (13 vs 3) and life threatening asthma events (37 vs 22) in the salmeterol-treated asthmatic patients, although it was documented that among African-Americans, 5 times as many deaths and near-deaths from asthma occurred in those given salmeterol than in those given placebo, and among patients with asthma not using an inhaled corticosteroid (ICS) as a preventive (controller) medication, again more deaths and near-deaths from asthma occurred in those given salmeterol than in those given placebo. Only 38% of the African-Americans who participated in the study used an ICS. As a result of the findings from the SMART, FDA issued a public health advisory to highlight that long-acting beta2-agonists (LABAs) should not be the first medicine used to treat asthma. LABAs should be added to the asthma treatment plan only if other medicines, including the use of low-or-medium dose ICSs, do not control asthma. However, despite all of the concerns raised by the SMART, inhaled beta2-agonists remain the most effective bronchodilators available for the immediate relief of asthma symptoms and, as such, remain an important component of asthma management. Obviously, there are concerns about LABA treatment as monotherapy for asthma. Patients with asthma should be initiated and maintained on sufficiently high doses of ICSs and only patients whose asthma cannot be controlled should receive additional LABAs on a regular basis. PMID- 19124348 TI - Do we need new antibiotics for treating exacerbations of COPD? AB - Exacerbations may produce permanent impairment in lung function and health status in patients with COPD. Up to 70% of episodes have a bacterial etiology, being of mixed viral infection in some cases. The new, more active antibiotics have demonstrated better eradication of bacteria in the airways and, consequently, prolongation of the time to the next exacerbation. However, the ability of bacteria to develop resistance to the antibiotics currently used warrants novel research into new families of antimicrobials, and the adoption of new strategies such as the prevention of exacerbations, nebulized antibiotic treatment or the use of antibiotics in combination. PMID- 19124349 TI - The significance of tissue-imaging proteomics in respiratory therapies. PMID- 19124350 TI - Factors determining the quality of life of patients with COPD in primary care. AB - OBJECTIVES: To describe the health-related quality of life (HRQL) in a cohort of COPD patients recruited in primary care and identify the variables of the patients and the treatment associated with impaired HRQL. METHOD: Multicenter, observational study of patients with COPD recruited in Primary Care centers. Data regarding HRQL measured by the St. George's respiratory questionnaire (SGRQ) are presented. The total population was divided into two subgroups, high and Low SGRQ, using the median of the total SGRQ score. Univariate analysis and logistic regression analysis with calculation of adjusted odds ratios were used to investigate the factors significantly associated with a higher SGRQ score (poor HRQL). RESULTS: A total of 27 investigators included 222 valid patients with a mean age of 68.2 years (SD = 9.9) and a mean FEV1(%) of 49.2% (SD = 15.4%). The median total SGRQ score was 39.5 and patients with a high score had a longer evolution of COPD (p < 0.0001), more severe dyspnea (p < 0.0001) and a worse FEV1(%) (46.3% vs. 51.9%; p = 0.008). Factors independently associated with the total SGRQ score were cough and dyspnea, duration of COPD and treatment with inhaled steroids. CONCLUSION: : Patients with COPD controlled in primary care demonstrate an important impairment in HRQL, with chronic cough and dyspnea being associated with more impaired HRQL. Strategies aimed at modifying these factors should significantly improve the well-being of COPD patients. PMID- 19124351 TI - Smoking cessation treatment in a real-life setting: the Greek experience. AB - OBJECTIVES: The aim of this study was to estimate the clinical efficacy of counseling combined with currently used pharmacotherapy for smoking cessation (bupropion SR and nicotine replacement therapy, NRT) in actual clinical practice, and to identify predictors of successful abstinence at the end of therapy, as well as predictors of sustained abstinence in one year. METHODS: 895 smokers, self-motivated to quit, received bupropion SR for 7 or 19 weeks and/or nicotine replacement therapy (NRT) (nicotine patch) for 9 weeks in combination with individual behavioural therapy. An intensive program including repetitive visits and telephone contacts during treatment and one year's follow-up period was available for supporting and motivating smokers to prevent relapse. RESULTS: Post treatment abstinence rates were 71.6% and 53.2% in bupropion SR groups for 7 and 19 weeks of treatment, respectively, (p < 0.001), 63.4% in bupropion SR plus nicotine patch group and 45% (p < 0.001) in nicotine patch group. One year's follow-up abstinence rates were 43.1%, 29.6%, 30.4% and 18.4% (p < 0.05), respectively. Predictors of successful abstinence at the end of therapy included (a) bupropion SR, (b) lower DSM IV symptom score, and (c) lower nicotine addiction, while predictors for sustained abstinence in one year included: (a) bupropion SR, (b) lower nicotine addiction, and (c) smoker's motivation. CONCLUSIONS: Smoking cessation interventions implementing intensive multi component programs and constant smokers' motivation in health care settings of actual practice seem promising for increasing short and long-term abstinence rates. PMID- 19124352 TI - The potential role of natural agents in treatment of airway inflammation. AB - Obstructive airway diseases including asthma, chronic obstructive pulmonary disease and cystic fibrosis present with dyspnea and variety of other symptoms. Physiologically, they are characterized by maximal expiratory flow limitation and pathologically, by inflammation of the airways and the lung parenchyma. Inflammation plays a major role in the gradual worsening of the lung function resulting in worsening symptoms. For many years, scientists focused their efforts in identifying various pathways involved in the chronic inflammation present in these diseases. Further, studies are underway to identify various molecular targets in these pathways for the purpose of developing novel therapeutic agents. Natural agents have been used for thousands of years in various cultures for the treatment of several medical conditions and have mostly proven to be safe. Recent in vivo and in vitro studies show potential anti-inflammatory role for some of the existing natural agents. This review provides an overview of the literature related to the anti-inflammatory effects of some of the natural agents which have potential value in the treatment of inflammatory lung diseases. PMID- 19124353 TI - Worldwide differences on the concept of control of asthma. AB - Asthma is estimated to affect approximately 4-11% of the general population and its prevalence is increasing continuously. A large number of epidemiological studies on the prevalence of asthma and the level of disease control have been carried out, in which the level of control defined by GINA objectives was shown to be significantly unsatisfactory. Frequently, patients accept suboptimal levels of asthma control, mainly because they do not know how asthma is caused and how it can be treated. Apart from having a deep negative impact on patient's daily life, undiagnosed and undercontrolled asthma can also mean an increased economic burden. Therefore, greater attention should be paid to preventive measures, patient education and self-management programmes. Scientific evidence exists that drugs currently available can achieve all or almost all the objectives indicated by international guidelines in the majority of patients, and to improve patients' HRQL by minimizing the impact of the disease and its treatment from the patient's viewpoint. In conclusion, there is a gap between the level of control that could be reached and what is obtained in real life. This must be reduced through a series of interventions in which both doctor and patient are involved. PMID- 19124354 TI - Cough and asthma: the role of inhaled corticosteroids and beta2-agonists. PMID- 19124355 TI - Augmentation therapy for emphysema due to alpha-1-antitrypsin deficiency. AB - Alpha-1 antitrypsin deficiency (AAT) is a hereditary recessive autosomal disease caused by mutations in the AAT gene. This disease is characterized by abnormally low AAT concentrations in plasma, which, in its homozygote form, carries a high risk for the development of early pulmonary emphysema and liver damage. Since the end of the 1980s augmentation therapy with AAT from human plasma has been available for specific treatment of emphysema due to AAT deficiency. Intravenous augmentation therapy has been demonstrated to be safe and weekly infusions of AAT have resulted in plasma AAT concentrations above those considered protective for the lungs. However, life-long weekly infusions are not well accepted by patients, therefore pharmacokinetic studies have been performed to try to individualize the therapeutic regimen in order to obtain adequate trough serum AAT levels with prolonged intervals of administration. Therapeutic regimens administered every two weeks appear to be safe and result in adequate trough serum concentrations, but less-frequent administrations result in trough levels below the target. Alpha 1-antitrypsin deficiency is largely unrecognized and underdiagnosed. The foundation of national and international registries is a valid strategy to increase awareness about the disease and collect information about the natural history of this deficiency. Furthermore, the identification of a large number of patients will allow the development of new clinical trials aimed at finding better treatments for this infrequent condition. PMID- 19124356 TI - FTIR spectroscopic analysis of sputum: preliminary findings on a potential novel diagnostic marker for COPD. AB - COPD is a common, progressively disabling disease and a major health burden worldwide. Fourier transform infrared (FTIR) spectroscopy provides for sensitive analysis of complex biological samples. COPD pathogenesis involves quantitative and qualitative changes in sputum biosynthesis. This first study explores whether FTIR can produce distinct spectral profiles of human sputum, and capture differences between COPD and health. Sputum obtained from 15 COPD patients and 15 healthy volunteers was analysed using FTIR spectroscopy; differences in peak positions, height and configuration were identified and measured. All samples gave reproducible characteristic IR absorption spectra. The most relevant regions identified were the amide and glycogen rich regions, showing crucial spectral differences between health and COPD relating to peak position shifts or intensity alteration. These novel preliminary findings support further exploration of FTIR sputum profiling in a clinical study to determine its potential as a practical method for monitoring COPD. PMID- 19124357 TI - Arformoterol and salmeterol in the treatment of chronic obstructive pulmonary disease: a one year evaluation of safety and tolerance. AB - INTRODUCTION: Concerns have been raised regarding the safety of extended use of long-acting beta2-agonists (LABAs). The safety of arformoterol (50 microg QD), and salmeterol (42 microg BID), was assessed over 12 months in subjects with COPD. The study also examined the occurrence of tolerance with these agents, i.e. whether improvement in airway function diminished or frequency of exacerbations increased with 12-months of use. METHODS: Subjects with COPD (mean FEV1 1.2 L, ~41% predicted) were enrolled in the study and randomized to receive nebulized arformoterol 50 microg QD (n = 528) or salmeterol 42 microg BID (MDI; n = 265) in a prospective, multicenter, open-label, 12-month trial. The frequency of adverse events, COPD exacerbations, and use of short-acting bronchodilator agents were assessed throughout the study period. Pulmonary function was also examined. RESULTS: Among treated subjects, the frequency of adverse events was similar for those taking arformoterol (90.5%) and salmeterol (88.3%). Tremor was more frequent among subjects treated with arformoterol (13.4%) than those treated with salmeterol (1.1%). The frequency of COPD exacerbations did not increase over 12 months for arformoterol and salmeterol (weeks 0-13: 15.7% and 11.7%, respectively; weeks 39-52: 10.0% and 9.4%, respectively). Supplemental ipratropium bromide and rescue racemic albuterol use decreased for both groups by 0.8 to 1.5 actuations/day, decreases that remained stable throughout the 52-week study. Mean predose (trough) FEV1 improved for arformoterol and salmeterol at week 13 (7.1% +/- 17.0 and 7.6% +/- 17.8, respectively) and the improvement continued at week 52 (5.9% and 6.2%, respectively). Mean peak percent predicted postdose FEV1 over the course of the 52-week study declined by about 2% for both treatments, but throughout was higher for arformoterol than for salmeterol. CONCLUSION: In this trial, both arformoterol 50 microg QD and salmeterol 42 microg BID were well tolerated in patients with COPD. Both LABAs produced effective bronchodilation and their use was not associated with the development of clinically meaningful tolerance over a 1-year treatment period. PMID- 19124358 TI - Arterio-venous access in end-stage renal disease patients and pulmonary hypertension. AB - BACKGROUND: The syndrome of pulmonary hypertension (PHT) in end-stage renal disease (ESRD) has been described in patients on chronic hemodialysis (HD) therapy via arterial-venous (A-V) access. However, the exact timing for the development of the PHT is unknown. This study was designed to evaluate changes in pulmonary artery pressure (PAP) following creation of the vascular access. PATIENTS AND METHODS: PAP and cardiac-output (CO) values were recorded in 12 pre dialysis patients without PHT a few months after the access formation, before treatment with HD was started, and the prevalence of PHT was calculated. Clinical data was compared between patients with and without PHT. RESULTS: The systolic PAP values were increased in "ve of the 12 pre-dialysis patients (42%) by 21+/-9 mm Hg to more than 35 mm Hg. Patients with and without PHT differed only in that CO was signi"cantly higher among the former. CONCLUSIONS: The development of PHT following access formation represents a failure of the pulmonary circulation to accommodate the access-mediated elevated CO. Pre-dialysis patients scheduled for access formation should be screened for the presence of sub-clinical PHT. "Positive" patients should proceed to peritoneal dialysis or advance to kidney transplantation; rather than getting access and HD therapy. PMID- 19124359 TI - Exhaled nitric oxide, systemic inflammation, and the spirometric response to inhaled fluticasone propionate in severe chronic obstructive pulmonary disease: a prospective study. AB - BACKGROUND: A subset of patients with chronic obstructive pulmonary disease (COPD) may respond more favorably to inhaled corticosteroids (ICS), but no simple method is currently utilized to predict the presence or absence of ICS responses in patients with COPD.We evaluated the ability of exhaled nitric oxide (FENO) and serum inflammatory markers (C-reactive protein [CRP], interleukin-6 [IL-6], and interleukin-8 [IL-8]) to independently predict spirometric responses to ICS in patients with COPD. METHODS: Among 60 ex-smokers with severe COPD (mean FEV1 1.07 L, 36% of predicted), we conducted a single-arm, open-label study. Participants spent four weeks free of any ICS, followed by four weeks of ICS use (fluticasone propionate 500 mcg twice daily). FENO, CRP, IL-6, IL-8, and pre-bronchodilator spirometry were measured immediately before and after the four weeks of ICS use. RESULTS: Baseline FENO, CRP, IL-6, and IL-8 showed no correlations to FEV1 responses to ICS. ICS responders (increase in FEV1 > or = 200 mL after four weeks of ICS) did have significantly higher baseline FENO levels compared with non responders (46.5 parts per billion [ppb] vs. 25 ppb, p = 0.028). The receiver operating characteristic curve for FENO to discriminate responders from non responders had an area under curve of 0.72. Baseline serum inflammatory markers did not differ between responders and non-responders. CONCLUSION: In ex-smokers with severe COPD, a measure of local pulmonary inflammation, FENO, may be more closely associated with FEV1 responses to four weeks of ICS than are standard markers of systemic inflammation, serum CRP, IL-6, and IL-8. PMID- 19124360 TI - Surfactant protein D: a lung specific biomarker in COPD? AB - A major impediment in the development of novel drugs for chronic obstructive pulmonary disease (COPD) has been the scarcity of a well-validated, robust, and easily obtainable intermediate end point such as serum biomarkers. To date the best serum biomarkers in COPD have been non-speci"c pro-in"ammatory molecules synthesized largely by extra-pulmonary organs. In COPD, an ideal biomarker would be one that (1) was produced mostly in the lungs (and was reliably measurable in the peripheral circulation using commercially available kits), (2) changed with the clinical status of patients or with relevant exposures; and (3) had inherent functional attributes that suggested a possible causal role in the pathogenesis of the disease. In this paper, we review one promising systemic biomarker that ful"lls some of these criteria, surfactant protein D (SPD). PMID- 19124361 TI - New perspectives in the treatment of idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is the most frequent idiopathic interstitial pneumonia with a prevalence ranging from 5 to 15 per 100,000 persons, and above 175 per 100,000 in the older population. IPF is a relentlessly progressive fibrotic lung disorder leading to death within a median duration of 3 years. It was hypothesized in the 1970s that pulmonary fibrosis initiates as an "alveolitis" progressing to interstitial fibrosis with connective tissue deposition, derangement of the lung architecture and functional impairment. However, in vitro studies indicated that alveolar/bronchiolar injured epithelial cells can drive the fibrotic process in the absence of macrophages and with minimal inflammation. This, together with the inability of classic immunosuppressive therapy to cure IPF, generated new pathogenesis paradigms and intense research into the role of the lack or the excessive production of anti fibrotic or profibrotic mediators, oxidant injury, exaggerated coagulation, thus leading to investigate new treatment strategies. Preliminary results of some of such trials have shown significant reductions in lung function decline, disease exacerbation and mortality. PMID- 19124362 TI - Treating patients with respiratory disease who smoke. AB - The high prevalence of cigarette smoking in patients with respiratory disease puts them at risk of developing clinically important drug interactions. Cigarette smoking reduces the therapeutic response to certain drugs such as theophyllines through the induction of hepatic cytochrome P450 isoenzymes. Smokers with asthma and patients with COPD have reduced sensitivity to corticosteroids, possibly due to non-eosinophilic airway inflammation, altered glucocorticoid receptor activity or reduced histone deacetylase activity. Although all smokers should be encouraged to stop smoking, there is limited information on the influence of smoking cessation on the therapeutic and anti-inflammatory effects of a number of the drugs used in the treatment of respiratory disease. PMID- 19124363 TI - Unifying the genetics, co-morbidities and management of COPD. PMID- 19124365 TI - Airway smooth muscle as a model for new investigative drugs in asthma. AB - Bronchial asthma as such exists because airway smooth muscle (ASM) contracts excessively in response to various stimuli. After several decades during which research was mainly focused on airway inflammation, increasing attention is now being paid to a possible abnormal behaviour of ASM. Thus, ASM is regarded as a major target for anti-asthma treatments. This review first describes the mechanisms of ASM contraction and airway hyperresponsiveness, through cellular, animal and human models. The developments of new drugs targeting extra and/or intracellular pathway of ASM contraction are discussed. PMID- 19124364 TI - Role of phosphodiesterases in modulation of BKCa channels in hypertensive pulmonary arterial smooth muscle. AB - BKCa channels regulate pulmonary arterial pressure, and protein kinase C (PKC) inhibits BK(Ca) channels, but little is known about PKC-mediated modulation of BKCa channel activity in pulmonary arterial smooth muscle. Studies were carried out to determine mechanisms of PKC modulation of BKCa channel activity in pulmonary arterial smooth muscle cells (PASMC) of the fawn-hooded rat (FHR), an animal model of pulmonary hypertension. Forskolin opened BKCa channels in FHR PASMC, which was blocked by PKC activation, and reversed by the phosphodiesterase (PDE) inhibitors IBMX, milrinone, and zaprinast. PDE inhibition also blocked the vasoconstrictor response to PKC activation in FHR pulmonary arteries. These results indicate that PKC inhibits cAMP-induced activation of BKCa channels and causes pulmonary vasoconstriction in hypertensive pulmonary arterial smooth muscle via PDE, which further suggests PDE inhibitors for treatment of pulmonary hypertension. PMID- 19124366 TI - Control of asthma. AB - Control is the aim of asthma management and clinical trials show that control can be achieved in the majority of patients. However, population surveys show that poorly controlled asthma still imposes a considerable burden. This fact has led to a re-evaluation of the international asthma guidelines and the updated 2006 and 2007 GINA and NAEPP guidelines suggest that the level of control should be used as the key feature for the classification and management of asthma. Furthermore, in the latest guidelines, a clearer definition of control is given and new tools for the assessment and monitoring of control are instituted. In order to achieve asthma control, not only relevant pharmacological treatment but, the establishment of a good patient-doctor relationship, proper education of the asthmatic patient, reduction of exposure to triggers and treatment of co morbidities are pivotal issues and must be ensured. PMID- 19124367 TI - Evaluating combination therapies for asthma: pros, cons, and comparative benefits. AB - Combination therapies with inhaled corticosteroids (ICS) and either long-acting beta2 agonists (LABA) or leukotriene receptor antagonists (LTRA) are commonly used to help patients maintain control of their asthma. LABA and LTRA have different mechanisms of action and both provide complementary benefits when combined with ICS. This paper compares the two regimens based on recent clinical trial data, evaluates their efficacy on various clinical and quality of life outcomes, and discusses the importance of therapy choice in people with specific asthma phenotypes. The potential of new dosing strategies, including adjustable maintenance therapy and single-inhaler maintenance and reliever therapy is also reviewed. Given the variety of outcomes, phenotypes, and treatment strategies that must be considered, the importance of individualized management is emphasized. PMID- 19124368 TI - Tissue remodeling and angiogenesis in asthma: the role of the eosinophil. AB - This review covers the role of eosinophils in asthma-associated tissue remodeling and angiogenesis focusing on angiogenesis which is a recently discovered feature of asthma. In addition, novel directions for eosinophil-targeted/angiogenesis targeted pharmacological intervention are discussed as new approaches in the treatment of asthma. PMID- 19124369 TI - Stem cell therapy: the great promise in lung disease. AB - Lung injuries are leading causes of morbidity and mortality worldwide. Pulmonary diseases such as asthma or chronic obstructive pulmonary disease characterized by loss of lung elasticity, small airway tethers, and luminal obstruction with inflammatory mucoid secretions, or idiopathic pulmonary fibrosis characterized by excessive matrix deposition and destruction of the normal lung architecture, have essentially symptomatic treatments and their management is costly to the health care system.Regeneration of tissue by stem cells from endogenous, exogenous, and even genetically modified cells is a promising novel therapy. The use of adult stem cells to help with lung regeneration and repair could be a newer technology in clinical and regenerative medicine. In fact, different studies have shown that bone marrow progenitor cells contribute to repair and remodeling of lung in animal models of progressive pulmonary hypertension.Therefore, lung stem cell biology may provide novel approaches to therapy and could represent a great promise for the future of molecular medicine. In fact, several diseases can be slowed or even blocked by stem cell transplantation. PMID- 19124370 TI - Therapeutic perspectives in bronchial vascular remodeling in COPD. AB - COPD may be characterized by significant changes in airway mucosal blood vessels, which may contribute to bronchial airway remodeling. The airway wall is more vascularized in COPD patients than in healthy subjects, though this phenomenon is less evident than in asthmatic patients. The vascular mucosal changes in the airways of patients with COPD are strictly linked to the inflammatory processes. The cellular mechanisms responsible for the microvascular changes are still unclear, however, pro-angiogenic factors, such as VEGF, TGF-beta, FGF, and proteolytic enzymes such as MMPs, may play a role. Up to now, the clinical and functional consequences of this phenomenon and the therapeutic approach have been scarcely investigated. Inhaled corticosteroids seem to have positive effects, by reducing the vascular area and growth factor expression. Specific antagonists to VEGF, TGF- beta, FGF, and MMPs could beneficially control chronic airway inflammation and vascular remodeling in COPD as well as slow down the progression of the disease. Several of these growth factor antagonists are being evaluated and some seem to be effective in reducing vascularity, however further studies are required to ascertain whether or not these antagonists may play a role in COPD therapy. PMID- 19124371 TI - Adverse events associated with intranasal influenza vaccine in the United States. AB - METHODS: The goal of this review was to analyze the post-vaccination adverse effects associated with Live Attenuated Influenza Vaccine (LAIV) compared to those of the Injectable Trivalent Inactivated Vaccine (TIV) in patients with egg allergy and asthma. PubMed, Ovid, and Google searches were conducted. Searched articles relating to allergic reactions, asthma or food allergy, and LAIV were reviewed. RESULTS: Similar or superior efficacy of LAIV compared to TIV was reported by various studies. Respiratory symptoms were the most common adverse events following both vaccinations. Although no evidence was found of a direct causal relationship between intranasal influenza vaccine and anaphylaxis due to egg allergy, a number of cases of anaphylaxis were reported. To date, there are no studies directly comparing the frequency of anaphylactic reactions between the two vaccines. CONCLUSIONS: The safety of LAIV in individuals with unstable asthma and egg allergy has not been established and it should be avoided in these populations. For patients with unstable asthma, TIV should remain the therapy of choice. PMID- 19124372 TI - Long-term safety of nebulized formoterol: results of a twelve-month open-label clinical trial. AB - Formoterol fumarate is a long-acting beta2-agonist that is an effective bronchodilator for the maintenance management of patients with chronic obstructive pulmonary disease. The safety profile of the newly developed nebulized formoterol was evaluated over a twelve-month period in an open-label, active-control study. After completing a twelve-week double-blind double-dummy period, 569 subjects with chronic obstructive pulmonary disease entered an open label extension study and received twice-daily 20 microg formoterol fumarate inhalation solution for nebulization (FFIS) or 12 microg formoterol fumarate dry powder inhalation (FA) for 52 weeks. Most of the FFIS-treated subjects (86%) completed at least six months of open-label treatment with over 90% compliance, comparable to the FA group (88%). RESULTS: of safety monitoring for adverse events, laboratory values, and cardiac changes were similar between treatment groups. Three hundred forty (73%) of FFIS-treated subjects and 83 (78%) of FA treated subjects experienced an adverse event over the course of the study, the majority of which were mild to moderate and considered unrelated to treatment. COPD exacerbation occurred in 15.8% of FFIS-treated and 17.9% of FA-treated subjects. Deaths, serious adverse events, and discontinuations for adverse events occurred in 1.3, 16.2, and 5.4% of the nebulized group versus 1.9, 17.9, and 7.5% of the inhaled group, respectively. There were no clinically important changes from baseline in laboratory tests, including serum potassium and glucose, or vital signs and no treatment-related increases in cardiac arrhythmias, heart rate, or QTc prolongation. We conclude that nebulized formoterol fumarate twice daily is well tolerated over long-term treatment in moderate-to-severe COPD subjects and has a similar safety profile to the DPI formulation. PMID- 19124373 TI - Prulifloxacin in the treatment of acute exacerbations of COPD in cigarette smokers. AB - Smoking is associated with an increased risk of respiratory tract infection in adults likely because components in the smoke might alter properties of the epithelial cell surface. In studies with smokers suffering from acute exacerbations of COPD (AECOPD), the most common bacterial pathogens found were mainly Haemophilus influenzae, but also Streptococcus pneumoniae, Staphylococcus aureus, and Moraxella catarrhalis. Therefore, antibiotics should be effective against such possible pathogens. Prulifloxacin has demonstrated in vitro activity against all these pathogens. We designed the present study to evaluate the efficacy of prulifloxacin in the treatment of AECOPD in cigarette smokers. We enrolled 61 consecutive smokers hospitalized or out-patients of either sex with symptoms and signs compatible with the usual diagnosis criteria for AECOPD. Haemophilus influenzae was the most common bacterial species isolated in the sputum (in 42.6% of the total sample), followed by S. pneumoniae (16.5%), S. aureus (14.7%), M. catarrhalis (11.5%), and others (14.7%). Prulifloxacin 600 mg was given orally once daily for 10 days. Clinical success was observed in 91.8% of patients (67.2% cured and 24.6% improved). Bacteriological eradication rate of H. influenzae was 100%. Persistent pathogens were S. pneumoniae (2 out of 10), S. aureus (1 out of 8), M. catarrhalis (1 out of 7), and P. aeruginosa (1 out of 3). This study seems to indicate that prulifloxacin is of particular value in the treatment of AECOPD in cigarette smokers. PMID- 19124374 TI - Oxidant--antioxidant imbalance in asthma: scientific evidence, epidemiological data and possible therapeutic options. AB - Prevalence of asthma has increased considerably in recent decades throughout the world especially in developed countries. Airway inflammation is thought to be prime cause for repeated episodes of airway obstruction in asthmatics. Several studies have shown that reactive oxygen species (ROS) play a key role in initiation as well as amplification of inflammation in asthmatic airways. Excessive ROS production in asthma leads to alteration in key enzymatic as well as nonenzymatic antioxidants such as glutathione, vitamins C and E, beta carotene, uric acid, thioredoxin, superoxide dismutases, catalase, and glutathione peroxidases leading to oxidant-antioxidant imbalance in airways. Oxidant-antioxidant imbalance leads to pathophysiological effects associated with asthma such as vascular permeability, mucus hypersecretion, smooth muscle contraction, and epithelial shedding. Epidemiological data also support the scientific evidence of oxidant-antioxidant imbalance in asthmatics. Therefore, the supplementation of antioxidants to boost the endogenous antioxidants or scavenge excessive ROS production could be utilized to dampen/prevent the inflammatory response in asthma by restoring oxidant-antioxidant balance. This review summarizes the scientific and epidemiological evidence linking asthma with oxidant-antioxidant imbalance and possible antioxidant strategies that can be used therapeutically for better management of asthma. PMID- 19124375 TI - Update on lung transplantation. AB - Lung transplantation is a suitable treatment to improve the quality of life and the prognosis of patients with various end-stage pulmonary diseases. There is a shortage of organs and the number of patients on waiting list is exceeding the number of transplants per year approximately two-fold in most countries. Strategies to increase the donor pool are therefore crucial. The long-term prognosis after lung transplantation is not yet as good as for other transplantation procedures and special complications should be taken into account. Aside from infections chronic organ dysfunction is the main cause of death and the most important prognosis limiting factor. Other typical problems are rejection, bronchus complications, and primary transplant dysfunction. However, the results of large centers show promising improvements in recent years. PMID- 19124376 TI - Therapeutic advances in pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) is characterized by vasoconstriction, in situ thrombosis, and vascular remodeling of small pulmonary arteries inducing increased pulmonary arterial resistance. Conventional treatment is based on life style modification and nonspecific treatment (warfarine, diuretics, oxygen). Calcium channel blockers are vasodilatators that have been shown to be of great efficacy in a very specific subpopulation of patients with PAH. For the majority of patients, specific PAH therapies are still lacking. Numerous studies evaluating prostacyclin agonists, endothelin-receptor antagonists, and phosphodiesterase type 5 inhibitors are now available to guide therapeutic choices. Despite those important advances there is still no cure for PAH. Fortunately, research is ongoing and many drugs show promises. PMID- 19124377 TI - Erdosteine enhances airway response to salbutamol in patients with mild-to moderate COPD. AB - BACKGROUND: Oxidative stress is presumed to impair beta-adenoceptor function and airway patency. Erdosteine (E), a mucomodulatory compound, has shown important antioxidant properties. METHODS: The objective was to assess the effect of antioxidant interventions on short-term airway response to salbutamol in non reversible mild-to-moderate COPD patients. Thirty COPD patients (GOLD class 1-2), current smoker (>or=10 pack/year), randomly received E 300 mg, N-acetylcysteine (NAC) 600 mg, or placebo, twice daily for ten days. Reversibility to salbutamol 200 microg was tested in baseline, after four and ten days of each treatment. ROS and 8-isoprostane blood levels were measured on the same days. Between-treatment comparison was performed by ANOVA and t-test or Wilcoxon test, and p<0.05 assumed. E enhanced FEV1 reversibility after four and ten days significantly (+5.1% and +5.0%; both p<0.01 vs. placebo), while NAC only showed a transient effect at day 4 (+3.0%, p<0.05), but not at day 10 (+1.3%, p = ns). RESULTS: E and NAC caused significant drops in ROS blood levels after four and ten days (p<0.001 and p<0.0001 vs. placebo). In contrast to NAC, E lowered 8-isoprostane levels substantially for ten days (p = 0.017 and p = 0.0004 vs. placebo, respectively). Only E restored significantly short-term reversibility in COPD patients previously unresponsive to beta(2)-adrenergics. CONCLUSIONS: This effect seems more related to the peculiar protection against lipid peroxidation rather than to the scavenging activity, which proves equal to that of NAC. E provides a sort of indirect bronchodilation through 're-sensitisation' of beta( 2) adrenoceptors. Once confirmed in further controlled studies, it may be useful in long-term treatment of COPD. PMID- 19124378 TI - Differences in the pharmacodynamics of budesonide/formoterol and salmeterol/fluticasone reflect differences in their therapeutic usefulness in asthma. AB - Although the available inhaled corticosteroid (ICS)/long-acting beta( 2)-agonist (LABA) combinations principally work in a similar fashion, they differ in several important ways, leading to different efficacy. The ICS/LABA combination product budesonide/formoterol can be used as both maintenance and reliever therapy, providing a fixed maintenance dose, which does not change, and replacing short acting beta(2)-agonists as relievers thereby allowing intervention to address the underlying inflammation at the earliest sign of symptomatic worsening. This approach is not suitable for other combination products such as salmeterol/fluticasone. Here we review the pharmacological differences of budesonide/ formoterol and salmeterol/fluticasone that permit the use of budesonide/formoterol as both maintenance and reliever therapy. PMID- 19124379 TI - Pharmacotherapy for smoking cessation. AB - Tobacco dependence is a chronic relapsing disease that needs continuous treatment. In combination with behavioural support, pharmacotherapy is a proven key component for supporting smoking cessation. Effective drugs are available and recommended: nicotine replacement therapy (NRT), bupropion and varenicline. Much research into new pharmacological approaches is ongoing, combining 'old' and 'new' drugs and personalizing a pharmacological treatment for a single smoker/patient; other new medications and vaccines are in development. Overall, pharmacotherapy seems to have efficacy and cost-effectiveness in real life, thus physicians should become familiar with these medicines. Further efforts should be aimed at optimizing treatment management and increasing smoking cessation rates in the general population. PMID- 19124380 TI - Therapies for interstitial lung disease: past, present and future. AB - As our understanding of the pathobiology and natural history of the various forms of interstitial lung disease (ILD) has evolved, so have our approaches to treating this heterogeneous group of lung disorders. The earliest pharmacologic agents used to treat various forms of ILD were corticosteroids, and corticosteroids are currently the mainstay of therapy for many forms of ILD. However, it has become clear that corticosteroids and other anti-inflammatory agents lack efficacy for many forms of ILD, such as idiopathic pulmonary fibrosis (IPF), and newer therapies that are in clinical trials target the fibrogenic process and/or secondary pulmonary hypertension (PH) that is present in various forms of fibrotic lung disease. Novel therapies, such as the use of biologic agents (antibodies and cell cycle inhibitors) or stem cell therapies will undoubtedly evolve as new research is performed and clinical trials are undertaken. Lung transplantation remains an option for advanced lung disease that is progressive and unresponsive to non-surgical therapies. PMID- 19124381 TI - Surfactant respiratory therapy using Surfaxin/sinapultide. AB - Respiratory distress syndrome (RDS) is a leading cause of mortality and morbidity in preterm infants. Surfactant replacement therapy has been widely used to prevent and treat RDS in these newborns and has now become a standard of care. First-generation synthetic surfactants such as Exosurf did not contain any surfactant protein. This disadvantage was overcome with animal-derived surfactant preparations which contain specific proteins but has the limitation of being derived from animal sources. This has led to development of newer synthetic surfactants such as lucinactant (Surfaxin, Discovery Laboratories, Philadelphia) which contains the protein B mimic synthetic peptide, sinapultide. Recent phase 3 clinical trials with Surfaxin show promising results with similar efficacy as animal derived surfactants and yet avoiding the disadvantage associated with animal products. The purpose of this paper is to summarise results of recent clinical trials of Surfaxin use in newborns with RDS. PMID- 19124383 TI - Pharmacotherapy of obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is associated with serious comorbid illnesses and diminished quality of life. At this time, continuous positive airway pressure (CPAP) therapy is the treatment of choice. However, only half of those individuals who accept CPAP are still using it at the end of one year. Furthermore, efficacy for improving self-reported sleepiness appears to be greater for patients with severe sleep apnea and severe sleepiness than other patient groups. Some patients, notwithstanding optimized therapy and therapeutic adherence continue experiencing excessive daytime somnolence. Consequently, other treatment modalities have developed, including oral appliances, surgery and pharmacotherapy. It is widely believed, albeit not empirically demonstrated, that an effective medication to treat OSA would elicit better acceptance and adherence than having to use a machine for many hours on a nightly basis. Nonetheless, paucity of data (i.e. lack of large-scale randomized controlled trials), variability of perceived and actual benefits, and adverse side-effects of the drugs thus far tested have prevented the use of pharmacotherapy until now. In this paper we review the outcome data from published trials designed to evaluate efficacy and safety of various medications proposed for treating obstructive sleep apnea. PMID- 19124384 TI - Novel targets in the management of pneumonia. AB - Community-acquired pneumonia (CAP) is the leading cause of death from infectious diseases in the US. It accounts each year for 500,000 hospitalizations and 45,000 deaths and represents one of the most common causes of intensive care unit (ICU) admission. The mortality rate due to severe CAP has shown little improvement in the past three decades, remaining between 21% and 58% in patients admitted to the intensive care unit. Antimicrobial agents are the cornerstone of therapy against CAP, but there are some novel antibiotic and nonantibiotic therapies that have been recently tested that may potentially impact outcomes of patients with severe CAP. We will review the most recent data regarding novel therapies in patients with the highest risk of death such as those with severe CAP. PMID- 19124385 TI - Managing TB in the 21st century: existing and novel drug therapies. AB - With an estimated one-third of the world's population infected with Mycobacterium tuberculosis (MTB) and approximately 1.6 million deaths in 2006 attributed to tuberculosis (TB) world-wide, TB remains a major public health concern today. Considerable advances have been made in the effective treatment of TB, in particular with the adoption of directly observed therapy short course (DOTS), in national TB control programs, but in spite of this the currently available regimens are suboptimal. The long courses of therapy required, together with significant medication side-effects and resulting difficulties with adherence to therapy all contribute to increasing problems with emerging drug resistance. There is thus an urgent need for new antituberculous drug development, especially to enable effective shorter course therapy for drug-susceptible and resistant TB, to find effective drugs for treatment of drug-resistant TB, to shorten therapy for latent TB infection and to reduce drug interactions in combination with antiretroviral therapy, a major issue in the treatment of HIV co-infected patients. This review will attempt to summarize the current recommendations for treatment of TB and then describe the most promising new antimicrobials with activity against MTB, focusing on the ones currently undergoing clinical trials. PMID- 19124386 TI - Omalizumab in the treatment of severe asthma: efficacy and current problems. AB - Omalizumab is a humanized monoclonal anti-IgE antibody recently approved for the treatment of severe allergic asthma. This drug inhibits allergic responses by binding to serum IgE, thus preventing their interactions with cellular IgE receptors. Omalizumab is also capable of downregulating the expression of high affinity IgE receptors on inflammatory cells, as well as the numbers of eosinophils in both blood and induced sputum. The clinical effects of omalizumab include relevant improvements in respiratory symptoms and quality of life, paralleled by a marked reduction of asthma exacerbations, emergency room visits, and use of systemic corticosteroids and rescue bronchodilators. Omalizumab is relatively well tolerated, and only rarely induces anaphylactic reactions. Therefore, this drug represents a valid option as add-on therapy for patients with severe persistent allergic asthma, inadequately controlled by high doses of standard inhaled treatments. PMID- 19124382 TI - Antioxidant therapeutic advances in COPD. AB - Chronic obstructive pulmonary disease (COPD) is associated with a high incidence of morbidity and mortality. Cigarette smoke-induced oxidative stress is intimately associated with the progression and exacerbation of COPD and therefore targeting oxidative stress with antioxidants or boosting the endogenous levels of antioxidants is likely to have beneficial outcome in the treatment of COPD. Among the various antioxidants tried so far, thiol antioxidants and mucolytic agents, such as glutathione, N-acetyl-L-cysteine, N-acystelyn, erdosteine, fudosteine and carbocysteine; Nrf2 activators; and dietary polyphenols (curcumin, resveratrol, and green tea catechins/quercetin) have been reported to increase intracellular thiol status along with induction of GSH biosynthesis. Such an elevation in the thiol status in turn leads to detoxification of free radicals and oxidants as well as inhibition of ongoing inflammatory responses. In addition, specific spin traps, such as alpha-phenyl-N-tert-butyl nitrone, a catalytic antioxidant (ECSOD mimetic), porphyrins (AEOL 10150 and AEOL 10113), and a SOD mimetic M40419 have also been reported to inhibit cigarette smoke-induced inflammatory responses in vivo in the lung. Since a variety of oxidants, free radicals and aldehydes are implicated in the pathogenesis of COPD, it is possible that therapeutic administration of multiple antioxidants and mucolytics will be effective in management of COPD. However, a successful outcome will critically depend upon the choice of antioxidant therapy for a particular clinical phenotype of COPD, whose pathophysiology should be first properly understood. This article will review the various approaches adopted to enhance lung antioxidant levels, antioxidant therapeutic advances and recent past clinical trials of antioxidant compounds in COPD. PMID- 19124387 TI - Pre-eclampsia: the pivotal role of the placenta in its pathophysiology and markers for early detection. AB - Pre-eclampsia is the second leading cause of maternal morbidity and mortality in the United States. Infants born to affected mothers face a five-fold increase in death rate [Lain and Roberts 2002; National Heart Lung and Blood Institute 2001]. Although pre-eclampsia has been recognized by physicians for millennia, relatively little is known about its pathogenesis or prevention. Predicting its development is often extremely difficult, perhaps leading the Greeks to use the name 'eklampsis' meaning lightening. Recent studies provide novel insights into the role of the placenta in the development of pre-eclampsia and demonstrate novel markers to assist in predicting the onset of disease and potential therapeutic targets. Following an introduction which highlights the classification of hypertensive disorders of pregnancy and defines incidence and adverse outcomes of pre-eclampsia, this manuscript will discuss the role of the placenta in the pathophysiology of pre-eclampsia and recent markers that may predict its onset. PMID- 19124388 TI - Drug-eluting stent thrombosis. AB - Stent thrombosis is a rare complication following stent implantation; if it occurs, however, it is associated with a high morbidity and mortality. Despite reduced rates of restenosis, drug-eluting stents (DES) have not reduced the incidence of stent thrombosis as compared with bare-metal stents (BMS). Patient-, lesion-, and procedure-related factors as well as thrombogenicity of the stent itself are involved in the pathogenesis of stent thrombosis. Furthermore, early cessation of dual antiplatelet therapy correlates with an increased risk of stent thrombosis. This review focuses on clinical evidence and pathophysiological mechanisms of stent thrombosis with DES, particularly highlighting prothrombotic effects of the stent itself. PMID- 19124389 TI - Electrophysiological changes of cardiac function during antidepressant treatment. AB - Some antidepressant agents can cause electrophysiological changes of cardiac function leading to ventricular arrhythmias and sudden death. However, antidepressants have also protective effects on the heart through their capacity to modulate cardiac autonomic-mediated physiological responses. Heart rate variability and QTc length are two strictly linked parameters that allow us to appreciate the effects of different drugs on cardiac physiology. Heart rate variability reflects functioning of the autonomic nervous system and possibly also regulation by the limbic system. Autonomic regulation of cardiac activity influences also cardiac repolarization and QT length, both directly and via its effects on heart rate. In this review we present the methodologies adopted to study the effect of antidepressant drugs on QT length and heart rate variability and we summarize data on electrophysiological changes related to antidepressant treatment. Clinical implications for the choice of different antidepressants in different clinical populations are discussed. PMID- 19124390 TI - Effects of alcohol on atrial fibrillation: myths and truths. AB - Alcohol is the most consumed drug worldwide. Both acute and chronic alcohol use have been associated with cardiac arrhythmias, in particular atrial fibrillation, or so-called 'holiday heart syndrome'. Epidemiological, clinical and experimental studies have attempted to elucidate the mechanisms involved in this association. However, because most of these studies have shown conflicting results, the connection between ethanol and atrial arrhythmias remains controversial. Historical, epidemiological and pharmacological aspects of alcohol, as well as recent concepts on atrial fibrillation are reviewed. We then examine the literature and provide a critical point of view on the still elusive association between alcohol and atrial fibrillation. PMID- 19124392 TI - The metabolic syndrome as an endocrine disease: is there an effective pharmacotherapeutic strategy optimally targeting the pathogenesis? AB - The metabolic syndrome (MetS) represents a combination of cardiovascular risk determinants such as obesity, insulin resistance and lipid abnormalities such as hypertriglyceridemia, increased free fatty acids, low high-density-cholesterol and hypertension. As a multiple component condition it imparts a doubling of relative risk for atherosclerotic cardiovascular disease (ASCVD). It is currently controversial which component of the syndrome carries what weight. There is even a considerable debate whether the risk for ASCVD is greater in patients diagnosed with MetS than that by the individual risk factors. At present, no unifying pathogenetic mechanism can explain the metabolic syndrome and there is no unique treatment for it. This review summarizes and critically reviews the currently available clinical and scientific evidence for the concept that the MetS is causally an endocrine disease and discusses pharmacotherapeutic strategies targeting the pathogenesis rather than single symptoms of the cluster. PMID- 19124393 TI - Renin-angiotensin-aldosterone system intervention in the cardiometabolic syndrome and cardio-renal protection. AB - The metabolic syndrome, also known as the cardiometabolic syndrome (CMS), is a state of metabolic and vascular dysregulation that is associated with activation of the renin-angiotensin-aldosterone system (RAAS). Clinical components of the CMS include central or visceral obesity, hypertension (HTN), dyslipidemia, insulin resistance/hyperinsulinemia, and microalbuminuria that collectively convey increases in oxidative stress, inflammation, and subsequent endothelial dysfunction. The cardio-renal inflammation and oxidative stress enhanced in the CMS increases the risk for cardiovascular disease (CVD) and renal disease end points such as stroke, congestive heart failure, and chronic kidney disease (CKD). The development of proteinuria is known to herald progressive kidney disease (e.g. CKD) and both are now well accepted as CVD risk factors. Evidence suggests a role for visceral obesity, insulin resistance/hyperinsulinemia, HTN, and other components of the CMS lead to an increased risk for proteinuria and progressive loss of renal function. Intervention with agents that block the RAAS (e.g. ACE inhibitors and Angiotensin type 1 receptor blockers) have been shown to reduce proteinuria, CKD progression, and CVD events. Herein, we will examine the relationship between RAAS intervention and reductions in CKD and CVD events. PMID- 19124394 TI - Genetics of the cardiometabolic syndrome: new insights and therapeutic implications. AB - Although the definition of the phenotype is imprecise, cardiometabolic syndrome (CMS) includes a constellation of complex diseases such as type 2 diabetes, dislipidemias, central obesity and hypertension, proinflammatory and prothrombotic states, ovarian polycystosis and fatty liver. The genetics of each disease is complex in itself and varies in spectrum from monogenic and syndromic models of inheritance, usually rare, to the most common polygenic and multifactorial forms. In addition, human studies using the candidate-gene approach indicate that common genetic variants of several genes are associated with the development of CMS. Genome-wide scans have also provided several chromosomal regions associated with some of the components of CMS. In addition, through comparative genomics animal models can generate a map for candidate loci in humans and a promising approach is offered by bioinformatic tools for gene prioritization. Lastly, the involvement of genes whose products are already the targets for approved drugs, such as SLC6A4, PPARalpha and PPARgamma , in the development of CMS suggests new avenues for CMS pharmacological treatment. PMID- 19124395 TI - HMG CoA reductase inhibitors and renoprotection: the weight of the evidence. AB - Dyslipidemia and the contributions of oxidized low-density lipoproteins (ox-LDL) are independent cardiovascular risk factors. There is growing evidence that dyslipidemia contributes not only to cardiovascular disease but also to the progressive decline of renal function in diabetic and non-diabetic kidney disease. Ox-LDL, by generating inflammation and oxidative stress, contributes to a pro-atherogenic mileu and leads to endothelial dysfunction, subsequent glomerular filtration barrier damage, and progressive renal injury. Chronic kidney disease (CKD), in turn, induces deleterious effects on lipid metabolism. Therefore, by inhibiting cholesterol synthesis and reducing ox-LDL, HMG CoA reductase inhibitors (statins) are attractive therapeutic options to preserve renal function. Current evidence demonstrates a reduction in cardiovascular risk and improved renal outcomes especially in patients with mild to moderate impairment of renal function. Evidence supports a beneficial role of statins thought to extend beyond their lipid-lowering effect, referred to as pleiotropic actions. These actions include modulatory effects on inflammation, oxidative stress and thrombosis, derived from their ability to prevent the formation of isoprenoid intermediates involved in cellular signaling, posttranslational modification of proteins and cellular function. This translates to potential reductions in the rate of decline in GFR in CKD and adverse effects of type 2 diabetes mellitus in the kidney. This review examines the role of statins for reno-protection as well as cardiovascular benefit in patients with CKD. PMID- 19124396 TI - Obesity and cardiometabolic syndrome in children. AB - The cardiometabolic syndrome is highly prevalent among overweight youth. The risk of developing the cardiometabolic syndrome is likely triggered or exacerbated by concurrent obesity, unhealthy lifestyle/eating habits, and hormonal changes (puberty). Current screening recommendations include measurement of blood pressure, fasting insulin and glucose, and total cholesterol. However, limiting assessments to these measures underestimates cardiometabolic risk in overweight youth, particularly minorities. Early identification of cardiometabolic risk in its incipient stages may justify early and more aggressive intervention to prevent progression and complications. This review provides rationale for additional assessments to determine cardiometabolic risk in overweight youth and recommends treatment options. PMID- 19124397 TI - Hypertension and the cardiometabolic syndrome in Chile: a review of concepts and consequences for the developing world. AB - We review some recent developments regarding the concept of cardiometabolic syndrome and its relation with hypertension and overall cardiovascular disease risk. We emphasize how this new clinical entity has helped to understand multimorbidity in chronic diseases. This concept has important consequences for individual patient treatment as well as public health policy. The challenge derived from cardiovascular disease and other chronic conditions is increasing worldwide, but the highest burden is located in the developing world. Thus, new and cost-effective approaches are needed for diseases that are mainly occurring in the poorest and less educated populations. We illustrate this situation analyzing hypertension and cardiometabolic syndrome data derived from a recent national health survey in Chile. PMID- 19124399 TI - Therapy for the altered mineral metabolism of chronic kidney disease: implications for vascular calcification. AB - The development of chronic kidney disease (CKD) is associated with poor outcomes largely due an increased likelihood of adverse cardiovascular events. Many factors are playing a role in cardiovascular disease in CKD including the development of vascular calcification (VC). Studies have indeed shown that the presence of VC is associated with decreased survival among patients with CKD. The pathogenesis of VC is itself multi-factorial. LikeLy playing a significant role is the altered mineral metabolism of CKD. Management of the altered mineral metabolism in CKD is quite difficult and may also play a role in the pathogenesis of VC. In this manuscript, we will review the pathogenesis of the altered mineral metabolism of CKD, its management, and how both may play a role in the development of VC. PMID- 19124400 TI - The role of noninvasive hemodynamic monitoring in the evaluation and treatment of hypertension. AB - Advances in the understanding of the mechanisms accounting for the elevation of arterial pressure in essential hypertension suggest that there is value in assessing the relative contribution of hemodynamic factors in tailoring specific therapies to control arterial pressure. The non-invasive method of impedance cardiography (ICG) to measure hemodynamic abnormalities in hypertensive patients has emerged as a valuable adjuvant in the decision-making process of selecting antihypertensive agents. The technique is both accurate and reproducible in delineating the hemodynamic mechanisms of hypertension, comparing age-and gender related changes in hemodynamics, detecting the presence of left ventricular dysfunction, and demonstrating clinically significant improvement in blood pressure control using ICG-guided therapy. PMID- 19124398 TI - Carotid intima-media thickness and plaque volume changes following 2-year angiotensin II-receptor blockade. The Multicentre Olmesartan atherosclerosis Regression Evaluation (MORE) study. AB - OBJECTIVE: The Multicentre Olmesartan atherosclerosis Regression Evaluation (MORE) study was a double-blind trial in patients with hypertension at increased cardiovascular risk with carotid wall thickening and a defined atherosclerotic plaque that used non-invasive 2- and 3-dimensionaL (D) ultrasound (US), to compare the effects of a 2-year treatment based on either olmesartan medoxomil or atenolol on common carotid (CC) intima-media thickness (IMT) and plaque volume (PV). METHODS: A total of 165 patients (with systolic/diastolic blood pressure 140-180/ 90-105 mmHg) were randomized to receive either olmesartan (20-40 mg/day) or atenolol (50-100 mg/day). US was performed at baseline and 28, 52 and 104 weeks. The primary efficacy outcome was the change from baseline ( Delta) in CC IMT assessed by 2D US. Secondary outcomes included Delta PV assessed by 3D US and blood pressure (BP). RESULTS: Olmesartan and atenolo produced comparable significant reductions in CC-IMT; mean Delta IMT (SEM) was -0.090 (0.015) mm for oLmesartan and -0.082 (0.014) mm for atenolol. Mean Delta PV was -4.4 (2.3) microl and 0.1 (1.5) microl in the olmesartan and atenolol treated subjects, respectively, without significant between-treatment differences. In the subgroup of patients with baseLine PV > or = median (33.7 microl), significant between treatment differences existed in Delta PV (p = 0.023), because PV regressed significantly with olmesartan (Delta PV: -11.5 (4.4) microl) but not with atenolol ( Delta PV: 0.6 (2.5) microl). In these patients BP reductions were comparabLe. CONCLUSIONS: Carotid IMT and BP decreased similarly with olmesartan and atenolol, but only olmesartan reduced the volume of larger atherosclerotic plaques. PMID- 19124401 TI - New approaches to the assessment of left ventricular hypertrophy. AB - In hypertension, Left ventricular hypertrophy is initially a useful compensatory process that represents an adaptation to increased ventricular wall stress; however, it is also the first step toward the development of overt clinical disease. For this reason most international guidelines recommend the assessment of cardiac target organ damage in hypertensive patients for cardiovascular risk stratification. It is therefore of great importance to keep in mind the strengths and weakness of the different available methods for LVH assessment.Several methods are currently available for the assessment of LVH; however the various techniques differ in cost, availability, sensitivity and specificity. Due to its wide availability and its low cost, eLectrocardiography should be part of all routine assessment of subjects with high blood pressure; however, despite its good specificity, the sensitivity for LVH detection is low. Several other methods have been proposed for LVH detection. Cardiac magnetic resonance imaging allows 3D reconstruction of the heart with high spatial resolution; however its main limitation is represented by the relatively low availability and by its costs. Echocardiography certainly represents a valuable method for the detection of LVH in hypertensive patients, due to its wide availability and its relatively low cost. The main limitations of the technique are represented by the lower spatial resolution and reproducibility in comparison with magnetic resonance. The development of new matrix-array transducers and new software for 3D reconstruction with echocardiography make this approach particularly promising for the future; in the meantime, standard echocardiography, widely available and with low cost, will probably remain the most used tool for the evaluation of left ventricular structure and function in hypertension. PMID- 19124402 TI - Vascular remodeling: implications for small artery function and target organ damage. AB - At the level of the small artery, essential hypertension is associated with eutrophic inward remodeling. This involves reduction in lumen diameter by an increase in wall thickness. Previously thought to involve either hypertrophy or hyperplasia of the vascular smooth muscle cells in the media, it is now felt to be mediated by a functional property of the wall: myogenic tone. This is the ability of an artery to contract in response to an increase in intraluminal pressure. This autoregulatory function is also vital to ensure stabilisation of distal capillary pressures and so prevent, or limit, organ damage. Indeed in any animal model studied, when myogenic autoregulation is affected, target organ damage ensues. We have also observed, in two studies, that when myogenic autoregulation is damaged in the context of hypertension, eutrophic remodeling is replaced by an outward growth of the arterial wall with preservation of lumen diameter. This is called hypertrophic remodeling and, independently, has been observed by a number of groups in small arteries from patients with type 2 diabetes. We believe that this is a key reason for the unique propensity to hypertensive injury seen in patients with diabetes. We also discuss the significance of integrins, transmembrane proteins with wide ranging functions; from initiation of cell migration to intracellular signalling. Two particular integrins, alpha5beta1 and alphanubeta3, have been found to be necessary for both normal myogenic autoregulation and eutrophic remodeling and the possibility that damage to these may occur in diabetes is examined. PMID- 19124403 TI - The new European Society of Hypertension/European Society of Cardiology (ESH/ESC) Guidelines. PMID- 19124404 TI - Pulse pressure, arterial stiffness and wave reflections (augmentation index) as cardiovascular risk factors in hypertension. AB - For many years, the cyclic blood pressure (BP) curve was described exclusively from two specific points of this curve, the highest, called systolic blood pressure, and the lowest, called diastolic blood pressure, both dominating the basis of cardiovascular (CV) hypertensive epidemiology. Nowadays, it is largely admitted that the BP curve should be characterized from pulsatile arterial hemodynamics, thus leading to the definition of novel indices in CV epidemiology. The present chapter details these new aspects. PMID- 19124405 TI - Role of the augmentation index in hypertension. AB - Recent studies have revealed the clinical usefulness of central blood pressure (BP) as an index of risk for cardiovascular disease. The arterial pulse waveform is the sum of the forward pressure wave generated by left ventricular ejection and a backward propagating wave that is subsequently reflected from the peripheral site, and the time point at which these forward and backward propagating waves merge and the amplitude of the reflected (backward) wave affect the level of central BP. The augmentation index (AIx) has been proposed as a measure of the wave reflection, and its clinical usefulness has also been evaluated. In the process, the non-linear relationship between age and AIx, the prognostic value of AIx, and the various effects of antihypertensive drugs on AIx have been shown. However, the clinical usefulness of AIx has not been established, and several questions about its use remain. Future studies will be needed to address these questions, and may contribute to important changes in the management of cardiovascular disease. In this review, we present recent findings on the AIx and discuss the role of this parameter in clinical practice. PMID- 19124406 TI - Aspirin and Diabetes Mellitus: revisiting an old player. AB - Type 2 Diabetes Mellitus confers an excess risk of cardiovascular disease. The mechanisms involved in the development of the disease are an active field of research, and prompt the development of newer and safer therapeutics with implications for cardiovascular disease. Currently there is increasing awareness of the role of platelet dysfunction, low-grade chronic inflammation and thrombogenesis in the pathophysiology of insulin resistance, T2DM, as well as type 1 diabetes mellitus and cardiovascular disease. This new evolving knowledge has allowed a better understanding of the role of aspirin, an old medication with proven beneficial effects on patients with established cardiovascular disease. The influence of salicylates on insulin resistance, glucose homeostasis, platelet function and inflammatory pathways, in particular related to the activation of the NFkappaB pathway, is a promising field of active research, and will help in the management of both diabetes mellitus and atherosclerotic-related cardiovascular disease. PMID- 19124407 TI - The safety of drug-eluting stents. AB - Drug-eluting stents (DES) in percutaneous coronary interventions significantly reduce rates of restenosis and the need for new revascularizations compared with bare metal stents. However, as the use of DES has increased dramatically, questions have been raised about their long-term safety. Concerns about an increased risk of late stent thrombosis, particularly beyond the first year of treatment, have arisen and have been exacerbated by sparse and conflicting information, and boosted an intense debate between cardiologists. In this article, we reviewed the most recent information to clarify the conundrum of late stent thrombosis and the long-term safety of DES. PMID- 19124408 TI - Adverse cardiovascular effects of NSAIDs: driven by blood pressure, or edema? AB - The non-selective non-steroidal anti-inflammatory drugs (nsNSAIDs) and cyclooxygenase-2 (COX-2) inhibitors are commonly utilized agents for musculoskeletal conditions. The harmful cardiorenal effects of some nsNSAIDs are well described and thought to be related to inhibition of prostanoid synthesis. Since the non-specific inhibition of both cyclooxygenase enzymes was associated with a higher incidence of gastrointestinal side effects, the selective targeting of the COX-2 enzymes with the COX-2 inhibitors promised and delivered a lower incidence of gastrointestinal side effects. However, the COX-2 inhibitors have not been found to be bereft of cardiorenal side effects. Indeed, some of these agents lead to increased blood pressure, an excessive risk of congestive heart failure and pro-thrombotic effects, especially in high risk populations. These deletrious effects, however, may not be class-specific and possibly related to pharmacokinetics, enzyme specificity and endothelium effects. This article also reviews the body of literature linking the nsNSAIDs and COX-2 inhibitors with important adverse cardiorenal effects and their putative mechanisms. PMID- 19124409 TI - Of gender, statins, and coronary artery interventions. PMID- 19124410 TI - On the selective inhibitors of Cyclooxygenase-2: do we have a last word? PMID- 19124412 TI - Endothelial dysfunction: a potential tool in gender related cardiovascular disease. AB - The overwhelming importance of distinctive cardiovascular disease profile in women has stimulated enormous efforts to disclose its cause. In this review, we discuss vascular endothelium as a potential phenotypic marker for the genetic difference. As it is a potentially modifiable factor for cardiovascular disease, every effort should be made to detect it, either directly or indirectly, at the earliest in females who are at risk, so that the future cardiovascular events might be prevented. PMID- 19124411 TI - Capillary rarefaction in treated and untreated hypertensive subjects. AB - This study aimed to determine if capillary rarefaction is detectable and associated with endothelial dysfunction in persons with mild systolic blood pressure (SBP) elevation. Capillary density and endothelial function were quantified for 150 nondiabetic participants, grouped by blood pressure (BP) as normotensive, untreated high BP, and treated high BP. Structural capillary rarefaction measures were not different between the three groups. Functional capillary rarefaction measures were significantly lower in both high BP groups compared to normotensives, and correlated inversely with endothelial function. The study findings indicate that the hypertensive vascular pathologic process is already underway at modest levels of blood pressure elevation. PMID- 19124413 TI - Percutaneous coronary interventions and statins therapy. AB - Statins exert a number of beneficial effects on endothelial function and atherosclerotic plaque, modulating oxidative stress and inflammation, with subsequent, well documented, primary and secondary prevention of coronary artery disease. Periprocedural myocardial infarction and contrast induced nephropathy, after percutaneous coronary intervention (PCI), are associated with a worse outcome on long term follow-up. In the ARMYDA study, pretreatment with statins before elective PCI reduces periprocedural myocardial infarction in patients with stable angina. Moreover, the ARMYDA ACS was the first randomized, prospective trial that demonstrated that an acute loading with a high dose of atorvastatin prevents myocardial damage in patients with unstable syndromes undergoing early (<48 hours) coronary angiography and consequent angioplasty. Statins could also have beneficial effects by reducing expression of adhesion molecules in endothelial cells (ICAM-1 and E-Selectin) as demonstrated in the ARMYDA-CAMS study. Furthermore, patients receiving statins at the time of procedure show a significantly reduced incidence of contrast-induced nephropathy. All this evidence may strongly influence the clinical practice of an interventional cardiologist. PMID- 19124414 TI - Gender differences following percutaneous coronary intervention. AB - PCI is effective for reducing symptoms in patients with stable angina pectoris but does not improve prognosis. In earlier trials PCI has been associated with more procedure related complications in women than men, but this difference between genders has been less pronounced in more recent studies. In acute coronary syndromes there is no evidence of gender differences regarding the benefit of primary PCI for ST-segment elevation myocardial infarction. However, several trials of unstable angina and non-ST-segment elevation myocardial infarction indicate that women do not have the similar benefit of a routine, early, invasive treatment strategy compared with men. PMID- 19124415 TI - Endurance exercise and resistance training in cardiovascular disease. AB - Contrary to the age old taboo of exercise in cardiac patients, resistance training has been gaining importance recently as a safe, healthy fitness option in prevention of cardiovascular diseases, the leading killer disease in the population above 45 years in the United States. Endurance or aerobic exercise helps improve overall stamina and the ability of the heart to pump oxygenated blood in those with and without prior cardiovascular disease. In addition to modifying cardiovascular risks, resistance training has profound beneficial effects on improving muscle strength and endurance, preventing osteoporosis and improving quality of life both in the healthy and cardiovascular patients including women and heart failure patients. So resistance training should be regarded as a complementary fitness program rather that a substitute to endurance training. This review discusses the physiological phenomenon and benefits of exercise training programs on cardiovascular disease patients focusing on endurance exercise and resistance training. PMID- 19124416 TI - Local TAT-p27Kip1 fusion protein inhibits cell proliferation in rat carotid arteries. AB - INTRODUCTION: p27(Kip1) is a cyclin kinase inhibitor that induces cell cycle arrest. In this study, the efficacy of fusion protein TAT- p27(Kip1) to inhibit cell proliferation in rat perivascular injured carotid arteries was tested. METHODS: The cDNA of p27(Kip1) and GFP (green fluorescein protein) fused to the TAT epitope, which allows cell penetration, yielded TAT-p27 (Kip1) and TAT-GFP fusion proteins. In vitro biological activity on cell proliferation was evaluated by [(3)H] thymidine DNA incorporation in rabbit aortic endothelial cells (REC). An in vivo model used a silicone collar filled with saline positioned around the carotid vessel for 14 days to produce an increased adventitia cross-sectional area. RESULTS: TAT-p27(Kip1) inhibited REC proliferation in vitro using either 100, 200, and 500 nM compared to control (88.2 +/- 4.4, 81.3 +/- 7, 71.9 +/- 4.2 vs. 100 +/- 6.7%, N = 3, respectively, p < 0.05). This response was stable for purified proteins stored at -20*C for at least 23 days. In vivo , TAT-p27(Kip1) solution reduced adventitia cross-sectional area in a dose-dependent manner compared to TAT-GFP (area in mm(2) - TAT-p27(Kip1): 200 nM, 0.160 +/- 0.018; 500 nM, 0.050 +/- 0.005 vs. TAT-GFP: 500 nM, 0.595 +/- 0.066 vs. the contralateral: 0.047 +/- 0.005, N = 7, p < 0.01). CONCLUSION: Taken together, these results provide evidence that TAT-p27(Kip1) can inhibit vascular cells proliferation. It is the first successful demonstration that the cell permeable TAT-p27(Kip1) has potential as a vascular anti-proliferative agent. PMID- 19124417 TI - Avoiding restenosis: is there a role for glucocorticoids in the drug-eluting stent era? AB - Restenosis is an important limitation of percutaneous coronary interventions (PCI). In-stent restenosis is mainly due to neointimal hyperplasia, a proliferative process modulated by inflammatory mechanisms. Numerous technical and pharmacological means have been tested to reduce restenosis rates, with frequently disappointing clinical results. Drug-eluting stents (DES) have demonstrated a high efficacy in reducing restenosis, but there are some associated problems that limit its generalized utilization. Glucocorticoids (GC), as potent anti-inflammatory agents, may exert beneficial effects on neointimal proliferation. Clinical studies with oral and intracoronary GC therapy have demonstrated reduction in restenosis rates in selected patients. Although further investigations are warranted, GC might have a potential role for restenosis prevention in selected cases. PMID- 19124418 TI - Hypertension, a health economics perspective. AB - The economic aspects of hypertension are critical to modern medicine. The medical, economic, and human costs of untreated and inadequately controlled hypertension are enormous. Hypertension is distributed unequally and with iniquity in different countries and regions of the world. Treatment of hypertension requires an investment over many years to prolong disease-free quality years of life. The high prevalence and high cost of the disease impacts on the microeconomics and macroeconomics of countries and regions. The criteria used for inclusion in clinical guidelines for hypertension impact on the cost and cost/utility of diagnosis or treatment. PMID- 19124419 TI - Statins and stroke. AB - Statins play an important role in brain ischemia. These drugs reduce cholesterol levels, which have been related to a reduction in vascular event risk, but they also have other functions besides cholesterol metabolism, called pleiotropic effects. Statins play an important role during the acute phase of ischemia, and might have neuroprotective effects, as they act in several mechanisms during the acute phase of stroke, such as in nitric oxide (NO) and glutamate metabolism, inflammation, platelet aggregation, immune responses and apoptosis. They also have other functions that can be related, with better long-term outcome, to neurorepair mechanisms. Statins promote angiogenesis, endogenous cell proliferation, neurogenesis and new synapse formation. PMID- 19124420 TI - The therapeutic role of RAS blockade in chronic heart failure. AB - Cardiovascular disease represents a continuum that starts with risk factors such as hypertension and progresses to atherosclerosis, end-organ damage, and ultimately to chronic heart failure (CHF) and premature death. Renin-angiotensin system (RAS) blockade with angiotensin converting enzyme (ACE) inhibitors and/or angiotensin II type 1 receptor blockers (ARBs) has turned out to be beneficial at all stages of this continuum. Several mechanisms govern the progression of structural myocardial damage to end-stage CHF. Chronic neuroendocrine activation fosters left ventricular remodeling and dilatation and leads to clinical symptoms of CHF via forward/backward failure. RAS inhibition is a cornerstone of neuroendocrine blockade in CHF patients, and combined RAS blockade is especially effective in patients presenting with repetitive cardiac decompensations. This review focuses on the therapeutic role of inhibitors of different RAS components in chronic heart failure caused by systolic left ventricular dysfunction. PMID- 19124421 TI - PPARs as new therapeutic targets for the treatment of cerebral ischemia/reperfusion injury. AB - Stroke is a leading cause of death and long-term disability in industrialized countries. Despite advances in understanding its pathophysiology, little progress has been made in the treatment of stroke. The currently available therapies have proven to be highly unsatisfactory (except thrombolysis) and attempts are being made to identify and characterize signaling proteins which could be exploited to design novel therapeutic modalities. The peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that control lipid and glucose metabolism. PPARs regulate gene expression by binding with the retinoid X receptor (RXR) as a heterodimeric partner to specific DNA sequences, termed PPAR response elements. In addition, PPARs may modulate gene transcription also by directly interfering with other transcription factor pathways in a DNA binding independent manner. To date, three different PPAR isoforms, designated alpha, beta/delta, and gamma, have been identified. Recently, they have been found to play an important role for the pathogenesis of various disorders of the central nervous system and accumulating data suggest that PPARs may serve as potential targets for treating ischemic stroke. Activation of all PPAR isoforms, but especially of PPARgamma, was shown to prevent post-ischemic inflammation and neuronal damage in several in vitro and in vivo models, negatively regulating the expression of genes induced by ischemia/ reperfusion (I/R). This paper reviews the evidence and recent developments relating to the potential therapeutic effects of PPAR-agonists in the treatment of cerebral I/R injury. PMID- 19124422 TI - Endothelial progenitor cells: markers of vascular reparative capacity. AB - Assessment of the propensity for vascular events has been based on measurement of risk factors predisposing one to vascular injury. These assessments are based on the strong associations between risk factors such as hypertension, cholesterol levels, smoking, and diabetes which were first described almost a half century ago. The more recent discovery of the relationship between ongoing inflammation and clinical outcomes has led to a variety of blood-based assays which may impart additional knowledge about an individual's propensity for future cardiovascular events. Vascular health is now better represented as a balance between ongoing injury and resultant vascular repair, mediated at least in part by circulating endothelial progenitor cells. To date, one's risk for vascular events has focused exclusively on assessing propensity for vascular damage, either by assessing conventional risk factors which were initially identified over half a century ago, or more recently by assessing markers of inflammation and other circulating factors which area related to subsequent clinical events. Circulating endothelial progenitor cells play important roles in accelerating endothelialization at areas of vascular damage, and EPC enumeration is a viable strategy for assessing reparative capacity. To date, EPC numbers have been correlated with the numbers of cardiovascular risk factors, extent of coronary disease, and future cardiovascular events. Given that EPC enumeration and functional characterization represent the only assessment of the reparative side of the balance between damage and renovation, this technique may offer independent and different assessment of propensity to cardiovascular injury, greatly improving risk stratification of patients. PMID- 19124423 TI - Atrial fibrillation and renin-angiotensin system. AB - Atrial Fibrillation (AF) is one of the most frequent arrhythmias, especially in elderly patients. Cardiac overload increases the incidence of AF. Clinical presentation of atrial fibrillation can occur as nonsustained paroxysms, persistent episodes and in chronic-permanent form. The physio-pathological mechanisms are: *Circuit of multiple and anarchic re-entries *Atrial fibrillatory conduction *Re-entry circuit with fibrillatory conduction.Remodeling (electrical or structural) facilitates the appearance and persistence of AF: Neurovegetative changes and cytosolic Ca overload facilitate AF. Interstitial atrial fibrosis, in which Renin-Angiotensin System (RAS) hyperactivity is a main aspect of remodeling. There is clinical evidence that supports the antiatrial fibrillatory actions of RAS blockade. Potential mechanisms are: (a) direct modulation of ionic channels, (b) hemodynamic improvement, (c) reduction of atrial stretching, (d) antifibrotic effects. There is less clinical evidence with antialdosterone drugs, but theoretically these might also be useful. PMID- 19124424 TI - ACE-inhibitor, AT1-receptor-antagonist, or both? A clinical pharmacologist's perspective after publication of the results of ONTARGET. AB - Clinical Pharmacology is commonly accepted to be a bridging discipline between basic science observations and clinical practice. Today, it should be a major task of the clinical pharmacologist in academia to provide support in the interpretation of preclinical and clinical study data, to develop evidence-based treatment guidelines and to serve as drug expert supporting all disciplines of clinical medicine with specific pharmacological and therapeutic knowledge. The results of the ONTARGET-trial confront both researchers and clinicians with the unexpected truth that AT(1)-receptor-blockade with an angiotensin-receptor blocker (ARB) does not seem to have superior therapeutic benefit compared with an ACE-inhibitor (ACE-I) at reducing fatal and nonfatal cardiovascular events. The combination of the two drugs was associated with more adverse events without an increase in benefit. Therefore, the crucial question 'ACE-I, ARB, or both?' requires a new and critical appraisal depending on the medical indication for which these renin-angiotensin-system (RAS)-inhibitors are used: In a population of high-risk patients suffering from cardiovascular disease or diabetes mellitus, the evidence to favor an ARB over an ACE-I is still limited after ONTARGET and because of the higher costs for ARBs one can rather support the old therapeutic advice that ARBs are equally effective as ACE-Is and therefore therapeutic alternatives for patients with ACE-I intolerance. With respect to a very moderate additive BP-lowering effect of dual therapy with an ACE-I and an ARB seen in metaanalysis which was not even clearly attributable to dual RAS-inhibition and the increased adverse event rate in the combination treatment group of ONTARGET, this regimen seems not to be recommendable for the treatment of hypertension. Dual-RAS-blockade using an ACE-I-ARB-combination is an effective therapy to treat proteinuria and might be of therapeutic benefit especially in diabetic patients without concomitant diseases. There may be a therapeutic rationale to prefer ARBs over ACE-Is in well-selected patients with congestive heart failure (CHF) because a considerable amount of angiotensin II (Ang II) is produced independent of angiotensin-conversion-enzyme (ACE) in the failing heart and is therapeutically unaffected by ACE-I treatment. The results of the Val-HeFt and the CHARM-added study revealed additive effects of an ARB on heart failure related morbidity and mortality when added to existing therapy with an ACE-I suggesting a role for ACE I-ARB-combination treatment in well selected heart failure patients. Independent of the medical indication for its use, the concept of dual RAS-blockade with an ARB-ACE-I-combination should clinically be used with caution and a close monitoring of potassium levels and kidney function. Although the results of ONTARGET revealed equity of ramipril and telmisartan at reducing fatal and nonfatal cardiovascular events, we should not forget that pharmacologically not all ARBs are the same and the question if the study results of ONTARGET with telmisartan are transferable to the complete class of ARBs still merits further investigation. PMID- 19124426 TI - The role of the L-arginine-nitric oxide pathway in preeclampsia. AB - Preeclampsia (PE) is a major cause of maternal and perinatal mortality, especially in developing countries. Its etiology involves multiple factors, but no specific cause has been identified. Evidence suggests that clinical manifestations are caused by endothelial dysfunction. Nitric oxide (NO), which is synthesized from L-arginine in endothelial cells by the endothelial nitric oxide synthase (eNOS), provides a tonic dilator tone and regulates the adhesion of white blood cells and platelet aggregation. Alterations in the L-arginine-NO pathway have been associated with the development of PE. Various studies, reporting decreased, elevated or unchanged levels of nitrite (NO(2)) and nitrate (NO(3)), two end products of NO metabolism, have been published. Our group contributed to those contradictory reports describing cases of PE with both elevated and decreased levels of NO(2) and NO(3). Apparently, diminished levels of NO could be related to deficiencies in the ingestion of dietary calcium associated to low levels of plasma ionic calcium, which is crucial to the eNOS' activity. Also, low levels of NO could be associated with the presence of eNOS polymorphisms or the presence of increased levels of ADMA, the endogenous inhibitor of NO. High levels of NO associated to low levels of cGMP suggest a decreased bioactivity of NO, which is probably related to an increased degradation of NO caused by a high production of superoxide in states of infection and inflammation. The present article analyses and reviews the reported paradoxical roles of the L-arginine-NO pathway in PE and gives a possible explanation for these results. PMID- 19124425 TI - Preeclampsia and future cardiovascular risk: formal risk factor or failed stress test? AB - It is estimated that 10% of pregnancies are affected by hypertension worldwide. Approximately one-half of all hypertensive pregnancy disorders are due to preeclampsia, a pregnancy-specific disorder, its distinctive feature being either sudden onset, or worsening of pre-existing proteinuria. It has become increasingly recognized that women with a history of preeclampsia are at increased risk for future cardiovascular disease (CVD), but the mechanisms of this increase in risk are unclear. One possible explanation is that these two conditions share several common metabolic abnormalities as risk factors, including obesity, insulin resistance, and lipid abnormalities that may lead to preeclampsia and CVD at different times of a woman's life. Recent studies have revealed that, similar to CVD, several mediators of endothelial cell dysfunction are up-regulated in preeclampsia. Free radical derived oxidative stress, various inflammatory markers, including neutrophil response, C-reactive protein, and leukocyte adhesion, may contribute to endothelial dysfunction in both preeclampsia and coronary atherosclerosis. Alternatively, preeclampsia itself may induce metabolic and vascular changes that may increase the overall future risk for CVD in affected women. Therefore, at present, it remains unclear whether preeclampsia is a formal risk factor for CVD, or identifies women at increased risk for CVD later in life. Pending large-scale studies aiming to examine the causality of this association, women with a history of preeclampsia should be counseled regarding their increased risks for hypertension and other cardiovascular sequelae later in life, followed closely and treated aggressively for modifiable CVD risk factors. PMID- 19124427 TI - Does lowering cholesterol have an impact on the progression of aortic stenosis? AB - Several studies suggest that atherosclerotic disease is not a focal disease restricted to culprit lesions in the intima of the arterial wall, but seems to act as a general disease affecting the entire cardiovascular system. Evolving research has lately focused on the atherosclerotic component in calcific aortic stenosis (AS) as it seems that the valve is affected in a pattern similar to that of the vasculature. The hope is therefore, that we someday in the management of patients with calcific AS can apply some of the same treatment strategies as in atherosclerotic vascular disease. This article reviews the pathophysiological mechanisms of calcific AS, reviews current clinical trials of statin use in aortic stenosis and reports on on-going trials, evaluating whether cholesterol lowering therapy can slow disease progression in different populations. Finally, we review if computerized tomography, biomarkers, and clinical characteristics such as left ventricular ejection fraction, can be useful in stratifying patients to potential benefit of statin therapy. PMID- 19124429 TI - Prehypertension: should we be treating with pharmacologic therapy? AB - Prehypertension, defined by Seventh Joint National Committee (JNC 7) as a blood pressure (BP) 120-139/80-89 mm Hg, was controversial. Approximately 31-37% of US adults are prehypertensive, and approximately 12-14% have BP of 130-139/85-89 mm Hg or ;Stage 2' prehypertension, is associated with approximately 3-fold greater likelihood of developing hypertension and roughly twice the cardiovascular events than BP <120/80 mm Hg. Lifestyle change is the only intervention recommended for most prehypertensives. When fully implemented, lifestyle changes lower BP and prevent cardiovascular events, but evidence for community-wide effectiveness is limited. The Trial of Preventing Hypertension (TROPHY) documented that angiotensin receptor blockade safely lowers BP and prevents and/or delays hypertension in Stage 2 prehypertensives. Prehypertensives with diabetes or nephropathy are at high risk and should receive antihypertensive treatment according to JNC 7. Epidemiological data suggest that the number needed to treat to prevent a cardiovascular event in these at-risk Stage 2 prehypertensives is similar to Stage 1 hypertensives when both groups have one or more concomitant risk factors. Clinical trials are urgently needed to address this question. In the absence of clinical trials data, we believe it is prudent for the concerned clinician to consider initiating antihypertensive pharmacotherapy in selected Stage 2 prehypertensive patients at significant absolute risk for progression to hypertension and cardiovascular events. PMID- 19124428 TI - Stroke prevention: modifying risk factors. AB - Risk factor modification remains as the principal aspect of care for stroke prevention. Understanding of risk factors has advanced and several options are now available to treat modifiable risk factors. However, effective treatment remains a challenging task in clinical practice. Prevention begins with awareness of risk factors by patients and clinicians. Risk factor assessment along with overall stroke risk estimation should be part of evaluation of patients with stroke, and used with careful clinical judgment. In this review, we discuss the impact of modifiable traditional vascular risk factors on ischemic stroke, interventions for stroke prevention, and evidence for early treatment of risk factors where available, as well as areas of research progress. Emphasis should be put on the education of patients, the community, and medical personnel. Future research in the field of genetic determinants of vascular risk factors and stroke will increase our understanding of the underlying mechanisms of cerebrovascular disease and likely result in development of new therapies and individualized programs for stroke prevention. PMID- 19124430 TI - Prevention of coronary heart disease in women. AB - OBJECTIVE: Coronary heart disease (CHD) mortality is higher in women than in men and misdiagnosis of CHD in women is one of the reasons for this, with differences in the presentation of CHD between men and women being a cause for the misdiagnosis. This review discusses the need for evidence-based guidelines to diagnose and treat CHD in women. METHODS: Reviews, randomized controlled trials, and other studies pertinent to the topic were obtained using electronic search strategies, such as MEDLINE and Cochran Library, as well as manual selection. Sources selected were limited to those that discussed CHD, with specific emphasis placed on sources that focused on CHD in women. Selected studies were then assessed for quality of data and relevance via analysis of the study's methodology, results, and data. Results of selected studies were then stratified using a rating system devised to determine the quality of results using the scientific evidence provided for them. The references of the selected studies were then used to obtain and analyze additional studies in the same manner. RESULTS: Control of lifestyle factors such as smoking, physical activity, diet, and weight are all necessary in women to control CHD, as is the maintenance of healthy lipid levels and blood pressure. Angiotensin-converting enzyme inhibitors and antiplatelets can help aid lifestyle changes in CHD management for women while hormone therapy and vitamin E have no proven benefits in CHD management. CONCLUSIONS: New gender- and evidence-based guidelines for the prevention of CHD in women need to be developed and adopted by physicians so that prevention, diagnosis, and treatment of CHD is made more effective. PMID- 19124431 TI - Hypercholesterolemia-associated endothelial progenitor cell dysfunction. AB - Hypercholesterolemia has been associated with increased cardiovascular risk by contributing to mechanical endothelial injury and dysfunction. There is evidence that chronic exposure to increased plasma cholesterol levels might also impair the repair of lipoprotein-mediated endothelial injury, possibly by reducing the availability and function of circulating endothelial progenitors. This review summarizes current knowledge about the mechanisms of lipoprotein-mediated endothelial injury and endothelial progenitor cell assisted vascular repair; the influence of hypercholesterolemia on endothelial progenitor cell dysfunction will be also addressed. PMID- 19124432 TI - Cardiogenic potential of endothelial progenitor cells. AB - Transplantation of endothelial progenitor cells (EPCs) are one of the promising strategies to recover the capillary flow in ischaemic diseases such as ischaemic heart disease and peripheral artery disease (PAD) in the leg. However, our previous and another group's works suggested the scarcity of the number of EPCs in peripheral blood might cause insufficient effect for the ischaemic diseases. There are several strategies to overcome this issue, such as (1) in vivo EPC expansion; (2) ex vivo EPC expansion; (3) local (not systemic) EPC injection; and (4) modification of EPC by gene transfer. Recent publications from our own and other groups have reported the possibility of cardiogenic potential of EPCs. We would like to focus on the strategies of EPC transplantation and cardiomyogenesis of EPCs in this review. PMID- 19124434 TI - Genetics of diabetic nephropathy. AB - Genetic susceptibility has been proposed as an important factor for the development and progression of diabetic nephropathy, and research efforts have been invested worldwide to identify the susceptibility gene for diabetic nephropathy. Although, several candidate genes were shown to be associated with the disease, the results were not always consistent; most of the genes conferring susceptibility to diabetic nephropathy remain to be identified. Recent development of the single nucleotide polymorphism (SNP) typing technology and collation of information on linkage disequilibrium in the human genome have facilitated genome-wide association studies (GWASs) for investigating novel disease-susceptibility genes across the entire human genome. GWASs are considered a powerful and promising approach and are expected to be useful for identifying convincing susceptibility genes for several common diseases; however, to date, these studies have not been able to completely cover the entire human genome. PMID- 19124433 TI - Systemic and uteroplacental renin--angiotensin system in normal and pre-eclamptic pregnancies. AB - Pregnancy is characterized by an increase in many of the different components of the circulating renin-angiotensin system (RAS). However, the physiological mechanisms of stimulated RAS activity during pregnancy are unknown. Even less understood is how this system may be altered in pre-eclampsia, a hypertensive disorder of pregnancy. Additional studies have shown the presence of a local tissue specific RAS in the uteroplacental unit of normal and pre-eclamptic pregnancies. Differences in normal pregnant and pre-eclamptic RAS component regulation may provide insight into the mechanisms responsible for the clinical pathological features of pre-eclampsia. Specifically, this review summarizes the key findings in the circulating and uteroplacental RAS in normal and pre eclamptic pregnancies. PMID- 19124435 TI - New techniques for assessment of vascular function. AB - The noninvasive measurement of hemodynamic variables associated with hypertension and cardiovascular disease processes needs to be recognized as a viable adjunct to clinical practice. This review traces the history of the inception and development of noninvasive measurement of hemodynamic variable. It then identifies well established, useful, and available devices, and then notes clinical studies verifying the clinical relevance of these measurements. Given the need to intervene earlier in the course of cardiovascular disease processes, tools are needed to assist the medical team to evaluate, prognosticate, and guide their patient's therapy correctly. It is the goal of this review to heighten the awareness and enhance and encourage the implementation of these devices in our armamentarium for the betterment of our patient's health. PMID- 19124436 TI - Recurrent stroke: where do we stand with the secondary prevention of noncardioembolic ischaemic strokes? AB - Strokes recur in 6-20% of the patients, most commonly within the first year; after a TIA or minor stroke; most recurrences will occur within the first 90 days. Our ability to identify patients at high risk is poor and most recurrent strokes cannot be explained by traditional risk factors. In 30-45% of the cases the second stroke will be of a different subtype. Moreover, patients are faced with other risks, like cardiac events and cognitive decline. With the population aging, the need for timely and effective secondary prevention strategies is more pressing than ever. This paper summarizes recent advances in pharmacological secondary prevention after a non-cardioembolic ischaemic stroke, and highlights critical questions still in need of answers. PMID- 19124437 TI - Questions to ponder. PMID- 19124438 TI - Elevated infection parameters and infection symptoms predict an acute coronary event. AB - BACKGROUND: The etiology and significance of flu-like symptoms often appearing before myocardial infarction should be clarified. METHODS: In a case-control study of 323 matched controls and a random sample of 110 out of 351 cases the presence of infection symptoms during the preceding four weeks before admission were asked and blood samples taken. RESULTS: Enterovirus (EV), herpes simplex virus (HSV), and Chlamydia pneumoniae IgA titers were significantly higher in cases than in controls (p<0.001, 0.008 and 0.046, respectively). Flu-like symptoms appeared significantly more often in patients than in controls the most common one being fatigue (p<0.001). In controls with fatigue, EV and HSV titers showed a trend to be higher (1.50 vs 1.45 and 4.29 vs 3.73) than in controls without fatigue but only HSV titers were statistically significantly higher (3.47 vs 3.96, p = 0.02). Even CRP and amyloid A concentrations (3.49 vs 2.08, p<0.0001 and 5.70 vs 3.77 mg/l, p = 0.003, respectively) as well as C4 (0.40 vs 0.44, p = 0.02) were higher in controls with fatigue. CONCLUSIONS: Odds ratios for a coronary event in a logistic regression model were 4.79 for fatigue and 2.72 for EV antibody levels in their fourth quartile. A linear-by-linear association test showed increasing number of single symptoms with higher EV titer quartiles (p = 0.004). PMID- 19124439 TI - Cardiovascular disease in chronic kidney disease. AB - The presence of kidney disease, manifested by low glomerular filtration rates (GFR) and/or large amounts of protein in the urine, is independently associated with increased rates of cardiovascular disease (CVD). The severity of kidney disease is associated with graded increases in risk for CVD and death. Chronic kidney disease (CKD) should be recognized and treatment initiated early to maximize the chances for slowing nephropathy progression and reducing proteinuria. We recommend screening for CKD in all patients with CVD, including computing an estimated GFR and evaluating for proteinuria using a spot urine albumin:creatinine ratio. Aggressive management of traditional cardiovascular risk factors should be employed in this high-risk population, specifically rigorous hypertension control (including the use of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blocking agents (ARB)), management of hyperglycemia, hyperlipidemia and smoking cessation. Further studies are needed to identify the unique renal failure-related (non-traditional) risk factors that contribute to accelerated atherosclerosis in this population and performance of randomized trials to assess the effects of cardiovascular interventions in individuals with CKD. PMID- 19124440 TI - Thiazolidinedione associated volume overload and pulmonary hypertension. AB - Thiazolidinediones are currently indicated for the treatment of Type 2 Diabetes. This class of drugs has been associated with several adverse reactions associated with volume overload. This report describes a case of a 65 year old African American female with a history of hypertension and obesity, and taking rosiglitazone (Avandia) for her Type 2 Diabetes whose evaluation for chest pain resulted in the incidental finding of pulmonary hypertension noted on echocardiogram. The subsequent evaluation, follow-up and treatment are discussed along with potential pitfalls and implications for clinical care. PMID- 19124441 TI - Pathophysiology, diagnosis, and management of aortic dissection. AB - Aortic dissection is an uncommon but potentially fatal disease with catastrophic complications. A high level of suspicion is required for successful diagnosis as presenting symptoms are so variable that dissection may be overlooked in up to 39% of cases. It most commonly presents in the elderly population with a history of chronic hypertension. Rapid intervention is necessary as delay leads to higher mortality. Despite advances in diagnostic and therapeutic techniques, morbidity and mortality remains high. Advances in diagnostic imaging have raised the awareness of variants of aortic dissection, including intramural hemorrhage and penetrating aortic ulcer. This distinction is important as the clinical course of these variants differs from that of classical aortic dissection, and thus treatment may also differ. Understanding of these variants has also led to the recognition of markers that may help predict progression to classical aortic dissection and thus warrant closer vigilance in selected patient populations. The recognition that rapid diagnosis is required for management of aortic dissection has led to the investigation of serum tests as diagnostic aids. Serum smooth muscle myosin heavy chain, d-dimer, and serum soluble elastin fragments are promising tests that may help raise suspicion for the diagnosis of acute aortic dissection. The high mortality associated with surgical therapy has led to investigation of alternative approaches. Endovascular therapy has emerged as a viable option in patients with type B dissection who are too unstable for surgery. However, long-term follow up is required to validate this procedure. PMID- 19124443 TI - Monitoring oral antiplatelet therapy: is it justified? AB - Platelets play a key role in the initial formation and progression of intravascular thrombus. During coronary and peripheral vascular interventions, antiplatelet therapy is used to impair platelet reactivity in order to minimize adverse ischemic events. Chronic antiplatelet therapy is also administered to decrease the long term risk of major adverse cardiovascular events in patients with high atherothrombotic burden. However, due to a heterogeneous response to antiplatelet agents, a substantial number of patients with cardiovascular disease remain at risk despite adherence to therapy. Since the availability of point-of care rapid platelet function assays, quick and reproducible evaluation of platelet function after the administration of aspirin, thienopyridines, and glycoprotein IIb/IIIa inhibitors is possible. Various definitions of suboptimal platelet inhibition with oral antiplatelet therapy and associated risk factors are presented here. An algorithm to guide optimal antiplatelet therapy based on rapid platelet function testing during cardiovascular interventions is also presented. PMID- 19124444 TI - Mechanisms of alcoholic heart disease. AB - Compromised heart function is regularly seen in patients with chronic alcohol ingestion and is often manifested as cardiomegaly, reduced myocardial contractility (with concomitant reductions in ejection fraction and stroke volume), myocardial fibrosis, enhanced risk of stroke and hypertension, and disruptions in the myofibrillary structure. A number of mechanisms including oxidative damage, deposition of triglycerides, altered fatty acid extraction, decreased myofilament Ca(2+) sensitivity, and impaired protein synthesis have been proposed for the development of alcoholic cardiomyopathy. Nonetheless, the underlying mechanism(s) has not been delineated. Several alcohol metabolites have been identified as specific toxins of myocardial tissue, including ethanol, its first and major metabolic product--acetaldehyde--and fatty acid ethyl esters. Acetaldehyde directly impairs cardiac contractile function, disrupts cardiac excitation-contraction coupling and promotes oxidative damage and lipid peroxidation. Unfortunately, the most direct approach to studying this (direct administration of acetaldehyde) is impossible, since direct intake of acetaldehyde is highly toxic and unsuitable for chronic studies. In order to overcome this obstacle, transgenic mice have recently been produced to artificially alter ethanol/acetaldehyde metabolism, resulting in elevated acetaldehyde levels after ethanol ingestion. This review will summarize some of the postulated mechanisms for alcoholic cardiomyopathy, with special emphasis on animal models. PMID- 19124442 TI - Ethanol for cardiac ischemia: the role of protein kinase c. AB - The physiological effects of ethanol are dependent upon the amount and duration of consumption. Chronic excessive consumption can lead to diseases such as liver cirrhosis, and cardiac arrhythmias, while chronic moderate consumption can have therapeutic effects on the cardiovascular system. Recently, it has also been observed that acute administration of ethanol to animals prior to an ischemic event provides significant protection to the heart. This review focuses on the different modalities of chronic vs. acute ethanol consumption and discusses recent evidence for a protective effect of acute ethanol exposure and the possible use of ethanol as a therapeutic agent. PMID- 19124445 TI - The left atrium, atrial fibrillation, and the risk of stroke in hypertensive patients with left ventricular hypertrophy. AB - The Losartan Intervention For Endpoint reduction in hypertension (LIFE) study provided extensive data on predisposing factors, consequences, and prevention of atrial fibrillation (AF) in patients with hypertension and left ventricular (LV) hypertrophy. Randomized losartan-based treatment was superior to atenolol-based treatment for reducing new-onset AF and complications, especially stroke, associated with new-onset or pre-existing AF. Potential mechanisms of AF prevention by angiotensin receptor blockade supported by LIFE results include greater reduction in left atrial size and LV hypertrophy. Differential effects of antihypertensive treatment on the left atrium and left ventricle may help prevent AF and reduce risk of stroke associated with hypertensive heart disease. PMID- 19124446 TI - Prospective study of ambulation after open and laparoscopic colorectal resection. AB - PURPOSE: Open and laparoscopic surgical approaches each have specific advantages. This study compares ambulation, hospital length of stay (LOS), and incision length after open and laparoscopic colorectal resection. METHODS: All consecutive patients undergoing colorectal resection over a 2 year period ending August 2002 were followed prospectively. Ambulation, LOS, and incision length were recorded. Hybrid low anterior resection (LAR) patients had laparoscopic splenic flexure takedown, vessel ligation, and proximal rectal mobilization followed by planned inferior laparotomy to complete the case. Groups were compared using Student's t test. RESULTS: Equivalent open and laparoscopic groups were comparable in terms of gender, age, body mass index, ASA class, indication for operation, and resection performed. Seventy open colectomy patients were compared with 99 laparoscopic-assisted colectomy patients. On average, patients in the open and laparoscopic groups ambulated 67 and 390 feet, respectively, on postoperative day 1 (P < .001), 290 and 752 feet on day 2 (P < .001), and 495 and 965 feet on day 3 (P < .001). The average LOS in the open group was 9.3 days compared with 5.9 days in the laparoscopic group (P < .001). The average incision length in the open group was 19.7 cm compared with 5.3 cm in the laparoscopic group (P < .001). Seventeen open LAR patients were compared with 30 hybrid LAR patients. On average, patients in the open and hybrid groups ambulated 22 and 150 feet, respectively, on postoperative day 1 (P = .003), 105 and 433 feet on day 2 (P = .003), and 369 and 488 feet on day 3 (P = .43). The average LOS in the open group was 10 days compared with 8.5 days in the hybrid group (P = .46). The average incision length in the open group was 19.8 cm compared with 10.8 cm in the hybrid group (P < .001). When all 216 patients were considered, the 91 patients with incisions shorter than 8 cm (average 4.6 cm) ambulated 396, 752, and 956 feet on consecutive days whereas the 125 patients with incisions 8 cm or longer (average 16.9 cm, P < .001) ambulated 101, 334, and 521 feet on consecutive days (all P values <.001). Average LOS in the <8-cm group was 6 days compared with 8.9 days in the > or =8-cm group (P < .001). CONCLUSIONS: Patients undergoing minimal access colorectal surgery ambulated significantly further than equivalent open patients in the early postoperative period and had a shorter LOS. PMID- 19124447 TI - Aging and the laparoscopic revolution: has the landscape changed? PMID- 19124448 TI - 120-day comparative analysis of adhesion grade and quantity, mesh contraction, and tissue response to a novel omega-3 fatty acid bioabsorbable barrier macroporous mesh after intraperitoneal placement. AB - PURPOSE: This study aimed to evaluate adhesion formation, mesh contraction, and tissue response to an omega-3 fatty acid barrier-coated lightweight polypropylene mesh (C-Qur) after intra-abdominal placement, and compare these properties to those of other commercially available meshes. MATERIALS AND METHODS: After randomization, 3 x 3 cm pieces of Atrium C-Qur, Mesh ProLite Ultra, Composix, Parietex, Proceed, Sepramesh, and DualMesh were sewn to the intact peritoneum on either side of a midline incision in 41 New Zealand white rabbits. Necropsy was performed at 120 days, and explants were evaluated for adhesion grade, adhesion amount, and mesh contraction. Histologic evaluation included extent of capsule formation, abdominal wall tissue ingrowth, degrees of inflammation and vascularization of the surrounding tissue, and the presence of mesothelialization. Results. There were no significant differences between the C Qur mesh and the commercially available meshes tested with regard to adhesion grade or amount, although percentage adhesion coverage for the C-Qur mesh was much less than for Composix and Proceed. The C-Qur mesh contracted less than all meshes, significantly less (P < .05) than DualMesh or Proceed. DualMesh exhibited the greatest amount of capsule formation and inflammation on its parietal side as compared with the other meshes. CONCLUSIONS: Placing lightweight polypropylene mesh with an omega-3 fatty acid barrier coating intraperitoneally results in more favorable adhesion characteristics compared with Composix and Proceed meshes at 120-day explantation after intraperitoneal placement. The minimal amount of contraction and favorable tissue response in comparison to other commercially available meshes makes C-Qur mesh a practical alternative for laparoscopic and open ventral hernia repair. PMID- 19124449 TI - Basal cell carcinoma of the buccal mucosa in a patient with nevoid basal cell carcinoma syndrome. AB - Basal cell carcinoma (BCC) of the oral cavity is a controversial lesion with clinical and histopathologic features that overlap with those of peripheral ameloblastoma (PA). Ber-EP4, a cell surface glycoprotein preferentially expressed in BCC of the skin, has been suggested as a useful marker to support the diagnosis of oral BCC.This study presents a case of intraoral BCC arising in the anterior buccal mucosa of a patient with nevoid basal cell carcinoma syndrome (NBCCS), which represents a previously unreported clinical finding, to our knowledge. Histopathologic and immunohistochemical features of the case were compared to examples of PA, conventional intraosseous ameloblastoma, sporadic cutaneous BCC and cutaneous BCC from NBCCS patients. Ber-EP4 expression by the oral tumor was distinct from both peripheral and intraosseous forms of ameloblastoma and was identical to cutaneous BCC in both sporadic and syndromic settings. PMID- 19124450 TI - Clinicopathologic analysis of 4 perivascular epithelioid cell tumors (PEComas) of the gastrointestinal tract. AB - This study describes the clinical, histologic, and immunohistochemical features of four cases of GI PEComa. The size of the tumors ranged from 3.5 to 6.0cm in diameter, and all were located in wall of the large bowel. Microscopically, the tumors were characterized by an epithelioid arrangement of tumor cells, which had abundant clear to pale eosinophilic granular cytoplasm, vesicular nuclei and prominent nucleoli. The stroma was rich in capillaries, a sinusoidal vasculature and thick-walled blood vessels. Mitotic figures were generally rare. Immunohistochemically, the tumors were positive for Vimentin (4/4), HMB-45 (4/4), Melan-A(4/4),alpha-smooth muscle actin (4/4), and desmin (3/4). The patients have been well with no evidence of disease at 8, 15, 32 and 36 months,respectively, after the surgical operation. GI tract PEComas should be regarded as tumors of an uncertain malignant potential until long-term outcome data for a larger number of patients become available. PMID- 19124451 TI - Immunohistochemical expression of the human sodium/iodide symporter distinguishes malignant from benign gastric lesions. AB - AIM: Sodium/iodide symporter (NIS) is a transmembrane protein that mediates the transport of I(-). The aim was to evaluate the immunohistochemical expression of the human homolog of NIS (hNIS) in a wide spectrum of gastric lesions. MATERIALS AND METHODS: Seventy-seven samples were stained immunohistochemically with a monoclonal antibody for hNIS, including 14 with normal gastric mucosa, 14 with chronic atrophic gastritis with foveolar hyperplasia, 15 with chronic atrophic gastritis with intestinal metaplasia, 6 with chronic atrophic gastritis with atypical regenerative hyperplasia, 8 with chronic atrophic gastritis with dysplasia, 15 with invasive adenocarcinoma, 3 with well-differentiated neuroendocrine tumor, and 2 with gastrointestinal stromal tumors (GISTs). RESULTS: hNIS stained the basolateral cytoplasmic portion of foveolae in normal mucosa, in 13 cases of chronic atrophic gastritis with foveolar hyperplasia, and in only 1 case of regenerative atypical hyperplasia. hNIS was consistently absent in intestinal metaplasia, in dysplastic glands, and in the cells constituting invasive carcinoma, well-differentiated neuroendocrine tumors, and GIST. CONCLUSION: It seems that lack of hNIS can be useful in distinguishing foveolar hyperplasia from dysplastic glands. PMID- 19124452 TI - Review article: pulmonary sarcomatoid carcinomas: a practical overview. AB - Pulmonary sarcomatoid carcinomas (PSCs) are currently defined as poorly differentiated non-small-cell carcinomas containing a component with sarcoma or sarcoma-like (spindle and/or giant cell) features. They consist of 5 major histological variants, namely pleomorphic carcinoma, spindle cell carcinoma, giant cell carcinoma, carcinosarcoma, and pulmonary blastoma. The segregation of PSCs into a distinct clinicopathologic entity seems justified on the basis of morphologic, behavioral, and genotypic/phenotypic attributes. As a group, PSCs generally run an aggressive clinical course and may cause major difficulties in the differential diagnosis with other primary and secondary malignancies of the lung. At present, PSCs are believed to represent a family of carcinomas "in transition," in which diverse pathways of clonal evolution account for histological differences of a common ancestor lesion. The sarcomatous or sarcomatoid component of these tumors is thought to derive from carcinoma cells during the progression of carcinogenesis through the activation of an epithelial mesenchymal transition program leading to sarcomatous transformation or metaplasia (conversion paradigm). Conceivably, targeting the epithelial mesenchymal transition program could become a valid therapeutic strategy for these life-threatening tumors, whose sensitivity to current medical manipulation is disappointing. PMID- 19124453 TI - Delayed enhancement morphology on cardiac magnetic resonance imaging is correlated with signal-averaged electrocardiogram and QT dispersion in myocardial infarction. AB - BACKGROUND: In patients with myocardial infarction, ventricular tachycardia is related with nonconductive ventricular scar. Cardiac magnetic resonance imaging is an excellent modality to evaluate myocardial scars in myocardial infarction. Furthermore, late potential obtained from signal-averaged electrocardiogram and QT dispersion are both well-known parameters for predicting lethal arrhythmias. METHODS AND RESULTS: To investigate whether the pattern of necrotic scar tissue visualized by delayed enhancement on cardiac magnetic resonance imaging is associated with late potential and QT dispersion, we measured late potential and QT dispersion in 27 patients (68 +/- 8 years old) with a prior myocardial infarction. Cardiac magnetic resonance imaging was also obtained using a 1.5 tesla cardiac magnetic resonance scanner, and delayed enhancement was analyzed in the short axis of the left ventricle. By conducting this, we tried to determine whether the pattern of necrotic scar tissue predicts lethal ventricular arrhythmias. Semiquantitative patchy scores were identified as the mean patchy score and the maximum patchy score in each patient. There were 9 patients with a positive late potential and 18 patients with a negative late potential. Patients with positive late potentials had significantly larger mean (1.7 +/- 0.3) and maximum (2.2 +/- 0.6) patchy scores than patients with negative late potentials (mean, 1.3 +/- 0.2, P < .05; maximum, 1.7 +/- 0.4, P < .05). QT dispersion was significantly correlated with the number of slices showing delayed enhancement, which reflects the size of necrotic scar tissue (r = .59, P < .05). CONCLUSIONS: These findings suggest that the pattern of necrotic scar tissue visualized by delayed enhancement with cardiac magnetic resonance imaging was correlated to the predictive indices of lethal ventricular arrhythmias. PMID- 19124454 TI - Angio-Seal use in patients with peripheral arterial disease (ASPIRE). AB - PURPOSE: To investigate the incidence of complications after the use of an arterial closure device (Angio-Seal) in patients with peripheral arterial disease. METHODS: In 105 consecutive patients after transfemoral catheterization, the puncture site was closed using a closure device (Angio-Seal). Colour-flow duplexsonography studies were conducted 1 to 4 days before, within 3 days after and 3 month after the intervention. RESULTS: All patients had peripheral arterial disease, 34 had calcification at the puncture site. Detection of calcification did not prevent device deployment. Complications (2 minor bleedings, 1 pseudoaneurysm) were not associated with high risk groups (these were: 69 antegrade punctures, 22 obese and 32 hypertensive patients). Three-month postinterventional diameter and blood velocity changes were <1%. CONCLUSIONS: Patients with peripheral arterial disease in the region of the puncture site and patients at higher complication risk can safely and effectively be closed with an Angio-Seal device. At the puncture site, no lumen change can be observed 3 months postinterventional. PMID- 19124455 TI - Protein Z G79A polymorphism in patients with severe sepsis. AB - The aim of the study is to investigate whether the presence of a protein Z polymorphism is a risk factor for the development and outcome of sepsis. Sepsis is a clinical syndrome characterized by the presence of systemic signs and symptoms of inflammation. When sepsis leads to organ failure, the term severe sepsis and septic shock is used. The genetic causes of severe sepsis are not fully explained. Protein Z is a vitamin K- dependent glycoprotein and a member of the coagulation cascade. The study included 53 patients with severe sepsis and 70 control healthy volunteers without a familial history of thrombosis. The G79A polymorphism of intron F of the protein Z gene was analyzed by the method of polymerase chain reaction-based DNA analysis. The protein Z intron F G79A polymorphism frequencies of the patients and controls were 43.4% and 40%, respectively. Carrying 79 AA genotype could be a risk factor for severe sepsis and septic shock (OR = 4.5, 95% CI: 0.45-46.1), but it could not find any difference between survivor and nonsurvivor groups. They concluded that the frequency of intron F G79A polymorphism of protein Z gene was higher in patients than controls, and carrying 79 AA genotype could be a risk factor for severe sepsis and septic shock. PMID- 19124456 TI - Protein tyrosine kinase, syk: a key player in phagocytic cells. AB - Spleen tyrosine kinase (Syk) is a non-receptor protein tyrosine kinase expressed in a wide range of haematopoietic cells. At the initial stage of investigation, main exploring was toward its functions in platelets and in classical immunoreceptor signalling. However, Syk has now been recognized as a key player in both innate and adaptive immunity. Especially, in phagocytosis, Syk plays essential roles in signalling evoked by various types of receptors such as FcgammaR, CR3, Dectin-1 and apoptotic cell-recognizing receptor. A variety of upstream immunoreceptor tyrosine-based activation motif-like molecules have been found and are still in the course of new studies. On the contrary, downstream effectors to explain diverse function of Syk are still under exploration. As its novel function, we propose the role of Syk in the regulation of alpha-tubulin acetylation. Further investigation on the effectors of Syk would give us more information in relation to therapeutic molecular targets. PMID- 19124457 TI - Highly active low magnesium hammerhead ribozyme. AB - Hammerhead (HH) ribozymes can be used for highly specific inhibition of gene expression through the degradation of target mRNA. In vitro experiments with minimal HH domains demonstrated that the efficiency of catalysis is highly dependent on concentration of magnesium ions. Optimal ion requirements for HH catalysed RNA cleavage are far from these found in the cell. Recently, it has been proposed that the efficiency of HH ribozymes can be increased at low magnesium concentration through stabilization of a catalytically active conformation by tertiary interactions between helices I and II. We designed a ribozyme stabilized by GAAA tetraloop and its receptor motifs and demonstrated that it can efficiently catalyse target RNA hydrolysis at submillimolar Mg(2+) concentrations in vitro as well as in cultured cells. Both unmodified and locked nucleic acid-modified extended ribozymes proved superior to the minimal core ribozyme and DNAzyme against the same target sequence. PMID- 19124458 TI - Systematic comparison of in vitro motile properties between Chlamydomonas wild type and mutant outer arm dyneins each lacking one of the three heavy chains. AB - Outer arm dynein (OAD) of cilia and flagella contains two or three distinct heavy chains, each having a motor function. To elucidate their functional difference, we compared the in vitro motile properties of Chlamydomonas wild-type OAD containing the alpha, beta, and gamma heavy chains and three kinds of mutant OADs, each lacking one of the three heavy chains. For systematic comparison, a method was developed to introduce a biotin tag into a subunit, LC2, which served as the specific anchoring site on an avidin-coated glass surface. Wild-type OAD displayed microtubule gliding in the presence of ATP and ADP, with a maximal velocity of 5.0 mum/s, which is approximately 1/4 of the microtubule sliding velocity in the axoneme. The duty ratio was estimated to be as low as 0.08. The absence of the beta heavy chain lowered both the gliding velocity and ATPase activity, whereas the absence of the gamma heavy chain increased both activities. Strikingly, the absence of the alpha heavy chain lowered the gliding velocity but increased the ATPase activity. Thus, the three heavy chains are likely to play distinct roles and regulate each other to achieve coordinated force production. PMID- 19124459 TI - Analysis of transient and catalytic desosamine-binding pockets in cytochrome P 450 PikC from Streptomyces venezuelae. AB - The cytochrome P-450 PikC from Streptomyces venezuelae exhibits significant substrate tolerance and performs multiple hydroxylation reactions on structurally variant macrolides bearing the deoxyamino sugar desosamine. In previously determined co-crystal structures (Sherman, D. H., Li, S., Yermalitskaya, L. V., Kim, Y., Smith, J. A., Waterman, M. R., and Podust, L. M. (2006) J. Biol. Chem. 281, 26289-26297), the desosamine moiety of the native substrates YC-17 and narbomycin is bound in two distinct buried and surface-exposed binding pockets, mediated by specific interactions between the protonated dimethylamino group and the acidic amino acid residues Asp(50), Glu(85), and Glu(94). Although the Glu(85) and Glu(94) negative charges are essential for maximal catalytic activity of native enzyme, elimination of the surface-exposed negative charge at Asp(50) results in significantly enhanced catalytic activity. Nevertheless, the D50N substitution could not rescue catalytic activity of PikC(E94Q) based on lack of activity in the corresponding double mutant PikC(D50N/E94Q). To address the specific role for each desosamine-binding pocket, we analyzed the x-ray structures of the PikC(D50N) mutant co-crystallized with narbomycin (1.85A resolution) and YC-17 (3.2A resolution). In PikC(D50N), the desosamine moiety of both YC-17 and narbomycin was bound in a catalytically productive "buried site." This finding suggested a two-step substrate binding mechanism, whereby desosamine is recognized in the two subsites to allow the macrolide substrate to sequentially progress toward a catalytically favorable orientation. Collectively, the binding, mutagenesis, kinetic, and x-ray structural data suggest that enhancement of the catalytic activity of PikC(D50N) is due to the facilitated relocation of substrate to the buried site, which has higher binding affinity, as opposed to dissociation in solution from the transient "surface-exposed site." PMID- 19124460 TI - Accumulation of Pax2 transactivation domain interaction protein (PTIP) at sites of DNA breaks via RNF8-dependent pathway is required for cell survival after DNA damage. AB - Genomic stability in eukaryotic cells is maintained by the coordination of multiple cellular events including cell cycle checkpoint, DNA repair, transcription, and apoptosis after DNA damage. Pax2 transactivation domain interaction protein (PTIP), a protein that contains six BRCT domains, has been implicated in DNA damage response. In this study we showed that recruitment of PTIP to damaged chromatin depends on DNA damage signaling proteins gammaH2AX.MDC1.RNF8, which in turn facilitates sustained localization of PA1 (PTIP-associated protein 1) to sites of DNA break. Similar to PTIP, depletion of PA1 increases cellular sensitivity to ionizing radiation. Furthermore, we demonstrated that the N-terminal PA1 binding domain and the C-terminal focus localization domain of PTIP are critical for PTIP function in DNA damage repair. Interestingly, although PTIP and PA1 associate with MLL (mixed lineage leukemia) complexes and participate in transcriptional regulation, this function of PTIP.PA1 in DNA damage response is likely to be independent of the MLL complexes. Taken together, we propose that a subset of PTIP.PA1 complex is recruited to DNA damage sites via the RNF8-dependent pathway and is required for cell survival in response to DNA damage. PMID- 19124461 TI - Zinc finger transcription factor INSM1 interrupts cyclin D1 and CDK4 binding and induces cell cycle arrest. AB - INSM1 is a zinc finger transcription factor that plays an important role in pancreatic beta-cell development. To further evaluate its role in cell fate determination, we investigated INSM1 effects on cell cycle function. The cyclin box of cyclin D1 is essential for INSM1 binding. Competitive pull-down and co immunoprecipitation revealed that INSM1 binding to cyclin D1 interrupts its association with CDK4 and induces hypophosphorylation of the retinoblastoma protein. An inducible Tet-on system was established in Cos-7 and Panc-1 cells. Using serum starvation, we synchronized the cell cycle and subsequently induced cell cycle progression by serum stimulation. Comparison of the INSM1 induction group with the noninduced control group, INSM1 ectopic expression causes cell cycle arrest, whereas the INSM1-mediated cell cycle arrest could be reversed by cyclin D1 and CDK4 overexpression. The proline-rich N-terminal portion of INSM1 is required for cyclin D1 binding. Mutation of proline residues abolished cyclin D1 binding and also diminished its ability to induce cell cycle arrest. Cellular proliferation of Panc-1 cells was inhibited by INSM1 overexpression demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, soft agar colony formation, as well as tumor growth in a nude mouse model. Taken together, we provide evidence to support that INSM1 binds to cyclin D1, interrupts cell cycle signaling, and inhibits cellular proliferation. PMID- 19124462 TI - The inhibitory mechanism of protein synthesis by YoeB, an Escherichia coli toxin. AB - YoeB is a toxin encoded by the yefM-yoeB antitoxin-toxin operon in the Escherichia coli genome. Here we show that YoeB, a highly potent protein synthesis inhibitor, specifically blocks translation initiation. In in vivo primer extension experiments using two different mRNAs, a major band was detected after YoeB induction at three bases downstream of the initiation codon at 2.5 min. An identical band was also detected in in vitro toeprinting experiments after the addition of YoeB to the reaction mixtures containing 70 S ribosomes and the same mRNAs, even in the absence of tRNA(f)(Met). Notably, this band was not detected in the presence of YoeB alone, indicating that YoeB by itself does not have endoribonuclease activity under the conditions used. The 70 S ribosomes increased upon YoeB induction, and YoeB was found to be specifically associated with 50 S subunits. Using tetracycline and hygromycin B, we demonstrated that YoeB binds to the 50 S ribosomal subunit in 70 S ribosomes and interacts with the A site leading to mRNA cleavage at this site. As a result, the 3'-end portion of the mRNA was released from ribosomes, and translation initiation was effectively inhibited. These results demonstrate that YoeB primarily inhibits translation initiation. PMID- 19124463 TI - Control of liver glycogen synthase activity and intracellular distribution by phosphorylation. AB - Eukaryotic glycogen synthase activity is regulated by reversible phosphorylation at multiple sites. Of the two GS isoforms found in mammals, the muscle enzyme (muscle glycogen synthase) has received more attention and the relative importance of every known phosphorylation site in the control of its activity and intracellular distribution has been previously addressed. We have analyzed the impact of the dephosphorylation at the homologous sites of the glycogen synthase liver (LGS) isoform. Serine residues at these sites were replaced by non phosphorylatable alanine residues, singly or in pairs, and the resultant LGS variants were expressed in cultured cells using adenoviral vectors. The sole mutation at site 2 (Ser7) yielded an enzyme that was almost fully active and able to induce glycogen deposition in primary hepatocytes incubated in the absence of glucose and in FTO2B cells, a cell line that does not normally synthesize glycogen. Mutation at site 2 was also sufficient to trigger the aggregation and translocation of LGS from the cytoplasm to the hepatocyte cell cortex in the absence of glucose. However, this redistribution was not observed in hepatocytes incubated without glucose when an additional mutation (E509A), which renders the enzyme inactive, was introduced. This result suggests that LGS translocation is strictly dependent on glycogen synthesis. PMID- 19124464 TI - Tyrosine kinase activity of epidermal growth factor receptor is regulated by GM3 binding through carbohydrate to carbohydrate interactions. AB - Epidermal growth factor receptor (EGFR), an N-glycosylated transmembrane protein with an intracellular kinase domain, undergoes dimerization by ligand binding resulting in activation of the kinase domain and phosphorylation. Ganglioside GM3 containing sialyllactose inhibits the tyrosine kinase activity of EGFR through carbohydrate to carbohydrate interactions (CCI) between N-glycans with GlcNAc termini on EGFR and oligosaccharides on GM3. In this study, we provide further evidence for CCI between EGFR and GM3. (i) In vitro and in situ, the inhibitory effect of GM3 on EGFR tyrosine kinase was much higher in A431 cells upon exposure of the GlcNAc termini of the N-glycans to glycosidase treatment (neuraminidase and beta-galactosidase) than in untreated A431 cells. Furthermore, the GM3 mediated inhibition was abrogated by co-incubation with N-glycan containing terminal GlcNAc. (ii) In situ, inhibition of EGFR phosphorylation by GM3 was not observed in alpha-mannosidase IB (ManIB)-knocked down A431 cells that accumulate high mannose-type N-glycans. (iii) EGFR binding to GM3 was enhanced in glycosidase-treated cells that accumulated GlcNAc termini, whereas GM3 did not bind to EGFR from ManIB-knocked down cells that accumulated high mannose-type N glycans. These results indicate that GM3-mediated inhibition of EGFR phosphorylation is caused by interaction of GM3 with GlcNAc-terminated N-glycan on EGFR. PMID- 19124465 TI - Mechanistic studies of the bypass of a bulky single-base lesion catalyzed by a Y family DNA polymerase. AB - 1-nitropyrene, the most abundant nitro polycyclic aromatic hydrocarbon in diesel emissions, has been found to react with DNA to form predominantly N (deoxyguanosin-8-yl)-1-aminopyrene (dGAP). This bulky adduct has been shown to induce genetic mutations, which may implicate Y-family DNA polymerases in its bypass in vivo. To establish a kinetic mechanism for the bypass of such a prototype single-base lesion, we employed pre-steady-state kinetic methods to investigate individual nucleotide incorporations upstream, opposite, and downstream from a site-specifically placed dGAP lesion catalyzed by Sulfolobus solfataricus DNA polymerase IV (Dpo4), a model Y-family DNA polymerase. Dpo4 was able to bypass dGAP but paused strongly at two sites: opposite the lesion and immediately downstream from the lesion. Both nucleotide incorporation efficiency and fidelity decreased significantly at the pause sites, especially during extension of the bypass product. Interestingly, a 4-fold tighter binding affinity of damaged DNA to Dpo4 promoted catalysis through putative interactions between the active site residues of Dpo4 and 1-aminopyrene moiety at the first pause site. In the presence of a DNA trap, the kinetics of nucleotide incorporation at these sites was biphasic in which a small, fast phase preceded a larger, slow phase. In contrast, only a large, fast phase was observed during nucleotide incorporation at non-pause sites. Our kinetic studies support a general kinetic mechanism for lesion bypass catalyzed by numerous DNA polymerases. PMID- 19124466 TI - Regulation of autophagy by the p300 acetyltransferase. AB - Autophagy is a regulated process of intracellular catabolism required for normal cellular maintenance, as well as serving as an adaptive response under various stress conditions, including starvation. The molecular regulation of autophagy in mammalian cells remains incompletely understood. Here we demonstrate a role for protein acetylation in the execution and regulation of autophagy. In particular, we demonstrate that the p300 acetyltransferase can regulate the acetylation of various known components of the autophagy machinery. Knockdown of p300 reduces acetylation of Atg5, Atg7, Atg8, and Atg12, although overexpressed p300 increases the acetylation of these same proteins. Furthermore, p300 and Atg7 colocalize within cells, and the two proteins physically interact. The interaction between p300 and Atg7 is dependent on nutrient availability. Finally, we demonstrate that knockdown of p300 can stimulate autophagy, whereas overexpression of p300 inhibits starvation-induced autophagy. These results demonstrate a role for protein acetylation and particularly p300 in the regulation of autophagy under conditions of limited nutrient availability. PMID- 19124467 TI - ARF6 regulates the synthesis of fusogenic lipids for calcium-regulated exocytosis in neuroendocrine cells. AB - An important role for specific lipids in membrane fusion has recently emerged, but regulation of their biosynthesis remains poorly understood. Among fusogenic lipids, phosphatidic acid and phosphoinositol 4,5-bisphosphate (PIP(2)) have been proposed to act at various steps of neurotransmitter and hormone exocytosis. Using real time FRET (fluorescence resonance energy transfer) measurements, we show here that the GTPase ARF6, potentially involved in the synthesis of these lipids, is activated at the exocytotic sites in PC12 cells stimulated for secretion. Depletion of endogenous ARF6 by siRNA dramatically inhibited secretagogue-evoked exocytosis. ARF6-siRNA greatly reduced secretagogue-evoked phospholipase D (PLD) activation and phosphatidic acid formation at the plasma membrane and moderately reduced constitutive levels of PIP(2) present at the plasma membrane in resting cells. Expression of an ARF6 insensitive to short interference RNA (siRNA) fully rescued secretion in ARF6-depleted cells. However, a mutated ARF6 protein specifically impaired in its ability to stimulate PLD had no effect. Finally, we show that the ARF6-siRNA-mediated inhibition of exocytosis could be rescued by an exogenous addition of lysophosphatidylcholine, a lipid that favors negative curvature on the inner leaflet of the plasma membrane. Altogether these data indicate that ARF6 is a critical upstream signaling element in the activation of PLD necessary to produce the fusogenic lipids required for exocytosis. PMID- 19124468 TI - Formation of a stabilized cysteine sulfinic acid is critical for the mitochondrial function of the parkinsonism protein DJ-1. AB - The formation of cysteine-sulfinic acid has recently become appreciated as a modification that links protein function to cellular oxidative status. Human DJ 1, a protein associated with inherited parkinsonism, readily forms cysteine sulfinic acid at a conserved cysteine residue (Cys106 in human DJ-1). Mutation of Cys106 causes the protein to lose its normal protective function in cell culture and model organisms. However, it is unknown whether the loss of DJ-1 protective function in these mutants is due to the absence of Cys106 oxidation or the absence of the cysteine residue itself. To address this question, we designed a series of substitutions at a proximal glutamic acid residue (Glu18) in human DJ-1 that alter the oxidative propensity of Cys106 through changes in hydrogen bonding. We show that two mutations, E18N and E18Q, allow Cys106 to be oxidized to Cys106-sulfinic acid under mild conditions. In contrast, the E18D mutation stabilizes a cysteine-sulfenic acid that is readily reduced to the thiol in solution and in vivo. We show that E18N and E18Q can both partially substitute for wild-type DJ-1 using mitochondrial fission and cell viability assays. In contrast, the oxidatively impaired E18D mutant behaves as an inactive C106A mutant and fails to protect cells. We therefore conclude that formation of Cys106 sulfinic acid is a key modification that regulates the protective function of DJ 1. PMID- 19124469 TI - Glucocorticoid receptor activation of the Ciz1-Lcn2 locus by long range interactions. AB - The cellular response to glucocorticoid receptor (GR) activation involves a highly orchestrated series of regulatory actions influenced at multiple levels by a variety of mechanisms including the action of transcription factors and chromatin modifiers. Because the majority of GR binding sites (glucocorticoid responsive elements (GREs)) are distant from promoters, it is likely that interactions at a distance play an important role in GR action. To determine whether long range chromosomal associations play a role in transcription regulation by GR, we utilized a chromosome conformation capture-based technique (associated chromosome trap) to identify unknown, remote sequences that interact with the GR-induced Lipocalin2 (Lcn2) gene. Our screen revealed that the Lcn2 GRE interacts with the Ciz1 gene, nearly 30 kb upstream. Ciz1 was subsequently found to be a novel GR-responsive gene. The GRE proximal to the Lcn2 promoter apparently functions to regulate both the Lcn2 gene and the distal Ciz1 gene. Using quantitative chromosome conformation capture, we find that a loop structure is organized between these two genes. This structure is hormone-independent and present only in cell types where the genes are active. The strong correlation between gene expression and loop structure in different cell lines suggests that high order interactions play a role in determining tissue-specific gene regulation. PMID- 19124471 TI - Enhancing the receptor affinity of the sialic acid-binding domain of Vibrio cholerae sialidase through multivalency. AB - Many glycoside hydrolases possess carbohydrate-binding modules (CBMs) that help target these enzymes to appropriate substrates and increase their catalytic efficiency. The Vibrio cholerae sialidase contains two CBMs, one of which is designated as a family CBM40 module and has been shown through structural and calorimetry studies to recognize the alpha-anomer of sialic acid with a KD of approximately 30 microM at 37 degrees C. The affinity of this V. cholerae CBM40 module for sialic acid is one of the highest reported for recognition of a monosaccharide by a CBM. As Nature often increases a weak substrate affinity through multivalency, we have explored the potential of developing reagents with an increased affinity for sialic acid receptors through linking CBM40 modules together. The V. cholerae CBM40 was subcloned and crystallized in the presence of sialyllactose confirming its ability to recognize sialic acid. Calorimetry revealed that this CBM40 demonstrated specificity to alpha(2,3)-, alpha(2,6)-, and alpha(2,8)-linked sialosides. Polypeptides containing up to four CBM40 modules in tandem were created to determine if an increase in affinity to sialic acid could be achieved through an avidity effect. Using SPR and a multivalent alpha(2,3)-sialyllactose ligand, we show that increasing the number of linked modules does increase the affinity for sialic acid. The four-CBM40 module protein has a 700- to 1500-fold increase in affinity compared with the single-CBM40 module. Varying the linker length of amino acids between each CBM40 module had little effect on the binding of these polypeptides. Finally, fluorescence activated cell sorting analysis demonstrated that a green fluorescent protein fused to three CBM40 modules bound to subpopulations of human leukocytes. These studies lay the foundation for creating high affinity, multivalent CBMs that could have broad application in glycobiology. PMID- 19124470 TI - The crystal structure of galacto-N-biose/lacto-N-biose I phosphorylase: a large deformation of a TIM barrel scaffold. AB - Galacto-N-biose/lacto-N-biose I phosphorylase (GLNBP) from Bifidobacterium longum, a key enzyme for intestinal growth, phosphorolyses galacto-N-biose and lacto-N-biose I with anomeric inversion. GLNBP homologues are often found in human pathogenic and commensal bacteria, and their substrate specificities potentially define the nutritional acquisition ability of these microbes in their habitat. We report the crystal structures of GLNBP in five different ligand binding forms. This is the first three-dimensional structure of glycoside hydrolase (GH) family 112. The GlcNAc- and GalNAc-bound forms provide structural insights into distinct substrate preferences of GLNBP and its homologues from pathogens. The catalytic domain consists of a partially broken TIM barrel fold that is structurally similar to a thermophilic beta-galactosidase, strongly supporting the current classification of GLNBP homologues as one of the GH families. Anion binding induces a large conformational change by rotating a half unit of the barrel. This is an unusual example of molecular adaptation of a TIM barrel scaffold to substrates. PMID- 19124472 TI - Redox- and calmodulin-dependent S-nitrosylation of the KCNQ1 channel. AB - Nitric oxide (NO) is a gaseous signal mediator showing numerous important biological effects. NO has been shown in many instances to exhibit its action via the protein S-nitrosylation mechanism, in which binding of NO to Cys residues regulate protein function independently of activation of soluble guanylate cyclase. The direct link between protein S-nitrosylation and functional modulation, however, has been demonstrated only in limited examples. Furthermore, although most proteins have more than one Cys residue, the mechanism by which a certain Cys becomes a specific target residue of S-nitrosylation is poorly understood. We have previously reported that NO regulates currents through the cardiac slowly activating delayed rectifier potassium channel (I(Ks)) irrespective of soluble guanylate cyclase activation. Here we demonstrate using a biotin-switch assay that NO induced S-nitrosylation of the alpha-subunit of the I(Ks) channel, KCNQ1, at Cys(445) in the C terminus. A redox motif flanking Cys(445) and the interaction of KCNQ1 with calmodulin are required for preferential S-nitrosylation of Cys(445). A patch clamp experiment shows that S nitrosylation of Cys(445) modulates the KCNQ1/KCNE1 channel function. Our data provide a molecular basis of NO-mediated regulation of the I(Ks) channel. This novel regulatory mechanism of the I(Ks) channel may play a role in previously demonstrated NO-mediated phenomenon in cardiac electrophysiology, including shortening in action potential duration in response to intracellular Ca(2+) or sex hormones. PMID- 19124473 TI - Functional cross-interaction of the fragments produced by the cleavage of distinct adhesion G-protein-coupled receptors. AB - The unusual adhesion G-protein-coupled receptors (aGPCRs) contain large extracellular N-terminal domains, which resemble cell-adhesion receptors, and C terminal heptahelical domains, which may couple to G-proteins. These receptors are cleaved post-translationally between these domains into two fragments (NTF and CTF). Using the aGPCR latrophilin 1, we previously demonstrated that the fragments behave as independent cell-surface proteins. Upon binding the agonist, alpha-latrotoxin (LTX), latrophilin fragments reassemble and induce intracellular signaling. Our observations raised important questions: is the aGPCR signaling mediated by reassembled fragments or by any non-cleaved receptors? Also, can the fragments originating from distinct aGPCRs form hybrid complexes? To answer these questions, we created two types of chimerical constructs. One contained the CTF of latrophilin joined to the NTF of another aGPCR, EMR2; the resulting protein did not bind LTX but, similar to latrophilin, could couple to G-proteins. In another construct, the NTF of latrophilin was fused with the C terminus of neurexin; this chimera bound LTX but could not signal via G-proteins. Both constructs were efficiently cleaved in cells. When the two constructs were co expressed, their fragments could cross-interact, as shown by immunoprecipitation. Furthermore, LTX(N4C) induced intracellular Ca2+ signaling only in cells expressing both constructs but not each individual construct. Finally, we demonstrated that fragments of unrelated aGPCRs can be cross-immunoprecipitated from live tissues. Thus, (i) aGPCR fragments behave as independent proteins, (ii) the complementary fragments from distinct aGPCRs can cross-interact, and (iii) these cross-complexes are functionally active. This unusual cross-assembly of aGPCR fragments could couple cell-surface interactions to multiple signaling pathways. PMID- 19124474 TI - Microarrays and epidemiology: ensuring the impact and accessibility of research findings. PMID- 19124475 TI - NewGeneris: a European study on maternal diet during pregnancy and child health. PMID- 19124476 TI - Physical activity and postmenopausal breast cancer: proposed biologic mechanisms and areas for future research. AB - Convincing evidence now supports a probable preventive role for physical activity in postmenopausal breast cancer. The mechanisms by which long-term physical activity affect risk, however, remain unclear. The aims of this review were to propose a biological model whereby long-term physical activity lowers postmenopausal breast cancer risk and to highlight gaps in the epidemiologic literature. To address the second aim, we summarized epidemiologic literature on 10 proposed biomarkers, namely, body mass index (BMI), estrogens, androgens, sex hormone binding globulin, leptin, adiponectin, markers of insulin resistance, tumor necrosis factor-alpha, interleukin-6, and C-reactive protein, in relation to postmenopausal breast cancer risk and physical activity, respectively. Associations were deemed "convincing," "probable," "possible," or "hypothesized" using set criteria. Our proposed biological model illustrated the co-occurrence of overweight/obesity, insulin resistance, and chronic inflammation influencing cancer risk through interrelated mechanisms. The most convincing epidemiologic evidence supported associations between postmenopausal breast cancer risk and BMI, estrogens, and androgens, respectively. In relation to physical activity, associations were most convincing for BMI, estrone, insulin resistance, and C reactive protein. Only BMI and estrone were convincingly (or probably) associated with both postmenopausal breast cancer risk and physical activity. There is a need for prospective cohort studies relating the proposed biomarkers to cancer risk and for long-term exercise randomized controlled trials comparing biomarker changes over time, specifically in postmenopausal women. Future etiologic studies should consider interactions among biomarkers, whereas exercise trials should explore exercise effects independently of weight loss, different exercise prescriptions, and effects on central adiposity. PMID- 19124477 TI - Sampling strategies for tissue microarrays to evaluate biomarkers in ovarian cancer. AB - INTRODUCTION: Tissue microarrays (TMA) enable rapid analysis of biomarkers in large-scale studies involving archival tumor specimens, however, their utility in heterogeneous tumors such as ovarian cancer is limited. METHODS: In this study, immunohistochemical analysis was done on TMAs comprised of epithelial ovarian cancer (EOC) to estimate the prevalence of loss of expression of three mismatch repair proteins. TMAs were initially created using cores sampled from the center of donor tissue blocks from 59 EOC cases. Full sections were subsequently created and levels of expression were compared between tissues sampled from the central portion versus the periphery. Follow-up analyses were done by obtaining cores from the periphery of up to five additional donor blocks per case. A linear mixed model for each protein was used to investigate differences between results from the initial and follow-up blocks. RESULTS: In the original TMAs created using centrally sampled cores, loss of mismatch repair expression was noted in 17 (29%) of the 59 cases. By comparison, analyses from peripherally sampled cores revealed loss of expression in only 6 of these 17 cases. For each protein, significant differences (P < 0.05) were detected between results from the initial donor block and the majority of the follow-up blocks. CONCLUSIONS: Our investigations, based on EOC, suggest that sampling variability in protein expression may result when TMAs are used. Thus, at least for EOC, it is important to preferentially sample from the periphery of tumor blocks where exposure to tissue fixatives is optimal. PMID- 19124478 TI - Effects of menthol on tobacco smoke exposure, nicotine dependence, and NNAL glucuronidation. AB - Menthol is a controversial cigarette additive because its physiologic or pharmacologic effects may possibly increase the risk for cancer and its targeted market is the Black community. In a community-based cross-sectional study on 525 Black and White volunteers, we compared levels of urinary and plasma cotinine, plasma thiocyanate, urinary 4-(N-nitrosomethylamino)-1-(3-pyridyl)-1-butanol (NNAL), and its detoxified form (NNAL-Gluc) between menthol and nonmenthol smokers. In regression models that adjusted for daily cigarette intake, no significant differences were observed in the concentration of these biomarkers by menthol status in both races. There was no significant association between high Fagerstrom nicotine dependence scores and the use of menthol cigarettes (odds ratio, 1.1; 95% confidence interval, 0.6-2.0), but an increased risk was observed with smoking a cigarette soon (or=55 years, assumed too old to be in perimenopause. The main end point was breast cancer death. Breast cancer survival was estimated by the Kaplan-Meier method. The hazard ratios of breast cancer death by retinol level were estimated by Cox models stratified for age, where relevant, and recruitment period, and adjusted for tumor size and histology. RESULTS: At 12 years, patients with low retinol (<2.08 micromol/L, median of distribution) had lower breast cancer survival than those with high retinol (log rank P = 0.052); the difference was significant for women >or=55 years (log-rank P = 0.006). The adjusted hazard ratios for low versus high retinol were 2.11 (95% confidence interval, 1.08-4.14) for all women and 3.58 (95% confidence interval, 1.50-8.57) for those >or=55 years. CONCLUSIONS: Low plasma retinol strongly predicts poorer prognosis in postmenopausal breast cancer patients. Retinol levels should be determined as part of the prognostic workup. PMID- 19124480 TI - Serum undercarboxylated osteocalcin as biomarker of vitamin K intake and risk of prostate cancer: a nested case-control study in the Heidelberg cohort of the European prospective investigation into cancer and nutrition. AB - From cell studies, Vitamin K is known to exert anticancer effects on a variety of cancer cell lines, including prostate cancer cells. Recently, we reported an inverse association between dietary intake of menaquinones (vitamin K(2)), but not phylloquinone (vitamin K(1)), and risk of prostate cancer. In this nested case-control study including 250 prostate cancer cases and 494 matched controls, we aimed to confirm this cancer-protective effect using serum undercarboxylated osteocalcin (ucOC), a biomarker of vitamin K status inversely associated with vitamin K intake. In addition, effect modification by a functionally relevant polymorphism in the vitamin K epoxide reductase gene (VKORC1) was assessed. Serum ucOC and intact total osteocalcin (iOC) were analyzed with the use of ELISA tests. Serum ucOC was expressed relative to iOC (i.e., as ucOC/iOC ratio). Conditional logistic regression was used to calculate multivariate adjusted odds ratios (OR) and 95% confidence intervals (95% CI). Serum ucOC/iOC ratio was positively associated with advanced-stage (OR per 0.1 increment, 1.38; 95% CI, 1.03-1.86) and high-grade prostate cancer (OR, 1.21; 95% CI, 1.00-1.46) but not with total prostate cancer. The significant association with advanced-stage prostate cancer was confirmed when serum ucOC/iOC ratio was jointly modeled with menaquinone intake data. There was indication of a lower prostate cancer risk in carriers of the A allele (compared with GG carriers) of the +2255 VKORC1 polymorphism with increasing menaquinone intake (P(interaction) = 0.14) whereas no distinct effect modification was observed for the ucOC/iOC ratio (P(interaction) = 0.37). The increased risks of advanced-stage and high-grade prostate cancer with higher serum ucOC/iOC ratio strengthen the findings for dietary menaquinone intake. PMID- 19124481 TI - The elevated expression of a mismatch repair protein is a predictor for biochemical recurrence after radical prostatectomy. AB - PURPOSE: The inability to predict clinical outcome of prostate cancer is a major impediment to effective treatment decisions and patient counseling. New markers of recurrence are needed to improve the accuracy of risk assessment and treatment of prostate cancer. Our previous studies identified a mismatch repair protein, PMS2, to be elevated in prostate cancer; here, we investigate the prognostic potential of this marker. We hypothesized that the elevation of PMS2 would correlate with disease outcome. EXPERIMENTAL DESIGN: Retrospective quantitative immunohistochemistry was done to measure PMS2 in high-grade cancers of 166 men treated by radical prostatectomy with a biochemical recurrence rate of 56%. Associations between PMS2 levels, pathologic variables, and biochemical recurrence over time were determined. RESULTS: The mean level of PMS2 protein was consistently higher in both cancer-associated benign epithelium and cancer cells of patients who recurred, compared with nonrecurrent patients. PMS2 was an independent predictor of time-to-recurrence in Cox multivariate analyses and significantly stratified patients based on outcome. PMS2 was able to improve the sensitivity of total percent Gleason 4/5 as a risk factor for recurrence in this cohort. CONCLUSIONS: PMS2 protein levels were shown to be a predictor of time-to recurrence after surgery. This study is the first to document that the elevation of a mismatch repair protein negatively correlates with prognosis and has implications in patient diagnosis and molecular profiling. PMID- 19124482 TI - TGF-alpha expression as a potential biomarker of risk within the normal-appearing colorectal mucosa of patients with and without incident sporadic adenoma. AB - BACKGROUND: Transforming growth factor-alpha (TGF-alpha), a stimulatory growth factor and member of the epidermal growth factor family, is a mediator of oncogenesis and malignant progression in colorectal carcinogenesis. Limited evidence suggests its utility as a growth-related biomarker of risk for colorectal cancer. METHODS: We measured expression of TGF-alpha in biopsies of normal-appearing colorectal mucosa using automated immunohistochemistry and quantitative image analysis in a subsample of 29 cases and 31 controls from a colonoscopy-based case-control study (n = 203) of biomarkers of risk for incident sporadic colorectal adenoma. Diet, lifestyle, and medical history were assessed with validated questionnaires. RESULTS: TGF-alpha expression in the rectum was 51% higher in cases compared with controls (P = 0.05) and statistically significantly associated with accepted risk factors for colorectal neoplasms (36% lower among nonsteroidal anti-inflammatory drug users, 49% lower among women using hormone replacement therapy, 79% higher among persons with a family history of colorectal cancer). CONCLUSIONS: TGF-alpha expression in the normal-appearing rectal mucosa shows promise as an early, potentially modifiable biomarker of risk for colorectal cancer. PMID- 19124483 TI - Urinary melatonin levels and postmenopausal breast cancer risk in the Nurses' Health Study cohort. AB - BACKGROUND: Melatonin seems to play a role in breast cancer etiology, but data addressing the association between melatonin levels and breast cancer risk in postmenopausal women is sparse. METHODS: We conducted a nested case-control study in the Nurses' Health Study cohort. First spot morning urine was collected from 18,643 cancer-free women from March 2000 through December 2002. The concentration of the major metabolite of melatonin, 6-sulfatoxymelatonin (aMT6s), was available for 357 postmenopausal women who developed incident breast cancer through May 31, 2006, along with 533 matched control subjects. We used multivariable conditional logistic regression models to investigate associations. All statistical tests were two sided. RESULTS: An increased concentration of urinary aMT6s was statistically significantly associated with a lower risk of breast cancer (odds ratio for the highest versus lowest quartile of morning urinary aMT6s, 0.62; 95% confidence interval, 0.41-0.95; P(trend) = 0.004). There was no apparent modification of risk by hormone receptor status of breast tumors, age, body mass index, or smoking status. CONCLUSION: Results from this prospective study add substantially to the growing literature that supports an inverse association between melatonin levels and breast cancer risk. PMID- 19124484 TI - Coexpression of alpha6beta4 integrin and guanine nucleotide exchange factor Net1 identifies node-positive breast cancer patients at high risk for distant metastasis. AB - Preclinical data indicate that alpha6beta4 integrin signaling through Ras homolog gene family, member A, plays an important role in tumor cell motility. The objective of this study was to determine whether the combined expression of alpha6beta4 integrin and neuroepithelioma transforming gene 1 (Net1), a guanine nucleotide exchange factor specific for Ras homolog gene family member A, is associated with adverse clinical outcome in breast cancer patients. Immunohistochemical expression of each protein was evaluated in a tumor tissue microarray prepared from the primary tumors of 94 node-positive patients with invasive breast carcinoma treated with total mastectomy and doxorubicin-based chemotherapy without radiation with a median follow-up of 12.5 years. Associations between staining results and multiple clinicopathologic variables were investigated. Although there was no significant association between alpha6beta4 integrin or Net1 expression and clinical outcome when each marker was considered individually, coexpression of alpha6beta4 and Net1 was associated with decreased distant metastasis-free survival (P = 0.030). In the subset of patients with hormone receptor-positive tumors, coexpression of alpha6beta4 and Net1 was associated with a decrease in distant metastasis-free and overall survival (P < 0.001 and P = 0.006, respectively). Although an association between human epidermal growth factor receptor 2 expression and coexpression of alpha6beta4 and Net1 (P = 0.008) was observed, coexpression of alpha6beta4 and Net1 (hazard ratio, 1.63; P = 0.02) and lymphovascular invasion (hazard ratio, 2.35; P = 0.02) were the only factors independently associated with the development of distant metastasis in multivariate analysis. These findings suggest that coexpression of alpha6beta4 integrin and Net1 could be a useful biomarker for aggressive disease in node-positive breast cancer patients. PMID- 19124486 TI - Seasonal and regional contributors of 1-hydroxypyrene among children near a steel mill. AB - Urinary 1-hydroxypyrene (1-OHP) is a biomarker of exposure to polycyclic aromatic hydrocarbons (PAH). Effect of residence on children's PAH exposure was reported among children living near a polluted area. Instead of a snapshot assessment, however, a temporal history of exposure characteristics needs to be assessed in the studies of chronic disease development such as cancer. The urinary 1-OHP measurements were repeated to determine regional effect of ambient air pollution on 1-OHP levels over extended periods. Two sites were chosen: (a) one site located near the steel mill ("nearby" site) and (b) the other site located at a longer distance from the mill ("remote" site). Spot urinary 1-OHP levels were measured from 72 children for 3 consecutive days per month, repeated over 9-month period. Compared with remote site, the nearby site had increased the urinary 1 OHP level by 62.3% [95% confidence interval (95% CI), 39.8-88.3%]. Other statistically significant factors that contributed to the level include sex [16.5% (95% CI, 1.2-34.1%) higher for girls than boys], consumption of charbroiled meat [16.5% (95% CI, 1.1-34.2%) higher], and an increase in PM(10) [10.1% (95% CI, 4.8-15.7%) higher for the interquartile range increment]. Controlling for covariates, the 1-OHP levels were increased in the summer and fall compared with winter. The magnitude of the effects of both seasons had diminished after adjusting for PM(10). This is the first report providing seasonal and regional contributors to environmental PAH exposure, assessed by urinary 1-OHP, with higher 1-OHP levels during summer when ambient pollution was also high. PMID- 19124485 TI - Physical activity and risk of recurrence and mortality in breast cancer survivors: findings from the LACE study. AB - INTRODUCTION: Identifying modifiable factors that reduce the risk of recurrence and improve survival in breast cancer survivors is a pressing concern. The purpose of this study was to examine the association of physical activity following diagnosis and treatment with the risk of breast cancer recurrence and mortality and all-cause mortality in women with early-stage breast cancer. MATERIALS AND METHODS: The sample consisted of 1,970 women from the Life After Cancer Epidemiology study, a prospective investigation of behavioral risk factors and health outcomes. Self-reported frequency and duration of work-related, household and caregiving, recreational, and transportation-related activities during the six months prior to enrollment were assessed. Outcomes were ascertained from electronic or paper medical charts. Hazard ratios and 95% confidence intervals were estimated from delayed entry Cox proportional hazards models. RESULTS: Although age-adjusted results suggested that higher levels of physical activity were associated with reduced risk of recurrence and breast cancer mortality (P for trend = 0.05 and 0.07, respectively for highest versus lowest level of hours per week of moderate physical activity), these associations were attenuated after adjustment for prognostic factors and other confounding variables (P for trend = 0.36 and 0.26). In contrast, a statistically significant protective association between physical activity and all-cause mortality remained in multivariable analyses (hazard ratio, 0.66; 95% confidence interval, 0.42 1.03; P for trend = 0.04). CONCLUSIONS: These findings do not support a protective effect of physical activity on breast cancer recurrence or mortality but do suggest that regular physical activity is beneficial for breast cancer survivors in terms of total mortality. PMID- 19124487 TI - The relative importance of genetics and environment on mammographic density. AB - BACKGROUND: Although several environmental factors predict mammographic density, estimates of its heritability have been quite high. We investigated whether part of the presumed heritability might be attributed to differential sharing of modifiable risk factors in monozygotic (MZ) and dizygotic (DZ) twins. METHODS: We measured percent and absolute mammographic density using mammograms from 257 MZ and 296 DZ twin pairs. The correlation of intrapair mammographic density was compared according to zygosity across strata of modifiable risk factors. Portions of variance attributable to additive genetic factors, shared environment, and individual environment were calculated using a variance component methodology in the entire set, and within twin pairs stratified by environmental trait similarity. RESULTS: Both percent density and absolute mammographic density were more highly correlated between MZ twins than DZ twins, but the correlations varied across strata. Body mass index (BMI) and parity strongly predicted differences in mammographic density within MZ twin pairs. After adjusting for covariates, 53% of the total variance in percent density and 59% of that in absolute density seemed attributable to genetic effects, but these estimates varied greatly by stratum. For twins dissimilar on BMI (difference >2.5 kg/m(2)), the additive genetic component of absolute density was estimated at only 20% (+/ 19%), and the common and individual environment at 21% (+/-14%) and 49%, respectively (P value for heterogeneity across BMI = 0.0001). CONCLUSION: Our results confirm that the genome is an important determinant of mammographic density but suggest that an unknown portion of the mammographic density effect attributed to the genome may be due to shared modifiable environmental factors. PMID- 19124489 TI - Trends in area-socioeconomic and race-ethnic disparities in breast cancer incidence, stage at diagnosis, screening, mortality, and survival among women ages 50 years and over (1987-2005). AB - BACKGROUND: Breast cancer is the most commonly diagnosed cancer and the second leading cause of cancer death among women in the United States and varies systematically by race-ethnicity and socioeconomic status. Previous research has often focused on disparities between particular groups, but few studies have summarized disparities across multiple subgroups defined by race-ethnic and socioeconomic position. METHODS: Data on breast cancer incidence, stage, mortality, and 5-year cause-specific probability of death (100 - survival) were obtained from the Surveillance, Epidemiology, and End Results program and data on mammography screening from the National Health Interview Survey from 1987 to 2005. We used four area-socioeconomic groups based on the percentage of poverty in the county of residence (<10, 10-15, 15-20, +20%) and five race-ethnic groups (White, Black, Asian, American Indian, and Hispanic). We used summary measures of disparity based on both rate differences and rate ratios. RESULTS: From 1987 to 2004, area-socioeconomic disparities declined by 20% to 30% for incidence, stage at diagnosis, and 5-year cause-specific probability of death, and by roughly 100% for mortality, whether measured on the absolute or relative scale. In contrast, relative area-socioeconomic disparities in mammography use increased by 161%. Absolute race-ethnic disparities declined across all outcomes, with the largest reduction for mammography (56% decline). Relative race-ethnic disparities for mortality and 5-year cause-specific probability of death increased by 24% and 17%, respectively. CONCLUSIONS: Our analysis suggests progress towards race ethnic and area-socioeconomic disparity goals for breast cancer, especially when measured on the absolute scale. However, greater progress is needed to address increasing relative socioeconomic disparities in mammography and race-ethnic disparities in mortality and 5-year cause-specific probability of death. PMID- 19124488 TI - Human papillomavirus cofactors by disease progression and human papillomavirus types in the study to understand cervical cancer early endpoints and determinants. AB - Human papillomavirus (HPV) cofactors for cervical cancer include smoking, multiparity, and oral contraceptive use, but their mechanisms of action are not fully understood. It is also unknown whether cofactors vary by HPV genotypes. The Study to Understand Cervical Cancer Early Endpoints and Determinants (SUCCEED) is a cross-sectional study comprising women referred to the University of Oklahoma from November 2003 to September 2007 for abnormal cervical screening results. Detailed questionnaire data and liquid cytology specimens were collected and the latter was genotyped for HPV using the LINEAR ARRAY HPV Genotyping Test. The present analysis includes women with both questionnaire and HPV data and diagnosed with or=40 years had an odds ratio of 13.9 (95% confidence interval, 2.2-90.2) for cervical cancer compared with those aged <15 years at first arrival. Women who lived with or washed the clothes of an Australian Blue Asbestos worker did not have an increased risk for any of the gynecologic or breast cancers. DISCUSSION: There is no consistent evidence of an increased risk for gynecologic and breast cancers among the women from Wittenoom. Ovarian cancers and peritoneal mesotheliomas were not misclassified in this cohort. PMID- 19124490 TI - Incidence of ovarian, peritoneal, and fallopian tube carcinomas in the United States, 1995-2004. AB - OBJECTIVE: The objective of this analysis was to describe the distribution of pelvic carcinomas in the United States by demographic, pathologic, and clinical features. METHODS: Carcinomas of the ovary (n = 112,541), peritoneum (n = 6,458), and fallopian tube (n = 3,479) were identified through 24 population-based registries in the United States during the period 1995 to 2004. Age-adjusted incidence rates were calculated per million population using counts derived from the 2000 U.S. census. RESULTS: The age-adjusted incidence rate for ovarian carcinoma (119.9 per million) was substantially higher than for peritoneal (6.78 per million) or fallopian tube (3.72 per million) carcinomas. White women had the highest rates for all three malignancies. Rates for peritoneal carcinoma were lowest among Black women (2.88 per million) and rates for fallopian tube carcinoma were lowest among Hispanic women (2.45 per million). Serous carcinomas were the most commonly diagnosed histologic type for all anatomic sites. Peritoneal carcinomas were diagnosed at later ages (mean, 67 years) and more advanced stages (85% regional/distant) compared with fallopian tube carcinomas (mean, 64 years; 62% regional/distant) and ovarian carcinomas (mean, 63 years; 76% regional/distant). Incidence for all three pelvic carcinomas was lowest in the South. Time trend analyses between 1973 and 2005 exhibited a significant decline in ovarian carcinoma incidence and rises in the rates of peritoneal and fallopian tube cancers. CONCLUSIONS: Similarities in the incidence patterns for ovarian, peritoneal, and fallopian tube carcinomas support the likelihood of a common molecular pathogenesis. PMID- 19124492 TI - The association of bone mineral density with prostate cancer risk in the Osteoporotic Fractures in Men (MrOS) Study. AB - We investigated the association of bone mineral density (BMD) measures with prostate cancer (PCa) risk in older men enrolled in the Osteoporotic Fractures in Men Study. We hypothesized that men with higher BMD, a marker of exposure to endogenous sex hormones, would have an increased incidence of PCa. The cohort included 4,597 men (89% White, 65 years or older) with no prior history of PCa. Baseline total body, total hip, and spine BMD were assessed using dual energy X ray absorptiometry. Prostate cancer was confirmed by review of medical records. Cox regression was used to assess the association of BMD quartiles with incident PCa, adjusting for age, body mass index, and other covariates. During an average follow-up of 5.2 years, 5.6% (n = 255) of men developed PCa. Total body BMD was inversely associated with incident PCa, with a significant trend for decreasing PCa risk with increasing BMD quartiles (P(trend) = 0.007). Men in the highest total body BMD quartile had a 41% reduced risk for PCa (hazard ratio, 0.59; 95% confidence interval, 0.40-0.86), compared with men in the lowest quartile. Total hip and spine BMD did not exhibit significant relationships with PCa. Associations of BMD measures differed for low-grade (Gleason sum, 2-6) versus high-grade tumors (Gleason sum, >or=7). Significant inverse relationships with high-grade disease were noted at the total body and total hip sites. However, no associations were observed with low-grade disease. Our results provide support for an inverse association between BMD and PCa risk. Possible pathophyisological mechanisms linking BMD and PCa should be elucidated. PMID- 19124493 TI - Prenatal risk factors for cutaneous malignant melanoma: follow-up of 2,594,783 Danes born from 1950 to 2002. AB - The incidence of cutaneous malignant melanoma has increased in young adults, specifically in Denmark. In this study, we examined the risk of cutaneous malignant melanoma in relation to prenatal markers of hormone levels and to family-related risk factors. The study was based on a follow-up of 2,594,783 Danes born from 1950 to 2002. Occurrence of possible hormone-related cancers among family members and indicators of abnormal prenatal hormone levels were not associated with cutaneous malignant melanoma risk, whereas family size and mother's age at first birth were significant risk factors for cutaneous malignant melanoma in offspring. PMID- 19124495 TI - Sex hormone levels, breast cancer risk, and cancer receptor status in postmenopausal women: the ORDET cohort. AB - BACKGROUND: Endogenous sex hormone levels have been associated with increased breast cancer risk in postmenopausal women in several prospective studies. However, it remains unclear to what extent serum hormone-breast cancer associations differ with receptor status. METHODS: Associations between serum sex hormone levels and breast cancer risk were assessed in a nested case-control study on postmenopausal women of the ORDET cohort. After a median follow-up of 13.5 years, 165 women developed breast cancer. Relative risks of developing breast cancer were estimated by conditional logistic regression. RESULTS: Total and free testosterone levels were directly associated with breast cancer risk [relative risk, 3.28 (95% confidence interval, 1.93-5.55) and 2.86 (95% confidence interval, 1.66-4.94), respectively, for highest versus lowest quartile]. When relations between hormone level and risk of breast cancer expressing various receptor combinations were assessed, high total testosterone was significantly associated with increased risk of estrogen receptor-positive cancers, irrespective of progesterone receptor status. High total testosterone was also associated with increased risk of both human epidermal growth factor receptor 2 (HER2)-negative (HER2(-)) and HER2(+) cancers. High estradiol tended to be associated with increased risk of HER2(-) cancer and inversely associated with HER2(+) cancer, with significant (P = 0.027) heterogeneity between HER2(+) and HER2(-) cancers. However, there were relatively few HER2(+) cases. CONCLUSIONS: This study provides further evidence that high levels of circulating testosterone increase the risk of developing breast cancer in postmenopausal women. The cancers that develop are mainly estrogen receptor positive. Although HER2(+) and HER2(-) breast cancers were both associated with high total testosterone, they showed opposing associations with estrogen. PMID- 19124494 TI - Childhood cancer among twins and higher order multiples. AB - Although several studies have found no change or a decreased risk of childhood cancer in twins, few have controlled for potential confounders such as birth weight. We examined the association of birth plurality and childhood cancer in pooled data from five U.S. states (California, Minnesota, New York, Texas, and Washington) using linked birth-cancer registry data. The data, excluding children with Down syndrome or who died before 28 days of life, included 17,672 cases diagnosed from 1980 to 2004 at ages 28 days to 14 years and 57,966 controls with all cases and controls born from 1970 to 2004. Analyses were restricted to children weighing or=ten years duration (RR, 0.74; 95% CI, 0.56-0.96). We found a reduced risk among users of estrogen plus progestin therapy (RR, 0.78; 95% CI, 0.60-1.02), with sequential regimen users (progestin <15 days per cycle) having the largest risk reduction (RR, 0.64; 95% CI, 0.43-0.95). Past users of >or=5 years ago (RR, 0.55; 95% CI, 0.32-0.98) had the largest risk reduction. In this study, estrogen plus progestin use, especially sequential regimen use, was associated with the largest overall reduction of colorectal cancer risk. PMID- 19124499 TI - Association and interactions between DNA repair gene polymorphisms and adult glioma. AB - It is generally accepted that glioma develops through accumulation of genetic alterations. We hypothesized that polymorphisms of candidate genes involved in the DNA repair pathways may contribute to susceptibility to glioma. To address this possibility, we conducted a study on 373 Caucasian glioma cases and 365 cancer-free Caucasian controls to assess associations between glioma risk and 18 functional single-nucleotide polymorphisms in DNA repair genes. We evaluated potential gene-gene and gene-environment interactions using a multianalytic strategy combining logistic regression, multifactor dimensionality reduction and classification and regression tree approaches. In the single-locus analysis, six single-nucleotide polymorphisms [ERCC1 3' untranslated region (UTR), XRCC1 R399Q, APEX1 E148D, PARP1 A762V, MGMT F84L, and LIG1 5'UTR] showed a significant association with glioma risk. In the analysis of cumulative genetic risk of multiple single-nucleotide polymorphisms, a significant gene-dosage effect was found for increased glioma risk with increasing numbers of adverse genotypes involving the aforementioned six single-nucleotide polymorphisms (P(trend) = 0.0004). Furthermore, the multifactor dimensionality reduction and classification and regression tree analyses identified MGMT F84L as the predominant risk factor for glioma and revealed strong interactions among ionizing radiation exposure, PARP1 A762V, MGMT F84L, and APEX1 E148D. Interestingly, the risk for glioma was dramatically increased in ionizing radiation exposure individuals who had the wild-type genotypes of MGMT F84L and PARP1 A762V (adjusted odds ratios, 5.95; 95% confidence intervals, 2.21-16.65). Taken together, these results suggest that polymorphisms in DNA repair genes may act individually or together to contribute to glioma risk. PMID- 19124500 TI - Serum concentrations of cytokines and lung cancer survival in African Americans and Caucasians. AB - Accumulating evidence suggests a role for inflammation in the development and progression of cancer. Our group recently identified a cytokine gene signature in lung tissue associated with lung cancer prognosis. Therefore, we hypothesized that concentrations of circulating cytokines in serum may be associated with lung cancer survival. Ten serum cytokines, namely, interleukin (IL)-1beta, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, granulocyte macrophage colony-stimulating factor, interferon (IFN)-gamma, and tumor necrosis factor-alpha, were assessed in 353 non small cell lung cancer cases from a case-control study of lung cancer in the greater Baltimore, Maryland area. Cytokines were measured using an ultrasensitive electrochemiluminescence immunoassay. IL-6 serum concentrations (>or=4.0 pg/mL) were associated with significantly poorer survival in both African Americans [hazard ratio (HR), 2.71; 95% confidence interval (CI), 1.26-5.80] and Caucasians (HR, 1.71; 95% CI, 1.22-2.40). IL-10 (HR, 2.62; 95% CI, 1.33-5.15) and IL-12 (HR, 1.98; 95% CI, 1.14-3.44) were associated with lung cancer survival only in African Americans. Some evidence for an association of tumor necrosis factor alpha levels with survival in Caucasians was observed, although these results were not significant. These hypothesis-generating findings indicate that selected serum cytokine concentrations are associated with lung cancer survival, and indicate that further research is warranted to better understand the mechanistic underpinnings of these associations. PMID- 19124501 TI - A polymorphism in the APE1 gene promoter is associated with lung cancer risk. AB - Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential enzyme in the base excision repair pathway, which is the primary mechanism for the repair of DNA damage caused by oxidation and alkylation. We hypothesized that polymorphisms of APE1 are associated with risk for lung cancer. In the hospital-based matched case control study, a total of 730 lung cancer cases and 730 cancer-free controls were genotyped for four APE1 haplotype-tagging polymorphisms (that is, -656T>G, 400A>G, 630T>C, and 1350T>G). Among them, the single-nucleotide polymorphism 656T>G located in the promoter region of APE1 was significantly associated with risk for lung cancer. We found that, compared with -656 TT homozygotes, the variant genotypes were associated with a significantly decreased risk [adjusted odds ratio, 0.51; 95% confidence interval (95% CI), 0.33-0.79 for -656 TG; adjusted odds ratio, 0.43; 95% CI, 0.25-0.76 for -656 GG, respectively]. Furthermore, we found a statistically significant reduced risk of -656T>G variants among heavy smokers (adjusted odds ratio, 0.52; 95% CI, 0.30-0.93 for 656 TG; adjusted odds ratio, 0.27; 95% CI, 0.13-0.57 for -656 GG, respectively), with a significant gene-smoking interaction (P = 0.013). A similar gene-smoking interaction in the context of APE1 haplotypes was also observed. The in vitro promoter assay revealed that the -656 G allele had a significantly higher transcriptional activity than that of the -656 T allele. Together, our results suggest that polymorphisms of the APE1 gene possibly interact with smoking and may contribute to the development of lung cancer. PMID- 19124502 TI - Family history, genetic testing, and clinical risk prediction: pooled analysis of CHEK2 1100delC in 1,828 bilateral breast cancers and 7,030 controls. AB - If breast cancers arise independently in each breast the odds ratio (OR) for bilateral breast cancer for carriers of CHEK2 1100delC should be approximately 5.5, the square of the reported OR for a first primary (OR, 2.34). In the subset of bilateral cases with one or more affected relatives, the predicted carrier OR should be approximately 9. We have tested these predictions in a pooled set of 1,828 cases with 2 primaries and 7,030 controls from 8 studies. The second primary OR for CHEK2 1100delC carriers was 6.43 (95% confidence interval, 4.33 9.56; P < 0.0001), significantly greater than the published estimate for a first primary (P < 0.001) but consistent with its square. The predicted increase in carrier OR with increasing numbers of affected relatives was seen using bilateral cases from the UK (P(trend) = 0.0003) and Finland (P(trend) = 0.37), although not using those from the Netherlands and Russia (P = 0.001 for heterogeneity between countries). Based on a standard genetic model, we predict lifetime risks for CHEK2 1100delC carrier and noncarrier daughters of bilateral breast cancer cases of 37% and 18%, respectively. Our results imply that clinical management of the daughter of a woman with bilateral breast cancer should depend on her CHEK2 1100delC carrier status. This and other moderate penetrance breast cancer susceptibility alleles, together with family history data, will thus identify increasing numbers of women at potentially very high risk. Before such predictions are accepted by clinical geneticists, however, further population based evidence is needed on the effect of CHEK2 1100delC and other moderate penetrance alleles in women with a family history of breast cancer. PMID- 19124503 TI - Genetic polymorphisms of CYP2E1 and risk of colorectal cancer: the Fukuoka Colorectal Cancer Study. AB - Cytochrome P450 2E1 (CYP2E1) is involved in the metabolic activation of a wide variety of potential carcinogens, and functional polymorphisms in the CYP2E1 gene have been investigated in relation to colorectal cancer. We examined the relation of the CYP2E1 RsaI and 96-bp insertion polymorphisms to colorectal cancer risk and the interaction between these polymorphisms and some lifestyle risk factors. Subjects were 685 incident cases of colorectal cancer and 778 community controls. Statistical adjustment was made for alcohol use, body mass index, physical activity, and other factors. The RsaI c2 allele was associated with a decreased risk of rectal cancer [adjusted odds ratio for at least one c2 allele, 0.71; 95% confidence interval (95% CI), 0.53-0.95], and an increased risk of rectal cancer was observed among individuals having one or two 96-bp insertion alleles (adjusted odds ratio, 1.40; 95% CI, 1.06-1.85). Individuals with two 96-bp insertion alleles showed a 2.28-fold increase in colon cancer risk (95% CI, 1.29 4.01). The two polymorphisms were in almost complete linkage disequilibrium (D' = 0.94). A positive association between alcohol intake and colorectal cancer was observed only in individuals without RsaI c2 allele (P(trend) = 0.03) or in those without 96-bp insertion allele (P(trend) = 0.009). Colon cancer risk was increased in relation to red meat intake only in individuals having one or two 96 bp insertion alleles (P(interaction) = 0.03). The present study suggests that variation in activity and inducibility of CYP2E1, in relation to alcohol or red meat intake, contributes to the development of colorectal cancer. PMID- 19124505 TI - Aldehyde dehydrogenase 2 and head and neck cancer: a meta-analysis implementing a Mendelian randomization approach. AB - Alcohol drinking at high doses is a risk factor for head and neck cancer, and exposure to acetaldehyde, the principle metabolite of alcohol, is supposed to account for the increased risk. Individuals homozygous for the 2 variant allele of aldehyde dehydrogenase 2 (ALDH2) are unable to metabolize acetaldehyde, which prevents them from alcohol drinking, whereas 1 2 have 6-fold higher blood acetaldehyde concentration postalcohol consumption with respect to 1 1. According to the concept of Mendelian randomization, because this polymorphism is distributed randomly during gamete formation, its association with head and neck cancer should be not confounded by smoking. We carried out a meta-analysis of ALDH2 and head and neck cancer searching for relevant studies on Medline and Embase up to January 31, 2008, and investigated the consistency between the expected odds ratio (OR) among drinkers from the largest pooled analysis among never smokers and the observed OR from this meta-analysis by an interaction test. Six studies were selected (945 cases, 2,917 controls). The OR of head and neck cancer among 2 2 was 0.53 [95% confidence interval (95% CI), 0.28-1.00] relative to 1 1 and 1.83 (95% CI, 1.21-2.77) among 1 2. The expected OR for head and neck cancer due to alcohol intake among 1 1 was 1.38 (95% CI, 0.88-2.17) and the observed OR among 1 1 compared with 2*2 from this meta-analysis was 1.88 (95% CI, 1.00-3.57; P for interaction = 0.43). Besides showing the effectiveness of the Mendelian randomization approach, these findings support the theory that alcohol increases head and neck cancer risk through the carcinogenic action of acetaldehyde. PMID- 19124504 TI - Two estrogen-related variants in CYP19A1 and endometrial cancer risk: a pooled analysis in the Epidemiology of Endometrial Cancer Consortium. AB - Common variants in CYP19A1 (the A alleles of rs749292 and rs727479) have been associated with a 10% to 20% increase in circulating estrogen levels in postmenopausal women. We hypothesized that the presence of one or both A alleles in these single nucleotide polymorphisms (SNP) is associated with increased endometrial cancer risk. We tested this hypothesis in a large pooled analysis of 4,998 endometrial cancer cases and 8,285 controls from 10 studies in the Epidemiology of Endometrial Cancer Consortium. The majority of women (>66%) were whites, with smaller proportions of other races and ethnic groups (blacks, Asians, and Latinas) also included in this pooled analysis. Unconditional logistic regression was used to model the association between SNPs/haplotypes and endometrial cancer risk. Carrying the A allele of either of these SNPs was associated with an increased risk of endometrial cancer, with pooled odds ratios per allele of 1.14, 95% confidence interval of 1.09-1.21, and P = 7.1 x 10(-7) for rs749292, and odds ratio per allele of 1.08, 95% confidence interval of 1.02 1.14, and P = 0.009 for rs727479. For rs749292, these associations were generally stronger among women age >or=55 years. For both SNPs, risk increased with increasing body mass index, and for rs727479, this pattern seemed stronger among women age >or=55 years (P interaction = 0.007). The combination of A alleles in the two SNPs, either by direct count or by haplotype analysis, did not increase risk above that observed for the individual SNPs. Our study provides evidence that CYP19A1 genetic variation influences susceptibility to endometrial cancer, particularly among older and obese women. PMID- 19124506 TI - Common genetic variation in candidate genes and susceptibility to subtypes of breast cancer. AB - Association studies have been widely used to search for common low-penetrance susceptibility alleles to breast cancer in general. However, breast cancer is a heterogeneous disease and it has been suggested that it may be possible to identify additional susceptibility alleles by restricting analyses to particular subtypes. We used data on 710 single nucleotide polymorphisms (SNP) in 120 candidate genes from a large candidate gene association study of up to 4,470 cases and 4,560 controls to compare the results of analyses of "overall" breast cancer with subgroup analyses based on the major clinicopathologic characteristics of breast cancer (stage, grade, morphology, and hormone receptor status). No SNP was highly significant in overall effects analysis. Subgroup analysis resulted in substantial reordering of ranks of SNPs, as assessed by the magnitude of the test statistics, and some associations that were not significant for an overall effect were detected in subgroups at a nominal 5% level adjusted for multiple testing. The most significant association of CCND1 SNP rs3212879 with estrogen receptor-negative tumor types (P = 0.001) did not reach genome-wide significance levels. These results show that it may be possible to detect associations using subgroup analysis that are missed in overall effects analysis. If the associations we found can be replicated in independent studies, they may provide important insights into disease mechanisms in breast cancer. PMID- 19124507 TI - A prospectively measured serum biomarker for a tobacco-specific carcinogen and lung cancer in smokers. AB - BACKGROUND: No prior studies have related a tobacco-specific carcinogen to the risk of lung cancer in smokers. Of the over 60 known carcinogens in cigarette smoke, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is specific to tobacco and causes lung cancer in laboratory animals. Its metabolites, 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides (total NNAL), have been studied as biomarkers of exposure to NNK. We studied the relation of prospectively measured NNK biomarkers to lung cancer risk. METHODS: In a case control study nested in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, we randomly selected 100 lung cancer cases and 100 controls who smoked at baseline and analyzed their baseline serum for total NNAL, cotinine, and r-1,t-2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene (PheT), a biomarker of polycyclic aromatic hydrocarbon exposure and metabolic activation. To examine the association of the biomarkers with all lung cancers and for histologic subtypes, we computed odds ratios for total NNAL, PheT, and cotinine using logistic regression to adjust for potential confounders. FINDINGS: Individual associations of age, smoking duration, and total NNAL with lung cancer risk were statistically significant. After adjustment, total NNAL was the only biomarker significantly associated with risk (odds ratio, 1.57 per unit SD increase; 95% confidence interval, 1.08-2.28). A similar statistically significant result was obtained for adenocarcinoma risk, but not for nonadenocarcinoma. CONCLUSIONS: This first reporting of the effect of the prospectively measured tobacco-specific biomarker total NNAL, on risk of lung cancer in smokers provides insight into the etiology of smoking-related lung cancer and reinforces targeting NNK for cancer prevention. PMID- 19124508 TI - Methionine synthase A2756G polymorphism interacts with alcohol and folate intake to influence the risk of colorectal adenoma. AB - Genomic DNA hypomethylation has been associated with colorectal carcinogenesis. Methionine synthase A2756G (MTR A2756G) is a common nonsynonymous polymorphism in the gene that encodes methionine synthase, a key enzyme in the pathway leading to DNA methylation. Several studies, but not all, have reported relatively lower plasma homocysteine among individuals with the AG or GG genotype. Meanwhile, higher plasma homocysteine was associated with genomic DNA hypomethylation in healthy volunteers. We therefore hypothesized that minor allele carriers possess a decreased risk of colorectal adenoma, and examined this hypothesis in a case control study of colorectal adenoma in Japan involving 723 cases and 670 controls. An unconditional logistic regression model was used to estimate odds ratios (OR) and their 95% confidence intervals (95% CI) for colorectal adenoma after adjustment for potential confounders. Despite the lack of an overall association, we observed a significant interaction between MTR A2756G and alcohol intake (P for interaction = 0.007). Compared with never drinkers with the AA genotype, never drinkers with the AG or GG genotype exhibited a significantly decreased risk (OR, 0.56; 95% CI, 0.34-0.90) whereas heavy drinkers with the same genotypes showed a substantially increased risk (OR, 1.90; 95% CI, 1.04-3.46). In addition, a marginally significant interaction was observed with folate intake (P for interaction = 0.07). The G allele may confer protection against colorectal adenoma in the presence of a considerably good folate status. Our findings add to increasing evidence that DNA methylation plays an important role even at an early stage of colorectal carcinogenesis. PMID- 19124509 TI - Postmenopausal levels of endogenous sex hormones and risk of colorectal cancer. AB - Observational epidemiologic studies and randomized trials have reported a protective effect of oral hormonal replacement therapy on risk of colorectal cancer. Only one previous prospective study, the Women's Health Initiative Observational Study, has reported on the relationship between endogenous hormones and incident colorectal cancer. Contrary to expectation, the investigators found that women with higher circulating estradiol levels were at increased risk of developing colorectal cancer. We conducted a case-control study nested within the New York University Women's Health Study prospective cohort to evaluate the association between endogenous levels of estrone, estradiol, and sex hormone binding globulin (SHBG) with risk of colorectal cancer. We measured hormones and SHBG in serum samples collected at enrollment from a total of 148 women who subsequently developed colorectal cancer and 293 matched controls. Circulating estrone levels were positively associated with risk of colorectal cancer: The odds ratio for the highest versus lowest quartile of estrone was 1.8 (95% confidence interval, 1.0-3.3). We found a nonsignificant inverse association between SHBG and colorectal cancer, which disappeared after adjusting for body mass index. We did not find an association between estradiol and colorectal cancer risk, but we cannot rule out a potential association because of substantial laboratory error in the measurement. Our results suggest that endogenous estrone is associated with increased risk of colorectal cancer in postmenopausal women. PMID- 19124510 TI - Insulin-like growth factor-1- and interleukin-6-related gene variation and risk of multiple myeloma. AB - Insulin-like growth factor (IGF)-1 and interleukin (IL)-6 promote the proliferation and survival of multiple myeloma cells. Variation in genes related to IGF-1 and IL-6 signaling may influence susceptibility to multiple myeloma. To assess their etiologic role, we examined the association of 70 tagging single nucleotide polymorphisms (SNP) in seven IGF-1 and three IL-6 pathway genes with multiple myeloma risk in two prospective cohorts, the Nurses' Health Study and the Health Professionals Follow-up Study. Among the participants who provided DNA specimens, we identified 58 women and 24 men with multiple myeloma and matched two controls per case. We used multivariable logistic regression models to assess the association of the SNPs or tagged haplotypes with multiple myeloma risk. Several SNPs had suggestive associations with multiple myeloma based on large odds ratios (OR), although the corresponding omnibus P values were not more than nominally significant (i.e., at P < 0.05). These SNPs included rs1801278 in the gene encoding insulin receptor substrate-1 [IRS1; C/T versus C/C genotypes; OR, 4.3; 95% confidence interval (CI), 1.5-12.1] and three IL-6 receptor SNPs: rs6684439 (T/T versus C/C; OR, 2.9; 95% CI, 1.2-7.0), rs7529229 (C/C versus T/T; OR, 2.5; 95% CI, 1.1-6.0), and rs8192284 (C/C versus A/A; OR, 2.5, 95% CI, 1.1 6.0). Additional SNPs in genes encoding IGF-1, IGF binding protein-2, IRS2, and gp130 also showed suggestive associations with multiple myeloma risk. We conducted a large number of statistical tests, and the findings may be due to chance. Nonetheless, the data are consistent with the hypothesis that IGF-1- and IL-6-related gene variation influences susceptibility to multiple myeloma and warrant confirmation in larger populations. PMID- 19124511 TI - Physical activity and postmenopausal breast cancer risk in the NIH-AARP diet and health study. AB - BACKGROUND: Although physical activity has been associated with reduced breast cancer risk, whether this association varies across breast cancer subtypes or is modified by reproductive and lifestyle factors is unclear. METHODS: We examined physical activity in relation to postmenopausal breast cancer risk in 182,862 U.S. women in the NIH-AARP Diet and Health Study. Physical activity was assessed by self-report at baseline (1995-1996), and 6,609 incident breast cancers were identified through December 31, 2003. Cox regression was used to estimate the relative risk (RR) and 95% confidence interval (95% CI) of postmenopausal breast cancer overall and by tumor characteristics. Effect modification by select reproductive and lifestyle factors was also explored. RESULTS: In multivariate models, the most active women experienced a 13% lower breast cancer risk versus inactive women (RR, 0.87; 95% CI, 0.81-0.95). This inverse relation was not modified by tumor stage or histology but was suggestively stronger for estrogen receptor (ER)-negative (RR, 0.75; 95% CI, 0.54-1.04) than ER-positive (RR, 0.97; 95% CI, 0.84-1.12) breast tumors and was suggestively stronger for overweight/obese (RR, 0.86; 95% CI, 0.77-0.96) than lean (RR, 0.95; 95% CI, 0.87 1.05) women. The inverse relation with physical activity was also more pronounced among women who had never used menopausal hormone therapy and those with a positive family history of breast cancer than their respective counterparts. CONCLUSIONS: Physical activity was associated with reduced postmenopausal breast cancer risk, particular to ER-negative tumors. These results, along with heterogeneity in the physical activity-breast cancer relation for subgroups of menopausal hormone therapy use and adiposity, indicate that physical activity likely influences breast cancer risk via both estrogenic and estrogen-independent mechanisms. PMID- 19124512 TI - Vitamin D receptor polymorphisms and breast cancer risk: results from the National Cancer Institute Breast and Prostate Cancer Cohort Consortium. AB - BACKGROUND: Vitamin D is hypothesized to lower the risk of breast cancer by inhibiting cell proliferation via the nuclear vitamin D receptor (VDR). Two common single nucleotide polymorphisms (SNP) in the VDR gene (VDR), rs1544410 (BsmI), and rs2228570 (FokI), have been inconsistently associated with breast cancer risk. Increased risk has been reported for the FokI ff genotype, which encodes a less transcriptionally active isoform of VDR, and reduced risk has been reported for the BsmI BB genotype, a SNP in strong linkage disequilibrium with a 3'-untranslated region, which may influence VDR mRNA stability. METHODS: We pooled data from 6 prospective studies in the National Cancer Institute Breast and Prostate Cancer Cohort Consortium to examine associations between these SNPs and breast cancer among >6,300 cases and 8,100 controls for each SNP using conditional logistic regression. RESULTS: The odds ratio (OR) for the rs2228570 (FokI) ff versus FF genotype in the overall population was statistically significantly elevated [OR, 1.16; 95% confidence interval (95% CI), 1.04-1.28] but was weaker once data from the cohort with previously published positive findings were removed (OR, 1.10; 95% CI, 0.98-1.24). No association was noted between rs1544410 (BsmI) BB and breast cancer risk overall (OR, 0.98; 95% CI, 0.89-1.09), but the BB genotype was associated with a significantly lower risk of advanced breast cancer (OR, 0.74; 95% CI, 0.60-0.92). CONCLUSIONS: Although the evidence for independent contributions of these variants to breast cancer susceptibility remains equivocal, future large studies should integrate genetic variation in VDR with biomarkers of vitamin D status. PMID- 19124513 TI - Randomized controlled trial of aerobic exercise on insulin and insulin-like growth factors in breast cancer survivors: the Yale Exercise and Survivorship study. AB - BACKGROUND: High insulin and insulin-like growth factor-I (IGF-I) levels may be associated with an increased breast cancer risk and/or death. Given the need to identify modifiable factors that decrease insulin, IGF-I, and breast cancer risk and death, we investigated the effects of a 6-month randomized controlled aerobic exercise intervention versus usual care on fasting insulin, IGF-I, and its binding protein (IGFBP-3) in postmenopausal breast cancer survivors. METHODS: Seventy-five postmenopausal breast cancer survivors were identified from the Yale New Haven Hospital Tumor Registry and randomly assigned to an exercise (n = 37) or usual care (n = 38) group. The exercise group participated in 150 minutes per week of moderate-intensity aerobic exercise. The usual care group was instructed to maintain their current physical activity level. A fasting blood sample was collected on each study participant at baseline and 6 months. Blood levels of insulin and IGF were measured with ELISA. RESULTS: On average, exercisers increased aerobic exercise by 129 minutes per week compared with 45 minutes per week among usual care participants (P < 0.001). Women randomized to exercise experienced decreases in insulin, IGF-I, and IGFBP-3, whereas women randomized to usual care had increases in these hormones. Between-group differences in insulin, IGF-I, and IGFBP-3 were 20.7% (P = 0.089), 8.9% (P = 0.026), and 7.9% (P = 0.006), respectively. CONCLUSIONS: Moderate-intensity aerobic exercise, such as brisk walking, decreases IGF-I and IGFBP-3. The exercise-induced decreases in IGF may mediate the observed association between higher levels of physical activity and improved survival in women diagnosed with breast cancer. PMID- 19124514 TI - Urinary isothiocyanates; glutathione S-transferase M1, T1, and P1 polymorphisms; and risk of colorectal cancer: the Multiethnic Cohort Study. AB - Although an association between diet, especially cruciferous vegetables, and colorectal cancer has been hypothesized, recent studies have been inconsistent with their findings. One possibility for the discrepant results is that the interaction with related genes has not generally been considered. The present study examined the associations among urinary isothiocyanates, glutathione S transferase (GST) polymorphisms, and colorectal cancer risk in a case-control study nested within the Multiethnic Cohort Study, based in Hawaii and Los Angeles, California. We measured prediagnositic urinary isothiocyanate levels adjusted for creatinine and analyzed GSTM1, GSTT1, and GSTP1 polymorphisms in 173 cases and 313 matched controls, with biospecimens collected between 2001 and 2006. Conditional logistic regression was used to compute odds ratios and 95% confidence intervals (95% CI). A detectable amount of urinary isothiocyanates was associated with a 41% decrease in colorectal cancer risk (95% CI, 0.36-0.98). No significant, main-effect associations were seen with a homozygous deletion of the GSTM1 or GSTT1 polymorphism, or with the AG or GG genotypes for GSTP1 rs1695. There was a weak suggestion that for individuals with the GSTP1 AG or GG genotype, a detectable amount of isothiocyanates further decreases one's risk of colorectal cancer compared with those with the GSTP1 AA genotype, but the interaction term was not statistically significant (P = 0.09). This is only the second study published on the association between urinary isothiocyanates and colorectal cancer risk. The results suggest that further studies, with larger numbers, examining a possible interaction with the GSTP1 polymorphisms are warranted. PMID- 19124515 TI - Prevaccination distribution of human papillomavirus types in women attending at cervical cancer screening in Belgium. AB - INTRODUCTION: Before the introduction of vaccination against human papillomaviruses (HPV) as a new strategy of combating cervical cancer, it is required to describe the baseline prevalence of HPV infection as well as the distribution of the different HPV types in the population and among women with cervical lesions. MATERIALS AND METHODS: Approximately 10,000 liquid cervical cell samples from women, resident of Flanders (North Belgium) and participating in cervical cancer screening, were assessed cytologically and virologically with a multiplex real-time PCR using primers targeting the E6/E7 genes of 16 HPV types. Correlations of HPV infection with age, geographic area, and occurrence of cytologic lesions were assessed. RESULTS: The prevalence of cytologic abnormalities was atypical squamous cells of undetermined significance (ASC-US), 1.6%; atypical glandular cells (AGC), 0.2%; low-grade squamous intraepithelial lesion (LSIL), 2.6%; atypical squamous cells, HSIL cannot be excluded (ASC-H), 0.3%; and high-grade squamous intraepithelial lesion (HSIL), 1.2%. The frequency of high-risk HPV infections was 11% in women without cytologic abnormalities, 77% in ASC-US, 32% in AGC, 85% in LSIL, and 93% in ASC-H and HSIL. The prevalence of high-risk HPV infection was highest in women of ages 20 to 24 years (29%) and decreased progressively with age. The percentage of women with HSIL in the entire study population attributable to infection with a particular type (AR(pop) %) was highest for HPV16 (32%), followed by HPV31 (22%), HPV39 (11%), and HPV52 (11%). HPV18 was responsible for 7% of the HSIL lesions. Elimination of HPV16 and HPV18 is expected to reduce the prevalence of ASCUS with 24%, AGC with 19%, LSIL with 29%, ASC-H with 31% and HSIL with 37%. DISCUSSION: Compared to other West European studies, the prevalence of HPV infection was considerably higher in cytologically negative women but similar in women with cervical lesions. These differences could be due to the use of a PCR with high analytic sensitivity. These data are relevant for estimating the expected and theoretical levels of vaccine protection offered as vaccinated girls gradually age into the groups from which our observations stem. Further periodic laboratory-based surveys, including genotyping of cervical cell samples and linkage with vaccine registries, are an important resource to address pending questions of the effect of HPV vaccination. Research is warranted to disentangle the causal role of individual HPV types in case of multiple infections. PMID- 19124516 TI - Body composition, abdominal fat distribution, and prostate-specific antigen test results. AB - BACKGROUND: There are competing theories explaining the observed inverse association between obesity and screening prostate-specific antigen (PSA) levels: (a) endocrine disturbances related to abdominal obesity influence PSA production and (b) increased plasma volume associated with obesity dilutes PSA. Under the endocrine disturbance hypothesis, fat mass, but not lean mass, and an abdominal distribution of fat are expected to be inversely associated with PSA levels. Under plasma volume dilution theory, PSA levels are inversely associated with both lean and fat mass and are independent of body fat distribution patterns. METHODS: Data on weight, percent body fat measured by bioimpedance, and waist circumference from approximately 8,000 men undergoing routine PSA screening were evaluated. Multivariate linear regression analyses controlling for age and race/ethnicity were used to determine whether fat, lean mass, or waist circumference were associated with PSA. RESULTS: PSA levels were significantly inversely associated with both lean and fat mass. In separate models, a 5-pound difference in lean mass and fat mass was associated, respectively with a -0.9% (P < 0.001) and -0.7% (P = 0.001) difference in PSA test results. In a model that simultaneously considered lean and fat mass, a 5-pound difference in lean mass and fat mass was associated, respectively, with a -0.6% (P = 0.03) and -0.4% (P = 0.08) difference in PSA test results. Controlling for body mass index, a 1-inch difference in waist circumference was associated with a +0.9% (P = 0.01) difference in PSA levels. CONCLUSION: The results are more consistent with predictions arising from the volume dilution theory than the hormone disturbance theory. PMID- 19124517 TI - Menstrual and reproductive factors and risk of renal cell cancer in the Multiethnic Cohort. AB - A relationship between female reproductive and menstrual factors, including exogenous hormone use, and renal cell cancer (RCC) has been hypothesized, but supporting epidemiologic evidence is limited and inconsistent. Here, the association of reproductive and menstrual factors with RCC risk was examined among 106,036 Hawaii-Los Angeles Multiethnic Cohort female participants who entered the cohort between 1993 and 1996. During an average 10.6 years of follow up, 229 RCC cases were identified among these women. Data on known and potential risk factors were obtained from the baseline questionnaire. Relative risks and 95% confidence intervals for RCC associated with each factor were estimated using Cox proportional hazard models stratified by race/ethnicity, study center, and menopausal status and adjusted for age and several confounding factors. We found no evidence of association between RCC and parity, age at first birth, age at menarche, age and type of menopause (hysterectomy or bilateral oophorectomy), use and duration of oral contraceptive, and type and duration of postmenopausal hormone use. Our results do not support the hypothesis that hormone-related factors play an etiologic role in RCC among women. PMID- 19124518 TI - Tea consumption and risk of breast cancer. AB - OBJECTIVE: The purpose of our study was to examine the association of regular tea consumption with the risk of breast cancer in a large population-based case control study from the United States. METHODS: Five thousand and eighty-two women with incident breast cancer between the ages of 20 and 74 years old from population-based cancer registries in Wisconsin, Massachusetts, and New Hampshire and 4,501 age-matched controls from lists of licensed drivers and Medicare beneficiaries completed a structured telephone interview that included information on usual tea consumption 5 years prior to the interview and other breast cancer risk factors. Logistic regression was used to obtain covariate adjusted odds ratios and 95% confidence intervals associated with quantities of tea consumed. RESULTS: Tea consumption was not related to breast cancer risk overall (P for trend = 0.18). However, when stratified by age, an inverse association was observed among women less than 50 years: those consuming three or more cups per day had a 37% reduced breast cancer risk when compared with women reporting no tea consumption (age and study site-adjusted odds ratios, 0.63; 95% confidence intervals, 0.44-0.89; P = 0.01) with a significant test for trend (P = 0.01). The inverse association noted among younger women was consistent for in situ and invasive breast cancer, and for ductal and lobular breast cancer. All results were unchanged after adjustment for established risk factors. CONCLUSION: We observed evidence to support a potential beneficial influence for breast cancer associated with moderate levels of tea consumption (three or more cups per day) among younger women. Further research is needed to confirm this association. PMID- 19124519 TI - DNA repair gene variants associated with benign breast disease in high cancer risk women. AB - Benign breast disease (BBD) is a risk factor for breast cancer and may have a heritable component. Deficient DNA repair has been implicated in breast cancer etiology and may exert its effect before BBD, a known precursor. The association between allelic variants in DNA repair genes and BBD was examined in a cohort of women in Washington County, Maryland. BBD was defined by two criteria: (a) a physician diagnosis of BBD or fibrocystic disease and/or (b) a benign breast biopsy. 3,212 women without BBD at baseline were genotyped for 12 candidate single nucleotide polymorphisms in seven DNA repair genes. Of these women, 482 subsequently reported a diagnosis of BBD. The Cox model was used to calculate hazard ratios (HR). Variant alleles of XRCC1 Arg(194)Trp (rs1799782) and ERCC4 Arg(415)Gln (rs1800067) were significantly associated with BBD [HR, 1.36; 95% confidence interval (95% CI), 1.06-1.74 and HR, 1.39; 95% CI, 1.09-1.76, respectively]. Similar estimates were also observed for each of the BBD criterion used. The BBD association for ERCC4 was even stronger among women with a family history of breast cancer (HR, 2.68; 95% CI, 1.52-4.66; P(interaction) = 0.02). This study suggests that variant alleles in DNA repair genes may modify BBD risk, a potential intermediate marker of breast cancer risk, particularly among high risk subgroups. PMID- 19124520 TI - Physical activity and ovarian cancer risk: the European Prospective Investigation into Cancer and Nutrition. PMID- 19124521 TI - Long-term alcohol consumption and risk of endometrial cancer incidence: a prospective cohort study. AB - Alcohol consumption has been hypothesized to increase the risk of endometrial cancer. We used data from the prospective population-based Swedish Mammography Cohort including 61,226 women to examine the association between alcohol and endometrial cancer incidence. Alcohol consumption was assessed with validated food frequency questionnaires at baseline 1987 to 1990 and at follow-up in 1997. During a mean follow-up of 17.6 years, 687 endometrial cancer cases were identified in the Swedish cancer registries. We found no association between alcohol consumption and endometrial cancer risk after adjustment for age, body mass index, and smoking. The multivariable rate ratios (95% confidence intervals) for the three upper categories of long-term alcohol consumption as compared with no consumption were 1.01 (0.84-1.22) for <3.4 g/d, 1.01 (0.80-1.27) for 3.4 to 9.9 g/d, and 1.09 (0.71-1.67) for >or=10 g/d, respectively. The association did not differ by age, body mass index, folic acid intake, or postmenopausal hormone use in stratified analysis. In conclusion, our results suggest that low alcohol consumption (up to one drink per day) is unlikely to substantially influence risk of endometrial cancer. PMID- 19124522 TI - Vitamin D and cancer mortality. PMID- 19124524 TI - Clinical efficacy and safety of abatacept in methotrexate-naive patients with early rheumatoid arthritis and poor prognostic factors. AB - OBJECTIVES: To assess the efficacy and safety of abatacept in methotrexate-naive patients with early rheumatoid arthritis (RA) and poor prognostic factors. METHODS: In this double-blind, phase IIIb study, patients with RA for 2 years or less were randomly assigned 1 : 1 to receive abatacept (approximately 10 mg/kg) plus methotrexate, or placebo plus methotrexate. Patients were methotrexate-naive and seropositive for rheumatoid factor (RF), anti-cyclic citrullinated protein (CCP) type 2 or both and had radiographic evidence of joint erosions. The co primary endpoints were the proportion of patients achieving disease activity score in 28 joints (DAS28)-defined remission (C-reactive protein) and joint damage progression (Genant-modified Sharp total score; TS) at year 1. Safety was monitored throughout. RESULTS: At baseline, patients had a mean DAS28 of 6.3, a mean TS of 7.1 and mean disease duration of 6.5 months; 96.5% and 89.0% of patients were RF or anti-CCP2 seropositive, respectively. At year 1, a significantly greater proportion of abatacept plus methotrexate-treated patients achieved remission (41.4% vs 23.3%; p<0.001) and there was significantly less radiographic progression (mean change in TS 0.63 vs 1.06; p = 0.040) versus methotrexate alone. Over 1 year, the frequency of adverse events (84.8% vs 83.4%), serious adverse events (7.8% vs 7.9%), serious infections (2.0% vs 2.0%), autoimmune disorders (2.3% vs 2.0%) and malignancies (0.4% vs 0%) was comparable for abatacept plus methotrexate versus methotrexate alone. CONCLUSIONS: In a methotrexate-naive population with early RA and poor prognostic factors, the combination of abatacept and methotrexate provided significantly better clinical and radiographic efficacy compared with methotrexate alone and had a comparable, favourable safety profile. PMID- 19124525 TI - Human adipose-derived mesenchymal stem cells reduce inflammatory and T cell responses and induce regulatory T cells in vitro in rheumatoid arthritis. AB - OBJECTIVES: Adult mesenchymal stem cells were recently found to suppress effector T cell and inflammatory responses and have emerged as attractive therapeutic candidates for immune disorders. In rheumatoid arthritis (RA), a loss in the immunological self-tolerance causes the activation of autoreactive T cells against joint components and subsequent chronic inflammation. The aim of this study is to characterise the immunosuppressive activity of human adipose-derived mesenchymal stem cells (hASCs) on collagen-reactive T cells from patients with RA. METHODS: The effects of hASCs on collagen-reactive RA human T cell proliferation and cytokine production were investigated, as well as effects on the production of inflammatory mediators by monocytes and fibroblast-like synoviocytes from patients with RA. RESULTS: hASCs suppressed the antigen specific response of T cells from patients with RA. hASCs inhibited the proliferative response and the production of inflammatory cytokines by collagen activated CD4 and CD8 T cells. In contrast, the numbers of IL10-producing T cells and monocytes were significantly augmented upon hASC treatment. The suppressive activity of hASCs was cell-to-cell contact dependent and independent. hASCs also stimulated the generation of FoxP3 protein-expressing CD4(+)CD25(+) regulatory T cells, with the capacity to suppress collagen-specific T cell responses. Finally, hASCs downregulated the inflammatory response and the production of matrix degrading enzymes by synovial cells isolated from patients with RA. CONCLUSIONS: The present work identifies hASCs as key regulators of immune tolerance, with the capacity to suppress T cell and inflammatory responses and to induce the generation/activation of antigen-specific regulatory T cells. PMID- 19124526 TI - Pattern of developing the performance template. AB - BACKGROUND: The pattern of energy expenditure during sustained high-intensity exercise is influenced by several variables. Data from athletic populations suggest that a pre-exercise conceptual model, or template, is a central variable relative to controlling energy expenditure. AIMS: The aim of this study was to make systematic observations regarding how the performance template develops in fit individuals who have limited specific experience with sustained high intensity exercise (eg, time trials). METHODS: The study was conducted in four parts and involved measuring performance (time and power output) during: (A) six 3 km cycle time trials, (B) three 2 km rowing time trials, (C) four 2 km rowing time trials with a training period between trials 2 and 3, and (D) three 10 km cycle time trials. All time trials were self-paced with feedback to the subjects regarding previous performances and momentary pace. RESULTS: In all four series of time trials there was a progressive pattern of improved performance averaging 6% over the first three trials and 10% over six trials. In all studies improvement was associated with increased power output during the early and middle portions of the time trial and a progressively greater terminal rating of perceived exertion. Despite the change in the pattern of energy expenditure, the subjects did not achieve the pattern usually displayed by athletes during comparable events. CONCLUSIONS: This study concludes that the pattern of learning the performance template is primarily related to increased confidence that the trial can be completed without unreasonable levels of exertion or injury, but that the process takes more than six trials to be complete. PMID- 19124527 TI - A prospective cohort study on physical activity and sports-related injuries in 10 12-year-old children. AB - OBJECTIVES: To describe the incidence and severity of injuries resulting from physical education, sports and leisure time physical activity (PA) in 10-12-year old children. DESIGN: This was a prospective cohort study conducted in primary schools, with 995 children aged 10-12 years old as participants. Individual weekly exposure was estimated from baseline and follow-up questionnaires. Exposure to physical education (PE) classes was equal in all schools. An injury was recorded if it occurred during either PE class, leisure time PA, or sports, and caused the child to at least stop the current activity. Injuries were reported within 1 week of injury onset. The main outcome measure was injury incidence density. RESULTS: During the school year a total of 119 injuries were reported by 104 children, resulting in an overall injury incidence density (ID) of 0.48 per 1000 h of exposure (95% CI 0.38 to 0.57). Injury ID was lowest for leisure time PA, followed by PE and sports, respectively. Of all injuries, 40% required medical treatment and 14% resulted in 1 or more days of absence from regular school activities. In general for girls a higher injury ID was reported than for boys, mainly caused by a twofold higher risk during leisure time PA. CONCLUSIONS: Next to specific areas of preventive interest it was found that in this specific age group, girls require special attention as they seem to be at higher injury risk than boys. PMID- 19124528 TI - Adolescent exercise associated with long-term superior measures of bone geometry: a cross-sectional DXA and MRI study. AB - OBJECTIVE: To investigate whether childhood sports participation, particularly weight-bearing sports, has any effect on bone mineral content (BMC), areal bone mineral density (aBMD) and bone geometric characteristics in middle-aged postmenopausal women. Design/ SETTING: In this cross-sectional comparison of two groups, 46 middle-aged women (mean age, 60.2 (SD 5.6) years; range, 52-73 years) were grouped according to sport participation during growth: weight-bearing sports, including high-impact weight-bearing activities; and low-impact non weight-bearing sports or no participation. MAIN OUTCOME MEASURES: Dual energy X ray absorptiometry (DXA)-measured BMC, aBMD in the lumbar spine and femur. Magnetic resonance imaging (MRI) determined bone geometric characteristics in the femur, such as femoral mid-diaphyseal cross-sectional area, periosteal and endosteal perimeters and maximum and minimum second moment of area. RESULTS: Postmenopausal middle-aged women with participation in weight-bearing sports during junior high to high school (12-18 years old) displayed significantly greater BMC in both lumbar spine and femoral neck regions, and also significantly greater femoral mid-diaphyseal bone cross-sectional area, periosteal perimeter and maximum and minimum second moment of area than the non-weight-bearing sports group. CONCLUSIONS: Adolescent weight-bearing exercise exerts preservational effects on femoral mid-diaphyseal size and shape, while DXA-measured BMC effectively identified the same tendency. Weight-bearing exercise in youth affects bone, and these effects may be preserved as BMC, geometric and structural advantages even after 40 years. PMID- 19124529 TI - Recurrent falls: pace now or wait for another fall? PMID- 19124530 TI - Pacing in elderly recurrent fallers with carotid sinus hypersensitivity: a randomised, double-blind, placebo controlled crossover trial. AB - OBJECTIVES: While carotid sinus syndrome (CSS) is traditionally defined by the association of carotid sinus hypersensitivity (CSH) with syncope, uncertainty remains over the role, if any, of complex pacing in patients with CSH and unexplained or recurrent falls. We sought to clarify the role of dual chamber pacing in this patient group in the first placebo-controlled study in CSH. DESIGN: Randomised, double-blind, crossover, placebo-controlled trial. SETTING: Specialist falls and syncope facility. PATIENTS: Consecutive subjects aged over 55 years with CSH as the sole attributable cause of three or more unexplained falls in the 6 months preceding enrolment. INTERVENTION: Dual-chamber permanent pacing with rate-drop response programming. The pacemaker was switched on (DDD/RDR) or off (ODO (placebo)) for 6 months, then crossed over to the alternate mode for a further 6 months, in randomised, double-blind fashion. MAIN OUTCOME MEASURE: The primary outcome measure was number of falls in paced and non-paced modes. RESULTS: Twenty-five of 34 subjects (mean 76.8 years (SD 9.0), 27 (79%) female) recruited completed the study. Pacing intervention had no effect on number of falls (4.04 (9.54) in DDD/RDR mode, 3.48 (7.22) in ODO; relative risk of falling in ODO mode 0.82, 95% CI 0.62 to 1.10). CONCLUSION: Permanent pacing intervention had no effect on fall rates in older patients with CSH. Further work is urgently needed to clarify the role, if any, of complex pacing in this patient group. PMID- 19124531 TI - A case report of thoracic aneurysm with aortopulmonary artery fistula. AB - Rupture of an aortic aneurysm into the pulmonary artery is a rare and often a fatal event. This complication results in the development of an acute left to right shunt, volume overload, and rapid right heart deterioration. We describe a case of thoracic aortic aneurysm in whom the diagnosis of a communication with the pulmonary artery was made on the basis of transthoracic echocardiography. PMID- 19124533 TI - TNFSF15 transcripts from risk haplotype for Crohn's disease are overexpressed in stimulated T cells. AB - TNFSF15 is a susceptibility gene for Crohn's disease (CD). It remains to be elucidated how the associated single nucleotide polymorphisms (SNPs) in TNFSF15 affect the susceptibility to CD. Because there are no non-synonymous SNPs in TNFSF15, we speculated that one or more of the SNPs associated with CD may act as cis-regulatory SNPs. To reveal the effects of the SNPs on the transcriptional activity of TNFSF15, we first examined the allelic expression imbalance of TNFSF15 in peripheral blood mononuclear cells (PBMCs). When PBMCs stimulated by phytohemagglutinin (PHA) were examined, the allelic ratio of mRNA transcribed from the risk haplotype to the non-risk haplotype increased, compared with the ratio without stimulation. When peripheral blood T cells and Jurkat cells stimulated by phorbol 12-myristate 13-acetate + ionomycin were examined, an allelic expression imbalance similar to that observed in PBMCs stimulated by PHA was confirmed. The promoter assay in stimulated Jurkat cells showed that the luciferase activity of the promoter region (-979 to +35) of the risk haplotype was significantly higher than that of the non-risk haplotype, and deletion and mutagenesis analysis demonstrated that this difference resulted from the -358T/C SNP. The promoter activity of -358C (risk allele) was higher than that of -358T (non-risk allele) in stimulated T cells. This effect of -358T/C on the transcriptional activity in stimulated T cells may confer susceptibility to CD. PMID- 19124532 TI - Adipose tissue dysfunction tracks disease progression in two Huntington's disease mouse models. AB - In addition to the hallmark neurological manifestations of Huntington's disease (HD), weight loss with metabolic dysfunction is often observed in the later stages of disease progression and is associated with poor prognosis. The mechanism for weight loss in HD is unknown. Using two mouse models of HD, the R6/2 transgenic and CAG140 knock-in mouse strains, we demonstrate that adipose tissue dysfunction is detectable at early ages and becomes more pronounced as the disease progresses. Adipocytes acquire a 'de-differentiated' phenotype characterized by impaired expression of fat storage genes. In addition, HD mice exhibit reduced levels of leptin and adiponectin, adipose tissue-derived hormones that regulate food intake and glucose metabolism. Importantly, some of these changes occur prior to weight loss and development of some of the characteristic neurological symptoms. We demonstrate that impaired gene expression and lipid accumulation in adipocytes can be recapitulated by expression of an inducible mutant huntingtin transgene in an adipocyte cell line and that mutant huntingtin inhibits transcriptional activity of the PGC-1alpha co-activator in adipocytes, which may contribute to aberrant gene expression. Thus, our findings indicate that mutant huntingtin has direct detrimental effects in cell types other than neurons. The results also indicate that circulating adipose-tissue-derived hormones may be accessible markers for HD prognosis and progression and suggest that adipose tissue may be a useful therapeutic target to improve standard of life for HD patients. PMID- 19124534 TI - Paroxysmal non-kinesigenic dyskinesia is caused by mutations of the MR-1 mitochondrial targeting sequence. AB - Paroxysmal non-kinesigenic dyskinesia (PNKD) is an autosomal-dominant movement disorder characterized by attacks of dystonia, chorea and athetosis. Myofibrillogenesis regulator-1 (MR-1), the gene responsible for PNKD, is transcribed into three alternatively spliced forms: long (MR-1L), medium (MR-1M) and small (MR-1S). Two mutations, A7V and A9V, were previously discovered in the N-terminal region common to MR-1L and MR-1S. We now found a third mutation, A33P, in a new PNKD patient in the same region. Contrary to previous reports, we show here that the mutation-free MR-1M is localized in the Golgi apparatus, ER and plasma membrane, whereas both MR-1L and MR-1S isoforms are mitochondrial proteins, imported into the organelle thanks to a 39 amino acid-long, N-terminal mitochondrial targeting sequence (MTS). The MTS, which contains all three PNKD mutations, is then cleaved off the mature proteins before their insertion in the inner mitochondrial membrane. Therefore, mature MR-1S and MR-1L of PNKD patients are identical to those of normal subjects. We found no difference in import efficiency and protein maturation between wild-type and mutant MR-1 variants. These results indicate that PNKD is due to a novel disease mechanism based on a deleterious action of the MTS. PMID- 19124535 TI - Inhibition of alpha 7-containing nicotinic ACh receptors by muscarinic M1 ACh receptors in rat hippocampal CA1 interneurones in slices. AB - Cys-loop ligand-gated nicotinic ACh receptors (nAChRs) and G protein-coupled muscarinic ACh receptors (mAChRs) are expressed on rat hippocampal interneurones where they can regulate excitability, synaptic communication and cognitive function. Even though both nAChRs and mAChRs appear to co-localize to the same interneurones, it is not clear whether there is crosstalk between them. We utilized patch-clamp techniques to investigate this issue in rat hippocampal CA1 interneurones in slices under conditions where synaptic transmission was blocked. The alpha7 nAChR-mediated currents were activated by choline, and when the activation of this receptor was preceded by the activation of the M(1) mAChR subtype, the amplitude of alpha7 responses was significantly reduced in a rapidly reversible and voltage-independent manner, without any change in the kinetics of responses. This M(1) mAChR-mediated inhibition of alpha7 nAChRs was through a PLC , calcium- and PKC-dependent signal transduction cascade. These data show that M(1) mAChRs and alpha7 nAChRs are functionally co-localized on individual rat hippocampal interneurones where the activation of these particular mAChRs inhibits alpha7 nAChR function. This information will help to understand how these cholinergic receptor systems might be regulating neuronal excitability in the hippocampus in a manner that has relevance for synaptic plasticity and cognition. PMID- 19124536 TI - Brain-derived neurotrophic factor drives the changes in excitatory synaptic transmission in the rat superficial dorsal horn that follow sciatic nerve injury. AB - Peripheral nerve injury can promote neuropathic pain. The basis of the 'central sensitization' that underlies this often intractable condition was investigated using 14-20-day chronic constriction injury (CCI) of the sciatic nerve of 20-day old rats followed by electrophysiological analysis of acutely isolated spinal cord slices. In addition, defined-medium organotypic spinal cord slice cultures were exposed for 5-6 days to brain-derived neurotrophic factor (BDNF, 200 ng ml( 1)) or to medium conditioned with activated microglia (aMCM). Since microglial activation is an early consequence of CCI, the latter manipulation allowed us to model the effect of peripheral nerve injury on the dorsal horn in vitro. Using whole-cell recording from superficial dorsal horn neurons, we found that both BDNF and CCI increased excitatory synaptic drive to putative excitatory 'radial delay' neurons and decreased synaptic excitation of inhibitory 'tonic islet/central' neurons. BDNF also attenuated synaptic excitation of putative GABAergic neurons identified by glutamic acid decarboxylase (GAD) immunoreactivity. Intrinsic neuronal properties (rheobase, input resistance and action potential discharge rates) were unaffected. Exposure of organotypic cultures to either BDNF or aMCM increased overall excitability of the dorsal horn, as seen by increased cytoplasmic Ca(2+) responses to 35 mm K(+) as monitored by confocal Fluo-4AM imaging. The effect of aMCM was attenuated by the recombinant BDNF binding protein TrkBd5 and the effect of BDNF persisted when GABAergic inhibition was blocked with SR95531. These findings suggest that CCI enhances excitatory synaptic drive to excitatory neurons but decreases that to inhibitory neurons. Both effects are mediated by nerve injury-induced release of BDNF from microglia. PMID- 19124537 TI - Parallels of snipe hunting and ROS research: the challenges of studying ROS and redox signalling in response to exercise. PMID- 19124538 TI - Passive membrane properties and electrotonic signal processing in retinal rod bipolar cells. AB - Rod bipolar cells transmit visual signals from their dendrites, where they receive input from rod photoreceptors, to their axon terminals, where they synapse onto amacrine cells. Little is known, however, about the transmission and possible transformation of these signals. We have combined axon terminal recording in retinal slices, quantitative, light-microscopic morphological reconstruction and computer modelling to obtain detailed compartmental models of rat rod bipolar cells. Passive cable properties were estimated by directly fitting the current responses of the models evoked by voltage pulses to the physiologically recorded responses. At a holding potential of -60 mV, the average best-fit parameters were 1.1 microF cm(-2) for specific membrane capacitance (C(m)), 130 Omega cm for cytoplasmic resistivity (R(i)), and 24 kOmega cm(2) for specific membrane resistance (R(m)). The passive integration of excitatory and inhibitory synaptic inputs was examined by computer modelling with physiologically realistic synaptic conductance waveforms. For both transient and steady-state synaptic inhibition, the inhibitory effect was relatively insensitive to the location of the inhibition. For transient synaptic inhibition, the time window of effective inhibition depended critically on the relative timing of inhibition and excitation. The passive signal transmission between soma and axon terminal was examined by the electrotonic transform and quantified as the frequency-dependent voltage attenuation of sinusoidal voltage waveforms. For the range of parameters explored (axon diameter and length, R(i)), the lowest cutoff frequency observed was approximately 300 Hz, suggesting that realistic scotopic visual signals will be faithfully transmitted from soma to axon terminal, with minimal passive attenuation along the axon. PMID- 19124539 TI - Involvement of ERK phosphorylation in brainstem neurons in modulation of swallowing reflex in rats. AB - In order to evaluate the neuronal mechanisms underlying functional abnormalities of swallowing in orofacial pain patients, this study investigated the effects of noxious orofacial stimulation on the swallowing reflex, phosphorylated extracellular signal-regulated kinase (pERK) and gamma-aminobutyric acid (GABA) immunohistochemical features in brainstem neurons, and also analysed the effects of brainstem lesioning and of microinjection of GABA receptor agonist or antagonist into the nucleus tractus solitarii (NTS) on the swallowing reflex in anaesthetized rats. The swallowing reflex elicited by topical administration of distilled water to the pharyngolaryngeal region was inhibited after capsaicin injection into the facial (whisker pad) skin or lingual muscle. The capsaicin induced inhibitory effect on the swallowing reflex was itself depressed after the intrathecal administration of MAPK kinase (MEK) inhibitor. No change in the capsaicin-induced inhibitory effect was observed after trigeminal spinal subnucleus caudalis lesioning, but the inhibitory effect was diminished by paratrigeminal nucleus (Pa5) lesioning. Many pERK-like immunoreactive neurons in the NTS showed GABA immunoreactivity. The local microinjection of the GABA(A) receptor agonist muscimol into the NTS produced a significant reduction in swallowing reflex, and the capsaicin-induced depression of the swallowing reflex was abolished by microinjection of the GABA(A) receptor antagonist bicuculline into the NTS. The present findings suggest that facial skin-NTS, lingual muscle NTS and lingual muscle-Pa5-NTS pathways are involved in the modulation of swallowing reflex by facial and lingual pain, respectively, and that the activation of GABAergic NTS neurons is involved in the inhibition of the swallowing reflex following noxious stimulation of facial and intraoral structures. PMID- 19124540 TI - Coupling between mechanical and neural behaviour in the human first dorsal interosseous muscle. AB - The neural drive to a muscle and its biomechanical properties determine the force at a joint. These factors may be centrally linked. We studied the relationship between the ability of first dorsal interosseous muscle (FDI) to generate index flexion force around the metacarpophalangeal joint and the neural drive it receives in a voluntary contraction. The role of FDI was assessed in two thumb postures, thumb 'down' (thumb abducted) and thumb 'up' (thumb extended), and at different thumb carpometacarpal angles. These postures were designed to change acutely the flexion moment arm for FDI. The flexion twitch force evoked by supramaximal stimulation of the ulnar nerve was measured in the two postures and the change in moment arm was assessed by ultrasonography. Subjects also made voluntary flexion contractions of the index finger of approximately 5 N in both postures during which neural drive to FDI and the long finger flexor muscles was measured using surface EMG. Recordings of FDI EMG were normalized to the maximal M wave. Five of the 15 subjects also had a radial nerve block to eliminate any co contraction of the extensor muscles, and extensor muscle EMG was monitored in subjects without radial nerve block. Compared to thumb up, flexion twitch force was approximately 60% greater, and the flexion moment arm was approximately 50% greater with the thumb down. There was minimal effect of altered carpometacarpal angle on flexion twitch force for either thumb posture. During voluntary flexion contractions, normalized FDI EMG was approximately 28% greater with thumb down, compared to thumb up, with no consistent change in neural drive to the long flexors. Hence, the contribution of FDI to index finger flexion can be altered by changes in thumb position. This is linked to changes in neural drive to FDI such that neural drive increases when the mechanical contribution increases, and provides a central mechanism to produce efficient voluntary movements. PMID- 19124541 TI - Contrary roles of kainate receptors in transmitter release at corticothalamic synapses onto thalamic relay and reticular neurons. AB - Corticothalamic fibres, which originate from layer VI pyramidal neurons in the cerebral cortex, provide excitatory synaptic inputs to both thalamic relay neurons and reticular neurons; reticular neurons in turn supply inhibitory inputs to thalamic relay neurons. Pyramidal cells in layer VI in the mouse somatosensory cortex highly express mRNA encoding kainate receptors, which facilitate or depress transmitter release at several synapses in the central nervous system. We report here that contrary modulation of transmitter release from corticothalamic fibres onto thalamic relay and reticular neurons is mediated by activation of kainate receptors in mouse thalamic ventrobasal complex and thalamic reticular nucleus. Exogenous kainate presynaptically depresses the synaptic transmission at corticothalamic synapses onto thalamic relay neurons, but facilitates it at corticothalamic synapses onto reticular neurons. Meanwhile, the lemniscal synaptic transmission, which sends primary somatosensory inputs to relay neurons, is not affected by kainate. In addition, GluR5-containing kainate receptors are involved in the depression of corticothalamic synaptic transmission onto relay neurons, but not onto reticular neurons. Furthermore, synaptically activated kainate receptors mimic these effects; high-frequency stimulation of corticothalamic fibres depresses synaptic transmission onto relay neurons, but facilitates it onto reticular neurons. Our results suggest that the opposite sensitivity of kainate receptors at the two corticothalamic synapses is governed by cortical activity and regulates the balance of excitatory and inhibitory inputs to thalamic relay neurons and therefore their excitability. PMID- 19124542 TI - Alpha-adrenergic signalling activates protein kinase D and causes nuclear efflux of the transcriptional repressor HDAC5 in cultured adult mouse soleus skeletal muscle fibres. AB - The protein kinase PKD1 has recently been linked to slow fibre-type gene expression in fast skeletal muscle through phosphorylation of class II histone deacetylase (HDAC) molecules, resulting in nuclear efflux of HDAC and consequent activation of the transcription factor MEF2. However, possible upstream activators of PKD, and the time course and signalling pathway of downstream effectors have not been determined in skeletal muscle. Using fluorescent fusion proteins HDAC5-green fluorescent protein (GFP) and PKD1-mPlum expressed in fibres isolated from predominantly slow soleus muscle and maintained for 4 days in culture, we now show that alpha-adrenergic receptor activation by phenylephrine causes a transient, PKD-dependent HDAC5-GFP nuclear efflux. Concurrent to this response, PKD1-mPlum transiently redistributes from cytoplasm to plasma membrane and nuclei, and back, during 2 h exposure to phenylephrine. The recovery may reflect alpha-receptor desensitization. In contrast, the phorbol ester PMA (phorbol-12-myristate-13-acetate, a pharmacological mimic of the downstream mediator diacylglycerol in alpha-adrenergic signalling), caused continuous PKD dependent HDAC5-GFP nuclear efflux and maintained PKD1-mPlum redistribution. In the absence of expressed HDAC, PMA increased histone H3 acetylation and increased MEF2 reporter activity in a PKD-dependent manner, consistent with PKD phosphorylation of endogenous HDAC(s) and reduced nuclear HDAC activity due to HDAC nuclear efflux. HDAC5-GFP did not respond to PMA in fibres from predominantly fast flexor digitorum brevis (FDB) muscle, but did in FDB fibres expressing exogenous PKD1. Our results demonstrate that a PKD-mediated signalling pathway for HDAC nuclear efflux is activated in slow skeletal muscle through adrenergic input, which is typically active in parallel with motor neurone input during muscular activity. PMID- 19124543 TI - Differential stimulation of myofibrillar and sarcoplasmic protein synthesis with protein ingestion at rest and after resistance exercise. AB - We aimed to determine whether there is a differential stimulation of the contractile myofibrillar and the cellular sarcoplasmic proteins after ingestion of protein and how this is affected by resistance exercise. Fasted (FAST) muscle protein synthesis was measured in seven healthy young men with a primed constant infusion of L-[ring-(13)C(6)]phenylalanine. Participants then performed an intense bout of unilateral resistance exercise followed by the consumption of 25 g of whey protein to maximally stimulate protein synthesis. In the rested (FED) leg myofibrillar (MYO) protein synthesis was elevated (P < 0.01) above FAST at 3 h (approximately 163%) but not at 1 and 5 h (P > 0.05). In contrast, MYO protein synthesis in the exercised (FED-EX) leg was stimulated above FAST at 1, 3 and 5 h (approximately 100, 216, and 229%, respectively; P < 0.01) with the increase at 5 h being greater than FED (P < 0.01). Thus, the synthesis of muscle contractile proteins is stimulated by both feeding and resistance exercise early (1 h) but has a greater duration and amplitude after resistance exercise. Sarcoplasmic (SARC) protein synthesis was similarly elevated (P < 0.01) above FAST by approximately 104% at 3 h in both FED and FED-EX suggesting SARC protein synthesis is stimulated by feeding but that this response is not augmented by resistance exercise. In conclusion, myofibrillar and sarcoplasmic protein synthesis are similarly, but transiently, stimulated with protein feeding. In contrast, resistance exercise rapidly stimulates and sustains the synthesis of only the myofibrillar protein fraction after protein ingestion. These data highlight the importance of measuring the synthetic response of specific muscle protein fractions when examining the effects of exercise and nutrition. PMID- 19124544 TI - Ca2+-dependent induction of TRPM2 currents in hippocampal neurons. AB - TRPM2 is a Ca(2+)-permeable member of the transient receptor potential melastatin family of cation channels whose activation by reactive oxygen/nitrogen species (ROS/RNS) and ADP-ribose (ADPR) is linked to cell death. While these channels are broadly expressed in the CNS, the presence of TRPM2 in neurons remains controversial and more specifically, whether they are expressed in neurons of the hippocampus is an open question. With this in mind, we examined whether functional TRPM2 channels are expressed in this neuronal population. Using a combination of molecular and biochemical approaches, we demonstrated the expression of TRPM2 transcripts and proteins in hippocampal pyramidal neurons. Whole-cell voltage-clamp recordings were subsequently carried out to assess the presence of TRPM2-mediated currents. Application of hydrogen peroxide or peroxynitrite to cultured hippocampal pyramidal neurons activated an inward current that was abolished upon removal of extracellular Ca(2+), a hallmark of TRPM2 activation. When ADPR (300 microM) was included in the patch pipette, a large inward current developed but only when depolarizing voltage ramps were continuously (1/10 s) applied to the membrane. This current exhibited a linear current-voltage relationship and was sensitive to block by TRPM2 antagonists (i.e. clotrimazole, flufenamic acid and N-(p-amylcinnamoyl)anthranilic acid (ACA)). The inductive effect of voltage ramps on the ADPR-dependent current required voltage-dependent Ca(2+) channels (VDCCs) and a rise in [Ca(2+)](i). Consistent with the need for a rise in [Ca(2+)](i), activation of NMDA receptors (NMDARs), which are highly permeable to Ca(2+), was also permissive for current development. Importantly, given the prominent vulnerability of CA1 neurons to free-radical-induced cell death, we confirmed that, with ADPR in the pipette, a brief application of NMDA could evoke a large inward current in CA1 pyramidal neurons from hippocampal slices that was abolished by the removal of extracellular Ca(2+), consistent with TRPM2 activation. Such a current was absent in interneurons of CA1 stratum radiatum. Finally, infection of cultured hippocampal neurons with a TRPM2-specific short hairpin RNA (shRNA(TRPM2)) significantly reduced both the expression of TRPM2 and the amplitude of the ADPR dependent current. Taken together, these results indicate that hippocampal pyramidal neurons possess functional TRPM2 channels whose activation by ADPR is functionally coupled to VDCCs and NMDARs through a rise in [Ca(2+)](i). PMID- 19124546 TI - Diabetic ketoacidosis caused by exposure of insulin pump to heat and sunlight. PMID- 19124545 TI - Kinetic analysis of voltage-dependent potentiation and block of the glycine alpha 3 receptor by a neuroactive steroid analogue. AB - We examined the actions of a carboxylated analogue of pregnanolone ((3alpha,5beta)-20-oxopregnane-3-carboxylic acid; 3alphaCOOH5betaP) on receptors composed of glycine receptor alpha3 subunits, expressed in Xenopus oocytes. This analogue both inhibits and potentiates this receptor; potentiation increases with more negative membrane potentials while block increases with less negative membrane potentials. We used a second analogue ((3alpha,5beta)-3 hydroxymethylpregnan-20-one; 3alphaCH(2)OH5betaP) to examine the mechanism for voltage-dependent potentiation. This analogue potentiates but does not block the glycine alpha3 receptor. Steady-state responses and current relaxations following voltage jumps support the idea that the voltage dependence of potentiation indirectly arises from a voltage dependence for channel activation by glycine, rather than an intrinsic voltage dependence for potentiation. Potentiation results from a slowing of the channel deactivation rate. In the absence of steroid, at a low [glycine] current relaxations after a voltage jump show two exponential components, with a weighted average time constant of approximately 425 ms (-50 mV, 22 degrees C). The rate for channel deactivation increases at more negative potentials (e-fold per 170 mV) whereas activation decreases (e-fold per 230 mV). The probability a channel is active at a high [glycine] is greater than 0.9. The addition of 10 microM 3alphaCH(2)OH5betaP decreases the deactivation rate by 6.3-fold (-50 mV). Voltage-dependent block by 3alphaCOOH5betaP is consistent with simple open-channel block, with voltage dependence reflecting interactions of the charge on the analogue with the electrical field. Block and unblock have equal but opposite dependence on membrane potential, and the charge on 3alphaCOOH5betaP senses approximately 70% of the membrane field at the blocking site. The apparent forward rate for block, however, is very slow (2 x 10(5) m(-1) s(-1)). PMID- 19124547 TI - Evaluating cardiovascular risk assessment for asymptomatic people. PMID- 19124548 TI - Obama's victory. PMID- 19124549 TI - It sticks in our throats too. PMID- 19124550 TI - Stepping into the therapeutics void. PMID- 19124553 TI - A Milestone in Codifying the Wisdom of Traditional Oriental Medicine: TCM, Kampo, TKM, TVM-WHO International Standard Terminologies on Traditional Medicine in the Western Pacific Region. AB - The WHO published a dictionary-type book entitled 'WHO International Standard Terminologies on Traditional Medicine in the Western Pacific Region' which has a total of 3259 technical terms which have been commonly used in traditional Chinese (TCM), Japanese (Kampo), Korean (TKM) and Vietnamese (TVM) medicines. In this comprehensive guide, each term has the English expression, the original Chinese character and a concise English definition. The book covers 3106 terms from basic theories, diagnostics, diseases, various therapeutics including acupuncture and moxibustion and even the English wording of 153 titles which are considered the most important traditional medical classics published in these four countries. A prominent feature of the compilation is the codification format that assigns numbers in hundred decimal units for each category of the section. This type of coding system provides the flexibility for adding more terminologies in the future and is useful for constructing a database for the retrieval of various published scientific articles. Overall, the usage of these standard terminologies is highly desirable to deliver accurate meanings, and ultimately to avoid a variety of expressions for a single term in different scientific manuscripts on Oriental medicine. PMID- 19124552 TI - Delayed Effect of Acupuncture Treatment in OA of the Knee: A Blinded, Randomized, Controlled Trial. AB - To assess the efficacy in providing improved function and pain relief by administering 8 weeks of acupuncture as adjunctive therapy to standard care in elderly patients with OA of the knee. This randomized, controlled, blinded trial was conducted on 55 patients with OA of the knee. Forty-one patients completed the study (26 females, 15 males, mean age +/- SD 71.7 +/- 8.6 years). Patients were randomly divided into an intervention group that received biweekly acupuncture treatment (n = 28) and a control group that received sham acupuncture (n = 27), both in addition to standard therapy, for example, NSAIDS, cyclooxygenase-2 inhibitors, acetaminophen, intra-articular hyaluronic acid and steroid injections. Primary outcomes measures were changes in the Knee Society Score (KSS) knee score and in KSS function and pain ratings at therapy onset, at 8 weeks (closure of study) and at 12 weeks (1 month after last treatment). Secondary outcomes were patient satisfaction and validity of sham acupuncture. There was significant improvement in all three scores in both groups after 8 and 12 weeks compared with baseline (P < .05). Significant differences between the intervention and control groups in the KSS knee score (P = .036) was apparent only after 12 weeks. Patient satisfaction was higher in the intervention group. Adjunctive acupuncture treatment seems to provide added improvement to standard care in elderly patients with OA of the knee. Future research should determine the optimal duration of acupuncture treatment in the context of OA. PMID- 19124554 TI - Overview of recent studies on childhood leukaemia, intra-uterine growth and diet. AB - Many studies have looked at the association between birth weight and risk of acute lymphoblastic leukaemia in children, but few have been able to examine the growth by incorporating estimates of gestational age. Recent results suggest that increased growth is associated with increased risk of childhood leukaemia. Evidence is also gathering that certain dietary intakes, possibly folate, in mothers, are also related to leukaemia occurrence. If these associations are real, they may be operating through growth factor pathways. PMID- 19124555 TI - Glucocorticoids inhibit GATA-3 phosphorylation and activity in T cells. AB - Glucocorticoid (GC) immunosuppression and anti-inflammatory action involve the regulation of several transcription factors (TFs). GCs inhibit the acute production of T-helper (Th) 1 and Th2 cytokines but ultimately favor a shift toward Th2 phenotype. GCs inhibit the transcriptional activity of T-bet Th1 TF by a transrepression mechanism. Here we analyze GC regulation of GATA-3, the master driver of Th2 differentiation. We found that GCs inhibit GATA-3 transcriptional activity. We demonstrate that this mechanism does not involve physical interaction between the glucocorticoid receptor (GR) and GATA-3 or reduction of GATA-3 binding to DNA, as described previously for T-bet. Instead, GCs inhibit GATA-3 activity by inhibition of p38 mitogen-activated protein kinase induced GATA-3 phosphorylation. GCs also inhibit GATA-3 mRNA and protein expression. Finally, GATA-3 inhibition affects the interleukin-5 gene, a central Th2 cytokine. The IC(50) of dexamethasone is 10 nM with a maximum effect at 100 nM. All inhibitory actions were blocked by the GR antagonist RU38486 (1 uM), proving the specificity of GR action. In view of the crucial role of GATA-3 in T-cell differentiation and inflammation, we propose that the mechanism of GATA-3 inhibition compared with that in T-bet may have relevant implications in understanding and modulating the anti-inflammatory and Th-regulatory properties of GCs. PMID- 19124556 TI - GADD45a is a novel candidate gene in inflammatory lung injury via influences on Akt signaling. AB - We explored the mechanistic involvement of the growth arrest and DNA damage inducible gene GADD45a in lipopolysaccharide (LPS)- and ventilator-induced inflammatory lung injury (VILI). Multiple biochemical and genomic parameters of inflammatory lung injury indicated that GADD45a(-/-) mice are modestly susceptible to intratracheal LPS-induced lung injury and profoundly susceptible to high tidal volume VILI, with increases in microvascular permeability and bronchoalveolar lavage levels of inflammatory cytokines. Expression profiling of lung tissues from VILI-challenged GADD45a(-/-) mice revealed strong dysregulation in the B-cell receptor signaling pathway compared with wild-type mice and suggested the involvement of PI3 kinase/Akt signaling components. Western blot analyses of lung homogenates confirmed approximately 50% reduction in Akt protein levels in GADD45a(-/-) mice accompanied by marked increases in Akt ubiquitination. Electrical resistance measurements across human lung endothelial cell monolayers with either reduced GADD45a or Akt expression (siRNAs) revealed significant potentiation of LPS-induced human lung endothelial barrier dysfunction, which was attenuated by overexpression of a constitutively active Akt1 transgene. These studies validate GADD45a as a novel candidate gene in inflammatory lung injury and a significant participant in vascular barrier regulation via effects on Akt-mediated endothelial signaling. PMID- 19124557 TI - Ajulemic acid, a synthetic cannabinoid, increases formation of the endogenous proresolving and anti-inflammatory eicosanoid, lipoxin A4. AB - Ajulemic acid (AjA), a synthetic nonpsychoactive cannabinoid, and lipoxin A(4) (LXA(4)), an eicosanoid formed from sequential actions of 5- and 15-lipoxygenases (LOX), facilitate resolution of inflammation. The purpose of this study was to determine whether the ability of AjA to limit the progress of inflammation might relate to an increase in LXA(4), a known anti-inflammatory and proresolving mediator. Addition of AjA (0-30 microM) in vitro to human blood and synovial cells increased production of LXA(4) (ELISA) 2- to 5-fold. Administration of AjA to mice with peritonitis resulted in a 25-75% reduction of cells invading the peritoneum, and a 7-fold increase in LXA(4) identified by mass spectrometry. Blockade of 12/15 LOX, which leads to LXA(4) synthesis via 15-HETE production, reduced (>90%) the ability of AjA to enhance production of LXA(4) in vitro. These results suggest that AjA and other agents that increase endogenous compounds that facilitate resolution of inflammation may be useful for conditions characterized by inflammation and tissue injury. PMID- 19124558 TI - Novel peptide ligand directs liposomes toward EGF-R high-expressing cancer cells in vitro and in vivo. AB - Epidermal growth factor receptor (EGF-R) is an important target in anticancer therapy. Here we report how a novel EGF-R peptide ligand (D4: Leu-Ala-Arg-Leu-Leu Thr) is identified using a computer-aided design approach from a virtual peptide library of putative EGF-R binding peptides by screening against the EGF-R X-ray crystal structure in silico and in vitro. The selected peptide is conjugated with a polyethylene glycol (PEG) lipid, and the lipid moiety of the peptide-PEG-lipid conjugate is inserted into liposome membranes by a postmodification process. D4 peptide-conjugated liposomes are found to bind to and enter cells by endocytosis specifically and efficiently in vitro in a process apparently mediated by EGF-R high-expressing cancer cells (H1299). In vivo, the D4 peptide-conjugated liposomes are found to accumulate in EGF-R-expressing xenograft tumor tissues up to 80 h after intravenous delivery, in marked contrast to controls. These results demonstrate how structure-based peptide design can be an efficient approach to identify highly novel binding ligands against important receptors. These data could have important consequences for the development of peptide-directed drug delivery systems with engineered specificities and prolonged times of action. PMID- 19124559 TI - Nanoparticle-mediated delivery of superoxide dismutase to the brain: an effective strategy to reduce ischemia-reperfusion injury. AB - Excessive production of reactive oxygen species (ROS) after cerebral ischemia and reperfusion is implicated in brain damage through different cellular and molecular mechanisms, and it is further aggravated by impaired cellular antioxidant defense systems under ischemic conditions. Therapeutic strategies based on exogenous delivery of the native form of superoxide dismutase (SOD), a free radical scavenger, are limited because of its short half-life (approximately 6 min) in vivo and poor permeability across the blood-brain-barrier (BBB). We encapsulated SOD in biodegradable poly(D,L-lactide co-glycolide) nanoparticles (SOD-NPs) and tested their efficacy in a rat focal cerebral ischemia-reperfusion injury model. We hypothesized that localized brain delivery of SOD-NPs would sustain the protective effect of SOD by neutralizing the deleterious effects of ROS formed following ischemia-reperfusion. SOD-NPs were administered at the time of reperfusion via the intracarotid route to maximize their localization in the brain. Animals receiving SOD-NPs (10,000 U of SOD/kg) demonstrated a 65% reduction in infarct volume, whereas an equivalent dose of SOD in solution (SOD Sol) increased it by 25% over saline control (P<0.001; data at 6 h following reperfusion). Control NPs alone or mixed with SOD-Sol were ineffective in reducing infract volume, with results similar to saline control, indicating the protective effect of the encapsulated enzyme. SOD-NPs maintained BBB integrity, thereby preventing edema, reduced the level of ROS formed following reperfusion, and protected neurons from undergoing apoptosis. Animals treated with SOD-NPs demonstrated greater survival than those with saline control (75% vs. 0% at 28 days) and later regained most vital neurological functions. SOD-NPs may be an effective treatment option in conjunction with a thrombolytic agent for stroke patients. PMID- 19124560 TI - Keratin 7 promoter selectively targets transgene expression to normal and neoplastic pancreatic ductal cells in vitro and in vivo. AB - Keratin 7 is expressed in simple epithelia but is expressed at low or undetectable levels in gastrointestinal epithelial cells. In the pancreas, it is present in ductal but not in acinar cells. K7 mRNA is overexpressed in pancreatic cancers. Here we use luciferase reporter assays to analyze the tissue-specific regulatory elements of murine keratin 7 (Krt7) promoter in vitro and in vivo. All elements required for appropriate cell and tissue specificity in reporter assays are present within the Krt7 -234 bp sequence. This fragment appears more selective to pancreatic ductal cells than the Krt19 promoter. GC-rich sequences corresponding to putative Sp1, AP-2 binding sites are essential for in vitro activity. Krt7-LacZ transgenic mice were generated to analyze in vivo activity. Sequences located 1.5 or 0.25 kb upstream of the transcription initiation site drive reporter expression to ductal, but not acinar, cells in transgenic mice. LacZ mRNA was detected in the pancreas as well as in additional epithelial tissues--such as the intestine and the lung--using both promoter constructs. An AdK7Luc adenovirus was generated to assess targeting selectivity in vivo by intravenous injection to immunocompetent mice and in a xenograft model of pancreatic cancer. The -0.25 kb region showed pancreatic selectivity, high activity in pancreatic cancers, and sustained transgene expression in xenografts. In conclusion, the krt7 promoter is useful to target pancreatic ductal adenocarcinoma cells in vitro and in vivo. PMID- 19124561 TI - Premature senescence of highly proliferative endothelial progenitor cells is induced by tumor necrosis factor-alpha via the p38 mitogen-activated protein kinase pathway. AB - Senescence of endothelial cells increases with systemic aging and is thought to contribute to the development of atherosclerosis. Cell therapy with highly proliferative endothelial progenitor cells (EPCs) is an emerging therapeutic option to promote endothelial regeneration, but little is known about their senescence and their vulnerability to inflammatory stressors. We therefore studied the senescence of proliferative human EPCs and investigated the effects of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) on their senescence. Human EPCs had a significantly lower rate of senescence at baseline, compared with that of mature endothelial cells. However, EPCs up-regulated the expression of the senescence-associated cell cycle arrest protein p16(INK4a) and markedly increased measured senescence levels when exposed to chronic TNF-alpha treatment. Analysis of telomere length showed that the increases in senescence were not related to changes in telomere length. Inhibition of the p38 mitogen activated protein kinase pathway blocked the induction of p16(INK4a) and cellular senescence. In conclusion, highly proliferative EPCs have a low rate of intrinsic senescence but are vulnerable to premature senescence induction by chronic proinflammatory stimulation. These findings will lead to a better understanding of physiological endothelial regeneration as well as to targeted therapies with the aim of promoting endothelial regeneration through endothelial progenitor cells. PMID- 19124562 TI - Quantitative assessment of colonic movement between prone and supine patient positions during CT colonography. AB - This paper aims to quantify changes in colonic length and positional change between supine and prone CT colonography (CTC) studies in order to aid development of image registration techniques. CTC studies in 20 patients (10 men and 10 women) with technically adequate distension were analysed using an image analysis workstation. Spatial co-ordinates of colonic landmarks were determined in both prone and supine orientations using a three-dimensional colon model view and centreline positions. Change in the co-ordinate position of colonic segments between supine and prone scans was calculated using the superior mesenteric artery as a fixed point of reference. There was no significant difference in total colonic length for subjects between prone and supine positions, nor any significant difference overall when men were compared with women. However, significant differences between sexes for individual segments were found; the ascending colon, descending colon and rectum were significantly longer in men and the sigmoid colon was longer in women. The transverse colon was the most mobile segment during positional change, with an average displacement between supine and prone scans of 4.6 cm (standard deviation, 0.48 cm) for men and 4.1 cm (standard deviation, 0.4 cm) for women. Consistent patterns of colonic positional change between supine and prone orientations were present and were thought to be most likely the result of abdominal compression. We concluded that there is minimal variation in colonic length between prone and supine orientations. Consistent patterns of colonic displacement with patient position suggest that predictable forces act upon the colon. Understanding these forces will facilitate image registration for CT colonography. PMID- 19124563 TI - Detection of small hepatocellular carcinoma: comparison of conventional gadolinium-enhanced MRI with gadolinium-enhanced MRI after the administration of ferucarbotran. AB - We compared the diagnostic efficacy of gadolinium (Gd)-enhanced MRI with that of Gd-enhanced MRI after administration of ferucarbotran for revealing small hypervascular hepatocellular carcinomas (HCCs). 24 patients with 34 HCCs (ranging in size from 0.6-2.0 cm) underwent Gd-enhanced three-dimensional dynamic MRI followed, after an interval of 5-11 days (mean, 7 days), by Gd-enhanced three dimensional dynamic MRI after administration of ferucarbotran. The two Gd enhanced arterial-phase MRI scans were compared quantitatively by measuring the tumour-liver contrast-to-noise ratio (CNR) and qualitatively by evaluating the tumour-liver contrast using matched-pairs analysis. The tumour-liver CNR with Gd enhanced arterial-phase imaging after ferucarbotran (250.3 +/- 103.7) was higher than that with Gd-enhanced arterial-phase imaging (221.1 +/- 96.1) (p < 0.001). Matched-pairs analysis indicated that, for three lesions, the relative tumour liver contrast was slightly better with Gd-enhanced arterial-phase imaging after ferucarbotran than with conventional Gd-enhanced arterial-phase imaging; however, in the case of the remaining 31 lesions, the two images were equivalent. We concluded that, although Gd-enhanced arterial-phase imaging after ferucarbotran results in better tumour-liver CNR than Gd-enhanced arterial-phase imaging, the ability of the two techniques to reveal small hypervascular HCCs is the same. PMID- 19124564 TI - One-dimensional quantitative evaluation of peripheral lung adenocarcinoma with or without ground-glass opacity on thin-section CT images using profile curves. AB - The purpose of our investigation was to compare the usefulness of the subjective visual assessment of ground-glass opacity (GGO) with a quantitative method that used a profile curve to determine prognosis. 96 adenocarcinomas were studied. Three diameters ([D1]-[D3]) were defined for estimating the diameter of tumours on the monitor: the distance between two points was measured using software that displays a CT density profile across the tumour. One experienced and one less experienced radiologist independently evaluated the following six parameters: the three diameters [D1]-[D3]; the solid portion of total tumour in the two different ratios ([D2]/[D1], [D3]/[D1]); and the area ratio of GGO for total opacity to subjective visual evaluation. Interobserver agreement between the two radiologists of the diameters (mean bias+/- 1.96 standard deviations) was as follows: [D1], -0.7 +/- 6 mm; [D2], 0.4 +/- 4.4 mm; and [D3], -0.1 +/- 4.2 mm (Bland and Altman's method). Interobserver agreement was fair in evaluating the area ratio of GGO (kappa test, kappa = 0.309). Univariate logistic regression analysis revealed that two ratios ([D2]/[D1], [D3]/[D1]) might be significantly useful in estimating lymph node metastasis (p < 0.026), lymph duct invasion (p < 0.001) and recurrence (p < 0.015). Observation of the area ratio of GGO by an experienced radiologist would be necessary for estimating lymph node metastasis (p = 0.04) and lymph duct invasion (p < 0.001). We concluded that the ratio of solid component to total tumour, which is obtainable in a more objective and simple way using profile curves obtained by software, is a more useful method of estimating prognosis than is visual assessment. PMID- 19124565 TI - The cost effectiveness of in vivo dosimetry is not proven. AB - In vivo dosimetry is a check of the radiotherapy treatment process. Recently, it has been argued that in vivo dosimetry at the start of every patient's radiotherapy is cost effective. We have examined and extended the cost effectiveness argument and show that the model does not deliver the stated benefits. PMID- 19124566 TI - Use of high-resolution MRI of the optic nerve in Graves' ophthalmopathy. AB - The aim of this study was to test the hypothesis that optic neuropathy in Graves' disease is associated with measurable optic nerve compression. Magnetic resonance appearances of 32 normal subjects and 27 patients with Graves' disease were evaluated using T(1) weighted volume imaging with multiplanar reformats along the course of the optic nerve. The optic nerve diameter was measured at seven positions along its course. Patients with thyroid orbitopathy were evaluated clinically and categorised into those with (n = 6) and without (n = 48) optic neuropathy. The mean diameter of the optic nerve in normal subjects ranged from 2.2-5.2 mm. The average orbital nerve diameter decreased the further the distance from the globe within the orbit; however, it increased within the optic canal and in the pre-chiasmal region. Optic nerve diameter in patients with Graves' disease without neuritis was not significantly different from that of subjects with normal optic nerves. In patients with optic neuritis, the optic nerve was narrower throughout the length of the nerve but narrowing was most marked in the apex of the orbit (p<0.05) and in the pre-chiasmal intracranial optic nerve (p<0.05). The normal optic nerve has a variable but predictable diameter throughout its course. In Graves' optic neuropathy the diameter of the nerve is significantly reduced in the orbital apex and in the pre-chiasmal portion. The study supports the hypothesis and provides further evidence that the likely mechanism of Graves' ophthalmopathy is compression of the optic nerve at the apex. PMID- 19124567 TI - A large-scale multicentre study of patient skin doses in interventional cardiology: dose-area product action levels and dose reference levels. AB - For 318 patients in 8 different Belgian hospitals, the entire skin-dose distribution was mapped using a grid of 70 thermoluminescence dosimeters per patient, allowing an accurate determination of the maximum skin dose (MSD). Dose area product (DAP) values, exposure parameters and geometry, together with procedure, patient and cardiologist characteristics, were also registered. Procedures were divided into two groups: diagnostic procedures (coronary angiography) and therapeutic procedures (dilatation, stent, combined procedures (e.g. coronary angiography + dilatation + stent)). The mean value of the MSD was 0.310 Gy for diagnostic and 0.699 Gy for therapeutic procedures. The most critical projection for receiving the MSD is the LAO90 (left anterior oblique) geometry. In 3% of cases, the MSD exceeded the 2 Gy dose threshold for deterministic effects. Action levels in terms of DAP values as the basis for a strategy for follow-up of patients for deterministic radiation skin effects were derived from measured MSD and cumulative DAP values. Two DAP action levels are proposed. A first DAP action level of 125 Gy cm(2) corresponding to the dose threshold of 2 Gy would imply an optional radiopathological follow-up depending on the cardiologist's decision. A second DAP action level of 250 Gy cm(2) corresponding to the 3 Gy skin dose would imply a systematic follow-up. Dose reference levels - 71.3 Gy cm(2) for diagnostic and 106.0 Gy cm(2) for therapeutic procedures - were derived from the 75 percentile of the DAP distributions. As a conclusion, we propose that total DAP is registered in patient's record file, as it can serve to improve the follow-up of patients for radiation-induced skin injuries. PMID- 19124568 TI - Quality of portal verification radiography using EC-L film in electron beam therapy. AB - The quality of the portal verification radiographs produced using the enhanced contrast localisation (EC-L) Fast cassette-EC-L film (F-EC) combination and the EC-L Oncology cassette-EC-L film (O-EC) combination was investigated fundamentally and clinically. A computerised radiography (CR) system was used for comparison. In the clinic, portal verification radiographs produced for 22 patients with breast cancer were evaluated. The characteristic curves showed that the relative speed was 0.92 for the O-EC combination when the speed of the F-EC combination was defined to be 1, and that the average gradients were 4.76 and 4.35 for the F-EC combination and the O-EC combination, respectively. The smallest visible volumes of Burger's phantom were 50.3 mm(3), 60.8 mm(3) and 199.5 mm(3) for the F-EC combination, the O-EC combination and the CR system, respectively, at an energy of 9 MeV, and 68.4 mm(3), 74.2 mm(3) and 195 mm(3), respectively, at an energy of 12 MeV. In the clinic, both combinations at an energy of 6 MeV and the O-EC combination at 9 MeV showed very poor quality owing to underdensity. However the F-EC combination at an energy of 12 MeV and the O-EC combination at an energy of 15 MeV demonstrated a higher quality. When bremsstrahlung dose passing through the body is sufficient, the quality of portal verification radiography using EC-L film is appropriate for clinical use. PMID- 19124569 TI - Potential Irish dose reference levels for cardiac interventional examinations. AB - Collective dose for cardio-angiography studies is the highest of all non-CT radiological investigations. Deterministic effects such as erythema, epilation and ulceration being reported on patients who have undergone these procedures emphasise the importance of optimising radiation dose, whilst not compromising diagnostic efficacy. This study investigated radiation doses delivered to patients for four common types of cardiac radiological examinations: coronary angiography (CA), percutaneous coronary intervention (PCI), coronary angiograms (which were followed by an interventional procedure (CA-PCI)) and permanent pacemaker insertions (PPIs). 21 cardiac imaging suites participated in the study in 14 hospitals, representing 90% of relevant centres within Ireland. Radiation dose was monitored for 1804 adult patients using dose-area product meters. Operational and examination details, such as cardiologist grade, patient details, examination complexity and exposure factors, were recorded for each examination. Variation factors in dose between centres ranged from 2.7 to 11.1, but these factors were not higher than intrahospital variations previously recorded for other examinations within Ireland, such as chest X-ray. High-dose centres were often associated with long screening times, a high patient body mass index and complexity of the procedure. Preliminary dose reference levels (DRLs) were established using rounded third quartile values at 4200 cGy cm(2), 8400 cGy cm(2), 10,700 cGy cm(2) and 2100 cGy cm(2) for CA, PCI, CA-PCI and PPI, respectively. With these commonly performed relatively high-dose procedures, it is important that some guideline values are available to encourage optimised strategies. These proposed DRLs offer such guidance. PMID- 19124570 TI - Prion variants and species barriers among Saccharomyces Ure2 proteins. AB - As hamster scrapie cannot infect mice, due to sequence differences in their PrP proteins, we find "species barriers" to transmission of the [URE3] prion in Saccharomyces cerevisiae among Ure2 proteins of S. cerevisiae, paradoxus, bayanus, cariocanus, and mikatae on the basis of differences among their Ure2p prion domain sequences. The rapid variation of the N-terminal Ure2p prion domains results in protection against the detrimental effects of infection by a prion, just as the PrP residue 129 Met/Val polymorphism may have arisen to protect humans from the effects of cannibalism. Just as spread of bovine spongiform encephalopathy prion variant is less impaired by species barriers than is sheep scrapie, we find that some [URE3] prion variants are infectious to another yeast species while other variants (with the identical amino acid sequence) are not. The species barrier is thus prion variant dependent as in mammals. [URE3] prion variant characteristics are maintained even on passage through the Ure2p of another species. Ure2p of Saccharomyces castelli has an N-terminal Q/N-rich "prion domain" but does not form prions (in S. cerevisiae) and is not infected with [URE3] from Ure2p of other Saccharomyces. This implies that conservation of its prion domain is not for the purpose of forming prions. Indeed the Ure2p prion domain has been shown to be important, though not essential, for the nitrogen catabolism regulatory role of the protein. PMID- 19124571 TI - Membrane bound axin is sufficient for Wingless signaling in Drosophila embryos. AB - The Wingless signaling pathway controls various developmental processes in both vertebrates and invertebrates. Here I probe the requirement for nuclear localization of APC2 and Axin in the Wg signal transduction pathway during embryonic development of Drosophila melanogaster. I find that nuclear localization of APC2 appears to be required, but Axin can block signaling when tethered to the membrane. These results support the model where Axin regulates Armadillo localization and activity in the cytoplasm. PMID- 19124572 TI - The genome of Thermosipho africanus TCF52B: lateral genetic connections to the Firmicutes and Archaea. AB - Lateral gene transfers (LGT) (also called horizontal gene transfers) have been a major force shaping the Thermosipho africanus TCF52B genome, whose sequence we describe here. Firmicutes emerge as the principal LGT partner. Twenty-six percent of phylogenetic trees suggest LGT with this group, while 13% of the open reading frames indicate LGT with Archaea. PMID- 19124573 TI - Synchronous gene expression of the Yersinia enterocolitica Ysa type III secretion system and its effectors. AB - Type III secretion systems (T3SSs) are complex units that consist of many proteins. Often the proteins are encoded as a cohesive unit on virulence plasmids, but several systems have their various components dispersed around the chromosome. The Yersinia enterocolitica Ysa T3SS is such a system, where the apparatus genes, some regulatory genes, and four genes encoding secreted proteins (ysp genes) are contained in a single locus. The remaining ysp genes and at least one additional regulator are found elsewhere on the chromosome. Expression of ysa genes requires conditions of high ionic strength, neutral/basic pH, and low temperatures (26 degrees C) and is stimulated by exposure to solid surfaces. The AraC-like regulator YsaE and the dual-function chaperone/regulator SycB are required to stimulate the sycB promoter, which transcribes sycB and probably yspBCDA as well. The putative phosphorelay proteins YsrRS (located at the distal end of the ysa locus) and RcsB, the response regulator of the RcsBCD phosphorelay system, are required to initiate transcription at the ysaE promoter, which drives transcription of many apparatus genes. In this work, we sought to determine which ysp genes were coordinately regulated with the genes within the ysa locus. We found that six unlinked ysp genes responded to NaCl and required YsaE/SycB, YsrRS, and RcsB for expression. Three ysp genes had unique patterns, one of which was unaffected by all elements tested except NaCl. Thus, while the ysp genes were likely to have been acquired independently, most have acquired a synchronous regulatory pattern. PMID- 19124574 TI - Genome expression analyses revealing the modulation of the Salmonella Rcs regulon by the attenuator IgaA. AB - Intracellular growth attenuator A (IgaA) was identified as a Salmonella enterica regulator limiting bacterial growth inside fibroblasts. Genetic evidence further linked IgaA to repression of the RcsCDB regulatory system, which responds to envelope stress. How IgaA attenuates this system is unknown. Here, we present genome expression profiling data of S. enterica serovar Typhimurium igaA mutants grown at high osmolarity and displaying exacerbated Rcs responses. Transcriptome data revealed that IgaA attenuates gene expression changes requiring phosphorylated RcsB (RcsB~P) activity. Some RcsB-regulated genes, yciGFE and STM1862 (pagO)-STM1863-STM1864, were equally expressed in wild-type and igaA strains, suggesting a maximal expression at low levels of RcsB ~P. Other genes, such as metB, ypeC, ygaC, glnK, glnP, napA, glpA, and nirB, were shown for the first time and by independent methods to be regulated by the RcsCDB system. Interestingly, IgaA-deficient strains with reduced RcsC or RcsD levels exhibited different Rcs responses and distinct virulence properties. spv virulence genes were differentially expressed in most of the analyzed strains. spvA expression required RcsB and IgaA but, unexpectedly, was also impaired upon stimulation of the RcsC-->RcsD-->RcsB phosphorelay. Overproduction of either RcsB(+) or a nonphosphorylatable RcsB(D56Q) variant in strains displaying low spvA expression unveiled that both dephosphorylated RcsB and RcsB~P are required for optimal spvA expression. Taken together, our data support a model with IgaA attenuating the RcsCDB system by favoring the switch of RcsB~P to the dephosphorylated state. This role of IgaA in constantly fine-tuning the RcsB~P/RcsB ratio may ensure the proper expression of important virulence factors, such as the Spv proteins. PMID- 19124576 TI - Regions on Gli349 and Gli521 protein molecules directly involved in movements of Mycoplasma mobile gliding machinery, suggested by use of inhibitory antibodies and mutants. AB - Mycoplasma mobile glides on solid surfaces by use of a unique mechanism that involves two large proteins, Gli349 and Gli521. Here we isolated and analyzed two antibodies and three mutants that modified mycoplasma gliding. Mapping of the target points of antibodies and mutations currently available suggested that a 301-amino-acid region on the whole 3,138-amino-acid sequence, a C-terminal region of Gli349, and an N-terminal region of Gli521 are directly involved in the movements of the gliding machinery. PMID- 19124575 TI - Contact-dependent growth inhibition causes reversible metabolic downregulation in Escherichia coli. AB - Contact-dependent growth inhibition (CDI) is a mechanism identified in Escherichia coli by which bacteria expressing two-partner secretion proteins encoded by cdiA and cdiB bind to BamA in the outer membranes of target cells and inhibit their growth. A third gene in the cluster, cdiI, encodes a small protein that is necessary and sufficient to confer immunity to CDI, thereby preventing cells expressing the cdiBA genes from inhibiting their own growth. In this study, the cdiI gene was placed under araBAD promoter control to modulate levels of the immunity protein and thereby induce CDI by removal of arabinose. This CDI autoinhibition system was used for metabolic analyses of a single population of E. coli cells undergoing CDI. Contact-inhibited cells showed altered cell morphology, including the presence of filaments. Notably, CDI was reversible, as evidenced by resumption of cell growth and normal cellular morphology following induction of the CdiI immunity protein. Recovery of cells from CDI also required an energy source. Cells undergoing CDI showed a significant, reversible downregulation of metabolic parameters, including aerobic respiration, proton motive force (Deltap), and steady-state ATP levels. It is unclear whether the decrease in respiration and/or Deltap is directly involved in growth inhibition, but a role for ATP in the CDI mechanism was ruled out using an atp mutant. Consistent with the observed decrease in Deltap, the phage shock response was induced in cells undergoing CDI but not in recovering cells, based on analysis of levels of pspA mRNA. PMID- 19124577 TI - Cystathionine gamma-lyase is a component of cystine-mediated oxidative defense in Lactobacillus reuteri BR11. AB - Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H(2)S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine gamma-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H(2)S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H(2)S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. PMID- 19124578 TI - A pseudouridine synthase homologue is critical to cellular differentiation in Toxoplasma gondii. AB - Toxoplasma gondii is a haploid protozoan parasite infecting about one in seven people in the United States. Key to the worldwide prevalence of T. gondii is its ability to establish a lifelong, chronic infection by evading the immune system, and central to this is the developmental switch between the two asexual forms, tachyzoites and bradyzoites. A library of mutants defective in tachyzoite-to bradyzoite differentiation (Tbd(-)) was created through insertional mutagenesis. This library contains mutants that, compared to the wild type, are between 20% and 74% as efficient at stage conversion. Two mutants, TBD5 and TBD8, with disruptions in a gene encoding a putative pseudouridine synthase, PUS1, were identified. The disruption in TBD8 is in the 5' end of the PUS1 gene and appears to produce a null allele with a 50% defect in differentiation. This is about the same switch efficiency as obtained with an engineered pus1 deletion mutant (Deltapus1). The insertion in TBD5 is within the PUS1 coding region, and this appears to result in a more extreme phenotype of only approximately 10% switch efficiency. Complementation of TBD8 with the genomic PUS1 allele restored wild type differentiation efficiency. Infection of mice with pus1 mutant strains results in increased mortality during the acute phase and higher cyst burdens during the chronic infection, demonstrating an aberrant differentiation phenotype in vivo due to PUS1 disruption. Our results suggest a surprising and important role for RNA modification in this biological process. PMID- 19124579 TI - Deletion of the protein kinase A regulatory subunit leads to deregulation of mitochondrial activation and nuclear duplication in Aspergillus fumigatus. AB - Proper regulation of the cyclic AMP-dependent protein kinase (PKA) pathway is required for normal growth and development in many fungi. We have reported that deletion of the PKA regulatory subunit gene, pkaR, in Aspergillus fumigatus leads to defects in germination and a hypersensitivity of conidia to oxidative stress. In this study, we further analyzed the defects of DeltapkaR conidia and found that a large proportion were abnormally larger than wild type. Because swelling and increased susceptibility to oxidative stress are characteristic of germinating conidia, we analyzed the metabolic activity of the conidia by mitochondrial staining. Whereas it required 4 h in rich medium for wild-type mitochondria to become active, DeltapkaR conidia harbored active mitochondria in the absence of a germinant. Furthermore, conidia of the mutant showed a dramatic loss in viability upon short-term storage in water, indicating starvation-induced death. Taken together, our data suggest that PKA activity regulates metabolic activation of resting conidia. Additionally, the DeltapkaR mutant displayed an abnormal abundance of hyphal nuclei and had increased transcript levels of several cell cycle regulatory genes. These data indicate an important role for PKA in the nuclear duplication cycle of A. fumigatus. PMID- 19124581 TI - Brimonidine tartrate poisoning in children: frequency, trends, and use of naloxone as an antidote. AB - BACKGROUND: Brimonidine tartrate, a centrally acting selective alpha-2 adrenergic agonist with a toxicity that is often compared with that of clonidine, is used as eye drops to lower intraocular pressure in glaucoma. We investigated characteristics of childhood exposures to brimonidine-containing products. METHODS: All brimonidine exposures in children 0 to 5 years of age between 1997 and 2005 were retrieved from the American Association of Poison Control Centers' Toxic Exposure Surveillance System database and the US Food and Drug Administration's Medwatch Adverse Events Reporting System. The design of the study was retrospective, and the main outcome measures were frequency of exposures over time, reason, symptoms/signs of toxicity, dose, management site, treatment, and outcome. RESULTS: There were 413 brimonidine reports in the Toxic Exposure Surveillance System and 340 in the Adverse Events Reporting System during the 9 years under study, of which 185 Toxic Exposure Surveillance System reports involved children < or =5 years of age versus 15 cases in the Adverse Events Reporting System. There were no deaths. The peak age of poisoning in the Toxic Exposure Surveillance System pediatric cases was 2 years of age, and circumstances were unintentional poisoning in 176 cases, usually by ingestion (84.3%). Common symptoms in 176 children included drowsiness (40.9%), ataxia (4.5%), pallor (4.5%), irritability (4.0%), hypotension (4.0%), bradycardia (4.0%), miosis (3.4%), and respiratory depression (3.4%). Of the 176 unintentional pediatric poisonings, 73 children were observed at home and 103 were seen at a health care facility; 28 were hospitalized and 11 received naloxone. Of the 15 pediatric cases in the Adverse Events Reporting System data set, all were hospitalized, and 13 had ocular exposures only. CONCLUSIONS: All children < or =5 years of age with confirmed brimonidine ingestions should be medically evaluated and monitored for an extended period. Indications for the use of naloxone in brimonidine poisoning remain uncertain. PMID- 19124582 TI - CD4 counts decline despite nutritional recovery in HIV-infected Zambian children with severe malnutrition. AB - OBJECTIVE: The objective of this study was to establish the contribution that severe malnutrition makes to CD4 lymphopenia in HIV-infected and uninfected children and to determine the changes in CD4 count during nutritional rehabilitation. METHODS: Fifty-six children with severe malnutrition and with and without HIV infection were recruited from a pediatric ward in Lusaka for measurement of CD4 counts on admission, on discharge, and at final nutritional recovery. RESULTS: HIV-uninfected children with severe malnutrition had normal CD4 counts. In contrast, CD4 counts in HIV-infected children with severe malnutrition were reduced, more so in those without edema compared with those with edema. Mean CD4 count of HIV-infected SM children fell despite nutritional recovery so that at the time of full nutritional recovery, >85% of HIV-infected children required antiretroviral therapy. CONCLUSIONS: Severe malnutrition did not reduce the CD4 counts of children without HIV. HIV-infected children with severe malnutrition may respond well to nutritional rehabilitation, despite low CD4 counts, but nearly all require early antiretroviral therapy to prevent disease progression. PMID- 19124583 TI - Cotransmission in the autonomic nervous system. PMID- 19124584 TI - Eukaryotic viruses in wastewater samples from the United States. AB - Human fecal matter contains a large number of viruses, and current bacterial indicators used for monitoring water quality do not correlate with the presence of pathogenic viruses. Adenoviruses and enteroviruses have often been used to identify fecal pollution in the environment; however, other viruses shed in fecal matter may more accurately detect fecal pollution. The purpose of this study was to develop a baseline understanding of the types of viruses found in raw sewage. PCR was used to detect adenoviruses, enteroviruses, hepatitis B viruses, herpesviruses, morbilliviruses, noroviruses, papillomaviruses, picobirnaviruses, reoviruses, and rotaviruses in raw sewage collected throughout the United States. Adenoviruses and picobirnaviruses were detected in 100% of raw sewage samples and 25% and 33% of final effluent samples, respectively. Enteroviruses and noroviruses were detected in 75% and 58% of raw sewage samples, respectively, and both viral groups were found in 8% of final effluent samples. This study showed that adenoviruses, enteroviruses, noroviruses, and picobirnaviruses are widespread in raw sewage. Since adenoviruses and picobirnaviruses were detected in 100% of raw sewage samples, they are potential markers of fecal contamination. Additionally, this research uncovered previously unknown sequence diversity in human picobirnaviruses. This baseline understanding of viruses in raw sewage will enable educated decisions to be made regarding the use of different viruses in water quality assessments. PMID- 19124585 TI - Environmental poliovirus surveillance during oral poliovirus vaccine and inactivated poliovirus vaccine use in Cordoba Province, Argentina. AB - This study compares the presence of environmental poliovirus in two Argentinean populations using oral poliovirus vaccine (OPV) or inactivated poliovirus vaccine (IPV). From January 2003 to December 2005, Cordoba City used IPV in routine infant immunizations, with the exception of intermittent OPV use in August 2005. Between May 2005 and April 2006, we collected weekly wastewater samples in Cordoba City and the province's three major towns, which continued OPV use at all times. Wastewater samples were processed and analyzed for the presence of poliovirus according to WHO guidelines. During the months of IPV use in Cordoba City, the overall proportion of poliovirus-positive samples was 19%. During an intermittent switch from IPV to OPV, this proportion increased to 100% within 2 months. During the 3 months when IPV was reintroduced to replace OPV, a substantial proportion of samples (25%) remained positive for poliovirus. In the OPV-using sites, on average, 54% of samples were poliovirus positive. Seventy seven percent of poliovirus isolates showed at least one mutation in the VP1 encoding sequence; the maximum genetic divergence from the Sabin strain was 0.7%. Several isolates showed mutations on attenuation markers in the VP1-encoding sequence. The frequency or type of virus mutation did not differ between periods of IPV and OPV use or by virus serotypes. This study indicates that the sustained transmission of OPV viruses was limited during IPV use in a middle-income country with a temperate climate. The continued importation of poliovirus and genetic instability of vaccine strains even in the absence of sustained circulation suggest that high poliovirus vaccine coverage has to be maintained for all countries until the risk of reintroduction of either wild or vaccine-derived poliovirus is close to zero worldwide. PMID- 19124586 TI - Characterization of a monoclonal antibody directed against Salmonella enterica serovar Typhimurium and serovar [4,5,12:i:-]. AB - Flagellar extracts of Salmonella enterica serovars expressing phase 2 H1 antigenic complex (H:1,2, H:1,5, H:1,6, and H:1,7) and a mutant flagellin obtained by site-directed mutagenesis of the fljB gene from serovar Typhimurium at codon 218, transforming threonine to alanine, expressed in Escherichia coli (fljB218(A)) were used to analyze the H1 antigenic complex. Cross-reactions were detected by Western blotting and dot blotting using commercial polyclonal antibodies against the different wild-type extracts and mutant FljB218(A). Therefore, we produced a monoclonal antibody (MAb), 23D4, isotyped as immunoglobulin M, against H:1,2 S. enterica serovar Typhimurium flagellin. The mutant flagellin was not recognized by this MAb. When a large number of phase 1 and phase 2 flagellin antigens of different serovars were used to characterize the 23D4 MAb, only extracts of serovars Typhimurium and [4,5,12:i:-] reacted. The protein composition of phase 1 and phase 2 extracts and highly purified H:1,2 flagellin from serovar Typhimurium strain LT2 and extract of strain 286 (serovar [4,5,12:i:-]), which reacted with the MAb, was studied. Phase 2 flagellin (FljB(H:1,2)) was detected in phase 1 and phase 2 flagellar heat extracts of serovar Typhimurium and was the single protein identified in all spots of purified H:1,2 flagellin. FliC, FlgK, and other proteins were detected in some immunoreactive spots and in the flagellar extract of serovar [4,5,12:i:-]. Immunoelectron microscopy of complete bacteria with 23D4 showed MAb attachment at the base of flagella, although the MAb failed to recognize the filament of flagella. Nevertheless, the results obtained by the other immunological tests (enzyme-linked immunosorbent assay, Western blotting, and dot blotting) indicate a reaction against flagellins. The epitopes could also be shared by other proteins on spots where FljB is not present, such as aminopeptidase B, isocitrate lyase, InvE, EF-TuA, enolase, DnaK, and others. In conclusion, MAb 23D4 can be useful for detection and diagnostic purposes of S. enterica serovar Typhimurium and serovar [4,5,12:i:-] and could be also helpful for epitope characterization of flagellum-associated antigens. PMID- 19124588 TI - Simple adhesive-tape-based sampling of tomato surfaces combined with rapid fluorescence in situ hybridization for Salmonella detection. AB - A simple adhesive-tape-based method for sampling of tomato surfaces was combined with fluorescence in situ hybridization for rapid culture-independent detection of Salmonella strains. Tapes could also be placed face-down on selective agar for on-tape enrichment of captured Salmonella cells. Overlay of cell-charged tapes with small volumes of liquid enrichment media enabled subsequent detection of tape-captured Salmonella via flow cytometry. PMID- 19124587 TI - O2 and reactive oxygen species detoxification complex, composed of O2-responsive NADH:rubredoxin oxidoreductase-flavoprotein A2-desulfoferrodoxin operon enzymes, rubperoxin, and rubredoxin, in Clostridium acetobutylicum. AB - Clostridium acetobutylicum, an obligate anaerobe, grows normally under continuous O(2)-flow culture conditions, where the cells consume O(2) proficiently. An O(2) responsive NADH:rubredoxin oxidoreductase operon composed of three genes (nror, fprA2, and dsr), encoding NROR, functionally uncharacterized flavoprotein A2 (FprA2), and the predicted superoxide reductase desulfoferrodoxin (Dsr), has been proposed to participate in defense against O(2) stress. To functionally characterize these proteins, native NROR from C. acetobutylicum, recombinant NROR (rNROR), FprA2, Dsr, and rubredoxin (Rd) expressed in Escherichia coli were purified. Purified native NROR and rNROR both exhibited weak H(2)O(2)-forming NADH oxidase activity that was slightly activated by Rd. A mixture of NROR, Rd, and FprA2 functions as an efficient H(2)O-forming NADH oxidase with a high affinity for O(2) (the K(m) for O(2) is 2.9 +/- 0.4 microM). A mixture of NROR, Rd, and Dsr functions as an NADH-dependent O(2)(-) reductase. A mixture of NROR, Rd, and rubperoxin (Rpr, a rubrerythrin homologue) functions as an inefficient H(2)O-forming NADH oxidase but an efficient NADH peroxidase with a low affinity for O(2) and a high affinity for H(2)O(2) (the K(m)s for O(2) and H(2)O(2) are 303 +/- 39 microM and 95% oxygen for 4 days, followed by return to normoxia. Experimental groups received vehicle, dexamethasone, or granulocyte-macrophage colony-stimulating factor (GM-CSF). Compared with the vehicle-treated group, treatment with dexamethasone reduced Fas ligand expression and significantly improved survival. Similarly, treatment with GM-CSF, an agent we have shown protects alveolar epithelial cells against apoptosis, decreased Fas ligand expression and also improved survival. Our results suggest that the dual stresses of P. murina infection and hyperoxia induce lung injury via activation of the Fas-Fas ligand pathway and that corticosteroids and GM-CSF reduce mortality in P. murina-infected mice exposed to hyperoxic stress by inhibition of inflammation and apoptosis. PMID- 19124602 TI - Anthrax lethal toxin triggers the formation of a membrane-associated inflammasome complex in murine macrophages. AB - Multiple microbial components trigger the formation of an inflammasome complex that contains pathogen-specific nucleotide oligomerization and binding domain (NOD)-like receptors (NLRs), caspase-1, and in some cases the scaffolding protein ASC. The NLR protein Nalp1b has been linked to anthrax lethal toxin (LT)-mediated cytolysis of murine macrophages. Here we demonstrate that in unstimulated J774A.1 macrophages, caspase-1 and Nalp1b are membrane associated and part of approximately 200- and approximately 800-kDa complexes, respectively. LT treatment of these cells resulted in caspase-1 recruitment to the Nalp1b containing complex, concurrent with processing of cytosolic caspase-1 substrates. We further demonstrated that Nalp1b and caspase-1 are able to interact with each other. Intriguingly, both caspase-1 and Nalp1b were membrane associated, while the caspase-1 substrate interleukin-18 was cytosolic. Caspase-1-associated inflammasome components included, besides Nalp1b, proinflammatory caspase-11 and the caspase-1 substrate alpha-enolase. Asc was not part of the Nalp1b inflammasome in LT-treated macrophages. Taken together, our findings suggest that LT triggers the formation of a membrane-associated inflammasome complex in murine macrophages, resulting in cleavage of cytosolic caspase-1 substrates and cell death. PMID- 19124603 TI - Shiga toxin 2 targets the murine renal collecting duct epithelium. AB - Hemolytic-uremic syndrome (HUS) caused by Shiga toxin-producing Escherichia coli infection is a leading cause of pediatric acute renal failure. Bacterial toxins produced in the gut enter the circulation and cause a systemic toxemia and targeted cell damage. It had been previously shown that injection of Shiga toxin 2 (Stx2) and lipopolysaccharide (LPS) caused signs and symptoms of HUS in mice, but the mechanism leading to renal failure remained uncharacterized. The current study elucidated that murine cells of the glomerular filtration barrier were unresponsive to Stx2 because they lacked the receptor glycosphingolipid globotriaosylceramide (Gb(3)) in vitro and in vivo. In contrast to the analogous human cells, Stx2 did not alter inflammatory kinase activity, cytokine release, or cell viability of the murine glomerular cells. However, murine renal cortical and medullary tubular cells expressed Gb(3) and responded to Stx2 by undergoing apoptosis. Stx2-induced loss of functioning collecting ducts in vivo caused production of increased dilute urine, resulted in dehydration, and contributed to renal failure. Stx2-mediated renal dysfunction was ameliorated by administration of the nonselective caspase inhibitor Q-VD-OPH in vivo. Stx2 therefore targets the murine collecting duct, and this Stx2-induced injury can be blocked by inhibitors of apoptosis in vivo. PMID- 19124604 TI - The Schistosoma mansoni protein Sm16/SmSLP/SmSPO-1 assembles into a nine-subunit oligomer with potential To inhibit Toll-like receptor signaling. AB - The Sm16/SmSLP/SmSPO-1 (Sm16) protein is secreted by the parasite Schistosoma mansoni during skin penetration and has been ascribed immunosuppressive activities. Here we describe the strategy behind the design of a modified Sm16 protein with a decreased aggregation propensity, thus facilitating the expression and purification of an Sm16 protein that is soluble in physiological buffers. The Stokes radii and sedimentation coefficients of recombinant and native proteins indicate that Sm16 is an approximately nine-subunit oligomer. Analysis of truncated Sm16 derivatives showed that both oligomerization and binding to the plasma membrane of human cells depend on multiple C-terminal regions. For analysis of immunomodulatory activities, Sm16 was expressed in Pichia pastoris to facilitate the preparation of a pyrogen/endotoxin-free purified protein. Recombinant Sm16 was found to have no effect on T-lymphocyte activation, cell proliferation, or the basal level of cytokine production by whole human blood or monocytic cells. However, Sm16 exerts potent inhibition of the cytokine response to the Toll-like receptor (TLR) ligands lipopolysaccharide (LPS) and poly(I:C) while being less efficient at inhibiting the response to the TLR ligand peptidoglycan or a synthetic lipopeptide. Since Sm16 specifically inhibits the degradation of the IRAK1 signaling protein in LPS-stimulated monocytes, our findings indicate that inhibition is exerted proximal to the TLR complex. PMID- 19124606 TI - RNA-regulated interaction of transportin-1 and exportin-5 with the double stranded RNA-binding domain regulates nucleocytoplasmic shuttling of ADAR1. AB - Double-stranded RNA (dsRNA)-binding proteins interact with substrate RNAs via dsRNA-binding domains (dsRBDs). Several proteins harboring these domains exhibit nucleocytoplasmic shuttling and possibly remain associated with their substrate RNAs bound in the nucleus during nuclear export. In the human RNA-editing enzyme ADAR1-c, the nuclear localization signal overlaps the third dsRBD, while the corresponding import factor is unknown. The protein also lacks a clear nuclear export signal but shuttles between the nucleus and the cytoplasm. Here we identify transportin-1 as the import receptor for ADAR1. Interestingly, dsRNA binding interferes with transportin-1 binding. At the same time, each of the dsRBDs in ADAR1 interacts with the export factor exportin-5. RNA binding stimulates this interaction but is not a prerequisite. Thus, our data demonstrate a role for some dsRBDs as RNA-sensitive nucleocytoplasmic transport signals. dsRBD3 in ADAR1 can mediate nuclear import, while interaction of all dsRBDs might control nuclear export. This finding may have implications for other proteins containing dsRBDs and suggests a selective nuclear export mechanism for substrates interacting with these proteins. PMID- 19124605 TI - Canonical initiation factor requirements of the Myc family of internal ribosome entry segments. AB - Initiation of protein synthesis in eukaryotes requires recruitment of the ribosome to the mRNA and its translocation to the start codon. There are at least two distinct mechanisms by which this process can be achieved; the ribosome can be recruited either to the cap structure at the 5' end of the message or to an internal ribosome entry segment (IRES), a complex RNA structural element located in the 5' untranslated region (5'-UTR) of the mRNA. However, it is not well understood how cellular IRESs function to recruit the ribosome or how the 40S ribosomal subunits translocate from the initial recruitment site on the mRNA to the AUG initiation codon. We have investigated the canonical factors that are required by the IRESs found in the 5'-UTRs of c-, L-, and N-myc, using specific inhibitors and a tissue culture-based assay system, and have shown that they differ considerably in their requirements. The L-myc IRES requires the eIF4F complex and the association of PABP and eIF3 with eIF4G for activity. The minimum requirements of the N- and c-myc IRESs are the C-terminal domain of eIF4G to which eIF4A is bound and eIF3, although interestingly this protein does not appear to be recruited to the IRES RNA via eIF4G. Finally, our data show that all three IRESs require a ternary complex, although in contrast to c- and L-myc IRESs, the N-myc IRES has a lesser requirement for a ternary complex. PMID- 19124607 TI - Tropomyosin isoform expression regulates the transition of adhesions to determine cell speed and direction. AB - The balance of transition between distinct adhesion types contributes to the regulation of mesenchymal cell migration, and the characteristic association of adhesions with actin filaments led us to question the role of actin filament associating proteins in the transition between adhesive states. Tropomyosin isoform association with actin filaments imparts distinct filament structures, and we have thus investigated the role for tropomyosins in determining the formation of distinct adhesion structures. Using combinations of overexpression, knockdown, and knockout approaches, we establish that Tm5NM1 preferentially stabilizes focal adhesions and drives the transition to fibrillar adhesions via stabilization of actin filaments. Moreover, our data suggest that the expression of Tm5NM1 is a critical determinant of paxillin phosphorylation, a signaling event that is necessary for focal adhesion disassembly. Thus, we propose that Tm5NM1 can regulate the feedback loop between focal adhesion disassembly and focal complex formation at the leading edge that is required for productive and directed cell movement. PMID- 19124608 TI - Preimplantation mouse embryos depend on inhibitory phosphorylation of separase to prevent chromosome missegregation. AB - Separase is a critical protease that catalyzes the cleavage of sister chromatid cohesins to allow the separation of sister chromatids in the anaphase. Its activity must be inhibited prior to the onset of the anaphase. Two inhibitory mechanisms exist in vertebrates that block the protease activity. One mechanism is through binding and inhibition by securin, and another is phosphorylation on Ser1126 (in humans [Ser1121 in mice]). These two mechanisms are largely redundant. However, phosphorylation on Ser1121 is critical for the prevention of premature sister separation in embryonic germ cells. As a result, Ser1121-to-Ala mutation leads to depletion of germ cells in development and subsequently to infertility in mice. Here, we report that the same mutation also causes embryogenesis failure between the 8- and 16-cell stages in mice. Our results indicate a critical role of separase phosphorylation in germ cell development as well as in early embryogenesis. Thus, deregulation of separase may be a significant contributor to infertility in humans. PMID- 19124609 TI - Involvement of histone demethylase LSD1 in Blimp-1-mediated gene repression during plasma cell differentiation. AB - Plasma cell differentiation is orchestrated by the transcriptional repressor B lymphocyte-induced maturation protein-1 (Blimp-1), which silences the gene expression program of mature B cells. The molecular mechanism underlying Blimp-1 suppression of mature B-cell gene expression is not fully understood. Here we report that a proline-rich domain in Blimp-1 directly interacts with LSD1, a histone lysine demethylase. Both LSD1 knockdown and expression of Blimp-1 lacking the proline-rich domain derepressed the activities of Blimp-1-dependent luciferase reporters. Disruption of the Blimp-1 interaction with LSD1 or reduced LSD1 expression attenuated antibody production, demonstrating the biological significance of this interaction. Finally, using chromatin immunoprecipitation, we showed that Blimp-1 binding to its target sites is accompanied by LSD1 binding to those same sites and that LSD1 binding correlates with histone modifications of accessible chromatin. These findings provide further insights into the molecular mechanism of the silencing of mature B-cell genes by Blimp-1 in plasma cell differentiation. PMID- 19124610 TI - TRF1 controls telomere length and mitotic fidelity in epithelial homeostasis. AB - TRF1 is a component of the shelterin complex at mammalian telomeres; however, a role for TRF1 in telomere biology in the context of the organism is unclear. In this study, we generated mice with transgenic TRF1 expression targeted to epithelial tissues (K5TRF1 mice). K5TRF1 mice have shorter telomeres in the epidermis than wild-type controls do, and these are rescued in the absence of the XPF nuclease, indicating that TRF1 acts as a negative regulator of telomere length by controlling XPF activity at telomeres, similar to what was previously described for TRF2-overexpressing mice (K5TRF2 mice). K5TRF1 cells also show increased end-to-end chromosomal fusions, multitelomeric signals, and increased telomere recombination, indicating an impact of TRF1 on telomere integrity, again similar to the case in K5TRF2 cells. Intriguingly, K5TRF1 cells, but not K5TRF2 cells, show increased mitotic spindle aberrations. TRF1 colocalizes with the spindle assembly checkpoint proteins BubR1 and Mad2 at mouse telomeres, indicating a link between telomeres and the mitotic spindle. Together, these results demonstrate that TRF1, like TRF2, negatively regulates telomere length in vivo by controlling the action of the XPF nuclease at telomeres; in addition, TRF1 has a unique role in the mitotic spindle checkpoint. PMID- 19124611 TI - Auto- and cross-regulation of the hnRNP L proteins by alternative splicing. AB - We recently characterized human hnRNP L as a global regulator of alternative splicing, binding to CA-repeat and CA-rich elements. Here we report that hnRNP L autoregulates its own expression on the level of alternative splicing. Intron 6 of the human hnRNP L gene contains a short exon that, if used, introduces a premature termination codon, resulting in nonsense-mediated decay (NMD). This "poison exon" is preceded by a highly conserved CA-rich cluster extending over 800 nucleotides that binds hnRNP L and functions as an unusually extended, intronic enhancer, promoting inclusion of the poison exon. As a result, excess hnRNP L activates NMD of its own mRNA, thereby creating a negative autoregulatory feedback loop and contributing to homeostasis of hnRNP L levels. We present experimental evidence for this mechanism, based on NMD inactivation, hnRNP L binding assays, and hnRNP L-dependent alternative splicing of heterologous constructs. In addition, we demonstrate that hnRNP L cross-regulates inclusion of an analogous poison exon in the hnRNP L-like pre-mRNA, which explains the reciprocal expression of the two closely related hnRNP L proteins. PMID- 19124612 TI - Bezafibrate at clinically relevant doses decreases serum/liver triglycerides via down-regulation of sterol regulatory element-binding protein-1c in mice: a novel peroxisome proliferator-activated receptor alpha-independent mechanism. AB - The triglyceride-lowering effect of bezafibrate in humans has been attributed to peroxisome proliferator-activated receptor (PPAR) alpha activation based on results from rodent studies. However, the bezafibrate dosages used in conventional rodent experiments are typically higher than those in clinical use (> or =50 versus < or =10 mg/kg/day), and thus it remains unclear whether such data can be translated to humans. Furthermore, because bezafibrate is a pan-PPAR activator, the actual contribution of PPARalpha to its triglyceride-lowering properties remains undetermined. To address these issues, bezafibrate at clinically relevant doses (10 mg/kg/day; low) was administered to wild-type and Ppara-null mice, and its effects were compared with those from conventionally used doses (100 mg/kg/day; high). Pharmacokinetic analyses showed that maximum plasma concentration and area under the concentration-time curve in bezafibrate treated mice were similar to those in humans at low doses, but not at high doses. Low-dose bezafibrate decreased serum/liver triglycerides in a PPARalpha independent manner by attenuation of hepatic lipogenesis and triglyceride secretion. It is noteworthy that instead of PPAR activation, down-regulation of sterol regulatory element-binding protein (SREBP)-1c was observed in mice undergoing low-dose treatment. High-dose bezafibrate decreased serum/liver triglycerides by enhancement of hepatic fatty acid uptake and beta-oxidation via PPARalpha activation, as expected. In conclusion, clinically relevant doses of bezafibrate exert a triglyceride-lowering effect by suppression of the SREBP-1c regulated pathway in mice and not by PPARalpha activation. Our results may provide novel information about the pharmacological mechanism of bezafibrate action and new insights into the treatment of disorders involving SREBP-1c. PMID- 19124613 TI - Alanine scanning mutagenesis of the second extracellular loop of type 1 corticotropin-releasing factor receptor revealed residues critical for peptide binding. AB - Upon binding of the corticotropin-releasing factor (CRF) analog sauvagine to the type 1 CRF receptor (CRF(1)), the amino-terminal portion of the peptide has been shown to lie near Lys257 in the receptor's second extracellular loop (EL2). To test the hypothesis that EL2 residues play a role in the binding of sauvagine to CRF(1) we carried out an alanine-scanning mutagenesis study to determine the functional role of EL2 residues (Leu251 to Val266). Only the W259A, F260A, and W259A/F260A mutations reduced the binding affinity and potency of sauvagine. In contrast, these mutations did not seem to significantly alter the overall receptor conformation, in that they left unchanged the affinities of the ligands astressin and antalarmin that have been suggested to bind to different regions of CRF(1). The W259A, F260A, and W259A/F260A mutations also decreased the affinity of the endogenous ligand, CRF, implying that these residues may play a common important role in the binding of different peptides belonging to CRF family. Parallel amino acid deletions of the two peptides produced ligands with various affinities for wild-type CRF(1) compared with the W259A, F260A, and W259A/F260A mutants, supporting the interaction between the amino-terminal residues 8 to 10 of sauvagine and the corresponding region in CRF with EL2 of CRF(1). This is the first time that a specific region of CRF(1) has been implicated in detailed interactions between the receptor and the amino-terminal portion of peptides belonging to the CRF family. PMID- 19124614 TI - Functional somatic symptoms and psychological states: an electronic diary study. AB - OBJECTIVE: To investigate the links between functional physical symptoms and psychological states in a sample of patients with persistent medically unexplained symptoms. Despite the epidemiological evidence for links between physical symptoms and mental processes, prior diary studies have shown inconsistent associations and generally been limited to single symptom and psychological variable pairs. METHODS: Twenty-six patients with at least three functional physical symptoms completed twice daily self-report measures of symptoms, fatigue, anxiety, stress, mood, and symptom concern using electronic diaries over 12 weeks. Associations between physical symptoms and psychological variables were measured by linear mixed effects models at the levels of diary entry and individual. RESULTS: Despite high baseline questionnaire scores for depression and anxiety, diary ratings of anxiety and stress were relatively low. Fixed effects regression coefficients varied between symptoms and psychological variables; for instance, the fixed effects regression coefficient (95% Confidence Intervals) for fatigue as the outcome variable was 0.39 (0.31-0.47) with low mood and 0.05 (-0.01-0.10) with stress as the predictor. Random effects coefficients showed less variation between individuals for fatigue and musculoskeletal pain than for other symptoms. CONCLUSION: Self-reported mood and symptom concern were more strongly associated with functional physical symptoms than anxiety or stress. We suggest that one reason patients with functional somatic symptoms reject psychosomatic explanations is because they do not experience sufficient correlation between symptoms and psychological states. PMID- 19124615 TI - Repolarization changes induced by mental stress in normal subjects and patients with coronary artery disease: effect of nitroglycerine. AB - OBJECTIVES: Mental stress can significantly affect ventricular repolarization, which could potentially trigger arrhythmias. We compared the effect of mental stress on repolarization indexed by the amplitude and area of the T wave in patients with coronary artery disease (CAD) and healthy subjects. METHODS: Fourteen healthy controls (11 M, mean age 42 years) and 14 patients with stable CAD (12 M, mean age 64) underwent a mental stress protocol consisting of mental arithmetic followed by a speech (5 minutes each), which was performed on two occasions following either nitroglycerine (NTG) or placebo. Multiple 12-lead electrocardiograms were acquired and repolarization was analyzed using automatically measured T wave amplitude (T(amp)) and area (T(area)). RESULTS: When preceded by placebo the overall effect of mental stress, whether induced by arithmetic or speech, was significantly different in CAD patients compared with controls, with a decrease in T(amp) and T(area) in controls and an increase in patients; e.g., change in T(amp) during arithmetic -20 +/- 3 microV in controls versus 4 +/- 2 microV in patients, p < .001, and during speech -9 +/- 3 microV in controls versus 7 +/- 1 microV in patients, p < .001. Following NTG, the effect of stress on repolarization was similar in the 2 groups, with a reversed effect, i.e., decrease instead of increase in T(amp) and T(area) in CAD patients. CONCLUSIONS: The effect of mental stress on ventricular repolarization is significantly different in CAD patients compared with healthy controls. These differences are considerably reduced by NTG. PMID- 19124616 TI - Lifetime cognitive performance is associated with midlife physical performance in a prospective national birth cohort study. AB - OBJECTIVES: To examine whether measures of cognitive performance across life are related to physical performance at age 53 years, allowing for potential confounders. METHODS: In a large representative British birth cohort of men and women (N = 2135) the associations between cognitive performance across life (i.e., standardized cognition scores at ages 15, 43, and 53 years and changes in verbal memory and search speed scores between 43 and 53 years) and measures of physical performance at age 53 years (i.e., standing balance, chair rising, and grip strength) were examined. Adjustments were made for body size, physical activity levels, health status, and socioeconomic conditions at age 53 years. RESULTS: Higher cognitive scores on all childhood and adult tests, and a slower decline in verbal memory and search speed, were associated with better standing balance. Higher verbal fluency scores and a slower decline in verbal memory and search speed were more strongly related than scores on tests of general cognitive ability to chair rising. The relationships between cognitive performance and grip strength were inconsistent and weak. CONCLUSIONS: The differential patterns of association found are consistent with the degree to which each is dependent on central nervous system function. Our findings suggest that initial developmental differences as well as shared ageing processes may underlie associations found between cognitive and physical performance. PMID- 19124617 TI - A noninvasive clinically useful predictor for mental stress-induced ischemia. PMID- 19124618 TI - What's in a name? Psychosomatic medicine and biobehavioral medicine. PMID- 19124619 TI - Purim CPR. PMID- 19124620 TI - When husband and wife become doctor and patient: a lesson for modern health care. PMID- 19124621 TI - Exploring exposure to mobile-phone electromagnetic fields and psychophysiological and self-rated symptoms. PMID- 19124622 TI - Coping with genetic testing for breast cancer susceptibility. AB - OBJECTIVE: To examine how women cope with genetic testing for heightened susceptibility to breast cancer. METHODS: Participants were 126 White women (age = 44 +/- 9 years) who were participants in a larger study of genetic testing for risk of different chronic diseases. All women were at higher-than-average risk for breast cancer due to a personal and/or family history and were considering genetic testing. Distress (Symptom Checklist-90-Revised, Impact of Event Scale, Perceived Stress Scale, Spielberger State-Trait Anxiety Inventory, and the Center for Epidemiological Studies Depression Scale) was assessed at four assessments; one before and three after the decision to have genetic testing. The majority of women (n = 100) had testing. The follow-up assessments occurred at 1 week after receiving results (or 3-4 months after baseline if testing was not elected), and then at 3 and 6 months after the second assessment. Coping (Brief COPE) was measured at the first and third assessments. RESULTS: Coping was relatively stable over time and did not vary as a function of genetic test results. Active coping strategies were used more often by women with a personal cancer history than by women without cancer. Use of avoidant coping was reliably and positively associated with distress over time independent of cancer history and test result. CONCLUSIONS: The identification of specific coping styles that were associated with more or less distress is useful as a means of identifying and targeting coping interventions and predicting which participants may be at risk for distress. PMID- 19124623 TI - Changes of body composition in bulimia nervosa: increased visceral fat and adrenal gland size. AB - OBJECTIVE: To study the hypothesis that young women suffering from active bulimia nervosa (BN) have more visceral fat and increased adrenal gland volumes (AGV) than healthy controls (HC). METHODS: Thirteen patients with BN of purging type and 11 healthy age and weight matched women (HC), aged between 19 and 36 years (mean 24 +/- 3 years), with a BMI of 19 to 29 (mean 24 +/- SD 3) were examined. BN was diagnosed by DSM-IV criteria and the severity of illness by the Eating Disorder Inventory (EDI-2). Whole body fat distribution and AGV were determined using a whole body magnetic resonance (MR) scan (T1w) and a 3D-sequence (T1w) at 1.5 Tesla. Salivary cortisol was determined at 9 AM and 4 PM. RESULTS: BN patients had significantly more visceral adipose tissue (VAT) (HC, 1589.3 +/- 967.6 ml versus 927.2 +/- 428.4 ml, p < .05) and an increased relative AGV (0.068% of body volume versus 0.048% of body volume, p < .05) compared with HC, although waist circumference and BMI did not differ. Although the VAT part in the upper abdomen was increased, especially the VAT of lower abdomen along with the pelvis or any subcutaneous fat compartment was not increased. CONCLUSIONS: The increase of the VAT volume and the increased AGV in BN women with purging point to chronic high stress levels associated with a hyperactivity of the hypothalamic pituitary-adrenal axis in these patients. This is the first MR study showing morphological changes in stress associated endocrine organs of young BN patients. PMID- 19124624 TI - Independent association between lower level of social support and higher coagulation activity before and after acute psychosocial stress. AB - OBJECTIVE: To investigate the relationship between social support and coagulation parameter reactivity to mental stress in men and to determine if norepinephrine is involved. Lower social support is associated with higher basal coagulation activity and greater norepinephrine stress reactivity, which in turn, is linked with hypercoagulability. However, it is not known if low social support interacts with stress to further increase coagulation reactivity or if norepinephrine affects this association. These findings may be important for determining if low social support influences thrombosis and possible acute coronary events in response to acute stress. We investigated the relationship between social support and coagulation parameter reactivity to mental stress in men and determined if norepinephrine is involved. METHODS: We measured perceived social support in 63 medication-free nonsmoking men (age (mean +/- standard error of the mean) = 36.7 +/- 1.7 years) who underwent an acute standardized psychosocial stress task combining public speaking and mental arithmetic in front of an audience. We measured plasma D-dimer, fibrinogen, clotting Factor VII activity (FVII:C), and plasma norepinephrine at rest as well as immediately after stress and 20 minutes after stress. RESULTS: Independent of body mass index, mean arterial pressure, and age, lower social support was associated with higher D-dimer and fibrinogen levels at baseline (p < .012) and with greater increases in fibrinogen (beta = 0.36, p = .001; DeltaR(2) = .12), and D-dimer (beta = -0.21, p = .017; DeltaR(2) = .04), but not in FVII:C (p = .83) from baseline to 20 minutes after stress. General linear models revealed significant main effects of social support and stress on fibrinogen, D-dimer, and norepinephrine (p < .035). Controlling for norepinephrine did not change the significance of the reported associations between social support and the coagulation measures D-dimer and fibrinogen. CONCLUSIONS: Our results suggest that lower social support is associated with greater coagulation activity before and after acute stress, which was unrelated to norepinephrine reactivity. PMID- 19124625 TI - Optimism: a good theme for family medicine. PMID- 19124627 TI - Retaining optimism in the face of adversity. PMID- 19124628 TI - Effectiveness of vitamin B12 in treating recurrent aphthous stomatitis: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: The frequency of recurrent aphthous stomatitis (RAS), the most common oral mucosa lesions seen in primary care, is up to 25% in the general population. However, there has been no optimal therapeutic approach. Our objective was to confirm our previous clinical observation of the beneficial treatment of RAS with vitamin B(12). METHODS: A randomized, double-blind, placebo-controlled trial was done using primary care patients. A sublingual a dose of 1000 mcg of vitamin B(12) was used in patients in the intervention group for 6 months. RESULTS: In total, 58 patients suffering from RAS participated in the study: 31 were included in the intervention group and 27 were included in control group. All parameters of RAS among patients in the intervention group were recorded and compared with the control group. The duration of outbreaks, the number of ulcers, and the level of pain were reduced significantly (P < .05) at 5 and 6 months of treatment with vitamin B(12), regardless of initial vitamin B(12) levels in the blood. During the last month of treatment a significant number of participants in the intervention group reached "no aphthous ulcers status" (74.1% vs 32.0%; P < .01). CONCLUSION: Vitamin B(12) treatment, which is simple, inexpensive, and low-risk, seems to be effective for patients suffering from RAS, regardless of the serum vitamin B(12) level. PMID- 19124629 TI - Hear today, gone tomorrow: an assessment of portable entertainment player use and hearing acuity in a community sample. AB - BACKGROUND: Noise-induced hearing loss (NIHL) is a common but preventable disability. The purpose of this study was to assess the understanding of NIHL in a community sample in the context of exposure to portable music players, including MP3 players, and personal hearing acuity as tested with the Welch Allyn Audioscope 3. METHODS: A cross-sectional convenience sample of 94 adults (18 to 65 years old) at a university recreation center completed an analysis of personal use of portable digital music players (MP3 players), concerns about hearing loss, and a 3-dB-level hearing test at 4 levels of speech frequency in a low ambient noise setting. RESULTS: The majority of participants (85%) were concerned about hearing loss, willing to protect their hearing with lower volume (77%), had little measurable hearing loss but were exposed to longer and louder periods of noise than other national samples, and mistakenly felt that NIHL is a medically reversible condition. Many (40%) also wanted their family medicine physician to be more concerned about their hearing. CONCLUSIONS: Family medicine physicians are in a key position to provide basic information on the preventability and negative consequences of NIHL, as well as to identify and refer patients with identified hearing loss. PMID- 19124630 TI - Perceived complexity of care, perceived autonomy, and career satisfaction among primary care physicians. AB - BACKGROUND: The purpose of this study was to examine relationships of both perceived autonomy and perceived complexity of care with career satisfaction. METHODS: This secondary analysis used 3 consecutive surveys of family physicians, internists, and pediatricians from the Community Tracking Survey. Two-way analysis of variance assessed interaction effects of perceived complexity of care and perceived autonomy on satisfaction. Logistic regression analysis identified physician characteristics, practice characteristics, practice improvement strategies, perceived complexity, and perceived autonomy that accounted for variance in career satisfaction among physicians. RESULTS: Although 24% to 27% of physicians felt perceived complexity of care expected was greater than it should be, 83% to 86% felt free to make clinical decisions. Approximately 80% of physicians were satisfied with their careers. Differences in probability of career satisfaction were highly significant (P < .001) for both perceived complexity of care and perceived autonomy as well as their interaction. A multiphysician practice; the ability to obtain high quality ancillary services (such as physical therapy, home health care, and nutritional counseling); managed care revenue, lower levels of perceived complexity of expected care; and perceived autonomy were consistently associated with satisfaction. CONCLUSION: Higher perceived autonomy and lower perceived patient complexity as higher than desirable were associated with high career satisfaction among primary care physicians. PMID- 19124632 TI - Women at high risk for breast cancer--what the primary care provider needs to know. AB - Until recently, the assessment made by the primary care provider regarding screening for breast cancer was generally limited to decisions about when to initiate mammography. Early diagnosis was stressed as the best protection against breast cancer morbidity. However, there have been recent developments in the ability to predict and modify breast cancer risk. It is therefore important for the primary care provider to be able to identify women at higher risk for breast cancer and be familiar with issues regarding screening and risk reduction. Recent data regarding the evaluation of breast cancer risk, newer screening strategies for high-risk women, and medical and surgical approaches to reduce breast cancer risk and are discussed in this article. PMID- 19124631 TI - Variables associated with human papillomavirus (HPV) vaccine acceptance by men. AB - OBJECTIVE: To determine correlates of human papillomavirus (HPV) vaccine acceptance for men. METHODS: A convenience sample of men aged 18 to 45 years read a one-page information sheet about HPV and the HPV vaccine, then completed a 29 item questionnaire. chi(2) tests were used to determine whether differences in demographic, sexual, and vaccine-related variables existed between levels of wanting the HPV vaccine. RESULTS: Positive correlates of HPV vaccine acceptance included higher education (P < .0001); hispanic ethnicity (P = .0003); wearing a seat belt most of the time (P = .02); regular tobacco use (P = <.001); not being sexually active (P = .0008); history of more than 10 female sexual partners (P = .0004); not having oral sex (P = .045); extreme worry about vaccine side effects (P < .0001); extreme concern about vaccine safety (P < .0001); the importance of getting vaccines (P < .0001); familiarity with HPV (P < .0001); and extreme importance of receiving the HPV vaccine (P < .0001). CONCLUSIONS: Men with greater general education, high-risk behaviors, and knowledge about HPV are more likely to want the HPV vaccine. Focused educational efforts should facilitate even wider acceptance of the HPV vaccine by men. PMID- 19124633 TI - An appraisal of dual antiplatelet therapy with clopidogrel and aspirin for prevention of cardiovascular events. AB - Combination antiplatelet therapy, typically with clopidogrel and aspirin, is commonly used for the prevention of cardiovascular events. When used for appropriate indications and duration, its benefits clearly outweigh its risks. However, it is not uncommon for the combination to be used outside of recommended indications or for longer than recommended durations. In these circumstances data are at best unclear and, at worst, indicative of harm. Furthermore, use for one of its indications-prevention of cardiac events after deployment of a coronary stent-is complicated by the type of stent used. This report reviews the evidence surrounding combination antiplatelet therapy with clopidogrel and aspirin, with an emphasis on identifying appropriate indications for and durations of therapy. PMID- 19124634 TI - Thirty years of family medicine publications in Israel (1975-2004): what, where, and how much? AB - BACKGROUND: Departments of family medicine in Israel were established in the 1970s. Until now, little or no effort has been made to characterize the productivity of Israeli board-certified family medicine physicians in publishing peer-reviewed scientific articles. METHODS: Publications were identified by 2 methods. First, a PubMed search by names of current and past faculty from all the departments of family medicine in Israel (1975-2004). Secondly, all of the departments in Israel forwarded a list of all publications by their faculty (including those that do not appear in Medline). The abstracts of all publications were extracted and were separately and blindly evaluated by 2 reviewers. Publications were classified according to Medline citation, language, journal impact factor, and publication type. RESULTS: A total of 1165 publications were identified and analyzed. More than half of the articles were published in the last 10 years. Seventy-two percent were cited in Medline. Publications in English encompassed 64.7% of the publications, Hebrew 34.6%, and 0.7% other. Approximately 6% of the articles were published in journals with impact factor > or =3, with research articles accounting for 46.9%. The publication output of family medicine in Israel averaged 85.4 publications per 1000 family medicine physicians per year. Almost 70% of the articles were published in non-family medicine journals. Academically affiliated, board certified family medicine physicians published at higher rates, averaging 334.3 per 1000 academic family medicine physicians per year (data available for 2000 2004 only). CONCLUSIONS: Publishing and research are important to the development of family medicine as an academic profession, in which Israeli family physicians show significant productivity. PMID- 19124636 TI - Improvement of chronic back pain or failed back surgery with vitamin D repletion: a case series. AB - This article reviews 6 selected cases of improvement/resolution of chronic back pain or failed back surgery after vitamin D repletion in a Canadian family practice setting. Pub Med was searched for articles on chronic back pain, failed back surgery, and vitamin D deficiency. Chronic low back pain and failed back surgery may improve with repletion of vitamin D from a state of deficiency/insufficiency to sufficiency. Vitamin D insufficiency is common; repletion of vitamin D to normal levels in patients who have chronic low back pain or have had failed back surgery may improve quality of life or, in some cases, result in complete resolution of symptoms. PMID- 19124635 TI - Overtreating chronic back pain: time to back off? AB - Chronic back pain is among the most common patient complaints. Its prevalence and impact have spawned a rapidly expanding range of tests and treatments. Some of these have become widely used for indications that are not well validated, leading to uncertainty about efficacy and safety, increasing complication rates, and marketing abuses. Recent studies document a 629% increase in Medicare expenditures for epidural steroid injections; a 423% increase in expenditures for opioids for back pain; a 307% increase in the number of lumbar magnetic resonance images among Medicare beneficiaries; and a 220% increase in spinal fusion surgery rates. The limited studies available suggest that these increases have not been accompanied by population-level improvements in patient outcomes or disability rates. We suggest a need for a better understanding of the basic science of pain mechanisms, more rigorous and independent trials of many treatments, a stronger regulatory stance toward approval and post-marketing surveillance of new drugs and devices for chronic pain, and a chronic disease model for managing chronic back pain. PMID- 19124637 TI - Kummel disease: a not-so-rare complication of osteoporotic vertebral compression fractures. AB - Kummel disease is the eponym for avascular necrosis of the vertebral body after a vertebral compression fracture. As our population ages, the prevalence of osteoporosis, its most common fragility fracture (vertebral compression fracture), and Kummel disease will increase. The purpose of this article is to report a prototypical case with complete radiographic and histologic workup and to provide facts regarding Kummel disease that are salient to the primary care provider. PMID- 19124638 TI - Lemierre syndrome following oropharyngeal infection: a case series. AB - Lemierre syndrome is a potentially fatal complication of oropharyngeal infections that is characterized by jugular vein thrombosis and septic emboli. Fusobacterium spp. are the causative organisms. Patients typically present with high fevers, neck pain, and pulmonary symptoms. Prompt clinical diagnosis and treatment with appropriate antibiotics are critical. Anticoagulation for this syndrome is controversial. We present 3 cases within a 12-month period of young, family practice patients seen with oropharyngeal infections who developed Lemierre syndrome. The incidence of Lemierre syndrome may be increasing and primary care physicians should be aware of this rare though serious condition. PMID- 19124639 TI - Retropharyngeal calcific tendonitis: report of two cases. AB - Retropharyngeal calcific tendonitis is an inflammatory process of the superior oblique tendons of the longus colli muscle, a neck flexor in the upper cervical spine, caused by deposition of calcium hydroxyapatite crystals; the definitive diagnostic test is computed tomography (CT). Presented in this article are two cases seen at our institution. Patients typically present with acute onset of neck pain/spasm, odynophagia, dysphagia, and/or low grade fevers. Leukocytosis and elevated erythrocyte sedimentation rate may be noted. It is important to understand this entity because its signs and symptoms are mimickers of those of the more serious condition of retropharyngeal space abscess. Calcific tendonitis is managed conservatively whereas retropharyngeal abscess requires incision and drainage. Some may argue that this entity is a zebra because its reported incidence in the literature is low. However, most of these studies were done in an era when CT was not yet in vogue. With today's widespread use of CT and its superb ability to visualize the calcification, the true incidence of this condition is probably higher and, thus, it is important for the family practitioner to be aware of this entity. The astute clinician may save the patient from unnecessary diagnostic workup, undue anxiety, and delays in hospital discharge. PMID- 19124640 TI - Bullous lesions on a skin graft donor site. PMID- 19124641 TI - Re: the patient-centered medical home movement--promise and peril for family medicine. PMID- 19124642 TI - Re: outcomes from treatment of infertility with natural procreative technology in an Irish general practice. PMID- 19124644 TI - NARP syndrome in a patient harbouring an insertion in the MT-ATP6 gene that results in a truncated protein. AB - BACKGROUND: Neurogenic muscle weakness, ataxia and retinitis pigmentosa (NARP) syndrome have been associated to m.8993T>G/C mutations in the subunit 6 of the ATP synthase (p.MT-ATP6). METHODS: We have performed a mutational screening of the mitochondrial DNA gene encoding for this protein in 62 patients with the disease, that do not carry any of the common mutations described to date. RESULTS: We report clinical and molecular data in one patient who harbours a de novo insertion in the MT-ATP6 gene that results in a truncated subunit. The mutation was heteroplasmic (85%) in muscle DNA and the BN-PAGE analysis showed a clear decrease in the amount of ATP synthase. CONCLUSION: Molecular analysis of NARP patients cannot be limited to the search of the m.8993T>G/C and either the ATP6 or the whole mtDNA should be sequenced. PMID- 19124645 TI - Familial occurrence of schwannomas and malignant rhabdoid tumour associated with a duplication in SMARCB1. AB - BACKGROUND: The role of germline and somatic SMARCB1 gene mutations in malignant rhabdoid tumour (MRT) predisposition is well known. Germline SMARCB1 mutations have also recently been identified in a subset of individuals with schwannomatosis. Surprisingly, MRT predisposition and schwannomatosis have never been reported to co-occur in a family. The correlation between genotype and phenotype for mutations in SMARCB1 has not been determined. RESULTS: We have identified a germline 2631 bp duplication that includes exon 6 of SMARCB1 in a unique family with a four generation history of MRT predisposition and schwannomatosis. This duplication segregates with disease in individuals affected with both conditions, linking MRT predisposition and schwannomatosis as components of the same syndrome in this family. CONCLUSION: The unique combination of tumours that result from the duplication described in this report may provide important clues about the mechanisms that influence the phenotype associated with a given SMARCB1 mutation. PMID- 19124647 TI - Can clinicians be scientists? PMID- 19124646 TI - Thieno[2,3-c]isoquinolin-5-one, a potent poly(ADP-ribose) polymerase inhibitor, promotes atherosclerotic plaque regression in high-fat diet-fed apolipoprotein E deficient mice: effects on inflammatory markers and lipid content. AB - We recently showed that poly(ADP-ribose) polymerase (PARP) is activated within atherosclerotic plaques in an animal model of atherosclerosis. Pharmacological inhibition of PARP or reduced expression in heterozygous animals interferes with atherogenesis and may promote factors of plaque stability, possibly reflecting changes in inflammatory and cellular factors consistent with plaque stability. The current study addresses the hypothesis that pharmacological inhibition of PARP promotes atherosclerotic plaque regression. Using a high-fat diet-induced atherosclerosis apolipoprotein E(-/-) mouse model, we demonstrate that administration of the potent PARP inhibitor, thieno[2,3-c]isoquinolin-5-one (TIQ A), when combined with a regular diet regimen during treatment, induced regression of established plaques. Plaque regression was associated with a reduction in total cholesterol and low-density lipoproteins. Furthermore, plaques of TIQ-A-treated mice were highly enriched with collagen and smooth muscle cells, displayed thick fibrous caps, and exhibited a marked reduction in CD68-positive macrophage recruitment and associated foam cell presence. These changes correlated with a significant decrease in expression of monocyte chemoattractant protein-1 and intercellular cell adhesion molecule-1, potentially as a result of a robust reduction in tumor necrosis factor expression. The PARP inhibitor appeared to affect cholesterol metabolism by affecting acyl-coenzymeA/cholesterol acyltransferase-1 expression but exerted no effect on cholesterol influx or efflux as assessed by an examination of the ATP-binding cassette transporter-1 and the scavenger receptor-A expression levels in the different experimental groups. In accordance, PARP inhibition may prove beneficial not only in preventing atherogenesis but also in promoting regression of preexisting plaques. PMID- 19124648 TI - Between-word simplification patterns in the continuous speech of children with speech sound disorders. AB - PURPOSE: This study was designed to identify and describe between-word simplification patterns in the continuous speech of children with speech sound disorders. It was hypothesized that word combinations would reveal phonological changes that were unobserved with single words, possibly accounting for discrepancies between the intelligibility of single-word samples and that of continuous speech. METHOD: Four boys with developmental speech sound disorders provided samples of single words and continuous speech. Substitutions and deletions with single words formed the basis for determining 2 categories of between-word segment mismatches: observed and novel. Mismatches were attributed to one of 4 types of between-word simplifications reported for typical phonological development: between-word consonant deletion, between-word cluster reduction, between-word consonant sequence reduction, and between-word assimilation. RESULTS: Continuous speech revealed observed and novel patterns. Segment mismatches occurred differentially among potential between-word simplification environments. The most frequently occurring novel pattern involved the deletion of a coda consonant within a between-word consonant sequence. CONCLUSIONS: Children with speech sound disorders demonstrated substitutions and deletions between words in continuous speech that may not be predicted on the basis of single-word productions. The identification of potential contexts for such mismatches may serve as a framework for the assessment of continuous speech samples of children with speech sound disorders in health care and school settings. PMID- 19124649 TI - The accuracy of parent and teacher reports in assessing the vocabulary knowledge of Chinese children with hearing impairment. AB - PURPOSE: This study aimed to investigate (a) the accuracy of adult reports in assessing the vocabulary knowledge of Cantonese-speaking children with hearing impairment (HI) and (b) the factors that are associated with the accuracy of those reports. METHOD: The first participant group consisted of 47 children and their mothers. The second group consisted of 47 children and 21 teachers. All of the children had profound HI, with a mean age of 57 months. The ratings of the adults were compared with the children's test scores in a standardized test. RESULTS: Both adult groups reported children's performance with satisfactory accuracy. Mother and teacher ratings showed fair to good agreement with each other. Two factors significantly associated with the accuracy of the report were (a) the difficulty of the vocabulary and (b) the child's vocabulary ability as determined by the standardized norm-referenced receptive vocabulary test. The mothers' education level, occupation, and socioeconomic status, and the teachers' teaching experience and length of time they had trained the child, were not significant factors. CONCLUSION: The results suggest that employing adult reports to collect data on the word knowledge of children with HI is applicable to a broad spectrum of the Cantonese-speaking Chinese population. PMID- 19124650 TI - Accelerating preschoolers' early literacy development through classroom-based teacher-child storybook reading and explicit print referencing. AB - PURPOSE: This study examined the impact of teacher use of a print referencing style during classroom-based storybook reading sessions conducted over an academic year. Impacts on preschoolers' early literacy development were examined, focusing specifically on the domain of print knowledge. METHOD: This randomized, controlled trial examined the effects of a print referencing style on 106 preschool children attending 23 classrooms serving disadvantaged preschoolers. Following random assignment, teachers in 14 classrooms used a print referencing style during 120 large-group storybook reading sessions during a 30-week period. Teachers in 9 comparison classrooms read at the same frequency and with the same storybooks but used their normal style of reading. RESULTS: Children whose teachers used a print referencing style showed larger gains on 3 standardized measures of print knowledge: print concept knowledge, alphabet knowledge, and name writing, with medium-sized effects. CLINICAL IMPLICATIONS: The convergence of the present findings with those of previous efficacy studies indicates that print referencing intervention can be used confidently as an approach for facilitating print knowledge in preschool-age children. Speech-language pathologists can serve an important role in supporting preschool educators as they use this evidence-based technique with pupils in their classrooms. PMID- 19124651 TI - Progenitor cell proliferation in the retina is dependent on Notch-independent Sonic hedgehog/Hes1 activity. AB - Sonic hedgehog (Shh) is an indispensable, extrinsic cue that regulates progenitor and stem cell behavior in the developing and adult mammalian central nervous system. Here, we investigate the link between the Shh signaling pathway and Hes1, a classical Notch target. We show that Shh-driven stabilization of Hes1 is independent of Notch signaling and requires the Shh effector Gli2. We identify Gli2 as a primary mediator of this response by showing that Gli2 is required for Hh (Hedgehog)-dependent up-regulation of Hes1. We also show using chromatin immunoprecipitation that Gli2 binds to the Hes1 promoter, which suggests that Hes1 is a Hh-dependent direct target of Gli2 signaling. Finally, we show that Shh stimulation of progenitor proliferation and cell diversification requires Gli2 and Hes1 activity. This paper is the first demonstration of the mechanistic and functional link between Shh, Gli, and Hes1 in the regulation of progenitor cell behavior. PMID- 19124652 TI - USP7 counteracts SCFbetaTrCP- but not APCCdh1-mediated proteolysis of Claspin. AB - Claspin is an adaptor protein that facilitates the ataxia telangiectasia and Rad3 related (ATR)-mediated phosphorylation and activation of Chk1, a key effector kinase in the DNA damage response. Efficient termination of Chk1 signaling in mitosis and during checkpoint recovery requires SCF(betaTrCP)-dependent destruction of Claspin. Here, we identify the deubiquitylating enzyme ubiquitin specific protease 7 (USP7) as a novel regulator of Claspin stability. Claspin and USP7 interact in vivo, and USP7 is required to maintain steady-state levels of Claspin. Furthermore, USP7-mediated deubiquitylation markedly prolongs the half life of Claspin, which in turn increases the magnitude and duration of Chk1 phosphorylation in response to genotoxic stress. Finally, we find that in addition to the M phase-specific, SCF(betaTrCP)-mediated degradation, Claspin is destabilized by the anaphase-promoting complex (APC) and thus remains unstable in G1. Importantly, we demonstrate that USP7 specifically opposes the SCF(betaTrCP)- but not APC(Cdh1)-mediated degradation of Claspin. Thus, Claspin turnover is controlled by multiple ubiquitylation and deubiquitylation activities, which together provide a flexible means to regulate the ATR-Chk1 pathway. PMID- 19124653 TI - Htm1 protein generates the N-glycan signal for glycoprotein degradation in the endoplasmic reticulum. AB - To maintain protein homeostasis in secretory compartments, eukaryotic cells harbor a quality control system that monitors protein folding and protein complex assembly in the endoplasmic reticulum (ER). Proteins that do not fold properly or integrate into cognate complexes are degraded by ER-associated degradation (ERAD) involving retrotranslocation to the cytoplasm and proteasomal peptide hydrolysis. N-linked glycans are essential in glycoprotein ERAD; the covalent oligosaccharide structure is used as a signal to display the folding status of the host protein. In this study, we define the function of the Htm1 protein as an alpha1,2-specific exomannosidase that generates the Man(7)GlcNAc(2) oligosaccharide with a terminal alpha1,6-linked mannosyl residue on degradation substrates. This oligosaccharide signal is decoded by the ER-localized lectin Yos9p that in conjunction with Hrd3p triggers the ubiquitin-proteasome-dependent hydrolysis of these glycoproteins. The Htm1p exomannosidase activity requires processing of the N-glycan by glucosidase I, glucosidase II, and mannosidase I, resulting in a sequential order of specific N-glycan structures that reflect the folding status of the glycoprotein. PMID- 19124654 TI - Lamin A/C-mediated neuromuscular junction defects in Emery-Dreifuss muscular dystrophy. AB - The LMNA gene encodes lamins A and C, two intermediate filament-type proteins that are important determinants of interphase nuclear architecture. Mutations in LMNA lead to a wide spectrum of human diseases including autosomal dominant Emery Dreifuss muscular dystrophy (AD-EDMD), which affects skeletal and cardiac muscle. The cellular mechanisms by which mutations in LMNA cause disease have been elusive. Here, we demonstrate that defects in neuromuscular junctions (NMJs) are part of the disease mechanism in AD-EDMD. Two AD-EDMD mouse models show innervation defects including misexpression of electrical activity-dependent genes and altered epigenetic chromatin modifications. Synaptic nuclei are not properly recruited to the NMJ because of mislocalization of nuclear envelope components. AD-EDMD patients with LMNA mutations show the same cellular defects as the AD-EDMD mouse models. These results suggest that lamin A/C-mediated NMJ defects contribute to the AD-EDMD disease phenotype and provide insights into the cellular and molecular mechanisms for the muscle-specific phenotype of AD-EDMD. PMID- 19124655 TI - Rcan1 negatively regulates Fc epsilonRI-mediated signaling and mast cell function. AB - Aggregation of the high affinity IgE receptor (Fc epsilonRI) activates a cascade of signaling events leading to mast cell activation. Subsequently, inhibitory signals are engaged for turning off activating signals. We identified that regulator of calcineurin (Rcan) 1 serves as a negative regulator for turning off Fc epsilonRI-mediated mast cell activation. Fc epsilonRI-induced Rcan1 expression was identified by suppression subtractive hybridization and verified by real-time quantitative polymerase chain reaction and Western blotting. Deficiency of Rcan1 led to increased calcineurin activity, increased nuclear factor of activated T cells and nuclear factor kappaB activation, increased cytokine production, and enhanced immunoglobulin E-mediated late-phase cutaneous reactions. Forced expression of Rcan1 in wild-type or Rcan1-deficient mast cells reduced Fc epsilonRI-mediated cytokine production. Rcan1 deficiency also led to increased Fc epsilonRI-mediated mast cell degranulation and enhanced passive cutaneous anaphylaxis. Analysis of the Rcan1 promoter identified a functional Egr1 binding site. Biochemical and genetic evidence suggested that Egr1 controls Rcan1 expression. Our results identified Rcan1 as a novel inhibitory signal in Fc epsilonRI-induced mast cell activation and established a new link of Egr1 and Rcan1 in Fc epsilonRI signaling. PMID- 19124656 TI - Down-regulation of CYLD expression by Snail promotes tumor progression in malignant melanoma. AB - High malignancy and early metastasis are hallmarks of melanoma. Here, we report that the transcription factor Snail1 inhibits expression of the tumor suppressor CYLD in melanoma. As a direct consequence of CYLD repression, the protooncogene BCL-3 translocates into the nucleus and activates Cyclin D1 and N-cadherin promoters, resulting in proliferation and invasion of melanoma cells. Rescue of CYLD expression in melanoma cells reduced proliferation and invasion in vitro and tumor growth and metastasis in vivo. Analysis of a tissue microarray with primary melanomas from patients revealed an inverse correlation of Snail1 induction and loss of CYLD expression. Importantly, tumor thickness and progression-free and overall survival inversely correlated with CYLD expression. Our data suggest that Snail1-mediated suppression of CYLD plays a key role in melanoma malignancy. PMID- 19124657 TI - Efferocytosis impairs pulmonary macrophage and lung antibacterial function via PGE2/EP2 signaling. AB - The ingestion of apoptotic cells (ACs; termed "efferocytosis") by phagocytes has been shown to trigger the release of molecules such as transforming growth factor beta, interleukin-10 (IL-10), nitric oxide, and prostaglandin E(2) (PGE(2)). Although the antiinflammatory actions of these mediators may contribute to the restoration of homeostasis after tissue injury, their potential impact on antibacterial defense is unknown. The lung is highly susceptible to diverse forms of injury, and secondary bacterial infections after injury are of enormous clinical importance. We show that ACs suppress in vitro phagocytosis and bacterial killing by alveolar macrophages and that this is mediated by a cyclooxygenase-PGE(2)-E prostanoid receptor 2 (EP2)-adenylyl cyclase-cyclic AMP pathway. Moreover, intrapulmonary administration of ACs demonstrated that PGE(2) generated during efferocytosis and acting via EP2 accounts for subsequent impairment of lung recruitment of polymorphonuclear leukocytes and clearance of Streptococcus pneumoniae, as well as enhanced generation of IL-10 in vivo. These results suggest that in addition to their beneficial homeostatic influence, antiinflammatory programs activated by efferocytosis in the lung have the undesirable potential to dampen innate antimicrobial responses. They also identify an opportunity to reduce the incidence and severity of pneumonia in the setting of lung injury by pharmacologically targeting synthesis of PGE(2) or ligation of EP2. PMID- 19124659 TI - Postantibiotic effects of telavancin against 16 gram-positive organisms. AB - The in vitro postantibiotic effects (PAEs), postantibiotic sub-MIC effects (PA SMEs), and sub-MIC effects of telavancin were determined for 16 gram-positive organisms. Telavancin staphylococcal, streptococcal, and enterococcal PAE ranges were 0.9 to 3.9 h, 0.4 to 6.7 h, and 0.3 to 2.2 h, respectively. The PA-SME ranges (0.4 times the MIC) for staphylococci, streptococci, and enterococci were 6.7 to >10.7 h, >10.7 to >11.0 h, and >10 to >10.8 h, respectively. The extended PAE of telavancin, together with its long elimination half-life in humans, supports once-daily dosing for this investigational drug. PMID- 19124658 TI - CYP2B6 G516T polymorphism but not rifampin coadministration influences steady state pharmacokinetics of efavirenz in human immunodeficiency virus-infected patients in South India. AB - The dose of efavirenz during concomitant rifampin (RMP) administration is a matter of debate. We studied the influence of RMP coadministration on the steady state pharmacokinetics of efavirenz in human immunodeficiency virus type 1 (HIV 1)-infected patients in South India. Fifty-seven HIV-tuberculosis (TB)-coinfected and 15 HIV-1-infected patients receiving combination antiretroviral therapy (CART) with an efavirenz (600 mg once daily)-containing regimen were recruited. HIV-TB-coinfected patients were receiving treatment with RMP-containing regimens. A complete pharmacokinetic study was conducted with 19 HIV-TB patients on two occasions (with and without RMP). Trough concentrations of efavirenz were measured in the remaining 38 patients during RMP coadministration. The 15 HIV infected patients underwent complete pharmacokinetic sampling on one occasion. Plasma efavirenz was estimated by high-performance liquid chromatography, and genotyping of CYP2B6 G516T polymorphism was performed by sequencing. Peak and trough concentrations and exposure to efavirenz were significantly higher in TT than in GT and GG genotype patients (P < 0.001). Although RMP coadministration decreased the peak and trough concentrations and exposure to efavirenz by 17.8, 20.4, and 18.6%, respectively, the differences were not statistically significant. The trough concentration of efavirenz was subtherapeutic (less than 1.0 microg/ml) in 6 (8%) of 72 patients. In this South Indian population of HIV infected patients, CYP2B6 G516T polymorphism but not RMP coadministration significantly influenced the pharmacokinetics of efavirenz; patients with the TT genotype had very high blood levels of efavirenz. While a small proportion of patients had subtherapeutic efavirenz levels, the clinical implications are uncertain, as all had good immunological responses to CART. PMID- 19124660 TI - Detection of molecular markers of antiviral resistance in influenza A (H5N1) viruses using a pyrosequencing method. AB - Resistance of influenza viruses to antiviral drugs can emerge following medication or may result from natural variation. Two classes of anti-influenza virus drugs targeting either the M2 protein (amantadine and rimantadine) or neuraminidase (NA; oseltamivir and zanamivir) are currently licensed. These drugs are expected to be important in controlling the early stages of a potential pandemic. In the present study, we describe how a pyrosequencing method can be used to rapidly detect established molecular markers of resistance to M2 blockers and NA inhibitors in influenza A (H5N1) viruses. The residues L26, V27, A30, S31, and G34 in the M2 protein were targeted for pyrosequencing. The NA residues for pyrosequencing analysis included the established markers of drug resistance (H274 and N294), as well as residues of less certain relevance (V116, I117, Q136, K150, and I222). A single pair of pyro-reverse transcription (RT)-PCR primers was designed to allow amplification of an approximately 600-nucleotide-long amplicon of the NA genes of H5N1 viruses from various clades/subclades associated with infections in humans. The sensitivity of the assay was demonstrated by the successful pyrosequencing of RNA extracted from samples of serially diluted (10( 5) to 10(-7)) virus stocks with initial concentrations ranging from 10(5) to 10(8) PFU/ml. The markers of resistance were detected in samples with threshold cycle values ranging from 32 to 37, as determined by real-time RT-PCR. The pyrosequencing approach may provide a valuable tool for rapid detection of markers of drug resistance in H5N1 viruses and facilitate the elucidation of the role of such changes in natural and acquired drug resistance. PMID- 19124661 TI - Oral and parenteral therapeutic options for outpatient urinary infections caused by enterobacteriaceae producing CTX-M extended-spectrum beta-lactamases. AB - Effective therapeutic options are needed for community-onset urinary tract infections due to Escherichia coli strains that produce CTX-M extended-spectrum beta-lactamases. We examined 46 urinary isolates producing CTX-M against several oral or long-acting parenteral antimicrobial agents. Approximately 90% were susceptible to fosfomycin and to a combination of cefdinir plus amoxicillin clavulanate. All were susceptible to ertapenem. PMID- 19124662 TI - Contribution of vraSR and graSR point mutations to vancomycin resistance in vancomycin-intermediate Staphylococcus aureus. AB - We describe here the genetic analysis of a vancomycin-susceptible Staphylococcus aureus (VSSA) strain, Mu50Omega, a strain related to vancomycin-intermediate S. aureus (VISA) strain Mu50. Using a combination of Mu50Omega whole-genome sequencing and genome engineering, we observed a stepwise evolution of vancomycin resistance from VSSA to VISA after the mutated vraS and graR genes of Mu50 were engineered into Mu50Omega. PMID- 19124663 TI - Quinolone resistance in Neisseria gonorrhoeae: rapid genotyping of quinolone resistance-determining regions in gyrA and parC genes by melting curve analysis predicts susceptibility. AB - We report a duplex real-time PCR assay for the simultaneous screening of mutations involved in fluoroquinolone resistance within gyrA and parC quninolone resistance-determining regions (QRDRs) in Neisseria gonorrhoeae. Our assay clearly detects all mutated QRDRs and allows the identification of common genotypes, whether the QRDRs contain single or double mutations, providing valuable epidemiological tools. When this method is used in conjunction with similar assays and in vitro analyses, essential antibiotic resistance surveillance can be performed for public health purposes. PMID- 19124664 TI - Activities of dalbavancin against a worldwide collection of 81,673 gram-positive bacterial isolates. AB - Dalbavancin, a long-acting lipoglycopeptide, was evaluated against 81,673 isolates of staphylococci, enterococci, and streptococci collected from 33 countries during worldwide resistance surveillance (2002 to 2007). Regardless of susceptibility to oxacillin, comparable potencies for dalbavancin against Staphylococcus aureus and coagulase-negative staphylococci from all countries were noted (MIC(90), 0.06 to 0.12 microg/ml). Vancomycin-susceptible Enterococcus spp. had dalbavancin MIC(90)s comparable to those for staphylococci, whereas vancomycin-resistant strains were more resistant (MIC(50), >4 microg/ml). beta Hemolytic and viridians group streptococci were very susceptible to dalbavancin (MIC(90), or=16-fold more active than vancomycin against the monitored gram-positive species. PMID- 19124665 TI - Treatment with the fusion inhibitor enfuvirtide influences the appearance of mutations in the human immunodeficiency virus type 1 regulatory protein rev. AB - The gp41-encoding sequence of the env gene contains in two separate regions the Rev-responsive elements (RRE) and the alternative open reading frame of the second exon of the regulatory protein Rev. The binding of Rev to the RRE allows the transport of unspliced/singly spliced viral mRNAs out of the nucleus, an essential step in the life cycle of human immunodeficiency virus type 1 (HIV-1). In this study, we have investigated whether the fusion-inhibitor enfuvirtide (ENF) can induce mutations in Rev and if these mutations correlate with the classical ENF resistance gp41 mutations and with viremia and CD4 cell count. Specific Rev mutations were positively associated with ENF treatment and significantly correlated with classical ENF resistance gp41 mutations. In particular, a cluster was observed for the Rev mutations E57A (E57A(rev)) and N86S(rev) with the ENF resistance gp41 mutations Q40H (Q40H(gp41)) and L45M(gp41). In addition, the presence at week 48 of the E57A(rev) correlates with a significant viremia increase from baseline to week 48 and with a CD4 cell count loss from baseline to week 48. By modeling the RRE structure, we found that the Q40(gp41) and L45(gp41) codons form complementary base pairs in a region of the RRE involved in Rev binding. The conformation of this Rev-binding site is disrupted when Q40H(gp41) and L45M(gp41) occur alone while it is restored when both mutations are present. In conclusion, our study shows that ENF pressure may also affect both Rev and RRE structures and can provide an excellent example of compensatory evolution. This highlights the multiple roles of ENF (and perhaps other entry inhibitors) in modulating the correct interplay between the different HIV-1 genes and proteins during the HIV-1 life cycle. PMID- 19124667 TI - Partnering to reduce risks and improve cardiovascular outcomes: American Heart Association initiatives in action for consumers and patients. PMID- 19124666 TI - A canonical promoter organization of the transcription machinery and its regulators in the Saccharomyces genome. AB - The predominant organizational theme by which the transcription machinery and chromatin regulators are positioned within promoter regions or throughout genes in a genome is largely unknown. We mapped the genomic location of diverse representative components of the gene regulatory machinery in Saccharomyces cerevisiae to an experimental resolution of <40 bp. Sequence-specific gene regulators, chromatin regulators, mediator, and RNA polymerase (Pol) II were found primarily near the downstream border from the "-1" nucleosome, which abuts against the approximately 140-bp nucleosome-free promoter region (NFR). General transcription factors TFIIA, -B, -D, -E, -F, -H were located near the downstream edge from the NFR. The -1 nucleosome dissociated upon Pol II recruitment, but not upon recruitment of only TBP and TFIIB. The position of many sequence-specific regulators in promoter regions correlated with the position of specific remodeling complexes, potentially reflecting functional interactions. Taken together the findings suggest that the combined action of activators and chromatin remodeling complexes remove the -1 nucleosome after the preinitiation complex (PIC) has partially assembled, but before or concomitant with Pol II recruitment. We find PIC assembly, which includes Pol II recruitment, to be a significant rate-limiting step during transcription, but that additional gene specific rate-limiting steps associated with Pol II occur after recruitment. PMID- 19124668 TI - Images in cardiovascular medicine. Continuous murmur after ascending aortic surgery. PMID- 19124669 TI - Letter by Kass regarding article, "Role of left ventricular stiffness in heart failure with normal ejection fraction". PMID- 19124670 TI - Letter by Kessler regarding article, "Effect of short call admissions on length of stay and quality of care for acute decompensated heart failure". PMID- 19124671 TI - Letter by Kleemann and Seidl regarding article, "Necessity for surgical revision of defibrillator leads implanted long-term: causes and management". PMID- 19124673 TI - Neurogenic orthostatic hypotension: a pathophysiological approach. PMID- 19124674 TI - The Swan-Ganz catheters: past, present, and future. A viewpoint. AB - The Swan-Ganz balloon flotation catheter was introduced in 1970. It can be placed at the bedside within a few minutes even in critically ill patients. Although placement of these catheters is not difficult, some training and experience are required to avoid complications and for proper interpretation of the hemodynamic data that can be obtained by pulmonary artery catheterization. Because of the many advantages of balloon flotation catheters compared with conventional catheters, they have been used without a proper indication and frequently overused in critical care units, resulting in many complications, including mortality. The prospective randomized trials have reported that in the majority of clinical circumstances, the routine use of balloon flotation catheters is not indicated. These results are not surprising because balloon flotation catheters are diagnostic and not therapeutic tools. That we have learned a great deal about hemodynamics in critically ill patients with the use of balloon flotation catheters should not be ignored or forgotten. Furthermore, our clinical knowledge of hemodynamics has been made possible because of extensive experience gained from directly determined hemodynamics with the use of balloon flotation catheters. It should also be realized that despite the introduction and refinement of newer noninvasive imaging modalities, a number of clinical circumstances exist in which determination of hemodynamics with the use of a balloon flotation catheter is necessary and should be considered, but only by experienced physicians. With the proper use of Swan-Ganz catheters, our knowledge of hemodynamics has been enhanced considerably. Its abuse, particularly by relatively inexperienced operators, has resulted in serious complications, including death. Prospective randomized clinical trials have demonstrated that the routine use of Swan-Ganz catheters does not provide any benefit. However, use of the Swan-Ganz catheter is still indicated in many situations. PMID- 19124675 TI - Images in cardiovascular medicine. Claudication of the foot in a rugby player. PMID- 19124676 TI - Aliskiren therapy will have minimal effect on intracellular renin of renin producing cells. PMID- 19124677 TI - The chicken and the egg: sympathetic nervous system activity and left ventricular diastolic dysfunction. PMID- 19124679 TI - Sympathetic and baroreflex cardiovascular control in hypertension-related left ventricular dysfunction. AB - The sympathetic overdrive that characterizes essential hypertension is potentiated when left ventricular hypertrophy or congestive heart failure is detected. No information exists, however, on whether this is the case also for left ventricular diastolic dysfunction. In 17 untreated hypertensive subjects with left ventricular diastolic dysfunction (age: 47.7+/-2.9 years, mean+/-SEM), we measured sympathetic nerve traffic (microneurography), heart rate (ECG), and beat-to-beat arterial blood pressure (Finapres) at rest and during baroreceptor deactivation and stimulation. Data were compared with those collected in 20 age matched normotensive and 20 hypertensive subjects without a diastolic function impairment. Muscle sympathetic nerve traffic values were markedly and significantly greater in the 2 hypertensive groups than in the normotensive one (55.3+/-1.2 and 71.2+/-1.6 versus 41.7+/-1.0 bursts per 100 heartbeats, respectively; P<0.01 for both). For a similar blood pressure elevation, however, the sympathetic nerve traffic increase was significantly greater in patients with than without left ventricular diastolic dysfunction (+28.9%; P<0.05). In the population as a whole, muscle sympathetic nerve traffic was significantly and inversely related to various echocardiographic indices of diastolic function. Although baroreflex-heart rate control was significantly attenuated in the 2 hypertensive groups, baroreflex-sympathetic modulation was impaired only in those with diastolic dysfunction. These data provide the first evidence that, in hypertension, activation of the sympathetic nervous system may contribute not only at the blood pressure elevation but also at the development of left ventricular diastolic dysfunction. The sympathetic overactivity, which is likely to be related to the baroreflex impairment, may account for the increased cardiovascular risk characterizing diastolic dysfunction. PMID- 19124678 TI - Angiotensin II type 1 receptor-mediated reduction of angiotensin-converting enzyme 2 activity in the brain impairs baroreflex function in hypertensive mice. AB - Angiotensin-converting enzyme 2 (ACE2), a new component of the brain renin angiotensin system, has been suggested to participate in the central regulation of blood pressure (BP). To clarify the relationship between ACE2 and other brain renin-angiotensin system components, we hypothesized that central angiotensin II type 1 receptors reduce ACE2 expression/activity in hypertensive mice, thereby impairing baroreflex function and promoting hypertension. To test this hypothesis, chronically hypertensive mice (RA) with elevated angiotensin II levels were treated with losartan (angiotensin II type 1 receptor blocker) or PD123319 (angiotensin II type 2 antagonist; 10 mg/kg per day, SC) for 2 weeks. Baseline spontaneous baroreflex sensitivity and brain ACE2 activity were dramatically decreased in RA compared with nontransgenic mice, whereas peripheral ACE2 activity/expression remained unaffected. Losartan, but not PD123319, increased central ACE2 activity, spontaneous baroreflex sensitivity, and normalized BP in RA mice. To confirm the critical role of central ACE2 in BP regulation, we generated a triple-transgenic model with brain ACE2 overexpression on a hypertensive RA background. Triple-transgenic-model mice exhibit lower BP and blunted water intake versus RA, suggesting lower brain angiotensin II levels. Moreover, the impaired spontaneous baroreflex sensitivity, parasympathetic tone, and increased sympathetic drive, observed in RA, were normalized in triple transgenic-model mice. These data suggest that angiotensin II type 1 receptors inhibit ACE2 activity in RA mice brain, thus contributing to the maintenance of hypertension. In addition, overexpression of ACE2 in the brain reduces hypertension by improving arterial baroreflex and autonomic function. Together, our data suggest that angiotensin II type 1 receptor-mediated ACE2 inhibition impairs baroreflex function and support a critical role for ACE2 in the central regulation of BP and the development of hypertension. PMID- 19124680 TI - Diabetes mellitus and 24-hour ambulatory blood pressure monitoring: broadening horizons of risk assessment. PMID- 19124681 TI - Ambulatory blood pressure monitoring and all-cause mortality in elderly people with diabetes mellitus. AB - In a multiethnic cohort of older people with diabetes (n=1178), we assessed whether ambulatory blood pressure (BP) monitoring improves prediction of all cause mortality and cardiovascular mortality when added to baseline covariates, including office BP and heart rate (HR). Secondary analyses assessed whether albuminuria may mediate the association of pulse pressure with mortality. The ambulatory arterial stiffness index was calculated as "1-slope" from the within person regression of diastolic-on-systolic ambulatory BP readings. Mean follow-up was 6.6+/-0.4 years. There were 287 deaths; death certificates were available for 215 deaths (75%), and 110 of them were deemed of cardiovascular cause. Cox models were built incrementally. First, models using clinical and laboratory variables selected albuminuria and office HRs as independent predictors of all-cause and cardiovascular mortality. When ambulatory monitoring data were added, sleep:wake HR ratio and ambulatory arterial stiffness index added significantly to the prediction of all-cause mortality, but only sleep:wake HR ratio added to the prediction of cardiovascular mortality. Office HR and albuminuria retained significance as predictors of both types of mortality. Secondary analyses without adjustment for albuminuria confirmed the predictive value of office HR and sleep/wake HR, whereas 24-hour pulse pressure and sleep systolic BP were also independently predictive of all-cause and cardiovascular mortality, respectively. In conclusion, office HR and albuminuria were strong predictors of mortality. Ambulatory monitoring improved the prediction of risk through its assessment of sleep HR dipping and of ambulatory arterial stiffness index, a measure of the dynamic relationship between systolic and diastolic BPs. Albuminuria may mediate the association between BP and mortality. PMID- 19124682 TI - Plasma kallikrein mediates angiotensin II type 1 receptor-stimulated retinal vascular permeability. AB - Hypertension is a leading risk factor for the development and progression of diabetic retinopathy and contributes to a variety of other retinal diseases in the absence of diabetes mellitus. Inhibition of the renin-angiotensin system has been shown to provide beneficial effects against diabetic retinopathy, both in the absence and presence of hypertension, suggesting that angiotensin II (Ang II) and the Ang II type 1 receptor may contribute to retinal vascular dysfunction. We investigated the effects of the Ang II type 1 receptor antagonist candesartan on retinal vascular permeability (RVP) in normotensive rats with streptozotocin induced diabetes mellitus and in rats with Ang II-induced hypertension. We showed that candesartan treatment decreased diabetes mellitus- and Ang II-stimulated RVP by 58% (P<0.05) and 79% (P<0.05), respectively, compared with untreated controls, suggesting that activation of the Ang II type 1 receptor contributes to blood retinal barrier dysfunction. We found that plasma kallikrein levels are increased in the retina of rats with Ang II-stimulated hypertension and that intravitreal injection of either plasma kallikrein or bradykinin is sufficient to increase RVP. We showed that a novel small molecule inhibitor of plasma kallikrein, 1 benzyl-1H-pyrazole-4-carboxylic acid 4-carbamimidoyl-benzylamide, delivered systemically via a subcutaneous pump, decreased Ang II-stimulated RVP by 70% (P<0.05) and ameliorates Ang II-induced hypertension, measured from the carotid artery by telemetry, but did not reduce Ang II-induced retinal leukostasis. These findings demonstrate that activation of the Ang II type 1 receptor increases RVP and suggest that systemic plasma kallikrein inhibition may provide a new therapeutic approach for ameliorating blood-retinal barrier dysfunction induced by hypertension. PMID- 19124683 TI - The convalescent. PMID- 19124684 TI - Elevated striatal dopamine function linked to prodromal signs of schizophrenia. AB - CONTEXT: A major limitation on the development of biomarkers and novel interventions for schizophrenia is that its pathogenesis is unknown. Although elevated striatal dopamine activity is thought to be fundamental to schizophrenia, it is unclear when this neurochemical abnormality develops in relation to the onset of illness and how this relates to the symptoms and neurocognitive impairment seen in individuals with prodromal symptoms of schizophrenia. OBJECTIVES: To determine whether striatal dopamine function is elevated in individuals with prodromal symptoms of schizophrenia before the onset of psychosis and to assess how this relates to the symptoms and neurocognitive impairment. DESIGN: Case-control study of in vivo striatal dopaminergic function. SETTING: Academic research. Patients Patients were recruited from a community mental health service. Twenty-four patients having prodromal symptoms of schizophrenia were compared with 7 patients having schizophrenia and with 12 matched healthy control subjects from the same community. Main Outcome Measure Striatal 6-fluoro-l-dopa F 18-dopa uptake measured using positron emission tomographic (18)F-dopa imaging. RESULTS: Striatal (18)F-dopa uptake was elevated in patients with prodromal symptoms of schizophrenia (effect size, 0.75) to an intermediate degree compared with that in patients with schizophrenia (effect size, 1.25). The elevation was localized in the associative striatum in both groups. Moreover, striatal (18)F-dopa uptake in patients with prodromal symptoms of schizophrenia was correlated with the severity of prodromal psychopathologic and neuropsychological impairment but not with the severity of anxiety or depressive symptoms. CONCLUSIONS: These findings indicate that dopamine overactivity predates the onset of schizophrenia in individuals with prodromal psychotic symptoms, is predominantly localized in the associative striatum, and is correlated with the severity of symptoms and neurocognitive dysfunction. PMID- 19124685 TI - Alternative splicing, methylation state, and expression profile of tropomyosin related kinase B in the frontal cortex of suicide completers. AB - CONTEXT: Although most of the effort to understand the neurobiology of depressive states and suicide has focused on neuronal processes, recent studies suggest that astroglial dysfunction may play an important role. A truncated variant of the tropomyosin-related kinase B (TrkB.T1) is expressed in astrocytes, and brain derived neurotrophic factor-TrkB signaling has been linked to mood disorders. OBJECTIVE: To test the hypothesis that TrkB.T1 expression is downregulated in suicide completers and that this downregulation is mediated by an epigenetic process. DESIGN: Postmortem case-control study. Patients, Setting, and MAIN OUTCOME MEASURES: Thirty-nine French Canadian men underwent screening at the Douglas Hospital Research Institute using the HG-U133 plus 2 microarray chip. Nine frontal cortical regions and the cerebellum were assessed using a microarray screening approach for extreme expression differences across subjects and a conventional screening approach. Results were validated by quantitative polymerase chain reaction and Western blot analyses. Animal experiments were performed to control for drug and alcohol effects. Genetic and epigenetic studies were performed by means of direct sequencing and bisulfite mapping. RESULTS: We found that 10 of 28 suicide completers (36%) demonstrated significant decreases in different probe sets specific to TrkB.T1 in Brodmann areas 8 and 9. These findings were generalizable to other frontal regions but not to the cerebellum. The decrease in TrkB expression was specific to the T1 splice variant. Our results were not accounted for by substance comorbidity or by reduction in astrocyte number. We found no effect of genetic variation in a 2500-base pair promoter region or at relevant splice junctions; however, we detected an effect of methylation state at particular CpG dinucleotides on TrkB.T1 expression. CONCLUSION: A reduction of TrkB.T1 expression in the frontal cortex of a subpopulation of suicide completers is associated with the methylation state of the promoter region. PMID- 19124686 TI - Effects of HTR1A C(-1019)G on amygdala reactivity and trait anxiety. AB - CONTEXT: Serotonin 1A (5-hydroxytryptamine 1A [5-HT(1A)]) autoreceptors mediate negative feedback inhibition of serotonergic neurons and play a critical role in regulating serotonin signaling involved in shaping the functional response of major forebrain targets, such as the amygdala, supporting complex behavioral processes. A common functional variation (C[-1019]G) in the human 5-HT(1A) gene (HTR1A) represents 1 potential source of such interindividual variability. Both in vitro and in vivo, -1019G blocks transcriptional repression, leading to increased autoreceptor expression. Thus, -1019G may contribute to relatively decreased serotonin signaling at postsynaptic forebrain target sites via increased negative feedback. OBJECTIVES: To evaluate the effects of HTR1A C( 1019)G on amygdala reactivity and to use path analyses to explore the impact of HTR1A-mediated variability in amygdala reactivity on individual differences in trait anxiety. We hypothesized that -1019G, which potentially results in decreased serotonin signaling, would be associated with relatively decreased amygdala reactivity and related trait anxiety. DESIGN: Imaging genetics in participants from an archival database. PARTICIPANTS: Eighty-nine healthy adults. RESULTS: Consistent with prior findings, -1019G was associated with significantly decreased threat-related amygdala reactivity. Importantly, this effect was independent of that associated with another common functional polymorphism that affects serotonin signaling, 5-HTTLPR. While there were no direct genotype effects on trait anxiety, HTR1A C(-1019)G indirectly predicted 9.2% of interindividual variability in trait anxiety through its effects on amygdala reactivity. CONCLUSIONS: Our findings further implicate relatively increased serotonin signaling, associated with a genetic variation that mediates increased 5-HT(1A) autoreceptors, in driving amygdala reactivity and trait anxiety. Moreover, they provide empirical documentation of the basic premise that genetic variation indirectly affects emergent behavioral processes related to psychiatric disease risk by biasing the response of underlying neural circuitries. PMID- 19124687 TI - Influence of functional variant of neuronal nitric oxide synthase on impulsive behaviors in humans. AB - CONTEXT: Human personality is characterized by substantial heritability but few functional gene variants have been identified. Although rodent data suggest that the neuronal isoform of nitric oxide synthase (NOS-I) modifies diverse behaviors including aggression, this has not been translated to human studies. OBJECTIVES: To investigate the functionality of an NOS1 promoter repeat length variation (NOS1 Ex1f variable number tandem repeat [VNTR]) and to test whether it is associated with phenotypes relevant to impulsivity. DESIGN: Molecular biological studies assessed the cellular consequences of NOS1 Ex1f VNTR; association studies were conducted to investigate the impact of this genetic variant on impulsivity; imaging genetics was applied to determine whether the polymorphism is functional on a neurobiological level. SETTING: Three psychiatric university clinics in Germany. PARTICIPANTS: More than 3200 subjects were included in the association study: 1954 controls, 403 patients with personality disorder, 383 patients with adult attention-deficit/hyperactivity disorder (ADHD), 151 with familial ADHD, 189 suicide attempters, and 182 criminal offenders. MAIN OUTCOME MEASURES: For the association studies, the major outcome criteria were phenotypes relevant to impulsivity, namely, the dimensional phenotype conscientiousness and the categorical phenotypes adult ADHD, aggression, and cluster B personality disorder. RESULTS: A novel functional promoter polymorphism in NOS1 was associated with traits related to impulsivity, including hyperactive and aggressive behaviors. Specifically, the short repeat variant was more frequent in adult ADHD, cluster B personality disorder, and autoaggressive and heteroaggressive behavior. This short variant came along with decreased transcriptional activity of the NOS1 exon 1f promoter and alterations in the neuronal transcriptome including RGS4 and GRIN1. On a systems level, it was associated with hypoactivation of the anterior cingulate cortex, which is involved in the processing of emotion and reward in behavioral control. CONCLUSION: These findings implicate deficits in neuronal signaling via nitric oxide in moderation of prefrontal circuits underlying impulsivity-related behavior in humans. PMID- 19124689 TI - A genetically informed study of the association between childhood separation anxiety, sensitivity to CO(2), panic disorder, and the effect of childhood parental loss. AB - CONTEXT: Childhood separation anxiety disorder can predate panic disorder, which usually begins in early adulthood. Both disorders are associated with heightened sensitivity to inhaled CO(2) and can be influenced by childhood parental loss. OBJECTIVES: To find the sources of covariation between childhood separation anxiety disorder, hypersensitivity to CO(2), and panic disorder in adulthood and to measure the effect of childhood parental loss on such covariation. DESIGN: Multivariate twin study. PARTICIPANTS: Seven hundred twelve young adults from the Norwegian Institute of Public Health Twin Panel, a general population cohort. MAIN OUTCOME MEASURES: Personal direct assessment of lifetime panic disorder through structured psychiatric interviews, history of childhood parental loss, and separation anxiety disorder symptoms. Subjective anxiety response to a 35% CO(2)/65% O(2) inhaled mixture compared with compressed air (placebo). RESULTS: Our best-fitting solution yielded a common pathway model, implying that covariation between separation anxiety in childhood, hypersensitivity to CO(2), and panic disorder in adulthood can be explained by a single latent intervening variable influencing all phenotypes. The latent variable governing the 3 phenotypes' covariation was in turn largely (89%) influenced by genetic factors and childhood parental loss (treated as an identified element of risk acting at a family-wide level), which accounted for the remaining 11% of covariance. Residual variance was explained by 1 specific genetic variance component for separation anxiety disorder and variable-specific unique environmental variance components. CONCLUSIONS: Shared genetic determinants appear to be the major underlying cause of the developmental continuity of childhood separation anxiety disorder into adult panic disorder and the association of both disorders with heightened sensitivity to CO(2). Inasmuch as childhood parental loss is a truly environmental risk factor, it can account for a significant additional proportion of the covariation of these 3 developmentally related phenotypes. PMID- 19124688 TI - Deficient activity in the neural systems that mediate self-regulatory control in bulimia nervosa. AB - CONTEXT: Disturbances in neural systems that mediate voluntary self-regulatory processes may contribute to bulimia nervosa (BN) by releasing feeding behaviors from regulatory control. OBJECTIVE: To study the functional activity in neural circuits that subserve self-regulatory control in women with BN. DESIGN: We compared functional magnetic resonance imaging blood oxygenation level-dependent responses in patients with BN with healthy controls during performance of the Simon Spatial Incompatibility task. SETTING: University research institute. PARTICIPANTS: Forty women: 20 patients with BN and 20 healthy control participants. Main Outcome Measure We used general linear modeling of Simon Spatial Incompatibility task-related activations to compare groups on their patterns of brain activation associated with the successful or unsuccessful engagement of self-regulatory control. RESULTS: Patients with BN responded more impulsively and made more errors on the task than did healthy controls; patients with the most severe symptoms made the most errors. During correct responding on incongruent trials, patients failed to activate frontostriatal circuits to the same degree as healthy controls in the left inferolateral prefrontal cortex (Brodmann area [BA] 45), bilateral inferior frontal gyrus (BA 44), lenticular and caudate nuclei, and anterior cingulate cortex (BA 24/32). Patients activated the dorsal anterior cingulate cortex (BA 32) more when making errors than when responding correctly. In contrast, healthy participants activated the anterior cingulate cortex more during correct than incorrect responses, and they activated the striatum more when responding incorrectly, likely reflecting an automatic response tendency that, in the absence of concomitant anterior cingulate cortex activity, produced incorrect responses. CONCLUSIONS: Self-regulatory processes are impaired in women with BN, likely because of their failure to engage frontostriatal circuits appropriately. These findings enhance our understanding of the pathogenesis of BN by pointing to functional abnormalities within a neural system that subserves self-regulatory control, which may contribute to binge eating and other impulsive behaviors in women with BN. PMID- 19124690 TI - Childhood trauma and risk for chronic fatigue syndrome: association with neuroendocrine dysfunction. AB - CONTEXT: Childhood trauma appears to be a potent risk factor for chronic fatigue syndrome (CFS). Evidence from developmental neuroscience suggests that early experience programs the development of regulatory systems that are implicated in the pathophysiology of CFS, including the hypothalamic-pituitary-adrenal axis. However, the contribution of childhood trauma to neuroendocrine dysfunction in CFS remains obscure. OBJECTIVES: To replicate findings on the relationship between childhood trauma and risk for CFS and to evaluate the association between childhood trauma and neuroendocrine dysfunction in CFS. DESIGN, SETTING, AND PARTICIPANTS: A case-control study of 113 persons with CFS and 124 well control subjects identified from a general population sample of 19 381 adult residents of Georgia. MAIN OUTCOME MEASURES: Self-reported childhood trauma (sexual, physical, and emotional abuse; emotional and physical neglect), psychopathology (depression, anxiety, and posttraumatic stress disorder), and salivary cortisol response to awakening. RESULTS: Individuals with CFS reported significantly higher levels of childhood trauma and psychopathological symptoms than control subjects. Exposure to childhood trauma was associated with a 6-fold increased risk of CFS. Sexual abuse, emotional abuse, and emotional neglect were most effective in discriminating CFS cases from controls. There was a graded relationship between exposure level and CFS risk. The risk of CFS conveyed by childhood trauma further increased with the presence of posttraumatic stress disorder symptoms. Only individuals with CFS and with childhood trauma exposure, but not individuals with CFS without exposure, exhibited decreased salivary cortisol concentrations after awakening compared with control subjects. CONCLUSIONS: Our results confirm childhood trauma as an important risk factor of CFS. In addition, neuroendocrine dysfunction, a hallmark feature of CFS, appears to be associated with childhood trauma. This possibly reflects a biological correlate of vulnerability due to early developmental insults. Our findings are critical to inform pathophysiological research and to devise targets for the prevention of CFS. PMID- 19124691 TI - Influence of cognitive status, age, and APOE-4 genetic risk on brain FDDNP positron-emission tomography imaging in persons without dementia. AB - CONTEXT: Amyloid senile plaques and tau neurofibrillary tangles are neuropathological hallmarks of Alzheimer disease that accumulate in the brains of people without dementia years before they develop dementia. Positron emission tomography (PET) scans after intravenous injections of 2-(1-{6-[(2-[F 18]fluoroethyl)(methyl)amino]-2-naphthyl}ethylidene)malononitrile (FDDNP), which binds to plaques and tangles in vitro, demonstrate increased cerebral binding in patients with Alzheimer disease compared with cognitively intact controls. Here we investigated whether known risk factors for Alzheimer disease and dementia are associated with FDDNP-PET binding. OBJECTIVE: To determine if impaired cognitive status, older age, apolipoprotein E-4 (APOE-4) genetic risk for Alzheimer disease, family history of dementia, and less education are associated with increased regional cerebral FDDNP-PET binding. DESIGN: Cross-sectional clinical study. SETTING: A university research institute. PARTICIPANTS: Volunteer sample of 76 middle-aged and older persons without dementia (mean age, 67 years) including 36 with mild cognitive impairment. Of the 72 subjects with genetic data, 34 were APOE-4 carriers. MAIN OUTCOME MEASURES: The FDDNP-PET signal in brain regions of interest, including medial and lateral temporal, posterior cingulate, parietal, and frontal. RESULTS: For all regions studied, cognitive status was associated with increased FDDNP binding (P < .02 to .005). Older age was associated with increased lateral temporal FDDNP binding. Carriers of APOE-4 demonstrated higher frontal FDDNP binding than noncarriers. In the mild cognitive impairment group, age was associated with increased medial and lateral temporal FDDNP binding, and APOE-4 carriers had higher medial temporal binding than noncarriers. CONCLUSIONS: Impaired cognitive status, older age, and APOE-4 carrier status are associated with increased brain FDDNP-PET binding in persons without dementia, consistent with previous clinical and postmortem studies associating these risk factors with amyloid plaque and tau tangle accumulation. Stratifying subject groups according to APOE-4 carrier status, age, and cognitive status may therefore be an informative strategy in future clinical trials using FDDNP-PET. PMID- 19124692 TI - Real-time electronic diary reports of cue exposure and mood in the hours before cocaine and heroin craving and use. AB - CONTEXT: In ecological momentary assessment (EMA), participants electronically report their activities and moods in their daily environments in real time, enabling a truly prospective approach to the study of acute precipitants of behavioral events. Ecological momentary assessment has greatly enhanced the study of tobacco addiction, but its use has rarely been attempted in individuals with cocaine or heroin addiction. OBJECTIVE: To prospectively monitor the acute daily life precipitants of craving for and use of cocaine and heroin. DESIGN: Cohort study. PARTICIPANTS: A volunteer sample of 114 cocaine- and heroin-abusing outpatients who were being treated with methadone provided EMA data on handheld electronic devices for 14 918 person-days (mean, 130.9; range, 6-189 days per participant). Of these outpatients, a total of 102 (63 men, 39 women) provided acute precraving and/or preuse data and were thus included in the present analyses. MAIN OUTCOME MEASURES: Changes in reports of mood and exposure to 12 putative drug-use triggers at random intervals during the 5 hours preceding each self-reported episode of drug craving or use, analyzed via repeated-measures logistic regression (generalized linear mixed models). RESULTS: During the 5 hours preceding cocaine use or heroin craving, most of the 12 putative triggers showed linear increases. Cocaine use was most robustly associated with increases in participants reporting that they "saw [the] drug" (P < .001), were "tempted to use out of the blue" (P < .001), "wanted to see what would happen if I used" (P < .001), and were in a good mood (P < .001). Heroin craving was most robustly associated with increases in reports of feeling sad (P < .001) or angry (P = .01). Cocaine craving and heroin use showed few reliable associations with any of the putative triggers assessed. CONCLUSIONS: These findings confirm that polydrug abusing individuals can provide behavioral data in their daily environments using handheld electronic devices and that those data can reveal orderly patterns, including prospectively detectable harbingers of craving and use, which may differ across drugs. PMID- 19124693 TI - Distinct effects of {delta}9-tetrahydrocannabinol and cannabidiol on neural activation during emotional processing. AB - CONTEXT: Cannabis use can both increase and reduce anxiety in humans. The neurophysiological substrates of these effects are unknown. OBJECTIVE: To investigate the effects of 2 main psychoactive constituents of Cannabis sativa (Delta9-tetrahydrocannabinol [Delta9-THC] and cannabidiol [CBD]) on regional brain function during emotional processing. DESIGN: Subjects were studied on 3 separate occasions using an event-related functional magnetic resonance imaging paradigm while viewing faces that implicitly elicited different levels of anxiety. Each scanning session was preceded by the ingestion of either 10 mg of Delta9-THC, 600 mg of CBD, or a placebo in a double-blind, randomized, placebo controlled design. PARTICIPANTS: Fifteen healthy, English-native, right-handed men who had used cannabis 15 times or less in their life. MAIN OUTCOME MEASURES: Regional brain activation (blood oxygenation level-dependent response), electrodermal activity (skin conductance response [SCR]), and objective and subjective ratings of anxiety. RESULTS: Delta9-Tetrahydrocannabinol increased anxiety, as well as levels of intoxication, sedation, and psychotic symptoms, whereas there was a trend for a reduction in anxiety following administration of CBD. The number of SCR fluctuations during the processing of intensely fearful faces increased following administration of Delta9-THC but decreased following administration of CBD. Cannabidiol attenuated the blood oxygenation level dependent signal in the amygdala and the anterior and posterior cingulate cortex while subjects were processing intensely fearful faces, and its suppression of the amygdalar and anterior cingulate responses was correlated with the concurrent reduction in SCR fluctuations. Delta9-Tetrahydrocannabinol mainly modulated activation in frontal and parietal areas. CONCLUSIONS: Delta9 Tetrahydrocannabinol and CBD had clearly distinct effects on the neural, electrodermal, and symptomatic response to fearful faces. The effects of CBD on activation in limbic and paralimbic regions may contribute to its ability to reduce autonomic arousal and subjective anxiety, whereas the anxiogenic effects of Delta9-THC may be related to effects in other brain regions. PMID- 19124694 TI - Concomitant use of Lorazepam with tamoxifen in bipolar mania clinical trials. PMID- 19124695 TI - Depression in the prodromal phase of Alzheimer disease and the reverse causal hypothesis. PMID- 19124696 TI - Diagnosis of neonatal sepsis using universal primer polymerase chain reaction before and after starting antibiotic drug therapy. AB - OBJECTIVE: To study universal primer 16S rRNA gene polymerase chain reaction (PCR) for diagnosis of blood culture-positive neonatal sepsis before and after starting antibiotic drug therapy. DESIGN: Prospective study of diagnostic tests. SETTING: Level III neonatal intensive care unit. Patients Neonates with a fresh episode of clinically suspected sepsis were enrolled; those with major malformations, life expectancy less than 12 hours, or contaminated blood cultures were excluded. INTERVENTIONS: Before starting antibiotic drug therapy, PCR (0 hour), blood culture, and sepsis screening (complete blood cell counts, micro erythrocyte sedimentation rate, and C-reactive protein level) were performed. The PCR was repeated 12, 24, and 48 hours after starting antibiotic drug therapy. MAIN OUTCOME MEASURES: The primary outcomes were the sensitivity and specificity of 0-hour PCR for diagnosing blood culture-positive sepsis, and the secondary outcome was the proportion of 0-hour PCR-positive patients who remained positive after antibiotic drug therapy. RESULTS: Of 306 patients evaluated, 242 were included (mean [SD] gestation, 32.2 [3.1] weeks; and mean [SD] birth weight, 1529.2 [597.2] g). Blood culture was positive in 52 patients and 0-hour PCR in 57. The sensitivity, specificity, positive and negative predictive values, and positive and negative likelihood ratios of PCR were 96.2%, 96.3%, 87.7%, 98.8%, 26.1, and 0.04, respectively. Two patients were blood culture positive but 0-hour PCR negative, whereas 7 were 0-hour PCR positive but blood culture negative. Of the 0-hour PCR-positive patients, 7 remained positive at 12 hours and none at 24 and 48 hours after starting antibiotic drug therapy. In 0-hour PCR-positive patients, no predictors of positive 12-hour PCR were identified. CONCLUSION: Universal primer PCR can accurately diagnose neonatal sepsis before but not after antibiotic drugs are given. PMID- 19124697 TI - Neonatal sepsis: looking beyond the blood culture: evaluation of a study of universal primer polymerase chain reaction for identification of neonatal sepsis. PMID- 19124698 TI - Cigarette smoking and perception of a movie character in a film trailer. AB - OBJECTIVE: To study the effects of smoking in a film trailer. DESIGN: Experimental study. SETTING: Ten secondary schools in Northern Germany. PARTICIPANTS: A sample of 1051 adolescents with a mean (SD) age of 14.2 (1.8) years. Main Exposures Participants were randomized to view a 42-second film trailer in which the attractive female character either smoked for about 3 seconds or did not smoke. MAIN OUTCOME MEASURES: Perception of the character was measured via an 8-item semantic differential scale. Each item consisted of a polar-opposite pair (eg, "sexy/unsexy") divided on a 7-point scale. Responses to individual items were summed and averaged. This scale was named "attractiveness." The Cronbach alpha for the attractiveness rating was 0.85. RESULTS: Multilevel mixed-effects linear regression was used to test the effect of smoking in a film trailer. Smoking in the film trailer did not reach significance in the linear regression model (z = 0.73; P = .47). Smoking status of the recipient (z = 3.81; P < .001) and the interaction between smoking in the film trailer and smoking status of the recipient (z = 2.21; P = .03) both reached statistical significance. Ever smokers and never smokers did not differ in their perception of the female character in the nonsmoking film trailer. In the smoking film trailer, ever smokers judged the character significantly more attractive than never smokers. CONCLUSION: Even incidental smoking in a very short film trailer might strengthen the attractiveness of smokers in youth who have already tried their first cigarettes. PMID- 19124699 TI - Health care use and health and functional impact of developmental disabilities among US children, 1997-2005. AB - OBJECTIVE: To present nationally representative estimates of health-related limitations, needs, and service use among US children with and without developmental disabilities (DDs). DESIGN: Retrospective analysis of data from a sample of US households from the 1997-2005 National Health Interview Surveys. PARTICIPANTS: Children aged 3 to 17 years (n = 95 132). MAIN OUTCOME MEASURES: Parents or other knowledgeable adults reported on their children's DDs, health needs, and use of health and education services. Developmental disabilities included attention-deficit/hyperactivity disorder, autism, blindness, cerebral palsy, deaf/a lot of trouble hearing, learning disability, mental retardation, seizures, stuttering/stammering, and other developmental delay. RESULTS: Among children with 1 or more DDs, prevalence estimates for limitations in movement (6.1%), needed help with personal care (3.2%), needed special equipment (3.5%), received home health care (1.4%), and regularly took prescription medication(s) (37.5%) were 4 to 32 times higher than for children without DDs. Children with DDs were 2 to 8 times as likely to have had more than 9 health care visits (14.9%), received special education (38.8%), had a surgical or medical procedure (7.5%), and recently visited a medical specialist (23.9%), mental health professional (26.6%), therapist/allied health professional (19.6%), and/or emergency department (10.3%). Effects were generally stable during the study interval and independent of age, race, sex, and family income. Cerebral palsy, autism, mental retardation, blindness, and deafness/a lot of trouble hearing were associated with the highest levels of health and functional impact indicators. CONCLUSIONS: Developmental disabilities profoundly affect children's health and functioning. These data can inform evidence-based targeted prevention strategies for minimizing functional limitations and lifetime disability. Additional study of unmet needs and access to care is needed. PMID- 19124700 TI - Display of health risk behaviors on MySpace by adolescents: prevalence and associations. AB - OBJECTIVE: To determine the prevalence of and associations among displayed risk behavior information that suggests sexual behavior, substance use, and violence in a random sample of the self-reported 18-year-old adolescents' publicly accessible MySpace Web profiles. DESIGN: Cross-sectional study using content analysis of Web profiles between July 15 and September 30, 2007. SETTING: www.MySpace.com. PARTICIPANTS: A total of 500 publicly available Web profiles of self-reported 18-year-olds in the United States. MAIN OUTCOME MEASURES: Prevalence and associations among displayed health risk behaviors, including sexual behavior, substance use, or violence, on Web profiles. RESULTS: A total of 270 (54.0%) profiles contained risk behavior information: 120 (24.0%) referenced sexual behaviors, 205 (41.0%) referenced substance use, and 72 (14.4)% referenced violence. Female adolescents were less likely to display violence references (odds ratio [OR], 0.3; 95% confidence interval [CI], 0.15-0.6). Reporting a sexual orientation other than "straight" was associated with increased display of references to sexual behavior (OR, 4.48; 95% CI, 1.27-15.98). Displaying church or religious involvement was associated with decreased display of all outcomes (sex: OR, 0.32; 95% CI, 0.12-0.86; substance use: OR, 0.38; 95% CI, 0.19-0.79; violence: OR, 0.12; 95% CI, 0.02-0.87; any risk factor: OR, 0.36; 95% CI, 0.19 0.7). Displaying sport or hobby involvement was associated with decreased references to violence (OR, 0.27; 95% CI, 0.09-0.79) and any risk factor (OR, 0.46; 95% CI, 0.27-0.79). CONCLUSIONS: Adolescents frequently display risk behavior information on public Web sites. Further study is warranted to explore the validity of such information and the potential for using social networking Web sites for health promotion. PMID- 19124701 TI - Reducing at-risk adolescents' display of risk behavior on a social networking web site: a randomized controlled pilot intervention trial. AB - OBJECTIVE: To determine whether an online intervention reduces references to sex and substance abuse on social networking Web sites among at-risk adolescents. DESIGN: Randomized controlled intervention trial. SETTING: www.MySpace.com. PARTICIPANTS: Self-described 18- to 20-year-olds with public MySpace profiles who met our criteria for being at risk (N = 190). Intervention Single physician e mail. MAIN OUTCOME MEASURES: Web profiles were evaluated for references to sex and substance use and for security settings before and 3 months after the intervention. RESULTS: Of 190 subjects, 58.4% were male. At baseline, 54.2% of subjects referenced sex and 85.3% referenced substance use on their social networking site profiles. The proportion of profiles in which references decreased to 0 was 13.7% in the intervention group vs 5.3% in the control group for sex (P = .05) and 26.0% vs 22% for substance use (P = .61). The proportion of profiles set to "private" at follow-up was 10.5% in the intervention group and 7.4% in the control group (P = .45). The proportion of profiles in which any of these 3 protective changes were made was 42.1% in the intervention group and 29.5% in the control group (P = .07). CONCLUSIONS: A brief e-mail intervention using social networking sites shows promise in reducing sexual references in the online profiles of at-risk adolescents. Further study should assess how adolescents view different risk behavior disclosures to promote safe use of the Internet. PMID- 19124702 TI - Outbreak of escherichia coli O157: H7 infections after Petting Zoo visits, North Carolina State Fair, October-November 2004. AB - OBJECTIVES: To identify cases, describe the outbreak, implement control measures, and identify factors associated with infection or protection from infection, including contact with animals and hand hygiene practices. DESIGN: Case finding, a case-control study of 45 cases and 188 controls, environmental investigation, and molecular subtyping of clinical and environmental Escherichia coli O157:H7 isolates. SETTING: The 2004 North Carolina State Fair. PARTICIPANTS: Case patients were fair visitors who had laboratory-confirmed E coli O157 infections, hemolytic uremic syndrome (HUS) diagnoses, or bloody diarrheal illnesses. Control subjects were recruited from a randomized list of persons who had purchased fair tickets online. Environmental samples from the fairgrounds were obtained from locations that had held animals during the fair. Main Exposure Visiting a petting zoo. MAIN OUTCOME MEASURES: Case finding: Summary descriptive statistics of suspected, probable, or confirmed E coli O157:H7 infections, signs, symptoms, and HUS. Environmental investigation: E coli O157:H7 isolates, pulsed-field gel electrophoresis patterns, and spatial distribution of source locations. Case control study: Odds ratios (ORs) comparing reported fair-related activities, hygiene practices, and zoonotic disease knowledge with outcome. RESULTS: A total of 108 case patients were ascertained, including 41 with laboratory-confirmed illness and 15 who experienced HUS. Forty-five case patients and 188 controls were enrolled in the case-control study. Visits to a petting zoo having substantial environmental E coli O157:H7 contamination were associated with illness (age-adjusted OR, 8.2; 95% confidence interval [CI], 3.3-20.3). Among children 5 years or younger who had visited the implicated petting zoo, contact with animal manure (OR, 6.9; 95% CI, 2.2-21.9) and hand-to-mouth behaviors (OR, 10.6; 95% CI, 2.0-55.0) were associated with illness. Reported hand hygiene practices did not differ significantly (OR, 1.8; 95% CI, 0.3-9.5). Reported awareness of the risk for zoonotic disease was protective (OR, 0.1; 95% CI, 0.03 0.5). Environmental samples from the petting zoo implicated in the case-control study yielded E coli O157:H7, with indistinguishable pulsed-field gel electrophoresis patterns from the predominant strain. CONCLUSIONS: We describe one of the largest petting zoo outbreaks of E coli O157:H7 to date. Persons became infected after contact with manure and engaging in hand-to-mouth behaviors in a petting zoo having substantial E coli O157:H7 contamination. Use of alcohol based hand-sanitizing gels was not protective, although knowledge of the risk for zoonotic infection was protective. Future investigations in similar outbreaks should assess risks for infection and protective measures (eg, physical barriers separating visitors from animal manure, education, and appropriate hand hygiene practices). PMID- 19124703 TI - Respite care for children with special health care needs. AB - OBJECTIVES: To describe (1) the subpopulation of children with special health care needs (CSHCN) in need of respite care and (2) the factors associated with unmet respite care needs. DESIGN: Cross-sectional study using the 2001 National Survey of Children With Special Health Care Needs. SETTING: The United States. PARTICIPANTS: Caregivers of CSHCN. MAIN OUTCOME MEASURES: We determined the association of sociodemographic factors and health status of CSHCN with need for respite care and unmet needs for respite care. RESULTS: Of the 38 831 respondents, 3178 (8.8%) reported need for respite care in the prior 12 months. The CSHCN who were young and uninsured/had insurance gaps or had public insurance and those with functional limitations and unstable health conditions were more likely to need respite care. Twenty-four percent of CSHCN who needed respite care did not receive such services. In multivariate analysis, higher maternal education (adjusted odds ratio [AOR], 1.6 for more than high school education compared with high school education or less), some and severe functional limitations (AORs, 2.5 and 5.6, respectively, compared with no limitation), and unstable health condition (AOR, 2.3 compared with stable condition) were associated with greater unmet respite care needs. The CSHCN with public insurance were half as likely and CSHCN with private insurance were as likely to report unmet respite care needs as those who were uninsured/with insurance gaps. CONCLUSIONS: Respite care needs of CSHCN are frequently unmet. Furthermore, strategies are needed to improve access to respite care for families of CSHCN. PMID- 19124704 TI - Subjective social status and shaming experiences in relation to adolescent depression. AB - OBJECTIVE: To investigate the associations among social status, shaming experiences, and adolescent depression using a status-shaming model. DESIGN: Population-based, self-report cohort study. SETTING: Vastmanland, Sweden. PARTICIPANTS: A cohort of 5396 students in grade 9 (age 15-16 years) and the second year of high school (age 17-18 years). Intervention Participants completed the anonymous questionnaire Survey of Adolescent Life in Vastmanland-2006 during class hours. MAIN OUTCOME MEASURES: We investigated the prevalence of depression according to the Depression Self-Rating Scale of the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition, Text Revision), A-criterion, regarding subjective social status, shaming experiences, and social background. Social status was measured as (1) attributed status of a family's socioeconomic and social standing and (2) acquired status of peer group and school. Binary logistic regressions were used for the analyses. RESULTS: Shaming experiences and low social status interacted with depression. If shaming experiences were present, participants with both high and low attributed status were at increased risk for depression (odds ratio [low and high groups, respectively], 5.4-6.9), whereas medium status seemed to have a protective function. For acquired status, the highest elevated risk was found in participants with low status (odds ratio [girls and boys, respectively], 6.7-8.6). CONCLUSIONS: Social status may influence the risk for depression when an individual is subjected to shaming experiences. The present study contributes to the mapping of the influence of social status on health and may have essential implications for understanding, preventing, and treating adolescent depression. PMID- 19124705 TI - The validity of teens' and young adults' self-reported condom use. AB - OBJECTIVE: To examine the concordance between teens' and young adults' self reported condom use, assessed by audio-computer-assisted self-interviewing, and Y chromosome polymerase chain reaction (Yc-PCR) assay, a nondisease marker for detecting the presence of sperm in vaginal fluid for 14 days after unprotected vaginal sex. DESIGN: Randomized trial of a human immunodeficiency virus prevention program. Only data from baseline (before randomization) were used for this analysis. SETTING: A clinic-based sample in Atlanta, Georgia. PARTICIPANTS: Eligible teens and young adults were African American female teens and young adults 15 to 21 years old who had reported sexual activity in the previous 60 days. Of 1558 teens and young adults screened from March 1, 2002, through August 31, 2004, 847 were eligible and 715 (84.4%) participated at baseline. MAIN OUTCOME MEASURES: Self-reported consistent condom use in the 14 days before baseline and Yc-PCR results. RESULTS: Of participants who reported vaginal sex in the past 14 days, 186 reported consistent condom use, defined as 100% condom use. Of these, 63 had a positive Yc-PCR result, indicating detection of the Y chromosome in the vaginal fluid. Participants who reported consistent condom use with a self-reported history of sexually transmitted diseases were 2.4 times more likely to have a positive Yc-PCR result (adjusted odds ratio, 2.4; 95% confidence interval, 1.2-4.8; P = .01). CONCLUSIONS: A significant degree of discordance between self-reports of consistent condom use and Yc-PCR positivity was observed. Several rival explanations for the observed discordance exist, including (1) teens and young adults inaccurately reported condom use; (2) teens and young adults used condoms consistently but used them incorrectly, resulting in user error; and (3) teens and young adults responded with socially desirable answers. Using an objective biological measure may provide one strategy for validating teens' and young adults' self-reported condom use. PMID- 19124706 TI - Prevalence of low back pain and its effect on health-related quality of life in adolescents. AB - OBJECTIVES: To assess the prevalence of low back pain (LBP) in adolescents and the clinical features of LBP in 2 European countries and to evaluate the effect of LBP on health-related quality of life (HRQOL) using standardized validated generic and disease-specific instruments. DESIGN: Cross-sectional study. SETTING: Secondary schools of Barcelona, Spain, and Fribourg, Switzerland. PARTICIPANTS: Representative sample of adolescents from the 2 cities. Intervention Selected adolescents completed a questionnaire including a generic HRQOL (KIDSCREEN-52) and 2 LBP-specific instruments. MAIN OUTCOME MEASURES: Results of KIDSCREEN-52, the Roland-Morris Disability Questionnaire, and the Hanover Functional Ability Questionnaire. RESULTS: A total of 1470 adolescents (52.6% male) with a mean (SD) age of 15.05 (1.17) years completed the questionnaires (response rate, 85.1%). Low back pain was reported by 587 adolescents (39.8%): isolated LBP in 250 (42.6%), LBP plus other pain in 271(46.2%), LBP plus whole-body pain in 50 (8.5%, and unclassifiable LBP in 16 (2.7%). Five hundred adolescents (34.7%) reported no pain, and 369 (25.6%) reported other pain without LBP. In those with isolated LBP, the percentage of adolescent boys was higher (54.6%; P < .001) and the LBP was mildest. In those with LBP plus whole-body pain, the percentage of adolescent girls was higher (62%; P < .001) and LBP was most severe. All KIDSCREEN scores in the group with LBP plus whole-body pain were significantly lower than in the other groups (effect size, 0.52-1.24). No differences were found between the groups who reported isolated pain, no pain, or other pain with no LBP. On the LBP specific instruments, adolescents who reported LBP plus other pain had significantly poorer scores (P < .001) compared with those with isolated LBP but better scores (P < .001) than those with LBP plus whole-body pain. CONCLUSIONS: Low back pain in adolescents is a prevalent symptom with overall low associated disability and little effect on health-related quality of life. A subset of adolescents in whom LBP is associated with whole-body pain report significant impairment and deserve more attention. PMID- 19124707 TI - Cerebral palsy, autism spectrum disorders, and developmental delay in children born after assisted conception: a systematic review and meta-analysis. AB - OBJECTIVE: To assess the existing evidence of associations between assisted conception and cerebral palsy (CP), autism spectrum disorders (ASD), and developmental delay. DATA SOURCES: Forty-one studies identified in a systematical PubMed and Excerpta Medica Database (EMBASE) search for articles published from January 1, 1996, to April 1, 2008. STUDY SELECTION: Studies written in English comparing children born after assisted conception with children born after natural conception assessing CP, ASD, and developmental delay, based on original data with a follow-up of 1 year or more. Main Exposures In vitro fertilization (IVF) with or without intracytoplasmic sperm injection or ovulation induction with or without subsequent intrauterine insemination. MAIN OUTCOME MEASURES: Cerebral palsy, ASD, and developmental delay. RESULTS: Nine CP studies showed that children born after IVF had an increased risk of CP associated with preterm delivery. In our meta-analysis including 19 462 children exposed to IVF, we estimated a crude odds ratio of 2.18 (95% confidence interval, 1.71-2.77). Eight ASD studies and 30 studies on developmental delay showed inconsistent results. No studies assessed the risk of CP, ASD, or developmental delay in children born after ovulation induction exclusively. CONCLUSIONS: Methodological problems were revealed in the identified studies, and the gaps in our knowledge about the long term outcomes of children born after assisted conception are considerable, including a lack of information on the long-term consequences of ovulation induction. Possible associations with ASD and developmental delay need assessment in larger studies. Studies on assisted conception and CP from countries outside of Scandinavia are needed, including detailed information on time to pregnancy, underlying cause of infertility, and type of IVF treatment. PMID- 19124708 TI - Picture of the month--quiz case. PMID- 19124709 TI - Social networking sites: finding a balance between their risks and benefits. PMID- 19124710 TI - Respite care: an essential yet unmet need for families with children with special health care needs. PMID- 19124711 TI - Teenagers wanting medical advice: Is MySpace the answer? PMID- 19124712 TI - Prolonged QT interval corrected for heart rate during diabetic ketoacidosis in children: psychological stress could be another explanation. PMID- 19124713 TI - Language matters: unintentional strangulation, strangulation activity, and the "choking game". PMID- 19124714 TI - Escherichia coli O157 and children. PMID- 19124715 TI - Cutting edge: Ikaros is a regulator of Th2 cell differentiation. AB - Ikaros, a hematopoietic transcription factor, has well defined effects on early lymphocyte development in the bone marrow and thymus. In this study we demonstrate that Ikaros is a positive regulator of Th2 cytokine gene expression in peripheral T cells. CD4+ T cells from naive Ikaros(null) mice cultured under Th2-skewing conditions express the Th1 cytokine IFN-gamma and have reduced IL-4, IL-5, and IL-13 expression. Ikaros directly associates with several Th2 locus regulatory regions in naive CD4+ T cells. The decreased ability to express Th2 cytokines in Ikaros(null)T cells corresponds with histone 3 hypoacetylation across the Th2 cytokine locus as well as decreased GATA3 and cMaf and increased T bet and STAT1 expression. These data support a model whereby Ikaros directly activates Th2 gene expression by promoting local chromatin accessibility during CD4+ T cell differentiation and also acts indirectly to regulate expression of Th2- and Th1-specific transcription factors. PMID- 19124716 TI - Cutting edge: developmental up-regulation of IFN-gamma-inducible lysosomal thiol reductase expression leads to reduced T cell sensitivity and less severe autoimmunity. AB - Reactivity to self-peptide/MHC complexes is required for selection of the TCR repertoire in the thymus but can also promote autoimmunity. Reduced TCR sensitivity of mature T cells is thought to help control the autoreactivity in peripheral T cells. The molecular basis for reduced sensitivity of peripheral T cells is not known. We found that peripheral T cells, but not immature thymocytes, lacking IFN-gamma-inducible lysosomal thiol reductase (GILT) display increased sensitivity to TCR ligation. GILT-/- peripheral T cells express reduced levels of mitochondrial superoxide dismutase 2 and consequently display higher levels of reactive oxygen radicals and ERK1/2 phosphorylation following activation. The increased sensitivity of GILT-deficient T cells results in a more severe hyperglycemia associated with streptozotocin-induced diabetes. GILT expression levels progressively increase in T cells with maturation. These data suggest that regulation of GILT expression may be a mechanism of T cell differentiation-associated changes in sensitivity to TCR engagement. PMID- 19124717 TI - Pre-TCR-induced beta-catenin facilitates traversal through beta-selection. AB - Pre-TCR induced signals regulate development of the alphabeta TCR lineage cells at the beta-selection checkpoint. We have previously shown that conditional deletion of beta-catenin, a central mediator of Wnt-beta-catenin-T cell factor signaling pathway, impairs traversal through the beta-selection checkpoint. We now provide a molecular basis for the impairment. We demonstrate that pre-TCR signals specifically stabilize beta-catenin in CD4-CD8- double negative thymocytes during beta-selection. Pre-TCR induced Erk activity was required to stabilize beta-catenin. Enforced expression of stabilized beta-catenin was sufficient to mediate aspects of beta-selection including sustained expression of early growth response (Egr) genes. Consistently, deletion of beta-catenin reduced induction of Egr gene expression by the pre-TCR signal and blocked efficient beta selection. Thus, we demonstrate that pre-TCR induced beta-catenin sustains expression of Egr genes that facilitate traversal through the beta-selection checkpoint. PMID- 19124718 TI - Sustained expression of pre-TCR induced beta-catenin in post-beta-selection thymocytes blocks T cell development. AB - Pre-TCR and IL-7R signals regulate beta-selection of thymocytes and then must be down-regulated for further development. However, the molecular events that control down-regulation remain unknown. We and others have previously shown that beta-catenin in cooperation with TCF regulates beta-selection. In this paper, we demonstrate that beta-catenin expression is stringently regulated by intrathymic signals, it is expressed at the highest levels in the pre-TCR signaled thymocytes, and is down-regulated in post-beta-selection thymocytes. Pre-TCR induced beta-catenin regulates initial stages of pre-TCR signaling including expression of early growth response (Egr) genes but must be down-regulated to express RORgammat, which is essential for maturation to the CD4+CD8+ double positive (DP) stage. Sustained expression of beta-catenin results in the generation of IL-7R-, Egr-, and TGFbeta-expressing pre-DP thymocytes that are blocked in development. These data are consistent with a model in which post-beta selection, pre-TCR-induced beta-catenin expression must return to background levels for efficient transition to the DP stage. PMID- 19124719 TI - Cross-linking of CD80 on CD4+ T cells activates a calcium-dependent signaling pathway. AB - CD80 expressed on the surface of APCs provides a positive costimulatory signal to naive CD4+ T cells via CD28 during activation. However, CD80 is also expressed on the surface of activated CD4+ T cells, and cross-linking CD80 on the surface of CD4+ T cells activated in the presence of Th1-promoting cytokines induces a direct up-regulation of T-bet, IFN-gamma, and Bcl(XL) expression in primary CD4+ T cells. The present data show that naive CD4+ T cells activated in Th1-promoting conditions in the presence of anti-CD80 mAb increase the level of IFN-gamma produced by increasing the rate of IFN-gamma mRNA transcription, which is supported by an increase in the level of T-bet phosphorylation and T-bet binding to the third intronic enhancer in the IFN-gamma locus. Furthermore, anti-CD80 mAb induced increase in IFN-gamma expression and T-bet phosphorylation is dependent upon the activation of a Ca2+-dependent pathway as shown by anti-CD80 mAb-induced intracellular Ca2+ flux following CD80 cross-linking. These findings indicate a novel regulatory role for CD80-mediated intracellular signals in CD4+ T cells and have important implications for disease therapies using anti-costimulatory mAbs as use of an intact CD80 mAb may lead to CD80 cross-linking on activated T cells and enhanced proinflammatory cytokine production. PMID- 19124720 TI - Priming of CD8+ and CD4+ T cells in experimental leishmaniasis is initiated by different dendritic cell subtypes. AB - The biological role of Langerin+ dendritic cells (DCs) such as Langerhans cells and a subset of dermal DCs (dDCs) in adaptive immunity against cutaneous pathogens remains enigmatic. Thus, we analyzed the impact of Langerin+ DCs in adaptive T cell-mediated immunity toward Leishmania major parasites in a Lang-DTR mouse model that allows conditional diphtheria toxin (DT)-induced ablation of Langerin+ DCs in vivo. For the first time, infection experiments with DT-treated Lang-DTR mice revealed that proliferation of L. major-specific CD8+ T cells is significantly reduced during the early phase of the immune response following depletion of Langerin+ DCs. Consequently, the total number of activated CD8+ T cells within the draining lymph node and at the site of infection is diminished. Furthermore, we show that the impaired CD8+ T cell response is due to the absence of Langerin+ dDCs and not Langerhans cells. Nevertheless, the CD4+ T cell response is not altered and the infection is cleared as effectively in DT-treated Lang-DTR mice as in control mice. This clearly demonstrates that Langerin+ DCs are, in general, dispensable for an efficient adaptive immune response against L. major parasites. Thus, we propose a novel concept that, in the experimental model of leishmaniasis, priming of CD4+ T cells is mediated by Langerin- dDCs, whereas Langerin+ dDCs are involved in early priming of CD8+ T cells. PMID- 19124722 TI - Increased CD4+Foxp3+ T cells in BAFF-transgenic mice suppress T cell effector responses. AB - The cytokine B cell activation factor of the TNF family (BAFF) is considered to perform a proinflammatory function. This paradigm is particularly true for B cell dependent immune responses; however the exact role for BAFF in regulating T cell immunity is ill-defined. To directly assess the effect of BAFF upon T cells, we analyzed T cell-dependent immune responses in BAFF-transgenic (Tg) mice. We found that T cell responses in BAFF-Tg mice are profoundly compromised, as indicated by their acceptance of islet allografts and delayed skin graft rejection. However, purified BAFF-Tg effector T cells could reject islet allografts with a normal kinetic, suggesting that the altered response did not relate to a defect in T cell function per se. Rather, we found that BAFF-Tg mice harbored an increased number of peripheral CD4+Foxp3+ T cells. A large proportion of the BAFF-expanded CD4+CD25+Foxp3+ regulatory T cells (Tregs) were CD62LlowCD103high and ICAM-1high, a phenotype consistent with an ability to home to inflammatory sites and prevent T cell effector responses. Indeed, depletion of the endogenous BAFF-Tg Tregs allowed allograft rejection to proceed, demonstrating that the increased Tregs were responsible for preventing alloimmunity. The ability of BAFF to promote Treg expansion was not T cell intrinsic, as Tregs did not express high levels of BAFF receptor 3, nor did excessive BAFF trigger NF-kappaB2 processing in Tregs. In contrast, we found that BAFF engendered Treg expansion through an indirect, B cell-dependent mechanism. Thus, under certain conditions, BAFF can play a surprising anti-inflammatory role in T cell biology by promoting the expansion of Treg cells. PMID- 19124721 TI - Clonal expansions and loss of receptor diversity in the naive CD8 T cell repertoire of aged mice. AB - There are well-characterized age-related changes in the peripheral repertoire of CD8 T cells characterized by reductions in the ratio of naive:memory T cells and the development of large clonal expansions in the memory pool. In addition, the TCR repertoire of naive T cells is reduced with aging. Because a diverse repertoire of naive T cells is essential for a vigorous response to new infections and vaccinations, there is much interest in understanding the mechanisms responsible for declining repertoire diversity. It has been proposed that one reason for declining repertoire diversity in the naive T cell pool is an increasing dependence on homeostatic proliferation in the absence of new thymic emigrants for maintenance of the naive peripheral pool. In this study, we have analyzed the naive CD8 T cell repertoire in young and aged mice by DNA spectratype and sequence analysis. Our data show that naive T cells from aged mice have perturbed spectratype profiles compared with the normally Gaussian spectratype profiles characteristic of naive CD8 T cells from young mice. In addition, DNA sequence analysis formally demonstrated a loss of diversity associated with skewed spectratype profiles. Unexpectedly, we found multiple repeats of the same sequence in naive T cells from aged but not young mice, consistent with clonal expansions previously described only in the memory T cell pool. Clonal expansions among naive T cells suggests dysregulation in the normal homeostatic proliferative mechanisms that operate in young mice to maintain diversity in the naive T cell repertoire. PMID- 19124723 TI - IL-20 receptor 2 signaling down-regulates antigen-specific T cell responses. AB - The recently described cytokines IL-19, IL-20, and IL-24 share structural homology with IL-10 and are therefore classified as members of the IL-10 family of cytokines. Although it has long been speculated that signaling by their heterodimeric receptor complexes (IL-20R1/IL-20R2 and IL-22R/IL-20R2) influences immunological processes, the target cells for this group of cytokines are still unclear. By generating a knockout mouse strain deficient for the common IL-20R beta-chain (IL-20R2), we show that IFN-gamma and IL-2 secretion is significantly elevated after stimulation of IL-20R2-/--deficient CD8 and CD4 T cells with Con A or anti-CD3/CD28 in vitro. IL-10 secretion by activated IL-20R2-/- CD4 cells was diminished. Consistent with our in vitro results, significantly more Ag-specific CD8 IFN-gamma+ and CD4 IFN-gamma+ T cells developed to locally applied DNA vaccines in IL-20R2-deficient mice. In a T cell-dependent model of contact hypersensitivity, IL-20R2 knockout mice were more sensitive to the contact allergen trinitro-chloro-benzene. Thus, IL-20R2 signaling directly regulates CD8 and CD4 T cell answers in vitro and in vivo. For the first time, we provide evidence that IL-19, IL-20, and IL-24 are part of a signaling network that normally down-modulates T cell responses in mice. PMID- 19124724 TI - Tuberculosis-induced variant IL-4 mRNA encodes a cytokine functioning as growth factor for (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate-specific Vgamma2Vdelta2 T cells. AB - The possibility that mycobacterial infections induce variant cytokine mRNA encoding a functionally distinct protein for immune regulation has not been addressed. In this study, we reported that Mycobacterium tuberculosis and bacillus Calmette-Guerin infections of macaques induced expression of variant IL 4 (VIL-4) mRNA encoding a protein comprised of N-terminal 97 aa identical with IL 4, and unique C-terminal 96 aa including a signaling-related proline-rich motif. While VIL-4 could be stably produced as intact protein, the purified VIL-4 induced apparent expansion of phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP)-specific Vgamma2Vdelta2 T cells in dose- and time-dependent manners. The unique C-terminal 96 aa bearing the proline-rich motif (PPPCPP) of VIL-4 appeared to confer the ability to expand Vgamma2Vdelta2 T cells, since simultaneously produced IL-4 had only a subtle effect on these gammadelta T cells. Moreover, VIL-4 seemed to use IL-4R alpha for signaling and activation, as the VIL-4-induced expansion of Vgamma2Vdelta2 T cells was blocked by anti-IL-4R alpha mAb but not anti-IL-4 mAb. Surprisingly, VIL-4-expanded Vgamma2Vdelta2 T cells after HMBPP stimulation appeared to be heterologous effector cells capable of producing IL-4, IFN-gamma, and TNF-alpha. Thus, mycobacterial infections of macaques induced variant mRNA encoding VIL-4 that functions as growth factor promoting expansion of HMBPP-specific Vgamma2Vdelta2 T effector cells. PMID- 19124725 TI - TLR4 ligands induce IFN-alpha production by mouse conventional dendritic cells and human monocytes after IFN-beta priming. AB - Exacerbation of disease in systemic lupus erythematosus (SLE) is associated with bacterial infection. In conventional dendritic cells (cDCs), the TLR4 ligand bacterial LPS induces IFN-beta gene expression but does not induce IFN-alpha. We hypothesized that when cDCs are primed by cytokines, as may frequently be the case in SLE, LPS would then induce the production of IFN-alpha, a cytokine believed to be important in lupus pathogenesis. In this study we show that mouse cDCs and human monocytes produce abundant IFN-alpha following TLR4 engagement whether the cells have been pretreated either with IFN-beta or with a supernatant from DCs activated by RNA-containing immune complexes from lupus patients. This TLR4-induced IFN-alpha induction is mediated by both an initial TRIF-dependent pathway and a subsequent MyD88-dependent pathway, in contrast to TLR3-induced IFN alpha production, which is entirely TRIF-dependent. There is also a distinct requirement for IFN regulatory factors (IRFs), with LPS-induced IFN-alpha induction being entirely IRF7- and partially IRF5-dependent, in contrast to LPS induced IFN-beta gene induction which is known to be IRF3-dependent but largely IRF7-independent. This data demonstrates a novel pathway for IFN-alpha production by cDCs and provides one possible explanation for how bacterial infection might precipitate disease flares in SLE. PMID- 19124726 TI - IL-12-dependent inducible expression of the CD94/NKG2A inhibitory receptor regulates CD94/NKG2C+ NK cell function. AB - The inhibitory CD94/NKG2A and activating CD94/NKG2C killer lectin-like receptors specific for HLA-E have been reported to be selectively expressed by discrete NK and T cell subsets. In the present study, minor proportions of NK and T cells coexpressing both CD94/NKG2A and CD94/NKG2C were found in fresh peripheral blood from adult blood donors. Moreover, CD94/NKG2A surface expression was transiently detected upon in vitro stimulation of CD94/NKG2C+ NK cells in the presence of irradiated allogeneic PBMC or rIL-12. A similar effect was observed upon coculture of NKG2C+ NK clones with human CMV-infected autologous dendritic cell cultures, and it was prevented by an anti-IL-12 mAb. NKG2A inhibited the cytolytic activity of NKG2C+ NK clones upon engagement either by a specific mAb or upon interaction with a transfectant of the HLA class I-deficient 721.221 cell line expressing HLA-E. These data indicate that beyond its constitutive expression by an NK cell subset, NKG2A may be also transiently displayed by CD94/NKG2C+ NK cells under the influence of IL-12, providing a potential negative regulatory feedback mechanism. PMID- 19124727 TI - Epicutaneous immunization with protein antigen in the presence of TLR4 ligand induces TCR alpha beta+CD4+ T contrasuppressor cells that reverse skin-induced suppression of Th1-mediated contact sensitivity. AB - Our previous work showed that epicutaneous (EC) immunization of mice with different protein Ags applied on the skin in the form of a patch induces a state of subsequent Ag-nonspecific unresponsiveness due to suppressor CD4+8+ T cells (Ts) that inhibit Th1-mediated contact sensitivity (CS) reactions via released TGF-beta. In the present work we show that EC immunization with Ag together with the TLR4 ligand LPS induced cells that could prevent suppression by the Ag nonspecific Ts. These up-regulatory cells, called contrasuppressor T cells (Tcs), belong to a population of Ag-specific TCRalphabeta CD4+ lymphocytes and are different from Th1 CD4+ cells that mediate the CS reaction. Experiments using knockout mice showed that EC induced contrasuppression is MyD88, INF-gamma, and IL-12 dependent, whereas IL-6 is not involved in this phenomenon. Additional experiments with anti-IFN-gamma mAb showed that IFN-gamma is required for induction of Tcs cells but does not play a crucial role in the effector phase of contrasuppression. Additionally, treatment of CS effector cells with rIL-12 makes them resistant to EC induced suppression without affecting Ts cells, whereas IL 12 neutralization in vitro abrogates contrasuppression. These data show that IL 12 is indeed involved in the effector phase of EC induced contrasuppression and that this cytokine does not act directly on Ts cells. The mechanism of action of Tcs protects Th1 effector cells mediating CS from the nonspecific Ts, leaving suppression to other Ags intact. Ts and Tcs cells do not influence each other and can be induced simultaneously in the same animal. PMID- 19124728 TI - The EBV-encoded dUTPase activates NF-kappa B through the TLR2 and MyD88-dependent signaling pathway. AB - The innate immune response plays a key role as the primary host defense against invading pathogens including viruses. We have previously shown that treatment of human monocyte-derived macrophages with EBV-encoded dUTPase induces the expression of proinflammatory cytokines through the activation of NF-kappaB. However, the receptor responsible for EBV-encoded dUTPase-mediated biological effects is not known. In this study, we demonstrate that the purified EBV-encoded dUTPase activates NF-kappaB in a dose-dependent manner through TLR2 and requires the recruitment of the adaptor molecule MyD88 but not CD14. Furthermore, activation of NF-kappaB was abrogated by anti-TLR2, anti-EBV-encoded dUTPase blocking Abs and the overexpression of a dominant negative construct of MyD88 in human embryonic kidney 293 cells expressing TLR2. In addition, treatment of human monocyte-derived macrophages with the anti-EBV-encoded dUTPase Ab 7D6 or the anti TLR2 Ab blocked the production of IL-6 by the EBV-encoded dUTPase. To our knowledge, this is the first report demonstrating that a nonstructural protein encoded by EBV is a pathogen-associated molecular pattern and that it has immunomodulatory functions. Although additional studies are necessary to define the signaling pathways activated by the EBV-encoded dUTPase and to determine its role in modulating immune responses to EBV infection, our results suggest that the dUTPase could be a potential target for the development of novel therapeutic agents against infections caused by EBV. PMID- 19124729 TI - Attenuated expression of A20 markedly increases the efficacy of double-stranded RNA-activated dendritic cells as an anti-cancer vaccine. AB - A20 is a zinc finger protein with ubiquitin-modifying activity. A20 has been described as negatively regulating signaling induced by the TNF receptor and TLR family in a number of cell types, including mouse bone marrow-derived dendritic cells (DCs). However, the expression and effect of A20 in activated human monocyte-derived DCs have not been previously evaluated. We report that DCs activated with the TLR3 ligand poly(I:C) up-regulate A20. Down-regulating A20 demonstrated its role in the functional activation of DCs. A20 down-regulated DCs showed higher activation of the transcription factors NF-kappaB and activator protein-1, which resulted in increased and sustained production of IL-6, IL-10, and IL-12p70. We additionally silenced the immunosuppressive cytokine IL-10 and demonstrated that IL-10 inhibits T cell proliferation. We further demonstrated that A20 down-regulated DCs skew naive CD4+ T cells toward IFN-gamma producing Th1 cells, a process which is dependent on IL-12p70 and which is unaffected by IL 10. Furthermore, A20 and/or IL-10 down-regulated DCs had an enhanced capacity to prime Melan-A/MART-1 specific CD8+ T cells. Finally, we demonstrated that potent T cell stimulatory DCs are generated by the simultaneous delivery of poly(I:C12U), A20, or A20/IL-10 small interfering RNA and Ag-encoding mRNA, introducing a one step approach to improve DC-based vaccines. Together these findings demonstrate that A20 negatively regulates NF-kappaB and activator protein-1 in DCs and that down-regulation of A20 results in DCs with enhanced T cell stimulatory capacity. PMID- 19124730 TI - Plasmacytoid dendritic cells are dispensable during primary influenza virus infection. AB - Plasmacytoid dendritic cells (pDC) are thought to be pivotal in the first line of defense against viral infections. Although previous studies have suggested that pDC regulate the immune response against respiratory syncytial virus, their role in pulmonary infection with influenza virus has remained unclear. Using mice with GFP-tagged pDC, we observed a marked increase in pDC numbers in the lung airways 3 days after intranasal infection with influenza virus A/PR/8/34. To further investigate their potential involvement in the disease, we made use of pDC deficient IkarosL/L mice. In the absence of pDC, the recruitment of T cells to the bronchoalveolar space was delayed, which could be reversed by the adoptive transfer of pDC before infection. Surprisingly, however, when compared with wild type animals, IkarosL/L mice revealed a similar course of disease, as determined by weight loss, viral titers, levels of neutralizing Ab, and lung pathology. Moreover, the activation and differentiation of influenza-specific CD8+ effector T cells was unaltered in the absence of pDC, as was the generation of CD8+ memory T cells. Taken together, our study suggests that pDC regulate the accumulation of T cells in the bronchoalveolar space during early influenza virus infection, but are dispensable for the control of this disease. PMID- 19124731 TI - Modified adenine (9-benzyl-2-butoxy-8-hydroxyadenine) redirects Th2-mediated murine lung inflammation by triggering TLR7. AB - Substitute adenine (SA)-2, a synthetic heterocycle chemically related to adenine with substitutions in positions 9-, 2-, and 8- (i.e., 9-benzyl-2-butoxy-8 hydroxyadenine), induces in vitro immunodeviation of Th2 cells to a Th0/Th1 phenotype. In this article, we evaluate the in vivo ability of SA-2 to affect Th2 mediated lung inflammation and its safety. TLR triggering and NF-kappaB activation by SA-2 were analyzed on TLR-transfected HEK293 cells and on purified bone marrow dendritic cells. The in vivo effect of SA-2 on experimental airway inflammation was evaluated in both prepriming and prechallenge protocols by analyzing lung inflammation, including tissue eosinophilia and goblet cell hyperplasia, bronchoalveolar lavage fluid cell types, and the functional profile of Ag-specific T cells from draining lymph nodes and spleens. SA-2 induced mRNA expression and production of proinflammatory (IL-6, IL-12, and IL-27) and regulatory (IL-10) cytokines and chemokines (CXCL10) in dendritic cells but down regulated TGF-beta. Prepriming administration of SA-2 inhibited OVA-specific Abs and Th2-driven lung inflammation, including tissue eosinophilia and goblet cells, with a prevalent Foxp3-independent regulatory mechanism. Prechallenge treatment with SA-2 reduced the lung inflammation through the induction of a prevalent Th1 related mechanism. In this model the activity of SA-2 was route-independent, but adjuvant- and Ag dose-dependent. SA-2-treated mice did not develop any increase of serum antinuclear autoantibodies. In conclusion, critical substitutions in the adenine backbone creates a novel synthetic TLR7 ligand that shows the ability to ameliorate Th2-mediated airway inflammation by a complex mechanism, involving Th1 redirection and cytokine-mediated regulation, which prevents autoreactivity. PMID- 19124732 TI - Resting human memory B cells are intrinsically programmed for enhanced survival and responsiveness to diverse stimuli compared to naive B cells. AB - Enhanced secondary Ab responses are a vital component of adaptive immunity, yet little is understood about the intrinsic and extrinsic regulators of naive and memory B cells that result in differences in their responses to Ag. Microarray analysis, together with surface and intracellular phenotyping, revealed that memory B cells have increased expression of members of the TNF receptor, SLAM (signaling lymphocytic activation molecule), B7, and Bcl2 families, as well as the TLR-related molecule CD180 (RP105). Accordingly, memory B cells exhibited enhanced survival, proliferation, and Ig secretion, and they entered division more rapidly than did naive B cells in response to both T cell-dependent and T cell-independent stimuli. Furthermore, both IgM and isotype-switched memory B cells, but not naive B cells, costimulated CD4+ T cells in vitro through a mechanism dependent on their constitutive expression of CD80 and CD86. This study demonstrates that up-regulation of genes involved in activation, costimulation, and survival provides memory B cells with a unique ability to produce enhanced immune responses and contributes to the maintenance of the memory B cell pool. PMID- 19124733 TI - Kinetics of major histocompatibility class I antigen presentation in acute infection. AB - Ag presentation within the regional lymph node is crucial for the initiation of CD8+ T cell responses following viral infection. The magnitude and quality of the CD8+ T cell response are regulated by the interplay between the size of the APC population and duration of Ag presentation. To understand how these parameters are finely regulated during an immune response, we have investigated the dynamics of Ag presentation in influenza A virus and HSV-1 infection. In both infections, APC production was calculated to occur over the first few days of infection, after which there was slow exponential decay over a period of up to 2 wk. This production rate is most likely determined by the Ag availability and recruitment and/or maturation rate of dendritic cells. APC production was found to closely parallel lymph node cell recruitment in both infections. This was greatest in the first 6 h of infection for HSV and over the second and third day for influenza. In HSV infection, the peak production also coincides with peak viral levels. By contrast, in influenza infection, APC production ceased between the third and fourth day despite the presence of high levels of virus until 5 days after infection. These analyses demonstrate that two quite different self-limiting infections generate the APC necessary to drive T cell responses early in infection at different rates. Understanding how such contrasting kinetics of Ag presentation impacts on the growth and size of developing protective T cell populations has important implications for the design of vaccines and immunotherapies. PMID- 19124734 TI - Graft-versus-host disease prevents the maturation of plasmacytoid dendritic cells. AB - The role of Ag presenting cell subsets in graft-versus-host disease (GVHD) remains unclear. We have thus examined the ability of plasmacytoid dendritic cells (pDC) to modulate transplant outcome. Surprisingly, host pDC were exquisitely sensitive to total body irradiation and were depleted before transplantation, thus allowing us to focus on donor pDC [corrected]. The depletion of all pDC from bone marrow grafts resulted in an acceleration of GVHD mortality while the depletion of mature pDC from G-CSF mobilized splenic grafts had no effect. Thus, donor bone marrow pDC, but not mature pDC contained within stem cell grafts attenuate acute GVHD. In the presence of GVHD, donor pDC completely failed to reconstitute although a CD11clow120G8+ precursor DC reconstituted in an exaggerated and transient manner. These cells expressed Flt 3, the macrophage colony stimulating factor receptor and, consistent with a common dendritic cell (DC) precursor, were capable of differentiation into pDC and conventional DC in vivo in the absence of GVHD. These precursors were MHC class II+ and CD80/86+ but lacked CD40, were actively presenting host Ag and inhibited GVHD and T cell proliferation in a contact-dependent fashion. These data demonstrate that GVHD prevents the maturation of pDC and instead promotes the generation of a suppressive precursor DC, further contributing to the state of immune paralysis after transplantation. PMID- 19124735 TI - Differential capability of human cutaneous dendritic cell subsets to initiate Th17 responses. AB - Human skin-migratory dendritic cells (DCs) have the ability to prime and bias Th1 and Th2 CD4+ T lymphocytes. However, whether human cutaneous DCs are capable of initiating proinflammatory Th17 responses remains undetermined. We report that skin-migratory DCs stimulate allogeneic naive CD4+ T cells that differentiate simultaneously into two distinct effector Th17 and Th1 populations capable of homing to the skin, where they induce severe cutaneous damage. Skin-migratory Langerhans cells (smiLCs) were the main cutaneous DC subset capable of inducing Th17 responses dependent on the combined effects of IL-15 and stabilized IL-6, which resulted in IL-6 trans-signaling of naive CD4+ T cells. Different from smiLCs, purified skin-migratory dermal DCs did not synthesize IL-15 and were unable to bias Th17 responses. Nevertheless, these dermal DCs were capable of differentiating Th17 cells in mixed leukocyte cultures supplemented with IL-15 and stabilized IL-6. Overall, our data demonstrate that human epidermal smiLCs induce Th17 responses by mechanisms different from those previously described and highlight the need to target clinical treatments based on these variations. PMID- 19124736 TI - Endogenous 4-1BB ligand plays a critical role in protection from influenza induced disease. AB - A critical issue during severe respiratory infection is whether it is the virus or the host response that does the most damage. In this study, we show that endogenous 4-1BBL plays a critical role in protecting mice from severe effects of influenza disease. During mild respiratory influenza infection in which virus is rapidly cleared, the inducible costimulatory receptor 4-1BB is only transiently induced on lung T cells and 4-1BB ligand (4-1BBL) is completely dispensable for the initial CD8 T cell response and mouse survival. In contrast, during more severe respiratory influenza infection with prolonged viral load, 4-1BB expression on lung CD8 T cells is sustained, and 4-1BBL-deficient mice show decreased CD8 T cell accumulation in the lungs, decreased viral clearance, impaired lung function, and increased mortality. Transfer of an optimal number of naive Ag-specific T cells before infection protects wild-type but not 4-1BBL deficient mice from an otherwise lethal dose of influenza virus. Transfer of T cells lacking the proapoptotic molecule Bim extends the lifespan of 4-1BBL deficient mice by one to three days, suggesting that at least part of the role of 4-1BB/4-1BBL is to prolong effector cell survival long enough to clear virus. Intranasal delivery of 4-1BBL by recombinant adenovirus marginally improves survival of 4-1BBL-deficient mice at low dose, but exacerbates disease at high dose. These findings suggest a rationale for the evolutionary accumulation of inducible costimulatory molecules, thereby allowing the immune system to sustain the expression of molecules such as 4-1BB to a level commensurate with severity of infection. PMID- 19124737 TI - A dual role for talin in NK cell cytotoxicity: activation of LFA-1-mediated cell adhesion and polarization of NK cells. AB - LFA-1 is critical for NK cell cytotoxicity because it mediates adhesion of NK cells to target cells. Talin is thought to associate with the cytoplasmic tail of LFA-1 and activates its ligand-binding function. In this study, we report that talin is also required for LFA-1-mediated outside-in signaling leading to NK cell polarization. NK cells generated from talin1-deficient murine embryonic stem cells are defective in LFA-1-mediated adhesion. Although exogenously added manganese activates LFA-1 on talin-deficient NK cells and induces conjugate formation with target cells, their LFA-1-dependent cytotoxicity is impaired. Binding of ICAM-1-coated beads to wild-type NK cells induces reorganization of the actin cytoskeleton and coligation of the activating receptor NKG2D induces polarization of cytotoxic granules, whereas talin1-deficient NK cells fail to polarize with or without NKG2D coligation. Thus, talin1 plays a dual role in NK cell cytotoxicity, first by activation of LFA-1-mediated adhesion and then via LFA-1-induced NK cell polarization. PMID- 19124738 TI - RhoH plays critical roles in Fc epsilon RI-dependent signal transduction in mast cells. AB - RhoH is an atypical small G protein with defective GTPase activity that is specifically expressed in hematopoietic lineage cells. RhoH has been implicated in regulation of several physiological processes including hematopoiesis, integrin activation, and T cell differentiation and activation. In the present study, we investigated the role of RhoH in mast cells by generating RhoH knockout mice. Despite observing normal development of mast cells in vivo, passive systemic anaphylaxis and histamine release were impaired in these mice. We also observed defective degranulation and cytokine production upon FcepsilonRI ligation in RhoH-deficient bone marrow-derived mast cells. Furthermore, FcepsilonRI-dependent activation of Syk and phosphorylation of its downstream targets, including LAT, SLP76, PLCgamma1, and PLCgamma2 were impaired, however phosphorylation of the gamma-subunit of FcepsilonRI remained intact. We also found RhoH-Syk association that was greatly enhanced by active Fyn. Our results indicate that RhoH regulates FcepsilonRI signaling in mast cells by facilitating Syk activation, possibly as an adaptor molecule for Syk. PMID- 19124739 TI - Plasmacytoid dendritic cells regulate breach of self-tolerance in autoimmune arthritis. AB - Achieving remission in rheumatoid arthritis (RA) remains elusive despite current biological therapeutics. Consequently, interest has increased in strategies to re establish immune tolerance to provide long-term disease suppression. Although dendritic cells (DC) are prime candidates in initiating autoreactive T cell responses, and their presence within the synovial environment suggests a role in generation and maintenance of autoreactive, synovial T cell responses, their functional importance remains unclear. We investigated the contribution made by plasmacytoid DCs (pDCs) in the spontaneous breach of tolerance to arthritis related self proteins, including rheumatoid factor, citrullinated peptide, and type II collagen observed in a novel arthritis model. Selective pDC depletion in vivo enhanced the severity of articular pathology and enhanced T and B cell autoimmune responses against type II collagen. pDC may offer a net anti inflammatory function in the context of articular breach of tolerance. Such data will be vital in informing DC modulatory/therapeutic approaches. PMID- 19124740 TI - Lymphopenia-induced proliferation is a potent activator for CD4+ T cell-mediated autoimmune disease in the retina. AB - To study retinal immunity in a defined system, a CD4+ TCR transgenic mouse line (betagalTCR) specific for beta-galactosidase (betagal) was created and used with transgenic mice that expressed betagal in retinal photoreceptor cells (arrbetagal mice). Adoptive transfer of resting betagalTCR T cells, whether naive or Ag experienced, into arrbetagal mice did not induce retinal autoimmune disease (experimental autoimmune uveoretinitis, EAU) and gave no evidence of Ag recognition. Generation of betagalTCR T cells in arrbetagal mice by use of bone marrow grafts, or double-transgenic mice, also gave no retinal disease or signs of Ag recognition. Arrbetagal mice were also resistant to EAU induction by adoptive transfer of in vitro-activated betagalTCR T cells, even though the T cells were pathogenic if the betagal was expressed elsewhere. In vitro manipulations to increase T cell pathogenicity before transfer did not result in EAU. The only strategy that induced a high frequency of severe EAU was transfer of naive, CD25-depleted, betagalTCR T cells into lymphopenic arrbetagal recipients, implicating regulatory T cells in the T cell inoculum, as well as in the recipients, in the resistance to EAU. Surprisingly, activation of the CD25 depleted betagalTCR T cells before transfer into the lymphopenic recipients reduced EAU. Taken together, the results suggest that endogenous regulatory mechanisms, as well as peripheral induction of regulatory T cells, play a role in the protection from EAU. PMID- 19124741 TI - PD-1 blockade in rhesus macaques: impact on chronic infection and prophylactic vaccination. AB - Programmed Cell Death 1 (PD-1) plays a crucial role in immunomodulation. Binding of PD-1 to its ligand receptors down-regulates immune responses, and published reports suggest that this immune modulation is exploited in cases of tumor progression or chronic viral infection to evade immune surveillance. Thus, blockade of this signal could restore or enhance host immune functions. To test this hypothesis, we generated a panel of mAbs specific to human PD-1 that block PD ligand 1 and tested them for in vitro binding, blocking, and functional T cell responses, and evaluated a lead candidate in two in vivo rhesus macaque (Macaca mulatta) models. In the first therapeutic model, chronically SIV-infected macaques were treated with a single infusion of anti-PD-1 mAb; viral loads increased transiently before returning to, or falling below, pretreatment baselines. In the second prophylactic model, naive macaques were immunized with an SIV-gag adenovirus vector vaccine. Induced PD-1 blockade caused a statistically significant (p<0.05) increase in the peak percentage of T cells specific for the CM9 Gag epitope. These new results on PD-1 blockade in nonhuman primates point to a broader role for PD-1 immunomodulation and to potential applications in humans. PMID- 19124742 TI - Generation of a novel system for studying spleen tyrosine kinase function in macrophages and B cells. AB - Spleen tyrosine kinase (Syk) is a nonreceptor tyrosine kinase that is expressed primarily in hematopoietic cells. Because this protein has been implicated in processes such as Fc-mediated phagocytosis, BCR signaling, oxidative burst, degranulation, cytokine secretion, and integrin-mediated outside-in signaling, it is hypothesized that Syk may be a viable target in the treatment of a variety of autoimmune and inflammatory diseases. Because efforts to design a small-molecule therapeutic that specifically inhibits Syk have been largely unsuccessful, and genetic studies of Syk have been hampered by the fact that syk-/- mice die in utero, we have taken a chemical genetic approach to study the function of Syk. Specifically, we have created a mutant form of Syk that retains its wild-type function, but is susceptible to inhibition by enlarged derivatives of the tyrosine kinase inhibitor, PP1. We report in this study that Syk M442A S505A reconstituted wild-type function when introduced into murine syk-/- bone marrow derived macrophages and syk-/- DT40 chicken B cells, as determined by functional and biochemical assays. Furthermore, after screening a series of PP1 derivatives, we identified one compound, namely 2,3-DMB-PP1, that specifically inhibited Syk M442A S505A, but not wild-type Syk. This system provides us with the power to characterize immune functions that are Syk specific, and furthermore, it provides us with a tool to assess how inhibition of Syk may alter an immune response and influence disease pathogenesis and/or progression. PMID- 19124743 TI - Critical role for the transcription regulator CCCTC-binding factor in the control of Th2 cytokine expression. AB - Differentiation of naive CD4+ cells into Th2 cells is accompanied by chromatin remodeling at the Th2 cytokine locus allowing the expression of the IL-4, IL-5, and IL-13 genes. In this report, we investigated the role in Th2 differentiation of the transcription regulator CCCTC-binding factor (CTCF). Chromatin immunoprecipitation analysis revealed multiple CTCF binding sites in the Th2 cytokine locus. Conditional deletion of the Ctcf gene in double-positive thymocytes allowed development of peripheral T cells, but their activation and proliferation upon anti-CD3/anti-CD28 stimulation in vitro was severely impaired. Nevertheless, when TCR signaling was circumvented with phorbol ester and ionomycin, we observed proliferation of CTCF-deficient T cells, enabling the analysis of Th2 differentiation in vitro. We found that in CTCF-deficient Th2 polarization cultures, transcription of IL-4, IL-5, and IL-13 was strongly reduced. By contrast, CTCF deficiency had a moderate effect on IFN-gamma production in Th1 cultures and IL-17 production in Th17 cultures was unaffected. Consistent with a Th2 cytokine defect, CTCF-deficient mice had very low levels of IgG1 and IgE in their serum, but IgG2c was close to normal. In CTCF-deficient Th2 cultures, cells were polarized toward the Th2 lineage, as substantiated by induction of the key transcriptional regulators GATA3 and special AT-rich binding protein 1 (SATB1) and down-regulation of T-bet. Also, STAT4 expression was low, indicating that in the absence of CTCF, GATA3 still operated as a negative regulator of STAT4. Taken together, these findings show that CTCF is essential for GATA3- and SATB1-dependent regulation of Th2 cytokine gene expression. PMID- 19124744 TI - JNK MAPK pathway regulates constitutive transcription of CCL5 by human NK cells through SP1. AB - The MAPKs ERK, JNK, and p38 control diverse aspects of the immune response, including regulation of cytotoxin biology in NK cells and CTL. The chemokine CCL5 is coreleased with the cytotoxins, perforin, the granzymes, and granulysin, during the lethal hit administered by cytotoxic CD8+ T cells (CTL). CCL5 expression is up-regulated relatively late in CTL coincident with their functional maturation 3-7 days after activation. Unlike T cells, NK cells have the ability to kill virally infected or transformed cells when directly isolated from the peripheral circulation. In this study, we show that in contrast to T cells, peripheral blood NK cells express CCL5 constitutively. The use of specific inhibitors of the JNK, ERK, and p38 MAPK pathways showed that the JNK pathway controls expression of CCL5 by NK cells. Promoter-reporter assays identified a compact region of the CCL5 promoter responsible for the constitutive transcription of CCL5 by NK cells. EMSA, chromatin immune precipitation, the use of heterologous promoters, and site-directed mutagenesis demonstrated that transcription in NK cells is largely controlled through binding of the transcription factor specificity protein 1 to a region -75 to -56 upstream of the site of transcriptional initiation. Specificity protein 1 expression, and in turn the constitutive expression of CCL5, was found to be controlled through constitutive activation of the JNK/MAPK pathway in peripheral blood NK cells. PMID- 19124745 TI - Ezrin and moesin function together to promote T cell activation. AB - The highly homologous proteins ezrin, radixin, and moesin link proteins to the actin cytoskeleton. The two family members expressed in T cells, ezrin and moesin, are implicated in promoting T cell activation and polarity. To elucidate the contributions of ezrin and moesin, we conducted a systematic analysis of their function during T cell activation. In response to TCR engagement, ezrin and moesin were phosphorylated in parallel at the regulatory threonine, and both proteins ultimately localized to the distal pole complex (DPC). However, ezrin exhibited unique behaviors, including tyrosine phosphorylation and transient localization to the immunological synapse before movement to the DPC. To ask whether these differences reflect unique requirements for ezrin vs moesin in T cell signaling, we generated mice with conditional deletion of ezrin in mature T cells. Ezrin-/- T cells exhibited normal immunological synapse organization based upon localization of protein kinase C-theta, talin, and phospho-ZAP70. DPC localization of CD43 and RhoGDP dissociation inhibitor, as well as the novel DPC protein Src homology region 2 domain-containing phosphatase-1, was also unaffected. However, recruitment of three novel DPC proteins, ezrin binding protein of 50 kDa, Csk binding protein, and the p85 subunit of PI3K was partially perturbed. Biochemical analysis of ezrin-/- T cells or T cells suppressed for moesin using small interfering RNA showed intact early TCR signaling, but diminished levels of IL-2. The defects in IL-2 production were more pronounced in T cells deficient for both ezrin and moesin. These cells also exhibited diminished phospholipase C-gamma1 phosphorylation and calcium flux. We conclude that despite their unique movement and phosphorylation patterns, ezrin and moesin function together to promote T cell activation. PMID- 19124746 TI - Ig-like transcript 4 inhibits lipid antigen presentation through direct CD1d interaction. AB - NKT cells recognize lipid Ags presented by CD1d molecules and play an important role in the regulation of innate and adaptive immune responses. In this study, we report the identification of a membrane-associated protein, Ig-like transcript 4 (ILT4), as a novel human CD1d receptor that inhibits CD1d-mediated immune responses. We found that native CD1d tetramer generated by mammalian cells was able to specifically bind human monocytes in the peripheral blood, and this binding was at least partly mediated by monocyte-expressed ILT4. The interaction between ILT4 and CD1d involves the two N-terminal domains of ILT4 and the Ag binding groove of CD1d (alpha1/alpha2 domain). This interaction has been identified on the cell surface as well as in the endosomal and lysosomal compartments. Functional analysis showed that ILT4 could block the loading of lipid Ags such as alpha-GalCer, and consequently inhibited NKT recognition. The interaction between ILT4 and CD1d may provide new insights into the regulation of NKT-mediated immunity. PMID- 19124747 TI - Regulation of the foxp3 gene by the Th1 cytokines: the role of IL-27-induced STAT1. AB - Impaired functional activity of T regulatory cells has been reported in allergic patients and results in an increased susceptibility to autoimmune diseases. The master regulator of T regulatory cell differentiation, the transcription factor FOXP3, is required for both their development and function. Despite its key role, relatively little is known about the molecular mechanisms regulating foxp3 gene expression. In the present study, the effect of Th1 cytokines on human T regulatory cell differentiation was analyzed at epigenetic and gene expression levels and reveals a mechanism by which the STAT1-activating cytokines IL-27 and IFN-gamma amplify TGF-beta-induced FOXP3 expression. This study shows STAT1 binding elements within the proximal part of the human FOXP3 promoter, which we previously hypothesized to function as a key regulatory unit. Direct binding of STAT1 to the FOXP3 promoter following IL-27 stimulation increases its transactivation process and induces permissive histone modifications in this key region of the FOXP3 promoter, suggesting that FOXP3 expression is promoted by IL 27 by two mechanisms. Our data demonstrate a molecular mechanism regulating FOXP3 expression, which is of considerable interest for the development of new drug targets aiming to support anti-inflammatory mechanisms of the immune system. PMID- 19124748 TI - 14-3-3 gamma is stimulated by IL-3 and promotes cell proliferation. AB - IL-3 plays important roles in the growth and survival of hematopoietic progenitor cells, processes modeled in studies of the IL-3-dependent cell line Ba/F3. To gain insights into molecular mechanisms governing cell fate, we examined the patterns of proteins up-regulated following stimulation of Ba/F3 cells with IL-3. Through two-dimensional electrophoresis and proteomics-based approaches, we identified 11 proteins. Of these, expression of 14-3-3gamma was significantly increased by IL-3 stimulation at both the transcriptional and translational levels. 14-3-3gamma overexpression in Ba/F3 cells abrogated dependence on IL-3 and was associated with activation of PI3K and MAPK signaling cascades, suggesting that the functions of 14-3-3gamma in normal hematopoietic progenitors are to promote survival and growth through the activation of distinct signaling pathways. Additionally, the up-regulation of Bax and Bad was seen with the ablation of 14-3-3gamma, resulting in cell death. These results indicate that deregulated expression of 14-3-3gamma may contribute to malignant transformation, possibly providing a new target for therapeutic intervention in hematopoietic neoplasms. PMID- 19124749 TI - Polyanion-induced self-association of complement factor H. AB - Factor H is the primary soluble regulator of activation of the alternative pathway of complement. It prevents activation of complement on host cells and tissues upon association with C3b and surface polyanions such as sialic acids, heparin, and other glycosaminoglycans. Here we show that interaction with polyanions causes self-association forming tetramers of the 155,000 Da glycosylated protein. Monomeric human factor H is an extended flexible protein that exhibits an apparent size of 330,000 Da, relative to globular standards, during gel filtration chromatography in the absence of polyanions. In the presence of dextran sulfate (5000 Da) or heparin an intermediate species of apparent m.w. 700,000 and a limit species of m.w. 1,400,000 were observed by gel filtration. Sedimentation equilibrium analysis by analytical ultracentrifugation indicated a monomer Mr of 163,000 in the absence of polyanions and a Mr of 607,000, corresponding to a tetramer, in the presence of less than a 2-fold molar excess of dextran sulfate. Increasing concentrations of dextran sulfate increased binding of factor H to zymosan-C3b 4.5-fold. This result was accompanied by an increase in both the decay accelerating and cofactor activity of factor H on these cells. An expressed fragment encompassing the C-terminal polyanion binding site (complement control protein domains 18-20) also exhibited polyanion-induced self-association, suggesting that the C-terminal ends of factor H mediate self association. The results suggest that recognition of polyanionic markers on host cells and tissues by factor H, and the resulting regulation of complement activation, may involve formation of dimers and tetramers of factor H. PMID- 19124750 TI - Cell-mediated immunity to Toxoplasma gondii develops primarily by local Th1 host immune responses in the absence of parasite replication. AB - A single inoculation of mice with the live, attenuated Toxoplasma gondii uracil auxotroph strain cps1-1 induces long-lasting immunity against lethal challenge with hypervirulent strain RH. The mechanism for this robust immunity in the absence of parasite replication has not been addressed. The mechanism of long lasting immunity, the importance of route of immunization, cellular recruitment to the site of infection, and local and systemic inflammation were evaluated. Our results show that infection with cps1-1 elicits long-lasting CD8+ T cell- mediated immunity. We show that immunization with cps1-1-infected dendritic cells elicits long-lasting immunity. Intraperitoneal infection with cps1-1 induced a rapid influx of GR1+ neutrophils and two stages of GR1+CD68+ inflammatory monocyte infiltration into the site of inoculation. CD19+ B cells and CD3+ T cells steadily increase for 8 days after infection. CD8+ T cells were rapidly recruited to the site of infection and increased faster than CD4+ T cells. Surprisingly, cps1-1 infection induced high systemic levels of bioactive IL-12p70 and a very low level and transient systemic IFN-gamma. Furthermore, we show significant levels of these inflammatory cytokines were locally produced at the site of cps1-1 inoculation. These findings offer new insight into immunological mechanisms and local host responses to a non-replicating type I parasite infection associated with development of long-lasting immunity to Toxoplasma gondii. PMID- 19124751 TI - Fulminant lymphocytic choriomeningitis virus-induced inflammation of the CNS involves a cytokine-chemokine-cytokine-chemokine cascade. AB - Intracerebral inoculation of immunocompetent mice with lymphocytic choriomeningitis virus (LCMV) normally results in fatal CD8+ T cell mediated meningoencephalitis. However, in CXCL10-deficient mice, the virus-induced CD8+ T cell accumulation in the neural parenchyma is impaired, and only 30-50% of the mice succumb to the infection. Similar results are obtained in mice deficient in the matching chemokine receptor, CXCR3. Together, these findings point to a key role for CXCL10 in regulating the severity of the LCMV-induced inflammatory process. For this reason, we now address the mechanisms regulating the expression of CXCL10 in the CNS of LCMV-infected mice. Using mice deficient in type I IFN receptor, type II IFN receptor, or type II IFN, as well as bone marrow chimeras expressing CXCL10 only in resident cells or only in bone marrow-derived cells, we analyzed the up-stream regulation as well as the cellular source of CXCL10. We found that expression of CXCL10 initially depends on signaling through the type I IFN receptor, while late expression and up-regulation requires type II IFN produced by the recruited CD8+ T cells. Throughout the infection, the producers of CXCL10 are exclusively resident cells of the CNS, and astrocytes are the dominant expressors in the neural parenchyma, not microglial cells or recruited bone marrow-derived cell types. These results are consistent with a model suggesting a bidirectional interplay between resident cells of the CNS and the recruited virus-specific T cells with astrocytes as active participants in the local antiviral host response. PMID- 19124752 TI - Induction of proinflammatory cytokines in primary human macrophages by influenza A virus (H5N1) is selectively regulated by IFN regulatory factor 3 and p38 MAPK. AB - The hyperinduction of proinflammatory cytokines and chemokines such as TNF-alpha, IFN-beta, and CCL2/MCP-1 in primary human macrophages and respiratory epithelial cells by the highly pathogenic avian influenza H5N1 is believed to contribute to the unusual severity of human H5N1 disease. Here we show that TNF-alpha, IFN beta, and IFN-lambda1 are the key mediators directly induced by the H5N1 virus in primary human macrophages. In comparison with human influenza (H1N1), the H5N1 virus more strongly activated IFN regulatory factor 3 (IRF3). IRF3 knockdown and p38 kinase inhibition separately and in combination led to a substantial reduction of IFN-beta, IFN-lambda1, and MCP-1 but only to a partial reduction of TNF-alpha. IRF3 translocation was independent of p38 kinase activity, indicating that IRF3 and p38 kinase are distinct pathways leading to cytokine production by H5N1 virus. We conclude that IRF3 and p38 kinase separately and predominantly contribute to H5N1-mediated induction of IFN-beta, IFN-lambda1, and MCP-1 but only partly control TNF-alpha induction. A more precise identification of the differences in the regulation of TNF-alpha and IFN-beta could provide novel targets for the design of therapeutic strategies for severe human H5N1 influenza and also for treating other causes of acute respiratory distress syndrome. PMID- 19124753 TI - Type I IFN-mediated protection of macrophages and dendritic cells secures control of murine coronavirus infection. AB - The swift production of type I IFNs is one of the fundamental aspects of innate immune responses against viruses. Plasmacytoid dendritic cell-derived type I IFNs are of prime importance for the initial control of highly cytopathic viruses such as the mouse hepatitis virus (MHV). The aim of this study was to determine the major target cell populations of this first wave of type I IFNs. Generation of bone marrow-chimeric mice expressing the type I IFN receptor (IFNAR) on either hemopoietic or non-bone marrow-derived cells revealed that the early control of MHV depended mainly on IFNAR expression on hemopoietic cells. To establish which cell population responds most efficiently to type I IFNs, mice conditionally deficient for the IFNAR on different leukocyte subsets were infected with MHV. This genetic analysis revealed that IFNAR expression on LysM+ macrophages and CD11c+ dendritic cells was most important for the early containment of MHV within secondary lymphoid organs and to prevent lethal liver disease. This study identifies type I IFN-mediated cross-talk between plasmacytoid dendritic cells on one side and macrophages and conventional dendritic cells on the other, as an essential cellular pathway for the control of fatal cytopathic virus infection. PMID- 19124754 TI - IL-10 dampens TNF/inducible nitric oxide synthase-producing dendritic cell mediated pathogenicity during parasitic infection. AB - Antiparasite responses are associated with the recruitment of monocytes that differentiate to macrophages and dendritic cells at the site of infection. Although classically activated monocytic cells are assumed to be the major source of TNF and NO during Trypanosoma brucei brucei infection, their cellular origin remains unclear. In this study, we show that bone marrow-derived monocytes accumulate and differentiate to TNF/inducible NO synthase-producing dendritic cells (TIP-DCs) in the spleen, liver, and lymph nodes of T. brucei brucei infected mice. Although TIP-DCs have been shown to play a beneficial role in the elimination of several intracellular pathogens, we report that TIP-DCs, as a major source of TNF and NO in inflamed organs, could contribute actively to tissue damage during the chronic stage of T. brucei brucei infection. In addition, the absence of IL-10 leads to enhanced differentiation of monocytes to TIP-DCs, resulting in exacerbated pathogenicity and early death of the host. Finally, we demonstrate that sustained production of IL-10 following IL-10 gene delivery treatment with an adeno-associated viral vector to chronically infected mice limits the differentiation of monocytes to TIP-DCs and protects the host from tissue damage. PMID- 19124755 TI - Dectin-2 recognition of house dust mite triggers cysteinyl leukotriene generation by dendritic cells. AB - House dust mites are a significant source of airborne allergen worldwide, but there is little understanding of how they so potently generate allergic inflammation. We found that extracts from the house dust mites Dermatophagoides farinae (Df) and Dermatophagoides pteronyssinus and from the mold Aspergillus fumigatus stimulated a rapid and robust production of cysteinyl leukotrienes (cys LTs), proinflammatory lipid mediators, from mouse bone marrow-derived dendritic cells (BMDCs). Con A affinity chromatography of the Df extract revealed that the relevant ligand is a glycan(s), suggesting stimulation via a dendritic cell (DC) lectin receptor. Cys-LT production in BMDCs from wild-type mice was inhibited by spleen tyrosine kinase (Syk) inhibitors and was abolished in BMDCs from FcRgamma /- mice, implicating either Dectin-2 or DC immunoactivating receptor. Transfection of each receptor in bone marrow-derived mast cells revealed that only Dectin-2 mediates cys-LT production by Df, Dermatophagoides pteronyssinus, and Aspergillus fumigatus. Lentiviral knockdown of Dectin-2 in BMDCs attenuated Df extract-elicited cys-LT generation, thereby identifying Dectin-2 as the receptor. Lung CD11c+ cells, but not peritoneal or alveolar macrophages, also generated cys-LTs in response to Df. These findings place Dectin-2 among the C type lectin receptors that activate arachidonic acid metabolism and identify the Dectin-2/FcRgamma/Syk/cys-LT axis as a novel mechanism by which three potent indoor allergens may activate innate immune cells to promote allergic inflammation. PMID- 19124756 TI - P2Y6 receptors require an intact cysteinyl leukotriene synthetic and signaling system to induce survival and activation of mast cells. AB - Cysteinyl leukotrienes (cys-LTs) induce inflammatory responses through type 1 (CysLT1R) and type 2 (CysLT2R) cys-LT receptors and activate mast cells in vitro. We previously demonstrated that cys-LTs cross-desensitized IL-4-primed primary human mast cells (hMCs) to stimulation with the nucleotide uridine diphosphate (UDP). We now report that hMCs, mouse bone marrow-derived mast cells (mBMMCs), and the human MC line LAD2 all express UDP-selective P2Y6 receptors that cooperate with CysLT1R to promote cell survival and chemokine generation by a pathway involving reciprocal ligand-mediated cross-talk. Leukotriene (LT) D4, the most potent CysLT1R ligand, and UDP both induced phosphorylation of ERK and prolonged the survival of cytokine-starved hMCs and mBMMCs. ERK activation and cytoprotection in response to either ligand were attenuated by treatment of the cells with a selective P2Y6 receptor antagonist (MRS2578), which did not interfere with signaling through recombinant CysLT1R. Surprisingly, both UDP and LTD4-mediated ERK activation and cytoprotection were absent in mBMMCs lacking CysLT1R and the biosynthetic enzyme LTC4 synthase, implying a requirement for a cys-LT-mediated autocrine loop. In IL-4-primed LAD2 cells, LTD4 induced the generation of MIP-1beta, a response blocked by short hairpin RNA-mediated knockdown of CysLT1R or P2Y6 receptors, but not of CysLT2R. Thus, CysLT1R and P2Y6 receptors, which are coexpressed on many cell types of innate immunity, reciprocally amplify one another's function in mast cells through endogenous ligands. PMID- 19124757 TI - Increased reactivity of dendritic cells from aged subjects to self-antigen, the human DNA. AB - Diminished immune functions and chronic inflammation are hallmarks of aging. The underlying causes are not well understood. In this investigation, we show an increased reactivity of dendritic cells (DCs) from aged subjects to self-Ags as one of the potential mechanisms contributing to age-associated inflammation. Consistent with this, DCs from aged subjects display increased reactivity to intracellular human DNA, a self-Ag, by secreting enhanced quantities of type I IFN and IL-6 compared with the DCs from young subjects. Furthermore, this is accompanied by an increased up-regulation of costimulatory molecules CD80 and CD86. These DNA-primed DCs from aged subjects enhanced T cell proliferation compared with the young subjects, further substantiating our findings. Investigations of signaling mechanisms revealed that DNA-stimulated DCs from aged subjects displayed a significantly higher level of IFN regulatory factor-3 and NF kappaB activity compared with their young counterparts. More importantly, DCs from aged subjects displayed a higher level of NF-kappaB activation at the basal level, suggesting an increased state of activation. This activated state of DCs may be responsible for their increased reactivity to self-Ags such as DNA, which in turn contributes to the age-associated chronic inflammation. PMID- 19124758 TI - Differential use of CARD9 by dectin-1 in macrophages and dendritic cells. AB - The pattern recognition receptors TLR2 and Dectin-1 play key roles in coordinating the responses of macrophages and dendritic cells (DC) to fungi. Induction of proinflammatory cytokines is instructed by signals from both TLR2 and Dectin-1. A recent report identified a role for CARD9 in innate anti-fungal responses, demonstrating CARD9-Bcl10-mediated activation of NF-kappaB and proinflammatory cytokine induction in murine bone marrow-derived DC stimulated via Dectin-1. We now report that Dectin-1-CARD9 signals fail to activate NF kappaB and drive TNF-alpha induction in murine bone marrow-derived macrophages. However, priming of bone marrow-derived macrophages with GM-CSF or IFN-gamma permits Dectin-1-CARD9-mediated TNF-alpha induction. Analysis of other macrophage/DC populations revealed further variation in the ability of Dectin-1 CARD9 signaling to drive TNF-alpha production. Resident peritoneal cells and alveolar macrophages produce TNF-alpha upon Dectin-1 ligation, while thioglycollate-elicited peritoneal macrophages and Flt3L-derived DC do not. We present data demonstrating that CARD9 is recruited to phagosomes via its CARD domain where it enhances TLR-induced cytokine production even in cells in which Dectin-1 is insufficient to drive cytokine production. In such cells, Dectin-1, CARD9, and Bcl10 levels are not limiting, and data indicate that these cells express additional factors that restrict Dectin-1-CARD9 signaling for TNF-alpha induction. PMID- 19124760 TI - Galectin-3 deficiency reduces the severity of experimental autoimmune encephalomyelitis. AB - Galectin-3 (Gal-3) is a member of the beta-galactoside-binding lectin family and plays an important role in inflammation. However, the precise role of Gal-3 in autoimmune diseases remains obscure. We have investigated the functional role of Gal-3 in experimental autoimmune encephalomyelitis (EAE) following immunization with myelin oligodendrocyte glycoprotein (MOG)35-55 peptide. Gal-3 deficient (Gal 3-/-) mice developed significantly milder EAE and markedly reduced leukocyte infiltration in the CNS compared with similarly treated wild-type (WT) mice. Gal 3-/- mice also contained fewer monocytes and macrophages but more apoptotic cells in the CNS than did WT mice. Following Ag stimulation in vitro, lymph node cells from the immunized Gal-3-/- mice produced less IL-17 and IFN-gamma than did those of the WT mice. In contrast, Gal-3-/- mice produced more serum IL-10, IL-5, and IL-13 and contained higher frequency of Foxp3+ regulatory T cells in the CNS than did the WT mice. Furthermore, bone marrow-derived dendritic cells from Gal-3-/- mice produced more IL-10 in response to LPS or bacterial lipoprotein than did WT marrow-derived dendritic cells. Moreover, Gal-3-/- dendritic cells induced Ag specific T cells to produce more IL-10, IL-5, and IL-12, but less IL-17, than did WT dendritic cells. Taken together, our data demonstrate that Gal-3 plays an important disease-exacerbating role in EAE through its multifunctional roles in preventing cell apoptosis and increasing IL-17 and IFN-gamma synthesis, but decreasing IL-10 production. PMID- 19124759 TI - Mobilization and margination of bone marrow Gr-1high monocytes during subclinical endotoxemia predisposes the lungs toward acute injury. AB - The specialized role of mouse Gr-1(high) monocytes in local inflammatory reactions has been well documented, but the trafficking and responsiveness of this subset during systemic inflammation and their contribution to sepsis-related organ injury has not been investigated. Using flow cytometry, we studied monocyte subset margination to the pulmonary microcirculation during subclinical endotoxemia in mice and investigated whether marginated monocytes contribute to lung injury in response to further septic stimuli. Subclinical low-dose i.v. LPS induced a rapid (within 2 h), large-scale mobilization of bone marrow Gr-1high monocytes and their prolonged margination to the lungs. With secondary LPS challenge, membrane TNF expression on these premarginated monocytes substantially increased, indicating their functional priming in vivo. Zymosan challenge produced small increases in pulmonary vascular permeability, which were markedly enhanced by the preadministration of low-dose LPS. The LPS-zymosan-induced permeability increases were effectively abrogated by pretreatment (30 min before zymosan challenge) with the platelet-activating factor antagonist WEB 2086 in combination with the phosphatidylcholine-phospholipase C inhibitor D609, suggesting the involvement of platelet-activating factor/ceramide-mediated pathways in this model. Depletion of monocytes (at 18 h after clodronate-liposome treatment) significantly attenuated the LPS-zymosan-induced permeability increase. However, restoration of normal LPS-induced Gr-1high monocyte margination to the lungs (at 48 h after clodronate-liposome treatment) resulted in the loss of this protective effect. These results demonstrate that mobilization and margination of Gr-1high monocytes during subclinical endotoxemia primes the lungs toward further septic stimuli and suggest a central role for this monocyte subset in the development of sepsis-related acute lung injury. PMID- 19124761 TI - Pulmonary V gamma 4+ gamma delta T cells have proinflammatory and antiviral effects in viral lung disease. AB - Host defenses, while effecting viral clearance, contribute substantially to inflammation and disease. This double action is a substantial obstacle to the development of safe and effective vaccines against many agents, particularly respiratory syncytial virus (RSV). RSV is a common cold virus and the major cause of infantile bronchiolitis worldwide. The role of alphabeta T cells in RSV-driven immunopathology is well studied, but little is known about the role of "unconventional" T cells. During primary RSV challenge of BALB/c mice, some Vgamma7+ gammadelta T cells were present; however, immunization with a live vaccinia vector expressing RSV F protein substantially enhanced Vgamma4+ gammadelta T cell influx after RSV infection. Harvested early, these cells produced IFN-gamma, TNF, and RANTES after ex vivo stimulation. By contrast, those recruited 5 days after challenge made IL-4, IL-5, and IL-10. Depletion of gammadelta T cells in vivo reduced lung inflammation and disease severity and slightly increased peak viral replication but did not prevent viral clearance. These studies demonstrate a novel role for gammadelta T cells in the development of immunopathology and cellular influx into the lungs after immunization and RSV challenge. Though a minor population, gammadelta T cells have a critical influence on disease and are an attractive interventional target in the alleviation of viral lung disease. PMID- 19124762 TI - Acquisition of anergy to proinflammatory cytokines in nonimmune cells through endoplasmic reticulum stress response: a mechanism for subsidence of inflammation. AB - Acute endoplasmic reticulum (ER) stress causes induction of inflammatory molecules via activation of NF-kappaB. However, we found that, under ER stress conditions, renal mesangial cells acquire anergy to proinflammatory stimuli. Priming of the cells with ER stress inducers (tunicamycin, thapsigargin, A23187, and AB5 subtilase cytotoxin) caused blunted induction of MCP-1 in response to TNF alpha, IL-1beta, macrophage-derived factors, or bystander macrophages. The magnitude of suppression was closely correlated with the level of GRP78, an endogenous indicator of ER stress. The suppression of MCP-1 under ER stress conditions was reversible and observed in general regardless of cell types or triggers of ER stress. The decrease in the level of MCP-1 mRNA was ascribed to transcriptional suppression via unexpected inhibition of NF-kappaB, but not to accelerated mRNA degradation. Subsequent experiments revealed that TNFR associated factor 2, an essential component for TNF-alpha signaling, was down regulated by ER stress. We also found that, under ER stress conditions, expression of NF-kappaB suppressor A20 was induced. Overexpression of A20 resulted in suppression of cytokine-triggered NF-kappaB activation and knockdown of A20 by RNA interference significantly attenuated induction of anergy by ER stress. In contrast, other ER stress-inducible/-related molecules that may suppress NF-kappaB (e.g., GRP78, NO, reactive oxygen species, and IkappaB) were not involved in the inhibitory effects of ER stress. These results elucidated ER stress-dependent mechanisms by which nonimmune cells acquire anergy to inflammatory stimuli under pathological situations. This self-defense machinery may play a role in halting progression of acute inflammation and in its spontaneous subsidence. PMID- 19124764 TI - Rapid degranulation of NK cells following activation by HIV-specific antibodies. AB - Ab-dependent cellular cytotoxicity (ADCC) Abs stimulate NK cell effector functions and play a role in protecting from and controlling viral infections. We characterized ADCC Abs in a cross-sectional cohort of 80 HIV-infected subjects not on antiretroviral therapy. We analyzed ADCC response by killing fluorescently labeled target cells, as well as expression of IFN-gamma and the degranulation marker CD107a from activated NK cells as measured by a novel intracellular cytokine assay. HIV-specific ADCC directed toward Envelope proteins were present in the majority of 80 untreated HIV-infected individuals measured by killing function. Similarly, most subjects had HIV-specific Abs that mediated degranulation or cytokine expression by NK cells. Interestingly, there was a poor correlation between ADCC-mediated killing of fluorescently labeled whole Envelope protein-pulsed cell lines and Ab-mediated expression of IFN-gamma by NK cells. However, in contrast to healthy donor NK cells, autologous patient NK cells more effectively degranulated granzyme B in response to ADCC activation. Activation of NK cells in response to stimulation by HIV-specific Abs occurs at least as rapidly as activation of Gag-specific CTLs. Our studies highlight the complexity of ab-mediated NK cell activation in HIV infection, and suggest new avenues toward studying the utility of ADCC in controlling HIV infection. PMID- 19124763 TI - Potent induction of IFN-alpha and chemokines by autoantibodies in the cerebrospinal fluid of patients with neuropsychiatric lupus. AB - Neuropsychiatric disease in systemic lupus erythematosus (NPSLE) is a poorly understood, but potentially fatal, disease manifestation. A pathogenetic role for autoantibodies is suspected, but the mechanism is unclear. Since immune complexes in SLE can stimulate IFN-alpha and there is strong evidence in humans and in mice that IFN-alpha can cause neuropsychiatric manifestations, we asked whether NPSLE patient serum and/or cerebrospinal fluid (CSF) contain abnormally high IFN-alpha inducing activity. In a bioassay containing plasmacytoid dendritic cells and a source of Ag, NPSLE CSF induced significantly higher IFN-alpha compared with CSF from patients with multiple sclerosis or other autoimmune disease controls. When normalized for IgG concentration, NPSLE CSF was 800-fold more potent at inducing IFN-alpha compared with paired serum due to inhibitors present in serum. Analysis of Ig-deficient patient serum, depletion of IgG from normal serum, as well as addition of purified IgG to NPSLE CSF and serum in the bioassays revealed that one inhibitor was contained within the IgG fraction itself. In addition to IFN alpha, immune complexes formed by CSF autoantibodies produced significantly increased levels of IFN-gamma-inducible protein 10 (IP-10/CXCL), IL-8, and MCP-1, all of which have been reported to be elevated in CSF from NPSLE patients. Taken together, these findings are consistent with a two-step model of NPSLE whereby CSF autoantibodies bind to Ags released by neurocytotoxic Abs or other brain cell injury, and the resulting immune complexes stimulate IFN-alpha and proinflammatory cytokines and chemokines. PMID- 19124766 TI - Gamma flicker triggers attentional selection without awareness. AB - Gamma band modulations in neural activity have been proposed to mediate attentional processes. To support a causal link between gamma activity and attentional selection, we attempt to evoke gamma oscillations by a 50-Hz subliminal flicker. We find that a subliminal 50-Hz flicker at a target location, before target presentation, speeds up and enhances target detection and discrimination. This effect is specific to the middle of the gamma range because it is not evident at <35-Hz flicker. It requires 300 ms to build up, dissipates within 250 ms of flicker offset, and shows a tendency to invert after 500 ms. The results are discussed in relation to a role for gamma band neural synchrony in the allocation of visual attention. PMID- 19124765 TI - Mucosally delivered Salmonella typhi expressing the Yersinia pestis F1 antigen elicits mucosal and systemic immunity early in life and primes the neonatal immune system for a vigorous anamnestic response to parenteral F1 boost. AB - Neonates respond poorly to conventional vaccines. This has been attributed, in part, to the immaturity of neonatal dendritic cells that lack full capacity for Ag presentation and T cell stimulation. We engineered an attenuated Salmonella Typhi strain to express and export the F1 Ag of Y. pestis (S. Typhi(F1)) and investigated its immunogenicity early in life using a heterologous prime-boost regimen. Newborn mice primed intranasally with a single dose of S. Typhi(F1) elicited mucosal Ab- and IFN-gamma-secreting cells 1 wk after immunization. They also developed a potent and fast anamnestic response to a subsequent parenteral boost with F1-alum, which surpassed those of mice primed and boosted with S. Typhi(F1) or F1-alum. Neonatal priming with S. Typhi(F1), as opposed to priming with F1-alum, resulted in a more balanced IgG2a/IgG1 profile, enhanced avidity maturation and stimulation of B memory cells, and strong Th1-type cell-mediated immunity. S. Typhi(F1) enhanced the activation and maturation of neonatal CD11c+ dendritic cells, shown by increased expression of CD80, CD86, CD40, and MHC-II cell surface markers and production of proinflammatory cytokines IL-12, TNF alpha, IL-6, and MCP-1. S. Typhi(F1)-stimulated neonatal DC had improved capacity for Ag presentation and T cell stimulation in vitro and induced F1-specific CD4+ and CD8+ T cell responses when adoptively transferred to newborn mice. Mucosal immunization with S. Typhi expressing a foreign Ag effectively primes the neonatal immune system for potent, fast, and broader responses to a parenteral Ag boost. Such a strategy can prevent infectious diseases, including those considered biowarfare threats, early in life. PMID- 19124767 TI - Force amplification response of actin filaments under confined compression. AB - Actin protein is a major component of the cell cytoskeleton, and its ability to respond to external forces and generate propulsive forces through the polymerization of filaments is central to many cellular processes. The mechanisms governing actin's abilities are still not fully understood because of the difficulty in observing these processes at a molecular level. Here, we describe a technique for studying actin-surface interactions by using a surface forces apparatus that is able to directly visualize and quantify the collective forces generated when layers of noninterconnected, end-tethered actin filaments are confined between 2 (mica) surfaces. We also identify a force-response mechanism in which filaments not only stiffen under compression, which increases the bending modulus, but more importantly generates opposing forces that are larger than the compressive force. This elastic stiffening mechanism appears to require the presence of confining surfaces, enabling actin filaments to both sense and respond to compressive forces without additional mediating proteins, providing insight into the potential role compressive forces play in many actin and other motor protein-based phenomena. PMID- 19124768 TI - Tyrosine phosphorylation of the BRI1 receptor kinase emerges as a component of brassinosteroid signaling in Arabidopsis. AB - Brassinosteroids (BRs) are essential growth-promoting hormones that regulate many aspects of plant growth and development. Two leucine-rich repeat receptor-like kinases (LRR-RLKs) are involved in BR perception and signal transduction: brassinosteroid insensitive 1 (BRI1), which is the BR receptor, and its coreceptor BRI1-associated kinase 1 (BAK1). Both proteins are classified as serine/threonine protein kinases, but here we report that recombinant cytoplasmic domains of BRI1 and BAK1 also autophosphorylate on tyrosine residues and thus are dual-specificity kinases. With BRI1, Tyr-831 and Tyr-956 are identified as autophosphorylation sites in vitro and in vivo. Interestingly, Tyr-956 in kinase subdomain V is essential for activity, because the Y956F mutant is catalytically inactive and thus this site cannot be simply manipulated by mutagenesis. In contrast, Tyr-831 in the juxtamembrane domain is not essential for kinase activity but plays an important role in BR signaling in vivo, because expression of BRI1(Y831F)-Flag in transgenic bri1-5 plants results in plants with larger leaves (but altered leaf shape) and early flowering relative to plants expressing wild-type BRI1-Flag. Acidic substitutions of Tyr-831 restored normal leaf size (but not shape) and normal flowering time. This is an example where a specific tyrosine residue has been shown to play an important role in vivo in plant receptor kinase function. Interestingly, 6 additional LRR-RLKs (of the 23 tested) were also found to autophosphorylate on tyrosine in addition to serine and threonine, suggesting that tyrosine signaling should be considered with other plant receptor kinases as well. PMID- 19124769 TI - A DNA damage response in Escherichia coli involving the alternative sigma factor, RpoS. AB - We isolated an Escherichia coli mutant in the iraD gene, sensitive to various forms of DNA damage. Our data are consistent with the function of IraD to promote accumulation of the alternative transcription sigma factor, RpoS, by binding to the adaptor RssB protein that targets RpoS for degradation. Our results demonstrate the physiological importance of this mode of regulation for DNA damage tolerance. Although RpoS is best known for its regulation of genes induced in stationary phase, our work underscores the importance of the RpoS regulon in a DNA damage response in actively growing cells. We show that iraD transcription is induced by DNA damage by a mechanism independent of the SOS response. The IraD and SOS regulatory pathways appear to act synergistically to ensure survival of cells faced with oxidative or DNA damaging stress during cellular growth. PMID- 19124770 TI - Phytohormone-based activity mapping of insect herbivore-produced elicitors. AB - In response to insect attack, many plants exhibit dynamic biochemical changes, resulting in the induced production of direct and indirect defenses. Elicitors present in herbivore oral secretions are believed to positively regulate many inducible plant defenses; however, little is known about the specificity of elicitor recognition in plants. To investigate the phylogenic distribution of elicitor activity, we tested representatives from three different elicitor classes on the time course of defense-related phytohormone production, including ethylene (E), jasmonic acid (JA), and salicylic acid, in a range of plant species spanning angiosperm diversity. All families examined responded to at least one elicitor class with significant increases in E and JA production within 1 to 2 h after treatment, yet elicitation activity among species was highly idiosyncratic. The fatty-acid amino acid conjugate volicitin exhibited the widest range of phytohormone and volatile inducing activity, which spanned maize (Zea mays), soybean (Glycine max), and eggplant (Solanum melongena). In contrast, the activity of inceptin-related peptides, originally described in cowpea (Vigna unguiculata), was limited even within the Fabaceae. Similarly, caeliferin A16:0, a disulfooxy fatty acid from grasshoppers, was the only elicitor with demonstrable activity in Arabidopsis thaliana. Although precise mechanisms remain unknown, the unpredictable nature of elicitor activity between plant species supports the existence of specific receptor-ligand interactions mediating recognition. Despite the lack of an ideal plant model for studying the action of numerous elicitors, E and JA exist as highly conserved and readily quantifiable markers for future discoveries in this field. PMID- 19124771 TI - Exploration dynamics in evolutionary games. AB - Evolutionary game theory describes systems where individual success is based on the interaction with others. We consider a system in which players unconditionally imitate more successful strategies but sometimes also explore the available strategies at random. Most research has focused on how strategies spread via genetic reproduction or cultural imitation, but random exploration of the available set of strategies has received less attention so far. In genetic settings, the latter corresponds to mutations in the DNA, whereas in cultural evolution, it describes individuals experimenting with new behaviors. Genetic mutations typically occur with very small probabilities, but random exploration of available strategies in behavioral experiments is common. We term this phenomenon "exploration dynamics" to contrast it with the traditional focus on imitation. As an illustrative example of the emerging evolutionary dynamics, we consider a public goods game with cooperators and defectors and add punishers and the option to abstain from the enterprise in further scenarios. For small mutation rates, cooperation (and punishment) is possible only if interactions are voluntary, whereas moderate mutation rates can lead to high levels of cooperation even in compulsory public goods games. This phenomenon is investigated through numerical simulations and analytical approximations. PMID- 19124772 TI - O6-methylguanine-induced cell death involves exonuclease 1 as well as DNA mismatch recognition in vivo. AB - Alkylation-induced O(6)-methylguanine (O(6)MeG) DNA lesions can be mutagenic or cytotoxic if unrepaired by the O(6)MeG-DNA methyltransferase (Mgmt) protein. O(6)MeG pairs with T during DNA replication, and if the O(6)MeG:T mismatch persists, a G:C to A:T transition mutation is fixed at the next replication cycle. O(6)MeG:T mismatch detection by MutSalpha and MutLalpha leads to apoptotic cell death, but the mechanism by which this occurs has been elusive. To explore how mismatch repair mediates O(6)MeG-dependent apoptosis, we used an Mgmt-null mouse model combined with either the Msh6-null mutant (defective in mismatch recognition) or the Exo1-null mutant (impaired in the excision step of mismatch repair). Mouse embryonic fibroblasts and bone marrow cells derived from Mgmt-null mice were much more alkylation-sensitive than wild type, as expected. However, ablation of either Msh6 or Exo1 function rendered these Mgmt-null cells just as resistant to alkylation-induced cytotoxicity as wild-type cells. Rapidly proliferating tissues in Mgmt-null mice (bone marrow, thymus, and spleen) are extremely sensitive to apoptosis induced by O(6)MeG-producing agents. Here, we show that ablation of either Msh6 or Exo1 function in the Mgmt-null mouse renders these rapidly proliferating tissues alkylation-resistant. However, whereas the Msh6 defect confers total alkylation resistance, the Exo1 defect leads to a variable tissue-specific alkylation resistance phenotype. Our results indicate that Exo1 plays an important role in the induction of apoptosis by unrepaired O(6)MeGs. PMID- 19124774 TI - Turning back from the brink: detecting an impending regime shift in time to avert it. AB - Ecological regime shifts are large, abrupt, long-lasting changes in ecosystems that often have considerable impacts on human economies and societies. Avoiding unintentional regime shifts is widely regarded as desirable, but prediction of ecological regime shifts is notoriously difficult. Recent research indicates that changes in ecological time series (e.g., increased variability and autocorrelation) could potentially serve as early warning indicators of impending shifts. A critical question, however, is whether such indicators provide sufficient warning to adapt management to avert regime shifts. We examine this question using a fisheries model, with regime shifts driven by angling (amenable to rapid reduction) or shoreline development (only gradual restoration is possible). The model represents key features of a broad class of ecological regime shifts. We find that if drivers can only be manipulated gradually management action is needed substantially before a regime shift to avert it; if drivers can be rapidly altered aversive action may be delayed until a shift is underway. Large increases in the indicators only occur once a regime shift is initiated, often too late for management to avert a shift. To improve usefulness in averting regime shifts, we suggest that research focus on defining critical indicator levels rather than detecting change in the indicators. Ideally, critical indicator levels should be related to switches in ecosystem attractors; we present a new spectral density ratio indicator to this end. Averting ecological regime shifts is also dependent on developing policy processes that enable society to respond more rapidly to information about impending regime shifts. PMID- 19124773 TI - An overlooked pink species of land iguana in the Galapagos. AB - Despite the attention given to them, the Galapagos have not yet finished offering evolutionary novelties. When Darwin visited the Galapagos, he observed both marine (Amblyrhynchus) and land (Conolophus) iguanas but did not encounter a rare pink black-striped land iguana (herein referred to as "rosada," meaning "pink" in Spanish), which, surprisingly, remained unseen until 1986. Here, we show that substantial genetic isolation exists between the rosada and syntopic yellow forms and that the rosada is basal to extant taxonomically recognized Galapagos land iguanas. The rosada, whose present distribution is a conundrum, is a relict lineage whose origin dates back to a period when at least some of the present-day islands had not yet formed. So far, this species is the only evidence of ancient diversification along the Galapagos land iguana lineage and documents one of the oldest events of divergence ever recorded in the Galapagos. Conservation efforts are needed to prevent this form, identified by us as a good species, from extinction. PMID- 19124775 TI - Forecasting United States mortality using cohort smoking histories. AB - In this paper, we introduce a recently established relationship between cohort smoking patterns and adult mortality into mortality projections for the United States. In particular, we incorporate a variable representing the intensity of smoking within a cohort into the original Lee-Carter projection model. The introduction of this variable accounts for important anomalies in the recent age/sex pattern of mortality change and enables the use of a common temporal trend of mortality change for the 2 sexes. We project age-specific mortality rates for men and women at ages 50-84 between 2004 and 2034 in the United States. Because of reductions in smoking that have already occurred or can be reliably projected, mortality is projected to decline much faster when smoking is introduced into the model. PMID- 19124776 TI - The phage abortive infection system, ToxIN, functions as a protein-RNA toxin antitoxin pair. AB - Various mechanisms exist that enable bacteria to resist bacteriophage infection. Resistance strategies include the abortive infection (Abi) systems, which promote cell death and limit phage replication within a bacterial population. A highly effective 2-gene Abi system from the phytopathogen Erwinia carotovora subspecies atroseptica, designated ToxIN, is described. The ToxIN Abi system also functions as a toxin-antitoxin (TA) pair, with ToxN inhibiting bacterial growth and the tandemly repeated ToxI RNA antitoxin counteracting the toxicity. TA modules are currently divided into 2 classes, protein and RNA antisense. We provide evidence that ToxIN defines an entirely new TA class that functions via a novel protein RNA mechanism, with analogous systems present in diverse bacteria. Despite the debated role of TA systems, we demonstrate that ToxIN provides viral resistance in a range of bacterial genera against multiple phages. This is the first demonstration of a novel mechanistic class of TA systems and of an Abi system functioning in different bacterial genera, both with implications for the dynamics of phage-bacterial interactions. PMID- 19124777 TI - Cryo-electron tomography of mouse hepatitis virus: Insights into the structure of the coronavirion. AB - Coronaviruses are enveloped viruses containing the largest reported RNA genomes. As a result of their pleomorphic nature, our structural insight into the coronavirion is still rudimentary, and it is based mainly on 2D electron microscopy. Here we report the 3D virion structure of coronaviruses obtained by cryo-electron tomography. Our study focused primarily on the coronavirus prototype murine hepatitis virus (MHV). MHV particles have a distinctly spherical shape and a relatively homogenous size ( approximately 85 nm envelope diameter). The viral envelope exhibits an unusual thickness (7.8 +/- 0.7 nm), almost twice that of a typical biological membrane. Focal pairs revealed the existence of an extra internal layer, most likely formed by the C-terminal domains of the major envelope protein M. In the interior of the particles, coiled structures and tubular shapes are observed, consistent with a helical nucleocapsid model. Our reconstructions provide no evidence of a shelled core. Instead, the ribonucleoprotein seems to be extensively folded onto itself, assuming a compact structure that tends to closely follow the envelope at a distance of approximately 4 nm. Focal contact points and thread-like densities connecting the envelope and the ribonucleoprotein are revealed in the tomograms. Transmissible gastroenteritis coronavirion tomograms confirm all the general features and global architecture observed for MHV. We propose a general model for the structure of the coronavirion in which our own and published observations are combined. PMID- 19124780 TI - Medical errors, apologies and apology laws. PMID- 19124778 TI - Scaling and self-organized criticality in proteins II. AB - The complexity of proteins is substantially simplified by regarding them as archetypical examples of self-organized criticality (SOC). To test this idea and to elaborate it, this article applies the Moret-Zebende (MZ) SOC hydrophobicity scale to transport repeat proteins of the HEAT superfamily, importin beta, and transportin, as well as the export protein Cse1p, and their ubiquitous cargo manager Ran. The difference between the MZ scale and conventional hydrophobicity scales reflects long-range conformational forces that are central to protein functionality. These compete with long-range Coulomb forces associated with cationic and anionic side chains in a revealing way. PMID- 19124781 TI - The controversy over the efficacy of pneumococcal vaccine. PMID- 19124782 TI - Clinical trials: the muddled Canadian landscape. PMID- 19124783 TI - Legumes, lemons and streptomycin: a short history of the clinical trial. PMID- 19124784 TI - Physician allocation system in Quebec inhibits recruitment. PMID- 19124785 TI - Ex-provincial health minister sees red. PMID- 19124788 TI - Arctic health research. PMID- 19124786 TI - Somali symbiosis, part 3: how it affects me. PMID- 19124789 TI - Comparison of prognosis for men with type 2 diabetes mellitus and men with cardiovascular disease. AB - BACKGROUND: People with type 2 diabetes mellitus are at high risk for cardiovascular disease. In some studies, the mortality rate among people with this condition has been equivalent to that among people with cardiovascular disease. We compared cardiovascular mortality between incident cases of diabetes and cardiovascular disease. METHODS: The study population was part of a random sample of 4376 men from Quebec, Canada, aged 35 to 64 years, who did not have cardiovascular disease in 1974 and who were followed until 1998. Three groups of incident cases were identified: diabetes without cardiovascular disease, first cardiovascular event (myocardial infarction, unstable angina or stroke) without diabetes, and both cardiovascular disease and diabetes. These cases were age matched to a control group without diabetes or cardiovascular disease. RESULTS: During the 24-year follow-up period, new diabetes without cardiovascular disease was documented in 137 men. A first cardiovascular event without diabetes was documented in 527 men. Relative to the 627 controls, men with 1 of the 2 diseases of interest had higher cardiovascular mortality (age-adjusted relative risk [RR] 3.11, 95% confidence interval [CI] 1.96-4.92) for those with diabetes and 4.46 (95% CI 3.15-6.30) for those with cardiovascular disease). However, within the first 5 years after diagnosis, men with cardiovascular disease had higher cardiovascular mortality than men with diabetes (age-adjusted RR 2.03, 95% CI 1.01-4.08). INTERPRETATION: Men with isolated type 2 diabetes and men with isolated cardiovascular disease had similar cardiovascular mortality rates several years after initial diagnosis of either condition. These findings reinforce the need to prevent and optimally manage diabetes and cardiovascular disease. PMID- 19124791 TI - Acute monoarthritis: what is the cause of my patient's painful swollen joint? PMID- 19124790 TI - Efficacy of pneumococcal vaccination in adults: a meta-analysis. AB - BACKGROUND: Clinical trials and meta-analyses have produced conflicting results of the efficacy of unconjugated pneumococcal polysaccharide vaccine in adults. We sought to evaluate the vaccine's efficacy on clinical outcomes as well as the methodologic quality of the trials. METHODS: We searched several databases and all bibliographies of reviews and meta-analyses for clinical trials that compared pneumococcal polysaccharide vaccine with a control. We examined rates of pneumonia and death, taking the methodologic quality of the trials into consideration. RESULTS: We included 22 trials involving 101 507 participants: 11 trials reported on presumptive pneumococcal pneumonia, 19 on all-cause pneumonia and 12 on all-cause mortality. The current 23-valent vaccine was used in 8 trials. The relative risk (RR) was 0.64 (95% confidence interval [CI] 0.43-0.96) for presumptive pneumococcal pneumonia and 0.73 (95% CI 0.56-0.94) for all-cause pneumonia. There was significant heterogeneity between the trials reporting on presumptive pneumonia (I(2) = 74%, p < 0.001) and between those reporting on all cause pneumonia (I(2) = 90%, p < 0.001). The RR for all-cause mortality was 0.97 (95% CI 0.87-1.09), with moderate heterogeneity between trials (I(2) = 44%, p = 0.053). Trial quality, especially regarding double blinding, explained a substantial proportion of the heterogeneity in the trials reporting on presumptive pneumonia and all-cause pneumonia. There was little evidence of vaccine protection in trials of higher methodologic quality (RR 1.20, 95% CI 0.75 1.92, for presumptive pneumonia; and 1.19, 95% CI 0.95-1.49, for all-cause pneumonia in double-blind trials; p for heterogeneity > 0.05). The results for all-cause mortality in double-blind trials were similar to those in all trials combined. There was little evidence of vaccine protection among elderly patients or adults with chronic illness in analyses of all trials (RR 1.04, 95% CI 0.78 1.38, for presumptive pneumococcal pneumonia; 0.89, 95% CI 0.69-1.14, for all cause pneumonia; and 1.00, 95% CI 0.87-1.14, for all-cause mortality). INTERPRETATION: Pneumococcal vaccination does not appear to be effective in preventing pneumonia, even in populations for whom the vaccine is currently recommended. PMID- 19124792 TI - Making early childhood count. PMID- 19124793 TI - Painful swelling in the thigh: diabetic muscle infarction. PMID- 19124794 TI - Migratory pulmonary infiltrates. Goodpasture syndrome. PMID- 19124796 TI - Use administrative databases with caution. PMID- 19124800 TI - Facial swelling after a dental procedure. PMID- 19124801 TI - Doctors don't own medical knowledge. PMID- 19124802 TI - KRAS mutations and sensitivity to epidermal growth factor receptor inhibitors in colorectal cancer: practical application of patient selection. AB - Recent retrospective evidence from several randomized studies has established that advanced colorectal cancer patients with tumors harboring a mutation in the KRAS gene do not derive benefit from the administration of epidermal growth factor receptor-directed monoclonal antibodies, such as cetuximab or panitumumab. This represents a paradigm-changing event and will have substantial impact on current and future anticancer drug development. These results add to the economic and ethical considerations involved in the development of novel targeted therapies and should increase our scrutiny of mechanisms of resistance and predictive biomarkers while in earlier developmental stages. In this article we will review the available clinical data, discuss the potential implications for future drug development in colorectal cancer, and provide a comprehensive overview of the technical aspects of KRAS mutation testing. In particular we aimed at enumerating the available procedures for mutation detection and their main characteristics, as well as comparing them from a clinical feasibility standpoint. While the true specificity and sensitivity of these methods have yet to be fully characterized, a better understanding of the differences between tests will be critical so that clinicians and pathologists can fully integrate this testing into the routine care of patients with colorectal cancer. PMID- 19124803 TI - Evidence for cure by adjuvant therapy in colon cancer: observations based on individual patient data from 20,898 patients on 18 randomized trials. AB - PURPOSE: Limited data are available on the time course of treatment failures (recurrence and/or death), the nature and duration of adjuvant treatment benefit, and long-term recurrence rates in patients with resected stage II and III colon cancer. METHODS: The data set assembled by the Adjuvant Colon Cancer Endpoints Group, a collection of individual patient data from 18 trials and more than 20,800 patients testing fluorouracil-based adjuvant therapy in patients with stage II or III colon cancer, was analyzed. RESULTS: A significant overall survival (OS) benefit of adjuvant therapy was consistent over the 8-year follow up period. The risk of recurrence in patients treated with adjuvant chemotherapy never exceeds that of control patients, signifying that adjuvant therapy cures some patients, as opposed to delaying recurrence. After 5 years, recurrence rates were less than 1.5% per year, and after 8 years, they were less than 0.5% per year. Significant disease-free survival (DFS) benefit from adjuvant chemotherapy was observed in the first 2 years. After 2 years, DFS rates in treated and control patients were not significantly different, and after 4 years, no trend toward benefit was demonstrated. This benefit was primarily driven by patients with stage III disease. CONCLUSION: Adjuvant chemotherapy provides significant DFS benefit, primarily by reducing the recurrence rate, within the first 2 years of adjuvant therapy with some benefit in years 3 to 4, translating into long-term OS benefit. This reflects the curative role of chemotherapy in the adjuvant setting. After 5 years, recurrence rates in patients treated on clinical trials are low, and after 8 years, they are minimal; thus, long-term follow-up for recurrence is of little value. PMID- 19124804 TI - Rel a is an independent biomarker of clinical outcome in chronic lymphocytic leukemia. AB - PURPOSE: We recently demonstrated the biologic importance of the nuclear factor kappa B (NF-kappaB) subunit Rel A in chronic lymphocytic leukemia (CLL) and hypothesized that Rel A DNA binding would have prognostic significance in this disease. PATIENTS AND METHODS: Rel A DNA binding was quantified in nuclear extracts derived from 131 unselected CLL patient samples using a quantitative DNA binding enzyme-linked immunosorbent assay-based method. We then investigated the ability of Rel A to predict for the requirement for treatment and survival and compared our findings with other established prognostic markers. RESULTS: Rel A DNA binding was strongly associated with advanced Binet stage (P < .0001) but did not correlate with immunoglobulin V(H) (IgV(H)) mutation status (P = .25), CD38 expression (P = .87), or zeta-chain-associated protein kinase 70 (ZAP-70) expression (P = .55). It was predictive of time to first treatment (P = .02) and time to subsequent treatment (P = .0001). In addition, Rel A was the most predictive marker of survival both from date of diagnosis (hazard ratio [HR], 9.1; P = .01) and date of entry into the study (HR, 3.9; P = .05) and retained prognostic significance in multivariate analysis for both time to first treatment and overall survival in the presence of Binet stage, IgV(H) mutation status, CD38, and ZAP-70. CONCLUSION: Rel A is an independent prognostic marker of survival in CLL and seems to have the unique capacity to predict the duration of response to therapy. Prospective assessment of Rel A as a marker of clinical outcome and as a therapeutic target are now warranted. PMID- 19124805 TI - Pediatric-inspired therapy in adults with Philadelphia chromosome-negative acute lymphoblastic leukemia: the GRAALL-2003 study. AB - PURPOSE: Retrospective comparisons have suggested that adolescents or teenagers with acute lymphoblastic leukemia (ALL) benefit from pediatric rather than adult chemotherapy regimens. Thus, the aim of the present phase II study was to test a pediatric-inspired treatment, including intensified doses of nonmyelotoxic drugs, such as prednisone, vincristine, or L-asparaginase, in adult patients with ALL up to the age of 60 years. PATIENTS AND METHODS: Between 2003 and 2005, 225 adult patients (median age, 31 years; range, 15 to 60 years) with Philadelphia chromosome-negative ALL were enrolled onto the Group for Research on Adult Acute Lymphoblastic Leukemia 2003 protocol, which included several pediatric options. Some adult options, such as allogeneic stem-cell transplantation for patients with high-risk ALL, were nevertheless retained. RESULTS: were retrospectively compared with the historical France-Belgium Group for Lymphoblastic Acute Leukemia in Adults 94 (LALA-94) trial experience in 712 patients age 15 to 55 years. Results Complete remission rate was 93.5%. At 42 months, event-free survival (EFS) and overall survival (OS) rates were 55% (95% CI, 48% to 52%) and 60% (95% CI, 53% to 66%), respectively. Age remained an important bad prognostic factor, with 45 years of age as best cutoff. In older versus younger patients, there was a higher cumulative incidence of chemotherapy-related deaths (23% v 5%, respectively; P < .001) and deaths in first CR (22% v 5%, respectively; P < .001), whereas the incidence of relapse remained stable (30% v 32%, respectively). Complete remission rate (P = .02), EFS (P < .001), and OS (P < .001) compared favorably with the previous LALA-94 experience. CONCLUSION: These results suggest that pediatric-inspired therapy markedly improves the outcome of adult patients with ALL, at least until the age of 45 years. PMID- 19124806 TI - Accelerated telomere shortening precedes development of therapy-related myelodysplasia or acute myelogenous leukemia after autologous transplantation for lymphoma. AB - PURPOSE: Therapy-related myelodysplasia or acute myelogenous leukemia (t-MDS/AML) is a lethal complication of autologous hematopoietic stem-cell transplantation (aHCT) for Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL). Here, we investigated the hypothesis that accelerated telomere shortening after aHCT could contribute to the development of t-MDS/AML. PATIENTS AND METHODS: A prospective longitudinal cohort was constructed to investigate the sequence of cellular and molecular abnormalities leading to development of t-MDS/AML after aHCT for HL/NHL. This cohort formed the sampling frame for a nested case-control study to compare changes in telomere length in serial blood samples from patients who developed t-MDS/AML with matched controls who did not develop t-MDS/AML. RESULTS: An initial increase in telomere length at day 100 after aHCT was followed by an accelerated telomere shortening in t-MDS/AML patients when compared with controls. These telomere alterations preceded the onset of t-MDS and were independent of other known risk factors associated with development of t-MDS/AML on multivariate analysis. Additionally, we observed reduced generation of committed progenitors in patients who developed t-MDS/AML, indicating that these telomere alterations were associated with reduced regenerative capacity of hematopoietic stem cells. CONCLUSION: The development of t-MDS/AML after aHCT is associated with and preceded by markedly altered telomere dynamics in hematopoietic cells. Accelerated telomere loss in patients developing t-MDS/AML may reflect increased clonal proliferation and/or altered telomere regulation in premalignant cells. Genetic instability associated with shortened telomeres may contribute to leukemic transformation in t-MDS/AML. PMID- 19124807 TI - ABVD compared with BEACOPP compared with CEC for the initial treatment of patients with advanced Hodgkin's lymphoma: results from the HD2000 Gruppo Italiano per lo Studio dei Linfomi Trial. AB - PURPOSE: To compare doxorubicin, bleomycin, vinblastine, dacarbazine (ABVD) versus bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPP) versus cyclophosphamide, lomustine, vindesine, melphalan, prednisone, epidoxirubicin, vincristine, procarbazine, vinblastine, and bleomycin (COPPEBVCAD; CEC) for advanced Hodgkin's lymphoma (HL). PATIENTS AND METHODS: Three hundred seven patients with advanced HL (stage IIB, III, and IV) were randomly assigned to receive six courses of ABVD, four escalated plus two standard courses of BEACOPP, or six courses of CEC, plus a limited radiation therapy program. RESULTS: After a median follow-up of 41 months, BEACOPP resulted in a superior progression-free survival (PFS), with a significant reduction in risk of progression (hazard ratio [HR] = 0.50) compared with ABVD. No differences between BEACOPP and CEC, or CEC and ABVD were observed. The 5-year PFS was 68% (95% CI, 56% to 78%), 81% (95% CI, 70% to 89%), and 78% (95% CI, 68% to 86%), for ABVD, BEACOPP, and CEC, respectively (BEACOPP v ABVD, P = .038; CEC v ABVD and BEACOPP v CEC, P = not significant [NS]). The 5-year overall survival was 84% (95% CI, 69% to 92%), 92% (95% CI, 84% to 96%), and 91% (95% CI, 81% to 96%) for ABVD, BEACOPP, and CEC, respectively (P = NS). BEACOPP and CEC resulted in higher rates of grade 3-4 neutropenia than ABVD (P = .016); BEACOPP was associated with higher rates of severe infections than ABVD and CEC (P = .003). CONCLUSION: As adopted in this study BEACOPP is associated with a significantly improved PFS compared with ABVD, with a predictable higher acute toxicity. PMID- 19124808 TI - Are cancer-related decision aids effective? A systematic review and meta analysis. AB - PURPOSE: Decision aids (DAs) have been developed to improve communication between health professionals and patients, and to involve patients in decisions about their health care. Cancer-related decisions can be difficult due to problems in communicating complex information about prognosis and the modest benefits of available treatments. We conducted a systematic review of cancer-related DAs. METHODS: Randomized controlled trials (RCTs) of cancer-related DAs about screening, prevention, and treatment decision making were included. We completed a comprehensive literature search and conducted both qualitative and quantitative analyses. We also conducted a meta regression to explore heterogeneity of effect estimates. RESULTS: We identified 34 RCTs of DAs in a screening (n = 22 trials) or preventive/treatment (n = 12 trials) context. DAs significantly improved knowledge about screening options when compared to usual practice (weighted average effect size, 0.50; 95% CI, 0.27 to 0.73; P < .0001). A similar effect on knowledge was also found for preventive/treatment options (weighted average effect size, 0.50; 95% CI, 0.31 to 0.70; P < .0001). Overall, general anxiety was not increased in most trials and was significantly reduced in a screening context. Decisional conflict was reduced overall but not when screening and preventive/treatment studies were analyzed separately. There were few differences between different types of DAs. CONCLUSION: Cancer-related DAs are effective in increasing patient knowledge compared with usual practice without increasing anxiety particularly in the area of cancer screening. Further research is needed to determine the effectiveness of DAs in the prevention and treatment context. PMID- 19124809 TI - Cytogenetic profile predicts prognosis of patients with clear cell renal cell carcinoma. AB - PURPOSE: The majority of cytogenetic studies in renal cell carcinoma (RCC) have been impaired by small sample size, retrospective character, and lack of a survival end point. We prospectively studied the prognostic impact of cytogenetic abnormalities on a larger cohort of patients having up to 108 months of follow up. PATIENTS AND METHODS: Tumors of 282 patients who underwent nephrectomy for clear cell RCC were cytogenetically analyzed. Results were correlated with pathological factors and disease-specific survival. RESULTS: The most frequently observed cytogenetic abnormalities were loss of 3p (60%), gain of 5q (33%), loss of 14q (28%), trisomy 7 (26%), loss of 8p (20%), loss of 6q (17%), loss of 9p (16%), loss of 4p (13%), and loss of chromosome Y in men (55%). Tumors with loss of 3p presented at lower TNM stages. Loss of 4p, 9p, and 14q were all associated with higher TNM stages, higher grade, and greater tumor size. A deletion of 3p was associated with better prognosis (P = .03), while loss of 4p (P < .001), loss of 9p (P < .01), and loss of 14q (P < .01) were each associated with worse prognosis. Loss of the Y chromosome led to improved progression-free survival in metastatic patients (P = .02). In multivariate analysis, loss of 9p was retained as an independent prognostic factor. CONCLUSION: This cytogenetic study serves as a proof of principal that genetic information, such as loss of chromosome 9, can be obtained from widely available technology, and can provide additional prognostic information to standard clinicopathologic variables. PMID- 19124810 TI - Bilateral renal lymphoma after paraneoplastic limbic encephalitis. PMID- 19124811 TI - National Institutes of Health Consensus Development Conference Statement: management of hepatitis B. PMID- 19124813 TI - Summaries for patients: Outcomes in patients who acquired pneumonia in various settings. PMID- 19124812 TI - Antiviral therapy for adults with chronic hepatitis B: a systematic review for a National Institutes of Health Consensus Development Conference. AB - BACKGROUND: Chronic hepatitis B infection can lead to liver failure, hepatocellular carcinoma, and death. PURPOSE: To evaluate the effectiveness of antiviral therapy for adults with chronic hepatitis B infection. DATA SOURCES: Randomized, controlled trials (RCTs) of interferon (alpha2b and pegylated alpha2a), lamivudine, adefovir, entecavir, and telbivudine published from 1990 to 2008. STUDY SELECTION: Randomized, controlled clinical trials of adults with chronic hepatitis B published in English after 1989 that reported death; incidence of hepatocellular carcinoma or liver failure; prevalence and incidence of cirrhosis; presence or seroconversion of hepatitis B e antigen (HBeAg) or surface antigen (HBsAg), viral load of hepatitis B virus DNA; aspartate aminotransferase and alanine aminotransferase (ALT) levels; or fibrosis scores after therapy with interferon-alpha2b, pegylated interferon-alpha2a, lamivudine, adefovir, entecavir, and telbivudine. DATA EXTRACTION: Data extracted with standard protocols to calculate risk difference for clinical outcomes, viral load, HBeAg and HBsAg, ALT, histologic scores, and adverse events. DATA SYNTHESIS: In 16 RCTs (4431 patients), drug treatment did not improve clinical outcomes of chronic hepatitis B infection, but the trials were underpowered. In 60 RCTs that examined intermediate outcomes, no single treatment improved all intermediate outcomes. Low-quality evidence suggested HBsAg clearance after interferon-alpha2b (2 RCTs; 211 patients). Moderate-quality evidence suggested ALT normalization at follow-up after treatment with adefovir (2 RCTs; 600 patients) and HBeAg loss with lamivudine (2 RCTs; 318 patients). With interferon alpha2b, moderate-quality evidence suggested HBeAg loss (3 RCTs; 351 patients), seroconversion (2 RCTs; 304 patients), and ALT normalization (2 RCTs; 131 patients). Pegylated interferon-alpha2a versus lamivudine improved HBeAg seroconversion (1 RCT; 814 patients) and ALT normalization (2 RCTs; 905 patients) off treatment. Pegylated interferon-alpha2a combined with lamivudine versus lamivudine improved HBeAg loss (1 RCT; 543 patients) and ALT normalization (2 RCTs; 905 patients). Adverse events during antiretroviral therapy occurred in more than 50% of patients but were not associated with increased treatment discontinuation. However, most studies excluded patients with hepatic or renal insufficiency or other serious comorbid conditions. LIMITATION: Marked heterogeneity in study samples, interventions, and measured outcomes preclude definitive conclusions. CONCLUSION: Evidence was insufficient to assess treatment effect on clinical outcomes or determine whether inconsistent improvements in selected intermediate measures are reliable surrogates. Future research is needed to provide evidence-based recommendations about optimal antiviral therapy in adults with chronic hepatitis B infection. PMID- 19124814 TI - In the clinic. Cellulitis and soft-tissue infections. AB - This issue provides a clinical overview of cellulitis and soft-tissue infections, focusing on prevention, diagnosis, treatment, practice improvement, and patient information. Readers can complete the accompanying CME quiz for 1.5 credits. Only ACP members and individual subscribers can access the electronic features of In the Clinic. Non-subscribers who wish to access this issue of In the Clinic can elect "Pay for View." Subscribers can receive 1.5 category 1 CME credits by completing the CME quiz that accompanies this issue of In the Clinic. The content of In the Clinic is drawn from the clinical information and education resources of the American College of Physicians (ACP), including PIER (Physicians' Information and Education Resource) and MKSAP (Medical Knowledge and Self Assessment Program). Annals of Internal Medicine editors develop In the Clinic from these primary sources in collaboration with the ACP's Medical Education and Publishing division and with assistance of science writers and physician writers. Editorial consultants from PIER and MKSAP provide expert review of the content. Readers who are interested in these primary resources for more detail can consult www.acponline.org, http://pier.acponline.org, and other resources referenced within each issue of In the Clinic. PMID- 19124815 TI - Association between statin use and risk for keratinocyte carcinoma in the veterans affairs topical tretinoin chemoprevention trial. AB - BACKGROUND: Recent evidence suggests that statins may prevent cancer. OBJECTIVE: To quantify the association between statin use and the occurrence of keratinocyte carcinoma in high-risk veterans. DESIGN: Cohort study. SETTING: 6 Veterans Affairs medical centers. PARTICIPANTS: 1037 participants of the Veterans Affairs Topical Tretinoin Chemoprevention Trial, a randomized, multicenter, double-blind, vehicle-controlled trial of topical tretinoin, 0.1%, for prevention of keratinocyte carcinoma conducted from November 1998 to November 2004. MEASUREMENTS: Time to first occurrence of keratinocyte carcinoma on the face or ears. Participants using a statin at randomization, according to the Veterans Affairs Pharmacy Benefits Management database, were considered exposed. Study dermatologists conducted physical examinations at baseline and every 6 months during follow-up. The association between statin use at randomization and the outcome was evaluated by using propensity score matching (n = 608) and Cox proportional hazards regression (n = 1037). RESULTS: Among the 1037 participants, 37% used a statin at randomization (n = 397) for a median duration of at least 900 days over a median follow-up of 3.5 years. In the propensity score-matched analysis, statin use at randomization was not associated with keratinocyte carcinoma (rate ratio, 0.92 [95% CI, 0.73 to 1.16]), a finding that was consistent with the estimates derived from the Cox proportional hazards regression (rate ratio, 0.84 [CI, 0.70 to 1.02]). LIMITATIONS: The extent of residual confounding is unknown, and the confidence bounds around the measures of association were wide. These data may not be generalizable to lower-risk populations. CONCLUSION: These data show no conclusive or consistent relationship between long-term statin use and risk for keratinocyte carcinoma. PMID- 19124816 TI - Outcomes of patients hospitalized with community-acquired, health care associated, and hospital-acquired pneumonia. AB - BACKGROUND: Traditionally, pneumonia has been classified as either community- or hospital-acquired. Although only limited data are available, health care associated pneumonia has been recently proposed as a new category of respiratory infection. "Health care-associated pneumonia" refers to pneumonia in patients who have recently been hospitalized, had hemodialysis, or received intravenous chemotherapy or reside in a nursing home or long-term care facility. OBJECTIVE: To ascertain the epidemiology and outcome of community-acquired, health care associated, and hospital-acquired pneumonia in adults hospitalized in internal medicine wards. DESIGN: Multicenter, prospective observational study. SETTING: 55 hospitals in Italy comprising 1941 beds. PATIENTS: 362 patients hospitalized with pneumonia during two 1-week surveillance periods. MEASUREMENTS: Cases of radiologically and clinically assessed pneumonia were classified as community acquired, health care-associated, or hospital-acquired and rates were compared. RESULTS: Of the 362 patients, 61.6% had community-acquired pneumonia, 24.9% had health care-associated pneumonia, and 13.5% had hospital-acquired pneumonia. Patients with health care-associated pneumonia had higher mean Sequential Organ Failure Assessment scores than did those with community-acquired pneumonia (3.0 vs. 2.0), were more frequently malnourished (11.1% vs. 4.5%, and had more frequent bilateral (34.4% vs. 19.7%) and multilobar (27.8% vs. 21.5%) involvement on a chest radiograph. Patients with health care-associated pneumonia also had higher fatality rates (17.8% [CI, 10.6% to 24.9%] vs. 6.7% [CI, 2.9% to 10.5%]) and longer mean hospital stay (18.7 days [CI, 15.9 to 21.5 days] vs. 14.7 days [CI, 13.4 to 15.9 days]). Logistic regression analysis revealed that depression of consciousness (odds ratio [OR], 3.2 [CI, 1.06 to 9.8]), leukopenia (OR, 6.2 [CI, 1.01 to 37.6]), and receipt of empirical antibiotic therapy not recommended by international guidelines (OR, 6.4 [CI, 2.3 to 17.6]) were independently associated with increased intrahospital mortality. LIMITATIONS: The number of patients with health care-associated pneumonia was relatively small. Microbiological investigations were not always homogeneous. The study included only patients with pneumonia that required hospitalization; results may not apply to patients treated as outpatients. CONCLUSION: Health care-associated pneumonia should be considered a distinct subset of pneumonia associated with more severe disease, longer hospital stay, and higher mortality rates. Physicians should differentiate between patients with health care-associated pneumonia and those with community-acquired pneumonia and provide more appropriate initial antibiotic therapy. PMID- 19124818 TI - Nonhospital health care-associated hepatitis B and C virus transmission: United States, 1998-2008. AB - In the United States, transmission of hepatitis B virus (HBV) and hepatitis C virus (HCV) from health care exposures has been considered uncommon. However, a review of outbreak information revealed 33 outbreaks in nonhospital health care settings in the past decade: 12 in outpatient clinics, 6 in hemodialysis centers, and 15 in long-term care facilities, resulting in 448 persons acquiring HBV or HCV infection. In each setting, the putative mechanism of infection was patient to-patient transmission through failure of health care personnel to adhere to fundamental principles of infection control and aseptic technique (for example, reuse of syringes or lancing devices). Difficult to detect and investigate, these recognized outbreaks indicate a wider and growing problem as health care is increasingly provided in outpatient settings in which infection control training and oversight may be inadequate. A comprehensive approach involving better viral hepatitis surveillance and case investigation, health care provider education and training, professional oversight, licensing, and public awareness is needed to ensure that patients are always afforded basic levels of protection against viral hepatitis transmission. PMID- 19124817 TI - Brief communication: radiographic contrast infusion and catecholamine release in patients with pheochromocytoma. AB - BACKGROUND: Contrast-enhanced computed tomography (CT) is useful for localizing pheochromocytoma. However, in patients with suspected pheochromocytoma, CT is often canceled or not performed because of the strong belief that intravenous contrast may induce hypertensive crisis. OBJECTIVE: To examine whether intravenous low-osmolar contrast administration during CT induces catecholamine release that increases blood pressure or heart rate. DESIGN: Prospective study. SETTING: Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland. PARTICIPANTS: 22 patients with pheochromocytoma (15 nonadrenal and 7 adrenal) and 8 unmatched control participants without pheochromocytoma. MEASUREMENTS: Plasma catecholamine levels, blood pressure, and heart rate. RESULTS: Plasma catecholamine levels within and between groups did not significantly differ before and after intravenous administration of low osmolar CT contrast. Patients with pheochromocytoma experienced a clinically and statistically significant increase in diastolic blood pressure that was not accompanied by corresponding increases in plasma catecholamine levels. The difference became non-statistically significant after adjustment for use of alpha and beta-blockers. LIMITATION: The study lacked a placebo group, and the sample was relatively small. CONCLUSION: Intravenous low-osmolar contrast-enhanced CT can safely be used in patients with pheochromocytoma who are not receiving alpha- or beta-blockers. FUNDING: Eunice Kennedy Shriver National Institute of Child Health and Development, National Institutes of Health. PMID- 19124819 TI - Recommended adult immunization schedule: United States, 2009*. PMID- 19124820 TI - Immunization policy development in the United States: the role of the Advisory Committee on Immunization Practices. AB - The Advisory Committee on Immunization Practices (ACIP) consists of 15 experts in immunization and related fields, selected by the Secretary of the U.S. Department of Health and Human Services, to provide advice and guidance on control of vaccine-preventable diseases. In its role as a federal advisory committee, the ACIP develops written recommendations, subject to approval of the Director of the Centers for Disease Control and Prevention, for administration of U.S. Food and Drug Administration-licensed vaccines to children, adolescents, and adults in the U.S. civilian population. On the basis of careful review of available scientific data, including disease morbidity and mortality in the general U.S. population and in specific risk groups, vaccine safety and efficacy, cost-effectiveness, and related factors, the ACIP recommends vaccines and age for vaccine administration, number of doses and dosing interval, and precautions and contraindications. The ACIP works closely with several liaison organizations, including the American College of Physicians, to develop immunization recommendations that are harmonized among key professional medical organizations in the United States. This report includes a description of the member composition of the ACIP, the degree to which Committee members are screened for conflicts of interest, the workgroups that gather information before full Committee consideration, and the process and types of evidence used to formulate recommendations. PMID- 19124821 TI - Immunization guidelines for adult patients: an annual update and a challenge. PMID- 19124822 TI - The debt. PMID- 19124823 TI - How can cost-effectiveness information help control unsustainable growth in U.S. health care spending? PMID- 19124824 TI - How can cost-effectiveness information help control unsustainable growth in U.S. health care spending? PMID- 19124825 TI - Empirical use of fluconazole in critically ill patients: good study, but what was the hold-up? PMID- 19124826 TI - How can cost-effectiveness information help control unsustainable growth in U.S. health care spending? PMID- 19124827 TI - Empirical use of fluconazole in critically ill patients: good study, but what was the hold-up? PMID- 19124828 TI - Internet-enabled thyroid hormone abuse. PMID- 19124829 TI - Do race and ethnicity play a substantial role in the quality of care that patients receive in the U.S. Health care system? PMID- 19124830 TI - Heterologous fat transplantation for the treatment of HIV-related facial lipoatrophy. PMID- 19124832 TI - Detuning CD8+ T lymphocytes by down-regulation of the activating receptor NKG2D: role of NKG2D ligands released by activated T cells. AB - NKG2D is an activating receptor expressed on CD8(+)alphabeta(+) T cells, gammadelta(+) T cells, natural killer (NK) cells, and some CD4(+) T cells. For a long time, the interaction of NKG2D with its ligands (NKG2DLs) MICA, MICB, and ULBP1-3 has been considered a mechanism for recognition and elimination of tumor, infected, or otherwise "stressed" cells. However, a new role for NKG2D as an immunoregulatory receptor is emerging. Here, we show that NKG2D is strongly down modulated on antigen-activated CD8(+) T cells but only if CD4(+) T cells are present. Down-modulation was caused by soluble factors produced by CD4(+) T cells, and in particular soluble NKG2DLs were found in the supernatants of antigen-activated T-cell cultures. MICB was the ligand released at higher levels when CD4(+) T cells were present in the cell cultures, suggesting that it could be the major player of NKG2D down-modulation. CD8(+) T cells expressing low levels of NKG2D had impaired effector functions, as evaluated by proliferation, cytokine production, and cytotoxicity assays after combined triggering of NKG2D and TCR-CD3 complex. These findings show that activated CD4(+) T cells expressing NKG2DLs can efficiently prevent NKG2D-mediated CD8(+) T-cell functions, and suggest that the NKG2D/NKG2DL interaction can regulate immune responses. PMID- 19124831 TI - A comparative analysis of conventional and pretargeted radioimmunotherapy of B cell lymphomas by targeting CD20, CD22, and HLA-DR singly and in combinations. AB - Relapsed B-cell lymphomas are currently incurable with conventional chemotherapy and radiation treatments. Radiolabeled antibodies directed against B-cell surface antigens have emerged as effective and safe therapies for relapsed lymphomas. We therefore investigated the potential utility of both directly radiolabeled 1F5 (anti-CD20), HD39 (anti-CD22), and Lym-1 (anti-DR) antibodies (Abs) and of pretargeted radioimmunotherapy (RIT) using Ab-streptavidin (SA) conjugates, followed by an N-acetylgalactosamine dendrimeric clearing agent and radiometal labeled DOTA-biotin, for treatment of lymphomas in mouse models using Ramos, Raji, and FL-18 human lymphoma xenografts. This study demonstrates the marked superiority of pretargeted RIT for each of the antigenic targets with more complete tumor regressions and longer mouse survival compared with conventional one-step RIT. The Ab-SA conjugate yielding the best tumor regression and progression-free survival after pretargeted RIT varied depending upon the lymphoma cell line used, with 1F5 Ab-SA and Lym-1 Ab-SA conjugates yielding the most promising results overall. Contrary to expectations, the best rates of mouse survival were obtained using optimal single Ab-SA conjugates rather than combinations of conjugates targeting different antigens. We hypothesize that clinical implementation of pretargeted RIT methods will provide a meaningful prolongation of survival for patients with relapsed lymphomas compared with currently available treatment strategies. PMID- 19124834 TI - Improved outcome of allogeneic bone marrow transplantation due to breastfeeding induced tolerance to maternal antigens. AB - Exposure of offspring to noninherited maternal antigens (NIMAs) during pregnancy may have an impact on transplantations performed later in life. Using a mouse model, we recently showed that bone marrow transplantation (BMT) from NIMA exposed offspring to the mother led to a reduction of graft-versus-host disease (GVHD). Since offspring can also be exposed to NIMAs by breastfeeding after birth, we tested whether breast milk could mediate the tolerogenic NIMA effect. We found that oral exposure to NIMAs by breastfeeding alone was sufficient to reduce GVHD, and that in utero exposure to NIMAs is required for maximum reduction of GVHD. The tolerogenic milk effects disappeared when donor mice were injected with CD25 monoclonal antibodies during the lactation period, suggesting a CD4(+)CD25(+) regulatory T cell-dependent mechanism. Our results suggest a previously unknown impact of breastfeeding on the outcome of transplantation. PMID- 19124833 TI - p21-activated kinase regulates mast cell degranulation via effects on calcium mobilization and cytoskeletal dynamics. AB - Mast cells are key participants in allergic diseases via activation of high affinity IgE receptors (FcepsilonRI) resulting in release of proinflammatory mediators. The biochemical pathways linking IgE activation to calcium influx and cytoskeletal changes required for intracellular granule release are incompletely understood. We demonstrate, genetically, that Pak1 is required for this process. In a passive cutaneous anaphylaxis experiment, W(sh)/W(sh) mast cell-deficient mice locally reconstituted with Pak1(-/-) bone marrow-derived mast cells (BMMCs) experienced strikingly decreased allergen-induced vascular permeability compared with controls. Consistent with the in vivo phenotype, Pak1(-/-) BMMCs exhibited a reduction in FcepsilonRI-induced degranulation. Further, Pak1(-/-) BMMCs demonstrated diminished calcium mobilization and altered depolymerization of cortical filamentous actin (F-actin) in response to FcepsilonRI stimulation. These data implicate Pak1 as an essential molecular target for modulating acute mast cell responses that contribute to allergic diseases. PMID- 19124835 TI - Brave new world. PMID- 19124836 TI - Acute painless shoulder weakness during high-intensity athletic training. AB - BACKGROUND: High-intensity repetitive athletic activities may predispose the brachial plexus to repetitive stretch, compression, and subsequent injury, although painless shoulder weakness is a rare event. PURPOSE: The physical examination and electrodiagnostic findings in a series of United States Navy special warfare trainees who presented with acute painless shoulder weakness are presented, along with subsequent treatment and return-to-duty timeline. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: From August 2005 to August 2006, a total of 11 of 212 (5%) Navy Basic Underwater Demolition School trainees were identified with acute onset (<3 weeks) painless shoulder weakness without any prior shoulder injury. In all shoulders, symptoms began during a telephone pole lift-carry drill. All trainees underwent serial examinations, electrodiagnostic testing, and a comprehensive rehabilitation program. RESULTS: Physical examination revealed universal weakness in flexion and abduction and electrodiagnostic studies confirmed injury to the C5-6 area of the brachial plexus (axillary, suprascapular, and musculocutaneous). All 11 patients were removed from training and started on a physical therapy program until functional recovery at a mean of 21 weeks after onset of symptoms (range, 12-24). All 11 resumed military activities; however, only 6 completed the Navy Basic Underwater Demolition School program. CONCLUSION: In physically intense training or athletic environments, injuries to the upper brachial plexus may present with various forms of upper extremity dysfunction, including painless shoulder weakness. This information provides insight into a potentially debilitating shoulder problem and offers guidance on future training principles. PMID- 19124837 TI - College football player with unstable C1 fracture: a case report. PMID- 19124838 TI - A stereological approach for estimation of cellular immunogold labeling and its spatial distribution in oriented sections using the rotator. AB - Particulate gold labeling applied to ultrathin sections is a powerful approach for locating cellular proteins and lipids on thin sections of cellular structures and compartments. Effective quantitative methods now allow estimation of both density and distribution of gold labeling across aggregate organelles or compartment profiles. However, current methods generally use random sections of cells and tissues, and these do not readily present the information needed for spatial mapping of cellular quantities of gold label. Yet spatial mapping of gold particle labeling becomes important when cells are polarized or show internal organization or spatial shifts in protein/lipid localization. Here we have applied a stereological approach called the rotator to estimate cellular gold label and proportions of labeling over cellular compartments at specific locations related to a chosen cell axis or chosen cellular structures. This method could be used in cell biology for mapping cell components in studies of protein translocation, cell polarity, cell cycle stages, or component cell types in tissues. PMID- 19124839 TI - Localization of runx2, osterix, and osteopontin in tooth root formation in rat molars. AB - Cementogenesis starts with the differentiation of cementoblasts. Mature cementoblasts secrete cementum matrix. Cementum components are similar to bone; moreover, cementoblasts possess many characteristics similar to those of osteoblasts. Runx2 and osterix, the transcriptional factors for osteoblast differentiation, participate in tooth formation. However, the characteristics of Runx2 and osterix during the differentiation process of cementoblasts remain unclear. In this study, we examined the immunolocalization patterns of Runx2, osterix, and osteopontin during rat molar tooth formation. Periodontal ligament cells and osteoblasts located on the alveolar bone surface showed immunoreactivity for Runx2. Colocalization of Runx2 and osterix was detected in cementoblasts, which penetrated the ruptured Hertwig's epithelial root sheath and attached to root dentin. Moreover, osteopontin was observed in Runx2-positive cementoblasts facing the root surface. However, the cells adjacent to cementoblasts showed only Runx2 reactivity. Neither Runx2 nor osterix was seen in cementocytes. These results suggest that both Runx2 and osterix are important for differentiation into cementoblasts. Additionally, osterix may be indispensable for transcription of osteopontin expression. PMID- 19124841 TI - 3D visualization and measurement of capillaries supplying metabolically different fiber types in the rat extensor digitorum longus muscle during denervation and reinnervation. AB - The aim of this study was to determine whether capillarity in the denervated and reinnervated rat extensor digitorum longus muscle (EDL) is scaled by muscle fiber oxidative potential. We visualized capillaries adjacent to a metabolically defined fiber type and estimated capillarity of fibers with very high oxidative potential (O) vs fibers with very low oxidative potential (G). Capillaries and muscle fiber types were shown by a combined triple immunofluorescent technique and the histochemical method for NADH-tetrazolium reductase. Stacks of images were captured by a confocal microscope. Applying the Ellipse program, fibers were outlined, and the diameter, perimeter, cross-sectional area, length, surface area, and volume within the stack were calculated for both fiber types. Using the Tracer plug-in module, capillaries were traced within the three-dimensional (3D) volume, the length of capillaries adjacent to individual muscle fibers was measured, and the capillary length per fiber length (Lcap/Lfib), surface area (Lcap/Sfib), and volume (Lcap/Vfib) were calculated. Furthermore, capillaries and fibers of both types were visualized in 3D. In all experimental groups, O and G fibers significantly differed in girth, Lcap/Sfib, and Lcap/Vfib, but not in Lcap/Lfib. We conclude that capillarity in the EDL is scaled by muscle fiber size and not by muscle fiber oxidative potential. PMID- 19124840 TI - Region-specific differentiation potential of adult rat spinal cord neural stem/precursors and their plasticity in response to in vitro manipulation. AB - This study characterized the differentiation of neural stem/precursor cells (NSPCs) isolated from different levels of the spinal cord (cervical vs lumbar cord) and different regions along the neuraxis (brain vs cervical spinal cord) of adult male Wistar enhanced green fluorescent protein rats. The differentiation of cervical spinal cord NSPCs was further examined after variation of time in culture, addition of growth factors, and changes in cell matrix and serum concentration. Brain NSPCs did not differ from cervical cord NSPCs in the percentages of neurons, astrocytes, or oligodendrocytes but produced 26.9% less radial glia. Lumbar cord NSPCs produced 30.8% fewer radial glia and 6.9% more neurons compared with cervical cord NSPCs. Spinal cord NSPC differentiation was amenable to manipulation by growth factors and changes in in vitro conditions. This is the first study to directly compare the effect of growth factors, culturing time, serum concentration, and cell matrix on rat spinal cord NSPCs isolated, propagated, and differentiated under identical conditions. PMID- 19124842 TI - Differentiation-dependent PTPIP51 expression in human skeletal muscle cell culture. AB - Protein tyrosine phosphatase-interacting protein 51 (PTPIP51) expression was analyzed in proliferating and differentiating human myogenic cells cultured in vitro. Satellite cell cultures derived from four different individuals were used in this study. To analyze the expression of PTPIP51, myoblasts were cultured under conditions promoting either proliferation or differentiation. In addition, further differentiation of already-differentiated myobtubes was inhibited by resubmitting the cells to conditions promoting proliferation. PTPIP51 protein and mRNA were investigated in samples taken at defined time intervals by immunostaining, immunoblotting, in situ hybridization, and PCR. Image analyses of fluorescence immunostainings were used to quantify PTPIP51 in cultured myoblasts and myotubes. Myoblasts grown in the presence of epidermal and fibroblast growth factors (EGF and FGF), both promoting proliferation, expressed PTPIP51 on a basic level. Differentiation to multinuclear myotubes displayed a linear increase in PTPIP51 expression. The rise in PTPIP51 protein was paralleled by an augmented expression of muscle-specific proteins, namely, sarcoplasmic reticulum Ca(2+) ATPase and myosin heavy-chain protein, both linked to a progressive state of myotubal differentiation. This differentiation-induced increase in PTPIP51 was partly reversible by resubmission of differentiated myotubes to conditions boosting proliferation. The results clearly point toward a strong association between PTPIP51 expression and differentiation in human muscle cells. PMID- 19124843 TI - Genome-wide association study of plasma lipoprotein(a) levels identifies multiple genes on chromosome 6q. AB - Plasma lipoprotein(a) (Lp[a]) level is an independent risk factor of cardiovascular disease that is under strong genetic control. We conducted a genome-wide association study of plasma Lp(a) in 386 members of a founder population that adheres to a communal lifestyle, proscribes cigarette smoking, and prepares and eats meals communally. We identified associations with 77 single nucleotide polymorphisms (SNPs) spanning 12.5 Mb on chromosome 6q26-q27 that met criteria for genome-wide significance (P 160 and >170 beats/min for 85.3% and 46.7% of racing, respectively. Sweat rates were 1.06 +/- 0.12 L/h or 13.4 +/- 1.2 mL . kg-1 . h-1, and postrace dehydration was 0.6% +/- 0.6% of prerace body mass. Drivers rated thermal sensation as hot (10.3 +/- 0.9), thermal discomfort as uncomfortable (3.1 +/- 1.0), and perceived exertion as very hard to very, very hard (8.7 +/- 1.7) after the races. Overall physiological and perceptual strain were 7.4 +/- 1.0 and 7.1 +/- 1.2, respectively. CONCLUSIONS: Despite the use of cooling, V8 Supercar drivers endure thermal, cardiovascular, and perceptual strain during brief driving bouts in hot conditions. PMID- 19124906 TI - Diurnal variation in swim performance remains, irrespective of training once or twice daily. AB - PURPOSE: Fast swim times in morning rounds are essential to ensure qualification in evening finals. A significant time-of-day effect in swimming performance has consistently been observed, although physical activity early in the day has been postulated to reduce this effect. The aim of this study was to compare intradaily variation in race-pace performance of swimmers routinely undertaking morning and evening training (MEG) with those routinely undertaking evening training only (EOG). METHODS: Each group consisted of 8 swimmers (mean +/- SD: age = 15.2 +/- 1.0 and 15.4 +/- 1.4 y, 200-m freestyle time 132.8 +/- 8.4 and 136.3 +/- 9.1 s) who completed morning and evening trials in a randomized order with 48 h in between on 2 separate occasions. Oral temperature, heart rate, and blood lactate were assessed at rest, after a warm-up, after a 150-m race-pace swim, and after a 100-m time trial. Stroke rate, stroke count, and time were recorded for each length of the 150-m and 100-m swims. RESULTS: Both training groups recorded significantly slower morning 100-m performances (MEG = +1.7 s, EOG = +1.4 s; P < .05) along with persistently lower morning temperatures that on average were 0.47 degrees C and -0.60 degrees C, respectively (P < .05). No differences were found in blood-lactate, heart-rate, and stroke-count responses (P > .05). All results were found to be reproducible (P > .05). CONCLUSIONS: The long-term use of morning training does not appear to significantly reduce intradaily variation in race-pace swimming or body temperature. PMID- 19124907 TI - Effects of varying post-warm-up recovery time on 200-m time-trial swim performance. AB - CONTEXT: Warm-up before athletic competition might enhance performance by affecting various physiological parameters. There are few quantitative data available on physiological responses to the warm-up, and the data that have been reported are inconclusive. Similarly, it has been suggested that varying the recovery period after a standardized warm-up might affect subsequent performance. PURPOSE: To determine the effects of varying post-warm-up recovery time on a subsequent 200-m swimming time trial. METHODS: Ten national-caliber swimmers (5 male, 5 female) each swam a 1500-m warm-up and performed a 200-m time trial of their specialty stroke after either 10 or 45 min of passive recovery. Subjects completed 1 time trial in each condition separated by 1 wk in a counterbalanced order. Blood lactate and heart rate were measured immediately after warm-up and 3 min before, immediately after, and 3 min after the time trial. Rating of perceived exertion was measured immediately after the warm-up and time trial. RESULTS: Time-trial performance was significantly improved after 10 min as opposed to 45 min recovery (136.80 +/- 20.38 s vs 138.69 +/- 20.32 s, P < .05). There were no significant differences between conditions for heart rate and blood lactate after the warm-up. Pre-time-trial heart rate, however, was higher in the 10-min than in the 45-min rest condition (109 +/- 14 beats/min vs 94 +/- 21 beats/min, P < .05). CONCLUSIONS: A post-warm-up recovery time of 10 min rather than 45 min is more beneficial to 200-m swimming time-trial performance. PMID- 19124908 TI - The use of static stretching in warm-up for training and competition. AB - Static stretching (SS) is widely used in warm-ups before training and competition. A growing amount of research, however, has demonstrated that SS can impair muscle performance, leading to a reevaluation of optimal warm-up protocols. This commentary discusses many of the methodological issues that can influence conclusions about the acute effects of SS on performance. One difficulty in interpreting the literature is the lack of control or communication about the volume and intensity of the various stretching treatments used. Another major issue is the failure of many researchers to evaluate SS as it is used in practice, particularly the interaction with the other general and sport-specific components of the warm-up. Acute warm-up effects on performance should be considered in conjunction with potential effects on injury prevention. Future directions in research include optimizing general and sport-specific warm-ups, time course of physiological and performance effects, and individualization of warm-ups according to fitness and skill level. PMID- 19124909 TI - TB/HIV co infection in Georgia. AB - HIV and TB co-infection is a considerable problem worldwide. HIV significantly increases the morbidity and mortality from TB and often makes the diagnosis more challenging. In this study we attempted to evaluate the prevalence of TB among newly diagnosed HIV infected persons and determine the rate of HIV infection among active TB patients in Georgia. The prospective observational study has been conducted in Georgia since January 01, 2006. All newly diagnosed HIV positive persons were screened for active and latent TB infection and the prevalence of TB was identified. During the same time period HIV screening was performed in all identified active TB cases. Up to 22% (16.7 to 22%) of HIV positive individuals were found to have active TB, and 22.4 to 32.6% had LTBI. The prevalence of HIV among TB patents ranged from 1.7 to 2.2%. The study showed significant prevalence of TB (both active and latent TB) among HIV patients. Because of problems with TB diagnosis in HIV patients, the real prevalence may be underestimated. The alarming statistical data should force us towards meticulous and scrupulous screening for tuberculosis among HIV positive individuals. The prevalence of HIV among TB patents was not very high, ranging from 1.7 to 2.2%, but we recommend routine screening for HIV in all active TB patients. PMID- 19124910 TI - Antiretroviral treatment in Georgia. AB - HIV infection is the major public health, social and economic problem in Georgia. The aim of this study is to evaluate effectiveness of ARV treatment system in Georgia. Study included 1052 people living with HIV/AIDS in Georgia registered at Infectious Disease, AIDS and Clinical Immunology Research Center since 2004. To ensure universal access to ARV therapy all HIV/AIDS individuals included in the study were investigated by special algorithm, all identified patients requiring ARV therapy were offered treatment and monitored during therapy on treatment effectiveness and side effects. Detection of HIV antibodies was performed by ELISA with further confirmation by Western Blot Assay. HIV-1 RNA in plasma was measured by quantitative Polymerase Chain Reaction. For determination of percentages and absolute count of T lymphocyte subpopulations single-platform immunophenotyping technique using the Becton-Dickinson FACSCalibur flow cytometer was applied. For resistance testing TRUGENE HIV-1 Genotyping Kit with the OpenGene DNA Sequencing System (Siemens) was used. Treatment was offered to 595 HIV/AIDS patients. 594 patients started treatment, 1 patient refused. Out of treated 594 HIV/AIDS patients 22 patients discontinued, 111 patients died and 461 patients are currently on ARV treatment. Out of treated patients 406 adults and 21 children are receiving first-line treatment, 31 adults and 2 children are on second-line treatment and 1 adult is receiving salvage regimen. Treatment failure was defined in 55 cases. Among them immunological failure was observed in 7 cases, clinical failure in 1 case and virologic failure in 47 cases. Prevalence of drug resistance among virologic failure cases accounted for 72% and inadequate adherence for 28% cases. Majority of death cases among ARV treated patients was due to non-AIDS related or incurable conditions, while deaths due to AIDS related conditions mainly were associated to the delayed referral of patients in already advanced stage of disease. It's worth to mention that highest number of death cases was due to liver failure in HIV/HCV and/or HBV co-infected patients. PMID- 19124911 TI - Successful application of laboratory tools for the detection of HIV drug resistance in routine clinical care in Georgia. AB - Since 2004, Georgia the first among Eastern European countries ensured universal access to highly active antiretroviral therapy (HAART). Laboratory monitoring of HAART using CD4 count, viral load (VL) and HIV genotypic resistance testing was carried out in according with National HIV/AIDS Treatment Guidelines. Georgia the first among former Soviet Union countries implemented HIV genotypic resistance testing in HIV clinical care. The present paper reports on successful application of laboratory tools in routine clinical care for the early detection of HIV drug resistance. For genotypic resistance testing the TruGene HIV-1 Genotyping Kit (Bayer HealthCare LLC, Tarrytown, NY) was used according to manufacturer's instructions. Analysis included 45 patients with virologic failure. Of them 34 (75.5%) had at least one resistant mutation. Dual-class drug resistance was found in 30 (66.7%) patients. One (2.2%) patient carried triple-class resistance mutations. Median number of resistant mutations was 2. Most commonly detected NRTI mutation was M184/V/I (68.9%). G190S/A was the most frequent NNRTI mutation (42.2%), followed by K103N (28.9%). All patients with drug resistance mutations were switched to a second line regimens. Analysis of virologic and immunological outcomes among 23 patients who had at least two follow-up measurements of CD4 and VL after resistance test, showed statistically significant decrease in VL by 2.5 log(10) and mean gain of 181 cells/mm(3) in CD4 count by the last available measurement. Routine monitoring of VL and subsequent use of HIV drug resistance testing allowed for early identification of HIV drug resistance, reducing the opportunity for mutations to accumulate. Routine use of sophisticated laboratory methods for HAART monitoring has beneficial impact on clinical outcomes and should be used as part of the strategy to combat resistance. PMID- 19124912 TI - Implementation of PMTCT in Georgia. AB - To review the existing experience in prevention of Mother-To-Child Transmission (PMTCT) of HIV in Georgia the comprehensive PMTCT state program was started in 2005. Georgia was the first among the former Soviet Countries that ensured the universal access to PMTCT throughout the Country. According to the National PMTCT protocol, all pregnant women are offered Voluntary Counseling and Testing for HIV infection at Women Health Centers, maternity hospitals, and regional hospitals of Georgia. Positive results are referred to the Infectious Diseases, AIDS and Clinical Immunology Research Center (IDACIRC) for the confirmation and management that implies: antiretroviral therapy, caesarean section, infant feeding by formula and PCP prophylaxis by TMP-CTX. Data were collected using National HIV/AIDS Data Base. Prevalence of HIV among pregnant women attending VCT services in 2005-2008 years was 0.03%. Throughout the period 1999-2008 total 84 pregnancies were registered at the IDACIRC, among them 77 pregnancies were monitored by IDACIRC. Prophylactic strategy was tailored individually according to the national acting guideline, women gestation age, HIV disease stage, ARV's availability, etc. Totally 36 pregnant women received full PMTCT service. In this group no vertical transmission of HIV infection was recorded. 33 pregnant women received partial PMTCT service. The reasons were: late HIV diagnosis, limited access to ARV (from 1999 till 2004), refusal by pregnant woman. Number of HIV transmission cases was 3 in this group. As of November, 2008 eight women are still pregnant. Since 2005 Georgia ensured comprehensive and sustainable PMTCT service throughout the Country and universal access for all pregnant women. Provision of full package of this service minimized the risk of vertical transmission. PMID- 19124913 TI - Distribution of HIV-1 resistant polymorphisms among HIV infected patients in Georgia. AB - Host genetic factors are believed to play an important role in the pathogenesis and natural history of HIV disease along with determining the rate and severity of HIV epidemic in a particular country. CCR5, CCR2 and SDF1 genes are known to influence the susceptibility to HIV-1 infection and to be involved in the rate of disease progression. Unlike CCR5 Delta32 mutation, mutations in CCR2-64I and SDF1 3A do not provide full protection against HIV-1 acquisition, however, they are believed to delay the onset of AIDS defining illness. The objectives of this study were to evaluate the prevalence of host genetic factors among HIV infected patients in Georgia in order to define the correlations between CCR5Delta32, CCR 64I and SDF1-3A genotypes and HIV disease progression in our country. 120 HIV infected individuals were enrolled in the study. Mutations were detected by the polymerase chain reaction/restriction fragment length polymorphism method. We have studied the DNA polymorphisms at the loci that encode these proteins in 120 HIV infected individuals. As expected, no CCR5 homozygous 32 base pair mutation was found among HIV infected persons, however 6 heterozygous patients produced allele frequency 2.5%. Allele frequency of CCR2 and SDF1 allele was equal to 10.75% and 32% respectively. The overall frequency of CCR2 and CCR5 mutations is comparable to their frequency among European populations. However, to our knowledge, the frequency of SDF1-3A allele frequency in Georgians is higher than has been reported in European countries. We found a delay in the progression of HIV infection among persons who were between heterozygous for the CCR5 Delta32 mutation. In order to explore the impact of host genetic factors on the HIV epidemic in Georgia, host genetic studies involving different groups would be of interest. PMID- 19124914 TI - Neurological complications in patients with HIV/AIDS. AB - The aim of the study was to determine the prevalence of HIV-related neurological disorders in HIV positive patients and its relationship to CD4 cell counts in Georgia. This study included 388 HIV/AIDS patients (302 men and 86 women), who have been admitted to the in-patient Department of Infectious Diseases, AIDS and Clinical Immunology Research Center (IDACIRC) of Georgia since 2006. Diagnosis of neurological disorders was made based on clinical symptoms and instrumental laboratory investigations. CNS Neurological complications were detected in 76 patients; 13 patients had two or more neurological complications. Tuberculosis meningitis were the most common neurological disorders 26 (34%), followed by CNS toxoplasmosis 17 (22%), cryptococcal meningitis 11 (15%), presumed CMV encephalitis 5 (7%), PML 4 (5%), primary CNS lymphoma 4 (5%) and bacterial meningitis 3 (4%). AIDS related dementia was detected in 18 patients (24%). The median CD4+ T lymphocyte count was 47 cells/mm(3) (range: 2-183 cells/mm(3)) in HIV patients with neurological complications. There was correlation between the CD4 T lymphocyte count and type of neurological manifestation. Namely, in the patients with HIV related dementia median CD4 T lymphocyte count was 164 cells/mm(3), in the patients with CNS toxoplasmosis median CD4 count was 83 cells/mm(3), in the patients with cryptococcal meningitis median CD4 T lymphocyte count was 34 cells/mm(3) and in the patients with CMV encephalitis median CD4 T lymphocyte count was 26 cells/mm(3). Some neurological disorders such as TB meningitis and bacterial meningitis can occur at any CD4 level. PML and primary CNS lymphoma occurred when CD4 T lymphocyte count < 50 cells/mm(3). The most common clinical manifestations of neurological disorders in HIV infected patients were headache (91%), fever (75%), focal neurological deficits (61%), speech disturbances (42%), cognitive dysfunction (41%), visual disturbances (36%), impaired coordination (29%) and seizures (15%). The study provide convincing evidence that neurological disorders with HIV infection might serve as an indicator for advanced HIV infection, immunosuppression and decreased CD4 cell counts. Our data have shown correlation between the type of neurological manifestations of HIV infection and CD4 T lymphocyte count. PMID- 19124915 TI - Assessment of liver fibrosis and cirrhosis by transient elastography among patients with chronic HBV and HCV infection in Georgia. AB - The aim of the study was to evaluate liver fibrosis (LF) and cirrhosis using Transient elastography (TE) using Fibroscan in patients with chronic HBV and HCV infection. The device evaluates LF by measurement of liver stiffness (LS). 525 patients with chronic HCV infection and 105 patients with chronic HBV infection were included in the study. These patients were investigated at the Georgian French joint hepatology clinic "Hepa" from November 2007, till November 2008. Among investigated HBV infected 105 patients 65 (61.9%) had no fibrosis (LS<5.5 kpa), 23 (21.9%) had mild fibrosis (LS-5.5-8.0 kpa), 9 (8.6%) had severe fibrosis (LS-8.0-14.0 kpa) and 8 (7.6%) had liver cirrhosis (LS>14.0 kpa). Among investigated HCV infected 525 patients 200 (38.1%) had no fibrosis (LS<5.5 kpa), 139 (26.5%) patients had mild fibrosis (LS-5.5-8.0 kpa), 87 (16.5%) patients had severe fibrosis (LS-8.0-14.0 kpa) and 99 (18.9%) patients had liver cirrhosis. It is concluded that transient elastography (TE) using Fibroscan is simple, non invasive, reliable and easily reproducible method for assessing liver fibrosis and cirrhosis in patients with chronic HBV and HCV infection. TE is characterized with an excellent accuracy. TE results are well correlated with the clinical signs as well as with the results of laboratory and instrumental investigations. Fibrosis stages by Metavir measured using Fibroscan well corresponds with the liver biopsy results. Considering the high prevalence of fibrosis and cirrhosis among patients with chronic HBV and HCV infection, TE is a very valuable method for detecting early stages of fibrosis allowing to avoid the progression of liver damage, as well as end-stage liver disease. TE is easy to perform and therefore allows regular follow-up of the course of LF. PMID- 19124916 TI - Acute/recent HCV infection. Clinical course, viral replication kunetic and disease outcome. AB - The aim of the study was to reveal and investigate acute/recent HCV infection at the very early stage in seronegative blood donors and seronegative Injecting Drug Users (IDUs) and to assess clinical laboratory variants of infection, viral replication kinetic, disease outcome, host and viral characteristics. Two groups of patients were included in this study. The first group consisted of ELISA negative 7000 blood donors; the second group included 3000 Injecting Drug Users (IDUs). All patients were investigated on HCV RNA by qualitative PCR using mini pool method. A pool of 6 was applied for blood donors' and a pool of 5 for IDUs. PCR negative pools were excluded from the study, while PCR positives were examined on individual samples. Anti-HCV was detected by ELISA and RIBA. Detection HCV RNA was performed by Real time PCR technique using COBAS TaqMan Test. HCV genotyping--by INNO-Lipa. HLA typing--by Sequence Specific Primer Amplification (SSP). 16 patients with acute/recent HCV were revealed: 7 from blood donors, 9 from IDUs. Among them: 4 were symptomatics and 12 asymptomatics. Out of 4 symptomatics 3 were with jaundice. Among 12 asymptomatics: 8 had elevated ALT; 2 neither elevated ALT nor symptoms but developed anti-HCV; 2 were with normal ALT and without further anti-HCV seroconversion. Among 16 subjects: 9 had genotype -1b, 1--genotype 1a, 3--genotype 2a/2c and 3--genotype 3a. Out of 16 cases 4 cleared the virus; 12 developed chronic infection. Spontaneous clearance (recovery from the disease) was observed in 2 out of 4 symptomatic patients and only in 2 patients out of 12 asymptomatics. In all patients viremia increased rapidly and reached a peak by week 4. Viral titer was remarkably stable for the next three weeks, followed by two or three fold decrease by week 9. After week 10 the viremia rapidly decreased: 4 or 5 logs by week 12 and it became either undetectable by weeks 16-18 (viral clearance), or virus was not eliminated and viral titer persisted in all follow up period (chronic infection). HLA DRB1 1101, DQB1 0301 and DRB1 1301/DQA1 0103 alleles were associated with clearance of HCV whereas DRB1 0301 was associated with chronic infection. Prevalence of HCV among seronegative blood donors was 0.1% and among IDUs 0.3%. Among acute/recent HCV infected patients rate of chronicity was 75% (50% in symptomatics and 83% in asymptomatics). Rate of recovery was 50% in symptomatic patients and about 16% in asymptomatics. Acute/recent HCV infection might have following clinical laboratory forms: symptomatic disease with or without jaundice, asymptomatic with or without elevated ALT, but with further anti-HCV seroconversion. It remains unclear whether enigmatic form of disease--acute/recent HCV infection without further seroconversion exists or not. PMID- 19124917 TI - Important aspects of nosocomial bacterial resistance and its management. AB - The article reviews management of nosocomial bacterial resistance aspects. Nosocomial infections are associated with substantial morbidity, mortality and cost. During the last several decades multi-drug resistant organisms increased in number considerably. Methicillin-resistant staphylococcus aureus, Vancomycin intermediately resistant staphylococcus aureus and fully vancomycin-resistant staphylococcus aureus evolved as a consequence of methicillin and vancomycin use. The introduction of third generation cephalosporins were followed by emergence of extended spectrum and AMP-C -lactamases among gram negative bacteria, and carbapenems were targeted by carbapenemases. The poor diagnostic yield of current microbiologic methods in identifying certain resistant organisms, combined with decreasing numbers of newly developed antibiotics pose a significant challenge to physicians. We reviewed some of the approaches which can be followed to maximize the positive clinical outcome in patients with resistant nosocomial infections, using currently available antibiotics. More sensitive microbiological methods and new types of antibiotics are needed to adequately address the problem in the future. PMID- 19124918 TI - Prevalence of hepatitis B and C among HIV positive patients in Georgia and its associated risk factors. AB - The aim of the study was to determine the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) co-infection among HIV positive patients, to identify most relevant risk factors of co-infection and develop preventive interventions. Study participants were voluntary individuals 18 years of age or older recruited from AIDS Center VCT unit in Tbilisi, Georgia. Eligibility criteria of participants were: HIV positive result confirmed by western blot; age; and voluntary participation. Total 175 patients undergo interview with specially designed questionnaires. Most of the participants were male (71.4%), age range of HIV positives varied from 20 to 77 years old. Prevalence of HCV among HIV positive patients is high. Almost half (48.57%) HIV positive patients are co infected with HCV. Men were more likely than women co-infected with HCV (60.80% and 18% accordingly). Major risk factor of male co-infection was related to drug use, needle and injection equipment sharing. Prevalence of HCV among injecting drug users was (73.40%). Drug users had 3.25 times more risk (PR 3.25; 95%CI; CL- 1.89-5.26; p<0.01) to be infected with HCV compare non IDUs. Prevalence of being infected with HBV (Anti-HBc) among HIV positives was 43.42% (76/175) and the prevalence of Chronic HBV (HBsAg positive) was 6.86% (12/175). Prevalence rate of HBsAg among IDUs was 8.51% and among non IDU participants 5.26%. Triple infection (HIV, Hepatitis C and chronic form of Hepatitis B--HBsAg) was among 9 patients (5.14%). Infections were associated with injection drug use (88.88%) and mostly were related to share of needles/syringes and other injecting medical equipment. Transmission of HBV and HCV by sexual contact was not observed among those 9 participants. High risk behavior among HIV positive participants mostly related to drug use and unprotected sex with non regular partners. Other risk factors for Hepatitis transmission were associated with invasive medical manipulations, blood transfusion, surgery, abortions and etc. None of cases of HIV, or Hepatitis (B, C) transmission through medical manipulations can be documentary proved based on those research data. PMID- 19124919 TI - Oral lesions in HIV-positive patients in Georgia. AB - To study the prevalence of oral lesions in HIV infected patients and its relationship with CD4+ cell count in Georgia 732 HIV positive adult patients who were admitted to the Infectious Diseases, AIDS and Clinical Immunology Research Center (IDACIRC) since January, 2006 till October, 2008 were evaluated. Each patient underwent full clinical and standard laboratory examination. CD4+ cell count was determined by the Becton-Dickinson FACSCalibur flow cytometer (MultiTEST CD3 FITC/CD8 PE/CD45 PerCP/CD4 APC Reagent). Socio-demographic data was obtained using a standard questionnaire at the epidemiology department of IDACIRC. Oral manifestations were diagnosed according to EEC clearinghouse classification (1993). Oral lesions were revealed in 546 patients (75%). 186 patients (25%) did not exhibit any oral complications. The prevalence of two or more simultaneously exhibited types of lesions was as follows: three types of lesions were detected in 45 patients (6%) and two types of lesions were detected in 245 patients (33%). The investigation revealed oral candidiasis constituted the most common form of oral lesions, representing a 64% (467 patients), followed by HIV associated periodontal diseases in 216 patients (30%), recurrent aphthous like ulcerations in 118 patients (16%), oral hairy leukoplakia in 58 patients (8%), orolabial herpes simplex infection in 50 patients (7%), human papillomavirus (wart like lesions) in 37 patients (5%) and Kaposi's sarcoma in 3 patients (0.4%). Most of oral lesions cases were found in patients with low CD4+ cell count. Results of this study provide evidence that mucous membrane disorders with HIV infection might serve as an indicator for advanced HIV infection, immunosuppression and decreased CD4 cell counts. The physicians who are taking care of HIV patients have to be familiar with HIV-associated mucocutaneous diseases, their diagnoses, and management. PMID- 19124920 TI - World Health Organization's HIV/AIDS policy and Georgia. AB - WHO takes the lead within the UN system in the global health sector response to HIV/AIDS. Recent estimates indicate that the health sector alone represents at least 55% of the resources required for the global response to HIV/AIDS. In order to better target much-needed interventions, the WHO HIV/AIDS Programme focuses on five strategic directions: Enable people to know their HIV status; Maximize the health sector's contribution to HIV prevention; Accelerate the scale-up of HIV treatment and care; Strengthen and expand health systems; Invest in strategic information to guide a more effective response. The European Region is now experiencing the fastest rate of growth of HIV prevalence in any region of the word with the evidence of increasing transmission of HIV in several countries. In WHO/EURO was developed 13 new or revised evidence-based protocols on HIV/AIDS prevention, treatment and care including co-infection with hepatitis B and hepatitis C, with TB, and a protocol on sexual and reproductive health for people living with HIV/AIDS. WHO assisted Georgia to elaborate and implement its first 1994-95 National Workplan on HIV/AIDS. WHO/EURO activities in Georgia consist: The advocacy and promotion of HIV/AIDS-related WHO policies, strategies, recommendations, distribution of WHO publications, press releases, and reports; The technical assistance to MoLHSA, UN Theme Group on HIV/AIDS, GFATM, major national and international stakeholders involved in HIV/AIDS field in promotion, development and implementation of evidence based norms and standards. By the support of WHO seven guidelines and treatment protocols have been prepared and adopted on different topics of HIV/AIDS in Georgian language; WHO participated in the preparation of "Country Response Plans for HIV/AIDS ", "UN joint response plans for HIV/AIDS", "TB/HIV national plan", applications for GFATM proposals, UNGASS reports etc.; Support in promotion, planning and caring out HIV/AIDS related events (World AIDS Day, AIDS Memorial Day, etc). By the financial support of WHO/EURO in 2007-2008 was carried out the HIV/Hepatitis coinfection epidemiological study as well as in 2006 the socio-economic study for HIV/AIDS in Georgia; In different fields of HIV/AIDS (epidemiology, surveillance, diagnostic and treatment) around 20 National and Regional conferences and workshops have been organized in Georgia and about 400 Georgian specialists passed the training courses for capacity building in and outside country. PMID- 19124921 TI - Prevalence of HCV and genotypes distribution in general population of Georgia. AB - The aim of four years study was to determine the prevalence of HCV infection in the general population of Georgia and to assess HCV genotypes spread among them. For performing the planned investigation a cross-sectional study design was applied. Study subjects were Tbilisi residents selected through multiple clusters sampling technique application. Tbilisi is divided into ten districts. 2000 persons from the general adult population of Tbilisi, Georgia were enrolled in the study. The multi-stage cluster sampling method was applied for selection of study subjects. Districts of Tbilisi were considered as sub-populations. At the first stage the number of Population distribution by each districts were calculated. Number of study subjects by each districts were determined proportionally population. A list of study participants were obtained from primary sampling units--policlinics (Primary Health Care Units--PHCUs) in all ten districts of Tbilisi. A fieldwork was arranged for interviewing of selected individuals--Approaching the study subjects in households, Face-to-face interview which included signing of the letter of consent by study subject and administering the questionnaires by an interviewer. In totally 2000 persons were investigated and accordingly 2000 blood samples were tested by ELISA for detection of HCV antibodies. 138 out of 2000 (6.9%) samples were found ELISA positive. 138 ELISA positive samples were tested with more specific Recombinant Immunoblot Assay (RIBA) for confirmation. 134 out of 138 (6.7%) of investigated samples were confirmed by RIBA as positives. In this survey, we found that 134 (6.7%) of the 2,000 surveyed individuals were HCV seropositive. According our study, which was based on very strict epidemiological design, we concluded that prevalence of HCV in General population of Georgia is 6.7%. The part of our investigation was to assess HCV genotypes distribution among general population of Georgia. Based on our results the following distributions of HCV genotypes were found: HCV 1b--59%, HCV 3a--27%, HCV 2a/2c--11%, HCV 1a--3%. Our study found high prevalence of HCV among general population of Georgia. Besides, these surveys found an extensive spread of HCV 1b genotype. The profile of HCV genotypes distribution in general population of Georgia was similar to that of USA and Russia and different compared to Asia, Africa and most of European countries. Unfortunately the HCV genotype 1b is less sensitive to current treatment regimens. The treatment effectiveness in patients with HCV genotype 1b is about 45% in comparison to 80% for non-1 genotypes. PMID- 19124922 TI - Expansion of CD3/CD16/CD56 positive NKT cells in HIV/AIDS: the pilot study. AB - NKT cells are a subset of lymphocytes possessing features of NK cells and T cells; they play a key role in the formation of innate immune response. Upon stimulation, rapid production of large quantities of both T(h1) and T(h2) type cytokines permits them to bridge the innate and adaptive immune responses by activating NK cells, T cells, B cells and dendritic cells. Scientific knowledge has been collecting up to date toward the definition of the role of NKT lymphocytes in HIV/AIDS setting. This direction in HIV/AIDS immunopathogenesis is relatively new and quite concerning. The objective of this study was to investigate CD3+/CD16+/CD56+ NKT cell expansion in HIV/AIDS patients and explore its association with virologic and immunologic markers of HIV infection. Retrospective analysis of 30 HIV infected patients data, taken from the database of the laboratory of clinical immunology at the Infectious Diseases, AIDS and Clinical Immunology Research Center, was conducted. RESULTS: there was slightly increased risk of higher plasma viral load related to lower NKT cell expansion. With regard to immunologic status, borderline significance between expansion of CD3/CD16/CD56 positive NKT cells and lower CD4 positive cell count was shown. However, study did not show strong associations of NKT cell expansion with either virologic or immunologic status, interestingly, all HIV/TB co infected patients where NKT cell positive, which underlines the possible role of TB in CD3+/CD16+/CD56+ NKT cell expansion phenomena in HIV infected individuals. We think these new findings may serve as prerequisite for future, larger scale research in this direction. PMID- 19124923 TI - Fibroscan and FibroTest/FibroMax to assess liver fibrosis/cirrhosis in patients with chronic HBV and HCV infection in Georgia. AB - Although, liver biopsy is the gold standard in assessment of the degree of liver damage, the method has some limitations. For this reason, assessment of liver damage using non-invasive methods is currently an important topic in hepatology. The aim of the study was to evaluate liver fibrosis/cirrhosis using Transient Elastography and FibroTest/FibroMax in patients with chronic HCV and HBV infection in Georgia and to compare Fibroscan and FibroTest/FibroMax results. 252 patients were included in the study, among them 185 with chronic HCV infection and 67 with chronic HBV infection. These patients were investigated at the Georgian-French Joint Hepatology Clinic "Hepa", from December 2007 to November 2008. In patients with chronic HCV or HBV infection Fibroscan and Fibrotest/FibroMax results were correlated in 127 (68.6%) and 45 (67.2%) cases, respectively. Discordance in one degree of fibrosis stage was found in 36 (19.5%) patients with chronic HCV infection and in 14 (20.9%) patients with chronic HBV infection. Discordance in more then one degree of fibrosis stage was found in 22 (11.9%) and 8 (11.9%) cases. In patients with Fibroscan and Fibrotest/FibroMax concordant results liver biopsy might be avoided. Fibroscan and Fibrotest/Max appear to be very valuable methods for detecting early stages of fibrosis among patients with chronic HCV and HBV infection, allowing to avoid the progression of liver damage, as well as end-stage liver disease. These methods are easy to perform and therefore allows regular follow-up of the course of LF. PMID- 19124924 TI - Overview of HIV epidemiological situation in Georgia. AB - Georgia still belongs to low HIV epidemic countries and by December 1st, 2008 there are 1825 HIV/AIDS cases registered at the IDACIRC with estimated number 3500 (estimated prevalence 0.09%). Majority of HIV/AIDS patients are male (75%). Four hundred and sixty one patients are under the Antiretroviral (ARV) treatment, including 23 children. Despite of HIV low prevalence, Georgia is considered to be at risk for an imminent epidemic spread of HIV mainly due to wide spread drug use with high risk practices (needle-sharing) and high rate of STIs. The major route of HIV transmission is associated with IDU. At the moment approximately 60% of all reported HIV cases are due to drug injection. However over the last several years heterosexual route of transmission is gaining importance, and increased from 29.1% to 36.1% for last five years. First significant increase of HIV incidence rate was observed from 1999 to 2000 (2.24 times) and 2003 to 2004. From 2004 stabile, but slight increase of incidence rate is presented. Most HIV positive patients are diagnosed at the age from 25 to 45. The highest HIV prevalence rates are found in Western Georgia, particularly Black Sea coast regions--Megrelia and Adjara (with prevalence of 131.11 and 132.03 among adult HIV cases per 100,000 adult population). Enlarging educational activities, prevention interventions, better financing of HIV programs and improvement of capacity building will help the country to keep HIV epidemic in a low prevalence and give country possibility to achieve "Universal Access to HIV Prevention, Treatment, Care and Support" for 2010 year. PMID- 19124925 TI - The role of imaging studies in CNS infections. AB - Recent advances in nervous system imaging techniques have greatly improved the diagnosis and treatment of central nervous system infections. Nowadays, the most useful imaging modalities are harmless to the patient and widely available. Whereas Analysis of CSF, biopsy, and laboratory analysis remain the gold standard to identify the infectious agent for instance in meningitis, neuroimaging is crucial in clearly depicting inflammatory lesions of brain and spine. Notably, in opportunistic disease neuroimaging has a pivotal role not only in diagnosis but also in monitoring therapeutic response. The present review discusses imaging characteristics of bacterial, viral, fungal and prion infections of the central nervous system. PMID- 19124926 TI - Predicting and optimizing response to cardiac resynchronization therapy beyond QRS duration: expanding role of echocardiography. PMID- 19124927 TI - Coronary artery calcium and you - do you know where you stand? PMID- 19124928 TI - Escalating coronary heart disease and risk factors in South Asians. PMID- 19124930 TI - Myocardial contrast echo: current status and future directions. AB - Contrast echocardiography has been around since the late 1960's. CE began with an M-mode observation by Joyner and a subsequent publication by Gramiak and Shah on enhanced echocardiographic imaging of saline in the aorta. MCE has come a long way from a diagnostic tool to include therapeutics in its armamentarium. It is an ingenious modality for assessment of MPI at the tissue level with special applications in vascular inflammation and drug delivery however, is limited by the technical difficulties with image acquisition and microbubble physics. Refining imaging software and newer advances in microbubble agents continue to revamp contrast echocardiography. PMID- 19124929 TI - Evaluation critique of state of the art dyslipidemia management in general and with a special emphasis on the Indian population. AB - Classically, there have been three well established major cardiovascular risk factors, hypercholesterolemia, hypertension and tobacco abuse. With accumulating clinical evidence, diabetes can now be added as a fourth major risk factor. Much interest in various other risk factors and possible causative factors has been generated, but it should be remembered that of all these, low density lipoproteins (LDL) remains the gold standard for evaluating risk. The common perception is that only caucasians in the western world have significant cardiovascular (CV) risk. However, much clinical information to the contrary has accumulated and now it is realized that many other ethnic groups also have significant CV disease, such as in India, especially in the urban population. Dyslipidemias of specific lipoproteins and their treatment is an important part of understanding and managing CV disease and risk. Various plasma factors such as homocysteine and lipoprotein (a) [(a)] have been considered to have definite associations with CV disease, but any treatment benefit remains in doubt. In addition, inflammatory risk factors are considered to be of significant clinical interest, especially high sensitivity C-Reactive protein (hsCRP). Where do these factors fit into routine clinical practice still awaits clarification. Only two of these inflammatory risk (Lp-factors can be tested commercially on a routine clinical basis and these are hsCRP and Lipoprotein-associated Phospholipase A2 Lp PLA2). Their clinical utillity is not established and acceptance is limited: some third party health coverage organizations refuse to pay for such analyses. In the past, women have been looked upon as not having significant CV disease. More recently, evidence suggests that women may have more CV disease than men, and that physicians may have failed to realize this and act accordingly. The true situation is that women have less CV disease than men prior to menopause and then they slowly catch up. However, some women under age 50 have an especially malignant form of CV disease and in these cases, myocardial infarction mortality is twice that of men. The explanation and management is the subject of much clinical investigation. In both India and the western world, perhaps the most important medical problem is the metabolic syndrome (MS) and this combination of CV risk factors multiplies the significance of each. For the difficult patient not tolerant of or sufficiently responsive to conventional therapy, alternative diets and medications can frequently offer just enough benefit in lowering LDL to allow the patient to attain their target level. Future treatments undoubtedly will involve genetics, but for now, aggressive medication use can favorably modify risk although not eliminate it. PMID- 19124931 TI - Coronary calcium scoring in asymptomatic Indian population: correlation with age, gender and risk factors--a prospective study on 500 subjects. AB - BACKGROUND: The present study was conducted with an objective to assess, quantify and describe a reference value (percentile) of coronary calcium score for the asymptomatic adult Indian population and its correlation with gender, age and risk factors and to review the same with the available literature. METHODS AND RESULTS: Five hundred asymptomatic subjects/volunteers were taken from the general population for this study. Calcium scoring was done based on modified Agatston scoring on a four-row multidetector CT scanner with ECG gating. Findings revealed that with increasing age there was increase in the calcium score. Males had a little higher CACS (not significant) compared to females for any defined age group. There was a significant continuous graded relation between the calcium scores and prevalence of CAC deposits with the number of risk factors. Subjects without any risk factors (n = 340) showed absence of coronary calcium in 85.3 and with the presence of two or more risk factors (n = 72) the absence of coronary calcification was seen in 13.9% The percentile reference values of CACS as obtained from this study (please see table in text) CONCLUSION: This study provides a reference percentile of CACS for Indian population which can be utilized to screen and stratify risk of coronary events in asymptomatic individuals using a non invasive method and thus individuals can be subjected to necessary investigations and risk factor intervention. Limitations in our study - Non availability of lipid profiles, comparison and correlation with angiographic studies and follow up of subjects to evaluate the prospective outcome. PMID- 19124932 TI - Prevalence of ischemic heart disease in a rural population of Bangladesh. AB - Bangladeshi people, as other South Asians, have high susceptibility to ischemic heart disease (IHD) but population-based data are lacking in Bangladesh. We investigated for IHD burden in a rural population of Bangladesh in 2001. Information was sought on use of medications for IHD and electrocardiogram was done in 447 adults (157 men and 290 women) aged 20 years or older (mean 40 years). Prevalence of IHD defined by presence of pathological Q wave on electrocardiogram or current medication for IHD is 3.4% (95% confidence interval, 1.7% to 5.1%). Prevalence in men (4.6%, 1.3% to 7.9%) was almost twice than in women (2.7%, 0.8% to 4.6%). However, this sex-difference should be cautiously interpreted because of small sample size for men. It appears that IHD is an important problem even in this traditional rural population of Bangladesh. PMID- 19124933 TI - Fasting plasma homocysteine levels are increased in young patients with acute myocardial infarction from Western India. AB - BACKGROUND: Fasting hyperhomocysteinemia is positively associated with atherothrombosis and acute myocardial infarction in several prospective and retrospective studies. In India folic acid deficiency is not uncommon, and subclinical folic acid deficiency is known to cause hyperhomocysteinemia without thrombocytopenia. OBJECTIVE: To observe the prevalence of plasma hyperhomocysteinemia in a cohort of patients with acute myocardial infarction after 2 weeks of oral folic acid therapy. MATERIAL AND METHODS: A cohort of 120 consecutive patients with acute myocardial infarction below the age of 40 years was tested for fasting hyperhomocysteinemia 8-10 weeks after AMI. Five hundred age and sex matched unrelated controls and 50 family controls were also studied for two mutations, MTHFR C677T and cystathionine Beta synthase (CBS) T833C mutations Parents of the AMI patients were also tested to see hyperhomocysteinemia in the family. The patients were given two weeks of oral folic acid therapy (folviteR - 5mg once daily) and fasting plasma homocysteine levels were measured again and the pattern of response was noted. Patients who responded partially or not responded at all to oral folic acid therapy received intramuscular injection of 1 mg of cyanoco-balamin ( NeurobionR,) and their level of plasma homocysteine were noted 1 week later. RESULTS: Sixty three out of 120 patients showed hyperhomocysteinemia compared to 9% of the age and sex matched controls. Fifty seven therapy percent of hyperhomocysteinemia patients responded completely to oral folvite. Subsequent vitamin B12 & B6 therapy normalized homocysteine levels in only 2/12 partial responders with less than 50 microg/l of plasma homocysteine levels, but none of the folate non responders and partial responders having plasma homocysteine levels above 50 microgm/l responded to the therapy. CONCLUSION: Hyperhomocysteinemia is common amongst young acute myocardial infarction patients from western India. The major cause of hyperhomocysteinemia in young myocardial infarction cases is folic acid deficiency. However a fair number of patients who did not respond to folic acid, also did not respond to vitamin B12 and B6 parenterally suggesting existence of defect in other pathways of homocysteine metabolism in a subset of our patients. PMID- 19124934 TI - Prevention of no flow/slow reflow phenomenon in primary PCI by Nicorandil. AB - AIM: To investigate the efficacy of Nicorandil in preventing no-flow/slow reflow phenomenon in patients with acute myocardial infarction undergoing primary PCI. MATERIALS AND METHODS: From September 2004 to October 2005, 29 patients underwent a primary percutaneous coronary intervention and stenting with nicor-andil as a protocol drug at a dose of 1 mg/hour - this drug was titrated upwards to maximum tolerated dose, with a 2 mg intracoronary bolus given after balloon inflation during PCI. LAD was the infarct related artery in 62% of cases. 72.4% of them had a TIMI thrombus grade of 5. The corrected TIMI frame count following primary percutaneous intervention in the Nicorandil arm was 19.54 + 8.7. None of the patients had a no flow or slow reflow phenomenon with this protocol. One patient developed a subacute stent thrombosis necessitating a revascularization. At a mean follow up of 251 +/- 96.7% days, MACE was not reported in the other patients. Thirty four patients underwent a primary percutaneous coronary intervention and stenting without nicorandil as an adjuvant drug. Some of these patients were retrospectively assessed. They have been followed up for 285.4 +/- 264.6 days. LAD was the infarct related artery in 61.8% of cases while 79.5% of them had a TIMI thrombus grade of 5. The corrected TIMI frame count in this group was 23.9 +/- 17.5 (p <0.56). MACE was reported in 5 of these patients. The mean TIMI frame count for these 5 patients was 40.5 +/- 29.2. Glycoprotein IIb/IIIa receptor inhibitors were given to all patients in both groups. The choice of the agent used was left to the discretion of the operator. CONCLUSION: Nicorandil prevents no-flow/slow reflow phenomenon in patients undergoing primary PCI for acute myocardial infarction. This is shown by a lower corrected TIMI frame count in the nicorandil arm (p < 0.56). Reduction in the incidence of no-flow/slow reflow phenomenon translates into a lower MACE. The drug is safe and does not require intensive monitoring. It must be started early and electively in patients undergoing a primary PCI as a strategy to prevent no-flow rather than to treat this phenomenon. PMID- 19124935 TI - LV based pacing in patients with heart failure and a narrow QRS--an acute hemodynamic study. AB - BACKGROUND: Biventricular pacing is beneficial in refractory systolic heart failure having QRS duration more than 130 msec by improving regional dysynchrony and decreasing diastolic mitral regurgitation. Current data show significant systolic dysynchrony in symptomatic systolic heart failure patients out of which nearly 40% have a QRS duration of less than 120 msec. Our study aims at assessing acute hemo-dynamic impact of Biventricular (BiV) and compare it with isolated left ventricular (LV) pacing in patients of systolic heart failure and QRS duration < or = 120 msec. METHODS: Seven patients with symptomatic systolic heart failure with LV Ejection fraction (LVEF) < or = 35% (mean 25.7 +/- 11.3%). NYHA functional class more than II and QRS duration < or = 120 msec (mean 92.8 +/- 17.0 msec) were studied at baseline and following BiV and LV pacing with AV delay 100 msec for 5 minutes in random order. Parameters analyzed were heart rate, systolic BP, pulse pressure, LV dimension, LVEF, cardiac output(CO), LV dP/dT, LV and RV isovolumic contraction time and aorto pulmonary flow delay. Duration of QRS complex at baseline and following pacing was noted. 'Responders' were defined as having increase in CO by at least 10% of mean basal cardiac output in study group. RESULTS: BiV pacing resulted in significant improvement in systolic BP (140.71 +/- 21.33 vs 149.29 +/- 19.67 mmHg, p = 0.02), pulse pressure (58.14 +/- 21.14 vs 67.29 +/- 19.57 mmHg, p = 0.01), LVEF (25.71 +/- 11.3 vs 32.86 +/- 4.60%, p = 0.01), CO (3.24 +/- 1.05 vs 3.89 +/- 1.1 l/min, p = 0.02) and LV dP/dT (0.69 +/- 0.22 vs 1.00 +/- 0.23 mmHg/msec, p = 0.001) with a trend towards reduction in LV isovolumic contraction time (115.28 +/- 21.61 vs 99.29 +/- 17.18 msec, p = 0.14) and aorto pulmonary flow delay (25.14 +/- 24.36 vs 12.14 +/- 36.15 msec, p = 0.32). LV pacing resulted in a trend towards improvement in parameters as compared to baseline, systolic BP (140.71 +/- 21.33 vs 146.71 +/- 23.03 mmHg, p = 0.16); pulse pressure (58.14 +/- 21.14 vs 63.29 +/- 26.59 mmHg, p = 0.2); LVEF (25.71 +/- 11.3 vs 33.27 +/- 10.0, p = 0.06); CO (3.24 +/- 1.05 vs 3.27 +/- 0.6 l/min, p = 0.88); LV dP/dT (0.69 +/- 0.22 vs 0.96 +/- 0.39 mmHg/msec, p = 0.16); LV isovolumic contraction time (115.28 +/- 21.61 vs 98.21 +/- 21.34 msec, p = 0.18); aortopulmonary flow delay (25.14 +/- 24.36 vs 5.21 +/- 30.1 msec, p = 0.2). Biventricular and LV pacing resulted in a non-significant increase in duration of paced QRS complexes (105.43 +/- 14.82 msec, p = 0.11 and 108.86 +/- 19.73, p = 0.15 respectively) as compared to 92.86 +/- 17.04 msec at baseline. Three out of 7 patients could be classified as 'responders' to biventricular pacing. CONCLUSION: BiV pacing, and not LV pacing, benefits patients of systolic heart failure (EF < or = 35%) and narrow QRS (< or = 120 msec) on surface ECG. PMID- 19124936 TI - A study of ventricular asynchrony in patients with varying QRS duration and its correlation with left ventricular function. AB - BACKGROUND: Biventricular pacing (cardiac resynchronization therapy) has been shown to improve patients of dilated cardiomyopathy (DCMP), which include largely those with broad QRS complex (LBBB) and a few with normal QRS duration having associated interventricular and/or intraventricular delay. The true prevalence of ventricular asynchrony in a broad subset of patients of DCMP however has not been studied at large and it is unclear if there is association of asynchrony with the LV function. The present study was therefore conducted to assess the prevalence of ventricular asynchrony in a broad set of patients of dilated cardiomyopathy with varying QRS duration and also trying to correlate the same with the LV function. METHOD AND RESULTS: Ninety three consecutive patients of DCMP (EF < 40%) with varying QRS duration and twenty patients with broad QRS complexes (LBBB morphology) with normal LV function (LVEF > 40%) were studied. The patients were then grouped as Group-I, including patients of DCMP with broad QRS (>120 msec, LBBB morphology) (n = 40), Group-II, including patients of DCMP with narrow QRS, (QRS width < or = 120 msec) (n = 53) and Group-III, including patients of LBBB with normal LV function (n = 20). Ventricular asynchrony was diagnosed using standard echocardiography criteria on 2-D, M-mode and pulse wave Doppler including, 1) Interventricular delay:- diagnosed when the difference between aortic (Q-AV) and pulmonary ejection delays (Q-PV) was 340 msec and 2) Intraventricular delay:- diagnosed when posterior left ventricular activation occurred later than onset of diastolic mitral inflow or septal to post wall motion delay of > or = 130 msec. Ventricular asynchrony was present in 97.5% (39/40) in Group-I, 50.9% (27/53) in Group-II and 90% (18/20) in Group-III. Amongst this however interventricular conduction delay was present in 97.5% (39/40) of group-I, 20.7% (11/53) of group-II and 90% )18/20) of group-III, while intraventricular delay was present in 62.5% (25/40) of Group-I, 43.3% (23/53) of Group-II and none of Group-III. Analyzing the contribution of LV dysfunction independent of QRS duration to the occurrence of LV dyssynchrony, patients were further divided in three groups, viz. DCM with LVEF < 20% (group A) (mean LVEF = 16.53 +/- 2.28%), DCM with LVEF > or = 20% (group-B) (mean LVEF = 27.15 +/- 4.29%) and LBBB with normal LV function (group C) (mean LVEF = 53.45 +/- 4.83%). The mean QRS durations were not different between groups A and B (114.3 +/- 32.2 msec vs. 109.1 +/- 13.9 msec, respectively, p = 0.16). However group A had a significantly higher number of patients with ventricular asynchrony compared to group B [37/45 (82.2%) vs. 29/48 (60.4%), p = 0.001)]. CONCLUSION: We conclude that there is a wide prevalence of, but heterogeneity in occurrence of ventricular asynchrony among patients of dilated cardiomyopathy. The asynchrony possibly relates more to left ventricular function in addition rather than to the QRS duration alone. Criteria for biventricular pacing probably need to relate to ventricular asynchrony than to the QRS duration alone. PMID- 19124937 TI - Oral hypoglycemic agents and heart. PMID- 19124938 TI - Acute penetrating atherosclerotic ulcer presenting as paraplegia and renal failure. PMID- 19124939 TI - Non-expansion of carotid stent due to detached stent sheath--an unusual complication of carotid angioplasty treated by surgery. PMID- 19124940 TI - Malignant ventricular arrhythmias due to Aconitum napellus root. PMID- 19124941 TI - All those tachy's are not ventricular. PMID- 19124942 TI - Mobile intracardiac thrombi: clinical scenarios and management strategies. PMID- 19124943 TI - Bivalirudin: a new promising direct antithrombin. AB - The deadliest manifestations of ischemic heart disease are initiated and propagated by intra-coronary thrombin generation. Thrombin is resistant to inactivation by heparin when it is bound to fibrin, fibrin degradation products or subendothelial collagen. Recognition of these limitations has led to development of a new class of antithrombin agents which directly target the active sites on the surface of thrombin molecule and are therefore designated as direct antithrombins. These agents do not need mediation of antithrombin III for their action and are not inhibited by platelet factor 4. This report focuses on bivalirudin, a new agent of promising impact on both interventional as well as non-interventional cardiology. It is a short acting anticoagulant which bivalently and directly inhibits thrombin (coagulation factor II). It binds the active (catalytic) site and the fibrinogen-binding site (exosite I). This provides high affinity and specificity for thrombin. Slow cleavage at the Arg3 Pro4 bond results in recovery of thrombin activity after discontinuation of bivalirudin. Bivalirudin inhibits both protease activated receptor 1 and 4 (PAR 1 and PAR 4) thereby effectively inhibiting acute thrombin mediated platelet aggregation. Clinical efficacy has been assessed and proved in over 20 published patient series focussing on patients with acute coronary syndrome with or without myocardial infarction, patients undergoing percutaneous coronary interventions, patients receiving various adjunctive anti-platelet medications, patients with heparin induced thrombocytopenia or patients undergoing cardiac surgery. In contrast to the well established unfractionated heparin, bivalirudin lacks the risk of heparin induced thrombocytopenia. It shows a tendency to lower bleeding risks without reduction of efficacy when compared with the two-pronged treatment with unfractionated heparin and glycoprotein IIb/IIIa inhibitors. PMID- 19124946 TI - Toward contrast-enhanced microwave-induced thermoacoustic imaging of breast cancer: an experimental study of the effects of microbubbles on simple thermoacoustic targets. AB - Microwave-induced thermoacoustic tomography (MI-TAT) is an imaging technique that exploits dielectric contrast at microwave frequencies while creating images with ultrasound resolution. We propose the use of microbubbles as a dielectric contrast agent for enhancing the sensitivity of MI-TAT for breast cancer detection. As an initial investigation of this concept, we experimentally studied the extent to which the microwave-induced thermoacoustic response of a dielectric target is modified by the presence of air-filled glass microbubbles. We created mixtures of ethylene glycol with varying weight percentages of microbubbles and characterized both their microwave properties (0.5-6 GHz) and thermoacoustic response when irradiated with microwave energy at 3 GHz. Our data show that the microbubbles considerably lowered the relative permittivity, electrical conductivity and thermoacoustic response of the ethylene glycol mixtures. We hypothesize that the interstitial infusion of microbubbles to a tumor site will similarly create a smaller thermoacoustic response compared to the pre-contrast agent response, thereby enhancing sensitivity through the use of differential imaging techniques. PMID- 19124947 TI - Small field measurements with a novel silicon position sensitive diode array. AB - DOSI, a novel dosimeter based on position sensitive detectors for particle physics experiments, was used for relative clinical dosimetry measurements in small radiotherapy fields. The device is capable of dynamic measurements in real time and provides sub-millimetre spatial resolution. The basic beam data for a stereotactic radiotherapy collimator system (BrainLAB) using 6 MV photons were measured and compared with the corresponding data acquired with a small diamond detector and a PinPoint ionization chamber. All measurements showed an excellent agreement between DOSI and the diamond detector. There was an increasing discrepancy between the relative output factors (ROF) measured with DOSI and those measured with the ionization chamber with decreasing field size, specifically for collimators with a diameter smaller than 15 mm. The percentage depth doses (PDD) were in agreement to better than 1% for all depths. The agreement on off-axis ratios (OAR) was better than 3% for all collimators, whereas the agreement on relative output factors (ROF) was at the 1% level. DOSI's fast read-out electronics made it possible for all measurements to be recorded within 45 min including time to change collimators. This should reduce the overall time for commissioning and QA measurements, an important factor especially for busy radiotherapy departments. PMID- 19124948 TI - Probabilistic finite element analysis of radiofrequency liver ablation using the unscented transform. AB - The main limitation of radiofrequency (RF) ablation numerical simulations reported in the literature is their failure to provide statistical results based on the statistical variability of tissue thermal-electrical parameters. This work developed an efficient probabilistic approach to hepatic RF ablation in order to statistically evaluate the effect of four thermal-electrical properties of liver tissue on the uncertainty of the ablation zone dimensions: thermal conductivity, specific heat, blood perfusion and electrical conductivity. A deterministic thermal-electrical finite element model of a monopolar electrode inserted in the liver was coupled with the unscented transform method in order to obtain coagulation zone confidence intervals, probability and cumulative density functions. The coagulation zone volume, diameter and length were 10.96 cm(3), 2.17 cm and 4.08 cm, respectively (P < 0.01). Furthermore, a probabilistic sensitivity analysis showed that perfusion and thermal conductivity account for >95% of the variability in coagulation zone volume, diameter and length. PMID- 19124949 TI - Analytically derived weighting factors for transmission tomography cone beam projections. AB - Weighting factors, which define the contributions of individual voxels of a 3D object to individual projection elements (pixels) on the detector, are the basic elements required in iterative tomographic reconstructions from transmission projections. Exact or as accurate as possible values for weighting factors are required in high-resolution reconstructions. Geometric complexity of the problem, however, makes it difficult to obtain exact weighting factor values. In this work, we derive an analytical expression for the weighting factors in cone beam projection geometry. The resulting formula is validated and applied to reconstruction from mega and kilovoltage x-ray cone beam projections. The reconstruction speed and accuracy are significantly improved by using the weighting factor values. PMID- 19124950 TI - Significance of background optical properties, time-resolved information and optode arrangement in diffuse optical imaging of term neonates. AB - The significance of accurate knowledge of background optical properties and time resolved information in reconstructing images of hemodynamic changes in the neonatal brain from diffuse optical imaging data was studied using Monte Carlo (MC) simulation. A segmented anatomical magnetic resonance (MR) image and literature-derived optical properties for each tissue type were used to create a voxel-based anatomical model. Small absorbing perturbations were introduced into the anatomical model to simulate localized hemodynamic responses related to brain activation. Perturbation MC (pMC) was used as the primary method of image reconstruction. For comparison, reconstructions were also performed using the finite element method (FEM) to solve the diffusion approximation (DA) to the radiative transfer equation (RTE). The effect of optode layout was investigated using three different grids. Of the factors studied, the density of the optode grid was found to have the greatest effect on image quality. The use of time resolved information significantly improved the spatial accuracy with all optode grids. Adequate knowledge and modeling of the optical properties of the background was found to significantly improve the spatial accuracy of the reconstructed images and make the recovery of contrast of absorption changes more consistent over simplified modeling. Localization accuracy of small perturbations was found to be 2-3 mm with accurate a priori knowledge of the background optical properties, when a grid with high optode density (>1 optode cm(-2)) was used. PMID- 19124951 TI - The influence of anatomical and physiological parameters on the interference voltage at the input of unipolar cardiac pacemakers in low frequency electric fields. AB - The problem of electromagnetic interference of electronic implants such as cardiac pacemakers has been well known for many years. An increasing number of field sources in everyday life and occupational environment leads unavoidably to an increased risk for patients with electronic implants. However, no obligatory national or international safety regulations exist for the protection of this patient group. The aim of this study is to find out the anatomical and physiological worst-case conditions for patients with an implanted pacemaker adjusted to unipolar sensing in external time-varying electric fields. The results of this study with 15 volunteers show that, in electric fields, variation of the interference voltage at the input of a cardiac pacemaker adds up to 200% only because of individual factors. These factors should be considered in human studies and in the setting of safety regulations. PMID- 19124952 TI - Risk of second primary cancer following prostate cancer radiotherapy: DVH analysis using the competitive risk model. AB - This study aimed to estimate the risk of developing second primary cancer (SPC) corresponding to various radiation treatment techniques for prostate cancer. Estimation of SPC was done by analysing differential dose-volume histograms (DDVH) of normal tissues such as rectum, bladder and urethra with the competitive risk model. Differential DVHs were obtained from treatment planning systems for external beam radiotherapy (EBRT), low-dose-rate (LDR) and high-dose-rate (HDR) brachytherapy techniques. The average risk of developing SPC was no greater than 0.6% for all treatment techniques but was lower with either LDR or HDR brachytherapy alone compared with any EBRT technique. For LDR and HDR brachytherapy alone, the risk of SPC for the rectum was 2.0 x 10(-4)% and 8.3 x 10(-5)% respectively compared with 0.2% for EBRT using five-field 3D-CRT to a total dose of 74 Gy. Overall, the risk of developing SPC for urethra following all radiation treatment techniques was very low compared with the rectum and bladder. Treatment plans which deliver equivalent doses of around 3-5 Gy to normal tissues were associated with higher risks of development of SPC. PMID- 19124954 TI - A robust Hough transform algorithm for determining the radiation centers of circular and rectangular fields with subpixel accuracy. AB - Uncertainty in localizing the radiation field center is among the major components that contribute to the overall positional error and thus must be minimized. In this study, we developed a Hough transform (HT)-based computer algorithm to localize the radiation center of a circular or rectangular field with subpixel accuracy. We found that the HT method detected the centers of the test circular fields with an absolute error of 0.037 +/- 0.019 pixels. On a typical electronic portal imager with 0.5 mm image resolution, this mean detection error was translated to 0.02 mm, which was much finer than the image resolution. It is worth noting that the subpixel accuracy described here does not include experimental uncertainties such as linac mechanical instability or room laser inaccuracy. The HT method was more accurate and more robust to image noise and artifacts than the traditional center-of-mass method. Application of the HT method in Winston-Lutz tests was demonstrated to measure the ball-radiation center alignment with subpixel accuracy. Finally, the method was applied to quantitative evaluation of the radiation center wobble during collimator rotation. PMID- 19124953 TI - Monte Carlo simulations to assess the effects of tube current modulation on breast dose for multidetector CT. AB - Tube current modulation was designed to reduce radiation dose in CT imaging while maintaining overall image quality. This study aims to develop a method for evaluating the effects of tube current modulation (TCM) on organ dose in CT exams of actual patient anatomy. This method was validated by simulating a TCM and a fixed tube current chest CT exam on 30 voxelized patient models and estimating the radiation dose to each patient's glandular breast tissue. This new method for estimating organ dose was compared with other conventional estimates of dose reduction. Thirty detailed voxelized models of patient anatomy were created based on image data from female patients who had previously undergone clinically indicated CT scans including the chest area. As an indicator of patient size, the perimeter of the patient was measured on the image containing at least one nipple using a semi-automated technique. The breasts were contoured on each image set by a radiologist and glandular tissue was semi-automatically segmented from this region. Previously validated Monte Carlo models of two multidetector CT scanners were used, taking into account details about the source spectra, filtration, collimation and geometry of the scanner. TCM data were obtained from each patient's clinical scan and factored into the model to simulate the effects of TCM. For each patient model, two exams were simulated: a fixed tube current chest CT and a tube current modulated chest CT. X-ray photons were transported through the anatomy of the voxelized patient models, and radiation dose was tallied in the glandular breast tissue. The resulting doses from the tube current modulated simulations were compared to the results obtained from simulations performed using a fixed mA value. The average radiation dose to the glandular breast tissue from a fixed tube current scan across all patient models was 19 mGy. The average reduction in breast dose using the tube current modulated scan was 17%. Results were size dependent with smaller patients getting better dose reduction (up to 64% reduction) and larger patients getting a smaller reduction, and in some cases the dose actually increased when using tube current modulation (up to 41% increase). The results indicate that radiation dose to glandular breast tissue generally decreases with the use of tube current modulated CT acquisition, but that patient size (and in some cases patient positioning) may affect dose reduction. PMID- 19124955 TI - A novel unidirectional intensity map segmentation method for step-and-shoot IMRT delivery with segment shape control. AB - In intensity modulated radiation therapy (IMRT), intensity maps are computed from prescribed doses to target volumes, adding dose restrictions to the surrounding tissues. Those intensity (fluence) maps are discretized into matrices of natural numbers and translated to sequences of multileaf collimator (MLC) leaf movements, which will finally deliver the computed x-ray intensities. A unidirectional leaf sequencing algorithm that controls the shape of the segments and reduces leaf motion time for step-and-shoot dose delivery is presented. The problem of constructing segments in controlling its shape was solved by synchronizing right leaves motion. This is done without increasing the number of segments, or the total number of monitor units, and taking into account the unidirectional leaf motion and the interleaf collision constraints. The method was tested using random matrices and a clinical case planned with the PCRT 3D(R) planning system. Compared to other unidirectional leaf sequencing methods, the proposed algorithm performs very similarly. But, in addition, the segment shape control produces segments with smoother outlines and more compact shapes, which may help to reduce MLC-specific effects when delivering the planned fluence map. Finally, as a result of imposing unidirectionality, this algorithm can be extended for sliding window segment generation. PMID- 19124957 TI - [Acute infectious encephalitis and pathogens coming from animals]. AB - Despite better knowledge of pathophysiology and a wider use of new molecular technologies for the diagnosis, the etiological diagnosis of acute encephalitis is not established in most cases. Incidence, prognosis, rate of this disease and severity of sequelae remain unknown. In France, according to the published data, the incidence of encephalitis is estimated to be 1.9 cases per 100 000 inhabitants in average among non-HIV patients. The etiological diagnosis is established in less than 30% of cases. The more frequent diagnosis is herpetic encephalitis in adults and encephalitis caused by Varicella zoster virus in children younger than 16 years. Despite a difficult diagnosis and the lack of specific treatment for most of these infections, the etiological diagnosis should always be deeply explored to precise the individual prognosis, to allow better management of antibiotic therapy, and to improve epidemiological knowledge. We present the recommendations established by the French Society for Infectious Diseases. First designed to suit the French epidemiology, they take in count the possible exposure of patients to different epidemiological patterns. Three levels of etiological tests are proposed, from the most common infections and those, which required an immediate treatment, to the rarest ones. PMID- 19124956 TI - Exact and approximate Fourier rebinning of PET data from time-of-flight to non time-of-flight. AB - The image reconstruction problem for fully 3D TOF PET is challenging because of the large data sizes involved. One approach to this problem is to first rebin the data into one of the following lower dimensional formats: 2D TOF, 3D non TOF or 2D non TOF. Here we present a unified framework based on a generalized projection slice theorem for TOF data that can be used to compute each of these mappings. We use this framework to develop approaches for rebinning into non TOF formats without significant loss of information. We first derive the exact mappings and then describe approximations which address the missing data problem for oblique sinograms. We evaluate the performance of approximate rebinning using Monte Carlo simulations. Our results show that rebinning into non TOF sinograms retains significant SNR advantages over sinograms collected without TOF information. PMID- 19124958 TI - Selected environmental risk factors and congenital heart defects. AB - The aim of the article is to review the published scientific literature and epidemiological studies about the effect of selected environmental risk factors on congenital heart defects in infants. According to recent reports, the prevalence of congenital heart defects is around 1% of live births. Congenital heart malformations are the leading cause of infant mortality. Unfortunately, the majority of the causes of heart defects remain unknown. These malformations are caused by interaction of genetic and environmental factors. The article reviews selected environmental risk factors: maternal illnesses and conditions associated with metabolic disorder (maternal diabetes, obesity, phenylketonuria), maternal lifestyle factors (alcohol use, smoking), which may increase the risk of congenital heart defects. PMID- 19124959 TI - [C-reactive protein levels in patients with chronic obstructive pulmonary disease and asthma]. AB - Chronic obstructive pulmonary disease (COPD) and asthma are defined as chronic inflammatory airway diseases. There is increasing evidence that systemic inflammation may be involved in their pathogenesis too. We aimed to investigate the C-reactive protein levels in plasma of patients with COPD, asthma and control subjects and to evaluate associations of C-reactive protein levels with pulmonary function and smoking history. MATERIAL AND METHODS: We investigated 87 persons: 41 with COPD, 30 with asthma, and 16 controls. Clinical evaluation, pulmonary function tests, C-reactive protein concentration measurement, body mass index and smoking history evaluation were performed. RESULTS: We determined significantly higher C-reactive protein concentrations in COPD patients compared with asthma patients and controls (8.37+/-1.14 vs 3.14+/-0.67 and 2.39+/-0.59 mg/L, respectively; P<0.001). C-reactive protein concentrations in smokers and ex smokers with COPD were significantly higher than in COPD non-smokers (8.38+/-1.52 and 10.4+/-2.22 vs 4.10+/-0.86 mg/L, respectively; P<0.05). In COPD patients, C reactive protein level correlated with FEV(1) (R=-0.463, P=0.002), FEV(1)/FVC (R= 0.449, P=0.003), and pack-years (R=0.572, P=0.001). There was no correlation between C-reactive protein level and analyzed parameters in asthmatics and control group. CONCLUSIONS: Our data support the hypothesis that systemic inflammation plays a role in the pathogenesis of COPD, and cigarette smoking might influence this inflammation. PMID- 19124960 TI - [Return to work after coronary artery bypass surgery]. AB - The aim of this study was to assess the possible reasons for not returning to work after coronary artery bypass surgery. A total of 134 patients (aged 65 years and younger) who underwent coronary bypass surgery in 2003 were examined. The analysis was performed in three groups of the patients: Group I, patients who were employed before surgery and returned to work after it (n=51); Group II, patients who were employed before surgery but did not return to work after surgery (n=55); and Group III, patients who were unemployed before and remained unemployed after surgery due to health problems (n=28). Number of injured coronary arteries, the extent of operation, postoperative complications, risk factors for ischemic heart disease, clinical status of patients (angina pain and heart failure), physical tolerance, and return to work within one year after coronary bypass surgery were analyzed. It was found that 48.1% of patients who were employed before surgery returned to work after myocardial revascularization. About 30% of patients experienced recurrent symptoms of angina after 12 months. Logistic regression analysis revealed that return to work was significantly influenced by female gender, physical pattern of work, age, and severity of heart failure. PMID- 19124961 TI - Cardiomyocyte remodeling in ischemic heart disease. AB - OBJECTIVE: The aim of the study was to detect changes in left ventricular cardiomyocyte size and shape in response to chronic ischemia and loss of cardiac tissue (myocardial infarction) during the course of ischemic heart disease (IHD). MATERIAL AND METHODS: Left ventricular cardiomyocyte dimensions (diameter and length) were estimated histomorphometrically, and their cross-sectional area and volume were assessed in 85 males who died suddenly out of hospital (within 6 hours of the onset of the terminal event) due to the acute first (preinfarction IHD group, n=53, aged 48.6+/-2.9 years) or repeated (postinfarction IHD group, n=32, aged 51.7+/-2.9 years) IHD attack, and had no other causes for the increased heart load. Twenty-nine males of similar age (mean age, 46.0+/-3.1 years) who succumbed to external causes served as controls. RESULTS: We have found cardiomyocyte hypertrophy in the preinfarction IHD group already. The cardiomyocyte volume was increased by 32.0% in comparison with the same index in the control group, and cross-sectional area and length--by 17.2 and 12.5%, respectively. In postinfarction IHD group, all studied cardiomyocyte parameters did not differ significantly from the analogous indices in the preinfarction IHD group (P>0.05). Cardiomyocyte hypertrophy was related to the increase in left ventricular cardiomyocyte parameters. CONCLUSIONS: Left ventricular cardiomyocyte hypertrophy occurs before the first myocardial infarction. In postinfarction myocardium, cardiomyocyte dimensions do not differ significantly at least prior to the appearance of congestive heart failure syndrome. PMID- 19124962 TI - Umbilical hernia: factors indicative of recurrence. AB - Umbilical hernia has gained little attention from surgeons in comparison with other types of abdominal wall hernias (inguinal, postoperative); however, the primary suture for umbilical hernia is associated with a recurrence rate of 19 54%. The aim of this study was to analyze the results of the umbilical hernia repair and to assess the independent risk factors influencing umbilical hernia recurrence. MATERIALS AND METHODS: A retrospective analysis of patients who underwent surgery for umbilical hernia in the Hospital of Kaunas University of Medicine in 2001-2006 was performed. Age, sex, hospital stay, hernia size, patient's body mass index, and postoperative complications were analyzed. Postoperative evaluation included pain and discomfort in the abdomen and hernia recurrence rate. The questionnaire, which involved all these previously mentioned topics, was sent to all patients by mail. Hernia recurrence was diagnosed during the patients' visit to a surgeon. Two surgical methods were used to repair umbilical hernia: open suture repair technique (keel technique) and open mesh repair technique (onlay technique). Every operation was chosen individually by a surgeon. RESULTS: Ninety-seven patients (31 males and 66 females) with umbilical hernia were examined. The mean age of the patients was 57.1+/-15.4 years, hernia anamnesis - 7.6+/-8.6 years, hospital stay - 5.38+/-3.8 days. Ninety-two patients (94.8%) were operated on using open suture repair technique and 5 (5.2%) patients - open mesh repair technique. Only 7% of patients whose BMI was >30 kg/m(2) and hernia size >2 cm and 4.3% of patients whose BMI was < 30 kg/m(2) and hernia size < 2 cm were operated on using onlay technique (P>0.05). The rate of postoperative complications was 5.2%. Sixty-seven patients (69%) answered the questionnaire. The complete patient's recovery time after surgery was 2.4+/-3.4 months. Fourteen patients (20.9%) complained of pain or discomfort in the abdomen, and 7 patients (10.4%) had ligature fistula after the surgery. Forty-five patients (67.2%) did not have any complaints after surgery. The recurrence rate after umbilical hernia repair was 8.9%. The recurrence rate was higher when hernia size was >2 cm (9% for <2 cm vs 10.5% for >2 cm) and patient's BMI was >30 kg/m(2) (8.6% for < 30 vs 10.7% for >30). There were 5 recurrence cases after open suture repair and one case after onlay technique. Fifty-six patients (83.6%) assessed their general condition after surgery as good, 9 patients (13.4%) as satisfactory, and only 2 patients (3%) as poor. CONCLUSIONS: We did not find any significant independent risk factors for umbilical hernia recurrence. However, based on reviewed literature, higher patient's body mass index and hernia size of >2 cm could be the risk factors for umbilical hernia recurrence. PMID- 19124963 TI - The evaluation of the rehabilitation effects on cognitive dysfunction and changes in psychomotor reactions in stroke patients. AB - Stroke patients often experience cognitive dysfunctions. One of the parameters assessing cognitive function is the reaction time as it reflects the speed of information processing. The aim of the study was to assess cognitive and psychomotor dysfunctions and the effectiveness of rehabilitation in stroke patients. The subjects of the study were 30 stroke patients who underwent rehabilitation at the Department of Neurorehabilitation. The mean age of patients was 65.33+/-13.2 years. During the study, the patients' cognitive functions, the handgrip strength, reaction time, and frequency of movements were assessed. There was a substantial improvement in patients' cognitive function after rehabilitation. Assessing the results obtained by Mini Mental State Examination, the change was 6.4+/-2.3 points, and assessing by Neurobehavioral Cognitive Status Examination (Cognistat), the change was 13.3+/-10 points (P<0.05). Before the early stage of rehabilitation, Cognistat results showed that the majority of the patients experienced memory loss, diminished constructive abilities, and disorientation. After occupational therapy, there was a statistically significant improvement in all cognitive function domains. At the end of the inpatient rehabilitation period, there was a significant change in patients' reaction time and movement frequency. At the end of early rehabilitation stage, the estimated reaction time in patients with stroke was compared with reaction time in healthy elderly people of the same age. There was no significant difference between these groups; consequently, we concluded that after rehabilitation, improvement of function was achieved. The results of this study showed that at the beginning of early rehabilitation period and after rehabilitation applied, there was a significant moderate correlation between mental state and reaction time in stroke patients. PMID- 19124964 TI - Pulmonary edema and hemorrhage as complications of acute airway obstruction following anesthesia. AB - Airway obstruction is a quite common complication while its conditioned pulmonary edema--rare. Causes associated with anesthesia are various. Forced inspiratory efforts against an obstructed upper airway generate peak negative intrathoracic pressure. This may cause pulmonary edema and in some cases pulmonary hemorrhage. Last-mentioned is extremely rare. Pulmonary edema may arise soon after airway obstruction as well as later, after some hours. Damage of bronchi is found seldom during bronchoscopy in case of pulmonary hemorrhage, while more often alveolar damage is observed due to alveolar membrane damage. Hemorrhage is conditioned by hydrostatic pressure level, level of hypoxia, damage to bronchi or alveoli (disruption of alveolar membrane). Early diagnosis of negative-pressure pulmonary edema or pulmonary hemorrhage is very important, because this affects postoperative morbidity and mortality of the patients. Two cases of pulmonary edema and hemorrhage after upper airway obstruction as well as literature overview are presented in this article. Pulmonary hemorrhage developed during anesthesia with ketamine, conditioned by increment of hydrostatic pressure, hypoxia, and effects of ketamine on hemodynamics. PMID- 19124965 TI - [Socialization of athletes with disabilities in adapted physical activity]. AB - The aim of this study was to determine the impact of the impairment of the body functions on the participation of people with disabilities in sports activities. MATERIAL AND METHODS: The study was carried out in the institutions mostly attended by people with severe physical disabilities. The participants took a modified Kenny test and answered the questions of a questionnaire. The study sample consisted of 35 persons with severe physical disabilities. RESULTS: The findings of this study showed that people with disabilities who were not engaged in sports were of much worse opinion about their health condition (P=0.02) and they needed more help from family or friends (P=0.035) compared to the disabled who were not engaged in sports, but in the group of people with disabilities who were engaged in sports, the correlations of those indicators were statistically significant (r=0.59 and r=0.68, respectively). The main motivation of sports participation of people with disabilities (about 80%) was the need for communication and gaining independence. Health improvement was mentioned by less than half of people with disabilities (about 41%) engaged in sports. CONCLUSIONS: The syndrome of movement function impairment, duration of impairment, marital status, the age of persons with disabilities, and objectively determined impairment of biosocial self-service functions did not impact sports participation of people with disabilities. However, subjective sensation of pain and the need of medical aid, which did not match the objective functional impairments, could be the obstacle for people with disabilities to participate in sports. PMID- 19124966 TI - [The comparison of patients' and nurses' attitudes to health education and nurses' participation in this process]. AB - The aim of the study is to evaluate and to compare patients' and nurses' opinion about information given to patients by nurses. MATERIALS AND METHODS: The study was performed during April-May 2007 in two regions of Lithuania. Eight hospitals were randomly selected for the study. The study included all patients and nurses who were in therapeutic and surgery departments on the day the survey was carried out. A total of 1030 questionnaires were distributed among patients (the response rate was 85.0%), and 436 questionnaires--among nurses (the response rate was 89.0%). RESULTS: In most cases, patients about their disease are informed by a doctor; this was pointed out by 90.6% of nurses and 90.4% of patients. More than half of patients (65.9%) noted that they knew sufficiently enough about their disease, and 26.4% of nurses agreed. Nurses' and patients' opinion on the adequacy of information provided to patients by nurses differed: more patients than nurses indicated that they got enough information on their disease, health status, duration of intended operation and treatment, but nurses noted that they gave more information on nursing procedures, preparation for forthcoming tests, use of administered drugs, preparation for operation, and healthy lifestyle than was recognized by patients. The patients who were treated in the surgery departments received less information about preparation for forthcoming tests and healthy lifestyle. Patients up to 45 years, city residents, those with higher education, and females were more interested in the above-mentioned information. Nurses who had higher education were more self-critical and more often recognized that they did not provide enough information for patients. CONCLUSIONS: More than half of patients reported that they knew enough about their disease; their awareness depended on their age, sex, place of residence, and education. Physicians were indicated to be the main source of health information in most cases. Nurses more often than patients noted that information provided by them was adequate. The patients who were treated in the surgical departments received less information about healthy lifestyle. PMID- 19124967 TI - [Lithuanian pharmacists in Russia at the beginning of the 20th century: their practice and national patriotic activity]. AB - An important role in the formation of modern Lithuanian society was played by pharmacists who at the beginning of the 20th century were one of the most numerous parts of Lithuanian intelligentsia. They chose a job in a pharmacy not as a mission of life but due to political, social, and economic reasons. The majority of pharmacists were children of peasants who had refused to obey their parents and study in the seminary of priests. Those people who had been ousted from gymnasiums because of an anticzarist activity or those who had not finished school due to the lack of money also became pharmacists. Young men who had chosen a way of self-support left to the biggest cities of Russia and started the practice of an apprentice in a pharmacy. Later, they took examinations to become an assistant of a pharmacist, and after two years of studies at university, they took examinations of a pharmacist at last. Having got a diploma, they usually did not return to their motherland because there was a large network of pharmacies; thus, business conditions were harder, of course. They established pharmacies in various provinces of Russia most often, and it is supposed that only 10% of Lithuanian pharmacists worked in their native country. Living and working in a Russian environment, however, they enshrined national patriotic ideas, were active participants in social activities, published a Lithuanian professional newspaper "Farmaceutu reikalai" (translation, "Matters of pharmacists"), and attempted to unite all Lithuanians living in czarist Russia to struggle against denationalization, to encourage people to return to their motherland, and to work for its good. This article deals with the path for a career of Lithuanian pharmacists in czarist Russia and their national patriotic activity. PMID- 19124968 TI - [Indications for platelet transfusion]. AB - Relevance of platelet transfusion is related to an increasing number of indications due to conditions with an increased risk for hemorrhage and a lack of standard protocols both for treatment and prophylaxis of thrombocytopenia, in the presence of high costs of this procedure. The appropriate use of platelet transfusion is associated with reduction of thrombocytopenia, which actually in a critically ill patient is associated with increased length of stay both in intensive care unit and hospital. In 2003, the Professional Consensus was established to define the main recommendations for platelet transfusion. Not only the number of platelets but also the presence of critical conditions (disorders of thermoregulation, trauma, head brain damage, etc.) increasing the risk for bleeding is relevant. It is recommended that the platelet count in one dose of platelets transfused would be not lower than 55 x 10(9)/L in order to increase the platelet count by 5-10 x 10(9)/L. The effectiveness of platelet transfusion is evaluated 10-60 min, 18 hours, and 24 hours after transfusion. Advantages of platelet transfusion of small and large doses are presented. PMID- 19124971 TI - Cdk1-FOXO1: a mitotic signal takes center stage in post-mitotic neurons. AB - An emerging theme in molecular neurobiology is the discovery of post-mitotic functions for proteins classically associated with mitotic transition in cycling cells. Although neurons have departed the cell cycle, they surprisingly express molecules in the cell cycle apparatus throughout development. The major mitotic cyclin-dependent kinase Cdk1 plays a critical role during the period of naturally occurring neuronal death in the nervous system and has been suggested to contribute to the pathogenesis of neurodegenerative diseases. However, the mechanisms by which Cdk1 promotes neuronal apoptosis are incompletely understood. A recent report by Yuan et al., (2008) has identified a direct relationship between this mitotic kinase and forkhead transcription factor FOXO1, a protein previously implicated in cell death, DNA damage repair and tumor suppression. Here we will discuss the key findings of this report and consider the implications of this mechanism to the regulation of other signal transduction pathways in brain development and diseases. PMID- 19124975 TI - What is the Mechanism of No Injury (MONI)? PMID- 19124976 TI - The relationship of hamstrings and quadriceps strength to anterior cruciate ligament injury in female athletes. AB - OBJECTIVE: To determine the association of quadriceps and hamstrings strength to anterior cruciate ligament (ACL) injury risk in female athletes. The primary hypothesis was that there would be decreased knee flexor and increased knee extensor strength in female athletes who went on to ACL injured status (FACL) compared to uninjured female (FC) and male (MC) control subjects. STUDY DESIGN: Matched case control. SETTING: Institutional Biomechanics Laboratory. PARTICIPANTS: Prospectively measured FACL (n = 22) female athletes who subsequently suffered confirmed noncontact ACL ruptures (16 during soccer and 6 during basketball play) were matched (1:4 ratio) to female controls (FC; n = 88) using limb (dominant or nondominant), pubertal status, sport, and nearest height and mass. In addition, male controls (MC) were matched (1:1 ratio) to FACL to serve as a secondary comparative control. ASSESSMENT OF RISK FACTORS: Isokinetic (concentric) knee extension/flexion strength (300 degrees/s). RESULTS: FACL subjects had decreased hamstrings strength compared to MC (15%; 95% CI, 1 to 27%; P = 0.04). FC were not different from MC in hamstrings strength. Conversely, FACL subjects did not differ compared to the MC in quadriceps strength, and the FC demonstrated decreased quadriceps strength relative to MC (10%; 95% CI, 3 to 18%; P = 0.01). CONCLUSIONS: The results of this investigation indicate that female athletes who suffered ACL injury subsequent to strength testing had a combination of decreased hamstrings strength but not quadriceps strength compared to males. In direct contrast, female athletes who did not go on to ACL injury had decreased quadriceps strength and similar hamstrings strength compared to matched male athletes. PMID- 19124977 TI - The Thessaly test for detection of meniscal tears: validation of a new physical examination technique for primary care medicine. AB - OBJECTIVE: To assess the validity of a new clinical test (Thessaly) as a means of detecting meniscal tears of the knee by comparing arthroscopic findings to a clinical examination finding. DESIGN: Retrospective cohort study. SETTING: All preoperative examinations were performed in the Department of Orthopedic Surgery, a secondary care center, Dwight David Eisenhower Army Medical Center, Fort Gordon, Georgia. PARTICIPANTS: 116 consecutive patients undergoing knee arthroscopy for suspected meniscal pathology. INTERVENTION: The Thessaly test was performed during the preoperative examination as previously described. The clinician supports the patient by holding their outstretched hands while the patient stands flatfooted. The patient then rotates their knee and body, internally and externally, three times, keeping the knee flexed at 20 degrees. Patients with suspected meniscal tears will experience joint-line discomfort. MAIN OUTCOME MEASURES: The Thessaly test performed at 20 degrees of flexion and arthroscopic surgical diagnosis. RESULTS: Of the 66 patients with a positive Thessaly test, 65 had an arthroscopically verified meniscal tear. The Thessaly test revealed a sensitivity of 90.3%, specificity of 97.7%, positive predictive value of 98.5%, negative predictive value of 86.0%, likelihood ratio for a positive test of 39.3, likelihood ratio for a negative test of 0.09, and diagnostic accuracy of 88.8%. The resulting kappa coefficient revealed a statistically significant level of agreement (P < 0.001) for the surgical diagnosis and the Thessaly test. CONCLUSIONS: The Thessaly test is a valid and reproducible physical examination technique for predicting meniscal tears. The Thessaly test shows promise as an easily performed maneuver that may have better diagnostic accuracy than traditional tests. However, this study was performed at a referral center; therefore, the diagnostic relevance cannot be appropriately applied to a more generalized population. PMID- 19124978 TI - Differential emotional responses of varsity athletes to concussion and musculoskeletal injuries. AB - OBJECTIVE: To determine if athletes with concussion and those with minor musculoskeletal injuries experienced differential emotional response to injury. DESIGN: A prospective longitudinal cohort study. SETTING: University of Toronto, Ontario, Canada. PARTICIPANTS: Thirty-four injured athletes from Canadian Interuniversity Sport (CIS) and 19 healthy, physically active undergraduate students participated in the study. INTERVENTION: All participants completed the Profile of Mood States (POMS; short version) on 3 nonconsecutive days during a 2 week period after a baseline test. MAIN OUTCOME MEASURES: Emotional responses were assessed using the POMS. The 7 main outcome measures assessed by POMS were tension, depression, anger, vigor, fatigue, confusion, and total mood disturbance. RESULTS: After injury, concussion produced an emotional profile characterized by significantly elevated fatigue and decreased vigor. In contrast, athletes with musculoskeletal injuries displayed a significant increase in anger that resolved to a pre-injury level within 2 weeks. CONCLUSIONS: The results revealed that both injured groups experienced emotional disturbance after injury. More importantly, the findings strongly suggest that the emotional reaction after concussion is different from that of musculoskeletal injury. Therefore, we concluded that assessing emotional reactions to concussion is particularly important and recommend that sports medicine professionals assess and monitor emotional functioning as well as somatic complaints and neurocognitive changes during recovery. PMID- 19124979 TI - Injury rates and risk factors in competitive professional boxing. AB - OBJECTIVE: To determine injury rates and risk factors for injury in a cohort of professional boxers. DESIGN: Retrospective cohort design reporting on data collected for a fight statistics database maintained by the Professional Boxing and Combat Sports Board of Victoria, Australia. Data were extracted for the years January 1997 through June 2005. SETTING: Victoria, Australia. PARTICIPANTS: 545 professional boxers (age, 18 to 43 years) who participated in a total of 907 fights over the study period. ASSESSMENT OF RISK FACTORS: Independent variables under investigation included age, gender, weight, bout exposure, and location of the bout (within or outside of the State of Victoria). MAIN OUTCOME MEASURES: Physician-reported acute boxing injuries occurring during bouts of any region or nature. RESULTS: 214 injuries were sustained over the 8.5 years, corresponding to an injury rate of 23.6 per 100 professional fights. The majority of these injuries were lacerations to the head and face. An increasing age and an increasing number of fights were both significant predictors of injury. CONCLUSIONS: Injury reduction strategies for professional boxing might include restrictions of eligibility to fight based on age and boxing bout exposure. Future research using prospective cohort designs and standardized injury definitions are needed to confirm these results. Greater mechanistic detail and more complete data entry are necessary to ensure that optimal injury prevention strategies can be developed and implemented. Upon confirmation of the results of this study, the Professional Boxing and Combat Sports Board of Victoria may consider different criteria upon which to sanction a fight. PMID- 19124980 TI - Sports injuries surveillance during the 2007 IAAF World Athletics Championships. AB - OBJECTIVE: The aim of this study was to analyze all sports injuries incurred in competitions and/or training during the 2007 World Athletics Championships and to prove the feasibility of the injury surveillance system developed for the 2008 Olympic Games for individual sports. DESIGN: Prospective recording of injuries. SETTING: 11 IAAF World Championships in Athletics 2007 in Osaka, Japan. PARTICIPANTS: All national team physicians and physiotherapists; Local Organising Committee (LOC) physicians working in the Medical Centres at the stadium and warm up area. MAIN OUTCOME MEASURES: Frequency, characteristics, and incidence of injuries. RESULTS: 192 injuries were reported, resulting in an incidence of 97 injuries per 1000 registered athletes. More than half of the injuries (56%) were expected to prevent the athlete from participating in competition or training. Eighty percent affected the lower extremity; the most common diagnosis was thigh strain (16%). In most cases, the injury was caused by overuse (44%). A quarter of the injuries were incurred during training and 137 (71%) in competition. On average, 72.4 injuries per 1000 competing athletes were incurred in competitions. The incidence of injury varied substantially among the disciplines. The risk of a time-loss injury was highest in heptathlon, women's 10,000 m, women's 3000 m steeplechase, decathlon, and men's marathon. CONCLUSION: The injury surveillance system proved feasible for individual sports. Risk of injury varied among the disciplines, with highest risk in combined disciplines, steeplechase, and long distance runs. Preventive interventions should mainly focus on overuse injuries and adequate rehabilitation of previous injuries. PMID- 19124981 TI - Medication use by athletes at the Athens 2004 Summer Olympic Games. AB - OBJECTIVE: To gather data and examine the use by elite Olympic athletes of food supplements and pharmaceutical preparations in total and per sport, country, and gender. DESIGN: Survey study. SETTING: Athens 2004 Olympic Games (OG). PARTICIPANTS: Data from 2 sources were collected: athletes' declaration of medications/supplements intake recorded on the Doping Control Official Record during sample collection for doping control, and athletes' application forms for granting of a therapeutic use exemption (TUE) and through the abbreviated TUE process (aTUE). MAIN OUTCOME MEASURES: Classification of declared food supplements according to the active ingredient and medications according to therapeutic actions and active compounds. RESULTS: 24.3% of the athletes tested for doping control declared no use of medications or food supplements. Food supplements (45.3%) continue to be popular, with vitamins (43.2%) and proteins/aminoacids (13.9%) in power sports being most widely used. Nonsteroidal antiinflammatory agents and analgesics were also commonly used by athletes (11.1% and 3.7%, respectively). The use of the hemoderivative actovegin and several nonprohibited anabolic preparations are discussed. The prevalence of medication use for asthma and the dangers of drug interactions are also presented.Laboratory analysis data reveal that of the aTUEs received for inhaled glucocorticosteroids, only budesonide was detectable in significant percentage (10.0%). Only 6.5% of the 445 athletes approved to inhale beta2-agonists led to an adverse analytical finding. CONCLUSIONS: This review demonstrates that overuse of food supplements was slightly reduced compared to previous OGs and a more rational approach to the use of medication is being adopted. PMID- 19124983 TI - Does the use of fluoroscopy and isometry during anterior cruciate ligament reconstruction affect surgical decision making? AB - OBJECTIVE: Poor results after anterior cruciate ligament (ACL) reconstruction are often due to inaccurate graft placement. Numerous strategies have been advocated to improve accuracy and consistency of tunnel positioning, including computer assisted navigation. Less expensive alternatives, such as intraoperative fluoroscopy and isometry, have also been advocated for confirming guide pin placement before reaming the femoral tunnel. It is unknown how often these techniques cause surgeons to change the location of their femoral tunnel at the time of surgery. We undertook this study to determine how often this approach results in repositioning of the guide pin before final graft placement. We hypothesized that a lower level of surgeon experience would lead to a higher frequency of repositioning compared to a higher level of experience. DESIGN: Prospective, case series. SETTING: Institutional. PATIENTS: Intraoperative data were gathered prospectively from 413 consecutive, primary ACL reconstructions performed by the sports medicine group at our institution. Of the 413 procedures enrolled in this study, 407 were available for analysis. Six procedures were excluded because the tension isometer was unavailable during the procedure. INTERVENTIONS: Isometry and fluoroscopy were used in all cases to aid in the accurate placement of the femoral tunnel. Femoral pin change based on the results of isometry or fluoroscopy was recorded. The percentage of cases involving a change in the femoral pin resulting from the use of these techniques was calculated. This percentage was also calculated separately for cases performed by a staff surgeon (fellowship-trained sports medicine staff) as well as less experienced surgeons (current sports medicine fellows). MAIN OUTCOME MEASURES: The main outcome measurement was whether the femoral pin was changed. RESULTS: Of the 407 procedures available for review, 62 (15%) of them involved a change in femoral pin position secondary to information provided by intraoperative isometry or fluoroscopy. In the procedures performed by more experienced surgeons, the pin was changed in 40 (16%) of 253 cases; in those performed by less experienced surgeons, it was changed in 22 (14%) of 154 cases. CONCLUSIONS: The intraoperative use of isometry and fluoroscopy during ACL reconstruction led to changes in the femoral tunnel placement 15% of the time. The influence of these instruments on intraoperative decision making does not seem to diminish with surgical experience. PMID- 19124982 TI - Resting energy expenditure: a stronger marker than body weight for bone mineral density in white women but not men? The Rancho Bernardo study. AB - OBJECTIVE: To investigate the association between resting energy expenditure (REE) and bone mineral density (BMD) in white adults. DESIGN: Cross-sectional study. SETTING: White, middle-class to upper middle-class community-dwelling adults. PARTICIPANTS: Women (n = 996) and men (n = 686) aged 36 to 97 years. ASSESSMENT OF RISK FACTORS: REE calculated using the Harris-Benedict equation. MAIN OUTCOME MEASURES: BMD of the lumbar spine (L1 to L4), total hip, and total body measured using dual energy x-ray absorptiometry. RESULTS: REE was lower in women than in men (1220 versus 1566 kcal/day, P < 0.0001); women also had lower BMI. After adjusting for lean body mass (LBM), REE was higher in women than men (1407 versus 1296 kcal/kg LBM/d, P < 0.0001). In stepwise multiple linear regression models, REE explained 13% of variance in spine BMD in women and 6% in men, 33% of variance in hip BMD in women and 22% in men, and 32% of variance in total body BMD in women and 22% in men. In women, weight explained 10% less of the hip BMD variance and 6% less of the total body BMD variance than REE. In men, weight explained 4% more of the spine BMD variance and 1% more of the total body BMD variance than REE. CONCLUSIONS: REE explained more of the BMD variance than weight in women, and the reverse was true in men. These sex differences were largely explained by sex differences in LBM or weight. PMID- 19124984 TI - Pilot study of a 10-week multidisciplinary Tai Chi intervention in sedentary obese women. AB - OBJECTIVE: Alternative approaches to weight control and physical activity are increasingly needed. Numerous factors influence weight management, including the choice of physical exercise. No study has previously examined the therapeutic effect of a multidisciplinary weight management program incorporating Tai Chi (TC) exercises among sedentary obese women. DESIGN: Randomized intervention trial with blinded medical provider. SETTING: In day hospital consultations. PARTICIPANTS: Twenty-one obese women. INTERVENTION: All subjects participated in a 10-week weight management program that was part of usual care and included a hypocaloric balanced diet, a weekly physician/psychologist/dietician group session, and an exercise program. For the exercise component, subjects were randomized to either a 2-hour weekly session of TC or a conventional structured exercise program. MAIN OUTCOME MEASURES: Changes in weight, body composition, heart rate, blood pressure, mobility scores, mood, Three Factor Eating Questionnaire scores, and General Self-Efficacy. RESULTS: The TC arm improved in resting systolic blood pressure, chair rise test, mood, and reduced percent of fat at week 10 and at 6 months follow-up. General self-efficacy was enhanced in both groups and maintained at 30 weeks. CONCLUSION: The observed benefits over a 30-week period of a multidisciplinary weight management program incorporating TC exercises on physical functioning mood and dietary restraint need further understanding of how sedentary obese women adhere to physical activity like TC or other alternative exercises. PMID- 19124985 TI - Nonoperative treatment of midportion Achilles tendinopathy: a systematic review. AB - OBJECTIVE: The aim of this systematic review is to provide an easily accessible, clear summary of the best available evidence for nonoperative treatment of midportion Achilles tendinopathy. DATA SOURCES: MEDLINE, CINAHL, and Embase through April 2007. Search terms: achilles tendon or tendo achilles or triceps surae or tendoachilles or tendo-achilles or achilles AND tendinopathy or tendinosis or tendonitis or tenosynovitis. STUDY SELECTION: Of 707 abstracts reviewed, 16 randomized trials met our inclusion criteria. DATA EXTRACTION: Data extracted from each paper included: patient demographics (age and sex), duration of symptoms, method of diagnosis, treatments, cohort size, length of follow-up, pain-related outcome data, and secondary outcome data. DATA SYNTHESIS: The primary outcome measurement was change in numeric pain score. Focal tenderness, tendon thickness, and validated outcome scores were used secondarily. Eccentric exercises were noted to be equivalent to extracorporeal shockwave therapy (1 study) and superior to wait-and-see treatment (2 trials), traditional concentric exercise (2 of 3 trials), and night splints (1 study). Extracorporeal shockwave therapy was shown to be superior to a wait-and-see method in 1 study but not superior to placebo in another. Sclerosing injections were shown to be superior to placebo in 1 study, but local steroid treatment was beneficial in 2 of 3 studies. Injection of deproteinized hemodialysate and topical glyceryl nitrate application were beneficial in 1 trial each. CONCLUSIONS: Eccentric exercises have the most evidence of effectiveness in treatment of midportion Achilles tendinopathy. More investigation is needed into the utility of extracorporeal shockwave therapy, local corticosteroid treatments, injections of sclerosing agents or deproteinized hemodialysate, and topical glyceryl nitrate application. PMID- 19124986 TI - First rib stress fracture and pseudarthrosis in the adolescent athlete: the role of costosternal anatomy. PMID- 19124987 TI - Traumatic dislocation of tibialis posterior tendon: a case report in a Tae-Kwon Do athlete. PMID- 19124988 TI - Hepatic injury after costochondral separation in a rugby football player. PMID- 19124989 TI - A successful therapy of high-altitude pulmonary edema with a CPAP helmet on Lenin Peak. PMID- 19124991 TI - Aberrant EBV antibody patterns may mimic acute EBV infections. PMID- 19124992 TI - Effects of growth hormone on athletic performance: a review. PMID- 19124993 TI - Open versus closed kinetic chain exercises for patellar chondromalacia in young women. PMID- 19124994 TI - Prevention of running-related injuries among novices. PMID- 19124995 TI - Lower extremity injuries in non-elite runners: a review. PMID- 19124997 TI - Current developments for pelvic exenteration in gynecologic oncology. AB - PURPOSE OF REVIEW: The present review aims to update new techniques of pelvic exenteration including minimal invasive surgery, and discuss other aspects of this radical surgery, including worldwide differences. RECENT FINDINGS: Major advances are made since the first description of pelvic exenteration and the operation is still under evolution. Explorative laparoscopy prior to exenteration is a valuable alternative to laparotomy to elect candidates for pelvic exenteration. There are considerable differences with respect to indications, contraindications, preoperative staging and adjuvant therapy after exenteration in different countries. Advances in laparoscopic instruments also led to the laparoscopic exenteration. The main limiting step of the operation is urinary diversion. New techniques of laparoscopic-assisted and robotic-assisted techniques of urinary diversion have been reported that decrease the operation time. Vascularized muscle flaps are preferred by many surgeons to fill the empty pelvis and provide an acceptable vaginal reconstruction. J-pouch seems to be a safer technique than end-to-end coloanal anastomosis for bowel reconstruction. Developments in the bioengineering tissue for pelvic reconstruction are required. SUMMARY: Laparoscopy has the advantages of decreased blood loss, improved convalescence, lower incidence of wound infection and incisional hernia, short recovery periods, rapid return of bowel function, better pain control and improved cosmetics compared with laparotomy for pelvic exenteration. Magnification and improved visualization permits en-bloc dissection of tumor and good anastomosis technique. New techniques of urinary diversion, orthotopic neobladder and coloanal are promising. PMID- 19124998 TI - Role of complete para-aortic lymphadenectomy in endometrial cancer. AB - PURPOSE OF REVIEW: Many reports suggest the significance of pelvic lymph-node (PLN) adenectomy in patients with endometrial cancer. However, among these, there is controversy regarding not only what type of patients should have lymphadenectomy performed, but also what extent lymphadenectomy should be performed. RECENT FINDINGS: It has been reported that PLN adenectomy has therapeutic significance in stage I grade 3 and more advanced endometrioid uterine cancer. However, the effects of para-aortic lymph-node adenectomy on its prognostic benefit have not been discussed. SUMMARY: Patients with low-risk disease might not benefit from PLN adenectomy. However, PLN adenectomy might still have merit in low-risk patients, as there are inaccuracies of preoperative and intraoperative assessments. A complete lymphadenectomy is safe with minimum complications. At this point, hysterectomy and bilateral salpingo-oophorectomy with complete PLN adenectomy as the standard surgical procedure for endometrial cancer is thought to be reasonable. At present, the addition of p-aortic lymph node adenectomy is regarded as an investigated protocol for endometrial cancer. However, p-aortic lymph-node adenectomy may have a therapeutic role for stage IIIC patients. Prospective randomized controlled trial composed of intermediate/high-risk patients should be conducted to clearly demonstrate prognostic improvement by p-aortic lymph-node adenectomy itself. PMID- 19124999 TI - Cytoreductive surgery and perioperative intraperitoneal chemotherapy for the treatment of advanced primary and recurrent ovarian cancer. AB - PURPOSE OF REVIEW: With a steady incidence in the United States and worldwide, ovarian cancer remains the number one cause of death from gynecological malignancies. The conventional treatment approach for advanced ovarian malignancy is surgical debulking and systemic chemotherapy. More than 50% of these patients relapse within 5 years. RECENT FINDINGS: For advanced primary and recurrent disease, a comprehensive approach including cytoreductive surgery and perioperative intraperitoneal chemotherapy, based on the experience with other peritoneal surface malignancies is proposed. The goal of these treatments is to surgically eradicate all visible tumor and to chemically and physically eradicate microscopic residual disease. Cytoreductive surgery includes peritonectomy procedures and visceral resections. At the Washington Cancer Institute, cisplatin and doxorubicin are administered through the intraperitoneal route with heat during the surgical procedure. In the first 5 postoperative days, patients receive normothermic intraperitoneal paclitaxel. SUMMARY: Results of several phase II trials with this comprehensive approach suggested improved survival as compared with historical controls with reasonable morbidity and mortality. PMID- 19125000 TI - Robotic surgery in gynecologic oncology. AB - PURPOSE OF REVIEW: Robotic surgery is rapidly taking the place of laparoscopy in many gynecologic oncology practices. Numerous practitioners have published their experience with this new technology. A review of their findings is timely and relevant. RECENT FINDINGS: The majority of case series of robotic surgery for hysterectomy and lymphadenectomy show that the procedure is feasible and at least comparable to laparoscopic surgery. Similarly, case series of robotic radical hysterectomy for cervical cancer also compare favorably to laparoscopic surgery. Less common procedures such as robotic trachelectomy, parametrectomy, and retroperitoneal lymphadenectomy have also been described. Numerous patient and practitioner advantages are discussed in this review. SUMMARY: Robotic surgery is a minimally invasive alternative to laparoscopy for the surgical treatment of endometrial cancer and cervical cancer. Its role in ovarian cancer is just starting to be explored. PMID- 19125001 TI - Hereditary non-polyposis colorectal cancer or Lynch syndrome: the gynaecological perspective. AB - PURPOSE OF REVIEW: Hereditary non-polyposis colorectal cancer (HNPCC) or Lynch syndrome is characterized by a number of other cancers including colorectal, endometrial and ovarian cancer. This review covers the gynaecological aspects of managing women with HNPCC: diagnostic criteria, molecular tests for diagnosis, cancer risks and different strategies for surveillance and prevention. RECENT FINDINGS: Studies correcting for ascertainment bias found slightly lower penetrance estimates than those obtained from high-risk families. HNPCC linked ovarian cancer presents at an earlier age and stage and has similar survival rates as sporadic cancer. In endometrial tumours, microsatellite instability or immunohistochemistry has limited effectiveness in selecting individuals for genetic testing, due to molecular differences. Population-based data indicate that a significant proportion of mismatch repair gene carriers would be missed by current clinical criteria. Effective risk prediction models complement clinical risk assessment. Effectiveness of screening is unproven and prophylactic surgery is the best preventive option for women who have completed their families. Multimodal screening protocols from the age of 30-35 years are being evaluated. SUMMARY: Risk of endometrial cancer in women with Lynch syndrome is as high as the risk of colorectal cancer. Further research is needed to identify the appropriate strategy for clinical risk assessment and optimize screening. A multidisciplinary approach is necessary to manage these women. PMID- 19125002 TI - Chemosensitivity and chemoresistance testing in ovarian cancer. AB - PURPOSE OF REVIEW: Sensitivity testing in ovarian cancer to individualize therapy remains an active area of interest and this has been renewed recently by results from several groups. The clinical results of assay-directed therapy are invariably better than would be expected from empirical treatment, but it has proved difficult to get these tests into practice. RECENT FINDINGS: Several recent studies suggest that cellular individualized tumour response tests, particularly the ATP-based tumour chemosensitivity assay, can improve the chance of response and probably survival for individual patients. Most tumour response tests show excellent correlation with clinical resistance, but vary in their ability to predict sensitivity. Molecular assays of sensitivity and resistance are less developed in ovarian cancer than in breast cancer, but those using multiple gene signatures show considerable promise. SUMMARY: Individualized therapy has the ability to guide the use of chemotherapy as well as newer agents. The development of companion diagnostics for drugs used in ovarian cancer has considerable potential for the future and such assays are already proving useful where there is clinical evidence of equivalence between possible treatments. PMID- 19125003 TI - New therapeutic agents in ovarian cancer. AB - PURPOSE OF REVIEW: Despite advances in management over recent years, epithelial ovarian cancer remains the most lethal gynaecological malignancy. Methods of early detection, as well as improved therapeutic options, are urgently needed. RECENT FINDINGS: Currently, a number of targeted therapies, including vascular endothelial growth factor inhibitors, poly-ADP-ribose polymerase inhibitors and folate receptor inhibitors look promising in this arena and this article will review a number of these drugs and the evidence pertaining to their use. SUMMARY: Much further research is required to define if, when and how best to integrate these novel therapies, and also to define associated biomarkers that predict toxicity and select patients most likely to derive benefit. Individualized therapy is not an impossible dream, but there is still a long way to go. PMID- 19125004 TI - Host immune responses to cervical cancer. AB - PURPOSE OF REVIEW: The purpose of this review is to understand the role of the host immune system in clearing the human papillomavirus (HPV) infection, strategies adopted by HPV to subvert the host immune responses and analyze the challenges to the future immunotherapeutic treatment modalities. RECENT FINDINGS: Cervical epithelium provides a protective niche to the virus to subvert the immune responses. The absence of an inflammatory milieu in the cervix makes the resident dendritic and langerhan cells tolerogenic to HPV antigens. CD4 cells predominated in regressing cervical intraepithelial neoplasia lesions, whereas CD8 cells were dominant in invasive carcinoma. A reduced expression of T cell signaling molecule T-cell receptor zeta chain was observed in CD8 lymphocytes. Decreased numbers of NKG2D expressing natural killer and T cells were present in patients with cervical cancer and cervical intraepithelial neoplasia. Increased frequencies of CD4 CD25+ FoxP3+ T regulatory cells were observed in patients with cervical cancer. The Nrp-1+Treg showed greater suppressive activity. A network of Treg and indoleamine 2, 3-dioxygenase expressed in tumor cells facilitates immune escape of tumor cells. SUMMARY: The HPV uses different strategies to evade immune recognition. Understanding the immune evasion mechanisms used by HPV will help in designing newer therapeutic strategies for cervical cancer. PMID- 19125005 TI - Aromatase inhibitors and bone health. AB - PURPOSE OF REVIEW: Aromatase inhibitors improve survival in breast cancer patients but adversely affect bone health. RECENT FINDINGS: Hormone receptor positive breast cancer is increasingly targeted with chemotherapy, ovarian suppression and the use of aromatase inhibitors. Aromatase inhibitors block oestrogen production in peripheral tissues and the three third generation aromatase inhibitors (anastrozole, letrozole and exemestane) reduce circulating oestrogen levels, leading to accelerated bone loss and an increased risk of fracture.The majority of fractures occur in osteopaenic women prescribed aromatase inhibitors. Current guidelines advocate bone mineral density (BMD) measurement in all patients on aromatase inhibitors with selective use of antiresorptive therapy in osteoporotic (T-score > -2.5) women. Risk factors for premature bone loss and fracture include a low BMI, family history or personal history of fragility fracture after the age of 50, oral corticosteroid use more than 6 months and cigarette smoking. Emerging evidence supports concomitant use of bisphosphonates in all women on aromatase inhibitors to prevent fracture and breast cancer recurrence. SUMMARY: The increasing use of aromatase inhibitors requires selection of patients for antiresorptive therapy and careful bone health management to reduce bone loss and prevent fragility fractures. PMID- 19125006 TI - Techniques for fertility preservation in patients with breast cancer. AB - PURPOSE OF REVIEW: To outline the risks of infertility from breast cancer treatment, and to illustrate current techniques in preserving fertility in breast cancer patients who wish to become pregnant after treatment is concluded. RECENT FINDINGS: Breast cancer often affects women of reproductive age. Although treatment is effective, cytotoxic chemotherapy causes ovarian reserve depletion, whereas hormonal therapy necessitates a delay in pregnancy, resulting in infertility. Patients of reproductive age should be referred to fertility specialists to explore methods of fertility preservation upon diagnosis. The best established method of fertility preservation is embryo cryopreservation, although investigational techniques such as, oocyte and ovarian tissue cryopreservation, may hold potential. Embryo cryopreservation involves ovarian stimulation to retrieve oocytes in-vitro fertilization prior to freezing. Techniques for the cryopreservation of unfertilized oocytes are under investigation. Successful pregnancies have resulted in breast cancer patients after treatment, without obvious compromise in their risk of recurrence or death from breast cancer. SUMMARY: Ovarian stimulation with retrieval of ooctyes for in-vitro fertilization remains the best known option for fertility preservation in women with early stage breast cancer whose risk of fertility may be compromised by adjuvant chemotherapy. PMID- 19125007 TI - Impact of diet on breast cancer risk. AB - PURPOSE OF REVIEW: Breast cancer is the most common female cancer in Western Europe and North America, and becoming an increasing problem in developing countries such as India and China. We review recent studies (published 1 January 2007-31 August 2008) on the impact of diet on breast cancer risk. RECENT FINDINGS: Recent studies have focused on the controversial association for dietary fat and breast cancer as well as the role of newer aspects such as glycemic index, dietary patterns and diet-gene interactions. Evidence that some of the associations may be modified by oestrogen and progesterone receptor status has been presented. Still, only alcohol intake, being overweight and weight gain have shown consistent and strong positive associations with breast cancer risk. The reasons for the null or weak associations often observed regarding diet and breast cancer might be several. For example, there may be no causal association, or existing associations may be masked by measurement error, timing of dietary exposure and differences according to tumour characteristics or diet-gene interactions. SUMMARY: Numerous epidemiological studies on diet and breast cancer have been published during our review period. Still, only alcohol intake, being overweight and weight gain have shown consistent and strong positive associations with breast cancer risk. PMID- 19125008 TI - Urogynaecological problems in pregnancy and postpartum sequelae. AB - PURPOSE OF REVIEW: Women in pregnancy can experience lower urinary tract symptoms which are related to the pregnancy and delivery and iatrogenic, and related to use of epidural anaesthesia and urethral catheters. This article assesses the controversial relationship between pregnancy and delivery and the development of urinary incontinence and pelvic organ prolapse. RECENT FINDINGS: Lower urinary tract symptoms are common in pregnancy and they peak in the third trimester. If women have lower urinary tract symptoms prior to pregnancy, they are more likely to persist after delivery. Vaginal delivery is the factor most strongly associated with stress urinary incontinence after delivery and elective caesarean section may be protective. Vaginal childbirth causes levator ani injury and increase in levator hiatus size, and these persist following vaginal delivery. Women with levator ani injuries may be twice as likely to develop uterovaginal prolapse. Voiding difficulties are more likely to occur after a traditional epidural than a low dose or combined spinal epidural. SUMMARY: There is radiological evidence supporting pelvic floor injury following vaginal childbirth and epidemiological evidence for the relationship between vaginal delivery and urinary incontinence and pelvic organ prolapse. Rigorous long-term studies are needed to identify the direct relationship between these two phenomena. PMID- 19125009 TI - Current world literature. PMID- 19125011 TI - An update on papillary microcarcinoma. AB - PURPOSE OF REVIEW: There is a worldwide increasing incidence of thyroid cancer, mostly due to the increased diagnosis of papillary microcarcinoma (PMC). This review examines its clinical significance and optimal treatment. RECENT FINDINGS: It is unclear whether or not the increase in PMC is real, or is simply due to the increased detection and diagnosis. The increase has been attributed to the widespread availability of thyroid ultrasound, with increasing numbers of clinically insignificant thyroid nodules subject to fine needle aspiration biopsy and surgical excision. An additional explanation is simply an increase in the number of routine histologic sections taken from the thyroidectomy specimens over the years. There have been two recent large clinical series with long-term follow up highlighting the excellent prognosis of PMC. Novel research into immunohistochemical markers and advanced imaging with fluorodeoxyglucose positron emission tomography aims to identify the markers of more aggressive PMC. Management of PMC remains controversial, from observation alone to total thyroidectomy/radioiodine ablation. SUMMARY: PMC has been increasing rapidly over recent decades due to increased cervical ultrasound and increased detection in pathological specimens. There is no compelling evidence base for the management of PMC, with retrospective series highlighting the excellent prognosis. PMID- 19125012 TI - Medullary thyroid cancer: early detection and novel treatments. AB - PURPOSE OF REVIEW: Medullary thyroid cancer (MTC) is derived from the parafollicular cells of the thyroid. Understanding the molecular biology behind specific mutations of the RET gene and their prognostic implications have led to the establishment of tailored treatment modalities for certain patients. We review the most recent studies on the molecular biology, calcitonin screening, diagnosis, imaging, and treatment of MTC. RECENT FINDINGS: Newly identified rearranged during transfection point mutations have helped with MTC prognosis and have resulted in the establishment of new treatment guidelines. Screening for MTC in the United States with basal serum calcitonin for patients with thyroid nodules would cost $11,793 per life-year saved (LYS), compared with colonoscopy and mammography screening. For metastatic or recurrent disease, neck ultrasound, chest computed tomography scan, liver MRI, bone scintigraphy, and axial skeleton MRI have been proven superior to 18F-FDG PET/computed tomography. For patients with nonoperable metastatic disease, novel chemotherapeutic agents, such as vandetanib, targeting rearranged during transfection, vascular endothelial growth factor receptor and epidermal growth factor receptor, are showing promise. Such agents are currently in phase II trials. SUMMARY: There have been several recent advances in the diagnosis, molecular biology, imaging, and treatment options of MTC. By potentially downstaging of disease, and treating metastatic disease more effectively, overall survival and outcomes of patients may improve. PMID- 19125013 TI - Recent developments in predicting thyroid malignancy. AB - PURPOSE OF REVIEW: To provide an update on potential predictors of thyroid malignancy and how their use may alter clinical management. RECENT FINDINGS: As thyroid nodules become more prevalent clinicians are increasingly impelled to identify the optimal predictor(s) of thyroid malignancy, with the goal of guiding management based on assessed risk. The gold standard evaluation for thyroid nodules is ultrasound-guided fine-needle aspiration biopsy. Fine-needle aspiration biopsy is not perfect and adjuncts which complement its predictive value are being investigated from several innovative perspectives. These include large tumor size (> or =4 cm), which appears to be an independent predictor of thyroid malignancy; 18F-fluorodeoxyglucose positron emission tomography, which appears to facilitate exclusion of malignancy in cytologically indeterminate thyroid nodules; and peripheral blood and fine-needle aspiration biopsy analysis of molecular markers, which may help to identify malignant thyroid nodules with greater specificity. SUMMARY: Fine-needle aspiration biopsy of large thyroid nodules has a high false-negative rate and should be considered for diagnostic lobectomy. Nodule size appears to be an independent factor predicting malignancy and indeterminate lesions at least 4 cm should be considered for initial total thyroidectomy. Nuclear imaging may aid exclusion of malignancy in thyroid nodules and molecular markers have great promise in predicting thyroid malignancy with higher specificity. PMID- 19125014 TI - Predictors of outcomes following pediatric thyroid and parathyroid surgery. AB - PURPOSE OF REVIEW: Recent studies have demonstrated racial/ethnic and socioeconomic disparities in adults undergoing thyroidectomy and parathyroidectomy. To date, few studies have examined outcomes in children undergoing cervical endocrine surgery. RECENT FINDINGS: Children undergoing thyroidectomy and parathyroidectomy have higher complication rates than adults undergoing similar procedures. Complication rates appear to be lower when procedures are performed by high-volume surgeons. Access to high-volume surgeons continues to be limited for children of racial/ethnic minorities and in families of a lower socioeconomic status. SUMMARY: Complications following thyroidectomy and parathyroidectomy in children can have profound, life-long effects on development and quality of life. Outcomes are optimized when surgery is performed by high-volume surgeons. Obtaining access to high-volume surgeons requires a multidisciplinary approach by parents, pediatricians, pediatric endocrinologists, and third-party payers. PMID- 19125015 TI - Molecular genetics of gastroenteropancreatic neuroendocrine tumors. AB - PURPOSE OF REVIEW: Gastroenteropancreatic neuroendocrine tumors (GEP NETs) are relatively rare neoplasias arising from the embryonic neural crest, neuroectoderm and endoderm. GEP NETs occur either sporadically or as part of endocrine tumor susceptibility syndromes such as multiple endocrine neoplasia type 1 (MEN1), von Hippel Lindau (VHL) syndrome, neurofibromatosis (NF-1), and possibly tuberous sclerosis (TSC). The overall incidence of GEP NETs shows a significant increase over the past three decades. Improved understanding of the molecular genetics associated with these lesions will likely enhance the diagnosis and treatment of patients with GEP NET. RECENT FINDINGS: The molecular and clinical genetics of familial GEP NETs have been further elucidated by the characterization of the tumor suppressor genes, MEN1, VHL, NF-1, TSC1, and TSC2. The vastly improved technology in the field of cancer genetics with higher resolution of the study of genetic alterations, and the ability of unbiased mutational analyses of entire tumor genomes is likely to further the understanding of the genetic mechanisms of sporadic GEP NET as well. SUMMARY: Recent advances in the molecular genetics of sporadic and familial GEP NET are reviewed. PMID- 19125016 TI - Management of adrenal lesions. AB - PURPOSE OF REVIEW: Adrenal lesions are commonly seen on modern imaging modalities, and although the majority are benign, potentially lethal entities necessitate exclusion. This group of both functional lesions and malignancies can produce an extensive variety of symptoms. The purpose of this review is to summarize recent advances in diagnosis and therapies for adrenal lesions. RECENT FINDINGS: New tumor markers and genetic risk factors continue to be discovered, and improved diagnostic techniques have made adrenal incidentalomas more common than ever before. Laparoscopic approaches for adrenal lesions continue to evolve for functional lesions, and also for lesions 12 cm and larger. Open adrenalectomy continues to be the most appropriate for adrenocortical cancer. SUMMARY: All adrenal lesions should prompt a functional hormonal assessment and additional imaging to determine malignancy employed. PMID- 19125017 TI - Cancer stem cells: never Wnt away from the niche. AB - PURPOSE OF REVIEW: The last years of cancer research have established the concept of cancer stem cells (CSCs) as a subpopulation of cells within a tumor entirely responsible for tumorigenesis. This has aroused expectations that targeting cancer stem cells would allow effective tumor eradication. This review aims to summarize the relevant achievements in the field and to highlight the complex mechanisms that are involved in regulating CSC function. RECENT FINDINGS: A growing number of studies have identified CSCs in a variety of human tumor types. The focus of attention is now moving to discover molecular signals which are essential to sustain CSC. We summarize recent results on intrinsic and extrinsic signaling pathways such as Wnt signals, which control stem cell self-renewal and highlight the role of the microenvironment or niche in this process. SUMMARY: The discovery of cancer stem cells points into new directions to gain better understanding of cancer biology. This is expected to alter the design of clinical research programs and to improve the way we assess the efficacy of novel anticancer drugs. Several conclusions and predictions derived from this concept hold great promise to speed up the process of discovering effective targets for clinical application. PMID- 19125018 TI - Sequence-based advances in the definition of cancer-associated gene mutations. AB - PURPOSE OF REVIEW: Recent rapid progress in DNA sequencing has permitted projects to be undertaken that are aimed at building unbiased genome-wide portraits of the underlying mutations in human tumors. This review sets out the highlights of the recent progress in this area and the rapidly evolving picture of the underlying genetic basis of human epithelial cancers. RECENT FINDINGS: Individual tumors are estimated to contain around 80 point mutations in protein coding genes of which 15 are likely to be tumorigenic. It is likely that there are hundreds of different genes that when mutated contribute to human tumorigenesis most in only a small fraction of tumors. Mutations caused by large chromosomal rearrangements also appear to be common in tumors. In prostate and lung cancers, recurrent chromosomal translocations resulting in tumorigenic fusion proteins have been identified. SUMMARY: The multitude of new mutated genes being identified in human tumors represent many new directions for experimental research into the molecular pathways that lead to tumor formation. These studies, in turn, are likely to lead to many novel approaches to targeted therapy useful in subsets of tumors with particular types of gene mutation. PMID- 19125020 TI - Corrupt policemen: inflammatory cells promote tumor angiogenesis. AB - PURPOSE OF REVIEW: The last decade has seen a paradigm change in that tumor stroma contributes to malignant tumor progression in a manner comparable to genetic and epigenetic changes within cancer cells. This review summarizes recent novel insights into how inflammatory conditions stimulate the formation and expansion of blood and lymphatic vessels within tumors and, thus, allow tumors to grow, to gain invasive capabilities, and to finally seed metastasis in distant organs. RECENT FINDINGS: Different cancer types have a highly defined microenvironment, which is composed of cancer-associated fibroblasts, blood vessel and lymphatic endothelial cells, pericytes, and a heterogeneous infiltrate of cells of the immune system. In addition to the local stimulation of tumor angiogenesis and tumor lymphangiogenesis, cytokines released by the primary tumor and by the immune cell infiltrate also instruct bone marrow-derived cells to colonize distant organs and to prepare these sites for future metastasis. SUMMARY: Inflammatory reactions coinciding with carcinogenesis can be visualized by the presence of specific bone marrow-derived, inflammatory cells in patients' peripheral blood. Recent findings suggest that such inflammatory fingerprints may better define the inflammatory nature of the primary malignancy and, thus, allow the design of therapeutic strategies targeting the protumorigenic immune cell stroma compartment. PMID- 19125019 TI - Behavior of immune players in the tumor microenvironment. AB - PURPOSE OF REVIEW: Tumors recruit various immune cells with seemingly contrasting functions. Yet, the precise role of these cells in situ remains vastly unknown. This review presents a new discovery effort that employs intravital imaging to study immune players directly in tissues. RECENT FINDINGS: Cytotoxic T lymphocytes (CTLs) that recognize cognate antigenic peptide can infiltrate tumors from the periphery to the center, and physically engage and eliminate antigen presenting tumor cells. Nevertheless, the reported kinetics for tumor cell killing by CTLs in vivo is surprisingly low as it takes several hours for one CTL to eliminate one tumor cell. Also, T regulatory (Treg) cells can create a suppressive milieu that restricts the release of CTL cytotoxic granules, which protects tumor cells from being killed. CTLs may be further subverted during lengthy interactions with tumor-associated macrophages (TAMs). Finally, TAMs can directly facilitate tumor invasion by recruiting tumor cells nearby vessels and promoting their intravasation. SUMMARY: Intravital imaging has started to uncover tumor-related immune events as they unfold in vivo. The technology should be exploited in the coming years to dissect further the tumor microenvironment and to define therapeutics that augment antitumor immunity. PMID- 19125021 TI - The immunogenicity of tumor cell death. AB - PURPOSE OF REVIEW: It is an ongoing conundrum under which circumstances cellular demise induces an immune response and whether apoptotic or necrotic cells are intrinsically immunogenic or tolerogenic. This review summarizes recent insights in the immunogenicity of dying tumor cells. RECENT FINDINGS: Although apoptosis appears to be morphologically homogeneous, recent evidence suggests that the preapoptotic surface exposure of calreticulin (CRT) may have a profound impact on the immune response. Moreover, the release of high mobility group box 1 protein (HMGB1) during late apoptosis promotes antigen processing by dendritic cells and hence contributes to efficient antigen presentation and cytotoxic T-cell activation. HMGB1 is sensed by TLR4 on dendritic cells, and loss-of-function alleles of TLR4 abolish anticancer immune response and accelerate tumor progression. SUMMARY: A combination of signals elicits an efficient immune response against tumor cells that are dying in response to anthracyclines or ionizing irradiation. During early apoptosis, caspase activation and endoplasmic reticulum stress facilitate the surface exposure of immunogenic effectors followed by the release of soluble factors that are indispensable for effective immune response. Failure of tumor cells to expose/secrete such immunogenic factors and/or failure of the immune system to sense such effectors may compromise the efficacy of conventional anticancer therapies. PMID- 19125022 TI - Bone marrow-derived cells and epithelial tumours: more than just an inflammatory relationship. AB - PURPOSE OF REVIEW: Cancer-associated fibroblasts/myofibroblasts and inflammatory cells produce a vast array of growth factors, chemokines and extracellular matrix (ECM) components that facilitate cancer progression, invasion/metastasis and neovascularization. This review highlights some surprisingly novel mechanisms of this paracrine relationship. RECENT FINDINGS: Mesenchymal stem/stromal cells (MSCs) are known for their tropism towards certain tumours, but now we find that cross-talk between tumours and MSCs leads to greater tumour motility and metastasis. Two closely related populations of immature myeloid cells, so-called 'cap cells' and myeloid-derived suppressor cells (MDSCs) also cross-talk with tumour cells, promoting invasion and metastasis through matrix metalloproteinase (MMP) secretion, as well as contributing to neovascularization and T-cell tolerance. The contribution of bone marrow-derived cells (BMDCs) to tumour neovascularization is controversial, but BMD--endothelial progenitor cells (EPCs) -are strongly implicated in the angiogenic switch in a mouse model. BMDCs are also credited with the creation of premetastatic niches to which metastatic cells adhere via integrins. SUMMARY: There is no doubt that BMDCs are not simply bystanders in the tumour battleground. The mechanisms through which they aid tumour progression are numerous; effective treatments that combat BMDC-tumour cross-talk are surely on the way. PMID- 19125023 TI - Current world literature. PMID- 19125024 TI - Prevalence of GJB2 (connexin-26) and GJB6 (connexin-30) mutations in a cohort of 300 Brazilian hearing-impaired individuals: implications for diagnosis and genetic counseling. AB - OBJECTIVE: Hereditary nonsyndromic deafness is an autosomal recessive condition in about 80% of cases, and point mutations in the GJB2 gene (connexin 26) and two deletions in the GJB6 gene (connexin 30), del(GJB6-D13S1830) and del(GJB6 D13S1854), are reported to account for 50% of recessive deafness. Aiming at establishing the frequencies of GJB2 mutations and GJB6 deletions in the Brazilian population, we screened 300 unrelated individuals with hearing impairment, who were not affected by known deafness related syndromes. METHODS: We firstly screened the most frequently reported mutations, c.35delG and c.167delT in the GJB2 gene, and del(GJB6-D13S1830) and del(GJB6-D13S1854) in the GJB6 gene, through specific techniques. The detected c.35delG and c.167delT mutations were validated by sequencing. Other mutations in the GJB2 gene were screened by single-strand conformation polymorphism and the coding region was sequenced when abnormal patterns were found. RESULTS: Pathogenic mutations in GJB2 and GJB6 genes were detected in 41 individuals (13.7%), and 80.5% (33/41) presented these mutations in homozygosis or compound heterozygosis, thus explaining their hearing defect. The c.35delG in the GJB2 gene was the most frequent mutation (37/300; 12.4%), detected in 23% familial and 6.2% the sporadic cases. The second most frequent mutation (1%; 3/300) was the del(GJB6-D13S1830), always found associated with the c.35delG mutation. Nineteen different sequence variations were found in the GJB2 gene. In addition to the c.35delG mutation, nine known pathogenic alterations were detected c.167delT, p.Trp24X, p.Val37Ile, c.176_191del16, c.235delC, p.Leu90Pro, p.Arg127His, c.509insA, and p.Arg184Pro. Five substitutions had been previously considered benign polymorphisms: c.-15C>T, p.Val27Ile, p.Met34Thr, p.Ala40Ala, and p.Gly160Ser. Two previously reported mutations of unknown pathogenicity were found (p.Lys168Arg, and c.684C>A), and two novel substitutions, p.Leu81Val (c.G241C) and p.Met195Val (c.A583G), both in heterozygosis without an accompanying mutation in the other allele. None of these latter four variants of undefined status was present in a sample of 100 hearing controls. CONCLUSIONS: The present study demonstrates that mutations in the GJB2 gene and del(GJB6 D13S1830) are important causes of hearing impairment in Brazil, thus justifying their screening in a routine basis. The diversity of variants in our sample reflects the ethnic heterogeneity of the Brazilian population. PMID- 19125025 TI - Effects of hearing loss and spectral shaping on identification and neural response patterns of stop-consonant stimuli in young adults. AB - OBJECTIVES: The primary purpose of this study was to more clearly define the effects of hearing loss, separate from age, on perception, and neural response patterns of dynamic spectral cues. To do this, the study was designed to determine whether (1) hearing loss affects the neural representation and/or categorical perception of stop-consonant stimuli among young adults and (2) spectrally shaped amplification aimed at increasing the audibility of the F2 formant transition cue reduces any effects of hearing loss. It was predicted that (1) young adults with hearing loss would differ from young adults with normal hearing in their behavioral and neural responses to stop-consonant stimuli and (2) enhancing the audibility of the F2 formant transition cue relative to the rest of the stimulus would not overcome the effects of hearing loss on behavioral performance or neural response patterns. DESIGN: Behavioral identification and neural response patterns of stop-consonant stimuli varying along the /b-d-g/ place-of-articulation continuum were measured from seven young adults with mild to-moderate hearing impairment (mean age = 21.4 yr) and compared with responses from 11 young adults with normal hearing (mean age = 27 yr). Psychometric functions and N1-P2 cortical-evoked responses were evoked by consonant-vowel (CV) stimuli without (unshaped) and with (shaped) frequency-dependent amplification that enhanced F2 relative to the rest of the stimulus. RESULTS: Behavioral identification and neural response patterns of stop-consonant CVs differed between the two groups. Specifically, to the unshaped stimuli, listeners with hearing loss tended to make low-frequency judgments more often (more /b/, fewer /g/) than listeners with normal hearing when categorizing along the /b-d-g/ continuum. Additionally, N1 amplitudes were larger and P2 latencies were longer to all phonemes in young adults with hearing impairment versus normal hearing. Enhancing the audibility of the F2 transition cue with spectrally shaped amplification did not alter the neural representation of the stop-consonant CVs in the young listeners with hearing loss. It did modify categorical perception such that listeners with hearing loss tended to make high-frequency judgments more often (more /g/, fewer /b/). However, shaping the stimuli did not make their psychometric functions more like those of the normal controls. Instead, young adults with hearing loss went from one extreme (low-frequency judgments with unshaped stimuli) to the other (high-frequency judgments with shaped stimuli), whereas judgments from the normal controls were more balanced. CONCLUSIONS: Hearing loss, separate from aging, seems to negatively impact identification and neural representation of time-varying spectral cues like the F2 formant transition. Enhancing the audibility of the F2 formant transition cue relative to the rest of the stimulus does not overcome the effects of hearing loss on behavioral performance or neural response patterns in young adults. Thus, the deleterious effects of hearing loss on stop-consonant perception along the place of-articulation continuum may not only be due solely to decreased audibility but also due to improper coding by residual neurons, resulting in distortion of the time-varying spectral cue. This may explain, in part, why amplification cannot completely compensate for the effects of sensorineural hearing loss. PMID- 19125026 TI - Frequency tuning curves derived from auditory steady state evoked potentials: a proof-of-concept study. AB - OBJECTIVES: Assess the feasibility of drawing tuning curves from the masking function of steady state potentials. Develop a noninvasive tool for research applications on cochlear frequency selectivity in sedated animals. Obtain pilot human data validating auditory steady state evoked potential-derived (ASSEP) tuning curves against psychophysical data. DESIGN: ASSEP tuning curves were drawn in 10 Beagle puppies and six human adults using amplitude-modulated probes. Two probe frequencies (1 and 2 kHz) were used in dogs and only one (2 kHz) in humans. The modulation rates of the two probes were set to 81 and 88 Hz, respectively. Psychophysical tuning curves were obtained in 12 normal human subjects using the same maskers and either a pure-tone or an amplitude-modulated probe to verify if the latter had a specific effect on tuning curve parameters. Six of these 12 subjects participated in the electrophysiologic measurements. For each tuning curve, the intensity of the narrowband masker required just to mask the fixed probe was plotted for different masker center frequencies. Masker center frequencies extended to about half an octave above and an octave below the probe frequencies in 100-Hz steps. Tuning curve width (Q10 dB values), high- and low frequency slopes (in dB/octave) and the masker frequency yielding the lowest masking threshold (maximal masker frequency) were computed. Canine Q10 dB values obtained were compared with those published for several species with other techniques. For humans, ASSEP and psychophysical tuning curves were directly compared in the same subjects and with published data. RESULTS: In dogs, the ASSEP method yielded reproducible tuning curves with qualitative and quantitative parameters similar to other physiologic measures of tuning obtained in various animals. Q10 dB values were greater at 2 than at 1 kHz, reflecting the well-known correlation between sharpness of tuning and central frequency. In humans, ASSEP Q10 dB values were slightly smaller than the psychophysical ones, but were greater by a factor of 2 than those obtained with previously published electrophysiologic procedures. In both species, detuning-a shift of the tip of the curve away from the probe frequency-was frequently observed as upward shifts with a maximal value of 200 Hz. Human psychophysical tuning curves also showed a certain amount of upward detuning. The intraindividual comparison of the two types of probes performed on human subjects with the psychophysical method did not indicate a specific effect of the amplitude-modulated probe on the curve parameters. Neither did the intraindividual comparisons indicate that an amplitude-modulated probe per se promoted detuning. Detuning has been observed with several other techniques and is usually attributed to nonlinear interactions between masker and probe in simultaneous masking. CONCLUSIONS: The results demonstrate the feasibility of measuring realistic ASSEP tuning curves in sedated dogs and in sleeping human adults. The ASSEP tuning curves exhibit a series of classical features similar to those obtained with time-honored methods. These results pave the way for the development of a noninvasive electrophysiologic method for tuning curve recording and its applications in noncooperative experimental animals or clinical subjects. PMID- 19125027 TI - The derivation of optimum criteria for use in the monothermal caloric screening test. AB - OBJECTIVE: The aim of this study was to determine whether it is possible to predict a normal outcome of the bithermal caloric test by testing at a single temperature and if so, what criteria are most appropriate to use. DESIGN: A total of 490 patients were considered candidates for the bithermal test and 414 completed the four necessary components, their nystagmus being measured using videonystagmography. RESULTS: Clinical decision analysis revealed that the cool monothermal test does not provide an adequate combination of sensitivity and specificity for us to recommend its clinical use. However, the warm monothermal test offers a sensitivity of 95% with 29% of patients with normal bithermal results having to undergo the bithermal test (specificity = 71%) if a combination of three criteria are used: a normal bithermal caloric test outcome can be anticipated and testing curtailed after the first temperature if (a) the warm monothermal caloric asymmetry (MCA) is <15% and (b) the two warm results are each >8 degrees per sec ( degrees /sec), and (c) any spontaneous nystagmus is <4 degrees /sec. CONCLUSIONS: When appropriate criteria are used, the warm monothermal caloric test offers a performance that is acceptable for routine clinical use, sparing a considerable proportion of patients from unnecessary tests at the cool temperature. We believe that the warm/cool monothermal test difference is probably a consequence of the interrelationship between canal paresis and directional preponderance. PMID- 19125028 TI - Widespread auditory deficits in tune deafness. AB - OBJECTIVE: The goal of this study was to investigate auditory function in individuals with deficits in musical pitch perception. We hypothesized that such individuals have deficits in nonspeech areas of auditory processing. DESIGN: We screened 865 randomly selected individuals to identify those who scored poorly on the Distorted Tunes test (DTT), a measure of musical pitch recognition ability. Those who scored poorly were given a comprehensive audiologic examination, and those with hearing loss or other confounding audiologic factors were excluded from further testing. Thirty-five individuals with tune deafness constituted the experimental group. Thirty-four individuals with normal hearing and normal DTT scores, matched for age, gender, handedness, and education, and without overt or reported psychiatric disorders made up the normal control group. Individual and group performance for pure-tone frequency discrimination at 1000 Hz was determined by measuring the difference limen for frequency (DLF). Auditory processing abilities were assessed using tests of pitch pattern recognition, duration pattern recognition, and auditory gap detection. In addition, we evaluated both attention and short- and long-term memory as variables that might influence performance on our experimental measures. Differences between groups were evaluated statistically using Wilcoxon nonparametric tests and t-tests as appropriate. RESULTS: The DLF at 1000 Hz in the group with tune deafness was significantly larger than that of the normal control group. However, approximately one-third of participants with tune deafness had DLFs within the range of performance observed in the control group. Many individuals with tune deafness also displayed a high degree of variability in their intertrial frequency discrimination performance that could not be explained by deficits in memory or attention. Pitch and duration pattern discrimination and auditory gap detection ability were significantly poorer in the group with tune deafness than the normal control group. Approximately one-third of our participants with tune deafness displayed evidence of attention deficit with hyperactivity disorder on the Test of Variables of Attention. Test of Variables of Attention scores were significantly correlated with gap-detection scores, but not significantly correlated with any of the other experimental measures, including the DTT, DLF, and auditory pattern discrimination tests. Short- and long-term memory was not significantly related to any of the experimental measures. CONCLUSIONS: Individuals with tune deafness identified by the DTT have poor performance on many tests of auditory function. These include pure-tone frequency discrimination, pitch and duration pattern discrimination, and temporal resolution. Overall, reduction in performance does not seem to derive from deficits in memory or attention. However, because of the prevalence of attention deficit with hyperactivity disorder in those with tune deafness, this variable should be considered as a potentially confounding factor in future studies of tune deafness and its characteristics. Pure-tone frequency discrimination varied widely in individuals with tune deafness, and the high degree of intertrial variability suggests that frequency discrimination may be unstable in tune-deaf individuals. PMID- 19125029 TI - Smoke alarms for sleeping adults who are hard-of-hearing: comparison of auditory, visual, and tactile signals. AB - OBJECTIVES: People who are hard-of-hearing may rely on auditory, visual, or tactile alarms in a fire emergency, and US standards require strobe lights in hotel bedrooms to provide emergency notification for people with hearing loss. This is the first study to compare the waking effectiveness of a variety of auditory (beeps), tactile (bed and pillow shakers), and visual (strobe lights) signals at a range of intensities. DESIGN: Three auditory signals, a bed shaker, a pillow shaker, and strobe lights were presented to 38 adults (aged 18 to 80 yr) with mild to moderately severe hearing loss of 25 to 70 dB (in both ears), during slow-wave sleep (deep sleep). Two of the auditory signals were selected on the basis that they had the lowest auditory thresholds when awake (from a range of eight signals). The third auditory signal was the current 3100-Hz smoke alarm. All auditory signals were tested below, at, and above the decibel level prescribed by the applicable standard for bedrooms (75 dBA). In the case of bed and pillow shakers intensities below, at, and above the level as purchased were tested. For strobe lights three levels were used, all of which were above the applicable standard. The intensity level at which participants awoke was identified by electroencephalograph monitoring. RESULTS: The most effective signal was a 520-Hz square wave auditory signal, waking 92% at 75 dBA, compared with 56% waking to the 75 dBA high-pitched alarm. Bed and pillow shakers awoke 80 to 84% at the intensity level as purchased. The strobe lights awoke only 27% at an intensity above the US standard. Nonparametric analyses confirmed that the 520 Hz square wave signal was significantly more effective than the current smoke alarm and the strobe lights in waking this population. CONCLUSIONS: A low frequency square wave signal has now been found to be significantly more effective than all tested alternatives in a number of populations (hard-of hearing, children, older adults, young adults, alcohol impaired) and should be adopted across the whole population as the normal smoke alarm signal. Strobe lights, even at high intensities, are ineffective in reliably waking people with mild to moderate hearing loss. PMID- 19125030 TI - Middle ear application of a sodium hyaluronate gel loaded with neomycin in a Guinea pig model. AB - OBJECTIVE: Establishing methods for topical administration of drugs to the inner ear have great clinical relevance and potential even in a relatively short perspective. To evaluate the efficacy of sodium hyaluronate (HYA) as a vehicle for drugs that could be used for treatment of inner ear disorders. METHODS: The cochlear hair cell loss and round window membrane (RWM) morphology were investigated after topical application of neomycin and HYA into the middle ear. Sixty-five albino guinea pigs were used and divided into groups depending on the type of the treatment. Neomycin was chosen as tracer for drug release and pharmacodynamic effect. HYA loaded with 3 different concentrations of neomycin was injected to the middle ear cavity of guinea pigs. Phalloidin stained surface preparations of the organ of Corti were used to estimate hair cell loss induced by neomycin. The thickness of the midportion of the RWM was measured and compared with that of controls using light and electron microscopy. All animal procedures were pe rformed in accordance with the ethical standards of Karolinska Institutet. RESULT: Neomycin induced a considerable hair cell loss in guinea pigs receiving a middle ear injection of HYA loaded with the drug, demonstrating that neomycin was released from the gel and delivered to the inner ear. The resulting hair cell loss showed a clear dose-dependence. Only small differences in hair cell loss were noted between animals receiving neomycin solution and animals exposed to neomycin in HYA suggesting that the vehicle neither facilitated nor hindered drug transport between the middle ear cavity and the inner ear. One week after topical application, the thickness of the RWM had increased and was dependent upon the concentration of neomycin administered to the middle ear. At 4 weeks the thickness of the RWM had returned to normal. CONCLUSION: HYA is a safe vehicle for drugs aimed to pass into the inner ear through the RWM. Neomycin was released from HYA and transported into the inner ear as evidenced by hair cell loss. PMID- 19125031 TI - Development of a mandarin monosyllable recognition test. AB - OBJECTIVE: This study aimed to develop a set of 50- and 25-item word lists that exhibited familiarity, homogeneity, and phonemic balance for a Mandarin monosyllable recognition test (MMRT). DESIGN: To achieve the design goal of high subject familiarity with the test material, we selected the 700 most frequently occurring monosyllables to be the test material. The homogeneity of the test material was achieved by evaluating five psychometric characteristics of these 700 monosyllables to obtain 348 homogeneous monosyllables with similar psychometric functions for constructing the word lists. The phonemic balance of the 50-item word lists was achieved by deriving the desired numbers of initials, finals, and tones in these lists according to their occurrence frequencies in 4733 monosyllabic words. The phonemic balance of the 25-item word lists was achieved by equally dividing the desired numbers of initials, finals, and tones in the 50-item word lists into two groups, called half-A and half-B lists. Three half-A lists and three half-B lists were constructed from the 348 homogeneous monosyllables, and they could be paired to form nine 50-item word lists. Accordingly, all of the MMRT word lists are familiar, homogeneous, and phonemically balanced. RESULTS: The homogeneity of the MMRT word lists was examined by evaluating the interlist equivalence, interitem variability, and intersubject variability. A chi test was used to evaluate the interlist equivalence among the MMRT word lists; that is, to determine whether the values of percent correct recognition were distributed identically at 12 different presentation levels among the six 25-item word lists and nine 50-item word lists. No statistically significant differences were found among the word lists: chi(55) = 12.004 and p = 1.000 for the six 25-item word lists, chi(88) = 7.695 and p = 1.000 for the nine 50-item word lists, and chi(154) = 19.699 and p = 1.000 for the six 25-item word lists plus nine 50-item word lists. The chi tests indicated that the six 25-item word lists and nine 50-item word lists exhibited interlist equivalence. The mean interitem variability (2.0 dB) and mean intersubject variability (2.3 dB) are lower for the MMRT word lists than for previously reported word recognition tests. CONCLUSION: This study applied an innovative approach to design MMRT word lists from familiar and homogeneous monosyllables, in which the familiarity, homogeneity, and phonemic balance of the six 25-item and nine 50-item word lists were strictly controlled. These word lists exhibit interlist equivalence with respect to their psychometric functions and five psychometric characteristics; moreover, their interitem and intersubject variability are lower than those of previously reported lists. Future clinical experiments should examine whether such a design approach can improve the reliability and diagnostic sensitivity of word recognition tests for hearing impaired listeners. PMID- 19125032 TI - Multiple auditory steady state responses (80-101 Hz): effects of ear, gender, handedness, intensity and modulation rate. AB - OBJECTIVE: To evaluate how the amplitudes and latencies of auditory steady state responses (ASSRs) to multiple stimuli presented at rates between 80 and 101 Hz vary with the ear of stimulation, the handedness or gender of a subject, and the rate and intensity of the stimuli. DESIGN: ASSRs were recorded in a group of 56 young adults (27 females, 13 left handed) using several stimulus conditions. In the two main conditions, four sinusoidally amplitude-modulated tones (each uniquely modulated using rates between 80 and 105 Hz) with carrier frequencies of 500, 1000, 2000, and 4000 Hz, were presented concurrently to each ear (eight total). In the first condition the modulation rates for the left ear were slower than those for the right and in the second condition this relationship was reversed. Other conditions evaluated the responses to single stimuli, to multiple stimuli presented in one ear only and to multiple stimuli presented dichotically (four in each ear) with rates that decreased rather than increased with increasing carrier frequency. Stimuli were presented at an intensity of 73 dB SPL except in two conditions wherein the intensity was 53 dB SPL. RESULTS: At 73 dB SPL, multiple-stimulus ASSRs were significantly reduced (monotic or dichotic) compared with single-stimulus ASSRs, especially at 1000 and 2000 Hz. There were significant differences between monotic and dichotic stimulation. When the stimuli were presented dichotically, the amplitude of the response varied with the relative rates of modulation for the stimuli presented in each ear. ASSRs were larger in the ear with the higher rate when the carrier frequencies were 500 and 1000 Hz and when the modulation rates were <90 Hz. There were no consistent effects of gender or ear of stimulation. There were also no significant effects of handedness. CONCLUSIONS: Presenting multiple stimuli at 73 dB SPL in the same ear decreases the amplitude of the ASSR compared with when the stimuli are presented singly. This is caused by the masking effect of low on higher carrier frequencies and some other inhibitory effect of high on lower frequencies. Dichotic stimulation can increase the amplitude of the response to stimuli modulated more rapidly (and concomitantly decrease the responses to the stimuli modulated more slowly). This effect occurs only for carrier frequencies <2000 Hz and for modulation frequencies <90 Hz. Dichotic stimulation also causes a small but highly significant decrease in the latency of the response compared with monotic stimulation. PMID- 19125033 TI - Correlation between audiovestibular function tests and hearing outcomes in severe to profound sudden sensorineural hearing loss. AB - OBJECTIVE: This study investigated whether audiovestibular function tests, namely auditory brain stem response (ABR) and vestibular-evoked myogenic potential (VEMP) tests were correlated to hearing outcomes after controlling the effects of other potential confounding factors in severe to profound sudden sensorineural hearing loss (SSHL). DESIGN: Eighty-eight patients with severe to profound SSHL were enrolled in this study. Pretreatment hearing levels, results of audiovestibular function tests, and final hearing outcomes were recorded from retrospective chart reviews. Other factors, including age, gender, delay of treatment, vertigo, diabetes mellitus, and hypertension, were collected as well. Comparative analysis between multiple variables and hearing outcomes was conducted using the cumulative logits model in overall subjects. Further, multivariate analysis of prognostic factors was conducted in the stratified groups of severe (70 dB HL 90 dB HL) SSHL. RESULTS: Multivariate analysis showed that pretreatment hearing levels, presence of vertigo, and results of ABR and VEMP testing were significant outcome predictors in the overall subjects. Stratification analysis demonstrated that both the presence of ABR and VEMP waveforms were significantly correlated with better hearing outcomes in the group of severe SSHL [ABR: adjusted odds ratio (aOR) = 14.7, 95% confidence interval (CI) = 1.78 to 122, p = 0.01; VEMP: aOR = 5.91, 95% CI = 1.18 to 29.5, p = 0.03], whereas the presence of vertigo was the only significant negative prognostic factor in the group of profound SSHL (aOR = 0.24, 95% CI = 0.06 to 0.95, p = 0.04). Other variables, including age, gender, diabetes mellitus, hypertension, and delay of treatment, were not significantly related to hearing outcomes in both groups (p > 0.05). A predictive hearing recovery table with the combined ABR and VEMP results was proposed for the group of severe SSHL. CONCLUSIONS: ABR and VEMP tests should be included in the battery of neurootological examinations in patients with severe SSHL because the presence of both waveforms might indicate favorable hearing outcomes. The presence of vertigo might portend a worse prognosis in patients with profound SSHL. A presumed table in predicting hearing outcomes was suggested for patients with severe SSHL. PMID- 19125034 TI - Relation of electrically evoked compound action potential thresholds to behavioral T- and C-levels in children with cochlear implants. AB - OBJECTIVE: This study examined the relation of electrically evoked compound action potential thresholds obtained using neural response telemetry (NRT) to T- and C-levels in children's speech processor programs optimized for recognition of very soft to loud sounds while ensuring tolerance of very loud sounds. DESIGN: Forty-one children (age 2 to 14 yr) with stable electrical hearing participated. All children were Nucleus 24 System recipients and attended one of three auditory oral schools that have on-site pediatric audiologists experienced at cochlear implant programming. Speech processor MAPs were created and adjusted over a period of months until aided warble-tone thresholds were between 10 and 30 dB HL at octave frequencies between 250 and 4000 Hz, and understanding of speech was maximized for many listening situations. At least 1 yr postactivation, visual (vNRT) and predicted (tNRT) thresholds were obtained on 9 to 11 electrodes and compared to each child's T- and C-level values on these electrodes in their MAPs. Test-retest stability of NRT thresholds was compared for two test sessions 1 mo apart. RESULTS: NRT-based evoked compound action potential thresholds could be obtained from 36 of the 41 children. vNRT and tNRT test-retest reliability was high; average correlation coefficients (r) across subjects were 0.90 (range: 0.64 to 0.99) and 0.88 (range: 0.31 to 1.00), respectively. Group average correlation coefficients between vNRT and T-level, vNRT and C-level, tNRT and T-level, and tNRT and C-level were low (0.18, 0.21, 0.24, and 0.26, respectively). Group mean tNRT thresholds were four current levels lower than the group mean vNRT thresholds. Subsequent analysis was performed with the vNRT thresholds because the range of test-retest correlation coefficients for individual subjects was narrower than with tNRT. Hierarchical linear modeling was used to determine if vNRT could be used to predict T- and C-levels. This analysis indicated a significant average relation between vNRT and T-levels and between vNRT and C levels, but significant heterogeneity in the individual-level estimates of those relations. In other words, subjects varied significantly in the size of the relation between their individual vNRT values and both T- and C-levels. Attempts to account for that heterogeneity did not identify any subject characteristics that were significantly related to the individual-level parameters. CONCLUSIONS: The position of the group average vNRT and tNRT thresholds in the upper half of the dynamic range between Ts and Cs agrees with previous studies. The fact that the profile of vNRT thresholds did not parallel the profiles of Ts and Cs across electrodes for most children suggests that simply shifting the NRT profile to select T- and C-levels in initial MAPs is likely to result in a loudness imbalance for certain speech frequencies and/or tolerance issues for many children. This was verified by the hierarchical linear modeling analysis, which showed substantial and significant heterogeneity in the relations between vNRT and T-levels and between vNRT and C-levels. In summary, vNRT is not related to T- or C-levels in a simple and uniform way that would allow it to guide MAP fine tuning with any precision. Consequently, it is recommended that MAP fine tuning be based on the child's behavioral responses on individual electrodes. PMID- 19125035 TI - Receptive vocabulary development in deaf children with cochlear implants: achievement in an intensive auditory-oral educational setting. AB - OBJECTIVES: Deaf children with cochlear implants are at a disadvantage in learning vocabulary when compared with hearing peers. Past research has reported that children with implants have lower receptive vocabulary scores and less growth over time than hearing children. Research findings are mixed as to the effects of age at implantation on vocabulary skills and development. One goal of the current study is to determine how children with cochlear implants educated in an auditory-oral environment compared with their hearing peers on a receptive vocabulary measure in overall achievement and growth rates. This study will also investigate the effects of age at implant on vocabulary abilities and growth rates. We expect that the children with implants will have smaller vocabularies than their hearing peers but will achieve similar rates of growth as their implant experience increases. We also expect that children who receive their implants at young ages will have better overall vocabulary and higher growth rates than older-at-implant children. DESIGN: Repeated assessments using the Peabody Picture Vocabulary Test were given to 65 deaf children with cochlear implants who used oral communication, who were implanted under the age of 5 yr, and who attended an intensive auditory-oral education program. Multilevel modeling was used to describe overall abilities and rates of receptive vocabulary growth over time. RESULTS: On average, the deaf children with cochlear implants had lower vocabulary scores than their hearing peers. However, the deaf children demonstrated substantial vocabulary growth, making more than 1 yr's worth of progress in a year. This finding contrasts with those of previous studies of children with implants, which found lower growth rates. A negative quadratic trend indicated that growth decelerated with time. Age at implantation significantly affected linear and quadratic growth. Younger-at-implant children had steeper growth rates but more tapering off with time than children implanted later in life. CONCLUSIONS: Growth curves indicate that children who are implanted by the age of 2 yr can achieve receptive vocabulary skills within the average range for hearing children. PMID- 19125036 TI - Perception of envelope-enhanced speech in the presence of noise by individuals with auditory neuropathy. AB - OBJECTIVE: To investigate the effect of background noise on perception of unprocessed and envelope-enhanced speech in individuals with auditory neuropathy (AN). DESIGN: Open-set speech identification abilities were assessed for unprocessed and envelope-enhanced speech in quiet and at three signal to noise ratios (10, 5, and 0 dB signal to noise ratio) in 15 individuals with AN. RESULTS: Results revealed that speech identification ability was affected more in the presence of noise than in quiet. The participants were classified into two groups, Good Performers and Poor Performers, based on their speech identification scores in quiet. It was observed that the reduction in mean speech identification scores in noise was less for Good Performers when compared with that of Poor Performers. Envelope enhancement of the signal improved speech identification scores in both the groups. The analysis of results also showed that Good Performers showed a significant improvement in all the conditions, whereas Poor Performers showed improvement only in quiet and +10 dB signal to noise ratio. CONCLUSIONS: For individuals with AN, listening in background noise is more difficult than in quiet condition. Envelope enhancement of speech signal improves speech identification scores in quiet and in background noise for individuals with AN. However, the benefit obtained varies depending on the speech identification ability in quiet. PMID- 19125037 TI - When an air-bone gap is not a sign of a middle ear conductive hearing loss. PMID- 19125040 TI - Amoebicidal activities of alexidine against 3 pathogenic strains of acanthamoeba. AB - OBJECTIVES: Effective pharmacotherapy for Acanthamoeba keratitis has been hampered because of the marked resistance of various stains to a variety of antimicrobial agents. In view of the fact that topical Brolene (propamidine isethionate) and neosporin are currently considered to be the first-line medical treatment of choice in Europe, we sought to determine whether Alexidine is equally effective, because the latter drug is more readily available in the United States. METHODS: Trophozoites and cysts from 3 pathogenic corneal isolates (A. castellanii, A. polyphaga, and A. rhysodes) were incubated in peptone-yeast extract-glucose medium containing different concentrations of Alexidine for 24 hr. The number of trophozoites was counted by hemocytometer. The cysts were plated in to nonnutrient agar plates precoated with Escherichia coli and observed for viability or excystment over a period of 2 weeks. The capacity of different concentrations of Alexidine to induce cytolysis of corneal epithelial cells was tested in vitro. Chinese hamster corneas were treated with 5 microL of Alexidine topically, every hour; 6 times a day and the corneas were stained with fluorescein to asses the epithelial defects in vivo. RESULTS: Alexidine was effective in killing the trophozoites at a concentration of 10 microg/mL. However, a higher concentration of Alexidine (100 microg/mL) is required to kill Acanthamoeba cysts and the cytotoxic activities of Alexidine are comparable with chlorhexidine. We have also demonstrated that both Alexidine and chlorhexidine at 100 microg/mL induced significant cytopathic effect on the corneal epithelial cells in vitro. In vivo results indicate that Alexidine at a concentration of 100 microg/mL is less toxic than chlorhexidine when applied topically to the Chinese hamster cornea. CONCLUSIONS: Our study has identified Alexidine as a novel anti Acanthamoeba drug and suggests that Alexidine may be an effective therapeutic option because of its potency and low toxicity to the corneal tissues when applied topically in vivo. PMID- 19125041 TI - The impact of test medium on use of visual analogue scales. AB - OBJECTIVES: Visual analog scales are frequently used as a means of allowing participants to rate symptoms during clinical trials. The accuracy and reproducibility of these scales play an important role in determining the experimental value of the data they provide. This study was initiated to compare the data collected using paper- and computer-based (Tablet PC) analog scales to better understand the variability in data provided by a visual analog scale. METHODS: Thirty participants rated ocular comfort, redness, and clarity of vision (right and left eyes) on a nondemarcated horizontal line on both paper and a Tablet PC. Measurements were taken in the morning between the hours of 8:30 and 10:30 am and again the same day between 2:30 and 4:30 pm. RESULTS: The mean difference between the measures recorded in the morning for the 2 media was 2.6 +/- 0.9 (confidence intervals, 2 standard errors of the differences) units on a 100 unit scale, with the Tablet PC having the higher mean measure. The limits of agreement (2 standard deviations of the differences) was 9.4 units. Comparing the difference of the differences (1.0 +/- 1.3) between the 2 methods of measure (morning vs. afternoon) the visual analog scales on the Tablet PC seemed to have good reproducibility of agreement in comparison with the paper version. CONCLUSIONS: Discrepancy analysis yielded no significant difference and slight bias between paper- and computer-based analog scales. Repeatability of measures using the Tablet PC was also demonstrated. These results suggest that the choice of medium does not significantly influence the outcome for subjective analog scales. PMID- 19125042 TI - Anomalies in the prescribing of soft contact lens power. AB - OBJECTIVES: To determine the proportion of prescribed soft lenses rounded to the nearest half diopter and any variations from country to country and between lens types. METHODS: Marketing data were obtained for soft lenses supplied during a 1 year period for lenses representing each of the following categories: mid-water hydrogel (MWH), silicone hydrogel, daily disposable, and toric silicone hydrogel (TSH). The data were analyzed for several countries/regions. Spherical lenses were analyzed in the range 1.00 to 5.75 D for plus and minus powers, and toric lenses in the range 0.50 to 5.75 D. This ensured a similar number of lenses in full or half diopter powers were compared with quarter and three-quarter diopter powers, and that there was no enforced rounding due to nonavailability of powers. By comparing the proportion of lenses from the 2 power groups, the proportion of lenses rounded to the nearest half diopter was estimated. It was assumed that half the difference between the totals of the 2 power groups represented those lenses dispensed to the nearest half diopter and, therefore, dispensed inaccurately; this was termed the "rounding rate" (RR). RESULTS: The power distribution curve for the sphere powers spiked in half diopter steps, illustrated a bias toward prescribing full and half diopter powers. With all lenses, the RR varied widely between countries. For the MWH, this ranged from 1.7% (Canada) to 11.6% (Iberia). The RRs were 2 to 3 times higher for plus than minus power lenses, however, this also varied by country. Overall, the RRs were lower for the silicone hydrogel and daily disposable contact lenses compared with the MWH, in particular for France and Iberia. The TSH results showed the greatest consistency between countries, with RRs ranging from 3.9% (Germany) to 9.5% (Rest of Europe). Most countries showed similar or lower RRs for TSH compared with MWH although, for some countries (e.g., United Kingdom, Nordic), these were higher. There was less difference in RRs for TSH lenses between plus and minus spheres. CONCLUSION: A surprising proportion of soft lenses are prescribed to the nearest half diopter, although this varies according to lens type. There are also considerable variations between countries, presumably due to differences in training, fitting practices, and supply routes. These findings suggest that there is widespread room for improvement in the prescribing accuracy of soft contact lenses. PMID- 19125043 TI - Unilateral keratoconus: videokeratography and Orbscan study--optical correction. AB - OBJECTIVES: To assess the frequency of unilateral keratoconus (KC) and vision correction with a contact lens (CL) in the KC eye. METHODS: Transverse study in which we revised the clinical records of 385 KC patients. Of these, 35 (9%) showed clinical unilateral KC. Corneal topography was evaluated by a Placido disk and an elevation topographer. We also determined distance and near visual acuity before and after correction with spectacles or CL in KC eye. RESULTS: Mean central corneal curvature (K) of the KC eyes was 51.45 +/- 6.65 D, versus 43.58 +/- 1.71 D for the fellow eye (P<0.001). Mean maximum curvature of the inferior cornea was 54.62 +/- 5.91 D in the KC eye and 44.66 +/- 2.38 in the fellow eye (P<0.001). Mean of the thinnest point pachymetry was 411 +/- 55 microm in the KC eye and 477 +/- 42 microm in the contralateral eye (P<0.0001). In most of the 29 patients in whom CLs were fitted, aspheric rigid gas-permeable CLs were used, corrected visual acuities were significantly better (P<0.0001) than without correction. CONCLUSIONS: All unilateral KC patients showed higher corneal curvature in the affected eye. Corneal thickness was likewise clearly reduced in the KC eye, and reduced with respect to normal values in the contralateral eye. Aspheric rigid gas-permeable CL were the most effective for improving distance and near visual acuity in patients with adequate tolerance. PMID- 19125044 TI - Repeatability of central corneal thickness measures by Orbscan pachymetry for right and left eyes. AB - OBJECTIVES: To assess the repeatability of Orbscan measures on right versus left eyes. METHODS: Three Orbscan measures were taken from the right eye and then the left eye by the same operator on 50 healthy myopic subjects (average right eye = 4.3 DS) aged 19 to 58 years, half of whom were successful long-term soft contact lens (SCL) wearers. The central point thickness measurement was compared for the 3 scans for each eye, the variability calculated based on the coefficient of variation (COV) and also paired comparisons with limits of agreement. No acoustic factor was applied. RESULTS: Single central point thickness values ranged from 0.483 to 0.663 mm, with mean values of 0.579 mm for both eyes, with no statistically significant difference between spectacle wearers and SCL wearers (P>or=0.387). From 3 repeat scans, the net difference was -0.001 mm between third and first scans of right eyes and +0.001 mm for left eyes (P>or=0.5). Overall, the COV was close to 1%, being marginally higher (0.955% and 1.176%) in spectacle wearers than SCL wearers (0.922% and 0.836%; P, NS). However, an overall assessment of the COV data indicates that the values were inversely related to absolute thickness values, being slightly higher for thinner versus thicker corneas (P or =20,000/microL of platelet. CONCLUSIONS: These findings indicate that overall compliance to the Japanese guideline is considered to be relatively good in Japan. PMID- 19125085 TI - Clinical nurse coordinators: a new generation of highly specialized oncology nursing in Jordan. AB - Despite the important role of nurses in a pediatric oncology team, the marginalization of nursing is common, especially in developing countries. At the King Hussein Cancer Center (KHCC) in Jordan, the position of the clinical nurse coordinator (CNC) was created in the pediatric neuro-oncology and ocular oncology services to empower the role of nursing. Our advanced nursing practice is based on the domains of the Strong Model of Advanced Practice, which are direct patient care, support of systems, education, research and publications, and professional leadership. There is strong involvement of the CNC in clinical care, patient education, follow-up, and interdisciplinary collaboration, which has resulted in improved survival, reduced morbidity, and increased compliance of patients and also their successful reintegration into society. The coordination of twinning initiatives and telemedicine activities has helped ensure the quality of treatment and supported research initiatives in the services. Such a positive impact has led to a significant increase in the recruitment of CNCs at KHCC. We propose that the role of nursing be strengthened in developing countries to improve the level of care provided to patients and their families. PMID- 19125084 TI - Low-level infrared laser therapy in chemotherapy-induced oral mucositis: a randomized placebo-controlled trial in children. AB - BACKGROUND: Oral mucositis (OM) is one of the most frequent complications of chemotherapy for which there is no standard therapy; treatment is mostly conservative. This study was conducted to determine whether low-intensity laser therapy (LLLT) can reduce the duration of chemotherapy-induced OM. PROCEDURE: A placebo-controlled randomized trial was carried out using LLLT or placebo (sham treatment). Children and adolescents with cancer receiving chemotherapy or hematopoietic stem-cell transplantation between October 2005 and May 2006 were eligible as soon as they developed OM. Patients received intervention for 5 days. The LLLT group was treated with laser GaAlAs, wavelength (lambda): 830 nm (infrared), power: 100 mW, dose: 4 J/cm, and placebo group underwent sham treatment. The grade of OM was clinically assessed by the National Cancer Institute, Common Toxicity Criteria scale. RESULTS: Twenty-one patients developed OM and were evaluable for analysis; 18 (86%) patients had a diagnosis of leukemia or lymphoma and 3(14%) had solid tumors. The mean age was 8.2 (+/-3.1) years. Nine patients were randomized in the laser group and 12 in the placebo-control group. Once OM was diagnosed, the patients had daily OM grading assessments before laser or sham application and thereafter until complete healing of the lesions. On day 7 after OM diagnosis, 1/9 of patients remained with lesions in laser group and 9/12 of patients in the placebo-control group (P=0.029). In the laser group, the mean of OM duration was 5.8+/-2 days and in the placebo group was 8.9+/-2.4 days (P=0.004). CONCLUSIONS: Our study has shown evidence that laser therapy in addition to oral care can decrease the duration of chemotherapy induced OM. Our results confirm the promising results observed in adult cancer patients and should encourage pediatric oncologists to use laser therapy as first line option in children with chemotherapy-induced OM. PMID- 19125086 TI - Hydroxyurea or chronic exchange transfusions in patients with sickle cell disease: role of transcranial Doppler ultrasound in stroke prophylaxis. AB - Hydroxyurea is increasingly used in patients with sickle cell disease and acts by several underlying mechanisms. Its usage has been reported to eliminate transfusion needs in children with beta-thalassemia major. It has also been used in sickle cell disease patients on stroke prophylaxis with exchange transfusions who develop transfusional iron overload and is now being considered as a possible alternative to chronic transfusions for secondary stroke prophylaxis. Our case demonstrates a pitfall when using hydroxyurea without monitoring intracranial cerebral vessels for vasculopathic changes; presence of which can predict stroke risk and alert the need for chronic blood transfusions as primary stroke prophylaxis. Transcranial Doppler is a crucial investigation that can reveal elevated cerebral arterial flow velocities. PMID- 19125087 TI - Adolescent wilms tumor with intraspinal and bone metastases: a case report and the review of literature. AB - A 14-year-old girl was referred for a large tumor of the left kidney, with intraspinal and vertebral metastases. Left nephrectomy and intraspinal tumor resection were performed. The histology of both tumors was nephroblastoma with no anaplasia and favorable histology, and they were diagnosed as stage IV. The tumor bed and vertebras were irradiated. We started chemotherapy according to the DD-4A regimen of Japanese Wilms' Tumor Study Group. The vertebral metastasis was additionally irradiated. The patient has remained in disease-free remission for 45 months after the surgical resection. Intensive multimodality therapy including DD-4A regimen of National Wilms' Tumor Study can result in long-term disease-free remission. PMID- 19125088 TI - Successful treatment of hodgkin lymphoma in nijmegen breakage syndrome. AB - Nijmegen breakage syndrome (NBS) is a rare DNA repair disorder characterized by microcephaly, immunodeficiency, and predisposition to malignancy. We report on a 5-year-old patient with NBS who presented with nodular sclerosing type of Hodgkin disease stage IVB. Chemotherapy consisting of COPP/ABV regimen with reduction at 75% of full doses was employed. During this treatment, no major toxic or infectious complications were observed. Complete remission was achieved lasting now for 20 months. In DNA repair disorders, prognosis of Hodgkin disease is poor as opposed to excellent overall prognosis in general pediatric population. Better survival may be achieved both with adopted, disease-specific regimens, and individualized approach considering patient's clinical condition. Also, better recognition and treatment of infections during chemotherapy may reduce early deaths in patients with DNA repair disorders. PMID- 19125089 TI - Treatment of refractory Langerhans cell histiocytosis (LCH) with a combination of 2-chlorodeoxyadenosine and cytosine arabinoside. AB - The combination of 2-chlorodeoxyadenosine (2-CDA) and cytosine arabinoside (Ara C) has been shown to be effective in children with refractory Langerhans cell histiocytosis (LCH). We have treated 5 patients with recurrent LCH with 2-CDA/Ara C chemotherapy and closely followed immune and hematopoietic function. These patients display a decline in the absolute CD4, CD8, and natural killer cell number, decrease in the CD4/CD8 ratio. Septic events, including pneumocystis infection were present after most of the treatment courses (15/21). These data suggest that 2-CDA /Ara-C, should be considered in resistant and relapsed pediatric patients with LCH with high-risk multiorgan involvement. Consequent profound prolonged combined immune deficiency and myelosupression should be anticipated. PMID- 19125090 TI - Intrathecal methotrexate-induced acute chorea. AB - Intrathecal methotrexate is the mainstay of central nervous system prophylaxis in acute lymphoblastic leukemia. We describe a patient who developed acute chorea after last dose of intrathecal methotrexate therapy and recovered completely. PMID- 19125091 TI - Congenital stage 1 neuroblastoma evolved into stage 4s. AB - A newborn with a prenatally detected adrenal mass underwent complete resection of a stage 1 favorable histology neuroblastoma (NB) without MYC-N amplification. Two months later, the infant presented with a local recurrence and multiple hepatic metastases. Close follow-up without therapy was adopted for stage 4s NB. Enlarging tumor lesions were seen until the child was 8 months old, followed by later decrease in size. At 36 months of follow-up, the child is alive and disease free. We describe this case of NB and its abnormally short evolution from stage 1 to stage 4s, despite initial surgery. Its spontaneous regression may help us understand the natural history of congenital NB. PMID- 19125092 TI - Primary mixed glioneuronal tumor of the central nervous system in a patient with noonan syndrome: a case report and review of the literature. AB - Noonan syndrome is an autosomal dominant condition with variable phenotypic expression. Although an association between Noonan syndrome and various neoplasms has been identified, a relationship with primary glial or neuronal tumors of the central nervous system (CNS) has not yet been established. We describe the case of a 6-year-old male patient with Noonan syndrome and leptomeningeally disseminated low-grade mixed glioneuronal tumor. After a literature review, this case emerges as the third patient to present with Noonan syndrome and primary CNS glial tumor and the first with mixed glioneuronal tumor, indicating the possible association between these individual entities. PMID- 19125093 TI - Histiocytic sarcoma presenting with chylous ascites in a 7-month-old infant: a case report. AB - We describe in this report what we believe to be the first report of a rare presentation of a very rare tumor, especially in this age group. We highlight the importance of early consideration of malignancy as a cause of chylous ascites in infancy and we discuss different causes of chylous ascites. PMID- 19125094 TI - Assessment of LV diastolic function in patients with beta-thalassemia major with special reference to E/Eann ratio. AB - OBJECTIVE: The present study was undertaken to evaluate left ventricular function in patients with beta-thalassemia major with special reference to pulsed wave tissue Doppler imaging. METHODS: The present study compared 30 diagnosed cases of beta-thalassemia major (mean age: 9.43+/-2.78 y) with 30 age-sex-matched healthy controls (mean age: 8.3+/-2.83 y) for left ventricular function assessment using following parameters: ejection fraction, mitral valve E/A ratio and E deceleration time (Edec), isovolumic relaxation time, and E/Eann ratio. Serum ferritin levels were also measured. RESULTS: There was significant increase in E/Eann (9.46+/-1.5 vs. 6.16+/-2.4, P<0.0001) and significant prolongation of Edec (177.66+/-40.73 vs. 138.5, P<0.001) and isovolumic relaxation time (40.2+/-14.89 vs. 36.67+/-5.12, P<0.05) in cases as compared with controls. However, there was no significant difference in ejection fraction value (65.55+/-8.98 vs. 63.87+/ 16.35) and E/A ratio (2.0386+/-0.73 vs. 2.119+/-0.92). Serum ferritin levels although increased significantly in cases (8370.85+/-2660.35), no correlation could be established between increased serum ferritin and progressive diastolic dysfunction (r=0.148, P=0.258). CONCLUSIONS: Diastolic dysfunction precedes systolic dysfunction in patients with beta-thalassemia major as assessed by Edec and E/Eann ratio. PMID- 19125095 TI - False positivity of FDG-PET during hemophagocytic lymphohistiocytosis in a child with Hodgkin lymphoma in remission. PMID- 19125096 TI - Multifocal osteomyelitis as a presenting manifestation of childhood acute myeloid leukemia. PMID- 19125097 TI - Transient spontaneous remission after tumor lysis syndrome triggered by a severe pulmonary infection in an adolescent boy with acute lymphoblastic leukemia. PMID- 19125098 TI - Poor treatment compliance in children with down syndrome and acute lymphoblastic leukemia. PMID- 19125099 TI - Editor's message: the importance of context and the upper regulatory elements. PMID- 19125100 TI - Fitness test results of Hungarian and international-level soccer referees and assistants. AB - The aim of our research was primarily to compare the conditional abilities of European countries' Federation Internationale de Football Association (FIFA) referees with those of Hungarian national and county referees and, secondly, to investigate whether the stop tests of FIFA and the Union of European Football Associations (UEFA) provide reliable discrimination criteria between the different levels of referees. The main focuses of our research were speed and endurance, which were measured by the stop tests of FIFA and UEFA-namely, 50- and 200-m sprint runs and a 12-minute run, respectively (Cooper test). In our study, the test results of 2459 soccer referees were analyzed; they include the results of the total Hungarian population of referees and the results from 17 different European countries. Referees were classified into 6 different levels according to their qualifications. The results show that in the 12-minute run, FIFA referees covered the longest distance (3043 +/- 127 m), followed by the Hungarian elite referees (2939 +/- 136 m); less distance was covered by county III-level referees (2522 +/- 270 m). Differences between groups in the endurance event were significant (p < 0.05) in all cases. In the sprint runs, there was no difference between FIFA and Hungarian elite referees. In both sprints, FIFA-level and Hungarian elite referees performed the fastest times, and county III-level referees had the slowest times. It can be concluded that the aerobic fitness level demonstrated in the 12-minute run provides the most adequate criteria for discrimination between different competitive levels. Hungarian elite referees, on the basis of their fitness test results, are comparable with international-level referees. The major quality step in a referee's career can be located at the third-division competition level. PMID- 19125101 TI - The effect of resisted sprint training on speed and strength performance in male rugby players. AB - Various studies have demonstrated that resistance sprint (RS) training can produce significant changes in running speed and running kinematics. The longer term training adaptations after RS training remain unclear. The purpose of this study was to investigate whether an RS training intervention would enhance the running speed and dynamic strength measures in male rugby players. Fifteen male rugby players aged 20.5 (+/- 2.8) years who were proficient in resisted sledge training took part in the study. The subjects were randomly assigned to control or RS groups. The RS group performed two sessions per week of RS training for 6 weeks, and the control group did no RS training. Pre- and postintervention tests were carried out for 30-m sprint, drop, squat, and rebound jumps on a force sledge system. A laser measurement device was used to obtain velocities and distance measures during all running trials. The results show a statistically significant decrease in time to 5 m for the 30-m sprint for the RS group (p = 0.02). The squat jump and drop jump variables also showed significant increases in starting strength (p = 0.004) and height jumped (p = 0.018) for the RS group from pre- to post-testing sessions. The results suggest that it may be beneficial to employ an RS training intervention with the aim of increasing initial acceleration from a static start for sprinting. PMID- 19125102 TI - Positive association of neuroligin-4 gene with nonspecific mental retardation in the Qinba Mountains Region of China. AB - OBJECTIVE: Neuroligin-4 is essential for proper brain function. Some studies indicate a close relationship between neuroligin-4 and several human psychiatric conditions. METHODS: The case-control method was used to study the association between nonspecific mental retardation (NSMR) and genetic variants of neuroligin 4 gene (NLGN4). Five single nucleotide polymorphisms (SNPs: rs5916271, rs7049300, rs6638575, rs3810686, and rs1882260) were genotyped by PCR-RFLP/SSCP method in the NLGN4. RESULTS: Individual SNP analysis shows significant differences at SNPs rs3810686 and rs1882260 for allele frequency when NSMR cases and controls [odds ratio (OR)=1.589, 95% confidence interval (CI)=1.035-2.438, chi2=4.53, df=1, P=0.033; OR=2.050, 95% CI=1.211-3.470, chi2=7.38, df=1, P=0.007, respectively] were compared. Further haplotype analysis indicates that there are two haplotype sets, rs3810686-rs1882260 and rs6638575-rs3810686-rs1882260, which show statistical differences between NSMR cases and controls (chi2=6.79, df=2, global P=0.034; chi2=9.29, df=2, global P=0.0096, respectively). CONCLUSION: The results suggest a positive association between the genetic variants of the NLGN4 and NSMR in the Chinese children from Qinba Mountains Region. PMID- 19125103 TI - Association analysis of the glutamic acid decarboxylase 2 and the glutamine synthetase genes (GAD2, GLUL) with schizophrenia. AB - OBJECTIVE: As dysfunction of glutamatergic neurotransmission is one of the plausible hypotheses for the pathogenesis of schizophrenia, genes involved in the glutamate neurotransmitter system are candidates for schizophrenia susceptibility. The aim of this study is to clarify the contribution of two genes encoding glutamate metabolic enzymes: the glutamic acid decarboxylase 2 gene (GAD2) and the glutamine synthetase gene (GLUL), in schizophrenia. METHODS: We genotyped 300 Japanese schizophrenia patients and 300 healthy controls for 14 single nucleotide polymorphisms (SNPs) in GAD2 (approximately 91 kb in size) and six SNPs in GLUL (approximately 14 kb in size). We examined 'single-point' association as well as pairwise haplotype association for all SNPs with schizophrenia. RESULTS: We observed no significant 'single-point' associations with the disease in any of the 20 SNPs after correction for multiple testing using False Discovery Rate. We also observed no significant haplotype associations with False Discovery Rate. Furthermore, we analyzed gene-gene interactions, including six glutamate receptor genes we have reported previously in the association studies of GRIA4, GRIN2D, GRIK3, GRIK4, GRIK5, and GRM3, using the multifactor dimensionality reduction method. The best interaction model, however, did not show the statistical significance. CONCLUSION: These results suggest that GAD2 and GLUL do not play a major role in schizophrenia pathogenesis and there is no gene-gene interaction between the eight genes in the Japanese population. PMID- 19125104 TI - Trajectories of depressive symptoms, dopamine D2 and D4 receptors, family socioeconomic status and social support in adolescence and young adulthood. AB - OBJECTIVES: The purpose of this study is two-fold. First, we tested the association between dopamine D2 and D4 receptors and a trajectory of depressive symptoms in adolescence and young adulthood. Second, we reestimated the association between the dopamine receptors and depression after taking into account the effects of socioeconomic disparity and child-parent ties and social support. METHODS: The study uses the DNA sample of approximately 2500 individuals in the National Longitudinal Study of Adolescent Health (Add Health). Each individual was measured three times in 1994, 1996, and 2002. RESULTS: This study has yielded robust associations of the DRD2 and DRD4 variants with depressive symptoms among male adolescents and young adults. The DRD2*304/178 genotype is associated with a level of depressive symptoms 0.04-0.07 points (3-5% of the mean) higher than the DRD2*178/178 genotype. Relative to the other more common DRD4 variants, the DRD4*379/379 genotype raises the level of depression by about 0.25 points (about 17% of the mean). These findings hold after adjusting for the effects of socioeconomic status (family structure, parental education, family income, mother's employment status, and whether attending public school) and child-parent ties/social support (conflict with parent(s), closeness to parent(s), parental availability, and social support). Although the gene-sex interaction is clearly present, the tests of gene-lifecourse interaction did not yield any significant results. CONCLUSION: Our findings emphasize the importance of joint influences of genetic propensities and social environment on depressive symptoms. PMID- 19125105 TI - Mental retardation and Xq12-Xq23: candidate loci for nonspecific mental retardation in the male population of the QinBa region. AB - OBJECTIVES: The higher prevalence of nonspecific mental retardation (NSMR) presents an important socioeconomic and medical issue for families and the whole QinBa region in China. The obvious family aggregation and high heritability indicated that genetic causes play a role in the NSMR population in QinBa. This study discusses the relationship between Xq12-Xq23 region and NSMR in the QinBa area. METHOD: We chose six short tandem repeats--DXS7132, DXS6979, DXS1191, DXS1230, DXS1072, and DXS6804, located in Xq12-Xq23--and analyzed the distribution difference of their alleles between the NSMR and control boys. RESULTS: A significant allele distribution difference was found between NSMR and control boys (all P<0.05) for DXS7132, DXS1191, DXS1230, DXS1072, and DXS6804 but not for DXS6979. CONCLUSION: Our results suggest that Xq12-Xq23 may be the candidate region where there are one or more loci, linked to NSMR in the QinBa region. Further study needs to be carried out for locating the gene responsible for NSMR in this region and a larger sample size and more genetic markers are needed. PMID- 19125106 TI - Role of shared genetic and environmental factors in symptoms of depression and body composition. AB - BACKGROUND: Both obesity and lean mass have been correlated with symptoms of depression. OBJECTIVE: To investigate the contribution of genetic and environmental factors in the co-occurrence of obesity and lean mass with symptoms of depression. METHODS: Individuals were 2383 participants of the Erasmus Rucphen Family study. Symptoms of depression were assessed using the Center for Epidemiologic Studies Depression Scale and the Hospital Anxiety and Depression Scale. Anthropometric and dual-energy X-ray absorptiometry total body scans were obtained for the assessment of body composition. The role of shared genetic and shared environmental factors was quantified by estimating genetic and environmental correlations between symptoms of depression and measures of body composition. RESULTS: Phenotypic correlations between body composition and symptoms of depression ranged from -0.08 to 0.08. Heritability estimates for body composition ranged from 0.40 to 0.46 (P<0.001) in women and from 0.35 to 0.51 (P<0.001) in men, and heritability estimates for depression scores were higher in women (0.34 and 0.37) than in men (0.13 and 0.21). No consistent genetic correlations between measures of body composition and symptoms of depression were found. We did find a significant consistent environmental correlation between depression scores and lean mass index (environmental correlation=-0.23 for Center for Epidemiologic Studies Depression Scale and -0.31 for Hospital Anxiety and Depression Scale). CONCLUSION: In our study, there is no evidence that the co occurrence of symptoms of depression and body composition result from a common genetic pathway. PMID- 19125107 TI - Effect of the 5-HTTLPR polymorphism in the serotonin transporter gene on major depressive disorder and related comorbid disorders. AB - OBJECTIVES: The serotonin transporter gene (5-HTT) has been proposed as a candidate gene for major depressive disorder (MDD). Association studies, however, have revealed inconsistent results. This could be because of the phenotypic heterogeneity of MDD, as it often presents with comorbid disorders such as generalized anxiety disorder (GAD), alcohol-related disorders, and dysthymia. METHODS: In this exploratory study, we performed regression analyses with generalized estimating equations in patients with familial MDD (n=233) in order to explore whether a polymorphism in the serotonin transporter gene (5-HTTLPR) is differentially associated with MDD and a comorbid disorder compared with MDD without that particular comorbidity. As in general, GAD is more common in females and alcohol-related disorders are more common in males, the analyses were stratified for sex. RESULTS: Comorbid dysthymia was less common in s-allele carriers with MDD (P<0.05) than in patients homozygous for the long allele. In the sex-specific analyses, an association between the 5-HTTLPR and comorbid alcohol use disorders was observed in females, with s-carriers reporting significantly less alcohol use disorders. The relationship with comorbid GAD differed by sex with male s-carriers reporting more comorbid GAD than female s carriers. CONCLUSION: The effect of the 5-HTTLPR polymorphism on MDD is co dependent on the presence of comorbid disorders and sex. In this study, the s allele of the 5-HTTLPR polymorphism was associated with significantly lower rates of particular lifetime comorbid disorders. Therefore, the presence of comorbid psychiatric disorders should be taken into account to clarify the association of the 5-HTTLPR polymorphism with MDD phenotypes. PMID- 19125108 TI - Neuropsychological performance as endophenotypes in extended schizophrenia families from the Central Valley of Costa Rica. AB - OBJECTIVE: The understanding of complex heritable psychiatric disorders such as schizophrenia could be clarified by examining endophenotypes within genetically isolated populations, such as the one found in the Central Valley of Costa Rica. The reduction of familial variability within a sample could allow the relationship between the cognitive and symptomatic manifestations of the illness and the genetic underpinnings to become more observable. This study investigates the neuropsychological test performances of 41 family members from four extended multiplex families within the Spanish origin population of the Central Valley of Costa Rica as potential endophenotypes for genetic studies. METHODS: Individuals with a diagnosis of schizophrenia or schizoaffective disorder were compared with unaffected relatives and 15 unrelated controls with no family history of schizophrenia. RESULTS: Although the sample size is small, the results confirm previous reports in the literature of deficits in working memory, executive function, processing speed, and verbal fluency in individuals with schizophrenia compared with controls and intermediate performance in nonpsychotic family members compared with controls. We also found several suggestive quantitative cognitive trait loci with log of the odds greater than 1.75. CONCLUSION: These findings suggest that the cognitive deficits in schizophrenia are consistent aspects of the illness, although their usefulness as endophenotypes for genetic studies remains unclear. PMID- 19125109 TI - Lack of exonic sulfotransferase 4A1 mutations in controls and schizophrenia cases. AB - Sulfotransferase 4A1 (SULT4A1) is a novel sulfotransferase expressed almost exclusively in the brain. The gene is located on chromosome 22q13.3, a region implicated in predisposition to schizophrenia. Recently, a variable microsatellite region located upstream of SULT4A1 was found to be associated with an increase in schizophrenia risk. We hypothesised that if functional dysregulation of SULT4A1 was involved in the aetiology of schizophrenia, then genetic variants in the coding sequence of SULT4A1 might be identified in cases compared with controls. To test this, we carried out a mutation analysis of the coding region (exons 2-7) in 71 Australian schizophrenia cases and 69 controls. We found no mutations, either synonymous or nonsynonymous, in either cohort. However, intronic variants (IVS5+12 C>T and IVS5+28 G>C) were identified, the frequency of which was not statistically different between cases and controls. The lack of polymorphisms in the coding region of the SULT4A1 gene is highly unusual and, along with its high conservation between species, suggests that SULT4A1 may have an important function in vivo. However, our findings do not support the hypothesis that germline mutations in the coding region of SULT4A1 contribute to susceptibility to schizophrenia. PMID- 19125110 TI - No association between the D-aspartate oxidase locus and schizophrenia. PMID- 19125111 TI - Shorter dinucleotide repeat length in the DRD5 gene is associated with attention deficit hyperactivity disorder. PMID- 19125112 TI - Abstracts of the 9th Annual International Meeting on Simulation in Healthcare. January 11-14, 2009. Lake Buena Vista, Florida, USA. PMID- 19125117 TI - Weekly high-dose 5-fluorouracil as a 24-h infusion and sodium folinic acid (AIO regimen) plus irinotecan in patients with locally advanced nonresectable and metastatic adenocarcinoma or squamous cell carcinoma of the oesophagus: a phase II trial. AB - In the majority of patients with oesophageal carcinoma, curative treatment proves to be impossible when diagnosis was established; therefore, most of the patients are candidates for palliative chemotherapy. The aim of this phase II study was to evaluate the efficacy and safety of 5-fluorouracil/folinic acid (AIO regimen) plus irinotecan in patients with locally advanced or metastatic carcinoma of the oesophagus. The methods used a prospective phase II trial, start: November 2002; patients: n=25; chemotherapy: irinotecan (80 mg/m2) as a 1-h infusion and 5 fluorouracil (2000 mg/m2) with sodium folinic acid (500 mg/m2) as a 24-h infusion on days 1, 8, 15, 22, 29 and 36, repeated on day 57. Last date of evaluation: 28 February 2007; n=24; adenocarcinoma: n=13, squamous cell carcinoma (SCC): n=11; UICC III/IV: 3/21; grading G1/G2/G3/G4: 0/8/12/4; median age: 58 years (range 44 75); men/women: 19/5; Eastern Cooperative Oncology Group index 0/1/2: 3/17/4; applications: 460. Higher-grade toxicity: grade 3 diarrhoea: n=2, grade 4 diarrhoea: n=1, grade 4 vomiting: n=1, grade 4 nausea: n=1, grade 3 fatigue: n=1, grade 3 hyponatraemia: n=2, grade 4 elevation of creatinine: n=1, thrombosis of the vena subclavia: n=1, ischaemic lesion of the brain stem: n=1. Three patients died after two chemotherapeutic applications because of high tumour burden. Evaluable for response: n=19. Partial response: n=8 (33%), stable disease: n=9 (38%), progressive disease: n=2 (8%), not evaluable: n=5 (21%). Time-to progression: 6.6 months (range 1.6-24.6). Total median survival: 13.6 months (median survival of adenocarcinoma: 20.3 months, median survival of SCC: 10.0 months). Secondary resection (R0): n=3. In oesophageal carcinomas, the AIO regimen plus irinotecan is excellently manageable as an outpatient treatment and shows efficacy in adenocarcinomas and SCCs of the oesophagus. PMID- 19125118 TI - Reduced sensitivity to the locomotor-stimulant effects of cocaine is associated with increased sensitivity to its discriminative stimulus properties. AB - Outbred Long-Evans rats exhibit wide variation in their locomotor response to cocaine. Here, we investigated the relationship between these individual differences and interoceptive effects of cocaine in low cocaine responder (LCR) and high cocaine responder (HCR) phenotypes. Rats were trained to discriminate cocaine (10.0 mg/kg, intraperitoneally) from saline by repeated pairings of injections with one of two response levers. In subsequent tests for stimulus generalization to other cocaine doses (1.25-15.0 mg/kg), LCRs exhibited partial to-full generalization at 1.85 and 2.5 mg/kg cocaine, respectively, whereas HCRs did not. When the selective 5-HT reuptake inhibitor fluoxetine (5.0 mg/kg) was coadministered with saline or different cocaine doses, we observed similar upward shifts in dose-response in both phenotypes. In contrast, coadministration of the 5-HT2A/2C agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI; 0.3 mg/kg) led to partial substitution of DOI for cocaine and enhancement of the stimulus properties of 1.25 mg/kg cocaine in LCRs only. Finally, a retest of cocaine-induced locomotion after discrimination testing revealed marked behavioral sensitization in LCRs and modest changes in behavior in HCRs. Taken together, these results suggest that initial sensitivity to the locomotor stimulant effects of cocaine is inversely related to its interoceptive properties and that differences in 5-HT systems may contribute to the phenotypic differences observed. PMID- 19125119 TI - Supporting cancer patients dying at home or at a hospital for Taiwanese family caregivers. AB - Most Taiwanese patients with cancer prefer to die at home, but this preference is often unexpressed due to concern about burdening family caregivers (FCs). However, few studies have compared the caregiving burden of FCs while providing end-of-life care to a relative who eventually dies at home or hospital. This prospective, longitudinal study investigated differences in amount of assistance provided, subjective caregiving burden, depressive symptoms, and quality of life between FCs of Taiwanese patients with cancer who were terminally ill, then died at home and at a hospital. After the intake interview with 187 FCs, data were collected every 2 weeks until the patients died (M [SD] = 6.69 [4.92] times of interviews; range, 1 to 25; median, 5.0) and were analyzed by longitudinal logistic regression with a generalized estimating equation. Family caregivers whose relative died at home provided significantly more intensive assistance than their counterparts did, and caregiving more negatively impacted their finances. However, caregiving did not impact their health, daily schedule, perceived family support, or depressive symptoms to a greater negative extent. Instead, there is a trend for these caregivers to enjoy higher quality of life and to find more positive meaning in providing caregiving at home than do FCs whose relative died in hospital. In conclusion, supporting patients with cancer who are terminally ill and dying at home did not create a greater burden for Taiwanese FCs compared with those whose relatives are dying at a hospital. PMID- 19125120 TI - Vitamin D insufficiency and musculoskeletal symptoms in breast cancer survivors on aromatase inhibitor therapy. AB - Breast cancer survivors (BCSs) on aromatase inhibitor (AI) therapy often experience musculoskeletal symptoms (joint pain and stiffness, bone and muscle pain, and muscle weakness), and these musculoskeletal symptoms may be related to low serum levels of vitamin D. The primary purpose of this pilot exploratory study was to determine whether serum levels of 25-hydroxyvitamin D (25[OH]D) concentration were below normal (<30 ng/mL) in 29 BCSs on AI therapy and if musculoskeletal symptoms were related to these low vitamin D levels. The mean (SD) serum 25(OH)D level was 25.62 (4.93) ng/mL; 86% (n = 25) had levels below 30 ng/mL. Patients reported muscle pain in the neck and back, and there was a significant inverse correlation between pain intensity and serum 25(OH)D levels (r = -0.422; P < .05 [2 tailed]). This sample of BCSs taking AIs had below normal levels of serum 25(OH)D despite vitamin D supplements. This is one of the few studies to document a significant relationship between vitamin D levels and muscle pain in BCSs on AI therapy. Findings from this pilot study can be used to inform future studies examining musculoskeletal symptoms in BCSs on AI therapy and relationships with low serum levels of vitamin D. PMID- 19125122 TI - Coping with a child with acute lymphocytic leukemia: the experiences of Chinese fathers in Hong Kong. AB - Using a qualitative approach, this article aims to describe the experiences of Hong Kong Chinese fathers whose children were diagnosed with acute lymphocytic leukemia. The experiences and coping strategies used were viewed from the gender perspective. Two in-depth interviews scheduled to coincide with the disease trajectory of acute lymphocytic leukemia were conducted with 9 fathers, and data were analyzed using the matrix system described by Miles and Huberman. Four categories were identified, including fathers' initial reactions to the child's confirmed diagnosis, the decision to disclose the child's diagnosis to others, social support of the fathers, and their effective coping mechanisms. Previous research has shown that men are expected to be emotionally strong to support their spouse. Findings from this study indicate that Hong Kong Chinese fathers need emotional support especially at the onset of the child's diagnosis. Implications for healthcare professionals include the need for ongoing psychosocial support and education over the course of the child's illness. Thus, assessment of the different coping strategies used by the fathers plays a vital role in providing quality care to these fathers. Limitations of the study and recommendations for future research are also included. PMID- 19125124 TI - Challenges and opportunities for evidence-based genetics practice. PMID- 19125121 TI - Theories, models, and frameworks related to sleep-wake disturbances in the context of cancer. AB - The purpose of this article was to review theories, models, and frameworks of sleep disturbances referenced in the cancer literature. Sleep-wake disturbances in cancer are a significant problem that negatively affects quality of life. There is no previously published review of the theories, models, or frameworks used to study sleep-wake disturbances in the context of cancer. Describing existing theories or models and their application in cancer is important to advance knowledge in this area. Two theories and 9 models were identified for review. These have been used to further understand the problem of sleep-wake disturbances as a primary or secondary symptom within the cancer literature. Searches were conducted from January 1, 1970, to July 31, 2008, to find relevant articles using 4 electronic databases: MEDLINE, CINAHL, PubMed, and PsychINFO. On the basis of the search, 73 descriptive or intervention studies were identified and reviewed. Most research was atheoretical, with no identified theory, model, or framework. In studies that did use theory or models, few were applied in more than one study. Although several commonalities across models did emerge, a more comprehensive and widely used model could help guide nursing research to facilitate effective symptom management for this prominent problem in cancer. PMID- 19125125 TI - Recommendations from the EGAPP Working Group: can tumor gene expression profiling improve outcomes in patients with breast cancer? AB - SUMMARY OF RECOMMENDATIONS: The EGAPP Working Group (EWG) found insufficient evidence to make a recommendation for or against the use of tumor gene expression profiles to improve outcomes in defined populations of women with breast cancer. For one test, the EWG found preliminary evidence of potential benefit of testing results to some women who face decisions about treatment options (reduced adverse events due to low risk women avoiding chemotherapy), but could not rule out the potential for harm for others (breast cancer recurrence that might have been prevented). The evidence is insufficient to assess the balance of benefits and harms of the proposed uses of the tests. The EWG encourages further development and evaluation of these technologies. RATIONALE: The measurement of gene expression in breast tumor tissue is proposed as a way to estimate the risk of distant disease recurrence in order to provide additional information beyond current clinicopathological risk stratification and to influence decisions about treatment in order to improve health outcomes. Based on their review of the EGAPP commissioned evidence report, Impact of Gene Expression Profiling Tests on Breast Cancer Outcomes and other data summaries, the EWG found no direct evidence linking tumor gene expression profiling of women with breast cancer to improved outcomes, and inadequate evidence to construct an evidence chain. However, further evaluation on the clinical utility of some tests and management algorithms, including well-designed randomized controlled trials, is warranted. ANALYTIC VALIDITY: Some data on technical performance of assays were identified for MammaPrint and Oncotype DX, though estimates of analytic sensitivity and specificity could not be made. Published performance data on the laboratory developed Quest H:I Test were limited. Overall, the EWG found the evidence to be inadequate. CLINICAL VALIDITY: The EWG found adequate evidence regarding the association of the Oncotype DX Recurrence Score with disease recurrence and adequate evidence for response to chemotherapy. The EWG found adequate evidence to characterize the association of MammaPrint with future metastases, but inadequate evidence to assess the added value to standard risk stratification, and could not determine the population to which the test would best apply. The evidence was inadequate to characterize the clinical validity of the Quest H:I Test. CLINICAL UTILITY: The EWG found no evidence regarding the clinical utility of the MammaPrint and Quest H:I Ratio tests, and inadequate evidence regarding Oncotype DX. These technologies have potential for both benefit and harm. CONTEXTUAL ISSUES: The EWG reviewed economic studies that used modeling to predict potential effects of using gene profiling, and judged the evidence inadequate. PMID- 19125126 TI - Recommendations from the EGAPP Working Group: genetic testing strategies in newly diagnosed individuals with colorectal cancer aimed at reducing morbidity and mortality from Lynch syndrome in relatives. AB - SUMMARY OF RECOMMENDATIONS: The Evaluation of Genomic Applications in Practice and Prevention (EGAPP) Working Group found sufficient evidence to recommend offering genetic testing for Lynch syndrome to individuals with newly diagnosed colorectal cancer to reduce morbidity and mortality in relatives. We found insufficient evidence to recommend a specific genetic testing strategy among the several examined. RATIONALE: Genetic testing to detect Lynch syndrome in individuals with newly diagnosed colorectal cancer (CRC) is proposed as a strategy to reduce CRC morbidity and mortality in their relatives (see Clinical Considerations section for definition of Lynch syndrome). The EGAPP Working Group (EWG) constructed a chain of evidence that linked genetic testing for Lynch syndrome in patients with newly diagnosed CRC with improved health outcomes in their relatives. We found that assessing patients who have newly diagnosed CRC with a series of genetic tests could lead to the identification of Lynch syndrome. Relatives of patients with Lynch syndrome could then be offered genetic testing, and, where indicated, colorectal, and possibly endometrial, cancer surveillance, with the expectation of improved health outcome. The EWG concluded that there is moderate certainty that such a testing strategy would provide moderate population benefit. ANALYTIC VALIDITY: The EWG found adequate evidence to conclude that the analytic sensitivity and specificity for preliminary and diagnostic tests were high. CLINICAL VALIDITY: After accounting for the specific technologies and numbers of markers used, the EWG found at least adequate evidence to describe the clinical sensitivity and specificity for three preliminary tests, and for four selected testing strategies. These measures of clinical validity varied with each test and each strategy (see Clinical Considerations section). CLINICAL UTILITY: The EWG found adequate evidence for testing uptake rates, adherence to recommended surveillance activities, number of relatives approachable, harms associated with additional follow-up, and effectiveness of routine colonoscopy. This chain of evidence supported the use of genetic testing strategies to reduce morbidity/mortality in relatives with Lynch syndrome. Several genetic testing strategies were potentially effective, but none was clearly superior. The evidence for or against effectiveness of identifying mismatch repair (MMR) gene mutations in reducing endometrial cancer morbidity or mortality was inadequate. CONTEXTUAL ISSUES: CRC is a common disease responsible for an estimated 52,000 deaths in the United States in 2007. In about 3% of newly diagnosed CRC, the underlying cause is a mutation in a MMR gene (Lynch syndrome) that can be reliably identified with existing laboratory tests. Relatives inheriting the mutation have a high (about 45% by age 70) risk of developing CRC. Evidence suggests these relatives will often accept testing and increased surveillance. PMID- 19125127 TI - EGAPP supplementary evidence review: DNA testing strategies aimed at reducing morbidity and mortality from Lynch syndrome. PMID- 19125128 TI - Recommendations from the EGAPP Working Group: can UGT1A1 genotyping reduce morbidity and mortality in patients with metastatic colorectal cancer treated with irinotecan? AB - SUMMARY OF RECOMMENDATIONS: The Evaluation of Genomic Applications in Practice and Prevention (EGAPP) Working Group found that the evidence is currently insufficient to recommend for or against the routine use of UGT1A1 genotyping in patients with metastatic colorectal cancer who are to be treated with irinotecan, with the intent of modifying the dose as a way to avoid adverse drug reactions (severe neutropenia). RATIONALE: The EGAPP Working Group (EWG) found no intervention trials showing that targeted dosing of irinotecan based on UGT1A1 genotyping could reduce the rates of two specific adverse drug events, severe (Grade 3-4) neutropenia or diarrhea. Observational studies indicate a significant association between UGT1A1 genotypes, particularly *28/*28 and *1/*28, and the occurrence of severe neutropenia. Observational studies also indicate a possible association between severe diarrhea and these UGT1A1 genotypes, but the association is not statistically significant. An additional finding was the suggestion that reducing irinotecan dose may result in patient harms due to diminished effectiveness of treatment in highest risk individuals (*28/*28 homozygotes), and that a higher dose might be warranted among individuals at lower risk of adverse drug events (*1/*1 and *1/*28 genotypes). This review did not consider higher risk patients (e.g., having previous adverse reactions to irinotecan or additional risk factors for neutropenia). ANALYTIC VALIDITY: The EWG found adequate evidence to conclude that analytic sensitivity and specificity were high for the commonly tested alleles, but evidence was inadequate for rarer alleles. CLINICAL VALIDITY: The EWG found adequate evidence of a significant association between UGT1A1 genotype and the incidence of severe neutropenia at standard doses of irinotecan. The EWG found adequate evidence of a possible association between genotype and severe diarrhea, but the effect was smaller and not statistically significant. The EWG found adequate evidence of a significantly higher rate of tumor response to standard irinotecan dosing among individuals with the genotype at highest risk of adverse drug events (*28/*28). CLINICAL UTILITY: The EWG found no evidence to support clinical utility in the proposed clinical scenario. Preliminary modeling suggests that, even if targeted dosing were to be highly effective, it is not clear that benefits (reduced adverse drug events) outweigh harms (unresponsive tumors). CONTEXTUAL ISSUES: Addressing patient preferences regarding risk of side effects and quality of life, versus aggressive treatment to potentially improve effectiveness, is expected practice. In addition, a recent study reported that risk for neutropenia was lower at lower irinotecan doses. For treatment regimens utilizing lower irinotecan doses, UGT1A1 genotype may not be a useful indicator of risk for adverse drug events. Further rigorous evaluation of UGT1A1 genotyping using current and promising irinotecan treatment protocols is warranted. PMID- 19125129 TI - Can UGT1A1 genotyping reduce morbidity and mortality in patients with metastatic colorectal cancer treated with irinotecan? An evidence-based review. AB - This evidence-based review addresses the question of whether testing for UGT1A1 mutations in patients with metastatic colorectal cancer treated with irinotecan leads to improvement in outcomes (e.g., irinotecan toxicity, response to treatment, morbidity, and mortality), when compared with no testing. No studies were identified that addressed this question directly. The quality of evidence on the analytic validity of current UGT1A1 genetic testing methods is adequate (scale: convincing, adequate, inadequate), with available data indicating that both analytic sensitivity and specificity for the common genotypes are high. For clinical validity, the quality of evidence is adequate for studies reporting concentration of the active form of irinotecan (SN-38), presence of severe diarrhea, and presence of severe neutropenia stratified by UGT1A1 common genotypes. The strongest association for a clinical endpoint is for severe neutropenia. Patients homozygous for the *28 allele are 3.5 times more likely to develop severe neutropenia compared with individuals with the wild genotype (risk ratio 3.51; 95% confidence interval 2.03-6.07). The proposed clinical utility of UGT1A1 genotyping would be derived from a reduction in drug-related adverse reactions (benefits) while at the same time avoiding declines in tumor response rate and increases in morbidity/mortality (harms). At least three treatment options for reducing this increased risk have been suggested: modification of the irinotecan regime (e.g., reduce initial dose), use of other drugs, and/or pretreatment with colony-stimulating factors. However, we found no prospective studies that examined these options, particularly whether a reduced dose of irinotecan results in a reduced rate of adverse drug events. This is a major gap in knowledge. Although the quality of evidence on clinical utility is inadequate, two of three reviewed studies (and one published since our initial selection of studies for review) found that individuals homozygous for the *28 allele had improved survival. Three reviewed studies found statistically significant higher tumor response rates among individuals homozygous for the *28 allele. We found little or no direct evidence to assess the benefits and harms of modifying irinotecan regimens for patients with colorectal cancer based on their UGT1A1 genotype; however, results of our preliminary modeling of prevalence, acceptance, and effectiveness indicate that reducing the dose would need to be highly effective to have benefits outweigh harms. An alternative is to increase irinotecan dose among wild-type individuals to improve tumor response with minimal increases in adverse drug events. Given the large number of colorectal cancer cases diagnosed each year, a randomized controlled trial of the effects of irinotecan dose modifications in patients with colorectal cancer based on their UGT1A1 genotype is feasible and could clarify the tradeoffs between possible reductions in severe neutropenia and improved tumor response and/or survival in patients with various UGT1A1 genotypes. PMID- 19125130 TI - Cardiac tissue engineering. AB - The first 2 reviews in this series have described the defining properties of stem cells, their possible sources, and some initial attempts at their clinical use for tissue regeneration and repair. This third and final article in the series describes bioengineering methods for building physical structures to contain and organize implanted cells. The relevant theory is that appropriate physical supporting structures will help implanted cardiac stem cell populations organize themselves into functioning cardiac tissue that integrates physically and functionally with the receiving heart. The purpose of cardiac tissue engineering is to replace or repair injured heart muscle effectively. Supporting materials to create habitable spaces can provide the basic requirements of cardiac muscle cells. The design of such supporting materials influences the behavior of cells; the shape, dimensions, and chemistry of substrates affect such processes as attachment, cell signaling, and differentiation. As cardiac muscle cells flourish in artificial environments, they may become functional tissue with clinical value. This review summarizes the major bioengineering approaches for containing and organizing cardiac muscle cells and their potential to ameliorate total heart failure. PMID- 19125131 TI - Alternative coping strategies and decision delay in seeking care for acute myocardial infarction. AB - BACKGROUND AND OBJECTIVE: In attempting to manage acute myocardial infarction (AMI) symptoms, individuals often engage in coping strategies that cause them to delay seeking timely medical care. Review of the literature revealed that several coping strategies were examined in relation to AMI delay, but there were no studies investigating the extent of use of each coping strategy. The purpose of this study was to examine the extent of use of 15 coping strategies and their associations with the time taken to decide to seek medical care. SUBJECTS AND METHODS: Secondary data analyses were performed on a sample of 135 patients with AMI from Canada and the United States from whom data were collected via structured interviews. Descriptive analyses were used to identify the extent of use of alternative coping strategies. Spearman rho was used to test the associations of these strategies with decision delay. RESULTS AND CONCLUSIONS: The 3 most frequently used coping strategies were trying to relax, wishing/praying for symptoms to disappear, and discussing symptoms with someone. Nine coping strategies were significantly associated with decision delay. The findings of this study are useful in teaching patients about avoiding the use of coping strategies when faced with AMI symptoms. This can potentially help reduce delays in seeking care for AMI, which will result in better health outcomes for patients with AMI. PMID- 19125132 TI - Tumor-induced osteomalacia: successful treatment by radio-guided tumor surgery. AB - Tumor-induced osteomalacia is a rare syndrome characterized by urinary phosphate loss with hypophosphatemic osteomalacia. The proposed pathogenetic mechanism is paraneoplastic secretion of phosphaturic factors (so-called phosphatonins).We describe a 34-year-old male patient who presented with severe pain of the spine and ribs for at least 2 years. Bone scintigraphy using Technetium hydroxymethylene diphosphonate (Tc HDP) showed multiple lesions suggesting metastatic disease. Bone biopsy however revealed osteomalacia. The patient had subnormal plasma phosphate levels (0.42 mmol/L; normal range, 0.87-1.45) and markedly increased phosphate clearance (82.8 mL/min; normal range, 5.4-16.2). The patient was treated with phosphate supplementation (up to 5 g daily) along with calcium (1000 mg daily) and calcitriol (1.5 microg daily). Although this therapy did not correct hypophosphatemia, it resulted in complete relief of pain within several months. (111)In pentetreotide scintigraphy showed a tiny lesion of 1-cm diameter, which could be localized to the left femoral neck in close vicinity to the greater trochanter by MRI and image fusion analysis. This lesion had not been visualized by Tc-99m HDP bone scintigraphy. Intraoperatively, use of a hand-held gamma probe after administration of (111)Indium pentetreotide ((111)In pentetreotide) clearly identified the tumor, which was completely removed and was shown to be a hemangiopericytoma. After removal of the tumor, phosphate metabolism normalized within 1 week without requirement of phosphate supplementation. Hypophosphatemic osteomalacia, although rare, raises an important differential diagnosis. An underlying tumor may be detected only by (111)In pentetreotide scintigraphy. Preoperative labeling with (111)In pentetreotide is a useful tool in detecting these tumors during surgery.This 34 year old man with osteomalacia had a small causative hemangiopericytoma detected in the indium pentetreotide scintography. PMID- 19125133 TI - Development of antibodies against humanized antitumor necrosis factor and loss of efficacy in a black spider monkey with inflammatory bowel disease. PMID- 19125134 TI - Neurogenic bladder in a patient with systemic lupus erythematosus and cerebral involvement. PMID- 19125135 TI - The effect of serum urate on gout flares and their associated costs: an administrative claims analysis. AB - BACKGROUND: Increased serum urate (sUA) levels (> or =6.0 mg/dL) are associated with increased likelihood of acute gout attacks, or "flares." OBJECTIVES: Identify gout flares with administrative claims data; examine the relationship between sUA and flares; examine the association between sUA and flare-related costs. METHODS: This retrospective administrative claims analysis examined subjects with gout (> or =2 medical claims with ICD-9-CM diagnosis code 274.xx or > or =1 claim with a gout diagnosis and > or =1 pharmacy claim for allopurinol, probenecid, colchicine, or sulfinpyrazone) between January 1, 2002 and March 31, 2004. Each subject was observed during 1-year baseline and 1-year follow-up periods. Gout flares were identified with an algorithm using claims for services associated with flares. Outcomes were sUA (mg/dL) and flare-related health care costs. Logistic regression examined the likelihood of flare; generalized linear modeling regression measured the impact of baseline sUA on flare costs, controlling for demographic and health status variables. RESULTS: The study sample comprised 18,243 subjects with mean age of 53.9 years. sUA was available for 4277 (23%) subjects. Sixty-two percent (11,253) of subjects had > or =1 flare. The number of mean, unadjusted flares increased with sUA. Logistic results showed subjects with baseline sUA > or =6.0 relative to sUA <6.0 had 1.3 times the odds of gout flare (P <0.05). Generalized linear modeling results showed that baseline sUA > or =6.0 was associated with 2.1 to 2.2 times higher flare costs than was baseline sUA <6.0 (P <0.05). CONCLUSIONS: sUA was a significant predictor both of gout flare and related costs. This highlights the importance of gout management strategies aimed at controlling sUA. PMID- 19125136 TI - Clinical diagnostic criteria for gout: comparison with the gold standard of synovial fluid crystal analysis. AB - BACKGROUND: Several classification or diagnostic criteria sets for gout have been proposed but none validated. OBJECTIVE: This small pilot study considered urate crystal identification as the gold standard for diagnosis and compared the clinical aspects of 3 proposed criteria sets with that standard. METHODS: Eighty two subjects who had synovial fluid analyses in a VA medical center were studied. ARA (ACR), Rome, and New York clinical criteria sets and individual criteria were recorded in the 30 patients who had urate crystals versus the remainder with no urate crystals. RESULTS: Presence of 2 of 3 Rome clinical criteria had the highest positive predictive value at 76.9%. None of the 3 studied criteria sets were more than 70% sensitive or 88.5% specific. The clinical features of the ARA (ACR) preliminary classification criteria had 70% sensitivity and 78.8% specificity. CONCLUSIONS: The various proposed clinical criteria can provide support for a diagnosis or exclusion of gout, but unless improved criteria can be developed crystal identification should remain the gold standard. PMID- 19125137 TI - Fibromyalgia is associated with impaired balance and falls. AB - BACKGROUND/OBJECTIVE: The purpose of this study was to determine whether fibromyalgia (FM) patients differ from matched healthy controls in clinical tests of balance ability and fall frequency. METHODS: Thirty-four FM patients and 32 age-matched controls were administered the Balance Evaluation-Systems Test (BESTest), rated their balance confidence with the Activities-Specific Balance Confidence (ABC) Scale, and reported the number of falls in the last 6 months. The Fibromyalgia Impact Questionnaire was used to assess FM severity. RESULTS: FM patients had significantly impaired balance in all components of the BESTest compared with controls. They also scored more poorly on balance confidence. Overall FM severity (Fibromyalgia Impact Questionnaire) correlated significantly with the BESTest and the ABC scale. The BESTest and ABC correlated significantly with 6 commonly reported FM symptoms (excluding pain). FM patients reported a total of 37 falls over the last 6-months compared with 6 falls in healthy controls. CONCLUSION: FM is associated with balance problems and increased fall frequency. Patients were aware of their balance problems. These results suggest that FM may affect peripheral and/or central mechanisms of postural control. Further objective study is needed to identify the relative contributions of various neural and musculoskeletal and other impairments to postural stability in FM to provide clinicians with methods to maximize postural stability and help fall prevention. PMID- 19125138 TI - Silicone synovitis: longer term outcome data and review of the literature. AB - BACKGROUND: Silicone had been popular for small and medium joint prostheses from the 1960s until the 1980s when concerns about subsequent arthritis arose. There are still patients with long-standing silicone prostheses. We describe 7 cases of synovitis attributed to these. METHODS: This is a case series of patients having synovitis in the vicinity of a silicone arthroplasty. The diagnosis was determined by examination and, radiographic and operative characteristics of the joint. Underlying diseases of the patients, treatment strategies, and responses to treatment were reviewed. RESULTS: Seven charts were reviewed. The ages ranged from 53 to 90 years (mean = 71.5). Two patients had osteoarthritis, 1 had rheumatoid arthritis, 3 had trauma, and 1 had avascular necrosis. Joints included 2 metatarsal phalangeal, 3 carpal bones, 1 radial head, and 2 first carpal metacarpals. The time from arthroplasty to synovitis was 9 to 30 years (mean = 16). Five patients had surgical management and 2 had medical management. Four of the surgically managed patients had resolution of their symptoms, 1 had a recurrent synovitis requiring further surgery. One person had 2 arthroplasties, but developed synovitis at only 1 site. The 2 patients medically managed continued to have synovitis. CONCLUSIONS: Synovitis continues to be a consequence of silicone arthroplasty. It can be a late complication with our shortest time to synovitis being 9 years. Underlying pathology does not seem to predict the synovitis as patients with osteoarthritis, rheumatoid arthritis, avascular necrosis, and trauma are all represented. Surgically managed patients did well, whereas patients managed medically did not have resolution of their symptoms. PMID- 19125139 TI - Restless legs syndrome in a rheumatoid arthritis patient cohort. AB - OBJECTIVE: To use the 2003 International Restless Legs Syndrome Study Group (IRLSSG) diagnostic criteria and to evaluate restless legs syndrome (RLS) prevalence in a rheumatoid arthritis (RA) and osteoarthritis (OA) population. Further, we wished to evaluate physician awareness of this disorder by as reflected in prevalence of preexisting diagnoses of RLS in these populations. METHODS: This was a questionnaire study of Saskatchewan RA and OA patients enrolled in a longitudinal database study. A data collection instrument, including the 2003 IRLSSG criteria for RLS was distributed to the patients enrolled. RESULTS: Of the 193 respondents, 158 (81.9%) were women. The population consisted of 148 RA and 45 OA patients. RA patients were younger (mean age, 65.8 years) in comparison with those in the OA group (mean age, 72.8 years; P < 0.001). All criteria for RLS were met by 27.7% of RA patients and by 24.4% of OA patients. A previous diagnosis of RLS was reported by 2.6% of patients. CONCLUSIONS: A quarter of all our patients met the 2003 IRLSSG criteria, in both RA and OA groups; however, only 2.6% of study patients reported a previous diagnosis of RLS. As RLS can significantly affect quality of life, increased awareness with improvement in surveillance, recognition, and treatment would be beneficial to patient care. We advocate screening for symptoms of sleep disorders to be incorporated into the routine rheumatologic history for all patients with RA and OA. PMID- 19125140 TI - Responding to the burden of STD, HIV, and viral hepatitis in correctional populations through program collaboration and integration. PMID- 19125141 TI - A 15-minute interactive, computerized condom use intervention with biological endpoints. AB - BACKGROUND: Brief face-to-face-behavioral interventions have been shown to be efficacious, but are costly to sustain and to widely disseminate. This study evaluated the efficacy of a 15-minute theory-based behavioral intervention designed to increase condom use and reduce new cases of Neisseria gonorrhoeae and Chlamydia trachomatis. METHODS: Participants were randomly assigned via the computer to the intervention or the comparison group stratified by gender and their baseline stage of change (motivational readiness) for using condoms consistently (100%) with their main partners. Behavioral data and biologic specimens for testing of Neisseria gonorrhoeae and Chlamydia trachomatis were obtained at baseline and at 6 months post intervention. The intervention was delivered via an audio, multimedia, computerized application that provided individualized interventions to patients based on their responses to assessment items; comparison patients interacted with a 15-minute, computerized, multiple health risk assessment with no intervention. RESULTS: The majority of the sample (N = 430) was black (88%); 54.5% women; with a mean age = 24.5. Assuming all participants who did not return to the clinic at 6 months were not using condoms consistently, 32% of the treatment group versus 23% in the comparison group reported consistent condom use (P = 0.03). The combined Neisseria gonorrhoeae and Chlamydia trachomatis incidence declined to 6% in the intervention group versus 13% in the comparison group (P = 0.04). Results from a regression analysis revealed that the only statically significant predictor of sexually transmitted diseases infection at the follow-up was group assignment (OR = 1.91, 95% confidence index = 1.09-3.34; P = 0.043). CONCLUSIONS: These findings suggest that brief, interactive, computer-delivered interventions provided at the evaluation visit increase condom use and reduce sexually transmitted diseases without putting additional burden on clinicians or staff. PMID- 19125142 TI - Rapidly increasing prevalence of HIV and syphilis and HIV-1 subtype characterization among men who have sex with men in Jiangsu, China. AB - OBJECTIVES: To investigate the prevalence of HIV, hepatitis B (HBV), hepatitis C (HCV), and syphilis among men who have sex with men (MSM) in 2 cities of Jiangsu, China, and to characterize the HIV-1 subtypes prevalent among this population. METHODS: During September 2006 and July 2007, 296 and 173 MSM were recruited from Nanjing and Yangzhou, respectively. Sera samples were collected and tested for HIV, HBV, HCV, and syphilis infections. The nucleotide sequences of p17 and C2V3 regions were determined by RT-nested-PCR and sequencing. HIV-1 subtypes were characterized by phylogenetic analysis. RESULTS: The prevalence of HIV, HBV, HCV, and syphilis infections among MSM was 5.8%, 11.1%, 0.7%, and 27.7%, respectively. The prevalence of HIV and syphilis was significantly higher in 2006-2007 than in 2003 (P 0.05). The phylogenetic tree of p17 showed that HIV 1 subtypes B, CRF01_AE, and CRF07_BC accounted for 35.7%, 35.7%, and 28.6%, respectively. The result of C2V3 showed that 45.5%, 36.4%, and 18.2% sequences belonged to HIV-1 subtype B, CRF01_AE, and BC recombinants, respectively. The subtype characterization in Jiangsu was significantly different from those in Beijing (P <0.05). Furthermore, Jiangsu HIV-1 B strains were different from majority of China B' strains and originated from Beijing. CONCLUSIONS: The rapidly increasing prevalence and complex subtypes of HIV-1 suggest that effective prevention and intervention strategies are urgently needed for MSM in Jiangsu. PMID- 19125143 TI - Ethnic/racial homogeneity and sexually transmitted disease: a study of 77 Chicago community areas. AB - BACKGROUND: Sexually transmitted diseases (STDs) remain an intractable public health problem in the United States. Wide ethnic and racial disparities persist in the rates of reported STDs. Blacks and Hispanics have higher rates of chlamydia and gonorrhea than European Americans. METHODS: With data collected in 2002 from 77 well defined communities in Chicago, we examined the association of ethnic/racial homogeneity of the communities and incidence of chlamydia and gonorrhea. The multivariate regression models controlled for other sociodemographic variables. RESULTS: Communities where at least 60% of the residents were black had significantly higher incidence rates of these 2 STDs compared with communities where at least 60% of the residents were Hispanic. Independent of ethnic/racial homogeneity of the community, the incidence of these STDs was higher in communities that had a larger percentage of persons living in poverty, a larger percentage unemployed, fewer high school graduates, and more residents between the ages of 15 and 44. CONCLUSION: The challenge for public health authorities is to consider policy options that respond not only to sexual risk behaviors, but also to the contextual attitudes, cultural traditions, and norms, and social circumstances in ethnically homogeneous communities that may affect the spread of STDs in disadvantaged urban populations. PMID- 19125144 TI - Lymphogranuloma venereum in Portugal: unusual events and new variants during 2007. AB - BACKGROUND: Several European countries identified an ongoing LGV outbreak, particularly among men who have sex with men (MSM). In Portugal, no particular surveillance measures were launched. Nonetheless, circulating LGV strains could eventually be detected through the routine Chlamydia trachomatis ompA genotyping procedure held in the Portuguese National Institute of Health (NIH). METHODS: During 2007, 178 Chlamydia trachomatis specimens were genotyped through amplification and automated-sequencing of ompA. Sequences of 891bp (nt142-nt1032) were aligned with currently available chlamydial sequences from GenBank to identify the corresponding genotype. RESULTS: Eight Chlamydia trachomatis specimens matched LGV-genotypes (7 "L2" and 1 mixed E+L2 undetermined variant). These specimens were identified in samples collected from 4 women and 4 men. One HIV(+) MSM presented LGV related symptoms, while the other infected persons were either asymptomatic or presented no clear LGV symptoms. All samples revealed ompA sequences different from the L2/434 reference strain and from the L2b/144276, which is the most frequently described genotype during the recent LGV outbreak. CONCLUSIONS: The detection of 7 LGV specimens during 2007 in contrast with their absence over the previous 5 years. The LGV infected individuals do not seem to be related to any sexual networks of MSM, contrarily to those described in other European countries. Moreover, all Lisbon LGV specimens revealed unusual ompA sequences that differentiate them from the currently reported LGV infections in Europe. The results of the current study further justify an attentive surveillance of LGV strains infecting different populations and the study of their relation with clinical aspects and disease patterns. PMID- 19125145 TI - An epidemiological survey of chlamydial and gonococcal infections in a Canadian arctic community. AB - BACKGROUND: Sexually transmitted infections are leading causes of morbidity for Canadian Aboriginal women. To date, very few initiatives have been successful in screening, treating, and limiting these infections among these populations. OBJECTIVES: To evaluate the efficacy of universal screening, treatment and contact tracing as a means of capturing a more accurate count of chlamydia and gonorrhea prevalence and limiting transmission among Inuit communities. METHODS: 181 participants were screened for chlamydia and gonorrhea and interviewed in a cross-sectional survey (Aug-Sept/03). Information was collected on demographics, use of health services, sexual histories and STI knowledge among others. A random sample (n = 100) from the cross-sectional group was selected for the longitudinal cohort. Individuals were followed every two months post baseline for four visits (Oct/03-May/04). At each visit, participants were screened for chlamydia/gonorrhea. All positive cases and their partners were treated and contact tracing completed. Logistic Regression analysis and the McNemar Test of Correlated Proportions were used to analyze the data. RESULTS: Overall, 35 cases of chlamydia were detected, with 21 detected at baseline and 14 during follow-up. The baseline prevalence was 11.6% in comparison with 2.7% that was previously estimated. No gonorrhea was detected. The strongest factor associated with a positive chlamydia was having recent STI (OR 9.82, CI: 2.70, 35.77). CONCLUSIONS: Consistent with the literature, the results support the use of universal screening followed by prompt treatment and contact tracing in populations with greater than 10% chlamydia prevalence. PMID- 19125146 TI - Sexually transmitted infections and HIV prevalence among incarcerated men who have sex with men, 2000-2005. AB - OBJECTIVE: : Screening incarcerated populations, particularly men who have sex with men (MSM), for the identification, treatment, and prevention of sexually transmitted infections (STI) and HIV provides an effective way to access a hard to-reach, high-risk population. GOAL: : To describe findings from a screening program designed to identify STIs and HIV among incarcerated MSM. STUDY DESIGN: : The Los Angeles County Sexually Transmitted Disease Program implemented a voluntary HIV and STI screening program in the segregated MSM unit of the Los Angeles County Men's Jail in March 2000. This analysis reports on data collected through December 2005. RESULTS: : Between March 2000 and December 2005, a total of 7004 inmates participated in the screening program. The overall positivity rate for chlamydia was 3.1% (127 of 4157) and 1.7% (69 of 4106) for gonorrhea. In addition, early syphilis was identified in 1.6% of inmates (95 of 6008) and the overall prevalence of HIV was 13.4% (625 of 4658). The level of repeat testing was relatively high with 15% (1048) of inmates repeatedly incarcerated and screened for STIs over the 5-year period. Although the seroprevalence of HIV was not significantly different between repeaters and nonrepeaters, 33 inmates were HIV seropositive after having tested negative at prior bookings, resulting in an HIV incidence of 1.9%. CONCLUSIONS: : Screening incarcerated MSM in Los Angeles revealed a high prevalence of STI and HIV infection. These inmates not only represent a high-risk group, but also a unique opportunity for the identification, treatment, and counseling of this hard-to-reach, high-risk population. PMID- 19125147 TI - Sex and age correlates of Chlamydia prevalence in adolescents and adults entering correctional facilities, 2005: implications for screening policy. AB - OBJECTIVES: To evaluate sex and age correlates of chlamydia prevalence in incarcerated populations. METHODS: Cross-sectional analysis of chlamydia prevalence by demographic characteristics from incarcerated females and males entering selected juvenile and adult correctional facilities (jails) in the United States in 2005. RESULTS: A total of 97,681 and 52,485 incarcerated persons aged >/=12 years were screened for chlamydia in 141 juvenile and 22 adult correctional facilities, respectively. Overall, chlamydia prevalence was high in females (14.3% and 7.5%) in both juvenile and adult facilities when compared with that in males (6.0% and 4.6%). The chlamydia prevalence was higher in incarcerated females than in incarcerated males for persons 40 years; in males it was 8.8% in 18- to 20-year olds compared with 1.4% in those >40 years. CONCLUSIONS: The consistently high chlamydia prevalence among females in juvenile facilities and females (or=18 years) in the 2006 Behavioral Risk Factor Surveillance System. Current depression was assessed by the Patient Health Questionnaire-8 diagnostic algorithm. Lifetime diagnoses of depression, anxiety and ORCs were self-reported. The prevalence of the three psychiatric disorders was age standardized to the 2000 US population. Multivariate-adjusted prevalence ratios were computed to test associations of depression and anxiety with BMI using SUDAAN software. RESULTS: The age-adjusted prevalence of current depression, lifetime diagnosed depression and anxiety varied significantly by gender. Within each gender, the prevalence of the three psychiatric disorders was significantly higher in both men and women who were underweight (BMI<18.5 kg/m(2)), in women who were overweight (BMI: 25-<30 kg/m(2)) or obese (BMI>or=30 kg/m(2)), and in men who had class III obesity (BMI>or=40 kg/m(2)) than in those with a normal BMI (18.5-<25 kg/m(2)). After adjusting for demographics, ORCs, lifestyle or psychosocial factors, compared with men with a normal BMI, men with a BMI>or=40 kg/m(2) were significantly more likely to have current depression or lifetime diagnosed depression and anxiety; men with a BMI<18.5 kg/m(2) were also significantly more likely to have lifetime diagnosed depression. Women who were either overweight or obese were significantly more likely than women with a normal BMI to have all the three psychiatric disorders. CONCLUSIONS: Our results demonstrate that disparities in the prevalence of depression and anxiety exist among people with different BMI levels independent of their disease status or other psychosocial or lifestyle factors. PMID- 19125164 TI - Evolving the discussion. PMID- 19125165 TI - The Hsp90 mosaic: a picture emerges. PMID- 19125166 TI - Eye on RNA unwinding. PMID- 19125167 TI - RNA stability: is it the endo' the world as we know it? PMID- 19125168 TI - Chromodomain-mediated spreading on active genes. PMID- 19125169 TI - Alternative splicing: regulation without regulators. PMID- 19125171 TI - Functional characterization of the human-specific (type II) form of kallikrein 8, a gene involved in learning and memory. AB - Kallikrein 8 (KLK8) is a serine protease functioning in the central nervous system, and essential in many aspects of neuronal activities. Sequence comparison and gene expression analysis among diverse primate species identified a human specific splice form of KLK8 (type II) with preferential expression in the human brain, which may contribute to the origin of human cognition. To gain insights into the physiological and biochemical role of this novel form, we conducted functional analyses of human type II KLK8. Our results show that type II KLK8 is abundantly expressed in human embryonic stem cells and in embryo brain samples, suggesting a potential role in embryogenesis. There are dramatic expression variations in different individuals and brain regions, which is a reflection of its dynamic role in neural activities. Furthermore, the transcription start site (TSS) of KLK8 is tissue-specific, with a brain-specific TSS found in humans indicating functional specialization. Our in vitro biochemical assay shows that there is a type II-specific intermediate protein form, although the processed end point enzymes are the same for both type I and type II KLK8, suggesting that the emergence of type II KLK8 in the human brain likely leads to functional modifications of KLK8. PMID- 19125172 TI - Mefloquine--an aminoalcohol with promising antischistosomal properties in mice. AB - BACKGROUND: The treatment and control of schistosomiasis, an often neglected tropical disease that exacerbates poverty, depends on a single drug, praziquantel. The large-scale use of praziquantel might select for drug-resistant parasites, hence there is a need to develop new antischistosomal compounds. Here, we report that the antimalarial drug mefloquine possesses promising antischistosomal properties in mice. METHODOLOGY/PRINCIPAL FINDINGS: A single dose of mefloquine (200 or 400 mg/kg) administered orally to mice infected with adult Schistosoma mansoni or adult S. japonicum resulted in high or complete total and female worm burden reductions (72.3%-100%). Importantly, high worm burden reductions were also observed for young developing stages of S. mansoni and S. japonicum harbored in the mouse. Both mefloquine erythro-enantiomers resulted in high and comparable total and female worm burden reductions when given to mice with either a sub-patent or patent S. mansoni infection. CONCLUSIONS/SIGNIFICANCE: Our findings hold promise for the development of a novel antischistosomal drug based on an aminoalcohol functionality. Further in vitro and in vivo studies have been launched to elucidate the possible mechanism of action and to study the effect of mefloquine on S. haematobium and other trematodes. It will be interesting to investigate whether mefloquine, which is widely and effectively used for the treatment of malaria, has an impact on schistosomiasis in areas where both malaria and schistosomiasis co-exist. PMID- 19125173 TI - The effect of insecticide synergists on the response of scabies mites to pyrethroid acaricides. AB - BACKGROUND: Permethrin is the active component of topical creams widely used to treat human scabies. Recent evidence has demonstrated that scabies mites are becoming increasingly tolerant to topical permethrin and oral ivermectin. An effective approach to manage pesticide resistance is the addition of synergists to counteract metabolic resistance. Synergists are also useful for laboratory investigation of resistance mechanisms through their ability to inhibit specific metabolic pathways. METHODOLOGY/PRINCIPAL FINDINGS: To determine the role of metabolic degradation as a mechanism for acaricide resistance in scabies mites, PBO (piperonyl butoxide), DEF (S,S,S-tributyl phosphorotrithioate) and DEM (diethyl maleate) were first tested for synergistic activity with permethrin in a bioassay of mite killing. Then, to investigate the relative role of specific metabolic pathways inhibited by these synergists, enzyme assays were developed to measure esterase, glutathione S-transferase (GST) and cytochrome P450 monooxygenase (cytochrome P450) activity in mite extracts. A statistically significant difference in median survival time of permethrin-resistant Sarcoptes scabiei variety canis was noted when any of the three synergists were used in combination with permethrin compared to median survival time of mites exposed to permethrin alone (p<0.0001). Incubation of mite homogenates with DEF showed inhibition of esterase activity (37%); inhibition of GST activity (73%) with DEM and inhibition of cytochrome P450 monooxygenase activity (81%) with PBO. A 7-fold increase in esterase activity, a 4-fold increase in GST activity and a 2-fold increase in cytochrome P450 monooxygenase activity were observed in resistant mites compared to sensitive mites. CONCLUSIONS: These findings indicate the potential utility of synergists in reversing resistance to pyrethroid-based acaricides and suggest a significant role of metabolic mechanisms in mediating pyrethroid resistance in scabies mites. PMID- 19125174 TI - Acceptance Speech for the John McMillan Mennell Service Award of the American Academy of Orthopaedic Manual Physical Therapy. PMID- 19125175 TI - To run or not to run: a post-meniscectomy qualitative risk analysis model for osteoarthritis when considering a return to recreational running. AB - The increased likelihood of osteoarthritic change in the tibiofemoral joint following meniscectomy is well documented. This awareness often leads medical practitioners to advise patients previously engaged in recreational running who have undergone meniscectomy to cease all recreational running. This literature review examines the following questions: 1) Is there evidence to demonstrate that runners, post-meniscectomy, incur a great enough risk for early degenerative OA to cease all running? 2) Does the literature yield risk factors for early OA that would guide a physical therapist with regard to advising the post-meniscectomy patient contemplating a return to recreational running? Current literature related to meniscal structure and function, etiology and definition of osteoarthritis, methods for assessing osteoarthritis, relationship between running and osteoarthritis, and relationship between meniscectomy and osteoarthritis are reviewed. This review finds that while the probability for early osteoarthritis in the post-meniscectomy population is substantial, it is a probability and not a certainty. To help guide a physical therapist with regard to advising the patient for a safe return to running following a meniscectomy, a qualitative risk assessment based on identified risk factors for osteoarthritis in both the running and the post-meniscectomy populations is proposed. PMID- 19125176 TI - Chlamydia trachomatis serology in women with and without ovarian cancer. AB - Pelvic inflammation has been implicated in the genesis of ovarian cancer. We conducted serologic measurements of Chlamydia trachomatis antibodies as a surrogate marker of chlamydial pelvic inflammatory disease. Women with ovarian cancer (n = 521) and population-based controls (n = 766) were tested. IgG antibodies to serovar D of chlamydia elementary bodies (EBs) were detected using an ELISA assay. The odds of having ovarian cancer among women with the highest titers (>or=0.40 OD units) were 0.6 (95% CI 0.4-0.9). These data do not support our earlier finding of elevated titers for antibodies to C. trachomatis among women with ovarian cancer. PMID- 19125178 TI - SVD-based evaluation of multiplexing in multipinhole SPECT systems. AB - Multipinhole SPECT system design is largely a trial-and-error process. General principles can give system designers a general idea of how a system with certain characteristics will perform. However, the specific performance of any particular system is unknown before the system is tested. The development of an objective evaluation method that is not based on experimentation would facilitate the optimization of multipinhole systems. We derive a figure of merit for prediction of SPECT system performance based on the entire singular value spectrum of the system. This figure of merit contains significantly more information than the condition number of the system, and is therefore more revealing of system performance. This figure is then compared with simulated results of several SPECT systems and is shown to correlate well to the results of the simulations. The proposed figure of merit is useful for predicting system performance, but additional steps could be taken to improve its accuracy and applicability. The limits of the proposed method are discussed, and possible improvements to it are proposed. PMID- 19125179 TI - BPH procedural treatment: the case for value-based pay for performance. AB - The concept of "pay for performance" (P4P) applied to the practice of medicine has become a major foundation in current public and private payer reimbursement strategies for both institutional and individual physician providers. "Pay for performance" programs represent a substantial shift from traditional service based reimbursement to a system of performance-based provider payment using financial incentives to drive improvements in the quality of care. P4P strategies currently embody rudimentary structure and process (as opposed to outcomes) metrics which set relatively low-performance thresholds. P4P strategies that align reimbursement allocation with "free market" type shifts in cognitive and procedural care using evidence-based data and positive reinforcement are more likely to produce large-scale improvements in quality and cost efficiency with respect to clinical urologic care. This paper reviews current paradigms and, using BPH procedural therapy outcomes, cost, and reimbursement data, makes the case for a fundamental change in perspective to value-based pay for performance as a reimbursement system with the potential to align the interests of patients, physicians, and payers and to improve global clinical outcomes while preserving free choice of clinically efficacious treatments. PMID- 19125177 TI - Clinical Use of PPARgamma Ligands in Cancer. AB - The role of PPARgamma in adipocyte differentiation has fueled intense interest in the function of this steroid nuclear receptor for regulation of malignant cell growth and differentiation. Given the antiproliferative and differentiating effects of PPARgamma ligands on liposarcoma cells, investigation of PPARgamma expression and ligand activation in other solid tumors such as breast, colon, and prostate cancers ensued. The anticancer effects of PPARgamma ligands in cell culture and rodent models of a multitude of tumor types suggest broad applicability of these agents to cancer therapy. This review focuses on the clinical use of PPARgamma ligands, specifically the thiazolidinediones, for the treatment and prevention of cancer. PMID- 19125180 TI - Peripheral mononuclear cell resistin mRNA expression is increased in type 2 diabetic women. AB - Resistin has been shown to cause insulin resistance and to impair glucose tolerance in rodents, but in humans its physiological role still remains elusive. The aim of this study was to examine whether resistin mRNA expression in human peripheral mononuclear cells (PBMCs) and its corresponding plasma levels are altered in type 2 diabetes. Resistin mRNA levels were easily detectable in human PBMC, and found to be higher in DM2 compared to healthy women (P = .05). Similarly, mononuclear mRNA levels of the proinflammatory cytokines IL-1beta, TNF alpha, and IL-6 were all significantly higher in DM2 compared to control women (P < .001). The corresponding plasma resistin levels were slightly, but not significantly, increased in DM2 women (P = .051), and overall, they correlated significantly with BMI (r = 0.406, P = .010) and waist circumference (r = 0.516, P = .003), but not with fasting insulin levels or HOMA-IR. Resistin mRNA expression is increased in PBMC from DM2 women, together with increased expression of the inflammatory cytokines IL-1beta, TNF-alpha, and IL-6, independent of obesity. These results suggest that resistin and cytokines might contribute to the low-grade inflammation and the increased atherogenic risk observed in these patients. PMID- 19125182 TI - Improved recovery of exfoliated colonocytes from feces using newly developed immunomagnetic beads. AB - We demonstrated the feasibility of a new methodology for isolating colonocytes from feces. To reduce costs and improve the recovery rate of colonocytes from feces, we attempted to develop new immunomagnetic beads. Several sizes of magnetic beads were prepared and tagged with a monoclonal antibody against EpCAM. We made several new monoclonal antibodies against EpCAM, and each monoclonal antibody was tagged to the magnetic beads. In the simulation, the most efficient recovery of HT-29 cells was obtained using the smallest size of beads. Also, beads tagged with a monoclonal antibody with a higher affinity against EpCAM had a higher recovery rate. Similar results were obtained when the smallest size of beads with the highest-affinity monoclonal antibody was applied to clinical samples. The newly developed immunomagnetic beads may be useful for isolating colorectal cancer cells from feces, enabling the cytological or molecular biological diagnosis of CRC. PMID- 19125183 TI - Integrating biosystem models using waveform relaxation. AB - Modelling in systems biology often involves the integration of component models into larger composite models. How to do this systematically and efficiently is a significant challenge: coupling of components can be unidirectional or bidirectional, and of variable strengths. We adapt the waveform relaxation (WR) method for parallel computation of ODEs as a general methodology for computing systems of linked submodels. Four test cases are presented: (i) a cascade of unidirectionally and bidirectionally coupled harmonic oscillators, (ii) deterministic and stochastic simulations of calcium oscillations, (iii) single cell calcium oscillations showing complex behaviour such as periodic and chaotic bursting, and (iv) a multicellular calcium model for a cell plate of hepatocytes. We conclude that WR provides a flexible means to deal with multitime-scale computation and model heterogeneity. Global solutions over time can be captured independently of the solution techniques for the individual components, which may be distributed in different computing environments. PMID- 19125181 TI - PPARs Mediate Lipid Signaling in Inflammation and Cancer. AB - Lipid mediators can trigger physiological responses by activating nuclear hormone receptors, such as the peroxisome proliferator-activated receptors (PPARs). PPARs, in turn, control the expression of networks of genes encoding proteins involved in all aspects of lipid metabolism. In addition, PPARs are tumor growth modifiers, via the regulation of cancer cell apoptosis, proliferation, and differentiation, and through their action on the tumor cell environment, namely, angiogenesis, inflammation, and immune cell functions. Epidemiological studies have established that tumor progression may be exacerbated by chronic inflammation. Here, we describe the production of the lipids that act as activators of PPARs, and we review the roles of these receptors in inflammation and cancer. Finally, we consider emerging strategies for therapeutic intervention. PMID- 19125184 TI - Dilated cardiomyopathy with increased SR Ca2+ loading preceded by a hypercontractile state and diastolic failure in the alpha(1C)TG mouse. AB - Mice over-expressing the alpha(1)_subunit (pore) of the L-type Ca2+ channel (alpha(1C)TG) by 4 months (mo) of age exhibit an enlarged heart, hypertrophied myocytes, increased Ca2+ current and Ca2+ transient amplitude, but a normal SR Ca2+ load. With advancing age (8-11 mo), some mice demonstrate advanced hypertrophy but are not in congestive heart failure (NFTG),while others evolve to frank dilated congestive heart failure (FTG). We demonstrate that older NFTG myocytes exhibit a hypercontractile state over a wide range of stimulation frequencies, but maintain a normal SR Ca2+ load compared to age matched non transgenic (NTG) myocytes. However, at high stimulation rates (2-4 Hz) signs of diastolic contractile failure appear in NFTG cells. The evolution of frank congestive failure in FTG is accompanied by a further increase in heart mass and myocyte size, and phospholamban and ryanodine receptor protein levels and phosphorylation become reduced. In FTG, the SR Ca2+ load increases and Ca2+ release following excitation, increases further. An enhanced NCX function in FTG, as reflected by an accelerated relaxation of the caffeine-induced Ca2+ transient, is insufficient to maintain a normal diastolic Ca2+ during high rates of stimulation. Although a high SR Ca2+ release following excitation is maintained, the hypercontractile state is not maintained at high rates of stimulation, and signs of both systolic and diastolic contractile failure appear. Thus, the dilated cardiomyopathy that evolves in this mouse model exhibits signs of both systolic and diastolic failure, but not a deficient SR Ca2+ loading or release, as occurs in some other cardiomyopathic models. PMID- 19125185 TI - Ancestral vascular lumen formation via basal cell surfaces. AB - The cardiovascular system of bilaterians developed from a common ancestor. However, no endothelial cells exist in invertebrates demonstrating that primitive cardiovascular tubes do not require this vertebrate-specific cell type in order to form. This raises the question of how cardiovascular tubes form in invertebrates? Here we discovered that in the invertebrate cephalochordate amphioxus, the basement membranes of endoderm and mesoderm line the lumen of the major vessels, namely aorta and heart. During amphioxus development a laminin containing extracellular matrix (ECM) was found to fill the space between the basal cell surfaces of endoderm and mesoderm along their anterior-posterior (A-P) axes. Blood cells appear in this ECM-filled tubular space, coincident with the development of a vascular lumen. To get insight into the underlying cellular mechanism, we induced vessels in vitro with a cell polarity similar to the vessels of amphioxus. We show that basal cell surfaces can form a vascular lumen filled with ECM, and that phagocytotic blood cells can clear this luminal ECM to generate a patent vascular lumen. Therefore, our experiments suggest a mechanism of blood vessel formation via basal cell surfaces in amphioxus and possibly in other invertebrates that do not have any endothelial cells. In addition, a comparison between amphioxus and mouse shows that endothelial cells physically separate the basement membranes from the vascular lumen, suggesting that endothelial cells create cardiovascular tubes with a cell polarity of epithelial tubes in vertebrates and mammals. PMID- 19125186 TI - Iodine atoms: a new molecular feature for the design of potent transthyretin fibrillogenesis inhibitors. AB - The thyroid hormone and retinol transporter protein known as transthyretin (TTR) is in the origin of one of the 20 or so known amyloid diseases. TTR self assembles as a homotetramer leaving a central hydrophobic channel with two symmetrical binding sites. The aggregation pathway of TTR into amiloid fibrils is not yet well characterized but in vitro binding of thyroid hormones and other small organic molecules to TTR binding channel results in tetramer stabilization which prevents amyloid formation in an extent which is proportional to the binding constant. Up to now, TTR aggregation inhibitors have been designed looking at various structural features of this binding channel others than its ability to host iodine atoms. In the present work, greatly improved inhibitors have been designed and tested by taking into account that thyroid hormones are unique in human biochemistry owing to the presence of multiple iodine atoms in their molecules which are probed to interact with specific halogen binding domains sitting at the TTR binding channel. The new TTR fibrillogenesis inhibitors are based on the diflunisal core structure because diflunisal is a registered salicylate drug with NSAID activity now undergoing clinical trials for TTR amyloid diseases. Biochemical and biophysical evidence confirms that iodine atoms can be an important design feature in the search for candidate drugs for TTR related amyloidosis. PMID- 19125187 TI - Toll-like receptors and cytokines as surrogate biomarkers for evaluating vaginal immune response following microbicide administration. AB - Topical microbicides are intended for frequent use by women in reproductive age. Hence, it is essential to evaluate their impact on mucosal immune function in the vagina. In the present study, we evaluated nisin, a naturally occurring antimicrobial peptide (AMP), for its efficacy as an intravaginal microbicide. Its effect on the vaginal immune function was determined by localizing Toll-like receptors (TLRs-3, 9) and cytokines (IL-4, 6 , 10 and TNF-alpha) in the rabbit cervicovaginal epithelium following intravaginal administration of high dose of nisin gel for 14 consecutive days. The results revealed no alteration in the expression of TLRs and cytokines at both protein and mRNA levels. However, in SDS gel-treated group, the levels were significantly upregulated with the induction of NF-kappaB signalling cascade. Thus, TLRs and cytokines appear as sensitive indicators for screening immunotoxic potential of candidate microbicides. PMID- 19125188 TI - Proinflammatory markers in prediction of posttraumatic psychological symptoms: a prospective cohort study. AB - INTRODUCTION: Posttraumatic psychopathology (PTP) describes the spectrum of conditions that can complicate the recovery from commonly occurring musculoskeletal trauma. There is a clear association with the activation of the hypothalamic-pituitary-adrenal axis (HPAA), and we wished to examine the predictive value of proinflammatory markers of the HPAA and of the GABA, which acts as an inhibitory regulator. METHODS: Levels of proinflammatory markers and GABA were measured in 84 patients who had suffered musculoskeletal injuries requiring hospitalisation. PTP was assessed by the use of the General Health Questionnaire (GHQ) at presentation and again at two- and six-month reviews. RESULTS: Significant psychological disturbance was noted in 39% of patients at two months and falling back to 18% by six months. There was no correlation between any of the markers tested at presentation and PTP at follow-up. DISCUSSION: The HPAA response to trauma and the development of PTP are extremely complex. It is unlikely that a simple blood assay will provide significant predictive information, while incident specific information and patient perception are of more practical use. PMID- 19125189 TI - Temporal dynamics of interferon gamma responses in children evaluated for tuberculosis. AB - BACKGROUND: Development of T-cells based-Interferon gamma (IFNgamma) assays has offered new possibilities for the diagnosis of latent tuberculosis infection (LTBI) and active disease in adults. Few studies have been performed in children, none in France. With reference to the published data on childhood TB epidemiology in the Paris and Ile de France Region, we considered it important to evaluate the performance of IGRA (QuantiFERON TB Gold In Tube(R), QF-TB-IT) in the diagnosis and the follow-up through treatment of LTBI and active TB in a cohort of French children. METHODOLOGY/PRINCIPAL FINDINGS: 131 children were recruited during a prospective and multicentre study (October 2005 and May 2007; Ethical Committee St Louis Hospital, Paris, study number 2005/32). Children were sampled at day 0, 10, 30, 60 (except Healthy Contacts, HC) and 90 for LTBI and HC, and a further day 120, and day 180 for active TB children. Median age was 7.4 years, with 91% of the children BCG vaccinated. LTBI and active TB children undergoing therapy produced significant higher IFNgamma values after 10 days of treatment (p = 0.035). In addition, IFNgamma values were significantly lower at the end of treatment compared to IFNgamma values at day 0, although the number of positive patients was not significantly different between day 0 and end of treatment. CONCLUSIONS/ SIGNIFICANCE: By following quantitative IFNgamma values in each enrolled child with LTBI or active TB and receiving treatment, we were able to detect an increase in the IFNgamma response at day 10 of treatment which might allow the confirmation of a diagnosis. In addition, a decline in IFNgamma values during treatment makes it possible for clinicians to monitor the effect of preventive or curative therapy. PMID- 19125191 TI - Protein palmitoylation regulates osteoblast differentiation through BMP-induced osterix expression. AB - Osteoporosis is one of the most common diseases and can be treated by either anti resorption drugs, anabolic drugs, or both. To search for anabolic drug targets for osteoporosis therapy, it is crucial to understand the biology of bone forming cells, osteoblasts, in terms of their proliferation, differentiation, and function. Here we found that protein palmitoylation participates in signaling pathways that control osterix expression and osteoblast differentiation. Mouse calvarial osteoblasts express most of the 24 palmitoyl transferases, with some being up-regulated during differentiation. Inhibition of protein palmitoylation, with a substrate-analog inhibitor, diminished osteoblast differentiation and mineralization, but not proliferation or survival. The decrease in differentiation capacity is associated with a reduction in osterix, but not Runx2 or Atf4. Inhibition of palmitoyl transferases had little effect in p53(-/-) osteoblasts that show accelerated differentiation due to overexpression of osterix, suggesting that osterix, at least partially, mediated the effect of inhibition of palmitoyl transferases on osteoblast differentiation. BMPs are the major driving force of osteoblast differentiation in the differentiation assays. We found that inhibition of palmitoyl transferases also compromised BMP2-induced osteoblast differentiation through down-regulating osterix induction. However, palmitoyl transferases inhibitor did not inhibit Smad1/5/8 activation. Instead, it compromised the activation of p38 MAPK, which are known positive regulators of osterix expression and differentiation. These results indicate that protein palmitoylation plays an important role in BMP-induced MAPK activation, osterix expression, and osteoblast differentiation. PMID- 19125192 TI - Strong eukaryotic IRESs have weak secondary structure. AB - BACKGROUND: The objective of this work was to investigate the hypothesis that eukaryotic Internal Ribosome Entry Sites (IRES) lack secondary structure and to examine the generality of the hypothesis. METHODOLOGY/PRINCIPAL FINDINGS: IRESs of the yeast and the fruit fly are located in the 5'UTR immediately upstream of the initiation codon. The minimum folding energy (MFE) of 60 nt RNA segments immediately upstream of the initiation codons was calculated as a proxy of secondary structure stability. MFE of the reverse complements of these 60 nt segments was also calculated. The relationship between MFE and empirically determined IRES activity was investigated to test the hypothesis that strong IRES activity is associated with weak secondary structure. We show that IRES activity in the yeast and the fruit fly correlates strongly with the structural stability, with highest IRES activity found in RNA segments that exhibit the weakest secondary structure. CONCLUSIONS: We found that a subset of eukaryotic IRESs exhibits very low secondary structure in the 5'-UTR sequences immediately upstream of the initiation codon. The consistency in results between the yeast and the fruit fly suggests a possible shared mechanism of cap-independent translation initiation that relies on an unstructured RNA segment. PMID- 19125190 TI - Valproate, a mood stabilizer, induces WFS1 expression and modulates its interaction with ER stress protein GRP94. AB - BACKGROUND: Valproate is a standard treatment for bipolar disorder and a first line mood stabilizer. The molecular mechanisms underlying its actions in bipolar disorder are unclear. It has been suggested that the action of valproate is linked to changes in gene expression and induction of endoplasmic reticulum (ER) stress-response proteins. PRINCIPAL FINDINGS: Here we show that valproate modulates the ER stress response through the regulation of WFS1, an important component for mitigating ER stress. Therapeutic concentrations of valproate induce expression of WFS1 mRNA and activate the WFS1 promoter. In addition, WFS1 forms a complex with GRP94, an ER stress-response protein, in which valproate dose-dependently enhances its dissociation from GRP94. CONCLUSIONS: These results suggest that the therapeutic effects of valproate in bipolar disorder may be mediated by WFS1 expression and its dissociation from GRP94. PMID- 19125194 TI - A novel intraurethral device diagnostic index to classify bladder outlet obstruction in men with lower urinary tract symptoms. AB - Objectives. Using a urethral device at the fossa navicularis, bladder pressure during voiding can be estimated by a minimal invasive technique. This study purposes a new diagnostic index for patients with lower urinary tract symptoms (LUTSs). Methods. Fifty one patients presenting with LUTSs were submitted to a conventional urodynamic and a minimal invasive study. The results obtained through the urethral device and invasive classic urodynamics were compared. The existing bladder outlet obstruction index (BOOI) equation that classifies men with LUTSs was modified to allow minimal invasive measurement of isovolumetric bladder pressure in place of detrusor pressure at maximum urine flow. Accuracy of the new equation for classifying obstruction was then tested in this group of men. Results. The modified equation identified men with obstruction with a positive predictive value of 68% and a negative predictive value of 70%, with an overall accuracy of 70%. Conclusions. The proposed equation can accurately classify over 70% of men without resorting to invasive pressure flow studies. We must now evaluate the usefulness of this classification for the surgical treatment of men with LUTSs. PMID- 19125193 TI - IL2RA/CD25 gene polymorphisms: uneven association with multiple sclerosis (MS) and type 1 diabetes (T1D). AB - BACKGROUND: IL-2 receptor (IL2R) alpha is the specific component of the high affinity IL2R system involved in the immune response and in the control of autoimmunity. METHODS AND RESULTS: Here we perform a replication and fine mapping of the IL2RA gene region analyzing 3 SNPs previously associated with multiple sclerosis (MS) and 5 SNPs associated with type 1 diabetes (T1D) in a collection of 798 MS patients and 927 matched Caucasian controls from the south of Spain. We observed association with MS in 6 of 8 SNPs. The rs1570538, at the 3'- UTR extreme of the gene, previously reported to have a weak association with MS, is replicated here (P = 0.032). The most associated T1D SNP (rs41295061) was not associated with MS in the present study. However, the rs35285258, belonging to another independent group of SNPs associated with T1D, showed the maximal association in this study but different risk allele. We replicated the association of only one (rs2104286) of the two IL2RA SNPs identified in the recently performed genome-wide association study of MS. CONCLUSIONS: These findings confirm and extend the association of this gene with MS and reveal a genetic heterogeneity of the associated polymorphisms and risk alleles between MS and T1D suggesting different immunopathological roles of IL2RA in these two diseases. PMID- 19125195 TI - Characterization of a novel polymorphism in PPARG regulatory region associated with type 2 diabetes and diabetic retinopathy in Italy. AB - Peroxisome proliferator-activated receptor gamma polymorphisms have been widely associated with type 2 diabetes, although their role in the pathogenesis of vascular complications is not yet demonstrated. In this study, a cohort of 211 type 2 diabetes, 205 obese, and 254 control individuals was genotyped for Pro12Ala, C1431T, C-2821T polymorphisms, and for a newly identified polymorphism (A-2819G). The above-mentioned polymorphisms were analyzed by gene-specific PCR and direct sequencing of all samples. A significant difference was found for 2819G frequency when patients with type 2 diabetes-particularly diabetic women with the proliferative retinopathy-were compared with healthy control individuals. In conclusion, we identified a novel polymorphism, A-2819G, in PPARG gene, and we found it to be associated with type 2 diabetes and proliferative retinopathy in diabetic females. In the analyzed population, this variant represents a genetic risk factor for developing the diabetic retinopathy, whereas Pro12Ala and C1431T do not. PMID- 19125196 TI - Using discharge abstracts to evaluate a regional perinatal network: assessment of the linkage procedure of anonymous data. AB - To assess the Burgundy perinatal network (18 obstetrical units; 18 500 births per year), discharge abstracts and additional data were collected for all mothers and newborns. In accordance with French law, data were rendered anonymous before statistical analysis, and were linked to patients using a specific procedure. This procedure allowed data concerning each mother to be linked to those for her newborn(s). This study showed that all mothers and newborns were included in the regional database; the data for all mothers were linked to those for their infant(s) in all cases. Additional data (gestational age) were obtained for 99.9% of newborns. PMID- 19125197 TI - Statins Activate Human PPARalpha Promoter and Increase PPARalpha mRNA Expression and Activation in HepG2 Cells. AB - Statins increase peroxisome proliferator-activated receptor alpha (PPARalpha) mRNA expression, but the mechanism of this increased PPARalpha production remains elusive. To examine the regulation of PPARalpha production, we examined the effect of 7 statins (atorvastatin, cerivastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin) on human PPARalpha promoter activity, mRNA expression, nuclear protein levels, and transcriptional activity. The main results are as follows. (1) Majority of statins enhanced PPARalpha promoter activity in a dose-dependent manner in HepG2 cells transfected with the human PPARalpha promoter. This enhancement may be mediated by statin-induced HNF 4alpha. (2) PPARalpha mRNA expression was increased by statin treatment. (3) The PPARalpha levels in nuclear fractions were increased by statin treatment. (4) Simvastatin, pravastatin, and cerivastatin markedly enhanced transcriptional activity in 293T cells cotransfected with acyl-coenzyme A oxidase promoter and PPARalpha/RXRalpha expression vectors. In summary, these data demonstrate that PPARalpha production and activation are upregulated through the PPARalpha promoter activity by statin treatment. PMID- 19125198 TI - Outcome of treatment of anterior vaginal wall prolapse and stress urinary incontinence with transobturator tension-free vaginal mesh (prolift) and concomitant tension-free vaginal tape-obturator. AB - Objective. It is to assess the feasibility, effectiveness, and safety of transobturator tension-free vaginal mesh (Prolift) and concomitant tension-free vaginal tape-obturator (TVT-O) system as a treatment of female anterior vaginal wall prolapse associated with stress urinary incontinence (SUI). Patients and Methods. Between December 2006 and July 2007, 20 patients with anterior genital prolapse and voiding dysfunction were treated with the transobturator tension free vaginal mesh (Prolift) and concomitant tension-free vaginal tape-obturator (TVT-O). Sixteen patients had stress urinary incontinence and 4 patients were considered at risk for development of de novo stress incontinence after the prolapse is repaired. All patients underwent a complete urodynamic assessment. All the patients underwent pelvic examination 4-6 weeks after the operation, and anatomical and functional outcomes were recorded. Results. Twenty cystocoeles were repaired: 6 grade II, 12 grade III, and 2 grade IV. There were no vessel or bladder injuries. Eighteen patients had optimal anatomic results and 2 patients had persistent asymptomatic stage I prolapse. Conclusion. These preliminary results suggest that Prolift system offers a safe and effective treatment for female anterior vaginal wall prolapse. However, a long-term followup is necessary in order to support the good result maintenance. PMID- 19125199 TI - Endourologic management of upper tract transitional cell carcinoma following cystectomy and urinary diversion. AB - Traditionally, nephroureterectomy is the gold standard therapy for upper tract recurrence of transitional cell carcinoma (TCC) following cystectomy and urinary diversion. With advances in endoscopic equipment and improvements in technique, conservative endourologic management via a retrograde or antegrade approach is technically feasible with acceptable outcomes in patients with bilateral disease, solitary renal units, chronic renal insufficiency, or significant medical comorbidities. Contemporary studies have expanded the utility of these techniques to include low-grade, low-volume disease in patients with a normal contralateral kidney. The aim of this report is to review the current outcomes of conservative management for upper tract disease and discuss its application and relevance in patients following cystectomy with lower urinary tract reconstruction. PMID- 19125200 TI - Using pathway signatures as means of identifying similarities among microarray experiments. AB - Widespread use of microarrays has generated large amounts of data, the interrogation of the public microarray repositories, identifying similarities between microarray experiments is now one of the major challenges. Approaches using defined group of genes, such as pathways and cellular networks (pathway analysis), have been proposed to improve the interpretation of microarray experiments. We propose a novel method to compare microarray experiments at the pathway level, this method consists of two steps: first, generate pathway signatures, a set of descriptors recapitulating the biologically meaningful pathways related to some clinical/biological variable of interest, second, use these signatures to interrogate microarray databases. We demonstrate that our approach provides more reliable results than with gene-based approaches. While gene-based approaches tend to suffer from bias generated by the analytical procedures employed, our pathway based method successfully groups together similar samples, independently of the experimental design. The results presented are potentially of great interest to improve the ability to query and compare experiments in public repositories of microarray data. As a matter of fact, this method can be used to retrieve data from public microarray databases and perform comparisons at the pathway level. PMID- 19125201 TI - A description of the origins, design and performance of the TRAITS-SGP Atlantic salmon Salmo salar L. cDNA microarray. AB - The origins, design, fabrication and performance of an Atlantic salmon microarray are described. The microarray comprises 16 950 Atlantic salmon-derived cDNA features, printed in duplicate and mostly sourced from pre-existing expressed sequence tag (EST) collections [SALGENE and salmon genome project (SGP)] but also supplemented with cDNAs from suppression subtractive hybridization libraries and candidate genes involved in immune response, protein catabolism, lipid metabolism and the parr-smolt transformation. A preliminary analysis of a dietary lipid experiment identified a number of genes known to be involved in lipid metabolism. Significant fold change differences (as low as 1.2x) were apparent from the microarray analysis and were confirmed by quantitative real-time polymerase chain reaction analysis. The study also highlighted the potential for obtaining artefactual expression patterns as a result of cross-hybridization of similar transcripts. Examination of the robustness and sensitivity of the experimental design employed demonstrated the greater importance of biological replication over technical (dye flip) replication for identification of a limited number of key genes in the studied system. The TRAITS (TRanscriptome Analysis of Important Traits of Salmon)-salmon genome project microarray has been proven, in a number of studies, to be a powerful tool for the study of key traits of Atlantic salmon biology. It is now available for use by researchers in the wider scientific community. PMID- 19125202 TI - Influenza a virus induces an immediate cytotoxic activity in all major subsets of peripheral blood mononuclear cells. AB - BACKGROUND: A replication defective influenza A vaccine virus (delNS1 virus) was developed. Its attenuation is due to potent stimulation of the innate immune system by the virus. Since the innate immune system can also target cancer cells, we reasoned that delNS1 virus induced immune-stimulation should also lead to the induction of innate cytotoxic effects towards cancer cells. METHODOLOGY/PRINCIPAL FINDINGS: Peripheral blood mononuclear cells (PBMCs), isolated CD56+, CD3+, CD14+ and CD19+ subsets and different combinations of the above subsets were stimulated by delNS1, wild type (wt) virus or heat inactivated virus and co-cultured with tumor cell lines in the presence or absence of antibodies against the interferon system. Stimulation of PBMCs by the delNS1 virus effectively induced cytotoxicity against different cancer cell lines. Surprisingly, virus induced cytotoxicity was exerted by all major subtypes of PBMCs including CD56+, CD3+, CD14+ and CD19+ cells. Virus induced cytotoxicity in CD3+, CD14+ and CD19+ cells was dependent on virus replication, whereas virus induced cytotoxicity in CD56+ cells was only dependent on the binding of the virus. Virus induced cytotoxicity of isolated cell cultures of CD14+, CD19+ or CD56+ cells could be partially blocked by antibodies against type I and type II (IFN) interferon. In contrast, virus induced cytotoxicity in the complete PBMC preparation could not be inhibited by blocking type I or type II IFN, indicating a redundant system of activation in whole blood. CONCLUSIONS/SIGNIFICANCE: Our data suggest that apart from their well known specialized functions all main subsets of peripheral blood cells also initially exert a cytotoxic effect upon virus stimulation. This closely links the innate immune system to the adaptive immune response and renders delNS1 virus a potential therapeutic tool for viro-immunotherapy of cancer. PMID- 19125203 TI - Antiretroviral-related adipocyte dysfunction and lipodystrophy in HIV-infected patients: Alteration of the PPARgamma-dependent pathways. AB - Lipodystrophy and metabolic alterations are major complications of antiretroviral therapy in HIV-infected patients. In vitro studies using cultured murine and human adipocytes revealed that some protease inhibitors (PIs) and nucleoside reverse transcriptase inhibitors (NRTIs) were implicated to a different extent in adipose cell dysfunction and that a chronic incubation with some PIs decreased mRNA and protein expression of PPARgamma. Defective lamin A maturation linked to PI inhibitory activity could impede the nuclear translocation of SREBP1c, therefore, reducing PPARgamma expression. Adipose cell function was partially restored by the PPARgamma agonists, thiazolidinediones. Adverse effects of PIs and NRTIs have also been reported in macrophages, a cell type that coexists with, and modulates, adipocyte function in fat tissue. In HIV-infected patients under ART, a decreased expression of PPARgamma and of PPARgamma-related genes was observed in adipose tissue, these anomalies being more severe in patients with ART-induced lipoatrophy. Altered PPARgamma expression was reversed in patients stopping PIs. Treatment of patients with agonists of PPARgamma could improve, at least partially, the subcutaneous lipoatrophy. These data indicate that decreased PPARgamma expression and PPARgamma-related function, resulting from ART-induced adipose tissue toxicity, play a central role in HIV-related lipoatrophy and metabolic consequences. PMID- 19125204 TI - Proinflammation and hypertension: a population-based study. AB - There is evidence that proinflammation may be linked to the development of hypertension (HT). We examined the association of both the interleukin-1 beta (IL 1beta) and the interleukin 1-receptor antagonist (IL-1ra) with future blood pressure (BP) and HT occurrence (BP >or= 140/90 mmHg, or antihypertensive drug) in a population-based prospective study. Our study consisted of 396 (147 men and 249 women) middle-aged, baseline apparently healthy, normotensive subjects participating in a 6.5-year follow-up study. Subjects with high-sensitivity CRP (hs-CRP) < 10 mg/L were excluded at the initial visit. At follow-up, the occurrence of HT was 32%. The levels of baseline IL-1beta and IL-1ra were significantly higher for subjects who developed HT during the follow-up than for those who did not (IL-1beta; 0.67 +/- 0.62 pg/mL versus 0.56 +/- 0.32 pg/mL, P = .020 and IL-1ra; 184 +/- 132 pg/mL versus 154 +/- 89 pg/mL, P = .007). After adjustments for age, follow-up time, sex, baseline systolic BP, and BMI, our results confirm a statistically significant (P = .036) linear association between the quartiles of IL-1beta and change of systolic BP during the study. After adjustments for age, follow-up time, sex, and BMI, our results also show a linear association between incident HT and the quartiles of IL-1ra. (P = .026). These results provide evidence that proinflammation may precede BP elevation and HT. PMID- 19125205 TI - Will patients benefit from regionalization of gynecologic cancer care? AB - OBJECTIVE: Patient chances for cure and palliation for a variety of malignancies may be greatly affected by the care provided by a treating hospital. We sought to determine the effect of volume and teaching status on patient outcomes for five gynecologic malignancies: endometrial, cervical, ovarian and vulvar carcinoma and uterine sarcoma. METHODS: The Florida Cancer Data System dataset was queried for all patients undergoing treatment for gynecologic cancers from 1990-2000. RESULTS: Overall, 48,981 patients with gynecologic malignancies were identified. Endometrial tumors were the most common, representing 43.2% of the entire cohort, followed by ovarian cancer (30.9%), cervical cancer (20.8%), vulvar cancer (4.6%), and uterine sarcoma (0.5%). By univariate analysis, although patients treated at high volume centers (HVC) were significantly younger, they benefited from an improved short-term (30-day and/or 90-day) survival for cervical, ovarian and endometrial cancers. Multivariate analysis (MVA), however, failed to demonstrate significant survival benefit for gynecologic cancer patients treated at teaching facilities (TF) or HVC. Significant prognostic factors at presentation by MVA were age over 65 (HR = 2.6, p<0.01), African-American race (HR = 1.36, p<0.01), and advanced stage (regional HR = 2.08, p<0.01; advanced HR = 3.82, p<0.01, respectively). Surgery and use of chemotherapy were each significantly associated with improved survival. CONCLUSION: No difference in patient survival was observed for any gynecologic malignancy based upon treating hospital teaching or volume status. Although instances of improved outcomes may occur, overall further regionalization would not appear to significantly improve patient survival. PMID- 19125206 TI - Sex cord-gonadal stromal tumor of the rete testis. AB - A 34-year-old tetraplegic patient with suppurative epididymitis was found on follow-up examination and ultrasonography to have a testicular mass. The radical orchiectomy specimen contained an undifferentiated spindled sex cord-stromal tumor arising in the rete testis. Testicular sex cord-stromal tumors are far less common than germ cell neoplasms and are usually benign. The close relationship between sex cords and ductules of the rete testis during development provides the opportunity for these uncommon tumors to arise anatomically within the rete tesis. This undifferentiated sex cord-stromal tumor, occurring in a previously unreported location, is an example of an unusual lesion mimicking an intratesticular malignant neoplasm. PMID- 19125207 TI - Postpartum invasive group A streptococcal disease in the modern era. AB - To describe the clinical features of individuals hospitalized for postpartum invasive group A Streptococcus (GAS) infection, a retrospective, population-based study of hospitalized patients in the state of Florida was conducted. Cases of postpartum invasive GAS infection (occurring within 42 days of delivery) were compared to women with other manifestations of invasive GAS disease with respect to their age at the time of admission. Four cases of postpartum invasive GAS infection were detected in this population, yielding a prevalence of 1.6% (4/257) of postpartum disease in this invasive GAS infection database. Patients presented a median of 4 days (mean of 9 days) after delivery with signs and symptoms of infection. Three cases were complicated by bacteremia and one patient had streptococcal toxic shock syndrome. Each patient received multiple antibiotics and survived. No patients received intravenous immunoglobulin. For comparison, a secondary retrospective investigation of a large hospital discharge dataset obtained from the Florida Agency for Health Care Administration was assessed for patients with puerperal GAS infections. This method yielded an additional three cases, whose clinical and demographic characteristics were summarized. These data highlight that postpartum invasive GAS infection continues to complicate pregnancy, though the frequency has decreased markedly over the past century. PMID- 19125208 TI - Has the time come for clinical trials on the antidepressant effect of vitamin D? PMID- 19125210 TI - Neural synchrony in patients with a first episode of schizophrenia: tracking relations with grey matter and symptom profile. AB - BACKGROUND: Although schizophrenia has been characterized by disruptions to neural synchrony, it remains unknown whether these disturbances are related to symptoms and loss of grey matter. We examined relations between 40 Hz Gamma band synchrony and grey matter in patients with schizophrenia at first episode and after 2.5 years. METHODS: From an initial recruitment of 35 medicated patients with a first episode of schizophrenia, 25 patients completed clinical and oddball task-elicited Gamma synchrony within 3 months of health service contact and again after 2.5 years, 23 completed magnetic resonance imaging (MRI) at these time points, and 13 completed all sessions. We compared patients with 35 matched healthy controls. We identified early (0-150 ms) and late (250-500 ms) peaks in Gamma synchrony locked to oddball targets, and we analyzed MRI data using voxel based morphometry. We evaluated group and test-retest differences using repeated measures analyses of variance. RESULTS: Compared with controls, at first contact, patients with a first episode of schizophrenia showed a disruption to the laterality of early Gamma synchrony and global reduction in late Gamma synchrony, with a corresponding loss of fronto-temporal-parietal grey matter. Gamma synchrony was increased at follow-up among patients with a first episode of schizophrenia. It related negatively to further loss of grey matter, but positively to improvement in reality distortion symptoms. These relations could not be explained by medication dose. LIMITATIONS: Our study did not include unmedicated patients or normative follow-up testing. CONCLUSION: Gamma synchrony may track the progression of schizophrenia from first episode. An increase in Gamma synchrony over time might reflect an attempt to adapt to a progressive loss of cortical grey matter and associated changes in cognitive and emotional function. PMID- 19125209 TI - Cascading effects of stressors and inflammatory immune system activation: implications for major depressive disorder. AB - Activation of the inflammatory immune system provokes numerous neuroendocrine and neurotransmitter changes, many of which are similar to those provoked by physical or psychological stressors. These findings, among others, have led to the suggestion that the brain translates immune activation much as if it were a stressor. In this review, I provide synopses of the effects of traditional stressors on the release of corticotropin-releasing hormones at hypothalamic and extrahypothalamic sites, variations of serotonin and its receptors and changes of brain-derived neurotrophic factor (BDNF). These effects are similar to those elicited by activation of the inflammatory immune system, particularly the impact of the immune-signalling molecules interleukin-1 beta, interleukin-6, tumour necrosis factor-alpha and interferon-alpha on neuroendocrine, neurotransmitter and BDNF function. In addition, it is reported that stressors and cytokines may synergistically influence biological and behavioural processes and that these treatments may have long-term ramifications through the sensitization of processes associated with stress responses. Finally, I present an overview of the depressogenic actions of these cytokines in rodent models and in humans, and I provide provisional suggestions (and caveats) about the mechanisms by which cytokines and stressors might culminate in major depressive disorder. PMID- 19125211 TI - Genotype over-diagnosis in amygdala responsiveness: affective processing in social anxiety disorder. AB - BACKGROUND: Although the amygdala is thought to be a crucial brain region for negative affect, neuroimaging studies do not always show enhanced amygdala response to aversive stimuli in patients with anxiety disorders. Serotonin (5-HT) related genotypes may contribute to interindividual variability in amygdala responsiveness. The short (s) allele of the 5-HT transporter linked polymorphic region (5-HTTLPR) and the T variant of the G-703T polymorphism in the tryptophan hydroxylase-2 (TPH2) gene have previously been associated with amygdala hyperresponsivity to negative faces in healthy controls. We investigated the influence of these polymorphisms on amygdala responsiveness to angry faces in patients with social anxiety disorder (SAD) compared with healthy controls. METHODS: We used positron emission tomography with oxygen 15-labelled water to assess regional cerebral blood flow in 34 patients with SAD and 18 controls who viewed photographs of angry and neutral faces presented in counterbalanced order. We genotyped all participants with respect to the 5-HTTLPR and TPH2 polymorphisms. RESULTS: Patients with SAD and controls had increased left amygdala activation in response to angry compared with neutral faces. Genotype but not diagnosis explained a significant portion of the variance in amygdala responsiveness, the response being more pronounced in carriers of s and/or T alleles. LIMITATIONS: Our analyses were limited owing to the small sample and the fact that we were unable to match participants on genotype before enrollment. In addition, other imaging techniques not used in our study may have revealed additional effects of emotional stimuli. CONCLUSION: Amygdala responsiveness to angry faces was more strongly related to serotonergic polymorphisms than to diagnosis of SAD. Emotion activation studies comparing amygdala excitability in patient and control groups could benefit from taking variation in 5-HT-related genes into account. PMID- 19125212 TI - A meta-analysis examining clinical predictors of hippocampal volume in patients with major depressive disorder. AB - BACKGROUND: Some, although not all, studies report small hippocampal volume in patients with major depressive disorder (MDD) relative to healthy controls. Here, we explore the contribution of key demographic and clinical variables to this difference. METHODS: We used meta-analytic techniques to provide an updated analysis of data from 32 magnetic resonance imaging studies of hippocampal volume in patients with MDD. RESULTS: Our analysis confirmed the difference in hippocampal volume, but only among patients with MDD whose duration of illness was longer than 2 years or who had more than 1 disease episode. We found no such effect in studies that included patients who did not fit these criteria. The effect was limited to children and middle-aged or older adults. Analyzed collectively, studies including young adult patients showed equivalent hippocampal volumes across MDD patients and controls, a result that may be attributable to a reduced burden of illness in this population. Age at onset of disease, severity of depression at the time of scanning, sex and slice thickness did not contribute to differences in hippocampal volume between patients with MDD and controls. LIMITATIONS: The small size of many of the clinical and demographic subgroups may have limited statistical power to detect between-group differences. CONCLUSION: Although all studies were cross-sectional, our results suggest that hippocampal volume reductions generally occur after disease onset in patients with MDD. These findings have implications for the timing of clinical interventions aimed at reducing the impact of MDD on neuronal structure and function. PMID- 19125215 TI - The 31st annual meeting of the Canadian College of Neuropsychopharmacology: Toronto, Ontario, June 6-9, 2008. PMID- 19125213 TI - A new wave in the genetics of psychiatric disorders: the copy number variant tsunami. PMID- 19125214 TI - High-frequency repetitive transcranial magnetic stimulation to the cerebellum and implicit processing of happy facial expressions. AB - BACKGROUND: Previous research has demonstrated that the cerebellum is involved in emotive and cognitive processes. Furthermore, recent findings suggest high frequency repetitive transcranial magnetic stimulation (rTMS) to the cerebellum has mood-improving properties. We sought to further explore the effects of cerebellar high-frequency rTMS on implicit processing of emotional stimuli and mood. METHODS: In a double-blind, crossover study, 15 healthy volunteers received 15 minutes of 20 Hz (5 s on, 5 s off) rTMS over the medial cerebellum, occipital cortex or sham in a randomized counterbalanced order on 3 consecutive days. A masked emotional faces response task measured implicit emotional processing of happy, fearful and neutral facial expressions. We used positive and negative affect scales to evaluate rTMS-related changes in mood. RESULTS: High-frequency rTMS over the cerebellum was associated with significant increases in masked emotional responses to happy facial expressions only. We observed no changes in consciously experienced mood. LIMITATIONS: Although the sham rTMS served as our baseline measurement, additional pre-rTMS data showing that reaction time increases immediately after cerebellar rTMS would have made our results more compelling. CONCLUSION: The results replicate and extend previous findings by establishing a direct relation between the cerebellum and emotive information processing. The parallel between the present effects of high-frequency cerebellar rTMS and short-term antidepressant therapy regarding the change in implicit processing of positive stimuli in the absence of mood changes is notable and warrants further research. PMID- 19125216 TI - Optimal use of antidepressants: when to act? PMID- 19125217 TI - The contribution of transposable elements to expressed coding sequence in Arabidopsis thaliana. AB - The goal of this study was to assess the extent to which transposable elements (TEs) have contributed to protein-coding regions in Arabidopsis thaliana. To do this, we first characterized the extent of chimeric TE-gene constructs. We compared a genome-wide TE database to genomic sequences, annotated coding regions, and EST data. The comparison revealed that 7.8% of expressed genes contained a region with close similarity to a known TE sequence. Some groups of TEs, such as helitrons, were underrepresented in exons relative to their genome wide distribution; in contrast, Copia-like and En/Spm-like sequences were overrepresented in exons. These 7.8% percent of genes were enriched for some GO based functions, particularly kinase activity, and lacking in other functions, notably structural molecule activity. We also examined gene family evolution for these genes. Gene family information helped clarify whether the sequence similarity between TE and gene was due to a TE contributing to the gene or, instead, the TE co-opting a portion of the gene. Most (66%) of these genes were not easily assigned to a gene family, and for these we could not infer the direction of the relationship between TE and gene. For the remainder, where appropriate, we built phylogenetic trees to infer the direction of the TE-gene relationship by parsimony. By this method, we verified examples where TEs contributed to expressed proteins. Our results are undoubtedly conservative but suggest that TEs may have contributed small protein segments to as many as 1.2% of all expressed, annotated A. thaliana genes. PMID- 19125218 TI - Necessity of screening water chestnuts for microcystins after cyanobacterial blooms break out. AB - Water chestnut is one of the most popular vegetables in Asian countries that grows in shallow water. Eighteen water chestnut samples were collected from Lake Tai and six samples were bought at markets in Wuxi, China, in October 2007. Extraction solution of water chestnut was cleaned up with a solid phase extraction column and immunoaffinity chromatography cartridges, then the microcystin (MC) level was detected by indirect competitive enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-mass spectrometry (LC-MS). The results of ELISA showed that there were six samples collected from Lake Tai which contained MCs; the highest level of total MCs was 7.02 ng/g. The results of LC-MS confirmed that MC-LR and MC-RR were present in five samples. The highest level of MC-LR was 1.02 ng/g and that of MC-RR was 4.44 ng/g. Heavy cyanobacterial blooms had occurred, and MCs were detected in water at the points in Lake Tai where MCs occurred in water chestnuts collected in 2007. MCs were not detected in the six samples bought at Wuxi markets. The results suggest that MCs can accumulate in water chestnuts, which is a potential hazard for human health. PMID- 19125219 TI - Mycoviruses are common among different species of endophytic fungi of grasses. AB - A survey of mycoviruses was made in a collection of 103 isolates belonging to 53 different species of endophytic fungi of grasses. Double-stranded RNA (dsRNA) elements were detected in isolates of 12 of the species analyzed. The banding characteristics and sizes of some of the dsRNA elements suggest that they might belong to previously described mycovirus families. The observed incidence (22.6%) indicates that the presence of mycoviruses could be common among species of this group of ubiquitous fungi. PMID- 19125220 TI - Purification and biochemical characterization of a native invertase from the hydrogen-producing Thermotoga neapolitana (DSM 4359). AB - This is the first report describing the purification and enzymatic properties of a native invertase (beta-D-fructosidase) in Thermotogales. The invertase of the hydrogen-producing thermophilic bacterium Thermotoga neapolitana DSM 4359 (hereby named Tni) was a monomer of about 47 kDa having an amino acid sequence quite different from other invertases studied up to now. Its properties and substrates specificity let us classify this protein as a solute-binding protein with invertase activity. Tni was specific for the fructose moiety and the enzyme released fructose from sucrose and raffinose and the fructose polymer inulin was hydrolyzed in an endo-type fashion. Tni had an optimum temperature of 85 degrees C at pH 6.0. At temperatures of 80-85 degrees C, the enzyme retained at least 50% of its initial activity during a 6 h preincubation period. Tni had a K(m) and k(cat)/K(m) values (at 85 degrees C and pH 6.0) of about 14 mM and 5.2 x 10(8) M( 1) s(-1), respectively. PMID- 19125221 TI - SUN-domain and KASH-domain proteins during development, meiosis and disease. AB - SUN-domain proteins interact directly with KASH-domain proteins to form protein complexes that connect the nucleus to every major cytoskeleton network. SUN-KASH protein complexes are also required for attaching centrosomes to the nuclear periphery and for alignment of homologous chromosomes, their pairing and recombination in meiosis. Other functions that require SUN-domain proteins include the regulation of apoptosis and maturation and survival of the germline. Laminopathic diseases affect the distribution of the SUN-KASH complexes, and mutations in KASH-domain proteins can cause Emery Dreifuss muscular dystrophy and recessive cerebellar ataxia. This review describes our current knowledge of the role of SUN-KASH domain protein complexes during development, meiosis and disease. PMID- 19125222 TI - Disposition of temozolomide in a patient with glioblastoma multiforme after gastric bypass surgery. AB - Temozolomide, used for anaplastic gliomas and glioblastoma multiforme, is an oral drug that is stable under acidic, but labile under neutral and basic conditions. Although the bioavailability of temozolomide is approximately 100%, pathology or anatomical changes of the gastrointestinal tract may adversely affect absorption, and consequently therapeutic response. HPLC-UV was used to evaluate temozolomide plasma pharmacokinetics in a patient with unresponsive glioblastoma multiforme who had previously undergone gastric bypass as part of a weight-loss strategy. Temozolomide plasma pharmacokinetics were comparable to values reported for patients with normal gastrointestinal anatomy. These data imply that progression of disease in this patient was not due to inadequate temozolomide concentrations. Physicians need to become aware of the rapidly increasing population of patients who have had a gastric bypass and require oral therapy, of which our case is representative. The effect of gastric bypass on pharmacokinetics will need to be evaluated on a drug-by-drug basis. PMID- 19125223 TI - Induction studies with Escherichia coli expressing recombinant interleukin-13 using multi-parameter flow cytometry. AB - The expression of interleukin-13 (IL13) following induction with IPTG in Escherichia coli results in metabolic changes as indicated by multi-parameter flow cytometry and traditional methods of fermentation profiling (O2 uptake rate, CO2 evolution rate and optical density measurements). Induction early in the rapid growth phase was optimal although this led to lower overall biomass concentrations and lower maximum specific growth rates. In contrast, induction in the mid-rapid growth phase was the most detrimental to cell quality as measured by cytoplamsic membrane depolarisation. PMID- 19125224 TI - A novel carbonyl reductase from Pichia stipitis for the production of ethyl (S)-4 chloro-3-hydroxybutanoate. AB - An NADPH-dependent carbonyl reductase (PsCR) gene from Pichia stipitis was cloned. It contains an open reading frame of 849 bp encoding 283 amino acids whose sequence had less than 60% identity to known reductases that produce ethyl (S)-4-chloro-3-hydroxybutanoates (S-CHBE). When expressed in Escherichia coli, the recombinant PsCR exhibited an activity of 27 U/mg using ethyl 4-chloro-3 oxobutanoate (COBE) as a substrate. Reduction of COBE to (S)-CHBE by transformants in an aqueous mono-phase system for 18 h, gave a molar yield of 94% and an optical purity of the (S)-isomer of more than 99% enantiomeric excess. PMID- 19125225 TI - Introduction of a stress-responsive gene, yggG, enhances the yield of L phenylalanine with decreased acetic acid production in a recombinant Escherichia coli. AB - A stress-responsive gene, yggG, was introduced into an L-phenylalanine producer, Escherichia coli AJ12741. In shake-flask culture, the yggG-containing recombinant strain (named AJ12741/pHYGG) produced 6.4 g L-phenylalanine l(-1) at the end of culture and its yield on glucose was 0.16 g L-phenylalanine g glucose(-1). These values are much higher than those of the original AJ12741 strain (3.7 g L phenylalanine l(-1) and 0.09 g L-phenylalanine g glucose(-1), respectively). On the other hand, AJ12741/pHYGG strain produced only 4.5 g acetic acid l(-1) and its yield on glucose was about a half of that of the AJ12741 culture. Analysis of gene expression revealed that in late growth phase, the expression levels of genes involved in acetic acid production (pta, ackA, and poxB) were relatively low in AJ12741/pHYGG cells. In particular, the level of poxB expression in AJ12741/pHYGG strains was one-seventh of that of the original strain. These results suggest that the formation of a bottleneck for acetic acid production brings about a metabolic flow favorable to L-phenylalanine synthesis in the recombinant strain over-expressing the yggG gene. PMID- 19125226 TI - Characterization of Capsicum annuum recombinant alpha- and beta-tubulin. AB - There are several conditions which might modulate polymerization to produce polymers having normal lattice structure. In the absence of 1 mM MgCl(2) the assembly was reduced by 36% in Capsicum annuum tubulin (CAnm tubulin). There was no significant difference in the final assembly formation in the presence of 5% to 10% glycerol. However, nucleation rate was slow and apparent study state was achieved lately in the presence of 10% glycerol. Taxol at 100 microM concentration increased 23% tubulin assembly. One millimolar CaCl(2), >or=1% dimethyl sulfoxide (DMSO) and physiologically low temperature reduced CAnm tubulin assembly. A value of 0.089 mg/ml was obtained as critical concentration for polymerization. Benomyl significantly reduced the number of cysteine residues accessible to 5,5'-dithiobis-(2-nitrobenzoic acid); there were 4.77 +/- 0.21 and 3.49 +/- 0.35 residues accessible per tubulin dimer in the presence of 50 and 100 microM benomyl respectively. PMID- 19125227 TI - Statistical optimization of recycled-paper enzymatic hydrolysis for simultaneous saccharification and fermentation via central composite design. AB - A central composite design of the response surface methodology (RSM) was employed to study the effects of temperature, enzyme concentration, and stirring rate on recycled-paper enzymatic hydrolysis. Among the three variables, temperature and enzyme concentration significantly affected the conversion efficiency of substrate, whereas stirring rate was not effective. A quadratic polynomial equation was obtained for enzymatic hydrolysis by multiple regression analysis using RSM. The results of validation experiments were coincident with the predicted model. The optimum conditions for enzymatic hydrolysis were temperature, enzyme concentration, and stirring rate of 43.1 degrees C, 20 FPU g( 1) substrate, and 145 rpm, respectively. In the subsequent simultaneous saccharification and fermentation (SSF) experiment under the optimum conditions, the highest 28.7 g ethanol l(-1) was reached in the fed-batch SSF when 5% (w/v) substrate concentration was used initially, and another 5% added after 12 h fermentation. This ethanol output corresponded to 77.7% of the theoretical yield based on the glucose content in the raw material. PMID- 19125228 TI - Pretreatment of rice straw by a hot-compressed water process for enzymatic hydrolysis. AB - Hot-compressed water (HCW) is among several cost-effective pretreatment processes of lignocellulosic biomass for enzymatic hydrolysis. The present work investigated the characteristics of HCW pretreatment of rice straw including sugar production and inhibitor formation in the liquid fraction and enzymatic hydrolysis of pretreated material. Pretreatment was carried out at a temperature ranging from 140 to 240 degrees C for 10 or 30 min. Soluble oligosaccharides were found to constitute almost all the components of total sugars in the liquid fraction. The maximal production of total glucose at 180 degrees C and below accounted for 4.4-4.9% of glucan in raw material. Total xylose production peaked at 180 degrees C, accounting for 43.3% of xylan in raw material for 10-min pretreatment and 29.8% for 30-min pretreatment. The production of acetic acid increased at higher temperatures and longer treatment time, indicating more significant disruption of lignocellulosic structure, and furfural production achieved the maximum (2.8 mg/ml) at 200 degrees C for both 10-min and 30-min processes. The glucose yield by enzymatic hydrolysis of pretreated rice straw was no less than 85% at 180 degrees C and above for 30-min pretreatment and at 200 degrees C and above for 10-min pretreatment. Considering sugar recovery, inhibitor formation, and process severity, it is recommended that a temperature of 180 degrees C for a time of 30 min can be the most efficient process for HCW pretreatment of rice straw. PMID- 19125229 TI - Alternative splicing in the synaptic protein interaction site of rat Ca(v)2.2 (alpha (1B)) calcium channels: changes induced by chronic inflammatory pain. AB - Voltage-gated N-type Ca(2+) channels play a central role in regulation of neurotransmitter release. The II-III linker of N-type calcium channel alpha(1B) subunit (exons 17-21 in rats) contains synaptic protein interaction (synprint) site, which plays a pivotal role in efficient neurotransmission. Using RT-PCR analysis of rat hippocampus and lumbar spinal cord transcripts, the expressions of Ca(v)2.2 exon 18a and a novel Ca(v)2.2 splice variant, which were detected in this study, have been investigated, and they showed tissue-specific pattern. The new variant contains a large deletion in II-III linker that produces a stop codon, which is predicted to produce a two-domain-truncated channel. To study the effect of inflammatory pain on the expression pattern of these variants, animals were treated with intraplantar formalin, and the amount of splice variants in lumbar spinal cord was measured. The results showed that chronic inflammatory pain increases the Ca(v)2.2 mRNA levels lacking exon 18a and decreases the amount of full-length variants, which do not have any deletion. Determining different Ca(v)2.2 splice variants in rat nervous system and the impact of inflammatory pain on the splicing pattern suggest a possible regulatory role for calcium channel alternative splicing. PMID- 19125230 TI - Chilblain lupus erythematosus-a review of literature. PMID- 19125231 TI - Shedding light on microcirculation? PMID- 19125233 TI - Case-mix-adjusted length of stay and mortality in 23 Finnish ICUs. AB - OBJECTIVES: To create a tool for benchmarking intensive care units (ICUs) with respect to case-mix adjusted length of stay (LOS) and to study the association between clinical and economic measures of ICU performance. DESIGN: Observational cohort study. SETTING: Twenty-three ICUs in Finland. PATIENTS: A total of 80,854 consecutive ICU admissions during 2000-2005, of which 63,304 met the inclusion criteria. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Linear regression was used to create a model that predicted ICU LOS. Simplified Acute Physiology Score (SAPS) II, age, disease categories according to Acute Physiology and Chronic Health Evaluation III, single highest Therapeutic Intervention Scoring System score collected during the ICU stay and presence of other ICUs in the hospital were included in the model. Probabilities of hospital death were calculated using SAPS II, age, and disease categories as covariates. In the validation sample, the created model accounted for 28% of variation in ICU LOS across individual admissions and 64% across ICUs. The expected ICU LOS was 2.53 +/- 2.24 days and the observed ICU LOS was 3.29 +/- 5.37 days, P < 0.001. There was no association between the mean observed - mean expected ICU LOS and standardized mortality ratios of the ICUs (Spearman correlation 0.091, P = 0.680). CONCLUSIONS: We developed a tool for the assessment of resource use in a large nationwide ICU database. It seems that there is no association between clinical and economic quality indicators. PMID- 19125232 TI - Year in review in Intensive Care Medicine, 2008: II. Experimental, acute respiratory failure and ARDS, mechanical ventilation and endotracheal intubation. PMID- 19125234 TI - Validation of the urine column measurement as an estimation of the intra abdominal pressure. AB - OBJECTIVE: To evaluate the efficacy of the urine column (UC) measurement compared to the intra-vesicular pressure (IVP) measurement as an estimation of intra abdominal pressure (IAP) in patients with IAP up to 30 mmHg. METHODS: Fifteen patients undergoing a laparoscopic cholecystectomy were studied. All patients were catheterized. IVP measurements were performed using a pressure transducer connected to the culture aspiration port. UC measurements were done by holding up the tubing against a measuring rod. The symphysis pubis was used as the zero reference. IAP was raised from 0 to 30 mmHg using increments of 5 mmHg, during which first the IVP and then UC measurement series were recorded end-expiratory. Fifty and 100 ml of saline were used as a priming volume. RESULTS: The IVP and UC measurements showed a significant correlation with IAP. Comparing IVP and UC showed a correlation of 0.91 (p < 0.001) for 50 ml and 0.87 (p < 0.001) for 100 ml of saline as a priming volume. Using 50 ml of saline, UC was 0.68 mmHg higher than IVP (95% CI -7.21 to +5.85 mmHg). For 100 ml of saline, UC was 1.23 mmHg lower than IVP (95% CI -7.41 to +9.87 mmHg). CONCLUSION: UC measurement shows significant correlation to IVP measurement as an estimation of the IAP. Further study needs to be done to conclude whether UC measurement is a reliable clinical alternative to IVP measurement. PMID- 19125236 TI - Novice drivers' performance after different alcohol dosages and placebo in the divided-attention steering simulator (DASS). AB - RATIONALE: The divided-attention steering simulator (DASS) is designed to measure lane-keeping (i.e., a tracking task using a steering wheel) while performing a secondary visual task (responding to digits that appear in the corners of the computer screen). Some studies have already used the DASS, but the magnitude of impairment is difficult to interpret because reference values are lacking. OBJECTIVE: To examine the magnitude of impairment after administration of four different dosages of alcohol and placebo. MATERIALS AND METHODS: Thirty-two healthy young adults participated in this randomized, single-blind crossover trial. Subjects received alcohol to gain a blood alcohol concentration (BAC) of 0.02%, 0.05%, 0.08%, and 0.10% or alcohol-placebo. Sixteen subjects performed a 30-min test in DASS (dual-task condition). Outcome measures were steering error, reaction time, and percentage of errors. Sixteen other subjects performed the test without performing the secondary peripheral task (single-task condition). RESULTS: Twenty-eight subjects (novice drivers; drivers' license up to 5 years) were included in the analyses. Dose-dependent impairment was found in both the single-task condition (F ((4,11)) = 10.86, p < 0.001) and the dual-task condition (F ((4,9)) = 5.58, p < 0.015). Performance at all BAC levels differed significantly (p < 0.05) from alcohol-placebo, except BAC 0.02%. With increasing BAC levels, subjects made more errors and reacted slower on the peripheral visual search task, but these effects did not reach significance. CONCLUSION: With increasing BAC, dose-dependent impairment was found. The DASS seems to be a suitable divided-attention task that is useful in psychopharmacological research and training of novice drivers. PMID- 19125237 TI - A preclinical model of binge eating elicited by yo-yo dieting and stressful exposure to food: effect of sibutramine, fluoxetine, topiramate, and midazolam. AB - RATIONALE: Preclinical models are needed to investigate the neurobiology and psychobiology of binge eating and to identify innovative pharmacotherapeutic strategies. OBJECTIVES: A modification of the model based on the combination of cyclic caloric restrictions and acute stress was developed to further increase its face validity and reliability and, for the first time, to assess its predictive value. MATERIALS AND METHODS: Four groups of female rats were employed: group 1 was normally fed and not stressed on the test day (25th); group 2 was fed normally but was exposed to an acute stress on day 25; group 3 was exposed to three cycles (4 days 66% of chow intake + 4 days food ad libitum) of yo-yo dieting but not stressed; and group 4 was exposed to cyclic yo-yo dieting and then stressed. All groups were fed highly palatable food (HPF) for 2 h on days 5-6 and 13-14. Acute stress was elicited by exposing rats to HPF, but preventing them from access to it for 15 min. RESULTS: The combination of cyclic food restriction and stressful exposure to food markedly increased HPF intake. Sibutramine and fluoxetine inhibited food intake in all conditions. Topiramate selectively inhibited compulsive HPF intake in rats submitted to caloric restriction and stress. Midazolam increased HPF intake. CONCLUSIONS: Pharmacological results suggest that this model, in addition to face validity as an isomorphic model of human binge eating, is endowed with good predictive validity. PMID- 19125235 TI - Increased operant responding for ethanol in male C57BL/6J mice: specific regulation by the ERK1/2, but not JNK, MAP kinase pathway. AB - RATIONALE: Extracellular signal-regulated protein kinase (ERK(1/2)) is a member of the mitogen-activated protein kinase (MAPK) signaling pathway and a key molecular target for ethanol (EtOH) and other drugs of abuse. OBJECTIVE: The aim of the study was to assess the role of two MAPK pathways, ERK(1/2) and c-Jun N terminal kinase (JNK), on the modulation of EtOH and sucrose self-administration. MATERIALS AND METHODS: C57BL/6J mice were trained to lever press on a fixed-ratio 4 schedule with 9% EtOH/2% sucrose, or 2% sucrose, as the reinforcer. In experiments 1 and 2, mice were injected with the MEK(1/2) inhibitor SL 327 (0-100 mg/kg) and the JNK inhibitor AS 6012452 (0-56 mg/kg) prior to self administration. In experiment 3, SL 327 (0-100 mg/kg) was administered prior to performance on a progressive ratio (PR) schedule of EtOH reinforcement. In experiment 4, SL 327 and AS 601245 were injected 2 h before a locomotor test. RESULTS: SL 327 (30 mg/kg) significantly increased EtOH self-administration without affecting locomotion. Higher doses of SL 327 and AS 601245 reduced EtOH reinforced responding and locomotor activity. Reductions of both ligands on sucrose self-administration were due to decreases in motor activity. SL 327 pretreatment had no effect on PR responding. CONCLUSIONS: ERK(1/2) activity is more directly involved in modulating the reinforcing properties of EtOH than JNK activity due to its selective potentiation of EtOH-reinforced responding. The specificity of this effect to EtOH self-administration, rather than sucrose self administration, suggests that the mechanism by which ERK(1/2) increases EtOH reinforced responding does not generalize to all reinforcing solutions and is not due to increased motivation to consume EtOH. PMID- 19125239 TI - Redox cycling in nanofluidic channels using interdigitated electrodes. AB - Amperometric detection is ideally suited for integration into micro- and nanofluidic systems as it directly yields an electrical signal and does not necessitate optical components. However, the range of systems to which it can be applied is constrained by the limited sensitivity and specificity of the method. These limitations can be partially alleviated through the use of redox cycling, in which multiple electrodes are employed to repeatedly reduce and oxidize analyte molecules and thereby amplify the detected signal. We have developed an interdigitated electrode device that is encased in a nanofluidic channel to provide a hundred-fold amplification of the amperometric signal from paracetamol. Due to the nanochannel design, the sensor is resistant to interference from molecules undergoing irreversible redox reactions. We demonstrate this selectivity by detecting paracetamol in the presence of excess ascorbic acid. PMID- 19125238 TI - A therapeutic dose of zolpidem reduces thalamic GABA in healthy volunteers: a proton MRS study at 4 T. AB - BACKGROUND: Zolpidem is a nonbenzodiazepine sedative/hypnotic that acts at GABA(A) receptors to influence inhibitory neurotransmission throughout the central nervous system. A great deal is known about the behavioral effects of this drug in humans and laboratory animals, but little is known about zolpidem's specific effects on neurochemistry in vivo. OBJECTIVES: We evaluated how acute administration of zolpidem affected levels of GABA, glutamate, glutamine, and other brain metabolites. MATERIALS AND METHODS: Proton magnetic resonance spectroscopy ((1)H MRS) at 4 T was employed to measure the effects of zolpidem on brain chemistry in 19 healthy volunteers. Participants underwent scanning following acute oral administration of a therapeutic dose of zolpidem (10 mg) in a within-subject, single-blind, placebo-controlled, single-visit study. In addition to neurochemical measurements from single voxels within the anterior cingulate (ACC) and thalamus, a series of questionnaires were administered periodically throughout the experimental session to assess subjective mood states. RESULTS: Zolpidem reduced GABA levels in the thalamus, but not the ACC. There were no treatment effects with respect to other metabolite levels. Self reported ratings of "dizzy," "nauseous," "confused," and "bad effects" were increased relative to placebo, as were ratings on the sedation/intoxication (PCAG) and psychotomimetic/dysphoria (LSD) scales of the Addiction Research Center Inventory. Moreover, there was a significant correlation between the decrease in GABA and "dizzy." CONCLUSIONS: Zolpidem engendered primarily dysphoric-like effects and the correlation between reduced thalamic GABA and "dizzy" may be a function of zolpidem's interaction with alpha1GABA(A) receptors in the cerebellum, projecting through the vestibular system to the thalamus. PMID- 19125240 TI - CYP3A5 *1 allele associated with tacrolimus trough concentrations but not subclinical acute rejection or chronic allograft nephropathy in Japanese renal transplant recipients. AB - PURPOSE: We assessed reported associations of CYP3A5 *1 allele with a delay in achieving target tacrolimus concentrations, and occurrence of biopsy-confirmed subclinical acute rejection (SAR) and chronic allograft nephropathy (CAN) in Japanese subjects. METHODS: Forty-one renal allograft recipients were studied. The targeted tacrolimus trough concentrations were 20-25 ng/mL up to 2 weeks post transplantation, 10-15 ng/mL up to 6 weeks, and 5-10 ng/mL thereafter. At 1 month and 1 year post-transplantation, allograft biopsies were performed. RESULTS: The CYP3A5 *1/*1 + *1/*3 (expresser) and *3/*3 (nonexpresser) alleles were detected in 19 and 22 patients, respectively. Although the mean trough concentrations were lower in CYP3A5 expressers than nonexpressers for the first 3 weeks, no difference in frequency of SAR among CYP3A5 genotypes was found. The mean trough concentrations were lower from 8 to 12 months post-transplantation, and the frequency of CAN was lower in CYP3A5 expressers. CONCLUSIONS: In contrast to the previous reports, the CYP3A5 *1 allele was not associated with the frequency of SAR or CAN, suggesting that further studies of different immunosuppressive strategies using tacrolimus are needed to confirm the adequate dosing and concentration of tacrolimus for each CYP3A5 genotype. PMID- 19125241 TI - Pharmacokinetics and metabolism of idebenone in healthy male subjects. AB - PURPOSE: Idebenone is a synthetic analogue of ubiquinone that may be beneficial in the treatment of Friedreich's ataxia. Since in previous pharmacokinetic trials only lower doses were studied, it was the aim of this study to evaluate the pharmacokinetics of idebenone in higher doses of up to 2,250 mg/day. METHODS: In this open, randomized trial, 25 healthy male subjects received first either a single oral dose of 150 mg or 750 mg of idebenone, then the same dose given at 8 h intervals for 14 days. RESULTS: Idebenone and its metabolites appeared in the plasma quickly. Over 99% of parent idebenone was metabolized, indicating a high first-pass effect. C(max) and AUC(0-t) values for parent idebenone and its metabolites increased in a dose-proportional manner. There was virtually no accumulation of parent drug or metabolites following multiple dosing. CONCLUSIONS: Idebenone exhibited dose-dependent pharmacokinetics in daily doses up to 2,250 mg. In 6/14 subjects, adverse events of mild to moderate severity were observed. PMID- 19125242 TI - Percutaneous vertebroplasty for multiple myeloma of the cervical spine. AB - INTRODUCTION: Spinal involvement is a common presentation of multiple myeloma (MM); however, the cervical spine is the least common site of myelomatous involvement. Few studies evaluate the results of percutaneous vertebroplasty (PV) in the treatment of MM of the spine. The purpose of this series is to report on the use of PV in the treatment of MM of the cervical spine and to review the literature. MATERIALS AND METHODS: From January 1994 to October 2007, four patients (three men and one woman; mean age, 45 years) who underwent five PV for painful MM in the cervical spine were retrospectively reviewed. The pain was estimated by the patient on a verbal analogic scale. Clinical follow-up was available for all patients (mean, 27.5 months; range, 1-96 months). RESULTS: The mean volume of cement injected per vertebral body was 2.3 +/- 0.8 mL (range, 1.0 4.0 mL) with a mean vertebral filling of 55.0 +/- 12.0% (range, 40.0-75.0%). Analgesic efficacy was achieved in all patients. One patient had a spinal instability due to a progression of spinal deformity noted on follow-up radiographs, without clinical symptoms. Cement leakage was detected in three (60%) of the five treated vertebrae. There was no clinical complication. CONCLUSIONS: The present series suggests that PV for MM of the cervical spine is safe and effective for pain control; nonetheless, the detrimental impact of the disease on bone quality should prompt close radiological follow-up after PV owing to the risk of spinal instability. PMID- 19125243 TI - Automatic determination of Greulich and Pyle bone age in healthy Dutch children. AB - BACKGROUND: Bone age (BA) assessment is a routine procedure in paediatric radiology, for which the Greulich and Pyle (GP) atlas is mostly used. There is rater variability, but the advent of automatic BA determination eliminates this. OBJECTIVE: To validate the BoneXpert method for automatic determination of skeletal maturity of healthy children against manual GP BA ratings. MATERIALS AND METHODS: Two observers determined GP BA with knowledge of the chronological age (CA). A total of 226 boys with a BA of 3-17 years and 179 girls with a BA of 3-15 years were included in the study. BoneXpert's estimate of GP BA was calibrated to agree on average with the manual ratings based on several studies, including the present study. RESULTS: Seven subjects showed a deviation between manual and automatic BA in excess of 1.9 years. They were re-rated blindly by two raters. After correcting these seven ratings, the root mean square error between manual and automatic rating in the 405 subjects was 0.71 years (range 0.66-0.76 years, 95% CI). BoneXpert's GP BA is on average 0.28 and 0.20 years behind the CA for boys and girls, respectively. CONCLUSION: BoneXpert is a robust method for automatic determination of BA. PMID- 19125244 TI - Imaging of accidental paediatric head trauma. AB - Head trauma is the most common form of injury sustained in serious childhood trauma and remains one of the top three causes of death despite improved road planning and safety laws. CT remains the first-line investigation for paediatric head trauma, although MRI may be more sensitive at picking up the full extent of injuries and may be useful for prognosis. Follow-up imaging should be tailored to answer the specific clinical question and to look for possible complications. PMID- 19125245 TI - Gender-specific pattern differences of the ossification centers in the pediatric elbow. AB - BACKGROUND: Only a limited number of studies have investigated the age ranges in which the secondary centers of the elbow appear and ossify. Knowledge of sequence, gender differences and age ranges can aid in accurate assessment of radiographs, especially in cases of injury. OBJECTIVE: To determine the sequence and general age ranges in which each ossification center both appears and fuses, and also to identify differences between genders. MATERIALS AND METHODS: This study included 412 sets of radiographs of children's elbows that were analyzed prospectively by a single experienced pediatric radiologist. The presence as well as state of fusion of each ossification center was noted. The ages of the children ranged from 2 months to 17 years. RESULTS: In girls, the radial head and medial epicondyle appeared at the same age. In boys, there was a trend towards the radial head appearing earlier than the medial epicondyle. There was no statistically significant difference between the age at which the trochlea and olecranon appeared. Our results demonstrate a statistically significant difference between genders in both appearance and fusion. All centers both appeared and fused earlier in girls, with the exception of the appearance of the capitellum. The sequence of appearance and fusion was similar between genders. CONCLUSION: Ossification centers at the elbow both appear and fuse earlier in females but the normal range in age for the times of appearance and fusion of these centers is quite wide for both sexes. PMID- 19125246 TI - Physico-chemical properties of polyhydroxyalkanoate produced by mixed-culture nitrogen-fixing bacteria. AB - Ultra-high molecular weight polyhydroxyalkanoates (PHAs) with low polydispersity index (PDI = 1.3) were produced in a novel, pilot scale application of mixed cultures of nitrogen-fixing bacteria. The number average molecular weight (M (n)) of the poly(3-hydroxybutyrate) (P(3HB)) and poly(3-hydroxybutyrate-co-3 hydroxyvalerate) (P(3HB-co-3HV)) was determined to be 2.4 x 10(6) and 2.5 x 10(6) g mol(-1), respectively. Using two types of carbon sources, biomass contents of the P(3HB) and P(3HB-co-3HV) were 18% and 30% (PHA in dry biomass), respectively. The extracted polymers were analysed for their physical properties using analytical techniques such as nuclear magnetic resonance (NMR) spectroscopy, differential scanning calorimetry (DSC) and gel permeation chromatography (GPC). NMR confirmed the formation of homopolymer and copolymer. DSC showed a single melting endotherm peak for both polymers, with enthalpies that indicated crystallinity indices of 44% and 37% for P(3HB) and P(3HB-co-3HV), respectively. GPC showed a sharp unimodal trace for both polymers, reflecting the homogeneity of the polymer chains. The work described here emphasises the potential of mixed colony nitrogen-fixing bacteria cultures for producing biodegradable polymers which have properties that are very similar to those from their pure-culture counterparts and therefore making a more economically viable route for obtaining biopolyesters. PMID- 19125247 TI - Alcoholic fermentation of xylose and mixed sugars using recombinant Saccharomyces cerevisiae engineered for xylose utilization. AB - Previously, a Saccharomyces cerevisiae strain was engineered for xylose assimilation by the constitutive overexpression of the Orpinomyces xylose isomerase, the S. cerevisiae xylulokinase, and the Pichia stipitis SUT1 sugar transporter genes. The recombinant strain exhibited growth on xylose, under aerobic conditions, with a specific growth rate of 0.025 h(-1), while ethanol production from xylose was achieved anaerobically. In the present study, the developed recombinant yeast was adapted for enhanced growth on xylose by serial transfer in xylose-containing minimal medium under aerobic conditions. After repeated batch cultivations, a strain was isolated which grew with a specific growth rate of 0.133 h(-1). The adapted strain could ferment 20 g l(-1) of xylose to ethanol with a yield of 0.37 g g(-1) and production rate of 0.026 g l(-1) h( 1). Raising the fermentation temperature from 30 degrees C to 35 degrees C resulted in a substantial increase in the ethanol yield (0.43 g g(-1)) and production rate (0.07 g l(-1) h(-1)) as well as a significant reduction in the xylitol yield. By the addition of a sugar complexing agent, such as sodium tetraborate, significant improvement in ethanol production and reduction in xylitol accumulation was achieved. Furthermore, ethanol production from xylose and a mixture of glucose and xylose was also demonstrated in complex medium containing yeast extract, peptone, and borate with a considerably high yield of 0.48 g g(-1). PMID- 19125248 TI - Silencing CD147 inhibits tumor progression and increases chemosensitivity in murine lymphoid neoplasm P388D1 cells. AB - Overexpression of extracellular matrix metalloproteinase inducer (EMMPRIN or CD147), a member glycoprotein enriched on the surface of many malignant tumor cells, promotes tumor progression and confers resistance to some chemotherapeutic drugs. To investigate the possible role of CD147 in the macrophage-like lymphoid neoplasm P388D1 cells progression, we used RNA interference approach to silence CD147 expression. The results showed that silencing of CD147 in P388D1 cells impeded the expression of MMP11 at both mRNA and protein levels. The reduced CD147 expression also resulted in reductions in tumorigenicity, as well as decreased in regional lymph node metastasis. Furthermore, the down-regulation of CD147 expression sensitized cells to be more sensitive to chemotherapeutic drugs. Treatment of tumor cells with U-0126, an inhibitor of mitogen-activated protein kinase/Erk, also down-regulated the expression of MMP11. Our current results indicate that the expression of CD147 functionally mediates tumor progression and is a potential target for therapeutic anti-cancer drugs. PMID- 19125249 TI - A novel beta-thalassemic allele due to a thirteen nucleotide deletion: codons 54 58 (-T ATG GGC AAC CCT). PMID- 19125250 TI - Secondary nodal marginal zone B cell lymphoma arising in primary mixed cellularity subtype of Hodgkin's lymphoma. PMID- 19125251 TI - RAIDD expression is impaired in multidrug resistant osteosarcoma cell lines. AB - PURPOSE: To identify the apoptosis genes involved in the multidrug resistant phenotype of osteosarcoma. METHODS: Multidrug resistant human osteosarcoma cell line (U-2 OS MR) and a drug sensitive parental cell line (U-2 OS) were both treated with paclitaxel and analyzed by the gene array containing 96 apoptosis associated genes. The different expression of the special apoptosis associated genes were further analyzed by Western blot in the multidrug resistant osteosarcoma cell lines (U-2 OS MR, KH OS R2) and the drug sensitive parental cell lines (U-2 OS, KH OS). One of the disregulated gene, RAIDD, was transfected into the multidrug resistant osteosarcoma cells for functional studies. RESULTS: RAIDD showed signs of significant expression in the U-2 OS cells after being treated with paclitaxel (P < 0.01). However, the induction of RAIDD did not occur in U-2 OS MR cells (P = 0.2). Subsequent analysis by Western blot confirmed the deficiency of the expression of RAIDD protein in U-2 OS MR. On the contrary, the expression of RAIDD could be significantly induced by paclitaxel and doxorubicin in U-2 OS cells as both time and dosage were deciding factors. It also demonstrated the cleavage of PARP associated with RAIDD expression in U-2 OS cells, but not however in U-2 OS MR cells after being treated with paclitaxel or doxorubicin. Similar results were found in osteosarcoma multidrug resistant cell line KH OS R2 and the drug sensitive parental cell line KH OS. Furthermore, over expression of RAIDD in multidrug resistant cell lines could possibly reverse drug resistant phenotypes. CONCLUSION: This study indicate that impaired expression of RAIDD in drug induced apoptosis may play a role in the multidrug resistance of osteosarcoma cells. PMID- 19125252 TI - [Work-up of a renal biopsy: what is established? What will follow?]. AB - Percutaneous kidney biopsy now represents the gold standard for diagnosis of most primary and secondary kidney disease. It allows identification and classification of renal disorders and is the basis for standardized therapeutic concepts. As with any diagnostic method, the value of the data produced depends on experience and reproducible procedures. For kidney biopsy this includes resection and adequate methodologies using representative kidney tissue. The procedures required are shown in detail and discussed. PMID- 19125253 TI - Functional analysis of an Arabidopsis transcription factor WRKY25 in heat stress. AB - The WRKY family is one of the major groups of plant-specific transcriptional regulators. Arabidopsis thaliana WRKY25, which is induced by heat stress, is one of the group I WRKY proteins and responds to both abiotic and biotic stress. This study has examined the regulatory role of WRKY25 using wrky25 mutant and over expressing WRKY25 transgenic A. thaliana. After 45 degrees C for different time periods, wrky25 null mutants showed a moderate increase in thermosensitivity with decreased germination, reduced hypocotyl and root growth, and enhanced conductivity compared to those of wide-type, while WRKY25 over-expressed transgenic seeds exhibited enhanced thermotolerance. Northern blot analysis of wrky25 mutants and WRKY25 over-expressing plants identified putative genes regulated by WRKY25. In consistence with the implication of WRKY25 in heat tolerance, the expression level of six heat-inducible genes and two oxidative stress-responsive genes was more or less down-regulated in wrky25 mutants during heat stress. Among them, heat shock protein Hsp101, heat shock transcription factor HsfB2a, and cytosolic ascrobate peroxidase APX1 were reduced more obviously than other detected genes. Meanwhile, over-expression of WRKY25 increased the expression of HsfA2, HsfB1, HsfB2a, and Hsp101 slightly or moderately. Together, these findings reveal that WRKY25 plays a partial role in thermotolerance. PMID- 19125254 TI - Dehydration, rehydration, and overhydration alter patterns of gene expression in the Antarctic midge, Belgica antarctica. AB - We investigated molecular responses elicited by three types of dehydration (fast, slow and cryoprotective), rehydration and overhydration in larvae of the Antarctic midge, Belgica antarctica. The larvae spend most the year encased in ice but during the austral summer are vulnerable to summer storms, osmotic stress from ocean spray and drying conditions due to wind and intense sunlight. Using suppressive subtractive hybridization (SSH), we obtained clones that were potentially responsive to dehydration and then used northern blots to evaluate the gene's responsiveness to different dehydration rates and hydration states. Among the genes most responsive to changes in the hydration state were those encoding heat shock proteins (smHsp, Hsp70, Hsp90), antioxidants (superoxide dismutase, catalase), detoxification (metallothionein, cytochrome p450), genes involved in altering cell membranes (fatty acid desaturase, phospholipase A2 activating protein, fatty acyl CoA desaturase) and the cytoskeleton (actin, muscle-specific actin), and several additional genes including a zinc-finger protein, pacifastin and VATPase. Among the three types of dehydration evaluated, fast dehydration elicited the strongest response (more genes, higher expression), followed by cryoprotective dehydration and slow dehydration. During rehydration most, but not all, genes that were expressed during dehydration continued to be expressed; fatty acid desaturase was the only gene to be uniquely upregulated in response to rehydration. All genes examined, except VATPase, were upregulated in response to overhydration. The midge larvae are thus responding quickly to water loss and gain by expressing genes that encode proteins contributing to maintenance of proper protein function, protection and overall cell homeostasis during times of osmotic flux, a challenge that is particularly acute in this Antarctic environment. PMID- 19125256 TI - Deletion of macrophage migration inhibitory factor attenuates neuronal death and promotes functional recovery after compression-induced spinal cord injury in mice. AB - Macrophage migration inhibitory factor (MIF) is a multipotential protein that acts as a proinflammatory cytokine, a pituitary hormone, and a cell proliferation and migration factor. The objective of this study was to elucidate the role of MIF in spinal cord injury (SCI) using female MIF knockout (KO) mice. Mouse spinal cord compression injury was produced by application of a static load (T8 level, 20 g, 5 min). We analyzed the motor function of the hind limbs and performed histological examinations. Hind-limb function recovered significantly in the KO mice starting from three weeks after injury. Cresyl-violet staining revealed that the number of surviving neurons in the KO mice was significantly larger than that of WT mice six weeks after injury. Immunohistochemical analysis revealed that the number of NeuN/caspase-3-active, double-positive, apoptotic neurons in the KO mice was significantly smaller than that of the WT mice 24 and 72 h after SCI. These results were related to in-vitro studies showing increased resistance of cerebellar granular neurons from MIF-KO animals to glutamate neurotoxicity. These results suggest that MIF existence hinders neuronal survival after SCI. Suppression of MIF may attenuate detrimental secondary molecular responses of the injured spinal cord. PMID- 19125255 TI - Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies. AB - Accumulation of hyperphosphorylated, ubiquitinated and N-terminally truncated TAR DNA-binding protein (TDP-43) is the pathological hallmark lesion in most familial and sporadic forms of FTLD-U and ALS, which can be subsumed as TDP-43 proteinopathies. In order to get more insight into the role of abnormal phosphorylation in the disease process, the identification of specific phosphorylation sites and the generation of phosphorylation-specific antibodies are mandatory. Here, we developed and characterized novel rat monoclonal antibodies (1D3 and 7A9) raised against phosphorylated S409/410 of TDP-43. These antibodies were used to study the presence of S409/410 phosphorylation by immunohistochemistry and biochemical analysis in a large series of 64 FTLD-U cases with or without motor neuron disease including familial cases with mutations in progranulin (n = 5), valosin-containing protein (n = 4) and linkage to chromosome 9p (n = 4), 18 ALS cases as well as other neurodegenerative diseases with concomitant TDP-43 pathology (n = 5). Our data demonstrate that phosphorylation of S409/410 of TDP-43 is a highly consistent feature in pathologic inclusions in the whole spectrum of sporadic and familial forms of TDP 43 proteinopathies. Physiological nuclear TDP-43 was not detectable with these mAbs by immunohistochemistry and by immunoblot analyses. While the accumulation of phosphorylated C-terminal fragments was a robust finding in the cortical brain regions of FTLD-U and ALS, usually being much more abundant than the phosphorylated full-length TDP-43 band, spinal cord samples revealed a predominance of full-length TDP-43 over C-terminal fragments. This argues for a distinct TDP-43 species composition in inclusions in cortical versus spinal cord cells. Overall, these mAbs are powerful tools for the highly specific detection of disease-associated abnormal TDP-43 species and will be extremely useful for the neuropathological routine diagnostics of TDP-43 proteinopathies and for the investigation of emerging cellular and animal models for TDP-43 proteinopathies. PMID- 19125257 TI - Interventions to improve inpatient osteoporosis management following first osteoporotic fracture: the PREVENT project. AB - OBJECTIVES: To establish a protocol for the treatment of fragility fractures in the hospital setting based on treatment of osteoporosis. MATERIALS AND METHODS: An intervention protocol was implemented in patients with fragility fractures based on (1) indicating the diagnosis of osteoporotic fracture in the summary of discharge; (2) "lifestyle recommendations"; and (3) therapy for osteoporosis. Thirty-one hospitals were involved and they were informed of the importance of protocol compliance. In the first phase, a retrospective study was conducted to establish the number of low-energy fractures treated and the percentage of them that had complied with the protocol (n = 887). Then, prospectively, the same data were collected for the patients managed for 1 year (n = 6,826) in three sections of 4-month intervals. RESULTS: The percentage of compliance increased from 8.2 to 57.2% in the first point, from 12.6 to 42.4% in the second, and from 10.3 to 43.2% in the third. CONCLUSION: The implementation of programs to improve osteoporosis treatment is very useful for ensuring adherence in the management of osteoporosis following admission due to fragility fracture. PMID- 19125258 TI - Oncogenic osteomalacia: two case reports with surprisingly different outcomes. AB - Oncogenic osteomalacia is a rare paraneoplastic syndrome of acquired hypophosphatemic osteomalacia, resulting from a deficit in renal tubular phosphate reabsorption, in which fibroblast growth factor 23 (FGF23) seems to be implicated. This condition is usually associated with a phosphaturic mesenchymal tumor of mixed connective tissue located in the bone or soft tissue. The clinical and the radiologic findings are the same as those seen in osteomalacia, and the biochemical features include renal phosphate loss, low serum phosphate and 1,25 (OH)(2) vitD(3) levels, increased alkaline phosphatase, and normal calcium, PTH, calcitonin, 25-OH-vitD(3) and 25,25-(OH)(2) vitD(3). We present two cases of oncogenic osteomalacia associated with phosphaturic mesenchymal tumors, which were histologically similar, but presented a completely different evolution. In the first patient, the tumor developed on the sole of the foot. Following removal of the mass, the symptoms resolved and biochemical and radiological parameters returned to normal. However, in the second patient, a liver tumor developed and resection did not resolve the disease. Multiple lesions appeared in several locations during follow-up. This disease usually remits with complete tumor resection. Nevertheless, if this is not possible, oral treatment with phosphate, calcium and calcitriol can improve the symptoms. If scintigraphy of the tumor shows octreotide receptors, patients may respond partially to therapy with somatostatin analogs, with stabilization of the lesion. PMID- 19125259 TI - Arthroscopically assisted anterior decompression for femoroacetabular impingement: technique and early clinical results. AB - INTRODUCTION: In patients with symptomatic femoroacetabular impingement resection osteochondroplasty of the femoral head-neck junction may improve hip pain and range of motion. We evaluated the short-term treatment results of an arthroscopically assisted mini-open anterior approach to compare it with the results after surgical dislocation for FAI. METHODS: The clinical and radiographic results of 33 patients were reviewed retrospectively 15 months after the surgery. Harris hip scores and plain radiographs were obtained preoperatively and at follow-up. Patient satisfaction with the treatment result was quantified with a Visual Analogous scale (VAS) ranging from 0 (very dissatisfied) to 10 (very satisfied). RESULTS: The mean Harris hip score improved from 64 points preoperatively to 85 points at the time of follow-up (P < 0.001). Mean patient satisfaction on the VAS was seven points (range: 2-10 points). In two of our first patients we observed a transient femoral nerve palsy (completely resolved at follow-up) and 15 patients reported numbness in the area of the lateral cutaneous femoral nerve. CONCLUSIONS: Treatment of anterior femoroacetabular impingement through an arthroscopically assisted mini-open anterior approach can reduce pain and improve function in a short-term observation period. Femoral osteochondroplasty as well as surgical treatment of acetabular cartilage and labrum lesions are possible, but the access is limited to the anterior and anterolateral part of the hip joint. PMID- 19125260 TI - Long-term results of surgical release of trigger finger and trigger thumb in adults. AB - INTRODUCTION: Trigger finger and thumb are amongst the most common hand disabilities seen by orthopaedic surgeons. Surgical release of the first annular pulley (A1) is generally indicated when non-operative treatment fails. We present the long-term results of open operative treatment of trigger finger or trigger thumb in adults performed by a single surgeon. METHOD: From July 1988 to 1998, 305 trigger fingers or thumbs in 276 consecutive patients were treated operatively, including 179 women and 97 men. The average age at operation time was 46.2 years. RESULTS: Two hundred and ten (76%) of the operations were performed for a single trigger digit release, 76 (24%) for multiple trigger digits in individual patients. All operations were done under tourniquet control with local infiltration anaesthesia under outpatient conditions using a transverse incision just distal to the distal palmar crease or on the flexor crease of the thumb at the metacarpophalangeal joint. At latest follow-up after an average of 14.3 years (min: 10, max: 20) 234 patients could be evaluated and were out of complaints, there were no serious complications such as nerve transection or bowstringing or recurrencies. CONCLUSION: We recommend open surgery of trigger finger and trigger thumb. PMID- 19125261 TI - Resurfacing versus nonresurfacing the patella in total knee arthroplasty: a critical appraisal of the available evidence. AB - A systematic literature search for evidences comparing treatment effect and harm of resurfacing versus nonresurfacing the patella in total knee arthroplasty was conducted and yielded five meta-analysis (MA), one systematic review (SR) and six randomized controlled trials not included in previous MAs/SR. The evidence suggests that patellar resurfacing would reduce the risk of anterior knee pain, as well as the risk of patella-related reoperation. Furthermore, patients not undergoing patella resurfacing would experience more knee pain during stair climbing and be less satisfied with surgery. No significant difference in range of motion can be expected with or without patellar resurfacing. Importantly, methodological limitations were observed in all retrieved studies and evidences about potential adverse events related to patellar resurfacing are presently undetermined. PMID- 19125262 TI - Cervical and upper thoracic screwing for spinal fusion: strategy for its safe insertion to avoid major complications. AB - There are several screwing techniques to attain cervical fusion such as pedicle screw, lateral mass screw, facet screw, transarticular and laminar screw. Each screwing technique has advantages and disadvantages. In this study, we introduce our strategy for safe screwing and its clinical results. Our strategy is as follows: lateral mass screw for C1, 3, 4, 5, 6 and pedicle screw for C2, 7, and thoracic level. When the C2 pedicle is thinner than 3.5 mm, we use C2 laminar screws. We do not use Magerl transarticular screw or facet screw; 146 screws were inserted in 17 patients. There were no major complications such as spinal cord and nerve root injury. We did not observe vertebral arterial injury either. Of the 146 screws, 141 (97.0%) were accurately inserted. As for lateral mass screwing by Roy-Camille's technique and C2 laminar screwing, all screws were inserted in the appropriate site (100%) without any complications. Five pedicle screws were misplaced. Of the 57 pedicle screws, 5 showed a minor tear of the wall at C7, Th1 and Th3, the success rate for all pedicle screws was 91%. All showed solid fusion. For cervical screwing the most important aspect should be safety to avoid severe morbidity. Our strategy, which consists in the combined use of pedicle, lateral mass and laminar screwing, is safe and reliable. PMID- 19125263 TI - Prenatal management of an allantoic cyst with patent urachus. AB - Umbilical anomalies are a rare presentation in fetal and neonatal period. We report a case of a patent urachus with an allantoic cyst diagnosed prenatally at 14 weeks of gestation and its spontaneous course throughout ongoing pregnancy. Thoroughly ultrasound examination enabled comprehensive counselling of the parents and together with pediatricians and pediatric surgeons determining the appropriate postnatal surgical management. PMID- 19125264 TI - Effects of two combined oral contraceptives containing ethinyl estradiol 30 microg combined with either gestodene or drospirenone on hemostatic parameters, lipid profiles and blood pressure. AB - OBJECTIVE: The aim of this study is to compare the effect of ethinyl estradiol 0.03 mg/gestodene 0.075 mg (EE/GSD) with ethinylestradiol 0.03 mg/drospirenone 3 mg (EE/DRSP) administered according to conventional 21/7 regimen on body mass index (BMI), blood pressure (BP), lipid metabolism and hemostatic parameters. METHOD: In this study, 160 healthy women were randomized to EE/GSD mg or EE/DRSP for 12 months. Mean differences in BMI, high density lipoprotein-cholesterol (HDL C) and low density lipoprotein-cholesterol (LDL-C), total cholesterol (TC) levels and BP compared to baseline were assessed. RESULTS: One hundred and forty-five (89%) of the women completed all 12 treatment cycles. The subjects randomly assigned into two treatment groups. Group EE/GSD (n = 71) and group EE/DRSP (n = 72). In group B, BMI values were significantly lower than baseline at the sixth cycle. DRSP/EE had more favorable effects on BP than GSD/EE with the mean systolic and diastolic BPs remaining lower in the DRSP/EE group. The difference between the two preparations was not statistically significant at the end of the study. TC levels remained similar in both groups throughout the study period. In both groups LDL-C levels decreased, triglyceride and HDL-C levels significantly increased from baseline levels. These changes result in increasing HDL-C/LDL-C ratio, demonstrating anti-atherogenic effect. Menstrual cycle patterns and the incidence of adverse events were similar between groups. The duration of withdrawal bleeding decreased during the study for both groups and was similar. CONCLUSION: The EE/DRSP regimen provides good cycle control with reliable contraceptive efficacy and low incidence of adverse events. Compared with the EE/GSD preparation, the EE/DRSP preparation demonstrated a more favorable effect on BMI and BP with the mean BMI and mean BP remaining lower than baseline mean. The new formulation may be especially beneficial for women susceptible to body weight gain and rise in BP. PMID- 19125265 TI - Accessory breast tissue presenting as a vulvar mass in an adolescent girl. AB - BACKGROUND: The development of an accessory breast tissue is attributed to the failure of regression of milk line remnants during embryogenesis. The accessory breasts can develop anywhere along the milk line, extending from axilla to the groin. The vulvar accessory breast is extremely rare among these. CASE: We report an 18-year-old girl presenting a solitary vulvar mass which is later pathologically confirmed to be an accessory breast tissue. On preoperative ultrasonographic examination, a lobulated echogenic mass with internal septa was seen in the subcutaneous fat layer of the mons pubis. Excisional biopsy was performed, which revealed an accessory breast tissue. She had no specific developmental abnormalities and had no familial history. CONCLUSION: An accessory breast in the vulva is an extremely rare case among the ectopic breasts, especially in an adolescent girl. Pathological confirmation is mandatory in this case to exclude a possibility of other tumor, including malignancy. PMID- 19125266 TI - The effect of metformin treatment on ovarian stromal blood flow in women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate the effects of metformin on the ovarian stromal blood perfusion in the patients with polycystic ovary syndrome (PCOS). METHODS: Twenty five women with PCOS who underwent a Doppler examination of the ovarian stroma was evaluated; hormonal, anthropometric, and biochemical parameters of patients were determined. After the patients were treated with 850 mg metformin twice a day for 6 months, the same parameters were evaluated in the same patients. RESULTS: After metformin treatment, although pulsatility index (PI) was increased from 1.80 +/- 1.23, 1.84 +/- 1.28 to 2.20 +/- 1.10, 2.19 +/- 0.83 in the right and left ovary, respectively, and resistance index was increased from 0.84 +/- 0.25, 0.83 +/- 0.23 to 1.16 +/- 0.37, 1.10 +/- 0.26 in the right and left ovary respectively (P < 0.05), peak systolic velocity (PSV) was decreased from 12.30 +/ 1.72, 12.34 +/- 1.55 to 10.25 +/- 0.97, 10.53 +/- 1.33 in the right and left ovary respectively (P < 0.05). PI and RI did not show any difference between the homeostatic model assessment insulin resistance index (HOMA-IR) >or= 2.38 and HOMA-IR < 2.38 groups before and after metformin treatment (P > 0.05). However, PSV was decreased significantly from 13.05 +/- 1.35, 12.82 +/- 2.02 to 11.03 +/- 0.71, 10.25 +/- 0.42 in HOMA-IR >or= 2.38 group in the right and left ovary, respectively, and PSV was decreased from 11.50 +/- 2.67, 11.28 +/- 0.26 to 9.10 +/- 0.58, 9.28 +/- 0.25 in HOMA-IR < 2.38 group in the right and left ovary, respectively, before and after metformin treatment (P < 0.05). PSV for both ovaries were positively correlated with HOMA scores before and after metformin treatment [(r = 0.713, P = 0.000; r = 0.617, P = 0.04 and r = 0.635, P= 0.03; r = 0.483, P = 0.031 respectively]. CONCLUSION: Metformin treatment affected ovarian stromal blood flow in PCOS patients. PMID- 19125267 TI - Simultaneous myringoplasty and septoplasty, and the use of nasal septal perichondrium. AB - To investigate the more ideal graft for optimal repair of tympanic membrane perforation, we examined the use of septal perichondrium in myringoplasty. Twenty five patients with ages ranging between 18 and 54 were included in this study. All had a persistent, symptomatic tympanic membrane perforation, and nasal obstruction. All patients were scheduled for myringoplasty by a transmeatal approach in combination with septoplasty under general anaesthesia. Twenty-three patients had intact graft material and had more than 5 dB HL improvement in conductive hearing thresholds at least three frequencies in the follow up period. An overall success rate of 92 and 92% was recorded in terms of hearing improvement and perforation closure, respectively. Nasal septal perichondrium is easily accessible, cost-effective, time saving, sufficiently large, and patient friendly as a graft material in myringoplasty and has a good chance of postoperative survival. PMID- 19125268 TI - Allergic and idiopathic rhinitis: an ultrastructural study. AB - Nasal hyperreactivity is one of the most important underlying mechanisms in both allergic (AR) and idiopathic rhinitis (IR). In order to study the pathomorphological changes in this entity, tissue samples from patients with AR, IR, and from patients without chronic inflammation were taken during nasal surgery. Primary antibodies against Substance P (SP), calcitonin gene-related peptide (CGRP), and endothelial nitric oxide synthases (NOS III) were applied and the immunocomplexes were visualized by immunocytochemistry. The nasal mucosa of patients with AR and IR showed similarities on the ultrastructural level. Neurogenic inflammation was indicated by a strong innervation pattern with sensory nerve fibers containing SP and CGRP. We could show that extensive edema and cellular infiltration might be characteristic for AR. On other hand there was no evidence of eosinophilic or NO involvement in IR. Finally, on the ultrastructural level, AR and IR showed many similarities. Based on these findings anti-inflammatory therapy modalities could be recommended for both types of rhinitis. PMID- 19125269 TI - 3D computer-assisted assessment of complicated penetrating foreign bodies cases in ENT practice. AB - A retrospective research was performed in order to evaluate three-dimensional (3D) computer-assisted detection of penetrating foreign bodies (FB) in the ENT practice in order to assess its usefulness and to specify its application. FBs in the head and neck were detected using 3D CT imaging in order to assess the usefulness of 3D images in the ENT operative practice. Three blinded surgeons were involved in comparison between plain and 3D CT images in order to assess 3D usefulness for precise formulation of a surgical plan. The observed relationship of the FBs to anatomical structures of the ethmoidal sinuses, eye orbit, and neck tissues was found instrumental for surgeon's decision making in planning the approach to operative removal of the FB. It helps to understand the relationships between a FB and surrounding anatomical structures better then the plain X-rays or CT-scan. There was no significant difference in cost between plain CT and 3D images. 3D computer-assisted detection of FBs increase our diagnostic abilities and appears to be a valuable addition to our diagnostic technique. Its main importance, however, lies in its capacity to help a surgeon plan an operation much more carefully avoiding improvisation during the operation itself. PMID- 19125270 TI - Sudden death due to pulmonary embolism from right atrial myxoma. PMID- 19125271 TI - Down-regulation of inducible co-stimulator (ICOS) by intravitreal injection of small interfering RNA (siRNA) plasmid suppresses ongoing experimental autoimmune uveoretinitis in rats. AB - BACKGROUND: RNA interference (RNAi) is now being exploited as a powerful tool for gene knockdown. Recently, we had shown that inducible co-stimulator (ICOS) was up regulated in experimental autoimmune uveoretinitis (EAU). The aim of this study was to investigate whether intravitreal injection of small interfering RNA (siRNA) plasmid, targeting ICOS, suppresses the ongoing experimental autoimmune uveoretinitis (EAU) in rats. METHODS: Oligonucleotide targeting ICOS was cloned into linearized pRNAT-U6.1/Neo eukaryotic expression vector to construct the recombinant plasmid (pRNAT-U6.1/Neo-ICOS). After transfecting activated rat T cells with the recombinant plasmid, ICOS mRNA and protein expression levels were determined by real-time RT-PCR and Western blot analysis respectively. Rats were immunized with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA) and given an intravitreal injection of pRNAT-U6.1/Neo-ICOS on day 6 after immunization. After 13days of immunization, the ICOS protein expression and CD4(+) ICOS (+) T cells were identified in retinae through Western blot analysis and flow cytometry respectively. Intraocular inflammation was assessed by the scores of the clinical and histological appearances. Delayed-type hypersensitivity (DTH) and lymphocyte proliferation were detected to evaluate the systemic effect of intravitreal injection of pRNAT-U6.1/Neo-ICOS. RESULT: The recombinant plasmid (pRNAT-U6.1/Neo-ICOS) for the ICOS siRNA was successfully constructed. In vitro studies using the recombinant plasmid has showed the down regulation of ICOS gene expression both at the mRNA and protein levels. Clinical and pathological scores showed that ocular inflammation of pRNAT-U6.1/Neo-ICOS treated eyes was markedly less than that of vehicle-treated eyes. The expression of ICOS protein and the amount of CD4(+) ICOS(+) T cells in retinae significantly decreased by intravitreal injection of the recombinant plasmid, whereas delayed type hypersensitivity response and lymphocyte proliferation were not impaired in rats treated with the recombinant plasmid. CONCLUSION: Intravitreal injection of siRNA plasmid targeting ICOS effectively down-regulated the expression of ICOS, and was highly effective in suppressing the ongoing process of EAU without any side-effects on systemic cellular immunity. PMID- 19125272 TI - The ascorbic acid transporter SVCT2 is expressed in slow-twitch skeletal muscle fibres. AB - Ascorbic acid, the reduced form of vitamin C, functions as a potent antioxidant as well as in cell differentiation. Ascorbate is taken up by mammalian cells through the specific sodium/ascorbate co-transporters SVCT1 and SVCT2. Although skeletal muscle contains about 50% of the whole-body vitamin C, the expression of SVCT transporters has not been clearly addressed in this tissue. In this work, we analysed the expression pattern of SVCT2 during embryonic myogenesis using the chick as model system. We cloned the chick orthologue of SVCT2 (cSVCT2) that shares 93% identity with the mouse transporter. cSVCT2 mRNA and protein are expressed during chick embryonic muscle development. Immunohistochemical analyses showed that SVCT2 is preferentially expressed by type I slow-twitch muscle fibres throughout chick myogenesis as well as in post-natal skeletal muscles of several species, including human. Our results suggest that SVCT2-mediated uptake of ascorbate is relevant to the oxidative nature of type I muscle fibres. PMID- 19125273 TI - Localization of plasma membrane bound NTPDases in the murine reproductive tract. AB - Extracellular nucleotides might influence aspects of the biology of reproduction in that ATP affects smooth muscle contraction, participates in steroidogenesis and spermatogenesis, and also regulates transepithelial transport, as in oviducts. Activation of cellular nucleotide purinergic receptors is influenced by four plasma membrane-bound members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family, namely NTPDase1, NTPDase2, NTPDase3, and NTPDase8 that differ in their ecto-enzymatic properties. The purpose of this study was to characterize the expression profile of the membrane-bound NTPDases in the murine female and male reproductive tracts by immunological techniques (immunolabelling, Western blotting) and by enzymatic assays, in situ and on tissue homogenates. Other than the expected expression on vascular endothelial and smooth muscle cells, NTPDase1 was also detected in Sertoli cells and interstitial macrophages in testes, in ovarian granulosa cells, and in apical cells from epididymal epithelium. NTPDase2 was largely expressed by cells in the connective tissue; NTPDase3 in secretory epithelia, and finally, NTPDase8 was not detected in any of the tissues studied here. In addition, NTPDase6 was putatively detected in Golgi-phase acrosome vesicles of round spermatids. This descriptive study suggests close regulation of extracellular nucleotide levels in the genital tract by NTPDases that may impact specific biological functions. PMID- 19125274 TI - Self-collected urine sampling to study the kinetics of urinary toluene (and o cresol) and define the best sampling time for biomonitoring. AB - PURPOSE: To study the excretion kinetics of urinary toluene, TOL-U, and o-cresol, o-C, following occupational exposure to toluene in order to define the best time for sample collection, to apply a non-invasive approach based on self-collected urine sampling. METHODS: Five rotogravure printing workers exposed to uncontrolled levels of toluene collected spot urine samples over three consecutive working days and the following day of rest. In each sample TOL-U and o-C were measured and kinetics of excretion evaluated. RESULTS: Toluene exposure ranged from 48.3 to 75.3 mg/m(3); TOL-U and o-C ranged from 1.4 to 34.6 microg/L and from 0.013 to 1.012 mg/L. A time course trend was obtained: TOL-U and o-C increased during the shift and peaked at the end of exposure and up to 2 h later, respectively; afterwards they rapidly decreased following apparent first order kinetics. Considering TOL-U, the elimination half-life for the first fast phase was 79 (+/-35 standard error) min, and for the second slow phase was 1,320 (+/ 1,162) min. For o-C the elimination half-life for the first fast phase was 231 (+/-48) min. Considering a toluene uptake of 86%, TOL-U and o-C excreted in urine were about 0.0067 and 0.18% of the up taken. CONCLUSION: Our results support the use of end shift TOL-U as a short term biomarker of occupational exposure to toluene and show the feasibility of self-collected urine sampling to investigate the elimination kinetics of industrial toxics in humans. PMID- 19125275 TI - Peripheral venous occlusion causing cardiac hypertrophy and changes in biological parameters in rats. AB - Although patients with peripheral vascular disease (PVD) show a high prevalence of coronary artery disease and myocardial hypertrophy, influences of PVD alone on cardiac hypertrophy, and its subcellular mechanism remain unclear. This study investigates the influences of PVD alone on cardiac muscles and the changes in biological parameters that potentially affect cardiac hypertrophy and cardioprotection. Several veins from rat hind limb were occluded. Two weeks later, the weight of heart relative to body weight in the occluded group increased by approximately 10% compared to that in the sham-operated group. A significant increase was observed in insulin-like growth factor-1 expression, and a significant decrease was observed in myostatin and mechano growth factor in the occluded group compared to the sham-operated group. Tissue concentration of nitric oxide, and content of heat shock protein (HSP)-72, HSP-90, and heat shock cognate protein-70 significantly decreased in the hypertrophied myocardium. These results suggest that peripheral occlusion alone causes cardiac hypertrophy associated with changes in biological parameters. PMID- 19125276 TI - Gender influences neuromuscular adaptations to muscle unloading. AB - Muscle unloading results in decreased strength. This is partly attributed to reduced neural activation. This study examined whether men and women experienced different adaptations to muscle unloading. Ten men (21.4 +/- 0.8 years; mean +/- SE) and ten women (20.9 +/- 0.2 years) were subjected to 1 week of muscle unloading. Muscle function was quantified during knee extensions before, and following unloading. Electromyography (EMG) was assessed during maximal isometric contractions before, and after unloading. Results showed greater (P < 0.05) unloading-induced decrement in muscle performance in women than men. The loss of peak torque was significantly correlated (r = 0.69) with the decline in EMG consequent to unloading, and women displayed significantly larger reductions in EMG. These findings indicate that: (1) women are more affected by muscle unloading than men, and (2) gender-related differences in strength declines are associated with similar disparities in the nervous system's capacity to maximally stimulate muscle. PMID- 19125277 TI - Association of angiotensin-converting enzyme insertion/deletion polymorphism with obesity, cardiovascular risk factors and exercise-mediated changes in Korean women. AB - This study examined whether the angiotensin-converting enzyme (ACE) insertion (I)/deletion (D) polymorphism is associated with obesity, cardiovascular risk factors and 12-week exercise-mediated changes in Korean women. A total of 105 subjects were divided into three groups as II, ID and DD genotype groups based upon ACE I/D genotypes. Body composition and cardiovascular risk factors were compared among the three groups, and the association of ACE I/D genotypes with obesity and hypertension was evaluated. Total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) levels were higher (P < 0.05) in the DD genotype than in II or ID genotypes. D allele frequency in ACE I/D gene had a higher (P = 0.063) trend in the hypertensive group than the normotensive group. The DD genotype had a trend to develop (odds ratio 4.032, P = 0.086) more hypertension than the II genotype. The II and ID genotypes showed a significant (P < 0.05) decrease in intima media thickness of the carotid artery after an exercise intervention, whereas the DD genotype showed an increase. In conclusion, there is a trend towards association of ACE I/D polymorphism with hypertension but not with obesity. Exercise-mediated changes did not differ significantly among genotypes except IMTCA. PMID- 19125278 TI - Comments on the paper "End-tidal pressure of CO2 and exercise performance in healthy subjects" by Bussotti M, Magri D, Previtali E, Farina S, Torri A, Matturri M, Agostoni P (2008) Eur J Appl Physiol 103(6):727-732. PMID- 19125279 TI - A comparison of different two-dimensional approaches for the determination of the patellar tendon moment arm length. AB - The purpose of this study was to estimate and compare the moment arm length of the patellar tendon (d) during passive knee extension using three different reference landmarks; instant centre of rotation (ICR), tibiofemoral contact point (TFCP) and geometrical centre of the posterior femoral condyles (GCFC). Measurements were taken on the right leg on seven healthy males during passive knee rotation performed by the motor of a Cybex Norm isokinetic dynamometer. Moment arms lengths were obtained by analysing lateral X-ray images recorded using a GE FlexiView 8800 C-arm videofluoroscopy system. The d-knee joint angle relations with respect to GCFC and ICR were similar, with decreasing values from full knee extension (~5.8 cm for d (GCFC) and ~5.9 cm for d (ICR)) to 90 degrees of knee flexion (~4.8 cm for both d (GCFC) and d (ICR)). However, the d (TFCP) knee joint angle relation had an ascending-descending shape, with the highest d (TFCP) value (~5 cm) at 60 degrees of knee flexion. There was no significant difference between the GCFC and ICR methods at any knee joint angle. In contrast, there were significant differences (P < 0.01) between d (ICR) and d (TFCP) at 0 degrees , 15 degrees , 30 degrees and 45 degrees of knee flexion and between d (GCFC) and d (TFCP) at 0 degrees , 15 degrees and 30 degrees of knee flexion (P < 0.01). This study shows that when using different knee joint rotation centre definitions, there are significant differences in the estimates of the patellar tendon moment arm length, especially in more extended knee joint positions. These differences can have serious implications for joint modelling and loading applications. PMID- 19125280 TI - Exercise training reduces cardiac angiotensin II levels and prevents cardiac dysfunction in a genetic model of sympathetic hyperactivity-induced heart failure in mice. AB - The role of exercise training (ET) on cardiac renin-angiotensin system (RAS) was investigated in 3-5 month-old mice lacking alpha(2A-) and alpha(2C-)adrenoceptors (alpha(2A)/alpha(2C)ARKO) that present heart failure (HF) and wild type control (WT). ET consisted of 8-week running sessions of 60 min, 5 days/week. In addition, exercise tolerance, cardiac structural and function analysis were made. At 3 months, fractional shortening and exercise tolerance were similar between groups. At 5 months, alpha(2A)/alpha(2C)ARKO mice displayed ventricular dysfunction and fibrosis associated with increased cardiac angiotensin (Ang) II levels (2.9-fold) and increased local angiotensin-converting enzyme activity (ACE 18%). ET decreased alpha(2A)/alpha(2C)ARKO cardiac Ang II levels and ACE activity to age-matched untrained WT mice levels while increased ACE2 expression and prevented exercise intolerance and ventricular dysfunction with little impact on cardiac remodeling. Altogether, these data provide evidence that reduced cardiac RAS explains, at least in part, the beneficial effects of ET on cardiac function in a genetic model of HF. PMID- 19125281 TI - A simple theoretical model of heat and moisture transport in multi-layer garments in cool ambient air. AB - Overall resistances for heat and vapor transport in a multilayer garment depend on the properties of individual layers and the thickness of any air space between layers. Under uncomplicated, steady-state conditions, thermal and mass fluxes are uniform within the garment, and the rate of transport is simply computed as the overall temperature or water concentration difference divided by the appropriate resistance. However, that simple computation is not valid under cool ambient conditions when the vapor permeability of the garment is low, and condensation occurs within the garment. Several recent studies have measured heat and vapor transport when condensation occurs within the garment (Richards et al. in Report on Project ThermProject, Contract No. G6RD-CT-2002-00846, 2002; Havenith et al. in J Appl Physiol 104:142-149, 2008). In addition to measuring cooling rates for ensembles when the skin was either wet or dry, both studies employed a flat-plate apparatus to measure resistances of individual layers. Those data provide information required to define the properties of an ensemble in terms of its individual layers. We have extended the work of previous investigators by developing a rather simple technique for analyzing heat and water vapor transport when condensation occurs within a garment. Computed results agree well with experimental results reported by Richards et al. (Report on Project ThermProject, Contract No. G6RD-CT-2002-00846, 2002) and Havenith et al. (J Appl Physiol 104:142-149, 2008). We discuss application of the method to human subjects for whom the rate of sweat secretion, instead of the partial pressure of water on the skin, is specified. Analysis of a more complicated five-layer system studied by Yoo and Kim (Text Res J 78:189-197, 2008) required an iterative computation based on principles defined in this paper. PMID- 19125282 TI - Effect of the leg muscle pump on the rise in muscle perfusion during muscle work in humans. AB - The transient and steady-state effects of the calf muscle pump on the rise in muscle perfusion during rhythmic plantarflexions were investigated in 20 volunteers. Because a large hydrostatic column would increase the effect of a muscle pump, exercise in the supine and head-up tilted positions was compared. Within approximately 15 s of the start of muscle work, femoral artery flow (ultrasound Doppler) rose 0.37 L/min above rest in the supine and 0.5 L/min above rest in the tilted position. The latter is a significantly larger rise (P < 0.05). After 80 s of muscle work, femoral flow was stable at 0.38 and 0.39 L/min above rest in the supine and tilted positions, respectively. We conclude that the muscle pump contributes to muscle perfusion during the initial phase of muscle work, but that metabolic vasodilation is a more important determinant of muscle perfusion during steady-state muscle work. PMID- 19125283 TI - Effect of acute sprint interval exercise on central and peripheral artery distensibility in young healthy males. AB - Peripheral arterial distensibility is improved with sprint interval exercise training in young healthy participants (Rakobowchuk et al. in Am J Physiol Regul Integr Comp Physiol 295:R236-R242, 2008). To fully understand the mechanisms contributing to these training effects it is useful to examine the acute responses to sprint interval exercise. Following supine rest, nine healthy males completed either a single sprint interval (Wingate test) or a multiple sprint interval exercise session (4 Wingate tests each separated by 4.5 min). Following exercise, participants recovered for 60 min while central and peripheral arterial distensibility measurements were conducted at discrete time points, using applanation tonometry and ultrasound imaging and continuously, using central and peripheral pulsewave velocity (PWV). Single and multiple sprint interval exercise sessions caused similar changes in all variables. Heart rate was increased throughout recovery (p < 0.05), while central artery PWV was increased until 20 min of recovery (p < 0.05) and lower extremity PWV was decreased until ~45 min (p < 0.05). Distensibility of the superficial femoral artery showed a trend for a reduction at 2 min post-exercise (p = 0.06). These results indicate that extremely high intensity exercise transiently increases central artery stiffness, while metabolite induced vasodilation reduces peripheral stiffness in exercised limbs well into recovery. PMID- 19125284 TI - Total haemoglobin mass but not cardiac volume adapts to long-term endurance exercise in highly trained spinal cord injured athletes. AB - The oxygen transport system is an important component in the limitation of endurance performance in able-bodied and paraplegic athletes. The aim of the present study was to investigate the total haemoglobin mass (tHb, carbon monoxide rebreathing method) and cardiac volume (HV, echocardiography) in 25 highly endurance trained male spinal cord injured (mainly paraplegic) athletes (SCI TRAINED) and to compare the results with those of 10 untrained spinal cord injured controls (SCI-UNTRAINED) and in 25 able-bodied elite endurance athletes (TRAINED). tHb and tHb/kg were higher in SCI-TRAINED than in SCI-UNTRAINED (748 +/- 110 vs. 629 +/- 209 g (464 +/- 68 vs. 390 +/- 130 mmol) (mean +/- SD), P = 0.02 and 10.3 +/- 1.3 vs. 7.9 +/- 2.0 g/kg (6.4 +/- 0.8 vs. 4.9 +/- 1.2 mmol/kg), P < 0.0001), while HV and HV/kg showed no significant differences between the two groups (765 +/- 93 vs. 793 +/- 164 ml and 10.6 +/- 1.4 vs. 10.3 +/- 2.5 ml/kg). No difference between SCI-TRAINED and TRAINED was found for septal diameter (9.5 +/- 1.0 mm vs. 9.7 +/- 0.7 mm). However, tHb and tHb/kg in SCI-TRAINED was lower than in TRAINED [896 +/- 123 g (556 +/- 76 mmol), P = 0.0003 and 12.6 +/- 1.3 g/kg (7.8 +/- 0.8 mmol), P < 0.0001]. In spinal cord injured athletes, tHb but not HV adapts moderately to chronic endurance exercise, although tHb in spinal cord injured athletes does not reach the level of able-bodied-trained persons. PMID- 19125285 TI - Particulate matter inhalation during hay storing activity induces systemic inflammation and platelet aggregation. AB - The aim of this study was to investigate possible pathomechanisms behind the cardiovascular morbidity caused by inhalation of particulate matter (PM(10)). For that purpose, healthy volunteers were exposed to high PM(10) concentrations during a 2 h hay storing activity. Blood was drawn in the evening before and after PM(10) exposure and in the morning and evening of the day after exposure. The leukocyte count increased after PM(10) exposure with an initial increase of segmented neutrophils followed by banded forms. C-reactive protein increased over time. Fibrinogen and plasma viscosity became increased in the evening of the day after PM(10) exposure. Platelet aggregation was increased in the evening after PM(10) exposure. At the same time von Willebrand factor and factor VIII were increased, reflecting endothelial activation. These results confirm that acute inhalative exposure to high PM(10) concentrations during hay storage activity leads to a systemic inflammatory reaction, endothelial activation, and platelet aggregation. PMID- 19125286 TI - Changes in serum cartilage oligomeric matrix protein (COMP), plasma CPK and plasma hs-CRP in relation to running distance in a marathon (42.195 km) and an ultra-marathon (200 km) race. AB - Marathon running is frequently associated with numerous cellular changes, but little information is available on the effects of exercise-mediated prolonged impact-stress on cartilage integrity. This study was undertaken to evaluate muscle and cartilage damage with different running distances. Twenty male marathoners and ultra-marathoners participated in the study. Serum COMP and plasma CPK and hs-CRP were measured as markers of cartilage and muscle damage and inflammation. Serum COMP was increased 1.6-fold at 10 km during a marathon race and declined to the pre-race level after 2 days recovery. In contrast, serum COMP was increased 1.9-fold after a 200-km race and maintained until day 3 of recovery, only returning to the pre-race level on day 6. Plasma CPK was increased at 10 km of the marathon race and up to threefold at the end of the race. This was further increased on day 1, only returning to pre-race level on day 6. Plasma CPK was increased 35-fold at the end of the 200-km race and remained increased until day 5. There was no change in plasma hs-CRP during the marathon race, but this was increased 3.4-fold by day 1, returning to the pre-race level on day 4. Plasma hs-CRP increased 40-fold by the end of the 200-km race and was still increased on day 6 of recovery. Therefore, longer distance running may induce more impact-stress both on muscle and cartilage. Further, the required time for recovery may vary with running distance and the tissue type, e.g. cartilage or skeletal muscle as in this case. PMID- 19125287 TI - Effect of two durations of short-term intermittent hypoxia on ventilatory chemosensitivity in humans. AB - The purpose of this study was to clarify the influence of duration of intermittent hypoxia per day on ventilatory chemosensitivity. Subjects were assigned to three different groups according to the duration of exposure to intermittent hypoxia (12.3 +/- 0.2% O(2)): a first group (H-1, n = 6) was exposed to hypoxia for 1 h per day, the second group (H-2, n = 6) was exposed for 3 h per day, and the third (C, n = 7) was used as control. Hypoxic and hypercapnic ventilatory responses (HVR and HCVR) were determined before and after 1 week of intermittent hypoxia. HVR was increased significantly (P < 0.05) after intermittent hypoxia in both the H-1 and H-2 groups. However, there was no significant difference in magnitude of increased HVR between H-1 and H-2 groups. HCVR did not show any changes in all groups after intermittent hypoxia. These results suggest that 1 h of daily exposure is as equally effective as 3 h of daily exposure to severe hypoxia for a short period for enhancing ventilatory chemosensitivity to hypoxia. PMID- 19125288 TI - PtFLC homolog from trifoliate orange (Poncirus trifoliata) is regulated by alternative splicing and experiences seasonal fluctuation in expression level. AB - In many plant species, exposure to a prolonged period of low temperature during the winter promotes flowering in the spring, a process termed vernalization. In Arabidopsis, the vernalization requirement of winter annual ecotypes is caused by a MADS-box gene FLOWERING LOCUS C (FLC), which is a repressor of flowering gene. Here, a MADS-box gene was isolated from an early flowering trifoliate orange mutant (precocious trifoliate orange, Poncirus trifoliata L. Raf) by the RACE method combined with a cDNA library. Phylogenetic analysis reveals that the MADS box gene is more closely related to the homologs of the FLOWERING LOCUS C lineage than to any of the other MIKC-type MADS-box lineages known from Arabidopsis. The expression profile of the MADS-box gene by real-time PCR showed upregulation of PtFLC during the winter, followed by a decrease in the spring and summer. This kind of cycling is contrary to the pattern observed in Arabidopsis. In situ hybridization reveals that the MADS-box gene is predominately expressed in the vegetative and reproductive meristems. In addition, five alternatively spliced transcripts of the MADS-box gene were also isolated at juvenile and adult mutant developmental stages. Expression analysis of these transcripts at different developmental stages indicated involvement of alternative splicing during phase change. The information suggests a complicated regulation mechanism in seasonal response and flower formation in perennial woody plants. PMID- 19125289 TI - Heat-inducible C3HC4 type RING zinc finger protein gene from Capsicum annuum enhances growth of transgenic tobacco. AB - Capsicum annuum RING Zinc Finger Protein 1 (CaRZFP1) gene is a novel C3HC4-type RING zinc finger protein gene which was previously isolated from a cDNA library for hot pepper plants treated of heat-shock. The CaRZFP1 was inducible to diverse environmental stresses in hot pepper plants. We introduced the CaRZFP1 into the Wisconsin 38 cultivar of tobacco (Nicotiana tabacum) by Agrobacterium mediated transformation under the control of the CaMV 35S promoter. Expression of the transgene in the transformed tobacco plants was demonstrated by RNA blot analyses. There appeared no adverse effect of over-expression of the transgene on overall growth and development of transformants. The genetic analysis of tested T(1) lines showed that the transgene segregated in a Mendelian fashion. Transgenic tobacco lines that expressed the CaRZFP1 gene were compared with several different empty vector lines and they exhibited enhanced growth; they have larger primary root, more lateral root, larger hypocotyls and bigger leaf size, resulting in heavier fresh weight. Enhanced growth of transgenic lines accompanied with longer vegetative growth that resulted in bigger plants with higher number of leaves. Microarray analysis revealed the up-regulation of some growth related genes in the transgenic plants which were verified by specific oligomer RNA blot analyses. These results indicate that CaRZFP1 activates and up regulates some growth related proteins and thereby effectively promoting plant growth. PMID- 19125290 TI - Myxoid solitary fibrous tumor: a clinicopathologic study of three cases. AB - While focal myxoid areas are occasionally observed in solitary fibrous tumors, neoplasms of this type exhibiting extensive myxoid change are considered exceedingly uncommon. Due to their rarity, the biologic behavior of myxoid solitary fibrous tumor has not been determined. Three cases of myxoid solitary fibrous tumor are described in order to better characterize the clinical and pathologic features of this uncommon variant of solitary fibrous tumor. The tumors occurred in one man and two women, with ages of 37, 47, and 58 years, respectively. Sites of involvement included the retroperitoneum, pelvis, and soft tissue of the neck. Histologically, all cases were characterized predominantly by the presence of myxoid stroma comprising 70% to 100% of the tumor. The tumor cells were predominantly spindled in all cases, and arranged randomly, in loose fascicles, or in anastomosing strands imparting a microcystic/reticular appearance. The lesional cells had a bland cytologic appearance and low mitotic count. All tumors lacked necrosis and areas of increased cellularity. By immunohistochemistry, all cases were positive for CD34, CD99, and bcl-2, and negative for keratin, epithelial membrane antigen, desmin, actin, smooth muscle actin, and S-100 protein. To date, all cases have followed a benign course without evidence of recurrence or metastasis with a follow-up duration ranging from 50 to 87 months. The data suggest that myxoid solitary fibrous tumors are associated with an indolent clinical course and favorable prognosis. PMID- 19125291 TI - Histoplasma-associated inflammatory pseudotumour of the kidney mimicking renal carcinoma. AB - A 56-year-old female, originally from Suriname, with an otherwise unremarkable previous medical history was found to have a renal mass highly suspicious for renal cancer for which a nephrectomy was performed. Within the kidney, a tumourous mass was found which, on histological examination, showed an inflammatory pseudotumour caused by Histoplasma capsulatum. Further investigations revealed an idiopathic CD4(+) lymphopenia. Mass lesions mimicking a malignant tumour caused by infection with Histoplasma have rarely been described. To the best of our knowledge, this is the first report of a Histoplasma-associated inflammatory pseudotumour mimicking cancer occurring in the kidney. PMID- 19125292 TI - Improved general health of international adoptees, but immunization status still insufficient. AB - We studied the demographic and clinical data from 495 adopted children seen between January 2002 and January 2007 to evaluate the medical condition and immunization status of international adoptees. The data of children from Chinese origin (53.5%) were compared to children arriving from other countries. Medical problems requiring treatment were present in 42.8% of the children. Parasitic gastrointestinal infection (22.0%) and skin abnormalities (22.4%) were diagnosed most often. Hepatitis B (1.2%) and tuberculosis (1%) were documented in some children; HIV, hepatitis C, and syphilis were not seen in any of the children. Antibody levels against diphtheria and tetanus were insufficient in about half of all children, particularly in those from China. In conclusion, most adoptive children had a good general health, with only a few having major medical problems. Many adoptive children had an inadequate immunization status. PMID- 19125293 TI - Plagiarism and other scientific misconducts. PMID- 19125294 TI - Down-regulation of ribosomal protein L7A in human osteosarcoma. AB - PURPOSE: To explore the expression of ribosomal protein L7A (RPL7A) in osteosarcoma and its correlation with clinical features. METHODS: Ribosomal protein L7A mRNA expression was quantified by real-time reverse transcription (RT)-polymerase chain reaction (PCR) in 47 specimens from osteosarcoma, 8 from normal bone tissues and 12 from benign bone lesion tissues. Expression of RPL7A mRNA was also detected in human osteosarcoma cell line MG-63. The relationship between RPL7A mRNA expression and clinicopathological factors was statistically analyzed. The immunoblotting pattern of RPL7A was also analyzed in 20 osteosarcomas, 8 normal bone and 8 benign bone tissues. RESULTS: Ribosomal protein L7A mRNA expression in osteosarcoma samples was significantly down regulated compared with that in samples from normal bone (P < 0.001) and benign bone lesion tissues (P < 0.001). Low expression of RPL7A mRNA was also found in osteosarcoma cell line MG-63. Low expression of RPL7A mRNA was significantly associated with increased serum level of alkaline phosphatase (ALP, P = 0.008), but was not correlated with other clinicopathological parameters including sex, age, serum lactate dehydrogenase (LDH), tumor location, histological type, histological grading, lung metastasis and overall survival. Interestingly, survival analysis suggested low RPL7A mRNA expression was a significant poor prognostic indicator for overall survival in patients with high grade lesion developed lung metastasis at the time of diagnosis of the primary osteosarcoma (P < 0.05). On western blot, reduced expression of RPL7A protein was observed in osteosarcoma samples (n = 20) compared with normal bone (n = 8) (P < 0.001) and benign bone tissues (n = 8) (P < 0.001). CONCLUSION: Under-expression of RPL7A may be involved in the carcinogenesis of osteosarcoma. Loss of RPL7A expression may be associated with poor survival of osteosarcoma patients with lung metastasis. PMID- 19125295 TI - Progressive tumor growth-associated altered tumor microenvironment: implications in a tumor stage-dependent modulation in survival of a murine T cell lymphoma. AB - In the present study using a transplantable murine T cell lymphoma designated as Dalton's lymphoma, we investigated the role of oxygen, glucose and other environmental factors in evolution of altered survival responses in tumor cells during the late tumor-bearing stages. Tumor progression was observed to be associated with an improvement in the survival ability of tumor cells. Moreover, tumor serum and ascitic fluid obtained from the late tumor bearing stage was found to augment tumor cell survival in vitro, indicating that these humoral components of tumor bearing host contain factors capable of modulating tumor survival. Progressive tumor growth was also shown to be associated with depletion in glucose and oxygen content in the fluids of tumor microenvironment along with a concomitant augmentation in the production of lactate and lactate dehydrogenase. Moreover, tumor cells expressed higher amount of mRNA for inducible nitric oxide synthase, Hypoxia inducible factor-1 and 2 and Hsp70 and VEGF proteins during the late tumor bearing stages. Splenic macrophages and non adherent splenic lymphocytes from tumor-bearing mice showed an increased production of IL-6, TGF-beta, IFN-gamma, IL-2R and VEGF during the late tumor bearing stage, which could be implicated in the differential regulation of tumor growth in a tumor stage dependent manner. In conclusion, the observations of the present study suggest that factors contained in altered tumor microenvironment may act in concert to cause behavioral alterations in tumor cells, with respect to survival, during the course of the progression of a nonsolid T cell lymphoma. PMID- 19125296 TI - Gene expression of ceramide kinase, galactosyl ceramide synthase and ganglioside GD3 synthase is associated with prognosis in breast cancer. AB - PURPOSE: Sphingolipids are bioactive lipids implicated in apoptosis, cell survival and proliferation. We analyzed the prognostic value of enzymes from sphingolipid metabolism in breast cancer. METHODS: Differences in expression of ceramide galactosyl transferase (UGT8), ceramide kinase (CERK), and Ganglioside GD3-Synthase (ST8SIA1) in breast cancer cells were investigated by using microarray data of 1,581 tumor samples. RESULTS: UGT8, CERK, and ST8SIA1 were associated with poor pathohistological grading (P < 0.001). High CERK expression was correlated with ErbB2 status (P = 0.006). Among ER positive breast cancers a significant worse prognosis for patients with tumors showing low ST8SIA1 and UGT8 expression was observed. In the ER negative subgroup those samples with high CERK expression displayed a worse prognosis. In a multivariate analysis only ST8SIA1 and tumor size remained significant. CONCLUSIONS: Our experiments reveal that expression of enzymes from the sphingolipid metabolism has prognostic implications in breast cancer. PMID- 19125297 TI - Plasma malondialdehyde levels and CXCR4 expression in peripheral blood cells of breast cancer patients. AB - INTRODUCTION: Breast cancer is one of the most common neoplasms in women and is a leading cause of cancer related deaths worldwide. Chemokines and their receptors are involved in the control of lymphocyte traffic, a critical component of systemic immunity. CXCR4 mRNA could be involved in the development of variety of diseases. Lipid peroxidation, the result of nonenzymatic autooxidation of polyunsaturated fatty acids, presents numerous harmful effects on biological systems and has been implicated in diseases like cancer. This study examined CXCR4 mRNA expression in peripheral blood cells and malondialdehyde (MDA) in plasma from blood donors and breast cancer patients. MATERIALS AND METHODS: CXCR4 expression in peripheral blood cells from 59 breast cancer patients and 76 healthy blood donors was analyzed by real-time PCR. Plasma MDA was analyzed using high-performance liquid chromatography (HPLC). CONCLUSION: In all stages, MDA levels in total breast cancer patients (1.41 +/- 0.11) were significantly higher (P < 0.01) than those in healthy subjects (0.34 +/- 0.03). No statistically significant difference occurred between CXCR4 expression in peripheral blood cells from breast cancer patients (1.69 +/- 1.05) and the normal healthy control group (1.8 +/- 0.65). However, stage II samples differed statistically (4.3 +/- 1.72) from control, total cancer patients and stages I, III and IV samples. PMID- 19125298 TI - Methylation status of p16 INK4A tumor suppressor gene in Iranian patients with sporadic breast cancer. AB - INTRODUCTION: p16(INK4A) is a tumor suppressor encoding the Cdk inhibitor protein, which acts to repress Cdk4/6 and pRb phosphorylation. p16(INK4A) gene can be inactivated by a variety of events, including promoter hypermethylation. MATERIALS AND METHODS: To investigate the methylation status of the p16(INK4A) gene in Iranian patients with breast carcinoma, promoter methylation was studied by methylation-specific PCR (MSP) and restriction enzyme-related PCR (REP). In addition, p16(INK4A) promoter was analyzed by PCR-SSCP in order to detection of mutation and single nucleotide polymorphisms. RESULTS: Analysis of 70 patients by MPS and REP showed hypermethylation of p16(INK4A) promoter in 35.7% (25/70) and 40% (28/70) of samples, respectively. Comparison of the molecular data and pathological information of the samples suggested that p16(INK4A) gene might be inactivated at the early stages in breast cancer. CONCLUSION: Therefore, it could be suggested that hypermethylation of p16(INK4A) promoter is one of the epigenetic factors affecting the progress of sporadic breast carcinogenesis in Iranian patients. PMID- 19125299 TI - Mitochondrial DNA content in paired normal and cancerous breast tissue samples from patients with breast cancer. AB - INTRODUCTION: We develop a multiplex quantitative real-time PCR for synchronized analysis of mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) to investigate relative mtDNA abundance in paired normal and cancerous breast tissues. MATERIALS AND METHODS: The amounts of nDNA and mtDNA in 102 tissue samples were quantified for both glyceraldehype-3-phosphodehydrogenase (GAPDH) gene and mtDNA encoded ATPase (MTATP) 8 gene. The average threshold cycle (Ct) number values of the nDNA and mtDNA were used to calculate relative mtDNA content in breast tissues. RESULTS: The median delta Ct (DeltaCt) and the median mtDNA content for normal and cancerous breast tissues were 6.73 and 2.54, as well as 106.50 and 5.80 (P = 0.000, respectively). The mtDNA content was decreased in 82% of cancerous breast tissues compared with the normal ones. The changes were associated with hormone receptor status. CONCLUSION: Our finding suggests that decreased mtDNA content in breast cancer may have diagnostic and prognostic value for the disease. PMID- 19125300 TI - Clinical implications of molecular genetic aberrations in acute myeloid leukemia. AB - The role of different cytogenetic changes has been extensively evaluated in patients with acute myeloid leukemia (AML), and cytogenetic analysis of AML blasts is essential to form prognostic subgroups in order to stratify for the extent of therapy. Nevertheless, 40-45% of AML patients lack such cytogenetic markers, i.e., cytogenetically normal AML (CN-AML). In the past decade, different molecular aberrations were identified in AML and especially CN-AML can now be discriminated into certain prognostic subgroups. This review considers the latest advances to define the prognostic impact of molecular aberrations in AML and gives insights how such molecular markers can be applied for analysis of minimal residual disease. Furthermore, therapeutic implications as well as the potential role of new methodological techniques in analyzing expression patterns of AML blasts are discussed. PMID- 19125301 TI - Analysis of adaptation to high ethanol concentration in Saccharomyces cerevisiae using DNA microarray. AB - In industrial process, yeast cells are exposed to ethanol stress that affects the cell growth and the productivity. Thus, investigating the intracellular state of yeast cells under high ethanol concentration is important. In this study, using DNA microarray analysis, we performed comprehensive expression profiling of two strains of Saccharomyces cerevisiae, i.e., the ethanol-adapted strain that shows active growth under the ethanol stress condition and its parental strain used as the control. By comparing the expression profiles of these two strains under the ethanol stress condition, we found that the genes related to ribosomal proteins were highly up-regulated in the ethanol-adapted strain. Further, genes related to ATP synthesis in mitochondria were suggested to be important for growth under ethanol stress. We expect that the results will provide a better understanding of ethanol tolerance of yeast. PMID- 19125302 TI - Study of kinetic parameters in a mechanistic model for bioethanol production through a screening technique and optimization. AB - The accurate description of the kinetics and robust modeling of biotechnological processes can only be achieved by incorporating reliable methodologies to easily update the model when there are changes in operational conditions. The purpose of this work is to provide a systematic approach with which to perform model parameters screening and updating in biotechnological processes. Batch experiments are performed to develop a mechanistic model, considering the effect of temperature on the kinetics, and further experiments (batch fermentations using sugar cane molasses from a different harvest) are used to validate the effectiveness of screening before parameters updating. The reduction in the number of kinetic parameters to be re-estimated enabled by the screening procedure reduces significantly the complexity of the optimization, which makes the updating procedure to be significantly quicker, while resulting in accurate performance of the updated model. PMID- 19125303 TI - Expression of the nitrate transporter nrt2 gene from the symbiotic basidiomycete Hebeloma cylindrosporum is affected by host plant and carbon sources. AB - Although the function of the extramatrical mycelium of ectomycorrhizal fungi is considered essential for the acquisition of nitrogen by forest trees, gene regulation in this fungal compartment is poorly characterized. In this study, the expression of the nitrate transporter gene nrt2 from the ectomycorrhizal basidiomycete Hebeloma cylindrosporum was shown to be regulated by plant host and carbon sources. In the presence of a low fructose concentration, nrt2 expression could not be detected in the free-living mycelium but was high in the extramatrical symbiotic mycelium associated to the host plant Pinus pinaster. In the absence of nitrogen or in the presence of nitrate, high sugar concentrations in the medium were able to enhance nrt2 expression. Nevertheless, in the presence of high fructose concentration, high ammonium concentration still completely repressed nrt2 expression indicating that the nitrogen repression overrides sugar stimulation. This is the first report revealing an effect of host plant and of carbon sources on the expression of a fungal nitrate transporter-encoding gene. PMID- 19125304 TI - Clinical and radiological outcome of anterior-posterior fusion versus transforaminal lumbar interbody fusion for symptomatic disc degeneration: a retrospective comparative study of 133 patients. AB - Abundant data are available for direct anterior/posterior spine fusion (APF) and some for transforaminal lumbar interbody fusion (TLIF), but only few studies from one institution compares the two techniques. One-hundred and thirty-three patients were retrospectively analyzed, 68 having APF and 65 having TLIF. All patients had symptomatic disc degeneration of the lumbar spine. Only those with one or two-level surgeries were included. Clinical chart and radiologic reviews were done, fusion solidity assessed, and functional outcomes determined by pre- and postoperative SF-36 and postoperative Oswestry Disability Index (ODI), and a satisfaction questionnaire. The minimum follow-up was 24 months. The mean operating room time and hospital length of stay were less in the TLIF group. The blood loss was slightly less in the TLIF group (409 vs. 480 cc.). Intra-operative complications were higher in the APF group, mostly due to vein lacerations in the anterior retroperitoneal approach. Postoperative complications were higher in the TLIF group due to graft material extruding against the nerve root or wound drainage. The pseudarthrosis rate was statistically equal (APF 17.6% and TLIF 23.1%) and was higher than most published reports. Significant improvements were noted in both groups for the SF-36 questionnaires. The mean ODI scores at follow up were 33.5 for the APF and 39.5 for the TLIF group. The patient satisfaction rate was equal for the two groups. PMID- 19125305 TI - Fate of chlortetracycline- and tylosin-resistant bacteria in an aerobic thermophilic sequencing batch reactor treating swine waste. AB - Antibiotics have been added to animal feed for decades. Consequently, food animals and their wastes constitute a reservoir of antibiotic-resistant bacteria. The objective of this work was to characterize the impact of an aerobic thermophilic biotreatment on aerobic, antibiotic-resistant bacteria in swine waste. The proportion of tylosin- and chlortetracycline-resistant bacteria grown at 25 degrees C, 37 degrees C, and 60 degrees C decreased after treatment, but they were still abundant (10(2) to 10(8) most probable number ml(-1)) in the treated swine waste. The presence of 14 genes conferring resistance to tylosin and chlortetracycline was assessed by polymerase chain reaction in bacterial populations grown at 25 degrees C, 37 degrees C, and 60 degrees C, with or without antibiotics. In 22 cases, genes were detected before but not after treatment. The overall gene diversity was wider before [tet(BLMOSY), erm(AB)] than after [tet(LMOS), erm(B)] treatment. Analysis by denaturing gradient gel electrophoresis of amplified 16S ribosomal DNA (rDNA) fragments generally showed a reduction of the bacterial diversity, except for total populations grown at 60 degrees C and for tylosin-resistant populations grown at 37 degrees C. The latter were further investigated by cloning and sequencing their 16S rDNA. Phylotypes found before treatment were all closely related to Enterococcus hirae, whereas six different phylotypes, related to Pseudomonas, Alcaligenes, and Pusillimonas, were found after treatment. This work demonstrated that the aerobic thermophilic biotreatment cannot be considered as a means for preventing the dissemination of aerobic antibiotic-resistant bacteria and their resistance genes to the environment. However, since pathogens do not survive the biotreatment, the effluent does not represent an immediate threat to animal or human health. PMID- 19125306 TI - Fungal phyllosphere communities are altered by indirect interactions among trophic levels. AB - Trophic interactions involving predators, herbivores, and plants have been described in terrestrial systems. However, there is almost no information on the effect of trophic interactions on microbial phyllosphere community abundance, diversity, or structure. In this study, the interaction between a parasitoid, an insect herbivore, and the fungal phyllosphere community is examined. Parasitoid wasps have an indirect negative impact on fungal community diversity. On the citrus phyllosphere, the exotic wasp species, Amitus hesperidum and Encarsia opulenta, may parasitize the citrus blackfly (Aleurocanthus woglumi). If parasitism levels are low, the blackfly may produce significant amounts of honeydew secretions on the surface of the leaf. Honeydew deposition provides a carbon-rich substrate for the development of fungal growth persisting as sooty mold on the leaves. Leaves from sooty mold-infested grapefruit (Citrus paradisi) trees were collected from multiple orchards in south Texas. The effect of different levels of exotic parasite activity, citrus blackfly, and sooty mold infestation on phyllosphere mycobiota community structure and diversity was examined. Our results suggest the presence of the parasitoid may lead to a top down trophic cascade affecting phyllosphere fungal community diversity and structure. Additionally, persistent sooty mold deposits that have classically been referred to as Capnodium citri (and related asexual morphological forms) actually comprise a myriad of fungal species including many saprophytes and potential fruit and foliar pathogens of citrus. PMID- 19125307 TI - Is endoscopic band ligation superior to injection therapy for Dieulafoy lesion? AB - BACKGROUND: Dieulafoy lesion is a rare but serious cause of gastrointestinal system bleeding. An aberrant submucosal artery, which was described in 1884, causes the bleeding. The lesion can be located anywhere in the gastrointestinal tract but is most commonly found in the proximal stomach up to 6 cm from the gastroesophageal junction. Increased experience in endoscopy has led to an increased frequency of its proper diagnosis. Various methods are used to achieve successful hemostasis by endoscopy in Dieulafoy lesion; however, comparative studies about the success rates of these methods are still needed. In this study, we compared two of these endoscopic hemostatic methods: band ligation, and injection therapy in Dieulafoy lesions. METHODS: In this prospective study, 18 patients admitted to the Emergency Surgical Unit between January 2002 and December 2005 with upper gastrointestinal bleeding diagnosed as Dieulafoy lesion were included. Diagnose of Dieulafoy lesion was made at initial or second-look endoscopy. Patients were randomized in two groups according to therapy method: injection therapy and band ligation groups. Therapy was applied immediately after recognizing the lesion at the same endoscopic procedure. Two groups were compared regarding demographical data, presence of comorbid diseases, history of medication and previous gastrointestinal system bleeding, hemodynamic status, laboratory values, need for transfusion, endoscopic findings, success rate of the treatment method, mean hospital stay, complications, and recurrence of bleeding. RESULTS: Of 588 patients admitted with upper gastrointestinal hemorrhage, Dieulafoy lesion was recognized in 18 cases (3.1%) at initial or second-look endoscopy. All patients were men with a mean age of 62.8 (range, 30-80) years. Band ligation was applied to ten patients and the remaining eight were treated by injection therapy. During the follow-up period, rebleeding occurred in six of the patients (75%) with injection therapy, whereas no rebleeding occurred for the patients in the band ligation group. The rebleeding rate and mean hospital stay was significantly higher for the injection therapy group. CONCLUSIONS: Our study suggests that of the endoscopic treatment methods, band ligation is superior to injection therapy for the treatment Dieulafoy lesions. PMID- 19125309 TI - Pancreaticoduodenectomy after a Roux-en-Y gastric bypass. AB - BACKGROUND: The surgical management of periampullary lesions, status post-Roux-en Y gastric bypass procedure (RYGBP), poses a challenge. The strategy should focus on managing the gastric remnant. METHODS: We propose a technique of managing the gastric remnant while doing a pancreaticoduodenectomy (PD) in a patient with a previous RYGBP. From September 2005 to June 2008, two patients with a previous RYGBP underwent PD with a modified technique. The records were reviewed with respect to preoperative, intraoperative, and postoperative data. RESULTS: Both patients were operated for a carcinoma of the head of pancreas. Neither patient underwent a preoperative endoscopic ultrasound. The operating times were 315 and 218 min. There was no mortality or morbidity seen. Neither patient was re operated. The mean length of stay was 6 days. CONCLUSIONS: The technique suggests an approach of managing the gastric remnant and preventing delayed gastric emptying which resulted in a decreased length of hospital stay. PMID- 19125310 TI - Vascular dysfunction measured by fingertip thermal monitoring is associated with the extent of myocardial perfusion defect. AB - BACKGROUND: Previous studies have shown that vascular dysfunction measured by digital thermal monitoring (DTM) during an arm-cuff reactive hyperemia procedure correlates with the severity of coronary artery disease measured by coronary artery calcium in asymptomatic patients. Current study investigates the correlation between DTM and abnormal myocardial perfusion imaging (MPI). METHODS: About 116 consecutive patients with chest discomfort, age 57 +/- 10 years, underwent MPI, DTM and Framingham Risk Score (FRS) assessment. Fingertip temperature rebound (TR), DTM index of vascular reactivity, was assessed after a 2-minute arm-cuff reactive hyperemia test. The extent of myocardial perfusion defect was measured by summed stress score (SSS). RESULTS: TR decreased from SSS < 4 (1.61 +/- 0.15) to 4 < or = SSS < or = 8 (0.5 +/- 0.22) to 9 < or = SSS < or = 13 (0.26 +/- 0.15) to SSS > 13 (-0.37 +/- 0.19) (P = .0001). After adjusting for cardiac risk factors, the odds ratio of the lowest versus two upper tertiles of TR was 3.93 for SSS > or = 4 and 9.65 for SSS > or = 8 compared to SSS < 4. TR correlated well with SSS (r = -0.88, P = .0001). Addition of TR to FRS increased the area under the ROC curve to predict abnormal MPI, SSS > or = 4, from 0.65 to 0.84 (P < .05). CONCLUSION: Vascular dysfunction measured by DTM is associated with the extent of myocardial perfusion defect independent of age, gender, and cardiac risk factors. PMID- 19125308 TI - Clinical application of laparoscopic bariatric surgery: an evidence-based review. AB - BACKGROUND: Approximately one-third of U.S. adults are obese. Current evidence suggests that surgical therapies offer the morbidly obese the best hope for substantial and sustainable weight loss, with a resultant reduction in morbidity and mortality. Minimally invasive methods have altered the demand for bariatric procedures. However, no evidence-based clinical reviews yet exist to guide patients and surgeons in selecting the bariatric operation most applicable to a given situation. METHODS: This evidenced-based review is presented in conjunction with a clinical practice guideline developed by the Society of American Gastrointestinal and Endoscopic Surgeons (SAGES). References were reviewed by the authors and graded as to the level of evidence. Recommendations were developed and qualified by the level of supporting evidence available at the time of the associated SAGES guideline publication. The guideline also was reviewed and co endorsed by the American Society for Metabolic and Bariatric Surgery. RESULTS: Bariatric surgery is the most effective treatment for severe obesity, producing durable weight loss, improvement of comorbid conditions, and longer life. Patient selection algorithms should favor individual risk-benefit considerations over traditional anthropometric and demographic limits. Bariatric care should be delivered within credentialed multidisciplinary systems. Roux-en-Y gastric bypass (RGB), adjustable gastric banding (AGB), and biliopancreatic diversion with duodenal switch (BPD + DS) are validated procedures that may be performed laparoscopically. Laparoscopic sleeve gastrectomy (LSG) also is a promising procedure. Comparative data find that procedures with more dramatic clinical benefits carry greater risks, and those offering greater safety and flexibility are associated with less reliable efficacy. CONCLUSIONS: Laparoscopic RGB, AGB, BPD + DS, and primary LSG have been proved effective. Currently, the choice of operation should be driven by patient and surgeon preferences, as well as by considerations regarding the relative importance placed on discrete outcomes. PMID- 19125313 TI - Immunosuppressive activity of mesenchymal stem cells is not decreased in children with aplastic anemia. PMID- 19125311 TI - Chromium (VI) induced changes in growth and root plasma membrane redox activities in pea plants. AB - The effect of chromium (Cr) on growth as well as root plasma membrane redox reactions and superoxide radical production was studied in pea (Pisum sativum L. cv. Azad) plants exposed for 7 days to 20 and 200 microM Cr (VI), respectively, supplied as potassium dichromate. The growth of pea plants declined significantly at 200 microM Cr, as indicated by reduced leaf area and biomass. Relative to the control plants (no Cr exposure), the Cr content of roots increased significantly, both at 20 and 200 microM Cr. Following exposure to 200 microM Cr, there was a significant increase in root lipid peroxidation and hydrogen peroxide (H(2)O(2)) content, while both the Fv/Fm ratio and chlorophyll content were reduced. Exposure to Cr increased NADPH-dependent superoxide production in pea root plasma membrane vesicles, with the effect being more significant at 200 microM Cr than at 20 microM Cr. Treatment with Cr rapidly increased the activities of NADPH oxidase: relative to the controls, plants exposed to 20 microM Cr showed approximately a 67% increase in activity while there was a threefold increase in those plants exposed to 200 microM Cr. NADH-ferricyanide oxido-reductase activity was found to be inhibited by 16 and 51% at 20 and 200 microM Cr, respectively. The results of this study suggest that exposure to excess Cr damages pea root plasma membrane structure and function, resulting in decreased photosynthesis and poor plant growth. PMID- 19125314 TI - Middle cerebral artery occlusion 25 years after cranial radiation therapy in acromegaly: a case report. AB - We report a case of ischemic stroke in a 43 year-old woman with no traditional cardiovascular risk factors and a history of cranial surgery and cranial radiation therapy (CRT) for a GH-secreting pituitary macroadenoma. The neurological work-up on this patient disclosed several cerebral ischemic lesions and demonstrated the occlusion of the right middle cerebral artery together with the narrowing of the right carotid artery; post-radiation brain damage was also visible by nuclear magnetic resonance. We postulate the existence in this patient of a radiation-induced vascular damage, which is a well recognized process thoroughly described in in vitro studies. We remark that life-long follow-up of acromegalic patients receiving CRT is essential so that early diagnosis of radiation-induced vascular injury can be made. PMID- 19125315 TI - Yeast oxysterol-binding proteins: sterol transporters or regulators of cell polarization? AB - Oxysterol-binding protein (OSBP) and OSBP-related proteins (ORPs) are a conserved family of soluble cytoplasmic proteins that can bind sterols, translocate between membrane compartments, and affect sterol trafficking. These properties make ORPs attractive candidates for lipid transfer proteins (LTPs) that directly mediate nonvesicular sterol transfer to the plasma membrane. To test whether yeast ORPs (the Osh proteins) are sterol LTPs, we studied endoplasmic reticulum (ER)-to plasma membrane (PM) sterol transport in OSH deletion mutants lacking one, several, or all Osh proteins. In conditional OSH mutants, ER-PM ergosterol transport slowed approximately 20-fold compared with cells expressing a full complement of Osh proteins. Although this initial finding suggested that Osh proteins act as sterol LTPs, the situation is far more complex. Osh proteins have established roles in Rho small GTPase signaling. Osh proteins reinforce cell polarization and they specifically affect the localization of proteins involved in polarized cell growth such as septins, and the GTPases Cdc42p, Rho1p, and Sec4p. In addition, Osh proteins are required for a specific pathway of polarized secretion to sites of membrane growth, suggesting that this is how Osh proteins affect Cdc42p- and Rho1p-dependent polarization. Our findings suggest that Osh proteins integrate sterol trafficking and sterol-dependent cell signaling with the control of cell polarization. PMID- 19125316 TI - I-FABP expression alters the intracellular distribution of the BODIPY C16 fatty acid analog. AB - To investigate the structure-function relationships of intestinal fatty acid binding protein (I-FABP) in cellular fatty acid (FA) trafficking, we compared the distribution of a fluorescent FA analog (BODIPY FL C16) in Cos-1 cells transiently transfected with the wild type protein (wt I-FABP) to that of a variant deleted of the alpha helical domain (HL I-FABP). In vector-only cells, BODIPY fluorescence was distributed throughout the cytoplasm. In the absence of added FA, wt I-FABP was found largely in the perinuclear region with some cytoplasmic staining as well. Addition of BODIPY FL C16 to transfected cells showed that the fluorescent FA was essentially completely colocalized with the protein in the cytoplasmic and perinuclear regions as well as in cytoplasmic clusters that are not observed in the absence of wt I-FABP. For HL I-FABP, the distribution of the protein in the absence of FA was diffusely cytoplasmic, in marked contrast to the wt protein. Addition of BODIPY led to less extensive colocalization than that observed for wt I-FABP. In particular, no localization to the perinuclear region was found. Organelle colocalization studies showed that both proteins colocalized with mitochondria and endoplasmic reticulum/golgi markers, but little with a lysosomal marker. The perinuclear localization for wt I-FABP and BODIPY did not show colocalization with any of the markers tested. Taken together, these results indicate that I-FABP binds FA in vivo and that the helical domain may be important for targeting I-FABP to a perinuclear domain but not, perhaps, to the endoplasmic reticulum, golgi apparatus or mitochondria. PMID- 19125317 TI - Native LDL-cholesterol mediated monocyte adhesion molecule overexpression is blocked by simvastatin. AB - AIM OF THE STUDY: This study sought to evaluate the effect of nLDL concentrations on monocyte adhesion molecule expression in hypercholesterolemic patients with stable coronary artery disease (CAD) and to determine whether lipid-lowering therapy with simvastatin would change this effect. METHODS: Blood samples from patients with hypercholesterolemia (mean LDL 152 mg/dL) and CAD (HC, n = 23) were collected before and after a 12-week treatment with 40 mg of simvastatin. Healthy individuals (mean LDL 111 mg/dL) were used as controls (CT, n = 15). Isolated nLDL, at a fixed concentration of 100 mg/dL, was added to monocyte suspensions obtained before and after the simvastatin treatment. Monocyte activation was determined by changes in cellular adhesion molecule expression. RESULTS: In response to nLDL, CD11b and CD14 adhesion molecule expression was higher in HC patients than in CT patients before treatment (174.2 +/- 8.4 vs 102.2 +/- 6.3, P < 0.03 and 140.4 +/- 5.0 vs 90.4 +/- 6.7, P < 0.04). After simvastatin treatment, CD11b expression decreased to 116.9 +/- 12.5 (P < 0.03) and CD14 expression to 107.5 +/- 6.2 (P < 0.04). Alternatively, L-selectin expression was lower in HC patients than in CT patients before therapy (46.0 +/- 3.5 vs 62.1 +/- 5.5, P < 0.04), and it increased markedly after lipid reduction to 58.7 +/- 5.0 (P < 0.04 vs baseline). After simvastatin treatment, LDL was reduced to mean 101.5 mg/dL. CONCLUSIONS: These data demonstrate that monocytes from HC patients are more prone to marked nLDL-mediated changes of adhesion molecule expression than monocytes from controls. Simvastatin is capable of inhibiting such nLDL effects. This proinflammatory response to nLDL may have a role in the early onset of atherosclerosis. PMID- 19125318 TI - Balance between noise and adaptation in competition models of perceptual bistability. AB - Perceptual bistability occurs when a physical stimulus gives rise to two distinct interpretations that alternate irregularly. Noise and adaptation processes are two possible mechanisms for switching in neuronal competition models that describe the alternating behaviors. Either of these processes, if strong enough, could alone cause the alternations in dominance. We examined their relative role in producing alternations by studying models where by smoothly varying the parameters, one can change the rhythmogenesis mechanism from being adaptation driven to noise-driven. In consideration of the experimental constraints on the statistics of the alternations (mean and shape of the dominance duration distribution and correlations between successive durations) we ask whether we can rule out one of the mechanisms. We conclude that in order to comply with the observed mean of the dominance durations and their coefficient of variation, the models must operate within a balance between the noise and adaptation strength both mechanisms are involved in producing alternations, in such a way that the system operates near the boundary between being adaptation-driven and noise driven. PMID- 19125319 TI - Rana grylio virus thymidine kinase gene: an early gene of iridovirus encoding for a cytoplasmic protein. AB - The presence of thymidine kinase (TK) is a feature of many large DNA viruses. Here, a TK gene homologue was cloned and characterized from Rana grylio virus (RGV), a member of family Iridoviridae. RGV TK encodes a protein of 195 aa with a predicted molecular mass of 22.1 kDa. Homologues of the protein were present in all the currently sequenced iridoviruses, and phylogenetic analysis showed that it was much close to cellular TK type 2 (TK2), deoxycytidine kinase (dCK) and deoxyguanosine kinase (dGK). Subsequently, Western blotting revealed TK expression increased with time from 6 h post-infection in RGV-infected cells. Using drug inhibition analysis by protein synthesis inhibitor (cycloheximide) and DNA replication inhibitor (cytosine arabinofuranoside), RGV TK was classified as the early expression gene during in vitro infection. Subcellular localization by TK-GFP fusion protein expression and immunofluorescence staining showed RGV TK was an exclusively cytoplasmic protein in fish cells. Collectively, current data indicate that RGV TK was an early gene of iridovirus which encoded a cytoplasmic protein in fish cells. PMID- 19125320 TI - The selection pressure analysis of chicken anemia virus structural protein gene VP1. AB - Chicken anemia virus (CAV) is the pathogen of chicken infectious anemia. To clarify the driving force in CAV evolution, we have detected positive selection in the structural protein gene VP1 by using maximum-likelihood models. Strong evidence was found that VP1 proteins were subject to the high rates of positive selection, and eight sites were identified to be under positive selection using the Bayes Empirical Bayesian method. Interestingly, four selected sites (amino acids 75, 125, 141, and 144) might be responsible for the attenuation exhibited. One selected site (amino acid 287) was connected with the virulence of CAV. This study provided some implication for the evolution of CAV, development of vaccines, and investigation into the structural and functional profiles of the VP1 protein. PMID- 19125322 TI - Digital image analysis of breast epithelial cells collected by random periareolar fine-needle aspirates (RPFNA) from women at high risk for breast cancer taking hormone replacement and the aromatase inhibitor, letrozole, for six months. AB - Aromatase inhibitors are currently being evaluated as preventive agents in post menopausal women at high risk for breast cancer. A phase II trial of 42 women on hormone replacement therapy (HRT) treated with letrozole for 6 months showed Ki 67 was reduced by 66% but showed no change in cytomorphology or Masood score. Subsequent image analytical procedures (karyometry) conducted on a subset of the samples captured subvisual information that showed reduced cellular abnormality after 6 months of letrozole. In the present study we expanded on the preliminary karyometry study to determine if the change in karyometric measurements corresponded to changes in risk biomarkers quantified in the Phase II trial; and secondly, whether these biomarkers might be used together to serve as markers of response in individual cases. Pap stained slides from the Phase II trial were used. Epithelial cell images were digitized on a CCD video-microphotometer and the nuclei were segmented from the field using a semiautomatic algorithm. Nine out of 37 cases analyzed showed a numerical decrease in all three markers, although only three of these exhibited changes substantial enough to be considered as an improvement. However, 12 cases showed improvement by cytology (a decrease in Masood score of at least 2), an additional 13 cases demonstrated a reduction in Ki-67 expression by 50% of the median baseline value, and an additional five cases exhibited a decrease of at least 10% in abnormal cells by nuclear morphometry. Thus, a total of 30 of 37 cases (81%) showed improvement in at least one marker. There was no correlation between changes in Ki-67%, karyometric abnormality, and Masood score change other than specimens that exhibited an improvement in cytology also displayed greater decreases in nuclear morphometry abnormalities. Given the heterogeneity of mechanisms leading to malignancy, the quantitative analysis of nuclear chromatin patterns may be valuable as a global, or integrating, biomarker of change in chemoprevention studies in conjunction with additional markers. Correlation with long term clinical outcome is needed to validate meaningful combinations of informative biomarkers. PMID- 19125321 TI - Affective correlates of stimulant use and adherence to anti-retroviral therapy among HIV-positive methamphetamine users. AB - The use of stimulants has important implications for HIV prevention and care. However, few investigations have examined psychological correlates of substance use and adherence to anti-retroviral therapy (ART) among HIV-positive stimulant users. This cross-sectional investigation examined affective correlates of stimulant use and ART adherence among HIV-positive methamphetamine users. In total, 122 HIV-positive men who have sex with men or transgendered individuals on ART who reported using methamphetamine in the past 30 days were recruited from the community. HIV-specific traumatic stress was consistently and independently associated with more frequent cocaine/crack use (but not with methamphetamine use). Positive affect was independently associated with a decreased likelihood of reporting any injection drug use and an increased likelihood of reporting perfect ART adherence. HIV-specific traumatic stress may be an important determinant of increased cocaine/crack use in this population. Positive affect may increase the likelihood that individuals will refrain from injection drug use and achieve high levels of ART adherence. PMID- 19125326 TI - The potential premium range of risk-rating in competitive markets for supplementary health insurance. AB - In this paper, we simulate several scenarios of the potential premium range for voluntary (supplementary) health insurance, covering benefits which might be excluded from mandatory health insurance (MI). Our findings show that, by adding risk-factors, the minimum premium decreases and the maximum increases. The magnitude of the premium range is especially substantial for benefits such as medical devices and drugs. When removing benefits from MI policymakers should be aware of the implications for the potential reduction of affordability of voluntary health insurance coverage in a competitive market. PMID- 19125327 TI - Cardiac hypertrophy and thyroid hormone signaling. AB - Thyroid hormone exerts a large number of influences on the cardiovascular system. Increased thyroid hormone action increases the force and speed of systolic contraction and the speed of diastolic relaxation and these are largely beneficial effects. Furthermore, thyroid hormone has marked electrophysiological effects increasing heart rate and the propensity for atrial fibrillation and these effects are largely mal-adaptive. In addition, thyroid hormone markedly increases cardiac angiogenesis and decreases vascular tone. These multiple thyroid hormone effects are largely mediated by the action of nuclear based thyroid hormone receptors (TR) the thyroid hormone receptor alpha and beta. TRalpha is the predominant isoform in the heart. Rapid nongenomic thyroid hormone effects also occur, which can be clearly demonstrated in ex-vivo experiments. Some of the most marked thyroid hormone effects in cardiac myocytes involve influences on calcium flux, with thyroid hormone promoting expression of the gene encoding the calcium pump of the sarcoplasmic reticulum (SERCa2). In contrast, in hypothyroid animals phospholamban levels, which inhibit the SERCa2 pump, are increased. In addition, marked effects are exerted on the calcium channel of the sarcoplasmic reticulum the ryanodine channel. Related to myofibrillar proteins, myosin heavy chain alpha is increased by T3 and MHC beta is decreased. Complex and interesting interactions occur between cardiac hypertrophy induced by excess thyroid hormone action and cardiac hypertrophy occurring with heart failure. The thyroid hormone mediated cardiac hypertrophy in its initial phases presents a physiological hypertrophy with increases in SERCa2 levels and decreased expression of MHC beta. In contrast, pressure overload induced heart failure leads to a "pathological" cardiac hypertrophy which is largely mediated by activation of the calcineurin system and the MAPkinases signaling system. Recent evidence indicates that heart failure can lead to a downregulation of the thyroid hormone signaling system in the heart. In the failing heart, decreases of thyroid hormone receptor levels occur. In addition, serum levels of T4 and T3 are decreased with heart failure in the frame of the non-thyroidal illness syndrome. The decrease in T3 serves as an indicator for a bad prognosis in the heart failure patient being linked to increased mortality. In animal models, it can be shown that in pressure overload-induced cardiac hypertrophy a decrease of thyroid hormone receptor levels occurs. Cardiac function can be improved by increasing expression of thyroid hormone receptors mediated by adeno-associated virus based gene transfer. The failing heart may develop a "hypothyroid" status contributing to diminished cardiac contractile function. PMID- 19125328 TI - Natural antioxidants protect neurons in Alzheimer's disease and Parkinson's disease. AB - "Modern" medicine and pharmacology require an effective medical drug with a single compound for a specific disease. This seams very scientific but usually has unavoidable side effects. For example, the chemical therapy to cancer can totally damage the immunological ability of the patient leading to death early than non-treatment. On the other hand, natural antioxidant drugs not only can cure the disease but also can enhance the immunological ability of the patient leading to healthier though they usually have several compounds or a mixture. For the degenerative disease such as Alzheimer's disease (AD) and Parkinson's disease (PD), natural antioxidant drugs are suitable drugs, because the pathogenesis of these diseases is complex with many targets and pathways. These effects are more evidence when the clinic trial is for long term treatment. The author reviews the studies on the protecting effects of natural antioxidants on neurons in neurodegenerative diseases, especially summarized the results about protective effect of green tea polyphenols on neurons against apoptosis of cellular and animal PD models, and of genestine and nicotine on neurons against A beta-induced apoptosis of hippocampal neuronal and transgenic mouse AD models. PMID- 19125329 TI - The expression patterns of Nogo-A and NgR in the neonatal rat visual nervous system. AB - Previous studies have shown that Nogo-NgR system plays multiple roles in controlling axonal regeneration and neuronal plasticity. However, the investigation on the distributions of Nogo and NgR was limited to restricted areas or special ages, and their expression patterns in the neonatal visual nervous system is still unknown. Frozen sections were made from eyeballs and brains of three neonatal rats, and expression of Nogo and NgR proteins were examined by immunofluorescence under a laser confocal microscopy. Nogo and NgR were observed to express in the cell bodies of retina, lateral geniculate bodies and visual cortex of the neonatal rats. Our results provide evidences that the Nogo-NgR system may play an active role in the plasticity of neonatal visual system. PMID- 19125331 TI - Pressure and distortion regulate human mesenchymal stem cell gene expression. AB - While the concept that physical forces such as tension and compression are involved in mature tissue modeling is widely accepted, the role of these specific types of mechanical loading in the differentiation and maturation of uncommitted cell types like human mesenchymal stem cells (hMSCs) is currently unknown. We observed that hMSCs have the fundamental ability to distinguish between dynamic tensile and compressive loading by regulating distinct gene expression patterns and that these differences in gene expression can be related to conformational changes in cell shape and volume. Dynamic tension was found to regulate both fibroblastic and osteogenic associated genes while dynamic compression up regulated genes associated with chondrogenesis. Identifying genes involved in the mechanotransduction of different modes of physical loading in hMSC may greatly enhance the ability to rationally design tissue regeneration systems to restore proper tissue function. PMID- 19125330 TI - The serotonin transporter polymorphism rs25531 is associated with irritable bowel syndrome. AB - Irritable bowel syndrome is a frequent gastrointestinal disorder of unknown etiology. The serotonin transporter regulates the intensity and duration of serotonin signaling in the gut and is, therefore, an attractive candidate gene for irritable bowel syndrome. Previous studies investigating the 5-HTTLPR and Stin2 VNTR polymorphisms of the serotonin transporter have proved inconclusive. In this exploratory study we therefore expanded the search for a possible association of the serotonin transporter with irritable bowel syndrome to include not only the 5-HTTLPR and Stin2 VNTR length polymorphisms, but also the functional single nucleotide polymorphism rs25531. We genotyped 186 patients with irritable bowel syndrome and 50 healthy control subjects raging in age from 18 to 70 years. Carriers of the rare G allele of rs25531 had approximately threefold increased odds of irritable bowel syndrome compared with healthy controls (OR 3.3, 95% CI 1.1-9.6). Our findings suggest that further investigation of the possible role of the serotonin transporter in the etiology of IBS is warranted. PMID- 19125332 TI - Device to provide intuitive assistance in laparoscope holding. AB - This work proposes a new navigation device which autonomously handles the laparoscope, with a view to reducing latency and adjusting optics in real time. The system designed is an intuitive mechatronic device with three degrees of freedom and a sole active articulation. The device uses the point of insertion as the invariant point for navigation and has a work space that closely resembles an inverted cone. PMID- 19125333 TI - Homeostasis of brain dynamics in epilepsy: a feedback control systems perspective of seizures. AB - In an effort to understand basic functional mechanisms that can produce epileptic seizures, some key features are introduced in coupled lumped-parameter neural population models that produce "seizure"-like events and dynamics similar to the ones during the route of the epileptic brain towards seizures. In these models, modified from existing ones in the literature, internal feedback mechanisms are incorporated to maintain the normal low level of synchronous behavior in the presence of coupling variations. While the internal feedback is developed using basic feedback systems principles, it is also functionally equivalent to actual neurophysiological mechanisms such as homeostasis that act to maintain normal activity in neural systems that are subject to extrinsic and intrinsic perturbations. Here it is hypothesized that a plausible cause of seizures is a pathology in the internal feedback action; normal internal feedback quickly regulates an abnormally high coupling between the neural populations, whereas pathological internal feedback can lead to "seizure"-like high amplitude oscillations. Several external seizure-control paradigms, that act to achieve the operational objective of maintaining normal levels of synchronous behavior, are also developed and tested in this paper. In particular, closed-loop "modulating" control with predefined stimuli, and closed-loop feedback decoupling control are considered. Among these, feedback decoupling control is the consistently successful and robust seizure-control strategy. The proposed model and remedies are consistent with a variety of recent observations in the human and animal epileptic brain, and with theories from nonlinear systems, adaptive systems, optimization, and neurophysiology. The results from the analysis of these models have two key implications, namely, developing a basic theory for epilepsy and other brain disorders, and the development of a robust seizure-control device through electrical stimulation and/or drug intervention modalities. PMID- 19125335 TI - Perceived threat, risk perception, and efficacy beliefs related to SARS and other (emerging) infectious diseases: results of an international survey. AB - PURPOSE: To study the levels of perceived threat, perceived severity, perceived vulnerability, response efficacy, and self-efficacy for severe acute respiratory syndrome (SARS) and eight other diseases in five European and three Asian countries. METHOD: A computer-assisted phone survey was conducted among 3,436 respondents. The questionnaire focused on perceived threat, vulnerability, severity, response efficacy, and self-efficacy related to SARS and eight other diseases. RESULTS: Perceived threat of SARS in case of an outbreak in the country was higher than that of other diseases. Perceived vulnerability of SARS was at an intermediate level and perceived severity was high compared to other diseases. Perceived threat for SARS varied between countries in Europe and Asia with a higher perceived severity of SARS in Europe and a higher perceived vulnerability in Asia. Response efficacy and self-efficacy for SARS were higher in Asia compared to Europe. In multiple linear regression analyses, country was strongly associated with perceived threat. CONCLUSIONS: The relatively high perceived threat for SARS indicates that it is seen as a public health risk and offers a basis for communication in case of an outbreak. The strong association between perceived threat and country and different regional patterns require further research. PMID- 19125334 TI - Delivery of macromolecules using arginine-rich cell-penetrating peptides: ways to overcome endosomal entrapment. AB - Arginine-rich cell-penetrating peptides (AR-CPPs) are very promising tools for the delivery of therapeutic macromolecules such as peptides, proteins, and nucleic acids. These peptides allow efficient internalization of the linked cargos intracellularly through the endocytic pathway. However, when linked to bulky cargos, entrapment in the endocytic vesicles is a major limitation to the application of these peptides in cytosolic delivery. Attachment of a compatible endosomal escape device is, therefore, necessary to allow cytosolic delivery of the peptide-attached cargo. This review presents different endosomal escape devices currently in application in combination with AR-CPPs. Applications of fusogenic lipids, membrane-disruptive peptides, membrane-disruptive polymers, lysosomotropic agents, and photochemical internalization to enhance the cytosolic delivery of AR-CPPs-attached cargos are presented. The properties of each system and its mechanism of action for the enhancement of endosomal escape are discussed, together with its applications for the delivery of different macromolecules in vitro and, if applicable, in vivo. PMID- 19125336 TI - False heart rate feedback and the perception of heart symptoms in patients with congenital heart disease and anxiety. AB - BACKGROUND: Little is known about the mechanisms explaining an increased perception of heart symptoms in congenital heart disease (ConHD). In the present study, it was suggested that a combination of high trait anxiety and disease history increases the perception of heart symptoms. PURPOSE: It was tested whether false heart cues will result in an increased perception of heart symptoms in patients with ConHD and anxiety. METHOD: Thirty-six patients with ConHD and 44 healthy controls performed two exercise tasks. During one of the exercise tasks, participants were exposed to a false heart cue consisting of false heart rate feedback (regular or irregular). Perceived heart symptoms were assessed and heart rate, arterial partial pressure of CO(2), and respirator rate were monitored continuously. RESULTS: In line with the predictions, false heart rate feedback resulted in an increased perception of heart symptoms in high trait anxious patients with ConHD that could not be explained by acute heart dysfunction. However, unexpectedly, this effect was not observed immediately after the false heart rate feedback task but after a second exercise task without false feedback. CONCLUSION: The results suggest that not the sole presence of ConHD but ConHD in combination with high trait anxiety results in a vulnerability to overperceive heart symptoms. PMID- 19125337 TI - Characterization of promoter elements required for expression and induction by sucrose of the Arabidopsis COX5b-1 nuclear gene, encoding the zinc-binding subunit of cytochrome c oxidase. AB - Arabidopsis COX5b-1 encodes an isoform of the zinc binding subunit 5b of mitochondrial cytochrome c oxidase. A promoter region required for expression and induction by sucrose of this gene was analyzed using plants stably transformed with mutagenized promoter fragments fused to the gus reporter gene. Promoter dependent expression is absolutely dependent on a G-box present at -228 from the translation start site. This element interacts in vitro and in vivo with transcription factors from the bZip family, preferentially with the abscisic acid responsive element binding factor AREB2/ABF4. A region located upstream of the G box (-333/-259) contains elements with the core sequence ATCATT and distalB-like sequences (CCACTTG) that are required for expression in vegetative tissues. These sequences bind different sets of proteins present in plant nuclear extracts and participate in induction by sucrose (ATCATT) and abscisic acid (distalB) of the COX5b-1 promoter. We propose that the COX5b-1 promoter has acquired novel regulatory mechanisms during evolution after gene duplication. These novel mechanisms have allowed the diversification of expression patterns, but also the conservation of some responses that, as induction by sucrose, are shared by COX5b 1 and other genes encoding components of the mitochondrial respiratory chain. Conservation of these responses may be a pre-requisite for the successful incorporation of new regulatory elements in this class of genes. PMID- 19125338 TI - Groundwater chemistry and arsenic mobilization in the Holocene flood plains in south-central Bangladesh. AB - A comparative study of arsenic enrichment in the Bengal Delta (BD) was carried out in three alluvial aquifers in south-central Bangladesh. Investigated sites included Sonargaon in Narayanganj, Chandina in Comilla and Sirajdikhan in Munshiganj districts. At all sites samples from different depths were collected, and water chemistry and redox status vs. depth trends were determined. The concentrations of DOC and HCO(3)(-) were highest at Sirajdikhan site, while at the Sonargaon and Chandina sites the concentrations were lower. On the contrary, the NH(4)(+) concentration was high at the Chandina site as compared to the other sites. There was a good match between dissolved As and Fe at the Sirajdikhan and Sonargaon sites, but not at the Chandina site. The dissolved aqueous concentration of Mn was low at the Chandina site, which suggested that the Mn(IV) redox buffering step was missing. Speciation modeling indicated a possibility of siderite precipitation at all sites, but precipitation of rhodochrosite only at the Sonargaon and Sirajdikhan sites. At the Sirajdikhan site, the log P(CO2) values were very high (-1.37), which revealed the production of CO(2) in redox processes. Principal component analysis (PCA) indicated an impact of sea water and redox status of different samples. These results suggest that the dissolved As is de-coupled from dissolved Mn because when released, As is re-adsorbed onto the Fe(III) minerals in solid phase, as well as from dissolved Fe when precipitation of Fe(II) minerals controls the aqueous concentrations of Fe. In addition, several other concurrent redox processes may exert kinetic constraints depending on refractory characteristics of Fe(III) minerals. PMID- 19125339 TI - A comparative study of the extractability of arsenic species from silverbeet and amaranth vegetables. AB - There is still no reliable standard extraction method for the speciation of arsenic (As) in plant tissue, and hence there is great interest in developing one for plants that are used as human food. Speciation and bioavailability are critical for accurate human health risk assessment, as As species vary in both their toxicity and bioavailability. Recent incidences of As poisoning in many countries have led to significant research into the fate and dynamics of As in the soil and water environment, including speciation. Although one of the major pathways of ingestion of As is via food, only limited research has been conducted to assess the nature and proportion of various As species present in food crops. In this study, we compared the efficacy of ammonium dihydrogen phosphate and protein extracting solution for the extractability of As from two different species of spinach (amaranth and silverbeet). We found that a microwave-assisted technique with protein extracting solution was most effective, yielding 76-114% extractability and excellent separation and speciation of all As species present in the spinach matrices. The stability test for extracted As species showed them as stable for 45 days without any significant loss or inter-conversion. Both As(III) and As(V) were identified in the shoots of amaranth and silverbeet. However, the percentage of As species varied between amaranth and silverbeet. The silverbeet shoot showed a somewhat higher percentage of As(V), while the amaranth showed a higher percentage of As(III). The samples contained mostly inorganic As, especially As(III) (>90%) in the edible part of the vegetables, a form that is more toxic and bioavailable than other organic and methylated species. PMID- 19125341 TI - Experimental and theoretical investigation of implantable cardiac pacemaker exposed to low frequency magnetic field. AB - This paper presents in vitro investigation of an implantable cardiac pacemaker exposed to low frequency magnetic fields. The method used in this study is based on the interaction by inductive coupling through the loop formed by the pacemaker and its loads and the surrounding medium. This interaction results in an induced electromotive force between the terminals of the pacemaker, which can potentially disturb its operation. The studied frequencies are 50/60 Hz and 10/25 kHz. The experimental tests were carried out on several cardiac pacemakers, single chamber, and dual chamber. The results show a window effect of the detection circuits of cardiac pacemakers for the four studied frequencies. The modelling of the test bed requires studying the effects of the induced currents generated by the application of a magnetic field. Analytical calculations and Numerical simulations were carried out. We modelled the interactions of the magnetic field with a simplified representation of pacemaker embedded in the medium. The comparison of the results in the air and in vitro enabled us to make an equivalent electric model. The results obtained in experimental and theoretical studies allowed us to validate the test bed. The method applied is valid for other medical implants such as cardiac defibrillators, implant hearing aids system...etc. PMID- 19125340 TI - Myosin regulatory light chain phosphorylation inhibits shortening velocities of skeletal muscle fibers in the presence of the myosin inhibitor blebbistatin. AB - Phosphorylation of skeletal myosin regulatory light chain (RLC) occurs in fatigue and may play a role in the inhibition of shortening velocities observed in vivo. Forces and shortening velocities were measured in permeabilized rabbit psoas fibers with either phosphorylated or dephosphorylated RLCs and in the presence or absence of the myosin inhibitor blebbistatin. Addition of 20 microM blebbistatin decreased tensions by approximately 80% in fibers, independent of phosphorylation. In blebbistatin maximal shortening velocities (V(max)) at 30 degrees C, were decreased by 45% (3.2 +/- 0.34 vs. 5.8 +/- 0.18 lengths/s) in phosphorylated fibers but were not inhibited in dephosphorylated fibers (6.0 +/- 0.30 vs. 5.4 +/- 0.30). In the presence of 20 microM blebbistatin, K(m) for V(max) as a function of [ATP] was lower for phosphorylated fibers than for dephosphorylated fibers (50 +/- 20 vs. 330 +/- 84 microM) indicating that the apparent binding of ATP is stronger in these fibers. Phosphorylation of RLC in situ during fiber preparation or by addition of myosin light chain kinase yielded similar data. RLC phosphorylation inhibited velocity in blebbistatin at both 30 and 10 degrees C, unlike previous reports where RLC phosphorylation only affected shortening velocities at higher temperatures. PMID- 19125343 TI - Fluorous Lewis acids and phase transfer catalysts. AB - The new phase-separation and immobilization technique known as fluorous biphase system (FBS) has become an active topic among researchers in both industry and academia. A series of fluorous biphasic reactions catalyzed by Lewis acid-type metal perfluorooctanesulfonates and metal bis(perfluorooctanesulfonyl)amides are reviewed in this paper. The recent development of fluorous phase transfer catalyst (PTC) is also discussed. PMID- 19125344 TI - Changes of bladder activity and connexin 43-derived gap junctions after partial bladder-outlet obstruction in rats. AB - INTRODUCTION: We investigated changes of vesical gap junctions in relation to changes of the micturition reflex in rats with partial bladder-outlet obstruction (BOO). MATERIALS AND METHODS: A total of 66 female Sprague-Dawley rats were divided into six groups: sham operation (control); 3, 14, and 28 days after BOO; and 3 and 28 days after relief of BOO lasting for a three-day period. Under urethane anesthesia, isovolumetric cystometry was performed on each group. Expression of mRNA for the gap-junction protein connexin 43 (Cx43) in the bladder was measured in each group. Immunohistochemistry using Cx43 antibody was also performed on the bladder after BOO. RESULTS: The interval between bladder contractions was shorter in all of the other groups than in the control group. Expression of Cx43 mRNA was increased 3, 14, and 28 days after BOO (the peak increase was twofold), and three days after the relief of BOO, but it returned to the control level by 28 days after relief of BOO. Histologically, smooth muscle hypertrophy was detected in the bladder after BOO and punctate staining of the smooth muscle by Cx43 antibody increased after BOO. CONCLUSION: These results suggest that partial BOO produces detrusor overactivity that may depend on increased intercellular communication via gap junctions in the bladder. Relief of BOO led to a decrease of Cx43 mRNA, but detrusor overactivity persisted in the chronic phase, suggesting a reversible change of vesical gap junctions and an irreversible change of bladder activity after BOO. PMID- 19125345 TI - Altered P-selectin and CD44 expression in the renal tissues and peripheral blood of children with IgA nephropathy. AB - OBJECTIVE: To understand the role of P-selectin (CD62P) and CD44 in mediating immune inflammation in the nephrotic process of children with IgA nephropathy (IgAN), cooperative expression of CD62P and CD44 in peripheral blood and renal tissues of IgAN children was investigated and its association with changes of histopathologic, serologic, and urinary properties was tested. MATERIAL AND METHODS: Forty-six IgAN children were divided into three groups according to pathologic grades and clinical features. Fifteen blood samples from normal children and four normal renal biopsy specimens were used as controls. Plasma level of CD62P was detected by double antibody sandwich immunoradiometric assay; ELISA was used to determine serum level of CD44. Expression of CD62P and CD44 in renal tissues was determined by immunohistochemistry. RESULTS: Cooperative expression of CD62P and CD44 was detected in renal tissues and peripheral blood of IgAN children. Altered expression of CD62P and CD44 in peripheral blood significantly correlated not only with hematuria, proteinuria, serum cholesterol, and albumin, and with urine NAG and beta(2)-MG, but also with degree of tubulointerstitial injury in IgAN children. CONCLUSION: The evidence supported CD62P and CD44 as initial and promoting factors mediating immune inflammation in the nephrotic process in IgAN children. The cooperative expression profiles of CD62P and CD44 in renal tissues and peripheral blood combined with serologic and urinary predictors may be important in diagnosis of progression in children with IgAN. PMID- 19125346 TI - Alternative respiration of fungus Phycomyces blakesleeanus. AB - Respiratory characteristics of germinating spores, developing mycelium and mitochondria of the fungus Phycomyces blakesleeanus were investigated by means of oxygen Clark-type electrode. The effects of respiratory inhibitors and metabolic compounds on oxygen consumption were tested. It was demonstrated that P. blakesleeanus apart of cyanide-sensitive respiration, CSR, possess alternative respiration, (cyanide-resistant respiration, CRR) which is constitutive and whose capacity decreases during development. Maximum is observed for activated spores where CRR capacity is significantly greater than CSR. After treatment with antimycin A, a third type of respiration insensitive to antimycin A and low concentration of SHAM (sufficient for inhibition of CRR), but sensitive to cyanide and high concentration of SHAM, has been expressed. PMID- 19125347 TI - Light-activated cytotoxic compounds from Malaysian microorganisms for photodynamic therapy of cancer. AB - Photodynamic therapy (PDT) is a promising cancer treatment which involves activation of a photosensitizing drug with light to produce reactive oxygen species that kill tumors without causing damage to unirradiated normal tissues. To date, only Photofrin, Foscan and Levulan have been approved for clinical treatment of cancer. Tropical habitats such as those found in Malaysia are attractive sources of new therapeutic compounds as tremendous chemical diversity is found in a large number of plants, animals, marine- and micro-organisms. In our screening program for novel photosensitizers from nature, colorful strains of fungi (from Aspergillus and Penicillium genus) and bacteria (including actinomycetes and photosynthetic bacteria) were collected from various habitats in Peninsular Malaysia, such as coastal soil, peat soil, marine sponges and wastewater ponds. Methanolic extracts from a total of 85 different species were evaluated with a short-term cell viability assay for photo-cytotoxicity, where a promyelocytic leukemia cell-line, HL60 incubated with 20 microg/ml of extracts was irradiated with 9.6 J/cm(2) of a broad spectrum light. Two of these extracts, one from Rhodobacter sphaeroides (PBUM003) and one from Rhodopseudomonas palustris (PBUM001) showed moderate to strong photo-cytotoxicity. Subsequent bioassay guided isolation of the PBUM001 extract yielded known photosensitisers that are based on bacteriochlorophyll-a by comparing their molecular weight data, HPLC profiles and UV-vis absorption spectra with literature values, thereby demonstrating the validity of our screening approach. PMID- 19125348 TI - Rifamycin S and its geometric isomer produced by a newly found actinomycete, Micromonospora rifamycinica. AB - Strain AM105 was separated from mangrove sediment in the South China Sea in this research. The morphological and genomic data showed that the strain merits description as a novel species, proposed as Micromonospora rifamycinica. From the acetate ethyl extract of its fermentation broth, two antibiotics against Gram positive bacteria (including MRSA), rifamycin S and its geometric isomer were isolated. Their structures were elucidated on the basis of spectroscopic analyzes. (1)H and (13)C NMR data of the isomer of rifamycin S were first described in this paper. PMID- 19125349 TI - A viewpoint: why chlorophyll a? AB - Chlorophyll a (Chl a) serves a dual role in oxygenic photosynthesis: in light harvesting as well as in converting energy of absorbed photons to chemical energy. No other Chl is as omnipresent in oxygenic photosynthesis as is Chl a, and this is particularly true if we include Chl a(2), (=[8-vinyl]-Chl a), which occurs in Prochlorococcus, as a type of Chl a. One exception to this near universal pattern is Chl d, which is found in some cyanobacteria that live in filtered light that is enriched in wavelengths >700 nm. They trap the long wavelength electronic excitation, and convert it into chemical energy. In this Viewpoint, we have traced the possible reasons for the near ubiquity of Chl a for its use in the primary photochemistry of Photosystem II (PS II) that leads to water oxidation and of Photosystem I (PS I) that leads to ferredoxin reduction. Chl a appears to be unique and irreplaceable, particularly if global scale oxygenic photosynthesis is considered. Its uniqueness is determined by its physicochemical properties, but there is more. Other contributing factors include specially tailored protein environments, and functional compatibility with neighboring electron transporting cofactors. Thus, the same molecule, Chl a in vivo, is capable of generating a radical cation at +1 V or higher (in PS II), a radical anion at -1 V or lower (in PS I), or of being completely redox silent (in antenna holochromes). PMID- 19125350 TI - Prospective treatment of cerebrotendinous xanthomatosis with cholic acid therapy. AB - Cerebrotendinous xanthomatosis (CTX) OMIM#213700 is a rare disorder of bile acid synthesis caused by deficiency of the enzyme sterol 27-hydroxylase. It results in deficiency of bile acids and accumulation of abnormal bile alcohols and accelerated cholesterol synthesis. CTX usually presents in the second or third decade with slowly progressive neurological dysfunction, cerebellar ataxia and premature atherosclerosis. Treatment with bile acid supplementation improves but does not completely reverse the neurological signs and symptoms. However, CTX is now known to be associated with a period of neonatal cholestasis. If it is diagnosed at this point, treatment may prevent the onset of neurological problems. We present the case histories and developmental findings in two affected siblings treated from infancy. We plan to continue regular neurodevelopmental reviews. PMID- 19125352 TI - Effects of temperature and culture media on vegetative growth of an entomopathogenic fungus Isaria sp. (Hypocreales: Clavicipitaceae) naturally affecting the whitefly, Bemisia tabaci in Texas. AB - The effects of temperature and mycological media on mycelial growth and estimates of spore production of an indigenous entomopathogenic fungus, Isaria sp., found during natural epizootics on whiteflies in the Lower Rio Grande Valley of Texas, were investigated. The radial growth (mm/day) of Isaria sp. as a function of temperature fits a linear model; with faster growth on Sabouraud dextrose agar with yeast extract, SDAY slopes (0.23) than on Sabouraud maltose agar, SMA slopes (0.14) from 20 to 30 degrees C, with an optimal temperature of 30 degrees C (SDAY: 4.1 mm, SMA: 3.1 mm). Moderate growth occurred at 25 degrees C (SDAY: 3.4 mm, SMA: 2.7 mm). Growth was lowest at 20 degrees C (SDAY: 1.9 mm, SMA: 1.8 mm). No fungal growth was observed at 35 degrees C and 40 degrees C. However, when Isaria sp. was exposed to 35 degrees C for the first 7 days, it could recover and grow when transferred to 25 degrees C (SDAY: 3.5 mm, SMA: 2.8 mm). No recovery or growth occurred after transfer from 40 degrees C to 25 degrees C. The average conidial production on SDAY after 20 days incubation at 25 degrees C and a photoperiod of 14:10 h light: dark was 1.2 x 10(8) conidia/cm(2) with 100% spore viability. When compared on SDAY at 25 degrees C, the radial growth rate of I. javanica ex type CBS 134.22 (5.1 mm/day) was greater than seven Isaria isolates including Isaria sp.; but maximum growth rates were similar among all related Isaria isolates (90-97%). The Isaria sp. fungus tolerates high temperatures (35 degrees C), suggesting that it is naturally selected for the subtropical semi arid environment, where it could serve as an important natural control agent of the sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotype B, one of the most invasive and economically damaging insects to agriculture. PMID- 19125351 TI - Clinical and molecular features of mitochondrial DNA depletion syndromes. AB - Mitochondrial DNA depletion syndromes (MDSs) form a group of autosomal recessive disorders characterized by profoundly decreased mitochondrial DNA copy numbers in affected tissues. Three main clinical presentations are known: myopathic, encephalomyopathic and hepatocerebral. The first is associated with mutations in thymidine kinase 2 (TK2) and p53-induced ribonucleotide reductase B subunit (RRM2B); the second with mutations in succinate synthase A (SUCLA2) and B (SUCLG1); the third with mutations in Twinkle (PEO1), pol-gammaA (POLG1), deoxyguanosine kinase (DGUOK) and MPV17 (MPV17). In this work, we review the MDS associated phenotypes and present our own experience of 32 MDS patients, with the aim of defining the mutation frequency of the known genes, the clinical spectrum of the diseases, and the genotype-phenotype correlations. Five of our patients carried previously unreported mutations in one of the eight MDS genes. PMID- 19125353 TI - Effects of puerarin on receptor for advanced glycation end products in nephridial tissue of streptozotocin-induced diabetic rats. AB - To explore the effects of puerarin on mRNA expression of advanced glycation end products (AGE) specIfic cellular receptor (RAGE) in renal cortex of diabetic rats induced by streptozotocin (STZ). We induced Diabetic rats by an intraperitoneal injection of STZ in Sprague-Dawley (SD) rats. 30 male SD rats were randomly divided into 3 groups, diabetes adding puerarin group (DP group, n = 11, intraperitoneal injection of puerarin 100 mg/kg d), Diabetes group (D group, n = 11) and normal control group (C group, n = 8). The body weight (BW) and blood glucose (BG) were measured every 2 weeks. eight weeks later, all rats were sacrificed and the expression of RAGE mRNA was detected in renal cortex by reverse transcription-polymerase chain reaction (RT-PCR), respectively, and renal AGEs content was determined by fluorescence microscopy. Compared with those of control group, the BW and BG were lower in DP group and D group at 8th week (P < 0.01). RAGE/beta-actin ratio were 0.263 +/- 0.023, 0.435 +/- 0.010, 0.141 +/- 0.045, respectively, in DP group, D group and C group, and there was significant difference between every two groups (P < 0.01). The renal AGEs fluorescence intensity of DP group was weaker than D group, stronger than C group. Puerarin can protect the renal tissue from the impairment of hyperglycemia and AGE by decreasing AGEs contents and inhibiting of the expression of RAGE mRNA in the kidney. PMID- 19125354 TI - Work status, daily activities and quality of life among people with severe mental illness. AB - OBJECTIVE: Work is often a desired goal for people with mental illness, as expressed by both themselves and their relatives. This study investigated the importance of work status, everyday activities and objective life indices for subjective quality of life, with a special focus on quality of life domains. METHOD: The sample consisted of 103 individuals with severe mental illness in Sweden, a majority of whom had schizophrenia. Interview-based questionnaires were used to assess quality of life (MANSA) and activity factors (SDO, OVal-pd). RESULTS: Work status and activity in terms of actual doing were of some, but minor, importance to subjective quality of life domains, whereas satisfying and valuable activities were consistently associated with most quality of life domains. CONCLUSION: Although no causal relationship could be established, the findings indicate that open-market work might not be decisive for subjective quality of life, but that satisfying and meaningful everyday activities could contribute to a better life quality for those who have a severe and lasting mental illness. PMID- 19125355 TI - Differential role of the carboxy-terminus of the A(2B) adenosine receptor in stimulation of adenylate cyclase, phospholipase Cbeta, and interleukin-8. AB - In human mast cells and microvascular endothelial cells, the A(2B) adenosine receptor controls at least three independent signaling pathways, i.e., Gs mediated stimulation of adenylate cyclase, Gq-mediated stimulation of phospholipase Cbeta, and Gs/Gq-independent upregulation of IL-8. Functional analysis of cells transfected with full-length and truncated receptor constructs revealed that the A(2B) receptor C-terminus is important for coupling to Gs and Gq proteins. Removal of the entire cytoplasmic portion in the A(2B) receptor C terminus rendered it incapable of stimulating adenylate cyclase and phospholipase Cbeta. Conversely, removal of the distal 16 amino acids facilitated signal transduction from the receptor to the downstream Gs but not Gq proteins. However, the A(2B) receptor C-terminus is not essential for upregulation of IL-8. Analysis of chimeric A(2A)/A(2B) receptors demonstrated that only chimeras containing the third intracellular loop of the A(2B) receptor mediated agonist-dependent IL-8 reporter stimulation, suggesting that this domain is important for upregulation of IL-8. PMID- 19125356 TI - Modelling attention in individual cells leads to a system with realistic saccade behaviours. AB - Single cell recordings in monkey inferior temporal cortex (IT) and area V4 during visual search tasks indicate that modulation of responses by the search target object occurs in the late portion of the cell's sensory response (Chelazzi et al. in J Neurophysiol 80:2918-2940, 1998; Cereb Cortex 11:761-772, 2001) whereas attention to a spatial location influences earlier responses (Luck et al. in J Neurophysiol 77:24-42, 1997). Previous computational models have not captured differences in the latency of these attentional effects and yet the more protracted development of the object-based effect could have implications for behaviour. We present a neurodynamic biased competition model of visual attention in which we aimed to model the timecourse of spatial and object-based attention in order to simulate cellular responses and saccade onset times observed in monkey recordings. In common with other models, a top-down prefrontal signal, related to the search target, biases activity in the ventral visual stream. However, we conclude that this bias signal is more complex than modelled elsewhere: the latency of object-based effects in V4 and IT, and saccade onset, can be accurately simulated when the target object feedback bias consists of a sensory response component in addition to a mnemonic response. These attentional effects in V4 and IT cellular responses lead to a system that is able to produce search scan paths similar to those observed in monkeys and humans, with attention being guided to locations containing behaviourally relevant stimuli. This work demonstrates that accurate modelling of the timecourse of single cell responses can lead to biologically realistic behaviours being demonstrated by the system as a whole. PMID- 19125357 TI - Image manipulation as research misconduct. AB - A growing number of research misconduct cases handled by the Office of Research Integrity involve image manipulations. Manipulations may include simple image enhancements, misrepresenting an image as something different from what it is, and altering specific features of an image. Through a study of specific cases, the misconduct findings associated with image manipulation, detection methods and those likely to identify such manipulations, are discussed. This article explores sanctions imposed against guilty researchers and the factors that resulted in no misconduct finding although relevant images clearly were flawed. Although new detection tools are available for universities and journals to detect questionable images, this article explores why these tools have not been embraced. PMID- 19125358 TI - The role of long-term memory in digit-symbol test performance in young and older adults. AB - The psychological functions assessed by substitution tests, and the age-related performance decline, are not well understood. Here several aspects of long-term memory were manipulated across younger and older adults. A 45-page Digit-Symbol test was employed. Each page contained a 9-item digit symbol code-table and 9 response items. There were 9 study conditions with each condition deployed across 5 pages, or trials, of the test. The conditions were formed by crossing two within-subjects factors, each with 3 levels. The first factor, Digit Order, pertained to having the code table digits in numerical order vs. a pseudo-random order fixed across trials vs. a pseudo-random order that varied across trials. The second factor, Symbol Pairing, pertained to having a fixed digit-symbol pairing across trials vs. having a varying digit-symbol pairing across trials vs. having a novel set of 9 symbols introduced on each of the 5 trials. Including the additional factor, Age, resulted in a 2 x 3 x 3 mixed randomised block design. The older group was slowed, F(1, 22) = 17.267, p < .001, and overall-performance was poorer when the digits were arranged non-numerically, F(1,44) = 55.403, p < .001. An Age by Symbol-Order interaction indicated that use of novel symbols disadvantaged only the older participants, F(1, 44) = 6.577, p = .014. While there was no evidence that incidental paired-associate learning or spatial memory affect digit-symbol performance, symbol familiarity may be important to digit symbol test completion in older adults. The benefit of ordinally arranged digits in the coding table highlights a fundamental process difference between Digit Symbol and Symbol-Digit test formats. PMID- 19125360 TI - Psychopathological and behavior impairments in Williams-Beuren syndrome: the influence of gender, chronological age, and cognition. AB - The aim of this study was to explore psychopathological and behavior impairments in Williams-Beuren syndrome (WBS) by focusing on individual differences rather than group tendencies. Parent/Guardian ratings on the Child Behavior Checklist were analyzed in order to examine the influence of gender, chronological age, and cognitive abilities on psychopathological impairment within WBS. In line with predictions, and consistent with cognitive heterogeneity in WBS, psychopathological and behavioral abnormalities were variable, with gender and specific cognitive abilities making significant and independent contributions to this variance. For gender, females were significantly more likely than males with WBS to display difficulties with externalizing problems. For specific cognitive abilities, those cognitive functions characteristic of the classic WBS cognitive profile (a strength in verbal skills and a weakness in spatial skills) related to significantly greater internalizing difficulties. Future studies should explore underlying genetic and neurological differences in individuals with WBS in order to help explain the variability in psychopathological and cognitive functions. PMID- 19125361 TI - Measuring mindfulness: pilot studies with the Swedish versions of the Mindful Attention Awareness Scale and the Kentucky Inventory of Mindfulness Skills. AB - The present article describes data from pilot studies with the Swedish versions of the Mindful Attention Awareness Scale (MAAS) and the Kentucky Inventory of Mindfulness Skills (KIMS). The MAAS and two of the KIMS scales, Act with Awareness and Accept without Judgment, were found to correlate in the predicted direction with measures of well-being and emotional distress. The KIMS scales Observe and Describe showed more ambiguous results, possibly because these two scales measure a mixture of two opposite kinds of processes: healthy self observation ("experiential self-focus") and unhealthy rumination ("analytical self-focus"). The KIMS was also used in an uncontrolled study with participants in an educational programme for close relatives of persons with borderline personality disorder, Family Connections (FC), which includes components of mindfulness training. The FC participants were found to (a) score lower than a comparison group on Act with Awareness and Accept without Judgment before treatment and (b) show significantly increased scores on Accept without Judgment after treatment. PMID- 19125362 TI - Survey among patients with basal cell carcinoma in The Netherlands. AB - This paper describes the findings of a survey distributed among Dutch patients with basal cell carcinoma (BCC). The questionnaire comprised a list of questions related to demographic characteristics, features of BCC, reason for consulting a dermatologist, anxiety, type of treatment and the satisfaction with this treatment and desired benefits of treatment. In total, 220 patients completed the survey. The age of these responders varied between 27 and 89 years (mean 64.6 years). Half of the patient group had already previously experienced a BCC. Most patients (52%) indicated that the diagnosis 'skin cancer' frightened them, but that they knew it could be treated. Accordingly, most patients (70%) indicated that BCC had no or hardly any influence on their quality of life. From the patient's perspective, efficacy, low recurrence rate and no or minor scarring are important features of a BCC treatment. Surgery was the most popular therapy. The number of BCC patients is growing, which will lead to a definite burden for dermatologists in the near future. Our survey demonstrated that patients are mostly interested in the efficacy, low recurrence rates and cosmetic outcome of their therapies. Newly efficacious and non-invasive therapies, such as the recently introduced photodynamic therapy or home treatment with imiquimod, can help to overcome these concerns. PMID- 19125363 TI - Effects of various parameters of the 1064 nm Nd:YAG laser for the treatment of enlarged facial pores. AB - BACKGROUND: A variety of treatment modalities have been used to reduce the size of enlarged facial pores without obvious success. OBJECTIVES: To assess and compare the effects of various parameters of a 1064 nm Nd:YAG laser in the treatment of enlarged facial pores. METHODS: This was a prospective intra individual left-right comparative study. A total of 40 individuals with enlarged facial pores were recruited for this study. Ten individuals were respectively treated on one half of the face with a quasi long-pulsed 1064 nm Nd:YAG laser (method 1), a Q-switched 1064 nm Nd:YAG laser (method 2), both quasi long-pulsed and Q-switched 1064 nm Nd:YAG lasers without carbon-suspended lotion (method 3), and both quasi long-pulsed and Q-switched 1064 nm Nd:YAG lasers with carbon suspended lotion (method 4). The other half of the face was left untreated as a control. Five laser sessions were performed with a 3-week interval. The pore sizes were measured using an image analysis program and the sebum level was measured with a Sebumeter before and after the treatments. RESULTS: The pore size and sebum level decreased in all four methods on the treated side compared to the control (p < 0.05). CONCLUSIONS: Treatment with a 1064 nm Nd:YAG laser is an effective method for reducing pore size and sebum level. PMID- 19125364 TI - A comparison of the effects for sustained versus shifted attention on dichotic listening performance. AB - We measured the effect of two types of directed attention instructions, sustained by a verbal cue or shifted by a tone cue with different time intervals (150, 450, and 750 ms), on a consonant-vowel dichotic listening (C-V DL) test for a large group of right- and left-handed participants of both sexes. An increasing of the hits and a decreasing of the intrusions from the baseline DL test scores was evident for both types of attentional manipulations, with no differences regarding sex or handedness. Increasing the time from 150 to 450 ms benefited the focusing of attention but this advantage was markedly attenuated at the longer 750-ms interval. The improving effect was seen for the hits of both the left and right ears and so a right ear advantage was evident for the three time intervals. PMID- 19125365 TI - Editorial commentary: is LRRTM1 the gene for handedness? PMID- 19125366 TI - Where and what is the right shift factor or cerebral dominance gene? A critique of Francks et al. (2007). AB - Francks et al. (2007, p. 1129) claim to have identified "The first potential genetic influence on human handedness ... and the first putative genetic effect on variability in human brain asymmetry" and a gene "that underlies much of human cognition, behaviour and emotion" (p. 1129). We criticise this claim on the basis that the authors have made unjustified assumptions concerning mode of transmission both of psychosis and relative hand skill, that they have failed to establish a parent of origin effect, and have overlooked previous findings concerning the genetic basis of handedness and asymmetry. We suggest that some of these errors relate to the application of linkage disequilibrium to detect variation that is common in the population and relates to the characteristic that defines the species. While we agree (and indeed first proposed) that the variation underlying psychosis is intrinsically related to the cerebral torque, which we take to be the anatomical basis of language, we are unconvinced by the data for LRRTM1 presented by Francks et al. We consider that a stronger case can be mounted for the Protocadherin11X/Y gene pair located in the hominid specific Xq21.3/Yp11.2 region of homology that was generated by a duplication from the X between 6 and 5 million years ago and that has been subject to a number of chromosomal and sequence changes. This gene pair can account for relationships between relative hand skill and verbal and non-verbal ability that are sex dependent, and morphological changes in the brain in psychosis that reflect interactions between sex and laterality, which are already established in the literature. PMID- 19125367 TI - Understanding the genetics of behavioural and psychiatric traits will only be achieved through a realistic assessment of their complexity. AB - Francks et al. (2007) performed a recent study in which the first putative genetic effect on human handedness was identified (the imprinted locus LRRTM1 on human chromosome 2). In this issue of Laterality, Tim Crow and colleagues present a critique of that study. The present paper presents a personal response to that critique which argues that Francks et al. (2007) published a substantial body of evidence implicating LRRTM1 in handedness and schizophrenia. Progress will now be achieved by others trying to validate, refute, or extend those findings, rather than by further armchair discussion. PMID- 19125368 TI - Erythrocyte and reticulocyte indices in iron deficiency in chronic kidney disease: comparison of two methods. AB - OBJECTIVE: Anaemia is a common complication of chronic kidney disease (CKD), particularly in dialysis patients. The recent European guidelines for anaemia treatment in CKD indicate the percentage of hypochromic red cells (%HYPO) and reticulocyte haemoglobin content (CHr) calculated by Siemens ADVIA haematology analysers as a useful tool indicating iron deficiency. The aim of this study was to evaluate the agreement between CHr and %HYPO parameters and the reticulocyte haemoglobin equivalent (RET-He) and red blood cell haemoglobin equivalent (RBC He) calculated by the Sysmex XE-2100 haematology analyser in a cohort of 200 dialysis patients referred to the Nephrology Unit of our hospital. Furthermore, we evaluated a new index, the DF-Hypo XE, obtained from haemoglobin (Hb), haematocrit (Hct) and RET-He, provided by the Sysmex XE-2100, as a new potential marker of %HYPO in dialysed patients. MATERIAL AND METHODS: Blood samples collected in EDTA anticoagulant from 200 CKD patients receiving erythropoietin and iron to maintain haemoglobin level between 10 and 12 mg/dL were analysed on both the Siemens ADVIA 2120 and the Sysmex XE-2100 within 2 h of collection. RESULTS: There was good correlation between CHr and RET-He (r = 0.88; p<0.0001), %HYPO and DF-Hypo XE (r = 0.89; p<0.0001) and between RBC-He and CH (r = 0.96; p<0.0001), but there was a lower correlation, even though statistically significant, between RBC-He and %HYPO (r = -0.59; p<0.0001). The Altman-Bland analysis showed a very good level of agreement between CHr and RET-He (mean bias = 1.04 pg), %HYPO and DF-Hypo XE (mean bias = 1.73). Using a cut-off value of 29.4 pg for the RET-He and of 10.2 for the DF-Hypo XE, 15 out 17 patients with a CHr <29.0 pg and 9 out 11 patients with a %Hypo <10.0% were respectively correctly identified. CONCLUSIONS: Our study shows good correlation and agreement between CHr and RET-He and between %HYPO and DF-Hypo XE in evaluating CKD patients needing iron support. PMID- 19125369 TI - Sex determines the influence of smoking and gene polymorphism on glutathione peroxidase activity in erythrocytes. AB - OBJECTIVE: Glutathione peroxidase 1 (GPX1) is one of the major oxidative enzymes. Our aim was to characterize factors influencing its activity and to determine whether or not the activity is associated with asthma. MATERIAL AND METHODS: Serum selenium concentration was measured, GPX1 polymorphisms were genotyped and smoking history was obtained in a Danish population-derived case-base cohort of 1,191 subjects designed to evaluate risk factors for asthma. GPX1 activity was measured in 134 male and 164 female subjects equally distributed according to genotype of GPX1. Among these subjects, 82 (28 %) had doctor-diagnosed asthma. RESULTS: The average serum selenium concentration was too low for optimal enzyme activity (mean (SE), 83.4 (0.76) ng/mL). GPX1 activity in men was lower than in women, 52.6 (0.66) and 56.4 (0.59) U/g protein, respectively (p<0.001). In men, activity was positively associated with serum selenium concentration (p = 0.005) and negatively associated with both active smoking (p = 0.009) and exposure to environmental tobacco smoke (p = 0.02). In women, activity was associated with genotypes with 59.2 (1.4), 56.0 (1.4) and 54.2 (1.4) U/g protein in the homozygote wild-type, the heterozygote and the homozygote variant type, respectively (p = 0.001). Doctor-diagnosed asthma was unrelated to GPX1 activity in either sex. CONCLUSION: Determinants for activity in the oxidative enzyme GPX1 show marked differences between the sexes, but the activity is not associated with asthma. Sex ought to be taken into consideration when analysing the activity of the enzyme. PMID- 19125370 TI - Active involvement, not illusory control, increases risk taking in a gambling game. AB - The research considers the influence of choice (the possibility for the player to choose a gamble or another) and involvement (the physical interaction with the gambling device) on risk taking in gambling games and whether this influence is mediated by illusory control over the outcome of the gamble. Results of a laboratory experiment (n = 100) show that (a) although choice does increase illusory control, this influence does not translate in increased risk taking, and (b) whilst involvement does increase risk taking, this effect is not mediated by illusory control. These results are discussed in relation to problem gambling, beliefs in the deployability of personal luck, and arousal approaches to risk taking. PMID- 19125371 TI - Mitochondria influence Fas expression in gp120-induced apoptosis of neuronal cells. AB - The human immunodeficiency virus-1 (HIV-1) destroys the immune system and also induces neurological disease culminating into dementia (HIV-associated dementia). Though the HIV viral protein gp120 induces apoptosis in neuronal cells, the mechanism of action is still poorly defined. Recent studies show that cells die during apoptosis by Fas aggregation aided by the mitochondrial proapoptotic proteins. Our studies show an increase in expression of Fas and its associated downstream proteins after treatment of the neuroblastoma cells, SH-SY5Y, with gp120. Fas and its associated death proteins, FADD and caspase-8 (DISC), are downregulated when treated with the caspase inhibitors. The results indicate that mitochondrial-death proteins like caspases may influence the upregulation of the death receptor Fas, and the inhibition of caspases prevents gp120-induced apoptosis. PMID- 19125372 TI - Differential numbers of foci of lymphocytes within the brains of Lewis rats exposed to weak complex nocturnal magnetic fields during development of experimental allergic encephalomyelitis. AB - To discern if specific structures of the rat brain contained more foci of lymphocytes following induction of experimental allergic encephalomyelitis and exposures to weak, amplitude-modulated magnetic fields for 6 min once per hour during the scotophase, the residuals between the observed and predicted values for the numbers of foci for 320 structures were obtained. Compared to the brains of sham-field exposed rats, the brains of rats exposed to 7-Hz 50 nT (0.5 mG) amplitude-modulated fields showed more foci within hippocampal structures and the dorsal central grey of the midbrain while those exposed to 7-Hz 500 nT (5 mG) fields showed greater densities within the hypothalamus and optic chiasm. The brains of rats exposed to either the 50 nT or 500 nT amplitude-modulated 40-Hz fields displayed greater densities of foci within the midbrain structures related to rapid eye movement. Most of the enhancements of infiltrations within the magnetic field-exposed rats occurred in structures within periventricular or periaqueductal regions and were both frequency- and intensity-dependent. The specificity and complexity of the configurations of the residuals of the numbers of infiltrated foci following exposures to the different fields suggest that the brain itself may be a "sensory organ" for the detection of these stimuli. PMID- 19125373 TI - Heat hyperalgesia induced by endoneurial nerve growth factor and the expression of substance P in primary sensory neurons. AB - Endoneurial nerve growth factor (30 ng) produced significant heat hyperalgesia in rats on postinjection days 3 and 5. The percentage of neuron profiles expressing the sensory neuropeptide substance P in the dorsal root ganglion (DRG), and the density and distribution of substance P immunoreactivity at the DRG and the dorsal horn remained essentially unchanged throughout the 10 days of study. NGF increased pain scores in the second phase of the formalin test on postinjection day 3, but not on days 5 and 10. Our results indicate that the observed heat hyperalgesia is not dependent on NGF-induced changes in SP content and release from primary sensory neurons. PMID- 19125374 TI - Effects of progesterone on total brain tissue adenosine deaminase activity in experimental epilepsy. AB - Single seizure and epilepsy is one of the most commonly encountered neurologic disorders in elderly individuals, arising as a result of complex and often multiple acquired underlying pathologies. Adenosine, acting at A1 receptors, exhibits anticonvulsant effects in experimental epilepsy and inhibits progression to status epilepticus. Adenosine deaminase is the enzyme for the regulation of adenosine levels. Therefore any change in adenosine deaminase levels will reflect to adenosine levels. Adenosine deaminase levels were decreased in the groups that were given progesterone. Progesterone may have an antiseizure effect with the additional finding decreased levels of adenosine deaminase that would have resulted in increased adenosine levels that exerts anticonvulsant effect via GABA A receptors. Further studies are needed to evaluate the role of progesterone effects on adenosine deaminase levels and its mechanism(s) in the pathogenesis. PMID- 19125375 TI - Regional difference in stainability with calcium-sensitive acetoxymethyl-ester probes in mouse brain slices. AB - Loading neurons with membrane permeable Ca2+ indicators is a core experimental procedure in functional multineuron Ca2+ imaging (fMCI), an optical technique for monitoring multiple neuronal activities. Although fMCI has been applied to several brain networks, including cerebral cortex, hippocampus, and cerebellum, no studies have systematically addressed the dye-loading efficiency in different brain regions. Here, we describe the stainability of Oregon Green 488 BAPTA-1AM in mouse acute brain slice preparations. The data are suggestive of the potential usability of fMCI in many brain regions, including olfactory bulb, thalamus, dentate gyrus, habenular nucleus, and pons. PMID- 19125376 TI - The levels of glutathione and nitrite-nitrate and the expression of Bcl-2 mRNA in ovariectomized rats treated by raloxifene against kainic acid. AB - The selective estrogen receptor modulators (SERMs) are compounds that activate the estrogen receptors with different estrogenic and antiestrogenic tissue specific effects. The similar effects of SERMs on estrogen encourage the efforts in the research of neuroprotective effects of SERMs. In our study, the potential neuroprotective effects of raloxifene were investigated on the brain cortex of ovariectomized rats after kainic acid-induced oxidative stress. To show the neuroprotective effect of raloxifene against a neurodegenerative agent, kainic acid, expression of Bcl-2, total glutathione (GSH), and nitrite-nitrate levels were investigated in the rat brain cortex. Our results demostrate that raloxifene treatment against oxidative stress significantly increases the expression of Bcl 2 and the level of GSH in the brain cortex. PMID- 19125377 TI - Chaotic patterns of autonomic activity during hypnotic recall. AB - Chaotic neural dynamics likely emerge in cognitive processes and may present time periods that are extremely sensitive to influences affecting the neural system. Recent findings suggest that this sensitivity may increase during retrieval of stressful emotional experiences reflecting underlying mechanism related to consolidation of traumatic memories. In this context, hypnotic recall of anxiety memories in 10 patients, simultaneously with ECG measurement was performed. The same measurement was performed during control cognitive task in 8 anxiety patients and 22 healthy controls. Nonlinear data analysis of ECG records indicates significant increase in the degree of chaos during retrieval of stressful memory in all the patients. The results suggest a role of chaotic neural dynamics during processing of anxiety-related stressful memories. PMID- 19125378 TI - Pien Tze Huang, a composite Chinese traditional herbal extract, affects survival of neuroblastoma cells. AB - Pien Tze Huang is a popular Chinese medicine for liver diseases. In the investigations of possible effects of Pien Tze Huang on the central nervous system, we first studied the in vitro anti-cancer activity of Pien Tze Huang on neuroblastoma cells (SH-SY5Y) as compared with normal fibroblasts (NIH-3T3). Results showed that Pien Tze Huang significantly decreased (p < .05) cell survival of SH-SY5Y as compared to NIH-3T3. Furthermore, the decreases in cell survival of SH-SY5Y were significantly and linearly dose-dependent (p < .05) from 400 to 1,000 microg/ml. This supports further in vivo and animal studies for anti cancer effect, neuroprotection, and their mechanisms. PMID- 19125379 TI - Aromatherapy facts and fictions: a scientific analysis of olfactory effects on mood, physiology and behavior. AB - A systematic review of scientific experimentation addressing olfactory effects on mood, physiology and behavior was undertaken. From this review, 18 studies meeting stringent empirical criteria were then analyzed in detail and it was found that credible evidence that odors can affect mood, physiology and behavior exists. To explain these effects, pharmacological and psychological mechanisms were explored and a psychological interpretation of the data was found to be more comprehensive. Methodological problems regarding dependent measures and stimuli, which led to inconsistencies in the data were discussed, as were the mediating variables of culture, experience, sex differences, and personality. PMID- 19125380 TI - Effects of different levels of food restriction on passive-avoidance memory and the expression of synapsin I in young mice. AB - The present study investigated the effects of food restriction (FR) on memory and the expression of synapsin I in the brain of young mice. The results showed that 20% FR did not retard the body weight gain of mice, while the 60% and 80% FR reduced the mice's body weight. The memory after 24 hr of learning was not changed by FR, whereas long-term memory was improved significantly in 20% FR mice. In addition, 60% and 80% FR did not impair the mice's memory. The transcriptional expression of synapsin I in mice brain was up-regulated by 20% FR, and down-regulated by 60% and 80% FR. PMID- 19125381 TI - Statement of retraction: Synaptic strengthening and continuum activity-wave growth in temporal sequencing during cognitive tasks. PMID- 19125382 TI - Influence of additional cytogenetic abnormalities on the response and survival in late chronic phase chronic myeloid leukemia patients treated with imatinib: long term results. PMID- 19125383 TI - Tumor-specific recombinant idiotype immunisation after chemotherapy as initial treatment for follicular non-Hodgkin lymphoma. AB - Tumor-specific variable regions of the clonal immunoglobulin (idiotype, Id) expressed by B cell non-Hodgkin lymphoma (NHL) can be targeted by active immunotherapy. We conducted a phase I/II trial to determine the safety and immunogenicity of a patient-specific, recombinant, mammalian cell-derived Id protein conjugated to keyhole limpet hemocyanin (Id-KLH; MyVax personalised immunotherapy) in 22 patients with follicular NHL in first remission after chemotherapy. Subjects received five subcutaneous immunisations with MyVax plus locally administered granulocyte-macrophage colony-stimulating factor (GM-CSF). Among 21 evaluable patients, 62% mounted Id-specific immune responses. Evoked anti-Id antibodies recognised both recombinant Id and native Id, and could specifically stain autologous tumor cells. At median follow-up of more than 6 years, median progression-free survival is 38 months. Immunisation of follicular lymphoma patients with MyVax Id-KLH is safe and patients often mount tumor specific immune responses. These results form the basis of a pivotal phase 3 trial of MyVax in follicular NHL. PMID- 19125384 TI - Evaluation of circulating endothelial cells as noninvasive marker of angiogenesis in patients with chronic lymphocytic leukemia. AB - An increased angiogenesis has been documented in bone marrow and lymph nodes of patients with chronic lymphocytic leukemia (CLL). There is accumulating evidence that circulating endothelial cells (CECs) play an important role in angiogenesis. The aim of our study was to compare the number of CECs in peripheral blood of CLL patients and healthy donors and to correlate these numbers with bone marrow microvessel density (MVD) and known prognostic factors in CLL. Proportions of resting CECs (rCECs), activated CECs (aCECs), apoptotic CECs (apoCECs) and circulating precursor endothelial cells (CPECs) were estimated by flow cytometry in 104 untreated CLL patients and 29 healthy blood donors. The MVD was analysed in the bone marrow biopsy in 21 CLL patients and 11 controls using the 'hot spot analysis' of vessels' density (x200 HPS). We found significantly higher numbers of CPECs, rCECs, aCECs and apoCECs in CLL patients than in healthy controls (p < 0.001 for all comparisons). Furthermore, the rCECs number was higher in advanced versus low clinical stage CLL (median 17.5 cells/microL vs. 13.5 cells/microL, p = 0.05). The MVD was significantly higher in the bone marrow of CLL patients when compared with controls (p = 0.016). However, we did not find any correlation between MVD and different CECs populations. In conclusion, the number of CECs is increased in CLL and correlates with stage of the disease, but does not seem to directly reflect the intensity of neovascularisaton in the bone marrow. PMID- 19125385 TI - Early discharge after high-dose melphalan and peripheral blood stem cell reinfusion in patients with hematological and non-hematological disease. AB - The purpose of this study was to analyse our experience of early discharge 2 days after high-dose melphalan (HDM) (Day-1) followed by peripheral blood stem cell re infusion (Day-0) and re-admission on Day +5 in patients with hematological diseases or solid tumors. From 2000 to November 2005, seven patients received tandem Melphalan 200 mg/m(2) HDM with peripheral blood stem cells transplantation (PBSC-T), 130 a single HDM, for a total of 144 procedures. In 123 of them, patients were discharged on Day +1 for re-admission on Day +5 or earlier in the event of complications. Antibiotic prophylaxis was not used. Patients were hospitalised in positive-pressure reverse isolation room during the neutropenic period. Of the 123 procedures eligible for our mixed inpatient-outpatient management regimen, six (5%) required early re-admission for complications. Full engraftment was achieved in all cases. Median time to neutrophil count >0.5 x 10(9)/microL and >1 x 10(9)/microL were 12 and 14 days, respectively. Median time to platelet recovery (>20 x 10(9)/microL) was 13 days. Severe extra-hematological toxicities occurred in 78 (63%) patients: all had oral mucositis and five had associated diarrhoea. During hospitalisation, 94/123 (76%) experienced febrile neutropenia, 20/94 (21%) had documented infection and 74/94 (79%) were considered fever of unknown origin. Median fever duration was 1 day (range 0-11). Median duration of antibiotic treatment was 6 days (range 3-26). Median time to discharge (from Day 0) was 16 days (range 11-57). There was no mortality by on Day +100. Our experience of early discharge after HDM and PBSC-T with re admission on Day +5 is safe and feasible with acceptable frequency of hematological and extra-hematological toxicities. The regimen allows reduced hospital stay and hence cost savings. PMID- 19125386 TI - Current controversies in prenatal diagnosis 4: Should fetal surgery be done in all cases of severe diaphragmatic hernia? PMID- 19125387 TI - Simulating competing risks data in survival analysis. AB - Competing risks analysis considers time-to-first-event ('survival time') and the event type ('cause'), possibly subject to right-censoring. The cause-, i.e. event specific hazards, completely determine the competing risk process, but simulation studies often fall back on the much criticized latent failure time model. Cause specific hazard-driven simulation appears to be the exception; if done, usually only constant hazards are considered, which will be unrealistic in many medical situations. We explain simulating competing risks data based on possibly time dependent cause-specific hazards. The simulation design is as easy as any other, relies on identifiable quantities only and adds to our understanding of the competing risks process. In addition, it immediately generalizes to more complex multistate models. We apply the proposed simulation design to computing the least false parameter of a misspecified proportional subdistribution hazard model, which is a research question of independent interest in competing risks. The simulation specifications have been motivated by data on infectious complications in stem-cell transplanted patients, where results from cause-specific hazards analyses were difficult to interpret in terms of cumulative event probabilities. The simulation illustrates that results from a misspecified proportional subdistribution hazard analysis can be interpreted as a time-averaged effect on the cumulative event probability scale. PMID- 19125388 TI - Properties of the N-terminal domains from Y receptors probed by NMR spectroscopy. AB - Binding of neurohormones from the NPY family to their receptors, the so-called Y receptors, that belong to the superfamily 1b of G-protein coupled receptors might include transient binding to the N-terminal domains of the receptors. Accordingly, we have studied structural features of the N-terminal domains from the Y1, Y2, Y4, and Y5 receptor subtypes (N-Y1, N-Y2, N-Y4, N-Y5). We developed efficient strategies for their recombinant expression. N-Y4 and N-Y1 were expressed as insoluble fusions to enforce accumulation into inclusion bodies, whereas N-Y2 and N-Y5 were expressed as soluble fusion proteins. All N-terminal domains are fully flexible in aqueous buffer. In the presence of phospholipid micelles some stretches within the polypeptides adopt helical conformations, but these are too unstable to be characterized in detail. Using chemical shift mapping techniques, interactions of NPY, peptide YY (PYY), and pancreatic polypeptide (PP), the three members of the neurohormone family that are the Y receptors' natural ligands, with N-Y1, N-Y2, and N-Y5 revealed chemical shift changes in all cases, with the largest values being encountered for PP interacting with N-Y1 or N-Y5 both in the presence and in the absence of phospholipid micelles. The strength of the interactions, however, is generally weak, and the data also point to nonspecific contacts. Previously, in case of the interaction of N-Y4 with PP, the contacts were shown to be electrostatic in nature. This work indicates that association of the peptides with the N-terminal domains may generally be part of their binding trajectory. PMID- 19125389 TI - Noninvasive collection of fresh hairs from free-ranging howler monkeys for DNA extraction. AB - The use of noninvasive collected samples as source of DNA in studies of wild primate populations has increased in recent years. Fresh-plucked hairs represent an important source of DNA, with relatively high quality and concentration. In this study, we describe a low-cost noninvasive technique for collecting fresh plucked hairs used to obtain DNA samples from free-ranging black howler monkey populations (Alouatta pigra). We designed and manufactured darts made of wooden dowels, with the anterior part smeared with glue, which were projected with blowpipes to trap howler monkey hairs. All of the materials to make the darts are inexpensive and are available locally. We collected 89 samples from 76 individuals residing in 15 troops, and the total number of hairs obtained was 754. We found no differences in the number of hairs collected among sex-age classes or among localities but the percentage of darts recovered with sample varied among localities. Preliminary results indicate that over 96% of samples yielded DNA suitable for polymerase chain reaction-based microsatellite marker analysis. The technique proved successful for collecting fresh-plucked hairs of free-ranging black howler monkeys without any trauma to the animals and can be easily adapted to obtain samples from other wild primate and mammal species. PMID- 19125390 TI - Gender moderates the relationship between mania spectrum and serotonin transporter polymorphisms in depression. AB - The short (s) variant of the serotonin transporter gene linked functional polymorphic region (5-HTTLPR) is associated with depression. Stressful life events, gender, and race have been shown to moderate this association. Because features of mania/hypomania seem to constitute an indicator of higher severity of depression, we examined the relationship between 5-HTTLPR genotype and symptoms of mania-hypomania spectrum occurring over the lifetime in patients with major depression. The possible moderating role of gender in this relationship was taken into account. Two hundred twenty-two patients with unipolar major depression were genotyped for 5-HTTLPR and nine other representative polymorphisms, and were administered the Mood Spectrum Questionnaire, Lifetime Version (MOODS-SR). The manic-hypomanic (MH) component score was used for analysis. Using a linear model of the MH score as a function of genotypes and gender, controlling for age, severity of depression, and site, we found significant effects of gender (F = 8.003, df = 1, P = 0.005), of the interaction gender x genotype (F = 4.505, df = 2, P = 0.012), and of the baseline Hamilton score (F = 5.404, df = 1, P = 0.021), non-significant effects of genotype (F = 1.298, df = 2, P = 0.275), age (F = 0.310, df = 1, P = 0.578) site (F = 0.504, df = 1, P = 0.479). Significant associations were also detected at three other SNPs. The association between the manic/hypomanic component of the MOODS-SR and the polymorphisms of the 5-HTTLPR is moderated by gender. This finding is intriguing from a clinical point of view because women with unipolar disorder and the "ss" genotype seem to constitute a sub-group with higher severity of depression. These results should be considered tentative pending replication in other samples. PMID- 19125392 TI - Measures of agreement for vectorcardiography data. AB - In this paper, nonparametric methods are proposed for quantifying agreement and disagreement between different measurement methods when the results of the measurements are rotation matrices. First, the expected squared distance between two matrices is used to quantify the measurement agreement. Two choices of such distance are considered-the Frobenius distance and geodesic distance. Second, the notion of 'concordance correlation coefficient', a commonly used measure of agreement, is extended to the space of rotation matrices. Such generalized concordance coefficient can be treated as a normalized expected squared distance. Since no two measurement systems can be expected to be in perfect agreement, it becomes necessary to define a notion of practical agreement. We define such a notion. Moreover, for both proposed methods, the percentile bootstrap procedure is implemented to provide a confidence interval to help make a decision concerning practical agreement/disagreement in real-life applications. The methodology is illustrated using two data sets, one based on an application involving vectorcardiography data (Biometrika 1972; 59:665-676) and the other based on a synthetic data set. PMID- 19125391 TI - Apparent diffusion coefficient and fractional anisotropy of newly diagnosed grade II gliomas. AB - Distinguishing between low-grade oligodendrogliomas (ODs) and astrocytomas (AC) is of interest for defining prognosis and stratifying patients to specific treatment regimens. The purpose of this study was to determine if the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) from diffusion imaging can help to differentiate between newly diagnosed grade II OD and AC subtypes and to evaluate the ADC and FA values for the mixed population of oligoastrocytomas (OA). Fifty-three patients with newly diagnosed grade II gliomas were studied using a 1.5T whole body scanner (23 ODs, 16 ACs, and 14 OAs). The imaging protocol included post-gadolinium T1-weighted images, T2-weighted images, and either three and/or six directional diffusion imaging sequence with b = 1000 s/mm(2). Diffusion-weighted images were analyzed using in-house software to calculate maps of ADC and for six directional acquisitions, FA. The intensity values were normalized by values from normal appearing white matter (NAWM) to generate maps of normalized apparent diffusion coefficient (nADC) and normalized fractional anisotropy (nFA). The hyperintense region in the T2 weighted image was defined as the T2All region. A Mann-Whitney rank-sum test was performed on the 25th, median, and 75th nADC and nFA among the three subtypes. Logistic regression was performed to determine how well the nADC and nFA predict subtype. Lesions diagnosed as being OD had significantly lower nADC and significantly higher nFA, compared to AC. The nADC and nFA values individually classified the data with an accuracy of 87%. Combining the two did not enhance the classification. The patients with OA had nADC and nFA values between those of OD and AC. This suggests that ADC and FA may be helpful in directing tissue sampling to the most appropriate regions for taking biopsies in order to make a definitive diagnosis. PMID- 19125393 TI - Comments on 'Rebuttal to Carpenter et al.' Comments on 'Fixed vs random effects meta-analysis in rare event studies: the rosiglitazone link with myocardial infarction and cardiac death' by J. J. Shuster, L. S. Jones and D. A. Salmon, Statistics in Medicine 2008; 27:3912-3914. PMID- 19125394 TI - Comments on 'On estimating treatment effects under non-compliance in randomized clinical trials: are intent-to-treat or instrumental variables analyses perfect solutions?' by Heejung Bang and Clarence E. Davis, Statistics in Medicine 2007; 26:954-964. PMID- 19125396 TI - Comments on 'Tests for the homogeneity of two binomial proportions in extremely unbalanced 2x2 contingency tables', by S.-H. Kang and C. W. Ahn, Statistics in Medicine 2008; 27:2524-2535. PMID- 19125397 TI - Reactivity of Tyr-Leu and Leu-Tyr dipeptides: identification of oxidation products by liquid chromatography-tandem mass spectrometry. AB - The exposure of peptides and proteins to reactive hydroxyl radicals results in covalent modifications of amino acid side-chains and protein backbone. In this study we have investigated the oxidation the isomeric peptides tyrosine-leucine (YL) and leucine-tyrosine (LY), by the hydroxyl radical formed under Fenton reaction (Fe(2+)/H(2)O(2)). Through mass spectrometry (MS), high-performance liquid chromatography (HPLC-MS) and electrospray tandem mass spectrometry (HPLC MS(n)) measurements, we have identified and characterized the oxidation products of these two dipeptides. This approach allowed observing and identifying a wide variety of oxidation products, including isomeric forms of the oxidized dipeptides. We detected oxidation products with 1, 2, 3 and 4 oxygen atoms for both peptides; however, oxidation products with 5 oxygen atoms were only present in LY. LY dipeptide oxidation leads to more isomers with 1 and 2 oxygen atoms than YL (3 vs 5 and 4 vs 5, respectively). Formation of the peroxy group occurred preferentially in the C-terminal residue. We have also detected oxidation products with double bonds or keto groups, dimers (YL-YL and LY-LY) and other products as a result of cross-linking. Both amino acids in the dipeptides were oxidized although the peptides showed different oxidation products. Also, amino acid residues have shown different oxidation products depending on the relative position on the dipeptide. Results suggest that amino acids in the C-terminal position are more prone to oxidation. PMID- 19125399 TI - Metabolites from the biodegradation of 1,6-hexanediol dibenzoate, a potential green plasticizer, by Rhodococcus rhodochrous. AB - Metabolites from the biodegradation of a potential plasticizer 1,6-hexanediol dibenzoate in the presence of n-hexadecane as a co-substrate by the common soil organism Rhodococcus rhodochrous were identified using GC/MS and Fourier transform mass spectroscopy (FTMS) techniques. Trimethylsilylation of compounds from the biodegradation broth permitted detection of the following metabolites: 1 hexadecyl benzoate, 6-benzoyloxyhexanoic acid, 4-benzoyloxybutanoic acid, 6 benzoyloxyhexan-1-ol and benzoic acid. The presence of these metabolites was confirmed by repeating the biodegradation with 1,6-hexanediol di[(2)H(5)]benzoate, by measurement of their exact masses in FTMS and by comparison with available authentic materials. The results show that biodegradation of 1,6-hexanediol dibenzoate by R. rhodochrous does not lead to the accumulation of persistent metabolites as has been reported for commercial dibenzoate plasticizers. PMID- 19125400 TI - Dissociative photoionization of isoprene: experiments and calculations. AB - Vacuum ultraviolet (VUV) dissociative photoionization of isoprene in the energy region 8.5-18 eV was investigated with photoionization mass spectroscopy (PIMS) using synchrotron radiation (SR). The ionization energy (IE) of isoprene as well as the appearance energies (AEs) of its fragment ions C(5)H(7) (+), C(5)H(5) (+), C(4)H(5) (+), C(3)H(6) (+), C(3)H(5) (+), C(3)H(4) (+), C(3)H(3) (+) and C(2)H(3) (+) were determined with photoionization efficiency (PIE) curves. The dissociation energies of some possible dissociation channels to produce those fragment ions were also determined experimentally. The total energies of C(5)H(8) and its main fragments were calculated using the Gaussian 03 program and the Gaussian-2 method. The IE of C(5)H(8), the AEs for its fragment ions, and the dissociation energies to produce them were predicted using the high-accuracy energy model. According to our results, the experimental dissociation energies were in reasonable agreement with the calculated values of the proposed photodissociation channels of C(5)H(8). PMID- 19125398 TI - Identification of intact oxidation products of glycerophospholipids in vitro and in vivo using negative ion electrospray iontrap mass spectrometry. AB - Free radical-induced oxidation products of polyunsaturated fatty acids esterified to phospholipids have been implicated in a number of human diseases including atherosclerosis and neurodegenerative diseases. Some of these phospholipid oxidation products have potent biological activities and likely contribute to human pathophysiological conditions. Oxidation products have also been used as markers of oxidative stress in vivo. Identification and quantification of phospholipid oxidation products are often performed by analyzing the oxidized free fatty acid moieties after hydrolysis from the phospholipids head groups by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS). We now describe the definitive identification of intact oxidized products of glycerophospholipids including glycerophosphatidylcholine (GPC), glycerophosphatidylethanolamine (GPE), and glycerophosphatidylserine (GPS) in vitro and in vivo using iontrap MS. For these analyses, the negative ions of the oxidation products of phospholipids are fragmented to MS(n) and unequivocal structural characterization is obtained based on collision-induced dissociation (CID) of the sn-2 carboxylate ion. This technique overcomes the need to hydrolyze fatty acids from phospholipids in the analysis. The method has been used to identify a number of oxidation products of glycerophospholipids including hydroxyeicosatetraenoates (HETEs) and isoprostanes (IsoPs) esterified to different classes of glycerophospholipids in vitro and in vivo. These studies thus provide a new approach to identify the intact oxidation products of glycerolphospholipids. PMID- 19125401 TI - A systematic review of research examining benzodiazepine-related mortality. AB - PURPOSE: This paper will review literature examining the association of benzodiazepine use and mortality. METHODS: An extensive literature review was undertaken to locate all English-language published articles that examine mortality risk associated with use of benzodiazepines from 1990 onwards. RESULTS: Six cohort studies meeting the criteria above were identified. The results were mixed. Three of the studies assessed elderly populations and did not find an increased risk of death associated with benzodiazepine use, whereas another study of the general population did find an increased risk, particularly for older age groups. A study of a middle aged population found that regular benzodiazepine use was associated with an increased mortality risk, and a study of 'drug misusers' found a significant relationship between regular use of non-prescribed benzodiazepines and fatal overdose. Three retrospective population-based registry studies were also identified. The first unveiled a high relative risk (RR) of death due to benzodiazepine poisoning versus other outcomes in patients 60 or older when compared to those under 60. A positive but non-significant association between benzodiazepine use and driver-responsible fatalities in on-road motor vehicle accidents was reported. Drug poisoning deaths in England showed benzodiazepines caused 3.8% of all deaths caused by poisoning from a single drug. CONCLUSION: On the basis of existing research there is limited data examining independent effects of illicit benzodiazepine use upon mortality. Future research is needed to carefully examine risks of use in accordance with doctors' prescriptions and extra-medical use. PMID- 19125402 TI - Metal contamination in matrix-assisted laser desorption/ionization samples prepared with the 'vortex' solvent-free method. PMID- 19125403 TI - On the use of hydrogen/deuterium exchange mass spectrometry data to improve de novo protein structure prediction. PMID- 19125404 TI - Expression and molecular diversity of Tcf7l2 in the developing murine cerebellum and brain. AB - The Wingless family of secreted proteins impinges on multiple aspects of vertebrate nervous system development, from early global patterning and cell fate decision to synaptogenesis. Here, we mapped the developmental expression of the Tcf7l2, which is key to the canonical Wingless signaling cascade, in the developing cerebellum. The exclusive and transient expression of Tcf7l2 in ventricular and Olig2-defined precursor cells within the cerebellar anlage, and its predominant expression in postmitotic neurons in the midbrain/inferior colliculus allowed us to ask whether cell type-specific differences are also reflected in splice isoform variability. We also included in this analysis intestinal epithelia, where Tcf7l2 function has been intensively studied. Our data reveal extensive variability of Tcf7l2 splicing in the central nervous system. Additional variability in brain-expressed Tcf7l2 is generated by a length polymorphism of expressed mRNAs in a stretch of normally nine adenines found at the beginning of exon 18, reminiscent of variability observed at the same site in cancers with microsatellite instability. A consensus emerging from our data is that the expression of isoforms comprising or lacking the C-clamp motif, which has been linked by in vitro studies to the regulation of cell growth, is indeed tightly correlated with the proliferative status in vivo. PMID- 19125405 TI - BCL-2 and BAX proteins expression throughout the light-dark cycle and modifications induced by sleep deprivation and rebound in adult rat brain. AB - It has been suggested that sleep has a restorative function; however, experimental support is limited. Hence, we investigated whether changes in the level of antiapoptotic BCL-2 protein and proapoptotic BAX protein occur during sleep deprivation (SD) and sleep rebound, and evaluated the spontaneous changes in these proteins, along the light-dark cycle, in the adult male Wistar rat. Estimations were made in the prefrontal cortex, hippocampus, striatum, and pons. We observed that BCL-2 exhibited diurnal variations in the prefrontal cortex and striatum, whereas BAX varied in the striatum and showed only small variations in the pons as measured by immunoblotting. The BCL-2/BAX ratio exhibited diurnal variations in the prefrontal cortex and striatum. BCL-2 and BAX levels were affected by 24 hr of total SD and 24 hr of sleep rebound. SD decreased the BCL 2/BAX ratio in the prefrontal cortex and pons. Sleep rebound increased the BCL 2/BAX ratio in the hippocampus. In conclusion, the BCL-2/BAX ratio is high during the dark phase as compared with the light phase in the prefrontal cortex and during the light phase as compared with the dark phase in the striatum. SD decreased the BCL-2/BAX ratio in the prefrontal cortex and pons, whereas sleep rebound increased it in the hippocampus. These changes point out structures in the brain that express these proteins as a response to the light-dark cycle. Similarly, SD and sleep rebound seem to change these proteins expression in some other brain structures, suggesting that cellular vulnerability might be altered by these changes. PMID- 19125406 TI - The myelinated fiber changes in the white matter of aged female Long-Evans rats. AB - Age-related changes of the white matter and the myelinated fibers in white matter of female Long-Evans rats were investigated by the use of design-based stereological methods. The white matter volume in middle-aged rats was not significantly different from that in young rats, but the white matter volume in aged rats was significantly decreased by 42.7% and 37.7%, respectively, when compared with young and middle-aged rats. When compared with young rats, the total volumes of the myelinated fibers and myelin sheaths in white matter of aged rats were significantly decreased by 26.7% and 30.7%, respectively. When compared with young and middle-aged rats, the total length of the myelinated fibers in white matter of aged rats was significantly decreased by 52.5% and 48.6%, respectively. The mean diameter of the myelinated fibers in white matter of aged rats was significantly increased by 37.5% when compared with young rats. The absolute diameter distributions of the myelinated fiber length in the white matter of young, middle-aged, and aged rats indicated that the age-related loss of the myelinated fiber length in the white matter of aged rats was mainly as a result of the marked loss of the myelinated fibers with smaller diameter. The age related changes of the white matter and the myelinated fibers in white matter may have important implications for age-related cognitive impairments. PMID- 19125407 TI - A novel method for isolating Schwann cells using the extracellular domain of Necl1. AB - Myelinating cocultures of Schwann cells and dorsal root ganglion neurons are a powerful experimental system for probing the molecular mechanisms of axon-Schwann cell interaction. The isolation of a pure population of myelination-competent Schwann cells is a prerequisite for this experimental system. We describe here a protocol for a FACS-based isolation of Schwann cells utilizing a specific affinity reagent (Necl1-Fc) and the use of these isolated cells in myelinating cocultures. An advantage of the myelinating coculture system is that Schwann cells and the neurons can be genetically manipulated before they are cocultured. We further show that our method allows the isolation of virally transduced Schwann cells in a single purification step. This protocol for the FACS-based isolation of myelination-competent Schwann cells by Necl1-Fc and the use of these cells in myelinating cocultures should significantly facilitate future studies aimed at delineation of the molecular mechanisms of axon-Schwann cell interactions and myelination. PMID- 19125410 TI - Reciprocal regulation of ATPgammaS-induced monocyte chemoattractant protein-1 production by ERK and p38 MAP kinases in rat corticostriatal slice cultures. AB - Monocyte chemoattractant protein-1 (MCP-1, CCL2) is a well-defined chemokine implicated in the pathology of various neurodegenerative diseases and brain injuries, such as Alzheimer's disease, multiple sclerosis, stroke, and traumatic injury. We investigated the effect of the activation of P2 purinoceptors on MCP-1 production in rat corticostriatal slice cultures. Treatment with adenosine 5'-O (3-thiotriphosphate) (ATPgammaS), a hydrolysis-resistant adenosine triphosphate (ATP) analog, induced MCP-1 production in astrocytes. The induction was in a concentration-dependent manner and was antagonized by a P2 purinoceptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid. The inhibition of an extracellular signal-regulated kinase (ERK) pathway by PD98059 and U0126 significantly suppressed ATPgammaS-induced MCP-1 mRNA expression and protein production, while inhibition of c-Jun N-terminal kinase by SP600125 resulted in the partial suppression. Conversely, SB203580, a p38 mitogen-activated protein (MAP) kinase inhibitor, significantly enhanced ATPgammaS-induced MCP-1 production. Similar effects of ERK and p38 MAP kinase inhibitors on MCP-1 production were observed in the slices stimulated by ATP and BzATP. These results demonstrate that astrocytic MCP-1 production induced by P2 purinoceptor stimulation is reciprocally regulated by ERK and p38 MAP kinases in the organotypic slice cultures. PMID- 19125408 TI - Extensive spinal decussation and bilateral termination of cervical corticospinal projections in rhesus monkeys. AB - To examine neuroanatomical mechanisms underlying fine motor control of the primate hand, adult rhesus monkeys underwent injections of biotinylated dextran amine (BDA) into the right motor cortex. Spinal axonal anatomy was examined using detailed serial-section reconstruction and modified stereological quantification. Eighty-seven percent of corticospinal tract (CST) axons decussated in the medullary pyramids and descended through the contralateral dorsolateral tract of the spinal cord. Eleven percent of CST axons projected through the dorsolateral CST ipsilateral to the hemisphere of origin, and 2% of axons projected through the ipsilateral ventromedial CST. Notably, corticospinal axons decussated extensively across the spinal cord midline. Remarkably, nearly 2-fold more CST axons decussated across the cervical spinal cord midline (approximately 12,000 axons) than were labeled in all descending components of the CST (approximately 6,700 axons). These findings suggest that CST axons extend multiple segmental collaterals. Furthermore, serial-section reconstructions revealed that individual axons descending in either the ipsilateral or contralateral dorsolateral CST can: 1) terminate in the gray matter ipsilateral to the hemisphere of origin; 2) terminate in the gray matter contralateral to the hemisphere of origin; or 3) branch in the spinal cord and terminate on both sides of the spinal cord. These results reveal a previously unappreciated degree of bilaterality and complexity of corticospinal projections in the primate spinal cord. This bilaterality is more extensive than that of the rat CST, and may resemble human CST organization. Thus, augmentation of sprouting of these extensive bilateral CST projections may provide a novel target for enhancing recovery after spinal cord injury. PMID- 19125409 TI - Role of monocyte chemoattractant protein-1 (MCP-1/CCL2) in migration of neural progenitor cells toward glial tumors. AB - Neural progenitor cells (NPCs) have been investigated as potential vehicles for brain tumor therapy because they have been shown to migrate toward central nervous system gliomas and can be genetically engineered to deliver cytotoxic agents to tumors. The mechanisms that regulate migration of NPCs to tumors are not fully understood. By means of microarray analysis, polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry, we found that monocyte chemoattractant protein-1 (MCP-1/CCL-2) was expressed in experimental brain tumor cells in vivo and in vitro. CCR2, the receptor for MCP-1, was expressed on C17.2 NPCs. We used a modified Boyden chamber assay and found increased migration of NPCs in vitro in response to MCP-1. By means of an in vivo model for NPC migration, we found evidence of NPC migration toward areas of MCP-1 infusion in rat brains. An understanding of NPC migration mechanisms may be used to enhance delivery of cytotoxic agents to brain tumor cells. PMID- 19125411 TI - Depletion of CD4(+)CD25(+) T cells exacerbates experimental autoimmune encephalomyelitis induced by mouse, but not rat, antigens. AB - A key question in the field of autoimmunity concerns the fact that experimental disease is generally induced more easily with closely related, but not completely identical, tissue-restricted antigens. Here, the possibility that naturally occurring regulatory T cells (Tregs) for self-antigens are more potent than those for related antigens was investigated. The self-antigen specificity of naturally occurring Tregs was tested in experimental autoimmune encephalomyelitis (EAE) induced with mouse (self) or closely related (rat) myelin oligodendrocyte glycoproteins (MOGs). Surprisingly, Treg depletion increased EAE severity in mice immunized with mouse, but not rat, MOG. This increase was associated with increased T-cell activation and infiltration of the central nervous system, as well as increased interleukin (IL)-17 production and a higher ratio of interferon gamma- to IL-10-producing cells. These data suggest that Tregs are specific for self-antigen and do not "cross-protect" against autoimmunity even when disease is induced with closely related foreign antigens. PMID- 19125413 TI - Analysis of oligomeric peroxides in synthetic triacetone triperoxide samples by tandem mass spectrometry. AB - Oligomeric peroxides formed in the synthesis of triacetone triperoxide (TATP) have been analyzed by mass spectrometry utilizing both electrospray ionization (ESI) and chemical ionization (CI) to form sodiated adducts (by ESI) and ammonium adducts (by CI and ESI). Tandem mass spectrometry and deuterium isotopic labeling experiments have been used to elucidate the collision-induced dissociation (CID) mechanisms for the adducts. The CID mechanisms differ for the sodium and ammonium adducts and vary with the size of the oligoperoxide. The sodium adducts of the oligoperoxides, H[OOC(CH(3))(2)](n)OOH, do not cyclize under CID, whereas the ammonium adducts of the smaller oligoperoides (n < 6) do form the cyclic peroxides under CID. Larger oligoperoxide adducts with both sodium and ammonium undergo dissociation through cleavage of the backbone under CID to form acyl- and hydroperoxy-terminated oligomers of the general form CH(3)C(O)[OOC(CH(3))(2)](x)OOH, where x is an integer less than the original oligoperoxide degree of oligomerization. The oligoperoxide distribution is shown to vary batch-to-batch in the synthesis of TATP and the post-blast distribution differs slightly from the distribution in the uninitiated material. The oligoperoxides are shown to be decomposed under gentle heating. PMID- 19125412 TI - Expression of estrogen receptor GPR30 in the rat spinal cord and in autonomic and sensory ganglia. AB - The G protein-coupled receptor GPR30 has recently been identified as a nonnuclear estrogen receptor. Reverse transcriptase-polymerase chain reaction revealed expression of GPR30 mRNA in varying quantities in the rat spinal cord, dorsal root ganglia, nodose ganglia, trigeminal ganglia, hippocampus, brain stem, and hypothalamus. Immunohistochemical studies that used a rabbit polyclonal antiserum against the human GPR30 C-terminus revealed a fine network of GPR30 immunoreactive (irGPR30) cell processes in the superficial layers of the spinal cord; some of which extended into deeper laminae. A population of neurons in the dorsal horn and ventral horn were irGPR30. Dorsal root, nodose, and trigeminal ganglionic neurons displayed varying intensities of irGPR30. Positively labeled neurons were detected in the major pelvic ganglion, but not in the superior cervical ganglion. A population of chromaffin cells in the adrenal medulla was irGPR30, so were cells of the zona glomerulosa. Double-labeling the adrenal medulla with GPR30 antiserum and tyrosine hydroxylase antibody or phenylethanolamine-N-methyltransferase antiserum revealed that irGPR30 is expressed in the majority of tyrosine hydroxylase-positive chromaffin cells. Last, some of the myenteric ganglion cells were irGPR30. Tissues processed with preimmune serum resulted in no staining. Voltage-sensitive dye imaging studies showed that the selective GPR30 agonist G-1 (1, 10, and 100 nM) depolarized cultured spinal neurons in a concentration-dependent manner. Collectively, our result provides the first evidence that GPR30 is expressed in neurons of the dorsal and ventral horn as well as in sensory and autonomic neurons, and activation of GPR30 by the selective agonist G-1 depolarizes cultured spinal neurons. PMID- 19125414 TI - The detection of various opiates and benzodiazepines by comprehensive two dimensional gas chromatography/time-of-flight mass spectrometry. AB - A technique using comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC x GC/TOFMS) is applied to qualitative and quantitative drug testing. Human serum was 'spiked' with known quantities of benzodiazepines and a 'street heroin' mixture including some of the major metabolites and impurities. The sample components were extracted from the matrix by solid-phase extraction (SPE). Constituents containing polar hydroxyl and/or secondary amine groups were derivatised with N-methyl-N-(tert-butyldimethyl)trifluoroacetamide (MTBSTFA) to improve the chromatographic performance. An orthogonal separation of the matrix constituents was achieved by coupling a DB-5ms (5% phenyl) to a BPX50 (50% phenyl) GC column. The eluant was focused onto the second column by a twin-stage cryo-modulator. Rapid 6 s modulation times were achieved by transfer from a 30 m x 0.25 mm (length x internal diameter) to a 2 m x 0.1 mm column. TOFMS with rapid spectral acquisition (< or =500 spectra/s) was employed in the mass range m/z 40 650. A clean mass spectrum was obtained for each analyte using mass spectral deconvolution software. The sensitivity and repeatability of the method were evaluated by the preparation of calibration standards for two benzodiazepines, flunitrazepam and its major metabolite 7-aminoflunitrazepam (7-amino-FN), in the concentration range 5-1000 ng/mL. The limits of detection (LODs) and limits of quantitation (LOQs), calculated by repeat injections (x10) of the lowest standard, were 1.6 and 5.4 ng/mL (flunitrazepam); 2.5 and 8.5 ng/mL (7-amino-FN), respectively. There is scope to extend this protocol to screen a large number of drugs and metabolites stored in a library database. PMID- 19125415 TI - Low magnitude and high frequency mechanical loading prevents decreased bone formation responses of 2T3 preosteoblasts. AB - Bone loss due to osteoporosis or disuse such as in paraplegia or microgravity is a significant health problem. As a treatment for osteoporosis, brief exposure of intact animals or humans to low magnitude and high frequency (LMHF) mechanical loading has been shown to normalize and prevent bone loss. However, the underlying molecular changes and the target cells by which LMHF mechanical loading alleviate bone loss are not known. Here, we hypothesized that direct application of LMHF mechanical loading to osteoblasts alters their cell responses, preventing decreased bone formation induced by disuse or microgravity conditions. To test our hypothesis, preosteoblast 2T3 cells were exposed to a disuse condition using the random positioning machine (RPM) and intervened with an LMHF mechanical load (0.1-0.4 g at 30 Hz for 10-60 min/day). Exposure of 2T3 cells to the RPM decreased bone formation responses as determined by alkaline phosphatase (ALP) activity and mineralization even in the presence of a submaximal dose of BMP4 (20 ng/ml). However, LMHF mechanical loading prevented the RPM-induced decrease in ALP activity and mineralization. Mineralization induced by LMHF mechanical loading was enhanced by treatment with bone morphogenic protein 4 (BMP4) and blocked by the BMP antagonist noggin, suggesting a role for BMPs in this response. In addition, LMHF mechanical loading rescued the RPM-induced decrease in gene expression of ALP, runx2, osteomodulin, parathyroid hormone receptor 1, and osteoglycin. These findings suggest that preosteoblasts may directly respond to LMHF mechanical loading to induce differentiation responses. The mechanosensitive genes identified here provide potential targets for pharmaceutical treatments that may be used in combination with low level mechanical loading to better treat osteoporosis or disuse-induced bone loss. PMID- 19125416 TI - Nrf2 activators provide neuroprotection against 6-hydroxydopamine toxicity in rat organotypic nigrostriatal cocultures. AB - Oxidative stress and inflammation appear to play a critical role in the progression of Parkinson's disease. As a result, there has been growing interest in antioxidant pathways and how these pathways might be exploited to slow the progressive loss of dopamine neurons. One such pathway that has garnered attention recently is mediated by the transcription factor Nrf2 and is integral in orchestrating cells' antiinflammatory defense. Nrf2 controls the inducible expression of numerous antioxidant and phase 2 detoxification genes, such as glutathione S-transferase, heme oxygenase-1, and NAD(P)H:quinone oxidoreductase 1 (NQO1). Once activated, these genes work synergistically to maintain intracellular redox homeostasis. In this study, we test the hypothesis that Nrf2 activation can protect dopaminergic neurons against 6-hydroxydopamine (6-OHDA) induced toxicity. Treatment of organotypic nigrostriatal cocultures with either tert-butylhydroquinone (tBHQ) or sulforaphane, known activators of Nrf2, mitigated dopaminergic cell loss. The observed protection appeared to be mediated, at least in part, by an increase in antioxidant activity. Simultaneous treatment of cultures with tBHQ and 6-OHDA increased NQO1 expression 17-fold compared with controls. Overall, these results suggest that Nrf2 may play an important role in cellular protection in neurodegenerative diseases and may be a viable therapeutic target in the future. PMID- 19125417 TI - Shared TP53 gene mutation in morphologically and phenotypically distinct concurrent primary small cell neuroendocrine carcinoma and adenocarcinoma of the prostate. AB - BACKGROUND: Small cell carcinoma of the prostate is an uncommon neoplasm, the origin of which has been controversial. To address this, we performed transcriptome profiling and TP53 sequencing of concurrent small cell and prostatic adenocarcinoma to determine the relationship between these entities. METHODS: We identified an unusual case of primary prostate cancer that contained adjacent acinar adenocarcinoma (Gleason score 4 + 3 = 7) and small cell carcinoma. We performed laser capture microdissection to isolate tumor components and performed gene expression and TP53 gene sequence analysis on each component, with results validated by immunohistochemistry for PSA, PSAP, PSMA, androgen receptor, NKX 3.1 and neuroendocrine markers. RESULTS: Transcriptome profiling of the carcinoma components identified 99 genes with a greater than 10-fold differential expression between prostatic adenocarcinoma and small cell carcinoma, many of which have not been previously reported in prostate cancer. The small cell carcinoma component demonstrated upregulation of proliferative and neuroendocrine markers and tyrosine kinase receptors, and downregulation of cell adhesion molecules, supporting the aggressive nature of this form of carcinoma. Sequencing of the TP53 gene suggested a common clonal origin for both components. CONCLUSIONS: This is the first report of a primary small cell carcinoma of the prostate subjected to extensive molecular analysis and the first to show a clonal relation between two morphologically distinct prostate cancer types. The evidence of progression to small cell carcinoma may yield important insights into the pathogenesis of this entity and provide a novel spectrum of molecular markers to further dissect cellular pathways important in tumor progression. PMID- 19125419 TI - KCTD10 interacts with proliferating cell nuclear antigen and its down-regulation could inhibit cell proliferation. AB - A novel gene (GenBank accession No. AF113208) named KCTD10 (potassium channel tetramerisation domain-containing 10) was cloned from our 5300 EST database of human aorta cDNA library. Computational analysis showed that KCTD10 cDNA is 2,638 bp long, encoding 313 amino acids with a proliferating cell nuclear antigen binding motif, mapped to chromosome 12q24.11 with 7 exons, ubiquitously expressed in all 12 tested normal tissues and 7 of 8 tested tumor cell lines from MTN membranes by Northern blot. Nuclear localization of KCTD10 was observed in A549 cells. Yeast two-hybrid analysis and immunoprecipitation assay showed that KCTD10 can interact with PCNA. In A549 cells, KCTD10 down-regulation could inhibit cell proliferation, but its over-expression could not influence cell proliferation. The results suggest that KCTD10 may be associated with DNA synthesis and cell proliferation. PMID- 19125418 TI - AMP-activated protein kinase suppresses LXR-dependent sterol regulatory element binding protein-1c transcription in rat hepatoma McA-RH7777 cells. AB - AMP-activated protein kinase (AMPK) is an intracellular fuel sensor that plays a key role in regulating fatty acid synthesis in liver. Sterol regulatory element binding protein (SREBP)-1c is a master regulator of hepatic lipogenic gene expression. It has long been documented that AMPK activation suppresses hepatic SREBP-1 mRNA and nuclear SREBP-1 protein. But the mechanism remains undefined. In this study we investigated the molecular mechanisms by which AMPK downregulates hepatic SREBP-1c mRNA using a novel model cell line McA-RH7777. We found that AMPK is robustly activated in rat hepatoma McA-RH7777 cells treated with two widely used AMPK activators, AICAR and metformin, and AMPK activation sharply suppresses SREBP-1c mRNA and nuclear SREBP-1c protein, but not SREBP-1a mRNA derived from the same gene. These inhibitory effects are reversed by the AMPK inhibitor Compound C or 8-BrAMP, demonstrating the requirement of AMPK in the suppression of SREBP-1c mRNA and nuclear SREBP-1c protein by AICAR and metformin. AMPK does not enhance SREBP-1c mRNA degradation in the presence of the general transcription inhibitor actinomycin D; instead it inhibits SREBP-1c promoter activity in a luciferase reporter assay. AMPK-mediated inhibition of SREBP-1c promoter activity can also be abrogated by the AMPK inhibitor Compound C. Furthermore AMPK activation significantly attenuates the synthetic liver X receptor (LXR) ligand T0901317-induced SREBP-1c promoter activity. AMPK also inhibits cleavage of LXR ligand-induced SREBP-1c precursor. We conclude that AMPK suppresses hepatic SREBP-1c mRNA expression by inhibiting LXR-dependent SREBP-1c transcription via inhibition of endogenous LXR ligand production and by inhibiting SREBP-1c processing in McA-RH7777 cells. PMID- 19125420 TI - Rapid detection of drugs in biofluids using atmospheric pressure chemi/chemical ionization mass spectrometry. AB - We have demonstrated that, with simple pH adjustment, volatile drugs such as methamphetamine, amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), ketamine, and valproic acid could be analyzed rapidly from raw biofluid samples (e.g. urine and serum) without dilution, or extraction, using atmospheric pressure ionization. The ion source was a variant type of atmospheric pressure chemical ionization (APCI) that used a dielectric barrier discharge (DBD) to generate the metastable helium gas and reagent ions. The sample solution was loaded in a disposable glass pipette, and the volatile compounds were purged by nitrogen gas to be reacted with the metastable helium gas. The electrodes of the DBD were arranged in such a way that the generated glow discharge was confined within the discharge tube and was not exposed to the analytes. A needle held at 100-500 V was placed between the ion-sampling orifice and the discharge tube to guide the analyte ions into the mass spectrometer. After pH adjustment of the biofluid sample, the amphiphilic drugs were in the form of a water-insoluble oil, which could be concentrated on the liquid surface. By gentle heating of the sample to increase the evaporation rate, rapid and sensitive detection of these drugs in raw urine and serum samples could be achieved in less than 2 min for each sample. PMID- 19125422 TI - Overexpression of 5 alpha-reductase type 1 increases sensitivity of prostate cancer cells to low concentrations of testosterone. AB - INTRODUCTION: Conversion of testosterone to dihydrotestosterone (DHT) by the enzymes 5 alpha-reductase types 1 (5 alpha R1) and 2 (5 alpha R2) is important for normal and pathological growth of the prostate. The predominant isoenzyme in normal prostate is 5 alpha R2. However, prostate cancer (PCa) development is accompanied by a decrease in 5 alpha R2 and an increase in 5 alpha R1. The biological significance of increased 5 alpha R1 expression is not fully understood. Therefore, the aim of this study was to determine the effect of overexpression of 5 alpha R1 on growth and prostate-specific antigen (PSA) production in PCa cells. MATERIALS AND METHODS: LNGK-9 PCa cells, transiently transfected with pTRE-5 alpha R1 or pTRE alone, were cultured in the presence or absence of testosterone at varying concentrations. Cell growth and PSA secretion were measured after 4-6 days. Cyclin E1, Ki67, and PSA mRNA levels were evaluated using RT-PCR after 24 hr of treatment. RESULTS: 10 pM testosterone increased growth of pTRE-5 alpha R1 transfectants by 54.1% over cells grown in the absence of testosterone, compared to 25.0% in pTRE transfectants (P < 0.01). Likewise, PSA secretion was increased by 56-fold in pTRE-5 alpha R1 transfectants treated with 10 pM testosterone, compared to 26-fold in pTRE transfectants (P < 0.01). At concentrations of testosterone above 10 pM, the stimulatory effect on growth and PSA secretion was not distinguishable between pTRE-5 alpha R1 and pTRE transfectants. CONCLUSIONS: These results demonstrate that upregulation of 5 alpha R1 enhances the cellular response to low, but not high, concentrations of testosterone. This explains one mechanism by which castration-recurrent PCa can proliferate in the presence of castrate levels of circulating testosterone. PMID- 19125421 TI - Adolescent female rats are more resistant than males to the effects of early stress on prefrontal cortex and impulsive behavior. AB - We tested the hypothesis that adolescent Sprague-Dawley females may be more resistant than males to display impulsive behavior and lower prefrontal cortex thickness after mother-infant separation (MS). Starting at postnatal day 2 (P2), the MS group was separated 6 hr/day and the early handled (EH) group 15 min/day for 10 days, and another group was standard facility reared (SFR). Subjects were examined for novel open-field activity (P28), light-dark apparatus (P29), familiar open-field (P30) and frontal cortical thickness. This protocol resulted in impulsive behavior in MS rats relative to EH and SFR, but this effect was less pronounced in females than males. MS affected the two sexes differently in terms of decreased prefrontal cortex dorsoventral thickness, with this effect being significant in males but not females. Neuroanatomical and behavioral documentation that adolescent females are more resistant than males to ADHD-like effects of maternal separation have not been previously reported. PMID- 19125424 TI - An analytical strategy for identification of a somatotropin-like bioactive peptide by ion trap liquid chromatography/electrospray ionization tandem mass spectrometry after immuno-affinity purification from buffalo serum. AB - The use of growth hormones, such as native and recombinant somatotropins, is forbidden in the European Union (EU), but is legal in the USA. The misuse of recombinant bovine somatotropin in Italy is suspected for enhancing milk production, thanks to its availability on the illegal market. A synthetic bioactive peptide of 27 amino acids derived from bovine somatotropin was successfully tested in France and in southern Italy for scientific purposes, to stimulate milk production, both in cows and buffaloes. This somatotropin-like peptide (PEP-ST), suspected for illegal use in southern Italy, was synthesized by linking the 104-113 sequence of bovine somatotropin to the 323-339 sequence of ovalbumin. Herein, a method for detection and identification of the PEP-ST in buffalo serum is described; our strategy was based on the production of IgG anti PEP-ST, used to synthesize an immuno-affinity column for peptide purification from buffalo serum, prior to analysis by ion trap liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). The immuno-affinity column was successfully used to purify in a single step the bioactive PEP-ST from buffalo serum samples spiked at 20, 50 and 200 microg/mL for confirmatory analysis. Ion trap LC/ESI-MS/MS identification was based on detection of a multi-charged molecular ion and its characteristic fragmentation pattern. No significant matrix interference was observed, accounting for method specificity. We consider this strategy to be a basic approach that could be improved in the perspective of the official control of illegal use of somatotropin and somatotropin-like compounds in buffalo breeding. PMID- 19125423 TI - Induction of apoptosis and nonsteroidal anti-inflammatory drug-activated gene 1 in pancreatic cancer cells by a glycyrrhetinic acid derivative. AB - Methyl 2-cyano-3,11-dioxo-18beta-olean-1,12-dien-30-oate (CDODA-Me) is a synthetic triterpenoid derived from glycyrrhetinic acid, a bioactive phytochemical in licorice, CDODA-Me inhibits growth of Panc1 and Panc28 pancreatic cancer cell lines and activates peroxisome proliferator-activated receptor gamma (PPARgamma)-dependent transactivation in these cells. CDODA-Me also induced p21 and p27 protein expression and downregulates cyclin D1; however, these responses were receptor-independent. CDODA-Me induced apoptosis in Panc1 and Panc28 cells, and this was accompanied by receptor-independent induction of the proapoptotic proteins early growth response-1 (Egr-1), nonsteroidal anti inflammatory drug-activated gene-1 (NAG-1), and activating transcription factor-3 (ATF3). Induction of NAG-1 and Egr-1 by CDODA-Me was dependent on activation of phosphatidylinositol-3-kinase (PI3-K) and/or p42 and p38 mitogen-activated protein kinase (MAPK) pathways but there were differences between Panc28 and Panc1 cells. Induction of NAG-1 in Panc28 cells was p38-MAPK- and PI3-K-dependent but Egr-1-independent, whereas induction in Panc1 cells was associated with activation of p38-MAPK, PI3-K, and p42-MAPK and was only partially Egr-1 dependent. This is the first report of the induction of the proapoptotic protein NAG-1 in pancreatic cancer cells. PMID- 19125425 TI - Interference of chlorofluorocarbon (CFC)-containing inhalers with measurements of volatile compounds using selected ion flow tube mass spectrometry. AB - Selected ion flow tube mass spectrometry (SIFT-MS) is a sensitive technique capable of measuring volatile compounds (VCs) in complex gas mixtures in real time; it is now being applied to breath analysis. We investigated the effect of inhalers containing chlorofluorocarbons (CFCs) on the detection and measurement of haloamines in human breath. SIFT-MS mass scans (MS) and selected ion monitoring (SIM) scans were performed on three healthy non-smoking volunteers before and after inhalation of the following medications: Combiventtrade mark metered-dose inhaler (MDI) (CFC-containing); Ventolintrade mark MDI (CFC-free); Atroventtrade mark MDI (CFC-free), Beclazonetrade mark MDI (CFC-containing); Duolintrade mark nebuliser. In addition, the duration of the persistence of the mass/charge ratios was measured for 20 h. Inhalers containing CFCs generated large peaks at m/z 85, 87, 101, 103 and 105 in vitro and in vivo, consistent with the predicted product ions of CFCs 12, 114 and 11. No such peaks were seen with Duolintrade mark via nebuliser, or CFC-free MDIs. We conclude that measurement of VCs, such as haloamines, with product ions of similar m/z values to the ions found for CFCs would be significantly affected by the presence of CFCs in inhalers. This issue needs to be accounted for prior to the measurement of VCs in breath in people using inhalers containing CFCs. PMID- 19125426 TI - Climatic factors influencing the isotope composition of Italian olive oils and geographic characterisation. AB - The purpose of this study was to investigate the possibility of identifying oil source areas by means of simple measurements on the natural samples avoiding time consuming sample treatments. The oxygen and carbon isotopic values of 150 samples of extra-virgin olive oil from eight different Italian regions and from three different years of production were measured according to well-established techniques. Statistical treatments of the results obtained show a very good correlation of the delta(18)O of oil with latitude, mean annual temperature, and mean relative humidity at the collection site. No correlation is found with elevation and mean annual precipitation. The shift of the oil delta(18)O per degree centigrade of the mean annual temperature is quantitatively close to that calculated for atmospheric precipitation in continental areas. Accordingly, in our measurements, the year of oil production can be identified on the basis of the delta(18)O value (mean 2004 temperatures were higher than 2005 temperatures). On the contrary, the oil delta(13)C values show no correlation with the above variables but only with latitude and, consequently, are less suitable for discriminating the geographic origin of oil. However, the delta(13)C values are suitable to indicate biological differentiation while the delta(18)O values are not. PMID- 19125427 TI - Elements of morphology: standard terminology for the periorbital region. AB - An international group of clinicians working in the field of dysmorphology has initiated the standardization of terms used to describe human morphology. The goals are to standardize these terms and reach consensus regarding their definitions. In this way, we will increase the utility of descriptions of the human phenotype and facilitate reliable comparisons of findings among patients. Discussions with other workers in dysmorphology and related fields, such as developmental biology and molecular genetics, will become more precise. Here we introduce the anatomy of the periorbital area and define and illustrate the terms that describe the major characteristics of the periorbital area. PMID- 19125428 TI - Elements of morphology: standard terminology for the lips, mouth, and oral region. AB - An international group of clinicians and scientists working in the field of dysmorphology has initiated the standardization of terms used to describe human morphology. The goals are to standardize these terms and reach consensus regarding their definitions. In this way, we will increase the utility of descriptions of the human phenotype and facilitate reliable comparisons of findings among patients. Discussions with other workers in dysmorphology and related fields, such as developmental biology and molecular genetics, will become more precise. Here we summarize the anatomy of the oral region and define and illustrate the terms that describe the major characteristics of the lips and mouth. PMID- 19125429 TI - Transmission mode ion/ion reactions in the radiofrequency-only ion guide of hybrid tandem mass spectrometers. AB - Transmission mode ion/ion reactions have been performed within the first quadrupole, the Q0 radiofrequency (RF)-only quadrupole, of two types of hybrid tandem mass spectrometers (viz., triple quadrupole/linear ion trap and QqTOF instruments). These transmission mode reactions involved the storage of either the reagent species and the transmission of the analyte species through the Q0 quadrupole for charge inversion reactions or the storage of the analyte ions and transmission of the reagent ions as in charge reduction experiments. A key advantage to the use of transmission mode ion/ion reactions is that they do not require any instrument hardware modifications to provide interactions of oppositely charged ions and can be implemented in any instrument that contains a quadrupole or linear ion trap. The focus of this work was to investigate the potential of using the RF-only quadrupole ion guide positioned prior to the first mass-resolving element in a tandem mass spectrometer for ion/ion reactions. Two types of exemplary experiments have been demonstrated. One involved a charge inversion reaction and the other involved a charge reduction reaction in conjunction with ion parking. Ion/ion reactions proved to be readily implemented in Q0 thereby adding significantly greater experimental flexibility in the use of ion/ion reaction experiments with hybrid tandem mass spectrometers. PMID- 19125430 TI - Stereochemical differentiation of C-7 hydroxyltaxane isomers by electrospray ionization mass spectrometry. AB - In this study, different electrospray ionization mass spectrometric (ESI-MS) methods were utilized to analyze several pairs of taxane stereoisomers including paclitaxel and 7-epi-paclitaxel. Both ESI-MS and tandem mass spectrometry (MS/MS) techniques provided stereochemically dependent mass spectra in negative-ion mode, and all studied stereoisomers could be easily distinguished based on their characteristic ions or distinct fragmentation patterns. MS/MS experiments for several taxane analogues at various collision energies were performed to elucidate potential dissociation pathways. The gas-phase deprotonation potentials were also calculated to estimate the most thermodynamically favorable deprotonation site using DFT B3LYP/6-31G(d). The results of the theoretical studies agreed well with the fragmentation patterns of paclitaxel and 7-epi paclitaxel observed from MS/MS experiments. In addition, it was found that liquid chromatography (LC)/ESI-MS was a useful and sensitive technique for assignment of C-7 taxane stereoisomers from realistic samples. PMID- 19125432 TI - Mass spectrometry in the characterization of ambers. II. Free succinic acid in fossil resins of different origin. AB - Baltic amber contains high levels of succinic acid, most of which is part of the polymer framework, and only recently has the presence of microcrystals of free succinic acid been documented by scanning electron microscopy and X-ray diffraction analysis. The determination of succinic acid can be used for an easy identification of Baltic amber among other fossil resins and to distinguish it from imitations, widely diffused on the market, produced with natural or modified recent or sub-fossil resins, such as copals. We report here a simple method, based on negative ion electrospray ionization mass spectrometry, for evaluating the free succinic acid content in water/methanol extracts of ambers from different origin, using a sample of about 20 mg. The limit of quantification is better than 1 ppm and, when applied to a set of amber samples of different origin, it was able to distinguish those of Baltic origin (showing free succinic acid levels in the range 50-400 ppm) from the others (for which succinic acid was undetectable, i.e. at levels lower than 1 ppm). PMID- 19125433 TI - Elements of morphology: standard terminology for the hands and feet. AB - An international group of clinicians working in the field of dysmorphology has initiated the standardization of terms used to describe human morphology. The goals are to standardize these terms and reach consensus regarding their definitions. In this way, we will increase the utility of descriptions of the human phenotype and facilitate reliable comparisons of findings among patients. Discussions with other workers in dysmorphology and related fields, such as developmental biology and molecular genetics, will become more precise. Here we introduce the anatomy of the hands and feet and define and illustrate the terms that describe the major characteristics of the hands and feet. PMID- 19125435 TI - Risk assessment in acute pancreatitis. AB - BACKGROUND: Acute pancreatitis has a variable natural history and in a proportion of patients is associated with severe complications and a significant risk of death. The various tools available for risk assessment in acute pancreatitis are reviewed. METHODS: Relevant medical literature from PubMed, Ovid, Embase, Web of Science and The Cochrane Library websites to May 2008 was reviewed. RESULTS AND CONCLUSION: Over the past 30 years several scoring systems have been developed to predict the severity of acute pancreatitis in the first 48-72 h. Biochemical and immunological markers, imaging modalities and novel predictive models may help identify patients at high risk of complications or death. Recently, there has been a recognition of the importance of the systemic inflammatory response syndrome and organ dysfunction. PMID- 19125434 TI - Timing and impact of infections in acute pancreatitis. AB - BACKGROUND: Although infected necrosis is an established cause of death in acute pancreatitis, the impact of bacteraemia and pneumonia is less certain. METHODS: This was a cohort study of 731 patients with a primary episode of acute pancreatitis in 2004-2007, including 296 patients involved in a randomized controlled trial to investigate the value of probiotic treatment in severe pancreatitis. Time of onset of bacteraemia, pneumonia, infected pancreatic necrosis, persistent organ failure and death were recorded. RESULTS: The initial infection in 173 patients was diagnosed a median of 8 (interquartile range 3-20) days after admission (infected necrosis, median day 26; bacteraemia/pneumonia, median day 7). Eighty per cent of 61 patients who died had an infection. In 154 patients with pancreatic parenchymal necrosis, bacteraemia was associated with increased risk of infected necrosis (65 versus 37.9 per cent; P = 0.002). In 98 patients with infected necrosis, bacteraemia was associated with higher mortality (40 versus 16 per cent; P = 0.014). In multivariable analysis, persistent organ failure (odds ratio (OR) 18.0), bacteraemia (OR 3.4) and age (OR 1.1) were associated with death. CONCLUSION: Infections occur early in acute pancreatitis, and have a significant impact on mortality, especially bacteraemia. Prophylactic strategies should focus on early intervention. PMID- 19125436 TI - Elements of morphology: standard terminology for the head and face. AB - An international group of clinicians working in the field of dysmorphology has initiated the standardization of terms used to describe human morphology. The goals are to standardize these terms and reach consensus regarding their definitions. In this way, we will increase the utility of descriptions of the human phenotype and facilitate reliable comparisons of findings among patients. Discussions with other workers in dysmorphology and related fields, such as developmental biology and molecular genetics, will become more precise. Here we introduce the anatomy of the craniofacies and define and illustrate the terms that describe the major characteristics of the cranium and face. PMID- 19125437 TI - The chemistry of ionized acetone/Ar mixtures under varying gas flow and mole ratio conditions: a matrix-isolation study. AB - The chemistry of ionized acetone:Ar mixtures under varying total gas flow rate and acetone:Ar mole ratio conditions has been studied using matrix-isolation techniques. Gaseous acetone diluted in excess argon gas was subjected to electron bombardment with 300-eV electrons. The products of subsequent reaction processes were matrix isolated and analyzed by Fourier transform infrared (FTIR) absorption spectroscopy. Products included 1-propen-2-ol (the enol isomer of acetone), methane, ketene, carbon monoxide, ethane, ethene, acetylene and tricarbon monoxide. Variations in the total flow rate of gas resulted in changes in the efficiency of product formation without significant changes in the relative amounts of the major species formed. Variations in the acetone:Ar mole ratio at fixed total gas flow resulted in striking variations in the products formed, demonstrating a shift from single acetone molecule-derived charge-transfer ionization chemistry at low acetone mole ratios, to processes consistent with the participation of two or more acetone molecules at intermediate mole ratios. These results are interpreted in the context of ion-molecule reaction processes, the onset of which occurs at intermediate acetone mole ratios. Ethane dehydrogenation products are proposed to result from product secondary ionization, a process that is prevalent at high ionizing electron fluxes. PMID- 19125438 TI - AIDS vaccine poised for human tests. PMID- 19125439 TI - Drug-resistant HIV strains turning up in China. PMID- 19125440 TI - US FDA expands approval for Johnson & Johnson HIV drug. PMID- 19125441 TI - Against disease: Impact of hygiene and cleanliness on health. PMID- 19125442 TI - Hydrophobic interaction capillary electrochromatography of protein mutants. Use of lipid-based liquid crystalline nanoparticles as pseudostationary phase. AB - Nanoparticle-based hydrophobic interaction-capillary electrochromatography was utilized for separation of proteins with similar mass-to-charge ratio at neutral pH without organic modifier. Lipid-based liquid crystalline nanoparticles were prepared and used as pseudostationary phase,benefiting from their high biocompatibility, ease of preparation,and suspension stability at high concentrations.Use of laser-induced fluorescence enabled detection at high nanoparticle concentrations. Green fluorescent protein(GFP) and mutants of GFP harboring single or double amino acid substitutions with the same charge were separated in the described system but not in conventional capillary electrophoresis. Separation was achieved by increasing the salt concentration to promote hydrophobic interactions by shielding of the repulsive electrostatic interactions. In addition, the method was adapted to a capillary with an effective length of 6.7 cm, enabling fast separations and future applications on chip. PMID- 19125443 TI - Text messages could hasten tuberculosis drug compliance. PMID- 19125444 TI - Alcohol use on the rise in India. PMID- 19125445 TI - Landscape and health. PMID- 19125446 TI - Atmospheric pressure free liquid infrared MALDI mass spectrometry: toward a combined ESI/ MALDI-liquid chromatography interface. AB - A new atmospheric pressure (AP)-MALDI-type interface has been developed based on a free liquid (FL) microbeam/microdroplets and a mid-infrared optical parametric oscillator (mid-IR OPO). The device is integrated into a standard on-line nanoESI interface. The generation of molecular ions in the gas phase is believed to be the result of a fast (explosive) laser-induced evaporative dispersion(not desorption) of the microbeam into statistically charged nanodroplets. Only the lowest charge states appear insignificant abundance in this type of experiment. Mass spectra of some common peptides have been acquired in positive ion mode, and the limit-of-detection of this first prototype (liquid microbeam setup) was evaluated to be 17 fmol per second. To improve the duty cycle and to reduce the sample consumption, a droplet-on-demand system was implemented (generating 100 pL droplets).With this setup, about 20 attomole of bradykinin were sufficient to achieve a signal-to-noise ratio better than five.This setup can be operated at flow rates down to 100 nL/min and represents a liquid MALDI alternative to the nanoESI. Our particular interest was the application of the developed ion source for on-line coupling of liquid chromatography with mass spectrometry. The flow rates(>100 microL/min), required for stable operation of the ion source in continuous liquid microbeam mode, matches perfectly the flow rate range of micro HPLC. Therefore, online LC/MS experiments have been realized, employing a microbore C18 reversed-phase column to separate an artificial peptide mixture and tryptic peptides of bovine serum albumin (performing a peptide mass fingerprint). In the latter case, sequence coverage of more than 90%has been achieved. PMID- 19125447 TI - Initial treatment of childhood chronic inflammatory demyelinating polyradiculoneuropathy. PMID- 19125448 TI - Electrokinetic detection and characterization of intact microorganisms. PMID- 19125449 TI - General approach for certain quantitative calculations for instance of the variance of reversible adsorption to the capillary wall in CE. AB - Miniaturization of analytical separation methods offers several advantages, including short run times, high resolution, and high recovery of the sample constituents. To optimize these parameters, the reversible adsorption (to minimize loss in resolution), as well as the irreversible adsorption (to minimize loss of analytes) must be quantified.However, no useful equation is available for the calculation of the variance of reversible adsorption. Therefore,we have taken another approach to quantify the reversible interaction. The method is unique and important since no equation for calculation of this variance is required. Instead, two experiments are required, which are run under such conditions that the variance of a certain parameter has the same numerical value in the two experiments (one with and without EOF), except for the variance of reversible adsorption. The approach is universal in the sense that it can be used for many different mathematical concepts and be modified to also cover certain functions other than a sum of parameters.We have also introduced a simple expression for their reversible adsorption, which shows that the hydrophobic interaction from only two methyl groups in the coating gives rise to as much as 40-50% loss of protein, and the width of the zones in the capillary with this coating was 8-15% larger compared to the zone width in the polyacrylamide-coated capillaries. The reproducibility in migration time, peak area, and peak width in two consecutive runs in capillaries with two methyl groups in the coating was very low, but in EOF-free polyacrylamide-coated capillaries extremely high, indicating that the reversible and irreversible adsorption of proteins to this coating is negligible. The scanning detector, frequently used in free zone electrophoresis in the 1960s 1970s, gives true separation parameters and is, therefore, much preferable to the stationary detector used in most CE experiments, because this detector gives apparent separation parameters. PMID- 19125451 TI - Potential and current density distributions at electrodes intended for bipolar patterning. AB - This paper deals with the use of reaction gradients on bipolar electrodes for the patterning of electrode surfaces.More specifically, the potential and current density distributions in two setups containing bipolar electrodes were investigated to optimize and design specific gradient geometries. Comparisons with simulations based on simple conductivity models showed a good qualitative agreement, demonstrating that these models could be used to predict bipolar behavior in more complex setups. In conjunction with imaging surface plasmon resonance(iSPR) experiments, the reaction gradients on bipolar electrodes could further be visualized. It was, for example,found that the gradient in potential difference was approximately linearly distributed in the center of the bipolar electrode and that these potential differences could be determined using an ordinary Ag/AgCl reference electrode.The present results thus provide a better understanding of the processes relevant for bipolar patterning.This approach was finally used to generate a circular gradient region in a self-assembled monolayer, thereby showing the possibilities to create interesting substrates for biosensors and microarray applications. PMID- 19125450 TI - Discovery of dopamine glucuronide in rat and mouse brain microdialysis samples using liquid chromatography tandem mass spectrometry. AB - A liquid chromatographic-electrospray/tandem mass spectrometric (LC-ESI-MS/MS) method was developed for the analysis of dopamine and its phase I and phase II metabolites from brain microdialysis samples. The method provides for the first time the analysis of intact dopamine glucuronide and sulfate without hydrolysis. The paper describes also an enzymatic synthesis method using rat liver microsomes as biocatalysts and characterization of dopamine glucuronide as a reference compound. The method was validated for quantitative analysis by determining limits of detection and quantitation, linearity,repeatability, and specificity. Dopamine glucuronide was found for the first time in rat and mouse brain microdialysis samples. The concentrations of dopamine and its glucuronide in the microdialysates collected from the striatum of rat brains were approximately equal (2 nM).Dopamine sulfate was not detected in the microdialysates(limit of detection 0.8 nM). The main metabolites of dopamine were dihydroxyphenylacetic acid (DOPAC,1200 nM) and homovanillic acid (HVA, 700 nM). PMID- 19125452 TI - What to do about social anxiety disorder. The holidays are coming, with lots of convivial occasions--fun for most, but distressing if you suffer from social phobia. PMID- 19125453 TI - Treating osteoporotic fractures of the spine. Several options, from pills to procedures, can help ease pain and shore up bone. PMID- 19125454 TI - Studies find a link between hot flashes and cardiovascular risk. PMID- 19125455 TI - Alexander technique helps relieve chronic back pain. PMID- 19125456 TI - By the way, doctor. Is there an age when a woman no longer needs a Pap smear? PMID- 19125457 TI - Urethral caruncle in a man. PMID- 19125458 TI - Delayed transvesical removal of a perforated tension-free vaginal tape, with maintenance of continence. PMID- 19125459 TI - Penile paraffinoma and a novel treatment. PMID- 19125460 TI - A lymph node metastasis from a 'burned-out' germ cell tumour presenting as an inguinal mass. PMID- 19125461 TI - De-inversion of an inverted appendectomy for the Malone procedure. PMID- 19125462 TI - Vesicolithiasis associated with posterior urethral valves. PMID- 19125463 TI - Chronic penoscrotal oedema: a presenting sign for Crohn's disease. PMID- 19125464 TI - High-flow priapism caused by injury of the bilateral cavernosal artery after needle trauma in a patient with low-flow priapism. PMID- 19125465 TI - Transient hyperparathyroidism after orthotopic bladder replacement. PMID- 19125466 TI - Squamous cell carcinoma of the renal pelvis: nephrostomy tract tumour seeding. PMID- 19125467 TI - Penile tuberculosis after intravesical bacillus Calmette-Guerin instillation. PMID- 19125468 TI - Pure testosterone-producing virilizing adrenocortical tumours in young women. PMID- 19125469 TI - Obturator nerve injury complicating a tension-free vaginal tape. PMID- 19125470 TI - Adult presentation of complete female urethral duplication. PMID- 19125471 TI - Radical prostatectomy for chronic prostatitis. PMID- 19125472 TI - An unusual cause of acute urinary retention in a child with pica disease. PMID- 19125473 TI - A novel mutation inducing partial androgen-insensitivity syndrome with phenotypic diversity. PMID- 19125474 TI - Laparoscopic port-site metastases from bladder transitional cell carcinoma. PMID- 19125475 TI - Primary urethral carcinoid tumour. PMID- 19125476 TI - [Problems in translation and terminology of disease names--special reference to the discussion at the Committee for the Psychiatric Terminology]. PMID- 19125477 TI - An unusual presentation of metastatic prostate cancer. PMID- 19125479 TI - Ureteric metastases from rectal carcinoma. PMID- 19125478 TI - Metastatic bacterial endophthalmitis from a prostatic abscess. PMID- 19125480 TI - [Estimation of degree of arousal with EEG]. PMID- 19125481 TI - Mullerianosis of the urinary bladder. PMID- 19125482 TI - Churg-Strauss syndrome and eosinophilic prostatitis. PMID- 19125483 TI - A novel way of controlling rectal bleeding after transrectal ultrasonography guided prostate biopsies. PMID- 19125484 TI - The use of aspirin with PC-SPES may not prevent pulmonary embolism. PMID- 19125485 TI - Squamous cell carcinoma in a male urethral diverticulum. PMID- 19125486 TI - Regulatory T cells in inflammatory bowel disease. AB - PURPOSE OF REVIEW: The intestinal immune system must orchestrate a complex balance between proinflammatory and anti-inflammatory responses to luminal antigens, and disruptions in this balance can result in inflammatory bowel disease (IBD). This review explores recent data that elucidate the role of regulatory T cells (Tregs) in the pathogenesis of IBD in mice and humans. RECENT FINDINGS: Data from murine models of colitis implicate several novel mechanisms critical to Treg function and generation including the inhibitory cytokine interleukin-35, pericellular adenosine generation and cytokine deprivation induced apoptosis. Although Tregs are essential in mice for the maintenance of intestinal homeostasis, their role in human IBD remains unclear. Patients with IBD appear to have relatively reduced numbers of Tregs in the blood and colon; however, Tregs from these patients are functional in vitro. SUMMARY: Tregs are important for the maintenance of intestinal self-tolerance and will likely prove to be an important avenue for therapeutic manipulation in IBD. PMID- 19125487 TI - Spontaneous regression of pulmonary metastases from renal cell carcinoma detected by computed tomography. PMID- 19125488 TI - A novel way to remove an usual foreign body in the urinary bladder. PMID- 19125489 TI - Necrotizing sarcoidosis of the kidney presenting as a renal mass. PMID- 19125490 TI - Spermatic cord lymphangioma in a 7-year-old child masquerading as a hydrocele. PMID- 19125491 TI - Life-threatening haemorrhage from an ileal conduit. PMID- 19125492 TI - Renal failure with inguinoscrotal herniation of the bladder. PMID- 19125493 TI - [Influence of the Japanese name change for schizophrenia]. PMID- 19125494 TI - [Patient's information. Upper digestive endoscopy]. PMID- 19125496 TI - Are you making your headaches worse? PMID- 19125495 TI - Don't let flying make you sick. PMID- 19125497 TI - [Epidemics and its responsive measures in Great Britain in 16th - 17th centuries]. AB - Historically, epidemics mean personal misery and social catastrophe in the history of humankind, and is another kind of living experience of the people. In 16th -17th century, confronting the epidemics, British society had various reactions and responsive measures, which were influenced by various factors, including medicine, religion, politics and economy. Though the religious beliefs had negative influence, to certain extent, on the reactions and medical development, the responsibility of the government was strengthened, replacing religious organization to undertake works in public health, demonstrating that this is an advancement. PMID- 19125498 TI - [The serious plague in Kaifeng in the third year of Chunhua of the Northern Song dynasty and the government's response]. AB - In the May of the third year of Chunhua (992) of the Northern Song dynasty, because of a long persistent drought and scorching hot day, the plague occurred and prevailed in Kaifeng, the capital of Northern Song, with a massive death toll, drawing much attention from the government. The cause of plague was related to high temperature, summer-heat warmth in TCM. In response to this, the government issued three imperial edicts: firstly, promulgating the formularies; secondly, sending some doctors to cure patient, as well as giving money and medicines. At the same time, the Imperial Medical Academy to preside over treatment and assigned an inner eunuch to be responsible for intendance; the third was sending the emissary to clear up the prison. This policies of the third year of Chunhua brought important effects to the system of prevention and rescue of epidemic disease in Song dynasty, exerting significant influences on the prevention and treatment of infectious diseases in the Song dynasty. PMID- 19125499 TI - [Changes of powder prescriptions in Qian jin yao fang (Essential Prescriptions Worth A Thousand Gold) made by Song people]. AB - As the contents in the two circulated versions of Qian jin yao fang (Essential Prescriptions Worth A Thousand Gold) ,i. e Bei ji qian jin yao fang (Essential Prescriptions Worth a Thousand Gold for Emergencies) and Xin diao sun zhen ren qian jin fang (Newly Printed Sun's Prescriptions Worth A Thousand Gold) are different greatly, especially on the literature of prescriptions. Song people made a lot of changes on the contents of powder prescriptions, including the related chapters and its contents, computation unit of materia medica, weights, and processing methods etc. without any remarks, nor notes in its preface. The contents in the Song version were more enriched than the newly carved version with the reasonable arrangement of its texts. As a clinical encyclopedia of traditional Chinese medicine, the Song version is more convenient for reading and using. However, it is a great taboo for the scholars to tamper an ancient work at will from bibliographic viewpoints. PMID- 19125500 TI - [Contributions of Shanghai physicians originated in Jiangsu province to traditional Chinese medicine in the Republic of China]. AB - The community of Shanghai physicians originated in Jiangsu province played an important role in the development of traditional Chinese medicine (TCM) during the period of the Republic of China. They acclimatized themselves to the social trend to establishing the medical schools, associations, journals and hospitals. For the survival rights of TCM, they led the TCM professionals to fight hard against those who, either in the society or in the government, wanted to abolish TCM and made outstanding contributions to the survival and further development of TCM. Their enterprising, innovative and indomitable spirit is our precious fortune needed to be inherited and developed. PMID- 19125501 TI - [Description of the relationship between body and mind in Kutadqu Bilik (wisdom for fortune and happy), a Uigur' s ancient book]. AB - The descriptions of the relationship between body and mind in Kutadqu Bilik ( Wisdom for Fortune and Happy) include the theory of five emotions (happiness, angriness, joy, annoyance and anxiety); the heart, liver and brain being the organs in charge of the function of the body and mind; and the viewpoint of spirit governing the body which, in turn, generates the spirit, and the overlapping of them. All of them have a scientific significance, forming the basis of the development of Uigur people and becoming a long-life ethnic group. PMID- 19125502 TI - [The research works on Shanghai Pasteur Institute]. AB - Shanghai Pasteur Institute, found in 1899, was the first institute of medical research on rabies with modern western medical science, and was also the first rabies vaccine manufacturer in China. By making use of relevant archives and documents, the authors attempt to elaborate its foundation and evolution, which focus on the fields of infectious disease prevention, popularization of vaccine inoculation, vaccine manufacturing and cultivation of Chinese medical talents. Before 1949, two branches of the Institute had been set up. It was incorporated into the newly established PLA Academy of Military Medicine Sciences in 1951. As an orient branch of the original Pasteur Institute, Shanghai Pasteur Institute inherit the sprit and methodology of the original Institute, and make great contribution to the foundation and development of Chinese modern microbiology, immunology and modern medical education. PMID- 19125503 TI - [Textual research on the modern circulation of Zhong jing quan shu (Zhongjing's Complete Works) block-printed edition in the Ming dynasty and collected in the library of China Medical University in modern times]. AB - The edition of Zhong jing quan shu (Zhongjing's Complete Works) recorded in the China Bibliography of Medical Books is different from the one in the Supplement to China Bibliography of Medical Books. The version of Zhong jing quan shu collected in the library of China Medical University is the Zhao Kai-mei's version of the Ming dynasty, printed after the Song version in Supplement to China Bibliography of Medical Books. This book was formally collected by the Manchuria Medical University on 8 November, 1936 (the 10th years of Japanese Showa). In 1945, Manchuria Medical University was taken over by the government of Kuomintang, and changed its name to National Shenyang Medical College. The Ministry of Public Health of Northeast People's government took over it in 1948, and merged into the China Medical University in June, 1949 together with its numerous rare ancient TCM books. All of them were borrowed by Liaoning College of Traditional Chinese Medicine in 1964. In March and April of 1970, these books were carried to Liaoyang, and then, to Shenyang in October. In 1978, the ancient books were retrieved by the China Medical University. Thus, the Zhao Kaimei's version of Ming dynasty of Zhongjing quan shu is preserved until today. PMID- 19125504 TI - [From medical intelligence journal to modern educational organ]. PMID- 19125505 TI - [25 years of progress in allergology: more potent weapons against aggressive allergens]. PMID- 19125506 TI - [25 years of progress in cardiology--cardiac arrhythmias: hot wires cure racing hearts]. PMID- 19125507 TI - [25 years of progress in cardiology--coronary heart disease: advances through self-experimentation and serendipity]. PMID- 19125508 TI - [25 years of progress in cardiology--heart failure: from haemodynamics to neuroendocrinology]. PMID- 19125509 TI - [25 years of progress in prevention: many more steps still must be taken]. PMID- 19125510 TI - [25 years of progress in hypertensiology: reducing blood pressure is not enough]. PMID- 19125511 TI - [25 years of progress in the treatment of metabolic disorders: statins revolutionise cardiovascular prevention]. PMID- 19125512 TI - [25 years of progress in diabetology: from strict regulations to flexible management]. PMID- 19125513 TI - [25 years of progress in oncology: we learned about chemotherapy at our patients' bedsides]. PMID- 19125514 TI - [25 years of progress in palliative care: creating space for a fulfilled life up to the very end]. PMID- 19125515 TI - [25 years of progress in pain therapy: the rocky road to recognition]. PMID- 19125516 TI - [High dosage stain protects heart and brain from recurrence]. PMID- 19125517 TI - ["No infarct without elevated LDL cholesterol"!]. PMID- 19125518 TI - [25 years of progress in HIV therapy: success story still lacks a happy end]. PMID- 19125519 TI - [25 years of progress in pharmacology: many innovations--are you up to date?]. PMID- 19125520 TI - [25 years of progress in paediatrics: a new type of morbidity is looming ahead]. PMID- 19125521 TI - [25 years of progress in gynaecology: significant advances in oncology and obstetrics]. PMID- 19125522 TI - [25 years of progress in urology: breakthrough for sexual dysfunction, testicular cancer and incontinence]. PMID- 19125523 TI - [The "stomach pathogen" can damage more than the stomach]. PMID- 19125524 TI - [Helicobacter pylori does not prevent asthma"]. PMID- 19125525 TI - [25 years of progress in dermatology: new therapies for skin diseases and alopecia]. PMID- 19125526 TI - [25 years of progress in ENT medicine: "deaf-mute" is no longer a definitive verdict]. PMID- 19125527 TI - [25 years of progress in neurology: an indispensable part of emergency medical care today]. PMID- 19125528 TI - [25 years of progress in psychiatry: we are heading in the right direction]. PMID- 19125529 TI - [25 years of progress in ophthalmology: preventing blindness is still a challenge]. PMID- 19125530 TI - [25 years of progress in orthopaedics: today's endoprostheses are safe and very durable]. PMID- 19125531 TI - [An alternative in microcirculation disorders. Homeopathic drug supports endothelial functions]. PMID- 19125532 TI - [25 years of progress in general and visceral surgery: from repair shop to fast track team]. PMID- 19125533 TI - [25 years of progress in forensic medicine: last witnesses: how maggots help solve cases of homicide]. PMID- 19125534 TI - Strengthening public health education in India: new initiatives. PMID- 19125535 TI - Efficient management of public health workforce. PMID- 19125536 TI - Continued use of asbestos: market's signature on science. PMID- 19125537 TI - An epidemiological study of low birth weight newborns in the district of Puruliya, West Bengal. AB - BACKGROUND: A community-based epidemiological study on birth weight of newborns was undertaken in the rural domain of a backward district of West Bengal. OBJECTIVES: To find out the magnitude of low birth weight (< 2.5 kg.) newborns and to highlight association, if any, of the socio-demographic, maternal and obstetric factors with birth weight of newborns. METHODS: The study was undertaken in 30 sub-centre areas (cluster), selected by '30 clusters sampling technique', in the rural area of Puruliya district. An adequate sample of 512 newborns (466 plus 10% extra allowance) delivered in those selected clusters (17 consecutive deliveries in each cluster) were to be studied. But, due to inability to record birth weight within 48 hours (in home deliveries) and other reasons, 487 newborns were included in assessing magnitude of the LBW problem. In-depth information on several variables was obtained from 439 mothers of the newborns. Health Workers (F), Anganwadi Workers, Trained dais and Health Supervisors -(F), specially trained for the survey, collected information by using pre-designed and pre-tested proforma. RESULTS & CONCLUSION: The incidence of LBW was 31.3%. Among different variables studied, statistically significant association was found in case of educational level of mothers and also place of delivery of newborns. More such community-based studies should be conducted PMID- 19125538 TI - Incidence of summer associated symptoms, host susceptibility and their effect on quality of life among women 18 to 40 years of age in an urban slum of Delhi. AB - OBJECTIVES: To study whether individual susceptibility plays a role in the occurrence of summer associated symptoms (dizziness, giddiness, fainting and weakness) among women 18-40 years of age and their effect on the quality of life. METHODS: It was a prospective community based study carried out in an urban slum of Delhi as a follow up of an earlier study. All women who had "given symptoms" during summer in the earlier study and a sample of those who were asymptomatic were included in the current study. Both the groups were studied for the occurrence of "given symptoms" and quality of life during next summer and winter using a pre structured questionnaire and WHOQOL-BREF. RESULTS: Incidence of "given symptoms" during summer was more than three times among women who were symptomatic earlier compared to women who were asymptomatic. The incidence of these symptoms was significantly higher among the former across different age groups and BMI categories. Their physical domain of quality of life was also adversely affected. CONCLUSIONS: Continued high incidence of summer associated symptoms with adverse effect on their physical quality of life, among women who were symptomatic earlier too, points to individual susceptibility rather than random occurrence. This aspect requires for further studies. PMID- 19125539 TI - Contemporary issues in public health governance--an Indian perspective. PMID- 19125540 TI - Climate change and health: methodological issues and introduction to climate epidemiology. AB - Climate change is a major threat to the health of current and future generations. This paper assesses the current estimations of adverse health outcomes of climate change and explores methodological issues. The study is essentially based on literature review. The estimation of disease burden does not portray an accurate picture of the problem due to methodological constraints on account of de-linking several non-climatic factors. In particular, the developing nations face greater challenges due to poor infrastructure, and lack of institutions and health services. The concept of climate epidemiology can minimize the existing research gap by adapting trans-disciplinary approach and it can establish as a new frontier for health advocacy in the larger context of global environmental change. PMID- 19125541 TI - Leptospirosis among patients with pyrexia of unknown origin in a hospital in Guwahati, Assam. AB - Leptospirosis is one of the important zoonoses of man and animals. In recent years, it has increasingly been reported from different parts of the world including India. Sera from 536 individuals with pyrexia of unknown origin (PUO) belonging to different age group, sex and occupation and having varying levels of exposure to animals, forest and flood were examined for the presence of antibodies to pathogenic Leptospira by commercially available kit. Overall 22.57% individuals with PUO were found to be positive for leptospirosis with 27.93% among male and 11.79% among female patients. This difference among..genders was statistically significant. The seropositivity was higher among patients in the age groups of 21-30 years followed by 31-40 and 41-50 years. Seropositivity was higher among individuals having contact with animals, working in forest and flood affected areas. PMID- 19125542 TI - Presence of food-borne microorganisms in milk in and around Guwahati city. PMID- 19125543 TI - Salmonella paratyphi A: an important cause of typhoid fever in Ludhiana. PMID- 19125544 TI - Direct toxicity assessment of volatile chlorinated hydrocarbon-contaminated groundwater and derivation of a site-specific guideline. AB - Groundwater contaminated with a mixture of 14 volatile chlorinated hydrocarbons (VCHs) discharges to an estuarine embayment in Sydney, Australia. A screening level hazard assessment identified a potential risk to aquatic organisms from surface water contaminated by the groundwater. Direct toxicity assessment of the groundwater was undertaken on 5 indigenous marine species to assess toxicity and derive a site-specific guideline. The testing included acute tests, subchronic tests on early life stages, and a chronic test. Test organisms included a microalga (Nitzschia closterium), an amphipod (Allorchestes compressa), a polychaete worm (Diopatra dentata), and sea urchin (Heliocidaris tuberculata) and oyster larvae (Saccostrea commercialis). Toxicity testing was undertaken in sealed containers to prevent loss of VCHs, and concentrations of VCHs were measured to accurately assess exposure concentrations. No observed effect concentration (NOEC) values varied from 1.56% dilution (1.11 mg total VCHs) to 50% dilution (45.5 mg total VCHs). EC50 values varied from 4.8% dilution (3.77 mg total VCHs) to more than 50% dilution (45.5 mg total VCHs). NOEC data were used to derive species sensitivity distributions (SSDs) and a site-specific guideline. SSDs were derived from Burr type III (including the Pareto) and log-normal distributions. The log-normal distribution represented the best fit, and because the Pareto distribution is a finite threshold model more suited to toxicants with a threshold mode of action, the log-normal SSD and the associated 95% trigger value (TV) of 830 microg/L of total VCHs, was adopted as the site-specific TV for the groundwater. PMID- 19125545 TI - Effects of atrazine on DNA damage and antioxidative enzymes in Vicia faba. AB - To evaluate atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) ecotoxicology in soil, the effect of atrazine on the activity of antioxidative enzymes (superoxide dismutase [SOD], catalase [CAT], and guaiacol peroxidase [POD]) was investigated in Vicia faba roots. Tissues from each treatment were collected on the days 7, 14, 21, and 28. Compared with the controls, SOD activity in V. faba roots was stimulated by the 2.5 mg/kg treatment and inhibited by the 5 and 10 mg/kg treatments, and CAT and POD activities in the 10 mg/kg treatment were inhibited on the whole. The Olive tail moments of single-cell gel electrophoresis of root cells were enhanced after treatment with different doses of atrazine on days 7, 14, 21, and 28, and significant differences were found compared to the controls. In conclusion, atrazine induces oxidative stress and DNA damage on V. faba. PMID- 19125546 TI - Phenanthrene metabolites bound to soil organic matter by birnessite following partial biodegradation. AB - The hypothesis that phenanthrene, an aromatic compound without a hydroxyl group, can form nonextractable residues in soil with the aid of phenanthrene biodegrading bacteria and birnessite was tested. The mutant strain Sphingobium yanoikuyae B8/36 successfully accumulated cis-phenanthrene dihydrodiol, and the intermediate was readily radicalized and coupled into soil organic matter in the presence of birnessite. Phenanthrene and the intermediate disappeared from the soil in 96 h in the presence of birnessite, but the intermediate accumulation occurred without birnessite. By determining the total organic carbon contents before and after birnessite treatment, it could be seen that birnessite did not mineralize cis-phenanthrene dihydrodiol. Fourier transform infrared and ultraviolet analyses suggest instead that the intermediate was incorporated into the soil organic matter, forming nonextractable, bound residues. Increases in the aromaticity and pH in birnessite-treated soil also present more evidence for bound residue formation. The soil in which bound residue formed did not exhibit an acute toxicity of phenanthrene, but evidence indicated that such toxicity existed in the freshly spiked soil. In addition, a long-term column test revealed that the bound residues could not be eluted by the combination of water, 80% methanol, and U.S. Environmental Protection Agency Toxicity Characteristic Leaching Procedure solution (pH 2.88) for four months, implying stability of the nonextractable residues in the soil. PMID- 19125547 TI - Analysis of pharmacist-provided medication therapy management (MTM) services in community pharmacies over 7 years. AB - BACKGROUND: Although community pharmacists have historically been paid primarily for drug distribution and dispensing services, medication therapy management (MTM) services evolved in the 1990s as a means for pharmacists and other providers to assist physicians and patients in managing clinical, service, and cost outcomes of drug therapy. The Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA 2003) and the subsequent implementation of Medicare Part D in January 2006 for the more than 20 million Medicare beneficiaries enrolled in the Part D benefit formalized MTM services for a subset of high-cost patients. Although Medicare Part D has provided a new opportunity for defining the value of pharmacist-provided MTM services in the health care system, few publications exist which quantify changes in the provision of pharmacist-provided MTM services over time. OBJECTIVES: To (a) describe the changes over a 7-year period in the primary types of MTM services provided by community pharmacies that have contracted with drug plan sponsors through an MTM administrative services company, and (b) quantify potential MTM-related cost savings based on pharmacists' self-assessments of the likely effects of their interventions on health care utilization. METHODS: Medication therapy management claims from a multistate MTM administrative services company were analyzed over the 7-year period from January 1, 2000, through December 31, 2006. Data extracted from each MTM claim included patient demographics (e.g., age and gender), the drug and type that triggered the intervention (e.g., drug therapeutic class and therapy type as either acute, intermittent, or chronic), and specific information about the service provided (e.g., Reason, Action, Result, and Estimated Cost Avoidance [ECA]). ECA values are derived from average national health care utilization costs, which are applied to pharmacist self-assessment of the "reasonable and foreseeable" outcome of the intervention. ECA values are updated annually for medical care inflation. RESULTS: From a database of nearly 100,000 MTM claims, a convenience sample of 50 plan sponsors was selected. After exclusion of claims with missing or potentially duplicate data, there were 76,148 claims for 23,798 patients from community pharmacy MTM providers in 47 states. Over the 7-year period from January 1, 2000, through December 31, 2006, the mean ([SD] median) pharmacy reimbursement was $8.44 ([$5.19] $7.00) per MTM service, and the mean ([SD] median) ECA was $93.78 ([$1,022.23] $5.00). During the 7-year period, pharmacist provided MTM interventions changed from primarily education and monitoring for new or changed prescription therapies to prescriber consultations regarding cost-efficacy management (Pearson chi-square P<0.001). Services also shifted from claims involving acute medications (e.g. penicillin antibiotics, macrolide antibiotics, and narcotic analgesics) to services involving chronic medications (e.g., lipid lowering agents, angiotensin converting enzyme [ACE] inhibitors, and beta-blockers; P<0.001), resulting in significant changes in the therapeutic classes associated with MTM claims and an increase in the proportion of older patients served (P<0.001). These trends resulted in higher pharmacy reimbursements and greater ECA per claim over time (P<0.001). CONCLUSION: MTM interventions over a 7-year period evolved from primarily the provision of patient education involving acute medications towards consultation-type services for chronic medications. These changes were associated with increases in reimbursement amounts and pharmacist-estimated cost savings. It is uncertain if this shift in service type is a result of clinical need, documentation requirements, or reimbursement opportunities. PMID- 19125548 TI - Evaluation of the first year of a pilot program in community pharmacy: HIV/AIDS medication therapy management for Medi-Cal beneficiaries. AB - BACKGROUND: The advent of combined antiretroviral therapy (ART) has increased treatment effectiveness but created new challenges for patients infected with human immunodeficiency virus (HIV) and for community pharmacists managing patients' drug therapy. The ability of pharmacist-provided medication therapy management (MTM) services to increase medication adherence, improve health outcomes, and reduce overall medical costs has been demonstrated in community pharmacies for chronic diseases such as diabetes and hypertension. However, the effectiveness of pharmacist-provided MTM services in HIV/acquired immune deficiency syndrome (AIDS) has not been well studied. In January 2005, a pilot program to evaluate MTM services for patients with HIV/AIDS began in California, allowing 10 HIV/AIDS specialty pharmacies to receive compensation for the MTM services that they provided to HIV/AIDS patients. OBJECTIVES: To examine the first year of the HIV/AIDS pharmacy MTM compensation pilot program, which described and compared pilot and nonpilot pharmacies with respect to (a) patient characteristics; (b) intermediate outcomes including type and number of ART medication regimens used, rates of adherence and excess medication fills for ART, use of contraindicated ART regimens, and occurrence of opportunistic infections; and (c) pharmacy and medical costs. METHODS: This was a cohort study examining 2005 Medi-Cal pharmacy and medical claims data for patients with HIV/AIDS who were served by pilot pharmacies versus other pharmacies. The HIV/AIDS patients were Medi-Cal beneficiaries aged 18 years or older as of January 1, 2005, who were continuously enrolled from January 1, 2004, through December 31, 2005, and diagnosed with HIV/AIDS, identified by receipt of at least 1 ART prescription and at least 1 medical claim with a diagnosis (primary or secondary) of HIV/AIDS (ICD 9-CM code 042.0) during both the index period (the year before pilot program implementation, 2004) and the intervention period (the study year, 2005). The only difference in the inclusion criteria for the 2 cohorts was that the pilot pharmacy patients were required to have filled 50% or more of their antiretroviral prescriptions in 2005 at 1 of the 10 pilot pharmacies. Adherence was defined as a medication possession ratio (MPR) of 80%-120% and excess medication fills as MPR greater than 120%. Comparisons were made between groups using bivariate statistics (Pearson chi-square for categorical variables and t tests for continuous variables). For comparisons of costs, generalized linear models assuming a gamma distribution and log link function were used; predictor variables for the models included age, gender, race/ethnicity, and dual coverage under Medicare. RESULTS: A total of 7,018 HIV/AIDS patients in the Medi-Cal population were identified as meeting the study criteria. Of these, 19.3% (n=1,353) were pilot pharmacy patients. The demographic profile of pilot pharmacy patients was similar, but not identical, to that of patients receiving medications at other pharmacies. A larger percentage of pilot pharmacy patients were on protease inhibitor-based ART medication regimens (63.8% vs. 54.8%, P<0.001), remained on a single type of ART therapy throughout the study year (56.8% vs. 34.2%, P<0.001), and were classified as adherent (56.3% vs. 38.1%, P<0.001), compared with other pharmacy patients. Fewer pilot pharmacy patients used contraindicated regimens (11.6% vs. 16.6%, P<0.001) or had excess medication fills (19.7% vs. 44.8%, P<0.001). The rate of opportunistic infections did not differ significantly between groups (28.2% vs. 26.1%, P=0.121). The total mean (standard error) annual health care cost per patient was 10% higher in pilot pharmacies than in other pharmacies ($40,596 [$889] vs. $36,937 [$479], P=0.001); driven by use of (a) medications (primarily non-ART medications) and (b) mental health services. Payment from the California Department of Health Care Services for MTM services averaged $1,014 per pilot pharmacy study patient. CONCLUSION: Study findings for the first year of the MTM program suggest that the pilot pharmacy patients received more appropriate HIV treatment. The degree to which these differences are affected by selfselection of patients into the pilot pharmacies is unknown. Longer-term outcomes and costs of the pilot program will be examined when data for subsequent years are available. PMID- 19125549 TI - Proton-pump inhibitor utilization associated with the change to nonpreferred formulary status for esomeprazole in the TRICARE formulary. AB - inhibitor (PPI) esomeprazole in the third copayment tier on the TRICARE formulary on July 17, 2005. The change to nonpreferred formulary status for esomeprazole included a $13 copayment increase (from $9.00 to $22.00) for either a 30-day supply purchased from a community pharmacy or a 90-day supply purchased from the mail-order pharmacy and a $0 copayment if obtained from a military pharmacy but with a prior authorization (PA) requirement. The change to nonpreferred formulary status was designed to encourage the use of PPIs other than esomeprazole and to increase the use of the mail-order pharmacy for esomeprazole purchases. OBJECTIVE: To quantify changes in (a) the TRICARE beneficiary utilization of esomeprazole relative to other PPIs and (b) the pharmacy settings used for filling esomeprazole prescriptions following implementation of a copayment increase and nonpreferred formulary status for esomeprazole. METHODS: A census of outpatient pharmacy fill records for prescription acid-reducing medications (PPIs, histamine-2 blockers, misoprostol, and sucralfate) obtained by beneficiaries aged 18 years or older from January 1, 2005, through December 31, 2006, was examined. Interrupted time series regression analyses without a control group were used to compare the utilization of esomeprazole relative to other PPIs, as well as the pharmacy setting used to obtain esomeprazole, in the months before and after the formulary change. The rates of continued esomeprazole use, switching to other prescription PPIs (lansoprazole, omeprazole, pantoprazole, and rabeprazole), switching to non-PPI prescription acid-reducing drugs, and discontinued prescription acid-reducing medication use among existing esomeprazole users (i.e., beneficiaries who obtained esomeprazole as the last PPI fill before the formulary change) were calculated overall and for each pharmacy setting used prior to the formulary change. RESULTS: Over the 24-month study period from January 1, 2005, through December 31, 2006, the total numbers of PPI fills and PPI users increased by 8.5% and 9.0%, respectively, and the number of esomeprazole users decreased by 4.6%. Of esomeprazole users, the percentages of individuals obtaining esomeprazole from military pharmacies and community pharmacies, respectively, decreased from 1.7% to 1.1% and from 89.7% to 81.7%, while the percentage of individuals obtaining esomeprazole from the mail-order pharmacy increased from 8.8% to 17.6%. Time series analysis yielded a positive, statistically significant growth in esomeprazole fills (beta1=0.114; P=0.012) during the 6-month pre-intervention period (January through June 2005) and a significant reduction in August 2005 (beta2=-5.0%; P<0.001), the month immediately following the formulary change. During the 17-month post-intervention period (August 2005 through December 2006), no statistically significant change in trend for esomeprazole fills (beta3=-0.0265; P=0.534) was observed, although a small increase in the raw number of esomeprazole fills was noted. Among the 117,801 existing esomeprazole users, 86,386 (73.3%) continued using esomeprazole, 17,676 (15.0%) switched to other prescription PPIs, 679 (0.6%) used only non- PPI prescription therapy, and 13,060 (11.1%) discontinued all prescription acid reducing pharmacotherapy after the formulary change. Significantly higher PPI switching and acid-reducing therapy discontinuation rates were observed among men aged 18-44 years and in TRICARE enrollees relative to women, those over 45 years of age, and those who used other health insurance (P<0.001). Individuals who used military pharmacies, where a PA requirement was implemented, were more likely to change pharmacy settings to obtain esomeprazole (43.8%) than were users of community pharmacies (11.9%) or the mail-order pharmacy (22.8%). Mail-order pharmacy users were less likely to discontinue acid-reducing pharmacotherapy (4.9%) than were community (11.9%) or military (12.9%) pharmacy users (P<0.05). CONCLUSIONS: After adjustment for serial correlation, the formulary change was associated with a migration of approximately 5% of all PPI fills and 25% of esomeprazole fills to the preferred PPIs in the first post-intervention month. Over the 17-month post-intervention period, the trend toward increased esomeprazole use was slowed and use of the mail-order pharmacy for esomeprazole fills nearly doubled. Our observed PPI switch rate of 15.0% resembled the rate observed for another insured population that experienced a similar formulary restructuring, but was substantially lower than the rates reported for more sizeable formulary changes. Thus, the present study's copayment differentials for third-tier medications ($19 compared with tier 1 and $13 compared with tier 2 copayments) may be less than the threshold amount required to optimize switching to preferred PPIs. PMID- 19125550 TI - Case study of the effects of office-based generic drug sampling on antibiotic drug costs and first-line antibiotic prescribing ratios. AB - BACKGROUND: Health plans and members benefit from the substitution of lower-cost drug therapies that achieve the same clinical outcomes as higher-cost drugs. Previous research suggests that generic sampling programs produce drug cost savings overall, but the effects attributable to acute therapies are unknown. Encouraging physicians to prescribe less expensive, first-line antibiotics may help reduce direct drug costs associated with prescribing potentially unnecessary, and more expensive, second-line agents. OBJECTIVES: To determine the effects of an automated, office-based generic drug sampling kiosk on (a) prescribing of first-line oral antibiotic agents as a ratio of total antibiotic prescribing and (b) average antibiotic drug cost per claim. METHODS: This managed care organization of 2.3 million members with pharmacy benefits collaborated with a vendor that developed an automated generic drug kiosk that allows for the dispensing of samples of generic medications within the prescriber's office. Among the samples contained in the kiosk were 6 generic, first-line oral antibiotics, representing 8 unique drug-strength options. Drug costs were defined as the ingredient cost of the drug claim, which includes plan cost, member cost share, and any dispensing fees or administrative program costs associated with the sampling program. In a difference-in-difference analysis, changes in outcome measures (antibiotic drug cost per claim and dispensing rates of first-line antibiotics) from 2003 (baseline year) to 2005 (post-implementation year) were compared among kiosk prescribers (n=179) and nonkiosk prescribers who were part of the same provider network (n=7,236). A cross-sectional analysis of the same outcome measures compared kiosk (n=396) and nonkiosk prescribers (n=10,267) in 2006. All statistical analyses were performed using t-tests of logtransformed data. RESULTS: The mean cost per claim dropped by $4.14 (12.3%) from $33.56 in 2003 to $29.42 in 2005 for the kiosk prescribers and by $3.35 (8.8%) from $38.26 in 2003 to $34.91 in 2005 for nonkiosk prescribers, but the mean change from 2003 to 2005 in the difference-in-difference analysis was not statistically significant (P=0.123). The first-line antibiotic prescribing ratio declined by 2.1 absolute points from 49.1% in 2003 to 47.0% in 2005 for the kiosk prescribers and by 3.4 points from 46.0% in 2003 to 42.6% in 2005 for the nonkiosk prescribers, but the difference- in-difference analysis showed that the change was not statistically significant (P=0.901). A cross-sectional analysis of 2006 data revealed significant differences between the kiosk prescribers versus their nonkiosk network counterparts for both first-line antibiotic prescribing rates (42.0% and 41.4%, respectively; P=0.028) and antibiotic cost per claim ($28.44 and $32.40, respectively; P<0.001). While the results of the cross-sectional analysis are statistically significant, the practical significance of the results is less evident. CONCLUSIONS: The benefits of including short-term medications such as first-line antibiotics in a generic drug sampling program are difficult to quantify, and the cost outcomes are unlikely on their own to justify generic drug sampling. However, acute (short-term) medications may be an effective part of an overall program to promote generic prescribing for all conditions. PMID- 19125551 TI - Resistance to in-office dispensing of generic antibiotic samples. PMID- 19125552 TI - Medicare part D: good for patients and an opportunity for pharmacists. PMID- 19125554 TI - There is value in anecdotal reports of relief from migraine with botulinum toxin. PMID- 19125553 TI - Prior authorization and clopidogrel use--the truth lies in the details. PMID- 19125555 TI - Medicare Part D-a roundtable discussion of current issues and trends. AB - BACKGROUND: Medicare Part D was introduced with a goal of providing access to prescription drug coverage for all Medicare beneficiaries. Regulatory mandates and the changing landscape of health care require continued evaluation of the state of the Part D benefit. OBJECTIVE: To review the current state of plan offerings and highlight key issues regarding the administration of the Part D benefit. SUMMARY: The Part D drug benefit continues to evolve. The benefit value appears to be diluted compared to the benefit value of large employer plans. Regulatory restrictions mandated by the Centers for Medicare and Medicaid Services (CMS) are reported to inhibit the ability of plans to create an effective, competitive drug benefit for Medicare beneficiaries. Management in this restrictive environment impedes competitive price negotiations and formulary coverage issues continue to create confusion especially for patients with chronic diseases. The doughnut hole coverage gap represents a significant cost-shifting issue for beneficiaries that may impact medication adherence and persistence. To address these and other challenges, CMS is working to improve the quality of care for Part D beneficiaries by designing and supporting demonstration projects. Although these projects are in different stages, all stakeholders are hopeful that they will lead to the development of best practices by plans to help manage their beneficiaries more efficiently. CONCLUSIONS: A significant number of Medicare beneficiaries are currently receiving prescription drug benefits through Part D. The true value of this benefit has been called into question as a result of plan design parameters that lead to cost-shifting, an increasing burden for enrollees. Concerns regarding the ability to provide a competitive plan given the stringent rules and regulations have been voiced by plan administrators. In an effort to drive toward evidence-based solutions, CMS is working to improve the overall quality of care through numerous demonstration projects. PMID- 19125556 TI - Health information technology--results from a roundtable discussion. AB - BACKGROUND: Health information technology (HIT), notably e-prescribing and electronic health records (EHR), have the potential to improve the quality of care, reduce medication errors and adverse events, and decrease overall health care utilization and costs. However, the United States continues to lag behind other countries in the adoption and use of HIT. OBJECTIVE: To review the various issues surrounding the implementation of HIT in the United States and potential drivers that will influence the use of e-prescribing and EHR. SUMMARY: The United States has been slow to embrace HIT. However, various factors, including increasing government involvement, are speeding the implementation and use of HIT. E-prescribing and EHR are both electronic means to provide better coordination of care by enabling various health care professionals to access patient medical records. Widespread adoption of HIT can be especially helpful for the elderly, since this population tends to have more chronic conditions requiring polypharmacy. Adoption of e-prescribing can reduce medication errors due to poor handwriting, while EHR can promote better clinical outcomes, improve medication adherence and refill rates, improve member satisfaction, and lower overall health care expenditures. Unfortunately, barriers to the adoption of e prescribing and EHR still exist, including resistance to learning new technology, initial start-up costs, delay in seeing a return on investment, lack of a standardized platform, increased administrative burden, and misaligned incentives. In an effort to promote greater adoption of e-prescribing and EHR, the Centers for Medicare & Medicaid has designed several initiatives, and other private organizations are now becoming more involved to close the HIT gap. CONCLUSION: Although the United States has been slow to implement HIT, there is reason to be hopeful. Increasing involvement by the government and other organizations will facilitate the greater adoption and use of e-prescribing and EHR in the near future. Ongoing data are needed, however, to demonstrate improvements in overall patient care and reductions in health care utilization and costs. These data are necessary to remove existing barriers that may prevent widespread implementation. PMID- 19125557 TI - The future of Medicare Part D drug plans--results from a roundtable discussion. AB - BACKGROUND: The Medicare Prescription Drug, Improvement, and Modernization Act, signed into law in 2003, provided access to prescription drugs for elderly Americans. The Part D benefit continues to evolve. Changes in plan designs, the impact of the doughnut hole on beneficiaries, and increased cost shifting have the potential to hamper the future of the Part D benefit. OBJECTIVE: To discuss factors that will likely have the most impact on the future of Medicare Part D from a patient and payer perspective. SUMMARY: The continued growth of the elderly population is expected to place an increasing burden on the services provided through Medicare. Given the current financial situation, it has been predicted that Medicare's Hospital Insurance Trust Fund will be depleted by 2019. To provide quality benefits and remain competitive, health plans are continually evaluating and redesigning their Part D benefits. However, the current regulatory environment is preventing plans from offering innovative products and designs that could lower costs to beneficiaries. The growing number of beneficiaries hitting the doughnut hole is also becoming a concern for both beneficiaries and health plans. More beneficiaries are reaching the doughnut hole, and this has resulted in changes in beneficiary behaviors, including stopping medications, switching to alternative drug classes, and reducing medication use. Because of the increasing concerns about Medicare's sustainability, it is anticipated that the government may become more involved. CONCLUSION: As the health care landscape continues to change, payers will be challenged to offer benefit designs that are affordable to elderly beneficiaries. For its part, the government must allow plans to design benefits that will improve the overall quality of care. Additionally, closer attention must be given to the growing number of beneficiaries hitting the doughnut hole and its potential adverse clinical and economic consequences. PMID- 19125558 TI - Rapid and reversible gel-sol transition of self-assembled gels induced by photoisomerization of dendritic azobenzenes. AB - An asymmetric bis-dendritic gelator (1) consisting of an azobenzene dendron and an aliphatic amide dendron was synthesized to achieve a photoresponsive self assembly. The compound gelled in a wide range of organic solvents, even at concentrations as low as 0.02% (w/v) in cyclohexane. The self-assembled fibrillar network structure was confirmed by field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM), and X-ray diffraction (XRD) analyses. The rapid and reversible gel-sol transition by irradiation with UV and visible light was investigated by UV-vis and Fourier transform infrared (FT-IR) spectroscopy, FE-SEM, and XRD analyses. Upon irradiation of the gel with UV, trans-to-cis isomerization of the azobenzene groups occurred, and the gel turned into a sol state. The gel was recovered immediately by the reverse cis-to-trans isomerization after the exposure to visible light. The trans-to-cis isomerization of the azobenzenes disrupted the hydrogen bonding of azobenzene amide groups, together with the hydrogen bonding in the aliphatic amide dendron. This facile communication between the two amide dendrons leads to the dissociation of the gel fibers and collapse of the gel. PMID- 19125559 TI - Preparation of monodisperse block copolymer vesicles via a thermotropic cylinder vesicle transition. AB - In aqueous solution, poly(2-vinylpyridine-b-ethylene oxide) spontaneously forms bilayer vesicles, the size of which can be tailored by extrusion through polycarbonate membranes. However, their size can be even more precisely influenced by subjecting them to a specific cooling/warming process proceeding through a cylinder-vesicle shape transition. The thermotropic alterations of the polymer aggregates and the topological pathways of the cylinder-vesicle transition were followed by dynamic light scattering (DLS) and cryo-electron microscopy (cryo-TEM). Upon cooling the vesicles to 4degreesC, there is a transition of the vesicles to basketlike aggregates and their further disintegration to wormlike micelles. Rewarming of the dispersion results in the reformation of vesicles via intermediate discoid and octopus-like structures. The variation of incubation times at 4 and 25degreesC, heating rate, polymer concentration, and ionic strength allows tailored preparation of unilamellar and almost monodisperse vesicles with diameters between 60 and 500 nm. Furthermore, fluorescently labeled dextrans, which were used as model drugs of differing molar mass, could be easily and stably encapsulated during the thermotropic formation of vesicles from wormlike micelles. PMID- 19125560 TI - Identification of adsorption sites in Cu-BTC by experimentation and molecular simulation. AB - The adsorption of several quadrupolar and nonpolar gases on the Metal Organic Framework Cu-BTC has been studied by combining experimental measurements and Monte Carlo simulations. Four main adsorption sites for this structure have been identified: site I close to the copper atoms, site I' in the bigger cavities, site II located in the small octahedral cages, and site III at the windows of the four open faces of the octahedral cage. Our simulations identify the octahedral cages (sites II and III) and the big cages (site I') as the preferred positions for adsorption, while site I, near the copper atoms, remains empty over the entire range of pressures analyzed due to its reduced accessibility. The occupation of the different sites for ethane and propane in Cu-BTC proceeds similarly as for methane, and shows small differences for O2 and N2 that can be attributed to the quadrupole moment of these molecules. Site II is filled predominantly for methane (the nonpolar molecule), whereas for N2, the occupation of II and I' can be considered almost equivalent. The molecular sitting for O2 shows an intermediate behavior between those observed for methane and for N2. The differences between simulated and experimental data at elevated temperatures for propane are tentatively attributed to a reversible change in the lattice parameters of Cu-BTC by dehydration and by temperature, blocking the accessibility to site III and reducing that to site I'. Adsorption parameters of the investigated molecules have been determined from the simulations. PMID- 19125561 TI - Self-assembled monolayers of alpha-cyclodextrin derivatives on gold and their host-guest behavior. AB - Various sulfur-modified alpha-cyclodextrin (alpha-CD) derivatives formed ordered monolayers on gold surfaces as confirmed by water contact angle goniometry, electrochemistry, X-ray photoelectron spectroscopy, and atomic force microscopy measurements. Self-assembled monolayers (SAMs) of the adsorbates showed high polarity, uniform monolayer arrangement, and low charge transfer resistance. Electrochemical capacitance measurements were used to determine the binding affinity of aliphatic carboxylic acid salts with four, six, and eight carbon atoms. The nonmethylated cyclodextrin host-guest pairs showed 1-2 orders of magnitude higher binding constants on surfaces than in solution. PMID- 19125562 TI - Surface segregation of fluorinated moieties on random copolymer films controlled by random-coil conformation of polymer chains in solution. AB - The relationship between solution properties, film-forming methods, and the solid surface structures of random copolymers composed of butyl methacrylate and dodecafluorheptyl methylacrylate (DFHMA) was investigated by contact angle measurements, X-ray photoelectron spectroscopy, sum frequency generation vibrational spectroscopy, and surface tension measurements. The results, based on thermodynamic considerations, demonstrated that the random copolymer chain conformation at the solution/air interface greatly affected the surface structure of the resulting film, thereby determining the surface segregation of fluorinated moieties on films obtained by various film-forming techniques. When the fluorinated monomer content of the copolymer solution was low, entropic forces dominated the interfacial structure, with the perfluoroalkyl groups unable to migrate to the solution/air interface and thus becoming buried in a random-coil chain conformation. When employing this copolymer solution for film preparation by spin-coating, the copolymer chains in solution were likely extended due to centrifugal forces, thereby weakening the entropy effect of the polymer chains. Consequently, this resulted in the segregation of the fluorinated moieties on the film surface. For the films prepared by casting, the perfluoroalkyl groups were, similar to those in solution, incapable of segregating at the film surface and were thus buried in the random-coil chains. When the copolymers contained a high content of DFHMA, the migration of perfluoroalkyl groups at the solution/air interface was controlled by enthalpic forces, and the perfluoroalkyl groups segregated at the surface of the film regardless of the film-forming technique. The aim of the present work was to obtain an enhanced understanding of the formation mechanism of the chemical structure on the surface of the polymer film, while demonstrating that film-forming methods may be used in practice to promote the segregation of fluorinated moieties on film surfaces. PMID- 19125563 TI - Quantitative profiling of polar cationic metabolites in human cerebrospinal fluid by reversed-phase nanoliquid chromatography/mass spectrometry. AB - Reversed-phase (RP) nanoliquid chromatography (LC)/mass spectrometry (MS) is widely used for proteome analysis, but hydrophilic metabolites are poorly retained on RP columns. We describe here the development and application of an efficient, robust, and quantitative nano-LC/MS method for cationic metabolome analysis in the positive ionization mode without any derivatization of analytes. Various stationary phases for nano-LC, coating of the internal wall of the capillary column, and various mobile phases were evaluated in terms of separation and peak shapes for 33 hydrophilic metabolites, including nonderivatized amino acids. Polar cationic compounds were strongly bound to mixed-functional RP with cation exchange mode resin, and the best separation was obtained with hydrophilic internal wall coating and a two-step trifluoroacetic acid (TFA) gradient in methanol as the mobile phase. Simple, but optimized, sample processing and the use of a high content of methanol allowed robust nano-LC/MS analysis. Our developed method was applied for biomarker discovery in Alzheimer's disease (AD). Several hundred peaks were detected from 10 microL of cerebrospinal fluid (CSF). In a principal component analysis (PCA) plot using peak intensities without normalization, peak separation depended on the experimental date, not disease state. Therefore, constant amounts of two stable isotope-labeled amino acids, Val and Lys, were added as internal standards (ISs) to each sample before processing. These ISs were eluted in different gradient slopes in the two-step gradient, and the normalized peak ratios using the corresponding ISs gave a unique group of PCA scores which could distinguish AD CSF samples from age-matched control CSF samples. PMID- 19125564 TI - Anion-controlled ion-pair recognition of paraquat by a bis(m-phenylene)-32-crown 10 derivative heteroditopic host. AB - It has been demonstrated that the complexation of the divalent salts of paraquat can be greatly improved (a K(a) increase up to 219 times was observed) by the introduction of ion-pair recognition through use of urea moieties on the crown ether. This improvement is controlled by not only the solvent polarity but also the nature of the anion. Furthermore, it was found that the binding motif for paraquat guest incorporation into the heteroditopic host is anion-controlled in the solid state. The host-guest complex is a pseudorotaxane in the solid state when the two counterions of paraquat are trifluoroacetate anions while it is a taco complex when the two counterions are hexafluorophosphate or chloride anions. PMID- 19125565 TI - Laser scanning up-conversion luminescence microscopy for imaging cells labeled with rare-earth nanophosphors. AB - Because of the ability to selectively reveal the objects of interest with subcellular resolution, fluorescence microscopy provides widespread applications from basic biological research to clinical diagnosis. However, challenges still remain in reducing the degree of photobleaching and increasing the contrast between signal and noise. Herein, we found that rare-earth nanophosphors exhibit a unique up-conversion luminescence mechanism and imaging modality and developed a new three-dimensional visualization method of laser scanning up-conversion luminescence microscopy (LSUCLM) with little photobleaching and no background fluorescence, by introducing a reverse excitation dichroic mirror and the confocal pinhole technique. Moreover, we demonstrated the up-conversion emission imaging of thin films containing embedded rare-earth nanophosphors and cells multilabeled with the nanophosphors and organic dyes. These data show that LSUCLM not only shares noninvasive benefits and deep penetration of two-photon microscopy but also offers some distinct advantages, such as little photobleaching of both organic dyes and rare-earth nanophosphors, no background fluorescence from either endogenous fluorophores or colabeled fluorescent probes, and excellent compatibility with conventional confocal microscopy. PMID- 19125566 TI - Salt effects on ion formation in desorption mass spectrometry: an investigation into the role of alkali chlorides on peak suppression in time-of-flight-secondary ion mass spectrometry. AB - In secondary ion mass spectrometry, the molecular environment from which a sample is analyzed can influence ion formation, affecting the resulting data. With the recent surge in studies involving examination of biological specimens, a better understanding of constituents commonly found in biological matrixes is necessary. In this article we discuss results from an investigation directed at understanding the role of salts doped as alkali chlorides in a model biological environment, arginine. The data show that addition of salt to the model system causes ion suppression of all the major mass spectral peaks attributed to arginine, with KCl having the largest suppression effect. Potential causes for the suppression effects are briefly discussed in relation to collected data. These theories include sample degradation, formation of salt adduct peaks, and anion neutralization. Investigation of the arginine salt data in comparison with data collected from pure salt systems indicates that suppression of the positive secondary ions is likely caused by a neutralization process involving the salt counteranion, chloride. To address the suppression issue, various procedures were performed on the arginine films such as sample washing with a cleaning solution (ammonium formate, ethanol, water) and analysis of films in a frozen-hydrated state. We present data from the analysis of the frozen-hydrated samples that shows both an ion yield enhancement and a significant amelioration of the salt suppression effects when compared to the samples run under standard conditions, demonstrating that it is a helpful approach to dealing with salt suppression. PMID- 19125567 TI - Differentiation of methyl-glucopyranoside anomers by infrared multiple photon dissociation with a tunable CO(2) laser. AB - Fragmentation of lithium cation-attached alpha- and beta-O-methyl-glucopyranoside precursor ions, formed by electrospray ionization (ESI) and trapped in a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer, was produced by infrared radiation from a tunable CO(2) laser. Irradiation over the wavelength range from 9.20 to 10.80 microm yielded unique fragmentation patterns that were a function of both product ion mass and laser wavelength. Comparison of the relative percent abundances of fragment ions for the two anomers revealed statistically significant differences for wavelengths between 9.20 and 9.70 microm at the 95% confidence level. On the basis of these results, differentiation of anomeric configurations of monosaccharides within oligosaccharides may be possible to address by wavelength-selective infrared multiple-photon dissociation (IRMPD). PMID- 19125568 TI - Enantioselective synthesis of benzimidazolyl quinoxalinones on soluble polymer support using focused microwave irradiation. AB - Focused microwave irradiation has been applied to a multistep synthetic sequence of reactions designed to generate benzimidazolyl quinoxalinones using a soluble polymer support. They were obtained by the ipso-fluoro (S(N)Ar) displacement of the immobilized ortho-nitro fluoro benzimidazoles with chiral alpha amino esters under microwave irradiation. Intermediate chiral organic-polymer conjugates when subjected to neutral reduction underwent a spontaneous intramolecular ring closure. Cleavage of the polymer support, at room temperature, did not cause any significant racemization resulting in the generation of a chiral molecular library with two points of structural diversity. PMID- 19125569 TI - Solid-phase synthesis and antitumor evaluation of 2,4-diamino-6-aryl-1,3,5 triazines. AB - 2,4-Diamino-6-aryl-1,3,5-triazines were synthesized by using a solid-supported approach in which monoarylsubstituted triazines were captured directly from the crude reaction mixture by resin-bound amines. The effects of the synthesized compounds on inhibition activities against tumor cell lines (PC-3, K562, A549, and HO8910) were examined. Most of the obtained compounds demonstrated remarkable antiproliferative activities against K562, PC-3, and HO8910 cell lines. Particularly, compounds 8c exhibited prominent inhibition activity with IC(50) values of 1.01, 2.23, and 1.06 microM, respectively. The structure-activity relationships of 2,4-diamino-6-aryl-1,3,5-triazines are also discussed. PMID- 19125570 TI - Dipole-induced, thermally stable lamellar structure by polar aromatic silane. AB - Controlled self-assembly of polar aromatic silane leads to the formation of well ordered lamellar structures. Graphite-like features are clearly visible with a scanning electron microscope (SEM). In addition, X-ray diffraction (XRD) patterns suggest a d spacing of 14.28 A along the z-axis and 4.42 A in the xy plane, which all agree with theoretical modeling. Constructing multistacks of silane molecules with a high degree of ordering is a daunting task. Amorphous monolayers are frequently reported. Aggravated van der Waals interaction, pi-pi electron overlapping, and solvophobic interactions can all lead to the formation of multistacks. The importance of a dipole to the ordered stacking is essentially unknown. This work suggests that a strong dipole-dipole interaction can be another important driving force in forming lamellar structures. The resulting large electrostatic interactions between the dipole and water provide an excellent thermal stability for these lamellas up to 350 degrees C. Organized, layered structures with a permanent dipole can be used in piezoelectric devices or as active surfaces to bind polar molecules, such as toxic gas, methanol, or DNA. PMID- 19125571 TI - Catalytic homologation of cycloalkanones with substituted diazomethanes. Mild and efficient single-step access to alpha-tertiary and alpha-quaternary carbonyl compounds. AB - Though volatile, toxic, and unstable, diazomethane is an indispensable one-carbon reagent with manifold uses in chemical synthesis. In this work, known protocols for hydrazone oxidation were adapted to permit facile access to a range of mono- and disubstituted aryl- or alkyldiazomethanes in pure form in solution; such procedures proceed in 30-60% overall yield starting from inexpensive carbonyl compounds. More important is the discovery that commercial Sc(III) salts are efficient catalysts for net insertion of the diazoalkyl carbon in these nucleophiles into the carbonyl C-C bond of simple cycloalkanones. In a single step, these reactions (1) forge two new C-C bonds under mild conditions, (2) produce molecular nitrogen as the sole stoichiometric byproduct, and (3) afford high yields of complex alpha-tertiary and -quaternary cyclic ketones that are typically accessible only through multistep procedures. PMID- 19125573 TI - Method to derive restoring forces of strained molecules from kinetic measurements. AB - We report a method to estimate the restoring force of a constrained small molecule and relate changes in its reactivity to this force. Restoring force is a size-invariant measure of molecular strain and offers a conceptual framework to bridge studies of macromolecular reactions by force spectroscopies with those of strained small molecules. We illustrate our method by showing that E stiff stilbene (1,1'-Delta-biindan) with up to 700 pN of force along its C6,C6' axis is accessible by photoisomerization. It reverts thermally to the Z isomer with tau(1/2) approximately 100 ms, probably long enough to be useful for microscale actuation. The quantum yield of photoisomerization and the activation barrier of thermal isomerization of constrained stiff stilbene increase sharply as the restoring force decreases. PMID- 19125575 TI - Label-free DNA biosensors based on functionalized carbon nanotube field effect transistors. AB - A carbon nanotube transistor array was used to detect DNA hybridization. A new approach to ensure specific adsorption of DNA to the nanotubes was developed. The polymer poly (methylmethacrylate(0.6)-co-poly(ethyleneglycol)methacrylate(0.15) co-N-succinimidyl methacrylate(0.25)) was synthesized and bonded noncovalently to the nanotube. Aminated single-strand DNA was then attached covalently to the polymer. After hybridization, statistically significant changes were observed in key transistor parameters. Hybridized DNA traps both electrons and holes, possibly caused by the charge-trapping nature of the base pairs. PMID- 19125574 TI - Influence of preformed Asp23-Lys28 salt bridge on the conformational fluctuations of monomers and dimers of Abeta peptides with implications for rates of fibril formation. AB - Recent experiments have shown that the congener Abeta(1-40)[D23-K28], in which the side chains of charged residues Asp23 and Lys28 are linked by a lactam bridge, forms amyloid fibrils that are structurally similar to the wild type (WT) Abeta peptide, but at a rate that is nearly 1000 times faster. We used all atom molecular dynamics simulations in explicit water, and two force fields, of the WT dimer, a monomer with the lactam bridge (Abeta(10-35)-lactam[D23-K28]), and the monomer and dimers with harmonically constrained D23-K28 salt bridge (Abeta(10 35)[D23-K28]) to understand the origin of the enhanced fibril rate formation. The simulations show that the assembly competent fibril-like monomer (N*) structure, which is present among the conformations sampled by the isolated monomer, with strand conformations in the residues spanning the N and C termini and a bend involving residues D(23) VGSNKG(29), are populated to a much greater extent in Abeta(10-35)[D23-K28] and Abeta(10-35)-lactam[D23-K28] than in the WT, which has negligible probability of forming N*. The salt bridge in N* of Abeta(10-35)[D23 K28], whose topology is similar to that found in the fibril, is hydrated. The reduction in the free energy barrier to fibril formation in Abeta(10-35)[D23-K28] and in Abeta(10-35)-lactam[D23-K28], compared to the WT, arises largely due to entropic restriction which enables the bend formation. A decrease in the entropy of the unfolded state and the lesser penalty for conformational rearrangement including the formation of the salt bridge in Abeta peptides with D23-K28 constraint results in a reduction in the kinetic barrier in the Abeta(1-40) lactam[D23-K28] congener compared to the WT. The decrease in the barrier, which is related to the free energy cost of forming a bend, is estimated to be in the range (4-7)k(B)T. Although a number of factors determine the growth of fibrils, the decrease in the free energy barrier, relative to the WT, to N* formation is a major factor in the rate enhancement in the fibril formation of Abeta(1-40)[D23 K28] congener. Qualitatively similar results were obtained using simulations of Abeta(9-40) peptides and various constructs related to the Abeta(10-35) systems that were probed using OPLS and CHARMM force fields. We hypothesize that mutations or other constraints that preferentially enhance the population of the N* species would speed up aggregation rates. Conversely, ligands that lock it in the fibril-like N* structure would prevent amyloid formation. PMID- 19125576 TI - Facile 1,3-diaza-Claisen rearrangements of tertiary allylic amines bearing an electron-deficient alkene. AB - Tertiary allylic amines with an electron-deficient alkene react with isocyanates and isothiocyanates to give highly substituted ureas and thioureas arising from formal 1,3-diaza-Claisen rearrangements. Isocyanates and isothiocyanates with strong electron-withdrawing groups are more reactive. Similarly, the data suggest that a stronger electron-withdrawing substituent on the alkene favors a faster reaction, but this may be offset by sterics in the cyclic transition state. PMID- 19125577 TI - Novel gel phase formed by mixing a cationic surfactive ionic liquid C(16)mimCl and an anionic surfactant SDS in aqueous solution. AB - The phase behavior of a catanionic system composed by a cationic surfactive ionic liquid (IL), 1-hexadecyl-3-methylimidazolium chloride ([C(16)mim]Cl), an anionic sodium dodecyl sulfate (SDS), and water has been investigated. A novel gel phase with quite high water content can be fabricated showing similar rheological properties to vesicles usually formed in traditional catanionic systems. The lamellar structure could also be constructed in SDS-rich region. Both the hydrophobic interaction of alkyl chains and interactions between oppositely charged head groups play important roles for the gel formation. Such a facile method to form gels directly from the catanionic system at relative low surfactant concentrations is novel, which should be related to the specific molecular structure of imidazolium ILs. The obtained results are expected to be helpful for better understanding of catanionic systems. PMID- 19125578 TI - Effect of organic manure on sorption and degradation of azoxystrobin in soil. AB - Information on pesticide degradation and factors influencing are important in predicting the levels of pesticide remaining in soils and allow assessment of potential risk associated with exposure. The present study reports the sorption and degradation of azoxystrobin [methyl (E)-2-{2-(6-(2-cyanophenoxy)pyrimidin-4 yloxy)phenyl}-3-methoxyacrylate] in a sandy loam soil. The fungicide was moderately sorbed, and the Freundlich adsorption parameter K(f) (1/n) values in natural and 5% compost-amended soils were 9.31 and 13.72, respectively. Sorption showed hysteresis with 32.5 and 14.7% of sorbed fungicide desorbed from the natural and 5% compost-amended soils, respectively. Azoxystrobin was more persistent in the aerobic soil than the anaerobic soil with half-life values of 107.47 and 62.69 days, respectively. Amendment of compost (5%) to the soil enhanced the degradation of fungicide, and the respective half-life values in aerobic and anaerobic soils were 73.39 and 38.58 days, respectively. Azoxystrobin acid was recovered as the only metabolite of azoxystrobin degradation in soils. Both sunlight and UV light affected the persistence of azoxystrobin with fungicide degraded at a faster rate in UV light than in sunlight. Soil acts as a screen and slows the fungicide degradation under sunlight and UV light. PMID- 19125579 TI - Design, synthesis, and cytoprotective effect of 2-aminothiazole analogues as potent poly(ADP-ribose) polymerase-1 inhibitors. AB - A series of novel poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors were designed within 2-aminothiazole analogues (4-10) based on a constructed three-dimensional pharmacophore model. After synthesis, the inhibitory effect on PARP-1 activity and the cytoprotective action of these compounds were tested and evaluated. Among them, compounds 4-6 and 10 appeared to be potent PARP-1 inhibitors with IC(50) values less than 1 microM, which had been perfectly predicted by pharmacophore model. These compounds proved to be highly potent against cell injury induced by H(2)O(2) and oxygen-glucose deprivation (OGD) in PC12 cells. These novel 2 aminothiazole analogues are potentially applicable as neuroprotective agents for the treatment of neurological diseases. PMID- 19125580 TI - A germanium(II) hydride as an effective reagent for hydrogermylation reactions. AB - Herein we report on the reactivity of the stable germanium(II) hydride LGeH (L = CH{(CMe)(2,6-iPr(2)C(6)H(3)N)}(2)) (2), which contains a low-valent germanium atom. 2 is prepared from the corresponding germanium(II) chloride LGeCl (1) using H(3)Al x NMe(3) or K[HB(iBu)(3)] in toluene. The reaction of 2 with carbon dioxide in toluene at room temperature affords a germanium(II) ester of formic acid, LGe-O-C(O)H (3), which is formed by insertion of the carbon dioxide into the germylene hydrogen bond. 2 also reacts with alkynes at room temperature to give the first germanium(II)-substituted alkenes (4, 5, and 6). These two reaction types have in common the fact that the hydrogen and germylene from LGeH are transferred to an unsaturated bond: the carbon-oxygen double bond (C=O) in the former case and the carbon-carbon triple bond (C[triple bond]C) in the latter. Moreover, the reaction of 2 with elemental sulfur in toluene at room temperature leads to the germanium dithiocarboxylic acid analogue LGe(S)SH (7). Compound 7 is formed by the unprecedented insertion of elemental sulfur into the germylene hydrogen bond and oxidative addition of elemental sulfur to the germanium(II) atom. This leads to the formal conversion of the GeH hydride to a SH proton. Compounds 3-7 were investigated by microanalysis, multinuclear NMR spectroscopy, and single-crystal X-ray structural analyses. PMID- 19125581 TI - Novel chiral molecular tweezer from (+)-usnic acid. AB - A new chiral molecular tweezer was synthesized with (1R,2R)-1,2 diaminocyclohexane as spacer and two molecules of (+)-usnic acid as pincers. The ability of this molecular tweezer to bind 2,4,7-trinitrofluorenone was studied. A charge-transfer complex was formed in which TNF was sandwiched between the two usnic acid units with pi-pi-stacked aromatic interactions. PMID- 19125582 TI - Reduction of a redox-active ligand drives switching in a Cu(I) pseudorotaxane by a bimolecular mechanism. AB - The reduction of a redox-active ligand is shown to drive reversible switching of a Cu(I) [2]pseudorotaxane ([2]PR(+)) into the reduced [3]pseudorotaxane ([3]PR(+)) by a bimolecular mechanism. The unreduced pseudorotaxanes [2]PR(+) and [3]PR(2+) are initially self-assembled from the binucleating ligand, 3,6-bis(5 methyl-2-pyridine)-1,2,4,5-tetrazine (Me(2)BPTZ), and a preformed copper macrocycle moiety (Cu-M(+)) based on 1,10-phenanthroline. X-ray crystallography revealed a syn geometry of the [3]PR(2+). The UV-vis-NIR spectra show low-energy metal-to-ligand charge-transfer transitions that red shift from 808 nm for [2]PR(+) to 1088 nm for [3]PR(2+). Quantitative analysis of the UV-vis-NIR titration shows the stepwise formation constants to be K(1) = 8.9 x 10(8) M(-1) and K(2) = 3.1 x 10(6) M(-1), indicative of negative cooperativity. The cyclic voltammetry (CV) and coulometry of Me(2)BPTZ, [2]PR(+), and [3]PR(2+) shows the one-electron reductions at E(1/2) = -0.96, -0.65, and -0.285 V, respectively, to be stabilized in a stepwise manner by each Cu(+) ion. CVs of [2]PR(+) show changes with scan rate consistent with an EC mechanism of supramolecular disproportionation after reduction: [2]PR(0) + [2]PR(+) = [3]PR(+) + Me(2)BPTZ(0) (K(D)*, k(d)). UV-vis-NIR spectroelectrochemistry was used to confirm the 1:1 product stoichiometry for [3]PR(+):Me(2)BPTZ. The driving force (DeltaG(D)* = 5.1 kcal mol(-1)) for the reaction is based on the enhanced stability of the reduced [3]PR(+) over reduced [2]PR(0) by 365 mV (8.4 kcal mol(-1)). Digital simulations of the CVs are consistent with a bimolecular pathway (k(d) = 12 000 s(-1) M(-1)). Confirmation of the mechanism provides a basis to extend this new switching modality to molecular machines. PMID- 19125583 TI - Phosphorylation site mapping of endogenous proteins: a combined MS and bioinformatics approach. AB - We present a novel approach that combines MALDI-TOF profile analysis and bioinformatics-based inclusion criteria to comprehensively predict phosphorylation sites on a single protein of interest from limiting sample. It is technologically difficult to unambiguously identify phosphorylated residues, as many physiologically important phosphorylation sites are of too low abundance in vivo to be unambiguously assigned by mass spectrometry. Conversely, phosphorylation site prediction algorithms, while increasingly accurate, nevertheless overestimate the number of phosphorylation sites. In this study, we show that MODICAS, an MS data management and analysis tool, can be effectively merged with the bioinformatics attributes of residue conservation and phosphosite prediction to generate a short list of putative phosphorylation sites that can be subsequently verified by additional methodologies such as phosphospecific antibodies or mutational analysis. Therefore, the combination of MODICAS driven MS data analysis with bioinformatics-based filtering represents a substantial increase in the ability to putatively identify physiologically relevant phosphosites from limited starting material. PMID- 19125584 TI - Identification and analysis of occludin phosphosites: a combined mass spectrometry and bioinformatics approach. AB - The molecular function of occludin, an integral membrane component of tight junctions, remains unclear. VEGF-induced phosphorylation sites were mapped on occludin by combining MS data analysis with bioinformatics. In vivo phosphorylation of Ser490 was validated and protein interaction studies combined with crystal structure analysis suggest that Ser490 phosphorylation attenuates the interaction between occludin and ZO-1. This study demonstrates that combining MS data and bioinformatics can successfully identify novel phosphorylation sites from limiting samples. PMID- 19125585 TI - Proteomics-based identification of differentially-expressed proteins including galectin-1 in the blood plasma of type 2 diabetic patients. AB - Type 2 diabetes (T2D) is a very heterogeneous and multifactorial disease. The pathophysiology of T2D is presumed to occur with an alteration in the levels of plasma proteins. To identify these differentially expressed proteins, plasma samples from normal and T2D humans were subjected to two-dimensional gel electrophoresis, quantitative densitometry, and mass spectrometry. Up to 200 protein spots were visible on each gel, of which 57 appeared modulated in diabetic individuals. Subsequently, 31 spots with > or =2-fold change in their expression were analyzed by MALDI-TOF mass spectrometry leading to the identification of 11 proteins with average sequence coverage of approximately 38%. The expression of apolipoprotein A-I was reduced by 4.2-fold, and galectin-1 was increased 4.8 times in diabetic samples. Induction of galectin-1 in T2D samples was confirmed by ELISA. In addition, the dose-dependent treatment of rat L6 skeletal muscle cells with glucose resulted in an upregulation of galectin-1. These data implicate the association of galectin-1 with the pathophysiology of diabetes and identify galectin-1 as a novel diagnostic marker protein in T2D patients. PMID- 19125587 TI - Efficient recovery of endothelial progenitors for clinical translation. AB - BACKGROUND: Circulating autologous CD133+ stem cells have been differentiated into a number of cell types that have the potential for clinical use, including endothelial cells. These cells are infrequently found in peripheral blood specimens, and this limits their use in clinical applications. To address this problem, we have developed an extracorporeal cellular affinity (ECA) column that can recover CD133 expressing progenitor cells with high efficiency. METHODS AND RESULTS: In sheep model, 1800 mL of blood was passed through a Sepharose-based column with affinity for CD133. Unbound cells and plasma were returned to the animal. Our results show that this process has a minimal effect on the hematologic and physiologic parameters of the animal. This recovery approach generated over 600-fold more endothelial colony forming units than conventional buffy leukocytes isolated from a peripheral blood specimen. Ultimately, the increased cell recovery of the ECA column enables the generation of a cell biomass for therapeutic purposes in nearly a third of the time. CONCLUSIONS: This technology may facilitate the generation of large numbers of progenitor-derived cells for clinical therapies and reduce the time required to attain clinically relevant cell numbers while minimizing loss of other important cell types to the donor. PMID- 19125593 TI - Fabrication, characterization, and optical properties of gold nanobowl submonolayer structures. AB - We report on a versatile method to fabricate hollow gold nanobowls and complex gold nanobowls (with a core) based on an ion milling and a vapor HF etching technique. Two different sized hollow gold nanobowls are fabricated by milling and etching submonolayers of gold nanoshells deposited on a substrate, and their sizes and morphologies are characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Optical properties of hollow gold nanobowls with different sizes are investigated experimentally and theoretically, showing highly tunable plasmon resonance ranging from the visible to the near-infrared region. Additionally, finite difference time domain (FDTD) calculations show an enhanced localized electromagnetic field around hollow gold nanobowl structures, which indicates a potential application in surface-enhanced Raman scattering (SERS) spectroscopy for biomolecular detection. Finally, we demonstrate the fabrication of complex gold nanobowls with a gold nanoparticle core which offers the capability to create plasmon hybridized nanostructures. PMID- 19125594 TI - Nanoparticle-containing membranes for the catalytic reduction of nitroaromatic compounds. AB - Layer-by-layer deposition of polyelectrolyte/metal nanoparticle films in porous alumina, track-etched polycarbonate, and nylon substrates yields catalytic membranes. With all three substrates, scanning electron microcopy images demonstrate a high density of well-separated nanoparticles in the membrane pores. These nanoparticles catalyze the reduction of nitroaromatic compounds by sodium borohydride with rate constants that are the same as those for nanoparticles immobilized on alumina powder. Moreover, the membranes selectively catalyze the reduction of nitro groups in compounds containing other reducible functionalities such as cyano, chloro, and styrenyl moieties. With nitrophenols and nitroanilines, the only reduction product is the corresponding amine. In contrast, nitrobenzene, nitrotoluenes, nitrobenzonitriles, chloronitrobenzenes, and m-nitrostyrene also form a nitroso product. Membrane catalysts are particularly attractive for controlling product distributions through variation of solution fluxes, as demonstrated by the formation of increased levels of nitroso compounds at high flux. PMID- 19125588 TI - Ibuprofen enhances recovery from spinal cord injury by limiting tissue loss and stimulating axonal growth. AB - The GTP-binding protein RhoA regulates microfilament dynamics in many cell types and mediates the inhibition of axonal regeneration by myelin and chondroitin sulfate proteoglycans. Unlike most other nonsteroidal anti-inflammatory drugs, ibuprofen suppresses basal RhoA activity (Zhou et al., 2003). A recent report suggested that ibuprofen promotes corticospinal axon regeneration after spinal cord injury (Fu et al., 2007). Here, we confirm that ibuprofen reduces ligand induced Rho signaling and myelin-induced inhibition of neurite outgrowth in vitro. Following 4 weeks of subcutaneous administration of ibuprofen, beginning 3 days after spinal cord contusion, animals recovered walking function to a greater degree, with twice as many rats achieving a hind limb weight-bearing status. We examined the relative role of tissue sparing, axonal sprouting, and axonal regeneration in the action of ibuprofen. Histologically, ibuprofen-treated animals display an increase in spared tissue without an alteration in astrocytic or microglial reaction. Ibuprofen increases axonal sprouting from serotonergic raphespinal axons, and from rostral corticospinal fibers in the injured spinal cord, but does not permit caudal corticospinal regeneration after spinal contusion. Treatment of mice with complete spinal cord transection demonstrates long-distance raphespinal axon regeneration in the presence of ibuprofen. Thus, administration of ibuprofen improves the recovery of rats from a clinically relevant spinal cord trauma by protecting tissue, stimulating axonal sprouting, and allowing a minor degree of raphespinal regeneration. PMID- 19125602 TI - Two-dimensional lamellar phase of poly(styrene sulfonate) adsorbed onto an oppositely charged lipid monolayer. AB - Polystyrene sulfonate (PSS 77 kDa) adsorbed onto oppositely charged dioctadecyldimethylammonium bromide (DODA) monolayers at the air/water interface is investigated with X-ray reflectivity and grazing incidence diffraction. The alkyl tails of DODA in the condensed phase form an oblique lattice with large tilts and intermediate azimuth angle. On PSS adsorption, the alkyl tail structure is maintained; only the tilt angle changes. Bragg peaks caused by flatly adsorbed, aligned PSS chains are observed, when DODA is in the fluid and also when it is in the condensed phase. The two-dimensional lamellar phase is only found at intermediate PSS bulk concentrations (0.001-1 mmol/L). In this phase, the PSS coverage can be varied by a factor of 3, depending on DODA molecular area and polymer bulk concentration. Charge compensation in the lamellar phase is almost achieved at 1 mmol/L. At larger bulk concentrations, PSS adsorbs flatly yet without chain alignment. Presumably, a necessary condition for a two dimensional lamellar phase is a pronounced electrostatic force which causes a large persistence length as well as repulsion between the aligned chains. PMID- 19125603 TI - Response of polyelectrolyte layers to the SiO2 substrate charging as probed by XPS. AB - A single layer of the cationic polyelectrolyte poly(allyamine) hydrochloride (PAH) deposited, using the layer-by-layer technique, on a silicon substrate containing 5 nm oxide layer is investigated by XPS while applying an external potential bias to the sample to control and manipulate the charge built-up on the oxide layer. Under application of a -10 V bias, the oxide layer is positively charged due to photoemission process, evidenced by the measured Si2p binding energy of 104.4 eV. Application of a +10 V bias attracts the low energy neutralizing electrons, stemming from a hot filament, and leads to a negatively charged oxide layer, also evidenced by the measured Si2p binding energy of 102.9 eV. The single polyelectrolyte overlayer also responds to this polarity change of the oxide layer underneath by displaying a somewhat larger shifts both in the C1s and N1s peaks. In addition to the shifts in the positions, the N1s peaks undergo a significant intensity depletion, mostly on the positively charged -N+ component. We interpret this intensity depletion to be the result of reorientation of some of the dangling positively charged groups by moving toward the negatively charged oxide underlayer. To our knowledge this is the first time that a chemically specific response to an electrical stimuli is reported using XPS. A bilayer LbL film consisting of PAH and PSS, exhibits even a larger charging shift, but this time no intensity alteration is observed, most probably due to locking of the -N+ groups by the -SO3+ counterions of the second layer. PMID- 19125608 TI - One-step formation of straight nanostripes from a mammal lipid-oleamide directly on highly oriented pyrolytic graphite. AB - Hierarchical nanostructures are obtained directly on highly oriented pyrolytic graphite (HOPG) by spin coating of dilute chloroform solution of 9-Z octadecenamide (oleamide), a natural lipid with cis-CdC- conformation, existing in the cerebrospinal fluid of mammal animals and being an additive for medical use and food packaging. Straight separated nanostripes with a length of 70-300 nm exist in the topmost layer and compact nanostripes in the bottom layer contacting HOPG. Compact nanostripes have a periodicity spacing of 3.8 nm, indicating H bonding between two rows of oleamide molecules. The orientation of the hierarchical nanostructures differs by n60 degrees+/-8 degrees (n=1 or 2), reflecting the epitaxial ordering along theHOPGsubstrate. The nanostripes are stable against annealing.Amolecular packing scheme for the nanostructures is proposed, where the -C=C bond angle in oleamide is 120 degrees and the plane of the carbon skeleton lies parallel to the HOPG substrate. Nanostripes in the topmost layer are formed from separated rows of oleamide molecules, due to the short-range surface potential of the substrate. The scheme involves direct influence ofHOPGon the orientation of oleamide molecules to form nanostripes without any purposely added saturated alkanes and H-bonds between amide groups in adjacent two rows of oleamide molecules. PMID- 19125609 TI - Meeting report: drug carriers in medicine and biology. PMID- 19125610 TI - Novel non-peptide nociceptin/orphanin FQ receptor agonist, 1-[1-(1 Methylcyclooctyl)-4-piperidinyl]-2-[(3R)-3-piperidinyl]-1H-benzimidazole: design, synthesis, and structure-activity relationship of oral receptor occupancy in the brain for orally potent antianxiety drug. AB - An endogenous heptadecapeptide, nociceptin/orphanin FQ (N/OFQ), and a G-protein coupled receptor, N/OFQ peptide (NOP) receptor [or opioid-receptor-like-1 (ORL1) receptor], have been described in terms of its structure, distribution, and pharmacology. Thus, the N/OFQ and NOP receptor are located in the central nervous systems in humans, primates, and rodents, and are involved in the integration of the emotional components in the brain; e.g., N/OFQ displays anxiolytic activity in the brain. For identifying orally potent anxiolytic, drug-design studies were performed with a series of 1,2-disubstituted benzimidazole derivatives, which resulted in the identification of various chemotypes of highly potent NOP selective full agonists in vitro with high or moderate NOP receptor occupancy in the mice brain per os such as 1-[1-(1-methylcyclooctyl)-4-piperidinyl]-2-[(3R)-3 piperidinyl]-1H-benzimidazole 1 (MCOPPB), the most potent novel non-peptide NOP full agonist in vitro and an orally potent anxiolytic in the mice. PMID- 19125611 TI - Redox modulation of Ca2+ signaling in human endothelial and smooth muscle cells in pre-eclampsia. AB - Pre-eclampsia (PE) is a leading cause of maternal hypertension in pregnancy and is associated with fetal growth restriction, premature birth, and fetal and maternal mortality. Activation and dysfunction of the maternal and fetal endothelium in PE appears to be a consequence of increased oxidative stress, resulting from elevated levels of circulating lipid peroxides. Accumulating evidence implicates reactive oxygen species (ROS) in the pathogenesis of vascular dysfunction in PE, perhaps involving a disturbance in intracellular Ca(2+) signaling. Several ion-transport pathways are highly sensitive to oxidative stress, and the resulting modulation of ion transport by ROS will affect intracellular Ca(2+) homeostasis. We review the evidence that changes in ion transport induced by ROS may be linked with abnormalities in Ca(2+)-mediated signal transduction, leading to endothelial and smooth muscle dysfunction in maternal and fetal circulations in PE. As dysregulation of Ca(2+) signaling in fetal umbilical endothelial cells is maintained in culture and embryonic, fetal, and postnatal development is affected by the cellular redox state, we hypothesize that impaired redox signaling in PE may influence "programming" of the fetal cardiovascular system and endothelial function in adulthood. PMID- 19125612 TI - Photodynamic therapy targets the mTOR signaling network in vitro and in vivo. AB - Mammalian target of rapamycin (mTOR) is a regulator of cell growth and proliferation and its activity is altered in many human cancers. The main objective of this study was to evaluate in vitro and in vivo targeting of mTOR by photodynamic therapy (PDT), a treatment modality for cancer. The amphiphilic endolysosomal localizing photosensitizer AlPcS(2a) and the p53 mutated rapamycin resistant colon adenocarcinoma cell line WiDr were used as models. AlPcS(2a)-PDT downregulated the levels of Ser(2448) phosphorylated mTOR (p-mTOR), total mTOR and phosphorylation of ribosomal S6 (p-S6) immediately after light exposure in a dose-dependent manner, indicating a direct targeting of the mTOR signaling network. Low-dose PDT attenuated the level of p-mTOR in a transient manner; approximately 35% reduction of p-mTOR was obtained 5 min after a LD(35) PDT dose, but returned to the basal level 24 h later. Treatment with the mTOR inhibitor rapamycin reduced the p-mTOR level by 25% after 4-24 h of incubation. Combination treatment of rapamycin and PDT in vitro resulted in synergistic cytotoxic effects when rapamycin was administered after PDT. However, antagonistic effects were obtained when rapamycin was incubated both before and after PDT. In vivo, activated mTOR in the WiDr-xenografts was downregulated by 35 and 75% 5 min and 24 h post PDT respectively as measured by immunoblotting. In contrast to untreated tumors where p-mTOR expression was found throughout the tumors, immunohistochemical staining revealed only expression of p-mTOR in the rim of the tumor at 24 and 48 h post PDT. In conclusion, AlPcS(2a)-PDT is a novel mTOR targeted cancer therapy. Rapamycin synergistically enhances the cytotoxicity of PDT only when administered post light exposure. PMID- 19125613 TI - Unusual structures and reactivity of mixed metal cluster complexes containing the palladium/platinum tri-t-butylphosphine grouping. AB - Polynuclear metal carbonyl complexes have a range of applications in chemical research: for example, they can serve as surface models to probe features of heterogeneous catalysis and can perform novel transformations of organic molecules in solutions. Mixed metal complexes can demonstrate bimetallic cooperativity and synergism and can also serve as precursors to multimetallic heterogeneous catalysts that have superior activities and selectivities. This Account describes the results of our recent comprehensive study of the chemistry of mixed metal cluster complexes containing the sterically encumbered M(PBu(t)(3)), M = Pd or Pt, group. This grouping readily adds to the metal-metal bonds of metal carbonyl cluster complexes and modifies their reactivity. We have prepared new, highly electronically unsaturated mixed metal complexes that exhibit unusually high reactivity toward hydrogen. The platinum atom of the Pt(PBu(t)(3)) grouping can bond to as many as five metal atoms, and it can interconvert, sometimes rapidly, between the different bonding modes. The large steric effects of the PBu(t)(3) ligand allowed us to prepare highly unsaturated, stable, mixed-metal complexes, and these complexes react with hydrogen, sometimes reversibly, under very mild conditions to yield polyhydride complexes. Strong evidence suggests that the Pt(PBu(t)(3)) group can also activate metal-hydrogen bonds in other complexes. In the future, we expect that researchers will prepare a greater variety of mixed metal complexes containing the Pd/Pt(PBu(t)(3)) group or other similar bulky groups, and that some of these complexes will exhibit even more unusual chemistry than what we have observed so far. PMID- 19125615 TI - Rat liver membrane glycoproteome: enrichment by phase partitioning and glycoprotein capture. AB - Past proteomic studies of membrane proteins have often been hampered by the low abundance and relatively high hydrophobicity of these proteins. Proteins are often glycosylated, particularly on the extracellular surface of the plasma membrane, and this characteristic was targeted as an enrichment strategy for identifying membrane proteins. Here, we report a strategy for identifying the tissue membrane glycoproteome, which involves (1) Triton X-114 phase partitioning, (2) isolation of glycosylphosphatidylinositol (GPI)-anchored proteins, and (3) glycoprotein capture using lectin affinity or hydrazine chemistry. Surprisingly, the capture of membrane proteins by lectin affinity and hydrazine chemistry resulted in mostly different populations of enriched glycoproteins. Lectins enriched high molecular weight functional membrane proteins with more potential glycosylation such as those involved in signal transduction and cell adhesion. Conversely, hydrazine chemistry isolated a higher proportion of smaller, enzymatic and peripheral membrane proteins such as solute carrier transporters and cytochrome p450s. We have applied our strategy to characterize the rat liver membrane glycoproteome and identified four new predicted GPI-anchored proteins and two that have not previously been seen in the liver. We also identified 424 nonredundant membrane proteins, of which 335 had potential N-linked glycosylation sites. PMID- 19125614 TI - Antigen delivery with poly(propylacrylic acid) conjugation enhances MHC-1 presentation and T-cell activation. AB - While many infectious diseases are controlled by vaccine strategies, important limitations continue to motivate the development of better antigen delivery systems. This study focuses on the use of a pH-sensitive polymeric carrier based on poly(propylacrylic acid) (PPAA) to address the need for more potent CD8 cytotoxic T-cell (CTL) responses. An MHC-1/CD8 CTL cell model system with ovalbumin as the protein antigen was used to test whether PPAA could enhance the delivery of ovalbumin into the MHC-1 display pathway. Ovalbumin was conjugated to poly(propylacrylic acid-co-pyridyldisulfide acrylate) (PPAA-PDSA) by disulfide exchange to make reversible conjugates that could be reduced by the glutathione redox system in the cytosol of antigen presenting cells. The PPAA-PDSA ovalbumin conjugates displayed the pH-sensitive membrane disruptive properties of the parent polymer as determined by their hemolysis activities (sharply active at the endosomal pH values of 6-6.5). The polymer-ovalbumin conjugates exhibited strong 22-fold increases in the MHC-1 presentation and ovalbumin-specific CTL activation compared to free ovalbumin. No CTL activation was observed with control conjugates of ovalbumin and poly(methylacrylic acid) (PMAA) that do not display membrane disruptive activies, suggesting that it is the membrane destabilizing properties of the polymer that result in increased MHC-1 display and CTL activation. Further mechanistic studies quantitated the time course of stable intracellular localization of radiolabeled conjugates. 52% of initially internalized PPAA-conjugated ovalbumin remained in the cells after 4 h, compared to less than 10% of ovalbumin or PMAA-ovalbumin. These results showing enhanced cytosolic delivery and MHC-1 presentation for the PPAA-antigen conjugates suggest that they warrant future characterization as a CD8-enhancing vaccine delivery system. PMID- 19125616 TI - Structure and activity of (2,8)-dicarba-(3,12)-cystino alpha-ImI, an alpha conotoxin containing a nonreducible cystine analogue. AB - The alpha-conotoxins are potent and selective antagonists of nicotinic acetylcholine receptors (nAChR). Exploitation of these and other peptides in research and clinical settings has been hampered by the lability of the disulfide bridges that are essential for toxin structure and activity. One solution to this problem is replacement of cystine bridges with nonreducible dicarba linkages. We explore this approach by determining the solution structure and functional characteristics of a dicarba analogue of the alpha-conotoxin alpha-ImI, (2,8) dicarba-(3,12)-cystino alpha-ImI. The structure of the dicarba analogue was similar to that of native alpha-ImI, with differences attributable to the different covalent geometry of the disulfide and dicarba bridges. Dicarba-alpha ImI maintained inhibitory activity of nAChR comparable to that of native alpha ImI in two in vitro assays. These findings confirm the potential of the dicarba linkage to improve stability while maintaining alpha-conotoxin function. PMID- 19125618 TI - First identification of 4-S-glutathionyl-4-methylpentan-2-one, a potential precursor of 4-mercapto-4-methylpentan-2-one, in Sauvignon Blanc juice. AB - The identification of 4-S-glutathionyl-4-methylpentan-2-one (glut-4-MMP) by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) experiments in a Sauvignon Blanc juice extract is described. Synthesis of an authentic reference compound enabled confirmation of the presence of glut-4-MMP in a Sauvignon Blanc juice for the first time. The juice extract was obtained by batch-wise percolation of 6 L of juice through a sintered glass funnel packed with C18 sorbent, followed by further purification using low-pressure chromatography on C18. Analysis of the juice extract revealed a chromatographic peak with the same retention time and mass spectrum as the synthetic reference compound, and spiking experiments verified the findings. The presence of glut-4 MMP in grape juice may be related to the biosynthesis of the relevant S-cysteinyl conjugate and, subsequently, to the formation of aroma-active 4-mercapto-4 methylpentan-2-one (4-MMP). This compound has a very low reported sensory threshold (3 ng/L) in wine and is partially responsible for the aromas that are important to the quality and style of some wine varieties. PMID- 19125619 TI - Pentaceno[2,3-b]thiophene, a hexacene analogue for organic thin film transistors. AB - Hexacene and larger fused rings remain elusive targets for chemists. Here, we report a hexacene-like molecule containing six linearly fused rings, specifically a pentacene molecule fused with a terminal thiophene ring, pentaceno[2,3 b]thiophene. It can be purified and isolated as a purple-black powder at ambient conditions. This molecule has a low HOMO-LUMO gap of 1.75 eV in o-DCB and an optical band gap of 1.58 eV in thin film. Top contact organic thin film transistors (OTFTs) were made, and atomic force microscopy (AFM) reveals a dendritic thin film growth characteristic of pentacene. An OTFT mobility of 0.574 cm(2)/V s was measured for pentaceno[2,3-b]thiophene under nitrogen. PMID- 19125621 TI - Sorption isotherms of ternary eluents in reversed-phase liquid chromatography. AB - The experimental technique of mass spectrometric tracer pulse chromatography was used to measure the excess volume of each eluent component for binary and ternary mixtures of water, acetonitrile, and methanol on a C(18)-bonded silica RPLC packing over the full composition range. The tracer pulse method allowed the direct measurement of excess volumes of each eluent component without numerical integration, assumed isotherm equation, detector calibration, or off-line analysis of the eluent composition. Absolute isotherms were estimated from the experimental data for excess volumes by use of various strategies for the estimation of the volumes of the stationary and mobile phases in dynamic equilibrium with eluents of varying composition. The results indicate that all three eluent components interacted with the alkane bonded phase. Some components were selectively taken up as part of the stationary phase while other components were selectively excluded so the composition of eluent in or on the stationary phase often significantly differed from the composition of the bulk eluent. The exact composition of the stationary phase (bonded phase plus immobilized eluent) was dependent upon the type and composition of the bulk eluent. PMID- 19125620 TI - Discovery of highly potent and selective inhibitors of neuronal nitric oxide synthase by fragment hopping. AB - Selective inhibition of neuronal nitric oxide synthase (nNOS) has been shown to prevent brain injury and is important for the treatment of various neurodegenerative disorders. This study shows that not only greater inhibitory potency and isozyme selectivity but more druglike properties can be achieved by fragment hopping. On the basis of the structure of lead molecule 6, fragment hopping effectively extracted the minimal pharmacophoric elements in the active site of nNOS for ligand hydrophobic and steric interactions and generated appropriate lipophilic fragments for lead optimization. More potent and selective inhibitors with better druglike properties were obtained within the design of 20 derivatives (compounds 7-26). Our structure-based inhibitor design for nNOS and SAR analysis reveal the robustness and efficiency of fragment hopping in lead discovery and structural optimization, which implicates a broad application of this approach to many other therapeutic targets for which known druglike small molecule modulators are still limited. PMID- 19125623 TI - Umbilical cord mesenchymal stem cells: role of regulatory genes in their differentiation to osteoblasts. AB - Umbilical cord (UC) mesenchymal stem cells (MSCs) are being currently investigated as an alternative to bone marrow (BM) MSCs for bone repair and regeneration. Here, we describe the gene regulation of their differentiation to osteogenic, adipogenic, and chondrogenic precursors and demonstrate their tendency to differentiate toward the osteoblast lineage. Fibroblast-like cells from the Warthon's Jelly were cultured with dedicated media to obtain osteogenic , adipogenic-, and chondrogenic-differentiated cells. After induction, a typical fibroblast-like shape with condensed fibers of F-actin was early noted in osteogenic-induced UC-MSCs, whereas those differentiating to adipocytes were flat with minor cytoskeleton relevance. Real-time PCR measured the transcription of master genes of the three lineages, thus revealing a remarkable up-regulation of Runx2 in osteogenic-induced cells with respect to both PPARg and SOX9 for adipogenic- and chondrogenic-differentiating UC-MSCs. However, TAZ, a coactivator of the nuclear transcription of Runx2 previously detected in BM-MSCs, was expressed in osteogenic- and, at lower magnitude, in adipogenic-induced cells, in keeping with its role in the reciprocal control of the differentiation between osteogenic- and adipogenic-induced cells. Its differential role in these cells was confirmed by its accumulation as protein product in the nuclei to activate Runx2 in osteogenic-differentiating UC-MSCs. These data emphasize the predominant expression by UC-MSCs of genes engaged in the osteogenic differentiation and their tendency to differentiate into osteoblasts, being similar in this respect to BM-MSCs. They may, thus, constitute a promising option for bone remodeling in regenerative medicine. PMID- 19125622 TI - Tubulin-interactive natural products as anticancer agents. AB - This review provides an overview of the discovery, structures, and biological activities of anticancer natural products that act by inhibiting or promoting the assembly of tubulin to microtubules. The emphasis is on providing recent information on those compounds in clinical use or in advanced clinical trials. The vinca alkaloids, the combretastatins, NPI-2358, the halichondrin B analogue eribulin, dolastatin 10, noscapine, hemiasterlin, and rhizoxin are discussed as tubulin polymerization inhibitors, while the taxanes and the epothilones are the major classes of tubulin polymerization promoters presented, with brief treatments of discodermolide, eleutherobin, and laulimalide. The challenges and future directions of tubulin-interactive natural products-based drug discovery programs are also discussed briefly. PMID- 19125624 TI - Mycangimycin, a polyene peroxide from a mutualist Streptomyces sp. AB - A mutualist actinomycete of the southern pine beetle, Dendroctonus frontalis, produces a polyene peroxide with pronounced antifungal activity. Its structure, absolute configuration, and biological activity were determined by spectral analysis, chemical modification followed by the modified Mosher method, and growth inhibitory assays, respectively. PMID- 19125626 TI - Regiocontrolled cobalt-catalyzed Diels-Alder reactions of silicon-functionalized, terminal, and internal alkynes. AB - The efficient control of the regiochemistry of the Diels-Alder adducts which are formed in excellent yields from 1,3-dienes and alkynylsilanes can be realized utilizing cobalt complexes with a pyridine-imine ligand or a dppe ligand, respectively. The application of 2-trimethylsilyloxy-1,3-butadiene leads to a very interesting cyclohexenone derivative suitable for further transformations. PMID- 19125625 TI - Cascade palladium-catalyzed alkenyl aminocarbonylation/ intramolecular aryl amidation: an annulative synthesis of 2-quinolones. AB - Palladium-catalyzed intermolecular aminocarbonylation/intramolecular amidation cascade sequences can be used to convert a range of 2-(2-haloalkenyl)aryl halide substrates efficiently and selectively to the corresponding 2-quinolones. Delaying the introduction of the CO atmosphere allows an amination/carbonylation sequence and the preparation of an isoquinolone. PMID- 19125628 TI - Inductive transfer of knowledge: application of multi-task learning and feature net approaches to model tissue-air partition coefficients. AB - Two inductive knowledge transfer approaches - multitask learning (MTL) and Feature Net (FN) - have been used to build predictive neural networks (ASNN) and PLS models for 11 types of tissue-air partition coefficients (TAPC). Unlike conventional single-task learning (STL) modeling focused only on a single target property without any relations to other properties, in the framework of inductive transfer approach, the individual models are viewed as nodes in the network of interrelated models built in parallel (MTL) or sequentially (FN). It has been demonstrated that MTL and FN techniques are extremely useful in structure property modeling on small and structurally diverse data sets, when conventional STL modeling is unable to produce any predictive model. The predictive STL individual models were obtained for 4 out of 11 TAPC, whereas application of inductive knowledge transfer techniques resulted in models for 9 TAPC. Differences in prediction performances of the models as a function of the machine learning method, and of the number of properties simultaneously involved in the learning, has been discussed. PMID- 19125629 TI - Reversal of diabetes in rats using GLP-1-expressing adult pancreatic duct-like precursor cells transformed from acinar to ductal cells. AB - Pancreatic injury induces replacement of exocrine acinar cells with ductal cells. These ductal cells have the potential to regenerate the pancreas, but their origin still remains unknown. It has been reported that adult pancreatic acinar cells have the potential to transdifferentiate to ductal progenitor cells. In this regards, we established novel adult pancreatic duct-like progenitor cell lines YGIC4 and YGIC5 and assessed the usefulness of these ductal progenitors in the cell therapy of diabetic rats. Acinar cells were cultured from pancreata of male Sprague Dawley rats and gradually attained ductal cell characteristics, such as expression of CK19 and CFTR with a concomitant down-regulation of amylase expression over time, suggesting transdifferentiation from acinar to ductal cells. During cell culture, the expression of Pdx-1, c-Kit, and vimentin peaked and then decreased, suggesting that transdifferentiation recapitulated embryogenesis. Overexpression of pancreas development regulatory genes and CK19, as well as the ability to differentiate into insulin-producing cells, suggests that the YGIC5 cells had characteristics of pancreatic progenitor cells. Finally, YGIC5 cells coexpressing Green fluorescent protein (GFP) and glucagon-like peptide (GLP)-1 under the activation of a zinc-inducible metallothionein promoter were intravenously infused to STZ-induced diabetic rats. Hyperglycemia was ameliorated with elevation of plasma insulin, and GFP-positive donor cells were colocalized in the acinar and islet areas of recipient pancreata following zinc treatment. In conclusion, after establishing pancreatic progenitor cell lines YGIC4 and YGIC5 under the concept of acinar to ductal transdifferentiation in vitro, we demonstrate how these adult pancreatic stem/progenitor cells can be used to regulate adult pancreatic differentiation toward developing therapy for pancreatic disease such as diabetes mellitus. PMID- 19125630 TI - A new isotherm for multisite occupancy adsorption of binary gaseous mixtures. AB - Adsorption is one of the most popular methods for reducing pollutants or separation of gases. Therefore it is important to introduce new and extended isotherm equations for binary systems. In the previous adsorption isotherms for binary systems, it is assumed that an adsorbate occupies one site in lattice sites. However, the adspecies may occupy more than one site in the lattice. Here we propose an adsorption isotherm for binary systems where the adsorbates occupy more than one site. The new isotherm equation for multisite occupancy of binary mixtures was derived based on statistical thermodynamics. The present new adsorption isotherm provides information about the structure of molecules in adsorbed form. Finally, the results of the present theoretical study were confirmed by analysis of two experimental systems. PMID- 19125631 TI - Neutron-scattering probe of complexes of sodium dodecyl sulfate and serum albumin during polyacrylamide gel electrophoresis. AB - Small-angle neutron scattering (SANS) is used to probe the conformation of SDS BSA protein surfactant complexes during electrophoresis in cross-linked polyacrylamide gels. Contrast variation permits independent probing of the structure of protein-surfactant complexes with negligible scattering contributions from the polyacrylamide matrix. The conformation of the protein complexes in the gel is found to be independent of the electric fields that are applied in this work (10 V/cm). Furthermore, there are no signs of large-scale macromolecular orientation (anisotropy) in the scattering patterns. However, the scattering shows that there are significant interparticle correlations between the protein-surfactant complexes that are electrophoretically inserted into the gel. These interactions develop when the total concentration of protein in the gels reaches values that are larger than approximately 1 mg/mL. The correlations are due to molecular crowding in the small fraction of pores that are available for protein migration. PMID- 19125632 TI - Water mediates the metastable crystal-to-stable crystal phase transition process in phospholipid aqueous dispersion. AB - The metastable crystal usually appears first during crystallization from solution, and it will then transform into the stable form. This metastable crystal-to-stable crystal phase transition process is important in the field of crystal growth and engineering. Crystallization behaviors of amphiphiles such as phospholipids are special as they have preassembled structures prior to crystallization, and the fundamental mechanisms regarding the transformation details of their polymorphic forms at the molecular level are not well understood. In this work, by using Fourier transform infrared spectroscopy combined with the hydrogen/deuterium (H/D) exchange method, we have demonstrated that water plays a critical role in mediating the metastable crystal-to-stable crystal phase transition process of a representative phospholipid, dilauroylphosphoethanolamine (DLPE). The H/D exchange method applied here is a powerful and simple approach to probe if water is involved in the crystal-to crystal transformation process. The results indicate that water works like a catalyst for the physical process. Furthermore, the conformational changes of the lipid molecules during the crystal-to-crystal transition process have been examined. Understanding the details of the transition from a metastable crystal to a stable crystal will shed light on the crystal growth theory and engineering of the self-assembled amphiphiles in aqueous solutions. PMID- 19125633 TI - Computer simulations of vesicle fission induced by external amphipathic inclusions. AB - Fission is an important biological phenomenon to maintain normal functions of cells, but its mechanism is not quite clear. In this paper, the fission behaviors of a two-component lipid vesicle induced by external amphipathic inclusions are studied by the dissipative particle dynamics method. We find that the fission of the lipid domain the in a spherical vesicle will occur only when the concentration of amphipathic inclusions is beyond a threshold concentration. It reveals that this kind of fission is induced by the combination of the domain's line tension and the asymmetric distribution of the amphipathic inclusions in the membrane's inner and outer leaflets. Furthermore, we also find that the fission behaviors are influenced by the area-to-volume ratio of vesicles and the structure characters of amphipathic inclusions (e.g., chain length and rigidity). Especially, the chain length and rigidity of amphipathic inclusions have complicated effects on the vesicle fission. With the variation of these factors, the fission process will be accelerated or delayed. The present study is helpful to understand the possible fission mechanism of lipid bilayers with embedded proteins. PMID- 19125634 TI - Detection of quadruplex DNA by luminescence enhancement of lanthanide ions and energy transfer from lanthanide chelates. AB - Small amounts of quadruplex DNA have been detected using luminescence enhancement of aqueous lanthanide ions and energy transfer from lanthanide chelates. The 22mer human telomeric DNA, AGGG(TTAGGG)(3), was detected using europium ions at concentrations as low as 20 ppb DNA. Detection with terbium ions was not possible due to the inherent weak luminescence intensities of lanthanides. Two different terbium chelates were used to overcome this challenge. When quadruplex DNA was added to the chelates there was a change in the excited-state lifetime of the chelate with subsequent energy transfer to the DNA. Experiments showed an increase in the amount of energy transferred from the chelate to the human telomeric DNA and other quadruplex sequences increased as a function of DNA concentration. PMID- 19125635 TI - H-bond-assisted regioselective (cis-1) intramolecular nucleophilic addition of the hydroxyl group to [60]fullerene. AB - The one-step reaction of 2,6-dihydroxyphenylmethyl ketone and sarcosine with [60]fullerene in refluxing chlorobenzene affords, in a totally regioselective process, the cis-1 bicyclic-fused organofullerene through a new intramolecular nucleophilic addition of one hydroxy group to the fullerene double bond. Experimental findings reveal the presence of a methyl group on C-2 of the pyrrolidine ring as an essential requirement for the cyclization process, whereas the existence of a H-bond between a second hydroxylic group and the nitrogen atom of the pyrrolidine ring seems to favor the approaching geometry without determining the reaction outcome. Theoretical calculations using the two-layered ONIOM approach and density functional theory support the experimental findings, predicting the strong impact that the presence of the methyl substituent on the C 2 of the pyrrolidine ring has on the molecular geometry and, hence, on the intramolecular cyclization process. PMID- 19125636 TI - Adaptation of a membrane-active peptide to heterogeneous environment. II. The role of mosaic nature of the membrane surface. AB - In the first article of this series we demonstrated the importance of specific intrapeptide interactions and peptide-lipid contacts for the membrane binding of penetratin (pAntp). Here in focus was detailed characterization of spatial hydrophobic/hydrophilic properties of the bilayer surface and their influence on the binding mode of pAntp. From the hydrophobicity point of view, the solvent accessible surfaces of lipid bilayers possess a distinctly "mosaic" character. This correlates well with the occurrence of dynamic clusters of hydrophobic surface area formed by hydrocarbon tails of phospholipids exposed on the interface. Such mosaic patterns are specific for lipid bilayers of particular composition. In an anionic membrane, they determine initial stages of pAntp adsorption, which strongly depends on the "complementarity" between polarity properties of the peptide and its local interfacial environment. If high complementarity is established, then pAntp penetrates deeply into the membrane without significant destabilization of its initial secondary structure. Alternatively, partial unfolding of pAntp takes place in order to compensate unfavorable peptide-membrane interactions upon embedding. Such effects explain complicated behavior of membrane-active peptides, especially if the target membrane surface is of distinctly mosaic nature, depending on the microscopic properties of the water-lipid interface, pAntp is capable of adopting different pathways to exercise its biological activity. PMID- 19125637 TI - Graft protection in bypass surgery: siRNA-mediated silencing of adhesion molecules. AB - The outcome of patients after coronary bypass grafting is greatly influenced by the type of graft material employed, especially regarding the rate of graft restenosis. Besides direct thrombotic events, the leukocyte-endothelial interaction modulated by adhesion molecules is identified to be the central cause leading to graft alterations. This study deals with a new therapeutic concept in order to achieve superior protection of a new bypass graft by blocking the adhesion molecule expression pathway with RNA interference to inhibit the initial leukocyte adhesion and transmigration. Leukocyte binding to adhesion molecules on activated human venous endothelial cells (HVECs) was determined by video-assisted microscopy in a flow chamber mimicking physiological conditions. The cells under study were sequentially transfected in a nonviral manner with specific short interfering RNA-sequences (siRNA) targeting E-selectin, intercellular adhesion molecule, and vascular adhesion molecule. After stimulation of adhesion molecule expression by tumor necrosis factor, a leukocyte-rich suspension was run through the chamber and the attaching leukocytes were counted. Transfection with specific siRNA targeting three different adhesion molecules resulted in a highly significant reduction of leukocyte attachment to activated HVECs in each case compared to the controls (p < 0.05). Transfection with a mixture out of all three siRNA-sequences showed the lowest leukocyte adhesion (p < 0.05) compared to the controls. siRNA-sequences inhibit the adhesion molecule expression on HVECs in an extremely effective way; not only in a single transfection of specific molecules but also in a parallel transfection of multiple sequences in one transfection. Accordingly, siRNA treatment significantly reduced adhesion of leukocyte cells to HVECs compared to controls. This study showed for the first time an effective knockdown of the leukocyte-endothelium interactions by transfection of HVECs with a cocktail consisting of three highly specific siRNAs against three different endothelial adhesion molecules. PMID- 19125638 TI - Inhibition of human cancer-cell proliferation by long double-stranded RNAs. AB - Three different enzymatically synthesized long double-stranded RNAs (dsRNAs) [448 bp homologous to the third exon of c-myc messenger RNA (mRNA) (dsMyc); 473 bp homologous to enhanced green fluorescent protein (EGFP) mRNA (dsEGFP) and control interferon inducer poly(I:C)] were studied for antiproliferative and gene silencing activities in KB-3-1, SK-N-MC, and IMR-32 human cancer cell lines. Simple incubation with these dsRNAs did not affect the expression of c-myc gene and the proliferation of KB-3-1 and IMR-32 cells, but inhibited the proliferation of SK-N-MC cells. Transfection of KB-3-1 and SK-N-MC cells using Oligofectamine dsRNAs complexes resulted in dose-dependent inhibition of c-myc and beta-actin genes expression and proliferation. The data show that dsMyc, acting both as interferon inducer and as gene-specific interfering RNA, is more effective as c myc inhibitor than other tested dsRNAs. The most efficient inhibition of proliferation was displayed by dsEGFP RNA, dsMyc and poly(I:C) were effective only when used in higher concentrations. Our data indicate that transfection of studied dsRNAs causes an increase in apoptotic and dead cells number in the cell population. This proapoptotic activity correlates with dsRNAs-induced antiproliferative activity. However the difference in cell growth between dsRNA treated and Oligofectamine-only treated cells can not be attributed only to the loss of cells due to the apoptosis; it also indicates some retardation of cell cycle progression caused by dsRNA. PMID- 19125639 TI - Therapeutic potential of splice-switching oligonucleotides. AB - Alternative splicing enables a single pre-messenger RNA transcript to yield multiple protein isoforms, making it a major contributor to the diversity of the proteome. While this process is essential for normal development, aberrations in alternative splicing are the cause of a multitude of human diseases. Methods for manipulating alternative splicing would thus be of therapeutic value. Chemically modified antisense oligonucleotides that alter alternative splicing by directing splice site selection have been developed to achieve this end. These splice switching oligonucleotides (SSOs) have been applied to correct aberrant splicing, induce expression of a therapeutic splice variant, or induce expression of a novel therapeutic splice variant in a number of disease-relevant genes. Recently, in vivo efficacy of SSOs has been reported using animal disease models, as well as in results from the first clinical trial. PMID- 19125640 TI - Adaptation of a membrane-active peptide to heterogeneous environment. I. Structural plasticity of the peptide. AB - Some membrane-active peptides undergo drastic changes of conformation and/or orientation on water-lipid interfaces. Among the most notable examples is penetratin (pAntp), a short cell-penetrating peptide. To delineate the driving forces behind pAntp-membrane interactions, we used, in this series of two papers, a combined modeling approach that includes: (1) molecular dynamics simulations of pAntp in zwitterionic and anionic lipid bilayers, (2) free energy perturbation calculations of model residue-residue contacts, and (3) detailed analysis of spatial hydrophobic/hydrophilic properties of the peptide/membrane systems. In this first article, we consider the role of conformational plasticity of the peptide in different membrane surroundings, as well as the ability of pAntp to form stable specific residue-residue interactions and make contacts with particular lipids. It was shown that pAntp displays a complicated conformational behavior. Basic and aromatic residues of the peptide form energetically favorable pairs in water and apolar environments, which facilitate membrane insertion of the peptide and stabilization of the membrane-bound state. These residues are also capable of "trapping" lipid heads, thereby affecting their dynamics and microscopic organization of the water-lipid interface. The latter effect is much more pronounced in anionic bilayers and might be related to the initial stage of peptide-induced destabilization of lipid bilayers. PMID- 19125641 TI - Encapsulated pheochromocytoma cells secrete potent noncatecholamine factors. AB - Pheochromocytomas are widely believed to induce cardiomyopathy via hypersecretion of catecholamines, including norepinephrine (NE). NE can have direct cardiomyocyte toxicity and/or can stimulate myocardial remodeling secondary to the induction of hypertension. Yet, the development of cardiomyopathy is not entirely related to catecholamine dose or the extent of hypertension. To explore these effects, we engineered a polymeric encapsulation system to control PC12 cell kinetics and NE release in vitro and in vivo. Primary neonatal rat cardiomyocytes incubated with pheochromocytoma-conditioned media exhibited greater cytoskeletal changes than myocytes cultured with identical doses of NE alone, including more profound dose-dependent decreases in desmin, beta-tubulin, and vinculin and upregulation of dystrophin. Cardiomyocyte contractility was 29 +/- 6% greater at given levels of NE release. Agarose-encapsulated PC12 cells retain cell viability and structural integrity in vivo. These implants induce a 30% greater degree of cardiac enlargement as compared to pumps releasing equivalent doses of NE. Protein level alterations observed in vitro were mirrored in vivo after implantation of encapsulated cells or NE pumps for 28 days. Together, these data suggest that pheochromocytoma-induced cardiomyopathy is not solely a catecholamine-mediated event; rather, the pathogenesis of this dilated cardiomyopathy appears to be dependent upon secondary factors unexamined to date. PMID- 19125642 TI - Telomerase immortalized human amnion- and adipose-derived mesenchymal stem cells: maintenance of differentiation and immunomodulatory characteristics. AB - Cell banking of mesenchymal stem cells (SCs) from various human tissues has significantly increased the feasibility of SC-based therapies. Sources such as adipose tissue and amnion offer outstanding possibilities for allogeneic transplantation due to their high differentiation potential and their ability to modulate immune reaction. Limitations, however, concern the reduced replicative potential as a result of progressive telomere erosion, which hampers scaleable production and long-term analysis of these cells. Here we report the establishment and characterization of two human amnion-derived and two human adipose-derived SC lines immortalized by ectopic expression of the catalytic subunit of human telomerase (hTERT). hTERT overexpression resulted in continuously growing SC lines that were largely unaltered concerning surface marker profile, morphology, karyotype, and immunosuppressive capacity with similar or enhanced differentiation potential for up to 87 population doublings. While all generated lines showed equal immunomodulation compared to the parental cells, one of the amnion-derived immortalized lines resulted in significantly increased immunogenicity. Although telomerase proves as important tool for immortalizing cells, our data emphasize the need for careful and standardized characterization of each individual cell population for cell banks. PMID- 19125643 TI - Tissue engineering of feline corneal endothelium using a devitalized human cornea as carrier. AB - The difficulties in obtaining good quality tissue for the replacement of corneas of patients suffering from endothelial dysfunctions have prompted us to evaluate the feasibility of producing a tissue-engineered (TE) corneal endothelium using devitalized human stromal carriers. Thus, corneal substitutes were produced by seeding cultured feline corneal endothelial cells on top of previously frozen human corneal stromas. After two weeks of culture to allow attachment and spreading of the seeded cells, the TE corneal endothelium was stained with alizarin red for endothelial cell count and fixed for histology, immunofluorescence labeling, scanning and transmission electron microscopy. Histology and Hoechst staining showed that there were no remaining cells in the devitalized stroma. After seeding, histology and transmission electron microscopy showed that the TE corneal endothelium formed a monolayer of tightly packed cells that were well adhered to Descemet's membrane. Scanning electron microscopy corroborated that the cells covered the entire posterior corneal surface and had an endothelial morphology. Alizarin staining showed that mean cell counts were 2272 +/- 344 cells/mm(2), indicating that the cell density was appropriate for grafting. The TE feline corneal endothelium also expressed the function-related proteins Na(+)/HCO(3)(-), ZO-1, and Na(+)/K(+)-ATPase alpha1, and could easily be marked with a fluorescent tracker. This study demonstrates the feasibility of reconstructing a highly cellular and healthy corneal endothelium on devitalized human corneal stromas. PMID- 19125644 TI - Macrophage participation in the degradation and remodeling of extracellular matrix scaffolds. AB - Biologic scaffolds composed of extracellular matrix (ECM) are widely used to facilitate remodeling and reconstruction of a variety of tissues in both preclinical animal studies and human clinical applications. The mechanisms by which such scaffolds influence the host tissue response are only partially understood, but it is logical that the mononuclear macrophage cell population plays a central role. The present study evaluated the role of macrophages that derive from peripheral blood in the degradation of ECM scaffolds. An established rat body wall reconstruction model was used to evaluate the degradation of carbodiimide (CDI)-crosslinked scaffolds composed of porcine small intestinal submucosa (SIS), noncrosslinked SIS, and autologous body wall. To assess the role of circulating macrophages in the degradation process, the degradation of each scaffold was assessed with and without macrophage depletion caused by administration of clodronate-containing liposomes. Results showed that peripheral blood monocytes are required for the early and rapid degradation of both SIS scaffolds and autologous body wall, and that CDI crosslinked SIS is resistant to macrophage-mediated degradation. PMID- 19125645 TI - Synthesis of a tissue-engineered periosteum with acellular dermal matrix and cultured mesenchymal stem cells. AB - Periosteal grafts can aid in bone repair by providing bone progenitor cells and acting as a barrier to scar tissue. Unfortunately, these grafts have many of the same disadvantages as bone grafts (donor site morbidity and limited donor sites). In this article, we describe a method of synthesizing a periosteum-like material using acellular human dermis and osteoblasts or mesenchymal stem cells (MSC). We show that osteoblasts readily attach to and proliferate on the acellular human dermis in vitro. In addition, osteoblasts retained the potential for differentiation in response to bone morphogenetic protein stimulation. Cells grown on the acellular human dermis were efficiently transfected with adenoviruses with no evidence of cellular toxicity. To assess for in vivo cell delivery and bone-forming potential, the acellular human dermis was seeded with green fluorescent protein (GFP)-positive MSCs, transfected with bone morphogenetic protein 2, wrapped around the adductor muscle in syngeneic mice, and used to treat critical-sized mandibular defects in nude rats. After 3 weeks, GFP-positive cells were still present, and bone had replaced the interface between the muscle and the constructs. After 6 weeks, critical-sized bone defects had been successfully healed. In conclusion, we show that an acellular human dermis can be used to synthesize a tissue-engineered periosteum capable of delivering cells and osteoinductive proteins. PMID- 19125646 TI - Self-assembling DNA quadruplex conjugated to MRI contrast agents. AB - We report the preparation and magnetic resonance (MR) characterization of new MRI contrast agents based on gadolinium complexes conjugated to a self-assembling DNA quadruplex scaffold. As a single gadolinium-DOTA chelated DNA strand, the r(1) molar relaxivity is 6.4 mM(-1) s(-1) per Gd and increases to 11.7 mM(-1) s(-1) per Gd upon formation of a DNA quadruplex. Similar results were obtained when a gadolinium-DOTA dendrimer was conjugated to DNA, with the r(1) molar relaxivity increasing to 12.9 mM(-1) s(-1) per Gd upon the formation of DNA quadruplex, compared to that of 6.0 mM(-1) s(-1) for a single strand of gadolinium-DOTA dendrimer chelate. This yields an r(1) molar relaxivity of 154.8 and 46.8 mM(-1) s(-1) per DNA quadruplex based on DOTA dendrimer or monomer, respectively. Importantly, the DNA quadruplex scaffold is approximately 2.5 nm(3) in size, potentially enabling this type of contrast agent to be used for targeted delivery in vivo to detect specific cells or tissues, even behind intact blood vessels. PMID- 19125648 TI - Detecting protein-ligand binding on supported bilayers by local pH modulation. AB - Herein, we describe a highly sensitive technique for detecting protein-ligand binding at the liquid/solid interface. The method is based upon modulation of the interfacial pH when the protein binds. This change is detected by ortho-Texas Red DHPE, which is doped into supported phospholipid bilayers and used as a pH sensitive dye. The dye molecule fluoresces strongly at acidic pH values but not basic ones and has an apparent pK(A) of 7.8 in 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine membranes containing 0.5 mol % biotin-cap-PE. This method was used to detect antibiotin/biotin binding interactions as well as the binding of cholera toxin B subunits to GM(1). Since these proteins are negatively charged under the conditions of the experiment the interface became slightly more acidic upon binding. In each case, the equilibrium dissociation constant was determined by following the rise in fluorescence as protein was introduced. This change is essentially linear with protein coverage under the conditions employed. For the biotin/antibiotin system it was determined that K(D) = 24 +/- 5 nM, which is in excellent agreement with classical measurements made by total internal reflection fluorescence microscopy involving fluorophore-conjugated antibody molecules. Moreover, the limit of detection was approximately 350 fM at the 99% confidence level. This corresponds to 1 part in 69,000 of the K(D) value. Such a finding compares favorably with surface plasmon resonance studies of similar systems and conditions. The assay could be run in imaging mode to obtain multiple simultaneous measurements using a CCD camera. PMID- 19125649 TI - Dual delivery of placental growth factor and vascular endothelial growth factor from poly(hydroxyethyl methacrylate-co-methyl methacrylate) microcapsules containing doubly transfected luciferase-expressing L929 cells. AB - Placental growth factor (PlGF) combined with vascular endothelial growth factor (VEGF(164)) did not result in greater luciferase expression of a microencapsulated genetically transfected mouse fibroblast cell line (L929 cells) implanted subcutaneously in nominally syngeneic C3H/HeJ mice than in a control without growth factors. The intent had been to increase the maturity status of VEGF-generated vessels in the implant site by co-delivery of PlGF and thereby to effect an improvement in the persistence of luciferase expression, a marker of encapsulated cell viability. L929 cells were doubly transfected with luciferase and PlGF or luciferase and VEGF. Two hundred microcapsules containing a 1:1 mixture of the two transfected cells were implanted minimally invasively in Matrigel on one side of the mouse; the other side contained 200 microcapsules containing cells expressing luciferase only. Luciferase expression, reflective of encapsulated cell number, peaked at day 21 in both groups at about three times the value at day 1. In contrast with the immature vessels produced with VEGF alone (as reported earlier), the vessels produced here were more mature at day 14, and there were more such vessels than in the control group, although by day 21, there was a mixture of mature and immature vessels with PlGF, consistent with the premise that the maturity status of new vessels is PlGF dose dependent. Furthermore, anti-L929 antibodies (according to flow cytometry) and CD3-positive cells (according to histology) were detected in mice receiving unencapsulated cells, suggesting that there may be minor MHC (major histocompatibility complex) alloantigens and an adaptive immune response in this animal model. Thus successful modulation of the host response to microencapsulated cells may require enhanced vascularization and manipulation of the immune response, even with nominally syngeneic cells. PMID- 19125647 TI - Radioimmunoimaging with longer-lived positron-emitting radionuclides: potentials and challenges. AB - Radioimmunoimaging and therapy has been an area of interest for several decades. Steady progress has been made toward clinical translation of radiolabeled monoclonal antibodies for diagnosis and treatment of diseases. Tremendous advances have been made in imaging technologies such as positron emission tomography (PET). However, these advances have so far eluded routine translation into clinical radioimmunoimaging applications due to the mismatch between the short half-lives of routinely used positron-emitting radionuclides such as (18)F versus the pharmacokinetics of most intact monoclonal antibodies of interest. The lack of suitable positron-emitting radionuclides that match the pharmacokinetics of intact antibodies has generated interest in exploring the use of longer-lived positron emitters that are more suitable for radioimmunoimaging and dosimetry applications with intact monoclonal antibodies. In this review, we examine the opportunities and challenges of radioimmunoimaging with select longer-lived positron-emitting radionuclides such as (124)I, (89)Zr, and (86)Y with respect to radionuclide production, ease of radiolabeling intact antibodies, imaging characteristics, radiation dosimetry, and clinical translation potential. PMID- 19125650 TI - Tissue-engineered small-caliber vascular graft based on a novel biodegradable composite fibrin-polylactide scaffold. AB - Small-caliber vascular grafts (< or =5 mm) constructed from synthetic materials for coronary bypass or peripheral vascular repair below the knee have poor patency rates, while autologous vessels may not be available for harvesting. The present study aimed to create a completely autologous small-caliber vascular graft by utilizing a bioabsorbable, macroporous poly(L/D)lactide 96/4 [P(L/D)LA 96/4] mesh as a support scaffold system combined with an autologous fibrin cell carrier material. A novel molding device was used to integrate a P(L/D)LA 96/4 mesh in the wall of a fibrin-based vascular graft, which was seeded with arterial smooth muscle cells (SMCs)/fibroblasts and subsequently lined with endothelial cells. The mold was connected to a bioreactor circuit for dynamic mechanical conditioning of the graft over a 21-day period. Graft cell phenotype, proliferation, extracellular matrix (ECM) content, and mechanical strength were analyzed. alpha-SMA-positive SMCs and fibroblasts deposited ECM proteins into the graft wall, with a significant increase in both cell number and collagen content over 21 days. A luminal endothelial cell lining was evidenced by vWf staining, while the grafts exhibited supraphysiological burst pressure (>460 mmHg) after dynamic cultivation. The results of our study demonstrated the successful production of an autologous, biodegradable small-caliber vascular graft in vitro, with remodeling capabilities and supraphysiological mechanical properties after 21 days in culture. The approach may be suitable for a variety of clinical applications, including coronary artery and peripheral artery bypass procedures. PMID- 19125651 TI - Access to poly-beta-peptides with functionalized side chains and end groups via controlled ring-opening polymerization of beta-lactams. AB - Poly-beta-peptides are attractive for biomedical applications because the backbone is similar enough to that of proteins for biocompatibility, but the backbone is sufficiently unnatural that these polymers evade proteolytic degradation. Prior investigations of poly-beta-peptides have been hindered by two principal limitations: (1) most known examples are insoluble, and (2) the range of accessible side chain functionality has been quite limited (mostly simple hydrocarbon units). The present study describes innovations in poly-beta-peptide synthesis that enable the preparation of diversely functionalized examples and provide the basis for broad exploration of the properties and applications of these nylon-3 materials. We describe several beta-lactams with a protected amino group in their side chain that readily undergo ring-opening polymerization (ROP). These monomers are available in large quantities via N-chlorosulfonylisocyanate (CSI) cycloaddition reactions with functionalized alkenes; previously CSI reactions have been limited to alkenes with hydrocarbon substituents. Postpolymerization deprotection of the amino groups leads to water-soluble poly beta-peptides. In addition, we introduce a simple co-initiation strategy that allows placement of a wide variety of functional groups at the N-termini of poly beta-peptide chains. ROP involving the new beta-lactams and co-initiation strategy exhibits characteristics of a controlled polymerization and enables the preparation of amphiphilic block copolymers. We have recently shown that cationic copoly-beta-peptides made available by these innovations mimic the selective antibacterial activity of host-defense peptides; the results described here provide the foundation for further exploration of this valuable activity and for the pursuit of other biological applications such as DNA/siRNA delivery and tissue engineering. PMID- 19125652 TI - Manipulating spheres that sink: assembly of micrometer sized glass spheres for optical coupling. AB - Previous work (Tull, E. J.; Bartlett, P. N.; Ryan, K. R. Langmuir 2007, 14, 7859; Tull, E. J.; Bartlett, P. N. Colloids Surf., A 2008, 327, 71) has demonstrated that, since convective forces have little influence over spheres in unstable sols, many conventional assembly techniques are ineffective for the organization of glassy spheres > or =5 microm in diameter into sparse patterns of interest for optoelectronic device applications. Carefully designed bifunctional substrates are required for both planar and "in cavity" assembly. In the present work, novel selective chemical modification of structured Ta2O5/glass substrates has been used to generate surfaces tailored to the assembly of glass microspheres from aqueous solution. Uniform arrays of 40-60 microm diameter glass spheres have been produced in pits of half-diameter depth, demonstrating the suitability of this technique for fabricating optical devices where light is coupled from a waveguide into the equator of a sphere. Individual aspects of the assembly process have been examined, and their influence over defect densities and the positioning of the assembled spheres is discussed. PMID- 19125653 TI - Molecular dynamics studies of the interactions of water and amino acid analogues with quartz surfaces. AB - The interactions of silica surfaces with water and biomolecules are of considerable significance in bio- and nanotechnology and in geochemistry. An important goal in the fields of biomineralization and biomimetics is to fine-tune these interactions for the control, e.g., of assembly of materials at the nanoscale. Here we report molecular dynamics simulations of fully hydroxylated alpha-quartz (1010), (0001), and (0111) surfaces in explicit water. We also present free energy estimates of adsorbing water and analogues of amino acid side chains onto the quartz (1010) surface. We find that at least two layers of structured water form on the surface, which is driven by the formation of a strong hydrogen bond network at the interface. Interestingly, we find that the free energy change to move methane (analogue of the side chain of alanine) from bulk water to the (1010) interface is favorable. We ascribe this to the presence of microscopic voids on the surface, which can accommodate small hydrophobic moieties and shield them from the solvent. These observations draw some useful insights into the possible mechanisms by which biomolecules, in particular peptides and proteins, bind to quartz and other silica surfaces. PMID- 19125654 TI - The impact of prostate size on perioperative outcomes in a large laparoscopic radical prostatectomy series. AB - PURPOSE: To clarify the effects of pathologic prostate specimen weight on perioperative outcomes in laparoscopic radical prostatectomy (LRP), a subject that has recently been analyzed in numerous smaller series. PATIENTS AND METHODS: Data from our Institution Review Board-approved database was queried with attention to operative, perioperative, and pathologic outcomes. For analysis, LRP patients were divided into three groups by pathologic specimen weight: <35 g, 35 to 70 g, and >70 g, and outcomes assessed. Outcomes were also analyzed using prostate weight as a continuous variable by multivariate regression. RESULTS: Between April 2001 and April 2007, 802 consecutive patients underwent LRP for localized prostate cancer, and complete perioperative data were available for 720 (90%) of these men. Mean age, body mass index (BMI), preoperative prostate specific antigen (PSA) and postoperative Gleason score were 57.6 years, 26.7 kg/m(2), 5.9 ng/mL, and 6.3, respectively. Mean specimen weight was 51.3 g. When compared with lighter counterparts, patients with the heaviest glands were older (P < 0.01), had a higher PSA level (P < 0.01), and had a higher percentage of pathologically organ-confined disease (P < 0.01). By multivariate regression analysis, increasing prostate weight was associated with longer operative times, more blood loss, longer lengths of stay, and more perioperative complications (all P < 0.05). Of note, smaller glands trended toward a higher rate of positive surgical margins overall (P = 0.07) and in pT(2) disease (P = 0.05), but there was no association between surgical margins and gland size in pT(3) disease (P = 0.27). Increasing BMI was also independently predictive of positive margins regardless of prostate size (P < 0.01). CONCLUSIONS: Although perioperative outcomes are generally excellent after LRP irrespective of gland size, a larger prostate size is associated with longer operative time, more blood loss, longer length of stay, and increased complications. Patients with smaller glands and organ-confined disease appear to have a higher rate of positive surgical margins. PMID- 19125656 TI - Ablation of bull prostate using novel bipolar radiofrequency ablation probe. AB - PURPOSE: We describe the use of a novel bipolar radiofrequency (RF) system for the destruction of prostate tissue in an ex vivo model. MATERIALS AND METHODS: A bipolar RF delivery system (Trod Medical, France) was linked to a 500 kHz generator. Eight lesions were created in an ex vivo bull prostate model using 7 mm (n = 4) and 10-mm probes (n = 4). Ablation was performed for 150 seconds. Temperature was recorded at the center of the lesion (Tc) and at the periphery (Tp) of the prostate. Tissue damage, both within and without the intended destruction zones, was assessed. The distance from Tc to Tp was recorded. RESULTS: All lesions created with either the 7-mm or 10-mm bipolar RF ablation (RFA) probes demonstrated complete tissue destruction only within the intended zone. Using the 7-mm probe, the mean Tc was 82 degrees C and the mean Tp 34 degrees C. The mean distance from the edge of the lesion to the periphery was 5 mm. Impedance was 62 Ohm, and power was 4 watts during ablation with the 7-mm probe. With the 10-mm probe, the mean Tc and Tp were 70 degrees C and 41 degrees C, respectively. The mean distance from the edge of the lesion to the periphery was 5 mm. Impedance was 78 Ohm, and power was 4.5 watts during ablation with the 10-mm probe. CONCLUSIONS: In an ex vivo model, bipolar RFA is capable of producing lesions with precise margins. Spread of heat is limited, evidenced histologically and by significant temperature drop off. This technology holds promise in the management of benign and malignant urologic diseases. PMID- 19125657 TI - Empirical scoring functions for advanced protein-ligand docking with PLANTS. AB - In this paper we present two empirical scoring functions, PLANTS(CHEMPLP) and PLANTS(PLP), designed for our docking algorithm PLANTS (Protein-Ligand ANT System), which is based on ant colony optimization (ACO). They are related, regarding their functional form, to parts of already published scoring functions and force fields. The parametrization procedure described here was able to identify several parameter settings showing an excellent performance for the task of pose prediction on two test sets comprising 298 complexes in total. Up to 87% of the complexes of the Astex diverse set and 77% of the CCDC/Astex clean listnc (noncovalently bound complexes of the clean list) could be reproduced with root mean-square deviations of less than 2 A with respect to the experimentally determined structures. A comparison with the state-of-the-art docking tool GOLD clearly shows that this is, especially for the druglike Astex diverse set, an improvement in pose prediction performance. Additionally, optimized parameter settings for the search algorithm were identified, which can be used to balance pose prediction reliability and search speed. PMID- 19125658 TI - Landscape genetics of raccoons (Procyon lotor) associated with ridges and valleys of Pennsylvania: implications for oral rabies vaccination programs. AB - Raccoons are the reservoir for the raccoon rabies virus variant in the United States. To combat this threat, oral rabies vaccination (ORV) programs are conducted in many eastern states. To aid in these efforts, the genetic structure of raccoons (Procyon lotor) was assessed in southwestern Pennsylvania to determine if select geographic features (i.e., ridges and valleys) serve as corridors or hindrances to raccoon gene flow (e.g., movement) and, therefore, rabies virus trafficking in this physiographic region. Raccoon DNA samples (n = 185) were collected from one ridge site and two adjacent valleys in southwestern Pennsylvania (Westmoreland, Cambria, Fayette, and Somerset counties). Raccoon genetic structure within and among these study sites was characterized at nine microsatellite loci. Results indicated that there was little population subdivision among any sites sampled. Furthermore, analyses using a model-based clustering approach indicated one essentially panmictic population was present among all the raccoons sampled over a reasonably broad geographic area (e.g., sites up to 36 km apart). However, a signature of isolation by distance was detected, suggesting that widths of ORV zones are critical for success. Combined, these data indicate that geographic features within this landscape influence raccoon gene flow only to a limited extent, suggesting that ridges of this physiographic system will not provide substantial long-term natural barriers to rabies virus trafficking. These results may be of value for future ORV efforts in Pennsylvania and other eastern states with similar landscapes. PMID- 19125659 TI - Role of communally nesting ardeid birds in the epidemiology of West Nile virus revisited. AB - Although herons and egrets in the family Ardeidae frequently have been associated with viruses in the Japanese encephalitis virus serocomplex, communal nesting colonies do not appear to be a focus of early season and rapid amplification of West Nile virus (WNV) in California. Evidence for repeated WNV infection was found by testing living and dead nestlings collected under trees with mixed species ardeid colonies nesting above in an oak grove near the University of California arboretum in Davis and in a Eucalyptus grove at a rural farmstead. However, mosquito infection rates at both nesting sites were low and positive pools did not occur earlier than at comparison sites within the City of Davis or at the Yolo Bypass wetlands managed for rice production and waterfowl habitat. Black-crowned night herons (Nycticorax nycticorax) were the most abundant and frequently infected ardeid species, indicating that WNV may be an important cause of mortality among nestlings of this species. PMID- 19125660 TI - Zoonotic Bartonella species in fleas collected on gray foxes (Urocyon cinereoargenteus). AB - Bartonella spp. are fastidious, gram-negative, rod-shaped bacteria and are usually vector-borne. However, the vector has not been definitively identified for many recently described species. In northern California, gray foxes (Urocyon cinereoargenteus) are infected with two zoonotic Bartonella species, B. rochalimae and B. vinsonii subsp. berkhoffii. Fleas (range 1-8 fleas per fox) were collected from 22 (41.5%) of 54 gray foxes from urban and backcountry zones near Hoopa, California. The flea species were determined, and DNA was individually extracted to establish the Bartonella species harbored by these fleas. Of the 108 fleas collected, 99 (92%) were identified as Pulex simulans. Overall, 39% (42/108) of the fleas were polymerase chain reaction (PCR)-positive for Bartonella, with B. rochalimae and B. vinsonii subsp. berkhoffii identified in 34 (81%) and 8 (19%) of the PCR-positive fleas, respectively. There was no difference between the prevalence of Bartonella spp. in P. simulans for the urban and backcountry zones. Fourteen (64%) of the 22 foxes were Bartonella bacteremic at one or more of the capture dates. In 10 instances, both the foxes and the fleas collected from them at the same blood collection were Bartonella-positive. B. rochalimae was the predominant species identified in both foxes and fleas. The competency of Pulex fleas as a vector of B. rochalimae has not been confirmed and will need to be demonstrated experimentally. Pulex spp. fleas readily feed on humans and may represent a source of human exposure to zoonotic species of Bartonella. PMID- 19125661 TI - Rickettsia felis in cat fleas in Taiwan. AB - We describe the first detection of Rickettsia felis in cat fleas (Ctenocephalides felis) in Taiwan. Natural infections of R. felis in cat fleas were isolated and confirmed using polymerase chain reaction (PCR) and an immunofluorescence assay. The infection rate in individual fleas and the minimum infection rate in pooled fleas detected by the PCR method were found to be 18.8% (13/69) and 8.2% (8/97), respectively. Partial sequences of the plasmid pRF, 17-kDa antigen, and outer membrane protein A genes obtained from the samples are identical to those of R. felis URRWXCal2. Serological studies confirmed R. felis infection in two stray cats, as demonstrated by the presence of serum IgG antibodies against R. felis with an immunofluorescence assay titer of 1:320. PMID- 19125662 TI - Prevalence of Rift Valley fever immunoglobulin G antibody in various occupational groups before the 2007 outbreak in Tanzania. AB - A cross-sectional sero survey of 199 apparently healthy persons from various occupations was carried out in Tanga, Tanzania in November 2004 to investigate exposure to Rift Valley fever (RVF) virus. Sera were tested for the presence of antibodies to RVF virus by the inhibition enzyme-linked immunosorbent assay (ELISA) for detecting immunoglobulin G (IgG). All reactive sera were further tested by the capture ELISA test and specific RVF immunoglobulin M (IgM) assay. Eight (4.0%) tested positive for IgG and none of the samples tested positive for IgM. Among the occupational groups examined, the seroprevalence was 7.3%, 1.5%, and 9.5% in the abattoir workers, livestock keepers, and others categories, respectively. Seropositivity was higher in men (5.3%) than women (1.5 %) and increased markedly in men aged between 20 to 40 years, with no significant differences among the age groups and sexes. The results indicate that a small proportion of people in Tanga municipality were exposed to RVF virus infection prior to 2007 disease outbreak in Tanzania. These findings need to be taken into consideration when future disease control programs are implemented. PMID- 19125663 TI - Uranyl-lanthanide heterometallic complexes with cucurbit[6]uril and perrhenate ligands. AB - The reaction of uranyl and lanthanide nitrates with cucurbit[6]uril (CB6) in the presence of perrhenic acid and under hydrothermal conditions yields the novel heterometallic uranyl-lanthanide molecular complexes [UO2Ln(CB6)(ReO4)2(NO3)(H2O)7](ReO4)2 (Ln = Sm, Eu, Gd, Lu). Both metal cations are bound to carbonyl groups of the same CB6 portal, one for UO2(2+) and two for Ln3+. The uranium atom is also bound to one monodentate perrhenate ion and three aquo ligands, while the lanthanide is bound to one monodentate perrhenate and one nitrate ions, and four aquo ligands. Not only are these complexes rare examples of ReO4(-) bonding to f element ions, but the perrhenate bound to Ln is included in the CB6 cavity, thus providing the first case of inclusion of a tetrahedral oxoanion in this macrocycle. PMID- 19125664 TI - Double cleavage of sp2 C-H and sp3 C-H bonds on one metal center: DMF-appended cyclometalated platinum(II) and -(IV) porphyrins. AB - DMF-appended cyclometalated platinum(II) and -(IV) porphyrins were synthesized via the double cleavage of sp(2) C-H and sp(3) C-H bonds on one platinum center. PMID- 19125665 TI - Si-based benzylic 1,4-rearrangement/cyclization reaction. AB - The trans-selective hydrosilylation of ynones (1) yields beta-silylated enones (2) that undergo a benzylic 1,4-rearrangement/cyclization reaction in the presence of base, yielding 2,5-dihydro-1,2-oxasiloles (3). PMID- 19125666 TI - An efficient formal synthesis of (-)-clavosolide a featuring a "mismatched" stereoselective oxocarbenium reduction. AB - The enantioselective formal synthesis of the polyketide marine natural product ( )-clavosolide A is presented. The construction of the beta-C-glycoside subunit is highlighted by a one-pot oxocarbenium cation formation/reduction sequence. Yamaguchi dimerization afforded the diolide aglycon in 18 steps (longest linear sequence). PMID- 19125667 TI - Resins with identical specifications are not identical. Identifying a useful solid-phase resin. AB - The quality of the most commonly used support for solid-phase syntheses, polystyrene resin cross-linked with 1% of divinylbenzene, differs considerably even among different lots of resin from the same source. Determination of the swelling capacity of resins before carrying out solid-phase syntheses represents a very simple means of nondestructive presynthetic resin characterization. PMID- 19125668 TI - Oligomerization and autocatalysis of NH2BH2 with ammonia-borane. AB - The reactivity of NH(2)BH(2) in the presence of ammonia-borane (AB) is investigated using ab initio CCSD(T) simulations to answer the following three questions: How do AB and NH(2)BH(2) react? How do aminoborane species oligomerize apart from catalytic centers? Can the formation of experimentally observed products, especially cyclic N(2)B(2)H(7)-NH(2)BH(3), be explained through the kinetics of NH(2)BH(2) oligomerization in the presence of AB? AB is shown to react with NH(2)BH(2) by the addition of NH(3)-BH(3) across the N=B double bond, generating linear NH(3)BH(2)NH(2)BH(3). This species decomposes by surmounting a reasonable barrier to produce two NH(2)BH(2) and H(2). The generation of additional NH(2)BH(2) from NH(2)BH(2) and AB provides a pathway for autocatalytic NH(2)BH(2) production. The important intermediates along the oligomerization pathway include cyclic (NH(2)BH(2))(2) and linear NH(3)BH(2)NH(2)BH(3), both of which have been observed experimentally. Calculations show cyclic N(2)B(2)H(7) NH(2)BH(3), an aminoborane analogue of ethylcyclobutane, to be the kinetically preferred stable intermediate resulting from oligomerization of free NH(2)BH(2) over its isomers, cyclic B(2)N(2)H(7)-BH(2)NH(3) and cyclotriborazane, cyclic (NH(2)BH(2))(3). Simulations show cyclotriborazane formation to be kinetically slower than cyclic B(2)N(2)H(7)-NH(2)BH(3) formation and imply that formation of the cyclic species cyclotriborazane and cyclopentaborazane may be catalyzed by binding of NH(2)BH(2) to a catalytic metal center. Routes that may lead to larger noncyclic oligomers are suggested to be kinetically competitive. The highly reactive N=B double bonds of NH(2)BH(2) are shown to be of central importance in understanding aminoborane oligomerization. PMID- 19125669 TI - Dimensionally modulated, single-crystalline LiMPO4 (M= Mn, Fe, Co, and Ni) with nano-thumblike shapes for high-power energy storage. AB - We demonstrate an efficient and rapid microwave irradiated solvothermal method to prepare nanostructured lithium metal phosphates LiMPO(4) (M = Mn, Fe, Co, and Ni) within a short reaction time (5-15 min) at temperatures as low as 300 degrees C without requiring any post annealing at elevated temperatures. The highly viscous, high-boiling tetraethyleneglycol used as the solvent not only provides a reducing atmosphere to prevent the oxidation of M(2+) to M(3+) but also inhibits the agglomeration of the nanoparticles formed. The enhanced reaction rates facilitated by the dielectric volumetric heating of the microwave absorbing reactants led to the formation of highly crystalline, phase-pure LiMPO(4) powders. The samples are characterized by X-ray diffraction, Raman spectroscopy, transmission electron microscopy (TEM), and electrochemical measurements in lithium cells. High-resolution TEM studies reveal the formation of single crystalline LiMPO(4) with nano-thumblike shapes. The dimensionally modulated nano thumblike shapes with the lithium diffusion direction (b axis) along the shorter dimension are particularly beneficial to achieve high-power capability in lithium ion cells. Subsequent networking of the single-crystalline LiMPO(4) nano-thumps with multiwalled carbon nanotubes by a simple solution-based mixing at ambient temperatures to overcome the electronic conductivity limitations offers excellent electrochemical performance in lithium ion cells. PMID- 19125670 TI - Synthesis of urchin-like VO2 nanostructures composed of radially aligned nanobelts and their disassembly. AB - Urchin-like VO(2)(B) nanostructures composed of radially aligned nanobelts have been synthesized by the homogeneous reduction reaction between peroxovanadic acid and oxalic acid under hydrothermal conditions. The influences of synthetic parameters, such as reaction times and the concentration of oxalic acid, on the morphologies and crystals of the resulting products have been investigated. The formation of VO(2)(B) nanostructures undergoes a reduction-dehydration phase transition and disassembly process. As the reaction time increases from 2 to 6 h, the diameters of urchin-like V(10)O(24) x 12 H(2)O nanostructures increase from 1 2 microm to 3-6 microm. After 12 h, urchin-like V(10)O(24) x 12 H(2)O nanostructures can be transformed in situ into VO(2)(B) nanostructures composed of radially aligned nanobelts, which can be described by a possible reduction dehydration phase-transition process. The urchin-like VO(2)(B) nanostructures are disassembled into dissociated VO(2)(B) nanobelts with the reaction time increased to 24 h. The shapes of the dissociated VO(2)(B) nanobelts can be controlled by adjusting the concentration of oxalic acid. PMID- 19125671 TI - Reduced susceptibility to vancomycin influences pathogenicity in Staphylococcus aureus infection. AB - In the present study, we demonstrated the utility of the nonmammalian model system Galleria mellonella for studying the pathogenesis of Staphylococcus aureus infection. By use of clinical and laboratory strains that had been exposed to vancomycin, we showed that both agr functional status and vancomycin minimum inhibitory concentration are determinants associated with the virulence of S. aureus in G. mellonella. These results show that G. mellonella can be effectively used to facilitate the in vivo study of S. aureus virulence and, more specifically, the relationship between antibiotic drug resistance and the pathogenesis of infection. PMID- 19125672 TI - In leishmaniasis due to Leishmania guyanensis infection, distinct intralesional interleukin-10 and Foxp3 mRNA expression are associated with unresponsiveness to treatment. AB - The presence of intralesional natural regulatory T cells, characterized by the expression of Foxp3 mRNA, was analyzed in patients with localized leishmaniasis due to Leishmania guyanensis infection that was unresponsive to treatment with pentamidine isethionate. Foxp3 mRNA levels were associated with unresponsiveness to treatment among patients with a lesion duration of 1 month, but this association was not observed among patients with a lesion duration of <1 month. In conclusion, high intralesional expression of Foxp3 might be an indicator of poor response to treatment, depending on the duration of lesions. PMID- 19125673 TI - Lactoferrin as a natural regimen for selective decontamination of the digestive tract: recombinant porcine lactoferrin expressed in the milk of transgenic mice protects neonates from pathogenic challenge in the gastrointestinal tract. AB - BACKGROUND: Nosocomial infection with antibiotic-resistant strains is a major threat to critical care medicine. Selective decontamination of the digestive tract (SDD) is one of the strategies used to reduce ventilator-associated pneumonia and sepsis in critically ill patients. In the present study, we performed pathogenic challenges of the digestive tract in a transgenic milk-fed animal model to test whether porcine lactoferrin (pLF) is an effective SDD regimen. METHODS: Transgenic mice expressing recombinant pLF in their milk at a mean+/-SD concentration of 120+/-13.6 mg/L during the lactation stage fed normal CD-1 mice pups for 4 weeks. The pups were subsequently challenged with pathogenic Escherichia coli, Staphylococcus aureus, and Candida albicans. RESULTS: Compared with the control groups fed wild-type (normal) milk, the groups fed pLF-enriched milk demonstrated statistically significant improvements in weight gain; lower bacterial numbers in intestinal fluid, blood, and liver; healthier microvilli in the small intestine; and alveoli in the lungs. CONCLUSIONS: Our results showed that oral administration of pLF-enriched milk to mice led to broad-spectrum antimicrobial activity in the digestive tract and protected the mucosa of the small intestine from injury, implying that pLF can be used as an effective SDD regimen. PMID- 19125674 TI - Controlled, household-randomized, open-label trial of the effect of treatment of Helicobacter pylori infection on iron deficiency among children in rural Alaska: results at 40 months. AB - BACKGROUND: Helicobacter pylori infection treatment was found not to reduce the prevalence of iron deficiency or anemia among Alaska Native children at 14 months after treatment initiation. We hypothesized that 14 months was to early to resolve H. pylori-induced gastric damage. Consequently, we conducted a 40-month follow-up. METHODS: We enrolled 219 children 7-11 years old who had H. pylori infection (as diagnosed by (13)C-labeled urea breath test) and iron deficiency (serum ferritin level, <22.47 pmol/L) in a controlled, household-randomized trial of the effect of treatment of H. pylori on iron deficiency and anemia (hemoglobin level, <115 g/L). At 40 months, 176 children were evaluated. RESULTS: Forty-four (52%) of 85 children in the intervention group and 53 (58%) of 91 in the control group had iron deficiency (adjusted relative risk [ARR], 0.92 [95% confidence interval {CI}, 0.68-1.26]), versus 4 (5%) and 17(19%), respectively, with both iron deficiency and anemia (ARR, 0.25 [95% CI, 0.09-0.73]). Reinfection occurred among 33 (52%) of 64 children who had cleared their infection. H. pylori-negative children had lower prevalences of iron deficiency (ARR, 0.62 [95% CI, 0.38-1.01]) and iron deficiency and anemia (ARR, 0.22 [95% CI, 0.03-1.50]), compared with H. pylori -positive children. CONCLUSIONS: The resolution of H. pylori infection for >14 months modestly reduced the prevalence of iron deficiency and substantially reduced the prevalence of iron deficiency and anemia. H. pylori likely plays a casual role in hematological outcomes for some children. PMID- 19125675 TI - Nontypeable Haemophilus influenzae adhesin protein E: characterization and biological activity. AB - The adhesin protein E (PE) of the human respiratory pathogen nontypeable Haemophilus influenzae (NTHi) exists in all clinical isolates. In the present study, NTHi adherence to epithelial cells of various origins was further analyzed. The number of intraepithelial PE-deficient NTHi was decreased compared with PE-expressing NTHi. Interestingly, PE-expressing NTHi or Escherichia coli transformants, in addition to soluble recombinant PE22-160 without a lipid moiety, induced a proinflammatory cell response. The adhesive PE domain was defined within PE84-108, and preincubation of epithelial cells with this peptide blocked adhesion of several clinical NTHi isolates. Mice immunized with PE84-108 cleared NTHi up to 8-fold more efficiently on pulmonary challenge than did mice immunized with a control peptide. Finally, anti-PE mouse antibodies from vaccinated mice prevented NTHi adhesion. Our data suggest that the ubiquitous adhesin PE plays an important role in the pathogenesis of NTHi infection. PMID- 19125677 TI - Hierarchical FeWO4 microcrystals: solvothermal synthesis and their photocatalytic and magnetic properties. AB - Highly hierarchical platelike FeWO(4) microcrystals have been synthesized by a simple solvothermal route using FeCl(3) x 6 H(2)O and Na(2)WO(4) x 2 H(2)O as precursors, where ethylene glycol (EG) plays an important role as a capping agent in directing growth and self-assembly of such unique structures. In addition, a certain amount of CH(3)COONa (NaAc) was necessary for the formation of such unique FeWO(4) microstructures. The photocatalytic property of as-synthesized hierarchical FeWO(4) microcrystals has been first studied, which shows excellent photocatalytic activity for the degradation of rhodamine B (RhB) under UV and visible light irradiation (modeling sunlight). Moreover, magnetic measurement indicates that hexangular FeWO(4) platelike microcrystals show a small ferromagnetic ordering at low temperature because of spin-canting of antiferromagnetic materials and surface spins of FeWO(4) nanoparticles. PMID- 19125676 TI - Gnotobiotic piglet infection model for evaluating the safe use of antibiotics against Escherichia coli O157:H7 infection. AB - BACKGROUND: Shiga toxin (Stx)-producing Escherichia coli (STEC), especially O157:H7, cause bloody diarrhea, and in 3%-15% of individuals the infection leads to hemolytic uremic syndrome (HUS) or other complications. Use of antibiotics to treat STEC infections is controversial. Here, we describe the use of piglets to evaluate the efficacy and mechanism of action of antibiotics in these infections. METHODS: The effects of 2 antibiotics on STEC toxin production and their mechanisms of action were first determined by enzyme-linked immunosorbent assay and subsequently evaluated clinically in the gnotobiotic piglet infection model. RESULTS: In vitro treatment of clinical and isogenic strains with ciprofloxacin increased the production of Stx2 via phage induction but not the production of Stx1. Azithromycin caused no significant increase in toxin production. After treatment with ciprofloxacin, infected piglets had diarrhea and the severe fatal neurological symptoms associated with Stx2 intoxication. Characteristic petechial hemorrhages in the cerebellum were more severe in ciprofloxacin-treated animals than in control animals. In contrast, azithromycin-treated piglets survived the infection and had little or no brain hemorrhaging. CONCLUSIONS: The increased in vitro toxin production caused by ciprofloxacin was strongly correlated with death and an increased rate of cerebellar hemorrhage, in contrast to the effect of azithromycin. The piglet is a suitable model for determining the effectiveness and safety of antibiotics available to treat patients. PMID- 19125678 TI - Importance of urinary tract infection to antibiotic use among hospitalized patients. AB - Many patients with asymptomatic bacteriuria receive extended courses of broad spectrum antibiotics. Antibiotic use was analyzed in patients admitted to the hospital with urinary tract infection. Strategies to optimize antibiotic use for such patients are discussed and include implementing a process whereby a urine culture is automatically performed if a urinalysis result suggests infection. PMID- 19125679 TI - Secular trends of hospitalization with vancomycin-resistant enterococcus infection in the United States, 2000-2006. AB - Although the incidence of hospitalizations with infection due to vancomycin resistant pathogens in the United States remained stable during 2000-2003, it increased from 4.60 to 9.48 hospitalizations per 100,000 population during 2003 2006. Hospitalizations with infection due to vancomycin-resistant pathogens also increased as a proportion of all US hospitalizations, from 3.16 to 6.51 hospitalizations with VRE infection per 10,000 total hospitalizations during 2003 2006. The number of hospitalizations with infection due to vancomycin-resistant pathogens is increasing in the United States. Because infection due to vancomycin resistant organisms is associated with poor outcomes, the epidemiology of this trend needs further exploration. PMID- 19125680 TI - Risk factors associated with surgical site infection after pediatric posterior spinal fusion procedure. AB - OBJECTIVE: To identify risk factors associated with surgical site infection (SSI) after pediatric posterior spinal fusion procedure by examining characteristics related to the patient, the surgical procedure, and tissue hypoxia. DESIGN: Retrospective case-control study nested in a hospital cohort study. SETTING: A 475-bed, tertiary care children's hospital. METHODS: All patients who underwent a spinal fusion procedure during the period from January 1995 through December 2006 were included. SSI cases were identified by means of prospective surveillance using National Nosocomial Infection Surveillance system definitions. Forty-four case patients who underwent a posterior spinal fusion procedure and developed an SSI were identified and evaluated. Each case patient was matched (on the basis of date of surgery, +/-3 months) to 3 control patients who underwent a posterior spinal fusion procedure but did not develop an SSI. Risk factors for SSI were evaluated by univariate analysis and multivariable conditional logistic regression. Odds ratios (ORs), with 95% confidence intervals (CIs) and P values, were calculated. RESULTS: From 1995 to 2006, the mean annual rate of SSI after posterior spinal fusion procedure was 4.4% (range, 1.1%-6.7%). Significant risk factors associated with SSI in the univariate analysis included the following: a body mass index (BMI) greater than the 95th percentile (OR, 3.5 [95% CI, 1.5 8.3]); antibiotic prophylaxis with clindamycin, compared with other antibiotics (OR, 3.5 [95% CI, 1.2-10.0]); inappropriately low dose of antibiotic (OR, 2.6 [95% CI, 1.0-6.6]); and a longer duration of hypothermia (ie, a core body temperature of less than 35.5 degrees C) during surgery (OR, 0.4 [95% CI, 0.2 0.9]). An American Society of Anesthesiologists (ASA) score of greater than 2, obesity (ie, a BMI greater than the 95th percentile), antibiotic prophylaxis with clindamycin, and hypothermia were statistically significant in the multivariable model. CONCLUSION: An ASA score greater than 2, obesity, and antibiotic prophylaxis with clindamycin were independent risk factors for SSI. Hypothermia during surgery appears to provide protection against SSI in this patient population. PMID- 19125681 TI - Multipronged intervention strategy to control an outbreak of Clostridium difficile infection (CDI) and its impact on the rates of CDI from 2002 to 2007. AB - OBJECTIVE: At the end of 2002, a new, more virulent strain of Clostridium difficile, designated BI/NAP1, was the cause of a massive outbreak of infection in the province of Quebec. This particular strain was associated with a dramatic increase in morbidity and mortality among affected patients in 2003-2004. We tested and implemented a multipronged infection control approach to curtail the rate of C. difficile infection (CDI). DESIGN: Five-year observational study. SETTING: A 554-bed, acute care tertiary hospital, the largest single medical center in Quebec, Canada. METHODS: To curtail the magnitude of the outbreak, we implemented a global strategy consisting of rapid C. difficile testing for all hospitalized patients who had at least 1 occurrence of liquid stool, the rapid isolation of patients infected with C. difficile in a dedicated ward with a specially trained housekeeping team, a global hand hygiene program, and the hiring of infection control practitioners. Antibiotic consumption at the institutional level was also monitored during the 5-year surveillance period. Cases of hospital-acquired CDI per 1,000 admissions were continuously monitored on a monthly basis during the entire surveillance period. RESULTS: The highest recorded CDI rate was 42 cases per 1,000 admissions in January 2004. Once additional infection control resources were put in place, the rate decreased significantly during the period from April 2005 to March 2007. During the 2003 2004 period, there were 762 cases of CDI (mean annual rate, 37.28 cases per 1,000 admissions) recorded in our study, compared with 292 cases of CDI (14.48 cases per 1,000 admissions) during the 2006-2007 period (OR, 0.379 [95% CI, 0.331 0.435]; p< .001), a 61% reduction. In March 2007, the equivalent of 4 full-time equivalent infection control practitioners were in place, which gave a ratio of 0.96 infection control practitioners per 133 beds in use, compared with the ratio of 0.24 infection control practitioners per 133 beds in use in 2003, and the total number of hours dedicated to cleaning and housekeeping increased by 26.2%. The total amount of antibiotics used in the hospital did not vary significantly from 2002 to 2007, although there were changes in the classes antibiotic used. CONCLUSION: The implementation of a multipronged intervention strategy to control the outbreak of CDI significantly improved the overall situation at the hospital and underlined the importance of investing in stringent infection control practices. PMID- 19125682 TI - GDNF plasma levels in spina bifida: correlation with severity of spinal damage and motor function. AB - Glial-derived neurotrophic factor (GDNF) is one of several powerful survival factors for spinal motoneurons that play a key role in sprouting, synaptic plasticity, and reorganization after spinal cord damage. The aim of this study was to investigate the expression of GDNF in plasma of children with spina bifida (SB) and to determine its correlation with both the severity of spinal cord damage and the motor function of these patients. To measure the GDNF expression, we collected plasma samples from 152 children with SB and in 149 matched controls. Endogenous GDNF levels were quantified using a two-site immuno enzymatic assay. The statistical analysis was performed using the Mann-Whitney two-tailed two-sample test. In children with SB the mean levels of GDNF (131.2 +/ 69.6 pg/mL) were significantly higher (p < 0.001) with respect to the mean levels of the control group (102.7 +/- 6.8 pg/mL). Moreover, in open SB, the GDNF levels (139.2 +/- 81.1 pg/mL) were significantly higher (p < 0.05) with respect to closed SB (117.2 +/- 41.3 pg/mL). In terms of the motor function of patients, we found that in children with poorer motor function, the GDNF levels (134.5 +/- 67.4 pg/mL) were higher, but not statistically significant (p < 0.1), than in patients with better motor outcome (122.3 +/- 72.2 pg/mL). Our study demonstrates GDNF over-expression in children with SB. This upregulation is significantly associated with the severity of spinal cord damage in SB patients and appears to correlate with poor motor function of children, representing an important biochemical marker of the severity of spine injury. PMID- 19125683 TI - Role of Nrf2 in protection against traumatic brain injury in mice. AB - Previous studies have shown that nuclear factor erythroid 2-related factor 2 (Nrf2) plays a unique role in many physiological stress processes. The present study investigated the role of Nrf2 in modulating traumatic brain injury (TBI) induced secondary brain injury. Wild-type Nrf2 (+/+) and Nrf2 (-/-)-deficient mice were subjected to a moderately severe weight-drop impact head injury. The absence of Nrf2 function in mice resulted in exacerbated brain injury as shown by the increased severity of neurological deficit, apoptosis, and brain edema at 24h after TBI. This exacerbation of brain injury in Nrf2-deficient mice was associated with increased mRNA and protein expression of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6), and with decreased mRNA expression and enzymatic activity of antioxidant and detoxifying enzymes including NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferase alpha-1 (GST-alpha1), compared with their wild-type counterparts after TBI. In combination, these results suggest that Nrf2 plays an important role in protecting TBI-induced secondary brain injury, possibly by regulating inflammatory cytokines and inducing antioxidant and detoxifying enzymes. PMID- 19125685 TI - Diversifying JAFC. PMID- 19125686 TI - Carotenoids in sea buckthorn ( Hippophae rhamnoides L.) berries during ripening and use of pheophytin a as a maturity marker. AB - Four cultivars of sea buckthorn berries were analyzed for their carotenoid and chlorophyll contents during ripening in three consecutive years. The different carotenoids generally increased in concentration during ripening and comprised from 120 to 1425 microg/g of DW of total carotenoids (1.5-18.5 mg/100 g of FW) depending on cultivar, harvest time, and year. GLM analyses revealed the effect of cultivar to be considerably larger than that of year and harvest time. The content of pheophytin a, a chlorophyll a derivate, steadily decreased during berry ripening. Pheophytin a therefore acted as a marker of the degree of ripening of sea buckthorn berries and was used here to convert harvest date into an estimated ripening time. PMID- 19125684 TI - Pathology dynamics predict spinal cord injury therapeutic success. AB - Secondary injury, the complex cascade of cellular events following spinal cord injury (SCI), is a major source of post-insult neuron death. Experimental work has focused on the details of individual factors or mechanisms that contribute to secondary injury, but little is known about the interactions among factors leading to the overall pathology dynamics that underlie its propagation. Prior hypotheses suggest that the pathology is dominated by interactions, with therapeutic success lying in combinations of neuroprotective treatments. In this study, we provide the first comprehensive, system-level characterization of the entire secondary injury process using a novel relational model methodology that aggregates the findings of approximately 250 experimental studies. Our quantitative examination of the overall pathology dynamics suggests that, while the pathology is initially dominated by "fire-like", rate-dependent interactions, it quickly switches to a "flood-like", accumulation-dependent process with contributing factors being largely independent. Our evaluation of approximately 20,000 potential single and combinatorial treatments indicates this flood-like pathology results in few highly influential factors at clinically realistic treatment time frames, with multi-factor treatments being merely additive rather than synergistic in reducing neuron death. Our findings give new fundamental insight into the understanding of the secondary injury pathology as a whole, provide direction for alternative therapeutic strategies, and suggest that ultimate success in treating SCI lies in the pursuit of pathology dynamics in addition to individually involved factors. PMID- 19125688 TI - When Wenzel and Cassie are right: reconciling local and global considerations. AB - The condition under which the Wenzel or Cassie equation correctly estimates the most stable contact angle is reiterated and demonstrated: these equations do hold when the drop size is sufficiently large compared with the wavelength of roughness or chemical heterogeneity. The numerical demonstrations somewhat mimic recent experiments that seemingly refuted the Wenzel and Cassie equations and show that these experiments were performed only for drops of sizes similar in order of magnitude to the wavelength of roughness or chemical heterogeneity. Under such conditions, the Wenzel and Cassie equations are a priori not expected to be valid. It is also explained that both the local equilibrium condition at the contact line and the global equilibrium condition involving the wetted area within the contact line are necessary and complementary. PMID- 19125687 TI - Cutaneous metabolic activation of carvoxime, a self-activating, skin-sensitizing prohapten. AB - Bioactivation of low molecular weight compounds in the skin can cause contact sensitization. We have previously shown that the alpha, beta-R-unsaturated oxime R-carvoxime [1, (R)-2-methyl-5-isopropenylcyclohex-2-enone oxime] is bioactivated to two diastereomeric highly reactive and strongly sensitizing alpha, beta-epoxy oxime metabolites. To investigate if this metabolic activation is catalyzed by the major cytochrome P450 (P450) enzymes found in human skin, incubations of 1 with a skinlike P450 cocktail in the presence of glutathione were carried out. We identified three glutathione conjugates in the incubation mixture arising from two diasteomeric alpha, beta-epoxy oxime metabolites of 1, thus showing that the metabolic activation of 1 is P450-mediated. A P450 identification study using the individual P450 enzymes present in the skinlike P450 cocktail showed the involvement of P450 1A1 and 1B1 and also to some extent 2B6. P450 1B1 metabolism of 1 was found to be stereoselective as glutathione conjugates from only one of the alpha, beta-epoxyoxime metabolites were identified (metabolite 2). Additionally, 1 was found to be an inducer of P450 1B1 (but not 1A1) in human monocyte-derived dendritic cells (moDCs) and to some extent in normal human epidermal keratinocytes. A further transcriptional gene expression change observed in moDCs was a 44-fold upregulation of IL-8, a marker often used for assessment of sensitizing potential of contact allergens. The autoinduction of P450 1B1 by 1 may be a key event in the development of contact allergy to 1 and may also explain why only metabolite 2, and not 3, was found to elicit an allergic response in mice sensitized to 1. Our data show that the alpha, beta unsaturated oxime 1 is bioactivated by human cutaneous P450, thus forming highly allergenic metabolites, and has the potential to induce its own bioactivation pathway, particularly in antigen-presenting cells. PMID- 19125689 TI - Redox-switched amphiphilic ionic liquid behavior in aqueous solution. AB - A new redox amphiphilic ionic liquid (AIL) containing ferrocene as a redox-active group was synthesized, 1-(11-ferrocenylundecyl)-3-methylimidazolium bromide (Fc11MIm+). Adsorption and aggregation of both reduced and oxidized forms of this ferrocenated AIL in aqueous solution were studied by surface tension measurements. The micellization was favored for the reduced ferrocenated AIL (Fc11MIm+) as compared with the oxidized ferrocenated AIL (Fc+11MIm+). Minimum areas at the air/aqueous solution interface were identical whereas limiting surface tensions were slightly different. This corroborated the formation of an expanded monolayer of redox active AIL at the interface. The electrochemical behavior of redox active AIL was investigated. The electrochemical responses of Fc11MIm+ aqueous solution interestingly differed, depending on its concentration. Below the cmc, the electrochemical reaction was dominated by ferrocenated AIL adsorbed onto the electrode surface; then above the cmc, it was controlled by the Fc11MIm+ diffusing to the electrode. For the latter, the electrochemical mechanism was suggested to couple with the disruption reaction of the reduced form micelles. PMID- 19125691 TI - Highly diastereoselective anti-aldol reactions of glycolate titanium enolates. AB - Diastereoselective aldol reactions of enolates of alpha-hydroxy ester with different aldehydes are metal tunable, providing either anti- or syn- products. The highest anti-selectivities were obtained with TiCl(OiPr)(3), while Ti(OiPr)(4) provides the opposite syn-products. PMID- 19125692 TI - Chondroitin 4-O-sulfotransferase-1 is required for somitic muscle development and motor axon guidance in zebrafish. AB - CS (chondroitin sulfate) has been implicated in a variety of biological processes during development. Its biological functions are closely associated with characteristic sulfated structures. Here, we report the characterization of a zebrafish counterpart of C4ST-1 (chondroitin 4-O-sulfotransferase-1) and its functional importance in embryogenesis. Recombinant C4ST-1 showed a substrate preference for chondroitin and catalysed the 4-O-sulfation of GalNAc residues, a highly frequent modification of CS in the embryos of zebrafish as well as other vertebrates. Whole-mount in situ hybridization revealed that C4ST-1 showed a distinct spatiotemporal expression pattern in the developing zebrafish embryo. During the segmentation stages, strong expression was observed along the body axis including the notochord and somites. Functional knockdown of C4ST-1 with specific antisense morpholino-oligonucleotides led to a marked decrease in the 4 O-sulfation and amount of CS in the embryos. Consistent with the preferential expression in the rostrocaudal axis, C4ST-1 morphants displayed morphological defects exemplified by a ventrally bent trunk and a curled and/or kinky tail, largely due to misregulated myotomal myod expression, implying perturbation of axial muscle differentiation in somites. Furthermore, the aberrant projection of spinal motor axons, which extended ventrally at the interface between the notochord and individual somites, was also observed in C4ST-1 morphants. These results suggest that 4-O-sulfated CS formed by C4ST-1 is essential for somitic muscle differentiation and motor axon guidance in zebrafish development. PMID- 19125694 TI - Nitration of cathepsin D enhances its proteolytic activity during mammary gland remodelling after lactation. AB - Proteomic studies in the mammary gland of control lactating and weaned rats have shown that there is an increased pattern of nitrated proteins during weaning when compared with controls. Here we report the novel finding that cathepsin D is nitrated during weaning. The expression and protein levels of this enzyme are increased after 8 h of litter removal and this up-regulation declines 5 days after weaning. However, there is a marked delay in cathepsin D activity since it does not increase until 2 days post-weaning and remains high thereafter. In order to find out whether nitration of cathepsin D regulates its activity, iNOS (inducible nitric oxide synthase)(-/-) mice were used. The expression and protein levels of this enzyme were similar to WT (wild-type) animals, but the proteolytic activity was significantly reduced during weaning in knockout compared to WT mice. in vitro treatment of recombinant human cathepsin D or lactating mammary gland homogenates with relatively low concentrations of peroxynitrite enhances the nitration as well as specific activity of this enzyme. Using MS, it has been shown that the residue Tyr168 was nitrated. All of these results show that protein nitration during weaning might be a signalling pathway involved in mammary gland remodelling. PMID- 19125693 TI - Human calcium/calmodulin-dependent serine protein kinase regulates the expression of p21 via the E2A transcription factor. AB - CASK (calcium/calmodulin-dependent serine protein kinase) is a kind of scaffolding protein that recruits or organizes other proteins at the plasma membrane to co-ordinate signal transduction pathways within the cytoplasm and nucleus. We have previously found that hCASK (human CASK) binds Id1 (inhibitor of DNA binding 1) through hCASK's GUK (guanylate kinase) domain and inhibits cell growth, probably via interactions with Id1. Overexpression of hCASK resulted in a reduced rate of cell growth, although inhibition of CASK via RNAi (RNA interference) promoted cell proliferation in ECV304 cells. This study revealed that hCASK regulates the protein and mRNA level of p21(wafi/cip1) (referred to throughout as p21), and activated the expression of p21 in a time-dependent manner. Two E-boxes in the proximal region at the TSS (transcription start site) play key roles in regulating hCASK-mediated p21 expression. We suggest that E2A (E12 and E47), a representative of the E proteins that binds the E-box elements, is a participant in the mediation of p21 expression by hCASK. The results of the present study suggest that hCASK regulation of cell growth might involve p21 expression, and that the bHLH (basic helix-loop-helix) transcription factor E2A probably participates in hCASK regulation of p21 expression. From these findings, we propose a novel proliferation signalling pathway mediated by hCASK. PMID- 19125695 TI - The inositol phosphatase MTMR4 is a novel target of the ubiquitin ligase Nedd4. AB - The inositol phosphatase, MTMR4 (myotubularin-related protein 4), was identified as a novel interactor of the ubiquitin ligase Nedd4 (neural-precursor-cell expressed developmentally down-regulated 4). hMTMR4 (human MTMR4) and Nedd4 co immunoprecipitated and co-localized to late endosomes. The PY (Pro-Tyr) motif of hMTMR4 binds to WW (Trp-Trp) domains of hNedd4. MTMR4 expression was decreased in atrophying muscle, whereas Nedd4 expression was increased and hMTMR4 was ubiquitinated by hNedd4, suggesting that this novel interaction may underlie the biological process of muscle breakdown. PMID- 19125697 TI - Effect of ischaemic exercise training of a normal limb on angiogenesis of a pathological ischaemic limb in rabbits. AB - The present study was designed to test the hypothesis that local exercise of a normal limb can promote angiogenesis in a pathological ischaemic limb. New Zealand White rabbits underwent left femoral artery ligation (Lig) and electrode implantation on the right sciatic nerve. The rabbits were randomly assigned to four groups: (i) Lig-N group, which did not receive ES (electrical stimulation); (ii) Lig-High group, which received high-intensity ES (2.5 mA, 40Hz for 1 ms) on the right hindlimb; (iii) Lig-Low group, which received low-intensity ES (0.3 mA, 40Hz for 1 ms) on the right hindlimb; (iv) Double-Lig-High group, which underwent femoral artery ligation on both hindlimbs and received high-intensity ES (2.5 mA, 40Hz for 1 ms) on the right hindlimb. The ES procedure included 5 min of stimulation, followed by 5 min of rest, and was repeated eight times a day for 4 weeks. Collateral circulation was examined grossly by angiography, resting blood flow was measured using the microspheres technique, and capillary supply was evaluated by immunohistochemistry. VEGF (vascular endothelial growth factor) mRNA and protein were analysed by real-time RT (reverse transcription)-PCR and Western blotting respectively. Collateral blood flow in all of the major muscles of the left hindlimb in the Lig-High group was highest among the four groups (P<0.01). Capillary supply (P<0.001), VEGF mRNA (P<0.01) and VEGF protein (P<0.01) in the gastrocnemius muscle increased remarkably in the Lig-High group; no statistically significant difference was observed among the other three groups. In conclusion, angiogenesis associated with an up-regulation of VEGF expression in pathological ischaemic limb may be facilitated by 4 weeks of physiological ischaemic exercise training in a normal limb. PMID- 19125696 TI - Functional analysis reveals effects of tobacco alternative oxidase gene (NtAOX1a) on regulation of defence responses against abiotic and biotic stresses. AB - Mitochondrial AOX (alternative oxidase) is the terminal oxidase of the CN (cyanide)-resistant alternative respiratory pathway in plants. To investigate the role of the tobacco AOX gene (NtAOX1a) (where Nt is Nicotiana tabacum) under deleterious conditions which could induce ROS (reactive oxygen species) accumulation, we generated and characterized a number of independent transgenic tobacco (N. tabacum) lines with altered NtAOX1a gene expression and AP (alternative pathway) capacity. AOX efficiently inhibited the production of low temperature-induced H2O2 and might be a major enzyme for scavenging H2O2 at low temperature. Furthermore, NtAOX1a may act as a regulator of KCN-induced resistance to TMV (tobacco mosaic virus) through the regulation of H2O2. Notably, a moderate accumulation of H2O2 under the control of NtAOX1a was crucial in viral resistance. Analysis of seed germination indicated an important role for NtAOX1a in germination under H2O2-induced oxidative stress when the CP (cytochrome pathway) was inhibited. These results demonstrate that NtAOX1a is necessary for plants to survive low temperature, pathogen attack and oxidative stress by scavenging ROS under these adverse conditions when the CP is restricted. PMID- 19125698 TI - Those are usually the sole proclamations for what is ultimately possible in each individual country that adopts defined and agreed upon joint principles. PMID- 19125699 TI - Collateral function in patients with coronary occlusion evaluated by 201 thallium scintigraphy. AB - The present study evaluated the impact of the angiographically documented collaterals on regional myocardial perfusion measured by 201thallium scintigraphy in patients with a chronic total occlusion. The study included 60 patients with chronic total occlusion who underwent rest-stress myocardial perfusion scintigraphy and coronary angiography. All patients had angiographic evidence of coronary collaterals. Patients were divided into two groups: group one had well developed coronary collateral vessels (n=35) and group II had poor coronary collateral development (n=25). Patients with chronic total occlusion had severe and extensive stress-induced myocardial perfusion defects regardless of the grade of angiographic coronary collaterals. The perfusion defects in the group with good collaterals were predominantly reversible, suggesting that coronary collaterals preserved myocardial viability in the regions subtended by a total coronary occlusion. A significant correlation between good collaterals with complete protection and poor collaterals with no protection was noted. Our results demonstrate a protective effect of collaterals on myocardial perfusion during coronary occlusion. The effective angiographic collaterals may prevent resting regional wall motion abnormalities but do not appear to protect against stress-induced perfusion defect. PMID- 19125700 TI - Effects of basal insulin analog and metformin on glycaemia control and weight as risk factors for endothelial dysfunction. AB - Obese patients with type 2 diabetes and impaired glucose tolerance are at increased risk of development of cardiovascular diseases. Endothelial dysfunction may be a reason for development of atherosclerosis and cardiovascular diseases. Lifestyle modification, increased physical activity, weight reduction, energy restricted diet and good glycaemia control can be useful for the endothelial function improvement and may decrease the risk of cardiovascular diseases. The aim of this study was to evaluate the effects of basal insulin analog and metformin on glycaemia control and weight as risk factors of endothelial dysfunction. Total of 15 patients (9 male and 6 female) with type 2 diabetes were studied. The patients were monitored over six months period. Glycated hemoglobin (HbA1c), fasting plasma glucose (FPG), postprandial plasma glucose (PPG), and body mass index (BMI) were observed. Mean age of the subjects was 53,4 +/- 6,27 years. Mean diabetes duration was 3,71 +/- 1,89 years. At the end of the study mean body mass index decreased from 27,5 +/- 1,45 kg/m2 to 25,7 +/-1,22 kg/m2. In this study we included diabetic patients with fasting glycaemia over 7 mmol/dm3, postmeal glycaemia over 7,8 mmol/dm3 and glycated hemoglobin over 7%. Prior to the study, the patients were treated with premix insulin divided in two daily doses and metformin after the lunch, which did not result in sufficient regulation of glycaemia. We started treatment with one daily insulin basal analog and three daily doses of metformin and monitored the above mentioned parameters. We advised patients to change their lifestyle, to practice energy restricted diet and to increase their daily physical activity. Insulin doses were titrated separately for each patient (0,7-1 IU/kg). Weight reduction was recorded after the study. Mean fasting glycaemia decreased from 8,6+/-0,49 mmol/dm3 to 7,04+/ 0,19 mmol/dm3 (p < 0,05). Mean postmeal glycaemia decreased from 9,74 +/- 0,79 mmol/dm3 to 7,6 +/- 0,43 mmol/dm3 (p<0,05). Mean HbA1c decreased from 8,80 +/- 0,59 % to 7,11 +/- 0,22 % (p<0,05). Treatment with one daily doses of basal insulin analog and three daily doses of metformin with lifestyle modification and weight reduction, in obese patients with type 2 diabetes can be useful for the endothelial function improvement and may decrease the risk of cardiovascular diseases. PMID- 19125701 TI - Microsatellite instability and loss of heterozygosity of tumor suppressor genes in Bosnian patients with sporadic colorectal cancer. AB - Considering its frequency, high mortality rate as well as many etiological mysteries colorectal cancer is a challenge to contemporary science. In our study we analyzed RER + and RER--phenotypes and their relations with clinical pathological characteristics of sporadic colorectal cancers. We also analyzed genetic alterations of tumor suppressor genes as well as their relation with microsatellite instability. The study was based on 54 tumor samples and 54 samples of the surrounding healthy tissue of patients with colorectal cancer. According to Amsterdam Criteria and Bethesda Criteria 35/54 or 64,81% belonged in the group of sporadic colorectal cancer. Mononucleotide marker Bat 25 showed instability in 48,57%; Bat 26 in 45,71% and Bat 40 in 29/35 82,86% of tumor samples. Considering dinucleotide markers, TP 53 showed instability in 54,29% and DS123 in 37,14% of tumor samples. Genetic alterations in tumor suppressor genes were found in tumor tissue: NM 23 in 54,29% samples, p53 in 51,43%, APC in 51,43%, DCC2 in 34,29%, RB1 in 22, 86% and DCC 1 in 28,57%. Our studies confirmed that genetic instability had an important role in the development of tumor type. Our results showed that mononucleotide marker Bat 40 might be used for an easy and fast screening procedure in Bosnian population, because it exhibited high percent of microsatellite instability and was in relation with RER+ phenotype. This investigation showed that different genetic alterations may occur during cancer development in each individual patient's tumor. These changes result in MMR inactivation, which causes RER+ phenotype. Our results suggest a connection between alteration in some tumor suppressor genes and MSI phenotype of sporadic colorectal cancer in Bosnian population. PMID- 19125702 TI - Bacterial vaginosis presence in sexually active women in Tuzla Canton area. AB - The goal of our research was to determine the presence of bacterial vaginosis in sexually active women in Tuzla Canton area. Diagnosis determination for bacterial vaginosis was conducted on the basis of three out of four internationally accepted criteria according to Amsel and isolation and identification of Gardnerella vaginalis (G. vaginalis) by standard microbiological procedures. Bacterial vaginosis was diagnosed in 20,5 % (41/200) women who asked for gynaecologist's help due to their personal discomfort, since significantly higher percentage of diagnosed bacterial vaginosis of 48,80% (41/84) was determined in women with personal discomfort typical for this disease. All relevant factors, according to available literature, for genesis of bacterial vaginosis were processed in this research. In respect to the obtained outputs, bacterial vaginosis is significantly more frequent occurrence in women who are not married, since the number of sexual partners, the time of the first sexual intercourse, the use of intrauterine contraceptive device and smoking do not cause the genesis of bacterial vaginosis. According to Nugent, an increased vaginal discharge with unpleasant odour after sexual discourse, its pH>4,5, a positive amino odour test, an occurrence of clue cells in a direct microscopic concoction of vaginal discharge and assessment of the state of vaginal flora for bacterial vaginosis are significantly more frequent occurrences in women with individual discomforts. It was proved that G. vaginalis is a dominant micro organism in 95% of women with clinical signs of vaginosis although it was isolated from vaginal discharge in 40 to 50% of healthy women. In our research, G. vaginalis was isolated in 63,41% of examined women with all signs of bacterial vaginosis, in 36,59% of examined women with one or more clinical signs of bacterial vaginosis and in 2,58% of examined women of control group without clinical signs. PMID- 19125703 TI - Spirulina platensis protects against renal injury in rats with gentamicin-induced acute tubular necrosis. AB - The present study was carried out to evaluate the renoprotective antioxidant effect of Spirulina platensis on gentamicin-induced acute tubular necrosis in rats. Albino-Wistar rats, (9male and 9 female), weighing approximately 250 g, were used for this study. Rats were randomly assigned to three equal groups. Control group received 0,9 % sodium chloride intraperitoneally for 7 days at the same volume as gentamicin group. Gentamicin group was treated intraperitoneally with gentamicin, 80 mg/kg daily for 7 days. Gentamicin+spirulina group received Spirulina platensis 1000 mg/kg orally 2 days before and 7 days concurrently with gentamicin (80 mg/kg i.p.). Nephrotoxicity was assessed by measuring plasma nitrite concentration, stabile metabolic product of nitric oxide with oxygen. Plasma nitrite concentration was determined by colorimetric method using Griess reaction. For histological analysis kidney specimens were stained with hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) stain. Plasma nitrite concentration and the level of kidney damage were significantly higher in gentamicin group in comparison both to the control and gentamicin+spirulina group. Spirulina platensis significantly lowered the plasma nitrite level and attenuated histomorphological changes related to renal injury caused by gentamicin. Thus, the results from present study suggest that Spirulina platensis has renoprotective potential in gentamicin-induced acute tubular necrosis possibly due to its antioxidant properties. PMID- 19125704 TI - Alcohol sclerotherapy in the treatment of symptomatic simple renal cysts. AB - The aim of this study was to evaluate the results of percutaneous drainage with single-session alcohol sclerotherapy in the treatment of symptomatic simple renal cysts. Thirty patients (16 men and 14 women, average age 54,7+/-11,3 years) with simple renal cysts were treated by ultrasound guided percutaneous aspiration and single-session alcohol sclerotherapy. Patient demographics, clinical characteristics, treatment outcome and complications were analyzed. Average reduction of cyst volume was 91,2%. Complete and partial resolution occurred in 10 (33%) and 14 (47%) cysts, respectively. Six cases were defined as failure with reduction of cyst volume <80%. Flank pain improved in all patients, regardless of complete or partial resolution. Minor complications occurred in 8 patients. The hospital stay was one day for all patients. Treatment of simple renal cysts is indicated when the cysts are sufficiently large and cause complaints or when associated with complications. Percutaneous treatment with alcohol sclerotherapy of benign renal cysts can be performed safely and effectively. PMID- 19125705 TI - The role of alpha-lipoic acid in diabetic polyneuropathy treatment. AB - Diabetic neuropathy represents late diabetes complications, and diabetes duration and long-term hyperglycemia are the main reasons for polyneuropathy. The goal was to estimate the effects of alpha-lipoic acid on symptoms of diabetic neuropathy after 600 mg i.v. for 3 weeks and 3 months of 300-600 mg of alpha-lipoic acid per os. This study has been designed as a multicentric, in 5-centers in B&H, carried out by 5 physicians with 20 diabetic patients each. Following parameters were monitored in 100 diabetics suffering from Type 1 and Type 2 diabetes, both men and women: diabetes duration, diabetes therapy, duration of polyneuropathy symptoms, height, weight, BMI (body mass index), subjective assessment of patients, objective examinations of physicians and subjective assessment of physicians. 100 diabetics, average age 61,36; oldest 79, youngest 40, suffered from diabetes in average 11,9 years. There were 35 men and 65 women, 16 with Type 1 and 80 with Type 2 diabetes, while 4 patients were not classified. 69 were having insulin therapy and 31 oral hypoglicemics. Shortest diabetic status was less than a year, and longest was 28 years. Average duration of polyneuropathic symptoms was 3,02 years, shortest was less than a year, and the longest was 15 years. Average height was 1,70 m, average weight 76,13 kg, and average BMI 26,51 kg/m2. Significant statistic differences in improvement were recorded (P>0,05) according to Fridman's test for repeated measurements compared to initial findings in assessments: sensory symptoms of polyneuropathy, pain sensations as polyneuropathy symptoms, total score of polyneuropathy symptoms, subjective assessment of patients, subjective findings of physicians, and significant differences were not find (P>0,05) in autonomous and motoric neuropathy. Based on the conducted study, we have concluded that the application of alpha-lipoic acid during 3 months has helped to decrease the symptoms of diabetic neuropathy and in only one case out of 100 included patients there was no subjective improvement after drug application. PMID- 19125706 TI - Correlation between refraction level and retinal breaks in myopic eye. AB - In this study we analyzed 180 myopic eyes in order to determine the refraction that is "critical" for the occurrence of retinal breaks as a main cause of retinal detachment. Detachment of retina involves separation of the neurosensory retina from the pigmented epithelium with the severe impairment of vision. After the focused ophthalmological assessment, determination of objective refraction, indirect binocular ophthalmoscopy, we compared the diagnosed retinal breaks according to the shape with the refraction. All the examined eyes were divided into six groups according to the axis length. Mean age of our patients was between 48,43 and 51,60 years with SD ranging from 13,88 to 18,45. The age differences among the groups were not statistically significant. This study included 102 (56,7%) male and 78 (43,3%) female patients and no statistically significant differences between genders was found regarding the occurrence of retinal breaks compared to refraction. The most dominant ruptures were the round ones (28,2%), followed by oval (25%), the category of multiple small holes (19,2%), horseshoe shaped (15,3%), and finally holes with operculum. In cases with myopic refraction ranging between 3,50 and 7,49 dsph, the frequency of retinal breaks statistically significantly differs from all other analyzed refractions. Also, there is positive correlation between the above mentioned myopic refraction and the frequency of retinal breaks. In order to prevent retinal detachment in a myopic eye, we suggest further thorough examinations of the eye fundus in patients with the above mentioned myopia. Diagnosing retinal breaks involves the application of adequate therapy: laser photocoagulation, cryotherapy, sclera buckling and pneumatic retinopexy. PMID- 19125707 TI - RT-PCR detection of HIV in Republic of Macedonia. AB - The aim of the study was to detect HIV RNA in seropositive patients using RT-PCR method and thus, to establish PCR methodology in the routine laboratory works. The total of 33 examined persons were divided in two groups: 1) 13 persons seropositive for HIV; and 2) 20 healthy persons - randomly selected blood donors that made the case control group. The subjects age was between 25 and 52 years (average 38,5). ELFA test for combined detection of HIV p24 antigen and anti HIV 1+2 IgG and ELISA test for detection of antibodies against HIV-1 and HIV-2, were performed for each examined person. RNA from the whole blood was extracted using a commercial kit based on salt precipitation. Detection of HIV RNA was performed using RT-PCR kit. Following nested PCR, the product was separated by electrophoresis in 1,5 % agarose gel. The result was scored positive if the band of 210bp was visible regardless of intensity. Measures of precaution were taken during all the steps of the work and HIV infected materials were disposed of accordingly. In the group of blood donors ELFA, ELISA and RT-PCR were negative. Assuming that prevalence of HIV infection is zero, the clinical specificity of RT PCR is 100 %. The analytical specificity of RT-PCR method was tested against Hepatitis C and B, Human Papiloma Virus, Cytomegalovirus, Herpes Simplex Virus, Rubella Virus, Mycobacterium tuberculosis, Chlamydia trachomatis. None of these templates yielded amplicon. In the group of 13 seropositive persons, 33 samples were analyzed. HIV RNA was detected in 15 samples. ELISA and ELFA test were positive in all samples. Different aliquots of the samples were tested independently and showed the same results. After different periods of storing the RNA samples at -70 masculineC, RT-PCR reaction was identical to the one performed initially. The obtained amplicons were maintained frozen at -20 masculineC for a week and the subsequently performed electrophoresis was identical to the previous one. The reaction is fast, simple for manipulation; with low detection level of 60 IU/ml. RT-PCR needs a small amount of RNA, as well as a small volume of sample. HIV RNA was detected in different periods of time with different clinical presentations in patients, with or without antiretroviral therapy. RT-PCR method gives the opportunity for reliable determination of HIV-1 RNA with border of detection of 60 IU/ml. The test is reproducible and has high analytical and clinical specificity. PMID- 19125708 TI - Congenital malformations of the central nervous system: clinical approach. AB - Central nervous system (CNS) malformations represent important factor of morbidity and mortality in children. The aim of the study was to determine the incidence, type and clinical features of CNS malformations in children who were admitted at the Neonatal and Child Neurology Department, Neonatal Intensive Care Unit and Paediatric Intensive Care Unit of Paediatric Clinic, University of Sarajevo Clinics Centre, from January 1st, 2002 to December 31st, 2006. There were total of 16520 admissions at the Paediatric Clinic over the studied period. CNS malformations, solitary or multiple, have been diagnosed in 100 patients (0,61%). The total number of various CNS malformations was 127. Lethal outcome was established in 9/100 cases (9%). The most frequent CNS malformations were neural tube defects 49/127 (38,6%). Hydrocephalus was seen in 34/127 (26,8%), microcephaly in 24/127 (18,9%), agenesis of corpus callosum in 10/127 (7,9%), Dandy Walker malformation in 6/127 (4,7%) and other CNS malformations in 4/127 (3,1%). In 20/100 of patients neural tube defect was associated with hydrocephalus (20%). CNS malformations were prenatally diagnosed in 13/100 of patients (13%). Primary prevention of CNS malformations can be improved in our country by better implementation of preconceptional folic acid therapy for all women of childbearing age. Secondary prevention by prenatal diagnosis requires advanced technical equipment and adequate education of physicians in the field of foetal ultrasonography. In our circumstances, prenatal diagnostics of CNS malformations is still not developed enough. PMID- 19125709 TI - Malignant pleural mesothelioma treated in Clinic for Pulmonary Diseases and Tuberculosis "Podhrastovi" in ten-year period (from 1998 to 2007). AB - Malignant pleural mesothelioma (MPM) is the most common primary malign tumour of pleura. The aim of this study was to evaluate cases of MPM diagnosed and treated in Clinic for Pulmonary Diseases and Tuberculosis "Podhrastovi" during ten-year period (1998-2007). Study is retrospective. The patients were analysed according to age, sex, histopathologic type of the tumour, cantonal distribution in Federation of Bosnia and Herzegovina and regimen of treatment. MPM presented 0,72% (0-1,56% per year) of all hospitalised malignant patients, and the greatest number of registered cases was in the year of 2007. The series included 16 male (57,14%) and 12 female (42,86%). Cases over 64 years old were the most frequent (14-50%) than 45-54 years (7- not 25%). Histopathology types of hospitalised cases of MPM: epitheloid form (8- 28,57%); sarcomatoid form (2- 7,14 %); other forms (18-64,29%). The most patients came from Canton Sarajevo (12-42,86%); ZE-DO canton (8-28,57%) and the UNA-SANA canton (5-17,86%). The therapy applied: chemotherapy (11-39,29%); radiotherapy (3-10,71%); chemotherapy + radiotherapy (4 14,29%); symptomatic therapy (10-35,71 %). PMID- 19125710 TI - Femoro-popliteal bypass above knee with saphenous vein vs synthetic graft. AB - There is still debate whether sintethic graft (polytetrafluoroethylene or Dacron) is equivalent to vein as bypass graft material for the above-knee femoropopliteal bypass. Therefore, we performed prospective randomized trial to compare vein with polytetrafluoroethylene/dacron for femoropopliteal bypasses with the distal anastomosis above the knee. Between January 2000 and June 2003, 121 femoropopliteal bypasses were performed. The indications for operation were severe claudication in 96 cases, rest pain in 16 cases, and ulceration in 9 cases. After randomization, 60 reversed saphenous venous bypasses and 61 polytetrafluoroethylene/dacron bypasses were performed. No perioperative mortality was seen, and 5% of the patients had minor infections of the wound, not resulting in loss of the bypass, the limb, or life. After 5 years, 37% of the patients had died and 7% were lost to follow-up. Only once saphenous vein was necessary for coronary artery bypass grafting. Primary patency rates after 5 years were 76,6% for venous bypass grafts and 59,1% for polytetrafluoroethylene/dacron grafts (p=0,035). Secondary patency rates were 83,3% for vein and 69,2% for polytetrafluoroethylene/dacron bypasses (p = 0,036). In the venous group, 10 bypasses failed, leading to four new bypasses. In the polytetrafluoroethylene group, 22 bypasses failed, leading to 12 reinterventions. After 5 years of follow-up, we conclude that a bypass with saphenous vein has better patency rates at all intervals and needs fewer reoperations. Saphenous vein should be the graft material of choice for above-knee femoropopliteal bypasses and should not be preserved for reinterventions. Polytetrafluoroethylene/dacron is an acceptable alternative if the saphenous vein is not available. PMID- 19125711 TI - Invasive mole--case report of massive uterine destruction. AB - Patient with malignant Gestational Trophoblastic Neoplasm (GTN) was treated by mean of MTX-FA, MAC, EMA-CO and EMA-EP. Changes in serum human chorionic gonadotropine (beta hCG) levels and changes in ultrasonographic findings were checked weekly. Finally transabdominal hysterectomy with ovaries conservation was done and polychemotherapy administrated after the operation until three consecutive serum chorionic gonadotropine values were negative. This is a case report of Invasive mole in 32 years old patient without possibility to preserve reproductive health. GTN developed two months after spontaneous abortion in 13th week gestation. No changes in uterine structure were found during the first ultrasonographic examination. Three months after abortion and one month after GTN confirmed, massive destruction of lateral uterine wall was detected during transvaginal Doppler ultrasound examination. Resistance index of 0,366 was significantly lower than normal, with hypervascularisation in affected tissue. Serum beta hCG confirmed poor effect of polychemotherapy treatment and decision for operative treatment was made. Hystological findings after the operation confirmed malignant GTN- invasive mole. Specific changes in ultrasonographic picture could have an impact in therapy making decision and could not be refereed without the most relevant parameter such is serum human chorionic gonadotropine. PMID- 19125712 TI - Pattern of head and neck malignant tumours in a Tuzla ENT clinic--a five year experience. AB - Head and neck neoplasia can affect certain fundamental functions, including eating, drinking, speaking and respiration. One overriding factor in deciding on treatment policy is the tendency for head and neck malignancy to be limited to the primary site and regional lymph nodes with surgery and chemotherapy and radiotherapy. The aim of the study was to analyze the occurrence of Head and Neck tumours operated at ENT clinic Tuzla, University Clinical Center Tuzla, Bosnia and Herzegovina. Medical records of patients with histopathologically confirmed head and neck malignancies over a 5 year period (2003-2007) were analyzed. Eight hundred and eight one (881) cases, made up of 519 (58,8%) males and 362 (41,1%) females were found. The most common sites for head and neck malignancies were found to be in the larynx (26,1%), oral cavity (21,7%), the thyroid gland (14,64 %) and the neck (8,51%). A total of 230 patients were diagnosed with laryngeal carcinoma (M:173; F:57), showing the increasing number of female patients. The histopathological tumour types found in this work were mostly squamous cell carcinoma (72,09%), papillary carcinoma (12,2%), while many other minor histopathological variants accounted for 13%. The most patients were presented with stage I and stage III of disease (27% and 28,3%), and 19,9% with stage IV. About 40% of patients did not have data about smoking habit and alcohol drinking. The incidence of head and neck tumours seems to be relatively high, but without significant increase during investigated period. More investigation concerning risk factors, diagnostic procedures, and management strategies should be done in future. PMID- 19125713 TI - In vitro examination of ontogenesis of developing neuronal cells in vagal nuclei in medulla oblongata in newborns. AB - The development of neuron cells in vagal nerve nuclei in medulla oblongata was studied in vitro in live newborns and stillborns from different cases. Morphological changes were studied in respiratory nuclei of dorsal motor centre (DMNV) and nucleus tractus solitarius (NTS) in medulla oblongata. The material from medulla oblongata was fixated in 10 micro buffered formalin solution. Fixated material was cut in series of 10mu thickness, with starting point from obex in +/- 4 mm thickness. Special histochemical and histoenzymatic methods for central nervous system were used: cresyl echt violet coloring, tolyidin blue, Sevier-Munger modification and Grimelius coloring. In immature newborns (abortions and immature) in dorsal motor nucleus of the vagus (DMNV) population stages S1, S2, S3 are dominant. In neuron population in vagal sensory nuclei (NTS) stages S1, S2 are dominant. In more advanced stages of development of newborns (premature), in DMNV stages S3 and S4 are seen and in NTS stages S2 and S3 are dominant. In mature phase of newborns (maturity) in vagal nucleus DMNV stages S5 and S6 are dominant, while in sensory nucleus NTS stages S4 and S5 are dominant. These data suggest that neuron population in dorsal motor nucleus of the vagus (DMNV) are more advanced in neuronal maturity in comparison with sensory neuron population of vagal sensory nucleus NTS. This occurrence shows that phylogenetic development of motor complex is more advanced than the sensory one, which is expected to take new information's from the extra uterine life after birth (extra uterine vagal phenotype). PMID- 19125714 TI - Our experiences with erlotinib in second and third line treatment patients with advanced stage IIIB/ IV non-small cell lung cancer. AB - HeadHER1/EGFR is known to play a pivotal role in tumorigenesis and is overexpressed in up to 80% of NSCLCs. The study of an Expanded Access Clinical Program of Erlotinib in NSCLC is a phase IV open-label, non-randomized, multicenter trial in patients with advanced (inoperable stage IIIb/IV) NSCLC who were eligible for treatment with erlotinib but had no access to trial participation. Patients for the study from Bosnia and Herzegovina (B&H) were selected from two Clinical centres (Sarajevo and Banja Luka). The aim of study was to evaluated efficacy and tolerability of erlotinib monotherapy in this setting. All patients who received at least one dose of erlotinib and data were entered in the database as of the CRF cut-off date of 14th May 2008 were included in analysis of data (n = 19). This population is defined as the Intent to Treat (ITT) population and includes all patients who had at least one dose of erlotinib regardless of whether major protocol violations were incurred. The findings are consistent with the results of the randomized, placebo-controlled BR.21 study. Indicating that erlotinib is an effective option for patients with advanced NSCLC who are unsuitable for, or who have previously failed standard chemotherapy. In B&H group of patients DCR was almost 84%, and PFS was approximately 24,7 weeks (compared with 44% and 9,7 weeks for erlotinib reported in phase III). Almost three quarter of the patients received erlotinib as their second line of therapy. Overall, erlotinib was well tolerated; there were no patients who withdrew due to a treatment-related AE (mainly rash) and there were few dose reductions. 24% of patients experienced an SAE (most commonly gastrointestinal (GI) disorders). PMID- 19125715 TI - Comparison of some physical parameters of whole and scored lisinopril and lisinopril/hydrochlorthiazide tablets. AB - Tablets are one of the most popular and preferred solid dosage forms because they can be accurately dosed, easily manufactured and packaged on a large scale, have good physical and chemical stability, and can contribute to good patient compliance given their ease of administration. The ability to match doses to patients depends on the availability of multiple dose sizes and adequate dose response information. These are not always provided, so splitting of the tablets is sometimes necessary. Tablet splitting is an accepted practice in dispensing medication. It has been used when a dosage form of the required strength is not available commercially. The aim of our study was to compare some physical parameters of whole and scored lisinopril and lisinopril/hydrochlorthiazide tablets and to accept or exclude their influence on the obtaining of required dosage. According to the results obtained, we may conclude that tablets from batch "I", "II", "III" and "IV" satisfied pharmacopeial requirements concerning crushing strength, friability, disintegration time and mass uniformity. The hardness testing showed acceptable reproducibility and indicate that the data variation was primarily from the irreversible changes in the structure of tablet samples. The act of compacting powders stores energy within the tablets, by shifting or compressing the intermolecular bonds within the particles. The tablets have a natural tendency to relax once pressure is removed, and this tendency works against the interparticle bonding formed during compression. Hardness testing procedure causes irreversible changes in this structure. PMID- 19125717 TI - Dendritic cells: biology of the skin. AB - Allergic contact dermatitis results from a T-cell-mediated, delayed-type hypersensitivity immune response induced by allergens. Skin dendritic cells (DCs) play a central role in the initiation of allergic skin responses. Following encounter with an allergen, DCs become activated and undergo maturation and differentiate into immunostimulatory DCs and are able to present antigens effectively to T cells. The frequency of allergic skin disorders has increased in the past decades. Therefore, the identification of potential sensitizing chemicals is important for skin safety. Traditionally, predictive testing for allergenicity has been conducted in animal models. For regulatory reasons, animal use for sensitization testing of compounds for cosmetic purposes is shortly to be prohibited in Europe. Therefore, new non-animal-based test methods need to be developed. Several DC-based assays have been described to discriminate allergens from irritants. Unfortunately, current in vitro methods are not sufficiently resilient to identify allergens and therefore need refinement. Here, we review the immunobiology of skin DCs (Langerhans' cells and dermal dendritic cells) and their role in allergic and irritant contact dermatitis and then explore the possible use of DC-based models for discriminating between allergens and irritants. PMID- 19125718 TI - Read-across to rank skin sensitization potential: subcategories for the Michael acceptor domain. AB - BACKGROUND: Eliminating animal testing for skin sensitization is a significant challenge in consumer safety risk assessment. To be able to perform resilient risk assessments in the future, one will need alternative approaches to fill the data gaps. OBJECTIVES: To this end, we propose a subcategory-based read-across approach to estimate and rank skin sensitization potential of chemicals. The example described here is for the mechanism of Michael-type nucleophilic addition with subcategories being limited to carbonyl-containing compounds. PATIENTS/METHODS: In this approach, in silico tools based on structural alerts were used to determine both the mechanism of protein binding and the relative subcategories within that mechanism. RESULTS: Fifty compounds previously evaluated in the in vivo mouse local lymph node assay (LLNA) were placed in 10 subcategories defined by their polarized alpha,beta-unsaturated substructure. To offset the limitations and skewness of the published in vivo data, in chemico glutathione (GSH) depletion data also were included. CONCLUSIONS: It was shown that the read-across approach can be successfully used to rank qualitatively skin sensitization potential of an untested carbonyl-containing Michael acceptor chemical by using subcategories. Moreover, the use of the more resilient in chemico GSH depletion data added further support to the read-across result. PMID- 19125719 TI - Air oxidation increases skin irritation from fragrance terpenes. AB - BACKGROUND: Linalool and limonene are common fragrance terpenes that autoxidize on air exposure. The pure compounds are not allergenic but their oxidation products can cause contact allergy. Little has been investigated regarding the irritancy of oxidized terpenes. AIM: The aim of this study was to investigate the irritating effect of pure and oxidized R-limonene and linalool in concentration series and to study the MNIC (Maximum Non Irritant Concentration) of autoxidized linalool and limonene. PATIENTS/METHODS: Patch testing was performed in dermatitis patients and controls with sequentially diluted concentrations of oxidized and non-oxidized linalool, and oxidized and non-oxidized R-limonene. Readings were made with visual assessment and using laser Doppler imaging. RESULTS: The non-oxidized terpenes were non-irritating in all tested concentrations. Both linalool and especially R-limonene were more irritating after oxidation compared with the pure compounds. No difference in response was seen between dermatitis patients and controls. CONCLUSION: Autoxidation of the fragrance terpenes linalool and R-limonene increases irritation. Oxidized linalool is less irritating than oxidized R-limonene. In this study, we found no advantages in using laser Doppler technique compared with visual assessment. PMID- 19125721 TI - Occupational airborne allergic contact dermatitis in a concrete repair worker. PMID- 19125720 TI - There is an association between contact allergy to aluminium and persistent subcutaneous nodules in children undergoing hyposensitization therapy. AB - BACKGROUND: The development of persistent itchy nodules at the injection site following hyposensitization therapy with aluminium-precipitated antigen extract has been described in several reports. Occasionally, contact allergy to aluminium has been reported in individuals with such nodules. OBJECTIVES: To investigate if hyposensitization therapy can induce contact allergy to aluminium and examine if there is any association between persistent subcutaneous nodules and aluminium allergy. PATIENTS/METHODS: Sixty-one children with allergic asthma and/or allergic rhinitis participated in the study of whom 37 had had hyposensitization therapy. The study consisted of a non-clinical part based on a questionnaire and a clinical part with a physical examination, self-assessment of itching, and patch testing. To secure an unbiased evaluation of possible reactions, the investigators were blinded. RESULTS: Contact allergy to aluminium was found in eight participants, all in the exposed group (8/37 versus 0/24, P = 0.02). Examination showed nodules on the upper arms in 13 participants, all in the group exposed to hyposensitization therapy. Nodules were over-represented in patients with contact allergy to aluminium. CONCLUSIONS: There was a statistically significant association between contact allergy to aluminium and persistent subcutaneous nodules in children who had had hyposensitization therapy. PMID- 19125722 TI - Occupational airborne allergic contact dermatitis from potassium metabisulfite. PMID- 19125723 TI - Lymphomatoid contact dermatitis. PMID- 19125724 TI - Scratch-patch test for the diagnosis of allergic contact dermatitis to aciclovir. PMID- 19125725 TI - Increased serum levels of IL-22 in patients with nickel contact dermatitis. PMID- 19125726 TI - Active sensitization to chloracetamide. PMID- 19125727 TI - Allergic contact dermatitis due to mono(2-ethylhexyl) maleate in di-(n-octyl)tin bis(2-ethylhexyl maleate) in polyvinyl chloride gloves. PMID- 19125728 TI - Concordance of patch test results with four new TRUE test allergens compared with the same allergens from Chemotechnique. PMID- 19125729 TI - Mercury-induced systemic allergic dermatitis caused by 'white precipitate' in a skin lightening cream. PMID- 19125730 TI - Contact allergy to polyolefin ester in a lubricant grease. PMID- 19125731 TI - Whole-body vibration dosage alters leg blood flow. AB - The effect of whole-body vibration dosage on leg blood flow was investigated. Nine healthy young adult males completed a set of 14 random vibration and non vibration exercise bouts whilst squatting on a Galileo 900 plate. Six vibration frequencies ranging from 5 to 30 Hz (5 Hz increments) were used in combination with a 2.5 mm and 4.5 mm amplitude to produce twelve 1-min vibration bouts. Subjects also completed two 1-min bouts where no vibration was applied. Systolic and diastolic diameters of the common femoral artery and blood cell velocity were measured by an echo Doppler ultrasound in a standing or rest condition prior to the bouts and during and after each bout. Repeated measures MANOVAs were used in the statistical analysis. Compared with the standing condition, the exercise bouts produced a four-fold increase in mean blood cell velocity (P<0.001) and a two-fold increase in peak blood cell velocity (P<0.001). Compared to the non vibration bouts, frequencies of 10-30 Hz increased mean blood cell velocity by approximately 33% (P<0.01) whereas 20-30 Hz increased peak blood cell velocity by approximately 27% (P<0.01). Amplitude was additive to frequency but only achieved significance at 30 Hz (P<0.05). Compared with the standing condition, squatting alone produced significant increases in mean and peak blood cell velocity (P<0.001). The results show leg blood flow increased during the squat or non vibration bouts and systematically increased with frequency in the vibration bouts. PMID- 19125732 TI - Alterations in VOmax and the VO plateau with manipulation of sampling interval. AB - BACKGROUND: The most accepted criterion for confirming attainment of VO(2)max is a plateau in oxygen consumption (VO(2)) at VO(2)max, but its incidence varies. AIMS: To compare VO(2)max and VO(2) plateau incidence across various sampling intervals, and to examine predictors of the change in VO(2) (DeltaVO(2)) at VO(2)max. METHODS: Sedentary, recreationally-active, and endurance-trained subjects (n = 108, age = 24.2 +/- 6.2 year) completed incremental exercise on the treadmill or cycle ergometer. Gas exchange data were obtained breath-by-breath and time-averaged every 15, 30, and 60 s. VO(2)max attainment was verified with the Taylor et al. (1955) criterion (DeltaVO(2) at VO(2)max < or =2.1 ml kg(-1) min(-1)). Multiple regression was used to examine predictors of DeltaVO(2) at VO(2)max. RESULTS: VO(2) plateau incidence was higher using breath-by-breath (81%) and 15 (91%) and 30 s time averaging (89%) versus 60 s averaging (59%). Compared to 60 s averaging, VO(2)max was significantly higher (P<0.05) when data were obtained breath-by-breath and with 15 and 30 s time-averaging compared to 60 s sampling. VO(2)max was not related to VO(2) plateau incidence. Respiratory rate was a significant predictor of DeltaVO(2) at VO(2)max in endurance-trained subjects. CONCLUSION: More frequent data acquisition revealed higher VO(2)max and incidence of the VO(2) plateau compared to 60 s time averaging. Secondary criteria to verify VO(2)max attainment should not be used, as they do not discern between subjects who do and do not reveal a plateau in VO(2) at VO(2)max. PMID- 19125733 TI - Bif-1 and Bax expression in cutaneous Merkel cell carcinoma. AB - BACKGROUND: Bax-interacting factor-1 (Bif-1) binds to Bax, which in turn activates this proapoptotic protein. In the absence of Bif-1, the ability to induce apoptosis through the intrinsic pathway is greatly reduced. Merkel cell carcinoma (MCC) classically shows an aggressive behavior and lack of response to chemotherapy, which remains unexplained. Previous studies have documented the presence of Bax in MCC, but Bif-1 expression has not been evaluated. Herein, the expression of Bif-1 and Bax in cutaneous MCC is examined. MATERIALS AND METHODS: The immunohistochemical expression of Bif-1 and Bax protein was examined in nine cases of MCC. Both positive and negative controls were conducted. All the cases were reviewed by a single dermatopathologist. RESULTS: Bif-1 was detected in nine cases (100%), and Bax was expressed in six cases (66%). The percent positive cells for Bif-1 in MCC ranged from 85% to 98% positive (mean 93.9%). At the same time, decreased Bax expression was shown with 0-8% positive cells (mean 3.45%). CONCLUSION: The increased expression of Bif-1 in MCC is associated with low levels of Bax staining. These findings suggest that the upregulation of Bif-1 could in part be responsible for tumorigenesis in cutaneous MCC. As shown, Bax and Bif-1 expression are not exclusively antithetical; therefore, future studies evaluating the expression of both proteins should be conducted. PMID- 19125734 TI - Primary cutaneous spindle cell B-cell lymphoma with multiple figurate erythema like manifestation. AB - We report a 68-year-old Korean man presenting with asymptomatic erythematous polycyclic annular firm plaques on his back that spread to the right shoulder. Histopathologic examination showed dense, diffuse infiltrates involving the entire dermis, consisting of atypical lymphocytes with many centrocytes and a few centroblasts. Spindle-shaped cells with elongated, twisted nuclei containing dispersed chromatin were also seen. Immunohistochemical analysis showed that all of the cells were strongly positive for CD20, CD21, CD79a and CD45, while they were negative for CD3, CD5, CD10, CD23, CD35, CD43, CD45RO and CD68. The spindle cells were also negative for smooth-muscle actin, desmin, S-100 and CD34. They consistently expressed nuclear bcl-6, but did not express bcl-2, multiple myeloma 1 and p16. We diagnosed him with primary cutaneous spindle cell B-cell lymphoma (PCSBCL) and treated him with six cycles of cyclophosphamide, adriamycin, vincristine, prednisone and rituximab (R-CHOP) chemotherapy; his skin lesions disappeared completely. Immunohistochemical profiles suggest that PCSBCL is a variant of primary cutaneous follicle center lymphoma. PMID- 19125735 TI - Rhabdomyomatous mesenchymal hamartoma presenting on a digit. AB - Rhabdomyomatous mesenchymal hamartoma is a rare dermal lesion comprised of skeletal muscle, adipocytes and collagen. The vast majority has been described in very young patients on the head and neck. To our knowledge, there are only two reports of rhabdomyomatous mesenchymal hamartoma outside the head and neck region. Both of those lesions were located in the perianal area. We describe a rhabdomyomatous mesenchymal hamartoma in a 36-year-old man located on the great toe. Although rhabdomyomatous mesenchymal hamartoma is uncommon, it is important to be aware of this entity, its possible association with congenital syndromes and its potential for localization outside of the head and neck region. PMID- 19125736 TI - Ossifying fibromyxoid tumor of soft parts mimicking a schwannoma with uncommon histology: a potential diagnostic pitfall. PMID- 19125737 TI - Squamous-cell carcinoma in situ is a fiction! PMID- 19125739 TI - Development of an in vitro Bowen's disease model using a three-dimensional culture system. PMID- 19125740 TI - CD34-positive myxoid dermatofibrohistiocytoma of the skin: an indolent post traumatic tumor that can be mistaken for dermatofibrosarcoma protuberans. PMID- 19125741 TI - Radiotherapy-associated atypical vascular lesions of the breast. PMID- 19125744 TI - New Zealand hosts 12th Scientific Meeting in Optometry. PMID- 19125742 TI - CD34-reactive tumors of the skin. An updated review of an ever-growing list of lesions. AB - Over the past few years, a growing number of cutaneous tumors expressing CD34 is being reported. The list contains benign and malignant neoplasms as well as reactive and hamartomatous lesions of diverse lineages of differentiation, including fibroblastic, myofibroblastic, fibrohistiocytic, vascular, neural, adipocytic, smooth muscle, hematopoietic, melanocytic and epithelial. The more frequent diagnostic difficulties are found in spindle cell proliferations, mainly in those of the fibrocytic lineage. In part, this is because of the fact that in this area are, aside to well-defined entities, histologically and clinically diverse, recently reported cutaneous CD34-reactive lesions, whose definitions, limits and relationships are not completely established. The CD34 expression plays a key role in the differential diagnosis of some tumors, such as dermatofibrosarcoma protuberans, epithelioid sarcoma or pleomorphic hyalinizing angiectatic tumor of soft parts, with important therapeutic consequences. In others, as in desmoplastic trichilemmoma, it can help to resolve diagnostic problems in concrete cases. Finally, in many of the CD34-positive lesions, the diagnosis with the hematoxylin and eosin stain is straightforward. However, in all of them, the knowledge of the immunohistochemical profile contributes to our understanding of the cutaneous pathology. PMID- 19125745 TI - Rapid resolution of severe retinal neovascularisation in proliferative diabetic retinopathy following adjunctive intravitreal bevacizumab (Avastin). AB - A 23-year-old aboriginal woman with severe bilateral proliferative diabetic retinopathy and left rubeotic glaucoma received bilateral pan-retinal photocoagulation with adjunctive intravitreal bevacizumab. Six days post injection, there was almost complete resolution of new vessels. PMID- 19125746 TI - Raising awareness of blindness as another smoking-related condition: a public health role for optometrists? PMID- 19125747 TI - Rapid confrontation screening for peripheral visual field defects and extinction. AB - Screening for unsuspected visual field defects should form a part of all routine eye examinations. Here, we review a procedure for finger-counting confrontation screening that tests the periphery of all visual field quadrants of each eye, yet requires a total of only four responses from the patient. In addition, the test simultaneously screens for the extinction phenomenon that can accompany unilateral brain damage. Due to its efficiency, we recommend that this procedure form the standard way that screening finger-counting confrontation be performed, with abnormal findings prompting a more detailed assessment of visual fields and further neurological examination as necessary. Our paper is not intended to suggest that finger-counting confrontation is superior to other forms of visual field screening and indeed the literature suggests its sensitivity is limited. PMID- 19125753 TI - NICE guidance on continuous subcutaneous insulin infusion 2008: review of the technology appraisal guidance. PMID- 19125754 TI - The importance of health belief models in determining self-care behaviour in diabetes. AB - Patients' self-care behaviours have a major role in diabetes management. Diabetes education provides the required knowledge, but despite this, self-care is often suboptimal. The degree to which patients follow advice as regards the various self-care behaviours is determined by their health beliefs (Illness Representations or Personal Models) of diabetes. Psychometric studies have tried to categorize and measure the beliefs about illness that influence patients to adhere to treatment recommendations in diabetes. Various models have been proposed to explain the relationship between beliefs and behaviour. Leventhal's Self-Regulatory Model, which takes account of the emotional as well as the objective rational response to illness, currently seems to offer the best system for identifying the determinants of patient self-care behaviour. A review of interventions indicates those based on psychological theory offer professionals the best chance of maximizing their patients' contribution to diabetes self management and achieving improved outcomes, both glycaemic and psychosocial. Studies designed specifically to modify illness representations are now being undertaken. This brief review aims to summarize developments in this area of psychological theory over the last 20 years and the implications for promoting better self-care behaviour in diabetes. PMID- 19125755 TI - Pregnancy outcome in patients with raised blood glucose due to a heterozygous glucokinase gene mutation. AB - AIM: To assess determinants of fetal growth in the offspring of pregnant women with hyperglycaemia due to a heterozygous glucokinase (GCK) gene mutation. METHODS: Details of gestational age at delivery, fetal birth weight and maternal antenatal treatment were collected from patients and retrospective case note review of 82 offspring born to 42 women with GCK gene mutations and 31 offspring born to 13 unaffected normoglycaemic women with an affected partner. Fetal genotype was determined using direct sequencing from either a mouth swab or a blood sample. RESULTS: In mothers with GCK mutations, non-mutation-carrying offspring were heavier than mutation-carrying offspring (corrected birth weight 3.9 +/- 0.6 vs. 3.2 +/- 0.8 kg; P < 0.001) and more likely to be macrosomic (> 4.0 kg; 39% vs. 7%, P = 0.001). There was no difference in corrected birth weight between offspring of insulin- and diet-treated women (3.7 +/- 0.7 vs. 3.8 +/- 0.6 kg; P = 0.1), although insulin-treated mothers delivered earlier (37.5 +/- 1.7 vs. 38.9 +/- 2.3 weeks; P < 0.001) due to increased obstetric intervention. CONCLUSIONS: Offspring of women with GCK mutations are at increased risk of macrosomia and its obstetric consequences. Fetal birth weight is predominantly altered by fetal genotype and not treatment of maternal hyperglycaemia with insulin. This probably reflects the large effect of a fetal GCK mutation on fetal insulin secretion and the difficulty in reducing the regulated maternal glycaemia caused by a glucose sensing defect in people with GCK mutations. PMID- 19125756 TI - A double-blind, randomized, placebo-controlled trial of the short-term effect of vitamin D3 supplementation on insulin sensitivity in apparently healthy, middle aged, centrally obese men. AB - AIM: To determine the short-term effect of vitamin D(3) supplementation on insulin sensitivity in apparently healthy, middle-aged, centrally obese men. SUBJECTS AND METHODS: A double-blind randomized controlled trial was conducted at a tertiary care facility in which 100 male volunteers aged > or = 35 years received three doses of vitamin D(3) (120,000 IU each; supplemented group) fortnightly or placebo (control group). Hepatic fasting insulin sensitivity [homeostasis model assessment (HOMA), quantitative insulin-sensitivity check index, HOMA-2], postprandial insulin sensitivity [oral glucose insulin sensitivity (OGIS)], insulin secretion (HOMA%B, HOMA2-%B), lipid profile and blood pressure were measured at baseline and at 6 weeks' follow-up. RESULTS: Seventy-one of the recruited subjects completed the study (35 in supplemented group, 36 in control group). There was an increase in OGIS with supplementation by per protocol analysis (P = 0.038; intention-to-treat analysis P = 0.055). The age- and baseline 25-hydroxyvitamin D level-adjusted difference in change in OGIS was highly significant (mean difference 41.1 +/- 15.5; P = 0.01). No changes in secondary outcome measures (insulin secretion, basal indices of insulin sensitivity, blood pressure or lipid profile) were found with supplementation. CONCLUSION: The trial indicates that vitamin D(3) supplementation improves postprandial insulin sensitivity (OGIS) in apparently healthy men likely to have insulin resistance (centrally obese but non-diabetic). PMID- 19125757 TI - Symptoms of depression and diabetes-specific emotional distress are associated with a negative appraisal of insulin therapy in insulin-naive patients with Type 2 diabetes mellitus. A study from the European Depression in Diabetes [EDID] Research Consortium. AB - AIMS: A meta-analysis concluded that depression is associated with poor glycaemic control in Type 2 diabetes (DM2). In DM2 patients with deteriorating glycaemic control, the initiation of insulin therapy is often postponed. The aim of the present study was to determine whether symptoms of depression and diabetes specific emotional distress are associated with a more negative appraisal of insulin therapy. METHODS: We collected cross-sectional data in two outpatient university clinics in Istanbul, Turkey. The study sample consisted of 154 insulin naive patients with DM2. A self-report questionnaire was used to obtain demographic and clinical data. Main instruments were the Centre for Epidemiologic Studies Depression Scale, (CES-D), the Problem Areas In Diabetes scale (PAID) and the Insulin Treatment Appraisal Scale (ITAS). RESULTS: Analysis of variance revealed that patients with a higher depression score rated insulin therapy significantly more negative then patients with lower depression scores. Moreover, 47% of patients with a high depression score had a negative appraisal of insulin therapy on 7 or more of the 20 ITAS-items, compared to 25 to 29% of those with low-moderate depression scores. Multiple regression analyses showed that a negative appraisal of insulin therapy was significantly associated with higher depression and diabetes-distress scores and low education, but not with sex, age or duration of diabetes. CONCLUSIONS: Our results suggest that in insulin-naive Type 2 diabetes patients, higher levels of depression and diabetes-distress tend to be associated with more negative beliefs about insulin. Whether these negative attitudes translate into postponing initiation of insulin therapy needs to be tested in longitudinal research. PMID- 19125758 TI - Blood pressure control and awareness among patients with diabetes and hypertension attending a tertiary ophthalmic clinic. AB - AIMS: To assess the proportion of patients who had diabetes and hypertension with optimal blood pressure (BP) control and to describe patients' awareness of the importance of BP control. METHODS: A cross-sectional survey of out-patients with diabetes who attended the Royal Victorian Eye and Ear Hospital (Melbourne, Australia) was conducted between October 2006 and February 2007. Of 245 patients invited, 220 (90%) participated. Optimal BP control was defined as BP < 130/80 mmHg recorded at the latest visit; proportions of patients with awareness of BP control were defined non-exclusively by: (i) ability to recall a previous BP record and rate the appropriateness of that BP level; (ii) ability to recall the recommended optimal BP control level by current diabetes management guidelines; (iii) knowledge that optimal BP control is important to diabetes management; and (iv) knowledge that optimal BP control is important to eye health. RESULTS: Of the 220 patients, 176 had both diabetes and hypertension. Of these, 49 of 176 (28%) had BP controlled optimally, and 30 of 176 (17%) recalled and rated a previous recorded BP level appropriately. Fewer than one in four (22%) acknowledged the recommended optimum BP level of < 130/80 mmHg, fewer than one in two (48%) rated BP as important to diabetes management, and one in three (35%) rated BP control as important to eye health. CONCLUSIONS: In this sample of patients with diabetes and hypertension, fewer than one-third achieved the BP level recommended by clinical guidelines, and fewer than half were aware of the importance of BP control. PMID- 19125759 TI - Examining the effectiveness of a peer-led education programme for Type 2 diabetes and cardiovascular disease in a Bangladeshi population. AB - AIMS: The aim of this study was to assess an adapted version of the X-PERT Programme for Type 2 diabetes within a Bangladeshi population. METHODS: Bangladeshi adults, living in the UK, with Type 2 diabetes participated in a diabetes educational session based on an adapted version of the X-PERT Programme. Participants attended a session, led by a trained peer educator and carried out in Sylheti. All participants who registered on the course were telephoned or visited in person the day before the course as a reminder. RESULTS: Registration to attend the course was excellent. However, actual attendance rates were 58% (42/72) out of those registered. Once participants attended, overall they enjoyed attending the sessions and felt they benefited as they understood more about how to better self-manage their condition. Those aspects of the sessions that were most enjoyed were group discussions and interactive posters to explain diabetes. Participants requested a home-based exercise guide, which was developed by the research team. The study was underpowered to show behaviour change, but did demonstrate a trend to improvement in self-care activities. All participants reported recommending the course to others. CONCLUSIONS: There was an excellent response in terms of registration for the course. However, the time to attend the course appeared to be a barrier to participants. Nevertheless, for those attending the course, responses were very positive. The amended X-PERT Programme could be used as a component of a package to improve outcome and self-management for people with diabetes in the Bangladeshi community. PMID- 19125760 TI - Prevalence and predictors of osteopenia and osteoporosis in adults with Type 1 diabetes. AB - AIMS: To determine the prevalence and biochemical/hormonal determinants of osteopenia and osteoporosis in adults with Type 1 diabetes. METHODS: One hundred and two patients (52 female, 50 male) with Type 1 diabetes aged 20-71 years underwent cross-sectional assessment of biochemical/hormonal markers of bone metabolism, and bone mineral density (BMD) measurement at forearm, hip and spine using dual energy x-ray absorptiometry. BMD data were available for 102 age- and gender-matched population-based control subjects. RESULTS: After adjusting for age and body mass index (BMI), osteopenia and osteoporosis were more common at the spine in males with Type 1 diabetes than in control subjects (P = 0.030). In Type 1 males, after adjustment for age and BMI, BMD, T- and Z-scores at the hip, femoral neck and spine were lower compared with age-matched control subjects (P < or = 0.048). Female Type 1 patients and control subjects had similar BMDs and T- and Z-scores at all sites. On multiple linear regression analysis, which adjusted for the natural logarithm of the sex hormone binding globulin concentration, smoking status and alcohol consumption, and (for women) menopausal status, each of BMI, serum ionized calcium and serum alkaline phosphatase (negatively) were independently associated with BMD at the hip and femoral neck in Type 1 diabetic subjects. CONCLUSIONS: Adult males with Type 1 diabetes have reduced bone density at the hip, femoral neck and spine when compared with age-matched control subjects. Impaired bone formation may occur in Type 1 diabetes. PMID- 19125761 TI - Risk factor control in patients with Type 2 diabetes and coronary heart disease: findings from the Swedish National Diabetes Register (NDR). AB - AIMS: Patients with Type 2 diabetes and coronary heart disease (CHD) are infrequently treated to risk factor targets in current guidelines. We aimed to examine risk factor management and control levels in a large sample of patients with Type 2 diabetes with CHD. METHODS: This was an observational study of 1612 patients with first incidence of CHD before 2002, and of 4570 patients with first incidence of CHD before 2005, from the Swedish National Diabetes Register (NDR). RESULTS: In patients with CHD 1-2 years before follow-up, the achievement of cardiovascular risk factor targets (follow-up 2002/follow-up 2005) was: HbA(1c) < 7%, 47%/54% (P < 0.01); blood pressure < or = 130/80 mmHg, 31%/40% (P < 0.001); total cholesterol < 4.5 mmol/l, 47%/60% (P < 0.001); and low-density lipoprotein cholesterol < 2.5 mmol/l, 49%/65% (P < 0.001). Use of medication: antihypertensives, 90%/94% (P < 0.01); lipid-lowering drugs, 75%/86% (P < 0.001); and aspirin, 85%/89% (P < 0.05). A high prevalence of adverse lifestyle characteristics prevailed (2002/2005): overweight [body mass index (BMI) > or = 25 kg/m(2)], 86%/85%; obesity (BMI > or = 30 kg/m(2)), 41%/42%; smokers in age group < 65 years, 16-23%/18-19%; as well as waist circumference > or = 102 cm (men) or > or = 88 cm (women), 68% in 2005. CONCLUSIONS: Patients with a combination of Type 2 diabetes and CHD showed an increased use of lipid-lowering drugs over time, corresponding to improving blood lipid levels. A discrepancy existed between the prevalent use of antihypertensive drugs and the low proportion reaching blood pressure targets. Regretfully, a high prevalence of adverse lifestyle characteristics prevailed. Evidence-based therapy with professional lifestyle intervention and drugs seems urgent for improved quality of secondary prevention in these patients. PMID- 19125762 TI - Healthcare charges and utilization associated with diabetic neuropathy: impact of Type 1 diabetes and presence of other diabetes-related complications and comorbidities. AB - AIMS: The aim was to examine the impact of Type 1 diabetes and having any other diabetes-related complication or comorbidity on healthcare charges and utilization in patients with diabetic neuropathy (DN). METHODS: We selected individuals aged < 65 years who continuously enrolled in a large US commercial plan from July 2004 to June 2006 and who received at least one diagnosis of DN at any time from July 2004 to June 2005. We compared the prevalence of other diabetes-related complications or comorbidities between patients with Type 1 and with Type 2 diabetes. In patients with DN with or without any other diabetes related complication or comorbidity, we used multivariate regression to assess the marginal contribution of Type 1 diabetes on healthcare charges and utilization from July 2005 until June 2006. RESULTS: The majority of DN patients had at least one other diabetes-related complication or comorbidity. Most of the DN patients had Type 2 diabetes. DN patients with Type 1 diabetes had more comorbid medical conditions than those with Type 2 diabetes. Compared with Type 2, Type 1 patients had a higher prevalence of each individual non-DN diabetes related complication or comorbidity, except heart disease. Controlling for comorbidities, Type 1 and Type 2 patients with DN but no other diabetes-related complication or comorbidity had similar healthcare utilization. However, Type 1 patients had significantly higher charges than those with any other diabetes related complication or comorbidity. CONCLUSIONS: Many patients with DN have Type 1 diabetes and other common diabetes-related complications or comorbidities, which can have a significant impact on healthcare charges and utilization. PMID- 19125763 TI - Antiplatelet therapy, Helicobacter pylori infection and complicated peptic ulcer disease in diabetes: the Fremantle Diabetes Study. AB - AIMS: To assess whether, based on its relationship with complications of peptic ulcer disease (PUD), directed Helicobacter pylori serological screening is justified in diabetic patients prior to commencement of antiplatelet therapy. METHODS: We analysed data from the longitudinal, community-based Fremantle Diabetes Study (FDS). The present substudy included (i) 1301 patients (91.2% of the total FDS sample; mean age 62.0 +/- 13.3 years, 49.5% male) with available sera from baseline assessment between 1993 and 1996, and (ii) a subset of 40 patients admitted to hospital for complicated PUD (bleeding and/or perforation) between baseline and end of June 2006. All hospital admissions for complicated PUD in the population of Western Australia were identified over the same period. Helicobacter pylori IgG antibodies were measured in all patients at baseline and in the subset at the FDS visit prior to hospital admission. RESULTS: Helicobacter pylori seropositivity was present in 60.6% of FDS patients at baseline and was independently associated with increasing age and non-Anglo-Celt/non-Asian ethnicity. There were 2.9 (95% confidence interval 2.1, 3.9) first admissions for complicated PUD per 1000 patient-years, an incidence more than seven times that in the local general population. Independent baseline predictors of hospital admission were increasing age, serum urea, non-aspirin anticoagulant therapy, sulphonylurea therapy, peripheral arterial disease and diabetic retinopathy, but not aspirin use, H. pylori seropositivity or their interaction. CONCLUSIONS: There are diabetes-specific risk factors for complicated PUD, including sulphonylurea use and vascular complications. Knowledge of H. pylori serological status does not predict complicated PUD in diabetes regardless of use of antiplatelet therapy. PMID- 19125764 TI - Mechanism of disopyramide-induced hypoglycaemia in a patient with Type 2 diabetes. AB - BACKGROUND: Disopyramide, an antiarrhythmia drug, has been reported to cause hypoglycaemia. Pre-existing factors that increase the concentration of the drug in the blood increase the risk of hypoglycaemia. Furthermore, other factors can also increase the risk of hypoglycaemia even when disopyramide levels are in the therapeutic range. It has been proposed that disopyramide-induced hypoglycaemia is caused by inhibition of the pancreatic B-cell K(ATP) channels. CASE REPORT: We report a case of severe disopyramide-induced hypoglycaemia in a 62-year-old woman with Type 2 diabetes taking low-dose glimepiride treatment. She had not experienced hypoglycaemia prior to the start of disopyramide therapy. No further hypoglycaemic episodes occurred following withdrawal of disopyramide therapy. FUNCTIONAL STUDY: Current recordings of K(ATP) channels expressed in Xenopus oocytes showed that at their estimated therapeutic concentrations, disopyramide and glimepiride inhibited K(ATP) channels by about 50-60%. However, when both drugs were applied together, K(ATP) channels were almost completely closed (approximately 95%). Such dramatic inhibition of K(ATP) channels is sufficient to cause B-cell membrane depolarization and stimulate insulin secretion. CONCLUSIONS: Disopyramide therapy is not recommended for patients treated with K(ATP) channel inhibitors. PMID- 19125765 TI - Severe insulin resistance due to anti-insulin antibodies: response to plasma exchange and immunosuppressive therapy. AB - Anti-insulin antibodies have been described in two contexts: in insulin-naive individuals (so-called 'insulin autoimmune syndrome') and in patients with insulin-treated diabetes, in whom antibodies are rarely of clinical significance. We report the case of an 68-year-old woman who exhibited a local allergic reaction to subcutaneous insulin followed by severe insulin resistance, evidenced by poor glycaemic control despite treatment with > 3.5 U/kg of insulin per day. She was found to have circulating polyclonal anti-insulin antibodies of the IgG subtype and responded clinically to a course of plasma exchange and immunosuppression with mycophenolate mofetil and, subsequently, intravenous immunoglobulin. Falling titres of antibodies on this regimen correlated with improved glycaemic control. This case suggests that clinicians should be alert to the possibility of insulin resistance due to anti-insulin antibodies and that immunosuppression in this situation may be a valuable therapeutic option. PMID- 19125766 TI - Relationship between post-occlusive forearm skin reactive hyperaemia and vascular disease in patients with Type 2 diabetes--a novel index for detecting micro- and macrovascular dysfunction using laser Doppler flowmetry. AB - AIMS: To investigate the relationship between the novel post-occlusive forearm skin reactive hyperaemia (PORH) index and macro- and microangiopathy by comparing it with traditional indices and a morphological analysis of skin blood flow after ischaemia. METHODS: We studied 20 control subjects and 104 patients with Type 2 diabetes. Cutaneous blood flow (flux) in response to 5-min arterial occlusion in the forearm was measured using laser Doppler flowmetry. We defined the PORH index as the ratio of the area under the curve (AUC) 1 min after the release of the cuff relative to the AUC of a 1-min period before cuff inflation. The flux in response to arterial occlusion was divided morphologically into three patterns. Brachial-ankle pulse wave velocity (baPWV) was measured. RESULTS: The PORH index was markedly lower in patients with diabetes than in control subjects. The PORH index correlated negatively with urinary albumin, peroneal motor nerve conduction velocity, and baPWV in the patients with diabetes. The PORH index, but not the morphological analysis, was an independent determinant of cardiovascular disease in patients with diabetes. CONCLUSIONS: The PORH index may be the most sensitive indicator of micro- and macrovascular disease of the various measures of PORH in patients with Type 2 diabetes. PMID- 19125767 TI - Haemodialysis is associated with changes in cutaneous microcirculation in diabetes mellitus. AB - AIMS: To examine the cutaneous microcirculation on the dorsum of the foot before, during and after haemodialysis in diabetic and non-diabetic patients. METHODS: Fourteen age-matched patients (seven diabetic, seven non-diabetic) without active foot ulceration were studied. Cutaneous microcirculation was assessed using a micro-lightguide spectrophotometer to measure venous oxygen saturation and relative blood flow determined at two tissue depths: 2 and 6 mm. Cumulative relative changes of each parameter during haemodialysis were calculated as area under the curve. Differences between and within the groups were calculated by Mann-Whitney U-test and anova following post hoc testing, respectively. RESULTS: At baseline, relative blood flow at 6 mm tissue depth was significantly greater in diabetic patients (P = 0.048). Thirty minutes after the end of dialysis, relative blood flow at 2 and 6 mm tissue depth was significantly higher in non diabetic patients (P = 0.048 and P = 0.001). Mean cumulative relative changes in venous oxygen saturation and relative blood flow at 2 mm as well as 6 mm tissue depth were positive for non-diabetic subjects and negative for diabetic patients. CONCLUSIONS: Haemodialysis is associated with changes in cutaneous microcirculation, which differ between people with and without diabetes. In those without diabetes, we found an increase in blood flow during haemodialysis, whereas blood flow was reduced in diabetic patients. This may be the result of abnormal vasomotor regulation due to distal neuropathy. PMID- 19125768 TI - An assessment of the accuracy and usability of a novel optical wound measurement system. AB - AIMS: Measurement of wound size can predict healing and provide information to guide treatment. This study assesses a novel optical wound imaging system that creates a three-dimensional image of the ulcer. METHODS: Using a new camera-based digital system and traditional elliptical wound measurements, 36 foot ulcers from 31 patients (aged 44-94 years, median 70 years) were examined during a 12-week period at two centres. Median diabetes duration was 18 years (range 6-56 years). Seventeen percent had Type 1 diabetes, 93% had peripheral neuropathy and 57% had peripheral artery disease. Twenty-five were reviewed consecutively, resulting in 76 ulcer examinations. Median ulcer size was 94 mm(2), with size ranging from 3.1 to 2195 mm(2). RESULTS: Pearson, Spearman and Kendall rank coefficients showed a strong correlation (in all cases P < 0.001) between digital measurements of wounds against traditional hand-measured estimates. Intra-observer variation of wound length using digital elliptical measurement (DEM) gave a coefficient of variation of < 3.0%. Interobserver variation of wound length using DEM was < 6.5%. Variation from a standard known-size wound area was < 8.0% across 30 trials. CONCLUSIONS: This study shows a strong correlation between digital and traditional measurement techniques. The system can be easily deployed in routine clinical practice, providing an objective visual record, allowing remote in-depth analysis. PMID- 19125769 TI - Long-term review of driving potential following bilateral panretinal photocoagulation for proliferative diabetic retinopathy. AB - AIM: To determine the necessity for repeated Driver and Vehicle Licensing Agency (DVLA) visual field testing in people with diabetes who have had bilateral panretinal photocoagulation (PRP) for proliferative diabetic retinopathy. METHODS: A questionnaire survey was conducted of driving history in a cohort of people with diabetes who had been treated with bilateral PRP for proliferative retinopathy between 1988 and 1990. In addition, all similarly eligible subjects attending the diabetic retinal review clinic over a 12-month period who had had laser between 1991 and 2000 were questioned as to their driving status. RESULTS: Forty-five surviving patients from the 1988-1990 cohort were eligible and 25 returned the questionnaire (55%). Eight had never driven and 15 (13 with Type 1 diabetes) still held a valid licence, having passed the DVLA field test on a number of occasions. Neither of the two patients who had stopped driving reported failing the DVLA field test as the reason for stopping. All 12 of the patients directly questioned in the clinic were still driving and had passed at least one repeat DVLA test. CONCLUSIONS: People with Type 1 diabetes who have no further laser treatment for proliferative diabetic retinopathy can expect to retain their UK driving licence for at least 15 years following small-burn PRP, provided they maintain sufficient acuity. PMID- 19125770 TI - Biopsy of the posterior interosseous nerve: a low morbidity method for assessment of peripheral nerve disorders. AB - AIMS: The sural nerve is the commonest peripheral nerve biopsied to help in the diagnosis of peripheral neuropathy of unknown cause. However, associated complications limit its use. The aim was, as an alternative, to asses biopsy of the terminal branch of the posterior interosseous nerve (PIN) in the forearm. METHODS: PIN pathology was morphometrically quantified in 10 male patients with Type 2 diabetes and compared with six PIN biopsy specimens taken post mortem from male cadavers with no history of neuropathy or trauma. RESULTS: The PIN biopsy procedure provides a long (approximately 3 cm) mono- or bifascicular nerve biopsy with generous epineurial tissue and adjacent vessels. Our results show a significantly lower myelinated fibre density in subjects with diabetes [5782 (3332-9060)/mm(2)] compared with autopsy control material [9256 (6593 12,935)/mm(2), P < 0.007]. No postoperative discomfort or complications were encountered. CONCLUSIONS: A reduction in myelinated fibre density has previously been shown to be a clinically meaningful measure of neuropathy in diabetic patients. We demonstrate similar findings using the PIN biopsy. The PIN biopsy procedure fulfils the criteria for nerve biopsy and was well tolerated by the patients. It may be a possible alternative to sural nerve biopsy to allow for diagnosis of neuropathy. PMID- 19125771 TI - Neuroglycopenia and adrenergic responses to hypoglycaemia: insights from a local epidemic of serendipitous massive overdose of glibenclamide. AB - AIMS: Recently, an unlicensed aphrodisiac formulation originating from China known as 'Power 1 Walnut' penetrated the illicit markets of South East Asia including Singapore. Subsequent toxicological analyses revealed that each 'Power 1 Walnut' pill was illegally adulterated with two prescription drugs--sildenafil 1 mg (a PDE5 inhibitor) and glibenclamide 93-98 mg (a long-acting sulphonylurea). As the drug was peddled to numerous people, a local 'hypoglycaemia epidemic' ensued, of which a small cluster presented to our hospital with severe hypoglycaemia. The aim is to characterize the demographics, clinical and labouratory aspects and postulate mechanisms for the relatively atypical presentation. METHODS: A retrospective study of all the patients admitted between 13th January to 15th June 2008 with hypoglycaemia was done with acquisition of all relevant data after ethical approval from our DSRB. RESULTS: 15 patients (25 73 years old) presented with severe hypoglycaemia. All of them presented with neuroglycopenic symptoms (5-confusion, 6-drowsiness to loss of consciousness, 4 seizures and 1-coma) 12-36 h after ingestion of 'Power 1 Walnut'. Liquid chromatography-mass spectrometry (LC/MS) confirmed the presence of glibenclamide in the urine. None experienced a full-blown hypoglycaemia-associated autonomic response defined as sweating, sensation of warmth, anxiety, tremor, nausea, palpitations, tachycardia, and hunger. Only 4/16 patients had symptoms of a partial autonomic response, and 3/16 patients had tachycardia and/or hypertension. CONCLUSIONS: The above suggests an inappropriate autonomic and catecholamine response to severe hypoglycaemia in these cases of serendipitous glibenclamide overdose. Possible reasons for the blunted autonomic responses and apparent autonomic failure are reviewed. PMID- 19125772 TI - Comments on the use of bemiparin in diabetic foot ulcers. PMID- 19125773 TI - Insulin treatment in a patient with Type 2 diabetes and Stiff-person syndrome. PMID- 19125774 TI - Agenesis of the dorsal pancreas. PMID- 19125775 TI - Mutations in the third gene shown to alter fasting glucose levels in the population (G6PC2) are not a common cause of monogenic forms of pancreatic B-cell dysfunction. PMID- 19125776 TI - Sodium-glucose co-transporter-2 inhibitors: an emerging new class of oral antidiabetic drug. AB - The sodium-glucose co-transporter-2 (SGLT2) is a low-affinity transport system that is specifically expressed in the kidney and plays an important role in renal glucose reabsorption in the proximal tubule. Competitive inhibition of SGLT2 therefore represents an innovative therapeutic strategy for the treatment of hyperglycaemia and/or obesity in patients with type 1 or type 2 diabetes by enhancing glucose and energy loss through the urine. The observation that individuals with familial renal glycosuria maintain normal long-term kidney function provides some reassurance that this mode of action will not adversely affect renal function. Intense research in this therapeutic area has led to the discovery of novel SGLT2 inhibitors, each with different chemical, pharmacodynamic and pharmacokinetic profiles. This review outlines the biology, expression and pleotropic activity of the SGLT system and the pharmacological profile of SGLT2 inhibitors and provides a summary of preclinical and limited clinical data available to characterize the efficacy, safety and potential clinical utility of SGLT2 inhibitors in the management of diabetes. PMID- 19125777 TI - Fifty-two-week efficacy and safety of vildagliptin vs. glimepiride in patients with type 2 diabetes mellitus inadequately controlled on metformin monotherapy. AB - AIM: To examine the efficacy and safety of vildagliptin vs. glimepiride as add-on therapy to metformin in patients with type 2 diabetes mellitus in a 52-week interim analysis of a large, randomized, double-blind, multicentre study. The primary objective was to demonstrate non-inferiority of vildagliptin vs. glimepiride in glycosylated haemoglobin (HbA(1c)) reduction at week 52. METHODS: Patients inadequately controlled on metformin monotherapy (HbA(1c) 6.5-8.5%) and receiving a stable dose of metformin (mean dose 1898 mg/day; mean duration of use 36 months) were randomized 1:1 to receive vildagliptin (50 mg twice daily, n = 1396) or glimepiride (titrated up to 6 mg/day; mean dose 4.5 mg/day, n = 1393). RESULTS: Non-inferiority of vildagliptin was demonstrated (97.5% confidence interval 0.02%, 0.16%) with a mean (SE) change from baseline HbA(1c) (7.3% in both groups) to week 52 endpoint of -0.44% (0.02%) with vildagliptin and -0.53% (0.02%) with glimepiride. Although a similar proportion of patients reached a target HbA(1c) level of <7% with vildagliptin and glimepiride (54.1 and 55.5%, respectively), a greater proportion of patients reached this target without hypoglycaemia in the vildagliptin group (50.9 vs. 44.3%; p < 0.01). Fasting plasma glucose (FPG) reductions were comparable between groups (mean [SE] -1.01 [0.06] mmol/l and -1.14 [0.06] mmol/l respectively). Vildagliptin significantly reduced body weight relative to glimepiride (mean [SE] change from baseline -0.23 [0.11] kg; between-group difference -1.79 kg; p < 0.001) and resulted in a 10 fold lower incidence of hypoglycaemia than glimepiride (1.7 vs. 16.2% of patients presenting at least one hypoglycaemic event; 39 vs. 554 hypoglycaemic events, p < 0.01). No severe hypoglycaemia occurred with vildagliptin compared with 10 episodes with glimepiride (p < 0.01), and no patient in the vildagliptin group discontinued because of hypoglycaemia compared with 11 patients in the glimepiride group. The incidence of adverse events (AEs), serious AEs and adjudicated cardiovascular events was 74.5, 7.1 and 0.9%, respectively, in patients receiving vildagliptin, and 81.1, 9.5 and 1.6%, respectively, in patients receiving glimepiride. CONCLUSIONS: When metformin alone fails to maintain sufficient glycaemic control, the addition of vildagliptin provides comparable efficacy to that of glimepiride after 52 weeks and displays a favourable AE profile, with no weight gain and a significant reduction in hypoglycaemia compared with glimepiride. PMID- 19125779 TI - Endothelial cell/macrophage cocultures as a model to study Streptococcus suis induced inflammatory responses. AB - Endothelial cells and macrophages are thought to play a critical role in the inflammatory response that contributes to meningitis. To investigate the interactions between Streptococcus suis and these two cell types, we developed a coculture model composed of brain microvascular endothelial cells and macrophage like cells, and characterized the production of proinflammatory cytokines, chemokines, prostaglandin E(2) (PGE(2)), and matrix metalloproteinase 9 (MMP-9) following a challenge with bacteria. Streptococcus suis cells stimulated the secretion of all the inflammatory mediators as well as MMP-9 in the coculture model. Responses to S. suis infection were influenced by endothelial cell/macrophage ratios and were dependent on the multiplicity of infection. Except for IL-6, significantly higher amounts of inflammatory mediators and MMP-9 were produced with the coculture model at an endothelial cell/macrophage ratio of 1 : 10 than at a ratio of 1 : 1. When infected with S. suis, endothelial cells and macrophages acted in synergy to increase the secretion of IL-6 and PGE(2). Using a model that more closely reproduces the in vivo situation, we showed that S. suis can induce the secretion of high levels of inflammatory mediators and MMP 9, which may contribute to the development of meningitis. PMID- 19125780 TI - Retaining our nurses: why aren't we ahead of the pack? PMID- 19125781 TI - Factors associated with seclusion in a statewide forensic psychiatric service in Australia over a 2-year period. AB - Despite seclusion being described as one of the most ethically- and legally controversial management options available, it remains a widely-used clinical strategy for managing disruptive, aggressive, and violent behaviour. This study sought to determine how frequently seclusion was used, the common characteristics of those secluded and not secluded, and the degree to which the Level of Service Inventory - Revised: Screening Version (LSI-R: SV) could predict seclusion. The study was retrospective, covering the first 2 years of operation of a statewide forensic psychiatry hospital in Victoria, Australia. Data were collected from individual case files, electronic databases, and paper copies of records pertaining to violent incidents and episodes of seclusion. Eighty five (44%) of the 193 patients admitted during this period were secluded. Those secluded were significantly younger and had a more established psychiatric history. LSI-R: SV scores were significantly and positively associated with being secluded. A statistical model containing three LSI-R: SV items, along with age on admission and psychiatric history, achieved an area under the curve of 0.74. Seclusion is used on a regular basis in response to a range of different forms of aggressive behaviour of different severity. The LSI-R: SV demonstrated moderate-to-good accuracy in predicting seclusion and warrants further research using detailed prospective methodologies. PMID- 19125778 TI - Efficacy and safety of the dipeptidyl peptidase-4 inhibitor alogliptin in patients with type 2 diabetes inadequately controlled by glyburide monotherapy. AB - AIM: To evaluate the efficacy and safety of alogliptin, a potent and highly selective dipeptidyl peptidase-4 (DPP-4) inhibitor, in combination with glyburide in patients with type 2 diabetes inadequately controlled by sulphonylurea monotherapy. METHODS: After a 2-week screening period, adult patients 18-80 years of age entered a 4-week run-in/stabilization period in which they were switched from their own sulphonylurea medication to an equivalent dose of glyburide (open label) plus placebo (single blind). After the run-in period, patients were randomly assigned to double-blind treatment with alogliptin 12.5 mg (n = 203), alogliptin 25 mg (n = 198), or placebo (n = 99) for 26 weeks. The primary end point was change from baseline to week 26 in glycosylated haemoglobin (HbA1c). Secondary end-points included clinical response rates and changes in fasting plasma glucose, beta-cell function (fasting proinsulin, insulin, proinsulin/insulin ratio, and C-peptide, and homeostasis model assessment beta cell function), body weight, and safety end-points [adverse events (AEs), clinical laboratory tests, vital signs and electrocardiographic readings]. RESULTS: The study population had a mean age of 57 years and a mean disease duration of 8 years; it was well balanced for gender (52% women) and was mainly white (71%). The mean baseline HbA1c was approximately 8.1% in each group. Significantly greater least squares (LS) mean reductions in HbA1c were seen at week 26 with alogliptin 12.5 mg (-0.38%) and 25 mg (-0.52%) vs. placebo (+0.01%; p < 0.001), and more patients in the alogliptin 25-mg group had HbA1c levels < or =7.0% at week 26 (34.8%, p = 0.002) vs. placebo (18.2%). Proportionately more patients in the alogliptin 12.5 mg (47.3%) and 25 mg (50.5%) groups had an HbA1c reduction > or =0.5% from baseline compared with patients in the placebo group (26.3%; p < 0.001). Minor improvements in individual markers of beta-cell function were seen with alogliptin, but no significant treatment group differences were noted relative to placebo. Minor LS mean changes in body weight were noted across groups (placebo, -0.20 kg; alogliptin 12.5 mg, +0.60 kg; alogliptin 25 mg, +0.68 kg). AEs were reported for 63-64% of patients receiving alogliptin and 54% of patients receiving placebo. Few AEs were treatment limiting (2.0-2.5% across groups), and serious AEs (2.0-5.6%) were infrequent, similar across groups, and generally considered not related to treatment. The incidences of hypoglycaemia for placebo, alogliptin 12.5 mg and alogliptin 25 mg groups were 11.1, 15.8 and 9.6% respectively. CONCLUSIONS: In patients with type 2 diabetes inadequately controlled by glyburide monotherapy, the addition of alogliptin resulted in clinically significant reductions in HbA1c without increased incidence of hypoglycaemia. PMID- 19125782 TI - Ethical suicide research: a survey of researchers. AB - Research is needed to better understand and respond effectively to people who are suicidal. Involving people who are suicidal in research poses some ethical and pragmatic problems. The ethical problems and difficulties in obtaining approval to involve people who are suicidal in research has contributed to the current paucity of research that explores the suicidal experience. To explore some of these problems, a web-based survey of suicide researchers was undertaken. Researchers identified from published reports were contacted by email and invited to participate in a web-based survey. Researchers were asked to describe any problems they encountered, how ethical problems were negotiated or resolved, and any advice received from human research ethics committees. The main problems identified were accessing the population, maintaining confidentiality, the extent of care owed by the researcher to participants, and the facilitation of support to participants. As with clinical practice, ethical research involving people who are suicidal involves a process of sensitive engagement, and careful consideration and remediation of risk. PMID- 19125783 TI - Using an adapted reflecting team approach to learn about mental health and illness with general nursing students: an Australian example. AB - This paper describes how a reflecting team informed by outsider-witness practices was used in a classroom with a small group of Australian general nursing students to explore their understandings of mental illness and people with mental illness. The reflecting team process helped students to go beyond the media stereotypes of mental illness and the people who suffer from mental illness. It helped them develop new understandings of the lives of people who experience mental illness. The process also enabled students to learn more about stigma and its debilitating effects, to speak about it and to monitor their own language use, and to identify and explore their preferred interpersonal approaches in future practice settings when dealing with people showing signs of mental distress. PMID- 19125784 TI - Sociology and nursing: role performance in a psychiatric setting. AB - The role of sociology in nursing continues to cast new light on many aspects of health and illness. Over the last 20 years, nursing practice has seen sociological theory become a valuable clinical tool, both in the diagnosis and prognosis of a wide range of illnesses and long-term conditions. Nevertheless, of these, the sociological examination of mental health problems and its impact upon nursing practitioners has received little coverage, simply because, as a discipline, mental health nursing has historically been wedded to a biomedical model, one which continues to embrace psychiatry/psychology as the driving force in the diagnosis and treatment of psychopathology. Adopting a sociological approach, this paper brings to light previously unexplored insights into the way nurses interact with patients experiencing mental health problems. Drawing on social interactionist methodology, this paper considers depression and other mental health problems in relation to current psychiatric nursing practice. Specifically, the paper focuses on aspects of role performance and interpersonal care in a psychiatric setting, and the impact the individual role may have on the wider aspects of institutional and official practices. The paper concludes by making a number of recommendations/observations for nursing practice. PMID- 19125785 TI - Appreciating history: the Australian experience of direct-entry mental health nursing education in universities. AB - More than two decades since the introduction of comprehensive nursing education in Australia, the controversy regarding the type of undergraduate education that would best serve the needs of the mental health nursing profession continues. The ensuing debate tends to be based on a comparison between the current model of comprehensive education in the universities and the specialist mental health nursing programs that previously operated within the hospital system. The previous existence of a tertiary-based direct-entry mental health nursing program in Victoria is generally not recognized. The paper provides a brief overview of mental health nursing education from a historical perspective emphasizing the period following the commencement of the transfer of the nursing education. Articulating the Victorian experience of specialist undergraduate mental health nursing education within universities is essential as discussions about the most appropriate educational preparation for mental health nursing continues. PMID- 19125786 TI - Evaluation of a new mental health liaison role in a rural health centre in Rocky Mountain House, Alberta: a Canadian story. AB - This paper describes the evaluation of a mental health liaison (MHL) role in a rural community in Alberta, Canada. The role provides advocacy, education, indirect and direct client intervention, and follow up. It was developed to eliminate gaps in mental health care and build collaborative cultures between the local hospital, physicians' offices, mental health clinics, and community agencies. Obtaining stakeholder feedback was an important step in assessing initial service impact while providing directions for role refinement and future programme development. A total of 116 questionnaires were distributed to physicians, hospital staff, and community mental health assessing stakeholder perception relating to various functions of the MHL. A 50% (n = 58) response rate was achieved with broad representation from different partners, including 75% of local physicians. The majority of respondents positively perceived the roles, functions, and impact of the MHL, including relationship development across the hospital community, improved access to services, and perceived improved client outcomes. The results reinforced that the MHL service meets a previously unmet need in this rural setting. Findings are being used to refine roles, provide local learning and resource development, understand issues relating to programme development in other areas, and develop client level outcomes relating to the services delivered. PMID- 19125787 TI - Advocacy for mothers with psychiatric illness: a clinical perspective. AB - Motherhood is a challenging role and a life-changing experience. For women living with psychiatric illness, the challenge of motherhood is amplified. Psychiatric illness (including schizophrenia, affective and personality disorders) is associated with multiple adversities that can impair the capacity to parent. Social adversity, fluctuating symptoms, and medications and their related side effects, can create difficulties for the new mother as she adjusts to her role. The risk for relapse among women who are unwell is heightened during the post partum period. For many other women, the post-partum period is when psychiatric symptoms emerge for the first time. Equally important are the continuing concerns pertaining to infant well-being where maternal psychiatric illness is present. For mothers who exhibit symptoms at this time, a very real threat of protective removal exists. In the mother-infant setting, child protection legislation is biased towards the rights of the child. While there are cases for which this bias is clearly appropriate, there are less clear situations from which the infant is removed with little regard for the mother. Often mothers with psychiatric illness struggle to meet the cognitive, emotional, and financial demands of drawn-out custody proceedings. For these mothers, there is a paucity of appropriate support available, as will be evidenced throughout the present paper. There is an urgent need for professional advocacy to support women who are unwell in their transition to motherhood. The mental health nurse is able to fill a key advocating role in the perinatal psychiatric setting. Nurses in this role hold a unique position whereby social and community supports can be activated, while guidance is imparted from a ground-level standpoint. The nurse in this role has the capacity to liaise with authorities, negotiate service provision, and ensure that key parenting skills are acquired by the mother as she works to secure her role. Through the provision of proactive advocacy during this time, the nurse has the potential to ameliorate the outcomes of mothers who are unable to cope alone and the well-being of their infants. PMID- 19125788 TI - Foreign-born and Swedish-born families' perceptions of psychosis care. AB - The aim of the study was to describe how foreign-born and Swedish born families living in Sweden perceive psychosis care. Eleven foreign-born and 15 Swedish-born family members were interviewed and the data were analyzed using a phenomenographic approach. The findings showed three main descriptive categories: taking responsibility, access to care, and attitudes to psychosis. The degree of responsibility in the family decreased if there was easy access to care and support from health-care staff. Knowledge of psychosis was considered to be important in order to counteract prejudiced attitudes in the family and the community. Foreign-born families did not want to be treated differently from Swedes and stressed the importance of finding ways to communicate despite communication barriers. Foreign-born families also were affected by their experiences of psychiatric care and different beliefs about psychosis in their home country. The results indicate how important it is that health-care staff members treat families on equal terms. It is necessary to take the time to identify how to communicate in a good manner and to identify families' previous experiences of and beliefs about psychosis care in order to help families face prejudice in society and to see beyond the psychosis. PMID- 19125789 TI - Bachelor of Nursing students' beliefs about prognosis and outcomes for people with schizophrenia: a longitudinal study. AB - Many nursing students have unfavourable attitudes towards people with mental illness. This study aimed to examine if Australian Bachelor of Nursing students' beliefs about prognosis and long-term outcomes for people with schizophrenia changed through their course. The Attitudes and Beliefs about Mental Health Problems: Professional and Public Views questionnaire, which included a vignette about a person with schizophrenia, was used with a convenience sample of students. An interrupted time series approach to data collection was used, with data collected three times between 2005 and 2007. A university ethics committee approved the study. Data were analysed using SPSS version 15. Overall, the students recognized the person's need for professional help. By third year, students were more pessimistic about prognosis if the person did not receive professional help than in their earlier years of study. By third year, students also rated the long-term outcome to be 'creative or artistic' more favourably than in earlier years. The findings showed no significant differences in perceived overall negative and positive long-term outcomes across the three data collection points. The findings reinforce the need for theoretical and clinical education in mental health to be included and progressively increased in all years of study if attitudes are to be improved. PMID- 19125790 TI - Current state of diabetes care and a role for nurse practitioners. Introduction. PMID- 19125791 TI - The utility of FDG-PET in the preoperative staging of esophageal cancer. AB - Accurate staging of esophageal cancer is important when determining which patients will potentially benefit from curative surgery. The aim of this study was to evaluate the incremental effect of 2-fluoro-2-deoxyglucose positron emission tomography (FDG-PET) when used in addition to standard staging modalities. Patients referred to two surgeons in an Australian metropolitan teaching hospital with esophageal or esophago-gastric junction malignancy between May 2002 and December 2006 were included. Patients who had undergone prereferral treatment with chemotherapy or radiotherapy were excluded. Patients undergoing resection for gastrointestinal stromal tumors or high-grade dysplasia within Barrett's esophagus were also excluded. Clinical and non-clinical data were recorded prospectively. Pretreatment staging included routine CT scan and selective endoscopic ultrasound (EUS). FDG-PET was performed in patients judged to have curable disease on CT scanning and EUS. From a total of 130 eligible patients, 76 were judged to have curable disease on the basis of CT and EUS findings. Of these 76 patients, 19 (25%) were excluded from surgery due to additional information obtained from FDG-PET. The addition of FDG-PET to routine preoperative staging resulted in the exclusion from surgery of 19 (25%) patients who prior to the introduction of FDG-PET would have undergone attempted resection. FDG-PET should be performed in all patients under consideration for esophagogastric resection in order to avoid resection in patients with disseminated disease. PMID- 19125792 TI - An expression of squamous cell carcinoma antigen 2 in peripheral blood within the different stages of esophageal carcinogenesis. AB - The malignant transformation of esophageal mucosa is a progressive process, which includes basal cell hyperplasia, dysplasia, carcinoma in situ, and invasive esophageal squamous cell carcinoma (ESCC). The objectives of this study were to prove the relationship of squamous cell carcinoma antigen 2 (SCCA2) mRNA expression in peripheral blood with non-malignant lesion, premalignant lesion, and carcinoma of the esophagus at the same assay, as well as to evaluate whether or not SCCA2 mRNA expression in peripheral blood may be a biomarker for monitoring the premalignant lesion of the disease. The subjects consisted of 50 patients with basal cell hyperplasia, 50 patients with dysplasia, 50 patients with ESCC (12 carcinoma in situ, 38 carcinoma in invasive stage), and 50 controls who were pathologically diagnosed to be normal and whose esophageal mucosa were stained brown by iodine. All the subjects are residents of Feicheng, China, which is considered an area with a high incidence of esophageal cancer. All subjects were diagnosed by two separate histopathologists, and the expression of SCCA2 mRNA in peripheral blood was detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Furthermore, SCCA2 concentration in the serum was measured using an enzyme-linked immunosorbent assay (ELISA). In the cancer group, SCCA2 mRNA expression was also detected in 20 tissues of esophageal cancer. By using the band intensity ratios of SCCA2 to beta-actin, with a positive cut-off value of > or = 0.4, the positive rates of the SCCA2 mRNA expression in peripheral blood were found to be 82% (41/50), 60% (30/50), 48% (24/50), and 36% (18/50) in the cancer, dysplasia, basal cell hyperplasia, and control groups, respectively. The positive rate of the cancer group was significantly different from the three other groups (P < 0.05), and there was also a significant difference in the SCCA2 mRNA expression between the dysplasia group and the control group (chi(2)=5.769, P= 0.016). In the multinomial logistic regression analysis, the odds ratios (ORs) were 1.71 [95% confidence interval (95% CI), 0.73-3.99] in the basal cell hyperplasia group, 2.77 (95% CI, 1.14 6.71) in the dysplasia group, and 7.87 (95% CI, 2.88-21.55) in the cancer group after being adjusted for age, gender, smoking index, drinking index, and family history of esophageal cancer. The SCCA2 mRNA expression in peripheral blood was then divided into different grades according to the band intensity ratios of SCCA2 to beta-actin. By using a positive cut-off value of > or = 0.4, the testing sensitivities in the basal cell hyperplasia, dysplasia, and cancer groups were found to be 48%, 60%, and 82%, respectively, with the same testing specificity at 64%. On the other hand, SCCA2 mRNA expression in peripheral blood had a 97.5% agreement with that in tissue, and there was a significant correlation between the ELISA SCCA2 levels in the serum and the SCCA2 mRNA expression levels in the peripheral blood (r= 0.80, P= 0.01). The results indicate that SCCA2 mRNA expression in peripheral blood is linked with the different stages of esophageal pathological changes, despite the fact that SCCA2 mRNA was not a biomarker for screening early esophageal cancer. This knowledge may be useful in monitoring the processes of change that occur in esophageal premalignant lesions among subjects who live in a high-incidence area. PMID- 19125793 TI - Cortactin overexpression in the esophageal squamous cell carcinoma and its involvement in the carcinogenesis. AB - The aim of this study is to examine whether dysregulated expression of cortactin occurs in esophageal squamous cell carcinoma (ESCC) and is involved in the development of ESCCs. An immunohistochemistry study for cortactin expression was performed on 46 pairs of surgically resected non-tumor and ESCC tumor tissues and murine tumors of esophagi induced by a carcinogen. The results show increased cortactin expression in 20 and in 22 to a lesser extent, out of a total 46 ESCC tumor tissues. Increased cortactin was also detected in the premalignant lesions, the early stage dysplasia and carcinoma in situ, of ESCC tumor tissues. Differential polymerase chain reaction results showed slight increases in the EMS1 gene only in two of 10 ESCC tumor tissues, suggesting that EMS1 gene amplification is not the only mechanism for cortactin overexpression. In the mouse model induced by treatment with 4-nitroquinoline 1-oxide and arecoline, increased cortactin was detected in the epithelia with hyperkeratosis, papillomas, and ESCCs with invasion into the submucosa, respectively. Overall, we observed cortactin overexpression in early and late stages of human ESCCs and carcinogen-induced murine ESCCs, suggesting a role for cortactin in esophageal carcinogenesis. PMID- 19125794 TI - PTEN polymorphisms and the risk of esophageal carcinoma and gastric cardiac carcinoma in a high incidence region of China. AB - PTEN, as a tumor suppressor gene, plays an important role in regulating cell growth, proliferation, and apoptosis. Two common polymorphisms, -9C/G and IVS4 ( /+), may alter susceptibility to the disease. To test the hypothesis that the genetic variations of PTEN play a role in the etiology of esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA), a population based case-control study was conducted in 350 ESCC patients, 257 GCA patients, and 634 healthy controls from a high-incidence region of Hebei province, China. The PTEN polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP). The results showed that the family history of upper gastrointestinal cancer (UGIC) significantly increased the risk of developing ESCC and GCA (the age, gender and smoking status adjusted OR = 1.73 and 1.67; 95% CI = 1.29-2.32 and 1.28-2.19, respectively). The overall distribution of the PTEN -9C/G genotype was not significantly different between cancer patients and controls. Compared with the PTEN IVS4-/- genotype, the IVS4+/+ genotype significantly decreased the risk of ESCC and GCA development, the adjusted OR was 0.64 (95% CI = 0.44-0.94) and 0.63 (95% CI = 0.41-0.98), respectively. Stratification analysis by gender, age, smoking status and family history of UGIC showed that the PTEN IVS4-/+ genotype only reduced the risk of ESCC (adjusted OR = 0.55, 95%CI = 0.34-0.90) among subjects with family history of UGIC. While the IVS4+/+ genotype decreased the susceptibility to both ESCC and GCA (adjusted OR = 0.61 and 0.57, 95% CI = 0.37-0.98 and 0.34-0.98, respectively) among male subjects, the IVS4+/+ genotype only decreased the risk of ESCC development among subjects younger than 55 years (adjusted OR = 0.43, 95% CI = 0.21-0.85). In addition, the haplotype analysis found that the -9C/IVS4- haplotype increased the risk of developing ESCC and GCA (OR = 1.31 and 1.24, 95% CI = 1.08-1.58 and 1.001-1.53). Our results suggested that the PTEN IVS4+/+ homozygote may play a protective role in the development of ESCC and GCA, while the haplotype -9C/IVS4- might be the risk factor of the development of ESCC and GCA in the high incidence region population of Hebei province, China. PMID- 19125795 TI - Psychiatric illness delays diagnosis of esophageal cancer. AB - Evidence suggests that patients with psychiatric illnesses may be more likely to experience a delay in diagnosis of coexisting cancer. The association between psychiatric illness and timely diagnosis and survival in patients with esophageal cancer has not been studied. The specific aim of this retrospective cohort study was to determine the impact of coexisting psychiatric illness on time to diagnosis, disease stage and survival in patients with esophageal cancer. All patients with a diagnosis of esophageal cancer between 1989 and 2003 at the Portland Veteran's Administration hospital were identified by ICD-9 code. One hundred and sixty patients were identified: 52 patients had one or more DSM-IV diagnoses, and 108 patients had no DSM-IV diagnosis. Electronic charts were reviewed beginning from the first recorded encounter for all patients and clinical and demographic data were collected. The association between psychiatric illness and time to diagnosis of esophageal cancer and survival was studied using Cox proportional hazard models. Groups were similar in age, ethnicity, body mass index, and history of tobacco and alcohol use. Psychiatric illness was associated with delayed diagnosis (median time from alarm symptoms to diagnosis 90 days vs. 35 days in patients with and without psychiatric illness, respectively, P < 0.001) and the presence of advanced disease at the time of diagnosis (37% vs. 18% of patients with and without psychiatric illness, respectively, P= 0.009). In multivariate analysis, psychiatric illness and depression were independent predictors for delayed diagnosis (hazard ratios 0.605 and 0.622, respectively, hazard ratio < 1 indicating longer time to diagnosis). Dementia was an independent risk factor for worse survival (hazard ratio 2.984). Finally, psychiatric illness was associated with a decreased likelihood of receiving surgical therapy. Psychiatric illness is a risk factor for delayed diagnosis, a diagnosis of advanced cancer, and a lower likelihood of receiving surgical therapy in patients with esophageal cancer. Dementia is associated with worse survival in these patients. These findings emphasize the importance of prompt evaluation of foregut symptoms in patients with psychiatric illness. PMID- 19125796 TI - A change in clinical practice: a partially stapled cervical esophagogastric anastomosis reduces morbidity and improves functional outcome after esophagectomy for cancer. AB - Cervical esophagogastric anastomoses are commonly used for reconstruction after esophagectomy because of the lower mortality rate associated with an anastomotic leak compared to intrathoracic anastomoses. However, cervical esophagogastric anastomoses have been criticized for their higher leak rates, stricture formation and greater need for later dilatations when compared with intrathoracic anastomoses. Multiple studies have looked at varying techniques to improve the outcome of the cervical esophagogastric anastomosis. This study was performed to determine whether a partially stapled (posterior stapled wall and anterior hand sewn wall) anastomosis reliably reduced leaks and the need for later dilatation. From January 2001 to March 2006, 168 patients who underwent cervical esophagogastric anastomosis following esophagectomy (transhiatal or three-hole) for cancer were identified. Beginning in September 2003, the partially stapled technique was introduced and used in 79 patients. Clinical outcomes were compared to patients in whom hand-sewn technique was used (n = 89). Outcomes related to anastomotic leak, other hospital complications, length of stay, postoperative dilatations and survival were compared using Student's t-tests and chi-square tests (P < 0.05), as well as multiple regression analyses. An anastomotic leak occurred in 10 (12.7%) patients who received a partially stapled anastomosis. A hand-sewn anastomosis was complicated by an anastomotic leak in 24 patients (27.0%). This difference was statistically significant (P = 0.021). This lowered incidence of leak was associated with an earlier initiation of oral feeds (median 7 vs. 9.5 days, P < 0.001) and a reduction in hospital stay (median 10 vs. 15 days, P < 0.001). Furthermore, dysphagia associated with stricture requiring postoperative dilatations was markedly diminished in the stapled anastomosis [23 (31.3%) vs. 49 (55.1%), P = 0.001]. The partially stapled cervical esophagogastric anastomosis significantly decreased the incidence of postoperative anastomotic leaks and the need for postoperative dilatation to treat strictures compared to the hand-sewn anastomosis. PMID- 19125797 TI - A single institutional non-randomized retrospective comparison between definitive chemoradiotherapy and radical surgery in 82 Japanese patients with resectable esophageal squamous cell carcinoma. AB - This retrospective study was conducted to compare the treatment results between radical surgery and definitive chemoradiotherapy for resectable squamous cell carcinoma of the esophagus. Between June 2000 and May 2005, 82 consecutive patients were selected for this study in which 33 were treated with chemoradiotherapy and 49 with surgery. The patients in the chemoradiotherapy (CRT) group received 2-4 cycles of 5-fluorouracil (1000 mg/m(2)/day, day 1-4, continuous) combined with cisplatin (75 mg/m(2), day 1, bolus) plus 50.4 Gy of radiation, while those in the surgery group were treated by an esophagectomy with radical node dissection. Eighteen surgical patients received postoperative chemotherapy. The baseline clinical TNM stage was similar between the two groups. With a median follow-up period of 36 months (range: 23-84 months) with 47 survivors (57%), the 3-year overall survival rates (P = 0.22) and disease-free survival rates (P = 0.16) were 48% and 44% in the chemoradiotherapy group versus 65% and 59% in the surgery group, and lacked statistical significance. This non randomized study on patients with resectable squamous cell carcinoma of the esophagus showed that chemoradiotherapy could result in survival comparable with conventional surgery in spite of selection bias of patients. There is a trend toward improved survival with surgery versus definitive CRT. PMID- 19125798 TI - Preoperative versus postoperative radiotherapy for locally advanced gastroesophageal junction and proximal gastric cancers: a comparison of normal tissue radiation doses. AB - Locoregional relapse occurs in over half of gastric cancer patients who undergo potentially curative resection. Adjuvant chemoradiation reduces locoregional relapse, but often requires irradiating large fields and is limited by poor patient tolerance. This study explores the potential dosimetric benefit in reducing the radiation dose to normal structures by treating gastroesophageal (GE) junction/proximal gastric cancers with preoperative rather than adjuvant radiotherapy. Five cases of GE junction/proximal gastric cancer patients treated postoperatively with curative intent were selected. The actual target contours were then modified to reflect hypothetical target volumes which would have been used had the patients been treated preoperatively. Hypothetical preoperative treatment plans were generated for each patient based on these modified contours. The hypothetical preoperative treatment plans were then compared to the actual postoperative plans with respect to dose-volume parameters including lung mean dose, lung V20, heart V20 and V30, and mean doses to abdominal structures. Target volumes were smaller with preoperative treatment, with an average reduction of 23%. Comparative dose-volume histogram (DVH) analysis showed the resultant composite lung doses were reduced in the preoperative plans by 50-79%. In all patients, the proportion of lungs receiving at least 20 Gy (V20) was substantially reduced using preoperative treatment (1.9% vs. 9.7% in the 3-D conformal patient; mean of 3.1% vs. 17.6% in the intensity modulated radiation therapy patients). Likewise, the volume of heart receiving at least 30 Gy was dramatically reduced in all preoperative plans (15.8% vs. 35.4%). Doses to the kidneys, liver and spinal cord were comparable in both approaches. Preoperative treatment of GE junction and proximal gastric cancer patients offers the potential to decrease the radiation dose received by normal thoracic structures. PMID- 19125799 TI - Symptoms experienced during 24-h pH monitoring and their relationship to outcome after laparoscopic total fundoplication. AB - Some patients having a 24-h pH monitoring test prior to laparoscopic fundoplication experience no symptoms at all in spite of having a positive test, and other patients experience only atypical symptoms in spite of having a positive test. This study investigates the postoperative outcome of such patients. All patients underwent esophageal manometry and 24-h esophageal pH monitoring before laparoscopic total fundoplication. Patients were divided into three groups based on their symptom profile recorded during a positive 24-h pH monitoring: those with typical symptoms (n = 104), those with atypical symptoms (n = 28) and those who experienced no symptoms at all (n = 23). The outcomes measured were heartburn score (0-10), dysphagia composite score (0-45) and satisfaction score (0-10) at 12 months after surgery. Outcome analysis reveals the heartburn scores were significantly reduced postoperatively for all groups of patients. At 1 year after surgery, there was no difference among the three groups of patients in terms of heartburn score and dysphagia composite scores, nor the experience of bloating, belching, or their willingness to repeat surgery. Despite one group experiencing no symptoms, and another group atypical symptoms during a positive pH study, the postoperative satisfaction scores for these two groups was good, but significantly less (P = 0.03, P = 0.02, respectively) than the group of patients with a typical symptom index. In conclusion, patients who experience only atypical symptoms or no symptoms at all during their preoperative positive 24-h pH monitoring may still obtain a good result from antireflux surgery. However, these symptom profiles should alert the surgeon that such patients may have an outcome which is not as good as patients who experience only typical symptoms during a pH study. PMID- 19125800 TI - The inaccuracy of ICD-9-CM Code 530.2 for identifying patients with Barrett's esophagus. AB - Barrett's esophagus (BE) is a chronic complication associated with gastroesophageal reflux disease. The ICD-9-CM code used for BE, 530.2, is also used for patients with 'ulcer of the esophagus.' We aimed to determine if the ICD 9-CM code of 530.2 is reliable for identifying cases of Barrett's esophagus within databases for research purposes. We reviewed the records of all patients assigned code 530.2 at two university medical center hospitals and a veterans' administration hospital over a cumulative 16-year period. Billing records provided information about where the code was assigned, whether it was a major or minor diagnosis, and if the code was assigned on multiple occasions for each patient. Histology and endoscopy records were reviewed to confirm the diagnosis of Barrett's esophagus. Among 435 patients with code 530.2 in their records, 354 (82%) had an esophageal biopsy reported and 393 (90%) had an endoscopy report available for review. Only 182 (42%) had specialized intestinal metaplasia documented in a biopsy from an area of salmon-colored mucosa arising above the esophagogastric junction (51% of those with histology available). There were 288 patients (66%) with an endoscopic diagnosis of Barrett's esophagus (73% of those with an endoscopy reported). Variables associated with documented specialized intestinal metaplasia were age > or = 60 (OR 2.3; 95% CI 1.4-3.7), multiple assignments of 530.2 (OR 3.2; 95% CI 2.0-5.0), and assignment of 530.2 in a gastrointestinal (GI) clinic or an endoscopy unit (OR 3.5; 95% CI 2.0-6.3). The positive predictive value of the code being assigned in a GI location was 48% (95% CI 43-54%). Therefore, ICD-9-CM code of 530.2 is not specific for the diagnosis of Barrett's esophagus. The usage of code 530.2 in a GI setting was not sufficiently predictive of BE to be reliable for rigorous epidemiological studies. PMID- 19125801 TI - Gastroesophageal reflux disease and non-small cell lung cancer. Results of a pilot study. AB - The sharp rise in the frequency of adenocarcinoma and relative decrease of squamous cell carcinoma of the respiratory and digestive systems, raises suspicion of a common element in their carcinogenetic cascade, which could result in similar trends in cell-type distribution changes of esophageal and lung cancers. The possible role of chemical irritation caused by gastroesophageal reflux disease (GERD) in non-small cell lung cancer (NSCLC) patients was investigated. There was no significant difference between the adenocarcinoma and the squamous cell carcinoma groups, neither in the composite DeMeester scores nor in any of the separate parameters of the complex score investigated. However, the ratio of detected gastroesophageal reflux cases was considerably higher than in the average population. This factor may be one element of a multifactorial cancer promotion. PMID- 19125802 TI - Esophageal replacement using cryopreserved tracheal graft. AB - Esophageal replacement using artificial material is not a new concept. Prior experiences with prostheses, allografts and composite grafts have not proved particularly successful. The aim of this study is to investigate whether cryopreserved animal trachea is suitable for the replacement of the esophagus. In 12 beagle dogs a 6-cm-long segment of the cervical esophagus was removed, and was replaced with cryopreserved trachea, which had been stored for 21 days on -86 degrees C. The proximal and distal ends of the esophagus were joined together with the graft by simple continuous suture (Biosyn 3/0) and covered with the sternohyoid flap. Postoperatively 16 hematological parameters were measured. The dogs were planned to be euthanized at random on days 28, 42 and 56 after the operation. Tests for air leak were performed and the inner diameter of the graft was measured to detect shrinkage. The microscopic structure of the graft was analyzed using haematoxylin and eosin staining. There was no indication of insufficiency. Based on the air leak test the sutures withheld properly. The inner diameter of the graft narrowed from an average 19 mm (+/- 1 mm) to 15.8 mm (+/- 0.6 mm). In length, the graft shortened from an average 60 mm to 47 mm (+/- 3 mm). No feeding difficulty was observed. In two cases wound suppuration was found involving only the cutaneous and subcutaneous layers. Concerning the laboratory parameters, only the fibrinogen level and white blood cell count showed temporary although significant changes. Histology findings on the 56th day showed absolute integration of the trachea with the esophagus, with disintegration of the tracheal cartilages. Cryopreserved trachea seems to be suitable for the replacement of a 5-6-cm-long esophageal segment. PMID- 19125803 TI - Recurrent cutaneous myoepithelioma treated using Mohs micrographic surgery: case report and review of the literature. PMID- 19125804 TI - In vitro two-dimensional echocardiographic imaging of a stented porcine bioprosthetic valve: the bent strut artifact. AB - BACKGROUND: Echocardiographic imaging of a stented valve bioprosthesis can reveal apparent inward deflection of one or more struts. It could be assumed that this finding is related to actual strut distortion as opposed to an artifact of off axis imaging. OBJECTIVE: To determine whether normal (nondistorted) bioprosthetic struts can appear by artifact to be bent inward on two-dimensional echocardiographic imaging. METHODS: A production-quality porcine bioprosthetic aortic valve was imaged in vitro using standard two-dimensional echocardiographic techniques. Apparent strut distortion on echocardiographic imaging was investigated relative to prosthesis orientation to the transducer. RESULTS: The appearance of inward strut distortion was produced when two of three struts were simultaneously imaged, including imaging in an off-axis long axis orientation and from above or below the prosthesis. CONCLUSION: Apparent inward distortion of bioprosthetic struts can be simulated in vitro using a normal, nondistorted valve, and is common if two struts are simultaneously imaged. A finding of inward distortion of strut tips on in vivo imaging should be used with caution, since the finding may not be representative of actual strut anatomy. PMID- 19125805 TI - Detection of severe stenosis and total occlusion in the left anterior descending coronary artery with transthoracic Doppler echocardiography in the emergency room. AB - BACKGROUND: The noninvasive measurement of coronary flow velocity in the left anterior descending artery (LAD) has recently been realized by using the transthoracic Doppler echocardiography (TTDE). A couple of investigations demonstrated that the diastolic-to-systolic peak velocity ratio (DSVR) by TTDE is a simple and noninvasive method for the detection of severe stenosis in the elective settings. However, the usefulness of DSVR by TTDE in the emergency settings has not been evaluated. OBJECTIVE: The purpose of this study was to assess the clinical feasibility to document the LAD flow by TTDE in emergency patients who complained of chest pain. METHODS: We studied 49 consecutive patients with acute coronary syndrome who were going to undergo emergency coronary angiography (CAG) for the anatomical diagnosis and the facilitated percutaneous coronary intervention (PCI). Prior to CAG, we recorded the LAD flow by TTDE and measured the diastolic peak velocity (DVp), systolic peak velocity (SVp), and their ratio, DSVR (DVp/SVp) of LAD flow. RESULTS: By CAG, the culprit lesions actually resided in the proximal LAD in 36 patients. Among the 36 patients, we detected the Doppler LAD flow in 29. Five out of 7 patients who were unable to detect the LAD flow revealed total occlusions by CAG. DSVR of the LAD is significantly lower in 17 patients who showed severe stenoses (>90%) than those in the rest of 12 patients who did not show such critical stenoses (1.44 +/ 0.16 vs 2.10 +/- 0.26, P < 0.0001). CONCLUSION: In the emergency settings, a noninvasive assessment of the LAD flow by TTDE accurately estimates the critical stenotic lesions of the LAD. PMID- 19125806 TI - Clinical significance of positive isovolumetric relaxation velocity of pulsed wave tissue Doppler imaging. AB - OBJECTIVE: Among the pulsed-wave tissue Doppler imaging (pw-TDI) parameters, there are two different pw-TDI velocities (IVRa and IVRb) after systolic velocity, but before Ea velocity. In our study, we investigated the clinical importance of these two velocities in left ventricular diastolic dysfunction (LVDDF) evaluation. METHODS: One hundred and eighty cases without exclusion criteria were included in the study. Cases with a transmitral E to A flow (E/A) ratio below 1 were assigned to group 2. In cases with an E/A ratio between 1 and 2, the pw-TDI parameters were taken into consideration. Cases with an Ea/Aa ratio above 1 were assigned to group 1 and cases with an Ea/Aa ratio 1 or below than 1 were assigned to group 3. Group 1 (n: 68) represented normal diastolic left ventricular (LV) inflow while group 2 (n = 87) represented impaired relaxation and group 3 (n = 25) represented pseudonormal LV inflow. RESULTS: In our study, we found that IVRa velocity was lower in group 1 compared to group 2 and group 3 (P < 0.001 and P = 0.038, respectively). Similarly, this velocity was significantly different in group 3 and group 2 such as it was higher in group 2 compared to group 3 (P = 0.022). There was no difference in IVRb velocity and IVRa/IVRb ratio among the groups. A negative correlation was found between IVRa velocity and Ea velocity (r = 44%, P < 0.001). Positive correlation was found between IVRa velocity and isovolumetric relaxation time (r = 18%, P = 0.014) and also between IVRa velocity and Aa velocity (r = 19%; P = 0.010). CONCLUSION: Based on the results of our study, we concluded that IVRa velocity is an important pw-TDI parameter in the evaluation of LVDDF, especially in differentiating pseudonormal LVDDF type from normal LV inflow. PMID- 19125807 TI - The role of Tissue Doppler imaging in the noninvasive detection of chronic rejection after heterotopic cardiac transplantation in rats. AB - BACKGROUND: Chronic rejection is a risk factor for the development of cardiac allograft vasculopathy (CAV) in heart transplant recipients. A useful animal model to study the role of immunosuppressive strategies in the prevention of chronic rejection involves heterotopic abdominal cardiac transplantation in rats. The detection of rejection and concurrent CAV traditionally involves subjective serial palpation of the graft from a scale of 0 to 4, with 4 indicating vigorous beats. Recent advances in murine echocardiography, in particular Tissue Doppler imaging (TDI), may allow for objective in vivo monitoring of chronic rejection in this transplant model. OBJECTIVE: The objective of this study was to compare the diagnostic accuracy of murine echocardiography as compared to the abdominal palpation heart score for the noninvasive detection of chronic cardiac graft rejection. METHODS: In an animal model of heterotopic cardiac transplantation, 18 male Fischer and Lewis rats were used as donors and recipients, respectively. Abdominal palpation and murine transthoracic echocardiography were performed to assess in vivo function of the transplanted heart. Left ventricular (LV) structure and function and TDI indices, including endocardial velocity (Vendo) and strain rate (SR), were evaluated in the ectopic heart. Graft tissues were processed for histological examination and graded for chronic rejection. RESULTS: Abdominal palpation scores were obtained in all 18 rats; score 1 (n = 5); score 2 (n = 4); score 3 (n = 6); and score 4 (n = 3). The mean LV ejection fraction was significantly (P <0.01) lower in score 3 and 4 grafts as compared to score 1 grafts. There was no correlation between the abdominal palpation score and LV systolic function. There was a significant relationship between decreasing Vendo or SR values and increasing grades of rejection (r = 0.65, P <0.05 and r = 0.75, P < 0.05, respectively). CONCLUSION: TDI of the transplanted heart in rats is feasible, reproducible, and more sensitive than palpation for the detection of chronic rejection. PMID- 19125808 TI - Myocardial contrast echocardiography in assessment of stable coronary artery disease at intermediate dobutamine-induced stress level. AB - BACKGROUND: Myocardial contrast stress echocardiography (stress MCE) is a novel method for diagnosing coronary artery disease (CAD). Few studies have compared the diagnosis of ischemia by stress MCE to angiographic CAD. METHODS: Dobutamine stress MCE and SonoVue contrast infusion were performed before an elective percutaneous coronary intervention in 37 patients (8 women) aged 45-75 years with symptomatic CAD and at least one significant coronary artery stenosis measured by quantitative coronary angiography (QCA). The total and regional perfusion and wall motion (WM) were scored as normal or abnormal and attributed to the three main epicardial coronary arteries using a 17-segment left ventricular model. RESULTS: An intermediate stress level was obtained in 29 (78%) patients, and 2 (5%) patients obtained peak stress. A perfusion defect was detected in 92% and WM abnormality in 57% of the patients at peak stress (P < 0.01). By perfusion, 70% of stenoses were both detected and correctly anatomically located, compared to 42% by WM (P < 0.01). All 21 patients with multivessel disease and/or proximal left anterior descending (LAD) stenosis measured by QCA were identified by stress induced perfusion defects, while only 11 of them were identified by WM abnormalities (P < 0.01). CONCLUSION: Perfusion scoring is superior to WM scoring during stress MCE for diagnosing significant CAD in patients obtaining intermediate stress level, in particular, when multivessel disease or proximal LAD stenosis is present. PMID- 19125809 TI - Combined assessment of myocardial perfusion and diastolic function enhances risk stratification in patients with anterior wall myocardial infarction. AB - OBJECTIVE: Evaluate the utility of a combined risk stratification scheme including diastolic dysfunction and "no-reflow," to identify high-risk patients following acute myocardial infarction (AMI). BACKGROUND: Recent studies have demonstrated that the "no-reflow" phenomenon (defined by myocardial contrast echocardiography) and severe diastolic dysfunction (identified by Doppler echocardiography) identify patients at high risk for mortality following AMI. METHODS: We evaluated 111 patients with recent anterior acute myocardial infarction from July 2000 to June 2004. Diastolic function and myocardial perfusion was evaluated by echocardiography. Patients were placed into 1 of 3 groups based on diastolic function and myocardial perfusion: Group 1 (normal perfusion and normal diastolic function), Group 2 (abnormal perfusion or abnormal diastolic function), and Group 3 (abnormal perfusion and abnormal diastolic function). We compared the long term all-cause mortality within these groups. RESULTS: Patients in each group were similar with respect to myocardial infarction size as defined by biomarkers, extent and severity of coronary artery disease, and medical and interventional therapy. Mortality was much higher in Group 3 (26.9%) compared to Group 1 (0%) and Group 2 (15.2%) (p = 0.048). CONCLUSION: Combined assessment of diastolic function and myocardial perfusion enhances risk stratification post myocardial infarction. PMID- 19125810 TI - The use of 3D contrast-enhanced CT reconstructions to project images of vascular rings and coarctation of the aorta. AB - BACKGROUND: Aortic arch and pulmonary artery anomalies make up a group of vascular structures that have complex three-dimensional (3D) shapes. Tortuosity as well as hypoplasia or atresia of segments of the aortic arch or pulmonary artery makes the conventional two-dimensional (2D) imaging difficult. METHODS: Nine patients with native coarctation or recoarctation and 4 patients with a vascular ring had a CT scan as a part of their clinical evaluation. There were 7 males. The mean age was 11.7 years. (range 19 days to 29 years) The mean weight was 22.7 kg (range 3.3-139.0 kg). The dicom data from contrast CT scans were converted by the Amira software package into a 3D image. The areas of interest were selected. The images were then projected in 3D on a standard video monitor and could be rotated 360 degrees in any dimension. RESULTS: Adequate CT scans and 3D reconstructions were obtained in 12 of 13 patients. There were 85-1,044 slices obtained in the adequate studies. We could not reconstruct a 3D image from a patient's CT scan that had only 22 slices. The anatomy defined by 3D was compared to 2D CT imaging and confirmed by cardiac catheterization or direct visualization in the operating room in the 12 patients with adequate 3D reconstructions. In 5 of 12 patients, 3D reconstructions provided valuable spatial information not observed in the conventional 2D scans. CONCLUSION: We believe that 3D reconstruction of contrast-enhanced CT scans of these complex structures provides additional valuable information that is helpful in the decision-making process. PMID- 19125811 TI - The role of myocardial contrast echocardiography to assess the origin of a mass in right cardiac cavities. AB - We report the case of a patient with hepatocellular carcinoma who was admitted to our hospital with fatigue and edema of lower extremities. Transthoracic echocardiographic examination revealed a mobile echogenic cavoatrial mass that infiltrated the inferior vena cava and extended along the vessel protruding into the right cardiac cavities. The differential diagnosis included a tumor mass originating from the liver and subsequently infiltrating the inferior vena cava and extending into the right cardiac cavities or a large thrombus formed on the tumor mass that infiltrated the inferior vena cava. PMID- 19125812 TI - Death caused by splenic artery aneurysm rupture during dobutamine-atropine stress echocardiography: case report and literature review. AB - A 55-year-old man developed hypertension and acute epigastric pain during dobutamine-atropine stress echocardiography (DASE). Evaluation-including a helical computed tomography (CT) scan of the abdomen and pelvis, as well as surgical exploration-revealed a ruptured splenic artery aneurysm. The patient died, despite multiple surgical interventions and a massive blood product transfusion. Impressively, no deaths from DASE have been previously reported. Additionally, no adverse sequelae during DASE have been reported in patients with an unruptured abdominal aortic aneurysm >or=4 cm in diameter or with an unruptured intracranial aneurysm. We report the first case, to our knowledge, of death caused by splenic artery aneurysm rupture during DASE. Splenic artery aneurysm rupture during DASE, though rare, can lead to death. PMID- 19125813 TI - Renal artery fibromuscular dysplasia is a cause of refractory hypertension in the elderly. AB - Fibromuscular dysplasia (FMD) is predominantly a disease of younger women, but it can occur and cause refractory hypertension in the elderly. We present here classic angiographic and intravascular ultrasound images of FMD in a 70-year-old woman with refractory hypertension. Renal artery FMD should be included in the differential diagnosis of refractory hypertension even in older patients since recognizing and treating this condition will add favorably to the outcome of these patients. PMID- 19125815 TI - Liver receptor homolog-1 localization in the nuclear body is regulated by sumoylation and cAMP signaling in rat granulosa cells. AB - Liver receptor homolog-1 (LRH-1; NR5A2) is an orphan member of the nuclear receptor superfamily, mainly expressed in endoderm-derived tissues and in the ovary. In ovarian granulosa and luteal cells, LRH-1 regulates the expression of genes associated with ovarian steroidogenesis. LRH-1 can be transported to transcriptionally inactive nuclear bodies after conjugation with small ubiquitin related modifier (SUMO). In the present study, we investigated the effects of SUMO modification at five lysine residues of LRH-1 in rat granulosa cells. Lysine 289 could be conjugated with SUMO-1 in vitro, and the mutation K289R increased transcriptional activity of LRH-1, suggesting that SUMO conjugation is associated with transcription repression. Coexpression of SUMO-1 targets LRH-1 to the dot like nuclear bodies, but the effect of lysine mutations on blocking subnuclear localization depended on the cell type. In COS-7 cells, mutation of either K173 or K289 prevented SUMO-1-mediated translocation of LRH-1 into nuclear bodies and also reduced the conjugation by SUMO-1, suggesting that K289 and K173 are two important sites involved in SUMO-1 modification. In granulosa cells, three or more altered lysine residues were required for nucleoplasm retention. This result suggests that multiple lysine residues are targets for SUMO conjugation in vivo and granulosa cells are more sensitive to SUMO-1-mediated LRH-1 localization to nuclear bodies. Nuclear body localization of LRH-1 was suppressed by forskolin and cholera toxin. Forskolin treatment obviously influences the expression of members involved in the SUMO pathway. The results obtained in the present study suggest that cAMP signaling could change the dynamic process of sumoylation and repress LRH-1 targeting to nuclear speckles in rat granulosa cells. PMID- 19125817 TI - Unique glycine-activated riboswitch linked to glycine-serine auxotrophy in SAR11. AB - The genome sequence of the marine bacterium 'Candidatus Pelagibacter ubique' and subsequent analyses have shown that while it has a genome as small as many obligate parasites, it nonetheless possesses a metabolic repertoire that allows it to grow as one of the most successful free-living cells in the ocean. An early report based on metabolic reconstruction indicated that SAR11 cells are prototrophs for all amino acids. However, here we report experimental evidence that 'Cand. P. ubique' is effectively auxotrophic for glycine and serine. With glucose and acetate added to seawater to supply organic carbon, the addition of 125 nM to 1.5 microM glycine to growth medium containing all other nutrients in excess resulted in a linear increase in maximum cell density from 1.14 x 10(6) cells ml(-1) to 8.16 x 10(6) cells ml(-1) (R(2) = 0.992). Serine was capable of substituting for glycine at 1.5 microM. 'Cand. P. ubique' contains a glycine activated riboswitch preceding malate synthase, an unusual genomic context that is conserved in the SAR11 group. Malate synthase plays a critical role in central metabolism by enabling TCA intermediates to be regenerated through the glyoxylate cycle. In vitro analysis of this riboswitch indicated that it responds solely to glycine but not close structural analogues, such as glycine betaine, malate, glyoxylate, glycolate, alanine, serine or threonine. We conclude that 'Cand. P. ubique' is therefore a glycine-serine auxotroph that appears to use intracellular glycine level to regulate its use of carbon for biosynthesis and energy. Comparative genomics and metagenomics indicate that these conclusions may hold throughout much of the SAR11 clade. PMID- 19125818 TI - Apoptosis as a post-phagocytic winnowing mechanism in a coral-dinoflagellate mutualism. AB - This study was aimed at detecting apoptosis as a post-phagocytic mechanism of symbiont selection during the onset of symbiosis in larvae of the scleractinian coral Fungia scutaria. Larvae were infected with one of three Symbiodinium types: freshly isolated homologous ITS-type C1f from adult F. scutaria, heterologous C31 from adult Montipora capitata, known to be unable to successfully colonize F. scutaria larvae, and type B1 from the symbiotic sea anemone Aiptasia spp. Apoptosis was detected by the activation of caspases, enzymes specific to apoptosis. Caspase activity was measured in situ by cleavage of a specific fluorophore and detection with confocal microscopy. At 6 h post infection, there was a significant increase in caspase activation in gastrodermal cells in C31 infected larvae, compared with larvae infected with C1f or B1 types. Compared with control larvae infected with C31, which had decreased infection rates present by 24 h post infection, when C31-infected larvae were incubated with a broad-scale caspase inhibitor, the per cent of larvae infected with C31 did not significantly decrease over time. This indicates that the reduction in infection success observed in untreated C31-infected larvae can be rescued with inhibition of caspases and apoptosis. This suggests the presence of a post-phagocytic recognition mechanism. Larvae infected with freshly isolated B1 retained infection success over time compared with C31-infected larvae, suggesting that there is host discrimination between heterologous algae. Initiation of this post phagocytic response may occur more readily with a highly specific heterologous symbiont type such as C31, compared with a generalist heterologous type such as clade B1. PMID- 19125816 TI - Insights on Escherichia coli biofilm formation and inhibition from whole transcriptome profiling. AB - Biofilms transform independent cells into specialized cell communities. Here are presented some insights into biofilm formation ascertained with the best characterized strain, Escherichia coli. Investigations of biofilm formation and inhibition with this strain using whole-transcriptome profiling coupled to phenotypic assays, in vivo DNA binding studies and isogenic mutants have led to discoveries related to the role of stress, to the role of intra- and interspecies cell signalling, to the impact of the environment on cell signalling, to biofilm inhibition by manipulating cell signalling, to the role of toxin/antitoxin genes in biofilm formation, and to the role of small RNAs on biofilm formation and dispersal. Hence, E. coli is an excellent resource for determining paradigms in biofilm formation and biofilm inhibition. PMID- 19125820 TI - Embracing the power and challenge of change. PMID- 19125821 TI - Emergency nurse practitioners: an underestimated addition to the emergency care team. PMID- 19125822 TI - Performance assessment and simulation fidelity for dummies. PMID- 19125823 TI - Review article: Animal bites: an update for management with a focus on infections. AB - Animal bites are a significant public health problem, with an estimated 2% of the population bitten each year. The majority of bites are from dogs and risk factors include young children, men, certain dog breeds and unrestrained dogs. The risk of infection following bites differs among animal species and is dependent on animal dentition and oral flora. Recent studies have demonstrated a broad range of pathogens isolated from infected bite wounds, with Pasteurella species being the predominant isolate from dog and cat bite wounds. Controversy exists about the use of prophylactic antibiotics; however, they are currently only recommended for high-risk bite wounds. Two fatal cases of Australian bat lyssavirus have been reported and bats are the only identified reservoir in Australia. All bat bites are of high risk and should receive post-exposure prophylaxis for rabies. Workers handling bats should be offered routine immunization. PMID- 19125824 TI - Interrater reliability of the Australasian Triage Scale for mental health patients. AB - OBJECTIVE: To evaluate interrater reliability of the Australasian Triage Scale (ATS) for mental health patients in ED. METHODS: In a prospective descriptive study, triage nurses were shown video vignettes of simulated scenarios of mental health presentations. Trieurs (raters) were asked to allocate an ATS category (rating) to each case. The primary outcome was the degree of interrater reliability for each simulated case. Also assessed were differences between raters or settings, grouped by level of ED activity, state of origin, hospital type and familiarity with appropriate guidelines. Chi-squared analysis was used for independent categorical variables; the Friedman test was used to compare the triage scores between busy and quiet ED scenarios. Ordinal data results were compared using opartchi. RESULTS: All 90 eligible participants were enrolled. The highest interrater concordance was 65.6% whereas the lowest interrater concordance was 53.3%. Significant association occurred between the distribution of triage ratings, ED activity level and the state of origin. A busy ED resulted in the assignment of a more urgent ATS category and decrease in concordance. CONCLUSIONS: There is a need to develop and implement a validated, standardized national triage tool for mental health patients. The ATS per se is insufficient to ensure acceptable interrater reliability, particularly during busy periods in the ED, and between states. Given the influence the ATS has on key outcomes, it is imperative for this tool to be robust. PMID- 19125825 TI - Validation of a verbal dyspnoea rating scale in the emergency department. AB - OBJECTIVE: Use of a verbal dyspnoea rating scale in the emergency department (ED) has many potential benefits, providing information to clinicians otherwise not afforded by objective parameters. In the present study, we aimed to investigate the validity of a verbal dyspnoea rating scale, previously validated in the setting of cardiac stress tests, among patients presenting to the ED with acute shortness of breath (SOB). METHODS: This was a prospective observational study conducted at an inner-urban adult tertiary hospital. A convenience sample of patients presenting with SOB to the ED had objective data collected at triage and 30 min later, including respiratory rate (RR), oxygen saturation (S(a)O(2)), heart rate (HR) and systolic blood pressure (SPB). These were correlated with the participants' subjective response to the question: 'On a scale from 0 to 10, how bad is your SOB, with zero being no SOB and 10 the worst SOB you could ever imagine?' Spearman correlations were then calculated between objective and subjective breathlessness measures. RESULTS: For 253 breathless ED patients (mean age 60.6 years, 126 male), verbal dyspnoea scores at triage correlated with RR (r = 0.77, P < 0.001), S(a)O(2) (r =-0.43, P < 0.001), HR (r = 0.35, P < 0.001) and SPB (r = 0.19, P < 0.05). Thirty minutes later, correlations remained significant for RR (r = 0.74, P < 0.001), S(a)O(2) (r =-0.39, P < 0.001) and HR (r = 0.40, P < 0.001). CONCLUSION: A verbal numerical SOB rating scale is a valid measure of breathlessness in the ED, and might therefore provide useful insight into a symptom that is otherwise unmeasurable. PMID- 19125826 TI - Pain score documentation and analgesia: a comparison of children and adults with appendicitis. AB - OBJECTIVE: There is a growing body of evidence suggesting that administration of analgesia in paediatric ED is inadequate. The present study was designed to assess pain score documentation and provision of opioid analgesia to children and adults with confirmed appendicitis in a mixed Australian ED. METHOD: A retrospective chart review of all adults and children with histologically confirmed appendicitis diagnosed in the Townsville ED during 2006 was performed. Data collected included pain score documentation, weight, opioid dose, oral analgesia, time of presentation, level of doctor and prehospital analgesia. RESULTS: Data were collected for 106 adults and 39 children. Among them, 13 (33%) children compared with 79 (75%) adults had a pain score documented (OR 0.16, 95% CI 0.07-0.37, P < 0.001). And 11 (28%) children compared with 79 (75%) adults received i.v. morphine (OR 0.13, 95% CI 0.06-0.31, P < 0.001). Administration of oral analgesia lowered the likelihood and pain score documentation increased the likelihood of receiving morphine in both children and adults. CONCLUSION: Documentation of pain scores and provision of i.v. morphine is generally poor. Children are less likely than adults to have a pain score documented, or receive i.v. morphine when presenting with appendicitis. PMID- 19125827 TI - Improving documentation of endotracheal intubation in an adult emergency department. AB - OBJECTIVE: To evaluate the effect of an educational intervention and documentary pro forma on the adequacy of documentation of intubation in an adult ED. METHODS: A structured medical record review was performed before and after a multi-pronged educational programme and introduction of a specifically designed guide to documentation. Records were assessed for adequacy of documentation by the presence or absence of predetermined elements. Analysis focused on five aspects considered to be most important for future clinical care: drugs and doses used, grade of view, size of endotracheal tube, confirmation of placement and adverse events/difficulties encountered. RESULTS: Sixty-one and sixty-eight charts were included in the pre-intervention and post-intervention groups, respectively. The drugs and doses used were documented in 92% and 93%. The endotracheal tube size was recorded 82% and 91% of the time. Grade of laryngoscopy was documented in 35% and 46%. Confirmation of endotracheal tube placement was 69% and 84%. Presence or absence of adverse events was recorded in 37% and 54%. All five elements were present in 8.2% and 25% of medical records. CONCLUSION: Documentation improved slightly following the intervention, but was still unsatisfactory. We believe that to achieve an adequate level of documentation in the medical record for an episode of intubation, there needs to be a formal and structured mechanism, either via mandatory use of a specifically designed form and/or by participation in an organized data registry. PMID- 19125828 TI - Helicopter use in rural trauma. AB - OBJECTIVE: To profile a helicopter emergency medical service in rural Australia. To assess patient injury severities and outcomes. To compare missions involving ambulance officers with physicians. To determine any time advantage of the aircraft over ground transfer. METHODS: Intention-to-treat analysis using retrospective case note review of all helicopter emergency medical service trauma patients from January 2004 to November 2006. Global positioning system mapping technology was used to compare one-way road transfer times with two-way helicopter retrieval. RESULTS: Two hundred and twenty-two missions were identified from the helicopter log. Forty missions were aborted in flight. Of 182 patients transported, 11 records were incomplete, leaving 171 for analysis. Fifty (29%) patients transported had an Injury Severity Score (ISS) > 15; the average ISS was 12.30 (standard error of the mean 0.82). The average calculated distance flown was 160.4 nautical miles (standard error of the mean 5.29; range 28-360 nautical miles). There was no significant difference in ISS between ambulance officers and physician groups (t = -1.17, P = 0.25, 95% CI -7.37-1.91). There was no difference in the incidence of severe injury (ISS > 15, P = 0.39) or mortality (P = 0.33) when the groups were compared. Air transport was significantly faster beyond 100 km, with a mean difference of 48 min (P = 0.00). CONCLUSION: We could not identify a significant survival benefit attributable to the addition of a doctor, although numbers for this comparison were small. Predicting missions where flight physicians might provide benefit remain imprecise and should be a priority area for prospective evaluation. We have demonstrated that in the absence of special circumstances, a helicopter response within 100 km from base does not improve time to definitive care. PMID- 19125829 TI - External injury documentation in major trauma victims is inadequate: grounds for routine photography in the emergency department? AB - OBJECTIVE: There is no widely accepted measure of clinical documentation quality in the ED. The present study creates a measure for comparing the quality of clinical documentation of external injuries with autopsy reports. This is used to discuss the advantages and disadvantages of introducing routine photography to improve clinical documentation of injuries. METHODS: This retrospective case series addressed all non-surviving major trauma patients (Injury Severity Score > or =15) presenting to St. Vincent's Hospital ED, Sydney, within the 5 year period from 1 July 2002 to 30 June 2007. Comparison between clinical and autopsy documentation of external injuries was completed for each major trauma patient. RESULTS: Of the 48 major trauma patients, there were an average of 11.6 injuries missed in documentation per patient (P < 0.001, 95% CI 8.6-14.6). ED documentation recorded on average 29% (95% CI 26%-32%) of the external injuries that appeared in the autopsy report. We call this percentage the external injury documentation rate. The external injury documentation rate was influenced by injury count and body region, but was not influenced by age, sex, severity (using the Abbreviated Injury Scale and Injury Severity Score), or whether the clinician used a trauma survey or standard progress notes or not, and there was no visible trend over time. CONCLUSION: Clinical documentation of external injuries in major trauma is poor. This is presumably because of many factors, including time pressures and high-stress environments. A possible strategy to improve this documentation is routine photography, which should offer both clinical and legal benefits. PMID- 19125830 TI - Comparison of high- and low-fidelity mannequins for clinical performance assessment. AB - OBJECTIVE: A pilot study exploring the differences between high- and low-fidelity mannequins in the assessment of clinical performance. METHODS: Standardized clinical scenarios were used to test 12 intensive care paramedics (ICP). Each ICP was randomly assigned to a scenario using either a high-fidelity (SimMan) or low fidelity mannequin (Laerdal Heart Start 2000), followed by a crossover assessment using the alternative scenario. We examined both the objective and subjective outcomes. Objective performance was assessed by three independent assessors, all accredited Advanced Paediatric Life Support instructors. Subjective outcomes were measured by assessment questionnaires and a rating scale. RESULTS: The overall proportion that passed the high-fidelity mannequin scenario was 0.47 compared with 0.58 in the low-fidelity mannequin scenario. The difference was -0.11 (95% CI -0.32-0.11). The subjective outcomes were charted and presented within the article. The ICP preferred the use of high-fidelity mannequin for assessment purpose. CONCLUSION: There was no significant objective difference between the two mannequins. PMID- 19125831 TI - Comparative review of the clinical use of intranasal fentanyl versus morphine in a paediatric emergency department. AB - OBJECTIVES: Comparison of intranasal fentanyl (INF) and parenteral morphine in children in an ED. Primary objective was to compare time to analgesia from presentation, with secondary objectives to assess patient profiles, specifics of opiate analgesics used plus rate of i.v. access for analgesia alone. SETTING: Tertiary paediatric ED. METHODS: Retrospective review of case notes identified through controlled drug register. Patients who received INF and/or parenteral morphine between 1 January and 31 March 2005 (before introduction of fentanyl) and in corresponding months in 2006 and 2007 were included. RESULTS: A total of 617 patients were included. Geometric mean time to analgesia was statistically different for INF versus morphine in 2006 (31.2 min, SD 2.6 vs 55.6 min, SD 2.4) and in 2007 (23.7 min, SD 2.8 vs 53.1 min, SD 3.1) (both P < 0.000). Mean initial dose of INF in 2007 was 1.46 mg/kg (SD 0.11) compared with 1.32 mg/kg (SD 0.36) in 2006. Mean total dose in 2007 was 2.14 mg/kg (SD 0.93), increased from 1.60 mg/kg (SD 0.56) in 2006. INF was used most commonly for fractures and morphine for abdominal pain. The i.v. access for opiate analgesia decreased from 161/161 (100%) in 2005 to 99/237 (41.8%) in 2007. CONCLUSION: Use of INF in our paediatric ED setting was associated with a significantly reduced time to analgesia for patients requiring immediate analgesia compared with parenteral morphine. Since the introduction of an INF protocol to our department in mid 2005, INF use has increased, with a corresponding decrease in the use of morphine and a reduction in i.v. access for analgesia. PMID- 19125832 TI - Gamma-hydroxybutyrate: a 30 month emergency department review. AB - OBJECTIVES: To describe epidemiology, symptomatology, resource use and complications in patients attending the ED following gamma-hydroxybutyrate (GHB) ingestion. METHODS: Retrospective chart review of GHB-related emergency attendances over 30 months. RESULTS: One hundred and seventy emergency attendances attributed to GHB ingestion occurred. Monthly attendance rate doubled during the study, and was highest on public holidays and weekends between 04.00 and 08.00 hours. The majority (63%, 95% CI 55.7-70.3) were young men (median 22 years). GHB was ingested alone in 62 cases (36%, 95% CI 29.6-43.9). Poly substance ingestion was common (108 cases; 64%; 95% CI 56.1-70.4). The commonest presenting symptom was altered conscious state (89%, 95% CI 84.1-93.5) with 54% (95% CI 46.6-61.6) having low Glasgow Coma Score (GCS 3-8) on arrival at the ED. Eight per cent (95% CI 3.6-11.6) were intubated. Eighty-seven per cent (95% CI 79.8-93.8) with low GCS were not intubated. There were no serious adverse outcomes or fatalities. Recovery time from ED arrival to high GCS (9-15) was rapid (median 76 min, interquartile range 80). Overall median length of stay was 199 min (interquartile range 162). CONCLUSIONS: This is the largest GHB-related case series to date. Attendance rate doubled during the study, and peaked at times of lowest staffing. Poly-substance ingestion is common. Attendances are of high acuity with decreased conscious state and airway threat. With close conservative management, most recover quickly without adverse sequelae. PMID- 19125833 TI - Survey of the availability of lipid emulsion infusion in Australasian emergency departments. PMID- 19125835 TI - Are cortical tubers epileptogenic? Evidence from electrocorticography. AB - The purpose of this study was to characterize the epileptogenicity of tubers and surrounding cortex in patients with tuberous sclerosis complex (TSC). Three pediatric patients with TSC and intractable epilepsy underwent surgical resection of tubers associated with epileptogenic foci. In all patients, presurgical imaging revealed a prominent tuber that correlated on electroencephalography (EEG) with frequent interictal epileptiform discharges and electrographic seizures. Intracranial electrocorticography (ECoG) was performed using subdural grids placed over the tuber and surrounding cortex and depth electrodes positioned directly within the tuber. In all three patients, the depth electrode within the tuber was electrographically silent, whereas the surrounding cortical tissue showed significant epileptiform activity. The tuber and the electrically active adjacent cortex were resected. The patients experienced a drastic reduction in seizure frequency postsurgery. Epileptogenicity of cortical tubers may derive not from the lesion itself, but rather from the perturbation or abnormal development of the surrounding cortex. PMID- 19125836 TI - Voxel-based 3D MRI analysis for the detection of epileptogenic lesions at single subject level. PMID- 19125837 TI - P-gp expression in epilepsy. PMID- 19125838 TI - Sleep-related tongue biting may not be a sign of epilepsy: a case of sleep related faciomandibular myoclonus. PMID- 19125839 TI - Rapid onset of florid psychotic symptoms during video-EEG telemetry: undetected complex partial status? PMID- 19125840 TI - Italian League Against Epilepsy (LICE). Introduction. PMID- 19125841 TI - Recommendations for the management of "febrile seizures": Ad Hoc Task Force of LICE Guidelines Commission. AB - Febrile seizures are the most common seizure disorder in childhood, affecting 2 5% of children. Simple febrile seizure is defined as a short (<15 min) generalized seizure, not recurring within 24 h, that occurs during a febrile illness not resulting from an acute disease of the nervous system in a child aged between 6 months and 5 years, with no neurologic deficits and no previous afebrile seizures. These recommendations address the instructions for management of the first febrile seizures, giving criteria for hospital admission, diagnosis, differential diagnosis, and treatment of a prolonged seizure. The authors stressed the benign prognosis of the majority of cases and the risk factors for recurrence of febrile seizures and appearance of epilepsy later on. Both continuous and intermittent anticonvulsant therapy are efficacious in preventing single febrile seizures, but side effects may be so important to overcome the benefits. These treatments are indicated in very selected patients. PMID- 19125842 TI - Italian consensus conference on epilepsy and pregnancy, labor and puerperium. AB - To facilitate an integrated and rational approach to the care of women with epilepsy of childbearing potential, a group of experts appointed by Italian scientific societies in the fields of epileptology, neonatology, pediatrics, neuropediatrics, child neuropsychiatry, obstetrics, and gynecology held a joint meeting in Santa Trada di Cannitello, Reggio Calabria, Italy, on October 15-16, 2004, with the aim of reaching consensus on the optimal management of these women. An ad hoc system for the classification of available published evidence and the opinions of experts was developed and used to grade recommendations on different aspects related to counseling, diagnostic, and treatment issues. The present document summarizes available evidence on the reciprocal interactions between epilepsy, antiepileptic drugs, fertility, contraception, pregnancy, delivery, breastfeeding, and the offspring. Recommendations are made concerning the information and counseling that should be provided to women with epilepsy with respect to issues related to contraception, conception, pregnancy, labour, and puerperium. More detailed recommendations on the same issues are provided to physicians and other healthcare professionals involved in the care of these women, with special reference to choice of effective contraception, optimization of antiepileptic drug therapy, use of prenatal diagnostic tests and other monitoring procedures, and appropriate management practices in relation to childbirth, puerperium, and the care of the child. PMID- 19125843 TI - Epilepsy and postpartum depression. AB - PURPOSE: Postpartum depression occurs in 10-20% of women who have recently given birth, but less than half of cases are recognized. The purpose of this study was to examine whether there was a difference in the rate of postpartum depression between patients with epilepsy and healthy controls, and to determine the relationship between epilepsy and postpartum depression. METHODS: The sample consisted of 55 patients with epilepsy and 55 patients without epilepsy. All patients were recruited at the Regional Epilepsy Center and at the Department of Obstetrics and Gynecology, University of Milan, St. Paolo Hospital (Italy). Every woman was assessed with the Edinburgh Postnatal Depression Scale and a clinical interview to screen for postpartum depression. RESULTS: We found a statistically significant higher rate of postpartum depression in patients with epilepsy compared with controls (p < 0.05). DISCUSSION: These findings highlight the importance of screening for postpartum depression; it can improve the recognition of the disorder and can provide adequate treatment and follow-up. PMID- 19125844 TI - Changes in sex steroid levels in women with epilepsy on treatment: relationship with antiepileptic therapies and seizure frequency. AB - PURPOSE: Reproductive dysfunction in epilepsy is attributed to the seizures themselves and also to antiepileptic drugs (AEDs), which affect steroid production, binding, and metabolism. In turn, neuroactive steroids may influence neuronal excitability. A previous study in this cohort of consecutive women with epilepsy showed that patients with more frequent seizures had higher cortisol and lower dehydroepiandrosterone sulfate levels than those with rare or absent seizures. The present study was aimed at evaluating, in these same women, the possible relationship between some clinical parameters, seizure frequency, AED therapies, and sex hormone levels. METHODS: Estradiol (E2), progesterone (Pg), sex hormone-binding globulin (SHBG), and free estrogen index (FEI) were measured during the luteal phase in 113 consecutive females, 16-47 years old, with different epilepsy syndromes on enzyme-inducing AED (EIAED) and/or non-enzyme inducing AED (NEIAED) treatments, and in 30 age-matched healthy women. Hormonal data were correlated with clinical parameters (age, epilepsy syndrome, disease onset, and duration), seizure frequency assessed on the basis of a seizure frequency score (SFS), and AED therapies. RESULTS: E2, Pg, and FEI were lower, whereas SHBG levels were higher in the epilepsy patients than in the controls. However, sex steroid and SHBG levels were not different between groups of patients categorized according to SFS. Therapies with EIAEDs accounted for changes in E2 levels and FEI. CONCLUSIONS: Despite globally decreased sex steroid levels in serum, actual hormone titers were not significantly correlated with SFS in consecutive epilepsy women; rather, these hormonal changes were explained by AED treatments, mainly when EIAED polytherapies were given. PMID- 19125845 TI - Hyperhomocysteinemia in patients with epilepsy: does it play a role in the pathogenesis of brain atrophy? A preliminary report. AB - PURPOSE: Brain atrophy (BA) is observed in 20-50% of patients with epilepsy. Hyper-total-homocysteinemia (hyper-tHcy), which occurs in 10-40% of patients, is considered to be a risk factor for cardiovascular diseases and BA. The present study was aimed at investigating the possible association of hyper-tHcy with BA in a population of patients with epilepsy. METHODS: Fifty-eight patients (33 M/25 F, 43.5 +/- 13.1 years of age) chronically treated with antiepileptic drugs (AEDs) and 60 controls matched for age and sex were enrolled. All participants underwent determination of plasma tHcy, folate, vitamin B(12), and C677T methylene-tetrahydrofolate-reductase (MTHFR) polymorphism genotyping, and brain magnetic resonance imaging (MRI). RESULTS: Patients exhibited significantly higher tHcy and lower folate levels than controls; hyper-tHcy was significantly associated with the variables group (patients vs. controls), MTHFR genotype, and their interaction terms. BA was observed in 30.1% of patients and was significantly associated with hyper-tHcy (beta = 0.45, p = 0.003) and polytherapy (beta = 0.31, p < 0.001). DISCUSSION: Our investigation suggests that hyper-tHcy plays a role in the development of BA in patients with epilepsy. Although the real origin of this phenomenon is not yet fully elucidated, experimental data support the hypothesis of a link of the neuronal Hcy-mediated damage with oxidative stress and excitotoxicity. PMID- 19125846 TI - Incidence of epilepsy in extremely low-birthweight infants (<1,000 g): a population study of central and southern Sardinia. AB - PURPOSE: With the development of intensive care, the survival of extremely low birthweight (ELBW) infants (<1,000 g) has greatly improved. The aim of our study was to report the incidence of epilepsy after a follow-up of >7 years in a population of ELBW children, born in central and southern Sardinia between 1991 and 2000. METHODS: We analyzed data of 104 children. All infants had had serial cranial ultrasound echography (CUE) in the neonatal period and some also had magnetic resonance imaging (MRI). At last follow-up we evaluated the occurrence of epilepsy through a review of clinical charts and a structured telephone interview. RESULTS: In 11 (10.6%) of 104 of children we observed febrile seizures (FS). Epilepsy occurred in 9 (8.6%) of 104 ELBW children, and in these patients a frequent positive family history for epilepsy and/or FS was present. In four epilepsy patients CUE was highly pathologic, showing intraventricular hemorrhage (IVH) of grade IV and in two mildly abnormal (IVH of grade I-II). In three additional children with normal neonatal ultrasound scan, a later magnetic resonance imaging (MRI) study revealed lesions related to neonatal insult. DISCUSSION: In our ELBW population, epilepsy had an incidence clearly superior to that expected in infancy (8.6% vs. 0.6-0.8%). A frequent positive familiar history for epilepsy and/or FS suggests that a genetic predisposition may also play a role. Subjects with highly abnormal CUE are a subgroup with high risk for seizures; however, epilepsy can occur even with normal CUE. PMID- 19125847 TI - Prognostic factors in patients with mesial temporal lobe epilepsy. AB - PURPOSE: To disclose clinical, electrophysiologic, and neuroradiologic factors correlated to prognosis in patients with mesial temporal lobe epilepsy (MTLE). METHODS: One hundred thirty-six MTLE patients were studied for family history, clinical characteristics, instrumental data [electroencephalography (EEG), video EEG, neuroimaging], and outcome. The population was divided into drug-resistant (DR: 108 patients, 79.4%) and non-drug-resistant (NDR: 28 patients, 20.6%) groups; all variables were analyzed in the two groups. RESULTS: The comparison between the two groups shows a relation between resistance to therapy and febrile seizures (FS) (DR 43.5% vs. NDR 17.8%, p = 0.008), mesial temporal sclerosis (MTS) (DR 64.8% vs. NDR 32.1%, p = 0.0025), early age at seizure onset (DR 23.1% vs. NDR 3.6% p = 0.0160), and epileptiform interictal abnormalities (DR 89.7% vs. NDR 68%, p = 0.010). FS were more frequent in patients with MTS than in patients without (46.28% vs. 26.3%, p = 0.0199). Sixty-nine patients underwent surgery and 85.3% of them had a good outcome. CONCLUSION: MTLE is a heterogeneous syndrome. Establishing the factors responsible for and associated with drug resistance is important for therapeutic purposes, as prompt diagnosis of drug resistance must lead to early surgical management. This study shows that FS, MTS, early age at seizure onset, and epileptiform interictal abnormalities are negative prognostic factors and that FS are related to MTS. PMID- 19125848 TI - Diffusion-weighted magnetic resonance imaging in patients with partial status epilepticus. AB - PURPOSE: Diffusion-weighted magnetic resonance imaging (DWI) is used to detect changes in the distribution of water molecules in regions affected by various pathologies. Like other conditions, ictal epileptic activity, such as status epilepticus (SE), can cause regional vasogenic/cytotoxic edema that reflects hemodynamic and metabolic changes. This study describes the electroclinical and neuroimaging findings in 10 patients with partial SE whose DWI evaluation disclosed periictal changes related to sustained epileptic activity. PATIENTS AND METHODS: In this retrospective study we selected 10 patients with partial SE of different etiologies (six acute symptomatic SE; four with previous epilepsy and concomitant precipitating factors) who underwent video-EEG (electroencephalography) monitoring and a DWI study during the periictal phase. We analyzed ictal electroclinical features and DWI changes in the acute phase and during the follow-up period. RESULTS: DWI images revealed significant signal alterations in different brain regions depending on the location of ictal activity. DWI changes were highly concordant with the electroclinical findings in all 10 patients. As the SE resolved and the clinical conditions improved, DWI follow-up showed that the signal alterations gradually disappeared, thereby documenting their close relationship with ictal activity. CONCLUSIONS: This study confirms the usefulness of DWI imaging in clinical practice for a more accurate definition of the hemodynamic/metabolic changes occurring during sustained epileptic activity. PMID- 19125849 TI - Neurosteroids and epileptogenesis in the pilocarpine model: evidence for a relationship between P450scc induction and length of the latent period. AB - PURPOSE: Cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) catalyzes the initial step in the biosynthesis of neurosteroids within the brain. We sought to determine which cells express P450cc and whether neurosteroids play a role in the regulation of epileptogenesis following pilocarpine-induced status epilepticus (SE). METHODS: Rats experienced uninterrupted SE or SE terminated with diazepam at 60, 120, and 180 min. P450scc induction in CA3 hippocampus was determined by double immunolabeling with P450scc antiserum and monoclonal antibodies against GFAP (astrocytes), RIP (oligodendrocytes), or heme oxygenase-1 (microglia). RESULTS: SE was associated with P450scc induction in many astrocytes and a small number of microglia and oligodendrocytes in the hippocampal CA3 strata radiatum and lacunosum-moleculare. The extent of P450scc induction increased with increasing SE duration. Paradoxically, increased P450scc induction in rats experiencing SE for 180 min or more was associated with the delayed onset of spontaneous recurrent seizures. Treatment with the 5 alpha-reductase inhibitor finasteride (100 mg/kg/day for 25 days), which inhibits the synthesis of gamma aminobutyric acid (GABA)(A) receptor modulating neurosteroids such as allopregnanolone, was associated with a significant reduction in time to the onset of spontaneous seizures in rats exposed to 180-min but not 90-min SE. DISCUSSION: P450scc is induced by SE in a diverse population of hippocampal glia. Induction of P450scc is associated with the delayed onset of spontaneous seizures. Conversely, inhibition of neurosteroid synthesis accelerated the onset of spontaneous seizures, but only in animals exhibiting significant increases in P450scc. These findings suggest that induction of neurosteroid synthesis in reactive glial cells is associated with delayed onset of spontaneously recurrent seizures. PMID- 19125850 TI - Lack of alpha 1b-adrenergic receptor protects against epileptic seizures. AB - PURPOSE: The role of alpha 1b-adrenergic receptor (alpha 1b-AR) in relation with neuronal degeneration, drug addiction, and seizure susceptibility has recently emerged. In particular, mice that overexpress alpha 1b-AR undergo spontaneous epileptic seizures and progressive neuronal loss in a variety of brain areas. Therefore, one should expect that the blockade of alpha 1b-AR leads to anticonvulsant and neuroprotective effects. However, the lack of alpha 1b-AR antagonists does not allow testing of this hypothesis. METHODS: The development of alpha 1b-AR knockout (KO) mice led us to measure seizure susceptibility and neurodegeneration following systemic excitotoxins in these mice. RESULTS: We found that alpha 1b-AR KO mice are markedly resistant to kainate- and pilocarpine induced seizures. Moreover, when marked seizure duration and severity are obtained by doubling the dose of chemoconvulsants in alpha 1b-AR KO, neuronal degeneration never occurs. CONCLUSIONS: These data indicate that alpha 1b-AR per se plays a fundamental role in the mechanisms responsible for seizure onset, severity, and duration, whereas the brain damage observed in alpha 1b-AR overexpressing mice is likely to be a secondary phenomenon. In fact, the absence of alpha 1b-AR confers resistance to neurotoxicity induced by seizures/chemoconvulsants. These data, although confirming a pivotal role of alpha 1b-AR in modulating seizure threshold and neuronal death, offer a novel target, which may be used to develop novel anticonvulsants and neuroprotective agents. PMID- 19125851 TI - Physician or clinical inertia: what is it? Is it really a problem? And what can be done about it? PMID- 19125852 TI - All thiazide-like diuretics are not chlorthalidone: putting the ACCOMPLISH study into perspective. PMID- 19125853 TI - Altered molecular weight forms of adiponectin in hypertension. AB - An important link between adiponectin and hypertension has been proposed in clinical studies. In the circulation, adiponectin is predominantly present in multimeric complexes, of which high-molecular weight (HMW) adiponectin is thought to represent the biological active form. The authors investigated which role the different multimeric adiponectin isoforms play in context with hypertension as compared to total adiponectin levels. Fifty (19 normotensive/31 hypertensive) patients were included in the study. Total adiponectin and adiponectin multimers were determined by enzyme-linked immunosorbent assay and western blot. The authors analyzed associations between adiponectin multimer levels and blood pressure. Total adiponectin concentrations were not significantly different between hypertensive and normotensive patients (6.8+/-2.3 vs 7.5+/-4.2 microg/mL). HMW adiponectin was significantly lower (P<.05) and low-molecular weight adiponectin was significantly higher (P<.01) in hypertensive than in normotensive persons (3.8+/-1.7 vs 5.2+/-3.0 microg/mL and 0.9+/-0.5 vs 1.8+/ 0.9, respectively). Low molecular weight was an independent predictor for the presence of hypertension (effect coefficient: 0.160-0.445; P<.001) in multivariate analyses. These results suggest that the composition of the molecular weight forms of adiponectin in hypertension are characterized by reduced HMW adiponectin, the proposed major active form of adiponectin, and increased low-molecular weight adiponectin. Moreover, the latter represents an independent predictor of prevalent hypertension, suggesting an association between adiponectin multimer composition and hypertension. PMID- 19125854 TI - Pulse wave velocity as a marker of severity of coronary artery disease. AB - To determine whether pulse wave velocity (PWV) as a measure of arterial stiffness is a marker of coronary artery diseases (CAD), the authors did a cross-sectional study in 92 patients undergoing coronary angiography for suspected CAD. Arterial stiffness was assessed through recording PWV from the left carotid-right femoral arteries using an automated machine. The mean PWV was higher in patients with CAD than in those without CAD (11.13+/-0.91 vs 8.14+/-1.25 m/sec; P<.001). When the severity of CAD was expressed as 1-, 2-, and multiple-vessel disease, there was a significant association between the severity of CAD and PWV. PWV differed significantly with different categorical severity of CAD even when age and total cholesterol were controlled for. In a univariable analysis, PWV was higher with higher systolic blood pressure (P<.004). The authors conclude that arterial stiffness measured through PWV is an independent and complementary cardiovascular risk marker. PMID- 19125855 TI - The use of a single-pill calcium channel blocker/statin combination in the management of hypertension and dyslipidemia: a randomized, placebo-controlled, multicenter study. AB - Poor control of hypertension or dyslipidemia may at least in part be due to these risk factors being treated in isolation. The Caduet in Untreated Subjects Population (CUSP) trial was an 8-week, randomized, double-blind, placebo controlled trial evaluating the efficacy/safety of the combination of a calcium channel blocker (amlodipine besylate) and a statin (atorvastatin calcium) in a single-pill form (5/20 mg) plus therapeutic lifestyle changes (TLC) compared with placebo plus TLC in patients with comorbid hypertension and dyslipidemia without evidence of cardiovascular disease. At week 4, additional antihypertensive/lipid lowering medication was permitted. The primary end point was the proportion of patients in whom the dual goal of blood pressure (<140/90 mm Hg) and low-density lipoprotein cholesterol control (<100 mg/dL) was met at week 4. This dual goal attainment was significantly greater with amlodipine/atorvastatin plus TLC compared with placebo plus TLC at week 4 (47.6% vs 1.7%; P<.001), with further improvements at week 8. Most adverse events were mild to moderate. Therapy with single-pill amlodipine/atorvastatin plus TLC in these patients significantly increased dual blood pressure/low-density lipoprotein cholesterol goal attainment compared with placebo plus TLC. PMID- 19125856 TI - Lifestyle changes and blood pressure control: a community-based cross-sectional survey (2006 Ontario Survey on the Prevalence and Control of Hypertension). AB - To evaluate lifestyle changes and their impact on hypertension control in a sample of hypertensive respondents in Ontario, Canada, diet, physical activity, and other nonpharmacologic measures were recorded using a structured questionnaire during the 2006 Ontario Survey on the Prevalence and Control of Hypertension. Responses were weighted to the total adult population of 7,996,653 in Ontario. The prevalence of hypertension was 21%; 42% of hypertensive persons received therapy with antihypertensive drugs and lifestyle changes, and 41% received therapy with drugs only. Blood pressure was controlled in 85% of respondents who used only drugs and in 78% of those who stated that they received therapy with combined drug treatment and lifestyle changes. Fewer than half of hypertensive respondents practiced lifestyle changes (in combination with drug treatment) for blood pressure control. Lifestyle measures in addition to medication use did not result in better control of hypertension compared to only medication use. PMID- 19125857 TI - Progress and controversies: treating obesity and insulin resistance in the context of hypertension. AB - Improvements in hypertension treatment and control are challenged by the increasing incidence of metabolic risk factors for hypertension, in particular, obesity and insulin resistance. Such risk factors can increase the severity of hypertension and can interact via a multitude of hormonal and inflammatory pathways. Their presence may affect antihypertensive agent choice with regard to antihypertensive efficacy as well as potential synergistic or antagonistic effects on inflammatory status and progression to diabetes. Furthermore, an increasing number of pharmacologic options are available to promote weight loss and insulin sensitivity that may affect blood pressure directly and indirectly. This review considers the metabolic basis for the complex interactions of hypertension with obesity and insulin resistance, and it assesses the clinical evidence for an impact of weight loss and insulin-sensitizing treatment on blood pressure. Awareness of these pathophysiologic interrelations and their implications for treatment are likely to be of increasing importance for successful blood pressure management. PMID- 19125858 TI - The expectant model of hypertension management: setting expectations to achieve blood pressure control--applying a business principle to hypertension management. PMID- 19125859 TI - Analysis of recent papers in hypertension. Lack of legacy effect with more intensive blood pressure control in the long-term follow-up of the United Kingdom Prospective Diabetes Study. PMID- 19125862 TI - Is enough being done to control hypertension? An evaluation of blood pressure control patterns in a community-based setting. PMID- 19125864 TI - Gray matter deficits in bipolar disorder are associated with genetic variability at interleukin-1 beta gene (2q13). AB - Twin, family and recent molecular studies support the hypothesis of genetic overlapping between schizophrenia and bipolar disorder. Brain structural features shared by both psychiatric disorders might be the phenotypic expression of a common genetic risk background. Interleukin-1 (IL-1) cluster (chromosome 2q13) genetic variability, previously associated with an increased risk both for schizophrenia and for bipolar disorder, has been also associated with gray matter (GM) deficits, ventricular enlargement and hypoactivity of prefrontal cortex in schizophrenia. The aim of the present study was to analyze the influence of IL-1 cluster on brain morphology in bipolar disorder. Genetic variability at IL-1B and IL-1RN genes was analyzed in 20 DSM-IV (Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition) bipolar patients. Magnetic resonance imaging (MRI) measurements were obtained for whole-brain GM and white matter, dorsolateral prefrontal cortex (DLPFC), superior temporal gyrus, hippocampus and lateral ventricles. MRI data were corrected for age and cranial size using regression parameters from a group of 45 healthy subjects. A -511C/T polymorphism (rs16944) of IL-1B gene was associated with whole-brain GM deficits (P = 0.031) and left DLPFCGM deficits (P = 0.047) in bipolar disorder patients. These findings support the hypothesis of IL-1 cluster variability as a shared genetic risk factor contributing to GM deficits both in bipolar disorder and in schizophrenia. Independent replication in larger samples would be of interest to confirm these results. PMID- 19125863 TI - Methyl-CpG-binding protein 2 polymorphisms and vulnerability to autism. AB - The methyl-binding protein gene, MECP2, is a candidate for involvement in autism through its implication as a major causative factor in Rett syndrome that has similarities to autism. Rare mutations in MECP2 have also been identified in autistic individuals. We have examined the possible broader involvement of MECP2 as a predisposing factor in the disorder. Analysis of polymorphic markers spanning the gene and comprising both microsatellites and single nucleotide polymorphisms (SNPs) by the transmission disequilibrium test in two collections of families (219 in total), one in the USA and one in the UK, has provided evidence for significant association (P = 0.009) for a three-marker SNP haplotype of MECP2 with autism/autism spectrum disorders. This association is supported by association of both Single Sequence Repeat (SSR) and SNP single markers located at the 3' end of the MECP2 locus and flanking sequence, the most significant being that of an indel marker located in intron 2 (P = 0.001 - Bonferroni corrected P = 0.006). This suggests that one or more functional variants of MECP2 existing at significant frequencies in the population may confer increased risk of autism/autism spectrum disorders and warrants further investigation in additional independent samples. PMID- 19125865 TI - Partial dissociation of molecular and behavioral measures of song habituation in adult zebra finches. AB - Initial playback of recorded birdsong triggers a number of responses in zebra finches, including overt listening behavior and ERK pathway-dependent activation of zenk gene transcription in the auditory lobule of the forebrain. Repetition of one song stimulus leads to persistent habituation of these responses, as measured by subsequent presentations 1 day later. In this study, we examined the causal relationships between behavioral and molecular (ERK/zenk) habituation. In a within-subject comparison, we found a strong correlation with the level of prior training for both responses (duration of behavioral listening and magnitude of zenk expression), but little correlation between these responses for birds within the same treatment group. We then tested the hypothesis that ERK/zenk activation during training is necessary for the development of habituation measured 1 day later. Cannula-directed infusion of a pharmacological inhibitor of ERK activation (U0126) immediately before training blocked the development of habituation of the zenk gene response. However, measurement of the effect on behavioral habituation was confounded because birds that were infused with a non-active drug analogue (U0124) showed a decreased response 1 day later, even to novel songs. We conclude that the behavioral response to song stimulation is strongly influenced by factors other than song familiarity, whereas the zenk response in the forebrain may be a more accurate indicator of actual experience hearing a particular song. PMID- 19125866 TI - Molecular responses of the Ts65Dn and Ts1Cje mouse models of Down syndrome to MK 801. AB - Down syndrome (DS), caused by trisomy of human chromosome 21 (chr21), is the most common genetic cause of intellectual disability. The Ts65Dn mouse model of DS is trisomic for orthologs of 94 chr21-encoded, confirmed protein-coding genes and displays a number of behavioral deficits. Recently, Ts65Dn mice were shown to be hypersensitive to the locomotor stimulatory effects of the high-affinity N-methyl d-aspartate (NMDA) receptor (NMDAR) channel blocker, MK-801. This is consistent with the functions of several chr21 proteins that are predicted directly or indirectly to impact NMDAR function or NMDAR-mediated signaling. In this study, we show that a second mouse model of DS, the Ts1Cje, which is trisomic for 70 protein-coding genes, is also hypersensitive to MK-801. To investigate the molecular basis for the responses to MK-801, we have measured levels of a subset of chr21 and phosphorylated non-chr21 proteins, in the cortex and hippocampus of Ts65Dn and Ts1Cje mice and euploid controls, with and without treatment with MK 801. We show that in euploid mice, the chr21-encoded proteins, TIAM1 and DYRK1A, and phosphorylation of AKT, ERK1/2 and the transcription factor ELK are involved in the MK-801 response. However, in both Ts65Dn and Ts1Cje mice, levels of phosphorylation are constitutively elevated in naive, unstimulated mice, and the MK-801-induced changes in TIAM1 and DYRK1A and in phosphorylation are either absent or abnormal, with both genotype and brain-region-specific patterns. These results emphasize the complexities of the pathway perturbations that arise with segmental trisomy. PMID- 19125874 TI - Tonabersat inhibits trigeminal ganglion neuronal-satellite glial cell signaling. AB - BACKGROUND: Sensitization and activation of trigeminal neurons are implicated in the underlying pathology of migraine, acute sinusitis, and allergic rhinitis. Cell bodies of trigeminal neurons that provide sensory innervation of the dura and nasal mucosa reside in the trigeminal ganglion in association with satellite glial cells where they communicate via gap junctions. Gap junctions, channels formed by connexins, modulate the excitability state of both neurons and glia under pathological conditions. Tonabersat, a compound being tested as an antimigraine drug, is thought to block gap junction activity. OBJECTIVE: To investigate the cellular events within trigeminal ganglia that may account for the significant comorbidity of migraine and rhinosinusitis and determine the effect of tonabersat on neuron-satellite glia communication. METHODS: Sprague Dawley rats injected with True Blue were used to localize neuronal cell bodies in the ganglion and study neuron-glia signaling via gap junctions in the trigeminal ganglion. Dye coupling studies were conducted under basal conditions and in response to tumor necrosis factor-alpha injection into the whisker pad and/or capsaicin injection into the eyebrow. Changes in connexin 26 and active p38 levels were determined by immunohistochemistry. In addition, the effect of tonabersat prior to chemical stimulation on gap junction activity and expression of connexins and active p38 was investigated. RESULTS: Injection of tumor necrosis factor-alpha, a cytokine implicated in the pathology of acute sinusitis and allergic rhinitis, into the V2 region was shown to lower the amount of capsaicin required to stimulate neurons located in the V1 region of the ganglion. While injection of tumor necrosis factor-alpha into the whisker pad or capsaicin injection into the eyebrow alone did not cause increased dye movement, the combination of both stimuli greatly increased neuron-satellite glia communication via gap junctions in both V1 and V2 regions. The change in gap junction activity was accompanied by increased expression of connexin 26 and active p38 levels in both neurons and satellite glia in V1 and V2 regions. Pretreatment with tonabersat inhibited gap junction communication between neurons and satellite glia and blocked the increase in connexin 26 and active p38 levels in response to injection of both tumor necrosis factor-alpha (V2) and capsaicin (V1). CONCLUSIONS: We propose that increased levels of tumor necrosis factor-alpha, as reported during acute sinusitis and allergic rhinitis, reduces the amount of capsaicin necessary to stimulate V1 neurons that leads to cellular changes in both V1 and V2 regions. The cellular events observed in this study may help to explain, in part, the significant comorbidity reported with migraine and rhinosinusitis. In addition, we have provided evidence to suggest that tonabersat can prevent increased neuron-satellite glia signaling and, thus, may be useful in the treatment of migraine, acute sinusitis, and allergic rhinitis. PMID- 19125875 TI - Clinical significance of brush allodynia in emergency patients with migraine. AB - BACKGROUND: Cutaneous brush allodynia may be a practical and readily assessable marker of progression of an acute migraine attack. We determined the relative frequency of this finding in emergency department (ED) patients with acute migraine and tested the hypothesis that the presence of cutaneous brush allodynia prior to initial treatment in the ED could predict poor 2-hour and 24-hour pain intensity outcomes. METHODS: As part of a multicenter ED-based clinical trial testing the benefit of dexamethasone vs placebo for the adjuvant parenteral treatment of acute migraine, cutaneous brush allodynia was assessed prior to treatment using an established methodology. In addition to dexamethasone or placebo, all patients received intravenous metoclopramide + diphenhydramine as primary treatment for their migraine. Pain intensity outcomes were assessed in the ED 2 hours after medication administration and again by telephone 24 hours after medication administration. RESULTS: An assessment of cutaneous brush allodynia was performed in 182 migraineurs from 3 different EDs, of whom 26 (14%, 95% CI: 10-20%) had cutaneous brush allodynia. A pain-free state within 2 hours of medication administration was achieved by 46% of the allodynic patients and by 47% of the nonallodynic patients (P = .91). Median headache intensity over the 24 hours after ED discharge, as measured on a pain intensity scale from zero to 10, was 3 in the allodynic patients and 3 in the nonallodynic patients (P = .23). CONCLUSIONS: Cutaneous brush allodynia is an uncommon finding in the ED, occurring in fewer than 1 in 5 migraineurs. It does not seem to have prognostic relevance for the ED-based management of the acute migraine attack. PMID- 19125876 TI - Development and construct validation of the pharmacists' care of migraineurs scale. AB - OBJECTIVES: To develop the pharmacists' care of migraineurs scale (PCMS) and to evaluate its psychometric properties. BACKGROUND: Migraine is often managed suboptimally in primary care. Migraineurs frequently come into contact with community pharmacists, who have the opportunity to make a positive impact on migraineur treatment outcomes. A valid and reliable tool that measures and documents the care provided by pharmacists to migraineurs is critical to the development and evaluation of educational programs and interventions. METHODS: Relevant domains of pharmacist care and their respective composite items (behaviors) were identified through an extensive literature search and the use of 2 pharmacist and 2 migraineur focus groups sessions. The resultant 45 PCMS items composed a survey questionnaire mailed to a nationwide random sample of 6000 pharmacists. Data were subjected to an exploratory principal axis factoring procedure to discern the factor structure, and as such describe the latent domains composing the pharmacist caring behaviors constructs. RESULTS: A total of 580 usable responses were returned, with an additional 60 returned as undeliverable, thus yielding a response rate of 9.7%. Exploratory factor analysis using principal axis factoring yielded 9 factors. However, upon examining the scree plot, communalities, and factor loadings, a reanalysis forcing a 7-factor solution yielded a more interpretable and plausible factor structure. The 7 factor solution included the following domains: (1) empathy; (2) prospective drug utilization review for newly diagnosed migraineurs; (3) medication counseling; (4) nonpharmacologic treatment plan; (5) headache sufferer triage; (6) dissemination of public health information; (7) maintenance of knowledge on migraine. Following the application of scale purification procedures, the final instrument is composed of 41 items and demonstrated a Cronbach's alpha reliability of 0.947. Cronbach's alpha reliabilities for the 7 domains ranged from 0.67 to 0.91, indicative of good to excellent internal consistency reliabilities for all the domains. CONCLUSIONS: The PCMS demonstrated very good construct validity and reliability. While additional validity testing is warranted, the PCMS should allow for benchmarking in the evaluation of interventions designed to improve pharmacists' care to migraineurs and for identifying correlates to effective community pharmacist migraineur care. PMID- 19125877 TI - Prevalence and burden of headache and migraine in Germany. AB - OBJECTIVE: To determine prevalence and burden of headache and migraine in the general population in Germany including patterns of healthcare and medication use. METHODS: Telephone interviews were conducted on a representative sample of the general population in Germany aged >or=18 years (n = 7341). Migraine was diagnosed according to the International Headache Society criteria. RESULTS: One year prevalence was: headache 60.2%, migraine 10.6%, nonmigrainous severe headache 24.7% (women 66.6%, 15.6%, and 27.1%; men 53.0%, 5.3%, and 22.2%). Approximately 60% of headache sufferers reported severe headaches, 30% of which were migrainous. Migraineurs reported more often frequent headaches, disability, use of analgesics, and medical consultation than individuals with nonmigrainous severe headaches. Only 42% of migraineurs had consulted a physician and the majority relied exclusively on over-the-counter medication. CONCLUSION: Migraine accounts for a great part of the healthcare impact of headaches in Germany. However, the majority of migraineurs do not seek medical care and may not be optimally treated. PMID- 19125878 TI - Response of cluster headache to kudzu. AB - OBJECTIVE: To describe the self-treatment of cluster headache with kudzu. BACKGROUND: Many cluster headache patients take over-the-counter (OTC) kudzu extract in the belief that it helps their cluster attacks. Kudzu's actual efficacy has not been studied. METHODS: A database of cluster headache patients was questioned about their use of various alternative remedies to treat their cluster headache. Of 235 patients identified, 16 had used kudzu, consented to interviews, and provided medical records. RESULTS: In total, 11 (69%) experienced decreased intensity of attacks, 9 (56%) decreased frequency, and 5 (31%) decreased duration, with minimal side effects. CONCLUSION: Anecdotal evidence suggests that a component in OTC products labeled as kudzu may prove useful in managing cluster headache. This hypothesis should be tested with a randomized clinical trial. PMID- 19125879 TI - Inpatient treatment of status migraine with dihydroergotamine in children and adolescents. AB - OBJECTIVE: To assess the effectiveness of aggressive therapy of status migraine in children and adolescents. BACKGROUND: Inpatient management of pediatric status migraine and intractable headache is limited because of a lack of studies and guidelines. Adult treatment is often based on anecdotal experience, although a few controlled studies have been reported. Added to that is the discomfort of general pediatricians and neurologists in using available effective treatments in pediatric patients (such as dihydroergotamine: DHE). METHODS: Charts of all patients admitted to the neurology service, at Cincinnati Children's Hospital Medical Center-Department of Neurology, for inpatient treatment for intractable headache/status migraine over a 6-week period were reviewed. Demographics, evaluation, diagnosis, and treatment used were tabulated. Data on the effectiveness of the treatments provided were evaluated. Thirty-two total consecutive charts were retrospectively reviewed during that period. RESULTS: Upon discharge, 74.4% of the patients were headache-free. The mean severity of the pain upon discharge was 1.02 +/- 2.22 (using the 0-10 pain scale). CONCLUSION: From our review, DHE is very effective in treating and aborting an episode of status migraine and should be offered to children and adolescent patients who have failed their usual abortive therapy to prevent further severe disability that mainly affects their schooling and social activities. PMID- 19125880 TI - Verapamil for cluster headache. Clinical pharmacology and possible mode of action. AB - Verapamil is used mainly in cardiovascular diseases. High-dose verapamil (360-720 mg) is, however, currently the mainstay in the prophylactic treatment of cluster headache. The oral pharmacokinetics are variable. The pharmacodynamic effect of verapamil, the effect on blood pressure, also varies considerably among subjects. The dose of verapamil used for cluster headache is approximately double the dose used in cardiovascular disease, most likely because verapamil is a substrate for the efflux transporter P-glycoprotein in the blood-brain barrier. The access of verapamil to the central nervous system is therefore limited. The clinical use of verapamil in cluster headache is reviewed and several relevant drug interactions are mentioned. Finally, its possible mode of action in cluster headache is discussed. The effect of verapamil in cluster headache most likely takes place in the hypothalamus.Verapamil is an L-type calcium channel blocker but it is also a blocker of other calcium channels (T-, P-, and possibly N- and Q-type Ca(2+) channels) and the human ether-a-go-go-related gene potassium channel. With so many different actions of verapamil, it is impossible at the present time to single out a certain mode of action of the drug in cluster headache. PMID- 19125881 TI - Alcohol and cluster headaches. PMID- 19125882 TI - Exploring home allodynia testing for future clinical and scientific uses. AB - Allodynia, a painful response to a usually nonpainful stimulus, is common in chronic migraine. The evaluation of allodynia can be important clinically. Dynamic mechanical allodynia (brush) testing has been shown to be both a simple and reliable way to evaluate allodynia. In this study, we show that self administered brush allodynia testing at home is a feasible means of evaluating and recording allodynia in relationship to chronic migraine. PMID- 19125883 TI - Cluster headache during pregnancy: case report and literature review. AB - A 32-year-old pregnant woman presented with cluster headache (CH) during the third trimester of a normal pregnancy. Pure oxygen mask inhalation was ineffective, and intranasal lidocaine applications were realized associated with oral methylprednisolone, given at 1 mg per kg once daily. These treatments rendered the pain tolerable and the pregnancy went to its term with no consequence on the baby. This case of CH attack during pregnancy raises the issues of the influence of sexual hormonal changes in women with CH and the way to treat this disease in such circumstance. To date, there are no therapeutic guidelines available; this case suggests some possibilities. PMID- 19125885 TI - Migraine "chronification": what you can do. PMID- 19125886 TI - Tricks with tetramers: how to get the most from multimeric peptide-MHC. AB - The development of fluorochrome-conjugated peptide-major histocompatibility complex (pMHC) multimers in conjunction with continuing advances in flow cytometry has transformed the study of antigen-specific T cells by enabling their visualization, enumeration, phenotypic characterization and isolation from ex vivo samples. Here, we bring together and discuss some of the 'tricks' that can be used to get the most out of pMHC multimers. These include: (1) simple procedures that can substantially enhance the staining intensity of cognate T cells with pMHC multimers; (2) the use of pMHC multimers to stain T cells with very-low-affinity T-cell receptor (TCR)/pMHC interactions, such as those that typically predominate in tumour-specific responses; and (3) the physical grading and clonotypic dissection of antigen-specific T cells based on the affinity of their cognate TCR using mutant pMHC multimers in conjunction with new approaches to the molecular analysis of TCR gene expression. We also examine how soluble pMHC can be used to examine T-cell activation, manipulate T-cell responses and study allogeneic and superantigen interactions with TCRs. Finally, we discuss the problems that arise with pMHC class II (pMHCII) multimers because of the low affinity of TCR/pMHCII interactions and lack of 'coreceptor help'. PMID- 19125889 TI - Assessment, management, and prevention of early childhood caries. AB - PURPOSE: To discuss the role of primary care health providers in identifying infants and young children at risk for dental caries during well-child visits, in providing anticipatory guidance to parents and primary care givers of at-risk children, and in providing appropriate referrals for the timely establishment of a dental home. DATA SOURCES: The search included the following: Ovid MEDLINE, PubMed, American Academy of Pediatrics Web site, American Academy of Pediatric Dentistry Web site, and the American Dental Association Web site. The following search terms were used: dental caries prevention, caries process, caries balance, dental home, early childhood caries, oral health disparities, dental caries risk assessment, fluoride varnish, oral health anticipatory guidance. Search was limited to English language sources from 1990 through 2007. CONCLUSIONS: Dental caries is a preventable and reversible infectious disease process, yet it continues to be the single most common chronic disease of childhood. Despite a decrease in caries prevalence and a decrease in untreated tooth decay in 6-19 year-olds in the United States, a 15.2% increase in disease was noted among the nation's youngest children aged 2-5 years. Primary care health providers are uniquely positioned to play a significant role in the prevention of dental caries and are encouraged to complete certification courses in caries risk assessment, intervention, education, and referral. IMPLICATIONS FOR PRACTICE: Clinicians need to understand the dental caries process, including the process of enamel demineralization and remineralization, and the factors contributing to caries balance. The importance of early identification and intervention for infants and toddlers at high risk for dental caries and primary care health provider delivered anticipatory guidance during well-child care visits cannot be overestimated. PMID- 19125888 TI - Interleukin-17 in host defence against bacterial, mycobacterial and fungal pathogens. AB - The mammalian immune system is intricately regulated, allowing for potent pathogen-specific immunity to be rapidly activated in response to infection with a broad and diverse array of potential pathogens. As a result of their ability to differentiate into distinct effector lineages, CD4 T cells significantly contribute to pathogen-specific adaptive immune responses. Through the production of effector cytokines, CD4 T helper (Th) cells orchestrate the precise mobilization of specific immune cells to eradicate infection. The protective effects of the newly identified lineage of Th17 cells against pathogens like Klebsiella pneumoniae, Citrobacter rodentium and Candida albicans indicate the capacity of Th17 cells to confer protection against extracellular bacterial and fungal pathogens, filling a critical void in host immunity not covered by the classically described Th1 lineage that activates immunity to intracellular pathogens or the Th2 lineage that is important in protection against mucosal parasitic pathogens. Host defence by Th17 cells extends beyond protection against extracellular bacterial and fungal pathogens, as demonstrated in infections against intracellular bacteria like Listeria monocytogenes and Salmonella enterica, as well as Mycobacterium tuberculosis. Herein, we summarize both experimental data from mouse infection models and epidemiological studies in humans that demonstrate the protective effects of interleukin-17 and Th17 CD4 T cells in immunity to bacterial, mycobacterial and fungal pathogens. PMID- 19125890 TI - Reconceptualizing the core of nurse practitioner education and practice. AB - PURPOSE: The movement to the doctor of nursing practice (DNP) is progressing rapidly with new programs emerging and curricular documents being developed. We argue that the implementation of the DNP is a good move for nursing, provided that we use the opportunity to reconceptualize the core of advanced practice nursing, especially nurse practitioner (NP) practice. DATA SOURCES: Theory and research articles from nursing focused on advanced practice nursing, NPs, and doctoral education. CONCLUSIONS: The foundation of NP education is currently based essentially on borrowed or shared content in assessment, pharmacology, and pathophysiology. We argue that the heart and soul of nursing is in health promotion, both in healthy persons and in those dealing with chronic illness. Current master's programs do not prepare NPs to assume high-level practice focused on health promotion and disease management using the latest theoretical developments in health behavior change, behavioral sciences, exercise physiology, nutrition, and medical anthropology. Although these are touched upon in most NP programs, they do not represent the core science of NP education and need to be a critical part of any DNP program. IMPLICATIONS FOR PRACTICE: Ultimately, our vision is for NP care to be consistently "different," yet just as essential as physician care, leading to positive outcomes in health promotion and disease management. PMID- 19125887 TI - T-cell receptor binding affinities and kinetics: impact on T-cell activity and specificity. AB - The interaction between the T-cell receptor (TCR) and its peptide-major histocompatibility complex (pepMHC) ligand plays a critical role in determining the activity and specificity of the T cell. The binding properties associated with these interactions have now been studied in many systems, providing a framework for a mechanistic understanding of the initial events that govern T cell function. There have been various other reviews that have described the structural and biochemical features of TCR : pepMHC interactions. Here we provide an overview of four areas that directly impact our understanding of T-cell function, as viewed from the perspective of the TCR : pepMHC interaction: (1) relationships between T-cell activity and TCR : pepMHC binding parameters, (2) TCR affinity, avidity and clustering, (3) influence of coreceptors on pepMHC binding by TCRs and T-cell activity, and (4) impact of TCR binding affinity on antigenic peptide specificity. PMID- 19125891 TI - Perspectives of healthy elders on advance care planning. AB - PURPOSE: To present a qualitative study that explored the perspectives of healthy elders on advance care planning. DATA SOURCE: Data were gathered through four focus groups and a demographic questionnaire. The focus group data were analyzed using content analysis. A convenience sample of 20 healthy men and women, aged 60 94 years old, was drawn from one community senior center and two assisted living facilities. CONCLUSIONS: The data reveal five major themes: advance care planning is strongly influenced by concern for others; elders assume that preferences are known to their trusted friends, family, and providers, even in the absence of explicit communication with these people; elders value a healthcare system that supports provider time, focus, and continuity; being "known" to a provider is critical to comfort that advance care planning preferences will be respected; and elders are generally ready and eager to discuss advance care planning. Additional findings include: elders are better prepared for the event of death than the dying process; lawyers and financial planners play a prominent role in guiding elders through end-of-life decisions; and elders believe that the optimal time for advance care planning discussion is during periods of relative wellness. IMPLICATIONS FOR PRACTICE: Enhanced understanding of the patient perspective is key to incorporating advance care planning for healthy elders in the ideal milieu of primary care. PMID- 19125892 TI - Evaluation of a diabetes specialist-guided primary care diabetes treatment program. AB - PURPOSE: An initial pilot program demonstrated promising results in improvements in glycosylated hemoglobin (HbA(1c)), low-density lipoprotein cholesterol (LDL C), and systolic blood pressure (SBP) and prompted us to test these findings in a controlled trial. The purpose of the Diabetes-focused, Algorithm-directed care, Midlevel practitioner-administered, Electronically coached, Treatment (DAMET-2) program clinical trial was to investigate the benefits of a novel program for disseminating guidance in the treatment of diabetes from a central specialist clinic to primary care centers with access to midlevel provider services. DATA SOURCES: DAMET-2 included standardized treatment algorithms and education disseminated through computer-assisted and traditional methods associated with distance medicine. Two primary care practices were selected and subjects with diagnosed type 2 diabetes > or =6 months, > or =18 years of age with one or more cardiovascular risk factors (identified by chart review) were eligible for inclusion. Midlevel practitioners for subjects in the experimental group (N = 34) received training in American Diabetes Association treatment algorithms, had telephone consultations at 2- to 4-week intervals and bimonthly visits with diabetes specialists, and received treatment guidance within 24 h from remote diabetes specialists. Weekly diabetes clinics were made available to subjects in the experimental group. After 12 months, the last available subject data were extracted from the subjects' charts and compared to 12-month chart data from a control group (N = 101) that did not receive additional study services. CONCLUSIONS: Mean HbA(1c) values decreased from baseline by 0.46% in the active treatment group versus 0.06% in the control group; however, reductions in HbA(1c) did not achieve statistical significance potentially because of the small sample size of the experimental group. Mean SBP values were significantly reduced in both groups; however, LDL-C was only significantly reduced in the control group, where more aggressive use of statins may have had an effect. IMPLICATIONS FOR PRACTICE: Despite the inconsistencies in risk factor reduction from the pilot program, the DAMET-2 program provided insights regarding the importance of electronic records and provider notifications, patient adherence, prioritization of provider resources by risk factor level among patients, and access to self management education. PMID- 19125893 TI - Personal digital assistant use by nurse practitioners: a descriptive study. AB - PURPOSE: We sought to describe the prevalence and patterns of use of personal digital assistants (PDAs) among active nurse practitioners (NPs). DATA SOURCES: A descriptive correlational survey was conducted among NPs in the United States (N = 126). Participants were randomly selected from members of the American Academy of Nurse Practitioners who had listed a practice site on their application. CONCLUSIONS: Sixty-four percent of participants used PDAs. A drug reference was reported to be the most useful and frequently installed application. A large majority of PDA users believed that PDA use supported clinical decision making (91%), promoted patient safety (89%), and increased productivity (75%). Sixty-two percent predicted that PDA use would change their practice within the next 5 years. IMPLICATIONS FOR PRACTICE: As innovative PDA applications with potential to improve patient outcomes become increasingly available, handheld computer skills will be a fundamental practice competency. To prevent errors in clinical decision making with quick access to PDA reference materials, NPs must critically evaluate the legitimacy and worth of PDA software programs. There is a critical need to evaluate the effectiveness of PDA use in clinical settings and develop an evidence base to guide use of the PDA in solving clinical problems. PMID- 19125894 TI - Healthcare experiences of the homeless. AB - PURPOSE: To explore the healthcare experiences of homeless individuals and inform providers of the barriers created by the situation of homelessness. DATA SOURCES: This was a qualitative research study using a grounded theory approach. The sample included homeless individuals older than 18 years living in northeastern Wisconsin. CONCLUSIONS: This research provided rich insight into the healthcare experiences of the homeless. Five key conclusions were made: (a) the great majority of homeless people have an external locus of control; (b) most homeless individuals lack the necessary resources to meet their physical needs of shelter, air, water, and food; (c) most homeless individuals lack the financial resources to seek adequate health care; (d) access to resources is limited because of poor transportation, telephones, and mail; and (e) all those interviewed felt that healthcare providers lack compassion for the homeless. IMPLICATIONS FOR PRACTICE: Healthcare providers can use the concepts discovered in this study to help improve their skills and comfort level when working with homeless individuals. A decrease in acute illnesses and an increase in the effective management of chronic disease resulting in fewer long-term complications and medical costs because of these unnecessary complications could be seen. Healthcare professionals may also volunteer to become more involved with the care of the homeless if they are confident in their skills. Improving the health of the homeless in the community will result in improvements in the overall health of the community. PMID- 19125895 TI - Preventing coronary heart disease and stroke with aggressive statin therapy in older adults using a team management model. AB - PURPOSES: The purposes of this review were to highlight recent clinical trial results on the safety and efficacy of statin therapy in adults aged 65 years and older and to examine how nurse practitioners (NPs) and physician assistants (PAs) working together with physicians as a team can better serve the needs of this fastest growing subset of the U.S. population who are at high risk for cardiovascular events including stroke. DATA SOURCES: This report was compiled by reviewing the scientific literature on the safety and efficacy of statin therapy, including effects on cardiovascular, coronary, and stroke endpoints, with particular reference to clinical trial results in older patients. CONCLUSIONS: Recent guidelines emphasize that age alone is not a reason to withhold treatment with statin therapy in older adults. However, despite the evidence highlighted in this review demonstrating that older patients can safely achieve substantial benefit from statin therapy, studies have consistently shown an inverse relationship between age and treatment propensity. Often, older patients have multiple risk factors and many chronic comorbid conditions that seem to complicate and hinder treatment in many busy clinic practices. More extensive guidelines for the use of statin therapy in older patients, incorporating recommendations for treatment targets and developing a framework for the use of statins in stroke prevention, would assist care providers in the management of this high-risk population. Finally, NPs and PAs are both well educated and well placed to manage patients with chronic stable cardiovascular disease (CVD). IMPLICATIONS FOR PRACTICE: The implementation of a multidisciplinary team approach to identify at-risk individuals and provide them with education, counseling, and effective statin treatment will aid efforts to reduce the increased risk for morbidity and mortality associated with CVD in older adults. PMID- 19125896 TI - Management of asthma: new approaches to establishing control. AB - PURPOSE: The high burden of asthma indicates suboptimal control of this chronic condition. This review describes approaches for establishing asthma control based on an understanding of potential issues in the achievement and maintenance of asthma control, recent changes in asthma management guidelines that facilitate attainment of treatment goals, and the importance of the healthcare provider patient partnership to emphasize treatment based on asthma control. DATA SOURCES: Review of the published literature, asthma management guidelines, and patient asthma education resources. CONCLUSIONS: Asthma control is best achieved by patient-oriented versus disease-oriented management strategies that incorporate a combination of pharmacologic and nonpharmacologic treatment modalities. Tools that assess and monitor asthma may facilitate the achievement and maintenance of asthma control. Key components of an optimal management strategy include solid partnerships between healthcare providers and patients, comprehensive patient and caregiver education, personalized written asthma action plans, patient-reported evaluation of symptom control, appropriate drug therapy, strategies for improving compliance with asthma medication regimens, and a treatment algorithm that outlines the facets of asthma management. IMPLICATIONS FOR PRACTICE: Information presented in this article will guide nurse practitioners in helping patients with asthma achieve and maintain long-term disease control. PMID- 19125897 TI - Inhibiting the renin-angiotensin system: why and in which patients. AB - PURPOSE: To review the clinical benefits of inhibiting the renin-angiotensin system (RAS) through blood pressure (BP)-lowering and BP-independent mechanisms and to identify the benefits and potential limitations of RAS-blocking agents in various patient populations. DATA SOURCES: PubMed search using the key terms renin-angiotensin system, angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, aliskiren, heart failure, diabetes, and nephropathy. Current published guidelines from the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure, American Diabetes Association, and National Kidney Foundation were reviewed. CONCLUSION: Antihypertensive treatment with an agent that inhibits the RAS effectively lowers BP in a broad range of patients. Whether these agents improve clinical outcomes is the subject of ongoing investigation. Results of recent trials suggest that for patients with or at risk of high-risk conditions, such as heart failure or diabetes, risk reduction with RAS-blocking agents may be independent of BP reduction. Inhibition of the RAS may also reduce risk of renal impairment. IMPLICATIONS FOR PRACTICE: RAS-blocking agents are important in a variety of patient populations at high cardiovascular risk, but while angiotensin-converting enzyme inhibitors have proven benefits in some cases, angiotensin receptor blockers may be preferred in others. Direct renin inhibitors are currently being evaluated. The nurse practitioner should become familiar with the evidence for use of these agents to reduce risk and improve outcomes in specific populations. PMID- 19125898 TI - Inhaled corticosteroids for asthma: are they all the same? AB - OBJECTIVES: To assess similarities and differences among currently available inhaled corticosteroids (ICS) for treatment of asthma, with special emphasis on factors that may affect the relative safety of these medications. METHODS: PubMed was searched for relevant reviews and original articles. Information from these studies was synthesized and critically assessed. RESULTS: Differences in corticosteroid formulations and delivery systems can create variations in therapeutic efficacy. Chemical properties of the various corticosteroids may also affect their relative safety. Ciclesonide and beclomethasone dipropionate are administered as prodrugs activated by enzymes present in the lungs but not the oropharynx. Corticosteroid-specific adverse effects in the oropharynx are thus avoided, although formulation-specific effects may remain. Other adverse effects require systemic availability, either via the gastrointestinal tract or the lung. Once they enter the systemic circulation, all ICS are rapidly metabolized by the liver. Oral bioavailability of ICS such as fluticasone, ciclesonide and mometasone is minimal, as a result of their essentially complete first-pass metabolism in the liver. Ciclesonide also undergoes extrahepatic metabolism that eliminates it even more rapidly. Additionally, ciclesonide and mometasone exhibit very high levels of binding to serum proteins that reduces their ability to stimulate glucocorticoid receptors outside the lung. CONCLUSIONS: Despite acting by similar mechanisms, currently available ICS and their delivery systems differ in ways that can potentially affect both safety and therapeutic effectiveness for individual patients. PMID- 19125899 TI - Nateglinide and glibenclamide metabolic effects in naive type 2 diabetic patients treated with metformin. AB - BACKGROUND AND OBJECTIVE: Most antidiabetic agents target only one of several underlying causes of diabetes. The complementary actions of the glinides and the biguanides may give optimal glycemic control in patients with type 2 diabetes mellitus. The aim of the present study was to compare the effects of nateglinide plus metformin with glibenclamide plus metformin on glucose and lipid metabolism, and haemodynamic parameters in patients with type 2 diabetes mellitus. METHODS: We enrolled 248 type 2 diabetic patients. Patients were randomly assigned to receive nateglinide (n = 124) or glibenclamide (n = 124), after 6 months of run in, in which we titrated nateglinide (starting dose 180 mg/day), glibenclamide (starting dose 7.5 mg/day), and metformin (starting dose 1500 mg/day). The final doses were (mean +/- standard deviation), 300 +/- 60, 12.5 +/- 2.5, and 2500 +/- 500 mg/day, respectively. We followed these patients for 1 year after titration. We assessed body mass index (BMI), fasting (FPG) and post-prandial (PPG) plasma glucose, glycosylated haemoglobin (HbA(1c)), fasting (FPI) and post-prandial (PPI) plasma insulin, homeostasis model assessment (HOMA) index, and lipid profile [total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), triglycerides (Tg), apolipoprotein A-I (Apo A-I), and apolipoprotein B (Apo B)], systolic blood pressure (SBP), and diastolic blood pressure (DBP). All variables were evaluated at baseline and after 3 and 6 months in the run-in period, and at baseline, and after 3, 6, 9 and 12 months for both treatment groups. RESULTS AND DISCUSSION: Body mass index did not show any significant change during the study. We observed a significant improvement from baseline to 1 year on HbA(1c) (P < 0.01 vs. baseline and vs. glibenclamide group, respectively), FPG (P < 0.01 vs. baseline), PPG (P < 0.01 vs. baseline), and on HOMA index (P < 0.05 vs. baseline) in the nateglinide group. In the glibenclamide group, we found significant changes in HbA(1c) (P < 0.05 vs. baseline), FPG (P < 0.01 vs. baseline), PPG (P < 0.05 vs. baseline), and HOMA index (P < 0.05 vs. baseline). No significant change was observed in TC, LDL C, HDL-C, Tg, Apo A-I, Apo B, SBP, DBP and HR in either group after 3, 6, 9 and 12 months. These effects of nateglinide and glibenclamide on insulin-resistance parameters are in agreement with previous reports. Contrarily to previous reports, we did not observe any significant BP change in patients treated with glibenclamide. Although both nateglinide and glibenclamide attenuated PPG and HOMA index, they did not have significant effects on lipid metabolism, as already shown in subjects with type 2 diabetes and good glycemic control. CONCLUSION: Nateglinide improved glycemic control better than glibenclamide in combination with metformin. PMID- 19125900 TI - Nefopam hydrochloride compatibility and stability with selected proton pump inhibitors in bionolyte G5 injection for intravenous infusion. AB - BACKGROUND: The use of extemporaneously prepared admixtures of drugs must be supported by documentation of their chemical stability. OBJECTIVE: To assess the physical compatibility and the chemical stability of nefopam hydrochloride, a centrally acting non-opioid analgesic, when admixed with selected proton pump inhibitors (omeprazole, esomeprazole or pantoprazole), in bionolyte G5 injection for intravenous infusion. METHOD: Admixtures were assessed for periods of up to 72 h after storage at ambient temperature without protection from light and at +4 degrees C protected from light. A preparation was considered stable if the compounds of the mixture retained at least 90% of their original potency during the storage. Triplicate samples of nefopam and the selected proton pump inhibitors as well as the following mixtures (nefopam/omeprazole, nefopam/esomeprazole and nefopam/pantoprazole) were prepared in the concentrations required, in polypropylene bottles of bionolyte G5 injection. The physical compatibility was assessed by visual observation at each sampling interval. The chemical stability of the drugs was evaluated by high-performance liquid chromatography and by measurement of pH values. RESULTS: During refrigerated storage, nefopam as well as the selected proton pump inhibitors, when prepared separately in bionolyte G5 injection maintained chemical stability for up to 7 days. At ambient storage conditions, the protons pump inhibitors maintained chemical stability for 24 h, but thereafter their concentrations decreased significantly at day 1. Nefopam maintained chemical stability for up to 72 h at +25 degrees C. Nefopam/omeprazole and nefopam/esomeprazole mixtures in bionolyte were physically incompatible with the mixtures exhibiting a black colour. They underwent rapid and extensive loss, making the combination unacceptable within minutes of mixing. However, the nefopam/pantoprazole mixture was compatible over the study period, but with a reduced duration of the stability. CONCLUSION: Within the limits defined above, nefopam and the selected proton pump inhibitors may be prepared separately in advance in bionolyte G5 injection. The nefopam/pantoprazole mixture was stable for a short period, while the nefopam/omeprazole and the nefopam/esomeprazole mixtures were incompatible and unusable, immediately upon admixture. PMID- 19125901 TI - A prospective study evaluating clinical outcomes and costs of three NNRTI-based HAART regimens in Kerala, India. AB - OBJECTIVE: This prospective, observational, study evaluates the clinical outcomes, drug utilization patterns, and adherence to treatment of patients on highly active anti retroviral therapy (HAART) at a government institution in Kerala, India. METHODS: Patients who met criteria for treatment of HIV/AIDS were enrolled into the study, given free NNRTI-based combination therapy, and were followed for a period of 6 months. Data regarding demographics, clinical outcome, laboratory results, drug utilization, adherence and adverse effects were collected. Analysis was conducted utilizing descriptive statistics, anova, Fisher exact, andt-test. RESULTS: One hundred and forty-two patients with HIV-1 were enrolled in the study into three treatment groups. The mean age was 37.88 years, 64% of the patients were male, and 92% were married. Group 1 was given zidovudine, lamivudine, and nevirapine [n = 52 (37%)], group 2 was given lamivudine, stavudine, and nevirapine [n = 51 (36%)], and group 3 was given lamivudine, stavudine, and efavirenz [n = 39 (27%)]. The increase in CD4 was 107.46 (SD: 106.25). Mean medication adherence for the 104 patients who completed the study, was 90.7%; for group 1: 92.06%, group 2: 93.37%1, and group 3: 85.71% (P > 0.05). Forty (38%) patients have at least one adverse event to HARRT, with headache being the most common side effect (11.5%). Mortality rate was 3.5% during the course of the study. CONCLUSION: Provision of free NNRTI-based combination therapy to patients in Kerala, India, resulted in greater than 90% adherence leading to better clinical outcomes in terms of increasing CD4 counts and low mortality, for patients consistently attending a treatment clinic. PMID- 19125902 TI - Sequential therapy vs. standard triple therapies for Helicobacter pylori infection: a meta-analysis. AB - BACKGROUND: As standard triple therapies of achieve unsatisfactory eradication of Helicobacter pylori, several alternative regimens have been proposed. OBJECTIVES: To systematically evaluate whether sequential treatment eradicates H. pylori infection better than standard triple therapies and compare the risk of adverse events with these two regimens. METHODS: We searched electronic databases up to February 2008 for studies evaluating the efficacy of the 10-day sequential therapy vs. standard triple regimens for eradication of H. pylori. The pooled risk ratios (RR) and 95% confidence intervals (95% CI) were calculated. RESULTS: We identified 11 randomized trials, including eight full-text manuscripts and three abstracts. Pooled analysis demonstrated clear superiority of the sequential therapy over 7-day triple regimen with an RR of 1.23 (95% CI 1.19-1.27), and over 10-day triple regimen with a RR of 1.16 (95% CI 1.10-1.23). Adverse event rates were similar. For sequential therapy vs. 7-day triple therapies, RR = 0.96, 95% CI 0.70-1.31. CONCLUSIONS: Sequential therapy was associated with a higher eradication rate of H. pylori compared with both 7-day triple regimen and 10-day triple regimen. PMID- 19125903 TI - Pre-admission therapy for childhood acute diarrhoea--a hospital-based study. AB - Acute gastroenteritis (AGE) is a common illness among infants and children contributing to significant mortality and morbidity. As such, appropriate treatment received prior to hospital admission is of utmost importance. This retrospective observational study aimed to determine preadmission management in paediatric patients prior to hospital admission. Two hundred and twenty-two case notes of paediatric AGE patients were reviewed over a 12-month period. One hundred and fifty-four patients received medications prior to admission with 143 (92.9%) patients received known classes of medications. Antipyretic agents were the most commonly prescribed (69.2%), followed by antibiotics (38.5%), anti emetics (35.7%), oral rehydration salts (29.4%) and antidiarrhoeals (28.0%). The mean duration of stay in hospital was slightly shorter in patients, who received prior medications than those who did not (2.22 vs. 2.32 days respectively). Seventy per cent of children admitted for AGE were treated suboptimally prior to hospital admission with oral rehydration salts being largely under-utilized, despite their proven efficacy and safety. Sex, race and age had no influence on the type of preadmission treatment. A greater effort should be made to educate the general public in the appropriate treatment of AGE. PMID- 19125904 TI - Antibiotic use in pulmonology wards of Chinese children's hospitals: 2002-2006. AB - PURPOSE: To investigate the pattern of antibiotic use in the pulmonology wards of four children's hospitals in China from 2002 to 2006. METHODS: The Anatomical Therapeutic Chemical Classification/Defined Daily Doses (ATC/DDD) methodology was used. Aggregate data on antibiotic use (ATC code-J01) were expressed in numbers of DDD/100 bed-days for inpatients. RESULTS: The overall consumption of antibiotic drugs in the pulmonology wards of the four children's hospitals were 83.7, 49.9, 53.6 and 79.3 DDD/100 bed-days, respectively. Injectables were among the most widely used antibiotics. In addition, there was a considerable variation in both the amount and the pattern of antibiotics used in the respective hospitals' pulmonology wards. CONCLUSIONS: Wide variations were found between the four hospitals studied in the amount and type of antibiotics prescribed although the diseases treated appeared the same. This variation may have been due to differences in disease severity, regional diversity in bacterial resistance or variations in empiric treatment regimens. Despite the introduction of Chinese Ministry of Health Antibiotic Guidelines in 2004, the pattern of antibiotic use in individual hospitals remained unchanged from 2002 to 2006. Antibiotic utilization studies need be taken to the next level; comparing drugs used to treat specific diseases with guideline recommendations. In addition, educational and other interventions to ensure better compliance with guidelines are urgently required. PMID- 19125905 TI - The pharmaceutical industry and the German National Socialist Regime: I.G. Farben and pharmacological research. AB - Before the National Socialist party came to power, the German pharmaceutical industry constituted an international reference as far as the development of new medicines was concerned, having been responsible for synthetic analgesics (phenacetin, phenazones, acetylsalicylic acid), arsphenamine, barbiturates and sulfonamides. The year 1925 saw the founding of I.G. Farben (Interessen Gemeinschaft Farbenindustrie AG), a conglomerate of companies that would monopolize the country's chemical production and come to own all its major pharmaceutical industries. During the World War II, I.G. Farben participated in numerous operations associated with the criminal activities of the Nazi executive, including the use of slave labour in plants built close to concentration camps, such as that at Auschwitz. With regard to medical and pharmacological research projects, I.G. Farben became involved in experimental programmes using patients from the Nazi regime's euthanasia programmes and healthy subjects recruited without their consent from concentration camps, on whom various pharmacological substances were tested, including sulfamide and arsenical derivatives and other preparations whose composition is not precisely known (B-1012, B-1034, 3382 or Rutenol, 3582 or Acridine), generally in relation to the treatment of infectious diseases, such as typhus, erysipelas, scarlet fever or paratyphoid diarrhoea. Furthermore, I.G. Farben played a decisive role in the German army's chemical warfare programme, contributing to the development of the first two neurotoxic substances, later known as 'nerve agents', tabun and sarin. Some of these activities came to light as a result of the one the famous Nuremberg Trials in 1947, which saw 24 executives and scientists from I.G. Farben brought to justice for, among other offences, the use of slave labour in the concentration camps and forced experimentation with drugs on prisoners. PMID- 19125906 TI - Childhood abuse and treatment response in patients with irritable bowel syndrome: a post-hoc analysis of a 12-week, randomized, double-blind, placebo-controlled trial of paroxetine controlled release. AB - OBJECTIVE: Although irritable bowel syndrome (IBS) is frequently comorbid with childhood trauma, information on the clinical implications of this comorbidity is limited. We investigated whether a history of abuse was associated with response to treatment in a double blind, randomized, placebo controlled trial of paroxetine controlled release (CR) in IBS. METHODS: Seventy-two IBS subjects were randomized to receive paroxetine CR (dose 12.5-50 mg/day) or placebo for 12 weeks. Subject selection was independent of abuse history. Sixty-one subjects completed the Sexual and Physical Abuse Questionnaire about their childhood abuse history. IBS symptoms were recorded using the Interactive Voice Response System (IVRS). Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Perceived Stress Scale (PSS) and Clinical Global Impression (CGI) were also measured. The primary outcome was treatment response defined as > or =25% reduction in composite pain scores (CPS) on the IVRS from randomization to end of treatment. RESULTS: The rate of abuse history was 50.8% (n = 31/61). Baseline demographic clinical characteristics (CPS, BDI, BAI, PSS, CGI scores) were not associated with abuse history. After 12 weeks of treatment, subjects with abuse history showed significantly higher CPS (t = 2.422, P = 0.018) than subjects without a history and less mean change of CPS (t = 3.506, P = 0.001). In a logistic regression analysis, history of abuse did not predict treatment response as measured by > or =25% reduction in CPS (OR = 0.481, CI = 0.164-1.406, P = 0.181), while the drug status (paroxetine CR) was significantly associated with treatment response as defined by a CGI improvement score of 1-2 (OR = 12.121, CI = 2.923 50.271, P = 0.001). Abuse history did not predict CGI-I (Fisher's exact, P = 0.500) improvements during the trial. CONCLUSIONS: History of abuse did not appear to have any significant clinical correlates at baseline and did not predict treatment response. Further studies are needed to confirm whether SSRIs are effective in IBS patients irrespective of their abuse history. PMID- 19125907 TI - A limited sampling strategy for estimating mycophenolic acid area under the curve in adult heart transplant patients treated with concomitant cyclosporine. AB - OBJECTIVE: Heart transplantation studies have shown a relationship between the mycophenolic acid area under the curve (AUC) 0-12 h (MPA AUC(0-12h)) values and risk of acute rejection episodes and fewer side-effects in patient receiving cyclosporine during the first year post-transplant. However, measurement of full AUC is costly and time consuming and in this case it is an impractical approach to drug monitoring. Therefore, the authors describe a limited sampling strategy to estimate the MPA AUC(0-12h) value in adult heart transplant recipients. METHODS: Ninety MPA pharmacokinetic (PK) profiles were studied. The samples were collected immediately before and 0.5, 1, 1.5, 2, 2.5, 3, 4, 6, 9, 12 h after the morning dose of mycophenolate mofetil (MMF) following an overnight fast. PK profiles were determined at 6-8 weeks, 6, 12 months and more than 1 year after transplantation. Using stepwise multiple linear regression analysis a sampling strategy from 60 of PK profiles was obtained and next the bias and precision of the model were evaluated in another 30 PK profiles. RESULTS: The three-point model using C(0.5h), C(1h), C(2h) was found to be superior to all other models tested (r(2) = 0.841). The regression equation for AUC estimation which gave the best fit to this model is: 9.69 + 0.63C(0.5) + 0.61C(1) + 2.20C(2). Using that model 63 of the 90 (70%) full AUC values were estimated within 15% of their actual value. For the best-fit model, the mean prediction error was 3.2%, with 95% confidence intervals for prediction error to range from -42.2% to 40.3%. All other models which use one, two or three time-points over the first 2 h are poorer predictors of the full AUC than the model above. CONCLUSION: The proposed three time-point equation to estimate AUC will be helpful in optimizing immunosuppressive therapy in heart transplantation. PMID- 19125908 TI - Population pharmacokinetics of caffeine in healthy male adults using mixed effects models. AB - OBJECTIVE: Caffeine has been shown to maintain or improve the performance of individuals, but its pharmacokinetic profile for Asians has not been well characterized. In this study, a population pharmacokinetic model for describing the pharmacokinetics of caffeine in Singapore males was developed. The data were also analysed using non-compartmental models. METHODS: Data gathered from 59 male volunteers, who each ingested a single caffeine capsule in two clinical trials (3 or 5 mg/kg), were analysed via non-linear mixed-effects modelling. The participants' covariates, including age, body weight, and regularity of caffeinated-beverage consumption or smoking, were analysed in a stepwise fashion to identify their potential influence on caffeine pharmacokinetics. The final pharmacostatistical model was then subjected to stochastic simulation to predict the plasma concentrations of caffeine after oral (204, 340 and 476 mg) dosing regimens (repeated dosing every 6, 8 or 12 h) over a hypothetical 3-day period. RESULTS: The data were best described by a one-compartmental model with first order absorption and first-order elimination. Smoking status was an influential covariate for clearance: clearance (mL/min) = 110*SMOKE + 114, where SMOKE was 0 and 1 for the non-smoker and the smoker respectively. Interoccasion variability was smaller compared to interindividual variability in clearance, volume and absorption rate (27% vs. 33%, 10% vs. 15% and 23% vs. 51% respectively). The extrapolated elimination half-lives of caffeine in the non-smokers and the smokers were 4.3 +/- 1.5 and 3.0 +/- 0.7 h respectively. Dosing simulations indicated that dosing regimens of 340 mg (repeated every 8 h) and 476 mg (repeated every 6 h) should achieve population-averaged caffeine concentrations within the reported beneficial range (4.5-9 microg/mL) in the non-smokers and the smokers respectively over 72 h. CONCLUSION: The population pharmacokinetic model satisfactorily described the disposition and variability of caffeine in the data. Mixed-effects modelling showed that the dose of caffeine depended on cigarette smoking status. PMID- 19125909 TI - Amphotericin B removal by plasma exchange. AB - This case report adds pharmacokinetic knowledge regarding amphotericin B. Amphotericin B is highly protein bound. Plasma exchange removes 50-75% of a substance in plasma within 1-2 h, corresponding to an elimination half-life of 30 40 min. Amphotericin B reduction ratio by plasma exchange was 40% in this patient who had both liver and renal failure. PMID- 19125910 TI - Use of Tris-hydroxymethyl aminomethane in severe lactic acidosis due to highly active antiretroviral therapy: a case report. AB - BACKGROUND: Lactic acidosis is a rare, yet life-threatening adverse drug effect of highly active antiretroviral therapy (HAART), specifically stavudine and lamivudine. These nucleoside analogue reverse transcriptase inhibitors (NRTIs) are commonly used to treat patients infected with the human immunodeficiency virus (HIV). CASE: We report the use of Tris-hydroxymethyl aminomethane (THAM) to treat severe lactic acidosis due to HAART in a 50-year-old African-American woman. NRTIs can cause hyperlactinaemia by interfering with mitochondrial oxidative phosphorylation function, which normally removes H(+) generated by the hydrolysis of adenosine triphosphate. This side-effect is associated with a high mortality in patients infected with HIV. One explanation for this high mortality is that lactic acidosis is typically refractory to treatment with commonly used buffering agents. CONCLUSION: THAM generates serum bicarbonate, and reduces the level of carbon dioxide in arterial blood. Both of these qualities appear to make THAM an ideal agent for treating lactic acidosis caused by HAART. PMID- 19125911 TI - Chronic myeloid leukaemia in two multiple sclerosis patients on interferon beta 1a. AB - Haematological problems are commonly associated with use of beta-interferon in multiple sclerosis (MS) patients. However these problems are usually benign and are reversible when the drug is stopped. We describe two patients, with the diagnosis of MS, who developed leucocytosis. Cytogenetic studies showed the presence of the Philadelphia chromosome in all the cells analysed, confirming the diagnosis of chronic myeloid leukaemia (CML). The first patient currently on imatimib mesylate and interferon beta-1a is asymptomatic. For the second patient, interferon beta-1a was withdrawn after suspicion of CML. PMID- 19125912 TI - Increased levels of CA 125 and CA 19.9 serum tumour markers following cyclic combined hormone replacement therapy. AB - We report a case of increase in serum tumour markers CA 125 and CA 19.9 induced by cyclic combined hormone replacement therapy (HRT). A 52-year-old Caucasian post-menopausal woman presented with a slight enlargement of the right ovary and uterine fibromyomatosis. She was taking HRT for 4 years in a cyclic combined regimen of 2 mg oestradiol with 1 mg cyproterone acetate. The serum tumour markers occasionally measured were in normal range except CA 19.9 (997 U/mL; normal values 0.0-37) and CA 125 (85 U/mL; normal values 0.0-35). However, on one occasion, the CA 19.9 and CA 125 were high and then showed persistently high values (1005 and 81.3 U/mL, respectively). Radiodiagnostic investigations excluded any malignancies and a hysteroscopy showed endometrial thickening. After discontinuation of HRT, CA 125 levels returned to normal after 1 month, whereas CA 19.9 took 6 months to do so. Four months after the beginning subsequent therapy with over-the-counter phyto-oestrogens a new serum test showed an increase in CA 19.9 but CA 125 remained within the normal range. Phyto-oestrogen therapy was then interrupted and 1 month later CA 19.9 returned to normal. In this case, cyclic HRT was the probable cause of CA 19.9 and CA 125 increase. Positive dechallenge and subsequent CA 19.9 increase after phyto-oestrogen intake seem to confirm the role of oestrogens as the cause of the endometrial thickening through hormonal imbalance. Increased CA 19.9 and CA 125 levels in benign gynaecological conditions may be a source of misdiagnosis of malignant disease. PMID- 19125913 TI - The commoditization of hydrogeology. PMID- 19125915 TI - "An analytical solution for steady flow into a tunnel" by Shizhong Lei, January February 1999 issue, v. 37, no. 1: 23-26. PMID- 19125917 TI - Modeling analysis of ground water recharge potential on alluvial fans using limited data. AB - A modeling approach is developed to evaluate the potential for artificial recharge on alluvial fans in the Salinas Valley, California, using limited data of soil texture, soil hydraulic properties, and interwell stratigraphy. Promising areas for surface recharge are identified and mapped on a broad-scale using soil surveys, geologic investigations, permeability tests, and seasonal ground water response to rainfall and runoff. Two-dimensional representations of the vadose zone at selected sites are then constructed from drillers'logs and soil material types are estimated. Next, hydraulic properties are assigned to each soil material type by comparing them to laboratory-tested cores of similar soils taken from one site. Finally, water flow through the vadose zone is modeled in two dimensions at seven sites using a transient, finite-difference, variably saturated flow model. Average infiltration rates range from 0.84 to 1.54 cm/hr and recharge efficiency, the percentage of infiltrated water that reaches the water table, varies from 51% to 79%. Infiltration rates and recharge efficiency are found to be relatively insensitive to recharge basin ponding depth due to the thickness of the vadose zones modeled (31 to 84 m). The impact of artificial recharge on the Salinas Valley ground water basin is investigated by simulating the regional ground water response to surface spreading and streamflow augmentation with a recently calibrated, finite-element, ground water-surface water model for the basin. It was determined that a combined approach of surface recharge and streamflow augmentation significantly reduces the state of ground water overdraft and, to a lesser extent, reduces the rate of sea water intrusion. PMID- 19125918 TI - On the use of semi-logarithmic plots for baseflow separation. AB - Recession flow has long been considered a composite of exponential terms, where each exponential term represents a different source of water discharged from the watershed. The changing slopes in the semi-logarithmic plot of the discharge have been considered indicative of the decreasing contribution of surface runoff and interflow to the discharge. The results of this analysis show that the changing slope in the recession plot can be the consequence of baseflow drainage. This can invalidate the semi-logarithmic baseflow separation technique when applied to some hydrologic settings. PMID- 19125919 TI - Spatial and temporal characteristics of saline springs: Sea of Galilee, Israel. AB - Spatial and temporal characteristics of the saline springs that emerge along the western shore of the Sea of Galilee (Lake Kinneret) are analyzed. Three groups of onshore springs (Tiberias, Fuliya, and Tabgha) and two groups of offshore springs (Barbutim and Maagan), contribute saline water to the lake with concentrations in the range of 300 to 18,000 mg Cl/L, depending on location and season. It is well accepted that water emerging from these springs is a mixture of two endmembers: deep-seated saline ground water and shallow, fresh circulating ground water. Temporal trends of discharge rates and of chloride (representing the deep saline aquifer) and nitrate (representing the shallow fresh water aquifer) concentrations within each group of springs are presented. Results show the proportions of the two water bodies while mixing are time dependent. Discharge and concentration peaks in Tabgha springs precede those in Fuliya and Tiberias springs by approximately two months. An analytical solution shows that in Tabgha, variations of these parameters are mainly controlled by recharge variations in the Galilee, and follow an exponential function. In Fuliya and Tiberias, variations of these parameters are mainly dependent on lake level, and follow a sine-cosine function. The different patterns are attributed to different hydraulic properties of the discharge area. PMID- 19125920 TI - Role of macropores in solute transport under ponded water condition produced by laboratory simulated intense storms. AB - Soil macropores are widely studied because they are known to serve as avenues for rapid contaminant transport. Currently, it is understood that a macroporous medium develops regions of mobile and immobile water during flow. Consequently, solutes in the mobile phase bypass a considerable portion of the medium, thus limiting their natural attenuation. It is, however, not adequately documented in the literature what role the macropores play during intense rain events that result in variable ponding depths. Macropore flow under ponded water conditions was studied on two undisturbed soil columns taken from the loam soils of northeastern Iowa, U.S. The columns were placed on a four-segment outflow collector system. A slug of water labeled with Br was applied to the columns followed by simulation of 58 mm of rain on column 1 and 53.6 mm on column 2, in one hour, 30 minute, and 15 minute events. During the highest intensity rain event (15 minutes), the total volume of water that exited through the bottom of segments 1,2, 3, and 4 were 21.6%, 34.5%, 28.5%, and 15.4% in column 1, and 1.3%, 15.9%, 72.2%, and 10.5% in column 2, respectively. As the intensity of rain was increased from one hour to the 15 minute event, the total bromide mass recovery increased from 45% (29.2 mg) to 57% (36.9 mg) of the total application (65 mg) for column 1, and from 51 % (33.2 mg) to 59% (38.2 mg) for column 2, respectively. Finally, an examination of dye-stained high flow regions in column 1 demonstrated that macro-porosities made approximately 20% of the total cross sectional area of the column. These results suggest that solute transport behavior in macroporous soils can be impacted by ponding depths, and intense rain events can severely restrict contaminant degradation in soils. PMID- 19125921 TI - Use of combined Shewhart-CUSUM control charts for ground water monitoring applications. AB - Statistical properties of combined Shewhart-CUSUM control charts are examined in terms of the site-wide false positive rate, false negative rate, and average run length (i.e., the average number of sampling events that it takes to detect a significant exceedance). The effects of multiple comparisons (i.e., multiple monitoring wells and constituents), verification resampling, background sample size, and background updating are incorporated into the analysis. The effects of (1) verification resampling plans including no resampling, pass one resample, pass one of two resamples, and pass two of two resamples; (2) background sample sizes (n = 8,12,16,20, and 24); and (3) multiple comparisons (k = 1 to 500 future comparisons) were examined. In addition, various alternative control limit factors (h = SCL= 4.5, 5.0,5.5, and 6.0) were considered, where the control limit is x + hs. Results of simulations revealed that it is possible to balance site wide false positive and false negative rates at low levels using combined Shewhart-CUSUM control charts. The results revealed that it is important to select the appropriate multiplier, verification resampling plan, number of background samples, and number of intervening samples prior to updating background. PMID- 19125922 TI - Analysis of the quality of parameter estimates from repeated pumping and slug tests in a fractured porous aquifer system in Wonju, Korea. AB - Pumping and slug tests are widely used, relatively simple methods for estimating hydraulic conductivity and storage coefficient. This study tested the reproducibility of pumping and slug tests in estimating aquifer parameters in fractured or fractured porous media. The continuum concept is applied to represent the hydraulic properties by the conventional conductivity and storage coefficient. Repeated tests show discrepancies wider than can be attributed to probable measurement errors or incomplete curve fitting. The sources of discrepancy include turbulent head loss in an observation well, pumping well location, data acquisition period (pumping or recovery), selection of analysis method, test initiation mechanism, applied initial displacement, and selection of data segment for analysis. When we employ a specific test practice or choose a specific analysis method or data segment for the analysis, the resulting parameter estimates show characteristic patterns of distribution with respect to the mean value. This study analyzes the patterns and discusses how to improve the reliability of the parameter estimation. PMID- 19125923 TI - Predicted retardations of concentration fronts using a mass-balance approach. AB - Regardless of the chemical controls for a pollutant on sediment sorption or release from an immiscible organic liquid, a simple mass balance can be used to predict the retardation of the ground water pollutant front moving downstream from a source of constant composition. The retardation is the ratio of the total molar difference of pollutant across the front to the molar difference in ground water. The mass-balance approach presented here uses a flushing factor for the entire front. It is implicit in the commonly used retardation factor for pollutants undergoing linear sorption, but does not appear to have been used to predict retardation of concentration fronts involving nonlinear sorption or partitioning in hydrochemical studies. Yet the computations require only limited experimental or field data or the use of an algorithm describing sorption or partitioning of the pollutant. Close agreement was obtained between retardations predicted using the mass-balance approach and those reported from numerical modeling and column experiments in the literature. PMID- 19125924 TI - Natural attenuation of BTEX compounds: model development and field-scale application. AB - Benzene, toluene, ethyl benzene and xylene (BTEX) dissolved into ground water and migrated from a light nonaqueous phase liquid (LNAPL) source in a sandy aquifer near a petroleum, oil, and lubricants (POL) facility at Hill Air Force Base (AFB), Utah. Field observations indicated that microbially mediated BTEX degradation using multiple terminal electron-accepting processes including aerobic respiration, denitrification, Fe(III) reduction, sulfate reduction, and methanogenesis has occurred in the aquifer. To study the transport and transformation of dissolved BTEX compounds under natural conditions, a reactive flow and transport model incorporating biochemical multispecies interactions and BTEX was developed. The BTEX, oxygen, nitrate, Fe(II), sulfate, and methane plumes calculated by the model agree reasonably well with field observations. The first-order biodegradation rate constants, estimated based on model calibration are 0.051, 0.031, 0.005, 0.004, and 0.002 day(-1) for aerobic respiration, denitrification, Fe(III), sulfate reduction, and methanogenesis, respectively. The results of a sensitivity analysis show that the saturated aquifer thickness, hydraulic conductivity, and reaction rate constants are the most critical parameters controlling the natural attenuation of BTEX at this site. The hydraulic conductivity and aquifer thickness were found to be the key factors affecting the restoration of oxygen, nitrate, and sulfate after their interaction with the BTEX plume. The multispecies reactive transport modeling effort, describing BTEX degradation mediated by multiple electron-accepting processes, represents one of the few attempts to date to quantify a complete sequence of natural attenuation processes with a detailed field data set. Because the case study is representative of many petroleum-product contaminated sites, the results and insights obtained from this study are of general interest and relevance to other fuel-hydrocarbon natural attenuation sites. PMID- 19125925 TI - Characterizing a faulted aquifer by field testing and numerical simulation. AB - Faulted aquifers constitute one of the most complex geological environments for analysis and interpretation of hydraulic test data because of the inherent ability of faults to act not only as highly transmissive zones but also as hydraulic barriers. Previous studies of the fractured carbonate aquifer at Carleton University, Ottawa, Canada, characterized the flow regime as predominantly linear, but with limited radial nature, and undertook to analyze constant discharge test data using both radial and linear flow models. When used as direct input to a numerical model, the hydraulic parameters, calculated directly from hydraulic test data, were inappropriate and resulted in a poorly calibrated model. While our interpretation of the faulted aquifer remains linear radial in nature, parameter estimation by numerical simulation highlighted the presence of hydraulic barriers associated with the faults. These barriers are not readily apparent in the constant discharge test data and act to modify the hydraulic test curves at early to mid time, leading to incorrect estimates of the hydraulic parameters. This paper describes the conceptual model and the numerical approach, and demonstrates the importance of using transient simulations for model calibration. PMID- 19125926 TI - Application of flowmeter and depth-dependent water quality data for improved production well construction. AB - Ground water production wells commonly are designed to maximize well yield and, therefore, may be screened over several water-bearing zones. These water-bearing zones usually are identified, and their hydrogeologic characteristics and water quality are inferred, on the basis of indirect data such as geologic and geophysical logs. Production well designs based on these data may result in wells that are drilled deeper than necessary and are screened through zones having low permeability or poor-quality ground water. In this study, we examined the application of flowmeter logging and depth-dependent water quality samples for the improved design of production wells in a complex hydrogeologic setting. As a demonstration of these techniques, a flowmeter log and depth-dependent water quality data were collected from a long-screened production well within a multilayered coastal aquifer system in the Santa Clara-Calleguas Basin, Ventura County, California. Results showed that the well yields most of its water from four zones that constitute 58% of the screened interval. The importance of these zones to well yield was not readily discernible from indirect geologic or geophysical data. The flowmeter logs and downhole water quality data also show that small quantities of poor-quality water could degrade the overall quality of water from the well. The data obtained from one well can be applied to other proposed wells in the same hydrologic basin. The application of flowmeter and depth-dependent water quality data to well design can reduce installation costs and improve the quantity and quality of water produced from wells in complex multiple-aquifer systems. PMID- 19125927 TI - Sensitivity study on hydraulic well testing inversion using simulated annealing. AB - Cluster variable aperture (CVA) simulated annealing has been used as an inversion technique to construct fluid flow models of fractured formations based on transient pressure data from hydraulic tests. A two-dimensional fracture network system is represented as a filled regular lattice of fracture elements. The algorithm iteratively changes element apertures for a cluster of fracture elements in order to improve the match to observed pressure transients. Aperture size is chosen randomly from a list of discrete apertures. The cluster size is held constant throughout the iterations. Since hydraulic inversion is inherently nonunique, it is important to use additional information. We investigated the relationship between the scale of heterogeneity and the optimal cluster size and shape to enhance convergence of the inversion and improve the results. In a spatially correlated transmissivity field, a cluster size corresponding to about 20 % to 40 % of the practical range of the spatial correlation is optimal. Inversion results of the Raymond test site data are also presented and based on an optimal cluster size; the practical range of the spatial correlation is estimated to be 5 to 10 m. PMID- 19125928 TI - Simple probabilistic and statistical risk calculations in an aquifer. AB - An objective methodology is proposed for assessing potential ground water yield on the basis of the randomness inherent in hydrogeological parameters as a result of uncertainty. Randomness is used here in the statistical sense to describe the ground water phenomenon on a regional basis. A systematic approach to selecting aquifer parameters is presented that leads to reliable ground water storage and subsurface flow estimates by incorporating a certain risk level. In addition, the averages of the specific ground water capacity and subsurface flow rate are calculated through a perturbation approach. This study demonstrates that cross correlations between basic hydrogeologic variables might underestimate or overestimate model parameter values. Furthermore, the results of aquifer storage and subsurface flow estimates are obtained in the form of probability distribution functions. It is proved that the surface flow rates are presented by a log-normal distribution. Contrary to conventional point estimate, a simple and objective methodology is proposed in order to obtain the most probable interval estimates. Accordingly, projections of likely future ground water storage and flow rates can be readily calculated. PMID- 19125929 TI - Monitoring the hydraulic performance of a containment system with significant barometric pressure effects. AB - Under certain circumstances, wells in unconfined aquifers can display significant water level fluctuations in response to changes in barometric pressure. This is illustrated by Hare and Morse (1997) at a site where a portion of the unconfined aquifer is isolated by a soil-bentonite cutoff wall and clay cap. A relief well located within the containment system displays water level fluctuations that mirror barometric pressure changes. Water levels fluctuated 0.37 m in response to barometric pressure changes of 2.87 centimeters mercury (cm-Hg), representing a barometric efficiency of 93.6%. As described in this paper, the short-term variability in water level elevations inside the containment system had to be considered to develop a reliable post-enhancement performance monitoring program. The approach that was ultimately selected involves correcting the water level elevations obtained in the relief well within the containment system for the effects of barometric pressure changes prior to comparison with the water level elevations in an observation well in the aquifer outside the system. The reliability of the post-enhancement performance monitoring program is improved further by simply requiring that any decision to enhance the containment system be based on the water level measurements obtained during two consecutive months. Using this approach, the probability that the containment system's performance will erroneously be deemed unacceptable is low. The post-enhancement performance monitoring program also requires no extra field work and does not involve any specialized equipment, which helps to keep operation and maintenance costs to a minimum. PMID- 19125930 TI - Skin localization at wells drilled in a vertical fault zone. AB - The efficiency of wells drilled into fractured zones that act as draining conduits in hard rock aquifers can be strongly reduced by skin effects. In such aquifers, skin effects (well losses) can appear in close proximity to the well and/or at the contact between fracture and formation. In this paper it is shown how to locate those skin conditions in the case of two wells that have been drilled in a single vertical fault with infinite conductivity. PMID- 19125931 TI - Modeling ground water flow and radioactive transport in a fractured aquifer. AB - Three-dimensional numerical modeling is used to characterize ground water flow and contaminant transport at the Shoal nuclear test site in north-central Nevada. The fractured rock aquifer at the site is modeled using an equivalent porous medium approach. Field data are used to characterize the fracture system into classes: large, medium, and no/small fracture zones. Hydraulic conductivities are assigned based on discrete interval measurements. Contaminants from the Shoal test are assumed to all be located within the cavity. Several challenging issues are addressed in this study. Radionuclides are apportioned between surface deposits and volume deposits in nuclear melt glass, based on their volatility and previous observations. Surface-deposited radionuclides are released hydraulically after equilibration of the cavity with the surrounding ground water system, and as a function of ground water flow through the higher-porosity cavity into the low-porosity surrounding aquifer. Processes that are modeled include the release functions, retardation, radioactive decay, prompt injection, and ingrowth of daughter products. Prompt injection of radionuclides away from the cavity is found to increase the arrival of mass at the control plane but is not found to significantly impact calculated concentrations due to increased spreading. Behavior of the other radionuclides is affected by the slow chemical release and retardation behavior. The transport calculations are sensitive to many flow and transport parameters. Most important are the heterogeneity of the flow field and effective porosity. The effect of porosity in radioactive decay is crucial and has not been adequately addressed in the literature. For reactive solutes, retardation and the glass dissolution rate are also critical. PMID- 19125932 TI - A field study of nonequilibrium and facilitated transport of Cd in an alluvial gravel aquifer. AB - A natural-gradient tracer experiment was conducted to study Cd transport in an alluvial gravel aquifer. Both a conservative tracer and Cd exhibited tailing in their breakthrough curves (BTCs), indicating the presence of nonequilibrium transport. Solute transport was evaluated using a three-dimensional nonequilibrium analytical model, and the results were compared with those obtained from a previous laboratory study. At similar flow velocities, the field Cd data gave significantly lower retardation factors (R=7 to 30, median 22), higher fractions of instantaneous sorption sites (beta), and greater mass transfer coefficients (omega) than the laboratory data because of a high degree of aquifer heterogeneity, the presence of preferential flow, and the larger transport scale in the field conditions. Multiple peaks in the Cd BTCs were observed due to bacteria-facilitated and perhaps also colloid-facilitated transport. The early peaks showed narrower, more symmetric shapes with higher concentrations than the later peaks, and compared well to those of the bacterial BTCs. The multiple peaks of the Cd BTCs imply that a significant fraction of Cd could travel with little, if any, retardation over a 20 to 40 m travel distance when Cd and bacteria coexist in a contamination event. PMID- 19125933 TI - Haemophilia care: from paediatrics to geriatrics. Introductory note. PMID- 19125934 TI - Haemophilia care then, now and in the future. AB - Epidemiological data show the benefits of dramatically improved haemophilia care in all life-stages. There are improved administration techniques and dosing regimens, a shift from on-demand treatment to prophylaxis, successful treatment protocols for immune tolerance induction in patients with inhibitors and enhanced approaches to overall patient management. Improvements also include the introduction of virus inactivation methods for plasma derived clotting factor concentrates and the development of recombinant factor VIII therapy, which practically eliminated the risk of infectious disease transmission. Recombinant factor concentrates are recommended as treatment of choice by several guidelines today. All these developments have resulted in increased health-related quality of life and life expectancy in haemophilia patients, who are transitioning from childhood to adulthood with healthy joints and an overall healthy status today. Because of increased life expectancy, these patients are expected to experience age-related clinical problems that were not previously observed in this population. With respect to this, the spectrum of haemophilia care will be extended to diseases of older ages with the need of including further disciplines in comprehensive haemophilia care programmes. Despite these advances, the short half-life of factor VIII, requiring re-administration every 2 or 3 days and the development of inhibitors remains a challenge. Bayer's research and development currently focuses on the optimization of recombinant coagulation factors to address these challenges. Haemophilia care has experienced significant improvements within the past decades. Novel technologies and continued clinical research have facilitated the development of treatment regimen that resulted in dramatic increases in the life expectancy and quality of life of haemophilia patients. To set the scene for the following papers dealing with haemophilia care from paediatrics to geriatrics, developments behind these improvements and some aspects of future research will be presented in this paper. PMID- 19125935 TI - Haemophilia care in children--benefits of early prophylaxis for inhibitor prevention. AB - Haemophilia therapy is aimed at treating and preventing bleeding episodes and related complications and clinical studies have shown that regular prophylaxis, started at an early age, is able to reduce physical impairment from haemophilic arthropathy. Today, the development of anti-Factor VIII (FVIII) inhibitors is the most serious treatment-related complication of haemophilia therapy and a number of genetic and environmental risk factors have been identified in the past years. Clinical data show that early start of prophylaxis and the avoidance of intensive treatment periods may protect patients from inhibitor development. The mechanisms are not completely understood; yet, recent experimental data suggest that pro inflammatory or 'danger signals' may be involved in inducing tolerance vs. an effector immune response. So, exposure to a factor concentrate by itself may not be enough to trigger an immune response, while an intensive exposure to FVIII in the presence of such 'danger signals' can activate antigen-presenting cells, up regulating co-stimulatory signals for T lymphocytes and ultimately enhancing antibody production. The 'optimal' regimen for primary prophylaxis is still not identified and barriers to prophylaxis implementation remain relevant. Key issues include the optimal age at prophylaxis onset, the optimal dosage/schedule, the proper clinical and laboratory monitoring and patients' compliance. Practical approaches to early prophylaxis as implemented in the haemophilia centres in Milan and Bremen are discussed in this respect. PMID- 19125936 TI - Haemophilia care in adolescents--compliance and lifestyle issues. AB - Adolescence is characterized by simultaneous physical, psychological, social and sexual changes that compound the challenges faced by parents, health care providers and adolescent haemophilia patients themselves. Compliance with prophylactic factor replacement therapy frequently declines when patients pass from childhood to adolescence. Familiarity with long-term joint damage is lacking among the current generation of children who have grown up with prophylactic treatment and the tendency of teenagers to focus primarily on short-term goals increases the likelihood that regular prophylactic replacement therapy receives low priority. Most adolescents continue prophylactic treatment prior to physical or social activities because short-term goals are more likely to be perceived as relevant. The most important factor that influences compliance is support from parents, peers and caregivers, who provide encouragement and support active participation in health care management. During adolescence, personalized treatment strategies that suit the patient and his lifestyle are essential to ensure optimal outcomes. Physical activity is important for all adolescents and can contribute to better coordination, endurance, flexibility and strength. Physical training also contributes to healthier joints and reduces the risk of bleeding episodes in teenagers with haemophilia; however, the selection of an appropriate sport that minimizes the risk of injury and matches the patient's skill and needs is important. Children with haemophilia may have disease-related functional deficits and often exhibit subclinical findings in the joints; therefore an orthopaedic examination, fitness check and motion analysis may assist in guiding preventive physiotherapy and the choice of sport. PMID- 19125937 TI - Challenges and controversies in haemophilia care in adulthood. AB - Overall life expectancy and quality of life among persons with haemophilia have increased in recent years, primarily because of the advances in factor replacement therapy and better treatment of infectious diseases. Older haemophilic patients now face aging co-morbidities that are common in the general male population, such as cardiovascular or metabolic diseases, prostate hypertrophy and hepatic, prostate and other cancers. The prevalence of cardiovascular disease and incidence of vascular events among older haemophilic patients can be expected to increase and haemophilic patients may become prone to some cardiovascular risk factors, warranting preventative measures. The treatment of long-term complications of hepatitis C virus infection such as liver cirrhosis and hepatic cancer can be expected to be required in a large portion of the older haemophilia population for some years to come. Appropriate antiviral treatment and close monitoring for possible disease advancement will constitute an important part of routine medical care, and special considerations may be appropriate in conjunction with invasive procedures, chemo- or radiotherapy. At the moment, hard data on which to base the management of these conditions are largely lacking, but can be expected to increase dramatically in the coming decades. In the meantime, the ageing population of haemophilia patients should be offered the same comprehensive health care offered to the general population, which may require a restructuring of health care delivery. PMID- 19125939 TI - Preface. PMID- 19125938 TI - Spontaneous HBeAg seroconversion and loss of hepatitis B virus DNA after acute flare due to development of drug resistant mutants during entecavir monotherapy. AB - AIMS: Patients with chronic hepatitis B virus (HBV) infection under entecavir (ETV) treatment develop resistant mutants with viral rebound. Here, we report an interesting case of spontaneous loss of HBV-DNA and seroconversion following an acute flare after the development of ETV-resistant mutants. This patient received ETV after lamivudine breakthrough. METHODS: Cloning and sequence analysis of the HBV reverse transcriptase (RT) region were performed with seven samples during ETV therapy. In addition, two full-length HBV genomes derived from samples before and after the emergence of ETV resistance were sequenced. RESULTS: ETV resistant mutants appeared at week 228, with virological and biochemical rebound at the same time. Unexpectedly, HBeAg seroconversion occurred 8 weeks later. The viral load decreased and became undetectable from week 252. Analysis of HBV isolates in the patient at week 124 revealed that wild-type HBV was predominant at that time and ETV resistant mutants were not found among 20 clones. Interestingly, a new mutant type with rtL180M+rtT184L was found alongside rtL180M+rtT184L+rtM204V/I at week 228 and appeared to develop independently, according to the sequence analysis. In contrast to the previously identified ETV resistant mutants, it did not carry the rtM204V/I mutations. CONCLUSION: The data presented here indicates that the flare following the emergence of ETV resistant mutants may reflect immune-mediated control of HBV infection, leading to a spontaneous loss of HBV DNA and seroconversion. PMID- 19125940 TI - Long-term outcome of branched-chain amino acid treatment in patients with liver cirrhosis. AB - Clinical impact of protein-energy malnutrition (PEM) on the outcome of liver cirrhosis is well documented. As a candidate interventional modality to improve PEM in cirrhosis, effects of branched-chain amino acid (BCAA) supplementation on event-free survival and quality of life (QOL) was first reported by Yoshida et al. in 1989. Although critical arguments still continue regarding the effects of BCAA, several randomized trials in the last 5 years have brought positive results, and seem to have settled the discussion in a favorable direction for the efficacy of BCAA in liver cirrhosis. Actually, The European Society for Clinical Nutrition and Metabolism (ESPEN) upgraded the recommendation of BCAA supplementation in decompensated liver cirrhosis in the latest revision of its guidelines in 2006, by referring to the literatures from Italy and Japan. Particularly in these two long-term randomized studies with 1-2 years supplementation, event-free survival was estimated by employing composite endpoints such as aggravation of hepatic failure (ascites, peripheral edema, hepatic encephalopathy, and jaundice), rupture of esophageal or gastric varices, development of liver cancer, and death from any cause. Both trials agreed on the effect of BCAA to reduce the incidence of hepatic failure, thus contributing to the rise in the event-free survival. Quality of life is another essential marker of outcome survey. Marchesini, Muto, and Nakaya reported the improved QOL in cirrhotics with BCAA supplementation. In particular, quantitative analysis of QOL measured by Short Form 36 (SF-36) questionnaire demonstrated a significant recovery of general heath perception score in BCAA supplemented patients in a randomized trial. In this article, the long-term outcome of BCAA treatment in liver cirrhosis will be reviewed with its action mechanisms. In addition, the effects of BCAA treatment on the incidence of liver cancer in obese patients with type C liver cirrhosis, significance of obesity as a risk factor for type C liver cancer, and a possible role of Body Mass Index to estimate the histological grade of fat deposition in the liver will be briefly discussed. PMID- 19125941 TI - Role of branched-chain amino acids in management of cirrhosis and hepatocellular carcinoma. AB - Altered protein and energy metabolism is a hallmark of chronic liver disease, characterized by decreased plasma branched-chain amino acids (BCAA) and increased plasma aromatic amino acids (AAA). Overwhelming evidence has indicated that the incidence of complications of chronic liver disease increases with malnutrition. Hence nutritional management in patients with chronic liver disease must receive high priority. The use of BCAA supplementation has been a controversial subject. This review summarizes published results of BCAA supplementation as a nutritional therapy for patients with cirrhosis and hepatocellular carcinoma (HCC). On balance, it would be appropriate to conclude that BCAA are associated with decreased frequency of complications of cirrhosis and improved nutritional status when prescribed as a maintenance therapy for patients with cirrhosis. More studies are, however, required to identify those who might benefit most from BCAA supplementation. PMID- 19125942 TI - Pathophysiology of hepatic encephalopathy: The concept of synergism. AB - Hepatic encephalopathy (HE) remains a severe neuropsychiatric complication of liver failure. Neuropathological evaluation of material from patients who died in hepatic coma reveals morphologic changes primarily to astrocytes (cytotoxic edema, Alzheimer Type II astrocytosis) accompanied by discreet neuronal changes. Liver failure results in the accumulation in brain of neurotoxic compounds (ammonia, manganese, proinflammatory cytokines, mercaptans, octanoic acid) that may act synergistically to impair neuropsychiatric function. Ammonia and manganese act synergistically to activate mitochondrial benzodiazepine receptors leading to increased production of neuroactive steroids, many of which (allopregnanolone, THDOC) have potent neuroinhibitory properties resulting from activation of a neuromodulatory site on the GABA-A receptor ("increased GABAergic tone"). New evidence demonstrates that proinflammatory cytokines such as tumor necrosis factor alpha (TNFalpha) and the interleukins (IL-1beta, and IL-6) are produced not only by the liver but also by the brain in liver failure. Ammonia and proinflammatory cytokines generated either by intercurrent infection or from hepatocyte necrosis in liver failure act synergistically to decrease the capacity of astrocytes to remove glutamate from the brain extracellular space leading to the activation of glutamate (NMDA) receptors that in turn results in alterations of cell-cell signalling and hyperexcitability. Therapy for HE continues to rely heavily on strategies aimed at reduction of gut ammonia production, increased ammonia removal and, ultimately, liver transplantation. A more complete understanding of pathophysiologic mechanisms has led to novel potential strategies aimed at decreasing both GABAergic tone, glutamate (NMDA) receptor activation and proinflammatory cytokines. Such strategies hold promise for new therapies for HE in the near future. PMID- 19125943 TI - Diagnosis of sub-clinical hepatic encephalopathy by Neuropsychological Tests (NP tests). AB - AIM: At present, there are no generally accepted diagnostic criteria or methods for sub-clinical hepatic encephalopathy (SHE) associated with liver cirrhosis. We therefore developed an easily conducted computer-aided quantitative neuropsychological function test system for use in routine medical practice. METHODS: The system was used prepare basic values according to age in 542 healthy subjects, and the results were compared with 292 liver cirrhosis patients. The software is composed of eight tests: NCT-A, NCT-B, Figure Design Test, Digit Symbol Test, Block Design Test, and the Reaction Time-A, Reaction Time-B, and Reaction Time-C. RESULTS: Performance time is approximately 15 to 20 min. There is no need to select a specific test location and it is convenient to use even without a professional examiner. When the top and bottom 10%, which correspond to the outlier values statistically in the healthy subjects, were used as the cutoff values abnormal results were observed in approximately 25% of the liver cirrhosis patients. Moreover, 58% of the patients had abnormal values according to the results of at least one of the tests. CONCLUSION: It is expected that this test will be used to further assess the diagnosis and pathology of SHE and that it will be utilized as a routine method of diagnosis. PMID- 19125944 TI - Treatment of minimal hepatic encephalopathy. AB - Several reports, conducted as a placebo-controlled, double-blind study, presented a favorable result in treating MHE. They included branched chain aminoacid, fluzazenil-a benzodiazepin receptor antagonist, lactulose, lactitol, and L ornithin-L-aspartate. Lactulose and lactitol have been shown to be effective in MHE. It improved psychomotor tests and lowered ammonia levels as well as quality of life. Branched chain amino acids (BCAAs) were reported to improve nitrogen metabolism, blood ammonia level, and psychomotor tests. Flumazenil, an antagonist of benzodiazepine receptor, has not been associated with established consensus on the effectiveness in MHE. L-ornithine-L-aspartate (OA) exerts its ammonia lowering action in the kidney, skeletal muscles, brain, as well as in the liver. OA administered per orally improved number connection test, ammonia levels, and mental state. OA may be a promising therapy for patients with MHE. A shunt closure manipulation with balloon occluded retrograde transvenous obliteration (BRTO) has been shown to be effective for hepatic encephalopathy and its efficacy in MHE has not been elucidated. Synbiotic modulation of gut flora. was shown to increase fecal content of non-urease-producing Lactobacillus species and this change was associated with a reversal of MHE. In conclusion, there are no definitive conclusion on the treatment of MHE because of difficulties in diagnosis and evaluation. Therapeutic strategy should be planned specifically for each patient depending on the etiological factors. PMID- 19125945 TI - Clinical epidemiology of fulminant hepatitis in Japan. AB - The annual incidence of fulminant hepatitis (FH) in Japan has decreased from 3700 patients in 1972 to 1000 patients in 1989, 1050 patients in 1995 and 426 in 2004. The most frequent cause of FH in Japan is hepatitis B virus (HBV)-related hepatitis, which accounts for around 40%, with hepatitis A accounting for around 10%; drug reaction accounts for around 10%; other hepatitis, including an unknown cause, accounts for around 40%. The acute type FH, which has a disease duration as 10 days or shorter before the development of encephalopathy (onset-coma days [OCD]), mainly consists of hepatitis A and B and has relatively better prognosis, with an approximately 40% survival rate without transplantation. The subacute type FH, which has 11 to 56 days OCD, mainly consists of unknown hepatitis including acute-onset autoimmune hepatitis and has poorer prognosis of about 20% survival. Approximately 25% of FH patients underwent liver transplantation and about 80% of them survived. High volume plasma exchange (PE) and hemodialysis filtration (HDF) have been the most common and principal therapies and they are administered to about 90% and 70% of patients, respectively. The progress of artificial liver support such as combination treatment of PE with HDF is considered to play a role in the slight improvement of the survival proportion of patients with acute type FH in recent years. PMID- 19125946 TI - Implication of cytokines: Roles of tumor necrosis factor-alpha in liver injury. AB - Acute liver failure caused by viruses, drugs, or liver resection, is marked by a massive degree of hepatocyte apoptosis and impaired hepatocyte proliferation, the mechanisms of which, however, still remain to be understood. The choice between life and death is associated with events in regulation of the immune system. The liver is continuously exposed to a large antigenic load that includes pathogens, toxins and dietary antigens. Bacterial toxins, including endotoxin and staphylococcal enterotoxin, have been implicated in the pathogenesis of multi organ failure associated with liver damage through production of cytokines and chemokines. Inflammation involves the sequential activation of signaling pathways leading to the production of both pro- and anti-inflammatory mediators. Among pro inflammatory mediators, tumor necrosis factor-alpha (TNF-alpha)/TNF receptor (TNFR) systems play central roles in the physiological regulation of apoptosis as well as inflammation and immunity. These pleiotropic biological effects of TNF alpha result from its ability to initiate different intracellular signaling pathways, which induce both pro-apoptotic and anti-apoptotic molecules. Hepatocytes appear to be poorly responsive to pro-apoptotic stimuli by TNF-alpha. Tumor necrosis factor-alpha, however, induces excessive hepatocyte apoptosis, once cells are sensitized by D-galactosamine or actinomycin D, suggesting that TNF-alpha itself also induces molecules that protect cells from apoptosis by TNF alpha. Besides the apoptosis-inducing signal, the binding of TNF-alpha to TNFR1 triggers a series of intracellular events that result in the activation of nuclear factor-kappaB (NF-kappaB), phosphatidylinositol 3-kinase (PI3K)/Akt, and c-Jun NH(2)-terminal kinase (JNK). Inhibition of NF-kappaB may be a two-edged sword against liver injury, which inhibits pro-inflammatory gene expression in leukocytes and causes the sensitization of hepatocytes to TNF-alpha-induced apoptosis. A variety of mechanisms exist to modulate the activity of intracellular molecules and thereby affect the ultimate outcome of a liver cell's fate. PMID- 19125947 TI - Program. PMID- 19125948 TI - Impaired liver regeneration in acute liver failure: the significance of cross communication of growth associated factors in liver regeneration. AB - Liver regeneration is considered to be retarded or suppressed in patients with acute liver failure. However, the mechanisms for this suppression remain to be elucidated. In order to evaluate deficiencies in liver regeneration in patients with acute liver failure, we focused on the cross-communication of growth associated factors in experimental models. In primary cultured rat hepatocytes, the levels of cellular p53 and transforming growth factor-alpha (TGF-alpha), as well as DNA synthesis increased by the addition of hepatocyte growth factor (HGF). When p53 activity was suppressed, TGF-alpha expression and DNA synthesis were reduced. DNA synthesis was also reduced when TGF-alpha activity was suppressed. In rats after partial hepatectomy, hepatic HGF and p53 protein levels increased, followed by an increase of hepatic TGF-alpha levels and hepatocyte proliferation. Circulating levels of HGF and TGF-alpha correlated with changes in the hepatic levels. The suppression of TGF-alpha activity reduced the proliferation of hepatocytes in these rats. In patients suffering from acute hepatitis, serum HGF levels increased followed by an increase of serum TGF-alpha levels. In contrast, in patients with acute liver failure, the increase of serum TGF-alpha levels was suppressed depending on the severity of hepatic failure, even though serum HGF levels markedly increased. The patients' hepatic levels were consistent with serum levels. Cross-communication of growth associated factors may be important in the progression of liver regeneration, and impaired regenerative capacity in patients with acute liver failure may be attributable, at least in part, to a disruption of communication of growth-associated factors. PMID- 19125949 TI - Acetaminophen-related acute liver failure in the United States. AB - Acetaminophen overdoses are the number one cause of acute liver failure (ALF) in the United States; they account for 50% of all cases of ALF and carry a 30% mortality. Nevertheless, acetaminophen is a highly successful product with sales easily exceeding a billion dollars annually. A narcotic-containing acetaminophen preparation is the number one generic drug sold in the United States. Historically, a strong association was claimed with alcohol as a co-factor and the accidental nature of ingestions was emphasized but the association with alcohol remains a matter of debate. Recently, a very reliable assay has been developed which detects a byproduct of the toxic reaction, acetaminophen-CYS adducts, protein-acetaminophen compounds that are released from damaged hepatocytes in similar fashion to aminotransferases. The US Acute Liver Failure Study Group is a multi-center network established to gather data prospectively on all forms of acute liver failure, a rare but often fatal condition. The group has data on more than 500 acetaminophen cases and described 275 of them in detail in a recent paper. The clinical pattern of presentation is somewhat different in the US compared with the UK, since nearly half are considered to be unintentional, the result of overuse of acetaminophen-containing compounds for pain relief, without suicidal intent. Despite the relatively good prognosis for acetaminophen overdoses, 30% die once acute liver failure occurs and this is the largest cause of death in the entire ALF registry. PMID- 19125950 TI - Extracorporeal treatment of acute liver failure. AB - Acute liver failure (ALF) is a widespread problem with typically unfavorable prognosis. With the implementation of a liver support device in the clinical setting for treatment of patients with ALF, anticipated improvements include prolonging time available for spontaneous recovery and bridging to liver transplantation. Liver support could also serve to prevent systemic manifestations of ALF such as renal failure, pulmonary edema, systemic inflammatory response syndrome and cerebral edema evolving to brain death. Both non-cell based and cell based (bio-artificial) systems have been used in clinical trials. Systems with closed or open loop organization present different advantages and disadvantages; systems also differ in the membrane pore size for filtrate/dialysate exchange. Further optimization of liver assist devices is still required; when a system has proved to be successful in treating the debilitating results of ALF, the benefits will be enormous to liver failure patients. PMID- 19125951 TI - Extracorporeal albumin dialysis. AB - Patients with liver failure still present a high mortality. It can only be significantly improved by the rare resource of liver transplantation. Extracorporeal liver support devices have been developed to temporarily support liver detoxification. Artificial devices without hepatocytes ("liver dialysis" or "albumin dialysis") are already widely used in Europe. The two best-known systems, MARS and Prometheus, use a different technical approach to remove water soluble as well as albumin-bound toxins from the blood. In MARS, toxins diffuse along a concentration gradient through an albumin-impermeable membrane into a secondary circuit that is pre-filled with an albumin solution. The albumin is continuously "recycled" inside the secondary circuit by different adsorber and low flux dialysis. In contrast, Prometheus includes an albumin-permeable filter allowing separation of the albumin fraction into the secondary circuit where the albumin-bound toxins are directly removed by two adsorbers. Thereafter, high flux dialysis is performed inside the primary circuit. For both extracorporeal systems, an improvement of hepatic encephalopathy and biochemical markers such as bilirubin is consistently reported. In-vivo comparisons of both systems showed significantly higher extraction capacities for protein-bound and water-soluble substances under Prometheus than under MARS treatment. Possible pathophysiological mechanisms could be a reduction of portal pressure or a removal of vasoactive cytokines. However, only few randomised controlled trials with low patient numbers and conflictive results regarding patient survival exist. Nevertheless, a Cochrane meta-analysis revealed a significant survival benefit for extracorporeal liver support devices in patients with acute-on chronic liver failure. Other promising indications are severe refractory cholestatic pruritus, intoxication with protein-bound substances and graft dysfunction after liver transplantation. As large randomised controlled multi center trials are currently underway, better evidence will be available soon to define the clinical role of extracorporeal liver support devices. PMID- 19125952 TI - Plasma exchange in combination with online-hemodiafiltration as a promising method for purifying the blood of fulminant hepatitis patients. AB - BACKGROUND: Fulminant hepatitis is an intractable disease caused by various etiological agents. Artificial liver support (ALS) is a symptomatic treatment used to control serious symptoms, such as bleeding tendency, hepatic coma, and brain edema. METHODS: The present study involved four patients with fulminant hepatitis who were admitted to Showa University Fujigaoka Hospital between January 2007 and June 2007. All four patients were subacute type disease of indeterminate etiology. The four patients were placed on an ALS system that comprised plasma exchange and online hemodiafiltration. The effect of the ALS on various symptoms of fulminant hepatitis was evaluated, and the levels of glutamine in the patients' plasma samples and the discarded buffer were assayed using automatic analyser. RESULTS: Three of the four patients regained full consciousness and survived. The remaining patient died despite recovering from hepatic coma with ALS. The plasma glutamine levels were significantly reduced by artificial liver support. The estimated distribution volume of removed Gln ranged from 30 L to 60 L. CONCLUSIONS: Plasma exchange in combination with online hemodiafiltration is a promising and effective method for purifying the blood of patients with fulminant hepatitis. PMID- 19125953 TI - Indication criteria for liver transplantation for acute liver failure in Japan. AB - In Japan, patients with hepatitis are diagnosed as having fulminant hepatitis and late-onset hepatic failure (LOHF) when grade II or more severe hepatic encephalopathy develops within 8 weeks and between 8 and 24 weeks, respectively, of the disease onset, with a prothrombin time of less than 40% of the standardized value. Fulminant hepatitis patients are classified into the acute and subacute types, in which the encephalopathy occurs within 10 days and later than 11 days, respectively. According to the nation-wide survey by the Intractable Liver Disease Study Group of Japan, liver transplantation was performed in 14% and 26% of the patients with the acute and subacute types, respectively, and in 19% with LOHF between 1998 and 2003. Survival rate of these patients was 77%, which was greater than those of the patients treated without liver transplantation; 54%, 24% and 12% in the acute and subacute types and LOHF, respectively. The indications for liver transplantation in fulminant hepatitis patients are currently determined according to the Guideline of the Acute Liver Failure Study Group of Japan in 1996, which is based on assessment of the prognosis of the patients at the onset of hepatic encephalopathy and reassessed 5 days later. Predictive accuracy of the Guideline, assessed in the patients between 1998 and 2003, were 68% and 78% among the cases with the acute and subacute types, respectively, of fulminant hepatitis. Liver transplantation was considered in most of LOHF patients at 8 or more days before encephalopathy development. Although liver transplantation improved the prognosis of patients with acute liver failure, the Guideline should be modified to improve the accuracy for fulminant hepatitis patients, and the new criteria should be made for LOHF patients. PMID- 19125954 TI - Living donor liver transplantation for patients with fulminant hepatic failure. AB - The prognosis for patients with fulminant hepatic failure has improved since the introduction of liver transplantation. However, the death rate of patients awaiting liver transplantation is high, possibly because of the difficulty in obtaining grafts in a timely manner, given the relative shortage of cadaveric donors. Under these circumstances, living donor liver transplantation is an alternative therapeutic option for patients with fulminant hepatic failure. The present review provides recent updates on the clinical and therapeutic aspects of living donor liver transplantation for fulminant hepatic failure. PMID- 19125955 TI - Acute liver failure and living donor liver transplantation. AB - Acute liver failure (ALF) is defined by the presence of hepatic encephalopathy due to severe liver damage in patients without pre-existing liver disease. Although the mortality of ALF without liver transplantation is over 80%, the survival rates of ALF patients have improved considerably with the advent of liver transplantation, up to 60-80% in the last decade. Living donor liver transplantation (LDLT), which has mainly evolved in Asian countries where organ availability from deceased donors is extremely scarce, has also improved the survival rate of ALF patients. According to recent reports, the overall survival rate of adult ALF patients who underwent LDLT is 60% to 90%. Although there is still controversy regarding the graft type, the optimal graft volume, and ethical issues of defining the indications for LDLT in ALF patients with respect to donor risk, LDLT has become an established treatment option for ALF in areas where the use of deceased donors organs is severely restricted. PMID- 19125956 TI - Current status of autologous bone marrow cell infusion therapy for liver cirrhosis patients. AB - This article reviews recent translational research on the development of cell therapy using autologous bone marrow cell for liver cirrhosis patients. A clinical study of autologous bone marrow cell infusion (ABMI) therapy for liver cirrhosis patient was begun on Nov. 14, 2003. We previously reported nine liver cirrhosis (LC) cases that underwent ABMI from the peripheral vein and followed their progress up to 24 weeks. Subjects were LC patients with total bilirubin TB of < 3.0 mg/dL, platelets > 5 (10(10)/L) and no viable hepatocellular carcinoma on diagnostic imaging. Mononuclear cells (MNCs) were separated by cell washing and were infused via the peripheral vein. After ABMI therapy liver function and Child-Pugh Score were significantly improved at 4 weeks and 24 weeks (P < 0.05). A total of 23 cases of ABMI therapy have been performed for LC patients to date. No major adverse effects were noted due to ABMI therapy. In conclusion, ABMI therapy should be considered as a novel treatment for LC patients. PMID- 19125957 TI - Liver tissue engineering: The future of liver therapeutics. AB - The applications derived from the concept of tissue engineering have spurred significant interest in the field of regenerative medicine as novel, next generation therapies. Due to a lack of treatment modalities for patients suffering from many forms of liver diseases, recent studies have touted that engineering hepatic tissues de novo in culture may be a viable method to address this therapeutic void. Liver tissue engineering is a new and emerging field in which a functional liver system is created in vivo using isolated hepatocytes and/or other cells types to treat acute and chronic liver diseases. Under circumstances in which a small, but functional liver tissue system could be engineered to provide the equivalent biological function proportional to a few percent of a normal, well-functioning liver, it would be possible to correct many disease phenotypes as a result of various forms of inherited metabolic deficiencies. Alternatively, hepatic tissues can be engineered rapidly to produce therapeutic effects allowing this approach to become an effective modality in the treatment of acute liver failure. Strategies to achieve high levels of hepatocyte survival and the development of methods to engineer a functional liver system in vivo will be discussed in this review. PMID- 19125958 TI - Translational research on HGF: A phase I/II study of recombinant human HGF for the treatment of fulminant hepatic failure. AB - Hepatocyte growth factor (HGF) is a potential therapeutic agent for fatal liver diseases, including fulminant hepatic failure (FHF). After performing a number of preclinical tests with recombinant human HGF (rh-HGF), we started a phase I/II study in September 2005 of patients with FHF or late-onset hepatic failure (LOHF), to examine the safety and clinical efficacy of rh-HGF. We first administered rh-HGF (0.6 mg/m(2)/day) for 13 days to a 67-year-old Japanese man with FHF. All data from this patient were reviewed by the independent data monitoring committee, and the safety of rh-HGF was recognized. Finally, a clinical trial of rh-HGF was approved to be continued. As of August 2007, we have administered rh-HGF to four patients with FHF or LOHF. All patients showed a moderate decrease in systolic blood pressure during rh-HGF administration, while the urinary excretion of albumin did not increase in all cases. In the first and third patients, hepatic failure gradually progressed, and they died 66 and 29 days, respectively, after encephalopathy occurred. The second and fourth patients are presently still alive. In conclusion, we started a clinical trial that examined the effects of rh-HGF in patients with FHF or LOHF, and in the four patients with FHF or LOHF enrolled in this study, repeated doses of rh-HGF did not produce any severe side effects. PMID- 19125959 TI - Acute liver failure related to hepatitis B virus. AB - Hepatitis B virus (HBV) is an important cause of acute liver failure (ALF) worldwide. Two clinical entities are responsible for HBV-related ALF: acute hepatitis B and acute exacerbation of chronic HBV infection. Differentiation between these clinical entities is sometimes difficult, especially when past status of hepatitis B surface antigen (HBsAg) is unknown. A quantitative analysis of serum immunoglobulin M (IgM) antibody to the hepatitis core antigen (anti-HBc) may be helpful in the differential diagnosis. The recent definition of ALF in chronic HBV infection is somewhat problematic, because clinical, laboratory and radiological findings cannot reliably predict the presence of underlying liver cirrhosis in patients with acute exacerbation of chronic HBV infection. Explant liver shows underlying liver cirrhosis in >50% of these cases, even when there is no clinical evidence of liver cirrhosis. Therefore, it is advisable to exclude these patients in clinical trials because of the high incidence of underlying liver cirrhosis. The clinical course, however, is not different and neurologic complications are responsible for about half of the deaths, so a priority for liver transplantation should be maintained in these patients. The prognosis of HBV-related ALF is quite grim without liver transplantation with transplant-free survival rates ranging from 26% to 53%. To date, there is not enough evidence that antiviral therapy or artificial liver support devices can enhance the patient's survival, therefore, liver transplantation is still the only effective treatment option that can alter the grave prognosis of HBV-related ALF. PMID- 19125960 TI - Nutritional treatment of chronic liver failure. AB - The liver plays a central role in the regulation of nutrition by trafficking the metabolism of nutrients, their distribution and appropriate use. Accordingly, protein-energy malnutrition is common in patients with advanced liver disease, and it is a significant prognostic factor, affecting survival, the success of liver transplantation and quality of life. Clinical guidelines for the assessment and treatment of malnutrition have been issued by International societies, suggesting that nutritional therapy should be instituted in all patients where requirements are not adequately met by diet. The supplementation of the diet with amino acids (mainly branched-chain amino acids) and trace elements may improve nutritional status, liver function and hepatic encephalopathy. Nutritional issues should be carefully considered in Liver Units treating patients with advanced cirrhosis, and long-term, carefully controlled studies are needed to better define the type of nutritional support and the amount and timing of administration. PMID- 19125961 TI - AIDS-associated cryptococcosis: a comparison of epidemiology, clinical features and outcome in the pre- and post-HAART eras. Experience of a single centre in Italy. AB - Objectives To assess the prevalence, clinical and immunological characteristics, risk factors and survival of patients with AIDS-related cryptococcosis in the era of highly active antiretroviral therapy (HAART). Methods All newly diagnosed cryptococcosis cases identified retrospectively from among a series of AIDS patients hospitalized consecutively at a single institution in Italy in 1985-1996 (pre-HAART period, n=165) and 1997-2006 (post-HAART period, n=40) were analysed comparatively. Results The prevalence of cryptococcosis decreased from 4.7% (165/3543) to 2.2% (40/1805) between the pre- and post-HAART periods (P=0.0001). There were no differences in the clinical features or immunological status of the patients between the two cohorts. The variables associated with the occurrence of cryptococcosis in the post-HAART era were older age (P<0.001), no previous diagnosis of HIV infection (P<0.001) and infection in homosexual males (P=0.004). During the post-HAART period, immune reconstitution inflammatory syndrome associated with cryptococcosis was observed in five patients (19.3%) a median of 15 weeks after the start of HAART. Thirty-day survival (P=0.045) and overall survival (P=0.0001) were significantly better among patients diagnosed with cryptococcosis in the post-HAART compared to those diagnosed in the pre-HAART era. Conclusions The AIDS-associated cryptococcosis observed in Western countries in the HAART era has similar clinical and immunological characteristics to that observed in the pre-HAART era, but a significantly better outcome. PMID- 19125963 TI - Re: Efficacy of influenza vaccination in HIV-positive patients: a systematic review and meta-analysis. PMID- 19125962 TI - Hyperhomocysteinaemia in HIV-infected patients: determinants of variability and correlations with predictors of cardiovascular disease. AB - OBJECTIVE: We evaluated hyperhomocysteinaemia (HHcy) in a cohort of HIV-infected patients in order to assess its relation to cardiovascular risk (CVR) and identify determinants of HHcy variability. METHODS: Cross-sectional observational study. HIV-infected patients on stable highly active antiretroviral therapy (ART) were evaluated for the presence of the metabolic syndrome, lipodystrophy and traditional CVR factors. Plasma homocysteine levels were measured using high performance liquid chromatography. RESULTS: Five hundred and sixty-seven patients (38% female) with a median age of 44 years were included in the study. Homocysteine (Hcy) was significantly higher in patients with the metabolic syndrome and lipodystrophy. No significant association was found between Hcy levels and the use of ART. However, Hcy was associated with higher blood pressure, waist circumference and waist-to-hip ratio, total lean body mass, visceral adipose tissue (VAT), VAT/total adipose tissue, homeostasis model assessment of insulin resistance (HOMA-IR), triglycerides, high-density lipoprotein cholesterol, apolipoprotein A1, B, and creatinine. All 10-year CVR assessment scores were significantly associated with Hcy. In a multivariate regression model, systolic blood pressure, vitamin supplementation and HOMA-IR were significantly and independently related to Hcy. CONCLUSIONS: Hcy is elevated in HIV-infected patients and is significantly associated with increased CVR. Measurement of Hcy might be useful in identifying particularly high-risk populations at whom therapeutic interventions could be targeted. PMID- 19125965 TI - Influence of aminotransferase values on liver stiffness measurement: the case of HIV-infected patients with acute viral hepatitis B or C. PMID- 19125966 TI - Evaluation of the signalling and referral system for households at risk of eviction in Amsterdam. AB - In Amsterdam, over 1400 households are evicted each year. We describe the results of an evaluation of the functioning of the signalling and referral system, set up for households at risk of eviction, through a qualitative and quantitative study. Interviews and questionnaires completed by employees of 12 housing associations (for rent arrears) and by employees of 13 nuisance control care networks (for nuisance), were used. Data on households with rent arrears, for which a court eviction order was requested, were collected prospectively in September and October 2003, and retrospectively on households causing nuisance and/or who were known to be evicted due to nuisance in 2001-2003. Functioning of signalling, of the 'alarm' of problems underlying rent arrears and/or nuisance, was evaluated by the extent of problems that were identified by the employees. Functioning of referral was evaluated by comparing the identified problems with the assistance contacts. For 275 households with rent arrears, housing associations reported social problems in 196 (71%), of whom 94 (48%) were in contact with social assistance, and medical problems in 62 (23%) of whom 18 (29%) were in contact with medical assistance. House visits resulted in a much higher identification of problems, and were associated with a reduced eviction risk [relative risk 0.57 (95% confidence interval: 0.43-0.75)]. For 190 nuisance households, nuisance control care networks reported social problems in 103 (54%), of which 13 (13%) were in contact with social assistance, and medical problems in 155 (82%), of which 142 (92%) were in contact with medical assistance. To prevent evictions in Amsterdam, housing associations should improve their signalling role by conducting more house visits, and they should refer more households to medical assistance. Nuisance control care networks should refer more households to social assistance. Only a systematic and integrated approach can keep more households at home. PMID- 19125975 TI - Effectiveness of ozone against endodontopathogenic microorganisms in a root canal biofilm model. AB - AIM: To assess the antimicrobial efficacy of aqueous (1.25-20 microg mL(-1)) and gaseous ozone (1-53 g m(-3)) as an alternative antiseptic against endodontic pathogens in suspension and a biofilm model. METHODOLOGY: Enterococcus faecalis, Candida albicans, Peptostreptococcus micros and Pseudomonas aeruginosa were grown in planctonic culture or in mono-species biofilms in root canals for 3 weeks. Cultures were exposed to ozone, sodium hypochlorite (NaOCl; 5.25%, 2.25%), chlorhexidine digluconate (CHX; 2%), hydrogen peroxide (H(2)O(2); 3%) and phosphate buffered saline (control) for 1 min and the remaining colony forming units counted. Ozone gas was applied to the biofilms in two experimental settings, resembling canal areas either difficult (setting 1) or easy (setting 2) to reach. Time-course experiments up to 10 min were included. To compare the tested samples, data were analysed by one-way anova. RESULTS: Concentrations of gaseous ozone down to 1 g m(-3) almost and aqueous ozone down to 5 microg mL(-1) completely eliminated the suspended microorganisms as did NaOCl and CHX. Hydrogen peroxide and lower aqueous ozone concentrations were less effective. Aqueous and gaseous ozone were dose- and strain-dependently effective against the biofilm microorganisms. Total elimination was achieved by high-concentrated ozone gas (setting 2) and by NaOCl after 1 min or a lower gas concentration (4 g m(-3)) after at least 2.5 min. High-concentrated aqueous ozone (20 microg mL(-1)) and CHX almost completely eliminated the biofilm cells, whilst H(2)O(2) was less effective. CONCLUSION: High-concentrated gaseous and aqueous ozone was dose-, strain- and time-dependently effective against the tested microorganisms in suspension and the biofilm test model. PMID- 19125976 TI - Stress distribution of three NiTi rotary files under bending and torsional conditions using a mathematic analysis. AB - AIM: To compare and evaluate the stress distribution of three NiTi instruments of various cross-sectional configurations under bending or torsional condition using a finite-element analysis model. METHODOLOGY: Three NiTi files (ProFile, ProTaper and ProTaper Universal) were scanned using Micro-CT to produce a three dimensional digital model. The behaviour of the instrument under bending or torsional loads was analysed mathematically in software (ABAQUS V6.5-1), taking into consideration the nonlinear mechanical characteristic of NiTi material. RESULTS: ProFile showed the greatest flexibility, followed by ProTaper Universal and ProTaper. The highest stress was observed at the surface near the cutting edge and the base of (opposing) flutes during cantilever bending. Concentration of stresses was observed at the bottom of the flutes in ProFile and ProTaper Universal instruments in torsion. The stress was more evenly distributed over the surface of ProTaper initially, which then concentrated at the middle of the convex sides when the amount of angular deflection was increased. CONCLUSION: Incorporating a U-shaped groove in the middle of each side of the convex triangular design lowers the flexural rigidity of the origin ProTaper design. Bending leads to the highest surface stress at or near the cutting edge of the instrument. Stress concentration occurs at the bottom of the flute when the instrument is subjected to torsion. PMID- 19125977 TI - Ex vivo study on root canal instrumentation of two rotary nickel-titanium systems in comparison to stainless steel hand instruments. AB - AIM: To investigate instrumentation time, working safety and the shaping ability of two rotary nickel-titanium (NiTi) systems (Alpha System and ProTaper Universal) in comparison to stainless steel hand instruments. METHODOLOGY: A total of 45 mesial root canals of extracted human mandibular molars were selected. On the basis of the degree of curvature the matched teeth were allocated randomly into three groups of 15 teeth each. In group 1 root canals were prepared to size 30 using a standardized manual preparation technique; in group 2 and 3 rotary NiTi instruments were used following the manufacturers' instructions. Instrumentation time and procedural errors were recorded. With the aid of pre- and postoperative radiographs, apical straightening of the canal curvature was determined. Photographs of the coronal, middle and apical cross sections of the pre- and postoperative canals were taken, and superimposed using a standard software. Based on these composite images the portion of uninstrumented canal walls was evaluated. RESULTS: Active instrumentation time of the Alpha System was significantly reduced compared with ProTaper Universal and hand instrumentation (P < 0.05; anova). No instrument fractures occurred in any of the groups. The Alpha System revealed significantly less apical straightening compared with the other instruments (P < 0.05; Mann-Whitney U test). In the apical cross-sections Alpha System resulted in significantly less uninstrumented canal walls compared with stainless steel files (P < 0.05; chi-squared test). CONCLUSION: Despite the demonstrated differences between the systems, an apical straightening effect could not be prevented; areas of uninstrumented root canal wall were left in all regions using the various systems. PMID- 19125978 TI - Dislocation resistance of ProRoot Endo Sealer, a calcium silicate-based root canal sealer, from radicular dentine. AB - AIM: To examine the dislocation resistance of three root canal sealers from radicular dentine with and without immersion in a simulated body fluid (SBF), using a modified push-out test design that produced simulated canal spaces of uniform dimensions under identical cleaning and shaping conditions. METHODOLOGY: Sixty single-rooted caries-free human canine teeth were used. Standardized simulated canal spaces were created using 0.04 taper ProFile instruments along the coronal, middle and apical thirds of longitudinal tooth slabs. Following NaOCl/ethylenediamine tetra-acetic acid cleaning, the cavities were filled with ProRoot Endo Sealer, AH Plus Jet or Pulp Canal Sealer. After setting, half of the cavities were tested with a fibre-optic light-illuminated push-out testing device. The rest were immersed in SBF for 4 weeks before push-out evaluation. Failure modes were examined with stereomicroscopy and field emission (FE) scanning electron microscopy. RESULTS: Location of the sealer-filled cavities did not affect push-out strengths. ProRoot Endo Sealer exhibited higher push-out strengths than the other two sealers particularly after SBF storage (P < 0.001). Failure modes were predominantly adhesive and mixed for Pulp Canal Sealer and AH Plus Jet, and predominantly cohesive for ProRoot Endo Sealer. Spherical amorphous calcium phosphate-like phases that spontaneously transformed into apatite-like phases were seen in the fractured specimens of ProRoot Endo Sealer after SBF storage. CONCLUSIONS: When tested in bulk without a main core, both 'sealer type' and 'SBF storage' were significant in affecting push-out results. The ProRoot Endo Sealer demonstrated the presence of spherical amorphous calcium phosphate like phases and apatite-like phases (i.e. ex vivo bioactivity) after SBF storage. PMID- 19125979 TI - Influence of post fit and post length on fracture resistance. AB - AIM: To investigate (i) the impact of post fit (form-congruence) and (ii) the influence of post length on the fracture resistance of severely damaged root filled extracted teeth. METHODOLOGY: Ninety-six single-rooted human teeth were root filled and divided into four groups (n = 24 per group). Post spaces were prepared with a depth of 6 mm (group 1, 3) and 3 mm (group 2, 4). Form-congruence with a maximal fit of the post within the root canal space was obtained in groups 1 and 2, whereas there was no form-congruence in groups 3 and 4. In all groups, glass fibre reinforced composite (FRC) posts were adhesively cemented and direct composite crown build-ups were fabricated without a ferrule. After thermo mechanical loading (1200000x, 5-50 degrees C), static load was applied until failure. Loads-to-failure [in N] were compared amongst the groups. RESULTS: Post fit did not have a significant influence on fracture resistance, irrespective of the post length. Both groups with post insertion depths of 6 mm resulted in significantly higher mean failure loads (group 1, 394 N; group 3, 408 N) than the groups with post space preparation of 3 mm (group 2, 275 N; group 4, 237 N). CONCLUSIONS: Within the limitations of this study, the fracture resistance of teeth restored with FRC posts and direct resin composite crowns without ferrules was not influenced by post fit within the root canal. These results imply that excessive post space preparation aimed at producing an optimal circumferential post fit is not required to improve fracture resistance of roots. PMID- 19125980 TI - Root canal morphology of mandibular first premolars in an Indian population: a laboratory study. AB - AIM: To determine the root canal morphology of mandibular first premolar teeth in an Indian population using a decalcification and clearing technique. METHODOLOGY: One hundred extracted adult mandibular first premolar teeth were studied following decalcification and clearing. The shape of the canal orifice, root canal pattern and length of the teeth were determined. RESULTS: The mandibular first premolars were identified to have a round orifice (38%), oval orifice (44%), flattened orifice (17%) and C-shaped orifice (1%). The canal patterns were classified as Type I (72%), Type II (6%), Type III (3%), Type IV (10%) and Type V (8%) according to Vertucci's classification. C-shaped canals were identified in one tooth (1%). The average length of the teeth was 21.6 mm. Fourteen per cent of the teeth had mesial invaginations of the root. CONCLUSIONS: Type I canal patterns were the most frequently occurring in mandibular first premolars amongst the Indian population. 85.7% of the teeth with mesial invagination of the root had either two canals or division of canals. PMID- 19125981 TI - Necrotic pulp tissue dissolution by passive ultrasonic irrigation in simulated accessory canals: impact of canal location and angulation. AB - AIM: To evaluate whether passive ultrasonic irrigation (PUI) of 2.5% NaOCl would dissolve necrotic pulp tissue from simulated accessory root canals (SACs) better than passive placement of the irrigant, when temperature was equilibrated between the two treatments. METHODOLOGY: Transparent root canal models (n = 6) were made from epoxy resin. SACs of 0.2 mm diameter were placed at defined angles and positions in the mid-canal and apical area. SACs were filled with necrotic bovine pulp tissue. PUI was performed five times for 1 min each with irrigant replenishment after every minute. Main canal temperature was measured after each minute, and a digital photograph was taken. In control experiments, mock treatments were performed with the same set-up without activation of the file using heated NaOCl to mimic the temperature created by PUI. Experiments were repeated five times. Digital photographs were analysed for the distance of dissolved tissue into the SACs in mm. Overall comparison (sum of dissolved tissue from all five accessory canals) between treatments was performed using paired t test. Differences between SAC angulation and position after PUI were investigated using anova/Bonferroni (alpha < 0.05). RESULTS: Passive ultrasonic irrigation caused a rise in irrigant temperature in the main canal to 53.5 +/- 2.7 degrees C after the fifth minute. PUI dissolved a total of 6.4 +/- 2.1 mm, mock treatment controlled for heat: 1.4 +/- 0.6 mm (P < 0.05). No significant influence of SAC position or angulation was found. CONCLUSIONS: Passive ultrasonic irrigation promotes positive tissue-dissolving effects beyond a rise in irrigant temperature. PMID- 19125984 TI - ENDURANCE: getting the timing right. PMID- 19125982 TI - Pulp revascularization of necrotic bilateral bicuspids using a modified novel technique to eliminate potential coronal discolouration: a case report. AB - AIM: To present a case report in which the pulp of two bilateral mandibular premolars with dens evaginatus were revascularized using a modified novel technique to avoid undesired crown discolouration. SUMMARY: Recently, regeneration of necrotic pulps has become an alternative conservative treatment option for young permanent teeth with immature roots and is a subject of great interest in the field of endodontics. This novel procedure exploits the full potential of the pulp for dentine deposition and produces a stronger mature root that is better able to withstand the forces than can result in fracture. However, the current protocol has potential clinical and biological complications. Amongst them, crown discolouration, development of resistant bacterial strains and allergic reaction to the intracanal medication. In the case presented, a modified technique to avoid undesired crown discolouration was applied sealing the dentinal tubules of the chamber, thus avoiding any contact between the tri antibiotic paste and the dentinal walls. KEY LEARNING POINTS: * Sealing the dentinal tubules of the chamber prevents the undesirable crown discolouration produced by tri-antibiotic medication whilst maintaining the revascularization potential of the pulp. * Further research is warranted to seek an alternative infection control protocol capable of preventing possible allergic reactions and development of resistant strains of bacteria, as well as a biological material capable of inducing angiogenesis and allow a more predictable scaffold and tissue regeneration. PMID- 19125985 TI - Detecting peripheral arterial disease using the ankle-brachial index. PMID- 19125986 TI - Resurrecting an old drug--a flash in the pan or here to stay? PMID- 19125987 TI - Hypertension strategies in the third millennium: conservatism, evidence and the folly of speculation. PMID- 19125988 TI - Targeting triglycerides in secondary prevention: should we bother? PMID- 19125989 TI - Evidence-based guidelines for the use of biochemical markers of bone turnover in the selection and monitoring of bisphosphonate treatment in osteoporosis: a consensus document of the Belgian Bone Club. AB - OBJECTIVES: To review the clinical value of bone turnover markers (BTM), to initiate and/or monitor anti-resorptive treatment for osteoporosis compared with bone mineral density (BMD) and to evaluate suitable BTM and changes in BTM levels for significance of treatment efficiency. METHODOLOGY: Consensus meeting generating guidelines for clinical practice after review and discussion of the randomised controlled trials or meta-analyses on the management of osteoporosis in postmenopausal women. RESULTS: Although the correlation between BMD and BTM is statistically significant, BTM cannot be used as predictive markers of BMD in an individual patient. Both are independent predictors of fracture risk, but BTM can only be used as an additional risk factor in the decision to treat. Current data do not support the use of BTM to select the optimal treatment. However, they can be used to monitor treatment efficiency before BMD changes can be evaluated. Early changes in BTM can be used to measure the clinical efficacy of an anti resorptive treatment and to reinforce patient compliance. DISCUSSION: Determining a threshold of BTM reflecting an optimal long-term effect is not obvious. The objective should be the return to the premenopausal range and/or a decrease at least equal to the least significant change (30%). Preanalytical and analytical variability of BTM is an important limitation to their use. Serum C-terminal cross-linked telopeptide of type I collagen (CTX), procollagen 1 N terminal extension peptide and bone specific alkaline phosphatase (BSALP) appear to be the most suitable. CONCLUSION: Consensus regarding the use of BTM resulted in guidelines for clinical practice. BMD determines the indication to treat osteoporosis. BTM reflect treatment efficiency and can be used to motivate patients to persist with their medication. PMID- 19125991 TI - Long-term desmopressin response in primary nocturnal enuresis: open-label, multinational study. AB - BACKGROUND: Primary nocturnal enuresis (PNE) is a distressing condition, particularly in severe cases (> or = 3 wet nights/week). A prevalent pathophysiological mechanism, especially in monosymptomatic PNE (PMNE), is commonly believed to be an insufficient increase in night-time release of antidiuretic hormone. Desmopressin, a synthetic analogue of antidiuretic hormone, has been shown to reduce the number of wet nights experienced by PMNE patients in several controlled trials. AIM: This study was performed to evaluate desmopressin treatment in the real-life clinical setting and was a large-scale, 6-month investigation of efficacy and safety in patients with severe PNE. Predictive factors for desmopressin response were also evaluated. A total of 744 children aged 5 years and above from four countries were involved in the study. RESULTS: At baseline, patients had a median of 6 wet nights/week; at 6 months, 41% of patients had experienced > or = 50% reduction in the mean number of wet nights. Long-term desmopressin treatment was consistently well-tolerated across all ages, with 5% of patients experiencing any treatment-related adverse events. The strength of treatment response was associated with nocturnal diuresis (p < 0.0001) and age (p = 0.0167) in logistic regression analyses. Compliance and dosage were also associated with response and more patients experienced > or = 50% reduction in wet nights after 6 months' treatment than earlier in the study, suggesting the value of persistent treatment. CONCLUSION: This study shows that long-term desmopressin treatment in the clinical setting is effective and well tolerated in PNE patients of 5 years and upwards. Early improvements in bedwetting of any appreciable magnitude may be rewarding, may facilitate compliance and enable good long-term response. PMID- 19125990 TI - Improvement in duration of erection following phosphodiesterase type 5 inhibitor therapy with vardenafil in men with erectile dysfunction: the ENDURANCE study. AB - OBJECTIVE: The ENDURANCE study evaluated the efficacy of vardenafil, a phosphodiesterase type 5 (PDE5) inhibitor, in men with erectile dysfunction (ED), by measuring the duration of erection leading to successful intercourse using a stopwatch as the assessment instrument. METHODS: This was a randomised, multicentre, double-blind, placebo-controlled, crossover study consisting of a 4 week treatment-free run-in phase after which patients were randomised to either fixed-dose vardenafil 10 mg or placebo (to be administered 60 min prior to intercourse) and entered the first of the two 4-week double-blind treatment periods, separated by a 1-week washout. The primary efficacy end-point was the stopwatch-assessed duration of erection, which was defined as the time from erection perceived hard enough for penetration until withdrawal from the partner's vagina leading to successful intercourse as measured by Sexual Encounter Profile Question 3 (SEP-3). Secondary efficacy end-points included SEP 2 and SEP-3 success rates, the erectile function domain of the International Index of Erectile Function, global assessment questionnaire, change from baseline in duration of erection and duration of erection not leading to successful intercourse. Safety was assessed by adverse events (AEs), laboratory samples, vital signs and ECGs. RESULTS: Of the 191 men included in the safety population, 40% had moderate ED and 33% had severe ED at baseline. The duration of erection (least squares mean +/- SE) leading to successful intercourse was longer with vardenafil than with placebo (12.81 +/- 1.00 min vs. 5.45 +/- 1.00 min; p < 0.001). The differences recorded for all secondary end-points were statistically significant in favour of vardenafil compared with placebo (p < 0.001), with the exception of duration of erection not leading to successful intercourse. Vardenafil was well tolerated in this study; the majority of AEs being mild-to moderate in intensity. CONCLUSION: Vardenafil 10-mg therapy provided a statistically superior duration of erection leading to successful intercourse in men with ED compared with placebo. PMID- 19125992 TI - Efficacy and safety of adding the dipeptidyl peptidase-4 inhibitor alogliptin to metformin therapy in patients with type 2 diabetes inadequately controlled with metformin monotherapy: a multicentre, randomised, double-blind, placebo controlled study. AB - AIMS: To evaluate the efficacy and safety of alogliptin, a new dipeptidyl peptidase-4 inhibitor, for 26 weeks at once-daily doses of 12.5 and 25 mg in combination with metformin in patients whose HbA(1c) levels were inadequately controlled on metformin alone. METHODS AND PATIENTS: Patients with type 2 diabetes and inadequate glycaemic control (HbA(1c) 7.0-10.0%) were randomised to continue a stable daily metformin dose regimen (> or = 1500 mg) plus the addition of placebo (n = 104) or alogliptin at once-daily doses of 12.5 (n = 213) or 25 mg (n = 210). HbA(1c), insulin, proinsulin, C-peptide and fasting plasma glucose (FPG) concentrations were determined over a period of 26 weeks. RESULTS: Alogliptin at either dose produced least squares mean (SE) decreases from baseline in HbA(1c) of -0.6 (0.1)% and in FPG of -17.0 (2.5) mg/dl [-1.0 (0.1) mmol/l], decreases that were significantly (p < 0.001) greater than those observed with placebo. The between treatment differences (alogliptin - placebo) in FPG reached statistical significance (p < 0.001) as early as week 1 and persisted for the duration of the study. Overall, adverse events (AEs) observed with alogliptin were not substantially different from those observed with placebo. This includes low event rates for gastrointestinal side effects and hypoglycaemic episodes. There was no dose-related pattern of AE reporting between alogliptin groups and few serious AEs were reported. CONCLUSION: Alogliptin is an effective and safe treatment for type 2 diabetes when added to metformin for patients not sufficiently controlled on metformin monotherapy. PMID- 19125993 TI - Atherogenic forms of dyslipidaemia in women with polycystic ovary syndrome. AB - OBJECTIVE: Dyslipidaemia is very common in patients with polycystic ovary syndrome (PCOS) but, beyond plasma lipids, atherogenic lipoprotein (Lp) and apolipoprotein (apo) alterations are still ill defined. DESIGN: We measured concentrations of apoB, Lp(a) and small, dense low-density lipoprotein (LDL) in 42 patients with PCOS [age: 28 +/- 7 years, body mass index (BMI): 27 +/- 5 kg/m(2)] vs. 37 age- and BMI-matched healthy controls. METHODS: Elevated Lp(a) levels considered were those > 30 mg/dl while elevated apoB concentrations were those > 100 g/l. RESULTS: Polycystic ovary syndrome showed increased triglycerides levels (p = 0.0011) and lower high-density lipoprotein (HDL) cholesterol concentrations (p = 0.0131) while total- and LDL cholesterol were similar. PCOS also showed smaller LDL size (p = 0.0005), higher levels of total small, dense LDL (p < 0.0001), higher concentrations of Lp(a), as considered as absolute values (p = 0.0143) and log-transformed (p = 0.0014), while no differences were found in apoB levels. Elevated Lp(a) concentrations were found in 24% of PCOS, while elevated apoB levels were relatively uncommon (14%). Spearman correlation analysis revealed that Lp(a) concentrations were weakly correlated only with HDL-cholesterol levels (r = -0.378, p = 0.0431). In addition, 36% of patients with PCOS with normal plasma lipid profile showed elevated levels of Lp(a), apoB or small, dense LDL. CONCLUSIONS: Atherogenic Lp abnormalities may be found in one-third of women with PCOS who have a normal lipid pattern. Future prospective studies are needed to test to which extent such atherogenic forms of dyslipidaemia may contribute to the increased cardiovascular risk in young women with PCOS. PMID- 19125994 TI - Prevalence of peripheral arterial disease in high-risk patients using ankle brachial index in general practice: a cross-sectional study. AB - AIMS: The deleterious nature of peripheral arterial disease (PAD) is compounded by a status of underdiagnosed and undertreated disease. We evaluated the prevalence and predictive factors of PAD in high-risk patients using the ankle brachial index (ABI). METHODS: The ABI was measured by general practitioners in France (May 2005-February 2006) in 5679 adults aged 55 years or older and considered at high risk. The primary outcome was prevalence of PAD (ABI strictly below 0.90). RESULTS: In all, 21.3% patients had signs or symptoms suggestive of PAD, 42.1% had previous history of atherothrombotic disease and 36.6% had two or more cardiovascular risk factors. Prevalence of PAD was 27.8% overall, ranging from 10.4% in patients with cardiovascular risk factors only to approximately 38% in each other subgroup. Prevalence differed depending on the localization of atherothrombotic events: it was 57.1-75.0% in patients with past history of symptomatic PAD; 24.6-31.1% in those who had experienced cerebrovascular and/or coronary events. Regarding the classical cardiovascular risk factors, PAD was more frequent when smoking and hypercholesterolemia history were reported. PAD prevalence was also higher in patients with history of abdominal aortic aneurysm, renal hypertension or atherothrombotic event. Intermittent claudication, lack of one pulse in the lower limbs, smoking, diabetes and renovascular hypertension were the main factors predictive of low ABI. CONCLUSIONS: Given the elevated prevalence of PAD in high-risk patients and easiness of diagnosis using ABI in primary care, undoubtedly better awareness would help preserve individual cardiovascular health and achieve public health goals. PMID- 19125995 TI - Prevalence of low HDL cholesterol, and relationship between serum HDL and cardiovascular disease in elderly Spanish population: the PREV-ICTUS study. AB - OBJECTIVE: To assess the prevalence of low serum high-density lipoprotein cholesterol (HDL-C) concentration and the relationship between HDL-C and established cardiovascular disease (CVD) in an elderly Mediterranean population. METHODS: Analysis of Prevencion del Riesgo de Ictus, a population-based study on Spanish subjects aged > or = 60 years. Low HDL-C was defined following the European guidelines for cardiovascular prevention [men: < 40 mg/dl (< 1.0 mmol/l); women: < 46 mg/dl (< 1.2 mmol/l)]. The relationship between low HDL-C or HDL-C concentration (in quintiles) and CVD was assessed through multivariate models that included cardiovascular risk factors, statins and subclinical organ damage. RESULTS: On 6010 subjects (71.7 years, 53.5% women), low HDL-C was present in 17.5% [95% confidence interval (CI): 16.5-18.5] and was more frequent in women [20.4% (19.0-21.8) vs. 14.1% (12.8-15.4) in men p < 0.001] and in patients with diabetes, CVD or statin therapy. Low HDL-C was independently associated with CVD [adjusted odds ratio (OR): 1.46, 95% CI: 1.22-1.74, p < 0.001]. The prevalence of CVD was higher as HDL-C concentration was lower (chi square trend < 0.001). Compared with the highest quintile [> 65 mg/dl (> 1.67 mmol/l)], adjusted OR for CVD were 1.39 (1.10-1.76), 1.41 (1.11-1.80), 1.49 (1.18 1.89) and 1.91 (1.52-2.39), respectively for those in the fourth [57-65 mg/dl (1.46-1.67 mmol/l)], third [51-56 mg/dl (1.31-1.45 mmol/l)], second [46-50 mg/dl (1.18-1.30 mmol/l)] and first [< 46 mg/dl (< 1.18 mmol/l)] quintiles of HDL-C. This association was seen in males and females. CONCLUSIONS: A total of 17.5% of this Spanish population aged > or = 60 years had low HDL-C. We found a strong, independent and inverse association between HDL-C concentrations and established CVD, even at ranges of HDL-C considered as normal. PMID- 19125996 TI - Study of the burden on patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Health-related quality of life measures are widely used in patients with chronic obstructive pulmonary disease (COPD). However, they are extremely limited when used to evaluate patients outside the clinical trials. The aim of this study was to analyse the burden of the disease using a simple, validated, self-administered questionnaire specifically developed for patients in daily clinical practice. METHODS: A total of 3935 patients (74.5% men; mean age, 67 years) participated in a cross-sectional study. The burden of COPD on patients was measured using the Clinical COPD Questionnaire (CCQ). COPD was rated at four levels by the forced expiratory volume in one second (FEV(1)) according to The Global Initiative for Chronic Obstructive Lung Disease (GOLD) scale. RESULTS: The disease mainly affects old men (more than 50% were over 65 years of age) and non employed men (23% were employed). Of the patients studied, 22.7% continued smoking, especially men (24.4% of men vs. 18.1% of women). Most patients (54%) were diagnosed with moderate stage II COPD. Severity of COPD was lower in women: 29.6% of men had severe COPD compared with 13.7% of women. During the last year, 65.1% had at least one acute exacerbation and 36.6% were admitted to hospital because of COPD exacerbation. No association was found between the body mass index and COPD stage. The variable that most influenced the disease burden was dyspnoea, as progression from grade 0 to grade 4 increased the disease burden by 1.78 points for symptoms, 2.43 for functional state and 1.53 for mental state. The functional classification of COPD also had a significant influence on the disease burden. CONCLUSIONS: The present findings show that dyspnoea and the degree of airflow limitation are the clinical variables that most affect the burden of COPD from the patient's point of view. PMID- 19125997 TI - Is reliance on self-medication and pharmacy care adequate for rhinitis patients? AB - INTRODUCTION: Many people with rhinitis choose to self-medicate, frequently relying on pharmacy advice. We have investigated the effectiveness of such a treatment. METHODS: To obtain data regarding treatment and outcome of individuals with rhinitis who have not consulted a physician in the past 2 years, a cross sectional survey was performed. A random sample of the French, German and United Kingdom general population (n = 15,085) was screened, resulting in 818 individuals with rhinitis. Each of these respondents was sent a postal questionnaire. RESULTS: A total of 610 (74.6%) individuals returned the questionnaire. A sum of 42% (256) respondents described their symptoms as moderate and 5% (31) as severe. Multiple triggers were reported by 442 (72.5%) and 30% of respondents reported sleep disturbance (increased to 44% in sedating antihistamine users) > or = 8 nights/month, resulting in reduced cognition and productivity loss in 14.5%. Individuals stated that they did not attend the physician because their symptoms were too trivial (378 - 63.9%) or over-the counter (OTC) medication was adequate (341 - 57.6%). Only 217 respondents reported currently taking OTC medication (France 30; Germany 82; UK 105). In all countries, antihistamine was the main treatment taken (non-sedating in 38% and sedating in 23% of subjects). Intranasal corticosteroid use (18 - 8.3%) was mainly in the UK where it is available OTC. CONCLUSION: Individuals with rhinitis who do not consult a physician report multiple sensitization, reduced sleep and cognition. Drug utilization appears suboptimal, particularly for those reporting moderate or severe symptoms. Reliance on pharmacy or self-medication may not be adequate for some people. PMID- 19125998 TI - Efficacy and tolerability of ready-to-use intravenous paracetamol solution as monotherapy or as an adjunct analgesic therapy for postoperative pain in patients undergoing elective ambulatory surgery: open, prospective study. AB - AIMS: Paracetamol (acetaminophen) is one of the most widely used drugs for analgesia. We aimed to investigate the use of a ready-to-use intravenous (i.v.) paracetamol 1 g solution (Perfalgan) as monotherapy or as adjunct therapy in patients undergoing elective ambulatory surgery. METHODS: Open, non-controlled, observational study in six centres. Anaesthesiologists applied paracetamol 1 g intravenously about 30 min before the planned end of surgery and followed the patients up who reported postoperative pain visual analogue scale (VAS 0 mm minimum, 100 mm maximum) for pain rating until discharge (mean 123 +/- 58 min). RESULTS: A total of 601 patients (58.7% female patients, mean age 46.7 +/- 15.4 years; 54% and 42% in American Society of Anesthesiologists ASA class I or II respectively) undergoing minor knee surgery (71.4%), minor gynaecological procedures (19.0%) or varicose vein surgery (9.6%) were included, of whom 590 patients received one i.v. infusion. Mean duration of surgery was 37 +/- 21 min. Analgesic concomitant medication was applied in 57%. Mean self-reported pain intensity on the VAS was 33.2 at 15 min after end of surgery and was reduced to 19.2 at patient discharge (-13.9 points). Relative pain reduction was similar in the three surgery subtypes. The majority of patients achieved a VAS score < 30 mm and were classified as responders; i.v. paracetamol was well tolerated and no serious adverse events and only one possibly drug-related adverse event was reported. The majority of physicians (80.5%) and patients (81.6%) rated the efficacy, and satisfaction with therapy respectively, as very good or good. CONCLUSIONS: Ready-to-use i.v. paracetamol, used as monotherapy or in combination with other analgesics, may be effective for alleviating postoperative pain and well tolerated in patients undergoing ambulatory surgery. PMID- 19125999 TI - Requesting patterns for serum calcium concentration in patients on long-term lithium therapy. AB - AIM: Long-term lithium therapy is associated with hypercalcaemia in 10-60% of patients, but unlike creatinine and thyroid stimulating hormone (TSH), monitoring by general practitioners of serum calcium for patients on lithium is not a requirement of the Qualities and Outcomes Framework (QOF) of 2004. We aimed to assess requesting patterns for serum calcium in patients on long-term lithium therapy and subsequent diagnosis of hypercalcaemia. METHODS: We identified 100 patients on long-term lithium therapy, as indicated by regular monitoring of lithium levels in our laboratory for at least 1 year. We determined how many of these patients had had serum calcium analysed, noting the assay date, concentration, source of request and clinical details stated. RESULTS: Forty three out of hundred patients had serum calcium analysed during the course of their treatment including 28 in the previous 15 months. Twenty-one patients had serum calcium analysed by their GP, including 12 in the previous 15 months. Hypercalcaemia was diagnosed in five patients (11.6%). CONCLUSION: A significant proportion of patients in whom calcium was checked developed hypercalcaemia on lithium therapy. However, only 12% of the patients had serum calcium requested by their GP in the previous 15 months, which compares unfavourably with TSH and creatinine, for which monitoring approaches 100%. We recommend that serum calcium be checked every 15 months along with creatinine and TSH. This might be achieved by incorporating appropriate targets into the QOF, or by reflective or reflex adding-on of calcium to lithium specimens from patients who have not had calcium analysed in the previous 15 months. PMID- 19126000 TI - Reminders: a simple measure to ensure the mandatory venous thromboembolism risk assessment of hospitalised patients. PMID- 19126001 TI - Testosterone and cardiovascular disease: the conflicting relationship. PMID- 19126002 TI - Colo-anal intussusception in an adult: case report and review of the literature. PMID- 19126003 TI - Testicular torsion following a skin graft for Fournier's gangrene. PMID- 19126005 TI - Recurrent hard nodule of the nape of the neck. PMID- 19126006 TI - Protein folding and cutaneous diseases. PMID- 19126007 TI - p53 labeling index in assessing the efficacy of a sunscreen in protection against UV-induced damage. AB - BACKGROUND: Sunscreen efficacy is currently evaluated by the estimation of the sun protection factor (SPF). Our objective was to determine the role of the p53 labeling index in assessing the effectiveness of a sunscreen in the prevention of UV-induced DNA damage. METHODS: A broad-spectrum sunscreen (SPF15) was applied to two of four test areas on the backs of 20 volunteers. Two test areas (with and without prior sunscreen application) were then irradiated with two minimal erythemal doses (MEDs) of solar-simulated UV radiation (290-400 nm). At 24 h, suction blisters were raised from all four test areas using a vacuum technique, and immunohistochemical staining for p53 was performed. RESULTS: In normal epidermis, nonirradiated without sunscreen, p53 was almost undetectable [0.5% positive cells per field (PC/F)]. In UV-exposed skin without sunscreen, p53 expression was 10.4% PC/F, whereas, in skin treated with sunscreen before UV irradiation, p53 expression was 0.3% PC/F. The mean positive labeling indices (x10(3)/microm) for UV-exposed skin with and without sunscreen were 0.002 and 0.059, respectively (P < 0.001). CONCLUSIONS: In vivo assessment of the p53 labeling index using the suction blister method can serve as a sensitive endpoint in the evaluation of the effectiveness of a sunscreen in the prevention of UV induced DNA damage. PMID- 19126008 TI - Density of dendritic cells around basal cell carcinomas is related to tumor size, anatomical site and stromal characteristics, and might be responsible for the response to topical therapy. AB - BACKGROUND: The dermal dendritic cell (DC) is considered to be an important component of the host defense against basal cell carcinomas (BCCs). Imiquimod, an immunologic response modifier, has recently been introduced in the topical therapy of BCCs. There is some evidence that the DC pretreatment density may affect the efficacy of imiquimod. The aim of our study was to find out which clinical or histological variables are related to the DC density at the margins of BCCs. METHODS: Thirty cases of BCCs of aggressive and 30 cases of nonaggressive subtypes were selected from our files. In histological sections, the density of FXIIIa-positive DCs and 1A4-positive myofibroblasts in the tumor surrounding stroma was quantified, as well as the stroma type, the tumor size and the DC density in the normal dermis. RESULTS: In nonaggressive BBCs, a multiple linear regression showed that a higher DC density was associated with an increased number of myofibroblasts, smaller tumors and those located on the face. For the aggressive subtypes, a higher DC density was related not only to an increased myofibroblast density, smaller BCCs or location on the face, but also to the presence of less mucinous and more granulation type stroma and an increased DC density in the normal dermis. The stability of the models was confirmed by bootstrap resampling. CONCLUSION: Our study demonstrates that the density of DCs around BCCs is related to tumor size, localization and characteristics of the surrounding tumor stroma. PMID- 19126009 TI - Enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence testing in a bullous pemphigoid and pemphigoid gestationis. AB - Enzyme-linked immunosorbent assay (ELISA) is an excellent tool for detection of circulating antibodies against the NC16A portion of BP180 antigen. We compared the sensitivity and specificity of a commercially available BP180-NC16a domain ELISA with that of an indirect immunofluorescence (IIF) testing in the evaluation of bullous pemphigoid (BP) and pemphigoid gestationis (PG), and analyzed the relationship between ELISA results and the presence of IgG deposition, in an epidermal or combined pattern, on direct immunofluorescence (DIF) testing of salt split skin. ELISA was performed on serum from 28 patients (24 BP, 4 PG) and 50 controls. IIF testing was performed on serum from 27 patients and 98 controls. For the group of 28 patients with BP or PG, ELISA had a sensitivity of 93% and specificity of 96% (P < 0.001), while sensitivity was 74% and specificity 96% (P < 0.001) for IIF testing. In these patients, ELISA has a higher sensitivity than IIF testing, but similar specificity. Evaluation of controls who had IgG deposition on the dermal side of salt-split skin on DIF testing showed specificity for the ELISA of 100% (all four cases negative) and 80% for IIF testing (one of five positive). Positive ELISA correlated with a diagnosis of BP or PG only in patients who had IgG at the basement membrane zone (BMZ) by DIF testing. Overall, ELISA appears to have greater sensitivity and specificity for BP or PG than does IIF testing. PMID- 19126010 TI - Lack of evidence for hepatitis C virus infection in association with lichen planus. AB - BACKGROUND: The association between hepatitis C virus (HCV) infection and lichen planus (LP) is a subject of controversy. Prevalence studies of HCV infection in LP patients in various countries reveal diverse results. The Slovenian population is rather homogenous with specific geographic and epidemiological characteristics. Lack of data or contradictory results from neighboring countries urged the need for a case-controlled study in our LP patients. METHODS: The retrospective study was performed on 173 LP patients. Control group included 218 patients with dermatological diseases other than LP. RESULTS: Anti-HCV antibodies were found in 2/173 patients (1.2%) with LP and in 0/218 controls. No statistically significant difference was found between the study and control group regarding anti-HCV antibody prevalence (P = 0.195; estimated OR 6.4, 95% CI 0.3-134.0) and risk factors for HCV infection. CONCLUSIONS: Based on our results, anti-HCV antibody testing is not necessarily required in LP patients with no risk factors for HCV infection in this geographic region. PMID- 19126011 TI - Chanarin-Dorfman syndrome with eccrine gland vacuolation: a case report. AB - Chanarin-Dorfman syndrome is a rare congenital disorder of lipid metabolism characterized by ichthyosis, leukocytic vacuolation (Jordan's anomaly), and variable involvement of the liver and neuromuscular system, with about 40 cases described worldwide to date. We report one more case of this rare syndrome, with certain peculiarities, namely vacuolation in eccrine glands, in a young male adult. PMID- 19126012 TI - Adams Oliver syndrome--a variant. PMID- 19126013 TI - Diffuse cutaneous leishmaniasis associated with the immune reconstitution inflammatory syndrome. AB - Leishmaniasis is an emerging disease in HIV-infected persons; visceral leishmaniasis is an AIDS-defining opportunistic infection. The parasite that causes this infection is usually transmitted by the sandfly and occasionally by nonsterile needles among intravenous drug users. Diffuse cutaneous leishmaniasis (DCL) is a rare anergic variant of leishmanial infection with the characteristic presentation of numerous nonulcerating nodules with an abundant parasite load, lack of visceral involvement, negative reaction to the leishmanin skin test, and a chronic course with incomplete response to treatment and frequent relapses. We report a case of DCL that developed in the context of the immune reconstitution inflammatory syndrome (IRIS) in a man with AIDS following initiation of antiretroviral therapy. We also review DCL to emphasize the importance of recognizing and treating this evolving disease in the growing population of patients on immunorestorative therapy. PMID- 19126014 TI - Primary cutaneous actinomycosis. AB - Actinomycosis is a chronic, progressive, suppurative infection, prevalent over the neck, thorax or abdomen. As it rarely occurs with few clinical symptoms, its diagnosis requires a high index of clinical suspicion. We report a case of primary cutaneous actinomycosis affecting the back. The patient was treated with benzyl penicillin and sulfonamides and showed no recurrence at 1 year follow-up. PMID- 19126015 TI - Solitary mastocytoma: tooth eruption as triggering factor. PMID- 19126016 TI - Molluscoid skin lesions in histoid leprosy with pseudo-isomorphic Koebner phenomenon. PMID- 19126017 TI - Syphilitic mucous patches: the resurgence of an old classic. PMID- 19126018 TI - Familial nevus flammeus associated with early onset cherry angiomas. PMID- 19126019 TI - The wrinkles soothing effect on the middle and lower face by intradermal injection of botulinum toxin type A. AB - BACKGROUND: Botulinum toxin type A (BTA) has been used on facial expressive muscles to reduce wrinkles. We assumed that intradermal injection with BTA on the middle and lower face can produce face-lifting and wrinkles-soothing effect by promoting collagen synthesis. METHODS: Nine volunteers were included to undergo intradermal injections of a total dose of 20~25 U BTA into one-half of the face, and normal saline into the other half as control. Clinical photographs were taken every 4 weeks for 16 weeks, and evaluated by a doctor blinded to the assignments. Patients graded the results of wrinkles elimination on each follow-up. Skin biopsies were taken before the treatment and 8 weeks after the injection. The specimens were assessed by a pathologist using hematoxylin and eosin stain, elastin stain, Masson trichrome stain and immunohistochemical stain with type I procollagen. RESULTS: By photographic documentation, there was no significant face-lifting effect. However, there was statistical significance in wrinkles reduction on the BTA sides compared to pre-treatment. Subjectively, six subjects noticed better wrinkles soothing effect on the BTA sides. This effect was noted as early as 4 weeks after injection, and lasted for a minimum 8 weeks. The histologic examination revealed slight neocollagen synthesis by Masson trichrome stain on both sides. CONCLUSIONS: Based on this pilot study, the face-lifting effect of intradermal injection with BTA was not conclusive. Interestingly, BTA showed moderate but significant wrinkles-soothing effect without obvious side effects on the lower face. PMID- 19126020 TI - An unusual and serious complication of topical wart treatment with monochloroacetic acid. AB - Numerous chemical agents are used in the topical treatment of warts. Monochloroacetic acid (MCAA) is one of these agents, which is used in low or high concentrations in most European countries. MCAA is a strong organic acid which is irritating and corrosive to the skin and has a high systemic toxicity. In addition to wart treatment, it is used for industrial purposes, such as the synthesis of certain organic chemicals. We present a case of joint deformity manifesting after the use of a preparation containing MCAA for topical wart treatment. This underlines the need to reassess the safety of MCAA use for topical wart treatment. PMID- 19126021 TI - Hair growth stimulatory effect by a combination of 5-aminolevulinic acid and iron ion. AB - BACKGROUND: 5-Aminolevulinic acid (5-ALA) is a precursor of a tetrapyrrole compound. 5-ALA has been used for photodynamic therapy as well as for plant growth. 5-ALA and iron ion are precursors of heme, which is incorporated into hemoglobin and cytochrome. AIM: To explore the possible application of a 5-ALA and iron ion admixture on hair growth in mice. METHODS: The effect of a 5-ALA and iron ion admixture on hair growth and cell proliferation in mice was examined. The dorsal hair of 8-week-old male CeH/HeN mice was clipped, and a 5-ALA and iron ion admixture was applied to the dorsal skin once daily for 21 days in a room supplied with common room lights. Hair growth was later examined by calculating the ratio of the area showing hair growth to the total clipped area. For the cell proliferation assay, a 5-ALA and iron ion admixture at several different concentrations was added to a culture medium containing keratinocytes or fibroblasts, and the cell numbers were counted. RESULTS: Mice treated with an admixture of 5-ALA and iron ion showed significant hair growth (P < 0.05) at day 15 relative to those treated with iron ion alone, as revealed by the Tukey-Kramer test. The stimulatory effect of the mixture was almost identical to that of 5% minoxidil. No proliferation of keratinocytes or fibroblasts was observed, however, when an admixture of 5-ALA and iron ion was added to the medium. CONCLUSIONS: The results suggest that an admixture of 5-ALA and iron ion stimulates murine hair growth in vivo independent of epithelial and mesenchymal cells, although the precise mechanism is still uncertain. This mixture has the potential to become a beneficial new treatment for alopecia. PMID- 19126023 TI - Aluminum alters sweating by constricting the dermal duct lumen. PMID- 19126022 TI - Wiskott-Aldrich syndrome and molluscum contagiosum: a therapeutic challenge after reaction to cidofovir. PMID- 19126024 TI - Malignant transformation of penile lichen sclerosus: exactly how common is it? PMID- 19126025 TI - Intertriginous drug eruption: report of a case and proposed pathogenetic mechanism. PMID- 19126026 TI - Atypical skin metastases from early tubular breast carcinoma. PMID- 19126027 TI - Acute generalized exanthematous pustulosis possibly induced by acarbose. PMID- 19126029 TI - Treatment of refractory pemphigus erythematosus with rituximab. PMID- 19126028 TI - Bullous systemic lupus erythematosus induced by the therapy for lupus nephritis. PMID- 19126030 TI - Molluscum contagiosum on a tattoo: usefulness of dermoscopy. PMID- 19126031 TI - Photodynamic therapy for the treatment of in situ squamous cell carcinoma of the left eyelid. PMID- 19126032 TI - Immunoserological analyses of 55 patients with pemphigus at the Dermatological Department of Kurume University Hospital: an 11-year retrospective study (1996 2006). PMID- 19126033 TI - Recurrent subcutaneous phaeohyphomycosis caused by Exophiala sp. associated with squamocellular carcinoma. PMID- 19126034 TI - Lichen striatus in an adult following trauma with central nail plate involvement and its dermoscopy features. PMID- 19126035 TI - Treatment of parapsoriasis with bexarotene 1% gel. PMID- 19126036 TI - Lack of oncogenic mutations in the c-Met catalytic tyrosine kinase domain in acral lentiginous melanoma. PMID- 19126037 TI - Alopecia caused by the "barber ant" Pheidole pallidula. PMID- 19126038 TI - Lelis' syndrome: treatment with acitretin. PMID- 19126039 TI - Acquired unilateral nevoid telangiectasia in a 51-year-old female. PMID- 19126040 TI - Human orf complicated with erythema multiforme. PMID- 19126041 TI - Coexisting perianal squamous cell carcinoma, Bowen's disease, and condylomata acuminata treated with topical imiquimod 5%. PMID- 19126043 TI - Necrobiotic xanthogranuloma. PMID- 19126044 TI - Ornamental tattoos and skin lesions. Tattoo inoculation borderline tuberculoid leprosy. PMID- 19126045 TI - The changing patterns of hair density and thickness in South Korean women with hair loss: clinical office-based phototrichogram analysis. AB - BACKGROUND: The principal objective of this study was to investigate the clinical office-based phototrichogram (PT) findings of multiple scalp areas in South Korean women with hair loss. METHODS: Two hundred and fifty-eight women with hair loss were classified into 155 female pattern hair loss (FPHL) and 103 telogen effluvium (TE) patients. Hair density and thickness were measured via computer software-assisted PT. Eleven scalp sites, including the three points of the front, top, and back of the head and the right and left sides of the head area, were targeted in this investigation. RESULTS: PT profiles of the patients with hair loss showed significantly reduced hair density and thickness relative to the controls. The TE group showed the same order distribution as the control group, but the density and thickness were significantly lower than those of the controls. The values for the FPHL patient group declined with increasing Ludwig grade. The curve patterns of mean hair density and thickness by scalp site were shown to differ from group to group and to have a peculiar distribution. CONCLUSION: The results of this study indicate that hair density and thickness vary over the entire scalp area in patients suffering from hair loss. The measurement of multiple scalp sites and the observation of changing patterns of hair parameters are very informative in terms of differentiating less apparent hair loss disorders. Through this study, we wish to introduce a doctor- and patient-friendly hair evaluation tool for use in clinic offices. PMID- 19126046 TI - The prevalence of melasma and its association with quality of life in adult male Latino migrant workers. AB - BACKGROUND: Melasma is a common condition of Latino women that detracts from their quality of life (QOL). The prevalence and impact of melasma in Latino men is not well characterized. AIM: To assess the prevalence of melasma and its association with QOL in Latino men from Mexico and Central America working in the USA. METHODS: The prevalence of melasma was assessed in three studies of Latino men: by direct examination in a study of 25 Latino poultry workers; by direct examination in a study of 54 Latino farm workers; and by examination of store-and forward teledermatology images in a study of 300 Latino farm workers. QOL was assessed with a Spanish version of the Dermatology Life Quality Index (DLQI). RESULTS: The prevalence of melasma was 36.0%, 7.4%, and 14.0% in the three studies. The prevalence of melasma was greatest amongst those aged 31 years and older, who were from Guatemala, and who spoke an indigenous language. The presence of melasma was associated with higher DLQI scores, indicating a poorer QOL, in the poultry worker population. CONCLUSIONS: Melasma is a common condition in Latino men and is associated with a poor QOL in some affected individuals. Clinicians should be aware that melasma may be a concern for their male Latino patients. Research on the association of skin conditions with QOL amongst minority men is needed. PMID- 19126047 TI - Polymerase chain reaction compared to other laboratory findings and to clinical evaluation in the diagnosis of cutaneous tuberculosis and atypical mycobacteria skin infection. AB - BACKGROUND: Cutaneous tuberculosis has re-emerged in the last 15 years together with the higher incidence of pulmonary tuberculosis and multidrug resistance. The choice for a single diagnostic tool among the many available today is a challenge. Our objective was to compare polymerase chain reaction (PCR) with other exams in the diagnosis of cutaneous tuberculosis and atypical mycobacteria skin infection. METHODS: PCR and a set of five different exams were performed in 32 patients (34 samples of paraffin-embedded tissue) evaluated for 3 years in a university hospital, considering the response to mycobacterial infection treatment as a positive case. RESULTS: PCR was the most sensitive (88%) and specific (83%) exam. Culture, immunohistochemistry and acid-fast bacilli were not in agreement with clinical response to treatment. Conclusions Although PCR is a useful tool, careful clinical exam is still the gold standard for the evaluation and treatment of cutaneous tuberculosis and mycobacteria skin infection. PMID- 19126048 TI - Herpes simplex virus: a histopathologic study of the depth of herpetic wounds. AB - BACKGROUND: Herpes is a prevalent, infectious disease that can occur anywhere on the body; it is found primarily on the face and genitalia. Herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) are the DNA viruses that cause human herpes. Clinically, HSV-1 and HSV-2 infections produce lesions generally located on the mucocutaneous junctions of the face and genitalia. At times, vesicular lesions may ulcerate, leaving recalcitrant wounds that are challenging to treat. Until now, the basis of treatment has been related to the eradication of the viral infection. Little attention has focused on the consequence of the viral infection and the resulting wounds, specifically whether this represents an epidermal or dermal injury. METHODS: Using 10 herpetic lesions from different individuals, we studied the depth of the injury via routine hematoxylin and eosin stains, as well as periodic acid-Schiff (PAS) and type IV collagen stains, which demonstrate the presence of the basement membrane. RESULTS: In all cases, we found an inflammatory infiltrate in the dermis and selective disruption of the basement membrane. CONCLUSION: This suggests that herpetic lesions involve the dermis and are best classified as partial-thickness wounds. PMID- 19126049 TI - Comparison of depression, anxiety and life quality in acne vulgaris patients who were treated with either isotretinoin or topical agents. AB - BACKGROUND: Since the introduction of isotretinoin to the market, many adverse psychiatric effects, including depression, anxiety and suicide attempts were reported. The aim of this study was to determine whether patients with acne who were treated with isotretinoin experienced significant increases in psychiatric symptoms over a 4-month period compared with patients who received topical acne therapy. METHODS: Seventy-eight acne patients were allocated either to isotretinoin treatment (study group) (n = 37) or to topical treatment (control group) (n = 41). Their psychological status was evaluated at the baseline, second and fourth months of the treatment. All patients were required to complete the dermatology life quality index (DLQI), the Hospital anxiety and depression (HAD) scale, and the beck depression inventory (BDI). RESULTS: The two groups were not different from each other in terms of DLQI, BDI, HAD-A, HAD-D and total HAD scores at baseline. However, at the end of the second month quality of life was more impaired in the topical treatment group compared to the isotretinoin group (P < 0.05), and there were no difference between two groups in terms of BDI, HAD A, HAD-D, and total HAD scores (P > 0.05). At the end of fourth month quality of life and all psychological test scores had improved more in the isotretinoin group compared to topical treatment group (P < 0.05). CONCLUSION: Results of the present study indicate that there is no increase in depressive and anxiety symptoms in the isotretinoin treatment group compared to that in the topical group. Instead, successful treatment of acne seems to improve both depressive and anxiety symptoms and improve quality of life. PMID- 19126050 TI - A novel mutation in the ATP2C1 gene is associated with Hailey-Hailey disease in a Chinese family. AB - BACKGROUND: A three-generation Chinese family with Hailey-Hailey disease (HHD) was identified and characterized. The proband developed HHD with severe recurrent blisters and crusted erosions involving the body folds. Skin biopsy studies showed epidermal hyperkeratosis and defects in cell-to-cell adhesion. Three other members in the family were also affected with HHD and had the same clinical manifestations. The purpose of this study was to identify the pathogenic gene or mutation in the family. METHODS: All exons and exon-intron boundaries of ATP2C1 were polymerase chain reaction (PCR) amplified and sequenced with DNA samples from the proband. Restriction fragment length polymorphism (RFLP) analysis for the intron 23-exon 24 boundary of ATP2C1 was performed in all family members and in 100 normal control subjects. RESULTS: A novel 2-bp deletion (c.2251delGT) was detected in exon 24 of the ATP2C1 gene. The mutation was present in the three other affected family members and in two asymptomatic young carriers, but not in the other normal family members or the 100 normal controls. The mutation resulted in a frameshift change and led to the formation of a premature termination codon (PTC) four amino acid residues downstream from the sixth transmembrane domain. CONCLUSIONS: Our results indicate that the novel c.2251delGT (p.V751fs) mutation in the ATP2C1 gene is responsible for HHD in this Chinese family. This study expands the spectrum of ATP2C1 mutations associated with HHD. PMID- 19126051 TI - Evaluation of treatment with pentamidine for cutaneous leishmaniasis in Suriname. AB - BACKGROUND: In Suriname, pentamidine isethionate (PI) is the only drug available for the treatment of cutaneous leishmaniasis (CL). Recently, local dermatologists have observed an increase in CL patients not responding adequately to the standard doses. METHODS: In this study, patient compliance to PI treatment was assessed, and its efficacy was evaluated by comparing the clinical criteria and parasitologic load in week 3 of treatment. Skin biopsies were collected before, during and at the end of therapy and tested by quantitative nucleic acid sequence based amplification. RESULTS: In total, 67 patients with suspected CL were enrolled during the recruitment period, of which only 23 patients with confirmed CL were followed until the end of treatment. All 23 patients were found to be infected with Leishmania (Viannia) guyanensis. A lower cure rate (76-78%) was estimated than that obtained previously (90%), and only 50% of the recruited CL patients finished the complete treatment schedule. CONCLUSIONS: As one-half of the CL patients were treated insufficiently, a much shorter treatment protocol should be considered to improve the inadequate follow-up. PMID- 19126052 TI - Cutaneous leishmaniasis associated with visceral leishmaniasis in a case of acquired immunodeficiency syndrome (AIDS). AB - INTRODUCTION: Leishmania/human immunodeficiency virus (HIV) coinfection is emerging as an increasingly frequent and extremely serious new disease. Although many reports have described the association of visceral leishmaniasis and AIDS, cutaneous leishmaniasis associated with AIDS is very uncommon. CASE SUMMARY: We describe a case of visceral leishmaniasis/HIV coinfection associated with cutaneous Leishman body-positive lesions in a patient from Jahrom, a city in Fars province in Iran. CONCLUSION: Our case demonstrated that it is better to evaluate the diagnosis of visceral leishmaniasis in patients who present with cutaneous leishmaniasis and HIV infection. PMID- 19126053 TI - Primary cutaneous CD4+/CD56+ hematodermic neoplasm (blastic NK-cell lymphoma): a case report. PMID- 19126054 TI - Neutrophilic dermatosis of the hands after influenza vaccination. AB - An otherwise healthy 72-year-old man presented with a painful eruption composed of grouped hemorrhagic purulent blisters on erythematous plaques, on both palms of his hands, which appeared 12 h after he had been vaccinated against influenza. Based on the patient's history, physical examination, histopathological and laboratory findings, the diagnosis of neutrophilic dermatosis of the hands (NDH) was made. NDH is currently considered a distributional variant of Sweet's syndrome (SS). It is identical to atypical pyoderma gangrenosum (PG) and pustular vasculitis of the hands. There are only seven reported cases of SS after vaccination, none of them with lesions confined solely to the palms or soles. Our current case is the third one of SS following influenza vaccination. PMID- 19126055 TI - Autochthonous leprosy in metropolitan France presenting with a diffuse infiltration of the face and febrile illness. AB - Leprosy is now considered in Western Europe as a strictly imported disease. A case of presumed locally acquired diffuse lepromatous leprosy was observed in a native-Portuguese woman living in France who developed an acute febrile presentation with extensive cutaneous signs. PMID- 19126056 TI - An atypical presentation of erythema elevatum diutinum involving palms and soles. PMID- 19126057 TI - Late-onset Papillon-Lefevre syndrome with pyogenic liver abscesses: report of one case. PMID- 19126058 TI - Does tachyphylaxis occur in long-term management of scalp seborrheic dermatitis with pyrithione zinc-based treatments? AB - BACKGROUND: Scalp seborrheic dermatitis and dandruff (SD/D) are chronic conditions requiring long-term treatment. There is a common belief that patients frequently experience decreasing benefits over time when using a single product. This physiologic accommodation is termed tachyphylaxis. AIM: To systematically investigate the anecdotal belief that tachyphylaxis occurs in long-term treatment of SD/D using quantitative clinical assessments. METHODS: An international questionnaire completed by 722 dermatologists assessed the belief of tachyphylaxis incidence with pyrithione zinc (PTZ)-based shampoos, time course, occurrence relative to active ingredients, and effect of switching products. Two double-blind, randomized, clinical evaluations were conducted, 24- and 48-week studies, whereby a 1% PTZ shampoo, a 2% PTZ shampoo, or a matched placebo control shampoo was used by each subject for the duration of the study. Dermatologists assessed the adherent scalp flaking (scale of 0-10) at baseline and at specified intervals. RESULTS: Sixty-four per cent of responding dermatologists believed tachyphylaxis occurred with PTZ products, and most felt that tachyphylaxis occurred within 3 months of use. Evaluation of mean treatment responses vs. placebo and individual responses as a function of study duration showed a consistent benefit for all products at all time points; therefore, no evidence for tachyphylaxis was found (within 48 weeks of treatment). CONCLUSION: No evidence for tachyphylaxis in SD/D treatment by PTZ-based shampoos was found. Compliance could explain the decreasing response rate seen over time; the solution is to choose an affordable therapeutic product that is effective long term without cosmetic trade-offs. PMID- 19126059 TI - Efficacy and safety of tarcrolimus cream 0.1% in the treatment of vitiligo. AB - BACKGROUND: Vitiligo is an acquired, pigmentary skin disorder which is disfiguring and difficult to treat. Phototherapy and application of topical corticosteroids are most commonly prescribed. However, these therapies are often not effective and use of corticosteroids on the face may lead to cutaneous atrophy, telangiectasia, and ocular complications. OBJECTIVE: We sought to assess the efficacy of topical tacrolimus ointment in the treatment of vitiligo. METHODS: A prospective pilot study was performed of 30 patients with vitiligo. Patients were treated with tacrolimus ointment for at least 4 months. Clinical responses were documented during clinic visits, and by pretacrolimus and post tacrolimus photography. RESULTS: Twenty-five (83.3%) patients showed some repigmentation at the end of 4 months. Patients with vitiligo for more than 5 years also responded well to tacrolimus ointment. Repigmentation in active vitiligo was superior to that in stable vitiligo. 80% of patients with segmental vitiligo of the head and neck showed some response to tacrolimus, but there was no statistical significance between segmental and vulgaris vitiligo. The mean percentage of repigmentation on the head and neck was greater than that on the trunk and extremities. Four patients initially experienced burning on application. CONCLUSIONS: Topical tacrolimus ointment is an effective and well tolerated alternative therapy for vitiligo especially involving the head and neck. PMID- 19126060 TI - What can primary cutaneous neuroendocrine carcinomas with squamoid and neuroendocrine differentiation teach us about the origin of Merkel cells? PMID- 19126061 TI - Erythema ab igne following Sauna belt use for abdominal obesity and cellulite. PMID- 19126062 TI - Multiple microvenular hemangiomas in a healthy child. PMID- 19126063 TI - A case of trichomegaly of the eyelashes and facial hypertrichosis induced by erlotinib (Tarceva). PMID- 19126064 TI - A case with Stevens Johnson syndrome triggered by combination of clobazam, lamotrigine, and valproic acid treatment. PMID- 19126065 TI - Cushing's syndrome caused by mucosal corticosteroid therapy. PMID- 19126067 TI - Claims, contradictions and country life in Australia: the evidence on rural nursing and midwifery. PMID- 19126068 TI - The Tri-focal model of care: advancing the teaching-nursing home concept. AB - Providing residential aged care is challenging because of the complexity of residents' health status, difficulties recruiting and retaining skilled staff, and financial and regulatory constraints. This paper discusses some of these challenges and describes an innovative model of care, termed 'The Tri-focal model of care'. This model was developed based on the concepts of 'partnership-centred care', 'positive work environment' and the need for evidence-based practice to underpin all aspects of care. It is envisaged that the implementation of this model will provide a rich learning environment that advances the teaching-nursing home concept and the quality of residential aged care. PMID- 19126069 TI - Quality of life in Jordanian children with asthma. AB - The purpose of this study was to describe how Jordanian children with asthma experience their quality of life (QoL). Two hundred children with asthma (34% girls and 66% boys) participated in the study. The Paediatric Asthma Quality of Life Questionnaire was used to measure how children with asthma disease impaired their daily life during the previous week on three domains: 'activity limitations', 'symptoms' and 'emotional function'. The main finding of this study was that children with asthma scored their QoL towards the negative end of the scale, but they scored more limitations in the domain of activities than in emotions and symptoms. Children reported that the most restricted activity was their ability to run. Younger children, being a girl and living in the rural areas, were associated with lower QoL. The children in the present study might not comply with their health regimen, and this might gave a darker view of the daily life of children with asthma. Further studies of Jordanian children and their QoL were suggested to identify and support the factors which influence QoL of children with asthma and other chronic diseases in Jordan. PMID- 19126070 TI - A non-randomized comparison of e-learning and classroom delivery of basic life support with automated external defibrillator use: a pilot study. AB - This pilot study investigated whether computer-based learning package followed by practical instruction and traditional classroom methods were comparable in developing knowledge and skills in basic life support with automated external defibrillator. Eighty-three mental health care professionals were allocated to one of two groups. Twenty-eight completed an e-learning package, and the remaining 55 received delivery of content in a classroom. Using standardized assessment methods, comparisons of participant knowledge gain and performance in resuscitation were made. Significant increases in knowledge followed training. No differences were found with basic resuscitation skills or in the time taken to the first shock; however, both groups were inaccurate with electrode pad placement. E-learners performed slightly better in 21 of the 30 observed skills. Overall group performance did not differ suggesting computer-based education has the potential to prepare learners in resuscitation knowledge and skills to comparable levels of classroom courses. PMID- 19126071 TI - Awareness of risk factors for coronary heart disease following interventional cardiology procedures: a key concern for nursing practice. AB - Cardiovascular risk factor modification to prevent progression of coronary heart disease is important for patients following percutaneous coronary intervention. The aims of this study were to assess patient's awareness of modifiable cardiac risk factors and examine if patients with modifiable risk factors were more likely to identify these risk as amenable to change. Awareness of risk factors was measured using the Indiana Cardiac Rehabilitation Knowledge Questionnaire in a cohort of prospective, consecutive participants post percutaneous coronary intervention. Completed questionnaires were received from 75% of the participants. The majority were able to identify high cholesterol (87%), smoking (83%) and hypertension (82%) as modifiable risk factors. Less than half (46%) of the respondents identified diabetes as a modifiable risk factor. Only a third of participants recognized all six modifiable risk factors. A large proportion of patients who were smokers, or who had high cholesterol or hypertension, identified these as risk factors. A third of people with documented diabetes did not recognize this condition as a risk factor for heart disease. The findings have important implications for nursing practice in terms of directing educational efforts for the modification of risk factors for coronary heart disease. PMID- 19126072 TI - The impact of endometriosis on work and social participation. AB - Endometriosis is a debilitating chronic disease that can affect many aspects of everyday life owing to symptoms such as pain and fatigue. This paper reports the findings of a study exploring the impact of symptomatic endometriosis on women's social and working life. The study used a feminist approach. Eighteen women were interviewed and a thematic approach used to analyse the data. The analysis is structured around three themes focusing on issues around disclosure of symptoms in the work place; the impact of symptoms on work, education and social participation; and the strategies used by women in the study to manage endometriosis. A range of health and employment implications are discussed in this paper. In particular, nurses can provide useful support to women by careful assessment and prompt referral for diagnostic procedures and by providing timely and comprehensive information, including information about the lifestyle and nutritional factors recommended by women with endometriosis. PMID- 19126073 TI - Implementation and evaluation of critical thinking strategies to enhance critical thinking skills in Middle Eastern nurses. AB - The purpose of this study was to develop, implement and evaluate critical thinking strategies to enhance critical thinking skills in Middle Eastern nurses. Critical thinking strategies such as questioning, debate, role play and small group activity were developed and used in a professional development programme, which was trialled on a sample of Middle Eastern nurses (n = 20), to promote critical thinking skills, encourage problem solving, development of clinical judgment making and care prioritization in order to improve patient care and outcomes. Classroom learning was transformed from memorization to interaction and active participation. The intervention programme was successful in developing critical thinking skills in both the nurse educators and student nurses in this programme. This programme successfully integrated critical thinking strategies into a Middle Eastern nursing curriculum. Recommendations are as follows: (1) utilize evidence-based practice and stem questions to encourage the formulation of critical thinking questions; (2) support the needs of nurse educators for them to effectively implement teaching strategies to foster critical thinking skills; and (3) adopt creative approaches to (i) transform students into interactive participants and (ii) open students' minds and stimulate higher-level thinking and problem-solving abilities. PMID- 19126074 TI - Severity of fatigue is related to functional limitation and health in patients with chronic obstructive pulmonary disease. AB - Fatigue is one of the most prevalent symptoms in patients with chronic obstructive pulmonary disease (COPD). In research as well as in clinical practise, fatigue and its influence on functioning and health has not been in focus. The aim of this study was to compare fatigue, functional limitations owing to fatigue and health between patients with COPD and individuals from the general population to assess the differences between patients experiencing no, moderate and severe fatigue. Patients with COPD (n = 151) and individuals from the general population (n = 95) answered questions about fatigue, the Fatigue Impact Scale and the Medical Outcomes Survey Short Form-36. The patients with COPD reported a higher frequency, longer daily duration and more severity of fatigue compared with individuals from the general population as well as more functional limitations and worse health. The patients who reported severe fatigue had more functional limitations and worse health compared with patients reporting moderate fatigue. These results indicate that fatigue severity should be screened for during the nursing care process with purpose to reduce the symptom burden. PMID- 19126075 TI - Older people with delirium: worthless and childlike. AB - Delirium occurs as a consequence of physiological imbalances resulting in alterations in consciousness and cognitive impairment. It is a serious cognitive disorder and one prevalent in older people. This paper presents a significant finding from a study that critically examined the major discourses circulating on delirium and the subject positions offered to older people who had been delirious. A qualitative research design utilizing a critical gerontological framework underpinned this study. Data sources included published documents on delirium and semi-structured taped interviews. A discourse analytic approach interrogated the 20 sets of data collected. Textual analysis revealed the existence of an ageist discourse. Consequently, older people who had been delirious were offered the subject positions of 'being old doesn't matter' and 'a second childhood'. However, results from this study identified times when people who had been delirious resisted the ageist discourse. Resistance to the dominant discourse, although subtle and occurring on a micro level, acted to destabilize and challenge ageist and discriminatory health practices. PMID- 19126076 TI - Adaptation and validation of the Iranian version of Angina Pectoris characteristics questionnaire. AB - Research aimed at evaluating Angina Pectoris (AP) symptoms in patients with Coronary Heart Disease (CHD) requires a comprehensive assessment tool applicable to this population. Because no single existing measure assessed the full range of symptom characteristics important to our study, we combined two measures (the modified Rose Questionnaire (MRQ), and the Short Form-McGill Pain Questionnaire (SF-MPQ) and added one item to examine specific aspects of AP symptoms (Accompanying Symptom Checklist). The purposes of this study are: (i) to describe the process of translating the SF-MPQ, MRQ and accompanying symptom checklist from English to the Farsi language; and (ii) to evaluate substantial psychometric works (content and construct validity) for the entire instrument and each part of it (SF-MPQ, MRQ and accompanying symptom checklist). A cross-cultural adaptation of the instrument was carried out before developing the final version of it. To these ends, one hundred and 10 CHD patients with AP were assessed by this hybrid measure in an initial study. An analysis of the results indicated that a content valid instrument had been developed. There was a significantly high correlation between scores in the SF-MPQ and there were associations between items in the MRQ. The Iranian version of AP characteristics questionnaire is valid; it is capable of describing characteristics of AP symptoms. However, the reliability of the instrument needs to be assessed through further research. PMID- 19126078 TI - Near misses: paradoxical realities in everyday clinical practice. AB - This qualitative study was conducted to define and describe what constitutes and contributes to near miss occurrences in the health-care system and what is needed to ensure safer processes of care. Nine health-care organizations (13 sites total) including six academic health sciences centres (acute care, mental health and geriatric) and three community hospitals participated in this study. The final sample consisted of 37 focus groups (86 in the nursing staff only; 62 in the pharmacy staff only; and 99 in the mixed nursing and pharmacy focus groups respectively) and 120 interviews involving 144 health-care consumers. Data were collected using focus groups (health-care professionals) and key informant interviews (health-care consumers). A multi-level content analyses schema (transcription, coding, categorizing, internal consistency, thematic analysis and community validation) was used. Six themes emerged from the multi-level content analyses that combined focus group (health-care professionals) and key informant interview (health-care consumers) data. These themes are discussed under the three original research questions with supporting data derived from codes and categories. Study findings implicate changes for the health-care landscape relative to system, health policy, professional development and quality improvement. PMID- 19126077 TI - Being an interviewer in a structured oral assessment process for selection of undergraduate nursing students: motivations, expectations and reflections of the experience. AB - There is a dearth of knowledge about individuals who interview candidates for nursing. Who are they and why do they volunteer to take part in an oral assessment process for selection of undergraduate nursing students? The aim of this study was to answer these questions, and to seek the perspectives of the assessors about the process. Academic staff of the Faculty of Health Sciences, staff members of the Discipline of Nursing and registered nurses working in affiliated hospitals nominated for roles as oral assessors. All assessors received training and no payment was made. Thirty-nine of 45 (86.6%) oral assessors completed questionnaires on their experiences of being an assessor. The majority of assessors were female (69.2%) and from a clinical background (71.8% vs. 28.2% faculty-based). The most commonly cited motivations included a desire to contribute to the development of the nursing profession (94.9% of responding); because they knew what was involved in a nursing career (79.5%); wanting to influence who becomes a nurse (76.9%); and because they understood the qualities needed to be a registered nurse (76.9%). Assessors spoke highly of the experience and in general found it enjoyable. PMID- 19126079 TI - Incontinence: managed or mismanaged in hospital settings? AB - This paper reports the results of a survey of inpatients to determine the prevalence of their continence status and the overall management of their incontinence. A survey of 447 hospitalized adults was conducted and an audit of their medical records. Twenty-two per cent of patients reported urinary incontinence, 10% faecal incontinence, 78% nocturia, 23% urinary urgency and 11% trouble passing urine. Pre-existing bladder and bowel problems were reported by 34% and 26% of patients respectively. Sixty per cent of patients were using a continence product or device. There was a lack of documentation in the medical records about patients' continence status and about their pre-admission bowel and bladder status. The findings reveal that the management of incontinence in acute and subacute settings is suboptimal. There is a need to raise clinical awareness about incontinence in hospital settings and to implement a structured approach to its assessment and management. Furthermore, as the costs associated with the management or mismanagement of incontinence in hospital settings are not fully understood, there is a need for further research on this issue. PMID- 19126081 TI - Letter from the United States. PMID- 19126082 TI - 'Caesura' as Bion's discourse on method. AB - The author contends that Caesura, one of Bion's last works, can be read as the equivalent of Descartes's Discourse on Method. In this compact and complex text, the dictate of 'methodical' and 'hyperbolic doubt'- so called because it is taken to the extreme form of application to the faculty of thought itself - which, for Descartes, represents the fundamental principle of philosophical and scientific research, is reflected in the formula of 'transcending the caesura'. Bion directs his attention successively to the pairs of opposing concepts that structure psychoanalytic discourse and demonstrates their paradoxical and non-separative logic. The binary system of producing meaning is deconstructed through the systematic use of non-pathological - i.e. not static but dynamic - reversible perspective. A viewpoint that appears natural, self-evident and primary is plunged into crisis and proves to be founded on what the punctuation mark of the slash excludes. Yet the new point of view does not supplant its predecessor, but supplements it. The conceptual opposition is not overturned, but merely destabilized in such a way as to maintain a creative tension that generates new thoughts. By this technique of wrong-footing the reader, Bion achieves what is tantamount to a Kuhnian revolution: the transition from Freud's semiotic or evidential paradigm to an aesthetic one, centred on emotional experience - to a 'science of at-one-ment'. Working with the antithetical concepts of censorship and caesura, the author illustrates some clinical implications of this radical shift. PMID- 19126083 TI - Three boys and their stories: atypical eating and primitive relations. AB - Eating disorders of whatever category, especially the most severe ones such as anorexia or bulimia, are often considered typical of the female sex. While this is correct concerning these two disorders, I suggest that there are other eating disorders that are more frequent among boys, especially the ones concerning atypical relations to food, notwithstanding the evidence that the already very well-known syndromes of bulimia and anorexia are also gaining ground among males. However, similar, unspecified eating disorders do not seem to occur so often among girls. Clinical data from my private practice should be taken for granted considering these matters, since I do not yet have statistical data to back up my thesis. In examples from three clinical cases involving boys and one female vignette, I shall attempt to establish a relation between eating disorders and primitive relations with men and women. This relation goes well beyond the oft cited 'contemporary' causes of eating disorders. In addition, I shall situate these differences among symptoms of both sexes and relate them to impasses in different stages of emotional development, as well as to the extent to which girls' identification with their mother's sex can influence their acquisition of eating disorders. PMID- 19126084 TI - Biographical truths and their clinical consequences: understanding 'embodied memories' in a third psychoanalysis with a traumatized patient recovered from severe poliomyelitis. AB - The relationship between 'narrative' and 'historical-biographical truth' in psychoanalytic treatment has become the subject of many controversial debates in recent years. Findings of contemporary memory research have lead to great scepticism as to whether therapists are able objectively and reliably to reconstruct biographical events on the basis of their observations in the therapeutic situation. Some authors even claim that psychoanalysts should concentrate exclusively on observing the here and now of the patient's behaviour within the transference relationship to the analyst. In this paper it will be discussed whether the baby has been thrown out with the bathwater in this debate. Centred around the insights from a third psychoanalysis with a patient who suffered from a severe case of childhood polio, the hypothesis will be discussed that working through the traumatic experience in the transference with the analyst, as well as the reconstruction of the biographical-historical reality of the trauma suffered, prove to be indispensable for a lasting structural change. Integration of the trauma into one's own personal history and identity is and remains one of the main aims of a psychoanalytic treatment with severely traumatized patients. The reconstruction of the original trauma is indispensable in helping the patient to understand the 'language of the body' and to connect it with visualizations, images and verbalizations. The irreversable wounds and vulnerability of his body as the 'signs of his specific traumatic history' have to be recognized, emotionally accepted and understood in order to live with them and not deny them any longer. Another important aspect in psychoanalysis is to develop the capability to mentalize, in other words, to understand the intentions of central (primary) objects related to the trauma. The concept of 'embodied memory' might be helpful in understanding precisely in what way 'early trauma is remembered by the body'. Observing in detail the sensory-motor coordinations in the analytic relationship enables one to decode the inappropriate intensity of affects and fantasies which match the original traumatic interaction and are revealed as inappropriate reactions in the present, new relationship to the analyst. PMID- 19126085 TI - Donald Woods Winnicott (1896-1971): a brief introduction. AB - This essay offers a guide to the trajectory of Winnicott's theories from 1919 to 1971. Part one surveys the archives, publications, and the collected works project. Part two is divided into four sections to illustrate one foundation phase followed by three major theoretical phases. Each section outlines the evolution of Winnicott's thought and presents the seminal conceptions of each phase. At the end of each section the publications and key papers are tabulated with brief notes. Table 5 shows the posthumous publications. A new chronology is added. PMID- 19126086 TI - On: reading Harold Searles. PMID- 19126088 TI - On: Racial fantasies and the primal scene of miscegenation. PMID- 19126089 TI - On: Research in analytically oriented group psychotherapy: A tool for the evaluation of adherence to the technique in studies of effectiveness. PMID- 19126090 TI - Film essay: Saying yes to dirt, desire and difference: Yes (2004). PMID- 19126091 TI - Film essay: Michel Deville's La lectrice: Honouring or deriding Freud's theories through bibliotherapy? PMID- 19126100 TI - Abstracts of the Australasian Society of Clinical Immunology and Allergy (ASCIA) 19th Annual Scientific Meeting. November 12-14, 2008. Melbourne, Australia. PMID- 19126101 TI - Effect of a xenograft on early bone formation in extraction sockets: an experimental study in dog. AB - AIM: The aim of this study was to study the effect on early bone formation resulting from the placement of a xenograft in the fresh extraction socket in dogs. MATERIAL AND METHODS: Five beagle dogs were used. The distal roots of the third and fourth mandibular premolars were removed. In one quadrant, a graft consisting of Bio-Oss Collagen was placed in the fresh extraction wound, while the corresponding premolar sites in the contra-lateral jaw quadrant were left non grafted. After 2 weeks of healing, the dogs were perfused with a fixative, the mandibles removed, the experimental sites dissected, demineralized, sectioned in the mesio-distal plane and stained in hematoxyline-eosine. RESULTS: The central portion of the non-grafted sockets was occupied by a provisional matrix comprised of densely packed connective tissue fibers and mesenchymal cells. Apical and lateral to the provisional matrix, newly formed woven bone was found to occupy most of the sockets. In the apical part of the grafted sockets, no particles of the xenograft could be observed but newly formed bone was present in this portion of the experimental site. In addition, limited numbers of woven bone trabeculae occurred along the lateral socket walls. The central and marginal segments of the grafted sockets, however, were occupied by a non-mineralized connective tissue that enclosed Bio-Oss particles that frequently were coated by multinucleated cells. CONCLUSIONS: The placement of Bio-Oss Collagen in the fresh extraction wound obviously delayed socket healing. Thus, after 2 weeks of tissue repair, only minute amounts of newly formed bone occurred in the apical and lateral borders of the grafted sockets, while large amounts of woven bone had formed in most parts of the non-grafted sites. PMID- 19126102 TI - Evaluation of a new biodegradable membrane to prevent gingival ingrowth into mandibular bone defects in minipigs. AB - OBJECTIVE: The aim of this study was to test whether a synthetic, biodegradable membrane made of polyethylene glycol (PEG) can prevent soft-tissue ingrowth into alveolar defects. MATERIAL AND METHODS: In each of 16 minipigs, three mandibular premolars were bilaterally extracted. Three months later, acute standardized defects (diameter 8 mm, depth 8 mm) were prepared. Four treatment modalities were randomly allocated to the defects: (1) PEG membrane plus collagen sponge, (2) polylactide (PLA) membrane plus collagen sponge, (3) collagen sponge alone, and (4) empty defect. Animals were sacrificed at 10 days (n=5), 21 days (n=5), or 2 months (n=6) after treatment. Qualitative and quantitative histological evaluations of soft-tissue ingrowth and bone regeneration were performed on nondecalcified ground sections. For statistical analysis, the Mann-Whitney Wilcoxon test, the Kruskal-Wallis, and the paired t-test were applied. P-values were adjusted using the Dunnett-Hsu adjustment. RESULTS: At 10 days, the PEG membrane group showed the least soft-tissue ingrowth (mean value -0.75 mm; range 1.35 to -0.10), followed by the PLA membrane group -0.18 mm (-0.80 to 0.44), the collagen group 0.04 mm (-0.65 to 0.73), and the empty defects 0.60 mm (-0.08 to 1.29). Statistically significant differences were observed between the PEG membrane group and the empty defects (P<0.05). At 21 days, the highest percentage of newly formed bone was found in the PEG membrane group (mean 28.4%; range 21.6 35.2) compared with 23.7% (16.9-30.5; PLA membrane), 15.2% (8.2-22.2; collagen group), and 21.6% (14.5-28.8; empty defects). Statistically significant differences were only found between the PEG membrane group and the collagen group (P<0.05). At 2 months, the tested parameters revealed no statistically significant differences between the groups. CONCLUSION: The experimental PEG membrane applied in the present study successfully prevented collapse of the covering soft tissues to a degree similar to the PLA membrane. The combination of a collagen sponge and the PEG membrane showed the least soft-tissue ingrowth at 10 days and promoted more bone formation at 21 days. PMID- 19126103 TI - Human osteoclast formation and activity on an equine spongy bone substitute. AB - OBJECTIVES: The aim of the present study was to evaluate the in vitro formation and activity of human osteoclasts (OCLs) generated on a new type of xenograft for bone substitution, an equine spongy bone. MATERIAL AND METHODS: Peripheral blood mononuclear cells from healthy volunteers were used to generate OCLs in vitro in the presence of macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL) on bovine bone slices (positive control) and equine spongy bone. Morphological and biochemical methods were used to assess OCLs formation and activity. RESULTS: Cells generated after 21 days of culture on equine spongy bone showed similar morphology to those on the positive control and displayed typical OCL markers and features, indicating that this material supported OCL formation. Moreover, these cells were functionally active on equine spongy bone with statistically significant differences compared with the control in the release of tartrate-resistant acid phosphatase (TRAcP5b) at days 14 and 21 of culture. With regard to the resorption, on equine bone, OCLs formed smaller discontinuous island-like lacunae rather than the typical lobulated, tracking resorption lacunae observed on the control. CONCLUSIONS: This study enables clinicians to tailor the usage of equine spongy bone and presents a model, which can be applied to the preclinical assessment of bone substitute material's resorbability and resorption rates. PMID- 19126104 TI - Ectopic bone formation after implantation of a slow release system of polylactic acid and rhBMP-2. AB - OBJECTIVES: The present study was conducted to test the hypothesis that preshaped polylactic acid (PLA) implants loaded with recombinant human bone morphogenic protein 2 (rhBMP-2) can induce bone formation in a rat ectopic model. MATERIALS AND METHODS: Two groups of porous cylindrical poly-DL-lactic acid implants of 8 mm diameter were produced by gas foaming with CO(2), incorporating 48 and 96 microg rhBMP-2, respectively, into each implant. Blank PLA implants were used as controls. The release of BMPs and the induction of alkaline phosphatase were assessed in vitro. Osteoinduction in vivo was tested by insertion of 15 implants from each group into the gluteal muscles of Wistar rats. Five implants from each group were retrieved after 6, 13 and 26 weeks and assessed using flat panel volume detector computed tomography and light microscopy. RESULTS: Both groups of implants showed increased release of rhBMP-2 during the first 24-48 h, with a slightly higher amount being released from the implants with 48 microg. Release during subsequent intervals was <100 ng/72 h in the low-concentration group and >100 ng in the group with 96 microg rhBMP-2. Implants with 95 microg rhBMP-2 exhibited bone formation in vivo on the outside of the implants across the observation period of 26 weeks with invasion of bone into the pores, whereas implants with 48 microg rhBMP-2 failed to induce the formation of bone tissue. No bone formation was found in the control implants. CONCLUSIONS: The results suggest that release rates of rhBMP-2 for ectopic bone induction have to be >100 ng/72 h to maintain the osteoinductive activity of the tested porous PLA implants. This slow release system may have impact on alveolar bone augmentation procedures when used as individually preformed osteoinductive implants. PMID- 19126105 TI - Effect of modifications of dual acid-etched implant surfaces on peri-implant bone formation. Part I: organic coatings. AB - OBJECTIVE: The aim of the present study was to test the hypothesis that peri implant bone formation can be improved by modifying dual acid-etched (DAE) implant surfaces using organic coatings that enhance cell adhesion and osteogenic differentiation. MATERIAL AND METHODS: Ten adult female foxhounds received experimental titanium implants in the mandible 3 months after removal of all premolar teeth. Six types of implants were evaluated in each animal: (i) implants with a machined surface (MS), (ii) implants with a DAE surface topography, (iii) implants with an acid-etched surface coated with RGD peptides, (iv) implants with an acid-etched surface coated with collagen I, (v) implants with an acid-etched surface coated with collagen I and chondroitin sulphate (CS), (vi) implants with an acid-etched surface coated with collagen I and CS and recombinant human bone morphogenetic protein-2. Peri-implant bone regeneration was assessed by histomorphometry after 1 and 3 months in five dogs each by measuring bone implant contact (BIC) and the bone volume density (BVD) of the newly formed peri-implant bone. RESULTS: After 1 month, mean BIC was significantly higher in the coated implants group than in the MS group. There was no significant difference when mean BIC in the DAE group was compared with implants with any of the organic coatings, but the difference was significant when compared with the MS implants. Differences in mean BVD value did not reach significance between any of the surfaces. After 3 months, the same held true for the mean BIC of all the groups except for Coll I. Mean volume density of the newly formed bone was higher in all the surface modifications, albeit without statistical significance. CONCLUSIONS: It is concluded that with the exception of Coll I, the tested organic surface coatings on DAE surfaces did not improve peri-implant bone formation when compared with the DAE surfaces but enhanced BIC when compared with the MSs. PMID- 19126106 TI - Effect of modifications of dual acid-etched implant surfaces on periimplant bone formation. Part II: calcium phosphate coatings. AB - The aim of the present study was to test the hypothesis that calcium phosphate coatings of dual acid-etched surfaces (DAEs) can improve periimplant bone regeneration. Ten adult female foxhounds received experimental titanium screw implants in the mandible 3 months after removal of all premolar teeth. Five types of surface states were evaluated in each animal: (i) implants with a machined surface (MS) (Control 1); (ii) implants with a DAE (Control 2); (iii) implants with a DAE coated with collagen I (Control 3); (iv) implants with a DAE with mineralized collagen I; and (v) implants with a DAE with a hydroxylapatite (HA) coating. Periimplant bone regeneration was assessed by histomorphometry after 1 and 3 months in five dogs each by measuring bone implant contact (BIC) and the volume density of the newly formed periimplant bone (BVD). After 1 month, mean BIC of experimental implants did not differ significantly from implants with DAE and collagen-coated surfaces, but was significantly higher than the MS implants. BVD was enhanced significantly only in implants with mineralized collagen coating compared with DAE and collagen-coated controls. After 3 months, the mean values of BIC had increased significantly in the group of implants with HA and mineralized collagen coating but were not significantly different from implants with DAE and collagen-coated surfaces. The same held true for the mean BVD values. In conclusion, the present study could not verify the hypothesis that calcium phosphate coatings of DAEs in the present form enhanced periimplant bone formation compared with the DAE surface alone. PMID- 19126107 TI - Bone quality in the midpalate for temporary anchorage devices. AB - OBJECTIVE: The aim of the present study was to quantitatively assess the bone quality of the palatal bone from an implantologic standpoint. MATERIAL AND METHODS: The material consisted of palatal tissue blocks of autopsy material from 22 subjects (19 male, three female), between 18 and 63 years of age. The specimens comprised the anterior part of median palate (APMP) from about 7 mm behind the incisive foramen (first premolar regions), the middle part of median palate (MPMP, second premolar region), and the posterior part of the median palate (PPMP, first molar region). They were prepared in the transversal plane using ground-thin-section technology. The midpalatal areas of the different parts of the palate [regions of interest (ROI) for temporary anchorage device (TAD) placement] 3 mm bilaterally to the midline were assessed with respect to hard tissue fraction to total bone volume (HTF/TBV), bone marrow fraction to total bone volume (BMF/TBV), and sutural tissue fraction to total bone volume (STF/TBV). Furthermore, age-specific evaluation was performed: younger age group (18-25 years, n=10) and older age group (26-63 years, n=12). RESULTS: Histomorphometric measurement showed the following results for the APMP: mean HTF/TBV, 68.57%; mean BMF/TBV, 21.38%; and mean STF/TBV, 9.96%. There were no statistically significant differences between the various tissue fractions of the APMP, MPMP, and PPMP. The younger age group showed a mean HTF/TBV of 68.88%, a mean BMF/TBV of 21.5%, and a mean STF/TBV of 9.53%. No statistically significant differences with the older age group were found. CONCLUSION: These results suggest that the HTF in the APMP, MPMP, and PPMP in younger as well as older adults is relatively high. Thus, good primary stability of TADs should be achieved in all parts of the adult median palate independent of age. PMID- 19126108 TI - Behavior of two osteoblast-like cell lines cultured on machined or rough titanium surfaces. AB - BACKGROUND: Two osteosarcoma-derived cell lines have been extensively used to investigate the biological events occurring on titanium surfaces: MG63 and Saos 2. However, the behavior of the two lines on different titanium surfaces has never been compared. AIM: The aim of the present study was to compare the behavior of MG63 and Saos-2 cells on two different titanium surfaces, machined and rough (sandblasting and acid-etched). We compared cell proliferation and morphology, alkaline phosphatase (ALP) activity and secretion of osteocalcin (OC). RESULTS: The most pronounced difference between the two cell lines was that ALP activity in the Saos-2 cells was 10-fold higher than in the MG63 cells. The proliferation rate of the MG63 cells was much higher than that of the Saos-2 cells at all the tested cell concentrations. MG-63 cells, but not Saos-2 cells, grown on rough surface titanium proliferated more rapidly than cells grown on machined surfaces. Morphological analysis revealed that Saos-2 cells and cells grown on the rougher surface, displayed a more mature phenotype. The level of OC secreted by the Saos-2 cells, but not the MG63 cells, were higher on the rough surface than on the machined surface. CONCLUSIONS: This study shows that Saos-2 cells exhibit a more mature osteoblast phenotype, compared with that of MG63 cells, rendering them a good candidate for an in vitro model of osseointegration. PMID- 19126109 TI - Pore characteristics of bone substitute materials assessed by microcomputed tomography. AB - OBJECTIVES: Pore configurations of alloplastic biomaterial scaffolds play a major role for new bone formation in vivo. Current studies on characteristics of pores in bone substitute materials focus on individual particles or single blocks. Thus, three-dimensional (3-D) architecture of particle aggregates, representing the clinical relevant in vivo situation is not adequately taken into account. The aim of this study was the visualization and quantification of pore properties, both of the scaffold structure of single particles as well as of the micro morphology of complex 3-D aggregated particle-conglomerates. MATERIAL AND METHODS: In model experiments, standardized plexiglass cylinders were stuffed with commercial bone substitute material particles with diverse chemical composition (HA, beta-TCP, HA-SiO(2), HA-beta-TCP, bioactive glass), origin (phycogenic, bovine, synthetic) and granulation (50 mum-2000 mum). Analogue to establish procedures for native (human) bone samples, non-fixed bone substitute materials were scanned by high-resolution microcomputed tomography. In addition to computer animated two-dimensional and 3-D reconstruction of the samples, median pore thickness and pore size distribution were determined. Materials representative for their chemical constitution were documented by SEM imaging. RESULTS: Investigated specimens significantly were different in micro-morophology and pore properties, ranging from highly porous to rather solid. The most voluminous pores were localized interparticularly. Within one product line, the determined pore properties showed a significant correlation with single particle grain sizes. CONCLUSION: The generation and interpretation of micro-CT based 3-D pore models can provide further insight into the expected osteoconduction dynamics and therefore might serve as a basis for further modifications of scaffold size and geometry as well as for further invasive studies on the biological behaviour of the scaffolds. PMID- 19126110 TI - Bleeding on probing and pocket probing depth in relation to probing pressure and mucosal health around oral implants. AB - OBJECTIVES: To assess the bleeding on probing (BOP) tendency and periodontal probe penetration when applying various probing forces at implant sites in patients with a high standard of oral hygiene with well-maintained peri-implant tissues. MATERIAL AND METHODS: Seventeen healthy patients with excellent oral hygiene in a maintenance program after having been treated for periodontitis or gingivitis were recruited. Missing teeth had been replaced using oral implants. The BOP and probing depth (PPD) were assessed at the mid-buccal, mid-oral, mesial and distal aspects of the buccal surfaces of each implant. Moreover, contralateral teeth were designated and assessed for BOP and PPD in the same locations and at the same observation visits. At each visit, implants and contralateral teeth were randomly assigned to one of the standardized probing forces (0.15 or 0.25 N). The second probing force was applied at the repetition of the examination 7 days later. RESULTS: Increasing the probing pressure by 0.1 N from 0.15 N resulted in an increase of BOP percentage by 13.7% and 6.6% for implants and contralateral teeth, respectively. There appeared to be a significant difference of the mean BOP percentage at implant and tooth sites when a probing pressure of 0.25 N was applied. A significantly deeper mean PPD at implant sites compared with tooth sites was found irrespective of the probing pressure applied. CONCLUSIONS: The results of the present study demonstrated that 0.15 N might represent the threshold pressure to be applied to avoid false positive BOP readings around oral implants. Hence, probing around implants demonstrated a higher sensitivity compared with probing around teeth. PMID- 19126111 TI - Marginal fit of cemented and screw-retained crowns incorporated on the Straumann (ITI) Dental Implant System: an in vitro study. AB - AIM: The aim of this study was to assess the marginal fit of crowns on the Straumann (ITI) Dental Implant System with special consideration of different casting dental materials. MATERIAL AND METHODS: Sixty porcelain-fused-to-metal crowns were fabricated: 18 crowns on standard cone abutments with an impression cylinder, partially prefabricated analogs, no coping and screw-retained (A); 18 crowns on solid abutments without an impression device, no analogs, no coping and cemented (B); and 18 crowns on solid abutments using an impression transfer cap, an analog with a shoulder, no coping and cemented (C). In each group, six crowns were made on epoxy mastercasts (Bluestar), six on synthetic plaster (Moldasynt) and six on super hard stone (Fujirock). Six additional crowns were fabricated with the transversal screw retention system onto the Octa system with impression transfer caps, metal analogs, gold copings and screw-retained (D). Impregum was used as impression material. Crowns of B and C were cemented with KetacCem. Crowns of A and D were fixed with an occlusal screw torqued at 15 N cm. Crowns were embedded, cut and polished. Under a light microscope using a magnification of x 100, the distance between the crown margin (CM) and the shoulder (marginal gap, MG) and the distance between the CM and the end of the shoulder (crown length, CL) was measured. RESULTS: MGs were 15.4+/-13.2 microm (A), 21.2+/-23.1 microm (B), 11+/-12.1 microm (C) and 10.4+/-9.3 microm (D). No statistically significantly differences using either of the casting materials were observed. CLs were -21.3+/-24.8 microm (A), 3+/-28.9 microm (B), 0.5+/-22 microm (C) and 0.1+/-15.8 microm (D). Crowns were shorter on synthetic casting materials compared with stone casts (P<0.005). CONCLUSIONS: CMs fit precisely with both cemented and screw-retained versions as well as when using no, partial or full analogs. PMID- 19126112 TI - CAD/CAM fabrication and clinical application of surgical template and bone model in oral implant surgery. AB - OBJECTIVES: A novel implant surgery support system with computer simulation for implant insertion and fabrication of a surgical template that helps in drilling bone was developed. A virtual reality haptic device that gives the sense of touch was used for simulation and a surgical template was fabricated by CAD/CAM method. Surgical guides were applied for two clinical cases. MATERIAL AND METHODS: Three dimensional (3D) jaw bone images transferred from DICOM data filmed by CT scanner were fed to the software and manipulated using the haptic device. The site for implant insertion was determined after evaluating the quality of bone and position of the mandibular canal. The surgical template was designed with ease using the free design CAD function of haptic device. The surgical template and bone model were fabricated by a fused deposit modeling machine. Two clinical cases were applied using the present system. RESULTS: Simulation to determine the site of implant insertion and fabrication of the surgical bone templates were successfully done in two clinical cases, one for three implant insertion in lower right jaw and the other is for seven implant insertion in lower edentulous jaw, respectively. During surgery, the templates could be firmly adapted on the bone and drilling was successfully performed in both cases. CONCLUSION: The present simulation and drilling support using the surgical template may help to perform safe and accurate implant surgery. PMID- 19126113 TI - Placement of Branemark implants in the maxillary tuber region: anatomical considerations, surgical technique and long-term results. AB - BACKGROUND: Fixture placement in the tuber area is one way to overcome the problem of insufficient bone volume for routine implant surgery in the posterior maxilla due to severe resorption of jawbone and an extensive enlargement of the maxillary sinus. However, little is known about the long-term results. PURPOSE: The aim of this study was to retrospectively evaluate the survival rate and marginal bone conditions at fixtures placed in the tuber region of the maxilla. MATERIAL AND METHODS: Twenty-one patients previously treated with at least one implant in the tuber region of the maxilla were included in this retrospective analysis. A total of 23 standard Branemark System fixtures with a turned surface had been surgically placed in the tuber regions and 71 additional implants in adjacent areas to support fixed dental bridges. All implants were allowed to heal for 6-8 months before abutment connection and following prosthetic treatment. The patients were radiographed after 1-12 years for evaluation of marginal bone levels. In addition, the relation between the apex of the fixture in the tuberosity area and the posterior border of the maxilla was measured. RESULTS: Twenty of the 21 patients representing 22 tuber and 64 additional implants were radiographically evaluated. No implants in the tuber areas were lost during the follow-up whereas two fixtures in the anterior region had to be removed, one before loading and the other after 4 years of loading not interfering with the prosthesis stability. The mean marginal bone level at tuber implants was situated on average 1.6 mm (SD 1.1, n=22) from the abutment-fixture junction, whilst the other implants showed an average bone level of 1.9 mm (SD 0.8, n=64). The results were similar when comparing partially and totally edentulous patients. CONCLUSION: The present retrospective study shows good clinical outcome with standard Branemark fixtures placed in the tuber region of the posterior maxilla using a two-stage procedure. In appropriate cases where bone of adequate volume and density is available, our data indicate that the technique can be used as an alternative to more extensive surgery and especially to the sinus lift procedure. However, prospective comparative studies are needed in order to evaluate the efficacy of the described technique for this purpose. PMID- 19126114 TI - Short-term clinical and microbiological evaluations of peri-implant diseases before and after mechanical anti-infective therapies. AB - OBJECTIVES: The aim of this study was to evaluate the clinical and microbiological effects of mechanical anti-infective therapies for mucositis and peri-implantitis. MATERIAL AND METHODS: Subjects with at least one dental implant were assigned to healthy (n=10), mucositis (n=12) or peri-implantitis (n=13) groups. Implants with mucositis or peri-implantitis were decontaminated by means of teflon curettes and abrasive sodium carbonate air-powder, performed by an open flap for peri-implantitis and without surgery for mucositis. Visible plaque (PI), marginal bleeding (MB), bleeding on probing (BOP), suppuration (SUP), probing depth (PD) and relative clinical attachment level (rCAL) were assessed at baseline and at 3 months after therapies. At the same time points, submucosal plaque samples were collected from each implant and analyzed by Checkerboard DNA DNA hybridization for 40 bacterial species. RESULTS: All clinical parameters improved at 3 months post-therapy in mucositis and peri-implantitis groups (P<0.05). The mean reduction in rCAL (+/-SD) was 1.4+/-1.2 mm and 2.3+/-1.6 mm, and it was 1.3+/-1.2 mm and 3.1+/-1.7 mm in PD (+/-SD) for mucositis and peri implantitis, respectively. Levels of Treponema denticola, Tanerella forsythia and Parvimonas micra, and of Fusobacterium nucleatum ss nucleatum, were significantly reduced after peri-implantitis therapy and after mucositis therapy, respectively (P<0.05). In addition, counts of Porphyromons gingivalis, Treponema socranskii and the proportions of red complex were reduced in both groups at 3 months after treatments (P<0.05). CONCLUSION: Mechanical therapies alone were effective in treating mucositis and peri-implantitis over a period of 3 months. The open debridement procedure showed clinical and microbiological benefits on the treatment of peri-implantitis and could be safely used as a standard control group for future studies. PMID- 19126115 TI - Dedicated chairs in headache science: the way forward? PMID- 19126116 TI - New daily persistent headache in the paediatric population. AB - We conducted a clinic-based study focusing on the clinical features of new-onset chronic daily headaches (CDH) in children and adolescents. The clinical records and headache diaries of 306 children and adolescents were reviewed, to identify 187 with CDH. Relevant information was transferred to a standardized form that included operational criteria for the diagnoses of the headaches. Since we were interested in describing the clinical features of these headaches, we followed the criteria A and B of the 2nd edn of the International Classification of Headache Disorders (ICHD-2) and refer to them as new daily persistent headaches (NDPH) regardless of the presence of migraine features (therefore, this is a modified version of the ICHD-2 criteria). From the 56 adolescents with NDPH, most (91.8%) did not overuse medications. Nearly half (48.1%) reported they could recall the month when their headaches started. NDPH was more common than chronic tension-type headache in both adolescents overusing and not overusing medication. Individuals with NDPH had headaches fulfilling criteria for migraine on an average of 18.5 days per month. On most days, they had migraine-associated symptoms (one of nausea, photophobia or phonophobia)). NDPH is common in children and adolescents with CDH. Most subjects do not overuse medication. Migraine features are common. PMID- 19126117 TI - Effect of adrenomedullin on the cerebral circulation: relevance to primary headache disorders. AB - Adrenomedullin (ADM) is closely related to calcitonin gene-related peptide, which has a known causative role in migraine. Animal studies have strongly suggested that ADM has a vasodilatory effect within the cerebral circulation. For these reasons, ADM is also likely to be involved in migraine. However, the hypothetical migraine-inducing property and effect on human cerebral circulation of ADM have not previously been investigated. Human ADM (0.08 microg kg(-1) min(-1)) or placebo (saline 0.9%) was administered as a 20-min intravenous infusion to 12 patients suffering from migraine without aura in a crossover double-blind study. The occurrence of headache and associated symptoms were registered regularly 24 h post infusion. Cerebral blood flow (CBF) was measured by (133)Xenon single-photon emission computed tomography, mean blood flow velocity in the middle cerebral artery (V(MCA)) by transcranial Doppler and the diameter of peripheral arteries by transdermal ultrasound (C-scan). ADM did not induce significantly more headache or migraine compared with placebo (P = 0.58). CBF was unaffected by ADM infusion (global CBF, P = 0.32 and rCBF(MCA), P = 0.38) and the same applied for the V(MCA) (P = 0.18). The superficial temporal artery dilated compared with placebo (P < 0.001), and facial flushing was seen after ADM administration (P = 0.001). In conclusion, intravenous ADM is not a mediator of migraine headache and does not dilate intracranial arteries. PMID- 19126118 TI - Parental psychopathology and migraine headaches among adolescent girls. AB - Migraine headaches and depression often co-occur within individuals, and both syndromes run in families. However, knowledge about how these disorders relate across generations, as well as how migraine relates to other forms of psychopathology, is sparse. This study examined risk for migraine among female adolescent offspring of parents with different types of psychopathology. The sample was drawn from the Minnesota Twin Family Study, a community-based study of adolescents and their families (n = 674, 17-year-old female adolescents and their biological parents). Diagnoses of maternal, paternal and offspring major depression, antisocial behaviour, alcohol dependence and drug dependence were based on structured interviews. Migraine headaches in each family member were assessed via interviews with the mother. Parental depression, antisocial behaviour and drug dependence were associated with offspring migraine. These associations mostly remained significant even when parental migraine and the corresponding type of psychopathology in offspring were adjusted for. In contrast, there were no significant associations between parental psychopathology and offspring stomach problems, indicating that these associations did not extend to all offspring somatic symptoms. These results emphasize the need to look at antisocial behaviour and substance-related problems when examining associations between migraine and psychopathology, and indicate that more research on inter generational links between migraine and psychopathology is needed. PMID- 19126119 TI - Excitatory and inhibitory purinergic control of neck muscle nociception in anaesthetized mice. AB - Tension-type headache is associated with noxious input from neck muscles. Due to the importance of purinergic mechanisms in muscle nociception, experimental studies typically inject alpha,beta-methyleneadenosine 5'-triphosphate (alpha,beta-meATP). In contrast to native adenosine 5'-triphosphate (ATP), alpha,beta-meATP has a narrow receptor profile and remains stable in tissue. The present study administered alpha,beta-meATP or ATP in semi-spinal neck muscles in anaesthetized mice (n = 65) in order to address different effects in neck muscle nociception. The jaw-opening reflex monitored the impact of neck muscle noxious input on brainstem processing. Injection of alpha,beta-meATP induced reflex facilitation in a dose-dependent manner. In contrast, only the lowest ATP dosage evoked facilitation. Preceding P2Y(1) receptor blockade revealed facilitation even under high-dosage ATP. Ongoing facilitation after alpha,beta-meATP injection neutralized under subsequent activation of P2Y(1) receptors. Results demonstrate opposing excitatory P2X and inhibitory P2Y effects of ATP in neck muscle nociception. These mechanisms may be involved in the pathophysiology of neck muscle pain in man. PMID- 19126126 TI - The systematic review in otolaryngology: a guide to preparation. PMID- 19126128 TI - Treatment of non-tuberculous mycobacterial cervicofacial lymphadenitis in children: critical appraisal of the literature. AB - BACKGROUND: Surgical excision has historically been the treatment of choice for non-tuberculous mycobacterial cervicofacial lymphadenitis. Emerging data suggests antibiotic treatment alone could be an attractive alternative to surgery. We questioned (1) what treatment offers best cure rates in children with this condition and (2) the evidence for antibiotic treatment alone. TYPE OF REVIEW: Structured literature search according to the Evidence Based Medicine guidelines. SEARCH STRATEGY/METHODS: A structured search was conducted in PubMed, Embase and the Cochrane Library from 1966 up to November 2007. Relevant papers were critically appraised. RESULTS: Six papers were included, one of which was a randomised controlled clinical trial (RCT). In general, treatment sequencing was not consistent. Surgery and medical treatment were often used in combination, resulting in high overall cure rates. In the RCT surgery and antibiotic treatment were compared as single modalities. The outcome of surgery was far superior (96%versus 66% cure rate, respectively). Other studies indicated that substantial cure rates (up to 67%) were achieved with medical management alone, but failed to identify factors predicting response. CONCLUSION: Surgical excision still is the backbone in the management of cervicofacial non-tuberculous mycobacterial lymphadenitis, although a considerable number of children can be cured with antibiotics alone or combined modality treatment. To date it is unclear which subset of patients can benefit from antibiotic treatment only. PMID- 19126129 TI - Prospective cohort comparison of bioactive glass implants and conchal cartilage in reconstruction of the posterior canal wall during tympanomastoidectomy. AB - OBJECTIVE: To compare the effectiveness of bioactive glass implants and conchal cartilage in reconstructing the posterior canal wall during tympanomastoidectomy. STUDY DESIGN: Prospective cohort clinical study. SETTING: Teaching hospital. PATIENTS: Patients with clinically diagnosed chronic suppurative otitis media and cholesteatoma awaiting tympanomastoidectomy were recruited. INTERVENTION: All patients underwent tympanomastoidectomy by the same surgeon. A first cohort of 12 patients underwent posterior canal wall reconstruction with autogenous conchal cartilage. A second cohort of 12 patients underwent such reconstruction with prefabricated bioactive glass. MAIN OUTCOME MEASURES: Primary- All patients underwent out-patient review at 1, 3, 6 and 12 months postoperatively, after which a second-look procedure was performed. Reconstructions were inspected for evidence of epithelialization, granulation, infection, stenosis, depression and extrusion. Secondary- All patients had perioperative serial pure-tone audiometry to check for any change in hearing levels upto 1 year postoperatively. RESULTS: By 1 year postoperatively, both reconstructive graft materials showed good epithelialization, no granulation, no infection, no ear canal stenosis, no depression and no extrusion. At operative second-looks, bioactive glass particularly showed good tissue bonding, including both neovascularization and connective tissue integration. Overall clinical outcome was equivalent for both materials. Both graft materials showed no statistically significant difference in postoperative hearing levels. CONCLUSIONS: Bioactive glass and conchal cartilage showed equivalent clinical outcome in reconstructing the posterior canal wall without significantly affecting hearing levels. As bioactive glass does not require second site morbidity and thus also reduces operative time, we prefer it for reconstructing the posterior canal wall following tympanomastoidectomy. PMID- 19126127 TI - Combined electro-acoustic stimulation: a beneficial union? AB - BACKGROUND: The most pressing problem facing cochlear implant research is no longer making artificial hearing a reality. Instead, it is to develop devices that can more clearly reflect the capabilities of the human auditory system. Current cochlear implants rarely provide adequate pitch perception. As hearing loss commonly affects higher, more than lower frequencies, a possible solution is to preserve acoustic hearing at low frequencies by inserting a short electrode array and thus deliver combined electro-acoustic stimulation (EAS). OBJECTIVE OF REVIEW: To determine whether individuals with severe-to-profound high-frequency hearing loss have realised this predicted benefit of combined EAS, over conventional cochlear implants, with respect to pitch. TYPE OF REVIEW: A systematic review of publications pertaining to the benefits of combined EAS over conventional cochlear implantation, with specific reference to pitch perception. SEARCH STRATEGY: A systematic literature search was conducted across multiple databases and supplemented by searching the reference lists of relevant trials and identified reviews. RESULTS: The included studies suggest an overall benefit of combined EAS, over conventional cochlear implants, with respect to pitch. In addition, (i) 13% sustained a total loss of low-frequency hearing post implantation of the short electrode array and, (ii) 24% had >20 dB hearing loss across all frequencies and/or total hearing loss. CONCLUSIONS: For patients with severe-to-profound high-frequency hearing loss combined EAS appears to offer a significant, everyday, long-term benefit. However, further clinical trials with larger numbers of candidates are necessary to confirm this finding. The risks involved cannot be ignored, but there is potential for a variety of strategies to improve the safety margin. PMID- 19126130 TI - Speech perception in congenitally, pre-lingually and post-lingually deaf children expressed in an equivalent hearing loss value. AB - OBJECTIVES: To investigate the speech perception performance of children with a cochlear implant (CI) after 3 and 4 years of follow-up and to study the influence of age at implantation, duration of deafness and communication mode on the variability in speech perception performance. STUDY DESIGN: A broad battery of speech perception tests was administered to 67 children with a CI. The results were reduced into one measure: the 'equivalent hearing loss (EHL)'. This outcome measure refers to the performance of a reference group of severely and profoundly hearing impaired children with conventional hearing aids. PARTICIPANTS: The population comprised 35 congenitally, 17 pre-lingually and 15 post-lingually deaf children implanted between 1989 and 1999. The population was homogeneous with respect to cognition, residual hearing and support at home as a result of conservative inclusion criteria. RESULTS: During the first 2 years after implantation, post-lingually deaf children showed the fastest rate of improvement in speech perception. After 3 years of implant use, the early implanted pre lingually deaf children and congenitally deaf children implanted under the age of 6 years caught up with the post-lingually deaf children. Pre-lingually deaf children implanted after a relatively long-duration of deafness tended to show poorer performance than those with a shorter duration. Performance of congenitally deaf children implanted after the age of 6 years was poorer and progress was slower. In the congenitally deaf children, 36% of the variability in performance was explained by duration of deafness, whereas in the children with pre- and post-lingually acquired deafness, communication mode explained 69% of the variance. CONCLUSIONS: All children derived benefit from their CI for speech perception tasks, but performance varied greatly. Several children reached EHL levels around 70 dB; their speech perception was equal to that of a child with conventional hearing aids who has 70 dB HL. After early implantation, the levels of performance that were eventually achieved differed no more than 10 dB, irrespective of whether the onset of deafness was pre-lingual or postlingual. In congenitally deaf children, duration of deafness played a major role in speech perception performance, whereas in the children with pre-lingually and post lingually acquired deafness together, mode of communication was a major factor. PMID- 19126131 TI - The effect of hand dominance on recurrent idiopathic paediatric epistaxis. AB - OBJECTIVES: No published data exists to support the belief that nose picking contributes to recurrent idiopathic paediatric epistaxis. If nose picking is implicated then there may be a higher incidence of epistaxis on the side of the dominant hand. The objective of the study was to determine whether nose picking has a significant role in paediatric epistaxis by comparing hand dominance and side of epistaxis. DESIGN AND SETTING: A prospective, consecutive cohort of new epistaxis referrals to a tertiary paediatric centre over a 14-month period were assessed for hand dominance and reported side of the epistaxis. The presence of septal excoriation, crusting and prominent blood vessels was also recorded. PARTICIPANTS: One hundred and forty-eight new epistaxis referrals. MAIN OUTCOME MEASURES: Dominant hand, side of epistaxis history and side of positive examination findings. RESULTS: No statistically significant association between hand dominance and either epistaxis history or examination findings was identified. A statistically significant association between epistaxis history and examination findings was identified P < 0.001, K = 0.614, SE 0.571. CONCLUSIONS: The study does not provide evidence to support the belief that paediatric epistaxis is influenced by nose picking. The study provides evidence that patients' epistaxis history and positive examination findings are a consistent marker of previous epistaxis. PMID- 19126132 TI - Sensitivity and responsiveness of the Medical Research Council dyspnoea scale to the presence and treatment of adult laryngotracheal stenosis. AB - OBJECTIVES: To assess the sensitivity and responsiveness of the Medical Research Council (MRC) scale, a psychophysical dyspnoea assessment instrument to the presence and treatment of adult laryngotracheal stenosis. DESIGN: Prospective observational study. SETTINGS: Tertiary/National referral airway reconstruction centre. PARTICIPANTS: Fourty tracheostomy-free patients undergoing endoscopic airway examination/laryngotracheoplasty. MAIN OUTCOME MEASURES: Demographic and clinical information, obtained from patient records, lesion severity, which was recorded intraoperatively, standard spirometry, which was measured preoperatively, and the MRC dyspnoea scale, which was administered preoperatively and at the first outpatient visit 4-6 weeks later. RESULTS: There were 16 males and 24 females. Mean age at presentation was 44 +/- 14 years (+/- SD). Postintubation stenosis was the commonest aetiology (73%) followed by idiopathic subglottic stenosis and Wegener's Granulomatosis. Six patients were examined post treatment and had minimal residual stenosis and the remaining patients had glottic stenosis (n = 11) or Myer-Cotton Grade I (n = 8), II (n = 7) or III (n = 8) tracheal stenoses. Pre-treatment MRC dyspnoea scores and the degree of change in the MRC score following treatment strongly correlated with pre-treatment stenosis severity (r = 0.75 and r = -0.71 respectively; P < .001). Moreover statistically significant correlations existed between preoperative peak expiratory flow and forced expiratory volume in 1 s and preoperative MRC dyspnoea scores (r = -0.34 and r = -0.35 respectively; P < 0.05). DISCUSSION: Exertional dyspnoea is the hallmark symptom of laryngotracheal stenosis and for many patients it is the primary cause of disability. These findings confirm that the MRC dyspnoea scale is an appropriate outcome instrument for assessing dyspnoea associated with this condition. PMID- 19126133 TI - The inflammation produced by corticosteroid inhalers in the pharynx in asthmatics. AB - OBJECTIVES: To investigate inflammation of the pharyngeal mucosa caused by inhaled corticosteroids. DESIGN: Prospective, cross-sectional, single-blinded study. SETTING: University Hospital Aintree, Liverpool, UK. PATIENTS: Fifty adults were recruited from two local general practices and from general ENT clinics at our University hospital. Patients were allocated to one of four groups according to use of inhaled corticosteroids and the presence of adverse local side effects. MAIN OUTCOME MEASURES: Scores achieved on a respiratory symptom questionnaire. Histological markers of inflammation and their correlation with pharyngitis. Statistical modelling included univariate and multivariate analyses, which included multiple linear and logistic regression, and discriminant analysis. RESULTS: The regular use of inhaled corticosteroids predisposed subjects to hoarseness, weakness of voice, sore throat and throat irritation (P < 0.0001). Pharyngitis was significantly different between the groups (P < 0.0001). Furthermore, those not using an inhaled corticosteroid regularly had little or no clinically apparent pharyngitis, whereas those using an inhaled corticosteroid regularly had significantly higher pharyngitis scores (P = 0.0204). Similarly, weakness of voice (P = 0.0234), hoarseness (P < 0.001) and sore throat (P < 0.001) were also more common in those patients that used an inhaled corticosteroid on a regular basis. To our surprise, however, cellular markers of inflammation did not corroborate the appearances of clinical examination. We found that the five most important discriminators, between those that were using inhaled corticosteroid therapy regularly and those that were not, to be intra epithelial inflammatory cells (scdf -1.2939); age (scdf 0.8389); use of a spacer device (scdf 0.5456); sore throat (scdf 0.4230) and throat irritation (scdf 0.4015). The groups were significantly different (P < 0.0001). The statistical model used, classified 68% of the cases correctly into their respective groups. CONCLUSIONS: Inhaled corticosteroids predispose to pharyngitis and an inflammatory infiltrate. However, the clinical diagnosis of pharyngitis does not correlate well with cellular inflammatory infiltrate and is therefore, not a reliable measure of underlying inflammation. We advocate caution in the use of pharyngeal erythema as a measure of underlying inflammation. PMID- 19126134 TI - Hemithyroidectomy for benign thyroid disease: who needs follow-up for hypothyroidism? AB - OBJECTIVE: To identify the incidence and possible risk factors for the development of hypothyroidism following hemithyroidectomy and to determine a simple protocol for the post-op monitoring of thyroid function. DESIGN: Prospective study. SETTING: Tertiary Referral Centre. PATIENTS: Eighty-two consecutive patients who underwent hemi-thyroidectomy between May 2003 and May 2005 were included. The clinical and pathological factors were obtained and statistically analysed. RESULTS: Eighteen per cent of patients became hypothyroid post-operatively. Multivariate analysis demonstrated that risk factors for hypothyroidism following hemithyroidectomy included a pre-operative [TSH] >1.6 microIU/L [P = 0.008, hazard ratio (HR) 14, 95% confidence interval (CI) 2-54] and lymphocytic infiltration graded 3+ or more (P < 0.001, HR 42, 95% CI, 8 195%). There was no relationship between age, sex or weight of resected tissue and the risk of hypothyroidism. CONCLUSION: Patients with raised pre-operative [TSH] or with lymphocytic infiltration of their thyroid specimen should undergo regular testing of thyroid function within the first year of surgery. In all other patients, thyroid function at 12 months is sufficient. PMID- 19126135 TI - Objective evaluation of tongue base snoring after the use of an oral appliance: a prospective case series. AB - Oral appliances have become increasingly popular for treatment of obstructive sleep apnoea especially for patients who are not able to tolerate continuous positive airway pressure devices. For simple snoring, oral appliances have become one of the treatments of choice despite a relative lack of scientific evidence of their efficiency. OBJECTIVES: This study was designed to objectively evaluate the clinical effectiveness of oral appliance in the treatment of simple snoring. DESIGN: Prospective case series. PARTICIPANTS: Fifteen patients with confirmed simple tongue base snoring had pre- and post-oral appliance objective assessment of their snoring loudness and duration at home. MAIN OUTCOME MEASURES: The Snore Index was calculated as the number of snores per hour slept. Patients bed partners were asked to rate the snoring severity on a Spouse Dissatisfaction Scale. RESULTS: Overall there was no significant difference in the Snore Index in the pre- and post-oral appliance recordings. Subjectively, there was a statistically significant decrease in the Spouse Dissatisfaction Scale following the use of oral appliance. CONCLUSION: Tongue base snorers had no significant reduction in their snoring with the oral appliances. There is a subjective benefit which may be due to the placebo effect. PMID- 19126136 TI - Driving standards in tracheostomy care: a preliminary communication of the St Mary's ENT-led multi disciplinary team approach. AB - OBJECTIVES: To assess tracheostomy care and improve standards following the introduction of an ENT-led multidisciplinary tracheostomy ward round service. DESIGN: Prospective third cycle audit. SETTING: Tertiary academic London hospital serving an inner city population of multi-ethnic background (St Mary's Hospital, Paddington, London). PARTICIPANTS: Patients with a tracheostomy discharged from ITU to general wards. IMPLEMENTED ACTIONS: Establishment of an ENT-led Tracheostomy Multidisciplinary Team (TMDT). Weekly TMDT ward round to manage patients with a tracheostomy. ENT-led educational and training sessions for allied healthcare professionals. MAIN OUTCOME MEASURES: Compliance with local tracheostomy care guidelines (St Mary's tracheostomy care bundle) and time to tracheostomy tube decannulation. RESULTS: Preliminary results of 10 patients show improved compliance with tracheostomy care guidelines, established in 2004, rising to 94%. Average time to decannulation was significantly reduced from 21 to 5 days (P-value = 0.0005, Mann Whitney Wilcoxon Test). The mean total tracheostomy time was reduced from 34 to 24 days although this was not statistically significant (P-value = 0.13, Mann Whitney Wilcoxon Test). CONCLUSIONS: The introduction of regular ENT-led multidisciplinary input for patients with a tracheostomy significantly improved compliance with nursing care standards. There was also a reduction in the total length of time tracheostomy tubes remain in situ, with time to decannulation significantly reduced. PMID- 19126137 TI - Surgical management of otitis media with effusion in children: summary of NICE guidance. PMID- 19126138 TI - A step further than the NICE surgical otitis media with effusion guidelines; time to consider ventilation tubes being a once only operation. PMID- 19126139 TI - Partial labyrinthectomy in the treatment of labyrinthine fistula: how we do it. PMID- 19126141 TI - Aesthetic augmentation rhinoplasty with Permacol: how we do it. PMID- 19126140 TI - Hyaluronic acid fat graft myringoplasty: how we do it. PMID- 19126142 TI - Guidelines for the management of idiopathic epistaxis in adults: how we do it. PMID- 19126143 TI - Radiofrequency thyroarytenoid myothermy for treatment of adductor spasmodic dysphonia: how we do it. PMID- 19126144 TI - Fused CT and angiography image guided surgery for endoscopic skull base procedures: how we do it. PMID- 19126146 TI - Re: Avoiding a post-nasal pack in adenoidal haemorrhage. PMID- 19126147 TI - Response to Sharp on 'Failed voice restoration: closure of the tracheo oesophageal fistula'. PMID- 19126148 TI - Re: Yorick's skull model for tonsillectomy tie training. PMID- 19126149 TI - Re: Chronic cough. PMID- 19126151 TI - Re: Hairy canals and hair removal cream. PMID- 19126150 TI - Re: Digital tonsil dissection. PMID- 19126152 TI - A teaching mirror to enhance training in ENT procedures. PMID- 19126153 TI - Temporary cover for mini-trephines. PMID- 19126154 TI - The ear trainer model. PMID- 19126155 TI - The Jain Vent Tube: a new grommet. PMID- 19126156 TI - Development of a software for objective assessment of facial symmetry. PMID- 19126157 TI - Correct site marking in otolaryngology: a prospective three-cycle audit. PMID- 19126158 TI - Endoscopic-guided retrograde technique for secondary tracheo-oesophageal puncture and valve insertion. PMID- 19126175 TI - Critical appraisal of the literature on economic evaluations of substitution of skills between professionals: a systematic literature review. AB - OBJECTIVE: Substitution of skills has been introduced to increase health service efficiency, but little evidence is available about its cost-effectiveness. This systematic review aims to identify economic evaluations of substitution between professionals, to assess the quality of the study methods applied and to value the results for decision making. METHODS: Publications between January 1996 and November 2006 were searched in Medline, Cochrane, Cinahl, database of Health Technology Assessments, EPOC and Embase. Randomized controlled trials (RCTs), cost-benefit analysis, interrupted time series design and systematic reviews were selected. The methodological quality of the papers was reviewed, using the critical appraisal of Drummond and the EPOC list. RESULTS: Eleven studies were finally included of 7605 studies: three cost-effectiveness studies, three cost minimization studies and five studies related to partial economic evaluations. Small numbers of participating professionals and several limitations in the cost valuation and the measurement of costs were identified. CONCLUSIONS: Several potential limitations influence the validity and generalizability. Full economic evaluations per se are of limited value for making decisions about substitution of skills. The tenuous relationship between structural, process and outcome variables is not sufficient investigated. For meaningfully placing the costs and consequences of substitution of skills in the context of health care and generating relevant data for decision making, it is strongly recommended to combine an economic evaluation (RCT) with an observational longitudinal study. PMID- 19126176 TI - Patients bringing information to primary care consultations: a cross-sectional (questionnaire) study of doctors' and nurses' views of its impact. AB - OBJECTIVES: To explore the perceived frequency and impact of patients bringing health information from the Internet to primary care consultations. METHOD: A questionnaire was sent to all general practitioners (GPs) and practice nurses (nurses) in a primary care trust in the North-west of England. RESULTS: The response was 52.3%, more nurses responding than GPs (61.2% vs 46.8%). A substantial majority (93.9% and 78.0%) had experienced patients bringing such information in the last 6 months, which often lengthened the consultations. Significantly more nurses than GPs felt that the information helped the consultation (87.1% vs 37.7%), but also reported that patients bringing such information was off-putting (21.9% vs 6.3%). CONCLUSIONS: Both GPs and nurses experienced patients bringing health information to consultations and significantly differed in their views about whether it helped or hindered. Primary care staff should anticipate patients' use of the Internet for health information and should actively manage patients introducing it into the consultation. PMID- 19126177 TI - Antibiotics for nasopharyngitis are associated with a lower risk of office-based physician visit for acute otitis media within 14 days for 3- to 6-year-old children. AB - OBJECTIVES: This study was designed to analyse factors potentially influencing children's return visits to physicians for symptoms of acute otitis media (AOM) within 14 days after being diagnosed with nasopharyngitis (NP), and the impact of recent antibiotic use. DESIGN: A controlled population-based pharmaco epidemiological trial in 3- to 6-year-old children conducted from January to May 2000. SETTING: Three different geographical regions in France. PARTICIPANTS: Among 2507 eligible children, 2456 could be analysed and 505 children had 634 office-based physician visits (OBPV) for NP symptoms. INTERVENTIONS: The statistical associations between antibiotics prescribed for NP and an OBPV for AOM within 14 days in a population-based study were analysed along with risk factors of AOM. MAIN OUTCOMES MEASURE: Clinical events and antibiotic use. RESULTS: During the 2 weeks following physician-diagnosed NP, antibiotic use, especially a beta-lactam, significantly decreased the risk of OBPV for AOM in children (odds ratio=0.2; 95% confidence interval=0.09-0.7; P=0.002). CONCLUSION: Antibiotics prescribed to children for NP seem to protect during the following 2 weeks against the risk of OBPV for AOM. It remains to be determined whether a subgroup at high risk of developing AOM after a viral infection exists and what might be the best strategy to adopt for NP in a national programme of optimal antibiotic use. PMID- 19126178 TI - The effectiveness of nurse-led telemonitoring of asthma: results of a randomized controlled trial. AB - RATIONALE, AIMS AND OBJECTIVES: The aim of the study was to evaluate the effects on, and the relationship between, asthma symptoms, asthma-specific quality of life and medical consumption of a nurse-led telemonitoring intervention compared with regular care in asthma in the Netherlands. METHODS: One hundred and nine asthmatic outpatients (56 children; 53 adults) were randomly assigned to the treatment arms (12-month follow-up). The control group received regular outpatient care, while the intervention group used an asthma monitor with modem at home with an asthma nurse as the main caregiver. Clinical asthma symptoms and medical consumption were measured by using diaries. Asthma-specific quality of life was measured by the (Paediatric) Asthma Quality of Life Questionnaire. RESULTS: The study population generally represented mild to moderate asthmatics. The results show improvement in follow-up, but no statistically significant difference between the groups was observed. Moderate to high correlations were found within the outcome parameters, but the most remarkable was the low and statistically significant correlation between asthma-specific quality of life (daily functioning) and the self-reported beta-2 agonists. CONCLUSION: Overall, the telemonitoring programme evaluated in this study did not significantly decrease asthma symptoms or medical consumption, or improve asthma-specific quality of life. The results showed that a telemonitoring programme on its own is not a guarantee of success. The patients' perception of asthma-specific quality of life (daily functioning) should be a key element in asthma telemonitoring programmes. PMID- 19126179 TI - Assessment of individual vaccine status in a vaccinology experts' group. AB - RATIONALE: Worldwide, experts in vaccinology have promoted the broad annual coverage of health care workers with the influenza vaccine. Furthermore, pertussis vaccination is now recommended for young adults and health care workers working with newborns. AIM: To analyse the compliance with these guidelines among experts responsible for the development or dissemination of national immunization schedules. METHOD: A cross-sectional survey was conducted in a vaccinology workshop group of French experts, using a self-administered questionnaire. RESULTS: Among 44 experts, the average rate of influenza vaccination was 69.5% (95% confidence interval, 61.6% to 77.3%) between the 2003/04 and 2005/06 flu seasons, whereas the rate of pertussis vaccination during this period was only 30%. The main reasons that the experts gave for not being vaccinated were a lack of time or simply not remembering to do so. CONCLUSION: Experts had low coverage rates for influenza and pertussis vaccination. To improve these rates, a multifaceted intervention combining audit and feedback strategy with a vaccine day is planned. PMID- 19126180 TI - Acute ophthalmic referrals from primary care--an audit and recommendations. PMID- 19126181 TI - Inhibition of endothelial cell adhesion molecule expression improves colonic hyperalgaesia. AB - Leucocyte-endothelial cell interactions are prerequisite to leucocyte infiltration and intestinal inflammation. GI270384X is a novel inhibitor of ICAM 1 and E-selectin expression and inhibits leucocyte adhesion and improves experimental colitis. We hypothesized that GI270384X maybe effective in treatment of visceral hyperalgaesia. Visceromotor behavioural responses to colorectal distension (CRD) were obtained in naive rats or rats treated with zymosan (3 h) or 2,4,6-trinitrobenzene sulphonic acid (TNBS) (4 and 30 days) or rats exposed to acute restraint stress. Studies were also performed in a high-anxiety genetic model of colonic hyperalgaesia using Wistar-Kyoto (WKY) rats. Rats were treated orally with GI270384X or vehicle either prior to or after the administration of sensitizing stimulus. The visceromotor response to CRD was significantly enhanced in all models. GI270384X attenuated the enhanced responses to distension induced by inflammatory stimuli (TNBS and zymosan) and in the high-anxiety WKY rats; however, the drug did not inhibit the hypersensitivity induced by acute restraint stress. GI270384X was most potent in the models of acute inflammatory hyperalgaesia with a minimum efficacious dose (MED) of 0.3 and 1 mg kg(-1) observed in the TNBS and zymosan models respectively. The compound was less potent in the chronic and postinflammatory models with an MED of 10 and 30 mg kg( 1) observed in the WKY and 30-day TNBS models respectively. These findings show for the first time that inhibition of leucocyte-endothelial cell interactions can have a beneficial effect on visceral hyperalgaesia associated with inflammatory and chronic anxiety states, but is less effective against stress-associated visceral hyperalgaesia. PMID- 19126182 TI - Experimental oesophagitis in the rat is associated with decreased voluntary movement. AB - Growing interest has arisen regarding the mechanism of dyspeptic symptom generation. However, no evaluation system of these symptoms in animals has been developed. In this study, we examined whether voluntary movement of rats could be a measure to assess visceral symptoms of reflux oesophagitis. A chronic acid reflux oesophagitis model was made using rats, and the size of erosions was measured. Omeprazole was administered to the oesophagitis rats for 10 days. The amount of voluntary movement was measured by an infrared sensor. Intracellular spaces in oesophageal epithelium were also measured using a emission electron microscope. NP-40 soluble and insoluble fractions of claudins were examined by Western blot. Voluntary movement was significantly lower in the oesophagitis model rats than in the sham-operated rats (P < 0.01). Although omeprazole reduced the size of erosions, it did not significantly affect the total amount of voluntary movement (r = -0.033, P = 0.916). Intracellular spaces were significantly dilated in the oesophagitis model rats and claudin-3 showed a significantly lower relative quantity in the NP-40 insoluble fraction. Omeprazole significantly increased voluntary movement of oesophagitis model rats and the relative quantity of claudin-3 in the insoluble fraction (P < 0.05). Dilated intercellular spaces and the lower level of claudin-3 may relate to the voluntary movement of oesophagitis model rats. Decreases in voluntary movement of oesophagitis model rats may reflect visceral symptoms and be able to serve as an index of chronic abdominal symptoms. PMID- 19126183 TI - Distribution of serotonin receptors and interacting proteins in the human sigmoid colon. AB - This study aimed to examine the distribution of 5-HT receptors in the human colon. 5-HT induces desensitization of the circular muscle and as this is facilitated by G-protein coupled receptor kinases (GRKs) and other proteins, we also examined their distribution. Human sigmoid colon samples were dissected into three separate layers (mucosa, taeniae coli and intertaenial strips) and RNA was amplified by RT-PCR. The 5-HT(2B) receptor and all 5-HT(7) receptor splice variants were expressed in all tissues. 5-HT(4) a,b,c and n splice variants were also expressed in all tissues and 5-HT(4d), 5-HT(4g) and 5-HT(4i) were only detected in some samples. The 5-HT(2A) receptor was seen predominantly in the intertaenial strips of the colon. Only one transcript of the serotonin transporter (SERT) was detected in the muscle layers. Variation was seen in GRK expression with GRK2 and 3 predominantly expressed in the mucosa, while GRK5 and 6 were found more commonly in the taeniae coli. PDZ (named after postsynaptic density protein, Drosophila disc large tumour suppressor and tight junction protein ZO-1) domain containing proteins, which may be involved in 5-HT receptor trafficking, were also detected throughout the sigmoid colon. The 5-HT(3A) subunit was expressed in all tissues, whereas the 5-HT(3E) subunit was mainly found in the mucosa layer while the 5-HT(3B) subunit was more common in the muscle layers. Receptor interacting chaperone (RIC-3), which is involved in transporting 5-HT(3) receptor subunits, is expressed less in mucosa compared to muscle layers. In conclusion, these results show that there is variation in distribution of 5-HT receptors and interacting proteins within the sigmoid colon that may contribute to colonic function. PMID- 19126184 TI - Immune dysfunction in patients with functional gastrointestinal disorders. AB - There is increasing evidence for involvement of the immune system in functional gastrointestinal disorder (FGID), including onset after acute gastrointestinal infections, genotypes resulting in altered cytokine expression and abnormal presence of immune cells. Our aim was to assess cellular and humoral immune responses in (i) FGIDs, compared to healthy subjects and (ii) acute vs unspecified onset FGIDs. Lymphocytic [interleukin (IL)-5, IL-10, IL-13 and interferon gamma (IFN-gamma)] and monocytic [IL-10, IL-12, tumour necrosis factor (TNF)-alpha] cytokine production was characterized at baseline and after stimulation with phytohemagglutinine and anti-CD28 or lipopolysaccharide (LPS) in controls (n = 32), irritable bowel syndrome (IBS) (n = 30), functional dyspepsia (FD) (n = 23) and non-cardiac chest pain (NCCP) (n = 15). Serum IL-6 and IL-10 concentrations were compared, and the immunophenotype was assessed using fluorescent-activated cell sorter. Findings were compared for acute vs unspecified onset FGID. Compared to controls, stimulated lymphocyte expression of IL-5 and IL-13 was enhanced in IBS, FD and NCCP (all P < 0.05). Conversely, the stimulated monocytic IL-12 and lymphocytic IL-10 expression were reduced in IBS and FD, while IFN-gamma expression was also reduced in FD patients. Except for an increase in the numbers of CD3(+)CD45RA(+)CD45RO(+) cells, no distinct cellular profile was detected. Patients with a presumed acute onset of their symptoms had higher serum IL-10 levels and more CD3(+)CD45RA(+)CD45RO(+) cells, while TNF alpha levels following stimulation with LPS were higher in FD patients reporting an acute onset. A shift towards a Th2 cytokine profile is present in FGID, while the cellular immunophenotype remains largely unchanged. Further research is indicated and could provide new therapeutic strategies for these disorders. PMID- 19126186 TI - Effect of repeated, long term capsaicin ingestion on intestinal chemo- and mechanosensation in healthy volunteers. AB - Repeated ingestion of capsaicin over a prolonged period reduces symptoms in functional dyspepsia, but initially induces upper abdominal symptoms. Sensitizing chemonociception might be the cause for this initial effect of capsaicin. The aim was to evaluate the effect of prolonged capsaicin ingestion on duodenal chemo- and mechanonociception. Healthy subjects ingested capsules containing either 0.25 mg capsaicin tid (n = 8) or placebo (n = 8) for 28 days. Before (day 0) and after (day 29) capsule ingestion the duodenum was distended with a balloon and perfused with a capsaicin solution. Mechanically and chemically induced sensation was evaluated by a graded questionnaire. Aggregate perception scores were calculated. Perception scores during balloon distensions with 12 and 18 mmHg were significantly lower after 4 weeks capsaicin when compared to baseline (P < 0.05). Balloon volumes to induce first sensation (63 +/- 14 mL (day 0) vs 92 +/- 22 mL (day 29); P < 0.05) and discomfort (101 +/- 12 mL vs 137 +/- 22 mL; P = 0.05) where significantly higher after 4 weeks capsaicin application; balloon pressures to induce sensations were not significantly different. Intraluminal capsaicin application induced first sensation after 3.4 +/- 1.5 min (day 0) and 7.5 +/- 4.6 min (day 29) (P < 0.05) and discomfort after 15.9 +/- 9.8 min and 22.4 +/- 7.3 min (P < 0.05). The quality of perception was not altered by repeated capsaicin ingestion. In the placebo group, mechano- and chemonociception remained unaltered at day 29. Four weeks ingestion of capsaicin desensitized both chemonociceptive and mechanonociceptive pathways in healthy volunteers. Symptom reduction after prolonged treatment with capsaicin in dyspeptic patients might be attributed to a dual desensitizing effect of capsaicin on chemonociceptors and mechanonociceptors. PMID- 19126185 TI - Gastric dysreflexia after acute experimental spinal cord injury in rats. AB - Gastric reflexes are mediated mainly by vago-vagal reflex circuits in the caudal medulla. Despite the fact that brainstem vago-vagal circuitry remains intact after spinal cord injury (SCI), patients with SCI at the cervical level most often present gastric stasis with an increased risk of reflux and aspiration of gastric contents. Using a miniature strain gauge sutured to the gastric surface; we tested gastric motility and reflexive gastric relaxation following oesophageal distension (oesophageal-gastric relaxation reflex) in animals 3 days after a severe spinal contusion at either the third or ninth thoracic spinal segment (acute T3- or T9 SCI, respectively). Both basal gastric motility and the oesophageal-gastric relaxation reflex were significantly diminished in animals with T3 SCI. Conversely, both basal gastric motility and the oesophageal-gastric relaxation reflex were not significantly reduced in T9 SCI animals compared to controls. The reduced gastric motility and oesophageal-gastric reflex in T3 SCI rats was not ameliorated by celiac sympathectomy. Our results show that gastric stasis following acute SCI is independent of altered spinal sympathetic input to the stomach caudal to the lesion. Our data suggest that SCI may alter the sensitivity of vagal reflex function, perhaps by interrupting ascending spinosolitary input to brainstem vagal nuclei. PMID- 19126187 TI - Does magnetic stimulation of sacral nerve roots modify colonic motility? Results of a randomized double-blind sham-controlled study. AB - Although sacral nerve root stimulation (SNS) can result in a symptomatic improvement of faecal incontinence, the mechanism of action remains unknown. The aim of this study was to assess whether short-term magnetic SNS can inhibit pharmacologically induced propulsive colonic contractions. Twelve healthy volunteers (median age: 43.5 years old) were studied on two separate occasions and randomized into either active (15 Hz, 100% output intensity for 5 s min(-1) for 30 min) or sham rapid rate lumbosacral magnetic stimulation (rLSMS). Colorectal motility was recorded with a manometric catheter located at the most proximal transducer in the left colon and the most distal, in the rectum. Colonic contractions were provoked by instilling Bisacodyl. The effects of rLSMS on colonic, sigmoid and rectal contractions were monitored and recorded after Bisacodyl instillation. The appearance of high-amplitude contractions propagated or not (HAC/HAPC) provoked by Bisacodyl instillation was significantly delayed during active compared to sham stimulation (P = 0.03). There was no difference in the characteristics of HAC/HAPC (i.e. frequency, amplitude, duration, velocity of propagation) or the motility index with active or sham stimulation. The perception of urgency tended to be decreased with rLSMS following Bisacodyl instillation. The catheter was expulsed within a median of 16.5 min (range 8-39) after Bisacodyl administration during active stimulation compared to 14 min (range 5-40) during sham stimulation (P = 0.03). This study suggests that rLSMS could delay the appearance of the first Bisacodyl-induced colonic contractions. PMID- 19126188 TI - The incretin hormones GIP and GLP-1 in diabetic rats: effects on insulin secretion and small bowel motility. AB - Incretin hormones often display inhibitory actions on gut motility. The aim of this study was to investigate if altered responsiveness to glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1) as regards insulin release and small bowel motility could bring further clarity to the pathophysiology of diabetes in the Goto-Kakizaki (GK) rat. The isolated perfused pancreas was studied in male GK and Wistar rats (controls) under euglycemic and hyperglycemic conditions. Glucose-dependent insulinotropic peptide (10 nmol L( 1)) or GLP-1 (10 nmol L(-1)) were added to the medium and perfusate was collected and analysed for insulin. Moreover, GK and Wistar rats were supplied with bipolar electrodes in the small bowel and myoelectric activity was recorded during intravenous administration of GIP (1-400 pmol kg(-1) min(-1)) or GLP-1 (0.1-20 pmol kg(-1) min(-1)). Finally, tissue was collected from GK and Wistar rats for RNA extraction. Under euglycemia, GIP and GLP-1 stimulated the initial insulin response by 10-fold in GK rats (P < 0.05). At later hyperglycemia, the insulin response to GIP and GLP-1 was blunted to about one-third compared with controls (P < 0.05). In the bowel GLP-1 was about 2.6-16.7 times more potent than GIP in abolishing the migrating myoelectric complex in the GK and control rats. Polymerase chain reaction (PCR) showed GIP and GLP-1 receptor gene expression in pancreatic islets and in small bowel. The initially high, but later low insulin responsiveness to stimulation with GIP and GLP-1 along with inhibition of small bowel motility in the GK rat indicates a preserved incretin response on motility in diabetes type 2. PMID- 19126190 TI - Neuroma of a double gallbladder: a case report. AB - We report a case of 55 year old male patient who presented with recurrent upper abdominal pain following a laparoscopic cholecystectomy. A subsequent diagnostic laparoscopy revealed the presence of a second gallbladder which was initially missed. The peculiarity of his symptoms can in part be explained by the presence of a traumatic neuroma in his second gallbladder. A subsequent cholecystectomy led to a complete resolution of this patient's signs and symptoms. As far as we know this is the first report in the literature of a traumatic neuroma in a second gallbladder. PMID- 19126189 TI - Mining for single nucleotide polymorphisms in pig genome sequence data. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) are ideal genetic markers due to their high abundance and the highly automated way in which SNPs are detected and SNP assays are performed. The number of SNPs identified in the pig thus far is still limited. RESULTS: A total of 4.8 million whole genome shotgun sequences obtained from the NCBI trace-repository with center name "SDJVP", and project name "Sino-Danish Pig Genome Project" were analysed for the presence of SNPs. Available BAC and BAC-end sequences and their naming and mapping information, all obtained from SangerInstitute FTP site, served as a rough assembly of a reference genome. In 1.2 Gb of pig genome sequence, we identified 98,151 SNPs in which one of the sequences in the alignment represented the polymorphism and 6,374 SNPs in which two sequences represent an identical polymorphism. To benchmark the SNP identification method, 163 SNPs, in which the polymorphism was represented twice in the sequence alignment, were selected and tested on a panel of three purebred boar lines and wild boar. Of these 163 in silico identified SNPs, 134 were shown to be polymorphic in our animal panel. CONCLUSION: This SNP identification method, which mines for SNPs in publicly available porcine shotgun sequences repositories, provides thousands of high quality SNPs. Benchmarking in an animal panel showed that more than 80% of the predicted SNPs represented true genetic variation. PMID- 19126191 TI - Death following bilateral complete Achilles tendon rupture in a patient on fluoroquinolone therapy: a case report. AB - INTRODUCTION: Risk of tendon rupture, especially of the Achilles tendon, is one of the many potential side-effects of fluoroquinolone therapy. Achilles tendon rupture may be painful, debilitating or, as seen in our patient, devastating. While fluoroquinolone-induced tendon rupture typically accompanies other comorbidities (for example renal impairment) or concurrent steroid therapy, our case represents a medical 'first' in that there were no such comorbidities and no steroid therapy. Furthermore, our case is remarkable in that tendon rupture was bilateral, complete, and resulted in a devastating outcome. CASE PRESENTATION: A healthy 91-year-old Caucasian man was placed on fluoroquinolone (levofloxacin) therapy for a presumed bacterial pneumonitis. Subsequently, he developed bilateral heel pain, edema, and ecchymoses leading to a diagnosis of bilateral complete Achilles tendon rupture. This drug's side-effect was directly responsible for his subsequent physical and psychologic decline and unfortunate death. CONCLUSION: Fluoroquinolones are a powerful and potent tool in the fight against bacterial infection. As a class, they are employed by primary care physicians as well as by subspecialty physicians in all areas of medical practice. However, as this case illustrates, the use of these drugs is not without risk. Attention must be paid to potential side-effects when prescribing any medication, and close follow-up with patients is a medical necessity to evaluate for these adverse reactions, especially with fluoroquinolones. PMID- 19126192 TI - Nonconstrictive epicarditis mimicking a cardiac mass in a 71-year-old Caucasian man: a case report and review of the literature. AB - INTRODUCTION: Isolated cases of epicarditis are rare. Thus far, all have occurred with constrictive physiology as most cases involve both parietal and visceral pericardium. We report the first case of asymptomatic epicarditis that involved only the visceral pericardium presenting without constrictive physiology. CASE PRESENTATION: A 71-year-old male with a history of atrial fibrillation, coronary artery disease, pericardial effusion, type-2 diabetes and hypothyroidism presented with 5 weeks of fatigue and 1 day of dizziness. Physical examination was significant for pallor and tachycardia. Laboratory analysis revealed a hemoglobin count of 7.2 g/dl and iron deficiency anemia. The patient was transfused and evaluated by endoscopic ultrasound. A polypoid mass in the gastric cardia was found and later diagnosed as gastric adenocarcinoma (staged as T1N0M0). The pericardial effusion was evaluated with transthoracic echocardiography which showed a 2.0 x 2.7 cm mass associated with the right atrium. Transesophageal echocardiography confirmed the mass but did not reveal constrictive physiology. Whole-body contrast computed tomography failed to demonstrate metastatic disease. Biopsy of the cardiac mass revealed epicarditis without parietal pericardium involvement. Partial gastrectomy was performed to remove the gastric adenocarcinoma. CONCLUSION: This is the first reported case of asymptomatic epicarditis. Our case was especially unusual because the epicarditis presented as an incidental cardiac mass. The clinical picture was complicated due to the concomitant presence of gastric adenocarcinoma and chronic pericardial effusion. This case demonstrates that epicarditis should be considered in the differential diagnosis of cardiac masses. PMID- 19126193 TI - The Multiple Sclerosis Risk Sharing Scheme Monitoring Study--early results and lessons for the future. AB - BACKGROUND: Risk sharing schemes represent an innovative and important approach to the problems of rationing and achieving cost-effectiveness in high cost or controversial health interventions. This study aimed to assess the feasibility of risk sharing schemes, looking at long term clinical outcomes, to determine the price at which high cost treatments would be acceptable to the NHS. METHODS: This case study of the first NHS risk sharing scheme, a long term prospective cohort study of beta interferon and glatiramer acetate in multiple sclerosis (MS) patients in 71 specialist MS centres in UK NHS hospitals, recruited adults with relapsing forms of MS, meeting Association of British Neurologists (ABN) criteria for disease modifying therapy. Outcome measures were: success of recruitment and follow up over the first three years, analysis of baseline and initial follow up data and the prospect of estimating the long term cost-effectiveness of these treatments. RESULTS: Centres consented 5560 patients. Of the 4240 patients who had been in the study for a least one year, annual review data were available for 3730 (88.0%). Of the patients who had been in the study for at least two years and three years, subsequent annual review data were available for 2055 (78.5%) and 265 (71.8%) patients respectively. Baseline characteristics and a small but statistically significant progression of disease were similar to those reported in previous pivotal studies. CONCLUSION: Successful recruitment, follow up and early data analysis suggest that risk sharing schemes should be able to deliver their objectives. However, important issues of analysis, and political and commercial conflicts of interest still need to be addressed. PMID- 19126194 TI - Phylogenetic studies reveal existence of multiple lineages of a single genotype of DENV-1 (genotype III) in India during 1956-2007. AB - BACKGROUND: Dengue virus type 1 (DENV-1) have been mostly circulating silently with dominant serotypes DENV-2 and DENV-3 in India. However recent times have marked an increase in DENV-1 circulation in yearly outbreaks. Many studies have not been carried out on this virus type, leaving a lacunae pertaining to the circulating genotypes, since its earliest report in India. In the present study, we sequenced CprM gene junction of 13 DENV-1 isolated from Delhi and Gwalior (North India) between 2001-2007 and one 1956 Vellore isolate as reference. For comparison, we retrieved 11 other Indian and 70 global reference sequences from NCBI database, making sure that Indian and global isolates from all decades are available for comparative analysis. RESULTS: The region was found to be AT rich with no insertion or deletion. Majority of the nucleotide substitutions were silent, except 3 non-conservative amino acid changes (I --> T, A --> T and L --> S at amino acid positions 59,114 and 155 respectively) in the Indian DENV-1 sequences, sequenced in this study. Except two 1997-98 Delhi isolates, which group in genotype I; all other Indian isolates group in genotype III. All Indian genotype III DENV-1 exhibited diversity among them, giving rise to at least 4 distinct lineages (India 1-4) showing proximity to isolates from diverse geographic locations. CONCLUSION: The extensive phylogenetic analysis revealed consistent existence of multiple lineages of DENV-1 genotype III during the last 5 decades in India. PMID- 19126195 TI - Intraoperative anaphylaxis due to gelofusine in a patient undergoing intramedullary nailing of the femur: a case report. AB - BACKGROUND: Although uncommon, anaphylaxis due to a colloid plasma expander can occur peri-operatively CASE PRESENTATION: We present a case of an intra-operative cardiac arrest in a 72 year old Caucasian male patient who underwent prophylactic intramedullary nailing for a proximal femoral metastasis from prostate cancer. The patient was resuscitated successfully and the procedure was completed uneventfully. Elevated serum tryptase levels confirmed the diagnosis of an anaphylactic reaction and positive allergy skin prick testing identified gelofusine as the causative agent. CONCLUSION: A high index of suspicion, prompt diagnosis and rapid institution of treatment are essential for a safe outcome following such reactions. To our knowledge, this is the first published report of such a severe reaction to gelofusine infusion that occurs during an orthopaedic procedure. PMID- 19126196 TI - Experiences with surgical treatment of ventricle septal defect as a post infarction complication. AB - BACKGROUND: Complications of acute myocardial infarction (AMI) with mechanical defects are associated with poor prognosis. Surgical intervention is indicated for a majority of these patients. The goal of surgical intervention is to improve the systolic cardiac function and to achieve a hemodynamic stability. In this present study we reviewed the outcome of patients with post infarction ventricular septal defect (PVSD) who underwent cardiac surgery. METHODS: We analysed retrospectively the hospital records of 41 patients, whose ages range from 48 to 81, and underwent a surgical treatment between 1990 and 2005 because of PVSD. RESULTS: In 22 patients concomitant coronary artery bypass grafting (CAGB) was performed. In 15 patients a residual shunt was found, this required re op in seven of them. The time interval from infarct to rupture was 8.7 days and from rupture to surgery was 23.1 days. Hospital mortality in PVSD group was 32%. The mortality of urgent repair within 3 days of intractable cardiogenic shock was 100%. The mortality of patients with an anterior VSD and a posterior VSD was 29.6% vs 42.8%, respectively. All patients who underwent the surgical repair later than day 36 survived. CONCLUSION: Surgical intervention is indicated for a majority of patients with mechanical complications. Cardiogenic shock remains the most important factor that affects the early results. The surgical repair of PVSD should be performed 4-5 weeks after AMI. To improve surgical outcome and hemodynamics the choice of surgical technique and surgical timing as well as preoperative management should be tailored for each patient individually. PMID- 19126197 TI - Vectors and transmission dynamics for Setaria tundra (Filarioidea; Onchocercidae), a parasite of reindeer in Finland. AB - BACKGROUND: Recent studies have revealed expansion by an array of Filarioid nematodes' into the northern boreal region of Finland. The vector-borne nematode, Setaria tundra, caused a serious disease outbreak in the Finnish reindeer population in 2003-05. The main aim of this study was to understand the outbreak dynamics and the rapid expansion of S. tundra in the sub arctic. We describe the vectors of S. tundra, and its development in vectors, for the first time. Finally we discuss the results in the context of the host-parasite ecology of S. tundra in Finland RESULTS: Development of S. tundra to the infective stage occurs in mosquitoes, (genera Aedes and Anopheles). We consider Aedes spp. the most important vectors. The prevalence of S. tundra naturally infected mosquitoes from Finland varied from 0.5 to 2.5%. The rate of development in mosquitoes was temperature-dependent. Infective larvae were present approximately 14 days after a blood meal in mosquitoes maintained at room temperature (mean 21 C), but did not develop in mosquitoes maintained outside for 22 days at a mean temperature of 14.1 C. The third-stage (infective) larvae were elongated (mean length 1411 mum (SD 207), and width 28 mum (SD 2)). The anterior end was blunt, and bore two liplike structures, the posterior end slight tapering with a prominent terminal papilla. Infective larvae were distributed anteriorly in the insect's body, the highest abundance being 70 larvae in one mosquito. A questionnaire survey revealed that the peak activity of Culicidae in the reindeer herding areas of Finland was from the middle of June to the end of July and that warm summer weather was associated with reindeer flocking behaviour on mosquito-rich wetlands. CONCLUSION: In the present work, S. tundra vectors and larval development were identified and described for the first time. Aedes spp. mosquitoes likely serve as the most important and competent vectors for S. tundra in Finland. Warm summers apparently promote transmission and genesis of disease outbreaks by favouring the development of S. tundra in its mosquito vectors, by improving the development and longevity of mosquitoes, and finally by forcing the reindeer to flock on mosquito rich wetlands. Thus we predict that global climate change has the potential to promote the further emergence of Filarioid nematodes and the disease caused by them in subarctic regions. PMID- 19126198 TI - Ascending cholangitis presenting with Lactococcus lactis cremoris bacteraemia: a case report. AB - INTRODUCTION: A case of Lactococcus lactis cremoris causing cholangitis is described. This Gram-positive organism is not routinely considered to be pathogenic in immunocompetent individuals. To our knowledge, this is the thirteenth report of invasive infection and the first of cholangitis to be reported in association with this organism. CASE PRESENTATION: A 72-year-old patient presented with Charcot's triad and was demonstrated to have cholangitis with Lactococcus lactis cremoris bacteraemia. Biliary drainage was achieved through endoscopic retrograde cholangiography. Antibiotic therapy with multiple agents was necessary. CONCLUSION: This report provides corroboration of evidence that Lactococcus lactis cremoris is a potential pathogen in immunocompetent adults. There remains a debate about the most appropriate empirical antibiotic therapy in this condition. In the light of this case, it is important to keep an open mind to potential pathogens. PMID- 19126199 TI - Quantification of fibronectin 1 (FN1) splice variants, including two novel ones, and analysis of integrins as candidate FN1 receptors in bovine preimplantation embryos. AB - BACKGROUND: Fibronectin 1 (FN1), a glycoprotein component of the extracellular matrix, exerts different functions during reproductive processes such as fertilisation, gastrulation and implantation. FN1 expression has been described to increase significantly from the morula towards the early blastocyst stage, suggesting that FN1 may also be involved in early blastocyst formation. By alternative splicing at 3 defined regions, different FN1 isoforms are generated, each with a unique biological function. The analysis of the alternative FN1 splicing on the one hand and the search for candidate FN1 receptors on the other hand during early bovine embryo development may reveal more about its function during bovine preimplantation embryo development. RESULTS: RT-qPCR quantification of the FN1 splice isoforms in oocytes, embryos, cumulus cells and adult tissue samples revealed a large variation in overall FN1 expression and in splice variant expression. Moreover, two new FN1 transcript variants were identified, the first one expressed in bovine preimplantation embryos and the second one expressed in cumulus cells. In the search for candidate receptors for the new embryo specific FN1 isoform, RNA expression analysis identified 5 alpha integrin subunits (ITGA2B, ITGA3, ITGA5, ITGA8, ITGAV) and 2 beta integrin subunits (ITGB1 and ITGB3) with a similar or overlapping RNA expression pattern as compared to FN1. But double immunofluorescent stainings could not confirm complete co localisation between FN1 and one out of 3 selected integrins alpha subunits (ITGA3, ITGA5, ITGAV). CONCLUSION: The existence of a new FN1 transcript variant, specifically expressed in morulae and blastocysts strengthens the idea that FN1 is involved in the process of compaction and blastocyst formation. Analysis of the integrin expression could not identify the binding partner for the embryo specific FN1 transcript variant making further steps necessary for the identification of the FN1 receptor and the downstream effects of FN1-receptor binding. PMID- 19126200 TI - Comparison of public peak detection algorithms for MALDI mass spectrometry data analysis. AB - BACKGROUND: In mass spectrometry (MS) based proteomic data analysis, peak detection is an essential step for subsequent analysis. Recently, there has been significant progress in the development of various peak detection algorithms. However, neither a comprehensive survey nor an experimental comparison of these algorithms is yet available. The main objective of this paper is to provide such a survey and to compare the performance of single spectrum based peak detection methods. RESULTS: In general, we can decompose a peak detection procedure into three consequent parts: smoothing, baseline correction and peak finding. We first categorize existing peak detection algorithms according to the techniques used in different phases. Such a categorization reveals the differences and similarities among existing peak detection algorithms. Then, we choose five typical peak detection algorithms to conduct a comprehensive experimental study using both simulation data and real MALDI MS data. CONCLUSION: The results of comparison show that the continuous wavelet-based algorithm provides the best average performance. PMID- 19126202 TI - Extramammary Paget's disease of the penis: a case report and review of the literature. AB - INTRODUCTION: Extramammary Paget's disease is a rare cutaneous, slow growing, intraepithelial adenocarcinoma developing in the apocrine gland-bearing areas. Isolated Paget's disease of the penis is extremely rare. CASE PRESENTATION: We describe the case of an 87-year-old Caucasian male who presented with a non healing erythematous plaque on the shaft of the penis previously misdiagnosed as Bowen's disease. The diagnosis was made histologically on the excised specimen and was supported by immunohistochemical staining. CONCLUSION: Extramammary Paget's disease is a rare disease which can mimic various types of dermatosis. A high index of suspicion is required, combined with biopsy and immunohistochemical staining in order to make the correct diagnosis. Long-term follow-up is mandatory in these patients in order to identify and treat any subsequent recurrence or concurrent malignancy. PMID- 19126201 TI - Rac1 regulates pancreatic islet morphogenesis. AB - BACKGROUND: Pancreatic islets of Langerhans originate from endocrine progenitors within the pancreatic ductal epithelium. Concomitant with differentiation of these progenitors into hormone-producing cells such cells delaminate, aggregate and migrate away from the ductal epithelium. The cellular and molecular mechanisms regulating islet cell delamination and cell migration are poorly understood. Extensive biochemical and cell biological studies using cultured cells demonstrated that Rac1, a member of the Rho family of small GTPases, acts as a key regulator of cell migration. RESULTS: To address the functional role of Rac1 in islet morphogenesis, we generated transgenic mice expressing dominant negative Rac1 under regulation of the Rat Insulin Promoter. Blocking Rac1 function in beta cells inhibited their migration away from the ductal epithelium in vivo. Consistently, transgenic islet cell spreading was compromised in vitro. We also show that the EGF-receptor ligand betacellulin induced actin remodelling and cell spreading in wild-type islets, but not in transgenic islets. Finally, we demonstrate that cell-cell contact E-cadherin increased as a consequence of blocking Rac1 activity. CONCLUSION: Our data support a model where Rac1 signalling controls islet cell migration by modulating E-cadherin-mediated cell cell adhesion. Furthermore, in vitro experiments show that betacellulin stimulated islet cell spreading and actin remodelling is compromised in transgenic islets, suggesting that betacellulin may act as a regulator of Rac1 activity and islet migration in vivo. Our results further emphasize Rac1 as a key regulator of cell migration and cell adhesion during tissue and organ morphogenesis. PMID- 19126203 TI - Dynamic simulations on the mitochondrial fatty acid beta-oxidation network. AB - BACKGROUND: The oxidation of fatty acids in mitochondria plays an important role in energy metabolism and genetic disorders of this pathway may cause metabolic diseases. Enzyme deficiencies can block the metabolism at defined reactions in the mitochondrion and lead to accumulation of specific substrates causing severe clinical manifestations. Ten of the disorders directly affecting mitochondrial fatty acid oxidation have been well-defined, implicating episodic hypoketotic hypoglycemia provoked by catabolic stress, multiple organ failure, muscle weakness, or hypertrophic cardiomyopathy. Additionally, syndromes of severe maternal illness (HELLP syndrome and AFLP) have been associated with pregnancies carrying a fetus affected by fatty acid oxidation deficiencies. However, little is known about fatty acids kinetics, especially during fasting or exercise when the demand for fatty acid oxidation is increased (catabolic stress). RESULTS: A computational kinetic network of 64 reactions with 91 compounds and 301 parameters was constructed to study dynamic properties of mitochondrial fatty acid beta-oxidation. Various deficiencies of acyl-CoA dehydrogenase were simulated and verified with measured concentrations of indicative metabolites of screened newborns in Middle Europe and South Australia. The simulated accumulation of specific acyl-CoAs according to the investigated enzyme deficiencies are in agreement with experimental data and findings in literature. Investigation of the dynamic properties of the fatty acid beta-oxidation reveals that the formation of acetyl-CoA - substrate for energy production - is highly impaired within the first hours of fasting corresponding to the rapid progress to coma within 1-2 hours. LCAD deficiency exhibits the highest accumulation of fatty acids along with marked increase of these substrates during catabolic stress and the lowest production rate of acetyl-CoA. These findings might confirm gestational loss to be the explanation that no human cases of LCAD deficiency have been described. CONCLUSION: In summary, this work provides a detailed kinetic model of mitochondrial metabolism with specific focus on fatty acid beta oxidation to simulate and predict the dynamic response of that metabolic network in the context of human disease. Our findings offer insight into the disease process (e.g. rapid progress to coma) and might confirm new explanations (no human cases of LCAD deficiency), which can hardly be obtained from experimental data alone. PMID- 19126204 TI - Frequency-specific modulation of population-level frequency tuning in human auditory cortex. AB - BACKGROUND: Under natural circumstances, attention plays an important role in extracting relevant auditory signals from simultaneously present, irrelevant noises. Excitatory and inhibitory neural activity, enhanced by attentional processes, seems to sharpen frequency tuning, contributing to improved auditory performance especially in noisy environments. In the present study, we investigated auditory magnetic fields in humans that were evoked by pure tones embedded in band-eliminated noises during two different stimulus sequencing conditions (constant vs. random) under auditory focused attention by means of magnetoencephalography (MEG). RESULTS: In total, we used identical auditory stimuli between conditions, but presented them in a different order, thereby manipulating the neural processing and the auditory performance of the listeners. Constant stimulus sequencing blocks were characterized by the simultaneous presentation of pure tones of identical frequency with band-eliminated noises, whereas random sequencing blocks were characterized by the simultaneous presentation of pure tones of random frequencies and band-eliminated noises. We demonstrated that auditory evoked neural responses were larger in the constant sequencing compared to the random sequencing condition, particularly when the simultaneously presented noises contained narrow stop-bands. CONCLUSION: The present study confirmed that population-level frequency tuning in human auditory cortex can be sharpened in a frequency-specific manner. This frequency-specific sharpening may contribute to improved auditory performance during detection and processing of relevant sound inputs characterized by specific frequency distributions in noisy environments. PMID- 19126205 TI - The first case of isolated facial cutanenous leishmaniasis in a Down syndrome infant: a case report and review of the literature. AB - BACKGROUND: Cutaneous leishmaniasis can be caused by several Leishmania species and is transmitted to human beings and animals by sand flies, Down syndrome is known to cause immunodeficiency that might lead to increase the susceptibility to infection with Leishmania.Up to our knowledge this is the first case of isolated facial cutaneous leshmaniasis in association with Down syndrome. CASE PRESENTATION: A 2 month old Saudi Arabian male infant was admitted in the pediatric ward of maternity and children's hospital, Buraidah, Kingdom of Saudi Arabia for the management of multiple ulcers on his face, two ulcers were big and were surrounded by edema, causing severe disfigurement. This disfigurement caused difficulty in recognizing the facial feature of Down syndrome. The presence of hypotonia, microcephaly, low set ears, bilateral simian creases and wide separation between big toe and other toes directed us to request karyotyping. The result of karyotyping confirmed the diagnosis of Down syndrome. CONCLUSION: Children with Down syndrome are immunodeficient, they have been reported to have a complex of immunological alterations which might lead to increased susceptibility to infection. PMID- 19126206 TI - Prenatal hypoxia induces increased cardiac contractility on a background of decreased capillary density. AB - BACKGROUND: Chronic hypoxia in utero (CHU) is one of the most common insults to fetal development and may be associated with poor cardiac recovery from ischaemia reperfusion injury, yet the effects on normal cardiac mechanical performance are poorly understood. METHODS: Pregnant female wistar rats were exposed to hypoxia (12% oxygen, balance nitrogen) for days 10-20 of pregnancy. Pups were born into normal room air and weaned normally. At 10 weeks of age, hearts were excised under anaesthesia and underwent retrograde 'Langendorff' perfusion. Mechanical performance was measured at constant filling pressure (100 cm H2O) with intraventricular balloon. Left ventricular free wall was dissected away and capillary density estimated following alkaline phosphatase staining. Expression of SERCA2a and Nitric Oxide Synthases (NOS) proteins were estimated by immunoblotting. RESULTS: CHU significantly increased body mass (P < 0.001) compared with age-matched control rats but was without effect on relative cardiac mass. For incremental increases in left ventricular balloon volume, diastolic pressure was preserved. However, systolic pressure was significantly greater following CHU for balloon volume = 50 microl (P < 0.01) and up to 200 microl (P < 0.05). For higher balloon volumes systolic pressure was not significantly different from control. Developed pressures were correspondingly increased relative to controls for balloon volumes up to 250 microl (P < 0.05). Left ventricular free wall capillary density was significantly decreased in both epicardium (18%; P < 0.05) and endocardium (11%; P < 0.05) despite preserved coronary flow. Western blot analysis revealed no change to the expression of SERCA2a or nNOS but immuno-detectable eNOS protein was significantly decreased (P < 0.001) in cardiac tissue following chronic hypoxia in utero. CONCLUSION: These data offer potential mechanisms for poor recovery following ischaemia, including decreased coronary flow reserve and impaired angiogenesis with subsequent detrimental effects of post-natal cardiac performance. PMID- 19126207 TI - Evaluation of recombinant invasive, non-pathogenic Eschericia coli as a vaccine vector against the intracellular pathogen, Brucella. AB - BACKGROUND: There is no safe, effective human vaccine against brucellosis. Live attenuated Brucella strains are widely used to vaccinate animals. However these live Brucella vaccines can cause disease and are unsafe for humans. Killed Brucella or subunit vaccines are not effective in eliciting long term protection. In this study, we evaluate an approach using a live, non-pathogenic bacteria (E. coli) genetically engineered to mimic the brucellae pathway of infection and present antigens for an appropriate cytolitic T cell response. METHODS: E. coli was modified to express invasin of Yersinia and listerialysin O (LLO) of Listeria to impart the necessary infectivity and antigen releasing traits of the intracellular pathogen, Brucella. This modified E. coli was considered our vaccine delivery system and was engineered to express Green Fluorescent Protein (GFP) or Brucella antigens for in vitro and in vivo immunological studies including cytokine profiling and cytotoxicity assays. RESULTS: The E. coli vaccine vector was able to infect all cells tested and efficiently deliver therapeutics to the host cell. Using GFP as antigen, we demonstrate that the E. coli vaccine vector elicits a Th1 cytokine profile in both primary and secondary immune responses. Additionally, using this vector to deliver a Brucella antigen, we demonstrate the ability of the E. coli vaccine vector to induce specific Cytotoxic T Lymphocytes (CTLs). CONCLUSION: Protection against most intracellular bacterial pathogens can be obtained mostly through cell mediated immunity. Data presented here suggest modified E. coli can be used as a vaccine vector for delivery of antigens and therapeutics mimicking the infection of the pathogen and inducing cell mediated immunity to that pathogen. PMID- 19126209 TI - Granulocytic sarcoma of the breast without development of bone marrow involvement: a case report. AB - A 29-year-old woman presented with a breast tumor with a primary diagnosis of MALT lymphoma. A repeat biopsy revealed a hematological neoplasm with diffuse, Indian file, and targetoid patterns. The cells were intermediate size with eosinophilic granules; the immunophenotyping showed monocytic differentiation, and no lymphoepithelial lesion was observed. The diagnosis was granulocytic sarcoma. Three different bone marrow biopsies were negative for neoplastic infiltration. After treatment, she developed secondary pancytopenia which contributed to her death 16 months after primary diagnosis. Granulocytic sarcoma of the breast is uncommon. A complete panel of immunohistochemistry is necessary to perform this diagnosis. PMID- 19126208 TI - Regeneration of the radial nerve cord in the sea cucumber Holothuria glaberrima. AB - BACKGROUND: Regeneration of neurons and fibers in the mammalian spinal cord has not been plausible, even though extensive studies have been made to understand the restrictive factors involved. New experimental models and strategies are necessary to determine how new nerve cells are generated and how fibers regrow and connect with their targets in adult animals. Non-vertebrate deuterostomes might provide some answers to these questions. Echinoderms, with their amazing regenerative capacities could serve as model systems; however, very few studies have been done to study the regeneration of their nervous system. RESULTS: We have studied nerve cord regeneration in the echinoderm Holothuria glaberrima. These are sea cucumbers or holothurians members of the class Holothuroidea. One radial nerve cord, part of the echinoderm CNS, was completely transected using a scalpel blade. Animals were allowed to heal for up to four weeks (2, 6, 12, 20, and 28 days post-injury) before sacrificed. Tissues were sectioned in a cryostat and changes in the radial nerve cord were analyzed using classical dyes and immunohistochemistry. In addition, the temporal and spatial distribution of cell proliferation and apoptosis was assayed using BrdU incorporation and the TUNEL assay, respectively.We found that H. glaberrima can regenerate its radial nerve cord within a month following transection. The regenerated cord looks amazingly similar in overall morphology and cellular composition to the uninjured cord. The cellular events associated to radial cord regeneration include: (1) outgrowth of nerve fibers from the injured radial cord stumps, (2) intense cellular division in the cord stumps and in the regenerating radial nerve cords, (3) high levels of apoptosis in the RNC adjacent to the injury and within the regenerating cord and (4) an increase in the number of spherule-containing cells. These events are similar to those that occur in other body wall tissues during wound healing and during regeneration of the intestine. CONCLUSION: Our data indicate that holothurians are capable of rapid and complete regeneration of the main component of their CNS. Regeneration involves both the outgrowth of nerve fibers and the formation of neurons. Moreover, the cellular events employed during regeneration are similar to those involved in other regenerative processes, namely wound healing and intestinal regeneration. Thus, holothurians should be viewed as an alternative model where many of the questions regarding nervous system regeneration in deuterostomes could be answered. PMID- 19126210 TI - Guillain-Barre Syndrome with asystole requiring permanent pacemaker: a case report. AB - INTRODUCTION: Guillain-Barre syndrome is an acute demyelinating disorder of the peripheral nervous system that results from an aberrant immune response directed at peripheral nerves. Autonomic abnormalities in Guillain-Barre syndrome are usually transient and reversible. We present a case of Guillain-Barre syndrome requiring a permanent pacemaker in view of persistent symptomatic bradyarrhythmia. CASE PRESENTATION: An 18-year-old Caucasian female presented with bilateral lower limb paraesthesias followed by bilateral progressive leg weakness and difficulty in walking. She reported an episode of an upper respiratory tract infection 3 weeks prior to the onset of her neurological symptoms. Diagnosis of Guillain-Barre syndrome was considered and a lumbar puncture was performed. Cerebrospinal fluid revealed albuminocytologic dissociation (increased protein but normal white blood cell count) suggestive of Guillain-Barre syndrome and hence an intravenous immunoglobulin G infusion was started. Within 48 hours, she progressed to complete flaccid quadriparesis with involvement of respiratory muscles requiring mechanical ventilatory support. Whist in the intensive care unit, she developed multiple episodes of bradycardia and asystole requiring a temporary pacemaker. In view of the persistent requirement for the temporary pacemaker for more than 5 days, she received a permanent pacemaker. She returned for follow-up three months after discharge with an intermittent need for ventricular pacing. CONCLUSION: Guillain-Barre syndrome can result in permanent damage to the cardiac conduction system. Patients with multiple episodes of bradycardia and asystole in the setting of Guillain-Barre syndrome should be evaluated and considered as potential candidates for permanent pacemaker implantation. PMID- 19126211 TI - Differential effects of 24-hydroxycholesterol and 27-hydroxycholesterol on beta amyloid precursor protein levels and processing in human neuroblastoma SH-SY5Y cells. AB - BACKGROUND: Activation of the liver x receptors (LXRs) by exogenous ligands stimulates the degradation of beta-amyloid 1-42 (Abeta42), a peptide that plays a central role in the pathogenesis of Alzheimer's disease (AD). The oxidized cholesterol products (oxysterols), 24-hydroxycholesterol (24-OHC) and 27 hydroxycholesterol (27-OHC), are endogenous activators of LXRs. However, the mechanisms by which these oxysterols may modulate Abeta42 levels are not well known. RESULTS: We determined the effect of 24-OHC and/or 27-OHC on Abeta generation in SH-SY5Y cells. We found that while 27-OHC increases levels of Abeta42, 24-OHC did not affect levels of this peptide. Increased Abeta42 levels with 27-OHC are associated with increased levels of beta-amyloid precursor protein (APP) as well as beta-secretase (BACE1), the enzyme that cleaves APP to yield Abeta. Unchanged Abeta42 levels with 24-OHC are associated with increased levels of sAPPalpha, suggesting that 24-OHC favors the processing of APP to the non-amyloidogenic pathway. Interestingly, 24-OHC, but not 27-OHC, increases levels of the ATP-binding cassette transporters, ABCA1 and ABCG1, which regulate cholesterol transport within and between cells. CONCLUSION: These results suggest that cholesterol metabolites are linked to Abeta42 production. 24-OHC may favor the non-amyloidogenic pathway and 27-OHC may enhance production of Abeta42 by upregulating APP and BACE1. Regulation of 24-OHC: 27-OHC ratio could be an important strategy in controlling Abeta42 levels in AD. PMID- 19126212 TI - Thermogenic effect of an acute ingestion of a weight loss supplement. AB - BACKGROUND: The purpose of this study was to examine the acute effects of a weight loss supplement on resting oxygen uptake (VO2), respiratory quotient (RQ), caloric expenditure (kcal), heart rate (HR), and blood pressure (BP) in healthy and physically active individuals. METHODS: Ten subjects (5 male, 5 female; 20.2 +/- 1.2 y; 172.2 +/- 8.9 cm; 71.5 +/- 17.2 kg; 17.3 +/- 2.6% body fat) underwent two testing sessions administered in a randomized and double-blind fashion. During each session, subjects reported to the Human Performance Laboratory after at least 3-h post-absorptive state and were provided either 3 capsules of the weight loss supplement (SUP), commercially marketed as Meltdown(R) or 3 capsules of a placebo (P). Subjects then rested in a semi-recumbent position for three hours. VO2 and HR were determined every 5 min during the first 30 min and every 10 min during the next 150 min. BP was determined every 15 min during the first 30 min and every 30 min thereafter. The profile of mood states was assessed every 30 min. RESULTS: Area under the curve analysis revealed a significant 28.9% difference in VO2 between SUP and P for the three hour study period. In addition, a significant difference in energy expenditure was also seen between SUP (1.28 +/ 0.33 kcal.min-1) and P (1.00 +/- 0.32 kcal.min-1). A trend (p = 0.06) towards a greater utilization of stored fat as an energy source was also demonstrated (0.78 +/- 0.23 kcal.min-1 and 0.50 +/- 0.38 kcal.min-1 in P and SUP, respectively). Significant elevations in HR were seen during hours two and three of the study, and significantly higher average systolic BP was observed between SUP (118.0 +/- 7.3 mmHg) and P (111.4 +/- 8.2 mmHg). No significant differences were seen in diastolic blood pressure at any time point. Significant increases in tension and confusion were seen in SUP. CONCLUSION: Results indicate a significant increase in energy expenditure in young, healthy individuals following an acute ingestion of a weight loss supplement. In addition, ingestion of this supplement appears modify mood and elevate HR and systolic BP following ingestion. PMID- 19126213 TI - Examination of a pre-exercise, high energy supplement on exercise performance. AB - BACKGROUND: The purpose of this study was to examine the effect of a pre-exercise high energy drink on reaction time and anaerobic power in competitive strength/power athletes. In addition, the effect of the pre-exercise drink on subjective feelings of energy, fatigue, alertness and focus was also explored. METHODS: Twelve male strength/power athletes (21.1 +/- 1.3 y; 179.8 +/- 7.1 cm; 88.6 +/- 12.1 kg; 17.6 +/- 3.3% body fat) underwent two testing sessions administered in a randomized and double-blind fashion. During each session, subjects reported to the Human Performance Laboratory and were provided with either 120 ml of a high energy drink (SUP), commercially marketed as Redline Extreme(R) or 120 ml of a placebo (PL) that was similar in taste and appearance but contained no active ingredients. Following consumption of the supplement or placebo subjects rested quietly for 10-minutes prior to completing a survey and commencing exercise. The survey consisted of 4 questions asking each subject to describe their feelings of energy, fatigue, alertness and focus for that moment. Following the completion of the questionnaire subjects performed a 2-minute quickness and reaction test on the Makoto testing device (Makoto USA, Centennial CO) and a 20-second Wingate Anaerobic Power test. Following a 10-minute rest subjects repeated the testing sequence and after a similar rest period a third and final testing sequence was performed. The Makoto testing device consisted of subjects reacting to both a visual and auditory stimulus and striking one out of 30 potential targets on three towers. RESULTS: Significant difference in reaction performance was seen between SUP and PL in both average number of targets struck (55.8 +/- 7.4 versus 51.9 +/- 7.4, respectively) and percent of targets struck (71.9 +/- 10.5% versus 66.8 +/- 10.9%, respectively). No significant differences between trials were seen in any anaerobic power measure. Subjective feelings of energy (3.5 +/- 0.5 versus 3.1 +/- 0.5) and focus (3.8 +/- 0.5 versus 3.3 +/- 0.7) were significantly higher during SUP compared to PL, respectively. In addition, a trend towards an increase in average alertness (p = 0.06) was seen in SUP compared to P. CONCLUSION: Results indicate a significant increase in reaction performance, with no effect on anaerobic power performance. In addition, ingestion of this supplement significantly improves subjective feelings of focus and energy in male strength/power athletes. PMID- 19126214 TI - Identification of suitable internal control genes for expression studies in Coffea arabica under different experimental conditions. AB - BACKGROUND: Quantitative data from gene expression experiments are often normalized by transcription levels of reference or housekeeping genes. An inherent assumption for their use is that the expression of these genes is highly uniform in living organisms during various phases of development, in different cell types and under diverse environmental conditions. To date, the validation of reference genes in plants has received very little attention and suitable reference genes have not been defined for a great number of crop species including Coffea arabica. The aim of the research reported herein was to compare the relative expression of a set of potential reference genes across different types of tissue/organ samples of coffee. We also validated the expression profiles of the selected reference genes at various stages of development and under a specific biotic stress. RESULTS: The expression levels of five frequently used housekeeping genes (reference genes), namely alcohol dehydrogenase (adh), 14 3-3, polyubiquitin (poly), beta-actin (actin) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) was assessed by quantitative real-time RT-PCR over a set of five tissue/organ samples (root, stem, leaf, flower, and fruits) of Coffea arabica plants. In addition to these commonly used internal controls, three other genes encoding a cysteine proteinase (cys), a caffeine synthase (ccs) and the 60S ribosomal protein L7 (rpl7) were also tested. Their stability and suitability as reference genes were validated by geNorm, NormFinder and BestKeeper programs. The obtained results revealed significantly variable expression levels of all reference genes analyzed, with the exception of gapdh, which showed no significant changes in expression among the investigated experimental conditions. CONCLUSION: Our data suggests that the expression of housekeeping genes is not completely stable in coffee. Based on our results, gapdh, followed by 14-3-3 and rpl7 were found to be homogeneously expressed and are therefore adequate for normalization purposes, showing equivalent transcript levels in different tissue/organ samples. Gapdh is therefore the recommended reference gene for measuring gene expression in Coffea arabica. Its use will enable more accurate and reliable normalization of tissue/organ-specific gene expression studies in this important cherry crop plant. PMID- 19126215 TI - Does doxycycline work in synergy with cisplatin and oxaliplatin in colorectal cancer? AB - BACKGROUND: In recent years, apart from antibacterial properties, doxycycline is reported to have cytotoxic and anti-proliferative actions in various cancers including colorectal cancer. Colorectal cancer constitutes one of the most common cancers in the western population. Apart from surgery, chemotherapy plays crucial role in the treatment of colorectal cancer. Cisplatin and oxaliplatin are most commonly used platinum compounds for the cancer chemotherapy. This study has looked for any impact of doxycycline on the cytotoxic effects of platinum compounds in colorectal cancer including its mechanisms of actions. METHODS: HT 29 colorectal cancer cells were used for this study. These cells were treated with cisplatin and oxaliplatin with or without doxycycline treatment. The caspase 3 gene expression was quantitated by gel electrophoresis and qualitated by real time polymerase chain reactions. The caspase 3 activity was assessed in HT 29 cells with fluorescence kit. RESULTS: The results revealed increased caspase 3 gene expressions and activities in HT 29 cells treated with cisplatin, oxaliplatin and doxycycline; however the combination of doxycycline with cisplatin and oxaliplatin did not report increased caspase 3 gene expressions and activity compared to cisplatin and oxaliplatin alone. CONCLUSION: We concluded that doxycycline has role in apoptosis induction in the colorectal cancer. However, it did not show any synergy with platinum compounds in the colorectal cancer cells. This study also pointed towards possible caspase-independent actions of doxycycline with cisplatin and oxaliplatin. However, further work is required to underpin the mechanisms of actions of doxycycline. PMID- 19126216 TI - Schizophrenia spectrum disorders in a Nigerian family: 4 case reports. AB - INTRODUCTION: The risk of developing schizophrenia is higher among persons with an affected sibling compared to the general population. Beliefs about aetiology influence presentation, management and treatment outcomes. There are no reports on multiple occurrences of this disorder in a family in this part of the world. In this case series we also highlight how cultural beliefs hamper and shape management. CASE PRESENTATION: We report a case series of schizophrenia spectrum disorders affecting four women in a Nigerian family of Ibo tribal origin who responded marginally to pharmacological interventions and required insight oriented psychotherapy and family therapy in resolving psychosocial problems. CONCLUSION: While western taught disease models might explain these presentations, it is not usually accepted by lay persons from developing cultures. Limitations in contemporary treatment approaches necessitate the development of culturally relevant psychotherapeutic interventions. PMID- 19126218 TI - The association of state per capita income and military service deaths in the Vietnam and Iraq wars. AB - BACKGROUND: In the United States, social burdens including war casualties are often distributed unequally across groups of individuals, communities, and states. The purpose of this report was to examine the association between war deaths and per capita income in the 50 states and District of Columbia during the Vietnam and Iraq wars. METHODS: The numbers of deaths by the home state of record for each conflict were obtained from Department of Defense records on the Internet as were key variables including age at death, gender, race, branch of service, rank, circumstances of death, home state of record and the ratio of wounded to dead. In addition, we obtained state per capita income and state population for the relevant times. RESULTS: Characteristics of decedents in the 2 conflicts were very similar with young, white enlisted men accounting for the majority of deaths. However, in the Iraq war, women accounted for a 2.4% of casualties. Also of note was the higher ratio of wounded to dead in Iraq. At the level of the state, the correlation between the ratio of deaths per 100,000 and per capita income was -0.51 (p < 0.0001) for Vietnam and -0.52 for Iraq (p < 0.0001). In both eras, states with lower per capita income tended to have higher ratios of deaths per population. CONCLUSION: For military service members serving in the Vietnam and Iraq conflicts, there were many more women who died in the latter war. Whether war deaths resulted in lower per capita income cannot be determined from these cross sectional data; we simply note a strong association between per capita income and war casualty rates for both wars. PMID- 19126217 TI - Bayesian optimal discovery procedure for simultaneous significance testing. AB - BACKGROUND: In high throughput screening, such as differential gene expression screening, drug sensitivity screening, and genome-wide RNAi screening, tens of thousands of tests need to be conducted simultaneously. However, the number of replicate measurements per test is extremely small, rarely exceeding 3. Several current approaches demonstrate that test statistics with shrinking variance estimates have more power over the traditional t statistic. RESULTS: We propose a Bayesian hierarchical model to incorporate the shrinkage concept by introducing a mixture structure on variance components. The estimates from the Bayesian model are utilized in the optimal discovery procedure (ODP) proposed by Storey in 2007, which was shown to have optimal performance in multiple significance tests. We compared the performance of the Bayesian ODP with several competing test statistics. CONCLUSION: We have conducted simulation studies with 2 to 6 replicates per gene. We have also included test results from two real datasets. The Bayesian ODP outperforms the other methods in our study, including the original ODP. The advantage of the Bayesian ODP becomes more significant when there are few replicates per test. The improvement over the original ODP is based on the fact that Bayesian model borrows strength across genes in estimating unknown parameters. The proposed approach is efficient in computation due to the conjugate structure of the Bayesian model. The R code (see Additional file 1) to calculate the Bayesian ODP is provided. PMID- 19126219 TI - A system analysis of a suboptimal surgical experience. AB - BACKGROUND: System analyses of incidents that occur in the process of health care delivery are rare. A case study of a series of incidents that one of the authors experienced after routine urologic surgery is presented. We interpret the sequence of events as a case of cascading incidents that resulted in outcomes that were suboptimal, although fortunately not fatal. METHODS: A system dynamics approach was employed to develop illustrative models (flow diagrams) of the dynamics of the patient's interaction with surgery and emergency departments. The flow diagrams were constructed based upon the experience of the patient, chart review, discussion with the involved physicians as well as several physician colleagues, comparison of our diagrams with those developed by the hospital of interest for internal planning purposes, and an iterative process with one of the co-authors who is a system dynamics expert. A dynamic hypothesis was developed using insights gained by building the flow diagrams. RESULTS: The incidents originated in design flaws and many small innocuous system changes that have occurred incrementally over time, which by themselves may have no consequence but in conjunction with some system randomness can have serious consequences. In the patient's case, the incidents that occurred in preoperative assessment and surgery originated in communication and procedural failures. System delays, communication failures, and capacity issues contributed largely to the subsequent incidents. Some of these issues were controllable by the physicians and staff of the institution, whereas others were less controllable. To the system's credit, some of the more controllable issues were addressed, but systemic problems like overcrowding are unlikely to be addressed in the near future. CONCLUSION: This is first instance that we are aware of in the literature where a system dynamics approach has been used to analyze a patient safety experience. The qualitative system dynamics analysis was useful in understanding the system, and contributed to learning on the part of some components of the system. We suggest that further data collection and quantitative analysis would be highly informative for identification of system changes to improve quality and safety. PMID- 19126220 TI - The Salmonella Pathogenicity Island (SPI) 1 contributes more than SPI2 to the colonization of the chicken by Salmonella enterica serovar Typhimurium. AB - BACKGROUND: Salmonella enterica serovar Typhimurium (Typhimurium) is an important pathogen that infects a broad range of hosts. In humans, Typhimurium causes a gastroenteritis characterized by vomiting, diarrhea, and abdominal pains. Typhimurium infection occurs mainly through the ingestion of contaminated food including poultry, pork, eggs, and milk. Chickens that are asymptomatic carriers of Typhimurium constitute a potential reservoir for infection. The type three secretion systems encoded by Salmonella pathogenicity islands (SPI) 1 and 2 are major virulence factors of Salmonella. However, only a few studies have investigated their role during the infection of chickens. RESULTS: We have taken a mixed infection approach to study the contribution of SPI1 and SPI2 to the colonization of the chicken by Typhimurium. We found that SPI1 contributes to colonization of both the cecum and spleen in the chicken. In contrast, SPI2 contributes to colonization of the spleen but not the cecum and, in the absence of SPI1, inhibits cecal colonization. Additionally, we show that the contribution of SPI1 in the spleen is greater than that of SPI2. These results are different from those observed during the infection of the mouse by Typhimurium where SPI2 is the major player during systemic colonization. CONCLUSION: The co-infection model we used provides a sensitive assay that confirms the role of SPI1 and clarifies the role of SPI2 in the colonization of the chicken by Typhimurium. PMID- 19126221 TI - Are figure legends sufficient? Evaluating the contribution of associated text to biomedical figure comprehension. AB - BACKGROUND: Biomedical scientists need to access figures to validate research facts and to formulate or to test novel research hypotheses. However, figures are difficult to comprehend without associated text (e.g., figure legend and other reference text). We are developing automated systems to extract the relevant explanatory information along with figures extracted from full text articles. Such systems could be very useful in improving figure retrieval and in reducing the workload of biomedical scientists, who otherwise have to retrieve and read the entire full-text journal article to determine which figures are relevant to their research. As a crucial step, we studied the importance of associated text in biomedical figure comprehension. METHODS: Twenty subjects evaluated three figure-text combinations: figure+legend, figure+legend+title+abstract, and figure+full-text. Using a Likert scale, each subject scored each figure+text according to the extent to which the subject thought he/she understood the meaning of the figure and the confidence in providing the assigned score. Additionally, each subject entered a free text summary for each figure-text. We identified missing information using indicator words present within the text summaries. Both the Likert scores and the missing information were statistically analyzed for differences among the figure-text types. We also evaluated the quality of text summaries with the text-summarization evaluation method the ROUGE score. RESULTS: Our results showed statistically significant differences in figure comprehension when varying levels of text were provided. When the full text article is not available, presenting just the figure+legend left biomedical researchers lacking 39-68% of the information about a figure as compared to having complete figure comprehension; adding the title and abstract improved the situation, but still left biomedical researchers missing 30% of the information. When the full-text article is available, figure comprehension increased to 86 97%; this indicates that researchers felt that only 3-14% of the necessary information for full figure comprehension was missing when full text was available to them. Clearly there is information in the abstract and in the full text that biomedical scientists deem important for understanding the figures that appear in full-text biomedical articles. CONCLUSION: We conclude that the texts that appear in full-text biomedical articles are useful for understanding the meaning of a figure, and an effective figure-mining system needs to unlock the information beyond figure legend. Our work provides important guidance to the figure mining systems that extract information only from figure and figure legend. PMID- 19126222 TI - Pancreatic and multiorgan resection with inferior vena cava reconstruction for retroperitoneal leiomyosarcoma. AB - BACKGROUND: Inferior vena cava (IVC) leiomyosarcoma is a rare tumor of smooth muscle origin. It is often large by the time of diagnosis and may involve adjacent organs. A margin-free resection may be curative, but the resection must involve the tumor en bloc with the affected segment of vena cava and locally involved organs. IVC resection often requires vascular reconstruction, which can be done with prosthetic graft. CASE PRESENTATION: We describe a 39-year-old man with an IVC leiomyosarcoma that involved the adrenal gland, distal pancreas, and blood supply to the spleen and left kidney. Tumor excision involved en bloc resection of all involved organs with reimplantation of the right renal vein and reconstruction of the IVC with a polytetrafluoroethylene graft. The patient recovered without renal insufficiency, graft infection, or other complications. Follow-up abdominal imaging at 1 year showed a patent IVC graft and no locally recurrent tumor. Prosthetic graft provides a sufficient diameter and length for replacement conduit in extensive resection of IVC leiomyosarcoma. CONCLUSION: To our knowledge, this is the first case of resection of an IVC sarcoma with prosthetic graft reconstruction in combination with pancreatic resection. Aggressive surgical resection including vascular reconstruction is warranted for select IVC tumors to achieve a potentially curative outcome. PMID- 19126223 TI - Fourmidable: a database for ant genomics. AB - BACKGROUND: Fourmidable is an infrastructure to curate and share the emerging genetic, molecular, and functional genomic data and protocols for ants. DESCRIPTION: The Fourmidable assembly pipeline groups nucleotide sequences into clusters before independently assembling each cluster. Subsequently, assembled sequences are annotated via Interproscan and BLAST against general and insect specific databases. Gene-specific information can be retrieved using gene identifiers, searching for similar sequences or browsing through inferred Gene Ontology annotations. The database will readily scale as ultra-high throughput sequence data and sequences from additional species become available. CONCLUSION: Fourmidable currently houses EST data from two ant species and microarray gene expression data for one of these. Fourmidable is publicly available at http://fourmidable.unil.ch. PMID- 19126224 TI - Predictive factors for somatization in a trauma sample. AB - BACKGROUND: Unexplained somatic symptoms are common among trauma survivors. The relationship between trauma and somatization appears to be mediated by posttraumatic stress disorder (PTSD). However, only few studies have focused on what other psychological risk factors may predispose a trauma victim towards developing somatoform symptoms. METHODS: The present paper examines the predictive value of PTSD severity, dissociation, negative affectivity, depression, anxiety, and feeling incompetent on somatization in a Danish sample of 169 adult men and women who were affected by a series of explosions in a firework factory settled in a residential area. RESULTS: Negative affectivity and feelings of incompetence significantly predicted somatization, explaining 42% of the variance. PTSD was significant until negative affectivity was controlled for. CONCLUSION: Negative affectivity and feelings of incompetence significantly predicted somatization in the trauma sample whereas dissociation, depression, and anxiety were not associated with degree of somatization. PTSD as a risk factor was mediated by negative affectivity. PMID- 19126225 TI - Carcinosarcoma of the breast: two case reports and review of the literature. AB - Carcinosarcoma of the breast, often referred to as metaplastic carcinoma of the breast, is a rare malignancy with two distinct cell lines described as a breast carcinoma of ductal type with a sarcoma-like component. Clinically, carcinosarcoma of the breast is an aggressive breast cancer. The prognosis for carcinosarcoma of the breast is less favorable compared to more common types of breast cancer such as infiltrating ductal or lobular carcinoma. Currently, the evaluation of breast carcinoma includes hormone receptor analysis of the tumor tissue, with those positive for estrogen or progesterone responding better to both hormonal and chemotherapy.Trastuzumab (Herceptin(R)) is available as an adjunct treatment for tumors which over-express the HER2/neu gene. Typically, metaplastic carcinomas of the breast do not express the estrogen or progesterone receptors and do not over-express the HER2/neu oncogene. As a result of this "triple negative" phenotype, such tumors tend to be more aggressive and are unlikely to respond to targeted therapy with Herceptin. The epidermal growth factor receptor HER-1/EGFR protein is expressed in the majority of metaplastic carcinomas and thus may serve as a potential therapeutic target for EGFR inhibitors such as gefitinib and cetuximab. The two cases we describe exemplify the aggressive nature of carcinosarcoma of the breast and support the findings that this tumor type does not express the common receptors found in other breast carcinomas. These case reports also emphasize the need for investigating the role for blockade of the HER-1/EGFR receptor with targeted therapies when found to be over-expressed in the primary tumor. PMID- 19126226 TI - Parallel shifts in ecology and natural selection in an island lizard. AB - BACKGROUND: Natural selection is a potent evolutionary force that shapes phenotypic variation to match ecological conditions. However, we know little about the year-to-year consistency of selection, or how inter-annual variation in ecology shapes adaptive landscapes and ultimately adaptive radiations. Here we combine remote sensing data, field experiments, and a four-year study of natural selection to show that changes in vegetation structure associated with a severe drought altered both habitat use and natural selection in the brown anole, Anolis sagrei. RESULTS: In natural populations, lizards increased their use of vegetation in wet years and this was correlated with selection on limb length but not body size. By contrast, a die-back of vegetation caused by drought was followed by reduced arboreality, selection on body size, and relaxed selection on limb length. With the return of the rains and recovery of vegetation, selection reverted back to pre-drought pattern of selection acting on limb length but not body size. To test for the impact of vegetation loss on natural selection during the drought, we experimentally removed vegetation on a separate study island in a naturally wet year. The experiment revealed similar inter-annual changes in selection on body size but not limb length. CONCLUSION: Our results illustrate the dynamic nature of ecology driving natural selection on Anolis morphology and emphasize the importance of inter-annual environmental variation in shaping adaptive variation. In addition, results illustrate the utility of using remote sensing data to examine ecology's role in driving natural selection. PMID- 19126227 TI - Automated alphabet reduction for protein datasets. AB - BACKGROUND: We investigate automated and generic alphabet reduction techniques for protein structure prediction datasets. Reducing alphabet cardinality without losing key biochemical information opens the door to potentially faster machine learning, data mining and optimization applications in structural bioinformatics. Furthermore, reduced but informative alphabets often result in, e.g., more compact and human-friendly classification/clustering rules. In this paper we propose a robust and sophisticated alphabet reduction protocol based on mutual information and state-of-the-art optimization techniques. RESULTS: We applied this protocol to the prediction of two protein structural features: contact number and relative solvent accessibility. For both features we generated alphabets of two, three, four and five letters. The five-letter alphabets gave prediction accuracies statistically similar to that obtained using the full amino acid alphabet. Moreover, the automatically designed alphabets were compared against other reduced alphabets taken from the literature or human-designed, outperforming them. The differences between our alphabets and the alphabets taken from the literature were quantitatively analyzed. All the above process had been performed using a primary sequence representation of proteins. As a final experiment, we extrapolated the obtained five-letter alphabet to reduce a, much richer, protein representation based on evolutionary information for the prediction of the same two features. Again, the performance gap between the full representation and the reduced representation was small, showing that the results of our automated alphabet reduction protocol, even if they were obtained using a simple representation, are also able to capture the crucial information needed for state-of-the-art protein representations. CONCLUSION: Our automated alphabet reduction protocol generates competent reduced alphabets tailored specifically for a variety of protein datasets. This process is done without any domain knowledge, using information theory metrics instead. The reduced alphabets contain some unexpected (but sound) groups of amino acids, thus suggesting new ways of interpreting the data. PMID- 19126228 TI - Intracellular amyloid formation in muscle cells of Abeta-transgenic Caenorhabditis elegans: determinants and physiological role in copper detoxification. AB - BACKGROUND: The amyloid beta-peptide is a ubiquitous peptide, which is prone to aggregate forming soluble toxic oligomers and insoluble less-toxic aggregates. The intrinsic and external/environmental factors that determine Abeta aggregation in vivo are poorly understood, as well as the cellular meaning of this process itself. Genetic data as well as cell biological and biochemical evidence strongly support the hypothesis that Abeta is a major player in the onset and development of Alzheimer's disease. In addition, it is also known that Abeta is involved in Inclusion Body Myositis, a common myopathy of the elderly in which the peptide accumulates intracellularly. RESULTS: In the present work, we found that intracellular Abeta aggregation in muscle cells of Caenorhabditis elegans overexpressing Abeta peptide is affected by two single amino acid substitutions, E22G (Arctic) and V18A (NIC). Both variations show decrease intracellular amyloidogenesis compared to wild type Abeta. We show that intracellular amyloid aggregation of wild type Abeta is accelerated by Cu2+ and diminished by copper chelators. Moreover, we demonstrate through toxicity and behavioral assays that Abeta-transgenic worms display a higher tolerance to Cu2+ toxic effects and that this resistance may be linked to the formation of amyloid aggregates. CONCLUSION: Our data show that intracellular Abeta amyloid aggregates may trap excess of free Cu2+ buffering its cytotoxic effects and that accelerated intracellular Abeta aggregation may be part of a cell protective mechanism. PMID- 19126229 TI - Patient complexity in quality comparisons for glycemic control: an observational study. AB - BACKGROUND: Patient complexity is not incorporated into quality of care comparisons for glycemic control. We developed a method to adjust hemoglobin A1c levels for patient characteristics that reflect complexity, and examined the effect of using adjusted A1c values on quality comparisons. METHODS: This cross sectional observational study used 1999 national VA (US Department of Veterans Affairs) pharmacy, inpatient and outpatient utilization, and laboratory data on diabetic veterans. We adjusted individual A1c levels for available domains of complexity: age, social support (marital status), comorbid illnesses, and severity of disease (insulin use). We used adjusted A1c values to generate VA medical center level performance measures, and compared medical center ranks using adjusted versus unadjusted A1c levels across several thresholds of A1c (8.0%, 8.5%, 9.0%, and 9.5%). RESULTS: The adjustment model had R2 = 8.3% with stable parameter estimates on thirty random 50% resamples. Adjustment for patient complexity resulted in the greatest rank differences in the best and worst performing deciles, with similar patterns across all tested thresholds. CONCLUSION: Adjustment for complexity resulted in large differences in identified best and worst performers at all tested thresholds. Current performance measures of glycemic control may not be reliably identifying quality problems, and tying reimbursements to such measures may compromise the care of complex patients. PMID- 19126230 TI - Whole brain radiotherapy with radiosensitizer for brain metastases. AB - PURPOSE: To study the efficacy of whole brain radiotherapy (WBRT) with radiosensitizer in comparison with WBRT alone for patients with brain metastases in terms of overall survival, disease progression, response to treatment and adverse effects of treatment. METHODS: A meta-analysis of randomized controlled trials (RCT) was performed in order to compare WBRT with radiosensitizer for brain metastases and WBRT alone. The MEDLINE, EMBASE, LILACS, and Cochrane Library databases, in addition to Trial registers, bibliographic databases, and recent issues of relevant journals were researched. Significant reports were reviewed by two reviewers independently. RESULTS: A total of 8 RCTs, yielding 2317 patients were analyzed. Pooled results from this 8 RCTs of WBRT with radiosensitizer have not shown a meaningful improvement on overall survival compared to WBRT alone OR = 1.03 (95% CI0.84-1.25, p = 0.77). Also, there was no difference in local brain tumor response OR = 0.8(95% CI 0.5 - 1.03) and brain tumor progression (OR = 1.11, 95% CI 0.9 - 1.3) when the two arms were compared. CONCLUSION: Our data show that WBRT with the following radiosentizers (ionidamine, metronidazole, misonodazole, motexafin gadolinium, BUdr, efaproxiral, thalidomide), have not improved significatively the overall survival, local control and tumor response compared to WBRT alone for brain metastases. However, 2 of them, motexafin- gadolinium and efaproxiral have been shown in recent publications (lung and breast) to have positive action in lung and breast carcinoma brain metastases in association with WBRT. PMID- 19126231 TI - Mucosa associated lymphoid tissue lymphoma presenting within a solitary anti mesenteric dilated segment of ileum: a case report. AB - INTRODUCTION: Mucosa associated lymphoid tissue (MALT) lymphoma is the third most common non-Hodgkin's lymphoma subtype. Clinical presentation is often insidious as a low-grade lesion and disease tends to remain localised for a long period of time. Ileal involvement is rare and presentation within an area of focal anti mesenteric ileal wall dilation simulating a large diverticulum has not been reported. CASE PRESENTATION: A 59-year-old man of Caucasian origin presented to a general surgical outpatients clinic with an 18-month history of intermittent upper abdominal pain following meals. Following normal gastroscopy and abdominal ultrasound, a focally dilated segment of ileum was seen on computed tomography and further clarified by barium investigation. Histology of this segment demonstrated MALT lymphoma of the small bowel. CONCLUSION: A solitary focally dilated segment of ileal wall may be neoplastic in nature and surgical resection needs to be considered. PMID- 19126232 TI - A procedure to correct proxy-reported weight in the National Health Interview Survey, 1976-2002. AB - BACKGROUND: Data from the National Health Interview Survey (NHIS) show a larger than-expected increase in mean BMI between 1996 and 1997. Proxy-reports of height and weight were discontinued as part of the 1997 NHIS redesign, suggesting that the sharp increase between 1996 and 1997 may be artifactual. METHODS: We merged NHIS data from 1976-2002 into a single database consisting of approximately 1.7 million adults aged 18 and over. The analysis consisted of two parts: First, we estimated the magnitude of BMI differences by reporting status (i.e., self reported versus proxy-reported height and weight). Second, we developed a procedure to correct biases in BMI introduced by reporting status. RESULTS: Our analyses confirmed that proxy-reports of weight tended to be biased downward, with the degree of bias varying by race, sex, and other characteristics. We developed a correction procedure to minimize BMI underestimation associated with proxy-reporting, substantially reducing the larger-than-expected increase found in NHIS data between 1996 and 1997. CONCLUSION: It is imperative that researchers who use reported estimates of height and weight think carefully about flaws in their data and how existing correction procedures might fail to account for them. The development of this particular correction procedure represents an important step toward improving the quality of BMI estimates in a widely used source of epidemiologic data. PMID- 19126233 TI - An HPV 16 L1-based chimeric human papilloma virus-like particles containing a string of epitopes produced in plants is able to elicit humoral and cytotoxic T cell activity in mice. AB - BACKGROUND: Even though two prophylactic vaccines against HPV are currently licensed, infections by the virus continue to be a major health problem mainly in developing countries. The cost of the vaccines limits wide-scale application in poor countries. A promising strategy for producing affordable and efficient vaccines involves the expression of recombinant immunogens in plants. Several HPV genes have been expressed in plants, including L1, which can self-assemble into virus-like particles. A plant-based, dual prophylactic/therapeutic vaccine remains an attractive possibility. RESULTS: We sought to express in tomato plants chimeric HPV 16 VLPs containing L1 fused to a string of epitopes from HPV 16 E6 and E7 proteins. The L1 employed had been modified to eliminate a strong inhibitory region at the 5' end of the molecule to increase expression levels. Several tomato lines were obtained expressing either L1 alone or L1-E6/E7 from 0.05% to 0.1% of total soluble protein. Stable integration of the transgenes was verified by Southern blot. Northern and western blot revealed successful expression of the transgenes at the mRNA and protein level. The chimeric VLPs were able to assemble adequately in tomato cells. Intraperitoneal administration in mice was able to elicit both neutralizing antibodies against the viral particle and cytotoxic T-lymphocytes activity against the epitopes. CONCLUSION: In this work, we report for the first time the expression in plants of a chimeric particle containing the HPV 16 L1 sequence and a string of T-cell epitopes from HPV 16 E6 and E7 fused to the C-terminus. The particles were able to induce a significant antibody and cytotoxic T-lymphocytes response. Experiments in vivo are in progress to determine whether the chimeric particles are able to induce regression of disease and resolution of viral infection in mice. Chimeric particles of the type described in this work may potentially be the basis for developing prophylactic/therapeutic vaccines. The fact that they are produced in plants, may lower production costs considerably. PMID- 19126234 TI - MRI bone oedema scores are higher in the arthritis mutilans form of psoriatic arthritis and correlate with high radiographic scores for joint damage. AB - INTRODUCTION: The aim of this study was to investigate the magnetic resonance imaging (MRI) features of bone disease in the arthritis mutilans (AM) form of psoriatic arthritis (PsA). METHODS: Twenty-eight patients with erosive PsA were enrolled (median disease duration of 14 years). Using x-rays of both hands and feet, 11 patients were classified as AM and 17 as non-AM (erosive psoriatic arthritis without bone lysis)by two observers. MRI scans (1.5T) of the dominant hand (wrist and fingers scanned separately) were obtained using standard contrast enhanced T1-weighted and fat-saturated T2-weighted sequences. Scans were scored separately by two readers for bone erosion, oedema and proliferation using a PsA MRI scoring system. X-rays were scored for erosions and joint space narrowing. RESULTS: On MRI, 1013 bones were scored by both readers. Reliability for scoring erosions and bone oedema was high (intraclass correlation coefficients = 0.80 and 0.77 respectively) but only fair for bone proliferation (intraclass correlation coefficient = 0.42). MRI erosion scores were higher in AM patients (53.0 versus 15.0, p = 0.004) as were bone oedema and proliferation scores (14.7 versus 10.0, p = 0.056 and 3.6 versus 0.7, p = 0.003 respectively). MRI bone oedema scores correlated with MRI erosion scores and X-ray erosion and joint space narrowing scores (r = 0.65, p = 0.0002 for all) but not the disease activity score 28-C reactive protein (DAS28CRP) or pain scores. CONCLUSIONS: In this patient group with PsA, MRI bone oedema, erosion and proliferation were all more severe in the AM-form. Bone oedema scores did not correlate with disease activity measures but were closely associated with X-ray joint damage scores. These results suggest that MRI bone oedema may be a pre-erosive feature and that bone damage may not be coupled with joint inflammation in PsA. PMID- 19126235 TI - A review of the benefits and risks of nonsteroidal anti-inflammatory drugs in the management of mild-to-moderate osteoarthritis. AB - This review is intended to provide physicians with an overview of the benefits and risks associated with the use of nonsteroidal anti-inflammatory drugs (NSAIDs) in the management of their patients with mild-to-moderate osteoarthritis (OA). New information on the inflammatory component of OA and the cardiovascular (CV) risk associated with cyclooxygenase (COX)-2-specific inhibitors has prompted efforts to revise the current recommendations for the use of NSAIDs in the treatment of patients with OA. Clinical studies have shown that naproxen and ibuprofen are significantly more effective at reducing OA pain than is acetaminophen, the traditional first-line therapy, which has no apparent anti inflammatory activity in the joints. The theoretical advantage of COX-2-specific inhibitors in reducing gastrointestinal (GI) toxicity has been demonstrated by clinical studies. GI complications can be reduced by using lower NSAID doses for the shortest duration or with a concomitant proton-pump inhibitor. All prescription NSAIDs carry a black box warning regarding CV risks; these risks vary among the NSAIDs. While ibuprofen and diclofenac are associated with an increased CV risk, naproxen was associated with a neutral CV risk relative to placebo. Ibuprofen, but not naproxen, attenuates the antiplatelet effects of aspirin. An understanding of the risks and benefits is important when choosing an NSAID. An exhaustive search of the medical literature since 1990 was conducted using the words "ibuprofen," "naproxen," "COX-2-specific NSAIDs," "nonspecific NSAIDs," "low-dose aspirin," and "nonprescription dosage." Databases searched included MEDLINE, EMBASE, and SCISEARCH. This article provides primary care physicians with the information needed to assist them in making more informed decisions in managing patients experiencing mild-to-moderate OA pain. PMID- 19126236 TI - Novel expression hosts for complex secondary metabolite megasynthetases: Production of myxochromide in the thermopilic isolate Corallococcus macrosporus GT-2. AB - Although many secondary metabolites with diverse biological activities have been isolated from myxobacteria, most strains of these biotechnologically important gliding prokaryotes remain difficult to handle genetically. In this study we describe the new fast growing myxobacterial thermophilic isolate GT-2 as a heterologous host for the expression of natural product biosynthetic pathways isolated from other myxobacteria. According to the results of sequence analysis of the 16S rDNA, this moderately thermophilic isolate is closely related to Corallococcus macrosporus and was therefore named C. macrosporus GT-2. Fast growth of moderately thermophilic strains results in shorter fermentation and generation times, aspects which are of significant interest for molecular biological work as well as production of secondary metabolites. Development of a genetic manipulation system allowed the introduction of the complete myxochromide biosynthetic gene cluster, located on a transposable fragment, into the chromosome of GT-2. Genetic engineering of the biosynthetic gene cluster by promoter exchange leads to much higher production of myxochromides in the heterologous host C. macrosporus GT-2 in comparison to the original producer Stigmatella aurantiaca and to the previously described heterologous host Pseudomonas putida (600 mg/L versus 8 mg/L and 40 mg/L, respectively). PMID- 19126238 TI - A deceptive MRI appearance of the medial meniscus in a 14 year old boy: a case report. AB - BACKGROUND: A 14 year old boy with a history and clinical examination suggestive of a medial meniscal tear is presented. CASE PRESENTATION: The MRI findings suggested a horizontal cleavage tear so arthroscopy was carried out. No intra articular pathology was found at the time of surgery. CONCLUSION: The role of MRI in investigation of meniscal injuries in children is discussed and the limitations highlighted. PMID- 19126237 TI - Association between risk factors for injurious falls and new benzodiazepine prescribing in elderly persons. AB - BACKGROUND: Benzodiazepines are frequently prescribed to elderly patients' despite concerns about adverse effects leading to injurious falls. Previous studies have not investigated the extent to which patients with pre-existing risk factors for falls are prescribed benzodiazepines. The objective of this study is to assess if some of the risk factors for falls are associated with new benzodiazepine prescriptions in elderly persons. METHODS: Using provincial administrative databases, elderly Quebec residents were screened in 1989 for benzodiazepine use and non-users were followed for up to 5 years. Logistic regression models were used to evaluate potential predictors of new benzodiazepine use among patient baseline characteristics. RESULTS: In the 252,811 elderly patients who had no benzodiazepine prescription during the baseline year (1989), 174,444 (69%) never filled a benzodiazepine prescription and 78,367 (31%) filled at least one benzodiazepine prescription. In the adjusted analysis, several risk factors for falls were associated with statistically significant increases in the risk of receiving a new benzodiazepine prescription including the number of prescribing physicians seen at baseline (OR: 1.12; 95% CI 1.11-1.13), being female (OR: 1.20; 95% CI 1.18-1.22) or a diagnosis of arthritis (OR: 1.11; 95% CI 1.09-1.14), depression (OR: 1.42; 95% CI 1.35-1.49) or alcohol abuse (OR: 1.24; 95% CI 1.05-1.46). The strongest predictor for starting a benzodiazepine was the use of other medications, particularly anti-depressants (OR: 1.85; 95% CI 1.75-1.95). CONCLUSION: Patients with pre-existing conditions that increase the risk of injurious falls are significantly more likely to receive a new prescription for a benzodiazepine. The strength of the association between previous medication use and new benzodiazepine prescriptions highlights an important medication safety issue. PMID- 19126239 TI - Water aerobics II: maternal body composition and perinatal outcomes after a program for low risk pregnant women. AB - BACKGROUND: To evaluate the effectiveness and safety of water aerobics during pregnancy. METHODS: A randomized controlled trial carried out in 71 low-risk sedentary pregnant women, randomly allocated to water aerobics or no physical exercise. Maternal body composition and perinatal outcomes were evaluated. For statistical analysis Chi-square, Fisher's or Student's t-tests were applied. Risk ratios and their 95% CI were estimated for main outcomes. Body composition was evaluated across time using MANOVA or Friedman multiple analysis. RESULTS: There were no significant differences between the groups regarding maternal weight gain, BMI or percentage of body fat during pregnancy. Incidence of preterm births (RR = 0.84; 95%CI:0.28-2.53), vaginal births (RR = 1.24; 95%CI:0.73-2.09), low birthweight (RR = 1.30; 95%CI:0.61-2.79) and adequate weight for gestational age (RR = 1.50; 95%CI:0.65-3.48) were also not significantly different between groups. There were no significant differences in systolic and diastolic blood pressure and heart rate between before and immediately after the water aerobics session. CONCLUSION: Water aerobics for sedentary pregnant women proved to be safe and was not associated with any alteration in maternal body composition, type of delivery, preterm birth rate, neonatal well-being or weight. PMID- 19126240 TI - Universal access: the benefits and challenges in bringing integrated HIV care to isolated and conflict affected populations in the Republic of Congo. AB - The Pool region of the Republic of Congo is an isolated, conflict-affected area with under-resourced and poorly functioning health care services. Despite significant AIDS-related mortality and morbidity in this area, and a national level commitment to universal HIV care, HIV has been largely neglected. In 2005 Medecins Sans Frontieres decided to introduce HIV care activities. However, in this setting of high basic health care needs, limited medical resources and competing medical priorities, a vertical HIV programme was not suitable. This paper describes the process of integrating HIV care and treatment into basic health services, the clinical outcomes of 222 patients started on antiretroviral treatment (ART), and the benefits to communities and health care systems. Key lessons learned include the use of multi-skilled human resources, the step-wise implementation of HIV activities, the initial engagement of an HIV experienced staff member, the use of simplified and adapted testing, clinical and monitoring protocols and drug regimens, the introduction of more complex monitoring tools to simplify clinical management decisions and intensive staff education regarding the benefits of HIV integration. This project in a rural and remote conflict affected setting demonstrates that integrated HIV programs can save lives and play a key role in helping to achieve universal access to ART in Africa. PMID- 19126241 TI - Role of acid-sensing ion channel 3 in sub-acute-phase inflammation. AB - BACKGROUND: Inflammation-mediated hyperalgesia involves tissue acidosis and sensitization of nociceptors. Many studies have reported increased expression of acid-sensing ion channel 3 (ASIC3) in inflammation and enhanced ASIC3 channel activity with pro-inflammatory mediators. However, the role of ASIC3 in inflammation remains inconclusive because of conflicting results generated from studies of ASIC3 knockout (ASIC3-/-) or dominant-negative mutant mice, which have shown normal, decreased or increased hyperalgesia during inflammation. RESULTS: Here, we tested whether ASIC3 plays an important role in inflammation of subcutaneous tissue of paw and muscle in ASIC3-/- mice induced by complete Freund's adjuvant (CFA) or carrageenan by investigating behavioral and pathological responses, as well as the expression profile of ion channels. Compared with the ASIC3+/+ controls, ASIC3-/- mice showed normal thermal and mechanical hyperalgesia with acute (4-h) intraplantar CFA- or carrageenan-induced inflammation, but the hyperalgesic effects in the sub-acute phase (1-2 days) were milder in all paradigms except for thermal hyperalgesia with CFA-induced inflammation. Interestingly, carrageenan-induced primary hyperalgesia was accompanied by an ASIC3-dependent Nav1.9 up-regulation and increase of tetrodotoxin (TTX)-resistant sodium currents. CFA-inflamed muscle did not evoke hyperalgesia in ASIC3-/- or ASIC3+/+ mice, whereas carrageenan-induced inflammation in muscle abolished mechanical hyperalgesia in ASIC3-/- mice, as previously described. However, ASIC3-/- mice showed attenuated pathological features such as less CFA-induced granulomas and milder carrageenan-evoked vasculitis as compared with ASIC3+/+ mice. CONCLUSION: We provide a novel finding that ASIC3 participates in the maintenance of sub-acute-phase primary hyperalgesia in subcutaneous inflammation and mediates the process of granuloma formation and vasculitis in intramuscular inflammation. PMID- 19126243 TI - Masked-Volume-Wise PCA and "reference Logan" illustrate similar regional differences in kinetic behavior in human brain PET study using [11C]-PIB. AB - BACKGROUND: Kinetic modeling using reference Logan is commonly used to analyze data obtained from dynamic Positron Emission Tomography (PET) studies on patients with Alzheimer's disease (AD) and healthy volunteers (HVs) using amyloid imaging agent N-methyl [11C]2-(4'-methylaminophenyl)-6-hydroxy-benzothiazole, [11C]-PIB. The aim of the present study was to explore whether results obtained using the newly introduced method, Masked Volume Wise Principal Component Analysis, MVW PCA, were similar to the results obtained using reference Logan. METHODS: MVW-PCA and reference Logan were performed on dynamic PET images obtained from four Alzheimer's disease (AD) patients on two occasions (baseline and follow-up) and on four healthy volunteers (HVs). Regions of interest (ROIs) of similar sizes were positioned in different parts of the brain in both AD patients and HVs where the difference between AD patients and HVs is largest. Signal-to-noise ratio (SNR) and discrimination power (DP) were calculated for images generated by the different methods and the results were compared both qualitatively and quantitatively. RESULTS: MVW-PCA generated images that illustrated similar regional binding patterns compared to reference Logan images and with slightly higher quality, enhanced contrast, improved SNR and DP, without being based on modeling assumptions. MVW-PCA also generated additional MVW-PC images by using the whole dataset, which illustrated regions with different and uncorrelated kinetic behaviors of the administered tracer. This additional information might improve the understanding of kinetic behavior of the administered tracer. CONCLUSION: MVW-PCA is a potential multivariate method that without modeling assumptions generates high quality images, which illustrated similar regional changes compared to modeling methods such as reference Logan. In addition, MVW PCA could be used as a new technique, applicable not only on dynamic human brain studies but also on dynamic cardiac studies when using PET. PMID- 19126242 TI - A study on the differential protein profiles in liver cells of heat stress rats with and without turpentine treatment. AB - BACKGROUND: Heat stress (HS) and related illnesses are a major concern in military, sports, and fire brigadiers. HS results in physiologic responses of increased temperature, heart rate and sweating. In heat stroke, inflammatory response plays an important role and it is evidenced that turpentine (T) induced circulating inflammatory cytokines reduced survival rate and duration at 42 degrees C. Here we report the alteration in the protein expression in liver cells upon HS with and without T treatment using two dimensional gel electrophoresis (2 DE), tryptic in-gel digestion and MALDI-TOF-MS/MS approaches. RESULTS: The effects of HS and T treatments alone and a combined treatments (T+HS) was performed in Wistar rat models. Proteomic analysis of liver in the HS and T+HS groups were analyzed compared to liver profiles of resting control and T treated groups. The study revealed a total of 25 and 29 differentially expressed proteins in the HS and T+HS groups respectively compared to resting control group. Fourteen proteins showed altered expression upon T treatment compared to resting control group. Proteins that are involved in metabolic and signal transduction pathways, defense, redox regulation, and cytoskeletal restructuring functions were identified. The altered expression of proteins reflected in 2D gels were corroborated by quantitative real time RT-PCR analysis of 8 protein coding genes representing metabolic and regulatory pathways for their expression and normalized with the house keeping gene beta-actin. CONCLUSION: The present study has identified a number of differentially expressed proteins in the liver cells of rats subjected to T, HS and T+HS treatments. Most of these proteins are implicated in cell metabolism, as well as adaptive response to incurred oxidative stress and tissue damage due to T+HS and HS effects. PMID- 19126244 TI - Frequent loss of heterozygosity and altered expression of the candidate tumor suppressor gene 'FAT' in human astrocytic tumors. AB - BACKGROUND: We had earlier used the comparison of RAPD (Random Amplification of Polymorphic DNA) DNA fingerprinting profiles of tumor and corresponding normal DNA to identify genetic alterations in primary human glial tumors. This has the advantage that DNA fingerprinting identifies the genetic alterations in a manner not biased for locus. METHODS: In this study we used RAPD-PCR to identify novel genomic alterations in the astrocytic tumors of WHO grade II (Low Grade Diffuse Astrocytoma) and WHO Grade IV (Glioblastoma Multiforme). Loss of heterozygosity (LOH) of the altered region was studied by microsatellite and Single Nucleotide Polymorphism (SNP) markers. Expression study of the gene identified at the altered locus was done by semi-quantitative reverse-transcriptase-PCR (RT-PCR). RESULTS: Bands consistently altered in the RAPD profile of tumor DNA in a significant proportion of tumors were identified. One such 500 bp band, that was absent in the RAPD profile of 33% (4/12) of the grade II astrocytic tumors, was selected for further study. Its sequence corresponded with a region of FAT, a putative tumor suppressor gene initially identified in Drosophila. Fifty percent of a set of 40 tumors, both grade II and IV, were shown to have Loss of Heterozygosity (LOH) at this locus by microsatellite (intragenic) and by SNP markers. Semi-quantitative RT-PCR showed low FAT mRNA levels in a major subset of tumors. CONCLUSION: These results point to a role of the FAT in astrocytic tumorigenesis and demonstrate the use of RAPD analysis in identifying specific alterations in astrocytic tumors. PMID- 19126245 TI - Assessing the impact of England's National Health Service R&D Health Technology Assessment program using the "payback" approach. AB - OBJECTIVES: This study assesses the impact of the English National Health Service (NHS) Health Technology Assessment (HTA) program using the "payback" framework. METHODS: A survey of lead investigators of all research projects funded by the HTA program 1993--2003 supplemented by more detailed case studies of sixteen projects. RESULTS: Of 204 eligible projects, replies were received from 133 or 65 percent. The mean number of peer-reviewed publications per project was 2.9. Seventy-three percent of projects claimed to have had had an impact on policy and 42 percent on behavior. Technology Assessment Reports for the National Institute for Health and Clinical Excellence (NICE) had fewer than average publications but greater impact on policy. Half of all projects went on to secure further funding. The case studies confirmed the survey findings and indicated factors associated with impact. CONCLUSIONS: The HTA program performed relatively well in terms of "payback." Facilitating factors included the program's emphasis on topics that matter to the NHS, rigorous methods and the existence of "policy customers" such as NICE. PMID- 19126246 TI - Use of surrogate outcomes in cost-effectiveness models: a review of United Kingdom health technology assessment reports. AB - OBJECTIVES: The aim of this study was to explore the use of surrogate outcomes--a substitute outcome that predicts final patient-related outcomes--in cost effectiveness models (CEM) within health technology assessment (HTA) reports and provide guidance for their future use. METHODS: Our sampling frame was all UK HTA Program monograph series reports published in 2005 and 2006. Reports were included if they addressed a treatment effectiveness/efficacy question and included a CEM based on a surrogate outcome. The two authors independently applied inclusion and exclusion criteria, and the following data was extracted from included reports: source of surrogate outcome, level of evidence for validation of the surrogate outcomes, methods used in report to quantify link between surrogate outcome and final outcome, and consideration of the uncertainty associated with using surrogate outcomes in the results or conclusions of report. RESULTS: Of 100 HTA reports, 35 complied with the inclusion criteria. Of these, four (11 percent) reports included a CEM based on a surrogate outcome. All four reports sourced treatment-related changes in surrogate outcome through a systematic review of the literature. One provided Level 1 surrogate evidence (randomized controlled trial data showing a strong association between the change in surrogate outcome and change final outcome); two reported Level 2 evidence (observational study data); and one provided Level 3 evidence (disease natural/ history data). The transparency of quantification and exploration of uncertainty of the surrogate and final outcome relationship varied considerably across all four reports. CONCLUSIONS: Recommendations are made for the use of surrogate outcomes in future HTA reports. PMID- 19126247 TI - Cost-effectiveness of magnetic resonance guided focused ultrasound for the treatment of uterine fibroids. AB - OBJECTIVES: The aim of this study is to evaluate the cost-effectiveness of Magnetic Resonance Guided Focused Ultrasound (MRgFUS) compared with alternative treatments for uterine fibroids in the United States. METHODS: We used techniques of decision analysis and data from secondary sources to develop and estimate an economic model of the management of uterine fibroids among premenopausal women. Patients in the model receive treatment with MRgFUS, uterine artery embolization (UAE), abdominal myomectomy, hysterectomy, or pharmacotherapy. The model predicts total costs (including subsequent procedures) and quality-adjusted life-years (QALYs) for each treatment strategy over a lifetime horizon, discounted at 3 percent, from a societal perspective. Data on treatment efficacy and safety were obtained from published and unpublished studies. Costs (2005 US$) were obtained from an analysis of a large administrative database and other secondary sources. Lost productivity costs were included in the base-case analysis, but excluded in a sensitivity analysis. RESULTS: UAE was associated with the most QALYs (17.39), followed by MRgFUS (17.36), myomectomy (17.31), hysterectomy (17.18), and pharmacotherapy (16.70). Pharmacotherapy was the least costly strategy ($9,200 per patient), followed by hysterectomy ($19,800), MRgFUS ($27,300), UAE ($28,900), and myomectomy ($35,100). Incremental cost-effectiveness ratios (cost per QALY gained) were $21,800 for hysterectomy, $41,400 for MRgFUS, and $54,200 for UAE; myomectomy was more costly and less effective than both MRgFUS and UAE. Results were sensitive to MRgFUS recurrence rates, MRgFUS procedure costs, and assumptions about quality of life following hysterectomy. CONCLUSIONS: Our findings suggest that MRgFUS is in the range of currently accepted criteria for cost-effectiveness, along with hysterectomy and UAE. PMID- 19126248 TI - An economic analysis of continuous positive airway pressure for the treatment of obstructive sleep apnea-hypopnea syndrome. AB - OBJECTIVES: An important option for the medical treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS) is continuous positive airway pressure (CPAP) during sleep. This study reports on the cost-effectiveness of CPAP compared with dental devices and lifestyle advice. The work was commissioned by the NHS HTA Programme to inform the National Institute of Health and Clinical Excellence's (NICE) appraisal of CPAP. METHODS: A Markov model compared the interventions over the expected patient lifetime. The primary measure of cost-effectiveness was the incremental cost per quality-adjusted life-year (QALY) gained. The QALY incorporated the impact of treatments on daytime sleepiness, blood pressure and health-related quality of life (HRQoL). RESULTS: On average, CPAP was associated with higher costs and QALYs compared with dental devices or lifestyle advice. In the base-case analysis, the incremental cost-effectiveness ratio (ICER) for CPAP compared with dental devices was around 4,000 pounds per QALY (2005--06 prices). The probability that CPAP is more cost-effective than dental devices or lifestyle advice at a threshold value of 20,000 pounds per QALY was 0.78 for men and 0.80 for women. Several sensitivity analyses were undertaken and it was found that the ICER for CPAP consistently fell below 20,000 pounds per QALY gained, apart from in a subgroup with mild disease. CONCLUSIONS: The model suggests that CPAP is cost-effective compared with dental devices and lifestyle advice for adults with moderate or severe symptomatic OSAHS at the cost-effectiveness thresholds used by NICE. This finding is reflected in the NICE guidance. PMID- 19126249 TI - Identifying patient-relevant endpoints among individuals with schizophrenia: an application of patient-centered health technology assessment. AB - OBJECTIVES: Schizophrenia imposes a great burden on society, and while evaluation should play an important role in informing society's efforts to alleviate these burdens, it is unclear what "endpoints" should be chosen as the objective of such analyses. The objectives of the study were to elicit endpoints directly from patients with schizophrenia, to ascertain whether patients are sufficiently cognoscente to express what endpoints are and are not important to them and to rank the relevant endpoints. METHODS: We applied principles of patient-centered health technology assessment to identify and value endpoints from the patient's perspective. Focus groups were conducted to elicit endpoints, using interpretive phenomalogical analysis (IPA) to guide the collection, analysis and interpretation of data. Patient interviews were subsequently used to elicit patient preference over endpoints. Respondents were presented with cards outlining the endpoints and asked to remove irrelevant cards. They where then asked to identify and rank their five most relevant endpoints in order of importance. Interviews were recorded for the purposed of triangulation, and data was analyzed using descriptive statistics. Patients were recruited from five geographically diverse cities in Germany. Eligibility required a diagnosis of schizophrenia by a physician and treatment with an antipsychotic medication for at least one year. Respondents were excluded if they were experiencing an acute episode. RESULTS: Thirteen endpoints emerged as important from the focus groups spanning side-effects, functional status, processes of care and clinical outcomes. Respondents could clearly identify relevant and irrelevant endpoints, and rank which factors were important to them. Triangulation between field notes of the ranking exercise and recordings confirmed that rankings were not arbitrary, but justified from the respondents' point of view. CONCLUSIONS: Patients with schizophrenia can express preferences over endpoints. Our results show that qualitative methods such as IPA can be used to identify factors, but ranking exercises provide a more robust method for ranking the importance of endpoints. Future research involving patients with schizophrenia ranking outcomes is needed to identify variations across patients and methods such as conjoint analysis could prove beneficial in identifying acceptable tradeoffs across endpoints. PMID- 19126250 TI - Assessment of the quality of mini-HTA. AB - OBJECTIVES: Mini-HTA (health technology assessment) is increasingly being applied in Denmark as an input for decisions on the use of health technologies. Mini-HTA is a form or check list with questions concerning the prerequisites for and consequences of health technologies. At the national level, the National Board of Health uses mini-HTA when hospitals apply for permission to introduce new treatments. Mini-HTA is also compulsory in Danish Regions' annual collection of early warnings. At the local level some hospitals have made mini-HTA compulsory when clinical departments apply for funding for new technologies. The objective of this study is to assess the quality of the information included in mini-HTA used at Danish hospitals and to discuss the consequences of this to decision making. METHODS: The quality of mini-HTA is assessed by use of an INATHA checklist for HTA reports. Data consists of reviews of the quality in fifty-two mini-HTAs produced by Danish hospitals in 2008. RESULTS: The mini-HTAs generally include descriptions of the assessed technology and the comparator, but information about the selection and interpretation of the clinical literature and other data is often missing. The level of evidence for the clinical effects and the main references are generally included. Only 25 percent of the mini-HTAs include a quantitative estimate of the size of the clinical effects. Organizational consequences inside the clinical department is described in 81percent of the cases and 92 percent includes a cost estimate. CONCLUSIONS: The results show that the quality of the information in many cases is insufficient. There is a strong need for quality assurance of mini-HTAs to improve the accuracy of the information, however, without harming the timeliness and the limited use of resources in producing the reports. PMID- 19126251 TI - Finding legitimacy for the role of budget impact in drug reimbursement decisions. AB - OBJECTIVES: Research has shown that effectiveness, cost-effectiveness, and severity of illness each play a role in drug reimbursement decisions. However, the role of budget impact in such decisions is less obvious. Policy makers almost always demand a budget impact estimate yet seem reluctant to formally include budget impact as a rationing criterion. Health economists even reject budget impact as a legitimate criterion. For these reasons, it is important to examine its use in rationing decisions, and rationales underlying its use. METHODS: We trace several rationales supporting the use of budget impact through a literature review, supplemented by semistructured interviews with eleven key stakeholders involved in drug reimbursement decisions in the Netherlands. RESULTS: Budget impact arguments are used in certain instances, although policy makers appear uncomfortable with its use because well described rationales still are lacking. In addition, we identify the following rationales to support budget impact as a rationing criterion: opportunity costs, loss aversion, uncertainty and equal opportunity. CONCLUSIONS: Budget impact plays a role in drug reimbursement decisions and has rationales to support its use. However, policy makers do not easily admit that they consider budget impact and are even reluctant to explicitly use budget impact as a formal criterion. A debate would strengthen the theoretical foundation of budget impact as a legitimate criterion in the context of drug reimbursement decisions. Such discussion of budget impact's role will also enhance policy-makers' accountability. PMID- 19126252 TI - Pegylated and non-pegylated interferon-alfa and ribavirin for the treatment of mild chronic hepatitis C: a systematic review and meta-analysis. AB - OBJECTIVES: Traditionally, patients with chronic hepatitis C virus (HCV) infection have not received treatment until their infection reaches the moderate to severe stage. The aim of this systematic review was to assess the clinical effectiveness of pegylated (PEG) and non-pegylated interferon (IFN) alfa and ribavirin (RBV) for the treatment of adults with histologically mild HCV. METHODS: We performed a sensitive search of fourteen electronic bibliographic databases for literature that met criteria defined in a research protocol. Two reviewers independently selected studies, extracted data and assessed methodological quality. RESULTS: Ten randomized, controlled trials (RCTs) were included. Treatment with PEG + RBV combination therapy resulted in significantly higher sustained virological response (SVR) rates than treatment with IFN + RBV combination therapy. Treatment for 48 weeks with PEG + RBV was significantly more effective than the same treatment for 24 weeks. Significantly higher SVR rates were seen with IFN + RBV compared with either IFN monotherapy or no treatment. In the meta-analysis (four IFN trials), the relative risk of not experiencing an SVR was 0.59 (95 percent CI, 0.51 - 0.69) and was statistically significant (p < .00001). SVRs were higher for patients with genotype non-1 compared with genotype 1 for both PEG + RBV and IFN + RBV treatments. CONCLUSIONS: Patients with histologically mild HCV can be successfully treated with both PEG and IFN combination therapy, and response rates are broadly comparable with those achieved in patients with advanced disease. Treating patients in the early milder stages of HCV is, therefore, a clinically effective option. PMID- 19126253 TI - Arthroplasty registers: a review of international experiences. AB - OBJECTIVES: Registers have proven to be a valuable instrument in the evaluation of arthroplasty procedures and the performance of implants. The aim of this study was to describe the structure, functioning, and content of arthroplasty registers in Europe and other parts of the world. METHODS: A search of technical reports was carried out through the Internet and in Medline/PubMed. The exhaustiveness of the information was confirmed using the links to Web pages of other registers and contacts with key people. Aims, methods in data collection and evaluation, internal structure and organization, participants, validity of the data, and other variables were assessed for each arthroplasty register using a qualitative content analysis of the texts. RESULTS: Fifteen arthroplasty registers were identified which published sufficient information to conduct a comparative analysis. Eight additional registers were identified but no information was available on the Internet or in English. Most registers were initiatives of an orthopaedic society receiving governmental funding. Data were collected using standardized clinical forms and additional information from clinical administrative datasets or other registers (mortality, implant costs, hip fractures). The main outcome measure of these registers is survival of the prostheses. Registers use the Internet and their annual reports as the main strategy for the dissemination and feed-back of their results. CONCLUSIONS: Scientific or professional societies and the public health administration should collaborate in the development of arthroplasty registers. To adequately assess the results of observational data information on the structure, the process of arthroplasty interventions and patients characteristics should be collected. PMID- 19126254 TI - Constructive Technology Assessment (CTA) as a tool in coverage with evidence development: the case of the 70-gene prognosis signature for breast cancer diagnostics. AB - OBJECTIVES: Constructive Technology Assessment (CTA) is a means to guide early implementation of new developments in society, and can be used as an evaluation tool for Coverage with Evidence Development (CED). We used CTA for the introduction of a new diagnostic test in the Netherlands, the 70-gene prognosis signature (MammaPrint) for node-negative breast cancer patients. METHODS: Studied aspects were (organizational) efficiency, patient-centeredness and diffusion scenarios. Pre-post structured surveys were conducted in fifteen community hospitals concerning changes in logistics and teamwork as a consequence of the introduction of the 70-gene signature. Patient-centeredness was measured by questionnaires and interviews regarding knowledge and psychological impact of the test. Diffusion scenarios, which are commonly applied in industry to anticipate on future development and diffusion of their products, have been applied in this study. RESULTS: Median implementation-time of the 70-gene signature was 1.2 months. Most changes were seen in pathology processes and adjuvant treatment decisions. Physicians valued the addition of the 70-gene signature information as beneficial for patient management. Patient-centeredness (n = 77, response 78 percent): patients receiving a concordant high-risk and discordant clinical low/high risk-signature showed significantly more negative emotions with respect to receiving both test-results compared with concordant low-risk and discordant clinical high/low risk-signature patients. The first scenario was written in 2004 before the introduction of the 70-gene signature and identified hypothetical developments that could influence diffusion; especially the "what-if" deviation describing a discussion on validity among physicians proved to be realistic. CONCLUSIONS: Differences in speed of implementation and influenced treatment decisions were seen. Impact on patients seems especially related to discordance and its successive communication. In the future, scenario drafting will lead to input for model-based cost-effectiveness analysis. Finally, CTA can be useful as a tool to guide CED by adding monitoring and anticipation on possible developments during early implementation, to the assessment of promising new technologies. PMID- 19126255 TI - Evaluating the status of "translating research into practice" at a major academic healthcare system. AB - OBJECTIVES: The aim of this study was to assess the status of translating research findings into practice at a major academic healthcare system in Central Texas. METHODS: We conducted a cross-sectional survey addressing knowledge of and participation in translational research of physicians, residents, nurses and third- and fourth-year medical students in a major academic healthcare system in Central Texas. RESULTS: Out of 508 respondents, 428 (84.3 percent) completed all questions. A total of 68.9 percent of faculty reported having sufficient education and training to conduct research versus 44.4 percent of residents and 35.6 percent of nurses. Fifty-eight percent of faculty, 53 percent of residents and 9 percent of nurses reported current involvement in research activity. A total of 55.6 percent of residents reported that their departments provide them with protected time for research versus 18.4 percent of faculty and 10.3 percent of nurses. In addition, 33.9 percent of nurses reported interest in participating in research but do not know how to start. There were 86.4 percent of faculty, 77.8 percent of residents, and 58 percent of nurses who indicated they were familiar with translational research. However, only 42.7 percent of faculty, 46.7 percent of residents and 35.6 percent of nurses indicated they were aware of any changes in the delivery of care that resulted from research projects. CONCLUSIONS: The study results suggested failure to leverage members of the healthcare team in a systematic process to ensure translation of research findings into practice. Results highlighted the need to merge culture of safety and quality improvement with research while dealing with the daily pressures of patient care. PMID- 19126256 TI - Health-related quality of life measures in routine clinical care: can FACT fatigue help to assess the management of fatigue in cancer patients? AB - OBJECTIVES: Fatigue is the most common symptom reported by cancer patients. The inclusion of health-related quality of life (HRQL) measures in routine clinical care of cancer patients may improve the management of fatigue. The primary objective of this study is to provide evidence on the magnitude of change in fatigue subscale scores using the Functional Assessment of Cancer Therapy-Fatigue (FACT-F) that is clinically important. METHODS: Consecutive patients with advanced primary lung cancer attending a Canadian tertiary care cancer and, prior to undergoing palliative chemotherapy, were enrolled in the study. Patients completed a battery of questionnaires [FACT-F, Qualitative Patients Self-report of Fatigue Level (QPSRF)] at baseline, follow-up and 2 weeks after their final cycle of chemotherapy. Clinicians assessed the patients using the Eastern Cooperative Oncology Group (ECOG) Performance Status Scale at baseline and each follow-up visit. FACT-F change scores were computed as the mean change in score (end of study score minus baseline score). RESULTS: A total of 43 patients with mean age of 59 years were enrolled in the study. Results revealed a mean change in FACT-F subscale score of 5.0 (SE 1.06) for those who rated themselves as more tired, 1.28 (SE 1.00) for those who rated themselves as the same (no change), and -1.52 (SE 0.84) for those patients who rated themselves as less tired. CONCLUSIONS: We provide evidence on the magnitude of change in FACT-F score that is associated with the perception by patients of improvement in fatigue and magnitude of change in score that is associated with worsening in fatigue. PMID- 19126257 TI - Information technology capacities assessment tool in hospitals: instrument development and validation. AB - OBJECTIVES: This research integrates existing literature on information technology (IT) in hospitals, and proposes and validates a comprehensive IT capacities assessment tool in these settings. METHODS: A comprehensive literature review was conducted on Medline until September 2006 to identify studies that used specific IT measures in hospitals. The results were mapped and used as a basis for the development of the proposed instrument, which was tested through a survey of Canadian healthcare organizations (N = 221). RESULTS: A total of seventeen studies provided indicators of clinical and administrative IT capacities in hospitals. Based on the mapping of these indicators, a comprehensive IT capacities assessment instrument was developed including thirty four items exploring computerized processes, thirteen items assessing contemporary technologies, and eleven items investigating internal and external information sharing. A time frame was inserted in the tool to reflect "plans for" versus "current" implementation of IT; in the latter, the extent of current use of computerized processes and technologies was measured on a (1-7) scale. Overall, the survey yielded a total of 106 responses (52.2 percent response rate), and the results demonstrated a good level of reliability and validity of the instrument. CONCLUSIONS: This study unifies existing work in this area, and presents the psychometric properties of an IT capacities assessment tool in hospitals. By developing scores for capturing IT capacities in hospitals, it is possible to further address important research questions related to the determinants and impacts of IT sophistication in these settings. PMID- 19126258 TI - The influence of methodologic quality on the conclusion of a landmark meta analysis on thrombolytic therapy. PMID- 19126259 TI - Colorectal cancer screening policy in Hungary. PMID- 19126261 TI - Cocoa: antioxidant and immunomodulator. AB - Cocoa, a product consumed since 600 BC, is now a subject of increasing interest because of its antioxidant properties, which are mainly attributed to the content of flavonoids such as ( - )-epicatechin, catechin and procyanidins. Moreover, recent findings suggest a regulatory effect of cocoa on the immune cells implicated in innate and acquired immunity. Cocoa exerts regulatory activity on the secretion of inflammatory mediators from macrophages and other leucocytes in vitro. In addition, emerging data from in vivo studies support an immunomodulating effect. Long-term cocoa intake in rats affects both intestinal and systemic immune function. Studies in this line suggest that high-dose cocoa intake in young rats favours the T helper 1 (Th1) response and increases intestinal gammadelta T lymphocyte count, whereas the antibody-secreting response decreases. The mechanisms involved in this activity are uncertain; nonetheless, because redox-sensitive pathways control immune cell function, the action of cocoa flavonoids on modulating cell signalling and gene expression deserves investigation. PMID- 19126262 TI - Conference on "Multidisciplinary approaches to nutritional problems". Postgraduate Symposium. The role of fat in gastric emptying and satiety: acute and chronic effects. AB - Dietary fat is an important factor in the aetiology of obesity and the metabolic syndrome. It has been widely debated whether gastric emptying (GE) is altered in obesity. GE times have been reported as both longer and shorter in obese individuals compared with matched lean individuals. However, the general consensus is that GE is accelerated and satiety is lower in obesity. Research has implicated a high-fat (HF) diet in these findings. A single HF meal has a longer GE time than a low-fat meal and can even delay GE of the subsequent meal. However, an HF diet has shown different effects. Feeding a HF diet adapts gastrointestinal function to reduce GE times in comparison with a low-fat diet. Increased GE may lead to decreased satiety and faster onset of subsequent eating episodes. Further results have suggested that consuming an HF diet for 14 d increases the GE rate of HF food but not low-fat food. Consuming HF diets for 2 weeks has also been shown to increase food intake. Decreased satiation following an HF diet may cause increased food intake and a positive energy balance, potentially resulting in a gradual increase in adiposity. Recent results have suggested that gastrointestinal transit is accelerated following only 3 d on a HF diet. The variable GE times reported in obesity may be associated with interactions between the HF diet and obesity and not simply the obese state. PMID- 19126263 TI - Serotonin transporter genotype is associated with cognitive performance but not regional 5-HT1A receptor binding in humans. AB - The human serotonin transporter (5-HTT) gene is one of the most extensively studied in psychiatry. A functional polymorphism in the promoter region of the 5 HTT gene (5-HTTLPR) has been associated with several psychiatric disorders as well as anxiety-related personality traits. In search of a mechanistic understanding of the functional implications of 5-HTTLPR, the influence of this polymorphism on regional 5-HT1A receptor density has previously been examined in two positron emission tomography (PET) studies in humans, yielding, however, contradictory results. In the present study, 54 control subjects were examined with [11C]WAY 100635 PET and a battery of cognitive tests. Regional binding potential (BP) of [11C]WAY 100635 to 5-HT1A receptor was calculated for the dorsal raphe nuclei, the hippocampus, the anterior cingulate, the insula, the temporal cortex and the frontal cortex. The influence of 5-HTTLPR genotype on regional 5-HT1A BP and cognitive performance was investigated. No differences in 5-HT1A receptor density between carriers and non-carriers of the S allele were found. Thus, we could not replicate any of the previously reported associations between 5-HTTLPR and 5-HT1A density. There was, however, a highly significant association between 5-HTTLPR genotype and performance in Wisconsin Card Sorting Test; carriers of the S allele had a superior performance compared to the LL carriers. These observations suggest that functional implications of the 5-HTTLPR polymorphism are not likely to be mediated by differences in 5-HT1A expression levels and that other biomarkers must be considered for future investigations at phenotype level. PMID- 19126264 TI - 'Treatment of bipolar disorder: a systematic review of available data and clinical perspectives' by Fountoulakis & Vieta (2008). PMID- 19126265 TI - Cumulus cell features and nuclear chromatin configuration of in vitro matured canine COCs and the influence of in vivo serum progesterone concentrations of ovary donors. AB - Phenotype integrity is viewed as an indicator of cumulus-oocyte complex (COC) viability. The objectives of this study were: (a) to observe the influence of cumulus investment expansion on the nuclear chromatin configuration of canine oocytes matured in vitro; (b) to examine the relationship between cumulus cell (CC) expansion and its morphology after in vitro maturation (IVM); (c) to ascertain the influence of in vivo serum progesterone (SP) concentrations of ovary donors on oocyte nuclear maturation, CC phenotypes and degrees of CC expansion of in vitro matured COCs. After 48 h of IVM in modified TCM 199, CCs from grade 1 and 2 COCs were stained with propidium iodide. Oocyte chromatin configuration was visualized by Hoechst 33342 stain. Results showed that oocyte IVM was not influenced by degree of CC expansion (D1, D2 and D3) in COCs. From the CC types (C1, C2 and C3), number of C1 types was higher at D1 expansion and differed from those observed at D2 and D3 expansions. Additionally, rates of apoptosis in D1 CCs were lower than those observed in D2 CCs (p < 0.05). Oocyte nuclear maturation was not influenced by in vivo SP concentrations of ovary donors. On the other hand, D3 expansion prevailed in COCs from bitches at SP > 2.5 ng/ml (p < 0.001). Moreover, in vitro CC apoptosis was associated both with low (0-1 ng/ml) and with high (>5 ng/ml) in vivo SP levels. These findings indicate that morphology of CCs from in vitro matured dog oocytes gives valuable information on viability of COCs and could possibly be used as a parameter in predicting the quality of oocytes destined for in vitro fertilization (IVF) and their outcomes. PMID- 19126266 TI - Erythrocyte invasion profiles are associated with a common invasion ligand polymorphism in Senegalese isolates of Plasmodium falciparum. AB - Plasmodium falciparum parasites use multiple ligand-receptor interactions to invade human erythrocytes. Variant expression levels of members of the PfRh and PfEBA ligand families are associated with the use of different erythrocyte receptors, defining invasion pathways. Here we analyse a major polymorphism, a large sequence deletion in the PfRh2b ligand, and erythrocyte invasion profiles in uncultured Senegalese isolates. Parasites vary considerably in their use of sialic acid-containing and protease-sensitive erythrocyte receptors for invasion. The erythrocyte selectivity index was not related to invasion pathway usage, while parasite multiplication rate was associated with enhanced use of a trypsin resistant invasion pathway. PfRh2b protein was expressed in all parasite isolates, although the PfRh2b deletion was present in a subset (approximately 68%). Parasites with the PfRh2b deletion were found to preferentially utilize protease-resistant pathways for erythrocyte invasion. Sialic acid-independent invasion is reduced in parasites with the PfRh2b deletion, but only in isolates derived from blood group O patients. Our results suggest a significant role for PfRh2b sequence polymorphism in discriminating between alternative erythrocyte receptors for invasion and as a possible determinant of virulence. PMID- 19126267 TI - Mutational, inhibitory and microcalorimetric analyses of Plasmodium falciparum TMP kinase. Implications for drug discovery. AB - Plasmodium falciparum thymidylate kinase (PfTMK) can tolerate a range of substrates, which distinguishes it from other thymidylate kinases. The enzyme not only phosphorylates TMP and dUMP but can also tolerate bulkier purines, namely, dGMP, GMP, and dIMP. In order to probe the flexibility of PfTMK in accommodating ligands of various sizes, we developed 6 mutant enzymes and subjected these to thermodynamic, inhibitory and catalytic evaluation. Kinase activity was markedly affected by introducing a larger lysine residue instead of A111. The lack of the hydroxyl group after inducing mutation of Y107F affected enzyme activity, and had a more severe impact on dGMP kinase activity. PfTMK can be inhibited by both purine and pyrimidine nucleosides, raising the possibility of developing highly selective drugs. Thermodynamic analysis revealed that enthalpic forces govern both purine and pyrimidine nucleoside monophosphate binding, and the binding affinity of both substrates was highly comparable. The heat produced due to dGMP binding is lower than that attributable to TMP. This indicates that additional interactions occur with TMP, which may be lost with larger dGMP. Targeting PfTMK not only affects thymidine nucleotide synthesis but may also affect purine nucleotides, and thus the enzyme represents an attractive antimicrobial target. PMID- 19126268 TI - CrATP as a new inhibitor of ecto-ATPases of trypanosomatids. AB - Trypanosomatid protozoa include heteroxenic species some of them pathogenic for men, animals and plants. Parasite membrane contains ecto-enzymes whose active sites face the external medium rather than the cytoplasm. Herpetomonas sp. displayed a Mg2+-dependent ecto-ATPase activity, a Mg-independent ecto-ADPase and an ecto-phosphatase activity. Both, the ecto-ADPase and phosphatase activities were insensitive to CrATP (chromium(III) adenosine 5'-triphosphate complex). Ecto ATPase activity was reversibly inhibited. At 2 mm ATP the apparent Ki was 4 x 7+/ 1 x 0 microm but a fraction of about 40-50% was insensitive to CrATP. Remarkably, at low substrate concentration (0 x 2 mm) more than 90% of the ecto-ATPase was inhibited with Ki=0 x 33+/-0 x 10 microm. These parameter dependences are interpreted as the presence of 2 ecto-ATPases activities, one of them with high ATP apparent affinity and sensitivity to CrATP. DIDS (4,4 diisothiocyanatostilbene 2,2' disulfonic acid), suramin and ADP were also effective as inhibitors. Only ADP presented no additive inhibition with CrATP. The pattern of partial inhibition by CrATP was also observed for the ecto-ATPase activities of Leishmania amazonensis, Trypanosoma cruzi and Trypanosoma rangeli. CrATP emerges as a new inhibitor of ecto-ATPases and as a tool for a better understanding of properties and role of ecto-ATPases in the biology of parasites. PMID- 19126269 TI - Cysteine peptidases in Herpetomonas samuelpessoai are modulated by temperature and dimethylsulfoxide-triggered differentiation. AB - Cysteine peptidases of protozoa have been implicated in a variety of biological events, and the expression of these enzymes is modulated in response to distinct stimuli, including environmental changes and differentiation. In the present work, we have examined the expression of cysteine peptidases from Herpetomonas samuelpessoai grown at distinct temperatures and during dimethylsulfoxide (DMSO) elicited differentiation. We demonstrated that a 45 kDa cysteine peptidase had its activity reduced during the parasite growth at 37 degrees C in comparison to 26 degrees C, and when cultured up to 72 h in the presence of DMSO. The modulation in the 45 kDa cysteine peptidase expression is connected to the differentiation process, since both temperature and DMSO are able to trigger the promastigote to paramastigote transformation in H. samuelpessoai. The possible immunological similarity of H. samuelpessoai proteins with well-known cysteine peptidases produced by trypanosomatid pathogens, including cruzipain (Trypanosoma cruzi) and cysteine peptidase b (cpb) from Leishmania mexicana, was also investigated, as well as with calpain molecules. The protein cellular lysate of H. samuelpessoai reacted with antibodies raised against cpb of L. mexicana and calpain of Drosophila melanogaster; however, no reaction was observed against cruzipain. The 35 kDa cpb-like protein had its expression diminished in DMSO treated parasites, while the 80 kDa calpain-like molecule was enhanced and an additional 30 kDa calpain-related polypeptide was exclusively observed in these cells. Fluorescence microscopy and flow cytometry analyses corroborated these data. The results described above add H. samuelpessoai to the list of parasites whose differentiation seems to be correlated with cysteine peptidase expression. PMID- 19126270 TI - Flow cytometry analysis of the circulating haemocytes from Biomphalaria glabrata and Biomphalaria tenagophila following Schistosoma mansoni infection. AB - Aiming to further characterize the haemocyte subsets in Biomphalaria snails, we have performed a detailed flow cytometric analysis of whole haemolymph cellular components using a multiparametric dual colour labelling procedure. Ethidium bromide/acridine orange fluorescence features were used to first select viable haemocytes followed by flow cytometric morphometric analysis based on the laser scatter properties (forward scatter-FSC and side scatter-SSC). Our findings demonstrated that B. glabrata (BG-BH, highly susceptible to S. mansoni) and 2 strains of B. tenagophila (BT-CF, moderately susceptible and BT-Taim, resistant to S. mansoni) have 3 major circulating haemocyte subsets, referred to as small, medium and large haemocytes. The frequency of small haemocytes was higher in BG BH, while medium haemocytes were the most abundant cell-type in both B. tenagophila strains. Schistosoma mansoni infection resulted in early reduction of large and medium circulating haemocytes followed by an increase of small haemocytes. Although parasite infection induced haemocyte alterations in all Biomphalaria strains, the response was particularly intense in BT-Taim, the parasite-resistant snail. Interestingly, the trematode infection induces changes in haemocytes with less granular rather than in those with more granular profile. The results indicated that, in B. tenagophila of Taim strain, circulating haemocytes, especially the medium and high subset with less granular profile, are very reactive cells upon S. mansoni infection, suggesting that this cell subset would participate in the early parasite destruction observed in this snail strain. PMID- 19126271 TI - Unique biological rhythm in the reproductive behaviour of female ticks of reptiles. AB - We report the discovery of a biological rhythm in the reproductive behaviour of the tick Bothriocroton hydrosauri that was absent in Amblyomma limbatum, a species that occurs on the same species of reptile host. Female B. hydrosauri mated in autumn or winter delayed oviposition until the following spring, while there was no diapause in conspecific females mated in spring or early summer. Initiation of ovipositional diapause in ticks is usually related to photoperiodic stimuli, but this was not the case for B. hydrosauri. The sinusoidal pattern in pre-oviposition times of B. hydrosauri females mated in different months in the laboratory suggests an internal seasonal time-keeping mechanism. We hypothesize that hormones imbibed by females during their bloodmeal may provide environmental cues associated with the induction of diapause. Irrespective of the mechanism underlying the rhythm, diapause by B. hydrosauri females mated during autumn or winter is of adaptive advantage because it synchronizes oviposition with favourable environmental conditions for egg hatching and increases the chance of larvae finding a host. The lack of a similar biological rhythm in A. limbatum may be a reflection of the different environmental conditions this species experiences throughout most of its range as compared with B. hydrosauri. PMID- 19126272 TI - Morphological diversification in different trematode lineages: body size, host type, or time? AB - Different lineages experience different rates of phenotypic diversification, resulting in greater or lower variance in the expression of phenotypic traits among the species within a lineage. Here, morphological diversification is investigated in 14 different trematode families, based on a dataset comprising morphometric data on body size and 4 anatomical structures (oral sucker, ventral sucker, pharynx, cirrus sac) from 386 species. Three hypotheses are tested and subsequently rejected based on the empirical evidence. First, the degree of morphological variation in all traits within a trematode family, measured as the coefficient of variation among species, appears independent of the average body size of species belonging to that family. Second, patterns of morphological diversification appear similar whether endothermic or ectothermic vertebrates are used as definitive hosts. Third, phylogenetically older trematode lineages did not display greater morphological variation than younger, more derived ones, ruling out evolutionary time as an explanation. The results are consistent with developmental constraints acting on morphological diversification, since for some pairs of traits, variation in one trait is not independent of variation in another trait. More importantly, across most families, variation in body size was significantly more pronounced than variation in the relative sizes of the other morphological features. Trematode body size therefore varies widely while the general body architecture of the family is maintained. The fact that the evolution of the body plan is more conservative than that of body size suggests that the range of morphologies that can evolve in trematodes is constrained. PMID- 19126273 TI - FVB/N mice are highly resistant to primary infection with Nippostrongylus brasiliensis. AB - Nippostrongylus brasiliensis larvae are particularly susceptible to immunological attack during the pre-lung stage of primary and secondary infections in mice. Whilst most of the common laboratory strains of mice are permissive hosts for the parasite, in this study we report for the first time, the strong resistance of naive FVB/N mice to N. brasiliensis. Damage to larvae is evident within the first 24 h of infection and this may be critical to later larval development and reproductive success. Inflammatory responses in the skin, and larval escape from this tissue were comparable in susceptible CBA/Ca and resistant FVB/N mice, with most larvae exiting within 4 h of a primary infection. Lung larval burdens were also similar between strains, but larvae recovered from FVB/N mice were smaller and less motile. In FVB/N mice, larval colonization of the gut was impaired and worms produced very few eggs. However FVB/N mice did not show enhanced resistance to Heligmosomoides bakeri (also known as Heligmosomoides polygyrus), a nematode largely restricted to the gut. Damage done in the pre-lung or lung stages of infection with N. brasiliensis is likely to contribute to ongoing developmental and functional abnormalities, which are profoundly evident in the gut phase of infection. PMID- 19126274 TI - Angiostrongylus vasorum from South America and Europe represent distinct lineages. AB - Angiostrongylus vasorum is a nematode parasite of sylvan and domestic species of the family Canidae. It has a broad but patchy distribution worldwide, and there is evidence for geographical spread and increasing incidence of infection in recent years. While historically Angiostrongylus-like nematodes identified in dogs and foxes have been described as A. vasorum in Europe and Angiocaulus raillieti in South America, more recent taxonomic revision has amalgamated these into a single species, A. vasorum. Here we report, for the first time, the molecular characterization of isolates of A. vasorum from Germany, Portugal, Denmark and the United Kingdom on the basis of the mitochondrial COI gene and the second ribosomal internal transcribed spacer. When compared with isolates from Brazil, sequence analysis revealed 2 distinct genotypes. Estimated rates of evolution based on COI sequences for both nematode and host are consistent with the hypothesis that the presence of A. vasorum in South America is a result of an ancient evolutionary event. Angiostrongylus vasorum in South America potentially represents a separate species to that observed in Europe. PMID- 19126275 TI - Effects of host characteristics and parasite intensity on growth and fecundity of Trichostrongylus retortaeformis infections in rabbits. AB - Host-mediated responses and parasite density-dependent processes can have a major influence on the growth and fecundity of parasitic nematodes. However, host characteristics and parasite intensity consistently change during the course of an infection and these could affect worm length and number of eggs in a non constant way. We used a free-living population of rabbits naturally infected with Trichostronglyus retortateformis and examined how adult nematode length and the number of eggs in utero were affected by host characteristics (i.e. age, sex, breeding status) and parasite intensity, in a seasonal environment, between 2004 and 2007. Nematode length and number of eggs in utero decreased exponentially with host age; in contrast, parasite intensity increased, peaked in juveniles and decreased in older hosts. These patterns were consistent between rabbit cohorts. A negative relationship was observed between parasite intensity and nematode length, as well as number of eggs. Nematode length was strongly affected by nematode sex and host age, while the number of eggs was mainly influenced by nematode length. The direct influence of host-mediated effects appeared quantitatively more important than parasite density dependence in controlling length and egg production in naturally infected wild rabbits. However, their relative contribution changed during the course of the infection such that, while host immunity still influenced worm numbers, the direct effect of density dependent interactions contributed the most at high parasite intensities. PMID- 19126276 TI - A fatal true knot of cord. PMID- 19126278 TI - Therapeutic controversies. PMID- 19126280 TI - Diagnosis, management, and evaluation of hypertensive disorders of pregnancy. PMID- 19126281 TI - Blood transfusion for primary postpartum hemorrhage: a tertiary care hospital review. AB - OBJECTIVE: To describe the common characteristics, clinical management, and outcome of patients requiring blood transfusion within 24 hours of delivery. METHODS: We conducted a retrospective cohort study of patients who received blood transfusion for postpartum hemorrhage (PPH) in the first 24 hours post-delivery, over a five-year period (2000-2005). The medical records of patients were reviewed to obtain information about demographics, pregnancy and delivery characteristics, transfusion data, and complications. RESULTS: The overall blood transfusion rate for PPH was 0.31% (104/33,631 deliveries). The rate of blood transfusion in women who had a Caesarean section during labour was 0.49%, whereas in women who had a vaginal delivery or elective Caesarean section it was 0.28% and 0.23%, respectively. Antenatal risk factors for PPH were identified in 61% of patients, and 39% of patients developed intrapartum risk factors. The most important etiological factors were uterine atony (38.5%) and retained products of conception (33.7%). Twenty-one percent of the patients developed coagulopathy, and 24% required admission to the intensive care unit. CONCLUSION: Severe primary PPH requiring blood transfusion can be predicted in the majority of patients on the basis of antenatal risk factors, while the remaining patients require vigilant monitoring for risk factors during labour and delivery. In the multidisciplinary effort to prevent and control major PPH, we should re-evaluate the pharmacotherapy for PPH and ensure careful removal of retained placental tissue after delivery. PMID- 19126282 TI - Intrahepatic cholestasis of pregnancy in women with a multiple pregnancy: an analysis of risks and pregnancy outcomes. AB - OBJECTIVE: This study was conducted to assess the incidence and perinatal outcomes of multiple pregnancies complicated by intrahepatic cholestasis of pregnancy in an urban population. METHODS: We performed a retrospective chart review of all multiple gestation deliveries at our institution between January 2004 and December 2005. Antepartum and delivery data were collected for all patients. Symptoms and treatment were also abstracted for patients in whom intrahepatic cholestasis was diagnosed. We used the Student two-tail t test and Fisher exact test to examine the differences between multiple gestation pregnancies with and without cholestasis of pregnancy. RESULTS: Data were available for 263 multiple pregnancies. The incidence of cholestasis was 4.2% (11/263), with a mean onset at 29.4 weeks. There were no differences in mean gestational age at delivery, preterm delivery rate, meconium histiocytosis, incidence of preeclampsia, or incidence of postpartum hemorrhage between women with and those without cholestasis. There were no intrauterine fetal deaths in the cholestasis group. CONCLUSION: Women with multiple gestations complicated by cholestasis of pregnancy do not have increased adverse perinatal outcomes. The absence of unexplained fetal demise may be a result of routine delivery before 40 weeks' gestation in multiple pregnancies. PMID- 19126283 TI - SOGC Partnership Program 1998-2006: building organization capacity to support improved maternal and neonatal health. AB - Through its partnership program with professional obstetrics and gynaecology associations in resource-poor environments, the Society of Obstetricians and Gynaecologists of Canada has developed a model for capacity assessment and development. The use of this model in Guatemala, Haiti, and Uganda has demonstrated its potential for facilitating cultural shifts in the perception of reproductive care, for providing clinical expertise in promoting educational activities, for enabling enhanced technical capacity, for political engagement, and for broad-based community and professional engagement. PMID- 19126284 TI - Human papillomavirus--lessons from history and challenges for the future. AB - Human papillomavirus (HPV) is the most common sexually transmitted infection, and HPV-associated cervical cancer is a significant cause of morbidity and mortality worldwide. Recent advances in molecular biology have facilitated testing for HPV infection. Over the last decade, national and international cervical cancer screening programs have added HPV testing to their guidelines. The use of HPV prophylactic and therapeutic immunization may expand the need for systematic HPV testing to help define eligible subgroups for intervention. Given the worldwide variation in HPV subtype prevalence, basic Pap testing will continue to play an important role in cervical cancer screening, and methods to improve Pap smear sensitivity may help to improve screening in the future. This review focuses on the genetics and cellular biology of HPV infection, the natural history and prevalence of HPV infections, cervical cancer screening around the world and in Canada in particular, and evolving research to improve screening methods. PMID- 19126285 TI - Suprapubic or urethral catheter: what is the optimal method of bladder drainage after radical hysterectomy? AB - BACKGROUND: Lower urinary tract dysfunction is a common morbidity related to radical hysterectomy (RAH). Although transurethral catheterization (TUC) has traditionally been used for postoperative bladder drainage following RAH, suprapubic catheterization (SPC) is an alternative method that may be advantageous. OBJECTIVES: To determine, by means of a retrospective cohort study, the incidence of urinary tract infection (UTI), duration of postoperative hospital stay, and time to trial of voiding in women catheterized suprapubically or transurethrally after RAH for early stage cervical cancer. METHODS: Two hundred twelve patients who underwent RAH and staging for stage IA1 + LVS, 1A2, and 1B1 cancer of the cervix in Edmonton between 1996 and 2006 were included in the study. Three gynaecologic oncologists performed the surgeries. Operative, postoperative, and demographic data were extracted from patient records. Patients were catheterized either suprapubically (SPC group) or transurethrally (TUC group) according to the surgeon's discretion. Comparative tests and multivariate regression analysis were used to compare outcome measures between the groups and to adjust for confounding variables. RESULTS: The TUC group had a higher proportion of patients with UTI (27%) than the SPC group (6%) (P < 0.001). The SPC group had a shorter postoperative hospital stay (4.8 vs. 5.7 days; P < 0.001) and an earlier trial of voiding (2.7 vs. 4.4 days; P < 0. 001). Following regression analysis, statistically significant differences remained for UTI and time to initiation of a trial of voiding. CONCLUSION: After RAH for early stage cervical cancer, suprapubic catheterization is associated with a lower rate of UTI and an earlier trial of voiding than transurethral catheterization. PMID- 19126286 TI - Total laparoscopic versus vaginal hysterectomy: a retrospective comparison. AB - OBJECTIVES: To compare the outcomes of total laparoscopic hysterectomy (TLH) and vaginal hysterectomy (VH) in a community teaching hospital. METHODS: A retrospective chart review of all women who underwent TLH and VH at North York General Hospital between January 2002 and December 2004 was conducted. Groups were compared with respect to demographic data, and intraoperative and postoperative outcomes. RESULTS: One hundred nine women underwent TLH, and 43 women underwent VH. There was no statistically significant difference between groups in the mean age of patients, estimated blood loss, or mean postoperative hemoglobin change. TLH took significantly longer to perform (104.4 vs. 54.4 minutes, P < 0.001); however, the mean postoperative duration of hospital stay was greater in the VH group (2.2 vs. 1.2 days, P < 0.001). Mean uterine mass was significantly greater in the TLH group (290.9 vs. 151.6 g, P < 0. 001). The incidence of intraoperative and postoperative complications was slightly higher in the TLH group than in the VH group, but the difference did not reach statistical significance. CONCLUSIONS: TLH was associated with significantly longer operative time and shorter hospital stay than VH. There was a trend towards more intraoperative and postoperative complications in the TLH group than in the VH group. PMID- 19126287 TI - Diagnostic and operative laparoscopy in Alberta 1994-2006. AB - OBJECTIVES: Laparoscopic surgery has become an important procedure in the diagnosis and management of pelvic pathology. Whether or not repeat laparoscopic procedures, sometimes repeated several times, are of real benefit is unclear. This study was designed to quantify the experience of one province during the period 1994-2006 to explore patterns of laparoscopic surgery. METHODS: Data were obtained from Alberta Health and Wellness regarding diagnostic and operative laparoscopic procedures for the years 1994-2006. The postal code of each responsible health facility was recorded and information grouped into the current nine health regions. The use of laparoscopy in individual women was analyzed by aggregating data by region on an annual basis and then across the entire period of the study. RESULTS: During the years 1994-2006, 51,790 diagnostic laparoscopy procedures and 24,896 operative laparoscopic procedures were performed. Repeat operative rates for 24,473 women were limited when annual rates were analyzed (< 2.0%), but became substantial across the period of study, during which time 36% of women had two or more, 12% had three or more, 5% had four or more, and 2% had five or more operative procedures. CONCLUSIONS: Over an extended period of time, there has been a decline in the rate of diagnostic laparoscopy but not of operative laparoscopy, and there has been a substantial rate of repeat operative laparoscopy per individual woman. These data suggest there may be important reasons for the rates of repeat operative laparoscopy that warrant exploration and the development of practice guidelines. PMID- 19126290 TI - The journal of pharmacy and pharmacology 2008. PMID- 19126288 TI - Missed hormonal contraceptives: new recommendations. AB - OBJECTIVE: To provide evidence-based guidance for women and their health care providers on the management of missed or delayed hormonal contraceptive doses in order to prevent unintended pregnancy. EVIDENCE: Medline, PubMed, and the Cochrane Database were searched for articles published in English, from 1974 to 2007, about hormonal contraceptive methods that are available in Canada and that may be missed or delayed. Relevant publications and position papers from appropriate reproductive health and family planning organizations were also reviewed. The quality of evidence is rated using the criteria developed by the Canadian Task Force on Preventive Health Care. BENEFITS, HARMS, AND COSTS: This committee opinion will help health care providers offer clear information to women who have not been adherent in using hormonal contraception with the purpose of preventing unintended pregnancy. SPONSORS: The Society of Obstetricians and Gynaecologists of Canada. SUMMARY STATEMENTS: 1. Instructions for what women should do when they miss hormonal contraception have been complex and women do not understand them correctly. (I) 2. The highest risk of ovulation occurs when the hormone-free interval is prolonged for more than seven days, either by delaying the start of combined hormonal contraceptives or by missing active hormone doses during the first or third weeks of combined oral contraceptives. (II) Ovulation rarely occurs after seven consecutive days of combined oral contraceptive use. (II) RECOMMENDATIONS: 1. Health care providers should give clear, simple instructions, both written and oral, on missed hormonal contraceptive pills as part of contraceptive counselling. (III-A) 2. Health care providers should provide women with telephone/electronic resources for reference in the event of missed or delayed hormonal contraceptives. (III-A) 3. In order to avoid an increased risk of unintended pregnancy, the hormone-free interval should not exceed seven days in combined hormonal contraceptive users. (II-A) 4. Back-up contraception should be used after one missed dose in the first week of hormones until seven consecutive days of correct hormone use are established. In the case of missed combined hormonal contraceptives in the second or third week of hormones, the hormone-free interval should be eliminated for that cycle. (III-A) 5. Emergency contraception and back-up contraception may be required in some instances of missed hormonal contraceptives, in particular when the hormone-free interval has been extended for more than seven days. (III-A) 6. Back-up contraception should be used when three or more consecutive doses/days of combined hormonal contraceptives are missed in the second and third week until seven consecutive days of correct hormone use are established. For practical reasons, the scheduled hormone-free interval should be eliminated in these cases. (II-A) 7. Emergency contraception is rarely indicated for missed combined hormonal contraceptives in the second or third week of the cycle unless there are repeated omissions or failure to institute back-up contraception after the missed doses. In cases of repeated omissions of combined hormonal contraceptives, emergency contraception may be required, and back-up contraception should be used. Health care professionals should counsel women in these situations on alternative methods of contraception that do not demand such stringent compliance. (III-A). PMID- 19126291 TI - Cancer, chitosan nanoparticles and catalytic nucleic acids. AB - OBJECTIVES: The aim of this review was to examine gene therapy involving DNAzyme and siRNA encapsulation into chitosan nanoparticles, discussing the current and future status of this drug delivery system in enhancing drug delivery and cancer therapy. KEY FINDINGS: Cancer is a disease state in which the cells in our body undergo mutations at the genetic level and are transformed, acquiring the ability to replicate limitlessly. Conventional cancer treatment involves the use of surgery and cytotoxic chemotherapy and/or radiotherapy, which have the potential of harming normal, otherwise healthy, non-neoplastic cells. Newer forms of therapy such as immunotherapy and gene therapy have shown initial promise, but still require better ways to limit exposure to cancerous lesions in the body. As a result drug delivery systems have been developed in attempts to deliver therapeutics specifically to the target lesion site. One recent drug delivery system has revolved around the use of chitosan nanoparticle technology, where therapeutics are encapsulated into nanoparticles and targeted to tumours. SUMMARY: Though few, attempts at encapsulating therapeutics such as deoxyribozymes and small or short interfering RNA have been optimistic and encouraging. PMID- 19126292 TI - Curcuma zedoaria Rosc. (white turmeric): a review of its chemical, pharmacological and ethnomedicinal properties. AB - OBJECTIVES: Curcuma zedoaria Rosc is a perennial herb found in tropical countries, such as India, Japan and Thailand. Various parts of this plant are used in Ayurveda and other folk medicines for the treatment of different ailments such as diarrhoea, cancer, flatulence and dyspepsia. This study is an attempt to compile an up-to-date and comprehensive review of C. zedoaria that covers its traditional and folk medicinal uses, phytochemistry and pharmacology. KEY FINDINGS: Research carried out using different in-vitro and in-vivo techniques of biological evaluation supports most of the claims. SUMMARY: This review presents the botany, chemistry, traditional uses and pharmacological data of the plant. PMID- 19126293 TI - Improving glyburide solubility and dissolution by complexation with hydroxybutenyl-beta-cyclodextrin. AB - OBJECTIVES: Glyburide, an important drug for type 2 diabetes, has extremely poor aqueous solubility and resulting low bioavailability. This study describes the ability of hydroxybutenyl-beta-cyclodextrin (HBenBCD) to form complexes with glyburide, with enhanced solubility and dissolution rate in vitro. METHOD: Glyburide and glyburide-HBenBCD were evaluated in various test media known to simulate human gastrointestinal conditions in the fasted and fed states, respectively. KEY FINDINGS: At approximately 14 wt% drug load, in the presence of HBenBCD, an almost 400-fold increase in glyburide aqueous solubility was observed. In the presence of HBenBCD, glyburide solubility was also significantly improved in all physiologically relevant test media. Subsequent dissolution experiments confirmed the solubility study results; the dissolution rate and total amount of drug released were significantly increased. CONCLUSIONS: Complexation with HBenBCD may be an effective way to increase the bioavailability of glyburide. PMID- 19126294 TI - The role of molecular physicochemical properties and apolipoproteins in association of drugs with triglyceride-rich lipoproteins: in-silico prediction of uptake by chylomicrons. AB - OBJECTIVES: The uptake of drugs by chylomicrons is a key element in both intestinal lymphatic transport and postprandial alterations in the disposition profile of lipophilic drugs. The aim of this article was to elucidate the factors that affect this phenomenon. METHODS: The degree of association of 22 model lipophilic molecules with rat chylomicrons was assessed and correlated in silico with calculated physicochemical properties. The in-silico model was then validated using an external set of molecules. The uptake by chylomicrons was also compared to the association with a marketed artificial emulsion. KEY FINDINGS: The most important physicochemical property that affects the affinity to chylomicrons was found to be LogD7.4; however, a multiparameter model was required to describe properly the uptake process. The in-silico model (R2Y=0.91, R2X=0.91 and Q2=0.82) that was created using a combination of eight molecular descriptors enabled successful prediction of the affinity of the external set of molecules to chylomicrons. The association with the artificial emulsion was statistically different from the uptake by chylomicrons for four (out of nine) molecules. CONCLUSIONS: The association of drugs with chylomicrons is a complex process, which involves the lipophilic core as well as surface apoproteins. The in-silico model based on multiple physicochemical properties of the drugs is able to predict successfully the degree of association with chylomicrons. PMID- 19126295 TI - Pectin-based microspheres for colon-specific delivery of vancomycin. AB - OBJECTIVES: The aim of this study was to describe a colon-specific delivery system based on pectin hydrogels formed by complexation with chitosan. METHODS: Hydrogels were prepared at different weight ratios (4:1, 7:1, 10:1; pectin/chitosan), loaded with vancomycin hydrochloride (2:1, 4:1; polymer/drug weight ratio) and collected by spray-drying. The microspheres obtained were characterized in terms of morphology, swelling behaviour, mucoadhesive properties and drug loading efficiency. The influence of different pectin/chitosan hydrogels on the release behaviour of microspheres at pH 2.0, 5.5 and 7.4 were evaluated in vitro with and without pectinolytic enzyme. KEY FINDINGS: The results showed that water uptake was increased by raising the environmental pH (from 2.0 to 7.4) and the pectin/chitosan weight ratio, while drug availability was increased by raising the environmental pH (from 2.0 to 7.4) and decreased by raising the pectin/chitosan weight ratio. In the presence of pectinase, the glycoside bonds of pectin were degraded and a considerable amount of drug was released in a short time. CONCLUSIONS: This study suggested that pectin/chitosan microspheres were able to limit the release of vancomycin under acidic conditions and release it under simulated colonic conditions, confirming their potential for a colon specific drug delivery system. PMID- 19126296 TI - Effects of cytochrome P450 inducers and inhibitors on the pharmacokinetics of intravenous furosemide in rats: involvement of CYP2C11, 2E1, 3A1 and 3A2 in furosemide metabolism. AB - OBJECTIVES: It has been reported that the non-renal clearance of furosemide was significantly faster in rats pretreated with phenobarbital but was not altered in rats pretreated with 3-methylcholanthrene. However, no studies on other cytochrome P450 (CYP) isozymes have yet been reported in rats. METHOD: Furosemide 20 mg/kg was administered intravenously to rats pretreated with various CYP inducers--3-methylcholanthrene, orphenadrine citrate and isoniazid, inducers of CYP1A1/2, 2B1/2 and 2E1, respectively, in rats--and inhibitors--SKF-525A (a non specific inhibitor of CYP isozymes), sulfaphenazole, cimetidine, quinine hydrochloride and troleandomycin, inhibitors of CYP2C6, 2C11, 2D and 3A1/2, respectively, in rats. KEY FINDINGS: The non-renal clearance of furosemide was significantly faster (55.9% increase) in rats pretreated with isoniazid, but slower in those pretreated with cimetidine or troleandomycin (38.5% and 22.7% decreases, respectively), than controls. After incubation of furosemide with baculovirus-infected insect cells expressing CYP2C11, 2E1, 3A1 or 3A2, furosemide was metabolized via CYP2C11, 2E1, 3A1 and 3A2. CONCLUSIONS: These findings could help explain possible pharmacokinetic changes of furosemide in various rat disease models (where CYP2C11, 2E1, 3A1 and/or CYP3A2 are altered) and drug-drug interactions between furosemide and other drugs (mainly metabolized via CYP2C11, 2E1, 3A1 and/or 3A2). PMID- 19126297 TI - Concentration-dependent plasma protein binding of the novel dipeptidyl peptidase 4 inhibitor BI 1356 due to saturable binding to its target in plasma of mice, rats and humans. AB - OBJECTIVES: The purpose of this study was to characterise the plasma protein binding of BI 1356. METHODS: BI 1356 (proposed trade name ONDERO) is a novel dipeptidyl peptidase 4 (DPP-4) inhibitor, which is under clinical development for the treatment of type 2 diabetes. DPP-4 is expressed in various tissues but soluble DPP-4 is also present in plasma. Therefore, binding to soluble DPP-4 may influence the pharmacokinetics of BI 1356. Plasma protein binding of BI 1356 was determined in vitro for wild type mice and rats and the results compared with those for DPP-4 knockout mice and DPP-4 deficient Fischer rats. In addition, protein binding of BI 1356 was examined in plasma from healthy human volunteers and renal excretion of the compound in the DPP-4 knockout mice was compared with that occurring in wild type mice. KEY FINDINGS: The results showed that BI 1356 exhibited a prominent concentration-dependent plasma protein binding due to a saturable high affinity binding to the DPP-4 target in plasma. Differences in renal excretion of BI 1356 between DPP-4 knockout mice and wild type mice suggested that saturable binding of BI 1356 to DPP-4 in the body also influenced elimination. CONCLUSIONS: High affinity, but readily saturable binding of BI 1356 to its target DPP-4 accounted primarily for the concentration-dependent plasma protein binding at therapeutic plasma concentrations of BI 1356. PMID- 19126298 TI - Human phenylalanine monooxygenase and thioether metabolism. AB - OBJECTIVES: The substrate specificity of wild-type human phenylalanine monooxygenase (wt-hPAH) has been investigated with respect to the mucoactive drug, S-carboxymethyl-L-cysteine and its thioether metabolites. The ability of wt hPAH to metabolise other S-substituted cysteines was also examined. METHODS: Direct assays of PAH activity were by HPLC with fluorescence detection; indirect assays involved following disappearance of the cofactor by UV spectroscopy. KEY FINDINGS: wt-hPAH catalysed the S-oxygenation of S-carboxymethyl-L-cysteine, its decarboxylated metabolite, S-methyl-L-cysteine, and both their corresponding N acetylated forms. However, thiodiglycolic acid was not a substrate. The enzyme profiles for both phenylalanine and S-carboxymethyl-L-cysteine showed allosteric kinetics at low substrate concentrations, with Hill constants of 2.0 and 1.9, respectively, for the substrate-activated wt-hPAH. At higher concentrations, both compounds followed Michaelis-Menten kinetics, with non-competitive substrate inhibition profiles. The thioether compounds, S-ethyl-L-cysteine, S-propyl-L cysteine and S-butyl-L-cysteine were all found to be substrates for phenylalanine monooxygenase. CONCLUSIONS: Phenylalanine monooxygenase may play a wider role outside intermediary metabolism in the biotransformation of dietary-derived substituted cysteines and other exogenous thioether compounds. PMID- 19126299 TI - Andrographolide inhibits growth of acute promyelocytic leukaemia cells by inducing retinoic acid receptor-independent cell differentiation and apoptosis. AB - OBJECTIVES: The growth inhibiting potential of andrographolide was evaluated in three acute promyelocytic leukaemia cell line models (HL-60, NB4 and all-trans retinoic acid (ATRA)-resistant NB4-R2). METHODS: In elucidating the mechanisms of growth inhibition, a special emphasis was placed on assessing the induction of differentiation and apoptosis by andrographolide in the primary acute promyelocytic leukaemia NB4 cells. KEY FINDINGS: The compound was 2- and 3-fold more active in inhibiting the growth of HL-60 and NB4-R2 cells compared with NB4 cells, respectively. At IC50 (concentration at which growth of 50% of the cells (compared with medium only treated control cells) is inhibited; 4.5 microM) the compound exhibited strong cell-differentiating activity in NB4 cells, similar to ATRA (IC50 1.5 microM). In the presence of a pure retinoic acid receptor antagonist AGN193109, the growth inhibition of NB4 cells by ATRA was reversed, whereas the activity of andrographolide was not affected. This clearly suggested that andrographolide's cell differentiating activity to induce growth inhibition of NB4 cells most likely occurred via a retinoic acid receptor-independent pathway. At higher concentration (2xIC50), andrographolide was an efficient inducer of apoptosis in NB4 cells. CONCLUSIONS: Taken together, these results suggest andrographolide and its derivatives, apparently with a novel cell differentiating mechanism and with ability to induce apoptosis, might be beneficial in the treatment of primary and ATRA-resistant acute promyelocytic leukaemia. PMID- 19126300 TI - Increased sensitivity to diltiazem hypotensive effect in an experimental model of high-renin hypertension. AB - OBJECTIVES: The aim of this work was to evaluate the pharmacokinetic pharmacodynamic properties of diltiazem in an experimental model of high-renin hypertension, such as the aortic coarctated (ACo) rat, to further characterize the responsiveness of this model to calcium channel blockers. METHODS: A 'shunt' microdialysis probe was inserted in a carotid artery of anaesthetized ACo and control sham-operated (SO) rats for simultaneous determination of diltiazem plasma concentrations and their effects on mean arterial pressure and heart rate after the intravenous application of 3 and 6 mg/kg of the drug. Correlation between plasma levels and cardiovascular effects was established by fitting the data to a modified Emax model. KEY FINDINGS: Volume of distribution was greater in ACo than in SO rats. Diltiazem plasma clearance (Cl) was significantly greater in ACo rats than in normotensive SO rats after administration of diltiazem (6 mg/kg). Moreover, Cl increased with dose in ACo but not in SO rats. No differences were observed in the maximal bradycardic effect comparing both experimental groups, and sensitivity (S0) to diltiazem chronotropic effect was similar comparing SO and ACo rats. Differences were not found in the maximal response of the hypotensive effect comparing SO and ACo rats, but the S0 to diltiazem hypotensive effect was greater in ACo rats than in SO rats. CONCLUSIONS: ACo induced profound changes in diltiazem pharmacokinetic behaviour. In addition, our results suggested an increased sensitivity to diltiazem blood pressure lowering effect in experimental renovascular hypertension with high renin levels. PMID- 19126301 TI - Anticoagulant and antithrombotic activity of a new peptide pENW (pGlu-Asn-Trp). AB - OBJECTIVES: The aim was to test a newly discovered oligopeptide, pENW (pGlu-Asn Trp), for its anticoagulant and antithrombotic activity in vivo, and try to investigate its underlying mechanisms. METHODS: We measured coagulation time by the glass slide method and bleeding time by cutting of mice tails. The thrombosis models employed here included an arterio-venous shunt model and inferior vena ligation model. An ELISA (enzyme-linked immunosorbent assay) was used to analyse t-PA/PAI (tissue-type plasminogen activator/plasminogen activator inhibitor) in the blood drawn from the rats with thrombosis. The ultrastructural changes of the endothelium in the vessels developing thrombosis were observed under a transmission electron microscope. KEY FINDINGS: We found that pENW-treated mice exhibited a prolonged coagulation time in a dose-dependent manner, but not an extended haemorrhage time. On the other hand, pENW significantly inhibited thrombus formation in both arterio-venous shunt models and inferior vena ligation models. Plasma t-PA/PAI was significantly higher as measured by ELISA. Transmission electron microscope photos of pENW-treated groups also displayed a better condition than model controls, with less erythrocytes in the vascular lumens. In addition, pENW concentration-dependently inhibited aggregation of platelets induced by ADP (adenosine 5'-diphosphate sodium salt) in rabbit platelet-rich plasma. CONCLUSIONS: These findings support the suggestion that pENW possesses antithrombotic activity and could be a promising drug in the prevention and treatment of unwanted clot formation. PMID- 19126302 TI - Effect of traditional Chinese herbal Bu-Wang-San on synaptic plasticity in ovariectomised rats. AB - OBJECTIVES: The neuroprotective effects of Bu-Wang-San (BWS) and its effects on spine synapse plasticity were investigated in ovariectomised rats. METHODS: Thirty-six ovariectomised rats were divided into three groups: untreated controls, treatment with 17beta-estradiol or with BWS. After 3 months, spatial acquisition and spatial retention were measured using the Morris water maze. Swim time, swim distance, swim speed, quadrant time and platform crossing were recorded. Spine synapse density in the hippocampus was examined by transmission electron microscopy. The expression of synaptophysin P38 (P38) mRNA was examined by real-time PCR and the protein expression of P38 was examined by Western blot. KEY FINDINGS: In spatial acquisition and spatial retention, the BWS group functioned significantly better than the control group. Ultrastructural observation of the hippocampus showed that BWS significantly increased spine synapse density compared with the ovariectomised group. In addition, BWS significantly increased P38 mRNA and protein expression in the hippocampus. Thus, the positive effect of BWS on learning and memory in rats was associated with increased spinal synapse density and increased P38 mRNA and protein expression in the hippocampus following menopause-induced injury. CONCLUSIONS: These results suggest that BWS could improve cognitive ability following menopause-induced impairment of learning and memory. PMID- 19126303 TI - Triclisia sacleuxii (Pierre) Diels (Menispermaceae), a potential source of acetylcholinesterase inhibitors. AB - OBJECTIVES: To search for compounds possibly useful for the treatment of Alzheimer's disease. METHODS: Alkaloid fractions from the roots, stems and leaves of Triclisia sacleuxii (Menispermaceae) and pure bisbenzylisoquinoline alkaloids isolated from the roots (phaeanthine, N-methylapateline, 1,2-dehydroapateline and gasabiimine) were assessed for acetylcholinesterase inhibitory activity. KEY FINDINGS: All extracts and compounds tested inhibited acetylcholinesterase to varying degrees; the leaf tertiary alkaloid fractions and the root quaternary alkaloid fractions exhibited the strongest inhibitory potential (90% at 0.1 mg/ml). The leaf tertiary alkaloid fraction was selected for further analysis (the quaternary alkaloids, which are too polar for absorption and brain distribution, were presumed to be clinically uninteresting). TLC bioautography using Ellman's reagent allowed the detection of acetylcholinesterase inhibitors and the isolation of the major active constituent, which was identified as lindoldhamine, a one-bridged bisbenzylisoquinoline alkaloid. Lindoldhamine displayed high acetylcholinesterase inhibitory activity with a 50% inhibition concentration in the micromolar range. CONCLUSIONS: All T. sacleuxii alkaloid fractions tested exhibited anti-acetylcholinesterase activity; isolated bisbenzylisoquinoline alkaloids showed weak-to-high inhibition depending on their structural features. Structure modification could provide interesting derivatives with enhanced anti-acetylcholinesterase activity. PMID- 19126304 TI - Effect of natsudaidain isolated from Citrus plants on TNF-alpha and cyclooxygenase-2 expression in RBL-2H3 cells. AB - OBJECTIVES: Flavonoids inhibit the activity of chemical mediators released from mast cells. Our aim was to investigate the effects of natsudaidain, a polymethoxyflavone isolated from Citrus plants, on mast cells. METHODS: We investigated the inhibitory effects of natsudaidain, which is a polymethoxyflavone isolated from Citrus plants, on histamine release, tumour necrosis factor-alpha production and cyclooxygenase-2 expression in Ca ionophore stimulated rat basophilic leukemia cells (A23187-stimulated RBL-2H3 cells) by spectrofluorometric, ELISA and immunoblotting methods. KEY FINDINGS: The percent of histamine release from A23187-stimulated RBL-2H3 cells pretreated with natsudaidain at 5, 25 and 50 microM was not changed as compared with non-treated A23187-stimulated cells. At 100 and 200 microM, natsudaidain pretreatment resulted in slightly reduced histamine release (% histamine release, 89.8+/-3.5% and 71.5+/-5.6% at 100 and 200 microM). Thus, natsudaidain hardly affects histamine release from RBL-2H3 cells, except at high concentrations. On the other hand, natsudaidain dose-dependently inhibited tumour necrosis factor-alpha protein and mRNA levels in A23187-stimulated RBL-2H3 cells; a concentration of 6.8 microM was required for a 50% reduction. In addition, all concentrations of this compound that we tested also inhibited cyclooxygenase-2 protein expression. The mRNA levels of cyclooxygenase-2 in A23187-stimulated RBL-2H3 cells treated with natsudaidain were also markedly decreased. The phosphorylated-p38 MAPK protein levels in A23187-stimulated RBL-2H3 cells treated with natsudaidain were lower than in the non-treated cells. CONCLUSIONS: These findings suggest that natsudaidain inhibits tumour necrosis factor-alpha and cyclooxygenase-2 production by suppressing p38 MAPK phosphorylation but not p65 NFkappaB phosphorylation, and that natsudaidain might alleviate inflammatory diseases. PMID- 19126305 TI - Inhibitory effect of quercetin on rat trachea contractility in vitro. AB - OBJECTIVES: The effect of quercetin, a naturally occurring flavonoid traditionally used to treat airway diseases such as bronchial asthma, on the contractile response elicited by electrical field stimulation or carbachol in rat isolated trachea was investigated. METHODS: Isolated tracheal tissue was subjected to contractions by an electrical field stimulation of 5 Hz for 30 s, 400 mA, and the responses in the presence of cumulative concentrations of quercetin (10(-6)-3x10(-4) M) were observed. The effect of quercetin was also evaluated after administration of phentolamine plus propranolol (to block alpha- and beta-adrenergic receptors), NG-nitro-L-arginine methyl ester (to block nitric oxide synthesis), capsaicin (to desensitise sensory C fibres), alpha-chymotrypsin (a proteolytic enzyme that rapidly degrades vasoactive intestinal peptide), SR140333 and SR48968 (tackykinin NK1 and NK2 receptor antagonists, respectively). KEY FINDINGS: Quercetin produced a concentration-dependent inhibition of contractions induced by both carbachol and electrical field stimulation. However, quercetin was more active in inhibiting the contractions produced by electrical field stimulation than those induced by carbachol, suggesting a presynaptic site of action (in addition to a postsynaptic effect, as revealed by the inhibitory action of quercetin on carbachol-induced contractions). The inhibitory effect of quercetin on contractions induced by electrical field stimulation was unaffected by phentolamine plus propranolol, SR 140333 and SR 48968, capsaicin treatment or by the proteolytic enzyme alpha-chymotrypsin. In contrast, the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester significantly reduced the inhibitory effect of quercetin on contractions induced by electrical field stimulation. CONCLUSIONS: Quercetin inhibits rat tracheal contractility through a presynaptic (involving nitric oxide) and a postsynaptic site of action. PMID- 19126306 TI - Fish skin as a model membrane: structure and characteristics. AB - OBJECTIVES: Synthetic and cell-based membranes are frequently used during drug formulation development for the assessment of drug availability. However, most of the currently used membranes do not mimic mucosal membranes well, especially the aqueous mucous layer of the membranes. In this study we evaluated catfish (Anarichas lupus L) skin as a model membrane. METHOD: Permeation of hydrocortisone, lidocaine hydrochloride, benzocaine, diethylstilbestrol, naproxen, picric acid and sodium nitrate through skin from a freshly caught catfish was determined in Franz diffusion cells. KEY FINDINGS: Both lipophilic and hydrophilic molecules permeate through catfish skin via hydrated channels or aqueous pores. No correlation was observed between the octanol/water partition coefficient of the permeating molecules and their permeability coefficient through the skin. Permeation through catfish skin was found to be diffusion controlled. CONCLUSIONS: The results suggest that permeation through the fish skin proceeds via a diffusion-controlled process, a process that is similar to drug permeation through the aqueous mucous layer of a mucosal membrane. In addition, the fish skin, with its collagen matrix structure, appears to possess similar properties to the eye sclera. PMID- 19126307 TI - Pharmacokinetic comparison of puerarin after oral administration of Jiawei Xiaoyao-San to healthy volunteers and patients with functional dyspepsia: influence of disease state. AB - OBJECTIVES: An investigation has been designed and conducted to compare the pharmacokinetics of puerarin after oral administration of Jiawei-Xiaoyao-San to healthy volunteers and to patients with functional dyspepsia. METHODS: Quantification of puerarin in serum was achieved using a simple and rapid HPLC method for pharmacokinetic study. KEY FINDINGS: After oral administration of decoctions of Jiawei-Xiaoyao-San to healthy volunteers and patients with functional dyspepsia, puerarin was absorbed and reached a maximum concentration of 56.47+/-15.60 and 29.40+/-8.89 ng/ml at 51.00+/-8.22 and 50.00+/-12.25 min, respectively. Compared with the value of AUC0-360 (5.55+/-1.47 microg/ml min) in healthy volunteers, a smaller value of AUC0-360 (3.35+/-1.17 microg/ml min) was observed in patients with functional dyspepsia. CONCLUSIONS: There were statistically significant differences in the pharmacokinetic parameters of puerarin including the values for Cmax, AUC0-360, Cl/F and MRT0-360 between healthy volunteers and patients with functional dyspepsia. The pharmacokinetic parameters showed that functional dyspepsia reduced the absorption of puerarin after oral administration of Jiawei-Xiaoyao-San. PMID- 19126308 TI - UK provision for removal of non-palpable contraceptive implants. PMID- 19126309 TI - Male circumcision, human papillomavirus and cervical cancer: from evidence to intervention. PMID- 19126310 TI - Use of contraceptive services in Britain: findings from the second National Survey of Sexual Attitudes and Lifestyles (Natsal-2). AB - OBJECTIVE: To describe contraceptive service use and identify demographic and sexual behavioural characteristics associated with use (and non-use) of different services. METHODS: Probability survey sample of men and women aged 16-44 years, resident in Britain. Participants comprised 3369 men and 4375 women reporting vaginal intercourse in the last year (excluding those reporting exclusive use of sterilisation or medical investigations for infertility). Main outcome measures were use of contraceptive services, grouped as: general practice, community contraceptive clinics, retail services and nonuse of services. RESULTS: General practice was the most commonly reported source of contraceptive supplies for women (59.2%), while retail services were most frequently reported by men (42.7%). 16.3% of women and 7.3% of men reported using more than one type of service. 20.7% of women and 45.1% of men had used no service in the last year, and amongst 16-17-year-olds the proportions reporting non-use of services was 13.8% and 31.2%, respectively. Use of community contraceptive clinics was associated with being younger, childless, single and reporting more heterosexual partners in the last year. CONCLUSIONS: There was relatively little 'shopping around' between different services, suggesting that choice of contraceptive providers ensures a range of needs is met for most people. While general practice is the most commonly used source of supplies, community contraceptive clinics are seeing those potentially at higher sexual health risk, particularly the young and those with multiple partners. Ways of improving young people's access to services for contraceptive supplies need to be addressed. PMID- 19126311 TI - Service provision for women undergoing termination of pregnancy: progress in Wales, UK. AB - BACKGROUND: This audit established a recent picture of termination of pregnancy services in Wales, UK using the Royal College of Obstetricians and Gynaecologists' Guidelines as a baseline. The context of abortion is rapidly changing and services need to be adaptable to meet women's needs. METHODS: A questionnaire survey was sent to all National Health Service (NHS) Trusts in Wales; 10 out of 13 responded. RESULTS: In the nine Trusts performing abortions in Wales, medical abortions accounted for 57% and surgical abortions for 43%. Doctors in training were involved in six Trusts. All but one Trust complied with referral times. Five Trusts provided a dedicated clinic. Written information provided prior to abortion varied in accessibility and quality. Choice of abortion within gestation bands was limited in some Trusts with some only providing medical termination. Essential abortion aftercare was performed by Trusts, whereas follow-up and counselling were less comprehensive. DISCUSSION: Trusts are willing to adapt to new methods of working with an increasing number of medical terminations, although this advantage was offset by a lack of choice of abortion methods offered by some Trusts. Doctors in training should be offered exposure to abortion procedures in all Trusts as this has been shown elsewhere to improve attitudes. Providing dedicated abortion clinics, quality written information and comprehensive abortion aftercare should improve compliance with the guidelines. CONCLUSION: Overall, in Wales there is compliance with national guidelines, although for a number of the guideline recommendations there remains room for improvement. PMID- 19126313 TI - Testing and quantifying association in binary data. PMID- 19126312 TI - Willingness of young men and women to be tested for Chlamydia trachomatis in three non-medical settings in Glasgow, UK. AB - OBJECTIVE: To assess the willingness of young men and women to be tested for Chlamydia trachomatis in three non-medical settings. METHODS: Men and women aged between 16 and 24 years were invited to complete a self-administered questionnaire and provide a urine sample in non-medical settings: 'education' (one further education college), 'health and fitness' (three local authority leisure centres) and 'workplace' (two call centres). RESULTS: Eighty-four percent of age-eligible users approached in the settings agreed to complete a questionnaire (n = 363). Among the sexually active people (n = 346), the uptake of screening varied by setting [education 19.1% (22/115), health and fitness 48.8% (62/127), workplace 27.8% (29/104); p<0.001]. Health and fitness settings (OR 4.08; 95% CI 2.04-8.14) and perception of being at risk of having chlamydia (OR 2.47; 95% CI 1.33-4.58) were strong predictors of providing a urine sample. Adjusting for setting and age group (<20 years vs 20+ years), women were less likely than men to provide a urine sample (OR 0.42; 95% CI 0.26-0.70). All five positive cases (4.4%; 4.9% in men, 3.8% in women) were contacted with their results by a health adviser and invited to be treated at a local genitourinary medicine clinic. CONCLUSIONS: Men were more willing than women to be tested for C. trachomatis in these non-medical settings, but uptake varied by setting. Thus, increasing opportunities for the take-up of testing in particular non-medical settings might be a more effective approach to including young men who are not reached by clinic control efforts. PMID- 19126314 TI - Human papillomavirus (HPV) information needs: a theoretical framework. AB - BACKGROUND: With the introduction of human papillomavirus (HPV) testing and vaccination in the UK, health professionals will start to receive questions about the virus from their patients. This study aimed to identify the key questions about HPV that British women will ask when considering having an HPV test or vaccination. METHODS: Face-to-face interviews were carried out with 21 women to discover what they wanted to know about HPV. A thematic framework approach was used to analyse the data and identify key themes in women's HPV knowledge requirements. RESULTS: Women's questions about HPV fell into six areas: identity (e.g. What are the symptoms?), cause (e.g. How do you get HPV?), timeline (e.g. How long does it last?), consequences (e.g. Does it always cause cervical cancer?) and control-cure (e.g. Can you prevent infection?). In addition, they asked procedural questions about testing and vaccination (e.g. Where do I get an HPV test?). These mapped well onto the dimensions identified in Leventhal's description of lay models of illness, called the 'Common Sense Model' (CSM). DISCUSSION AND CONCLUSIONS: These results indicated that the majority of the questions women asked about HPV fitted well into the CSM, which therefore provides a structure for women's information needs. The findings could help health professionals understand what questions they may be expected to answer. Framing educational materials using the CSM themes may also help health educators achieve a good fit with what the public want to know. PMID- 19126315 TI - Gynaecological ultrasound in community sexual and reproductive health clinics. AB - BACKGROUND AND METHODOLOGY: Many women seen in community sexual and reproductive health (SRH) clinics have gynaecological problems and a pelvic ultrasound scan forms part of their investigation. We present a retrospective analysis of 24 months' provision of a gynaecological ultrasound service in the Department of Sexual and Reproductive Health, Southwark Primary Care Trust, London, UK. RESULTS: A total of 327 women attended for ultrasound examination; 258 required a pelvic scan and 69 had an Implanon-related problem. Of the women analysed, 152 were managed entirely in the community; eight women required referral to gynaecological outpatients and five women needed an abdominal pelvic X-ray. CONCLUSIONS: We believe we have demonstrated that gynaecological ultrasound can be performed in the community. This seems to be acceptable to patients and also reduces the requirement for hospital appointments. PMID- 19126317 TI - Engaging with pre-teens and their parents in primary care. AB - OBJECTIVE: To encourage young people to think about health, including sexual health, and access primary care appropriately and confidently. CONCEPT: To engage with young people before they become teenagers. This is a single introductory session for 10-year-olds with their parents. The partly structured evening is held in the surgery after it closes for other business. It includes discussion about puberty, relationships and any other health matters raised by the participants. There is an introduction to the idea of patient confidentiality. RESULTS: About one-third of the invited 10-year-olds attended. Separating the genders improved attendance. Feedback suggested that young people increased their knowledge of puberty. They were more confident approaching health care staff for sexual health advice at the dedicated Young Person's Clinic or at normal surgery. Parents reported that their children talked more openly about relationships and sex following the session. CONCLUSION: A dedicated evening for pre-teens with their parents is worthwhile in primary care. PMID- 19126318 TI - Involving pharmacists in sexual health research: experience from an emergency contraception study. AB - BACKGROUND: Community pharmacists are expanding their sphere of activity within primary health care, increasing their role not only in health care but also research. METHODS AND RESULTS: We describe the challenges encountered in carrying out a pilot study of women obtaining emergency hormonal contraception through different providers, including pharmacies, highlighting deficiencies in understanding and experience of the research process, which impacted on the study in substantial ways. CONCLUSIONS: As pharmacists expand their role, training and professional development will need to be enhanced to support them in their contribution to health care and research. PMID- 19126319 TI - Hygroscopic dilator (Dilapan-S) and misoprostol combination for the early first trimester termination of pregnancy: a pilot study. AB - BACKGROUND AND METHODOLOGY: Uptake of the mifepristone/misoprostol combination to induce early medical abortion in England and Wales has been slow. Women's concern that early medical abortion is painful may be a contributory factor. This pilot study evaluated the pain experienced by women when Dilapan-S, a synthetic hygroscopic dilator (polyacrylonitrile) is used instead of mifepristone as cervical preparation prior to administration of a misoprostol. RESULTS: Of 25 patients completing the trial, 17 aborted in a median of 6 hours with an interquartile range of 4.5-11.5 hours. Of these, 15 patients recorded mild discomfort only, 14 considered the procedure excellent and three good. DISCUSSION AND CONCLUSIONS: This study suggests that the Dilapan-S/misoprostol combination reduces the pain associated with early medical abortion. Further investigation of the protocol is merited. PMID- 19126320 TI - Why lesbians should be encouraged to have regular cervical screening. AB - INTRODUCTION: The UK National Health Service Cervical Screening Programme (NHSCSP) advice on cervical cytology screening states that women who have never had sex with men are at very low risk of developing cervical cancer, and advice regarding need for screening in lesbians is inconsistent. METHODS: Literature review searching PubMed, Web of Science and the Internet for articles on lesbians, cervical cancer and cervical cancer risk factors focusing on human papillomavirus (HPV) and screening behaviours. RESULTS: Case reports and prevalence studies show that HPV can be transmitted sexually between women. It is not known whether prevalence of HPV or cervical cancer differs between lesbians and heterosexual women. The evidence consistently shows that prevalence of non attendance for cervical screening is much higher in lesbian than heterosexual women, which is linked to a belief that lesbians are less susceptible to cervical cancer and have less need for screening. Despite sharing most of the same risk factors as heterosexual women, lesbians are much less likely to undergo regular screening. CONCLUSIONS: The NHSCSP should take a clear and consistent stance on the need for cervical screening in lesbians. Both the health care and the lesbian communities must be made aware of the fact that regular cervical screening is as important in this group as it is in the heterosexual female population. PMID- 19126321 TI - Judaism and women's health. PMID- 19126322 TI - Mr Mom: can we deliver? PMID- 19126323 TI - Contraceptive failure of Depo-Provera: long-acting reversible contraceptive (LARC) methods do fail too. PMID- 19126324 TI - A partnership of equals. PMID- 19126325 TI - The Kinsey Institute. PMID- 19126326 TI - Positive smear and on: the partner's story. PMID- 19126327 TI - Sex and obscenity in medieval art. PMID- 19126329 TI - Anterior shoulder dislocation. AB - INTRODUCTION: Anterior dislocation of the shoulder is commonly seen in accident and emergency (A&E) and trauma clinics. In this article, we review the existing literature on the injury and the recent trends in management. MATERIALS AND METHODS: We have discussed this condition with our colleagues and performed a Medline search ('anterior shoulder dislocation') of the relevant papers. We also describe key historical publications and recent developments regarding immobilisation of the joint. RESULTS: Management decisions regarding this condition continue to vary between units, especially for recurrent and posterior dislocation. This paper lays some emphasis on the choice of analgesic agent when attempting shoulder reduction in the A&E setting. A summary of the data from our own department has provided a graphical representation of the classical age and sex distribution for this condition. CONCLUSIONS: Shoulder dislocation is a common injury. Delays in diagnosis remain the single biggest obstacle to optimum results in this group of patients. A significant proportion will require eventual surgery and up to a third of these patients will go on to develop long-term shoulder arthritis. Even patients who have experienced a single episode of dislocation may go on to develop long-term sequelae. PMID- 19126330 TI - Management of retrosternal goitres. PMID- 19126331 TI - The emergency surgical team - the way forward in emergency care? AB - INTRODUCTION: With reduced working hours and shift patterns, surgical training and continuity of patient care is being put at risk. We have devised a system for managing the emergency surgical patients in an effort to counteract these perceived problems. This study describes the emergency surgical team and audits its activity. PATIENTS AND METHODS: The emergency surgery team concept is described in detail. Over a 2-week period, general surgical referral data, patient management and operative activity were audited. RESULTS: A total of 229 patients were referred to the emergency surgical team with 159 treated conservatively, 45 underwent operative intervention and 25 were discharged without admission. Of the emergency surgical team referrals, 58% had gallstone pathology, appendicitis or constipation/non-specific abdominal pain. Average daily number of patients under the care of the emergency surgical team was 26 (range, 10-40). CONCLUSIONS: The consultant-led emergency surgical team look after many of the acutely sick surgical patients. Our system not only provides good teaching opportunities but ensures optimal continuity of patient care in a busy district general hospital. Such an approach to emergency surgical care has been successfully developed to optimise training opportunities and improve patient care in a setting of reduced working hours and shift systems in our hospital. PMID- 19126332 TI - A one-stop clinic for men with testicular anxiety. AB - INTRODUCTION: Testicular lumps and orchalgia both cause considerable anxiety, usually related to concerns about possible cancer. PATIENTS AND METHODS: We established a rapid-access testicular clinic staffed by a urologist and a specialist ultrasonographer in order to delay the time to a definitive diagnosis. RESULTS: Over a 30-month period, 845 men underwent clinical examination and scrotal ultrasonography. Overall, 4% of men were found to have testicular cancer. The majority of men were found to have normal testes or minor abnormalities and could be re-assured. CONCLUSIONS: This one-stop clinic reduces the time to a definitive diagnosis which may reduce anxiety suffered by men with testicular complaints and allows rapid identification of those harbouring a testicular malignancy. PMID- 19126333 TI - A simple prophylaxis regimen for MRSA: its impact on the incidence of infection in patients undergoing liver resection. AB - INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) infection has increased at an alarming rate in the recent past and has major cost implications. The aim of this study is to assess the impact of a policy of pre-operative MRSA prophylaxis on the incidence of MRSA infection in patients undergoing liver resection. PATIENTS AND METHODS: A total of 585 patients underwent hepatectomy in a tertiary referral centre between January 2000 and September 2005. In September 2003, a policy of MRSA prophylaxis (nasal mupirocin and triclosan wash for 5 days) was introduced within this unit. Demographic, pathological and outcome data were compared between the pre- and post-MRSA prophylaxis cohorts. RESULTS: The prevalence of MRSA infection prior to initiating the prophylaxis protocol was 29 patients (8.3%) and this fell to 9 patients (3.8%; P = 0.029). Furthermore, patients who had MRSA-related infection had a higher incidence of additional complications compared to the rest of the cohort (P = 0.001). Total cost savings incurred as a result of this protocol over the past 2 years has been approximated at 28,893 pounds. CONCLUSIONS: Introduction of a simple MRSA prophylaxis policy has had a significant reduction on the incidence MRSA-related infection within our patient population, leading to reduced morbidity and cost saving to the UK National Health Service. PMID- 19126334 TI - The cost of ignoring acute cholecystectomy. AB - INTRODUCTION: Biliary symptoms whilst awaiting elective cholecystectomy are common, resulting in hospital admission, further investigation and increased hospital costs. Immediate cholecystectomy during the first admission is safe and effective, even when performed laparoscopically, but acute laparoscopic cholecystectomy has only recently become increasingly commonplace in the UK. This study was designed to quantify this problem in our hospital and its cost implications. PATIENTS AND METHODS: The case notes of all patients undergoing laparoscopic cholecystectomy in our hospital between January 2004 and June 2005 were examined for details of hospital admissions with biliary symptoms or complications whilst waiting for elective cholecystectomy. Additional bed occupancy and radiological investigations were recorded and these costs to the trust calculated. We compared the potential tariff income to the hospital trust for the actual management of these patients and if a policy of acute laparoscopic cholecystectomy on first admission were in place. RESULTS: In the 18-month study period, 259 patients (202 females) underwent laparoscopic cholecystectomy. Of these, 147 presented as out-patients and only 11% required hospital admission because of biliary symptoms whilst waiting for elective surgery. There were 112 patients who initially presented acutely and were managed conservatively. Twenty four patients were re-admitted 37 times, which utilised 231 hospital bed-days and repeat investigations costing over 40,000 pounds. There would have been a marginal increase in tariff income if a policy of acute laparoscopic cholecystectomy had been in place. CONCLUSIONS: Adoption of a policy of acute laparoscopic cholecystectomy on the index admission would result in substantial cost savings to the trust, reduce elective cholecystectomy waiting times and increase tariff income. PMID- 19126335 TI - Stenting for large bowel obstruction - evolution of a service in a district general hospital. AB - INTRODUCTION: Stenting for obstructing large bowel malignancy is a technique that is gradually increasing in popularity. The two main indications are for palliation and as a 'bridge to surgery'. Some of the proposed advantages of colonic stenting are safety, reduced morbidity and mortality, avoidance of a stoma and shorter hospital stay. PATIENTS AND METHODS: This was a retrospective study of consecutive patients who had self-expanding metal stents deployed between February 2001 and June 2006. Data were collected from the MEDITECH electronic integrated healthcare information support system and case note review. Data concerning demographics, primary diagnosis, and location of malignant stricture, indication for stenting, method of stenting, outcome, complications and mortality rates were obtained and analysed on Microsoft Excel. RESULTS: Colonic stenting was first performed in the Countess of Chester Hospital in 2001. Thirty-two procedures have been performed since then. The median age was 80 years and the majority of cases were palliative (28 of 32), with three of the remaining cases successfully stented as a 'bridge to surgery'. Initially, this was performed as a radiological procedure; however, the success rate was noted to be better if a surgical endoscopist was also involved. We recorded a 57% clinical success rate in the group of patients that had the colonic stent inserted radiologically; however, the group where this was inserted as a combined radiological and endoscopic procedure yielded a clinical success rate of 78%. We experienced stent-migration in four patients (13%) and rectal perforation in one patient (3%). There was no tumour re-obstruction or stent-related mortality. CONCLUSIONS: A colonic stenting service can be introduced into a district general hospital with low morbidity and mortality. A well-motivated team is required and combined endoscopic and radiological approach in our hands appears to offer the best results. PMID- 19126337 TI - Community-based orthopaedic follow-up. Is it what doctors and patients want? AB - INTRODUCTION: The objective of this study was to investigate how patients, general practitioners (GPs) and orthopaedic trainees, feel about the proposed governmental changes to reduce orthopaedic out-patient clinics by having GPs and specialist nurses follow-up postoperative orthopaedic patients in the community. SUBJECTS AND METHODS: The design was a cross-sectional questionnaire study including a teaching hospital and general practitioners in the Norfolk primary care trust. Participants were 73 orthopaedic postoperative patients who attended out-patients over a 1-week period in July 2007 who all responded. Of 250 GPs, 239 responded. Of 38 orthopaedic trainees at the level of senior house officer (post MRCS) and specialist registrar (Eastern Deanery Rotation and Pott Rotation), 30 responded RESULTS: Of the 73 patients, 56 (77%) felt the surgeon was best suited to manage them postoperatively. Of these, 47 felt that it was very important that the surgical team saw them postoperatively. Also, 53 felt that their GP did not have sufficient knowledge and experience to deal with their current orthopaedic problem adequately. Only 12 GPs of 239 (5%) felt very confident assessing postoperative patients. Inadequate resources available to diagnose and treat postoperative complications was noted by 74% as the reason for not performing follow-up in primary care, and only 18% felt they should follow-up postoperative patients. All trainees felt that following up their own postoperative patients was important to their training. CONCLUSIONS: Most patients, GPs, and orthopaedic trainees had serious doubts about the proposed governmental changes to reduce orthopaedic out-patient clinics by having GPs and specialist nurses follow-up postoperative orthopaedic patients in the community. PMID- 19126338 TI - A strategy to overcome the radiology lottery in the staging of head and neck cancer: an aid to attaining the 30-day rule. AB - INTRODUCTION: Rapid access to radiological services is essential, if the British Association of Otolaryngologists - Head and Neck Surgeons Minimum Temporal Standards are to be met in the management of head and neck cancer patients. This study assesses a new initiative whereby the multidisciplinary team prioritises allocated imaging appointments rather than using the traditional radiological triage system. PATIENTS AND METHODS: This study was a prospective audit of all patients referred over a 3-month period with suspected head and neck cancer. The main outcome measures were: (i) median interval in days from general practitioner (GP) referral to staging scan; and (ii) median interval in days from first clinic appointment to staging scan. RESULTS: The new multidisciplinary team booking system led to a statistically significant reduction in the 'request-to-scan time' (from 12 days to 5 days). The time from 'GP to scan' also improved. CONCLUSIONS: This new multidisciplinary team-led booking system, could, in the future, speed up access to radiology services for head and neck cancer patients, allowing earlier definitive treatment. PMID- 19126339 TI - A helping hand for the lone surgeon fixing a lateral malleolus fracture. PMID- 19126340 TI - General paediatric surgery provision. PMID- 19126341 TI - Swallow the medicine. PMID- 19126342 TI - Innovation in surgery. PMID- 19126343 TI - Simulators in self-assessment. PMID- 19126344 TI - Plans for the future: a strategy for the Faculty of General Dental Practice (UK) in the next five years. PMID- 19126345 TI - Education for the dental team: make your practice a centre of learning excellence. AB - This paper discusses the future educational needs of the changing dental team, the potential for new institutions and teachers to deliver and assess that education, including outreach-based teaching and learning, and interchangeability of educational credits between institutions. The author's opinion is that primary care practices can become 'centres of learning' where all members of the dental team can interact to teach and learn together at the same time. The development of a career pathway for dental care professionals (DCPs), a more unified pan Europe approach to integrated dental education and the development of further roles for senior dental students are proposed. The importance of dentistry acting upon sound, innovative concepts, and of not resisting educational change, is stressed. PMID- 19126346 TI - Can protein contamination be removed from hand endodontic instruments? AB - AIM: The aim of this study was to quantify total protein adhering to hand endodontic files and to measure and compare the efficacy of ultrasonic cleaning and washer-disinfectors, with and without presoaking, in protein removal from clinically contaminated endodontic files. METHOD: Total protein contamination of the endodontic files was quantified using an assay reagent colorimetric method. Twelve general dental practitioners were recruited to collect clinically contaminated files. One hundred and fifty clinically contaminated files were allowed to air-dry in sterile plastic containers and a further 60 files were immersed, working end down, in enzymatic detergent immediately following clinical use. Thirty clinically contaminated files were tested for total protein contamination as a positive control. Sixty files were subjected to ultrasonic cleaning and 30 to processing in each of the washer-disinfectors. The presoaked files were divided into two groups of 30 for processing in the washer disinfectors. A further group of brand-new, unused files were tested for protein contamination as a negative control. RESULTS: Protein was present on 29 of the 30 new files tested. The median total mass of protein recovered from clinically contaminated hand endodontic instruments was 2.046 microg. The most effective method of presterilisation cleaning tested was a presoak in Alkazyme followed by processing in the Miele G7881 washer-disinfector. CONCLUSION: The most effective method of presterilisation cleaning for hand endodontic files is a presoak in Alkazyme, an alkaline enzymatic detergent, followed by processing in a Miele G7881 washer-disinfector. This study provides up-to-date evidence that newer methods of presterilisation cleaning may fail to remove protein from endodontic hand instruments totally. This may have implications for all reusable dental instruments. PMID- 19126347 TI - Factors affecting the referral of potentially malignant lesions from primary dental care: a pilot study in South Yorkshire. AB - AIMS: Early diagnosis of oral cancer is an essential element of prevention, yet many patients present late, suggesting that criteria used for referral miss the small or early-stage lesions. The aims of this study were to determine what cues are used by primary care dentists when making the decision to refer a suspicious lesion, and to investigate their recent referral behaviour. METHODS: A cross sectional study using a postal questionnaire was undertaken in a major conurbation in South Yorkshire. The questionnaire was sent to all 95 practices in the area concerned. Primary care dentists were asked to score a range of cues on a five-point scale and list their top ten. Experience of referrals made in the last 12 months was also recorded. RESULTS: Forty-two questionnaires were returned (44.2%). The cues that primary care dentists 'agreed' or 'strongly agreed' would initiate a referral were: 'fixed', 'smokes >20 cigarettes/day', 'indurated', 'ragged borders' and 'ulceration'. The most cited top-ten cues were 'ulcerated', 'fixed to underlying tissues', 'smoking' and 'indurated'. The median number of lesions referred was 0.77 per 1000 adult patients. Twenty-six dentists recorded their experience of 29 lesions. Of these, 17% had been found by the patient whereas 80% had been found at a routine dental examination. Seventy-nine per cent had been present for more than six weeks. Reasons for referral included atypical appearance (38%), ulceration (21%) and location (21%). Fifty-four per cent of primary care dentists described their relationship with secondary care as either 'good' or 'excellent'. CONCLUSIONS: The most cited cues related to established or advanced disease, not to 'red' and 'white' lesions, which the National Institute for Health and Clinical Excellence (NICE) recommend for early referral. Even allowing for the relatively poor response rate, this suggests that some small lesions of oral cancer are being missed. PMID- 19126349 TI - Developing consumer involvement in primary dental care. Report of a half-day seminar held at the Royal College of Surgeons of England on 15th September 2008. AB - The seminar on developing consumer involvement in primary dental care, held during the morning of 15th September 2008, was a collaboration between the Lay Advisory Group and Research Committee of the Faculty of General Dental Practice (UK) (FGDP[UK]). As Professor Mike Mulcahy (immediate past Dean of the Faculty) remarked during his address of welcome, it marked a new and exciting development in the Faculty's role in setting and maintaining professional standards for the benefit of patients. It brought together nearly 50 representatives of national bodies, such as the National Audit Office, consumer groups, the Faculty's Lay Advisory Group and Research Committee, the media and others. Many of the national bodies represented at the seminar had published reports on primary dental care during the last five years. PMID- 19126348 TI - Mentoring. A quality assurance tool for dentists. Part 2: what are mentoring and coaching? AB - In this paper, the traditional model of mentoring is briefly explained. A description of the current technique, which has developed from the traditional model, is then presented and a distinction made between mentoring and coaching, followed by a brief explanation of how mentoring, coaching and counselling make up a triad of helping activities. The authors then provide information on the use and impact of mentoring and coaching in some areas of human activity, other than dentistry, before outlining the application potential of these approaches in general dental practice. The paper concludes that the modern approach to mentoring and counselling offers a person-centred approach that is much more likely than traditional approaches to produce personal change, personal growth and personal development. Further aspects of this important and exciting subject will be explored in subsequent papers in this series. PMID- 19126350 TI - A question of value: a qualitative study of vocational dental practitioners' views on oral healthcare systems and their future careers. AB - BACKGROUND: New dental graduates in England and Wales spend one year as vocational dental practitioners (VDPs) preparing for independent clinical practice. In recent years, they have entered a state-funded healthcare system undergoing the greatest period of change since the inception of the National Health Service (NHS) and a profession in which there has been a significant shift of care to the private sector. Against this background, the objectives of this study were to investigate VDPs' vision of their future professional career and the influences that will impact on their choice of state-funded (NHS) and/or private dentistry, and to identify what factors may attract graduates to work for the NHS. METHODS: In 2004/2005, purposive sampling of a range of VDP training schemes across England and Wales was used to select the VDPs from ten schemes to take part in focus groups. To standardise data collection, a topic guide was used. Respondents' views were recorded on tape and field notes. The data were transcribed and analysed using framework methodology. RESULTS: Ninety-nine VDPs from all parts of England and Wales participated in ten focus groups. They identified three main categories of future practice: private, state-funded (NHS), and mixed. Private practice was perceived as providing 'professional independence', 'financial reward', 'time with patients' and 'clinical freedom'. NHS practice was associated with 'providing access to specialist training' and 'gaining clinical experience', often as preparation for private practice. Providing NHS care was attractive for VDPs who valued the ethos of public service. The VDPs considered that NHS practice could be made more attractive to young dentists by a range of factors, involving the funding, culture and philosophy of the system and the degree of fit with their personal and professional vision. They reported that they would welcome 'incentives to work in areas of high need', 'assistance with debt' and a 'culture of valuing NHS dentists'. CONCLUSIONS: The findings suggest that the commitment to healthcare systems of the VDPs who took part in this study was associated with being true to their values and being valued within the system. They perceived a tension between state-funded and private practice, considering the latter more likely to meet personal and professional expectations. However, they remained open to working in an enhanced and supportive state-funded system, should it correspond with their values, and demonstrate that they were valued healthcare professionals. PMID- 19126351 TI - University of Manitoba's Faculty of Dentistry--"traditions of excellence: horizons of change". PMID- 19126352 TI - Innovative preclinical dental teaching and learning at the University of Manitoba. PMID- 19126353 TI - Outreach and service learning: Manitoba's Centre for Community Oral Health. PMID- 19126354 TI - Undergraduate experience in dental research: the Bachelor of Science (Dentistry) program at the University of Manitoba. PMID- 19126355 TI - Electronic curriculum mapping: supporting competency-based dental education. PMID- 19126356 TI - What additional precautions should I take when bonding to severely fluorotic teeth? PMID- 19126357 TI - The state of oral health in personal care homes: a public health issue? PMID- 19126358 TI - Are Manitoba dentists aware of the recommendation for a first visit to the dentist by age 1 year? AB - BACKGROUND: The Canadian Dental Association (CDA) and the American Academy of Pediatric Dentistry (AAPD) recommend that children visit the dentist by 12 months of age. PURPOSE: To report on how Manitoba"s general dental practitioners and pediatric dentists manage oral health in early childhood. METHODS: Mailed surveys that used the modified survey methods of Dillman were sent to 390 Manitoban general dental practitioners and pediatric dentists. The sampling frame was the Manitoba Dental Association"s Membership Registry, but only those dentists who consented to the release of their mailing information were contacted. Survey data were analyzed with Number Cruncher Statistical Software (NCSS 2007). Descriptive statistics, bivariate analyses and multiple regression analyses were done. A p value of < or = 0.05 was considered statistically significant. RESULTS: A total of 292 (74.9%) of the 390 practitioners responded, of whom 85.1% met the eligibility criteria and 84.6% were graduates of the faculty of dentistry, University of Manitoba. Overall, infants and preschoolers constituted < 10% of patients in the respondents" practices. Slightly more than half (58.3%, 144/247) of participants were aware of professional organizations" recommendation about the timing of children"s first visit to the dentist; 52.2% (130/249) were unaware of the existence of a standardized case definition for ECC; and 32.3% (80/248) knew that ECC was defined as the presence of at least 1 primary tooth affected by caries in children < 6 years of age. On average, these participating dentists from Manitoba thought children should visit the dentist by 2 years of age. CONCLUSIONS: Although early visits to the dentist are now endorsed by CDA and AAPD, a significant number of dentists in Manitoba are still unaware of the recommendation that children should first visit the dentist by 12 months of age. PMID- 19126359 TI - Tobacco use cessation services provided by dentists and dental hygienists in Manitoba: part 1. Influence of practitioner demographics and psychosocial factors. AB - OBJECTIVE: Despite high rates of tobacco use, overwhelming evidence of detrimental effects on oral health, smokers" desire to stop using tobacco and the availability of efficacious brief intervention counselling (BIC) strategies, the delivery of cessation services by dental practitioners is, at best, inconsistent. The purpose of this part of our study was to assess BIC practice patterns among dentists and dental hygienists in Manitoba and to determine whether demographic or psychosocial factors influence BIC delivery. METHODS: A pre-piloted survey was mailed to all licensed dentists (547) and registered dental hygienists (566) in the province. RESULTS: In all, 514 oral health practitioners responded for a 46.2% response rate. Most oral health practitioners in Manitoba are not providing consistent BIC; however, 54.9% (279/508) of survey respondents advise smokers to quit. Women clinicians are more likely to ask, assess and assist patients and tend to advise against smoking more frequently than men; younger practitioners are more likely to ask and assess readiness to quit smoking than older practitioners; dental hygienists are more likely to provide assistance to quit than dentists. Assisting is the service least frequently provided by practitioners. The barriers to providing BIC are different for male and female practitioners and for dentists and dental hygienists; practitioners with more psychosocial barriers provide BIC less frequently than those reporting fewer barriers. Only 36.9% (188/510) of practitioners report feeling adequately prepared to assist smokers to quit. PMID- 19126360 TI - Three-way trays: easy to use and abuse. AB - The 3-way tray technique is popular as it provides master and opposing arch impressions and an interocclusal record at the same time. Excellent clinical results can be achieved with appropriate case selection, use of rigid tray and impression materials, attention to tray positioning and other details throughout the process, and clear understanding of the limitations of the technique by the operator and dental technician. PMID- 19126361 TI - Shedding new light on early caries detection. AB - Dental caries continues to be a common chronic disease among various population groups. Patient care can be improved with detection at the earliest stage. However, current techniques do not have sufficient sensitivity and specificity. We discuss 2 new methods - optical coherence tomography (OCT) and polarized Raman spectroscopy (PRS) - that are potentially useful for early caries detection and monitoring. OCT produces morphologic depth images of near-surface tissue structures with a resolution that is an order of magnitude greater than ultrasound imaging. Based on measurement of back-scattered near infrared light, OCT shows that sound enamel causes high-intensity back-scattering at the tooth surface that decreases rapidly with depth. In contrast, incipient lesions cause higher light back-scattering at the tooth surface and subsurface scattering indicative of porosity caused by demineralization. The scatter region within the enamel correlates well with the classical triangular shape of subsurface lesions observed in histologic sections. OCT imaging not only allows identification of incipient lesions, but also provides information on surface integrity and lesion depth. PRS furnishes biochemical information about the tooth"s composition, mineral content and crystallinity. The depolarization ratio derived from the dominant phosphate peak of hydroxyapatite in sound teeth is consistently lower than that from incipient caries. This difference is attributed to the change in enamel crystallite morphology or orientation that occurs with acid demineralization. Thus, PRS can be used to confirm suspect lesions determined by OCT and rule out false-positive signals from non-carious anomalies. The combination of OCT and PRS provides a new detection method with high sensitivity and specificity that will improve caries management and patient care. Future studies are aimed at developing intraoral probes to validate the findings in vivo. PMID- 19126362 TI - Spinal cord stimulation compared with medical management for failed back surgery syndrome. PMID- 19126363 TI - Treatment of hemicrania continua by occipital nerve stimulation with a bion device. PMID- 19126364 TI - Factors contributing to pain chronicity. AB - The chronicity of pain is the feature of pain that is least understood and most directly linked with our inability to effectively manage pain. Acute pain is relatively responsive to our current pharmacologic and interventional armamentarium. However, as pain persists, our ability to treat effectively diminishes and the patient's frustration and resource utilization increases. This article explores our current understanding of the factors linked to pain duration and the transition from acute to chronic pain in both human and animal models, and across a spectrum of human chronic pain conditions. PMID- 19126365 TI - Noninvasive transcranial brain stimulation and pain. AB - Transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS) are two noninvasive brain stimulation techniques that can modulate activity in specific regions of the cortex. At this point, their use in brain stimulation is primarily investigational; however, there is clear evidence that these tools can reduce pain and modify neurophysiologic correlates of the pain experience. TMS has also been used to predict response to surgically implanted stimulation for the treatment of chronic pain. Furthermore, TMS and tDCS can be applied with other techniques, such as event-related potentials and pharmacologic manipulation, to illuminate the underlying physiologic mechanisms of normal and pathological pain. This review presents a description and overview of the uses of two major brain stimulation techniques and a listing of useful references for further study. PMID- 19126366 TI - Spinal cord stimulation for the treatment of angina and peripheral vascular disease. AB - Spinal cord stimulation has been shown to be an effective and well-established treatment for failed back surgery syndrome, complex regional pain syndrome, and other neuropathic pain states. Recent advances in this therapy have led to its use in enhancing blood flow and reducing ischemic pain patterns. The application of spinal cord stimulation to treat angina and improve outcomes in patients suffering from peripheral vascular disease is now becoming a part of the algorithmic standard of care. This article examines the selection of patients, application of the therapy, outcomes, and future uses of stimulation for patients afflicted with these diseases. This article also examines possible study protocols to further examine the overall outcome of these therapies. PMID- 19126367 TI - Methadone: a new old drug with promises and pitfalls. AB - Although methadone is not a new medication, its use in pain management has increased rapidly over the past decade. This article reviews the unique pharmacologic properties of methadone, including its long-acting nature, highly variable clearance rate, and its antagonism of the N-methyl-D-aspartate receptor. We discuss potential benefits and risks of methadone over other opioid medications. Preclinical studies suggest methadone may reduce abuse potential, tolerance development, and sensitization of nociceptive pathways. Pharmacologic properties of methadone suggest potential greater risk of dangerous or fatal side effects from overdose, QT interval prolongation, and drug interactions. However, clinical studies have yet to confirm that methadone produces either better clinical outcomes or higher rates of adverse events than other opioid analgesics. Clinicians who understand the special properties of methadone and follow recommended precautionary prescribing and monitoring practices can safely and effectively use methadone for pain treatment. PMID- 19126368 TI - Epidural steroid injections are useful for the treatment of low back pain and radicular symptoms: pro. AB - Epidural steroid injection has been used to treat low back pain for many decades. Numerous randomized trials have examined the efficacy of this approach. This review details the findings of older systematic reviews, newer randomized controlled trials, and two recent systematic reviews that examine the effectiveness of this treatment. Collectively, studies in acute radicular pain due to herniated nucleus pulposus have failed to show that epidural steroid injection reduces long-term pain or obviates the need for surgery. Similarly, there is scant evidence that epidural steroids have any beneficial effect in those with acute low back pain without leg pain or in those with chronic low back or leg pain. However, most studies have demonstrated more rapid resolution of leg pain in those who received epidural steroid injections versus those who did not. The role of epidural steroid injections in the management of acute radicular pain due to herniated nucleus pulposus is simply to provide earlier pain relief. PMID- 19126369 TI - Epidural steroid injections are useful for the treatment of low back pain and radicular symptoms: con. AB - Lumbar epidural steroid injections are commonly performed in the United States for treating radicular low back pain. However, the best available data suggest that the benefit afforded by these injections is quite limited; in fact, new data suggest that in geographic areas where many such injections are performed, more and not fewer spine surgeries are actually completed annually. We suggest that further high-quality studies are required and their results respected through their implementation in daily practice to better ensure that only appropriate patients are advised to undergo this procedure. PMID- 19126370 TI - Pain outcomes: a brief review of instruments and techniques. AB - Pain is a difficult outcome to measure due to its multifaceted and subjective nature. The need for selecting proper outcome measures is high because of the increasing demand for scientifically valid demonstrations of treatment efficacy. This article discusses some basic topics in the measurement of pain outcomes and addresses issues such as statistical versus clinical significance, daily home data collection, appropriate length of outcome measurement packets, and the possibility of objective pain measurements. This article also reviews some of the more commonly used tools for measuring pain and pain-related disability. By selecting the proper tools and employing them correctly, we can obtain highly reliable and valid measures of pain outcomes in research and clinical care. PMID- 19126371 TI - New daily persistent headache in children and adults. AB - New daily persistent headache (NDPH) is frequently seen in young patients with chronic daily headache. NDPH begins with a sudden onset, often associated with an infection or other physical stress. This headache syndrome is difficult to treat and may persist for years. This review discusses the epidemiology, comorbid symptoms, evaluation, and treatment of this disorder. PMID- 19126372 TI - Headache subsequent to whiplash. AB - Methodology varies greatly in whiplash studies; therefore, results are not directly comparable. Headache seems to be present in 50% to more than 75% of cases in the acute stage, and in 20% to 30% of cases in the early chronic stage. Headache naturally occurs frequently (> 75% of the cases ) in patients who consult headache specialists due to protracted symptoms. Malingering may explain some cases, and continuation/activation of prewhiplash headache may explain other cases. De novo headache also seems to occur. This headache may partly have cervicogenic headache characteristics: side-locked unilaterality and occipital onset. In one study, this type of headache was present in 8% at 6 weeks and 1% at 6 years. It was more rare than postwhiplash, unspecified headache. De novo postwhiplash headache may consist of cervicogenic headache and of noncervicogenic headache (probably in the acute phase). PMID- 19126373 TI - The epidemiology of chronic daily headache. AB - Chronic daily headache (CDH) is a fairly common but disabling disorder that disproportionately affects women and afflicts individuals across all stages of adulthood. It is a dynamic disorder, marked by relatively high rates of remission and incidence. To some extent, this may be due to the accepted, but not empirically supported, cut-point of 15 headache days per month. The purpose of this article is to understand the CDH classification; determine the prevalence and associated demographic profile of CDH as derived from population-based studies; outline identified risk factors for development or persistence of CDH; and understand which risk factors may be more amenable to intervention. Understanding the factors that put people at risk for developing CDH helps to inform possible clinical interventions and also determines which individuals may be most in need of preventive efforts. PMID- 19126374 TI - Chronic migraine: current pathophysiologic concepts as targets for treatment. AB - Chronic daily headache (CDH) affects approximately 4% of the population and exerts a significant degree of disability on its sufferers. Chronic migraine (CM) is a subset of CDH that represents migraine without aura occurring on 15 or more days per month for at least 3 months. Although numerous risk factors are associated with the development of CM, the pathophysiology governing its genesis is largely unknown. The role of neurotransmitters, such as glutamate, as well as disruptions of antinociceptive systems and structures, are implicated in CM and are supported by the fact that treatments targeting these abnormalities are effective. PMID- 19126375 TI - Chronic pain rehabilitation in chronic headache disorders. AB - This article reviews the treatment goals and efficacy of comprehensive pain rehabilitation programs for the treatment of chronic headache. Substantial data demonstrate improved outcomes from rehabilitative treatment for chronic noncancer pain. We present a discussion of the most relevant recent publications on pain rehabilitation in chronic headache disorders. This article describes pain rehabilitation, reviews outcome data for chronic pain patients treated in this setting, and describes the unique applicability of this treatment approach for patients with chronic headache. Particular attention is directed to the rationale for and the results of the withdrawal, in a pain rehabilitation setting, of opioids and simple analgesics, ergots, and triptans that contribute to medication overuse headaches. Additionally, a case example is reviewed that illustrates the structure and function of a pain rehabilitation program in the treatment of a patient with intractable headache. PMID- 19126376 TI - Clinical aspects of perimenstrual headaches. AB - Menstrual migraine (MM) is either pure, if attacks are limited solely during the perimenstrual window (PMW), or menstrually related (MRM), if two of three PMWs are associated with attacks with additional migraine events outside the PMW. Acute migraine specific therapy is equally effective in MM and non-MM. Although the International Classification of Headache Disorders-II classifies MM without aura, data suggest this needs revision. The studies on extended-cycle oral contraceptives suggest benefits for headache-prone individuals. Triptan mini prophylaxis outcomes are positive, but a conclusion of "minimal net benefit compared to placebo" is not entirely unwarranted. In a 2008 evidence-based review, grade B recommendations exist for sumatriptan (50 and 100 mg), mefenamic acid (500 mg), and riza-triptan (10 mg) for the acute treatment of MRM. For the preventive mini-prophylactic treatment of MRM, grade B recommendations are provided for transcutaneous estrogen (1.5 mg), frovatriptan (2.5 mg twice daily), and naratriptan (1 mg twice daily). PMID- 19126377 TI - Perimenstrual headache: treatment options. AB - At least half of women migraineurs experience menstrual migraine (MM), suggesting a hormonal explanation for the incidence of these headaches. Basic science efforts suggest a relationship between estrogen and the neurotransmitters and neuronal structures critical in the pathophysiology of migraine. The notion that MM is more severe, longer in duration, and more resistant to treatment than headaches occurring at other times during the menstrual cycle may apply more to women seeking treatment for their headaches than to migraineurs in the general population. Triptans have been shown to be effective as both an abortive and short-term preventive treatment, and estradiol has been shown to be an effective short-term preventive treatment. Ergotamines, combinations of drugs such as sumatriptan-naproxen sodium, and rizatriptan with dexamethasone show promise in the treatment of MM. PMID- 19126378 TI - [Construction and identification of cDNA library for colorectal carcinoma antigens]. AB - AIM: To construct a cDNA phage expression library of colorectal carcinoma antigens for screening colorectal carcinoma specific-antigens. METHODS: The total RNA was extracted from two colorectal carcinoma cell lines CCL-187 and CX-1 . The mRNA from total RNA was isolated to synthesize the first and second strand cDNA. The ds-cDNA termini were blunted with pfu DNA polymerase. The blunted cDNAs were ligated with EcoR I adapters and then digested by Xho I. Small cDNA molecules (<400 bp) were removed through size fraction. After the cDNAs were inserted into ZAP expression vector, the ligated products were packaged in vitro and the bacteriophage particles infected the host strains XL1-Blue MRF'. RESULTS: The efficiency of the primary was 2.3x10(9) pfu/L, while the recombination rate reached to 97%. The average length of the inserted fragment was over 1 kb. CONCLUSION: The quality of colorectal carcinoma cDNA phage expression library is excellent and helpful to screen colorectal carcinoma specific-antigens. PMID- 19126380 TI - [Establishment of a B16 cell line stably expressing genes encoding HER2 multi epitope peptides in vivo]. AB - AIM: To establish a B16 cell line stably expressing genes encoding HER2 multi epitope peptides in vivo. METHODS: Eukaryotic expressing vector pcDNA3-GFP-HER2 was constructed by molecular cloning technique and transfected into B16 cells mediated by cationic liposome. After screened with G418, the transfected cells were passaged in vivo. The GFP-HER2 positive cells were isolated from tumor burdening mice by flow cytometry (FCM) sorting, and then monoclonized in the absence of G418 to obtain a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo. This cell line was then identified after being passaged in vivo. RESULTS: Restriction endonulease analysis and DNA sequencing showed that the pcDNA3-GFP-HER2 was constructed and a B16 cell line stably expressing genes encoding HER2 multi-epitope peptides in vivo was obtained successfully. The GFP-HER2 positive proportion maintained higher than 90% after being passaged in vivo. CONCLUSION: A B16 cell line stably expressing genes encoding HER2 multi-epitope peptides is established successfully, which would provide a method to establish other cell lines stably expressing exogenous genes. PMID- 19126379 TI - [Expression of cytokines in murine BMDCs induced by recombinant E.coli LLO/OVA via TLR4 and NOD1 receptors]. AB - AIM: To investigate the relationship between the activation of pattern recognition receptors and the cytokines expression of dendritic cells. METHODS: After bone marrow-derived dendritic cells (BMDCs) were pulsed by E.coli LLO/OVA, the mRNA expression of pattern recognition receptors and the downstream NF-kappaB signal pathway associated molecules were detected by microarray hybridization and RT-PCR; the expression of costimulatory molecules, MHC class II and cytokines were determined by flow cytometry and ELISA. RESULTS: After BMDCs were pulsed by E.coli LLO/OVA for 2 h, the levels of TLR4, Myd88, Rip2, Irak1, Irak2, Ikkalpha, NF-kappaB1 and NF-kappaB2 mRNA up-regulated; and the levels of Icam1, IL-1a, IL 1b, IL-6 and TNF-alpha mRNA up-regulated also; after BMDCs were pulsed by E.coli LLO/OVA for 4 h, the levels of Card4(Nod1), Rip2, Ikkbeta, NF-kappaB1 and NF kappaB2 mRNA up-regulated, meanwhile, the levels of TNF-gamma, TNF-beta and CD40 mRNA up-regulated. The expressions of costimulatory molecules and MHC class II of the BMDCs up-regulated and the concentration of IL-12 and IFN-gamma increased in the supernatant of BMDCs pulsed by E.coli LLO/OVA at 24 h. CONCLUSION: Recombinant E.coli LLO/OVA induces maturation and expression of cytokines, especially IL-12 and IFN-gamma, of murine BMDCs through activation of NF-kappaB signal pathway with TLR4 and NOD1 receptors. PMID- 19126381 TI - [Tyrosine kinases inhibitor A77 1726 inhibits upregulation of collagen generation in fibroblasts induced by IL-13]. AB - AIM: To observe the inhibitory effects of tyrosine kinases inhibitor A77 1726 on collagen generation induced by IL-13 in fibroblasts. METHODS: The inhibition rate of fibroblast proliferation with different concentration of A77 1726 was observed by MTT method. The fibroblasts were divided into the experimental group (A77 1726 50 micromol/L and IL-13 100 microg/L) and the control group (IL-13 100 microg/L). After 24, 48 and 72 hours, the inhibitory effects of A77 1726 on collagen secretion of fibroblasts investigated by hydroxyproline release assay. The inhibitory effects of A77 1726 on collagen type I alpha1 gene mRNA expression in fibroblasts were examinated by RT-PCR. The influence of A77 1726 on collagen type I synthesis in fibroblasts was analyzed by Western blot. RESULTS: The proliferation of fibroblasts was inhibited by A77 1726. Total collagen generation was down-regulated after 48 h and 72 h stimulation of A77 1726 (P<0.05). The expression level of collagen type I alpha1 gene mRNA was obviously lower in the experimental group than in the control group after 48 h and 72 h stimulation of A77 1726 (P<0.05). Collagen type I production of fibroblasts treated with A77 1726 for 48 and 72 hours was decreased obviously in the experimental group than in the control group (P<0.05). CONCLUSION: Tyrosine kinases inhibitor A77 1726 has an inhibitory effect on collagen protein synthesis in fibroblasts induced by IL-13. PMID- 19126382 TI - [Application of Epstein-Barr virus-transformed B lymphoblastic cells in identification of CTL epitopes specific for Hantaan virus]. AB - AIM: To establish Epstein-Barr virus (EBV)-transformed B lymphoblastic cell lines (B-LCL) to present peptides as antigen-presenting cells (APC) and stimulate short cultured T cells secreting IFN-gamma, by which the T cell epitopes are identified. METHODS: PBMCs from patients with hemorrhagic fever with renal syndrome (HFRS) were transformed using EBV from supernatant of B95-8 cells. ELISPOT assay was then employed to evaluate the IFN-gamma production of short cultured G9L-specific CD8(+) T cells stimulated with peptide-pulsed autologous B LCL cells. RESULTS: B-LCL pulsed with G9L or G9L-nested V15R can stimulate G9L specific CD8(+) T cells producing IFN-gamma, but not B-LCL pulsed with non homologous I15P. CONCLUSION: B-LCL can efficiently and specifically present peptides to peptide-specific T cells as non-professional APC. PMID- 19126383 TI - [Effect of Rapamycin and Cyclosporin A on the expression of TLR5 and Foxp3 in allorejection]. AB - AIM: To study the effect of Rapamycin (RAPA) and Cyclosporin A (CsA) on the expression of TLR5 and Foxp3 in allotransplantation model in vivo. METHODS: The murine model of skin allotransplantation was established, and divided into three groups, injected with CsA 10 mg/(kg.d), RAPA 1.5 mg/(kg.d)) and normal saline respectively for 14 consecutive days. The grafts survival was observed daily. The expression of TLR5 and Foxp3 mRNA was detected by RT-PCR. RESULTS: The survival time was obviously longer in RAPA and CsA treated groups than control group. The expression of TLR5 mRNA in allotransplantation treated with RAPA was higher than that in control group(P<0.05) and remained at a higher level than the other two groups until day 21. TLR5 showed the highest expression on day 10 among three groups, which was the top point of allorejection. Compared with CsA group, RAPA treatment in vivo caused the higher expression of Foxp3 mRNA(P<0.05), which remained at a high level after treatment stopped. However, in CsA group, Foxp3 mRNA expression increased on day 7 and 10, and then decreased significantly on day 14. TLR5 and Foxp3 expression was positively correlated among three groups. RAPA and CsA promoted the expression of TLR5 in T cells when treated with flagellin for 6 hours in vitro and RAPA has stronger effect. CONCLUSION: RAPA and CsA can promote the expression of TLR5 and Foxp3 in allotransplantation model in vivo and flagellin enhanced this effect in vitro. PMID- 19126384 TI - [Jak/STAT signaling pathway of IL-11 in the protection of intestinal epithelial cells from neutron radiation]. AB - AIM: To explore the effect of IL-11 on the activation of Jak/STAT pathway and the expressions of Bax and Bcl-2 in the intestinal epithelial cells exposed to neutron radiation. METHODS: The BALB/c mice and IEC-6, irradiated by 4 Gy neutron with or without IL-11 treatment, served as in vivo and in vitro model seperately. The changes of the intestines, activity of Jak1 and STAT3 and expressions of Bax and Bcl-2 were observed by HE staining, Western blot, EMSA, immunohistochemistry and image analysis. RESULTS: (1)Mice exposed to neutron radiation showed severe intestinal damages and no obvious regeneration was seen. IL-11-treated mice had a larger number of cryptal epithelial cells and crypts. (2)Neutron radiation decreased the activities of Jak1 and STAT3, while IL-11 increased their activities. (3) Neutron radiation decreased the expression of Bax and didn't change the level of Bcl-2 in the murine intestine. IL-11 administration decreased the expression of Bax and increased that of Bcl-2. CONCLUSION: The mechanism of the intestinal protection of IL-11 in neutron irradiation might be that IL-11 stimulation triggered activation of Jak/STAT pathway, downregulated the expression of Bax and upregulated the expression of Bcl-2. PMID- 19126385 TI - [Effect of apoptotic ovarian cancer cells induced by paclitaxel-cisplatin on antigen presentation function of DC]. AB - AIM: To explore wheather apoptotic ovarian cancer cells induced by paclitaxel cisplatin could be cross-presented by antigen presenting cells and promote immune responses. METHODS: DCs were generated from peripheral blood monocytes in RPMI1640 supplemented with GM-CSF and IL-4. After 6 days' incubation, DCs were further co-cultured with either apoptotic HO8910 cell lines induced by paclitaxel cisplatin or control cells for four hours. Maturation of DCs was compared among different groups according to their surface markers through flow cytometry assay and their phagocytosis ability was evaluated by confocal microscopy scanning assay. RESULTS: Apoptotic ovarian cancer cells induced by paclitaxel-cisplatincan were phagocytized more efficiently by DCs.Resulting is more cellularity and maturation of DCs in apoptotic cell-induced groups than control group (P<0.05). CONCLUSION: Apoptotic ovarian cancer cells induced by paclitaxel-cisplatin exhibit strong immunogenicity, which in turn promote the maturation and antigen presentation of DCs. PMID- 19126386 TI - [B cell activation in experimental autoimmune myasthenia graves treated with Talpha146-162-iMDCs]. AB - AIM: To explore the therapic effect of Talpha146-162- iMDCs in C57BL/6 mice with experimental autoimmune myasthenia graves (EAMG), and illustrate whether this therapic effect is related with the change of B cells activation. METHODS: Adult male C57BL/6 mice were randomly divided into EAMG groups(A), the prevention group(B) and control group(C); Immature bone marrow dendritic cells were cultured and pulsed with Talpha146-162. Mice of A group and B group were evaluated for clinical score till the day they were put to death. The expression and phosphorylation of Syk, Lyn, Btk and PLC-gamma2 protein were measured by Western blot. RESULTS: 75% mice of A group and 25% mice of B group were developed the accumulated incidence, the difference was significant (P<0.05). The average clinical score of A group and B group were 1.69+/-1.12 vs 0.35+/-0.67(P<0.01) at the termination of experiment. The expression and phosphorylation of Syk and PLC gamma2 protein in spleen and lymphonode of the mice of A group were higher than those of C group (P<0.01) and B group (P< 0.05). Compared with C group, those of B group were higher (P<0.05); The expression and phosphorylation of Lyn protein in spleen and lymphonode of the mice of A (P<0.01) and B (P<0.05) groups were lower than those of C group. Compared with A group, those of B group were higher (P<0.05); The expression of Btk protein in spleen and lymphonode of the mice of A (P<0.01) and B (P<0.05) groups were higher than those of C group. And those of B groups were lower than those of A group (P<0.05). But there were no remarkable differences among the phosphorylation of Btk protein of three groups. CONCLUSION: Talpha146-162-iMDCs can prevent EAMG and probably ameliorate EAMG by the negative regulation on BCR signaling. PMID- 19126387 TI - [Effects of TWEAK on the synthesis of MMP-3 in fibroblast-like synoviocytes of rheumatoid arthritis]. AB - AIM: To investigate the effects of TWEAK on the synthesis of MMP-3 in RA FLS at different concentrations and to discuss the relative mechanism of how TWEAK involves in the destruction of articular bone and cartilage. METHODS: RA FLS were primarily cultured and stimulated with TWEAK. ELISA was used to detect the concentration of MMP-3 in cell-cultured fluid. The gene mRNA expression of MMP-3 was measured by RT-PCR. RESULTS: The level of MMP-3 induced by TWEAK at 50 and 100 microg/L was higher than that in control group, which had significant statistic difference (P<0.05).The expression level of MMP-3mRNA induced by TWEAK at 100 microg/L was 1.26 times higher than that in the control group, which had significant statistic difference (P<0.05).TNF-alpha and IL-1beta had synergetic effect on the synthesis and mRNA expression of MMP-3. The level in the synergetic group was significantly higher than that in the simple TWEAK group , which had significant statistic difference (P<0.05). CONCLUSION: TWEAK can induce RA FLS to synthesize MMP-3 and damage the articular bone and cartilage directly. TNF-alpha, IL-1beta and TWEAK had synergetic effects during the synthesis of MMP-3 in RA FLS. PMID- 19126388 TI - [Experimental studies on anti-mouse hepatocellular carcinoma effects of cisplatin combined with exosomes]. AB - AIM: To study the anti-tumor effects and mechanisms of cisplatin(DDP) combined with exosomes. METHODS: The cell-growth inhibition of DDP on H(22); cells was analyzed by MTT assay.The mice were immunized by H(22); cell-derived exosomes. The exosomes elicited CTL-specific cytotoxicity and DDP enhanced cytotoxicity in combination with CTL were detected by (3)H-TdR release assay. The effect of DDP on mRNA and protein levels of Fas in H(22); was analyzed using RT-PCR and Western blot.The expression of FasL on spleen lymphocyte was determined by RT-PCR. BALB/c mice inoculated with hepatoma carcinoma cell line H(22); were used as tumor models.The mice received exosomes solely or in combination with DDP.The survival of the mice was observed. RESULTS: DDP inhibited H(22); cell in a dose-dependant manner. The exosomes elicited CTL-specific cytotoxicity was enhanced by DDP (P<0.05). RT-PCR and Western blot showed Fas increased gradually after administering DDP on H(22); cells.RT-PCR also indicated the mRNA level of FasL on mice spleen lymphocyte increased after immunized by exosomes. Compared with other groups, the combination group(DDP plus exosomes)could statistically prolong the survival time (P<0.05). CONCLUSION: The therapy of DDP combined with exosomes had significant synergistic effect against tumor. The mechanism of synergistic effect includes enhancement of CTL activity. PMID- 19126390 TI - [Generation of human Fab antibody against platelet membrane glycoprotein IIb/IIIa and its effect on platelet aggregation]. AB - AIM: To generation of human Fab fragment against GP GPIIb/IIIa from phage display library and to study its effect on platelet aggregation. METHODS: An antibody phage display library was constructed from spleen cells from a donor with idiopathic thrombocytopenic purpura(ITP) whose anti-GPIIb/IIIa antibody was positive. High-affinity human mAbs was selected by panning against GPIIb/IIIa expressed on CHO123 cells. The binding specificity of phage antibody to GPIIb/IIIa was detected by ELISA and Western blot. And the effect of antibody on platelet aggregation was analyzed. RESULTS: After three rounds of panning with CHO123, phage antibodies against GPIIb/IIIa were enriched specifically. 2 positive phage antibodies with high specific for GP IIb/IIIa were verified, and the purified antibody can inhibited ADP induced platelet aggregation. CONCLUSION: Human antibodies against GPIIb/IIIa are obtained from phage display library. PMID- 19126389 TI - [Prokaryotic expression, polyclonal antibody preparation and subcellular localization of CIB]. AB - AIM: To construct the prokaryotic expression vector pET32a-CIB, prepare the specific polyclonal antibody against CIB and study the subcellular localization of CIB. METHODS: CIB was amplified by RT-PCR from human brain tissue and cloned into prokaryotic expression vector pET32a-CIB. The CIB fusion protein was expressed in BL21 (DE3)/pET system and identified by SDS-PAGE. The mice were immunized with the polyacrylamide gel particles containing the CIB fusion protein for polyclonal antibody preparation. The antibody was purified by affinity chromatographic column matrix coupled with protein G, antigen respectively and then identified by Western blot and immunohistochemistry. RESULTS: The protein of CIB was obtained by recombination expression. The specificity of polyclonal antibody was obtained by immunizing BALB/c mice with polyacrylamide gel particles containing the fusion protein of CIB and purification. The results of immunohistochemistry demonstrated that CIB was localized predominantly in the cytoplasm of SHG44 and Hhu7 cells. CONCLUSION: The protein of CIB has been cloned and expressed successfully. The specific polyclonal antibody against the protein of CIB has been obtained, which can be used for further research into the function of CIB. PMID- 19126391 TI - Regulation for chronic disease control: the pathfinder role of tobacco. AB - Regulation for health in the modern era has its foundations in the English Public Health Act of 1848. Early legislation was concerned with controlling environmental causes of disease. However, the focus on regulation today within health departments has diminished, displaced by a focus on services and related programs. The regulatory debate is now centred on what degree of protection, or safety margin, is required, and how regulatory efficiency may be improved. The example of tobacco control is reviewed to show how regulation can play a large role in chronic disease control, and consideration given to how regulatory tools could be further diversified and regulatory effectiveness improved. PMID- 19126392 TI - Regulation of research through research governance: within and beyond NSW Health. AB - Research governance takes a broad approach to the regulation of human research encompassing: (a) frameworks and systems over ad hoc policy making; (b) quality standards as well as regulatory requirements; and (c) definition of roles and responsibilities of all parties involved in research. The effective and consistent implementation of research governance plays a role in the promotion of quality research. NSW Health has recently issued several policies and procedures in relation to research governance. However, for regimes of research governance to achieve optimal effectiveness, they must be consistent with each other in both the public and private sectors and across Australian jurisdictions. PMID- 19126393 TI - Ensuring safe drinking water in regional NSW: the role of regulation. AB - In regional and rural areas of NSW, drinking water is provided by 107 local water utilities serving a total population of some 1.7 million and operating 323 water supply systems. NSW Health exercises public health oversight of these regional water utilities through the NSW Health Drinking Water Monitoring Program, which provides guidance to water utilities on implementing elements of the Australian Drinking Water Guidelines 2004, including drinking water monitoring. PMID- 19126394 TI - Enforcing law on tobacco sales to minors: getting the question and action right. AB - There is a substantial history of health-related law in Australia, but only recently has this included a significant amount of state regulation pertaining to tobacco promotion, sale and public use. The important question is: under what circumstances do regulation and law enforcement work? Rigorous, energetic, long term local law enforcement on the supply of tobacco to minors demonstrates success in preventing uptake. A model for success combines education, enforcement and publicity; a model used to some effect in alcohol law. Future directions in regulation might include on-the-spot penalties; ensuring the law is simple and has community support; striving for sufficient resources, enthusiasm and skills; and making the tobacco retail industry pay some of the costs of regulating that industry. PMID- 19126395 TI - Public health and regulation of the built environment. AB - The earliest public health statutes contrived to develop effective controls of the excesses of the rapid urban development that accompanied the industrial revolution. By the end of the 20th century, much responsibility for the regulation of the built environment has been dispersed across government and in so doing has moved beyond the gaze of everyday public health concern. This paper argues firstly that there is a strong justification for greater public health involvement in the ongoing regulation of the built environment and secondly that a much broader range of health impacts needs to be considered in the conception of these regulatory measures. PMID- 19126396 TI - Communicable Diseases Report, NSW, September and October 2008. AB - For updated information, including data and facts on specific diseases, visit http://www.health.nsw.gov.au and click on Infectious Diseases or access the site directly at: http://www.health.nsw.gov.au/publichealth/infectious/index.asp. PMID- 19126397 TI - MicroRNA-dependent regulation of cKit in cutaneous melanoma. AB - Loss of cKit receptor in cutaneous melanomas was attributed to the down regulation of AP2 transcription factor. Our analysis of 27 melanoma cell lines showed no correlation between AP2 and c-kit expression. Suggesting a post transcriptional mechanism of cKit down-modulation, we performed genome-wide microRNA (miRNA) expression profiling and found that several miRNA species are commonly up-regulated in melanomas. Among them was mir-221, which can directly interact with c-kit 3'UTR and inhibit cKit protein translation. Observed inverse correlation of the c-kit and mir-221 expression in various melanocytic cells pointed to its involvement in regulation of cKit in melanoma. Moreover, a series of functional assays demonstrated that mir-221 could directly inhibit cKit, p27(Kip1) and, possibly, other pivotal proteins in melanoma. Collectively, the studies presented here indicate that mir-221 could be a novel therapeutic target for the treatment of cutaneous melanoma. They also suggest that regulation of expression and functional activity of identified up-regulated miRNAs should be further studied in the context of malignant melanoma. PMID- 19126398 TI - Aberrant epigenetic reprogramming of imprinted microRNA-127 and Rtl1 in cloned mouse embryos. AB - The microRNA (miRNA) genes mir-127 and mir-136 are located near two CpG islands in the imprinted mouse retrotransposon-like gene Rtl1, a key gene involved in placenta formation. These miRNAs appear to be involved in regulating the imprinting of Rtl1. To obtain insights into the epigenetic reprogramming of cloned embryos, we compared the expression levels of mir-127 and mir-136 in fertilized mouse embryos, parthenotes, androgenotes and cloned embryos developing in vitro. We also examined the DNA methylation status of the promoter regions of Rtl1 and mir-127 in these embryos. Our data showed that mir-127 and mir-136 were highly expressed in parthenotes, but rarely expressed in androgenotes. Interestingly, the expression levels of mir-127 and mir-136 in parthenotes were almost twice that seen in the fertilized embryos, but were much lower in the cloned embryos. The Rtl1 promoter region was hyper-methylated in blastocyst stage parthenotes (75.0%), moderately methylated (32.4%) in the fertilized embryos and methylated to a much lower extent (approximately 10%) in the cloned embryos. Conversely, the promoter region of mir-127 was hypo-methylated in parthenogenetically activated embryos (0.4%), moderately methylated (30.0%) in fertilized embryos and heavily methylated in cloned blastocysts (63-70%). These data support a role for mir-127 and mir-136 in the epigenetic reprogramming of the Rtl1 imprinting process. Analysis of the aberrant epigenetic reprogramming of mir-127 and Rtl1 in cloned embryos may help to explain the nuclear reprogramming procedures that occur in donor cells following somatic cell nuclear transfer (SCNT). PMID- 19126399 TI - Identification and characterization of a leptin-responsive neuroblastoma cell line. AB - The adipocyte-derived hormone leptin plays a critical role in a variety of physiological and pathological actions. As such the determination of leptin signal transduction pathways are important both for understanding the molecular mechanisms of leptin action and for identifying sites for possible therapeutic intervention. Since the hypothalamus is the primary site of leptin action, we sought to identify a neuronal-derived human cell line containing the long form of the leptin receptor (OBRb). To this end, we screened several neuroblastoma cell lines and isolated a sub-line of SH-SY5Y cells, which we designated as SH-OBRb, for further studies. We characterized the transduction pathways induced by leptin in SH-OBRb cells and demonstrated that OBRb mediates tyrosine phosphorylation of STAT3, phosphorylation of ERK1/2, but not SAPK/JNK and p38 MAPK, in a dose and time dependent fashion. In addition, Akt appears to be phosphorylated in the basal state and to be insensitive to further activation by leptin. In summary, we have isolated a unique cell line that can be utilized as a model for use in the study of leptin action and molecular mechanisms. PMID- 19126400 TI - VEGF-A and VEGF-F evoke distinct changes in vascular ultrastructure. AB - VEGF-induced vascular barrier disruptions, formation of fenestrae and vesiculo vacuolar organelles (VVOs), are involved in enhancing vascular permeability in metastatic and edematous diseases. Here, we analyzed vascular permeability enhanced by VEGFs with different receptor selectivity using biological and ultrastructural methods. VEGF-A(165), which stimulates both Flt-1 and KDR, induced the formation of both fenestrae and VVOs at a similar rate, while VEGF-F, a KDR-specific VEGF vammin, predominantly induced the formation of fenestrae with an approximately 5-fold more potent vascular permeability response than VEGF A(165). This is the first report showing that VEGFs with different receptor selectivity evoke distinct changes in vascular ultrastructure. PMID- 19126401 TI - Substrate-specific reduction of PP2A activity exaggerates tau pathology. AB - Phosphorylation of the microtubule-associated protein tau is regulated by the balanced interplay of kinases and phosphatases. Disturbance of this balance causes hyperphosphorylation of tau and neurofibrillary tangle formation in Alzheimer's disease brain. Here, we crossed Dom5 mice that express a substrate specific dominant negative mutant form, L309A Calpha, of protein phosphatase 2A (PP2A) with neurofibrillary-tangle-forming P301L mutant tau transgenic pR5 mice. This exacerbated the tau pathology of pR5 mice significantly. Double-transgenic Dom5/pR5 mice showed 7-fold increased numbers of hippocampal neurons that specifically phosphorylated the pathological S422 epitope of tau. They showed 8 fold increased numbers of tangles compared to pR5 mice, in agreement with our previous finding that tangle formation is correlated with and preceded by phosphorylation of tau at the S422 epitope. This suggests that, in addition to kinases, PP2A and its regulatory subunits may be a therapeutic target for Alzheimer's disease. PMID- 19126402 TI - Human trehalase is a stress responsive protein in Saccharomyces cerevisiae. AB - Three trehalases ATH1, NTH1, and NTH2 have been identified in Saccharomyces cerevisiae. ATH1, and NTH1 hydrolyze trehalose to glucose to provide energy and assist in recovery from stress. Human trehalase (TREH) is expressed in the intestine and kidney and probably hydrolyzes ingested trehalose in the intestine and acts as marker of renal tubular damage in kidney. Since trehalose is not present in circulation or kidney tubules, its renal effect suggests it has other yet unidentified actions. Here we examined the function of human trehalase in budding yeast. We constructed three yeast trehalase mutants (NTH1Delta, NTH2Delta, and ATH1Delta) and then transformed TREH into these mutants. NTH1Delta did not grow on media containing trehalose as the carbon source, and TREH did not rectify NTH1Delta dysfunction and also did not grow on trehalose medium, suggesting that TREH is not responsible for utilization of exogenous trehalose in yeast. In experiments involving exposure to heat, osmotic and oxidative stresses, NTH1Delta showed no recovery. Interestingly, ATH1Delta-TREH showed high sensitivity to all three stressors. ATH1Delta and NTH2Delta showed very low neutral trehalase activity and NTH1Delta did not show any neutral trehalase activity, and trehalose concentrations were higher. Increased neutral trehalase activity (equivalent to the wild type), reduction of trehalose content and brisk sensitivity to stressors were noted in TREH-ATH1Delta strain, but not in TREH NTH1Delta or -NTH2Delta. Our results suggest that TREH acts as a stress-response protein in the kidney rather than involved in utilization of exogenous trehalose. PMID- 19126403 TI - Comparing the mechanical influence of vinculin, focal adhesion kinase and p53 in mouse embryonic fibroblasts. AB - Cytoskeletal reorganization is an ongoing process when cells adhere, move or invade extracellular substrates. The cellular force generation and transmission are determined by the intactness of the actomyosin-(focal adhesion complex) integrin connection. We investigated the intracellular course of action in mouse embryonic fibroblasts deficient in the focal adhesion proteins vinculin and focal adhesion kinase (FAK) and the nuclear matrix protein p53 using magnetic tweezer and nanoparticle tracking techniques. Results show that the lack of these proteins decrease cellular stiffness and affect cell rheological behavior. The decrease in cellular binding strength was higher in FAK- to vinculin-deficient cells, whilst p53-deficient cells showed no effect compared to wildtype cells. The intracellular cytoskeletal activity was lowest in wildtype cells, but increased in the following order when cells lacked FAK+p53>p53>vinculin. In summary, cell mechanical processes are differently affected by the focal adhesion proteins vinculin and FAK than by the nuclear matrix protein, p53. PMID- 19126404 TI - Combined effect of oxidative stress-related gene polymorphisms on atherosclerosis. AB - It is well known that oxidative stress plays critical roles in the pathogenesis of atherosclerosis. In this study, we enrolled 1746 type 2 diabetic subjects, determined 4 common genetic variants related to oxidative stress (glutamate cysteine ligase modifier subunit (GCLM) C-588T, myeloperoxidase G-463A, human paraoxonase 1 Gln192Arg and NAD(P)H oxidase p22phox C242T polymorphisms), and measured carotid intima-media thickness (IMT) as a surrogate marker for atherosclerosis. GCLM C-588T polymorphism was associated with average IMT (AveIMT) (r=0.090, p=0.0008), but the association between the other 3 polymorphisms and AveIMT did not reach the statistical significance. However, AveIMT was significantly greater as the total number of 4 concomitant "pro oxidant alleles" in each subject was increased (r=0.108, p<0.0001). Furthermore, the number of "pro-oxidant alleles" was a risk factor for a high AveIMT independently of conventional risk factors (p=0.0003). In conclusion, accumulation of oxidative stress-associated alleles was associated with carotid atherosclerosis in type 2 diabetic patients. PMID- 19126406 TI - Breast cancer cell response to genistein is conditioned by BRCA1 mutations. AB - Soy phytoestrogens, among which genistein, seem to protect from breast cancer development. In order to study the role of the breast tumour suppressor BRCA1 in response to genistein, we used a new breast cancer cell model: the SUM1315MO2 cell line carrying the 185delAG BRCA1 mutation, which we stably transfected with a plasmid encoding wild-type BRCA1. We showed that growth of BRCA1 mutant cells was strongly inhibited by genistein whereas it only had a weak effect in cells expressing wild-type BRCA1 protein. BRCA1 mutant cells hypersensitivity could be linked to higher expression of ERbeta gene, which suggests that genistein may be an efficient inhibitor of cancer development in BRCA1 mutant breast cancer cells. PMID- 19126405 TI - The actin cytoskeleton differentially regulates NG115-401L cell ryanodine receptor and inositol 1,4,5-trisphosphate receptor induced calcium signaling pathways. AB - Regulation of bi-directional communication between intracellular Ca(2+) pools and surface Ca(2+) channels remains incompletely characterized. We report Ca(2+) release mediated by inositol 1,4,5-trisphosphate receptor (IP(3)R) and ryanodine receptor (RyR) pathways is diminished under actin cytoskeleton disruption in NG115-401L (401L) neuronal cells, yet despite truncated Ca(2+) release, Ca(2+) influx was not significantly altered in these experiments. However, disruption of cortical actin networks completely abolished IP(3)R induced Ca(2+) release, whereas RyR-mediated Ca(2+) release was preserved, albeit attenuated. Moreover, cortical actin disruption completely abolished IP(3)R and RyR linked Ca(2+) influx even though Ca(2+) pool sensitivities were different. These findings suggest discrete Ca(2+) store/Ca(2+) channel coupling mechanisms in the IP(3)R and RyR pathways as revealed by the differential sensitivity to actin perturbation. PMID- 19126407 TI - Decrease of dynamin 2 levels in late-onset Alzheimer's disease alters Abeta metabolism. AB - Late-onset Alzheimer's disease (LOAD) is significantly associated with a single nucleotide polymorphism located in the dynamin (DNM) 2 gene, especially in non carriers of the apolipoprotein E-epsilon4 allele. In this study we used real-time PCR to show that DNM2 mRNA is significantly reduced in the cortex of AD brains and in the peripheral blood of dementia patients. Neuroblastoma cells transfected with a dominant negative DNM2 had increased amyloid beta protein (Abeta) secretion and most of the amyloid precursor protein (APP) in these cells was localized to the plasma membrane. In addition, these cells were rich in flotillin, which is a component of lipid rafts. These data suggest that DNM2 expression is reduced in LOAD, which results in the accumulation of APP in lipid raft-rich plasma membranes. Consequently, Abeta secretion may increase in LOAD neurons. PMID- 19126408 TI - LOX-1 dependent overexpression of immunoglobulins in cardiomyocytes in response to angiotensin II. AB - LOX-1, a cell surface lectin-like receptor, is upregulated by oxidized low density lipoprotein (ox-LDL) and angiotensin II (Ang II), and plays an important role in host defense. The specific C-type lectin domain on LOX-1 is essential for ox-LDL binding and internalization, generation of oxidant species and eliciting immune response. Here, we show that LOX-1 deletion alters genes that relate to immune response. Microarray (and qPCR) analysis of cardiac tissues showed downregulated expression of several immunoglobulins (Igk-V8, Igk-C, Igh-6, Igj, Ighg, Igh, and Igl-V1) in the LOX-1 knockout (KO) mice [p<0.05 vs. the wild-type (WT) mice]. The expression of these immunoglobulins was upregulated several-fold in the LOX-1 KO mice hearts when these mice were infused with Ang II (p<0.05, vs. WT mice). Importantly, cultured mouse HL-1 cardiomyocytes expressed these immunoglobulins, and pretreatment of cardiomyocytes with a specific anti-LOX-1 antibody enhanced the generation of immunoglobulins upon subsequent exposure to Ang II. These observations mirrored the data obtained from WT and LOX-1 KO mice hearts in the resting state and following Ang II infusion. This study provides first set of data on immunoglobulin expression in cardiac tissues of WT and LOX-1 KO mice and in cultured HL-1 cardiomyocytes, and demonstrates that LOX-1 inactivation leads to upregulation of immunoglobulins in cardiomyocytes upon challenge with Ang II. PMID- 19126409 TI - Effects of ice-seeding temperature and intracellular trehalose contents on survival of frozen Saccharomyces cerevisiae cells. AB - Freezing tolerance is an important characteristic for baker's yeast, Saccharomyces cerevisiae, as it is used to make frozen dough. The ability of yeast cells to survive freezing is thought to depend on various factors. The purpose of this work was to study the viability of yeast cells during the freezing process. We examined factors potentially affecting their survival, including the growth phase, ice-seeding temperature, intracellular trehalose content, freezing period, and duration of supercooling. The results showed that the ice-seeding temperature significantly affected cell viability. In the stationary phase, trehalose accumulation did not affect the viability of yeast cells after brief freezing, although it did significantly affect the viability after prolonged freezing. In the log phase, the ice-seeding temperature was more important for cell survival than the presence of trehalose during prolonged freezing. The importance of increasing the extracellular ice-seeding temperature was verified by comparing frozen yeast survival rates in a freezing test with ice seeding temperatures of -5 degrees C and -15 degrees C. We also found that the cell survival rates began to increase at 3h of supercooling. The yeast cells may adapt to subzero temperatures and/or acquire tolerance to freezing stress during the supercooling. PMID- 19126410 TI - Topography of syllable change-detection electrophysiological indices in children and adults with reading disabilities. AB - INTRODUCTION: Developmental dyslexia (DD) is a frequent language-based learning disorder. The predominant etiological view postulates that reading problems originate from a phonological impairment. METHOD: We studied mismatch negativity (MMN) and Late Discriminative Negativity (LDN) to syllables change in both children (n=12; 8-12 years) and young adults (n=15; 14-23 years) with DD compared with controls. RESULTS/DISCUSSION: The present study confirmed abnormal automatic discrimination of syllable changes in both children and adults with developmental dyslexia. MMN topographic, amplitude and latency group differences were evidenced, suggesting different brain mechanisms involved in elementary auditory stimulus change-detection in DD, especially in the left hemisphere. The LDN results demonstrated that the auditory disorder of temporal processing in DD children becomes more serious at late stages of information processing and that the apparent cerebral hypo reactivity to speech changes in DD actually may correspond to additional processes. The age-related differences observed in both MMN and LDN topographies, amplitudes and latency between subjects with DD and controls could indicate different developmental courses in the neural representation of basic speech sounds in good and poor readers, with a tendency to normalization with increasing age. CONCLUSION: Our results showing atypical electrophysiological concomitants of speech auditory perception in DD strongly support the hypothesis of deviant cortical organization in DD. PMID- 19126411 TI - Precision of speed discrimination and smooth pursuit eye movements. AB - Several studies have shown that the precision of smooth pursuit eye speed can match perceptual speed discrimination thresholds during the steady-state phase of pursuit [Kowler, E., & McKee, S. (1987). Sensitivity of smooth eye movement to small differences in target velocity. Vision Research, 27, 993-1015; Gegenfurtner, K., Xing, D., Scott, B., & Hawken, M. (2003). A comparison of pursuit eye movement and perceptual performance in speed discrimination. Journal of Vision, 3, 865-876]. Recently, Osborne et al. [Osborne, L. C., Lisberger, S. G., & Bialek, W. (2005). A sensory source for motor variation. Nature, 437, 412 416; Osborne, L. C., Hohl, S. S., Bialek, W., & Lisberger S. G. (2007). Time course of precision in smooth-pursuit eye movements of monkeys. Journal of Neuroscience, 27, 2987-2998] claimed that pursuit precision during the initiation phase of pursuit also matches the sensory variability, implying that there is no motor noise added during pursuit initiation. However, these results were derived from a comparison of monkey pursuit data to human perceptual data from the literature, which were obtained with different stimuli. To directly compare precision for perception and pursuit, we measured pursuit and perceptual variability in the same human observers using the same stimuli. Subjects had to pursue a Gaussian blob in a step-ramp paradigm and give speed judgments on the same or in different trials. Speed discrimination thresholds were determined for different presentation durations. The analysis of pursuit precision was performed for short intervals containing the initiation period only and also for longer intervals including steady-state pursuit. In agreement with published studies, we found that the Weber fractions for psychophysical speed discrimination were fairly constant for different presentation durations, even for the shortest presentation duration of 150ms. Pursuit variability was 3-4 times as high for the analysis interval (300ms) containing the open-loop phase only. For pursuit analysis intervals of 400-500ms, pursuit variability approached perceptual variability. Our results show that, for the stimuli we used, the motor system contributes at least 50% to the total variability of smooth pursuit eye movements during the initiation phase. PMID- 19126412 TI - Social attachment and aversion in human moral cognition. AB - Modern neuroscience is beginning to substantiate Darwin's notion that the roots of human morality lie in social instincts, present in several species. The role of primitive motivational-emotional systems in human morality still remains under recognized, however. Based on recent experimental evidence and classic neuroanatomical data, we here portray a view of how "ancient" limbic-neurohumoral systems of social attachment and aversion are crucially involved in human moral behaviors, including altruism, empathic concern and aggression. Rather than being a mere evolutionary remnant of our ancestors, such limbic-neurohumoral systems are tightly integrated with cortical mechanisms to enable complex moral sentiments and values, which powerfully influence our choices in socio-cultural settings. Exploring the underlying mechanisms of human social attachment and aversion will provide new insights and foster novel experimental paradigms for the study of moral cognition and behavior. PMID- 19126413 TI - A noninvasive, fast and inexpensive tool for the detection of eye open/closed state in primates. AB - Accurate detection of the eye state (i.e., open or closed) of animals during electrophysiological recordings is often crucial for analyzing physiological data. This requires a system which is reliable, and preferably noninvasive and inexpensive. Here we present such a tool incorporating a standard digital camera and a semi-automatic eye state detection (ESD) algorithm that can be used easily in typical primate electrophysiological setups. The ESD algorithm is based on the high light absorbance of the iris and pupil relative to the eyelid and takes advantage of the unique conditions found in primate physiological recordings (minimal area of sclera and head fixation). The ESD algorithm is as accurate as a human observer, and is not vulnerable to variance inherent to human decisions that it requires (i.e., eye location setting, training set classification and threshold setting). The temporal resolution with standard interlaced digital cameras is 17-20 ms. This is sufficient for the detection of eye state changes during electrophysiological recordings including spontaneous blinking and eye blink conditioning, as demonstrated here. Furthermore, the ESD tool can be applied to other physiological areas of research in which changes in eye state are critical to analyzing neuronal activity. PMID- 19126414 TI - Tumor-necrosis factor-alpha induces retinoic acid-inducible gene-I in rheumatoid fibroblast-like synoviocytes. AB - Tumor-necrosis factor-alpha (TNF-alpha) is a potent proinflammtory cytokine and a key molecule in the pathogenesis of rheumatoid arthritis (RA). Retinoic acid inducible gene-I (RIG-I) is a DExH box protein, which is known to play a role in the inflammatory and immune reactions. We previously reported about potential involvement of RIG-I in synovial inflammation in RA. In the present study, we demonstrated the expression of RIG-I in fibroblast-like synoviocytes stimulated with TNF-alpha. RNA interference against interferon (IFN)-beta abolished the TNF alpha-induced RIG-I expression. In addition, knockdown of RIG-I partially inhibited the TNF-alpha-induced expression of CC chemokine ligand (CCL) 5, a chemokine with chemotactic activity toward lymphocytes and monocytes. These findings suggest that the TNF-alpha/IFN-beta/RIG-I/CCL5 pathway may be involved in the pathogenesis of synovial inflammation in RA. PMID- 19126415 TI - Biological inferences from IgM binding characteristics of recombinant human secretory component mutants. AB - The polymeric immunoglobulin receptor (pIgR) or membrane secretory component (SC) selectively transports polymeric IgA and IgM across secretory epithelial cells to mucosal surfaces. The ligand binding ectodomain consists of five homologous Ig like domains with domain I being an absolute requirement for binding. The role of DII to V in IgM binding remains unknown. Here, using in vitro refolded non glycosylated recombinant domain deletion mutants of human SC, we show by biological and biophysical binding assays that DII to V are required for high affinity binding to IgM. Competitive binding analysis, by whole cell ELISA, showed that DII-V significantly increase the affinity of recombinant SC for IgM (K(i)=2.42 nM) as opposed to recombinant DI only (K(i)=44.8 nM). Lastly, we provide qualitative data highlighting the complexity of measuring the IgM/SC interaction using surface plasmon resonance spectroscopy. PMID- 19126416 TI - Quantitation of HIV-1 group M proviral DNA using TaqMan MGB real-time PCR. AB - The level of human immunodeficiency virus type 1 (HIV-1) proviral DNA is likely to be an important marker of the long-term effectiveness of highly active antiretroviral therapy. A new method was developed for quantifying HIV-1 group M proviral DNA using TaqMan real-time PCR, in which degenerate primers and an MGB probe were used to resolve the difference in amplification efficiencies among different subtypes. The present assay provided good linearity and accuracy in the range of 4-5000 copies of proviral DNA in 0.5microg of cellular DNA. The intra assay and inter-assay coefficients were <31.6% and <30.1%, respectively. In 19 HIV-1 clinical isolates of six subtypes (A, B, C, CRF01_AE, F, and G), quantitation values by the real-time PCR assay matched closely those by Poisson distribution analysis of PCR results at endpoint dilution (R(2)=0.988). This assay is characterized by the use of degenerate primers and having been validated by comparing with a Poisson distribution-based assay. The present real-time PCR assay is highly sensitive, linear, reproducible, accurate, and independent of group M subtypes. The assay will be useful for studying the relationship between HIV-1 proviral loads and the long-term efficacy of antiretroviral therapy for subtype B as well as non-B subtype strains. PMID- 19126417 TI - The role of histamine on cognition. AB - Histamine was intensively studied at the beginning of the 20th century because of its important role in allergic and inflammation processes. In those days it was very difficult that researchers could envisage another impacting function for the imidazolamine in the living systems. Once the imidazolamine was found located in neuron compartment in the brain, increasing evidence supported many regulatory functions including its possible role in memory and learning. The specific participation of histamine in cognitive functions followed a slow and unclear pathway because the many different experimental learning models, pharmacologic approaches, systemic and localized applications of the histamine active compounds into the brain used by researchers showed facilitating or inhibitory effects on learning, generating an active issue that has extended up to present time. In this review, all these aspects are analyzed and discussed considering the many intracellular different mechanisms discovered for histamine, the specific histamine receptors and the compartmentalizing proprieties of the brain that might explain the apparent inconsistent effects of the imidazolamine in learning. In addition, a hypothetical physiologic role for histamine in memory is proposed under the standard theories of learning in experimental animals and humans. PMID- 19126418 TI - Motor coordination of articulators depends on the place of articulation. AB - Although the sounds/p/and/k/are both voiceless plosives, they have different places of articulation: bilabial and velar, respectively. The purpose of this study was to determine the relationship among articulators in plosives with reference to their place of articulation. Ten healthy subjects repeated bilabial and velar plosives in synchronization with magnetic resonance scanning. Each run was measured using a gradient echo sequence. Several linear and angular variables were defined to delineate the individual movements of articulators and to determine the temporal relationships among articulators. These variables showed distinctive changes depending on the place of articulation. In addition, movement of the velum was significantly correlated with that of the lips and the anterior part of the tongue in the bilabial plosive and with the posterior part of the tongue in the velar plosive. We conclude that unitary motor coordination of articulators depends on the place of articulation. PMID- 19126419 TI - FMRFamide-like neuropeptides as putative ligands of the latrophilin-like HC110-R from Haemonchus contortus. AB - The latrophilin-like receptor HC110-R of the parasitic nematode Haemonchus contortus has been previously identified as a target for the novel anthelmintic drug emodepside, but the natural ligand(s) remained completely unknown to date. Here, we investigate 11 different FMRFamide-like neuropeptides as putative ligands by surface plasmon resonance with an immobilized recombinant 54kDa aminoterminal fragment of HC110-R as an interaction partner. AF1, AF10 and PF2 exhibit binding with low affinities as indicated by a K(d) of 11 microM for AF1, 52 microM for AF10 and 583 microM for PF2. Our data indicate that AF1, AF10, and PF2 are putative natural ligands of HC110-R presumably involved in the control of pharyngeal pumping of nematode worms. PMID- 19126420 TI - Elevated pretransplantation soluble CD30 is associated with decreased early allograft function after human lung transplantation. AB - Early allograft function after lung transplantation is variable. Clinical criteria have limited predictive value for early graft function. Recipient immunologic state before LTx may affect early lung function. We investigated the association between pretransplantation soluble CD30 (sCD30), a marker of Th2-type T-cell activation, and early clinical parameters of allograft function. Between September 2002 and January 2007, a total of 80 transplantations were performed at Johns Hopkins Hospital. Of the patients, 43 had a pretransplantation sCD30 level determined. Pre- and postoperative patient variables were collected, and patients were stratified into two groups: sCD30 <20 (low sCD30) and >20 (high sCD30). High sCD30 (n = 26) and low sCD30 (n = 17) groups were similar in age, gender, and ischemia time. In the high sCD30 group, a higher percentage of patients had pulmonary fibrosis and a lower percentage had emphysema. Oxygenation at 48 hours was significantly worse in the high sCD30 group as compared with the low sCD30 (p = 0.003). Moreover, prolonged intubation and 90-day mortality were greater in the high sCD30 group. This represents the first report of the use of sCD30 as a marker for early allograft function in human lung transplanation. Increased pretransplantation recipient sCD30 appears to be associated with decreased early post-transplantation gas exchange, prolonged intubation, and early mortality. PMID- 19126422 TI - The transgenic mouse assay as an alternative test method for regulatory carcinogenicity studies--implications for REACH. AB - REACH, an EU regulation that requires the submission of safety data in support of the protection of human and environmental health, mandates that registration should be achieved with the minimum amount of animal testing possible. Under REACH, a two-year carcinogenicity assay may be required for certain chemicals produced at >1000 metric tonnes per year. In addition, some chemicals that are found to be genotoxic will also require testing. Alternative methods have been explored in an attempt to improve the predictivity of this bioassay as well as to reduce the number of animals used for such testing. This research has focused on the use of transgenic/knockout mouse models. Study results from selected models indicate that they are useful in hazard identification, even if they are not entirely suitable for risk assessment on their own. Carcinogenic hazard assessment can be greatly enhanced and animal use reduced if the traditional two year rat bioassay is combined with a well conducted transgenic mouse assay. Importantly, the use of transgenic animals to supplement a traditional two-year carcinogenicity study may help reduce the number of false negatives, one of the unstated goals of REACH via the precautionary principle. PMID- 19126421 TI - Recombinant attenuated Listeria monocytogenes vaccine expressing Francisella tularensis IglC induces protection in mice against aerosolized Type A F. tularensis. AB - Fransicella tularensis, the causative agent of tularemia, is in the top category (Category A) of potential agents of bioterrorism. To develop a safer vaccine against aerosolized F. tularensis, we have employed an attenuated Listeria monocytogenes, which shares with F. tularensis an intracellular and extraphagosomal lifestyle, as a delivery vehicle for F. tularensis antigens. We constructed recombinant L. monocytogenes (rLm) vaccines stably expressing seven F. tularensis proteins including IglC (rLm/iglC), and tested their immunogenicity and protective efficacy against lethal F. tularensis challenge in mice. Mice immunized intradermally with rLm/iglC developed significant cellular immune responses to F. tularensis IglC as evidenced by lymphocyte proliferation and CD4+ and CD8+ T-cell intracellular expression of interferon gamma. Moreover, mice immunized with rLm/iglC were protected against lethal challenge with F. tularensis LVS administered by the intranasal route, a route chosen to mimic airborne infection, and, most importantly, against aerosol challenge with the highly virulent Type A F. tularensis SchuS4 strain. PMID- 19126423 TI - Tumor promoting and co-carcinogenic effects in medium-term rat hepatocarcinogenesis are not modified by co-administration of 12 pesticides in mixture at acceptable daily intake. AB - The purpose of this investigation was to evaluate the possible influence of a mixture of pesticides on medium-term carcinogenesis using improved hepatocarcinogenesis protocols. We performed a 12 commercially available pesticides combination with alachlor, atrazine, carbofuran, chlorpyrifos, diazinon, dicofol, endosulfan, iprodione, mancozeb, maneb, procymidone and rotenone. The mixture was given at 1-fold and 10-fold the acceptable daily intake (ADI) level in a set of Solt-Farber-derived protocols involving diethylnitrosamine, 2-acetylaminofluorene treatments and a partial hepatectomy. Co-carcinogenic effect and promoting activity were evaluated using gamma-glutamyl transpeptidase (GGT) positive altered hepatocyte foci, as well, protein and mRNA levels of glutathione S-transferase P (GSTP) in liver extracts as molecular biomarkers of carcinogenic effects. The pesticide treatments when compared to vehicle treatments always produced the same number of hepatocyte lesions and an equal GSTP expression on liver extracts independently of carcinogenic-protocol utilized. On this base, we concluded that the pesticide mixture evaluated in this report does not have tumor promoting activity or co-carcinogenic effect in the rat medium-term liver carcinogenesis. Altogether these data contribute to the confidence that the ADI represents a safe intake level to mixture of pesticides at dietary exposure. PMID- 19126424 TI - Cadmium interacts with the transport of essential micronutrients in the mammary gland - a study in rural Bangladeshi women. AB - Although the concentrations of the toxic metal cadmium in breast milk are generally low (< 1 microg/L), experimental studies indicated neurobehavioral and endocrine effects in the suckling offspring. The aim of the present study was to elucidate how cadmium is transported to breast milk by assessing interactions with essential micronutrients. The study is nested into a food and micronutrient supplementation trial conducted among pregnant women in Matlab, a rural area in Bangladesh, where malnutrition is prevalent and the cadmium exposure is relatively high. We measured cadmium in breast milk (BM-Cd; median 0.14 microg/kg; range <0.050-1.0 microg/kg), in erythrocytes (Ery-Cd; median 1.5 microg/kg; range 0.46-4.8 microg/kg) and in urine (U-Cd; median 0.63 microg/L; range 0.050-4.5 microg/L), using inductively coupled plasma mass spectrometry (ICPMS). We found a significant positive association between Ery-Cd and BM-Cd and a breast milk-plasma ratio of approximately 3-4, indicating no barrier against cadmium transport from plasma to breast milk. BM-Cd was positively associated with manganese (r(s)=0.56; p<0.01) and iron (r(s)=0.55; p<0.01) in breast milk, but not with plasma ferritin. On the other hand, BM-Cd was negatively associated with BM-Ca (r(s)=-0.17; p=0.05), indicating that cadmium inhibits the transport of calcium to breast milk. In conclusion, the present study may indicate that cadmium shares common transporters with iron and manganese for transfer to breast milk, but inhibits secretion of calcium to breast milk. PMID- 19126425 TI - Neurochemistry of identified motoneurons of the tensor tympani muscle in rat middle ear. AB - The objective of the present study was to identify efferent and afferent transmitters of motoneurons of the tensor tympani muscle (MoTTM) to gain more insight into the neuronal regulation of the muscle. To identify MoTTM, we injected the fluorescent neuronal tracer Fluoro-Gold (FG) into the muscle after preparation of the middle ear in adult rats. Upon terminal uptake and retrograde neuronal transport, we observed FG in neurons located lateral and ventrolateral to the motor trigeminal nucleus ipsilateral to the injection site. Immunohistochemical studies of these motoneurons showed that apparently all contained choline acetyltransferase, demonstrating their motoneuronal character. Different portions of these cell bodies were immunoreactive to bombesin (33%), cholecystokinin (37%), endorphin (100%), leu-enkephalin (25%) or neuronal nitric oxide synthase (32%). MoTTM containing calcitonin gene-related peptide, tyrosine hydroxylase, substance P, neuropeptide Y or serotonin were not found. While calcitonin gene-related peptide was not detected in the region under study, nerve fibers immunoreactive to tyrosine hydroxylase, substance P, neuropeptide Y or serotonin were observed in close spatial relationship to MoTTM, suggesting that these neurons are under aminergic and neuropeptidergic influence. Our results demonstrating the neurochemistry of motoneuron input and output of the rat tensor tympany muscle may prove useful also for the general understanding of motoneuron function and regulation. PMID- 19126427 TI - In vitro toxic effects of metal compounds on kinetic traits and ultrastructure of rabbit spermatozoa. AB - Metal compounds have been associated with male reproductive toxicity in vivo. The aim of the present study was to investigate the in vitro effects of 20 metal compounds using rabbit ejaculated spermatozoa as a study model for spermiotoxicity. Five of the metals tested (arsenic, cadmium, chromium, mercury and vanadium) reduced sperm motility and curvilinear velocity. Ultrastructural analyses revealed three types of damage to sperm head membranes in relation to the metal used: acrosome breakage with formation of various sized microvesicles (arsenic, cadmium, mercury and platinum); a large round hole (arsenic, cadmium and chromium), and numerous folds in the acrosome membrane (vanadium). The vanadium compound, followed by chromium and mercury compounds, determined a higher number of damaged spermatozoa. In conclusion, all the studied metal compounds, at levels higher than 1microM, may reduce sperm kinetic characteristics and probably fertilizing capacity by triggering specific morphological damages to the head and/or by inhibiting motility. PMID- 19126426 TI - Gonadectomy of male BALB/c mice increases Tim-3(+) alternatively activated M2 macrophages, Tim-3(+) T cells, Th2 cells and Treg in the heart during acute coxsackievirus-induced myocarditis. AB - The incidence of cardiovascular disease, including inflammatory heart diseases like myocarditis, is increased in men. Similarly, male BALB/c mice infected with coxsackievirus B3 (CVB3) develop more severe acute inflammation in the heart compared to females. To better understand the effect of male sex hormones on cardiac inflammation, we gonadectomized (Gdx) male BALB/c mice and examined acute CVB3-induced myocarditis compared to sham controls. Viral replication in the heart was not significantly altered between Gdx and sham mice. However, gonadectomy significantly reduced testosterone levels and inflammation in the heart. FACS analysis of cell populations isolated from the heart revealed that CD11b(+) cells were significantly reduced in Gdx males. However, a GR1(+)F4/80(+) subset of CD11b(+) cells was significantly increased. Because this subset also expressed the interleukin (IL)-4R and IL-10, we refer to these cells as "alternatively activated" or M2 macrophages. A greater percentage of M2 macrophages in Gdx males expressed the inhibitory receptor Tim-3, while fewer expressed IL-1beta and IL-10. Only M2 macrophages upregulated TLR4 and Tim-3, whereas GR1(-)IL-4R(lo) macrophages did not. Additionally, IL-4(+)CD4(+) Th2 cells, Foxp3(+) regulatory T (Treg) cells and Tim-3(+)CD4(+) T cells were significantly increased in the heart following Gdx. Thus, we report for the first time that the inhibitory receptor Tim-3 is expressed on M2 macrophages. Our findings show that sex hormones and/or other mediators released from the testes inhibit anti-inflammatory populations in the heart including Tim-3(+) M2, Tim 3(+)CD4(+) T cells, Th2 and Treg resulting in more severe acute cardiac inflammation in males following CVB3 infection. PMID- 19126428 TI - Alginate-poloxamer microparticles for controlled drug delivery to mucosal tissue. AB - PURPOSE: The aim of this study was to prepare and characterize novel hydrogel based delivery systems allowing for the controlled release of drugs to mucosal surfaces. METHODS: Terbutaline sulfate and bovine serum albumin (BSA)-loaded alginate-poloxamer microparticles were prepared by a w/o-emulsion- and external gelation method. The microparticles were characterized by optical and scanning electron microscopy, laser light diffraction, atomic absorption spectroscopy, energy-dispersive X-ray analysis, via complexation with 1,9-dimethyl methylene blue and using dialysis bags as well as modified Franz diffusion cells for in vitro drug-release measurements. RESULTS: Using heptane as organic phase, homogeneous and almost spherical microparticles were obtained with a high-loading efficiency (>90%). The resulting drug-release patterns could effectively be adjusted by varying the "alginate:poloxamer" blend ratio. In addition, the particle size, morphology, calcium and chloride content as well as alginate release rates could be altered. Erosion was the predominant release mechanism for BSA. Special attention needs to be paid to the microparticle recovery procedure, which can significantly affect key properties such as the resulting drug-release patterns. CONCLUSIONS: The novel hydrogel-based microparticles offering mild conditions for incorporated drugs (e.g., proteins) provide an interesting potential as controlled delivery systems for mucosal surfaces. PMID- 19126429 TI - Novel transdermal delivery of Timolol maleate using sugar esters: preclinical and clinical studies. AB - The feasibility of matrix controlled transdermal patch based on sugar fatty acid ester (SE) as penetration and absorption enhancer containing Timolol maleate (TM) was investigated. The influence of fatty acid type, chain length and hydrophile lipophile balance (HLB) on the in vitro drug release as well as its permeation across hairless rat skin were studied and compared aiming to select a patch formula for clinical performance. Skin irritation induced by SE patch was evaluated by visual scoring, color reflectance measurements and non-invasive transepidermal water loss (TEWL) technique. The results indicated that among different SEs tried, laurate SE with shorter fatty acid chain length and higher HLB value significantly increased the amount of TM liberated from the patch (99+/ 2.1%) and its permeation across rat skin (86+/-4.3%). The total drug permeation and flux values were approximately 5-fold greater compared to SE free patch. The extent of absorption of TM-SE patch expressed by AUC was 64% larger as compared to the oral solution with steady plasma concentration over 18 h and relative bioavailability (F(rel)) of 163%. The developed patch was well tolerated by all the subjects with only moderate skin irritation, which was recovered in 24h after patch removal. The results are very encouraging and offer an alternative approach to maintain higher, prolonged and controlled blood level profile of the drug over 18-24h. PMID- 19126430 TI - Regulation of gene expression by estrogen in mammary gland of wild type and estrogen receptor alpha knockout mice. AB - Using serial analysis of gene expression, we examined the effects of estrogen (E2) replacement in gonadectomized wild type (WT) and E2 receptor alpha knockout (ERalphaKO) mice on global gene expression in mammary gland. In WT mice, a total of 429,302 tags were sequenced, representing the expression level of 99,854 tag species. A total of ten transcripts were found to be modulated by E2, such as sorting nexin 5 and two no match tags. In the ERalphaKO mice, a total of 459,439 tags were sequenced, representing the expression level of 120,149 tag species. Interestingly, the same three transcripts were inversely regulated by E2 in ERalphaKO mice. In total, 78 transcripts were upregulated by E2, while 29 transcripts were downregulated. In contrast to WT mice, the majority of transcripts related to immunity were repressed in ERalphaKO mice. Moreover, induction of transcripts involved in cell differentiation, Ca2+ response, cytoskeleton, protein biosynthesis and secretion, glycolysis, and oxidative phosphorylation were seen only in ERalphaKO mice. The current study will provide useful information to understand the cellular mechanisms of E(2)-mediated gene regulation in tissues in vivo for the development of novel drugs targeting specific ER action in pathological conditions. PMID- 19126431 TI - Transversely isotropic distribution of sulfated glycosaminoglycans in human medial collateral ligament: a quantitative analysis. AB - Decorin and its associated glycosaminoglycan (GAG) side chain, dermatan sulfate (DS), play diverse roles in soft tissue formation and potentially aid in the mechanical integrity of the tissue. Deeper understanding of the distribution and orientation of the GAGs on a microscopic level may help elucidate the structure/function relationship of these important molecules. The hypothesis of the present study was that sulfated GAGs are aligned with transversely isotropic material symmetry in human medial collateral ligament (MCL) with the collagen acting as the axis of symmetry. To test the hypothesis, sulfated GAGs were visualized using transmission electron microscopy (TEM). Three orthogonal anatomical planes were examined to evaluate GAG distributions against symmetry criteria. GAG populations were differentiated using targeted enzyme digestion. Results suggest that sulfated GAGs including DS, chondroitin sulfates A and C, as well as other sub-populations assume transversely isotropic distributions in human MCL. Sulfated GAGs in the plane normal to the collagen axis were found to be isotropic with no preferred orientation. GAGs in the two planes along the collagen axis did not statistically differ and exhibited apparent bimodal distributions, favoring orthogonal distributions with over half at other angles with respect to collagen. A previously developed model, GAGSim3D, was used to interpret potential TEM artifacts. The data collected herein provide refined inputs to micro-scale models of the structure/function relationship of sulfated GAGs in soft tissues. PMID- 19126432 TI - False phylogenies on wood mice due to cryptic cytochrome-b pseudogene. AB - The phylogeny and phylogeography of the Old World wood mice (subgenus Sylvaemus, genus Apodemus, Muridae) are well-documented. Nevertheless, the distributions of species, such as A. fulvipectus and A. ponticus remain dubious, as well as their phylogenetic relationships with A. sylvaticus. We analysed samples of Apodemus spp. across Europe using the mitochondrial cytochrome-b gene (cyt-b) and compared the DNA and amino-acid compositions of previously published sequences. The main result stemming from this study is the presence of a well-differentiated lineage of Sylvaemus including samples of various species (A. sylvaticus, A. fulvipectus, A. ponticus) from distant locations, which were revealed to be nuclear copies of the mitochondrial cyt-b. The presence of this cryptic pseudogene in published sequences is supported by different pathways. This has led to important errors in previous molecular trees and hence to partial misinterpretations in the phylogeny of Apodemus. PMID- 19126433 TI - Inhibition of cyclooxygenase-2 and EP1 receptor antagonism reduces human colonic longitudinal muscle contractility in vitro. AB - We investigated the contribution of cyclo-oxygenase enzyme inhibition and prostamide agonism on human colonic contractility in vitro. The effects of the non-specific COX inhibitor diclofenac were compared against selective COX-2 inhibition via nimesulide, the prostanoid EP(1) receptor antagonist SC19220 or the prostaglandin prodrug/prostamide receptor agonist bimatoprost, on potency of contraction to acetylcholine in human colonic circular and longitudinal muscle strips. Pre-treatment with either nimesulide (10(-5)M) or diclofenac (10(-6)M) caused a significant decrease in the potency of acetylcholine-evoked longitudinal muscle contraction, but did not inhibit acetylcholine-evoked circular muscle contraction. Pre-treatment with the EP(1) receptor antagonist SC19220 (10(-5)M) similarly decreased cholinergic potency in longitudinal muscle, without influence on circular muscle contraction. The prostamide agonist bimatoprost (10(-6)M) increased basal circular and longitudinal muscle tone, but did not alter cholinergic potency in either muscle layer. In conclusion, colonic longitudinal muscle contraction is augmented by COX-2 activity, most likely via PGE(2) acting at EP(1) receptors. While colonic contraction is tonically modulated by bimatoprost, it does not share the same functional properties attributed to other endogenous COX-2 metabolites on colonic contractile function. PMID- 19126434 TI - Biology of endocannabinoid synthesis system. AB - Endocannabinoids (endogenous ligands of cannabinoid receptors) exert diverse physiological and pathophysiological functions in animal tissues. N Arachidonoylethanolamine (anandamide) and 2-arachidonoylglycerol (2-AG) are two representative endocannabinoids. Both the compounds are arachidonic acid containing lipid molecules generated from membrane glycerophospholipids, but their biosynthetic pathways are totally different. Anandamide is principally formed together with other N-acylethanolamines (NAEs) in a two-step pathway, which is composed of Ca(2+)-dependent N-acyltransferase and N acylphosphatidylethanolamine-hydrolyzing phospholipase D (NAPE-PLD). cDNA cloning of NAPE-PLD and subsequent analysis of its gene-disrupted mice led to the discovery of alternative pathways comprising multiple enzymes. As for the 2-AG biosynthesis, recent results, including cDNA cloning of diacylglycerol lipase and analyses of phospholipase Cbeta-deficient mice, demonstrated that these two enzymes are responsible for the in vivo formation of 2-AG functioning as a retrograde messenger in synapses. In this review article, we will focus on recent progress in the studies on the enzymes responsible for the endocannabinoid biosyntheses. PMID- 19126435 TI - Shrinkage of the hippocampal remnant after surgery for temporal lobe epilepsy: impact on seizure and neuropsychological outcomes. AB - The aim of this study was to investigate the influence of the postoperative hippocampal remnant on postoperative seizure and neuropsychological outcome in temporal lobe epilepsy (TLE). Postoperative volumetric MRI measurements of 53 patients surgically treated for TLE revealed a postoperative volume loss of the hippocampal remnant compared with the respective preoperative segment in all patients. Extent of preoperative hippocampal pathology, remnant shrinkage, resection volume, and postoperative volume of the hippocampal remnant did not correlate with seizure outcome 1 year after surgery. With respect to neuropsychological outcome, performance on tasks assessing verbal memory and language-related functions was impaired in patients with left-sided pathology after surgery. Performance of patients with right-sided pathology (n=26) demonstrated no significant correlation with hippocampal measures or with neuropsychological data. Degree of hippocampal remnant shrinkage seems to be associated with decreased verbal memory performance in patients with left-sided TLE. PMID- 19126436 TI - Association of antiepileptic drug nonadherence with risk of seizures in adults with epilepsy. AB - This study evaluated the potential effect of antiepileptic drug (AED) nonadherence on the risk of subsequent seizure. Retrospective insurance claims from the United States were analyzed. Inclusion criteria were: age 21-64 years, diagnosis of epilepsy or nonfebrile convulsions, 2 AED prescriptions, and insurance enrollment for 6 months pre- and 60 days post-AED initiation. Seizure was defined as a hospital or emergency admission associated with epilepsy or nonfebrile convulsions. Observation began 7 days post-drug initiation, ending with the first of the following: seizure, insurance disenrollment, or 365 days post-drug initiation. Adherence was measured using the medication possession ratio (MPR), with MPR <0.8 defining nonadherence. Seizure risk was assessed using an extended Cox proportional hazards model. Of 18,073 subjects identified, 2467 (14%) had 1 seizure. Mean follow-up was 133 days among subjects with event and 305 days for patients without event. Seizure risk was 21% higher among nonadherers (hazard ratio=1.205, P=0.0002) than adherers. PMID- 19126437 TI - Quality of life in childhood epilepsy: what is the level of agreement between youth and their parents? AB - Children and parents evaluate the child's quality of life (QOL) from their own perspectives; therefore, responses may differ, especially in abstract domains. We examined differences between self- and proxy-reported QOL of children with epilepsy. Children with active epilepsy (N=375) and their parents (N=378) separately completed the CHEQOL-25, a condition-specific QOL measure. The intraclass correlation coefficient was used to determine interrater agreement. Concordance on the Total CHEQOL-25 was 0.45 (P<0.01). Discrepancies were greatest for the subscales of Secrecy (0.24, P<0.01) and Present Concerns (0.32, P<0.01). School placement correlated with discrepancy in the Intrapersonal/Emotional subscale (r=0.19, P<0.05), and the child's age at testing correlated with discrepancy of the Total measure (r=0.15, P<0.01). This study demonstrates that parent perspectives alone are insufficient to measure their child's QOL. The CHEQOL-25 is a practical tool, with complementary parent and child versions, which can be used to determine health-related quality of life in children with epilepsy. PMID- 19126438 TI - Self-appraisal and objective assessment of cognitive and affective functioning in persons with epileptic and nonepileptic seizures. AB - Subjective and objective assessments of cognitive and affective functioning of patients admitted to the EMU with epileptic (ES, n=22) and psychogenic nonepileptic (NES, n=23) seizures were compared. It was hypothesized that the patients with NES would overestimate their cognitive impairments and underestimate their affective disturbances relative to those with ES. Results revealed that the patients with NES reported greater word-finding difficulty than those with ES (P=0.02), but performed better than the patients with ES on the Boston Naming Test (BNT, P=0.03), suggesting a tendency to overestimate word finding difficulty. Patients with NES and ES did not otherwise differ in subjective ratings of cognitive functions or emotional state; however, patients with NES performed worse on a test of affect expression/perception compared with those with ES (P=0.02). For patients with NES, only performance on memory tests was significantly correlated with their anxiety level. Obtaining both subjective ratings and objective test findings of cognitive and affective functioning may help further differentiate between patients with NES and those with ES. PMID- 19126439 TI - Symptomatic complex partial status epilepticus manifesting as utilization behavior of a mobile phone. AB - Utilization behavior (UB) consists of reaching out and using objects in the environment in an automatic manner and out of context. This behavior has been correlated to frontal lobe dysfunction, especially of the right hemisphere. We describe a 60-year-old woman, affected by a glioblastoma located in the right frontal region, who presented with intermittent UB of the mobile phone as the main clinical manifestation of partial complex status epilepticus. Video/EEG studies showed a striking correlation between mobile phone utilization and ictal epileptic activity. Clinical and EEG findings were markedly reduced after the introduction of antiepileptic drugs. This case study suggests that UB may be added to the symptoms described for partial seizures originating from frontal areas. PMID- 19126440 TI - Neonatal status epilepticus alters prefrontal-striatal circuitry and enhances methamphetamine-induced behavioral sensitization in adolescence. AB - Neonatal seizures may alter the developing neurocircuitry and cause behavioral abnormalities in adulthood. We found that rats previously subjected to lithium pilocarpine (LiPC)-induced neonatal status epilepticus (NeoSE) exhibited enhanced behavioral sensitization to methamphetamine (MA) in adolescence. Neurochemically, dopamine (DA) and metabolites were markedly decreased in prefrontal cortex (PFC) and insignificantly changed in striatum by NeoSE, but were increased in both PFC and striatum by NeoSE+MA. Glutamate levels were increased in both PFC and striatum in the NeoSE+MA group. DA turnover, an index of utilization and activity, was increased by NeoSE but reversed by MA in PFC. Gene expression of the regulator of G-protein signaling 4 (RGS4) was downregulated in PFC and striatum by NeoSE and further suppressed by MA. These findings suggest NeoSE affects both dopaminergic and glutamatergic systems in the prefrontal-striatal circuitry that manifests as enhanced behavioral sensitization to MA in adolescence. PMID- 19126441 TI - Multiple myeloma causing jaundice and duodenal ulceration. PMID- 19126442 TI - CD209 gene polymorphisms in South Indian HIV and HIV-TB patients. AB - Dendritic-cell-specific intercellular adhesion molecule-3 (ICAM-3)-grabbing non integrin (DC-SIGN), a pattern recognition receptor, is associated with immune functions and is also exploited by HIV-1 and Mycobacterium tuberculosis as a part of their immune evasion strategy. In the present study we investigated whether variants in the DC-SIGN encoding CD209 gene are associated with susceptibility to or protection against HIV-1 infection as well as development of tuberculosis (TB) among HIV-1 infected south Indian patients. CD209 gene variants in the promoter region (-336 and -139), in the intron and 3'-untranslated regions (In2+11 and 2281) were studied using polymerase chain reaction-based genotyping methods in 131 HIV patients without TB (HIV+TB-) and 107 HIV patients with TB (HIV+TB+), 107 HIV negative pulmonary TB patients (HIV-PTB+) and 157 healthy controls. Results revealed a decreased frequency of -336 G/G genotype among all HIV patients compared to healthy controls and -336 G/G genotype was not observed among HIV+TB- individuals (p=0.005; odds ratio (OR) 0 (95% confidence intervals (CI) 0-0.46); Peto's odds ratio 0.149 (95% CI 0.045-0.50)). Among HIV+ patients, those with TB had a significantly increased frequency of -336 G/G genotype (p=0.003; OR undefined; Peto's odds ratio 9.8 (95% CI 2.2-44.3)) compared to those without TB. Other polymorphisms were not significantly different between the various study groups. The results suggest that -336 G/G genotype while associated with protection against HIV-1 infection the same genotype is also associated with susceptibility to HIV-TB among south Indians. PMID- 19126443 TI - Tuberculoma of the cavernous sinus mimicking a meningioma: case report and review of the literature. AB - Cavernous sinus involvement of tuberculosis is rare; only seven cases have been reported in the literature to date. Many factors make the diagnosis difficult in this location, necessitating a surgical procedure. We report the case of a 45 year-old man who presented with a left cavernous sinus syndrome clinically. The lesion mimicked a meningioma of the cavernous sinus in radiological examinations. It was entirely excised through a subtemporal approach. Histopathological examination revealed caseating granuloma and the diagnosis of tuberculoma was established. Postoperative antituberculous drugs completed the treatment and led to complete resolution of neurological symptoms. In our paper, we emphasize the rarity of the lesion in this location, we review all the cases previously published underlining the main epidemiological, clinical, radiological, therapeutic and prognostic features. PMID- 19126444 TI - Circadian variation of heart rate turbulence and the number of ventricular premature beats in patients with mitral valve prolapse. PMID- 19126445 TI - Calorimetric studies of the effect of cis-carotenoids on the thermotropic phase behavior of phosphatidylcholine bilayers. AB - Carotenoid geometry is a factor that determines their solubility and orientation in the lipid membrane as well as antioxidant capacities and bioavailability. The effects of the cis-isomers of carotenoids (zeaxanthin and beta-carotene) on the thermotropic properties of lipid membranes formed with dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) were investigated by means of differential scanning calorimetry. The results were compared with the effects caused by the all-trans-isomer. Both the trans and cis isomers of zeaxanthin shifted the main phase transition temperature to lower values and decreased the cooperativity of the phase transition. The effect of all trans zeaxanthin on the physical properties of the lipid bilayers has been shown to strongly depend on the hydrocarbon chain length of the membrane. In the case of cis-zeaxanthin this relationship is weaker. PMID- 19126446 TI - Studies on the interaction between imidacloprid and human serum albumin: spectroscopic approach. AB - The interaction between imidacloprid (IMI) and human serum albumin (HSA) was investigated using fluorescence and UV/vis absorption spectroscopy. The experimental results showed that the fluorescence quenching of HSA by IMI was a result of the formation of IMI-HSA complex; static quenching was confirmed to result in the fluorescence quenching. The apparent binding constant K(A) between IMI and HSA at three differences were obtained to be 1.51 x 10(4), 1.58 x 10(4), and 2.19 x 10(4) L mol(-1), respectively. The thermodynamic parameters, DeltaH degrees and DeltaS degrees were estimated to be 28.44 kJ mol(-1), 174.76 J mol( 1) K(-1) according to the van't Hoff equation. Hydrophobic interactions played a major role in stabilizing the complex. The distance r between donor (HSA) and acceptor (IMI) was obtained according to fluorescence resonance energy transfer. The effect of IMI on the conformation of HSA was analyzed using synchronous fluorescence spectroscopy CD and three-dimensional fluorescence spectra, the environment around Trp and Tyr residues were altered. PMID- 19126447 TI - A minimal model of tumor growth inhibition. AB - The preclinical development of antitumor drugs greatly benefits from the availability of models capable of predicting tumor growth as a function of the drug administration schedule. For being of practical use, such models should be simple enough to be identifiable from standard experiments conducted on animals. In the present paper, a stochastic model is derived from a set of minimal assumptions formulated at cellular level. Tumor cells are divided in two groups: proliferating and nonproliferating. The probability that a proliferating cell generates a new cell is a function of the tumor weight. The probability that a proliferating cell becomes nonproliferating is a function of the plasma drug concentration. The time-to-death of a nonproliferating cell is a random variable whose distribution reflects the nondeterministic delay between drug action and cell death. The evolution of the expected value of tumor weight obeys two differential equations (an ordinary and a partial differential one), whereas the variance is negligible. Therefore, the tumor growth dynamics can be well approximated by the deterministic evolution of its expected value. The tumor growth inhibition model, which is a lumped parameter model that in the last few years has been successfully applied to several antitumor drugs, is shown to be a special case of the minimal model presented here. PMID- 19126448 TI - Physiologically based pharmacokinetic (PBPK) models for ethanol. AB - Physiologically based pharmacokinetic models have been used to describe the distribution and elimination of ethanol after intravenous administration. These models have been used to estimate the ethanol infusion profile that is sufficient for achieving a prescribed breath ethanol concentration time course in individuals, providing a useful platform for several pharmacokinetic and pharmacodynamic investigations. Mathematical foundations of these models are examined, including the derivation of an explicit set of governing equations in the form of a system of nonlinear ordinary differential equations. These equations can then be used to formulate and refine parameter identification and control strategies. Finally, a framework in which models related to this model can be constructed and analyzed is described. PMID- 19126449 TI - Efficient fully implicit time integration methods for modeling cardiac dynamics. AB - Implicit methods are well known to have greater stability than explicit methods for stiff systems, but they often are not used in practice due to perceived computational complexity. This paper applies the backward Euler (BE) method and a second-order one-step two-stage composite backward differentiation formula (C BDF2) for the monodomain equations arising from mathematically modeling the electrical activity of the heart. The C-BDF2 scheme is an L-stable implicit time integration method and easily implementable. It uses the simplest forward Euler and BE methods as fundamental building blocks. The nonlinear system resulting from application of the BE method for the monodomain equations is solved for the first time by a nonlinear elimination method, which eliminates local and nonsymmetric components by using a Jacobian-free Newton solver, called Newton- Krylov solver. Unlike other fully implicit methods proposed for the monodomain equations in the literature, the Jacobian of the global system after the nonlinear elimination has much smaller size, is symmetric and possibly positive definite, which can be solved efficiently by standard optimal solvers. Numerical results are presented demonstrating that the C-BDF2 scheme can yield accurate results with less CPU times than explicit methods for both a single patch and spatially extended domains. PMID- 19126450 TI - Model-based detection of heart rate turbulence. AB - In this study, the integral pulse frequency modulation model is extended to account for the presence of ectopic beats and heart rate turbulence (HRT). Based on this model, a new statistical approach to the detection and characterization of HRT is presented. The detector structure involves a set of Karhunen-LoEve basis functions and a generalized likelihood ratio test statistic T(x) . The three most significant basis functions reflect the difference in heart rate prior to a ventricular ectopic beat (VEB) compared to after HRT, the "average" HRT, and a delayed contribution to HRT, respectively. Detector performance was studied on both simulated and ECG signals. Three different simulations were performed for the purpose of studying the influence of SNR, QRS jitter, and ECG sampling rate. The results show that the HRT test statistic T(x) performs better in all simulations than do the commonly used parameters known as turbulence onset (TO) and turbulence slope (TS). In order to attain the same performance as T(x), TS needs at least twice the amount of VEBs for averaging, and TO at least four times. The detector performance was also studied on ECGs acquired from eight patients who underwent hemodialysis treatment with the goal to discriminate between patients considered to be hypotension-resistant (HtR) and hypotension prone (HtP). The results show that T(x) exhibits larger mean values in HtR patients than in HtP, suggesting that HRT is mostly present in HtR patients. The overlap between the two groups was larger for TO and TS than for T(x). PMID- 19126451 TI - Improved time--frequency analysis of atrial fibrillation signals using spectral modeling. AB - In patients with atrial fibrillation (AF), the fibrillatory frequency trend and the time-dependent spectral characteristics can be investigated using a spectral profile technique. The spectral profile is updated by fitting each short-time spectrum. The aim of this study is to develop model-based means for stricter control on the update of the spectral profile. A spectral model defined by a superposition of Gaussian functions is suggested for describing the fundamental and harmonics of the atrial waves during AF, thereby accounting for basic characteristics of the typical AF spectrum. The model parameters are obtained from weighted least squares fitting of the model to the observed spectrum. The method was tested on simulated signals as well as on 48 ECG recordings from 15 patients with persistent AF. Using simulated signals, we assessed the accuracy in terms of magnitude and width of the spectral peaks. For SNR=0 dB, the maximum normalized error was less than 0.2 when estimating magnitude of both the fundamental and the harmonics, whereas it was less than 0.15 for the fundamental and 0.7 for the harmonics with respect to the estimation of the width. We observed a marked improvement while tracking the main fibrillatory frequency as the error was reduced by more than 50% in comparison with the original method. Analyzing ECGs, reliable spectral profiles were obtained in all recordings, even in those cases (5/48) that were not well characterized by the original method. PMID- 19126452 TI - Human hip joint cartilage: MRI quantitative thickness and volume measurements discriminating acetabulum and femoral head. AB - This paper aims at developing a quantitative system for measuring human hip cartilage thickness and volume using magnetic resonance imaging (MRI). A new MRI acquisition technique, named axial rotation, where the acquisition planes are organized around a virtual axis, was used. The MRI protocol consists of a 2-D multiple-echo data image combination (MEDIC) using water excitation. Inner and outer interface contours of acetabulum and femoral head cartilage are obtained using a semiautomated 3-D segmentation method and combined to form 3-D surfaces. A local spherical coordinate system computed from the original contours enables cartilage thickness and volume computation. An anatomical labeling is performed automatically for thickness and volume measurements in predefined subregions: inferior, anterior, superior, and posterior. A registration module is introduced allowing the assessment of cartilage changes over time. Validation of the system was conducted with three protocols each involving data obtained from nine subjects: 1) registration process accuracy; 2) intrareader reproducibility; and 3) intervisit coefficient of variation. Data showed excellent correlation coefficients for either the intrareader (r>or=0.0942, p<0.0001 ) or intervisit (r>or=0.0837, p<0.005) protocols. This noninvasive system, which enables the quantification of cartilage thickness and volume in the human hip joint using MRI, is the first to discriminate the acetabular and femoral head cartilage throughout the entire hip without the use of an external device, and to implement hip registration for follow-up studies on the same subject. PMID- 19126453 TI - Adaptive and robust methods of reconstruction (ARMOR) for thermoacoustic tomography. AB - In this paper, we present new adaptive and robust methods of reconstruction (ARMOR) for thermoacoustic tomography (TAT), and study their performances for breast cancer detection. TAT is an emerging medical imaging technique that combines the merits of high contrast due to electromagnetic or laser stimulation and high resolution offered by thermal acoustic imaging. The current image reconstruction methods used for TAT, such as the delay-and-sum (DAS) approach, are data-independent and suffer from low-resolution, high sidelobe levels, and poor interference rejection capabilities. The data-adaptive ARMOR can have much better resolution and much better interference rejection capabilities than their data-independent counterparts. By allowing certain uncertainties, ARMOR can be used to mitigate the amplitude and phase distortion problems encountered in TAT. The excellent performance of ARMOR is demonstrated using both simulated and experimentally measured data. PMID- 19126454 TI - Effect of erythrocyte aggregation on hematocrit measurement using spectral-domain optical coherence tomography. AB - Optical coherence tomography (OCT) has the potential to be a noninvasive method for hematocrit (HCT) measurement. This study shows the effect of erythrocyte aggregation at the level seen in healthy humans and pathological states on HCT measurement based on monitoring the slope changes in OCT depth reflectivity profile using spectral-domain optical coherence tomography (SDOCT). Our measurement indicates that the HCT estimated by SDOCT depends on the erythrocyte aggregation state and the flow rate. Measured HCT in blood samples with 0.6% and 2% dextran 500 is underestimated by 4.5%-10.5% and 17.1%-19.5% for HCT from 35% 55% at a flow rate of 4.7 mm/s. Underestimation is smaller at a high flow rate as compared to a low flow rate, indicating erythrocyte aggregation is an important factor that will affect accurate HCT measurement using SDOCT. PMID- 19126455 TI - A legged anchoring mechanism for capsule endoscopes using micropatterned adhesives. AB - This paper presents a new concept for an anchoring mechanism to enhance existing capsule endoscopes. The mechanism consists of three actuated legs with compliant feet lined with micropillar adhesives to be pressed into the intestine wall to anchor the device at a fixed location. These adhesive systems are inspired by gecko and beetle foot hairs. Single-leg and full capsule mathematical models of the forces generated by the legs are analyzed to understand capsule performance. Empirical friction models for the interaction of the adhesives with an intestinal substrate were experimentally determined in vitro using dry and oil-coated elastomer micropillar arrays with 140 microm pillar diameter, 105 microm spacing between pillars, and an aspect ratio of 1:1 on fresh porcine small intestine specimens. Capsule prototypes were also tested in a simulated intestine environment and compared with predicted peristaltic loads to assess the viability of the proposed design. The experimental results showed that a deployed 10 gr capsule robot can withstand axial peristaltic loads and anchor reliably when actuation forces are greater than 0.27 N using dry micropillars. Required actuation forces may be reduced significantly by using micropillars coated with a thin silicone oil layer. PMID- 19126456 TI - Taser blunt probe dart-to-heart distance causing ventricular fibrillation in pigs. AB - The maximum distance between the heart and a model Taser stimulation dart, called the dart-to-heart distance, at which the Taser can directly cause ventricular fibrillation (VF), was measured in pigs. A 9-mm-long blunt probe was advanced snugly through the surrounding tissues toward the heart. Five animals [pig mass=61.2+/-6.23 standard deviation (SD) kg] for ten dart-to-heart distances where the Taser caused VF were tested. The dart-to-heart distances where the Taser caused VF of the first stimulation site ranged from 4 to 8 mm with average 6.2 mm+/-1.79 (SD) and of the second stimulation site ranged from 2 to 8 mm with average 5.4 mm+/-2.41 (SD). The results help inform the evolving discussion of risks associated with Tasers. PMID- 19126457 TI - 3-D-microwave breast tumor detection: study of system performance. AB - This paper presents a study of performance for a 3-D breast tumor detection system. The system is based on processing the scattered signals when a narrow pulse is transmitted from a set of antennas placed surrounding the breast. The 3 D system performance is evaluated by placing the tumor at any breast position, even at locations near the breast muscle where detection is more difficult. Results are obtained for different sizes and positions of the abnormalities that are defined as spherical and bunch-shaped geometries, which approach the well known breast tumors morphology. These results show that the array distribution considered will have a high impact on detection. PMID- 19126458 TI - Improved detectability in medical microwave radio-thermometers as obtained by active antennas. AB - Microwave radiometry is a spectral measurement technique for resolving blackbody radiation of heated matter above absolute zero. The emission levels vary with frequency and are at body temperatures maximized in the infrared spectral band. Medical radio-thermometers are mostly noninvasive short-range instruments that can provide temperature distributions in subcutaneous biological tissues when operated in the microwave region. However, a crucial limitation of the microwave radiometric observation principle is the extremely weak signal level of the thermal noise emitted by the lossy material (-174 dBm/Hz at normal body temperature). To improve the radiometer SNR, we propose to integrate a tiny, moderate gain, low-noise preamplifier (LNA) close to the antenna terminals as to obtain increased detectability of deep seated thermal gradients within the volume under investigation. The concept is verified experimentally in a lossy phantom medium by scanning an active antenna across a thermostatically controlled water phantom with a hot object embedded at 38 mm depth. Three different setups were investigated with decreasing temperature contrasts between the target and ambient medium. As a direct consequence of less ripple on the raw radiometric signal, statistical analysis shows a marked increase in signal-to-clutter ratio of the brightness temperature spatial scan profiles, when comparing active antenna operation with conventional passive setups. PMID- 19126459 TI - A device for noninvasive assessment of vascular impairment risk in the lower extremity. AB - The repeatability and resolution of the clinical gold standard of vascular assessment, the ankle-brachial index (ABI), was compared to that of a new device that dynamically assesses tissue perfusion during external loading utilizing laser Doppler flowmetry. Eight subjects of varying levels of vascular impairment were tested in successive weeks using two different sites on the subject's posterior calf. These new measures included the perfusion decrease as well as the unloading delay during cyclic loading. Some new dynamic tissue perfusion measures demonstrated comparable levels of reproducibility with the ABI (e.g., 10%-20%). Only the unloading delay showed potentially enhanced resolution over ABI measures. The perfusion decrease showed little resolution, and the remaining parameters exhibited too great variability (25%-90%). The unloading delay associated with the reperfusion response during cyclic loading displayed the greatest combination of reproducibility and differentiation between subject groups of varying levels of vascular impairment. The preliminary results of this pilot study were also used to estimate sample sizes necessary to detect possible significant (P<0.05) differences between subject groups for all measured perfusion parameters. From these calculations, at least 30 subjects are needed for future study in each of the five subject groups. PMID- 19126461 TI - Tumor response estimation in radar-based microwave breast cancer detection. AB - Radar-based microwave imaging techniques have been proposed for early stage breast cancer detection. A considerable challenge for the successful implementation of these techniques is the reduction of clutter, or components of the signal originating from objects other than the tumor. In particular, the reduction of clutter from the late-time scattered fields is required in order to detect small (subcentimeter diameter) tumors. In this paper, a method to estimate the tumor response contained in the late-time scattered fields is presented. The method uses a parametric function to model the tumor response. A maximum a posteriori estimation approach is used to evaluate the optimal values for the estimates of the parameters. A pattern classification technique is then used to validate the estimation. The ability of the algorithm to estimate a tumor response is demonstrated by using both experimental and simulated data obtained with a tissue sensing adaptive radar system. PMID- 19126460 TI - Development of anatomically realistic numerical breast phantoms with accurate dielectric properties for modeling microwave interactions with the human breast. AB - Computational electromagnetics models of microwave interactions with the human breast serve as an invaluable tool for exploring the feasibility of new technologies and improving design concepts related to microwave breast cancer detection and treatment. In this paper, we report the development of a collection of anatomically realistic 3-D numerical breast phantoms of varying shape, size, and radiographic density which can readily be used in finite-difference time domain computational electromagnetics models. The phantoms are derived from T1 weighted MRIs of prone patients. Each MRI is transformed into a uniform grid of dielectric properties using several steps. First, the structure of each phantom is identified by applying image processing techniques to the MRI. Next, the voxel intensities of the MRI are converted to frequency-dependent and tissue-dependent dielectric properties of normal breast tissues via a piecewise-linear map. The dielectric properties of normal breast tissue are taken from the recently completed large-scale experimental study of normal breast tissue dielectric properties conducted by the Universities of Wisconsin and Calgary. The comprehensive collection of numerical phantoms is made available to the scientific community through an online repository. PMID- 19126462 TI - Evaluation of corneal refraction in a model of a gaze tracking system. AB - The complexity of corneal refraction modeling complicates geometrical analysis of gaze tracking systems. The task can be accomplished in different ways by means of different approximations. Depending on the assumptions made, errors can arise and the accuracy of trackers can end up determined to an excessive degree by the hardware setup and subject parameters. In this study, keeping the hardware and eye model the same, we compare different methods for evaluation of corneal refraction. Our results show that the approximated methods are sensitive to the calibration of the system and less robust to noise. The nonapproximated method removes any dependency on setup and allows clear differentiation between hardware and subject's calibration. PMID- 19126463 TI - Orthogonal field calibration analysis for myocardial electrode arrays used in defibrillation studies. AB - Mapping the myocardial electric field during a defibrillation pulse requires the recording of potential differences between electrodes. The field is then calculated from these quantities and the corresponding calibration matrix. One straightforward calibration technique involves alignment of a known electric field along each of the orthogonal axes of an electrode array and recording the resulting potential differences. However, no results have been reported to support the efficacy of this technique. This study performs a detailed error analysis including a one-to-one comparison to a precision calibration technique, and quantitatively establishes the efficacy of the orthogonal field technique. PMID- 19126464 TI - A novel 3-D image-based morphological method for phenotypic analysis. AB - A new approach for the study of geometric morphometrics is presented based on well-established image processing techniques in a novel combination to support high-throughput analysis necessary for large-scale determination of genotype phenotype relationships. The method retains full 3-D data, and avoids manual landmark selection. Micro-computed tomography images are superimposed into a common orientation by rigid image registration with an isotropic scale factor. An average sample shape is determined by averaging the intensities of corresponding voxels of the registered images, and shape variation is determined by calculating the image gradient of the average shape. Localized shape differences between mean images or between an individual and a group mean are identified and quantified by surface-to-surface distance measures of superimposed images. Validation was performed using geometric shapes of known dimensions as well as biological samples of C57 BL/6 J and A/WySnJ mouse skulls, and shape variation of the mouse skulls was consistent with previously published results. Although the image gradient is sensitive to both image registration and filtration of the average image, the effect can be minimized by consistent use of image analysis parameters. While the proposed approach deviates from well-established landmark based geometric morphometric tools, it is not intended to replace these current methods. Rather, it will be an important contribution to provide high-throughput screening in large-scale studies focused on understanding genotype-phenotype relationships so that subsequent morphometric approaches using established techniques can be better focused. PMID- 19126465 TI - Effects of audio compression in automatic detection of voice pathologies. AB - This paper investigates the performance of an automatic system for voice pathology detection when the voice samples have been compressed in MP3 format and different binary rates (160, 96, 64, 48, 24, and 8 kb/s). The detectors employ cepstral and noise measurements, along with their derivatives, to characterize the voice signals. The classification is performed using Gaussian mixtures models and support vector machines. The results between the different proposed detectors are compared by means of detector error tradeoff (DET) and receiver operating characteristic (ROC) curves, concluding that there are no significant differences in the performance of the detector when the binary rates of the compressed data are above 64 kb/s. This has useful applications in telemedicine, reducing the storage space of voice recordings or transmitting them over narrow-band communications channels. PMID- 19126466 TI - A novel method for characterization of peripheral nerve fiber size distributions by group delay. AB - The ability to determine the characteristics of peripheral nerve fiber size distributions would provide additional information to clinicians for the diagnosis of specific pathologies of the peripheral nervous system. Investigation of these conditions, using electrodiagnostic techniques, is advantageous in the sense that such techniques tend to be minimally invasive yet provide valuable diagnostic information. One of the principal electrodiagnostic tools available to the clinician is the nerve conduction velocity test. While the peripheral nerve conduction velocity test can provide useful information to the clinician regarding the viability of the nerve under study, it is a single-parameter test that yields no detailed information about the characteristics of the functioning nerve fibers within the nerve trunk. In this study, we present a technique based on decomposition of the maximal compound evoked potential and subsequent determination of the group delay of the contributing nerve fibers. The fiber group delay is then utilized as an initial estimation of the nerve fiber size distribution and the associated temporal propagation delays of the single-fiber evoked potentials to a reference electrode. Simulation studies, based on deterministic single-fiber action potential functions, are used to demonstrate the robustness of the proposed technique in the presence of simulated noise associated with the recording process. PMID- 19126467 TI - Measuring saccade peak velocity using a low-frequency sampling rate of 50 Hz. AB - During the last decades, small head-mounted video eye trackers have been developed in order to record eye movements. Real-time systems-with a low sampling frequency of 50/60 Hz-are used for clinical vestibular practice, but are generally considered not to be suited for measuring fast eye movements. In this paper, it is shown that saccadic eye movements, having an amplitude of at least 5 degrees, can, in good approximation, be considered to be bandwidth limited up to a frequency of 25-30 Hz. Using the Nyquist theorem to reconstruct saccadic eye movement signals at higher temporal resolutions, it is shown that accurate values for saccade peak velocities, recorded at 50 Hz, can be obtained, but saccade peak accelerations and decelerations cannot. In conclusion, video eye trackers sampling at 50/60 Hz are appropriate for detecting the clinical relevant saccade peak velocities in contrast to what has been stated up till now. PMID- 19126468 TI - Comments on "Sliding mode closed-loop control of FES: controlling the shank movement". AB - There are some essential problems with the arguments presented in the above paper about the design of a sliding-mode controller for functional electrical simulation (FES) induced control of knee-joint angle. In this note, we show that applying some approximations in derivation of the control law violates the reaching condition and could introduce some parasitic unmodeled dynamics in the sliding-mode control loop. Therefore, the proposed controller cannot force the system into a sliding-mode regime, and its ability of producing a robust control loop with good tracking performance is theoretically under question. PMID- 19126469 TI - Speckle suppression in SAR images using the 2-D GARCH model. AB - A novel Bayesian-based speckle suppression method for Synthetic Aperture Radar ( SAR) images is presented that preserves the structural features and textural information of the scene. First, the logarithmic transform of the original image is analyzed into the multiscale wavelet domain. We show that the wavelet coefficients of SAR images have significantly non-Gaussian statistics that are best described by the 2-D GARCH model. By using the 2-D GARCH model on the wavelet coefficients, we are capable of taking into account important characteristics of wavelet coefficients, such as heavy tailed marginal distribution and the dependencies between the coefficients. Furthermore, we use a maximum a posteriori (MAP) estimator for estimating the clean image wavelet coefficients. Finally, we compare our proposed method with various speckle suppression methods applied on synthetic and actual SAR images and we verify the performance improvement in utilizing the new strategy. PMID- 19126470 TI - Ubiquitously supervised subspace learning. AB - In this paper, our contributions to the subspace learning problem are two-fold. We first justify that most popular subspace learning algorithms, unsupervised or supervised, can be unitedly explained as instances of a ubiquitously supervised prototype. They all essentially minimize the intraclass compactness and at the same time maximize the interclass separability, yet with specialized labeling approaches, such as ground truth, self-labeling, neighborhood propagation, and local subspace approximation. Then, enlightened by this ubiquitously supervised philosophy, we present two categories of novel algorithms for subspace learning, namely, misalignment-robust and semi-supervised subspace learning. The first category is tailored to computer vision applications for improving algorithmic robustness to image misalignments, including image translation, rotation and scaling. The second category naturally integrates the label information from both ground truth and other approaches for unsupervised algorithms. Extensive face recognition experiments on the CMU PIE and FRGC ver1.0 databases demonstrate that the misalignment-robust version algorithms consistently bring encouraging accuracy improvements over the counterparts without considering image misalignments, and also show the advantages of semi-supervised subspace learning over only supervised or unsupervised scheme. PMID- 19126471 TI - Analytic estimation of subsample spatial shift using the phases of multidimensional analytic signals. AB - In this correspondence, a method of analytic subsample spatial shift estimation based on an a priori n-D signal model is proposed. The estimation uses the linear phases of n analytic signals defined with the multidimensional Hilbert transform. This estimation proposes: i) an analytic solution to the n-D shift estimation and ii) an estimation without processing complex cross-correlation function or cross spectra between signals contrary to most phase shift estimators. The method provides better performance in estimating subsample shifts than two classical estimators, one using the maximum of cross-correlation function and the other seeking the zero of the complex correlation function phase. Two delay estimators using the in-phase and quadrature-phase components of signals are also compared to our estimator. Like most estimators using the complex signal phases, the estimator proposed herein presents the advantage of unaltered accuracy when low sampled signals are used. Moreover, we show that this method can be applied to motion tracking with ultrasound images. Thus, included in a block-based motion estimation method and tested with ultrasound data, this estimator provides an analytical solution to the translation estimation problem. PMID- 19126472 TI - Error protection and interleaving for wireless transmission of JPEG 2000 images and video. AB - The transmission of JPEG 2000 images or video over wireless channels has to cope with the high probability and burstyness of errors introduced by Gaussian noise, linear distortions, and fading. At the receiver side, there is distortion due to the compression performed at the sender side, and to the errors introduced in the data stream by the channel. Progressive source coding can also be successfully exploited to protect different portions of the data stream with different channel code rates, based upon the relative importance that each portion has on the reconstructed image. Unequal Error Protection (UEP) schemes are generally adopted, which offer a close to the optimal solution. In this paper, we present a dichotomic technique for searching the optimal UEP strategy, which lends ideas from existing algorithms, for the transmission of JPEG 2000 images and video over a wireless channel. Moreover, we also adopt a method of virtual interleaving to be used for the transmission of high bit rate streams over packet loss channels, guaranteeing a large PSNR advantage over a plain transmission scheme. These two protection strategies can also be combined to maximize the error correction capabilities. PMID- 19126474 TI - Markerless human motion analysis in Gauss-Laguerre transform domain: an application to sit-to-stand in young and elderly people. AB - A markerless computer vision technique specifically designed to track natural elements on the human body surface is presented. The method implements the estimate of translation, rotation, and scaling by means of a maximum likelihood approach carried out in the Gauss-Laguerre transform domain. The approach is particularly suitable for human movement analysis in clinical contexts, where kinematics is at present performed by means of marker-based systems. Specific drawbacks of these latter systems, such as the burden of time for marker placement and the intrinsic intrusive nature, would be removed by the proposed method. Experimental results in terms of tracking performance are obtained by analyzing video sequences capturing the execution of the sit-to-stand task in two groups of young and elderly volunteers. The results are compared with clinical studies that used marker-based systems, and are particularly encouraging for a future extension of the approach to other motor tasks and to predict scores obtained from the physical performance batteries that are widely and regularly used by clinicians and physical therapists. PMID- 19126473 TI - Sensorless motion planning for medical needle insertion in deformable tissues. AB - Minimally invasive medical procedures such as biopsies, anesthesia drug injections, and brachytherapy cancer treatments require inserting a needle to a specific target inside soft tissues. This is difficult because needle insertion displaces and deforms the surrounding soft tissues causing the target to move during the procedure. To facilitate physician training and preoperative planning for these procedures, we develop a needle insertion motion planning system based on an interactive simulation of needle insertion in deformable tissues and numerical optimization to reduce placement error. We describe a 2-D physically based, dynamic simulation of needle insertion that uses a finite-element model of deformable soft tissues and models needle cutting and frictional forces along the needle shaft. The simulation offers guarantees on simulation stability for mesh modifications and achieves interactive, real-time performance on a standard PC. Using texture mapping, the simulation provides visualization comparable to ultrasound images that the physician would see during the procedure. We use the simulation as a component of a sensorless planning algorithm that uses numerical optimization to compute needle insertion offsets that compensate for tissue deformations. We apply the method to radioactive seed implantation during permanent seed prostate brachytherapy to minimize seed placement error. PMID- 19126475 TI - An extended Markov blanket approach to proteomic biomarker detection from high resolution mass spectrometry data. AB - High-resolution matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has recently shown promise as a screening tool for detecting discriminatory peptide/protein patterns. The major computational obstacle in finding such patterns is the large number of mass/charge peaks (features, biomarkers, data points) in a spectrum. To tackle this problem, we have developed methods for data preprocessing and biomarker selection. The preprocessing consists of binning, baseline correction, and normalization. An algorithm, extended Markov blanket, is developed for biomarker detection, which combines redundant feature removal and discriminant feature selection. The biomarker selection couples with support vector machine to achieve sample prediction from high-resolution proteomic profiles. Our algorithm is applied to recurrent ovarian cancer study that contains platinum-sensitive and platinum-resistant samples after treatment. Experiments show that the proposed method performs better than other feature selection algorithms. In particular, our algorithm yields good performance in terms of both sensitivity and specificity as compared to other methods. PMID- 19126478 TI - Formation of barium strontium titanate thin films via electrophoretic deposition process. AB - Synthesis of crystalline barium stronium titanate (Ba(0.6)Sr(0.4)TiO(3)) nanoparticles and subsequent formation of thin films have been carried out. The crystalline products were confirmed by X-ray diffractometry. Uniform Ba(0.6)Sr(0.4)TiO(3) thin films were formed by using electrophoretic deposition method (EPD) under a 0.3 to 5 V dc bias for 10 min to 1 h. Ba(0.6)Sr(0.4)TiO(3) nanoparticles having an average crystallite size of 20 to 50 nm, and Ba(0.6)Sr(0.4)TiO(3) thin films with thickness of 150 nm to 4 mum were obtained. A scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to characterize the morphologies of nanoparticles and thin films. The results show that the EPD process route is a rapid, cost-effective alternative for forming Ba(0.6)Sr(0.4)TiO(3) thin films. PMID- 19126479 TI - A six-DOF prismatic-spherical-spherical parallel compliant nanopositioner. AB - A nanopositioner using a 6-prismatic-spherical-spherical parallel (PSS) linked compliant mechanism driven by 6 multilayered piezoelectric actuators (PZT) is presented. Compared with a traditional Gough-Stewart platform in which each actuator was installed between the end effector and the base, this nanopositioner installed the PZT directly on the base to achieve much smaller mechanical loop, higher stiffness, faster response, and compactness. This nanopositioner consists of one fixed plate; three 2-PSS compliant mechanisms; and one end effector. The kinematics characteristics of the nanopositioner were analyzed through the pseudo rigid-body model. The behavior of the compliant mechanism was intensively simulated by the finite element method (FEM). Tracking a 5 nm radius circle of the 15 x 15 x 5 cm(3) prototype was experimentally demonstrated. The measurement results showed the nanopositioner achieved 8 microm travel with 5 nm resolutions and 200 microrad rotation with 0.7 microrad resolutions. The nanopositioner can be used to manipulate nano scale devices, fabricate nano components, or operate nano machines. PMID- 19126480 TI - Performance of thin-film ferroelectric capacitors for EMC decoupling. AB - This paper studied the effects of thin-film ferroelectrics as decoupling capacitors for electromagnetic compatibility applications. The impedance and insertion loss of PZT capacitors were measured and compared with the results from commercial off-the-shelf capacitors. An equivalent circuit model was extracted from the experimental results, and a considerable series resistance was found to exist in ferroelectric capacitors. This resistance gives rise to the observed performance difference around series resonance between ferroelectric PZT capacitors and normal capacitors. Measurements on paraelectric (Ba,Sr)TiO(3) based integrated varactors do not show this significant resistance. Some analyses were made to investigate the mechanisms, and it was found that it can be due to the hysteresis in the ferroelectric thin films. PMID- 19126481 TI - Effects of electric field and biaxial flexure on the failure of poled lead zirconate titanate. AB - Reliable design of lead zirconate titanate (PZT) piezo stack actuators demands that several issues, including electromechanical coupling and ceramic strength size scaling, be scrutinized. This study addresses those through the use of ball on-ring (BoR) biaxial flexure strength tests of a PZT piezoelectric material that is concurrently subjected to an electric field. The Weibull strength distributions and fracture surfaces were examined. The mechanical failures were further analyzed in terms of internal stress, energy release rate, and domain switching toughening. Both the sign and the magnitude of an electric field had a significant effect on the strength of poled PZT within the tested range. A surface flaw type with an average depth of around 18 microm was identified to be the strength limiter and responsible for the failure of the tested PZT under both mechanical and electromechanical loadings. With a value of 0.74 MPa.m(1/2) in the absence of electric field, the fracture toughness of the poled PZT was affected by an applied electric field just as the strength was affected. These results and observations have the potential to serve probabilistic reliability analysis and design optimization of multilayer PZT piezo actuators. PMID- 19126482 TI - Self-sensing force control of a piezoelectric actuator. AB - This paper describes an approach to controlling the force generated by a piezoelectric actuator (PEA) accurately without using any force sensor. A model PEA is proposed that includes a new asymmetric hysteresis operator and that takes the external force into account. A detection model is deduced that allows computation in real time of the PEA elongation and generated force starting from the measurement of the driving voltage and current. This detection model is used to replace a force sensor for closed-loop force control of a PEA. Experiments are carried out using 2 PEAs in an experimental setup. The first actuator is the controlled actuator, and the second one is used as a dynamic controllable mechanical load. It is shown that a good control performance can be obtained whatever the mechanical loading conditions. PMID- 19126483 TI - Microwave dielectric properties of Na(x)Nd((2-x)/3)TiO(3) solid solutions. AB - Na(x)Nd((2-x)/3)TiO(3) solid solutions possess tetragonal or orthorhombic perovskite structure, where the A-sites are characterized by 2 kinds of cations (Na(+), Nd(3+)) and vacancies. We have measured microwave dielectric properties of Na(x)Nd((2-x)/3)TiO(3) solid solutions (x = 0.05 to 0.5) to investigate an effect of the compositional ordering in the A-sites of the perovskite structure. According to powder x-ray diffraction, the A-site is disordered in the composition range of x = 0.29 to 0.5. A compositional ordering (Na(+), vacancy / Nd(3+)) in A-sites appeared when x = 0.05 to 0.2. The quality factor (Q x f), where Q is the inverse of dielectric loss and f is frequency, was found to be slightly improved with decreasing Na content in the range of x = 0.05 to 0.2, suggesting that the Q x f of the Na(x)Nd((2-x)/3)TiO(3) solid solutions depends on the compositional ordering in A-sites. PMID- 19126484 TI - A compensated multi-pole linear ion trap mercury frequency standard for ultra stable timekeeping. AB - The multi-pole linear ion trap frequency standard (LITS) being developed at the Jet Propulsion Laboratory (JPL) has demonstrated excellent short- and long-term stability. The technology has now demonstrated long-term field operation providing a new capability for timekeeping standards. Recently implemented enhancements have resulted in a record line Q of 5 x 10(12) for a room temperature microwave atomic transition and a short-term fractional frequency stability of 5 x 10(-14)/tau(1/2). A scheme for compensating the second order Doppler shift has led to a reduction of the combined sensitivity to the primary LITS systematic effects below 5 x 10(-17) fractional frequency. Initial comparisons to JPL's cesium fountain clock show a systematic floor of less than 2 x 10(-16). The compensated multi-pole LITS at JPL was operated continuously and unattended for a 9-mo period from October 2006 to July 2007. During that time it was used as the frequency reference for the JPL geodetic receiver known as JPLT, enabling comparisons to any clock used as a reference for an International GNSS Service (IGS) site. Comparisons with the laser-cooled primary frequency standards that reported to the Bureau International des Poids et Mesures (BIPM) over this period show a frequency deviation less than 2.7 x 10(-17)/day. In the capacity of a stand-alone ultra-stable flywheel, such a standard could be invaluable for long term timekeeping applications in metrology labs while its methodology and robustness make it ideal for space applications as well. PMID- 19126485 TI - Thin-film piezoelectric-on-silicon resonators for high-frequency reference oscillator applications. AB - This paper studies the application of lateral bulk acoustic thin-film piezoelectric-on-substrate (TPoS) resonators in high-frequency reference oscillators. Low-motional-impedance TPoS resonators are designed and fabricated in 2 classes--high-order and coupled-array. Devices of each class are used to assemble reference oscillators and the performance characteristics of the oscillators are measured and discussed. Since the motional impedance of these devices is small, the transimpedance amplifier (TIA) in the oscillator loop can be reduced to a single transistor and 3 resistors, a format that is very power efficient. The lowest reported power consumption is approximately 350 microW for an oscillator operating at approximately 106 MHz. A passive temperature compensation method is also utilized by including the buried oxide layer of the silicon-on-insulator (SOI) substrate in the structural resonant body of the device, and a very small (-2.4 ppm/ degrees C) temperature coefficient of frequency is obtained for an 82-MHz oscillator. PMID- 19126486 TI - Effective lateral modulations with applications to shear modulus reconstruction using displacement vector measurement. AB - High accuracy in measuring target motions can be realized by combined use of our previously developed lateral Gaussian envelope cosine modulation method (LGECMM) and displacement vector measurement methods that enable simultaneous axial and lateral displacement measurements, such as the multidimensional autocorrelation method (MAM). In this paper, LGECMM is improved by using parabolic functions and Hanning windows instead of Gaussian functions in the apodization function, i.e., parabolic apodization and Hanning apodization. The new modulations enable decreases in effective aperture length (i.e., channels) and yield more accurate displacement vector measurements than LGECMM due to increased echo signal-to noise ratio and lateral spatial resolution. That is, on the basis of a priori knowledge about ultrasound propagation using the focusing scheme and shape of the apodization function, we stopped using Fraunhofer approximation. As practical applications of the modulations, for an agar phantom that is deformed in a lateral direction, stable and accurate 2-D shear modulus reconstructions are performed using our previously developed direct inversion approach together with 2-D strain tensor measurements using MAM. PMID- 19126487 TI - Simultaneous imaging of artery-wall strain and blood flow by high frame rate acquisition of RF signals. AB - Mechanical properties of the arterial walls are significantly altered by atherosclerosis, and various studies have been recently conducted to measure the regional elastic properties (radial strain) of the arterial wall. We have developed a phase-sensitive correlation-based method, namely, the phased-tracking method, to measure the regional radial strain. On the other hand, the measurement of blood flow is an important practical routine in the diagnosis of atherosclerosis. It would be useful if the regional strain of the arterial wall as well as blood flow could be assessed simultaneously. Such measurement would require a high frame rate of several kilohertz. In this study, acquisition of ultrasonic RF echoes at a high frame rate (about 3500 Hz) was achieved using parallel beamforming in which plane waves were transmitted only 3 times and receive beamforming created 24 beams for each transmit beam. The accuracy in measurement of the minute radial strain was evaluated by a basic experiment using a cylindrical phantom. The error of the measured strain from the theoretical strain profile and its standard deviation were 4.8% and 9.5%, respectively. Furthermore, the radial strain of a carotid arterial wall and blood flow were simultaneously imaged in vivo. PMID- 19126488 TI - Time-delay estimation in ultrasound echo signals using individual sample tracking. AB - The performance of many signal-processing applications in ultrasound medical imaging, including elastography, blood flow imaging, tissue velocity imaging, and acoustic radiation force impulse imaging, depends on the performance of their time-delay estimators. In this paper, we present a new time-delay estimator based on the tracking of individual samples using a continuous representation of the echo signal. We also present the use of the same interpolation approach to improve the performance of the zero-crossing tracking delay estimator. Simulation results show that sample tracking algorithms significantly outperform commonly used window-based algorithms in terms of bias and standard deviation. Sample tracking algorithms also have higher sensitivity and resolution compared with traditional delay estimators, including recently introduced spline-based continuous time-delay estimators, because they provide the displacement of individual samples. Experimental results demonstrating the viability of sample tracking delay estimation are also presented. PMID- 19126490 TI - Quantitative modeling of reflected ultrasonic bounded beams and a new estimate of the Schoch shift. AB - The wavefields of bounded acoustic beams and pulses reflected from water-loaded plates are fully modeled with the phase advance technique. The wavefield produced at the source is propagated at any incidence angle using phase shift modeling that incorporates the full analytic solution for the acoustic reflectivity at the interface. This approach provides for the ready visualization of both the stationary monofrequency beam wavefield and animation of the temporally bounded pulse. The model images are reminiscent of the classic Schlieren photographs that first illustrated the nonspecular behavior of the reflected beams incident near critical angles. Various phenomena such as the lateral displacement and the null zone at the Rayleigh critical angle are recreated. A new approximation for this shift agrees well with that of the peak energy of the reflected beam. Similar effects are observed during the reflection of a bounded pulse. Although more computationally costly than existing analytic approximations, the phase advance technique can facilitate the interpretation of reflectivity measurements obtained in laboratory experiments. In particular, the full visualization allows for a better understanding of the behavior of reflected waves at any angle of incidence. PMID- 19126489 TI - Forward-looking intracardiac ultrasound imaging using a 1-D CMUT array integrated with custom front-end electronics. AB - Minimally invasive catheter-based electrophysiological (EP) interventions are becoming a standard procedure in diagnosis and treatment of cardiac arrhythmias. As a result of technological advances that enable small feature sizes and a high level of integration, nonfluoroscopic intracardiac echocardiography (ICE) imaging catheters are attracting increasing attention. ICE catheters improve EP procedural guidance while reducing the undesirable use of fluoroscopy, which is currently the common catheter guidance method. Phased-array ICE catheters have been in use for several years now, although only for side-looking imaging. We are developing a forward-looking ICE catheter for improved visualization. In this effort, we fabricate a 24-element, fine-pitch 1-D array of capacitive micromachined ultrasonic transducers (CMUT), with a total footprint of 1.73 mm x 1.27 mm. We also design a custom integrated circuit (IC) composed of 24 identical blocks of transmit/ receive circuitry, measuring 2.1 mm x 2.1 mm. The transmit circuitry is capable of delivering 25-V unipolar pulses, and the receive circuitry includes a transimpedance preamplifier followed by an output buffer. The CMUT array and the custom IC are designed to be mounted at the tip of a 10-Fr catheter for high-frame-rate forward-looking intracardiac imaging. Through-wafer vias incorporated in the CMUT array provide access to individual array elements from the back side of the array. We successfully flip-chip bond a CMUT array to the custom IC with 100% yield. We coat the device with a layer of polydimethylsiloxane (PDMS) to electrically isolate the device for imaging in water and tissue. The pulse-echo in water from a total plane reflector has a center frequency of 9.2 MHz with a 96% fractional bandwidth. Finally, we demonstrate the imaging capability of the integrated device on commercial phantoms and on a beating ex vivo rabbit heart (Langendorff model) using a commercial ultrasound imaging system. PMID- 19126491 TI - A comparison of laser ultrasound measurements and finite-element simulations for the dispersion behavior of antisymmetric flexural modes propagating along wedge tips with coatings. AB - Antisymmetric flexural (ASF) modes are antisymmetric types of guided waves propagating along the tip of wedge-shaped waveguides. Acoustic sensors frequently rely on the detection of small mass changes that result from binding a coated layer coupled to the active sensor surface. While a layer is coated on one of the wedge's surfaces, another type of sensor can be potentially developed based on detecting the change of ASF velocity. This paper describes a study on the effects of ASF dispersion behavior for a wedge with a layer of coating using a combined numerical and experimental investigation. In this study, the frequency range is from 0.5 MHz to 10 MHz, and the effective wave propagation length along the wedge tip ranges from 3 mm to 13 mm. Brass wedge tips coated with aluminum layer are studied for the case of slow matrix with fast coating, while aluminum wedge tips with copper coatings are studied for the case of fast matrix/slow coating combination. Like surface acoustic waves propagating along a flat surface with a layer of coating, loaded and stiffened phenomena are observed for the ASF modes traveling along coated wedges. Moreover, the wedge tip geometry is found to have an effect in enhancing the loaded and stiffened phenomena. The numerical results show good agreement with experimental results. PMID- 19126492 TI - 20 MHz/40 MHz dual element transducers for high frequency harmonic imaging. AB - Concentric annular type dual element transducers for second harmonic imaging at 20 MHz / 40 MHz were designed and fabricated to improve spatial resolution and depth of penetration for ophthalmic imaging applications. The outer ring element was designed to transmit the 20 MHz signal and the inner circular element was designed to receive the 40 MHz second harmonic signal. Lithium niobate (LiNbO(3)), with its low dielectric constant, was used as the piezoelectric material to achieve good electrical impedance matching. Double matching layers and conductive backing were used and optimized by KLM modeling to achieve high sensitivity and wide bandwidth for harmonic imaging and superior time-domain characteristics. Prototype transducers were fabricated and evaluated quantitatively and clinically. The average measured center frequency for the transmit ring element was 21 MHz and the one-way --3 dB bandwidth was greater than 50%. The 40 MHz receive element functioned at 31 MHz center frequency with acceptable bandwidth to receive attenuated and frequency downshifted harmonic signal. The lateral beam profile for the 20 MHz ring elements at the focus matched the Field II simulated results well, and the effect of outer ring diameter was also examined. Images of a posterior segment of an excised pig eye and a choroidal nevus of human eye were obtained both for single element and dual element transducers and compared to demonstrate the advantages of dual element harmonic imaging. PMID- 19126493 TI - Modeling the radiation of ultrasonic phased-array transducers with Gaussian beams. AB - A new transducer beam model, called a multi-Gaussian array beam model, is developed to simulate the wave fields radiated by ultrasonic phased-array transducers. This new model overcomes the restrictions on using ordinary multi Gaussian beam models developed for large single-element transducers in phased array applications. It is demonstrated that this new beam model can effectively model the steered and focused beams of a linear phased-array transducer. PMID- 19126494 TI - High-temperature piezoelectric single crystal ReCa(4)O(BO(3))(3) for sensor applications. AB - Large-size and high-quality ReCa(4)O(BO(3))(3) (ReCOB, Re = rare earth) single crystals were grown by the Czochralski pulling method. In this work, the electrical properties were investigated at room temperature and elevated temperature for YCa(4)O(BO(3))(3) (YCOB). The dielectric permittivity, piezoelectric strain coefficient, and electromechanical coupling were found to be on the order of 11, 6.5 pC/N, and 12.5%, respectively, with a high piezoelectric voltage coefficient around 0.067 Vm/N. The electrical resistivity of YCOB was found to be 2 x 10(8) Ohm.m at 800 degrees C, with Q values of 4,500 at 950 degrees C. The frequency/temperature coefficient of YCOB was found to be -75 to 85ppm/K in the temperature range of 30 to 950 degrees C, depending on the crystal orientations. Together with their temperature-independent properties, ReCOB crystals are promising candidates for sensing applications at elevated temperatures. PMID- 19126495 TI - Propagation characteristics of shear horizontal surface acoustic waves in (11 2 0) ZnO film/silica glass substrate structures. PMID- 19126496 TI - Characterization of a 40-MHz focused transducer with a fiber grating laser hydrophone. PMID- 19126497 TI - An integrated optoacoustic transducer combining etalon and black PDMS structures. PMID- 19126498 TI - Theoretical and experimental study of plate acoustic waves in ZX-cut lithium niobate. PMID- 19126499 TI - Influence of extrinsic contribution on the macroscopic properties of hard and soft lead zirconate titanate ceramics. PMID- 19126500 TI - Primary ovarian pregnancy. PMID- 19126501 TI - The roles of endoscopy in endometrial cancer. AB - Advancements in technology have increased the availability of valuable minimally invasive techniques for gynecologists to use in the management of patients with endometrial cancer. Hysteroscopy has recently been confirmed as an accurate diagnostic method for endometrial carcinoma. Several retrospective studies have found increased positive peritoneal cytology in women who underwent hysteroscopy, but recent studies have indicated that there is currently no evidence to suggest that diagnostic hysteroscopy increases the risk of malignant cells spreading into the peritoneal cavity, or worsens the prognosis in women with endometrial carcinoma. Laparoscopy plays an important role in treating endometrial cancer. Most studies have shown no differences in recurrence rates or survival between patients who underwent laparoscopic or abdominal staging surgery. Laparoscopy offers many advantages, including avoidance of an abdominal incision, a shorter hospital stay, and a probable more rapid recovery time. PMID- 19126502 TI - Differential diagnosis of gynecologic organ-related diseases in women presenting with ascites. AB - Ascites is a pathologic accumulation of fluid within the peritoneal cavity, and usually develops as a result of liver disease, congestive heart failure or nephrotic syndrome. Ascites is also a common manifestation of some gynecologic diseases. It is important that health care workers consider gynecologic problems among the potential differential diagnoses in patients presenting with ascites. Various kinds of ovarian diseases, such as epithelial ovarian cancer, benign ovarian fibroma, stromal hyperplasia, ovarian hyperstimulation syndrome, primary peritoneal serous carcinoma, endometriosis and peritoneal tuberculosis, should be kept in mind when women are found to have ascites. PMID- 19126503 TI - Meconium peritonitis in utero---the value of prenatal diagnosis in determining neonatal outcome. AB - OBJECTIVE: Meconium peritonitis (MP) is a chemical peritonitis caused by fetal intestinal perforation in utero. Its incidence is rare, but serious neonatal morbidity or even mortality can occur if the diagnosis is not made until after birth. Prenatal diagnosis is important in prompting early postnatal surgical intervention, and so improving neonatal outcome. MATERIALS AND METHODS: Fourteen cases diagnosed in utero with MP from January 1996 to December 2005 were enrolled in this study. The final diagnoses were established by surgical findings or postnatal radiography. The prenatal ultrasound features, neonatal birth characteristics, surgical findings, postnatal management and neonatal outcomes were reviewed. RESULTS: All infants received follow-up care at our hospital. Prenatal ultrasound findings included fetal ascites (100%), intra-abdominal calcification (93%), dilated bowel loops (57%), pseudocysts (29%), and polyhydramnios (50%). Four infants (4/14; 28.5%) did not undergo postnatal surgery, but survived well. The mean gestational age at detection was significantly earlier in the non-surgery group (23+/-3.6 weeks) than in the surgery group (31.7+/-2.5 weeks). One infant (7.1%) died because of sepsis after two neonatal operations. The overall survival rate was 92.9%. CONCLUSION: MP can be diagnosed by prenatal ultrasound, and the neonatal outcome is favorable. Early detection is not associated with poor neonatal outcome, and selective termination is unnecessary. Resolution of dilated bowel loops and polyhydramnios predict a low rate of postnatal surgical intervention. PMID- 19126504 TI - Fetal acidosis from obstetric interventions during the first vaginal delivery. AB - OBJECTIVE: The aim of this study was to analyze the blood gas values of umbilical cord blood in newborns of first vaginal deliveries with or without obstetric interventions. MATERIALS AND METHODS: In a prospective descriptive study conducted during the 6-month period from August 2003 through February 2004 at a university hospital, we analyzed the umbilical cord blood gas results of 80 term newborns delivered vaginally from healthy nulliparous women. Multivariate logistic analysis was used to evaluate the associations between fetal acidosis (pH<7.20) and any obstetric interventions. RESULTS: The mean of umbilical cord blood arterial pH was 7.26 (standard deviation, 0.072). After controlling for the confounding factors with multivariate logistic regression, fetal acidosis (pH<7.20) was found to be significantly associated with oxytocin augmentation (odds ratio [OR], 16.48; 95% confidence interval [CI], 1.21-226.1) and vacuum extraction (OR, 10.76; 95% CI, 1.025-112.9). In contrast, there was no significant relationship between fetal acidosis with episiotomy (OR, 1.096; 95% CI, 0.07-16.6) or epidural anesthesia (OR, 0.074; 95% CI, 0.003-2.09). CONCLUSION: Oxytocin augmentation and vacuum extraction were significantly related to low cord arterial pH values (pH<7.20), but there were no adverse effects to the newborns of first vaginal deliveries. PMID- 19126505 TI - Predictive factors and short-term fetal outcomes of breech presentation: a case control study. AB - OBJECTIVE: This study evaluated the predictive factors and short-term fetal outcomes of breech presentation by comparing breech and cephalic pregnancies of >or=36 weeks gestation. MATERIALS AND METHODS: Two hundred and one breech and 149 cephalic pregnancies of >or=36 weeks gestation, with no other maternal or fetal problems, were compared with regard to placental localization, fetal heart rate variability, smoking, body mass index, maternal weight gain, placental weight, birth weight, sex, Apgar scores, and umbilical cord length. RESULTS: Maternal weight gain, body mass index at term, smoking and hemoglobin values were significantly higher in breech presentation than in cephalic pregnancies. The placenta was located in the cornu-fundal region in 63.2% of breech presentations and 26.8% of cephalic presentations (p<0.001). Placental weights were 657 g and 597 g, respectively (p<0.001). Umbilical cord length was shorter in breech than cephalic pregnancies (p<0.001). Although breech pregnancies had significantly reduced fetal heart rate variability (p<0.001), Apgar scores were much higher in breech fetuses than in cephalic fetuses. Ninety-five percent of breech pregnancies underwent cesarean sections. CONCLUSION: Cornu-fundal localization of the placenta, smoking, greater maternal weight gain, higher body mass index at term, greater placental weight, shorter umbilical cord, and lower estimated fetal weight may be predictive of persistent breech presentation. Reduced fetal heart rate variability did not have an adverse effect on Apgar scores after cesarean delivery in breech fetuses with no other problems at term. PMID- 19126506 TI - High-dose misoprostol as an alternative therapy after failed medical abortion. AB - OBJECTIVE: The aim of this study was to determine the complete abortion rate for the vaginal administration of high-dose misoprostol after a failed medical abortion. MATERIALS AND METHODS: When their medical abortions failed after the conventional oral administration of mifepristone and misoprostol, participants then received 1,000 microg of misoprostol vaginally. The efficacy and side effects of this treatment were evaluated. RESULTS: Twenty-seven women who failed to abort after the conventional administration of mifepristone and misoprostol were enrolled in this trial. Fourteen days after the vaginal administration of 1,000 microg misoprostol, the overall complete expulsion rate had reached 88.8% (24/27). Most adverse effects were mild to moderate and did not require treatment. CONCLUSION: The vaginal administration of 1,000 microg misoprostol as a salvage therapy after a failed medical abortion appears to be a safe and highly effective alternative to surgical intervention. PMID- 19126507 TI - Diagnosis of human papillomavirus infection by abnormal cervical cytology is highly reproducible after vaginal douching. AB - OBJECTIVE: To evaluate whether human papillomavirus (HPV) activity in symptomatic patients with abnormal Papanicolaou smears (minimal abnormal cytology) was affected by a complete vaginal douche, applied to the cervix prior to colposcopic examination. MATERIALS AND METHODS: A total of 132 women with abnormal Papanicolaou smears were prospectively registered in this study from 1999 to 2004. Two specimens were collected from each patient; one before and one after the cervix was swabbed with a complete vaginal douche during a colposcopic examination. We compared the HPV detection results before and after douching in the same patients. The samples were analyzed using the Hybrid Capture II assay. RESULTS: Before douching, 57.6% (76/132) of the samples were HPV-positive, compared with 58.3% (77/132) of the samples after douching. There was a strong correlation between the cervical HPV detection rates between the before and after douche samples (p<0.001). The kappa statistic was 0.891 for the correlation (positive- positive, negative-negative) and the discrepancy (positive-negative) between the inter-douche rates of HPV activity. A comparison of the HPV assay before and after vaginal douching demonstrated a statistically significant relationship (p<0.001). The sensitivity of the HPV test after douching was higher (97.8%) than that before douching (95.6%). The false-negative rate before douching in women with inflammatory smears was 3% (4/132) and the false-positive rate before douching in women with atypical squamous cells of undetermined significance was 2.1% (3/132). CONCLUSION: Our results indicate that self administration of a vaginal douche is not a reliable method of eradicating HPV infection. PMID- 19126508 TI - Factors affecting voiding function in urogynecology patients. AB - OBJECTIVE: Our objective was to assess the impacts of menopause, age and parity on voiding function in urogynecology patients. MATERIALS AND METHODS: The medical records of 4,470 patients with urodynamic studies were reviewed at our center between January 1999 and May 2003. Patients with conditions including a prior continence procedure, advanced pelvic prolapse, hysterectomy or neurologic deficits were excluded from our study. Comprehensive medical histories, physical examinations, bladder diaries and results of multi-channel urodynamic testing were analyzed. RESULTS: The mean age of the 3,161 women enrolled was 50 years, and the mean parity was three. Four urodynamic variables served as voiding parameters: maximum flow rate (Qmax) and post-void residual (PVR) from uroflowmetry, and maximum flow rate (Qmax.p) and detrusor pressure at maximum flow (PdetQmax) from pressure-flow studies. Of the three factors studied (menopause, age, and parity), only parity had a significant impact on uroflowmetry and pressure-flow study results (Qmax, p=0.007; PVR, p<0.001; Qmax.p, p=0.002; PdetQmax, p<0.001). Twenty-five percent of the patients in our study were diagnosed with voiding dysfunction. CONCLUSION: Our results indicated that parity had a significant impact on voiding function in urogynecology patients. Twenty-five percent of patients in our study were diagnosed with voiding dysfunction. The bladder behavior in women after childbirth may be more complex than previously thought, and special attention should be paid to women who suffer from bladder symptoms after childbirth. PMID- 19126510 TI - Successful conservative management of placenta previa totalis and extensive percreta. PMID- 19126509 TI - Genetic network analysis of human CD34+ hematopoietic stem/precursor cells. AB - OBJECTIVE: Somatic CD34+ hematopoietic stem/precursor cells (HSPCs) give rise to hematopoietic cells and endothelial cells and have been used in clinical applications. Understanding the genes responsible for stemness and how they interact with each other will help us to manipulate these cells more efficiently in the future. MATERIALS AND METHODS: We performed microarray analysis on human CD34+ HSPCs and on two different progeny cell types, i.e. microvascular endothelial cells and peripheral blood mononuclear cells. Systems biology and advanced bioinformatics tools were used to help clarify the genetic networks associated with these stem cell genes. RESULTS: We identified CD34+ HSPC genes and found that they were involved in critical biologic processes such as cell cycle regulation, chromosome organization, and DNA repair. We also identified a novel precursor gene cluster on chromosome 19p13.3. Analysis of HSPC-enriched genes using systems biology tools revealed a complex genetic network functioning in CD34+ cells, in which several genes acted as hubs to maintain the stability (such as GATA1) or connectivity (such as hepatic growth factor) of the whole network. CONCLUSION: This study provides the foundation for a more detailed understanding of CD34+ HSPCs. PMID- 19126511 TI - Neonatal coagulopathy presents as unusual and severe subgaleal hematomas after vacuum delivery. PMID- 19126512 TI - Early prenatal diagnosis of semilobar holoprosencephaly combined with a dorsal cyst and no facial defect. PMID- 19126513 TI - Prenatal diagnosis of DiGeorge syndrome. PMID- 19126514 TI - Cervical pregnancy treated with systemic methotrexate administration and resectoscopy. PMID- 19126515 TI - Uterine perforation and bowel incarceration following surgical abortion during the first trimester. PMID- 19126516 TI - Conservative laparoscopy following prophylactic methotrexate for an unruptured bilateral tubal pregnancy. PMID- 19126517 TI - Recurrent uterine leiomyosarcoma implanted in a laparotomy scar. PMID- 19126518 TI - Primary vaginal stone in a young active woman. PMID- 19126519 TI - Intracerebral hemorrhage with cerebral arteriovenous malformation rupture during pregnancy. PMID- 19126520 TI - Outcome of inadvertently performed hysterosalpingography during early pregnancy-- 7 years after birth. PMID- 19126521 TI - The use of a concomitant tension-free vaginal mesh technique and a tension-free midurethral sling in treating pelvic organ prolapse and occult stress urinary incontinence. PMID- 19126522 TI - Is ovarian pregnancy a medical illness? Methotrexate treatment failure and rescue by laparoscopic removal. PMID- 19126523 TI - Limb-body wall complex with craniofacial defects after ovarian stimulation. PMID- 19126524 TI - Concomitant exencephaly and limb defects associated with pentalogy of Cantrell. PMID- 19126525 TI - First- and Second-trimester Down Syndrome Screening: Current Strategies and Clinical Guidelines. PMID- 19126527 TI - Feeding muscles scale differently from swimming muscles in sunfish (Centrarchidae). AB - The physiological properties of vertebrate skeletal muscle typically show a scaling pattern of slower contractile properties with size. In fishes, the myotomal or swimming muscle reportedly follows this pattern, showing slower muscle activation, relaxation and maximum shortening velocity (V(max)) with an increase in body size. We asked if the muscles involved in suction feeding by fishes would follow the same pattern. We hypothesized that feeding muscles in fishes that feed on evasive prey are under selection to maintain high power output and therefore would not show slower contractile properties with size. To test this, we compared contractile properties in feeding muscles (epaxial and sternohyoideus) and swimming muscle (myotomal) for two members of the family Centrarchidae (sunfish): the bluegill (Lepomis macrochirus) and the largemouth bass (Micropterus salmoides). Consistent with our predictions, the V(max) of myotomal muscle in both species slowed with size, while the epaxials showed no significant change in V(max) with size. In the sternohyoideus, V(max) slowed with size in the bluegill but increased with size in the bass. The results indicate that scaling patterns of contractile properties appear to be more closely tied to muscle function (i.e. locomotion versus feeding) than overall patterns of size. PMID- 19126528 TI - Marine hybrid hotspot at Indo-Pacific biogeographic border. AB - Studying hybridization is crucial to understanding speciation and almost all our knowledge comes from terrestrial and freshwater environments. Marine hybrids are considered rare, particularly on species-rich coral reefs. Here, we report a significant marine hybrid zone at Christmas and Cocos Islands (eastern Indian Ocean) with 11 hybrid coral reef fishes (across six families); the most recorded hybrids of any marine location. In most cases, at least one of the parent species is rare (less than three individuals per 3000 m(2)), suggesting that hybridization has occurred because individuals of the rare species have mated with another species owing to a scarcity of conspecific partners. These islands also represent a marine suture zone where many of the hybrids have arisen through interbreeding between Indian and Pacific Ocean species. For these species, it appears that past climate changes allowed species to diverge in allopatry, while recent conditions have facilitated contact and subsequent hybridization at this Indo-Pacific biogeographic border. The discovery of the Christmas-Cocos hybrid zone refutes the notion that hybridization is lacking on coral reefs and provides a natural laboratory for testing the generality of terrestrially derived hybridization theory in the marine environment. PMID- 19126529 TI - Calculated reciprocity after all: computation behind token transfers in orang utans. AB - Transfers and services are frequent in the animal kingdom. However, there is no clear evidence in animals that such transactions are based on weighing costs and benefits when giving or returning favours and keeping track of them over time (i.e. calculated reciprocity). We tested two orang-utans (Pongo pygmaeus abelii) in a token-exchange paradigm, in which each individual could exchange a token for food with the experimenter but only after first obtaining the token from the other orang-utan. Each orang-utan possessed tokens valuable to their partner but useless to themselves. Both orang-utans actively transferred numerous tokens (mostly partner-valuable) to their partner. One of the orang-utans routinely used gestures to request tokens while the other complied with such requests. Although initially the transfers were biased in one direction, they became more balanced towards the end of the study. Indeed, data on the last three series produced evidence of reciprocity both between and within trials. We observed an increase in the number and complexity of exchanges and alternations. This study is the first experimental demonstration of the occurrence of direct transfers of goods based on calculated reciprocity in non-human-primates. PMID- 19126530 TI - Evidence for egg discrimination preceding failed rejection attempts in a small cuckoo host. AB - Given the high costs of avian obligate brood parasitism, host individuals are selected to reject parasitic eggs they recognize as foreign. We show that rejection may not necessarily follow egg discrimination when selective removal of the parasitic egg is difficult. We studied egg rejection behaviour in a small host of the common cuckoo Cuculus canorus, the eastern olivaceous warbler Hippolais pallida, by experimental parasitism with model and real non-mimetic cuckoo eggs and video recordings of host behaviour. Hosts pecked 87 per cent (20 out of 23) of the model eggs but eventually accepted 43.5 per cent (10 out of 23) of them. A similar pattern was found for real cuckoo eggs, which were all pecked, but as many as 47 per cent (7 out of 15) of them were accepted. To our knowledge, this is the first demonstration of a cuckoo host discriminating against real parasitic eggs but often accepting them. Our results also show that in host species experiencing difficulties in performing puncture ejection, non-mimetic cuckoo eggs may avoid rejection by means of their unusually high structural strength. PMID- 19126531 TI - A rebuttal to the claim natural beaches confer fitness benefits to nesting marine turtles. PMID- 19126532 TI - Parental effects on offspring life histories: when are they important? PMID- 19126533 TI - Indirect cues of nest predation risk and avian reproductive decisions. AB - Current life-history theory predicts that increased mortality at early stages of life leads to reduced initial investment (e.g. clutch size) but increased subsequent investment during the reproduction attempt. In a field experiment, migratory pied flycatchers Ficedula hypoleuca perceived differences in mammalian nest predation risk and altered their reproductive strategies in two respects. First, birds avoided nest sites manipulated to reflect the presence of a predator. Second, birds breeding in risky areas nested 4 days earlier and laid 10 per cent larger clutches than those in safe areas, a result that runs counter to the prevailing life-history paradigm. We suggest that the overwhelming importance of nest predation to individual fitness reduces the value of collecting other information on habitat features leading to expedited onset of nesting, and, consequently, to larger clutch size. PMID- 19126534 TI - Marine mammals' influence on ecosystem processes affecting fisheries in the Barents Sea is trivial. AB - Some interpretations of ecosystem-based fishery management include culling marine mammals as an integral component. The current Norwegian policy on marine mammal management is one example. Scientific support for this policy includes the Scenario Barents Sea (SBS) models. These modelled interactions between cod, Gadus morhua, herring, Clupea harengus, capelin, Mallotus villosus and northern minke whales, Balaenoptera acutorostrata. Adding harp seals Phoca groenlandica into this top-down modelling approach resulted in unrealistic model outputs. Another set of models of the Barents Sea fish-fisheries system focused on interactions within and between the three fish populations, fisheries and climate. These model key processes of the system successfully. Continuing calls to support the SBS models despite their failure suggest a belief that marine mammal predation must be a problem for fisheries. The best available scientific evidence provides no justification for marine mammal culls as a primary component of an ecosystem based approach to managing the fisheries of the Barents Sea. PMID- 19126535 TI - The movement of motion-defined contours can bias perceived position. AB - Illusory position shifts induced by motion suggest that motion processing can interfere with perceived position. This may be because accurate position representation is lost during successive visual processing steps. We found that complex motion patterns, which can only be extracted at a global level by pooling and segmenting local motion signals and integrating over time, can influence perceived position. We used motion-defined Gabor patterns containing motion defined boundaries, which themselves moved over time. This 'motion-defined motion' induced position biases of up to 0.5 degrees , much larger than has been found with luminance-defined motion. The size of the shift correlated with how detectable the motion-defined motion direction was, suggesting that the amount of bias increased with the magnitude of this complex directional signal. However, positional shifts did occur even when participants were not aware of the direction of the motion-defined motion. The size of the perceptual position shift was greatly reduced when the position judgement was made relative to the location of a static luminance-defined square, but not eliminated. These results suggest that motion-induced position shifts are a result of general mechanisms matching dynamic object properties with spatial location. PMID- 19126536 TI - Modelling control of epidemics spreading by long-range interactions. AB - We have studied the spread of epidemics characterized by a mixture of local and non-local interactions. The infection spreads on a two-dimensional lattice with the fixed nearest neighbour connections. In addition, long-range dynamical links are formed by moving agents (vectors). Vectors perform random walks, with step length distributed according to a thick-tail distribution. Two distributions are considered in this paper, an alpha-stable distribution describing self-similar vector movement, yet characterized by an infinite variance and an exponential power characterized by a large but finite variance. Such long-range interactions are hard to track and make control of epidemics very difficult. We also allowed for cryptic infection, whereby an infected individual on the lattice can be infectious prior to showing any symptoms of infection or disease. To account for such cryptic spread, we considered a control strategy in which not only detected, i.e. symptomatic, individuals but also all individuals within a certain control neighbourhood are treated upon the detection of disease. We show that it is possible to eradicate the disease by using such purely local control measures, even in the presence of long-range jumps. In particular, we show that the success of local control and the choice of the optimal strategy depend in a non-trivial way on the dispersal patterns of the vectors. By characterizing these patterns using the stability index of the alpha-stable distribution to change the power law behaviour or the exponent characterizing the decay of an exponential power distribution, we show that infection can be successfully contained using relatively small control neighbourhoods for two limiting cases for long-distance dispersal and for vectors that are much more limited in their dispersal range. PMID- 19126537 TI - p38 MAPK and beta-arrestin 2 mediate functional interactions between endogenous micro-opioid and alpha2A-adrenergic receptors in neurons. AB - Formation of receptor complexes between micro-opioid and alpha2A-adrenergic receptors has been demonstrated in transfected cells. The functional significance and underlying mechanisms of such receptor interactions remain to be determined in neuronal systems. We examined functional interactions between endogenous micro and alpha2A receptors in mouse dorsal root ganglion neurons. Acute application of the micro agonist [D-Ala2,N-MePhe4, Gly-ol5]enkephalin (DAMGO) or the alpha2 agonist clonidine inhibited voltage-gated Ca2+ currents in these neurons. Prolonged treatment with either DAMGO or clonidine induced a mutual cross desensitization between micro and alpha2A receptor-mediated current inhibition. The cross-desensitization was closely associated with simultaneous internalization of micro and alpha2A receptors. Morphine, a mu agonist triggering little mu receptor endocytosis, induced neither cross-desensitization nor internalization of alpha2A receptors. Furthermore, inhibition of p38 MAPK prevented the cross-desensitization as well as cointernalization of micro and alpha2A receptors. Changes in receptor trafficking profiles suggested that p38 MAPK activity was required for initiating micro receptor internalization and maintaining possible micro-alpha2A association during their cointernalization. Finally, the micro-alpha2A cross-desensitization was absent in dorsal root ganglion neurons lacking beta-arrestin 2. These findings demonstrated p38 MAPK- and beta-arrestin 2-dependent cross-regulation between neuronal micro and alpha2A receptors. By promoting receptor cross-desensitization and cointernalization, such functional interactions may serve as negative feedback mechanisms triggered by prolonged agonist exposure to modulate the signaling of functionally related G protein-coupled receptors. PMID- 19126538 TI - Mobilization of lysosomal calcium regulates the externalization of phosphatidylserine during apoptosis. AB - A hallmark of apoptotic cells is the Ca2+-dependent appearance of phosphatidylserine (PS) at the cell surface as a result of its redistribution from the inner-to-outer plasma membrane leaflet. Although endoplasmic reticulum and mitochondrial Ca2+ are known to participate in apoptosis, their role in PS externalization has not been established. In this study, several organelle specific fluorescent markers and Ca2+-sensitive probes were used to identify the source of Ca2+ critical to PS externalization. By employing Rhod-2AM, fluorescein labeled high molecular weight dextran, and Calcium Green 1, we provide evidence that lysosomes respond to apoptotic stimuli by releasing their luminal Ca2+ to the cytosol. Cells treated with the cytosolic phospholipase A2 inhibitor, cPLA2alpha, had no effect on caspase activation but exhibited a significant decrease in lysosomal Ca2+ release and externalization of PS in response to apoptotic stimuli. Similarly, cells depleted of lysosomal Ca2+ underwent programmed cell death yet failed to externalize PS. These data indicate that although Ca2+ release from other intracellular organelles to the cytosol is adequate for apoptosis, the release of Ca2+ from lysosomes is critical for PS externalization. PMID- 19126539 TI - Identification of plasmin-interactive sites in the light chain of factor VIII responsible for proteolytic cleavage at Lys36. AB - We have recently reported that plasmin likely associates with the factor VIII light chain to proteolyze at Lys36 within the A1 domain. In this study, we determined that the rate of plasmin-catalyzed inactivation on the forms of factor VIIIa containing A1-(1-336) and 1722A3C1C2, reflecting Lys36 cleavage, was reduced by approximately 60%, compared with those containing 1649A3C1C2 and 1690A3C1C2. SDS-PAGE analysis revealed that Lys36 cleavage of factor VIIIa with 1722A3C1C2 was markedly slower than those with 1649A3C1C2 and 1690A3C1C2. Surface plasmon resonance-based assays, using active site-modified anhydro-plasmin (Ah plasmin) showed that 1722A3C1C2 bound to Ah-plasmin with an approximately 3-fold lower affinity than 1649A3C1C2 or 1690A3C1C2 (Kd, 176, 68.2, and 60.3 nM, respectively). Recombinant A3 bound to Ah-plasmin (Kd, 44.2 nM), whereas C2 failed to bind, confirming the presence of a plasmin-binding site within N terminus of A3. Furthermore, the Glu-Gly-Arg active site-modified factor IXa also blocked 1722A3C1C2 binding to Ah-plasmin by approximately 95%, supporting the presence of another plasmin-binding site overlapping the factor IXa-binding site in A3. In keeping with a major contribution of the lysine-binding sites in plasmin for interaction with the factor VIII light chain, analysis of the A3 sequence revealed two regions involving clustered lysine residues in 1690-1705 and 1804-1818. Two peptides based on these regions blocked 1649A3C1C2 binding to Ah-plasmin by approximately 60% and plasmin-catalyzed Lys36 cleavage of factor VIIIa with A1-(1-336) by approximately 80%. Our findings indicate that an extended surface, centered on residues 1690-1705 and 1804-1818 within the A3 domain, contributes to a unique plasmin-interactive site that promotes plasmin docking during cofactor inactivation by cleavage at Lys36. PMID- 19126540 TI - Calmodulin-driven nuclear entry: trigger for sex determination and terminal differentiation. AB - We originally proposed that Ca(2+)-calmodulin mediates a novel nuclear entry pathway distinct from the canonic Ran-dependent pathway (Sweitzer, T. D., and Hanover, J. A. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 14574-14579). Although seemingly redundant, Ca(2+)-calmodulin-driven nuclear entry is now known to facilitate nuclear delivery of architectural transcription factors to chromatin. Intriguingly, defects in calmodulin-driven nuclear import of the transcription factors SRY and SOX9 in Sertoli cells lead to human sex reversal diseases with altered male gonad development. Calmodulin-triggered nuclear entry is an evolutionarily ancient feature of eukaryotes observed from yeast to man. Ca(2+) calmodulin-triggered nuclear entry of key architectural transcription factors is a potentially key epigenetic regulator of terminal differentiation in response to cell signaling. PMID- 19126541 TI - Deleted in breast cancer 1, a novel androgen receptor (AR) coactivator that promotes AR DNA-binding activity. AB - Androgen receptor (AR) plays a critical role in development and maintenance of male reproductive functions and the etiology of prostate cancer. As a ligand regulated transcription factor, identification and characterization of AR coregulators are essential for understanding the molecular mechanisms underlying its diverse biological functions. Here we reported the identification of a novel AR coactivator, deleted in breast cancer 1 (DBC1), through a biochemical approach. DBC1 interacts with AR in a ligand-stimulated manner and facilitates AR transcriptional activation in transfected cells as well as in Xenopus oocytes. In in vitro gel shift experiments, recombinant DBC1 drastically enhanced AR DNA binding activity. Expression of DBC1 also enhanced the binding of AR to chromatinized template in vivo, whereas knockdown of DBC1 impaired the binding of AR to endogenous prostate-specific antigen (PSA) gene in the prostate cancer cell line LNCaP. Thus, our data identify DBC1 as a novel AR coactivator. PMID- 19126542 TI - Curvature dynamics of alpha-synuclein familial Parkinson disease mutants: molecular simulations of the micelle- and bilayer-bound forms. AB - Alpha-synuclein remains a protein of interest due to its propensity to form fibrillar aggregates in neurodegenerative disease and its putative function in synaptic vesicle regulation. Herein, we present a series of atomistic molecular dynamics simulations of wild-type alpha-synuclein and three Parkinson disease familial mutants (A30P, A53T, and E46K) in two distinct environments. First, in order to match recent NMR experiments, we have simulated each protein bound to an SDS detergent micelle. Second, in order to connect more closely to the true biological environment, we have simulated the proteins bound to a 1,2-dioleoyl-sn glycero-3-phosphoserine lipid bilayer. In the micelle-bound case, we find that the wild type and all of the variants of alpha-synuclein flatten the underlying micelle, decreasing its surface area. A30P is known to lessen alpha synuclein/membrane affinity and, consistent with experiment, destabilizes the simulated secondary structure. In the case of A53T, our simulations reveal a range of stabilizing hydrogen bonds that form with the threonine. In both environments, the E46K mutation, which is known to increase bilayer affinity, leads to an additional hydrogen bond between the protein and either the detergent or lipid. Simulations indicate that alphaS and its variants are less dynamic in the bilayer than in the micelle. Furthermore, the simulations of the mutants suggest how changes in the structure and dynamics of alpha-synuclein may affect its biological role. PMID- 19126544 TI - A phosphorylation cascade controls the degradation of active SREBP1. AB - Sterol regulatory element-binding proteins (SREBPs) are a family of transcription factors that regulates cholesterol and lipid metabolism. The active forms of these transcription factors are targeted by a number of post-translational modifications, including phosphorylation. Phosphorylation of Thr-426 and Ser-430 in SREBP1a creates a docking site for the ubiquitin ligase Fbw7, resulting in the degradation of the transcription factor. Here, we identify a novel phosphorylation site in SREBP1a, Ser-434, which regulates the Fbw7-dependent degradation of SREBP1. We demonstrate that both SREBP1a and SREBP1c are phosphorylated on this residue (Ser-410 in SREBP1c). Importantly, we demonstrate that the mature form of endogenous SREBP1 is phosphorylated on Ser-434. Glycogen synthase kinase-3 phosphorylates Ser-434, and the phosphorylation of this residue is attenuated in response to insulin signaling. Interestingly, phosphorylation of Ser-434 promotes the glycogen synthase kinase-3-dependent phosphorylation of Thr 426 and Ser-430 and destabilizes SREBP1. Consequently, mutation of Ser-434 blocks the interaction between SREBP1 and Fbw7 and attenuates Fbw7-dependent degradation of SREBP1. Importantly, insulin fails to enhance the levels of mature SREBP1 in cells lacking Fbw7. Thus, the degradation of mature SREBP1 is controlled by cross talk between multiple phosphorylated residues in its C-terminal domain and the phosphorylation of Ser-434 could function as a molecular switch to control these processes. PMID- 19126543 TI - Adipocyte-specific expression of murine resistin is mediated by synergism between peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding proteins. AB - Resistin antagonizes insulin action in mouse, making it a potential therapeutic target for treating metabolic diseases such as diabetes. To better understand how mouse resistin gene (Retn) expression is restricted to fat tissue, we identified an adipocyte-specific enhancer located approximately 8.8-kb upstream of the transcription start site. This region contains a binding site for the master adipogenic regulator peroxisome proliferator-activated receptor gamma (PPARgamma), and binds endogenous PPARgamma together with its partner retinoid-X receptor alpha (RXRalpha). It also contains three binding sites for CCAAT/enhancer-binding protein (C/EBP), and is bound by endogenous C/EBPalpha and C/EBPbeta in adipocytes. Exogenous expression of PPARgamma/RXRalpha and C/EBPalpha in non-adipocyte cells synergistically drives robust expression from the enhancer. Although PPARgamma ligands repress Retn transcription in adipocytes, rosiglitazone paradoxically stimulates the enhancer activity, suggesting that the enhancer is not directly involved in negative regulation. Unlike expression of Retn in mouse, human resistin (RETN) is expressed primarily in macrophages. Interestingly, the region homologous to the mouse Retn enhancer in the human gene contains all three C/EBP elements, but is not conserved for the sequence bound by PPARgamma. Furthermore, it displays little or no binding by PPARgamma in vitro. Taken together, the data suggest that a composite enhancer binding both PPARgamma and C/EBP factors confers adipocyte-specific expression to Retn in mouse, and its absence from the human gene may explain the lack of adipocyte expression in humans. PMID- 19126545 TI - The green-absorbing Drosophila Rh6 visual pigment contains a blue-shifting amino acid substitution that is conserved in vertebrates. AB - The molecular mechanisms that regulate invertebrate visual pigment absorption are poorly understood. Through sequence analysis and functional investigation of vertebrate visual pigments, numerous amino acid substitutions important for this adaptive process have been identified. Here we describe a serine/alanine (S/A) substitution in long wavelength-absorbing Drosophila visual pigments that occurs at a site corresponding to Ala-292 in bovine rhodopsin. This S/A substitution accounts for a 10-17-nm absorption shift in visual pigments of this class. Additionally, we demonstrate that substitution of a cysteine at the same site, as occurs in the blue-absorbing Rh5 pigment, accounts for a 4-nm shift. Substitutions at this site are the first spectrally significant amino acid changes to be identified for invertebrate pigments sensitive to visible light and are the first evidence of a conserved tuning mechanism in vertebrate and invertebrate pigments of this class. PMID- 19126546 TI - Structural and functional analysis of the C-terminal DNA binding domain of the Salmonella typhimurium SPI-2 response regulator SsrB. AB - In bacterial pathogenesis, virulence gene regulation is controlled by two component regulatory systems. In Escherichia coli, the EnvZ/OmpR two-component system is best understood as regulating expression of outer membrane proteins, but in Salmonella enterica, OmpR activates transcription of the SsrA/B two component system located on Salmonella pathogenicity island 2 (SPI-2). The response regulator SsrB controls expression of a type III secretory system in which effectors modify the vacuolar membrane and prevent its degradation via the endocytic pathway. Vacuolar modification enables Salmonella to survive and replicate in the macrophage phagosome and disseminate to the liver and spleen to cause systemic infection. The signals that activate EnvZ and SsrA are unknown but are related to the acidic pH encountered in the vacuole. Our previous work established that SsrB binds to regions of DNA that are AT-rich, with poor sequence conservation. Although SsrB is a major virulence regulator in Salmonella, very little is known regarding how it binds DNA and activates transcription. In the present work, we solved the structure of the C-terminal DNA binding domain of SsrB (SsrB(C)) by NMR and analyzed the effect of amino acid substitutions on function. We identified residues in the DNA recognition helix (Lys(179), Met(186)) and the dimerization interface (Val(197), Leu(201)) that are important for SsrB transcriptional activation and DNA binding. An essential cysteine residue in the N-terminal receiver domain was also identified (Cys(45)), and the effect of Cys(203) on dimerization was evaluated. Our results suggest that although disulfide bond formation is not required for dimerization, dimerization occurs upon DNA binding and is required for subsequent activation of transcription. Disruption of the dimer interface by a C203E substitution reduces SsrB activity. Modification of Cys(203) or Cys(45) may be an important mode of SsrB inactivation inside the host. PMID- 19126547 TI - Functional characterization and substrate specificity of spinosyn rhamnosyltransferase by in vitro reconstitution of spinosyn biosynthetic enzymes. AB - Spinosyn, a potent insecticide, is a novel tetracyclic polyketide decorated with d-forosamine and tri-O-methyl-L-rhamnose. Spinosyn rhamnosyltransferase (SpnG) is a key biocatalyst with unique sequence identity and controls the biosynthetic maturation of spinosyn. The rhamnose is critical for the spinosyn insecticidal activity and cell wall biosynthesis of the spinosyn producer, Saccharopolyspora spinosa. In this study, we have functionally expressed and characterized SpnG and the three enzymes, Gdh, Epi, and Kre, responsible for dTDP-L-rhamnose biosynthesis in S. spinosa by purified enzymes from Escherichia coli. Most notably, the substrate specificity of SpnG was thoroughly characterized by kinetic and inhibition experiments using various NDP sugar analogs made by an in situ combination of NDP-sugar-modifying enzymes. SpnG was found to exhibit striking substrate promiscuity, yielding corresponding glycosylated variants. Moreover, the critical residues presumably involved in catalytic mechanism of Gdh and SpnG were functionally evaluated by site-directed mutagenesis. The information gained from this study has provided important insight into molecular recognition and mechanism of the enzymes, especially SpnG. The results have made possible the structure-activity characterization of SpnG, as well as the use of SpnG or its engineered form to serve as a combinatorial tool to make spinosyn analogs with altered biological activities and potency. PMID- 19126548 TI - Growth factors outside of the platelet-derived growth factor (PDGF) family employ reactive oxygen species/Src family kinases to activate PDGF receptor alpha and thereby promote proliferation and survival of cells. AB - The vitreous contains a plethora of growth factors that are strongly implicated in the formation of fibroproliferative diseases such as proliferative vitreoretinopathy. Although platelet-derived growth factors (PDGFs) are present in the vitreous, vitreal growth factors outside of the PDGF family activated the PDGF alpha receptor (PDGFRalpha) and promoted disease progression in a rabbit model of proliferative vitreoretinopathy (H. Lei, G. Velez, P. Hovland, T. Hirose, D. Gilbertson, and A. Kazlauskas (2008) submitted for publication.) In this report we investigated the mechanism by which non-PDGFs activated PDGFRalpha. We found that non-PDGFs increased the cellular level of reactive oxygen species (ROS) and that this event was necessary and sufficient for phosphorylation of PDGFRalpha. We speculated that the underlying mechanism was ROS-mediated inhibition of phosphotyrosine phosphatases, which antagonize receptor auto-phosphorylation. However, this did not appear to be the case. Non PDGFs promoted tyrosine phosphorylation of catalytically inactive PDGFRalpha, and thereby indicated that at least one additional tyrosine kinase was involved. Indeed, preventing expression or blocking the kinase activity of Src family kinases suppressed non-PDGF-dependent tyrosine phosphorylation of PDGFRalpha. Thus non-PDGFs increased the level of ROS, which activated Src family kinases and resulted in phosphorylation of PDGFRalpha. Finally, although non-PDGFs induced only modest phosphorylation of PDGFRalpha, proliferation and survival of cells in response to non-PDGFs was significantly enhanced by expression of PDGFRalpha. These studies reveal a novel mechanism for activation of PDGFRalpha that appears capable of enhancing the responsiveness of cells to growth factors outside of the PDGF family. PMID- 19126550 TI - let-7 Overexpression leads to an increased fraction of cells in G2/M, direct down regulation of Cdc34, and stabilization of Wee1 kinase in primary fibroblasts. AB - microRNAs play a critically important role in a wide array of biological processes including those implicated in cancer, neuro-degenerative and metabolic disorders, and viral infection. Although we have begun to understand microRNA biogenesis and function, experimental demonstration of their functional effects and the molecular mechanisms by which they function remains a challenge. Members of the let-7/miR-98 family play a critical role in cell cycle control with respect to differentiation and tumorigenesis. In this study, we show that exogenous addition of pre-let-7 in primary human fibroblasts results in a decrease in cell number and an increased fraction of cells in the G(2)/M cell cycle phase. Combining microarray techniques with DNA sequence analysis to identify potential let-7 targets, we discovered 838 genes with a let-7 binding site in their 3'-untranslated region that were down-regulated upon overexpression of let-7b. Among these genes is cdc34, the ubiquitin-conjugating enzyme of the Skp1/cullin/F-box (SCF) complex. Cdc34 protein levels are strongly down-regulated by let-7 overexpression. Reporter assays demonstrated direct regulation of the cdc34 3'-untranslated region by let-7. We hypothesized that low Cdc34 levels would result in decreased SCF activity, stabilization of the SCF target Wee1, and G(2)/M accumulation. Consistent with this hypothesis, small interfering RNA mediated down-regulation of Wee1 reversed the G(2)/M phenotype induced by let-7 overexpression. We conclude that Cdc34 is a functional target of let-7 and that let-7 induces down-regulation of Cdc34, stabilization of the Wee1 kinase, and an increased fraction of cells in G(2)/M in primary fibroblasts. PMID- 19126551 TI - User evaluation of a communication system that automatically generates captions to improve telephone communication. AB - This study examined the subjective benefit obtained from automatically generated captions during telephone-speech comprehension in the presence of babble noise. Short stories were presented by telephone either with or without captions that were generated offline by an automatic speech recognition (ASR) system. To simulate online ASR, the word accuracy (WA) level of the captions was 60% or 70% and the text was presented delayed to the speech. After each test, the hearing impaired participants (n = 20) completed the NASA-Task Load Index and several rating scales evaluating the support from the captions. Participants indicated that using the erroneous text in speech comprehension was difficult and the reported task load did not differ between the audio + text and audio-only conditions. In a follow-up experiment (n = 10), the perceived benefit of presenting captions increased with an increase of WA levels to 80% and 90%, and elimination of the text delay. However, in general, the task load did not decrease when captions were presented. These results suggest that the extra effort required to process the text could have been compensated for by less effort required to comprehend the speech. Future research should aim at reducing the complexity of the task to increase the willingness of hearing impaired persons to use an assistive communication system automatically providing captions. The current results underline the need for obtaining both objective and subjective measures of benefit when evaluating assistive communication systems. PMID- 19126549 TI - Regulation of epithelial Na+ transport by soluble adenylyl cyclase in kidney collecting duct cells. AB - Alkalosis impairs the natriuretic response to diuretics, but the underlying mechanisms are unclear. The soluble adenylyl cyclase (sAC) is a chemosensor that mediates bicarbonate-dependent elevation of cAMP in intracellular microdomains. We hypothesized that sAC may be an important regulator of Na(+) transport in the kidney. Confocal images of rat kidney revealed specific immunolocalization of sAC in collecting duct cells, and immunoblots confirmed sAC expression in mouse cortical collecting duct (mpkCCD(c14)) cells. These cells exhibit aldosterone stimulated transepithelial Na(+) currents that depend on both the apical epithelial Na(+) channel (ENaC) and basolateral Na(+),K(+)-ATPase. RNA interference-mediated 60-70% knockdown of sAC expression comparably inhibited basal transepithelial short circuit currents (I(sc)) in mpkCCD(c14) cells. Moreover, the sAC inhibitors KH7 and 2-hydroxyestradiol reduced I(sc) in these cells by 50-60% within 30 min. 8-Bromoadenosine-3',5'-cyclic-monophosphate substantially rescued the KH7 inhibition of transepithelial Na(+) current. Aldosterone doubled ENaC-dependent I(sc) over 4 h, an effect that was abolished in the presence of KH7. The sAC contribution to I(sc) was unaffected with apical membrane nystatin-mediated permeabilization, whereas the sAC-dependent Na(+) current was fully inhibited by basolateral ouabain treatment, suggesting that the Na(+),K(+)-ATPase, rather than ENaC, is the relevant transporter target of sAC. Indeed, neither overexpression of sAC nor treatment with KH7 modulated ENaC currents in Xenopus oocytes. ATPase and biotinylation assays in mpkCCD(c14) cells demonstrated that sAC inhibition decreases catalytic activity rather than surface expression of the Na(+),K(+)-ATPase. In summary, these results suggest that sAC regulates both basal and agonist-stimulated Na(+) reabsorption in the kidney collecting duct, acting to enhance Na(+),K(+)-ATPase activity. PMID- 19126552 TI - The bilateral pulmonary artery banding for hypoplastic left heart syndrome with a diminutive ascending aorta. AB - A one-day-old neonate who was diagnosed with hypoplastic left heart syndrome (HLHS), aortic atresia, with a diminutive ascending aorta, and mitral atresia, was referred to us for cardiogenic shock because of excessive pulmonary blood flow. The patient underwent bilateral pulmonary artery banding (bPAB). After bPAB, the patient's hemodynamics were still unstable because of coronary malperfusion, to proceed to undergo Norwood procedure at the age of 3 days. In this case, the stenosis of the ascending aorta, just proximal to the innominate artery caused coronary ischemia. The precise evaluation of the ascending aorta is necessary to perform the bPAB for HLHS with diminutive ascending aorta. If there is a sign of stenosis of the ascending aorta, the Norwood procedure should be performed as the first stage palliation, even for high-risk HLHS patients. PMID- 19126553 TI - Extra-anatomic bypass for recurrent abdominal aortic and renal in-stent stenoses following radiotherapy for neuroblastoma. AB - We describe the case of an 11-year-old girl with an abdominal neuroblastoma which was operated and intraoperatively irradiated nine years ago. After six years, she developed stenoses of the infrarenal abdominal aorta and both renal arteries. Initial treatment of the stenosed vessels comprised endovascular balloon dilatations and repeated stent-graft implantations, including drug eluting stents. However, severe in-stent stenoses occurred during follow-up and the girl developed acute renal failure. Open surgery was performed with two extra-anatomic bypasses, a thoracic-to-abdominal aortic bypass and a left iliac-to-renal bypass, on an urgent basis. The postoperative course was uneventful and the patient was discharged home two weeks after the operation with full recovery of renal function. We conclude that endovascular stent-graft placement in children can only be a palliative treatment due to outgrowing of the stent-graft and the potential risk of re-stenosis, especially after a history of irradiation. Vascular surgery with placement of extra-anatomic bypasses will provide a definite treatment. PMID- 19126554 TI - OCT4 expression in human non-small cell lung cancer: implications for therapeutic intervention. AB - Here we investigate the expression of OCT4 human lung adenocarcinoma and bronchioloalveolar carcinoma (BAC) tumor biopsies and tumor-derived primary cell cultures. OCT4 has been detected in several human tumors suggesting a potentially critical role in tumorigenesis. We assessed the presence of OCT4 in clinical tumor samples of both adenocarcinoma and BAC at the cellular and transcriptional levels, respectively. Furthermore, we evaluated tumor-derived cell cultures for potential differences in OCT4 expression. Immunohistochemical analysis depicted OCT4 in 2 of 8 adenocarcinoma tumor samples and 3 of 5 BAC tumor samples, with no apparent difference in the degree of expression among the sections examined. These results were validated by transcript analysis. Flow cytometric assessment of 11 adenocarcinoma-derived cell cultures and 3 BAC-derived cell cultures revealed significantly higher OCT4 expression in adenocarcinoma tumors compared to their normal counterparts. This, however, was not observed in the BAC cultures. Comparative studies of OCT4 in adenocarcinoma and BAC tumor cell cultures demonstrated a dramatically higher expression in the former. The expression of OCT4 may represent a specific and effective target for therapeutic intervention in adenocarcinoma and BAC. In addition, the aberrant expression and distribution of OCT4 may indicate important parameters concerning the differences between adenocarcinoma and BAC. PMID- 19126555 TI - Surgical treatment for patients with solitary metastasis in the mediastinal lymph node from renal cell carcinoma. AB - We performed surgical treatment on two patients, each with a solitary metastasis in a mediastinal lymph node from a renal cell carcinoma (RCC). The first case was a 58-year-old male with a chief complaint of chest discomfort due to pretracheal mediastinal lymph node (#3) swelling. He had undergone a right nephrectomy for RCC 13 years previously. Because of difficulty in establishing the diagnosis, a mini-thoracotomy was performed, and this lymphadenopathy was judged to be metastasis from the RCC. The pretracheal lymph nodes were completely resected, and he has experienced no recurrence for two years postoperatively. The second case was a 60-year-old female who had undergone a left nephrectomy for RCC two years previously. Because of the Botallo's lymph node (#5) swelling, a mini thoracotomy was performed. This swollen lymph node was resected, and it was finally diagnosed to be metastasis from the RCC. Unfortunately, the tumor recurred in the mediastinal lymph nodes with multiple lung metastases five years later. A solitary metastasis in a mediastinal lymph node from a RCC is an unusual event, particularly in the absence of lung metastasis. The diagnostic and clinicopathological problems associated with this unique disease are herein discussed. PMID- 19126556 TI - Prospective randomized evaluation of stentless vs. stented aortic biologic prosthetic valves in the elderly at five years. AB - OBJECTIVES: Randomized trials comparing stentless to stented bioprostheses for aortic valve replacement in elderly are scarce. The aim of this study was early and mid-term evaluation of these bioprostheses, with regards to clinical outcome and hemodynamic performance. METHODS: Between September 1999 and January 2001, 40 patients with aortic stenosis, over the age of 75 years, were randomly assigned to receive either the stented Perimount (n=20) or the stentless Prima Plus (n=20) bioprosthesis. Clinical outcomes, left ventricular mass regression, effective orifice area, ejection fraction and mean gradients were evaluated at discharge, six months, one year and five years after surgery. RESULTS: At five years, there were 5/20 (25%) deaths in the stentless group and 6/20 (30%) deaths in the stented group (all non-valve-related). There was one case of endocarditis in each group, early postoperatively. Overall, a significant decrease in left ventricular mass was found five years postoperatively. However, there was no significant difference in the rate and completeness of LV-mass regression between the groups (LV mass index 114+/-34.1 vs. 120+/-27.2). Furthermore, hemodynamic performance of the valves (mean gradient of 9.9+/-4.8 mmHg vs. 10.2+/-4.2 mmHg) did not differ significantly between the groups. CONCLUSIONS: At five years, stentless valves were not superior to the stented valves, with regards to hemodynamic performance, regression of left ventricular mass and clinical outcome. PMID- 19126557 TI - Evaluation of oxygen radical absorbance capacity in kampo medicine. AB - Antioxidant capacity of food has come to be shown in terms of oxygen radical absorbance capacity (ORAC) mainly on vegetables or fruit. However, the evaluation of Kampo in terms of ORAC has not yet been accomplished. It is important that such an investigation is also conducted for Kampo medicine. We measured the ORAC value of almost all the available Kampo formulas used in the Japanese National health insurance system and examined the ORAC value both for the daily prescription, and also the crude herb ingredients. The ORAC value of Kampo medicine ranged 4.65-5913 units/day. The ORAC value was high in Kampo formulas including Rhei Rhizoma, and was relatively high in Kampo formulas including anti inflammatory herbs other than Rhei Rhizoma. The ORAC value was also high in Kampo formulas including crude herbs that have relaxation effects. The ORAC value of a crude herb would seem to not be fixed but be dependent on combination with other crude herbs from the comparison of different herbs added to the basic Kampo medicine. These results suggest variability and complexity of the antioxidant capacity of Kampo medicine within the similar range of food. On the other hand, investigation of the compound changes of various crude herbs with ORAC may lead to the elucidation of the action mechanism of Kampo medicine. PMID- 19126559 TI - Comparative effectiveness of tumour necrosis factor alpha inhibitors in combination with either methotrexate or leflunomide. AB - OBJECTIVE: The objective of this study was to compare the effectiveness of a combination of tumour necrosis factor alpha (TNFalpha) inhibitors with either methotrexate or leflunomide in the treatment of patients with rheumatoid arthritis in a real-world setting. METHODS: Data from 1769 outpatients enrolled in the German biologics register RABBIT who were treated with one of the TNFalpha inhibitors adalimumab, etanercept, or infliximab in combination with either methotrexate (n = 1375) or leflunomide (n = 394) were included in the analysis. Clinical status including disease activity as well as treatment data were documented by the treating rheumatologist at baseline and at 3, 6, 12, 18, 24, 30 and 36 months of follow-up. RESULTS: Patients treated with a combination of biologics with leflunomide had significantly higher baseline disease activity than those treated with methotrexate. The highest disease activity was found for patients treated with the combination infliximab/leflunomide. After 36 months, the discontinuation rates were 46.3%, 51.3% and 61.5% for combinations of etanercept, adalimumab and infliximab with methotrexate and 53.4%, 63.1% and 67.1% for combinations with leflunomide, respectively. European League Against Rheumatism response rates after 24 months ranged from 74% to 81% for combinations with methotrexate and 72% to 81% for combinations with leflunomide. CONCLUSION: The current clinical practice is to use methotrexate as a first choice for the combination with TNFalpha antagonists. In a number of patients methotrexate has to be replaced by another disease-modifying antirheumatic drug. Our data support the view that leflunomide is a useful alternative if methotrexate is contraindicated. PMID- 19126560 TI - Periodontal disease in patients with ankylosing spondylitis. AB - OBJECTIVE: Ankylosing spondylitis (AS) and periodontal disease (PD) are characterised by dysregulation of the host inflammatory response, resulting in soft and hard connective tissue destruction. AS has been related to other inflammatory diseases, however, there is a paucity of data on whether AS is associated with inflammatory PD. METHODS: The association between AS and PD was examined in 48 patients with AS and 48 healthy controls, matched for age and gender. AS was diagnosed according to the modified New York criteria. Periodontal examination included probing pocket depth (PPD), clinical attachment loss (CAL), plaque index (PI) and bleeding on probing (BOP). Potential risk factors of PD such as smoking, low education, alcohol consumption, body mass index (BMI), as well as chronic diseases associated with PD and AS were assessed through questionnaires. RESULTS: In stepwise logistic regression, including AS status, age, gender, education, smoking, alcohol consumption and BMI, only AS status, age and education remained significant predictors of PD. Patients with AS had significant 6.81-fold increased odds (95% CI 1.96 to 23.67) of PD (defined as mean attachment loss >3 mm) compared to controls. The strength of the association was attenuated but remained statistically significant after further adjustment for plaque accumulation (odds ratio (OR) 5.48, 95% CI 1.37 to 22.00). CONCLUSIONS: The present study shows that patients with AS have a significantly higher risk of PD, strongly suggesting the need for close collaboration between rheumatologists, periodontists and dental hygienists when treating patients with AS. PMID- 19126561 TI - Lupus-specific health outcome measure for US patients: the LupusQoL-US version. AB - BACKGROUND: Patient-reported outcomes are valuable for the management of chronic diseases like systematic lupus erythematosus (SLE), but no measures have been validated for use in US-based patients with SLE. OBJECTIVES: To adapt and assess the validity and reliability of an SLE-specific quality of life (QoL) measure developed in the United Kingdom, the LupusQoL, for use in US-based patients with SLE. METHODS: Debriefing interviews of subjects with SLE guided the language modifications of the tool. The LupusQoL-US, SF-36 and EQ5D were administered. Internal consistency (ICR) and test-retest (TRT) reliability, convergent and discriminative validity were examined. Factor analyses were performed. RESULTS: The mean (SD) age of the 185 subjects with SLE was 42.5 (12.9) years. ICR and TRT of the eight domains ranged from 0.85 to 0.94 and 0.68 to 0.92, respectively. Related domains on the SF-36 correlated with the LupusQoL domains (physical health and physical function r = 0.73, physical health and role physical r = 0.57, emotional health and mental health r = 0.72, emotional health and role emotional r = 0.48, pain and bodily pain r = 0.66, fatigue and vitality r = 0.70, planning and social functioning r = 0.58). Most LupusQoL-US domains could discriminate between subjects with varied disease activity and damage. Principal component analysis disclosed five factors in the US version, with physical function, pain and planning items loading on one factor. CONCLUSIONS: These data provide evidence to support the psychometric properties of the LupusQoL-US, suggesting its utility as an assessment tool for patients with SLE in the USA. PMID- 19126562 TI - Working conditions predict incidence of long-term spells of sick leave due to depression: results from the Belstress I prospective study. AB - BACKGROUND: During the last few years, a high incidence of sick leave due to depression has been reported, resulting in important economic and social impacts. Only a limited number of studies investigating the influence of psychosocial working conditions on sick leave have been prospective and have utilised a valid methodology, while none have studied sick leave due to depression. In this study, the impact of adverse psychosocial working conditions is analysed on the risk for long-term sick leave due to depression. METHODS: This study resulted from the large-scale Belstress I study on the relationship between perceived job stress and health problems. Subjects were Belgian employees selected from 11 large companies (n = 9396). Using a longitudinal design, the association between the three Karasek stress dimensions (job control, psychological demand, and social support) was explored, separately and combined according to the demand-control and demand-control-support models and the incidence of long-term sick leave for depression as diagnosed by the family physician. RESULTS: After adjusting for age, occupational categories, living situation, and baseline depression score, 'passive jobs' (OR 2.67; 95% CI 1.15 to 6.19) and 'high strain' jobs (OR 3.23; 95% CI 1.40 to 7.43) predicted sick leave due to depression at follow-up in men. Job control predicted sick leave due to depression in both men (OR 2.43; 95% CI 1.27 to 4.66) and women (OR 2.21; 95% CI 1.05 to 4.68). CONCLUSIONS: This study provides evidence that the psychosocial working environment influences long-term sick leave due to depression. Efforts to improve skill discretion and decision authority at work could help prevent depression. PMID- 19126563 TI - Serrated and non-serrated polyps of the colorectum: their prevalence in an unselected case series and correlation of BRAF mutation analysis with the diagnosis of sessile serrated adenoma. AB - AIMS: To determine the prevalence of colorectal polyps of different types in an unselected population, and to correlate the morphological diagnoses with BRAF mutation analysis. METHODS: Cases of colorectal polyps diagnosed at endoscopy were retrieved from the files of Southern.IML Pathology. All slides were reviewed and the lesions classified histologically. A diagnosis of sessile serrated adenoma was made even if the characteristic features were present only focally. If there was more than one polyp of a particular type in any patient, one lesion was chosen at random so that the results represent the number of patients with each type of polyp rather than the total number of polyps. A proportion of the polyps was subjected to BRAF mutation analysis. RESULTS: A total of 1479 patients were identified. Non-serrated ("conventional") adenomas were found in 964 patients (65%), hyperplastic polyps in 437 (30%), sessile serrated adenomas in 57 (3.9%), traditional serrated adenomas in 11 (0.7%) and mixed hyperplastic adenomatous polyps in 10 (0.7%). BRAF V600E mutation analysis was performed in 148 selected cases; mutations were found in 44/49 (90%) of lesions diagnosed as sessile serrated adenoma, in 10/34 (29%) of hyperplastic polyps of microvesicular type, in 4/11 (36%) of traditional serrated adenomas, in 10/10 (100%) of mixed hyperplastic adenomatous polyps, and in 2/42 (5%) of "conventional" adenomas. CONCLUSIONS: Sessile serrated adenomas are encountered commonly in routine endoscopy practice. The histological diagnosis correlates strongly with the presence of BRAF mutation. PMID- 19126564 TI - Histological scoring of fibrosis and activity in HIV-chronic hepatitis B related liver disease: performance of the METAVIR score assessed on virtual slides. AB - BACKGROUND: The METAVIR score, which is the most widely used score in France, was specifically elaborated and evaluated in chronic hepatitis C and has never been validated in HIV-hepatitis virus B (HBV) co-infected patients. AIMS: To validate the use of the METAVIR scoring system for activity and fibrosis on liver biopsies in co-infected HIV-HBV patients. METHODS: METAVIR scoring for activity and fibrosis was first conducted on both original and virtual slides by one pathologist for comparison. Then 55 biopsies turned into virtual slides were scored by three pathologists independently. RESULTS: The scoring comparison between virtual slides and glass slides showed an almost perfect agreement for fibrosis (weighted kappa (kappa(w)) 0.8) and a substantial agreement for activity (kappa(w) 0.68). The inter-observer agreement on virtual slides was moderate to almost perfect (kappa(w) 0.52 to 0.84) for fibrosis and was dependent on the pair of pathologists considered. The best agreement was obtained in scoring advanced fibrosis and cirrhosis versus significant fibrosis versus no or mild fibrosis (kappa(w) 0.70 to 0.84). The agreement for cirrhosis was rated moderate to substantial (kappa(w) 0.54 to 0.79). Agreement for activity was substantial (kappa(w) 0.66 to 0.8). CONCLUSIONS: This study validates the use of virtual slide technology to assess fibrosis and activity on liver biopsies. It also validates the use of the METAVIR score in co-infected HIV-HBV patients and illustrates the challenges in establishing the histological diagnosis of cirrhosis in the HIV-HBV context. PMID- 19126565 TI - CD30-positive diffuse large B-cell lymphoma with microvillous features: so-called microvillous lymphoma. AB - A case of CD30-positive microvillous lymphoma (MVL) in an 87-year-old man who was encountered generalised lymphadenopathy is presented. Histopathologically, the tumour showed a morphological mimic of anaplastic large cell lymphoma (ALCL) with sinusoidal growth pattern. Immunohistochemically (IHC), the tumour cells were CD30(+), CD20(+), CD45(+), BCL-2(+), BCL-6(+), MUM1(+), Ki-67(+), CD45RO(-), CD3( ), CD10(-), CD15(-), CD56(-), EMA(-), TIA-1(-) and ALK(-). Flow cytometry confirmed the IHC. In situ hybridisation for Epstein-Barr virus RNA was negative. Electron microscopically, the tumour cells were similar to large transformed lymphocytes and had circumferentially profuse microvillous projections resembling those of epithelial mesothelioma cells. In conclusion, CD30-positive MVLs are indistinguishable from ALCLs that have ultrastructural microvillous projections by morphology alone. However, the lack of EMA, TIA-1 and ALK expression in this MVL case facilitated a definite distinction from ALCLs. The results of a panel of three markers (CD10(-), Bcl-6(+) and MUM1(+)) suggested that the present case of CD30-positive MVLs has an activated non-germinal centre B-cell origin. PMID- 19126566 TI - VEGF-A and i-NOS expression are prognostic factors in serous epithelial ovarian carcinomas after complete surgical resection. AB - AIMS: Clinical stage at the time of diagnosis and achievement of complete macroscopic resection during initial surgery are key factors determining the outcome of ovarian cancer. However, prediction of outcome lacks accuracy and more reliable prognostic factors are required. Therefore, an analysis and evaluation of key angiogenic factors was carried out to determine their diagnostic and prognostic value in serous ovarian cancer. METHODS: Expression levels of vascular endothelial growth factor (VEGF)-A, hypoxia-inducible factor (HIF)1-alpha and inducible nitric oxide synthase (i-NOS) were analysed by immunohistochemistry in a homogenous group of 112 patients with serous adenocarcinoma of the ovary. Vascular density as an indicator of angiogenesis was assessed using the Chalkley eyepiece method after staining for CD34. The correlation of these data with survival and established prognostic factors such as histological grade, Federation of Gynecology and Obstetrics (FIGO) stage, and residual tumour after surgery, was evaluated. Survival analyses, multivariate analyses and correlation tests were performed. RESULTS: In the patient group with macroscopic complete tumour resection (R0) there was a significant correlation between VEGF-A and i NOS expression. Kaplan-Meier analysis further revealed improved progression-free survival for R0 patients with VEGF-A-positive and i-NOS-negative tumours. The predictive relevance of VEGF-A regarding progression-free survival was sustained in multivariate analysis using FIGO stage, grading and resection status as fixed variables. CONCLUSION: VEGF-A and i-NOS are prognostic markers for clinical outcome in serous ovarian cancer patients with macroscopic complete tumour resection (R0). Hence, pre-therapeutic assessment of VEGF-A as predictive factor for an antiangiogenic therapy might be of clinical value. PMID- 19126567 TI - The role of immunohistochemistry in the differential diagnosis of papillary lesions of the breast. AB - Papillary lesions of the breast represent a heterogeneous group with differing biological behaviour. Correct diagnosis is crucial but may be difficult, as many benign and malignant papillary lesions have similar appearances. Immunohistochemistry plays a useful role in their differentiation. Myoepithelial markers can help in differentiating papilloma from papillary carcinoma, as the former usually shows a continuous layer of myoepithelial cells. In intracystic papillary carcinoma, there is controversy as to the presence of a complete myoepithelial cell layer around these lesions. p63 is the marker of choice as the staining is nuclear, cross-reactivity is minimal, and sensitivity is high. Papilloma may frequently be complicated by superimposed different types of epithelial hyperplasia, which range from usual to atypical or even ductal carcinoma in situ, and they many be morphologically similar. Basal cytokeratins (CKs) are useful to differentiate these entities; as usual hyperplasia is positive for basal CKs with a mosaic staining pattern. CK5/6 is probably the best marker. Neuroendocrine markers (chromogranin A and synaptophysin) may be positive in papillary carcinoma, particularly in the solid type, and there may be some overlap with the ductal carcinoma in situ with spindle cells or endocrine ductal carcinoma in situ. A panel of CK5/6, p63 and neuroendocrine markers can be useful in the diagnostic investigation of problematic papillary lesions of the breast. As the experience with these markers remains rather limited, it is too early to recommend basing treatment choices on these marker studies alone. Complete removal of lesion is probably still the treatment of choice. PMID- 19126568 TI - RNA interference screening demystified. AB - Genetic screens, where the effects of modifying gene function on cell behaviour are assessed in a systematic fashion, have for some time provided useful information to those interested in disease pathogenesis and treatment. Genetic screens exploiting the phenomenon of RNA interference (RNAi) are now becoming commonplace. This article explains the different RNAi screen formats and describes some of the applications of RNAi screening that may be pertinent to the research pathologist. PMID- 19126569 TI - A pilot open label, single dose trial of fenobam in adults with fragile X syndrome. AB - OBJECTIVE: A pilot open label, single dose trial of fenobam, an mGluR5 antagonist, was conducted to provide an initial evaluation of safety and pharmacokinetics in adult males and females with fragile X syndrome (FXS). METHODS: Twelve subjects, recruited from two fragile X clinics, received a single oral dose of 50-150 mg of fenobam. Blood for pharmacokinetic testing, vital signs and side effect screening was obtained at baseline and numerous time points for 6 h after dosing. Outcome measures included prepulse inhibition (PPI) and a continuous performance test (CPT) obtained before and after dosing to explore the effects of fenobam on core phenotypic measures of sensory gating, attention and inhibition. RESULTS: There were no significant adverse reactions to fenobam administration. Pharmacokinetic analysis showed that fenobam concentrations were dose dependent but variable, with mean (SEM) peak values of 39.7 (18.4) ng/ml at 180 min after the 150 mg dose. PPI met a response criterion of an improvement of at least 20% over baseline in 6 of 12 individuals (4/6 males and 2/6 females). The CPT did not display improvement with treatment due to ceiling effects. CONCLUSIONS: Clinically significant adverse effects were not identified in this study of single dose fenobam across the range of dosages utilised. The positive effects seen in animal models of FXS treated with fenobam or other mGluR5 antagonists, the apparent lack of clinically significant adverse effects, and the potential beneficial clinical effects seen in this pilot trial support further study of the compound in adults with FXS. PMID- 19126570 TI - 19q13.11 deletion syndrome: a novel clinically recognisable genetic condition identified by array comparative genomic hybridisation. AB - BACKGROUND: Deletions of chromosome 19 have rarely been reported, with the exception of some patients with deletion 19q13.2 and Blackfan-Diamond syndrome due to haploinsufficiency of the RPS19 gene. Such a paucity of patients might be due to the difficulty in detecting a small rearrangement on this chromosome that lacks a distinct banding pattern. Array comparative genomic hybridisation (CGH) has become a powerful tool for the detection of microdeletions and microduplications at high resolution in patients with syndromic mental retardation. METHODS AND RESULTS: Using array CGH, this study identified three interstitial overlapping 19q13.11 deletions, defining a minimal critical region of 2.87 Mb, associated with a clinically recognisable syndrome. The three patients share several major features including: pre- and postnatal growth retardation with slender habitus, severe postnatal feeding difficulties, microcephaly, hypospadias, signs of ectodermal dysplasia, and cutis aplasia over the posterior occiput. Interestingly, these clinical features have also been described in a previously reported patient with a 19q12q13.1 deletion. No recurrent breakpoints were identified in our patients, suggesting that no-allelic homologous recombination mechanism is not involved in these rearrangements. CONCLUSIONS: Based on these results, the authors suggest that this chromosomal abnormality may represent a novel clinically recognisable microdeletion syndrome caused by haploinsufficiency of dosage sensitive genes in the 19q13.11 region. PMID- 19126571 TI - Detection of Neisseria gonorrhoeae and Chlamydia trachomatis in pharyngeal and rectal specimens using the BD Probetec ET system, the Gen-Probe Aptima Combo 2 assay and culture. AB - OBJECTIVES: This study compared the sensitivity and specificity of culture and two nucleic acid amplification tests (NAATs): the BD Probetec ET system (PT) and the Aptima Combo 2 (AC2) in detecting Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) in pharyngeal and rectal specimens. METHODS: Male subjects were prospectively recruited at an MSM clinic in Toronto, Canada. Pharyngeal and rectal specimens were obtained for GC and CT culture, PT and AC2. Urine was also obtained for PT. A true positive was defined as: (1) positive culture, (2) positive PT and AC2 at the same site or (3) a single positive NAAT and detection of the same organism by any method at another site. RESULTS: 248 subjects were recruited. The prevalence of pharyngeal GC was 8.1%, rectal GC 11.7%, pharyngeal CT 2.0% and rectal CT 7.7%. The sensitivity of culture for pharyngeal GC and CT was 0%; 41.4% for rectal GC and 21.1% for rectal CT. The sensitivity of PT for pharyngeal GC, rectal GC, pharyngeal CT and rectal CT was 95.0%, 93.1%, 80.0% and 94.7%, respectively. The sensitivity of AC2 was 95.0% for pharyngeal GC and 100% at all other sites. Specificity was consistently above 98%. CONCLUSIONS: PT and AC2 detected GC and CT with superior sensitivity compared to culture. They detected 73 pharyngeal or rectal GC and CT infections compared to 16 by culture, using a rigorous gold standard. NAATs should be the method of choice for the detection of GC and CT in extragenital sites in men who have sex with men. PMID- 19126572 TI - Understanding social dysfunction in the behavioural variant of frontotemporal dementia: the role of emotion and sarcasm processing. AB - Social interaction is profoundly affected in the behavioural form of frontotemporal dementia (bvFTD) yet there are few means of objectively assessing this. Diagnosis of bvFTD is based on informant report, however a number of individuals with a clinical profile consistent with the disease have no imaging abnormality and seem to remain stable, with doubt about the presence of underlying neurodegenerative pathology. We aimed to quantify aspects of the behavioural disorder and link it to the underlying level of atrophy in socially relevant brain regions. We tested individuals with either bvFTD (N = 26) or Alzheimer's disease (N = 9) and 16 controls using The Awareness of Social Inference Test (TASIT) to assess their ability to identify emotion and sarcasm in video vignettes. A subset of bvFTD patients (N = 21) and controls (N = 12) were scanned using MRI within 6 months of assessment. There was marked impairment in the ability of bvFTD patients whose scans showed abnormalities to recognize sarcastic, but not sincere statements. Their capacity to interpret negative emotion was also impaired, and this appeared to be a major factor underlying the deficit in sarcasm recognition. Clinically diagnosed bvFTD patients whose scans were normal, Alzheimer's disease patients and controls had no difficulty in appreciating both types of statement. In a multivariate imaging analysis it was shown that the sarcasm (and emotion recognition) deficit was dependent on a circuit involving the lateral orbitofrontal cortex, insula, amygdala and temporal pole, particularly on the right. Performance on a more global test of cognitive function, the Addenbrooke's Cognitive Examination did not have a unique association with these regions. The TASIT is an objective test of social dysfunction in bvFTD which indexes the frontotemporal volume loss in bvFTD patients and provides an objective measure for separating behavioural patients who are likely to decline from those who may remain stable. These results provide additional evidence for the role of the orbitofrontal cortex and related structures in the processing of socially relevant signals, particularly those where negative emotion recognition is important. PMID- 19126573 TI - Testing at home--the screening of the future? PMID- 19126574 TI - A divide-and-conquer approach to analyze underdetermined biochemical models. AB - MOTIVATION: To obtain meaningful predictions from dynamic computational models, their uncertain parameter values need to be estimated from experimental data. Due to the usually large number of parameters compared to the available measurement data, these estimation problems are often underdetermined meaning that the solution is a multidimensional space. In this case, the challenge is yet to obtain a sound system understanding despite non-identifiable parameter values, e.g. through identifying those parameters that most sensitively determine the model's behavior. RESULTS: Here, we present the so-called divide-and-conquer approach--a strategy to analyze underdetermined biochemical models. The approach draws on steady state omics measurement data and exploits a decomposition of the global estimation problem into independent subproblems. The solutions to these subproblems are joined to the complete space of global optima, which can be easily analyzed. We derive the conditions at which the decomposition occurs, outline strategies to fulfill these conditions and--using an example model- illustrate how the approach uncovers the most important parameters and suggests targeted experiments without knowing the exact parameter values. PMID- 19126575 TI - The Flannotator--a gene and protein expression annotation tool for Drosophila melanogaster. AB - SUMMARY: Understanding developmental processes and building towards integrative systems biology require detailed knowledge of the spatio-temporal expression of genes and proteins. We have developed a software package for collecting, storing and searching the annotation of protein or gene expression patterns in Drosophila melanogaster. Using standard Drosophila anatomy and Gene Ontologies, the system can readily capture expression patterns at any stage of development and in all recognized tissue types as well as details of sub-cellular localization. The web based system allows multiple groups to work in collaboration and share images and annotation. AVAILABILITY: http://www.flannotator.org.uk/. PMID- 19126576 TI - Host-parasite relations of bacteria and phages can be unveiled by oligostickiness, a measure of relaxed sequence similarity. AB - MOTIVATION: The recent metagenome analysis has been producing a large number of host-unassigned viruses. Although assigning viruses to their hosts is basically important not only for virology but also for prevention of epidemic, it has been a laborious and difficult task to date. The only effective method for this purpose has been to find them in a same microscopic view. Now, we tried a computational approach based on genome sequences of bacteria and phages, introducing a physicochemical parameter, SOSS (set of oligostickiness similarity score) derived from oligostickiness, a measure of binding affinity of oligonucleotides to template DNA. RESULTS: We could confirm host-parasite relationships of bacteria and their phages by SOSS analysis: all phages tested (25 species) had a remarkably higher SOSS value with its host than with unrelated bacteria. Interestingly, according to SOSS values, lysogenic phages such as lambda phage (host: Escherichia coli) or SPP1 (host: Bacillus subtilis) have distinctively higher similarity with its host than its non-lysogenic (excretive or virulent) ones such as fd and T4 (host: E.coli) or phages gamma and PZA (host: B.subtilis). This finding is very promising for assigning host-unknown viruses to its host. We also investigated the relationship in codon usage frequency or G+C content of genomes to interpret the phenomenon revealed by SOSS analysis, obtaining evidences which support the hypothesis that higher SOSS values resulted from the cohabitation in the same environment which may cause the common biased mutation. Thus, lysogenic phages which stay inside longer resemble the host. PMID- 19126577 TI - BioCichlid: central dogma-based 3D visualization system of time-course microarray data on a hierarchical biological network. AB - SUMMARY: BioCichlid is a 3D visualization system of time-course microarray data on molecular networks, aiming at interpretation of gene expression data by transcriptional relationships based on the central dogma with physical and genetic interactions. BioCichlid visualizes both physical (protein) and genetic (regulatory) network layers, and provides animation of time-course gene expression data on the genetic network layer. Transcriptional regulations are represented to bridge the physical network (transcription factors) and genetic network (regulated genes) layers, thus integrating promoter analysis into the pathway mapping. BioCichlid enhances the interpretation of microarray data and allows for revealing the underlying mechanisms causing differential gene expressions. AVAILABILITY: BioCichlid is freely available and can be accessed at http://newton.tmd.ac.jp/. Source codes for both biocichlid server and client are also available. PMID- 19126581 TI - Variants of the arachidonate 5-lipoxygenase-activating protein (ALOX5AP) gene and risk of stroke: a HuGE gene-disease association review and meta-analysis. AB - Variants of the arachidonate 5-lipoxygenase-activating protein (ALOX5AP) gene have been implicated as a risk factor for stroke. However, genetic association studies that have examined the association between ALOX5AP gene variants (HapA haplotype, HapB haplotype, and SG polymorphisms) and stroke have produced conflicting results. Therefore, the authors performed a meta-analysis of all studies with ALOX5AP genotyping (5,194 stroke cases and 4,566 controls). The meta analysis showed significant heterogeneity among studies (P(Q) = 0.03, I(2) = 63%) and a nonsignificant association between the HapA haplotype (SG13S25G-SG13S114T SG13S89G-SG13S32A) and stroke risk (random-effects (RE) odds ratio (OR) = 1.13, 95% confidence interval (CI): 0.88, 1.45). Regarding the HapB haplotype (SG13S377A-SG13S114A-SG13S41A-SG13S35G), there was no association with stroke risk (RE OR = 1.03, 95% CI: 0.77, 1.37). The SG13S114, SG13S89, SG13S25, SG13S32, SG13S35, and SG13S42 polymorphisms were not associated with stroke. The SG13S106 and SG13S377 polymorphisms revealed evidence of marginal association (RE OR = 1.23 (95% CI: 1.03, 1.46) and RE OR = 1.25 (95% CI: 1.04, 1.50), respectively). However, cumulative meta-analysis for the HapA haplotype showed a downward trend of odds ratios over time, and recursive cumulative meta-analysis indicated insufficient evidence for claiming or denying an association. Tests for bias revealed no evidence of biases. Rigorous genetic association studies investigating gene-gene-environment interactions may generate more conclusive claims about the genetics of stroke. PMID- 19126579 TI - The role of IGF-1R in pediatric malignancies. AB - The insulin-like growth factor (IGF) family consists of ligands (IGF-I, IGF-II, insulin), several receptors (including IGF-1R), and six binding proteins (IGFBP-1 through IGFBP-6). Members of this family regulate key cellular activities and they also play an important role in the development and progression of both adult and childhood cancers. Binding of a ligand to the receptor leads to its activation, followed by signal transduction along several pathways. In some childhood malignancies, IGF-1R can be activated by endocrine, autocrine, or paracrine mechanisms. Although mutations in IGF-1R have not been identified, this signaling pathway is upregulated in many childhood cancers. These findings have led to the development of a host of IGF-1R signaling modulators that are currently being tested in clinical trials. This review explores the role of IGF 1R in a range of childhood malignancies. PMID- 19126580 TI - Tuning the relative affinities for activating and repressing operators of a temporally regulated restriction-modification system. AB - Most type II restriction-modification (R-M) systems produce separate endonuclease (REase) and methyltransferase (MTase) proteins. After R-M genes enter a new cell, MTase activity must appear before REase or the host chromosome will be cleaved. Temporal control of these genes thus has life-or-death consequences. PvuII and some other R-M systems delay endonuclease expression by cotranscribing the REase gene with the upstream gene for an autogenous activator/repressor (C protein). C.PvuII was previously shown to have low levels early, but positive feedback later boosts transcription of the C and REase genes. The MTase is expressed without delay, and protects the host DNA. C.PvuII binds to two sites upstream of its gene: O(L), associated with activation, and O(R), associated with repression. Even when symmetry elements of each operator are made identical, C.PvuII binds preferentially to O(L). In this study, the intra-operator spacers are shown to modulate relative C.PvuII affinity. In light of a recently reported C.Esp1396I DNA co-crystal structure, in vitro and in vivo effects of altering O(L) and O(R) spacers were determined. The results suggest that the GACTnnnAGTC consensus is the primary determinant of C.PvuII binding affinity, with intra-operator spacers playing a fine-tuning role that affects mobility of this R-M system. PMID- 19126582 TI - Relation of dietary glycemic index, glycemic load, and fiber and whole-grain intakes during puberty to the concurrent development of percent body fat and body mass index. AB - The authors prospectively examined whether change in dietary glycemic index (GI), glycemic load (GL), fiber intake, or whole-grain intake during puberty is associated with concurrent change in percentage of body fat (%BF) or body mass index (BMI; weight (kg)/height)(2). Linear mixed-effects regression analyses were performed in 215 participants from the Dortmund Nutritional and Anthropometric Longitudinally Designed (DONALD) Study (Dortmund, Germany) who possessed weighed 3-day dietary records and anthropometric data at puberty onset (defined by age at takeoff) and over the subsequent 4 years (1988-2007). Neither changes in dietary GI, GL, fiber intake, nor whole-grain intake were associated with concurrent changes in %BF throughout puberty (change in %BF: -0.03 (standard error (SE), 0.11) per standard deviation (SD) increase in GI (P = 0.8); -0.01 (SE, 0.11) per SD increase in GL (P = 0.9); 0.02 (SE, 0.14) per SD increase in fiber intake (P = 0.9); and 0.09 (SE, 0.13) per SD increase in whole-grain intake (P = 0.5)). Similarly, no concurrent associations were observed between these dietary factors and BMI SD scores. Associations of dietary GI with %BF and BMI SD score differed between overweight and normal-weight adolescents (for concurrent association, P for interaction was 0.03 for %BF and 0.08 for BMI SD score). Dietary GI, GL, and fiber and whole-grain intakes in healthy, free-living adolescents do not appear to be relevant to the development of %BF or BMI during puberty. PMID- 19126583 TI - Methodological issues in analyzing time trends in biologic fertility: protection bias. AB - One method of assessing biologic fertility is to measure time to pregnancy (TTP). Accidental pregnancies do not generate a valid TTP value and lead to nonrandom missing data if couples experiencing accidental pregnancies are more fertile than the general population. If factors affecting the rate of accidental pregnancies, such as availability of effective contraception and induced abortion, vary over time, then the result may be protection bias in the estimates of fertility time trends. Six European data sets were analyzed to investigate whether evidence of protection bias exists in TTP studies of fertility trends in Europe over the past 50 years. Couples experiencing accidental pregnancies tended to be more fertile than the general population. However, trends in accidental pregnancy rates were inconsistent across countries and were insufficient to produce substantial bias in fertility trends in simulated data. Where protection bias is suspected, the authors demonstrate use of 2 multiple imputation methods to generate realizations for the missing TTP values for accidental pregnancies. Simulation studies show that both methods successfully reduce or eliminate protection bias. The authors also demonstrate that standard sensitivity analyses for dealing with accidental pregnancies provide an upper bound on the extent of any bias. PMID- 19126584 TI - An outcome-based approach for the creation of fetal growth standards: do singletons and twins need separate standards? AB - Contemporary fetal growth standards are created by using theoretical properties (percentiles) of birth weight (for gestational age) distributions. The authors used a clinically relevant, outcome-based methodology to determine if separate fetal growth standards are required for singletons and twins. All singleton and twin livebirths between 36 and 42 weeks' gestation in the United States (1995 2002) were included, after exclusions for missing information and other factors (n = 17,811,922). A birth weight range was identified, at each gestational age, over which serious neonatal morbidity and neonatal mortality rates were lowest. Among singleton males at 40 weeks, serious neonatal morbidity/mortality rates were lowest between 3,012 g (95% confidence interval (CI): 3,008, 3,018) and 3,978 g (95% CI: 3,976, 3,980). The low end of this optimal birth weight range for females was 37 g (95% CI: 21, 53) less. The low optimal birth weight was 152 g (95% CI: 121, 183) less for twins compared with singletons. No differences were observed in low optimal birth weight by period (1999-2002 vs. 1995-1998), but small differences were observed for maternal education, race, parity, age, and smoking status. Patterns of birth weight-specific serious neonatal morbidity/neonatal mortality support the need for plurality-specific fetal growth standards. PMID- 19126585 TI - Occupational exposures and asthma in health-care workers: comparison of self reports with a workplace-specific job exposure matrix. AB - The authors compared self-reported occupational exposures with a workplace specific job exposure matrix (JEM) in a 2004 survey of Texas health-care professionals (n = 3,650), by asthma status. Sensitivity, specificity, chance corrected (kappa) and chance-independent (phi) agreement, and associations of self-reported exposures with asthma were compared with those for the JEM. Among asthmatics, the median sensitivity of self-reported exposures was 74% (range, 53 90); specificity was 64% (range, 27-74). For nonasthmatics, median sensitivity was 67% (range, 40-88) and specificity was 70% (range, 33-82). Sensitivity was higher among asthmatics for exposures involving perceptible odors. Specificity was higher among nonasthmatics for instrument cleaning and exposure to adhesives/solvents. Asthmatics showed better agreement with the JEM for patient care-related cleaning (phi = 0.51 vs. 0.40); there was little difference for other exposures. In all cases, confidence intervals overlapped. Prevalence ratios were higher with self-reported exposures than with the JEM; differences were greatest for cleaning products, adhesives/solvents, and gases/vapors. However, confidence intervals overlapped with those obtained using the JEM. In asthma studies, differential reporting bias by health status should be taken into consideration. Findings favor using externally developed methods of exposure classification, although information gleaned from examining distributions of exposure self-reports, particularly among nondiseased persons, can provide useful information for improving the reliability of exposure ascertainment. PMID- 19126586 TI - Hardy-Weinberg equilibrium testing of biological ascertainment for Mendelian randomization studies. AB - Mendelian randomization (MR) permits causal inference between exposures and a disease. It can be compared with randomized controlled trials. Whereas in a randomized controlled trial the randomization occurs at entry into the trial, in MR the randomization occurs during gamete formation and conception. Several factors, including time since conception and sampling variation, are relevant to the interpretation of an MR test. Particularly important is consideration of the "missingness" of genotypes that can be originated by chance, genotyping errors, or clinical ascertainment. Testing for Hardy-Weinberg equilibrium (HWE) is a genetic approach that permits evaluation of missingness. In this paper, the authors demonstrate evidence of nonconformity with HWE in real data. They also perform simulations to characterize the sensitivity of HWE tests to missingness. Unresolved missingness could lead to a false rejection of causality in an MR investigation of trait-disease association. These results indicate that large scale studies, very high quality genotyping data, and detailed knowledge of the life-course genetics of the alleles/genotypes studied will largely mitigate this risk. The authors also present a Web program (http://www.oege.org/software/hwe-mr calc.shtml) for estimating possible missingness and an approach to evaluating missingness under different genetic models. PMID- 19126587 TI - Cost-effectiveness of acupuncture in women and men with allergic rhinitis: a randomized controlled study in usual care. AB - To assess quality of life and cost-effectiveness of additional acupuncture treatment for allergic rhinitis, patients were randomly allocated to 2 groups; both received usual care, but one group received an additional 10 acupuncture sessions. Quality of life (according to the SF-36 Health Survey), and direct and indirect costs, were assessed at baseline and after 3 months, and the incremental cost-effectiveness ratio of acupuncture treatment was calculated. This German study (December 2000-June 2004) involved 981 patients (64% women, mean age 40.9 years (standard deviation, 11.2); 36% men, mean age 43.2 years (standard deviation, 13.0)). At 3 months, quality of life was higher in the acupuncture group than in the control group (mean Physical Component Score 51.99 (standard error (SE), 0.33) vs. 48.25 (SE, 0.33), P < 0.001; mean Mental Component Score 48.55 (SE, 0.42) vs. 45.35 (SE, 0.42), respectively, P < 0.001). Overall costs in the acupuncture group were significantly higher than those in the control group (Euro (euro; 1 euro = US $1.27) 763, 95% confidence interval: 683, 844 vs. 332 euro, 95% confidence interval: 252, 412; mean difference 432 euro, 95% confidence interval: 318, 545). The incremental cost-effectiveness ratio was 17,377 euro per quality-adjusted life year (women, 10,155 euro; men, 44,871 euro) and was robust in sensitivity analyses. Acupuncture, supplementary to routine care, was beneficial and, according to international benchmarks, cost-effective. However, because of the study design, it remains unclear whether the effects are acupuncture specific. PMID- 19126588 TI - Environmental tobacco smoke and lung cancer among Chinese nonsmoking males: might adenocarcinoma be the culprit? AB - No studies have specifically reported the association of lung adenocarcinoma with environmental tobacco smoke (ETS) exposure among nonsmoking males. The objective of this study was to examine the exposure-response relation between ETS exposure and lung cancer among nonsmoking males. In particular, the association with adenocarcinoma of the lung was studied. This is a population-based, case-referent study in Hong Kong during 2004-2006. A total of 132 Chinese male nonsmokers with newly diagnosed primary lung cancer and 536 nonsmoking community referents were interviewed about ETS exposures from the household and/or workplace, including ever ETS exposure, sources of exposure, number of smoking cohabitants/coworkers, and smoker-years. Univariate logistic regression analyses showed a weak association between all lung cancers and ever ETS exposure from the household and/or workplace (odds ratio (OR) = 1.11, 95% confidence interval (CI): 0.74, 1.67), but an increased risk was restricted to adenocarcinoma (OR = 1.68, 95% CI: 1.00, 2.38). After adjustment for family cancer history and other confounders, excess risk (OR = 1.62, 95% CI: 0.91, 2.88) still persisted for adenocarcinoma, although it was no longer statistically significant. Exposure-response relations for adenocarcinoma were found with increasing levels of all ETS indices when exposures from the household and workplaces were combined. The consistent exposure-response relations between ETS exposures and adenocarcinoma suggested a probable causal link, which would have to be confirmed by future larger studies. PMID- 19126589 TI - Indoor charcoal smoke and acute respiratory infections in young children in the Dominican Republic. AB - The authors investigated the effect of charcoal smoke exposure on risks of acute upper and lower respiratory infection (AURI and ALRI) among children under age 18 months in Santo Domingo, Dominican Republic (1991-1992). Children living in households using charcoal for cooking (exposed, n = 201) were age-matched to children living in households using propane gas (nonexposed, n = 214) and were followed for 1 year or until 2 years of age. Fuel use and new episodes of AURI and ALRI were ascertained biweekly through interviews and medical examinations. Household indoor-air concentration of respirable particulate matter (RPM) was measured in a sample of follow-up visits. Incidences of AURI and ALRI were 4.4 and 1.4 episodes/child-year, respectively. After adjustment for other risk factors, exposed children had no significant increase in risk of AURI but were 1.56 times (95% confidence interval: 1.23, 1.97) more likely to develop ALRI. RPM concentrations were higher in charcoal-using households (27.9 microg/m(3) vs. 17.6 microg/m(3)), and ALRI risk increased with RPM exposure (10-microg/m(3) increment: odds ratio = 1.17, 95% confidence interval: 1.02, 1.34). Exposure to charcoal smoke increases the risk of ALRI in young children, an effect that is probably mediated by RPM. Reducing charcoal smoke exposure may lower the burden of ALRI among children in this population. PMID- 19126591 TI - Prospective study of physical activity and lung cancer by histologic type in current, former, and never smokers. AB - Increased physical activity has been associated with decreased lung cancer risk. However, no previous investigation has examined physical activity in relation to lung cancer histologic types by smoking status. The authors investigated these relations in the National Institutes of Health-AARP Diet and Health Study among 501,148 men and women aged 50-71 years at baseline in 1995-1996. During follow-up to 2003, 6,745 lung carcinomas occurred (14.8% small cell, 40.3% adenocarcinoma, 19.7% squamous cell, 6.1% undifferentiated large cell, 7.2% non-small cell not otherwise specified, and 11.8% carcinoma not otherwise specified). Among former smokers, the multivariate relative risks of small cell, adenocarcinoma, squamous cell, and undifferentiated large cell carcinomas comparing the highest with the lowest activity level (> or =5 times/week vs. inactive) were 0.93 (95% confidence interval (CI): 0.67, 1.28), 0.79 (95% CI: 0.67, 0.94), 0.73 (95% CI: 0.57, 0.93), and 0.61 (95% CI: 0.38, 0.98), respectively. Among current smokers, corresponding values were 0.77 (95% CI: 0.58, 1.02), 0.76 (95% CI: 0.61, 0.95), 0.85 (95% CI: 0.65, 1.11), and 1.10 (95% CI: 0.69, 1.78). In contrast, physical activity was unrelated to lung carcinoma among never smokers (P(interaction) between physical activity and smoking for total lung carcinomas = 0.002). The inverse findings among former and current smokers in combination with the null results for physical activity among never smokers may point toward residual confounding by cigarette smoking as an explanation for the relations observed. PMID- 19126590 TI - Long working hours and cognitive function: the Whitehall II Study. AB - This study examined the association between long working hours and cognitive function in middle age. Data were collected in 1997-1999 (baseline) and 2002-2004 (follow-up) from a prospective study of 2,214 British civil servants who were in full-time employment at baseline and had data on cognitive tests and covariates. A battery of cognitive tests (short-term memory, Alice Heim 4-I, Mill Hill vocabulary, phonemic fluency, and semantic fluency) were measured at baseline and at follow-up. Compared with working 40 hours per week at most, working more than 55 hours per week was associated with lower scores in the vocabulary test at both baseline and follow-up. Long working hours also predicted decline in performance on the reasoning test (Alice Heim 4-I). Similar results were obtained by using working hours as a continuous variable; the associations between working hours and cognitive function were robust to adjustments for several potential confounding factors including age, sex, marital status, education, occupation, income, physical diseases, psychosocial factors, sleep disturbances, and health risk behaviors. This study shows that long working hours may have a negative effect on cognitive performance in middle age. PMID- 19126592 TI - Estimating the impact of the discontinuation of medical interventions on health outcomes. AB - The same study designs are used to document the magnitude of the influence of currently used and of previously used treatments on health outcomes. However, because randomized trials rarely are utilized to study cessation of treatment, most of what we know about the experience of former users of a given treatment comes from cohort and case-control studies. These studies generally compare former users with never users or current users, but occasionally different groups of former users are compared according to how long ago the treatment was stopped. The experience of whole populations following relatively rapid and widespread discontinuation of a previously common treatment sometimes can serve as a secondary source of data on the consequences of discontinuation of therapy. PMID- 19126593 TI - Helix 8 of leukotriene B4 type-2 receptor is required for the folding to pass the quality control in the endoplasmic reticulum. AB - Many G protein-coupled receptors (GPCRs) possess a putative cytoplasmic helical domain, termed helix 8 (H8), at the proximal region of the C-terminal tail. However, the significance of this domain is not fully understood. Here, we demonstrate the requirement of H8 for the proper folding of GPCRs for passage through the quality control in the endoplasmic reticulum (ER). In the human leukotriene B(4) type-2 receptor (hBLT2), lack of H8 led to an accumulation of the receptor (hBLT2/DeltaH8) in the ER. Similar results were obtained in two representative human GPCRs, dopamine type-1 and lysophosphatidic acid type-2 receptors, which were engineered to lack H8. Treatment with the several ligands, which act as pharmacological chaperones, facilitated the surface expression of hBLT2/DeltaH8. The surface-trafficked hBLT2/DeltaH8 exhibited an agonist-evoked increase in Ca(2+), demonstrating that H8 is not critical for ligand binding and activation of coupled G proteins. Thus, these results suggest that the H8 region of hBLT2 plays an important role in transport-competent receptor folding. PMID- 19126594 TI - Crystal structure of human esterase D: a potential genetic marker of retinoblastoma. AB - Retinoblastoma (RB), a carcinoma of the retina, is caused by mutations in the long arm of chromosome 13, band 13q14. The esterase D (ESD) gene maps at a similar location as the RB gene locus and therefore serves as a potential marker for the prognosis of retinoblastoma. Because very little is known about the structure and function of ESD, we determined the 3-dimensional structure of the enzyme at 1.5 A resolution using X-ray crystallography. ESD shows a single domain with an alpha/beta-hydrolase fold. A number of insertions are observed in the canonical alpha/beta-hydrolase fold. The active site is located in a positively charged, shallow cleft on the surface lined by a number of aromatic residues. Superimposition studies helped identify the typical catalytic triad residues--Ser 153, His264, and Asp230--involved in catalysis. Mutagenesis of any of the catalytic triad residues to alanine abolished the enzyme activity. Backbone amides of Leu54 and Met150 are involved in the formation of the oxyanion hole. Interestingly, a M150A mutation increased the enzyme activity by 62%. The structure of human ESD determined in this study will aid the elucidation of the physiological role of the enzyme in the human body and will assist in the early diagnosis of retinoblastoma. PMID- 19126595 TI - Poor maternal nutrition followed by accelerated postnatal growth leads to telomere shortening and increased markers of cell senescence in rat islets. AB - Low birth weight (LBW) followed by accelerated postnatal growth is associated with increased risk of developing age-associated diseases such as type 2 diabetes. Gestational protein restriction in rats causes LBW, beta-cell dysfunction, and reduced longevity. These effects may be mediated by accelerated cellular aging. This study tested the hypothesis that LBW followed by rapid postnatal catch-up growth leads to islet telomere shortening through alterations in antioxidant defense capacity, stress/senescence marker proteins, and DNA repair mechanisms at the gene expression level. We used our rat model of gestational protein restriction (recuperated offspring) and control offspring. Southern blotting revealed shorter (P<0.001) islet telomeres in recuperated animals compared to controls. This was associated with increased expression of peroxiredoxin 1 (P<0.05), peroxiredoxin 3 (P<0.01), and heme oxygenase-1 (HO-1) (P<0.05), which are up-regulated under stress conditions. MnSOD expression was significantly (P<0.05) decreased in recuperated offspring, suggesting partial impairment of mitochondrial antioxidant defenses. Markers of cellular senescence p21 and p16 were also increased (P<0.01 and P<0.05, respectively) in the recuperated group. We conclude that maternal diet influences expression of markers of cellular stress and telomere length in pancreatic islets. This may provide a mechanistic link between early nutrition and growth and type 2 diabetes. PMID- 19126596 TI - Essential role of Rac1 and Rac3 GTPases in neuronal development. AB - Rac GTPases are members of the Rho family regulating the actin cytoskeleton and implicated in neuronal development. Ubiquitous Rac1 and neuron-specific Rac3 GTPases are coexpressed in the developing mammalian brain. We used Cre-mediated conditional deletion of Rac1 in neurons combined with knockout of neuron-specific Rac3 to study the role of these GTPases in neural development. We found that lack of both genes causes motor behavioral defects, epilepsy, and premature death of mice. Deletion of either GTPase does not produce evident phenotypes. Double knockout mice show specific defects in the development of the hippocampus. Selective impairment of the dorsal hilus of double-knockout animals is associated with alteration in the formation of the hippocampal circuitry. Axonal pathways to and from the dorsal hilus are affected because of the deficit of hilar mossy cells. Moreover, analysis of Rac function in hippocampal cultures shows that spine formation is strongly hampered only in neurons lacking both Rac proteins. These findings show for the first time that both Rac1 and Rac3 are important for the development of the nervous system, wherein they play complementary roles during late stages of neuronal and brain development. PMID- 19126598 TI - Oxidative stress in developmental origins of disease: teratogenesis, neurodevelopmental deficits, and cancer. AB - In the developing embryo and fetus, endogenous or xenobiotic-enhanced formation of reactive oxygen species (ROS) like hydroxyl radicals may adversely alter development by oxidatively damaging cellular lipids, proteins and DNA, and/or by altering signal transduction. The postnatal consequences may include an array of birth defects (teratogenesis), postnatal functional deficits, and diseases. In animal models, the adverse developmental consequences of in utero exposure to agents like thalidomide, methamphetamine, phenytoin, benzo[a]pyrene, and ionizing radiation can be modulated by altering pathways that control the embryonic ROS balance, including enzymes that bioactivate endogenous substrates and xenobiotics to free radical intermediates, antioxidative enzymes that detoxify ROS, and enzymes that repair oxidative DNA damage. ROS-mediated signaling via Ras, nuclear factor kappa B and related transducers also may contribute to altered development. Embryopathies can be reduced by free radical spin trapping agents and antioxidants, and enhanced by glutathione depletion. Further modulatory approaches to evaluate such mechanisms in vivo and/or in embryo culture have included the use of knockout mice, transgenic knock-ins and mutant deficient mice with altered enzyme activities, as well as antisense oligonucleotides, protein therapy with antioxidative enzymes, dietary depletion of essential cofactors and chemical enzyme inhibitors. In a few cases, measures anticipated to be protective have conversely enhanced the risk of adverse developmental outcomes, indicating the complexity of development and need for caution in testing therapeutic strategies in humans. A better understanding of the developmental effects of ROS may provide insights for risk assessment and the reduction of adverse postnatal consequences. PMID- 19126597 TI - Modification of surfactant protein D by reactive oxygen-nitrogen intermediates is accompanied by loss of aggregating activity, in vitro and in vivo. AB - Surfactant protein D (SP-D) is an important effector of innate immunity. We have previously shown that SP-D accumulates at sites of acute bacterial infection and neutrophil infiltration, a setting associated with the release of reactive species such as peroxynitrite. Incubation of native SP-D or trimeric SP-D lectin domains (NCRDs) with peroxynitrite resulted in nitration and nondisulfide cross linking. Modifications were blocked by peroxynitrite scavengers or pH inactivation of peroxynitrite, and mass spectroscopy confirmed nitration of conserved tyrosine residues within the C-terminal neck and lectin domains. Mutant NCRDs lacking one or more of the tyrosines allowed us to demonstrate preferential nitration of Tyr314 and the formation of Tyr228-dependent cross-links. Although there was no effect of peroxynitrite or tyrosine mutations on lectin activity, incubation of SP-D dodecamers or murine lavage with peroxynitrite decreased the SP-D-dependent aggregation of lipopolysaccharide-coated beads, supporting our hypothesis that defective aggregation results from abnormal cross-linking. We also observed nitration, cross-linking of SP-D, and a significant decrease in SP D-dependent aggregating activity in the lavage of mice acutely exposed to nitrogen dioxide. Thus, modification of SP-D by reactive oxygen-nitrogen species could contribute to alterations in the structure and function of SP-D at sites of inflammation in vivo. PMID- 19126599 TI - Cadmium toxicity toward autophagy through ROS-activated GSK-3beta in mesangial cells. AB - We previously demonstrated that cadmium (Cd) is able to induce autophagic cell death through a calcium-extracellular signal-regulated kinase pathway. Here, the object of this study is to investigate the role of glycogen synthase kinase-3beta (GSK-3beta) in the induction of autophagy. After treatment with Cd, MES-13 mesangial cells were determined to have undergone autophagy based on the formation of acidic vesicular organelles and autophagosomes as well as on the processing of microtubule-associated protein 1 light chain 3, using flow cytometry with acridine orange staining, electron microscopy, and immunoblot, respectively. Use of the GSK-3beta inhibitor SB 216763 or the small interfering RNA technique to knockdown the expression of GSK-3beta resulted in a decrease of Cd-induced autophagy. In contrast, overexpression of GSK-3beta by transient transfection potentiated Cd toxicity toward the mesangial cells, suggesting that GSK-3beta plays a crucial role in regulating Cd-induced autophagy. Moreover, a decrease of the phosphorylated level at Ser9 of GSK-3beta was observed by immunoblot after treatment with Cd, indicating GSK-3beta was activated by Cd. This phenomenon was reversed by the reactive oxygen species (ROS) scavenger N acetylcysteine (NAC), demonstrated that ROS might activate GSK-3beta. In fact, intracellular hydrogen peroxide (H(2)O(2)) was 2.6-fold elevated after 3 h of exposure to Cd. Both Cd-induced ROS bursts and autophagy were reduced by NAC and vitamin E. In summary, this study demonstrated that, in MES-13 mesangial cells, Cd-induced autophagy was mediated through the ROS-GSK-3beta signaling pathway. PMID- 19126600 TI - Potential neurotoxicity of ketamine in the developing rat brain. AB - Ketamine, an N-methyl-D-aspartate (NMDA) receptor ion channel blocker, is a widely used anesthetic recently reported to enhance neuronal death in developing rodents and nonhuman primates. This study evaluated dose-response and time-course effects of ketamine, levels of ketamine in plasma and brain, and the relationship between altered NMDA receptor expression and ketamine-induced neuronal cell death during development. Postnatal day 7 rats were administered 5, 10, or 20 mg/kg ketamine using single or multiple injections (subcutaneously) at 2-h intervals, and the potential neurotoxic effects were examined 6 h after the last injection. No significant neurotoxic effects were detected in layers II or III of the frontal cortex of rats administered one, three, or six injections of 5 or 10 mg/kg ketamine. However, in rats administered six injections of 20 mg/kg ketamine, a significant increase in the number of caspase-3- and Fluoro-Jade C positive neuronal cells was observed in the frontal cortex. Electron microscopic observations showed typical nuclear condensation and fragmentation indicating enhanced apoptotic characteristics. Increased cell death was also apparent in other brain regions. In addition, apoptosis occurred after plasma and brain levels of ketamine had returned to baseline levels. In situ hybridization also showed a remarkable increase in mRNA signals for the NMDA NR1 subunit in the frontal cortex. These data demonstrate that ketamine administration results in a dose-related and exposure-time dependent increase in neuronal cell death during development. Ketamine-induced cell death appears to be apoptotic in nature and closely associated with enhanced NMDA receptor subunit mRNA expression. PMID- 19126601 TI - Tirofiban increases soluble guanylate cyclase in rat vascular walls: pharmacological and pathophysiological consequences. AB - AIMS: Our aim was to evaluate whether tirofiban, which mimics the structure of arginine-glycine-aspartic acid (RGD) peptides, up-regulates soluble guanylate cyclase beta1 subunit (sGC-beta1) expression in vascular smooth muscle cells (VSMCs) and in aorta from rats, and to investigate the pharmacological and pathophysiological consequences of this up-regulation. METHODS AND RESULTS: Wistar, Wistar Kyoto, and spontaneously hypertensive rats (SHRs) were used. sGC beta1 content was assessed by immunoblotting. Arterial pressure was recorded using a tail-cuff sphygmomanometer. Sodium nitroprusside (SNP) and isosorbide dinitrate (IDN) were used as nitric oxide (NO) donors. Tirofiban increased the sGC-beta1 content in VSMCs and in aortic walls from rats after 6 h of treatment. Rats treated with tirofiban experienced a more pronounced decrease in their arterial pressure after acute SNP treatment than vehicle-treated rats. Isolated rat aortic rings incubated with tirofiban showed a higher relaxing response to SNP than control rings as well as an increased sGC-beta1 content and SNP-induced cyclic guanosine monophosphate synthesis. Animals receiving IDN for 1 week showed decreased sGC-beta1 in aortic walls and did not respond to SNP treatment with changes in arterial pressure. Tirofiban restored the decreased sGC-beta1 content in IDN-treated rats and promoted a decreased arterial pressure in response to SNP administration. SHRs showed reduced sGC-beta1 levels, and tirofiban increased these levels and led to a higher response to SNP. CONCLUSION: Tirofiban increased the sGC-beta1 content in contractile cells and aortic walls of rats, enhancing the response to SNP and reversing the NO donor tachyphylaxis. PMID- 19126603 TI - Regulation's threat to research. Over-regulation, Russian style. PMID- 19126604 TI - Amputation by text. See also amputation by internet. PMID- 19126602 TI - Expression of CYP1A1 and CYP1B1 in human endothelial cells: regulation by fluid shear stress. AB - AIMS: CYP1A1 and CYP1B1, members of the cytochrome P450 protein family, are regulated by fluid shear stress. This study describes the effects of duration, magnitude and pattern of shear stress on CYP1A1 and CYP1B1 expressions in human endothelial cells, towards the goal of understanding the role(s) of these genes in pro-atherogenic or anti-atherogenic endothelial cell functions. METHODS AND RESULTS: We investigated CYP1A1 and CYP1B1 expressions under different durations, levels, and patterns of shear stress. CYP1A1 and CYP1B1 mRNA, protein, and enzymatic activity were maximally up-regulated at > or =24 h of arterial levels of shear stress (15-25 dynes/cm2). Expression of both genes was significantly attenuated by reversing shear stress when compared with 15 dynes/cm2 steady shear stress. Small interfering RNA knockdown of CYP1A1 resulted in significantly reduced CYP1B1 and thrombospondin-1 expression, genes regulated by the aryl hydrocarbon receptor (AhR). Immunostaining of human coronary arteries showed constitutive CYP1A1 and CYP1B1 protein expressions in endothelial cells. Immunostaining of mouse aorta showed nuclear localization of AhR and increased expression of CYP1A1 in the descending thoracic aorta, whereas reduced nuclear localization of AhR and attenuated CYP1A1 expression were observed in the lesser curvature of the aortic arch. CONCLUSION: CYP1A1 and CYP1B1 gene and protein expressions vary with time, magnitude, and pattern of shear stress. Increased CYP1A1 gene expression modulates AhR-regulated genes. Based on our in vitro reversing flow data and in vivo immunostained mouse aorta, we suggest that increased expression of both genes reflects an anti-atherogenic endothelial cell phenotype. PMID- 19126606 TI - The blame game. PMID- 19126607 TI - Controversy sparked in Switzerland after doctor who used unproved, unlicensed therapy is cleared of non-treatment conviction. PMID- 19126608 TI - Case of Baby P. Let's go back to basics in child protection. PMID- 19126609 TI - Early detection of melanoma is key, so let's teach it. PMID- 19126610 TI - Duty in reporting stab wounds. PMID- 19126611 TI - Regulation's threat to research. Regulation of research. PMID- 19126612 TI - Trading laws and health foods. Lip service legislation. PMID- 19126613 TI - Irritable bowel syndrome. Why pills more than skills? PMID- 19126614 TI - The problem of insoluble fibre in irritable bowel syndrome. PMID- 19126615 TI - Irritable bowel syndrome. Don't forget amitriptyline. PMID- 19126616 TI - Coordination improves outcomes in malignant cord compression. PMID- 19126618 TI - US university psychiatrist loses chairmanship over drug company payments. PMID- 19126619 TI - English government launches initiative to cut obesity. PMID- 19126624 TI - Presence of tungsten-containing fibers in tungsten refining and manufacturing processes. AB - In tungsten refining and manufacturing processes, a series of tungsten oxides are typically formed as intermediates in the production of tungsten powder. The present study was conducted to characterize airborne tungsten-containing fiber dimensions, elemental composition and concentrations in the US tungsten refining and manufacturing industry. During the course of normal employee work activities, seven personal breathing zone and 62 area air samples were collected and analyzed using National Institute for Occupational Safety and Health (NIOSH) fiber sampling and counting methods to determine dimensions, composition and airborne concentrations of fibers. Mixed models were used to identify relationships between potential determinants and airborne fiber concentrations. Results from transmission electron microscopy analyses indicated that airborne fibers with length >0.5 microm, diameter >0.01 microm and aspect ratios > or =3:1 were present on 35 of the 69 air samples collected. Overall, the airborne fibers detected had a geometric mean length approximately 3 microm and diameter approximately 0.3 microm. Ninety-seven percent of the airborne fibers identified were in the thoracic fraction (i.e. aerodynamic diameter < or = 10 microm). Energy dispersive X-ray spectrometry results indicated that airborne fibers prior to the carburization process consisted primarily of tungsten and oxygen, with other elements being detected in trace quantities. Based on NIOSH fiber counting 'B' rules (length > 5 microm, diameter < 3 microm and aspect ratio > or = 5:1), airborne fiber concentrations ranged from below the limit of detection to 0.085 fibers cm(-3), with calcining being associated with the highest airborne concentrations. The mixed model procedure indicated that process temperature had a marginally significant relationship to airborne fiber concentration. This finding was expected since heated processes such as calcining created the highest airborne fiber concentrations. The finding of airborne tungsten-containing fibers in this occupational setting needs to be confirmed in similar settings and demonstrates the need to obtain information on the durability and associated health effects of these fibers. PMID- 19126625 TI - Effects of dehydroepiandrosterone therapy on pubic hair growth and psychological well-being in adolescent girls and young women with central adrenal insufficiency: a double-blind, randomized, placebo-controlled phase III trial. AB - CONTEXT AND OBJECTIVE: The efficacy of oral dehydroepiandrosterone (DHEA) in the treatment of atrichia pubis and psychological distress in young females with central adrenal insufficiency is unknown. Our study aimed to evaluate this therapy. DESIGN AND PATIENTS: A total of 23 young females (mean age 18 yr, range 13-25) was enrolled in a double-blind randomized placebo-controlled trial. Inclusion criteria were ACTH deficiency plus two or more additional pituitary deficiencies, serum DHEA less than 400 ng/ml, and pubertal stage more than B2. Exclusion criteria were cerebral radiation with more than 30 Gy, tumor remission less than 1 yr, amaurosis, hypothalamic obesity, psychiatric disorders, and unstable hormone medication. INTERVENTION: Patients were randomized to placebo (n = 12) or 25 mg HPLC-purified DHEA/d (n = 11) orally for 12 months after stratification into a nontumor (n = 7) and a tumor group (n = 16). MAIN OUTCOME MEASURES: Clinical scoring of pubic hair stage was performed at 0, 6, and 12 months (primary endpoint), and psychometrical evaluation (Symptom Check-List-90-R and the Centre for Epidemiological Studies-Depression Scale) at 0 and 12 months (secondary endpoint). Androgen levels and safety parameters were measured at 0, 6, and 12 months; 24-h androgen urinary excretion rates were calculated at 0 and 12 months. RESULTS: In the placebo group, four patients dropped out because of recurrence of craniopharyngioma, manifestation of type 1 diabetes, and change of residence (n = 2); in the DHEA group, one patient dropped out because of recurrent anxiety attacks. DHEA substitution resulted in normalization of DHEA sulfate and androstanediol glucuronide morning serum levels 2 h after drug intake (P < 0.006), and of its 24 h urinary metabolite levels (P < 0.0001), placebo had no effect. Morning serum levels of androstenedione increased in the DHEA group (P < 0.02) but did not normalize. The DHEA group exhibited significant progress in pubic hair growth from Tanner stage I-III to II-V (mean: +1.5 stages), whereas the placebo group did not (relative risk 0.138; 95% confidence interval 0.021 0.914; P = 0.0046). Importantly, eight of the 10 Symptom Check-List-90-R scores, including those for depression, anxiety, and interpersonal sensitivity, and the global severity index improved in the DHEA group in comparison to the placebo group (P < 0.048). DHEA was well tolerated. CONCLUSIONS: In adolescent girls with central adrenal insufficiency, daily replacement with 25 mg DHEA orally is beneficial: atrichia pubis vanishes, and psychological well-being improves significantly. PMID- 19126626 TI - Adipokines, inflammation, and visceral adiposity across the menopausal transition: a prospective study. AB - CONTEXT: Postmenopausal women have greater visceral adiposity compared with premenopausal women. Adipokines are associated with increased adiposity, insulin resistance, and atherosclerosis. OBJECTIVE: The objective of the study was to assess changes in adipokines and inflammatory markers through the menopausal transition and correlate them with changes in visceral adiposity. DESIGN AND SETTING: This was a prospective cohort study of women through the menopausal transition conducted at the University of Washington. PARTICIPANTS: Sixty-nine healthy women were followed up longitudinally from premenopausal (aged 45-55 yr) to postmenopausal status (aged 49-60 yr). OUTCOME: On premenopausal and postmenopausal visits, fasting blood was drawn for adiponectin, leptin, serum amyloid A (SAA), C-reactive protein (CRP), monocyte-chemotactic protein-1, tissue plasminogen activator antigen (tPA), IL-6, and TNF-alpha. Body composition measures were assessed by body mass index, whole-body dual x-ray absorptiometry scan, and computed tomography scan of the abdomen at the lumbar 4-5 level. RESULTS: Women had a statistically significant increase in SAA, tPA, monocyte chemotactic protein-1, and adiponectin between the two measurement occasions (P = 0.04, P = 0.02, P = 0.001, and P < 0.001, respectively). The increase in intraabdominal fat was correlated positively with the change in SAA (r = 0.31, P = 0.02), CRP (r = 0.56, P < 0.001), tPA (r = 0.40, P = 0.002), and leptin (r = 0.41, P = 0.002) and negatively correlated with the change in adiponectin (r = 0.37, P = 0.005). After adjustment for change in sc abdominal fat, the correlation between change in CRP, tPA, leptin, and adiponectin remained significantly associated with change in intraabdominal fat. CONCLUSIONS: Women going through the menopausal transition have deleterious changes in inflammatory markers and adipokines that correlate with increased visceral adiposity. PMID- 19126627 TI - Genetic modifiers play a substantial role in diabetes complicating cystic fibrosis. AB - CONTEXT: Insulin-requiring diabetes affects 7-15% of teens and young adults, and more than 25% of older adults with cystic fibrosis (CF). Pancreatic exocrine disease caused by CF transmembrane conductance regulator (CFTR) dysfunction underlies the high rate of diabetes in CF patients; however, only a subset develops this complication, indicating that other factors are necessary. OBJECTIVE: Our objective was to estimate the relative contribution of genetic and nongenetic modifiers to the development of diabetes in CF. DESIGN/PATIENTS: This was a twin and sibling study involving 1366 individuals at 109 centers in the CF Twin and Sibling Study, from which were derived 68 monozygous twin pairs, 23 dizygous twin pairs, and 588 sibling pairs, all with CF. MAIN OUTCOME MEASURE: Chronic, insulin-requiring diabetes in the setting of CF, as established using longitudinal clinical and biochemical data, was studied. RESULTS: About 9% of this predominantly pediatric population (mean age = 15.8 yr) had diabetes. Key independent risk factors identified by regression modeling included having a twin or sibling with CF and diabetes, increasing age, pancreatic exocrine insufficiency or two mutations causing severe CFTR dysfunction, decreased lung function or decreased body mass index, and longer duration of glucocorticoid treatment. The concordance rate for diabetes was substantially higher in monozygous twins (0.73) than in dizygous twins and siblings with CF (0.18; P = 0.002). Heritability was estimated as near one (95% confidence interval 0.42 1.0). CONCLUSIONS: Diabetes is a frequent complication of CF that is associated with worse outcomes. Although a nongenetic factor (steroid treatment) contributes to risk, genetic modifiers (i.e. genes other than CFTR) are the primary cause of diabetes in CF. PMID- 19126628 TI - High frequency of and factors associated with thyroid hormone over-replacement and under-replacement in men and women aged 65 and over. AB - CONTEXT: Thyroid hormone use is common in older populations, but the frequency of over- or under-replacement is debated. OBJECTIVE: We sought to describe the frequency of and factors associated with thyroid hormone over- or under replacement in a population of older men and women. DESIGN: Participants were 3678 U.S. community dwelling individuals aged 65 yr or older enrolled in the Cardiovascular Health Study who had thyroid function tests in 1989-1990. Thyroid hormone users (n = 339) were identified and classified into low TSH (<0.45 mU/liter), euthyroid (0.45-4.5 mU/liter), and high TSH (>4.5 mU/liter). RESULTS: Of the 339 thyroid hormone users, 41% had a low TSH, 16% had a high TSH, and 43% were in the euthyroid range. In multivariable analyses, lower weight (P < 0.001) was independently associated with low TSH status. For every 10 kg lower weight, the likelihood of having low TSH increased by 65% [odd ratio (OR) 1.65; 95% confidence interval (CI) 1.31-2.07]. Those with renal insufficiency were less likely to have low TSH levels (P = 0.02). The presence of diabetes was independently associated with having low (OR 3.35; 95% CI 1.46-7.65) and high TSH levels (OR 2.66; 95% CI 1.14-6.21). CONCLUSIONS: There is a very high prevalence of thyroid function testing abnormalities in older people taking thyroid hormone preparations, particularly in those of low weight or with diabetes. Because of potential adverse cardiovascular and skeletal effects from over-replacement, older people represent a key population for increased TSH monitoring on therapy. PMID- 19126629 TI - A novel mutation in the LIM homeobox 3 gene is responsible for combined pituitary hormone deficiency, hearing impairment, and vertebral malformations. AB - CONTEXT: The LIM homeobox 3 (LHX3) LIM-homeodomain transcription factor gene, found in both man and mouse, is required for development of the pituitary and motor neurons, and is also expressed in the auditory system. OBJECTIVE: The objective of this study was to determine the cause of, and further explore, the phenotype in six patients (aged 6 months to 22 yr) with combined pituitary hormone deficiency (CPHD), restricted neck rotation, scoliosis, and congenital hearing impairment. Three of the patients also have mild autistic-like behavior. DESIGN: Because patients with CPHD and restricted neck rotation have previously been shown to have mutations in the LHX3 gene, a candidate gene approach was applied, and the gene was sequenced. Neck anatomy was explored by computed tomography and magnetic resonance imaging, including three-dimensional reformatting. RESULTS: A novel, recessive, splice-acceptor site mutation was found. The predicted protein encoded by the mutated gene lacks the homeodomain and carboxyl terminus of the normal, functional protein. Genealogical studies revealed a common gene source for all six families dating back to the 17th century. Anatomical abnormalities in the occipito-atlantoaxial joints in combination with a basilar impression of the dens axis were found in all patients assessed. CONCLUSIONS: This study extends both the mutations known to be responsible for LHX3-associated syndromes and their possible phenotypical consequences. Previously reported traits include CPHD and restricted neck rotation; patients examined in the present study also show a severe hearing defect. In addition, the existence of cervical vertebral malformations are revealed, responsible for the rigid neck and the development of scoliosis. PMID- 19126630 TI - Extensive and largely reversible ischemic cerebral infarctions in a prepubertal child with hypertension and Cushing disease. PMID- 19126631 TI - Testicular effects of isolated luteinizing hormone deficiency and reversal by long-term human chorionic gonadotropin treatment. PMID- 19126632 TI - Polycystic ovary syndrome, inflammation, and statins: do we have the right target? PMID- 19126633 TI - Can a brief psychological intervention prevent anxiety or depressive disorders in cancer patients? A randomised controlled trial. AB - BACKGROUND: We tested whether a brief psychological intervention could prevent anxiety or depressive disorders among newly diagnosed cancer patients. PATIENTS AND METHODS: Patients free of anxiety or depressive disorder were randomised to receive immediate intervention (start of cancer treatment), delayed intervention (8 weeks after starting treatment) or usual care. They were stratified according to risk of developing anxiety or depressive disorders. Primary outcome was measured using a standardised psychiatric interview to detect any anxiety or depressive disorder at 6 and 12 months following the cancer diagnosis. Analyses used conditional odds logistic regression models adjusting for age, gender, concerns and past history to compare outcome of all intervention patients with usual care. RESULTS: A total of 465 patients were recruited. In all, 313 (79%) of the 397 well enough to be interviewed completed the study. At 12 months, there was no difference between the groups receiving the intervention and usual care [odds ratio (OR) = 0.69, 95% confidence interval (CI) 0.41-1.17, P = 0.17]. In high-risk patients, those who received the intervention were less likely to develop an anxiety or depressive disorder compared with those who received usual care (OR = 0.54, 95% CI 0.29-1.00, P = 0.050). In low-risk patients, there was no difference (OR = 1.50, 95% CI 0.51-4.43, P = 0.47). CONCLUSION: A brief intervention, delivered by nonspecialists, promoted adjustment among newly diagnosed cancer patients at high risk of developing anxiety or depressive disorders. PMID- 19126634 TI - Toward a terminology for functional recovery in schizophrenia: is functional remission a viable concept? AB - Recovery in schizophrenia is receiving increasing attention. Part of the increased focus is based on the recent working criteria for clinical remission in schizophrenia and the realization that many people with schizophrenia meet these criteria for remission. In this article, we consider whether functional disability can also be evaluated in a "remission" model. In so doing, we evaluate the concept of clinical remission, evaluate the possibility of remission of other generally stable features of schizophrenia such as negative symptoms, and make some heuristic terminological recommendations. We also propose a "level and breadth" model for the definition of functional remission and examine some of the alternate influences that could produce suboptimal everyday functioning, including effort, motivation, and societal barriers toward functional achievement. PMID- 19126635 TI - Domestication of cardamom (Elettaria cardamomum) in Western Ghats, India: divergence in productive traits and a shift in major pollinators. AB - BACKGROUND AND AIMS: Elettaria cardamomum, a highly priced spice, is native to the Western Ghats of South India. Wild populations still occur in isolated patches in their natural habitats; however, much of today's commercial product comes from cultivated sources. There is no information on domestication-related traits of this species; the main objective of this study was to compare wild and cultivated populations of cardamom in terms of vegetative and reproductive features in order to identify domestication syndromes and to examine whether the two populations have developed reproductive barriers. METHODS: Two wild populations and five cultivated plantations were used for the present study. Vegetative and floral traits, flowering phenology, pollination biology and breeding systems of wild and cultivated populations were compared. Effective pollinators amongst floral visitors were identified by confirming pollen transfer as well as by fruit set following their visit to virgin flowers. Manual pollinations were carried out in order to study the breeding systems of the two populations and reproductive barriers, if any, between them. KEY RESULTS: Several productive traits including the number of branches, number of inflorescences, and total number of flowers per clump, number of flowers that open each day, the duration of flowering, the length of the flower and the amount of nectar per flower are significantly greater in cultivated cardamom. The principal pollinators in wild cardamom are solitary bees, Megachile sp. and two species of Amegilla, whereas those in cultivated cardamom are the social bees Apis dorsata, A. cerana and Trigona iridipennis. Both the wild and cultivated populations are self-compatible and there are no reproductive barriers between the two populations. CONCLUSIONS: Domestication in cardamom has brought about significant changes in vegetative and reproductive traits and a shift in effective pollinators from native solitary bees to social bees. The shift in pollinators seems to be due to the availability of a large number of flowers for prolonged periods in cultivated cardamom that can attract and sustain social bees, rather than due to co-evolution of the flower and the pollinator. PMID- 19126636 TI - Phylogenetic analysis reveals a scattered distribution of autumn colours. AB - BACKGROUND AND AIMS: Leaf colour in autumn is rarely considered informative for taxonomy, but there is now growing interest in the evolution of autumn colours and different hypotheses are debated. Research efforts are hindered by the lack of basic information: the phylogenetic distribution of autumn colours. It is not known when and how autumn colours evolved. METHODS: Data are reported on the autumn colours of 2368 tree species belonging to 400 genera of the temperate regions of the world, and an analysis is made of their phylogenetic relationships in order to reconstruct the evolutionary origin of red and yellow in autumn leaves. KEY RESULTS: Red autumn colours are present in at least 290 species (70 genera), and evolved independently at least 25 times. Yellow is present independently from red in at least 378 species (97 genera) and evolved at least 28 times. CONCLUSIONS: The phylogenetic reconstruction suggests that autumn colours have been acquired and lost many times during evolution. This scattered distribution could be explained by hypotheses involving some kind of coevolutionary interaction or by hypotheses that rely on the need for photoprotection. PMID- 19126637 TI - A lucky cardiac shotgun? PMID- 19126638 TI - Temperature affects expression of symptoms induced by soybean mosaic virus in homozygous and heterozygous plants. AB - Seven strains (G1 to G7) of soybean mosaic virus (SMV) and 3 resistance loci (Rsv1, Rsv3, and Rsv4) have been identified in soybean. The interaction of SMV strains and host resistance genes results in resistant (symptomless), susceptible (mosaic), or necrotic (leaf and stem necrosis) reactions. The necrotic reaction may be gene dosage dependent and influenced by temperature. Using a set of soybean isolines and hybrids containing homozygous or heterozygous alleles of rsv, Rsv1, Rsv1-n, Rsv3, or Rsv4, this study has explored the relationship of SMV induced symptoms and resistance gene dosage at different temperatures. Results showed that SMV-inoculated plants carrying Rsv3 or Rsv4 were symptomless at both homozygous and heterozygous states at all temperature regimes. Threshold temperatures for symptoms changing from stem tip necrosis (STN) to mosaic were 30, 33, and 33 degrees C in G7-inoculated homozygous genotypes V94-3971(Rsv1) and PI 96983 (Rsv1) and G1-inoculated V262 (Rsv1-n), respectively. However, at the heterozygous state, threshold temperature was 30 degrees C in G7-inoculated V94 3971 x Essex F(1) for the symptom change from STN to mosaic, 31 degrees C in G7 inoculated Essex x PI 96983 F(1) from STN to mixture of necrosis and mosaic (N M), and 32 degrees C in G1-inoculated V262 x Essex F(1) from N-M to mosaic. Incomplete necrosis was observed in the heterozygous state in G1-inoculated V262 x Essex F(1) and G7-inoculated PI 96983 x Essex F(1) where necrotic and mosaic symptoms were mixed. High temperature (37 degrees C) tends to mask the expression of mosaic symptoms in both homozygous and heterozygous plants. STN expression in response to temperature was affected by resistance gene, gene dosage, host genetic background, and specific SMV strains. Thus, Rsv3 and Rsv4 are a better choice as source of genetic resistance for breeding SMV-resistant cultivars. PMID- 19126639 TI - MsatAllele_1.0: An R package to visualize the binning of microsatellite alleles. AB - MsatAllele is a computer package built on R to visualize and bin the raw microsatellite allele size distributions. The method is based on the creation of an R database with exported files from the open-source electropherogram peak reading program STRAND. Contrary to other binning programs, in this program, the bin limits are not fixed and are automatically defined by the data stored in the database. Data manipulation and graphical functions allow to 1) visualize raw allele size variation, allowing the detection of potential scoring errors, strange bin distributions, and unexpected spacing between the bins; 2) bin raw fragment sizes and write bin summary statistics for each locus; and 3) export genotype files with the resulting binned data. PMID- 19126640 TI - Sulfotransferase 1A1 haplotypes associated with oral squamous cell carcinoma susceptibility in male Taiwanese. AB - We have previously demonstrated that betel quid containing safrole induced DNA adducts are highly associated with the development of oral squamous cell carcinoma (OSCC) in Taiwan. Sulfotransferase (SULT) is essential for the formation of these adducts. To elucidate the effects of SULT1A1 haplotypes on OSCC susceptibility, 160 male OSCC cases and 218 age- and sex-matched controls were screened for single-nucleotide polymorphisms within the coding region of SULT1A1 by sequencing. We found that 445C>T (His149Tyr) and 507C>T polymorphisms were significantly associated with increased risk of OSCC. Based on the genotype analysis, haplotypes were constructed for 445C>T (His149Tyr), 507C>T, 600G>C and 638G>A (Arg213His) using GENECOUNTING software. After adjustment for age, cigarette smoking and betel quid chewing, we found that haplotype c containing 445C>T (His149Tyr), 507C>T or 600G>C but not 638G>A (Arg213His) variant was significantly associated with increased risk of OSCC (odds ratio, 3.24; 95% confidence interval, 1.57-6.68) when compared with the haplotype a (wild-type). We analyzed the activity in sulfonation of 2-naphthol and 1'-hydroxysafrole of recombinant His149Tyr (445C>T) variant, which led to 51 and 33% reduced activity, respectively; Arg213His (638G>A) variant led to 72 and 54% reduced activity, respectively, when compared with the wild-type. Taken together, haplotype analysis provides a novel evaluation of the SULT1A1 gene as a risk modifier on environmental carcinogen in OSCC and the association of SULT1A1 haplotypes with the risk of OSCC might be modified by betel quid chewing. PMID- 19126641 TI - Chemical genomics of cancer chemopreventive dithiolethiones. AB - 3H-1,2-dithiole-3-thione (D3T) and its analogues 4-methyl-5-pyrazinyl-3H-1,2 dithiole-3-thione (OLT) and 5-tert-butyl-3H-1,2-dithiole-3-thione (TBD) are chemopreventive agents that block or diminish early stages of carcinogenesis by inducing activities of detoxication enzymes. While OLT has been used in clinical trials, TBD has been shown to be more efficacious and possibly less toxic than OLT in animals. Here, we utilize a robust and high-resolution chemical genomics procedure to examine the pharmacological structure-activity relationships of these compounds in livers of male rats by microarray analyses. We identified 226 differentially expressed genes that were common to all treatments. Functional analysis identified the relation of these genes to glutathione metabolism and the nuclear factor, erythroid derived 2-related factor 2 pathway (Nrf2) that is known to regulate many of the protective actions of dithiolethiones. OLT and TBD were shown to have similar efficacies and both were weaker than D3T. In addition, we identified 40 genes whose responses were common to OLT and TBD, yet distinct from D3T. As inhibition of cytochrome P450 (CYP) has been associated with the effects of OLT on CYP expression, we determined the half maximal inhibitory concentration (IC(50)) values for inhibition of CYP1A2. The rank order of inhibitor potency was OLT >> TBD >> D3T, with IC(50) values estimated as 0.2, 12.8 and >100 microM, respectively. Functional analysis revealed that OLT and TBD, in addition to their effects on CYP, modulate liver lipid metabolism, especially fatty acids. Together, these findings provide new insight into the actions of clinically relevant and lead dithiolethione analogues. PMID- 19126642 TI - Epstein-Barr virus nuclear antigen 2 disrupts mitotic checkpoint and causes chromosomal instability. AB - The Epstein-Barr virus nuclear antigen 2 (EBNA2) plays a key role in transformation of B-lymphocytes mediated by Epstein-Barr virus (EBV) and can induce tumor formation in transgenic mice. However, the precise mechanism underlying EBNA2-mediated tumorigenesis remains elusive. Here, we report that EBNA2 can compromise mitotic spindle checkpoint (MSC) induced by the spindle inhibitor nocodazole and cause chromosomal instability (CIN) in HEp-2, U2-OS and BJAB cells. When EBNA2-expressing cells were treated with nocodazole, they exited mitosis prematurely and initiated another round of DNA synthesis. Nucleolocalization of EBNA2 was essential for EBNA2 to compromise MSC and to cause CIN. The metaphase chromosome spread data indicated that the EBNA2 expressing U2-OS cells showed a more heterogenous chromosome number distribution than the vector-transfected and parental cells. The median chromosome number for EBNA2-expressing, vector-transfected and parental U2-OS cells is 75, 65 and 64, respectively. EBNA2 was shown to be able to downregulate mitotic arrest deficient 2 (MAD2) approximately 2- to 3-fold and upregulate polo-like kinase 1 (PLK1) approximately 2-fold. The dysregulation of MAD2 and PLK1 may lead to activation of anaphase promoting complex/cyclosome and premature degradation of securin. Indeed, we found that when MSC was induced by nocodazole, securin was prematurely degraded in EBNA2-expressing cells. Finally, we show that EBNA2 could induce micronuclei and multinuclei formation in HEp-2 and U2-OS cells. Together, these studies reveal a new function of EBNA2 in cell-cycle regulation and may shed light on the role of EBNA2 in EBV-mediated tumorigenesis. PMID- 19126643 TI - CCDC62/ERAP75 functions as a coactivator to enhance estrogen receptor beta mediated transactivation and target gene expression in prostate cancer cells. AB - Human prostate cancer (PCa) and prostate epithelial cells predominantly express estrogen receptor (ER) beta, but not ERalpha. ERbeta might utilize various ER coregulators to mediate the E2-signaling pathway in PCa. Here, we identified coiled-coil domain containing 62 (CCDC62)/ERAP75 as a novel ER coactivator. CCDC62/ERAP75 is widely expressed in PCa cell lines and has low expression in MCF7 cells. Both in vitro and in vivo interaction assays using mammalian two hybrid, glutathione S-transferase pull-down and coimmunoprecipitation methods proved that ERbeta can interact with the C-terminus of CCDC62/ERAP75 via the ligand-binding domain. The first LXXLL motif within CCDC62/ERAP75 is required for the interaction between ERbeta and CCDC62/ERAP75. Electrophoretic mobility shift assay showed that CCDC62/ERAP75 can be recruited by the estrogen response element ER complex in the presence of ligand. Furthermore, a chromatin immunoprecipitation assay demonstrated the hormone-dependent recruitment of CCDC62/ERAP75 within the promoter of the estrogen-responsive gene cyclin D1. In addition, using silencing RNA (siRNA) against endogeneous CCDC62/ERAP75, we demonstrated that inhibition of endogenous CCDC62/ERAP75 results in the suppression of ERbeta-mediated transactivation as well as target gene expression in LNCaP cells. More importantly, using the tet-on overexpression system, we showed that induced expression of CCDC62/ERAP75 can enhance the E2-regulated cyclin D1 expression and cell growth in LNCaP cells. Together, our results revealed the role of CCDC62/ERAP75 as a novel coactivator in PCa cells that can modulate ERbeta transactivation and receptor function. PMID- 19126644 TI - A novel factor distinct from E2F mediates C-MYC promoter activation through its E2F element during exit from quiescence. AB - Although C-MYC is overexpressed in a number of tumors, the mechanisms governing its expression in normal or tumor cells are not completely understood. Recruitment of the Retinoblastoma protein family members to gene promoters by E2F factors has a dominant negative effect on their activity during the G(0) and G(1) phases of the cell cycle. Despite the presence of an E2F-binding site on the C MYC promoter, it escapes the repressive effect of E2F-Retinoblastoma complexes through unknown mechanisms during exit from quiescence. We hypothesized that occupancy of E2F elements by factors distinct from E2F might account for this escape. To test this hypothesis, we investigated whether the E2F element in the C MYC promoter is regulated differently than E2F elements in promoters that are activated at the G(1)-S transition. Employing gel shift analysis, the E2F element from the C-MYC promoter was found to form a unique non-E2F complex, referred to as E2F C-MYC Specific (EMYCS), which is not observed with E2F elements from several other promoters. The DNA contact residues for EMYCS are distinct but overlapping with residues required for binding of E2F proteins. Finally, the approximate estimated molecular weight of the DNA-binding component of EMCYS is 105 kDa. Functional studies indicate that EMYCS has transcriptional transactivation capacity and suggest that it is required to activate the C-MYC promoter during exit from quiescence. PMID- 19126645 TI - Differential roles for membrane-bound and soluble syndecan-1 (CD138) in breast cancer progression. AB - The heparan sulfate proteoglycan syndecan-1 (Sdc1) modulates cell proliferation, adhesion, migration and angiogenesis. Proteinase-mediated shedding converts Sdc1 from a membrane-bound coreceptor into a soluble effector capable of binding the same ligands. In breast carcinomas, Sdc1 overexpression correlates with poor prognosis and an aggressive phenotype. To distinguish between the roles of membrane-bound and shed forms of Sdc1 in breast cancer progression, human MCF-7 breast cancer cells were stably transfected with plasmids overexpressing wild type (WT), constitutively shed and uncleavable forms of Sdc1. Overexpression of WT Sdc1 increased cell proliferation, whereas overexpression of constitutively shed Sdc1 decreased proliferation. Fibroblast growth factor-2-mediated mitogen activated protein kinase signaling was reduced following small-interfering RNA (siRNA)-mediated knockdown of Sdc1 expression. Constitutively, membrane-bound Sdc1 inhibited invasiveness, whereas soluble Sdc1 promoted invasion of MCF-7 cells into matrigel matrices. The latter effect was reversed by the matrix metalloproteinase inhibitors N-isobutyl-N-(4-methoxyphenylsufonyl) glycyl hydroxamic acid and tissue inhibitor of metalloproteinase (TIMP)-1. Affymetrix microarray analysis identified TIMP-1, Furin and urokinase-type plasminogen activator receptor as genes differentially regulated in soluble Sdc1 overexpressing cells. Endogenous TIMP-1 expression was reduced in cells overexpressing soluble Sdc1 and increased in those overexpressing the constitutively membrane-bound Sdc1. Moreover, E-cadherin protein expression was downregulated in cells overexpressing soluble Sdc1. Our results suggest that the soluble and membrane-bound forms of Sdc1 play different roles at different stages of breast cancer progression. Proteolytic conversion of Sdc1 from a membrane bound into a soluble molecule marks a switch from a proliferative to an invasive phenotype, with implications for breast cancer diagnostics and potential glycosaminoglycan-based therapies. PMID- 19126647 TI - Overexpression of phospholipase D enhances matrix metalloproteinase-2 expression and glioma cell invasion via protein kinase C and protein kinase A/NF-kappaB/Sp1 mediated signaling pathways. AB - Glioblastoma is a severe type of primary brain tumor, and its highly invasive character is considered to be a major therapeutic obstacle. Phospholipase D (PLD) isozyme is overexpressed in various human tumor tissues and involved in tumorigenesis. However, the molecular mechanisms by which PLD enhances glioma invasion are unknown. In this study, we demonstrate that the increased expression of PLD and its enzymatic activity in the glioma stimulate the secretion and expression of matrix metalloproteinase (MMP)-2 and induce the invasiveness of glioma cells. The upregulation of MMP-2 induced by phosphatidic acid (PA), the product of PLD, was mediated by protein kinase C (PKC), protein kinase A (PKA), nuclear factor-kappaB (NF-kappaB) and Sp1 and it enhanced glioma cell invasion. PA activated PKC and PKA and induced the nuclear translocation and transactivation of NF-kappaB. PA also increased the binding of NF-kappaB and Sp1 to the MMP-2 promoter. Mutation of the NF-kappaB- or Sp1-binding sites significantly attenuated MMP-2 promoter activity. This is the first report to show that NF-kappaB and Sp1 are essential transcriptional factors linking PLD to MMP-2 upregulation, providing evidence that PLD contributes to glioma progression by enhancing MMP-2 expression and tumor cell invasion via PKC/PKA/NF-kappaB/Sp1 mediated signaling pathways. PMID- 19126646 TI - Genetic polymorphisms in the cytokine genes and risk of hepatocellular carcinoma in low-risk non-Asians of USA. AB - Polymorphisms in cytokine genes responsible for inflammatory and immune responses are associated with risk of hepatocellular carcinoma (HCC) in high-risk Chinese population. Similar data in low-risk populations are lacking. A population-based case-control study of HCC was conducted including 120 HCC patients and 230 matched control subjects of non-Asian residents in Los Angeles County, California. Genetic variants in the interferon gamma (IFNgamma), tumor necrosis factor-alpha (TNFalpha), interleukin-2 (IL-2), IL-4, IL-6, IL-10, IL-12 and IL-18 genes were determined by Taqman assays. The logistic regression method was used to analyze the data. For T helper (Th) 1 genes (IFNgamma, IL-6 and IL-12), relative to the putative high-activity genotypes, individual low-activity genotypes were associated with statistically non-significant increases in HCC risk. The odds ratio (OR) was 1.53 [95% confidence interval (CI) = 0.53-4.39] for three versus zero low-activity genotypes. For Th2 cytokines (IL-4 and IL-10), low versus high-activity genotypes were associated with statistically non significant decreases in HCC risk. The OR was 0.64 (95% CI = 0.27-1.55) for two versus zero low-activity genotypes. When the Th1 and Th2 genotypes were examined simultaneously, the highest level of risk was observed in individuals jointly possessing the highest number of low-activity Th1 genotypes and the lowest number of low-activity Th2 genotypes. There was a roughly doubling of risk between these two extreme genetic profiles, which did not reach statistical significance (OR = 1.98, 95% CI = 0.50-7.84, P = 0.08). In contrast to high-risk Chinese, Th1 and Th2 genotypes did not impact in a major way on risk of HCC in USA non-Asians. PMID- 19126648 TI - Cyr61 increases migration and MMP-13 expression via alphavbeta3 integrin, FAK, ERK and AP-1-dependent pathway in human chondrosarcoma cells. AB - Cysteine-rich 61 (Cyr61), from the CCN gene family, is a secreted and matrix associated protein, which is involved in many cellular activities such as growth and differentiation. However, the effect of Cyr61 on migration activity in human chondrosarcoma cells is mostly unknown. Here, we found that Cyr61 increased the migration and expression of matrix metalloproteinase (MMP)-13 in human chondrosarcoma cells (JJ012 cells). RGD peptide, alphavbeta3 monoclonal antibody and mitogen-activated protein kinase (MEK) inhibitors (PD98059 and U0126) but not RAD peptide inhibited the Cyr61-induced increase of the migration and MMP-13 upregulation of chondrosarcoma cells. Cyr61 stimulation increased the phosphorylation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK). In addition, activator protein-1 (AP-1) decoy oligodeoxynucleotide also suppressed the MMP-13 messenger RNA and enzyme activity enhanced by Cyr61. Moreover, Cyr61 increased the binding of c-Fos and c-Jun to the AP-1 element on the MMP-13 promoter. Taken together, our results indicated that Cyr61 enhances the migration of chondrosarcoma cells by increasing MMP-13 expression through the alphavbeta3 integrin receptor, FAK, ERK, c-Fos/c-Jun and AP-1 signal transduction pathway. PMID- 19126649 TI - Elevated NCOR1 disrupts a network of dietary-sensing nuclear receptors in bladder cancer cells. AB - Increasingly invasive bladder cancer cells lines displayed insensitivity toward a panel of dietary-derived ligands for members of the nuclear receptor superfamily. Insensitivity was defined through altered gene regulatory actions and cell proliferation and reflected both reduced receptor expression and elevated nuclear receptor corepressor 1 (NCOR1) expression. Stable overexpression of NCOR1 in sensitive cells (RT4) resulted in a panel of clones that recapitulated the resistant phenotype in terms of gene regulatory actions and proliferative responses toward ligand. Similarly, silencing RNA approaches to NCOR1 in resistant cells (EJ28) enhanced ligand gene regulatory and proliferation responses, including those mediated by peroxisome proliferator-activated receptor (PPAR) gamma and vitamin D receptor (VDR) receptors. Elevated NCOR1 levels generate an epigenetic lesion to target in resistant cells using the histone deacetylase inhibitor vorinostat, in combination with nuclear receptor ligands. Such treatments revealed strong-additive interactions toward the PPARgamma, VDR and Farnesoid X-activated receptors. Genome-wide microarray and microfluidic quantitative real-time, reverse transcription-polymerase chain reaction approaches, following the targeting of NCOR1 activity and expression, revealed the selective capacity of this corepressor to govern common transcriptional events of underlying networks. Combined these findings suggest that NCOR1 is a selective regulator of nuclear receptors, notably PPARgamma and VDR, and contributes to their loss of sensitivity. Combinations of epigenetic therapies that target NCOR1 may prove effective, even when receptor expression is reduced. PMID- 19126650 TI - Oncogenic transformation of human ovarian surface epithelial cells with defined cellular oncogenes. AB - Ovarian surface epithelium (OSE) is considered to give rise to epithelial ovarian carcinomas (EOCs). To elucidate early processes contributing to the development of EOCs from the OSE, two batches of primary human OSE cells were transduced with non-viral human genes (mutant Cdk4, cyclinD1 and hTERT) so as to efficiently establish normal diploid OSE cells without chromosomal instability. Then defined genetic alterations frequently observed in EOCs were transduced into the OSE cells. A combination of p53 inactivation and oncogenic Kras transduction did not confer tumor-forming ability in immunodeficient mice, though additional transduction of Akt or combined transduction of c-myc with bcl-2 did result in tumor formation. In the latter case, tumors demonstrated phenotypes reminiscent of human EOCs, including cytokeratin expression, a highly aggressive phenotype, metastatic behavior and formation of ascites. These results indicate that inactivation of p53 and activation of the Ras pathway play critical roles in ovarian carcinogenesis in co-operation with the Akt or c-myc pathways. This first in vitro model system faithfully recapitulating the development of EOCs using normal human OSE cells should greatly facilitate further studies of EOCs. PMID- 19126652 TI - Epigenetic profiling reveals etiologically distinct patterns of DNA methylation in head and neck squamous cell carcinoma. AB - Head and neck squamous cell carcinomas (HNSCCs) represent clinically and etiologically heterogeneous tumors affecting >40 000 patients per year in the USA. Previous research has identified individual epigenetic alterations and, in some cases, the relationship of these alterations with carcinogen exposure or patient outcomes, suggesting that specific exposures give rise to specific types of molecular alterations in HNSCCs. Here, we describe how different etiologic factors are reflected in the molecular character and clinical outcome of these tumors. In a case series of primary, incident HNSCC (n = 68), we examined the DNA methylation profile of 1413 autosomal CpG loci in 773 genes, in relation to exposures and etiologic factors. The overall pattern of epigenetic alteration could significantly distinguish tumor from normal head and neck epithelial tissues (P < 0.0001) more effectively than specific gene methylation events. Among tumors, there were significant associations between specific DNA methylation profile classes and tobacco smoking and alcohol exposures. Although there was a significant association between methylation profile and tumor stage (P < 0.01), we did not observe an association between these profiles and overall patient survival after adjustment for stage; although methylation of a number of specific loci falling in different cellular pathways was associated with overall patient survival. We found that the etiologic heterogeneity of HNSCC is reflected in specific patterns of molecular epigenetic alterations within the tumors and that the DNA methylation profiles may hold clinical promise worthy of further study. PMID- 19126653 TI - SNPs in ultraconserved elements and familial breast cancer risk. PMID- 19126654 TI - The DNA repair protein NBS1 influences the base excision repair pathway. AB - NBS1 fulfills important functions for the maintenance of genomic stability and cellular survival. Mutations in the NBS1 (Nijmegen Breakage Syndrome 1) gene are responsible for the Nijmegen breakage syndrome (NBS) in humans. The symptoms of this disease and the phenotypes of NBS1-defective cells, especially their enhanced radiosensitivity, can be explained by an impaired DNA double-strand break-induced signaling and a disturbed repair of these DNA lesions. We now provide evidence that NBS1 is also important for cellular survival after oxidative or alkylating stress where it is required for the proper initiation of base excision repair (BER). NBS1 downregulated cells show reduced activation of poly-(adenosine diphosphate-ribose)-polymerase-1 (PARP1) following genotoxic treatment with H(2)O(2) or methyl methanesulfonate, indicating impaired processing of damaged bases by BER as PARP1 activity is stimulated by the single strand breaks intermediately generated during this repair pathway. Furthermore, extracts of these cells have a decreased capacity for the in vitro repair of a double-stranded oligonucleotide containing either uracil or 8-oxo-7,8 dihydroguanine to trigger BER. Our data presented here highlight for the first time a functional role for NBS1 in DNA maintenance by the BER pathway. PMID- 19126655 TI - Death receptor pathways mediate targeted and non-targeted effects of ionizing radiations in breast cancer cells. AB - Delayed cell death by mitotic catastrophe is a frequent mode of solid tumor cell death after gamma-irradiation, a widely used treatment of cancer. Whereas the mechanisms that underlie the early gamma-irradiation-induced cell death are well documented, those that drive the delayed cell death are largely unknown. Here we show that the Fas, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and tumor necrosis factor (TNF)-alpha death receptor pathways mediate the delayed cell death observed after gamma-irradiation of breast cancer cells. Early after irradiation, we observe the increased expression of Fas, TRAIL-R and TNF-R that first sensitizes cells to apoptosis. Later, the increased expression of FasL, TRAIL and TNF-alpha permit the apoptosis engagement linked to mitotic catastrophe. Treatments with TNF-alpha, TRAIL or anti-Fas antibody, early after radiation exposure, induce apoptosis, whereas the neutralization of the three death receptors pathways impairs the delayed cell death. We also show for the first time that irradiated breast cancer cells excrete soluble forms of the three ligands that can induce the death of sensitive bystander cells. Overall, these results define the molecular basis of the delayed cell death of irradiated cancer cells and identify the death receptors pathways as crucial actors in apoptosis induced by targeted as well as non-targeted effects of ionizing radiation. PMID- 19126656 TI - Super competition as a possible mechanism to pioneer precancerous fields. AB - Cancer is the result of sequential genetic changes over time that transform a cell into a malignant and ultimately invasive entity. The insight that cancerous cells arise from a series of mutations in oncogenes and tumor suppressors, commonly known as multistep carcinogenesis, has been conceptually elaborated and proven in the last 20 years. Although knowledge about late steps of cancerogenesis and disease progression has greatly advanced, the initial molecular events remain largely unknown. Basic research in Drosophila has started the quest to find early markers that detect initial clonal expansion of precancerous cells. These efforts were spurred by novel findings demonstrating that certain mutations transform cells into super-competitors that expand at the expense of the surrounding epithelial cells without inducing histological changes. This mechanism, discovered as super competition in the fly, might also lie at the heart of a clinical observation termed 'field cancerization'. This review aims to bring together current understanding from basic research on cell competition and clinical studies that have analyzed field characteristics to highlight parallels and possible connections. PMID- 19126657 TI - Coalescence of splines on a basket mapping catheter during ablation using a closed-loop irrigation catheter. PMID- 19126658 TI - Experimental models for nephropathy. AB - Nephropathy is a leading cause of morbidity and mortality and its prevalence is continuously increasing in industrialised nations. Nephropathy is characterised to varying degrees by nodular glomerulosclerosis, glomerular basement membrane thickness and mesangial expansion, leading to a decline in glomerular filtration rate, persistent elevated albuminuria, elevated arterial blood pressure and fluid retention. Hyperglycaemia, hyperlipidaemia and hypertension are considered to be the major risk factors implicated in the progression of nephropathy. Various signalling systems, such as vasoconstrictor peptides, inflammatory mediators, growth factors and adhesion molecules, are involved in the pathogenesis of nephropathy. At present, no promising therapy is available to treat patients with nephropathy due to lack of understanding of signalling culprits involved in the pathogenesis of nephropathy. Animal models are being developed to better understand the disease pathogenesis and develop drugs for nephropathy. In the present review, we have discussed various animal models for nephropathy, which may open vistas for developing new drugs to treat nephropathy. PMID- 19126659 TI - Up- and down-regulation of angiotensin II AT1-A and AT1-B receptors in afferent and efferent rat kidney arterioles. AB - INTRODUCTION: The contractile effect of angiotensin II via AT1 receptors on the kidney arterioles is a crucial element in the kidney microcirculation. Angiotensin II also plays a role as an inhibitor via the AT1 receptors in the renin granulation of the arterioles. We have previously demonstrated that the AT1 receptors are downregulated in the renin-positive smooth muscle cells (SMCs) in contrast to renin-negative SMCs. In this study, we estimated the numbers of the AT1 receptor sub-types separately in the afferent and efferent arterioles and the renin-positive and renin-negative SMCs. METHODS: The immunohistochemical signals of the AT1-A and AT1-B receptors were counted by stereological methods. (1) RESULTS: The number of AT1-B receptors in the efferent arterioles (expressed in signals/microm3; mean (CV): 0.32 (0.33)) was significantly higher (78%; p<0.05) as compared with the number in the afferent arterioles (0.18 (0.11)). No differences were found in the AT1-A receptors. In a number of AT1-A receptors, significant differences (p<0.01) were detected between the afferent arteriolar renin-positive SMCs (0.13 (0.36)) and the number in renin-negative SMCs (0.25 (0.34)). The AT1-B receptors did not display any differences. CONCLUSIONS: These results indicate that the AT1 receptor sub-types are regulated independently in the SMCs of the normal kidney arterioles. PMID- 19126660 TI - Association of renin-angiotensin system and natriuretic peptide receptor A gene polymorphisms with hypertension in a Hellenic population. AB - INTRODUCTION: Hypertension results from the interaction of genetic and environmental factors. Since the renin-angiotensin and the natriuretic peptide systems contribute to blood pressure regulation, variations in the relative genes are candidates for the development of hypertension. MATERIALS AND METHODS: In 194 hypertensives and 304 controls of Hellenic origin, the possible association between the (CA)n repeat polymorphism of angiotensinogen (AGT), the 250 bp insertion/deletion (I/D) of angiotensin-converting enzyme (ACE), the tetranucleotide repeat polymorphism (TCTG)n of renin, and the (CT)n repeat polymorphism of the natriuretic peptide receptor A (NPRA) and hypertension was assessed. RESULTS: No association between AGT and NPRA polymorphisms and hypertension was observed. The presence of ID or DD genotype of ACE was associated with an increased risk for hypertension compared with the II genotype (OR: 1.782 [95% CI: 1.032-3.077]), whereas the LL genotype of the renin gene was associated with a decreased risk compared with the SS genotype (OR: 0.174 [95% CI: 0.044-0.689]). However, after adjustment for confounding factors only the latter association remained. CONCLUSIONS: In the present study conducted in a homogeneous Hellenic population, no associations between AGT,ACE, and NPRA gene polymorphisms and hypertension were found. The presence of a significant negative association between the LL polymorphism of the renin gene and hypertension requires further confirmation. PMID- 19126661 TI - Association of insertion/deletion polymorphism of angiotensin-converting enzyme gene with essential hypertension and type 2 diabetes mellitus in Malaysian subjects. AB - INTRODUCTION: The deletion (D) allele of the angiotensin-converting enzyme (ACE) gene has been studied in various populations in relation to hypertension (HTN) and type 2 diabetes mellitus (T2DM) with contradictory results. This study sought to determine the association of insertion (I)/D polymorphism of the ACE gene in hypertensive and T2DM subjects in a Malaysian population. MATERIALS AND METHODS: A total of 260 subjects consisting of 65 HTN, 60 T2DM, 65 T2DM with HTN and 70 controls were recruited. Genotyping was performed by polymerase chain reaction initially and mistyping of DD genotypes was performed with an insertion-specific primer. RESULTS: The frequency for II, ID and DD genotypes of the ACE gene was 36.92%, 52.31% and 10.77% in HTN, 40.00%, 41.67% and 18.33% inT2DM, 30.77%, 53.85% and 15.38% inT2DM with HTN and 57.14%, 40.00% and 2.86% in controls, respectively. The frequency for the D allele was 36.92% in HTN, 39.17% in T2DM and 42.31% in T2DM with HTN compared to 22.86% in controls. The genotype and allele frequency of the ACE gene polymorphism differed significantly in patients when compared to controls (p < 0.05). CONCLUSION: The D allele of the ACE gene is associated with essential HTN and T2DM in Malaysian subjects. PMID- 19126662 TI - Angiotensin-converting enzyme gene polymorphism in arrhythmogenic right ventricular dysplasia: is DD genotype helpful in predicting syncope risk? AB - INTRODUCTION: Arrhythmogenic right ventricular dysplasia (ARVD) is a heritable disorder characterised by fibrofatty replacement of right ventricular myocytes and increased risk of ventricular arrhythmias and sudden cardiac death. Angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism affects myocardial ACE levels. DD genotype favours myocardial fibrosis and is associated with malignant ventricular tachycardia. The aim of this study was to explore ACE gene polymorphism in ARVD patients. METHODS: Twenty-nine patients with ARVD and 24 controls were included. All ARVD patients had documented sustained ventricular tachycardia. Thirteen patients had syncopal episodes. Six patients were resuscitated from sudden cardiac death. ACE gene polymorphism was identified by polymerase chain reaction technique. RESULTS: There was no significant difference in DD genotype frequency between ARVD patients and controls (44.8% vs. 45.8%, p=0.94). However, DD genotype frequency was significantly higher in ARVD patients with syncopal episodes compared to those without syncope (69.2% vs. 25.0%, p=0.017, odds ratio:6.750, 95% confidence interval: 1.318-34.565). DD genotype was detected in higher frequency also in patients with a family history of sudden cardiac death (66.7% vs. 39.1%,p=0.36). CONCLUSION: High prevalence of DD genotype in ARVD patients with syncope suggests that ACE I/D polymorphism might be useful in identifying high-risk patients for syncope. PMID- 19126663 TI - The bradykinin-degrading aminopeptidase P is increased in women taking the oral contraceptive pill. AB - INTRODUCTION: The renin-angiotensin and kininogen-kinin hormonal systems are critically involved in regulating blood pressure and are candidates in contributing to oral contraceptive pill (OCP)-induced hypertension. Angiotensin converting enzyme (ACE) and aminopeptidase P (AP-P) are key enzymes in these systems and are both involved in the degradation of the vasodilator bradykinin. METHODS: Circulating ACE and AP-P levels were measured by activity assay using selective fluorogenic peptide substrates in plasma samples from the Leeds Family Study. In addition, the effect of progesterone on the expression of AP-P and ACE was examined in cells. RESULTS: Women on the OCP had higher age-adjusted plasma AP-P (mean [95% confidence interval]) (0.27 [0.23-0.32] nmol/min/ml (n = 53)) compared with women not on the OCP (0.17 [0.16-0.19] nmol/min/ml (n = 133), p < 0.001) or males (0.19 [0.17-0.20] nmol/min/ml (n = 209), p<0.001). There were no differences in the age-adjusted plasma ACE levels among the three groups. In HepG2 cells, progesterone treatment increased the AP-P protein and mRNA expression, whereas no effect of progesterone treatment was observed for ACE. CONCLUSION: Increased AP-P may result in increased breakdown of bradykinin. These data suggest that progesterone-induced increases in AP-P may contribute to the development of OCP-induced hypertension in susceptible Women. PMID- 19126664 TI - The angiotensin AT4 receptor subtype as a target for the treatment of memory dysfunction associated with Alzheimer's disease. AB - Over recent years antihypertensive drugs, particularly angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs), have been reported to have beneficial effects upon cognitive impairment. Such findings suggest that pharmacological manipulation of angiotensin ligands may be of clinical importance in slowing or halting the cognitive deterioration seen in vascular dementia and Alzheimer's disease. The mechanism(s) underlying these improvements in cognitive function remains unclear; however, important leads are emerging. The angiotensin AT4 receptor subtype, discovered by our laboratory in 1992, influences several important behaviours and physiologies, including learning and memory, and may play a role in this cognitive improvement. This review initially describes the therapeutic drugs approved by the Federal Drug Administration and new approaches presently being developed to treat Alzheimer's disease-induced cognitive impairment. Next, the biologically-active angiotensin ligands and their respective receptor subtypes are discussed, followed by the roles of angiotensin II, angiotensin IV, ACE inhibitors and ARBs in cognitive function. We conclude with a working hypothesis concerning the importance of the AT4 receptor subtype as a new potential drug target for the treatment of Alzheimer's disease-associated memory loss. PMID- 19126665 TI - Chronic angiotensin-converting enzyme inhibition up-regulates mouse kidney growth arrest specific-6 protein and the AXL subfamily of receptor tyrosine kinases. AB - INTRODUCTION: Growth arrest specific-6 (GAS-6), a vitamin K-dependent protein, is a potential mediator in progressive and chronic renal disease, specifically as a mediator of abnormal mesangial cell proliferation. Nitric oxide and angiotensin II affect mesangial cell proliferation. However, an association between nitric oxide synthase or angiotensin II on GAS-6 expression in the kidney has not previously been examined. Thus, our aim was to examine the effects of antihypertensive angiotensin-converting enzyme inhibitors and chronic nitric oxide synthase inhibition on the kidney expression of GAS-6 and its receptors AXL, MER and RSE. METHODS: Four groups of adult male C57BL/6J mice were studied: group 1, untreated controls (tap water for six weeks); group 2, treated orally with a nitric oxide synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME, 0.325 mg/ml for six weeks); group 3, treated orally with captopril (0.6875 mg/ml for six weeks); group 4, co-treated orally with L-NAME and captopril (same doses for six weeks). At the end of the study, kidneys were placed in fixative and processed to paraffin for immunohistochemical staining. RESULTS: GAS-6 and its receptors were not present in control and L-NAME-treated mice. Positive GAS-6 staining was detectable only in those mice receiving some form of chronic dosing with captopril, whether they were treated with captopril only or with captopril and L-NAME. Immunohistochemical detection across cases for MER and RSE was rare, whereas AXL-positive staining in the kidney mirrored GAS-6 staining/expression. The staining of GAS6 and AXL was predominantly localised to the renal tubular cells. CONCLUSIONS: These findings suggest that GAS-6 may not be a final common pathway for nitric oxide synthase inhibition-induced renal disease. Renal tubular GAS-6 expression following captopril treatment was unexpected and could be beneficial in preventing tubular atrophy following the onset of persistent systemic hypertension. PMID- 19126666 TI - Renin-angiotensin system-related highlights from the High Blood Pressure Research Conference annual meeting. AB - The High Blood Pressure Research Conference of the American Heart Association is a theoretical meeting for hypertension researchers who direct their attention to hypertension-related basic disease mechanisms. The items that I have selected for this brief review are molecular intracellular receptor function, novel angiotensin (Ang)-related pathways, including Ang-(1-7), the Mas receptor, and angiotensin-converting enzyme 2, oxidative stress, immunity, the (pro)renin receptor, and until now unappreciated signalling pathways, such as the tonicity element binding protein. PMID- 19126667 TI - Questions & answers--understanding the renin system. PMID- 19126668 TI - Failure of linezolid therapy for post-neurosurgical meningitis due to Enterococcus faecium. PMID- 19126669 TI - Analysis of 4758 Escherichia coli bacteraemia episodes: predictive factors for isolation of an antibiotic-resistant strain and their impact on the outcome. AB - OBJECTIVES: To describe the predictive factors for the isolation of fluoroquinolone-resistant or extended- spectrum beta-lactamase (ESBL)-producing Escherichia coli and their impact on bacteraemia outcome. METHODS: Analysis of E. coli bacteraemia episodes prospectively collected through a blood culture surveillance programme from January 1991 to December 2007. RESULTS: Out of 18 080 episodes, 4758 (26%) E. coli bacteraemias were reported in the period of study. Mortality was noted in 440 cases (9%). Fluoroquinolone-resistant strains were reported in 1300 (27%) cases and ESBL-producing strains in 211 cases (4%). One hundred and seventy-eight strains out of 211 (84%) ESBL-producing E. coli were isolated since 2001. The two main independent risk factors for mortality were shock (OR: 10.28, P < 0.001) and inappropriate empirical therapy (OR: 4.83, P < 0.001). Inappropriate empirical therapy was significantly more frequent for infections caused by fluoroquinolone-resistant strains (n = 203, 16%, P < 0.001) and ESBL-producing strains (n = 110, 52%, P < 0.001). Independent factors associated with the isolation of a fluoroquinolone-resistant strain were: nosocomial origin (OR: 1.61, P < 0.001); urinary catheterization (OR: 2.44, P < 0.001); and previous therapy with a fluoroquinolone (OR: 7.41, P < 0.001). The independent risk factors associated with the isolation of an ESBL-producing strain were: nosocomial origin (OR: 1.68, P = 0.03); urinary catheterization (OR: 1.88, P = 0.001); and previous beta-lactam antibiotic therapy (OR: 2.81, P < 0.001). CONCLUSIONS: Inappropriate empirical therapy was the strongest independent factor that we could modify to improve mortality in E. coli bacteraemia and was more frequent in cases caused by fluoroquinolone-resistant or ESBL-producing strains. Nosocomial acquisition, urinary catheterization and previous therapy with a fluoroquinolone or beta-lactam were predictive factors for infection with an antibiotic-resistant strain. PMID- 19126670 TI - Cinnamon extract protects against acute alcohol-induced liver steatosis in mice. AB - Acute and chronic consumption of alcohol can cause increased intestinal permeability and bacterial overgrowth, thereby increasing portal endotoxin levels. This barrier impairment subsequently leads to an activation of hepatic Kupffer cells and increased release of reactive oxygen species as well as of tumor necrosis factor-alpha (TNFalpha). Recent studies have suggested that cinnamon extract may have antiinflammatory effects. In the present study, the protective effects of an alcoholic extract of cinnamon bark was assessed in a mouse model of acute alcohol-induced steatosis and in RAW 264.7 macrophages, used here as a model of Kupffer cells. Acute alcohol ingestion caused a >20-fold increase in hepatic lipid accumulation. Pretreatment with cinnamon extract significantly reduced the hepatic lipid accumulation. This protective effect of cinnamon extract was associated with an inhibition of the induction of the myeloid differentiation primary response gene (MyD) 88, inducible nitric oxide (NO) synthase (iNOS), and plasminogen activator inhibitor 1 mRNA expression found in livers of alcohol-treated animals. In vitro prechallenge with cinnamon extract suppressed lipopolysaccharide (LPS)-induced MyD88, iNOS, and TNFalpha expression as well as NO formation almost completely. Furthermore, LPS treatment of RAW 264.7 macrophages further resulted in degradation of inhibitor kappaB; this effect was almost completely blocked by cinnamon extract. Taken together, our data show that an alcohol extract of cinnamon bark may protect the liver from acute alcohol-induced steatosis through mechanisms involving the inhibition of MyD88 expression. PMID- 19126671 TI - Partial replacement of dietary (n-6) fatty acids with medium-chain triglycerides decreases the incidence of spontaneous colitis in interleukin-10-deficient mice. AB - Enteral nutrition has a primary therapeutic effect in active Crohn's disease. It is unknown which nutrient(s) account for this action, but a role for both the amount and type of dietary fat has been postulated. Some clinical and experimental data suggest that medium-chain triglycerides (MCT) may reduce intestinal inflammation. We aimed to assess the effect of replacing part of the dietary fat with MCT on the incidence and severity of colitis in interleukin (IL) 10(-/-) mice under specific pathogen-free conditions. Twenty-four IL-10(-/-) 4-wk old mice were randomized to receive a control diet based on sunflower oil [(n-6) fatty acids (FA)] and an experimental isocaloric, isonitrogenous diet with 50% sunflower and 50% coconut oil (MCT diet). When the mice were 12 wk old, they were killed and the colon was examined for the presence of colitis, lymphocyte subpopulations and apoptosis, ex vivo cytokine production in supernatant of colon explants, toll-like receptor (TLR)-2 and TLR-9 mRNA, and FA profile in colonic tissue homogenates. Colitis incidence was lower in the IL-10(-/-) mice fed the MCT diet (1/12) than in the mice fed the control diet (8/12; P = 0.03). The histological damage score was also lower in the former (P < 0.0005). Feeding the MCT diet resulted in fewer total and apoptotic intraepithelial CD3+ and lamina propria CD3+CD4+ lymphocytes, as well as downregulated production of IL-6 and interferon-gamma, and reduced TLR-9 mRNA. We conclude that partial replacement of dietary (n-6) FA with MCT decreases the incidence of colitis in a model of spontaneous intestinal inflammation and provide experimental arguments for a possible primary therapeutic effect of MCT in human Crohn's disease. PMID- 19126672 TI - Spontaneous phosphoinositide 3-kinase signaling dynamics drive spreading and random migration of fibroblasts. AB - During directed cell migration (chemotaxis), cytoskeletal dynamics are stimulated and spatially biased by phosphoinositide 3-kinase (PI3K) and other signal transduction pathways. Live-cell imaging using total internal reflection fluorescence (TIRF) microscopy revealed that, in the absence of soluble cues, 3' phosphoinositides are enriched in a localized and dynamic fashion during active spreading and random migration of mouse fibroblasts on adhesive surfaces. Surprisingly, we found that PI3K activation is uncoupled from classical integrin mediated pathways and feedback from the actin cytoskeleton. Inhibiting PI3K significantly impairs cell motility, both in the context of normal spreading and when microtubules are dissociated, which induces a dynamic protrusion phenotype as seen by TIRF in our cells. Accordingly, during random migration, 3' phosphoinositides are frequently localized to regions of membrane protrusion and correlate quantitatively with the direction and persistence of cell movement. These results underscore the importance of localized PI3K signaling not only in chemotaxis but also in basal motility/migration of fibroblasts. PMID- 19126673 TI - Regulation of Rap1 activity is required for differential adhesion, cell-type patterning and morphogenesis in Dictyostelium. AB - Regulated cell adhesion and motility have important roles during growth, development and tissue homeostasis. Consequently, great efforts have been made to identify genes that control these processes. One candidate is Rap1, as it has been implicated in the regulation of adhesion and motility in cell culture. To further study the role of Rap1 during multicellular development, we generated a mutant in a potential Rap1 GTPase activating protein (RapGAPB) in Dictyostelium. rapGAPB(-) cells have increased levels of active Rap1 compared with wild-type cells, indicating that RapGAPB regulates Rap1 activity. Furthermore, rapGAPB(-) cells exhibit hallmark phenotypes of other known mutants with hyperactivated Rap1, including increased substrate adhesion and abnormal F-actin distribution. However, unlike these other mutants, rapGAPB(-) cells do not exhibit impaired motility or chemotaxis, indicating that RapGAPB might only regulate specific roles of Rap1. Importantly, we also found that RapGAPB regulates Rap1 activity during multicellular development and is required for normal morphogenesis. First, streams of aggregating rapGAPB(-) cells break up as a result of decreased cell cell adhesion. Second, rapGAPB(-) cells exhibit cell-autonomous defects in prestalk cell patterning. Using cell-type-specific markers, we demonstrate that RapGAPB is required for the correct sorting behaviour of different cell types. Finally, we show that inactivation of RapGAPB affects prestalk and prespore cell adhesion. We therefore propose that a possible mechanism for RapGAPB-regulated cell sorting is through differential adhesion. PMID- 19126674 TI - Combinatorial probabilistic chromatin interactions produce transcriptional heterogeneity. AB - Gene regulation often appears deterministic in the average cell population, but transcription is a probabilistic process at the single-cell level. Although many mechanisms are invoked to account for this behavior, it is difficult to determine how cell-to-cell variation in the interactions of transcription factors with target chromatin impact transcriptional output. Here, we use cells that contain a 200-copy tandem array of promoter or reporter gene units to simultaneously visualize transient interaction, equilibrium or steady-state binding of fluorescent-protein-labeled glucocorticoid receptor with its DNA response elements, the recruitment of diverse coregulators, and transcriptional output at the single-cell level. These regulatory proteins associate with target chromatin via a probabilistic mechanism that produces cell-to-cell variability in binding. The multiple steps of this process are partially independent and differ between individual regulators. The association level of each regulator influences the transcriptional output in individual cells, but this does not account for all transcriptional heterogeneity. Additionally, specific combinatorial interactions of the glucocorticoid receptor and coregulators with response elements regulate transcription at the single-cell level. Like many endogenous genes, the average array transcriptional activity evolves over time. This apparently deterministic average temporal promoter progression involves changes in the probability that specific combinatorial glucocorticoid receptor and coregulator interactions will occur on the response elements in single cells. These data support the emerging ;return-to-template' transcription model, which mechanistically unifies the observed extremely transient interactions between the transcription factor and response elements, cell-to-cell variability in steady-state association of factors with chromatin, and the resulting heterogeneous gene expression between individual cells. PMID- 19126675 TI - Connexin mutation that causes dominant congenital cataracts inhibits gap junctions, but not hemichannels, in a dominant negative manner. AB - The connexin (Cx) 50, E48K, mutation is associated with a human dominant congenital cataract; however, the underlying molecular mechanism has not been characterized. The glutamate (E) residue at position 48 is highly conserved across animal species and types of connexins. When expressed in paired Xenopus oocytes, human (h) and chicken (ch) Cx50 E48K mutants showed no electrical coupling. In addition, this mutation acts in a dominant negative manner when paired hetero-typically or hetero-merically with wild-type Cx50, but has no such effect on Cx46, the other lens fiber connexin. A similar loss-of-function and dominant negative effect was observed using dye transfer assays in the same system. By using two different dye transfer methods, with two different tracer dyes, we found chCx50 E48K expressed in chicken lens embryonic fibroblast cells by retroviral infection similarly failed to induce dye coupling, and prevented wild-type chCx50 from forming functional gap junctions. In contrast to its effect on gap junctions, the E48K mutation has no effect on hemichannel activity when assayed using electrical conductance in oocytes, and mechanically induced dye uptake in cells. Cx50 is functionally involved in cell differentiation and lens development, and the E48K mutant promotes primary lens cell differentiation indistinguishable from wild-type chCx50, despite its lack of junctional channel function. Together the data show that mutations affecting gap junctions but not hemichannel function of Cx50 can lead to dominant congenital cataracts in humans. This clearly supports the model of intercellular coupling of fiber cells creating a microcirculation of nutrients and metabolites required for lens transparency. PMID- 19126676 TI - Subcellular localization and dimerization of APLP1 are strikingly different from APP and APLP2. AB - The molecular association between APP and its mammalian homologs has hardly been explored. In systematically addressing this issue, we show by live cell imaging that APLP1 mainly localizes to the cell surface, whereas APP and APLP2 are mostly found in intracellular compartments. Homo- and heterotypic cis interactions of APP family members could be detected by FRET and co-immunoprecipitation analysis and occur in a modular mode. Only APLP1 formed trans interactions, supporting the argument for a putative specific role of APLP1 in cell adhesion. Deletion mutants of APP family members revealed two highly conserved regions as important for the protein crosstalk. In particular, the N-terminal half of the ectodomain was crucial for APP and APLP2 interactions. By contrast, multimerization of APLP1 was only partially dependent on this domain but strongly on the C-terminal half of the ectodomain. We further observed that coexpression of APP with APLP1 or APLP2 leads to diminished generation of Abeta42. The current data suggest that this is due to the formation of heteromeric complexes, opening the way for novel therapeutic strategies targeting these complexes. PMID- 19126677 TI - FAK engages multiple pathways to maintain survival of fibroblasts and epithelia: differential roles for paxillin and p130Cas. AB - Different cell types interpret their distinct extracellular matrix (ECM) environments to bring about specific cell fate decisions, and can differentiate or undergo apoptosis depending on their local adhesive interactions. Apoptosis in response to an inappropriate ECM environment is termed ;anoikis', or homelessness. Several studies, utilising a variety of cell types, have indicated a common, crucial role for focal adhesion kinase (FAK) in suppressing anoikis. A wide range of different integrins can activate FAK, raising the question of how cell type specific effects are regulated. In this study, we have used a constitutively active form of FAK to examine the mechanism of FAK-mediated survival signalling in cell types from distinct embryonic lineages that show differing sensitivities to anoikis. We demonstrate that both fibroblasts and epithelial cells prevent anoikis through FAK activation. We show that FAK activates multiple downstream pathways in order to suppress anoikis. However FAK regulates survival through a more restricted set of pathways in the more anoikis sensitive epithelial cells. Furthermore, we identify a novel role for paxillin in apoptosis suppression. PMID- 19126679 TI - Comment on 'cancer information for management'. PMID- 19126678 TI - Dynamic complexes of A-type lamins and emerin influence adipogenic capacity of the cell via nucleocytoplasmic distribution of beta-catenin. AB - It is well documented that adipogenic differentiation of the cell is associated with downregulation of Wnt/beta-catenin signalling. Using preadipocytes and dermal fibroblasts, we have found that activation of the adipogenic program was associated with marked changes in the expression of nuclear beta-catenin interacting partners, emerin and lamins A/C, to influence expression and activation of peroxisome proliferators-activated receptors gamma (PPARgamma). In addition, silencing of protein expression with siRNA revealed that beta-catenin and emerin influenced each other's levels of expression and the onset of adipogenesis, suggesting that changes in the expression of nuclear lamina proteins were intimately linked to the stability of beta-catenin. By contrast, dermal fibroblasts, which are emerin null, demonstrated increased nuclear accumulation of stable beta-catenin and constant lamin expression. This was also associated with an unusual adipogenic capacity of the cells, with adipogenesis occurring in the presence of activated beta-catenin but declining upon silencing of the protein expression with siRNA. We propose that the process of adipogenesis is affected by a dynamic link between complexes of emerin and lamins A/C at the nuclear envelope and nucleocytoplasmic distribution of beta-catenin, to influence cellular plasticity and differentiation. PMID- 19126680 TI - Microtubule plus-end tracking by CLIP-170 requires EB1. AB - Microtubules are polarized polymers that exhibit dynamic instability, with alternating phases of elongation and shortening, particularly at the more dynamic plus-end. Microtubule plus-end tracking proteins (+TIPs) localize to and track with growing microtubule plus-ends in the cell. +TIPs regulate microtubule dynamics and mediate interactions with other cellular components. The molecular mechanisms responsible for the +TIP tracking activity are not well understood, however. We reconstituted the +TIP tracking of mammalian proteins EB1 and CLIP 170 in vitro at single-molecule resolution using time-lapse total internal reflection fluorescence microscopy. We found that EB1 is capable of dynamically tracking growing microtubule plus-ends. Our single-molecule studies demonstrate that EB1 exchanges rapidly at microtubule plus-ends with a dwell time of <1 s, indicating that single EB1 molecules go through multiple rounds of binding and dissociation during microtubule polymerization. CLIP-170 exhibits lattice diffusion and fails to selectively track microtubule ends in the absence of EB1; the addition of EB1 is both necessary and sufficient to mediate plus-end tracking by CLIP-170. Single-molecule analysis of the CLIP-170-EB1 complex also indicates a short dwell time at growing plus-ends, an observation inconsistent with the copolymerization of this complex with tubulin for plus-end-specific localization. GTP hydrolysis is required for +TIP tracking, because end-specificity is lost when tubulin is polymerized in the presence of guanosine 5'-[alpha,beta methylene]triphosphate (GMPCPP). Together, our data provide insight into the mechanisms driving plus-end tracking by mammalian +TIPs and suggest that EB1 specifically recognizes the distinct lattice structure at the growing microtubule end. PMID- 19126681 TI - Consumers indirectly increase infection risk in grassland food webs. AB - Most pathogens exist within complicated food webs of interacting hosts, vectors, competitors, and predators. Although theory has demonstrated a variety of mechanisms by which predation and competition in food webs can indirectly control infection risk in hosts, there have until now been no experimental tests of this theory. We sampled the effect of long-term exclusion of large vertebrate herbivores on the prevalence of infection by a group of aphid-vectored viruses that infect grasses (barley and cereal yellow dwarf viruses) in an oak savannah in central California. We found that pathogen prevalence was approximately 4-fold higher in the presence of consumers than in areas where they were excluded. Vertebrate consumers did not directly alter infection rates by this aphid vectored pathogen group, but rather increased infection risk by increasing the relative abundance of highly-competent hosts in the grassland community. This large-scale experiment, measuring changes in host abundance and infection risk in response to altered consumption rates, confirms theoretical predictions that consumers can indirectly increase infection risk by altering the composition of whole communities. Most importantly, these results demonstrate that, even in complex natural communities, alterations to food web composition such as consumer invasion or extinction can lead to significant impacts that cascade throughout entire communities, including changes in infection risk. PMID- 19126682 TI - Evolution of F-box genes in plants: different modes of sequence divergence and their relationships with functional diversification. AB - F-box proteins are substrate-recognition components of the Skp1-Rbx1-Cul1-F-box protein (SCF) ubiquitin ligases. In plants, F-box genes form one of the largest multigene superfamilies and control many important biological functions. However, it is unclear how and why plants have acquired a large number of F-box genes. Here we identified 692, 337, and 779 F-box genes in Arabidopsis, poplar and rice, respectively, and studied their phylogenetic relationships and evolutionary patterns. We found that the plant F-box superfamily can be divided into 42 families, each of which has a distinct domain organization. We also estimated the number of ancestral genes for each family and identified highly conservative versus divergent families. In conservative families, there has been little or no change in the number of genes since the divergence between eudicots and monocots approximately 145 million years ago. In divergent families, however, the numbers have increased dramatically during the same period. In two cases, the numbers of genes in extant species are >100 times greater than that in the most recent common ancestor (MRCA) of the three species. Proteins encoded by highly conservative genes always have the same domain organization, suggesting that they interact with the same or similar substrates. In contrast, proteins of rapidly duplicating genes sometimes have quite different domain structures, mainly caused by unusually frequent shifts of exon-intron boundaries and/or frameshift mutations. Our results indicate that different F-box families, or different clusters of the same family, have experienced dramatically different modes of sequence divergence, apparently having resulted in adaptive changes in function. PMID- 19126683 TI - Cancer cell angiogenic capability is regulated by 3D culture and integrin engagement. AB - Three-dimensional culture alters cancer cell signaling; however, the underlying mechanisms and importance of these changes on tumor vascularization remain unclear. A hydrogel system was used to examine the role of the transition from 2D to 3D culture, with and without integrin engagement, on cancer cell angiogenic capability. Three-dimensional culture recreated tumor microenvironmental cues and led to enhanced interleukin 8 (IL-8) secretion that depended on integrin engagement with adhesion peptides coupled to the polymer. In contrast, vascular endothelial growth factor (VEGF) secretion was unaffected by 3D culture with or without substrate adhesion. IL-8 diffused greater distances and was present in higher concentrations in the systemic circulation, relative to VEGF. Implantation of a polymeric IL-8 delivery system into GFP bone marrow-transplanted mice revealed that localized IL-8 up-regulation was critical to both the local and systemic control of tumor vascularization in vivo. In summary, 3D integrin engagement within tumor microenvironments regulates cancer cell angiogenic signaling, and controlled local and systemic blockade of both IL-8 and VEGF signaling may improve antiangiogenic therapies. PMID- 19126684 TI - Glucocorticoid receptor interaction with TrkB promotes BDNF-triggered PLC-gamma signaling for glutamate release via a glutamate transporter. AB - An increase in glucocorticoid levels and down-regulation of BDNF (brain-derived neurotrophic factor) are supposed to be involved in the pathophysiology of depressive disorders. However, possible crosstalk between glucocorticoid- and BDNF-mediated neuronal functions in the CNS has not been elucidated. Here, we examined whether chronic glucocorticoid exposure influences BDNF-triggered intracellular signaling for glutamate release via a glutamate transporter. We found that chronic exposure to dexamethasone (DEX, a synthetic glucocorticoid) suppressed BDNF-induced glutamate release via weakening the activation of the PLC gamma (phospholipase C-gamma)/Ca(2+) system in cultured cortical neurons. We demonstrated that the GR (glucocorticoid receptor) interacts with receptor tyrosine kinase for BDNF (TrkB). Following DEX treatment, TrkB-GR interaction was reduced due to the decline in GR expression. Corticosterone, a natural glucocorticoid, also reduced TrkB-GR interaction, BDNF-stimulated PLC-gamma, and BDNF-triggered glutamate release. Interestingly, BDNF-dependent binding of PLC gamma to TrkB was diminished by DEX. SiRNA transfection to induce a decrease in endogenous GR mimicked the inhibitory action of DEX. Conversely, DEX-inhibited BDNF-activated PLC-gamma signaling for glutamate release was recovered by GR overexpression. We propose that TrkB-GR interaction plays a critical role in the BDNF-stimulated PLC-gamma pathway, which is required for glutamate release, and the decrease in TrkB-GR interaction caused by chronic exposure to glucocorticoids results in the suppression of BDNF-mediated neurotransmitter release via a glutamate transporter. PMID- 19126685 TI - Regulatory effects of ferritin on angiogenesis. AB - Angiogenesis, the synthesis of new blood vessels from preexisting vessels, plays a critical role in normal wound healing and tumor growth. HKa (cleaved high molecular weight kininogen) is an endogenous inhibitor of angiogenesis formed by the cleavage of kininogen on endothelial cells. Ferritin is a protein principally known for its central role in iron storage. Here, we demonstrate that ferritin binds to HKa with high affinity (K(d) 13 nM). Further, ferritin antagonizes the antiangiogenic effects of HKa, enhancing the migration, assembly, and survival of HKa-treated endothelial cells. Effects of ferritin were independent of its iron content. Peptide mapping revealed that ferritin binds to a 22-aa subdomain of HKa that is critical to its antiangiogenic activity. In vivo, ferritin opposed HKa's antiangiogenic effects in a human prostate cancer xenograft, restoring tumor dependent vessel growth. Ferritin-mediated regulation of angiogenesis represents a new angiogenic regulatory pathway, and identifies a new role for ferritin in cell biology. PMID- 19126686 TI - Soluble epoxide hydrolase plays an essential role in angiotensin II-induced cardiac hypertrophy. AB - Pathophysiological cardiac hypertrophy is one of the most common causes of heart failure. Epoxyeicosatrienoic acids, hydrolyzed and degraded by soluble epoxide hydrolase (sEH), can function as endothelium-derived hyperpolarizing factors to induce dilation of coronary arteries and thus are cardioprotective. In this study, we investigated the role of sEH in two rodent models of angiotensin II (Ang II)-induced cardiac hypertrophy. The protein level of sEH was elevated in the heart of both spontaneously hypertensive rats and Ang II-infused Wistar rats. Blocking the Ang II type 1 receptor with losartan could abolish this induction. Administration of a potent sEH inhibitor (sEHI) prevented the pathogenesis of the Ang II-induced hypertrophy, as demonstrated by decreased left-ventricular hypertrophy assessed by echocardiography, reduced cardiomyocyte size, and attenuated expression of hypertrophy markers, including atrial natriuretic factor and beta-myosin heavy chain. Because sEH elevation was not observed in exercise- or norepinephrine-induced hypertrophy, the sEH induction was closely associated with Ang II-induced hypertrophy. In vitro, Ang II upregulated sEH and hypertrophy markers in neonatal cardiomyocytes isolated from rat and mouse. Expression of these marker genes was elevated with adenovirus-mediated sEH overexpression but decreased with sEHI treatment. These results were supported by studies in neonatal cardiomyocytes from sEH(-/-) mice. Our results suggest that sEH is specifically upregulated by Ang II, which directly mediates Ang II-induced cardiac hypertrophy. Thus, pharmacological inhibition of sEH would be a useful approach to prevent and treat Ang II-induced cardiac hypertrophy. PMID- 19126688 TI - Splendor in the grasses. PMID- 19126689 TI - A recommendation for naming transcription factor proteins in the grasses. PMID- 19126690 TI - Increasing crop productivity to meet global needs for feed, food, and fuel. PMID- 19126692 TI - Revolutionary times in our understanding of cell wall biosynthesis and remodeling in the grasses. PMID- 19126691 TI - The development of endosperm in grasses. PMID- 19126693 TI - Translational biology: from Arabidopsis flowers to grass inflorescence architecture. PMID- 19126694 TI - Hormonal regulation of branching in grasses. PMID- 19126695 TI - Mechanisms of floral induction in grasses: something borrowed, something new. PMID- 19126696 TI - Genes and mutations underlying domestication transitions in grasses. PMID- 19126697 TI - Genetic control of carbon partitioning in grasses: roles of sucrose transporters and tie-dyed loci in phloem loading. PMID- 19126698 TI - Integrating phylogeny into studies of C4 variation in the grasses. PMID- 19126699 TI - Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses. PMID- 19126700 TI - Indirect defense responses to herbivory in grasses. PMID- 19126701 TI - For blighted waves of grain: Fusarium graminearum in the postgenomics era. PMID- 19126702 TI - Synergy of two reference genomes for the grass family. PMID- 19126704 TI - Genomic and genetic database resources for the grasses. PMID- 19126703 TI - Comparative genomics of grasses promises a bountiful harvest. PMID- 19126705 TI - Foxtail millet: a sequence-driven grass model system. PMID- 19126706 TI - The international barley sequencing consortium--at the threshold of efficient access to the barley genome. PMID- 19126707 TI - Cereal germplasm resources. PMID- 19126708 TI - Resources for virus-induced gene silencing in the grasses. PMID- 19126709 TI - TILLING in grass species. PMID- 19126710 TI - Mutant resources in rice for functional genomics of the grasses. PMID- 19126714 TI - Lactobacillus fabifermentans sp. nov. and Lactobacillus cacaonum sp. nov., isolated from Ghanaian cocoa fermentations. AB - Two Gram-positive bacterial strains, LMG 24284T and LMG 24285T, were isolated from different spontaneous cocoa bean heap fermentations in Ghana. Analysis of their 16S rRNA gene sequences indicated that they were members of the Lactobacillus plantarum and Lactobacillus salivarius species groups, respectively. DNA-DNA hybridization experiments with their nearest phylogenetic neighbours demonstrated that both strains represented novel species that could be differentiated from their nearest neighbours by pheS sequence analysis, whole cell protein electrophoresis, fluorescent amplified fragment length polymorphism analysis and biochemical characterization. Therefore, two novel Lactobacillus species are proposed, Lactobacillus fabifermentans sp. nov. (type strain LMG 24284T =DSM 21115T) and Lactobacillus cacaonum sp. nov. (type strain LMG 24285T =DSM 21116T). PMID- 19126715 TI - Paenibacillus harenae sp. nov., isolated from desert sand in China. AB - A Gram-positive, endospore-forming, rod-shaped bacterium, designated strain B519T, was isolated from a desert sand sample of Gansu Province, China. Strain B519T was strictly aerobic and cells were motile by means of peritrichous flagella. The strain grew optimally at 32-35 degrees C and pH 6.5-7.0. Chemotaxonomic data supported the affiliation of the new isolate to the genus Paenibacillus, including menaquinone-7 (MK-7) as the major isoprenoid quinone, DNA G+C content of 49.9 mol%, cell-wall type A1gamma (meso-diaminopimelic acid as the diagnostic diamino acid) and anteiso-C(15 : 0), iso-C(15 : 0), C(16 : 0) and iso-C(16 : 0) as the major fatty acids. Comparative 16S rRNA gene sequence analysis showed that strain B519T was most closely related to Paenibacillus alkaliterrae KSL-134T (98.0 % similarity). DNA-DNA relatedness between strain B519T and P. alkaliterrae KSL-134T was about 12.3 %. On the basis of phenotypic characteristics and molecular properties, strain B519T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus harenae sp. nov. is proposed. The type strain is B519T (=KCTC 3951T =DSM 16969T). PMID- 19126716 TI - Leifsonia kribbensis sp. nov., isolated from soil. AB - A yellow-pigmented actinobacterium, designated strain MSL-13T, was isolated from a soil sample collected from Bigeum Island, Republic of Korea, and its taxonomic position was determined by using a polyphasic approach. Strain MSL-13T showed phenotypic and chemotaxonomic properties consistent with its classification in the genus Leifsonia. 16S rRNA gene sequence analysis of strain MSL-13T with sequences from Leifsonia naganoensis DB103T, Leifsonia aquatica DSM 20146T, Leifsonia xyli subsp. cynodontis JCM 9733T, Leifsonia poae VKM Ac-1401T and Leifsonia shinshuensis DB102T revealed similarities of 96.22, 96.19, 95.77, 95.44 and 95.37 %, respectively, with differences of 39-65 nt among 1483 total nucleotides aligned. Based on differences in phenotypic and genotypic characteristics, strain MSL-13T (=KCTC 19267T =DSM 19272T) is designated as the type strain of a novel species of the genus Leifsonia, for which the name Leifsonia kribbensis sp. nov. is proposed. PMID- 19126717 TI - Methylobacterium phyllosphaerae sp. nov., a pink-pigmented, facultative methylotroph from the phyllosphere of rice. AB - A pink-pigmented, aerobic, facultatively methylotrophic bacterial strain, CBMB27T, isolated from leaf tissues of rice (Oryza sativa L. 'Dong-Jin'), was analysed using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence-based phylogenetic analysis placed the strain in a clade with the species Methylobacterium oryzae, Methylobacterium fujisawaense and Methylobacterium mesophilicum; strain CBMB27T showed sequence similarities of 98.3, 98.5 and 97.3 %, respectively, to the type strains of these three species. DNA-DNA hybridization experiments revealed low levels (<38 %) of DNA-DNA relatedness between strain CBMB27T and its closest relatives. The sequence of the 1-aminocyclopropane-1-carboxylate deaminase gene (acdS) in strain CBMB27T differed from those of close relatives. The major fatty acid of the isolate was C(18 : 1)omega7c and the G+C content of the genomic DNA was 66.8 mol%. Based on the results of 16S rRNA gene sequence analysis, DNA-DNA hybridization, and physiological and biochemical characterization, which enabled the isolate to be differentiated from all recognized species of the genus Methylobacterium, it was concluded that strain CBMB27T represents a novel species in the genus Methylobacterium for which the name Methylobacterium phyllosphaerae sp. nov. is proposed (type strain CBMB27T =LMG 24361T =KACC 11716T =DSM 19779T). PMID- 19126719 TI - Corynebacterium ulceribovis sp. nov., isolated from the skin of the udder of a cow with a profound ulceration. AB - A coryneform bacterium isolated from a cow with a profound ulceration was characterized by using phenotypic and molecular taxonomic methods. Chemotaxonomic investigations revealed the presence of cell-wall chemotype IV and short-chain mycolic acids consistent with the genus Corynebacterium. Comparative 16S rRNA gene sequencing showed that the organism formed a hitherto unknown subline within the genus Corynebacterium. Sequence divergence values of greater than 3.6 % from recognized Corynebacterium species, together with phenotypic differences, showed that the unidentified bacterium represents a new member of this genus. On the basis of phenotypic and phylogenetic considerations, it is proposed that the novel bacterium be classified as representing a novel species of the genus Corynebacterium, with the name Corynebacterium ulceribovis sp. nov. The type strain is IMMIB L-1395T (=DSM 45146T=CCUG 55727T). PMID- 19126718 TI - Paraliobacillus quinghaiensis sp. nov., isolated from salt-lake sediment in China. AB - A Gram-positive, moderately halophilic, endospore-forming, catalase- and oxidase positive, obligately aerobic bacterium, designated strain YIM-C158T, was isolated from sediment of a salt lake in the Qaidam Basin, north-west China. Cells were motile with peritrichous flagella and rod-shaped, with meso-diaminopimelic acid in the cell-wall peptidoglycan. Strain YIM-C158T grew in the presence of 1-20 % (w/v) NaCl and pH 6.0-10.0, with optimum growth at 5 % (w/v) NaCl and pH 8.0. The strain grew at 4-50 degrees C, with optimum growth at 37 degrees C. The major cellular fatty acids were anteiso-C(15 : 0), iso-C(14 : 0), C(16 : 0), anteiso C(17 : 0), iso-C(16 : 0), C(16 : 1)omega7c alcohol and C(16 : 1)omega11c. Strain YIM-C158T contained menaquinone MK-7 as the sole respiratory quinone and phosphatidylmethylethanolamine and phosphatidylcholine as the polar lipids. The genomic DNA G+C content was 39.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM-C158T was most closely related to Paraliobacillus ryukyuensis DSM 15140T (96.8 % similarity), and the two strains formed a distinct branch in the phylogenetic tree. The level of DNA-DNA relatedness between the two strains was 15.6 %. The combination of phylogenetic analysis, phenotypic characteristics, chemotaxonomic differences and DNA-DNA hybridization data supported the view that strain YIM-C158T represents a novel species of the genus Paraliobacillus, for which the name Paraliobacillus quinghaiensis sp. nov. is proposed. The type strain is YIM-C158T (=DSM 17857T=CGMCC 1.6333T). PMID- 19126720 TI - Pseudomonas sabulinigri sp. nov., isolated from black beach sand. AB - A novel Gram-negative, aerobic, motile, short rod-shaped bacterium, designated J64T, was isolated from black sand collected from Soesoggak, Jeju Island, Korea. Cells grew at 4-37 degrees C, at pH 5.5-10.0 and with 0-10 % NaCl. The strain was found to be oxidase- and catalase-positive. Phylogenetic analyses showed that strain J64T belongs to the genus Pseudomonas, forming a monophyletic group with Pseudomonas pachastrellae, Pseudomonas pertucinogena and 'Pseudomonas denitrificans'. The 16S rRNA gene sequence similarity between strain J64T and type strains of all Pseudomonas species with validly published names was below 96.6 %. Low levels of DNA-DNA relatedness were found with respect to type strains of P. pachastrellae and P. pertucinogena, supporting the classification of strain J64T within a novel species of the genus Pseudomonas. Strain J64T contained C(18 : 1)omega7c (37.2 %), C(16 : 0) (20.4 %), summed feature 3 (17.4 %; comprising iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) and C(12 : 0) (7.6 %) as major cellular fatty acids. On the basis of the phenotypic and phylogenetic data, strain J64T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas sabulinigri sp. nov. is proposed. The type strain is J64T (=KCTC 22137T =JCM 14963T). PMID- 19126721 TI - Nocardioides basaltis sp. nov., isolated from black beach sand. AB - A novel Gram-positive, aerobic, short-rod-shaped bacterium, designated strain J112T, was isolated from black sand collected from Soesoggak, Jeju Island, Korea. The strain was found to be oxidase-negative and catalase-positive. Cells grew at 10-37 degrees C, at pH 5.5-8.0 and with 1-10 % NaCl. Growth occurred on marine agar but not on R2A or trypticase soy agar. A phylogenetic analysis based on 16S rRNA gene sequences showed that the strain belongs to the radiation of the genus Nocardioides. Strain J112T shared the highest 16S rRNA gene sequence similarities with Nocardioides marinisabuli SBS-12T (99.2 %), Nocardioides terrigena DS-17T (97.3 %), Nocardioides kribbensis KCTC 19038T (97.1 %) and type strains of other Nocardioides species with validly published names (<97 %). The DNA-DNA hybridization values between strain J112T and the three most closely related strains were low enough to justify the assignment of this strain to a novel species. On the basis of these phenotypic, phylogenetic and chemotaxonomic data, strain J112T represents a novel species of the genus Nocardioides, for which the name Nocardioides basaltis sp. nov. is proposed. The type strain is J112T (=KCTC 19365T=JCM 14945T). PMID- 19126722 TI - Lutimaribacter saemankumensis gen. nov., sp. nov., isolated from a tidal flat of the Yellow Sea. AB - A Gram-negative, non-motile, rod-shaped bacterial strain, designated SMK-117T, belonging to the Alphaproteobacteria, was isolated from a tidal flat of the Yellow Sea, Korea, and was subjected to a polyphasic taxonomic study. Strain SMK 117T grew optimally at pH 7.0-8.0 and 30 degrees C in the presence of 2 % (w/v) NaCl. Neighbour-joining and maximum-likelihood phylogenetic trees based on 16S rRNA gene sequences showed that strain SMK-117T clustered with Maritimibacter alkaliphilus HTCC2654T, with which it exhibited a sequence similarity of 95.3 %. Strain SMK-117T contained Q-10 as the predominant ubiquinone and C(18 : 1)omega7c and 11-methyl C(18 : 1)omega7c as the major fatty acids. The major polar lipids were phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminolipid and two unidentified phospholipids. The DNA G+C content was 63.5 mol%. Strain SMK-117T was differentiated from members of the genera Maritimibacter and Oceanicola on the basis of differences in the fatty acid and polar lipid profiles. The phenotypic, chemotaxonomic and phylogenetic data indicated that strain SMK-117T represents a novel genus and species, for which the name Lutimaribacter saemankumensis gen. nov., sp. nov. is proposed. The type strain of Lutimaribacter saemankumensis is SMK-117T (=KCTC 22244T =CCUG 55760T). PMID- 19126723 TI - Pseudidiomarina marina sp. nov. and Pseudidiomarina tainanensis sp. nov. and reclassification of Idiomarina homiensis and Idiomarina salinarum as Pseudidiomarina homiensis comb. nov. and Pseudidiomarina salinarum comb. nov., respectively. AB - Two Gram-negative strains of heterotrophic, aerobic, marine bacteria, designated PIM1T and PIN1T, were isolated from seawater samples collected from the shallow coastal region of An-Ping Harbour, Tainan, Taiwan. Cells grown in broth cultures were straight rods and non-motile. The two isolates required NaCl for growth and grew optimally at 30-35 degrees C and 2-5 % NaCl. They grew aerobically and were not capable of anaerobic growth by fermentation of glucose or other carbohydrates. The cellular fatty acids were predominantly iso-branched, with iso C(15 : 0) (17.0-21.4 %), iso-C(17 : 0) (18.2-21.0 %) and iso-C(17 : 1)omega9c (15.7-16.6 %) as the most abundant components. The predominant isoprenoid quinone was Q-8 (95.2-97.1 %). Strains PIM1T and PIN1T had DNA G+C contents of 46.6 and 46.9 mol%, respectively. Phylogeny based on 16S rRNA gene sequences and DNA-DNA hybridization, together with data from physiological, morphological and chemotaxonomic characterizations, indicated that the two isolates should be classified as representatives of two novel species of the genus Pseudidiomarina of the family Idiomarinaceae, for which the names Pseudidiomarina marina sp. nov. (type strain PIM1T=BCRC 17749T=JCM 15083T) and Pseudidiomarina tainanensis sp. nov. (type strain PIN1T=BCRC 17750T=JCM 15084T) are proposed. In addition, based on the characterization data obtained in this study, it is proposed that Idiomarina homiensis and Idiomarina salinarum should be reclassified as Pseudidiomarina homiensis comb. nov. and Pseudidiomarina salinarum comb. nov., respectively. PMID- 19126724 TI - Dyadobacter alkalitolerans sp. nov., isolated from desert sand. AB - A polyphasic taxonomic approach was used to study strain 12116T, which was isolated from desert sand in Xinjiang, China. Cells of the isolate were Gram negative, non-motile, rod-shaped and aerobic. MK-7 was the predominant respiratory menaquinone. The DNA G+C content was 46.3 mol%. A phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate was most closely related to the members of the genus Dyadobacter, with similarities ranging from 94.7 to 96.6 %. Therefore, on the basis of genotypic and phenotypic data, strain 12116T represents a novel species of the genus Dyadobacter, for which the name Dyadobacter alkalitolerans sp. nov. is proposed. The type strain is 12116T (=CCTCC AB 207176T =NRRL B-51268T). PMID- 19126725 TI - Lewinella antarctica sp. nov., a marine bacterium isolated from Antarctic seawater. AB - A marine bacterium, designated IMCC3223T, was isolated from Antarctic coastal seawater and subjected to a polyphasic taxonomic investigation. Cells of the strain were Gram-negative, short to elongated rods that were orange-coloured but negative for flexirubin-type pigments, obligately aerobic, chemoheterotrophic and devoid of gliding or flagellar motility. On the basis of 16S rRNA gene sequence comparisons, the strain was closely related to members of the genus Lewinella (86.3-93.1 %) of the family 'Saprospiraceae' in the phylum Bacteroidetes; the most closely related species was Lewinella lutea (93.1 % similarity to the type strain). The DNA G+C content of the strain was 50.3 mol% and the major respiratory quinone was MK-7. The predominant cellular fatty acids were C(16 : 1)omega7c/iso-C(15 : 0) 2-OH (39.4 %), iso-C(15 : 0) (15.9 %), C(17 : 1)omega7c (10.5 %) and C(15 : 1)omega6c (8.9 %). 16S rRNA gene sequence analyses and the phenotypic and chemotaxonomic tests allowed the differentiation of strain IMCC3223T from all recognized species of the genus Lewinella. Therefore, strain IMCC3223T represents a novel species of the genus Lewinella, for which the name Lewinella antarctica sp. nov. is proposed. The type strain is IMCC3223T (=KCCM 42688T =NBRC 103142T). PMID- 19126726 TI - Pedobacter daechungensis sp. nov., from freshwater lake sediment in South Korea. AB - A novel bacterial strain, designated Dae 13T, was isolated from sediment from a freshwater lake in Daejeon, South Korea, and was characterized taxonomically by using a polyphasic approach. The isolate was Gram-negative, aerobic, non-motile, non-spore-forming and rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belonged to the genus Pedobacter in the family Sphingobacteriaceae but was clearly separate from established species of this genus. The 16S rRNA gene sequence similarities between strain Dae 13(T) and type strains of Pedobacter species with validly published names ranged from 91.6 to 97.5 %. The G+C content of the genomic DNA was 33.8 mol%. Chemotaxonomic data, i.e. the presence of MK-7 as the major menaquinone and iso-C(15 : 0), C(16 : 0) and summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) as the major fatty acids, supported the affiliation of strain Dae 13T to the genus Pedobacter. However, the results of physiological and biochemical tests allowed phenotypic differentiation of the isolate with respect to Pedobacter species with validly published names. Therefore, strain Dae 13T represents a novel species within the genus Pedobacter, for which the name Pedobacter daechungensis sp. nov. is proposed. The type strain is Dae 13T (=KCTC 12637T=LMG 23489T). PMID- 19126727 TI - Glaciecola lipolytica sp. nov., isolated from seawater near Tianjin city, China. AB - A Gram-negative, non-spore-forming, rod-shaped bacterium, designated strain E3T, which was motile with a polar flagellum, was isolated from coastal surface seawater near Tianjin city, China. The taxonomy of strain E3T was studied by phenotypic, chemotaxonomic and phylogenetic methods. Strain E3T forms non pigmented colonies with a butyrous-mucoid consistency on marine 2216 agar. It grows optimally at 30 degrees C, in the presence of 2.0 % (w/v) NaCl and at pH 7.6. Strain E3T is positive for catalase and oxidase and hydrolysis of lipid and reduces nitrate to N2 gas. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain E3T belongs to the Gammaproteobacteria and is grouped in the genus Glaciecola, with highest sequence similarity to the type strains of Glaciecola chathamensis (94.6 %), G. agarilytica (94.4 %) and G. mesophila (94.4 %). Strain E3T contained summed feature 3 (C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, 34.5 %), C(18 : 1)omega7c (17.3 %) and C(16 : 0) (17 %) as the major fatty acids and phosphatidylethanolamine and phosphatidylglycerol as the main phospholipids. The DNA G+C content was 40.8 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain E3T is considered to represent a novel species of the genus Glaciecola, for which the name Glaciecola lipolytica sp. nov. is proposed. The type strain is E3T (=CGMCC 1.7001T =JCM 15139T). PMID- 19126728 TI - Hymenobacter deserti sp. nov., isolated from the desert of Xinjiang, China. AB - A Gram-negative, rod-shaped, non-motile and pink-pigmented bacterium, designated strain ZLB-3T, was isolated from a desert soil sample collected from Xinjiang Province, China, and characterized by using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belongs to the phylum Bacteroidetes and is related to the genus Hymenobacter. 16S rRNA gene sequence similarities between strain ZLB-3T and the type strains of Hymenobacter species with validly published names ranged from 90.0 to 94.6 %. No other taxa in the phylum Bacteroidetes showed more than 90 % sequence similarity to the isolate. The strain contained MK-7 as the predominant menaquinone. The major fatty acids were iso-C(15 : 0) (27.8 %), anteiso-C(17 : 1) B/iso-C(17 : 1) I (25.2 %), iso-C(17 : 0) 3-OH (9.6 %) and C(16 : 1)omega7c/iso-C(15 : 0) 2-OH (8.5 %). Phosphatidylethanolamine, two unknown aminophospholipids, an unknown aminolipid, an unknown glycolipid and six unknown polar lipids were detected in the polar lipid profile. The DNA G+C content was 58.6 mol%. These chemotaxonomic data supported the affiliation of strain ZLB-3T to the genus Hymenobacter. However, the results of physiological and biochemical tests allowed phenotypic differentiation of the isolate from recognized Hymenobacter species. On the basis of the evidence presented, it is proposed that strain ZLB-3T represents a novel species, Hymenobacter deserti sp. nov. The type strain is ZLB-3T (=CCTCC AB 207171T =NRRL B-51267T). PMID- 19126729 TI - Lactobacillus nodensis sp. nov., isolated from rice bran. AB - Gram-positive, rod-shaped, non-motile lactic acid bacteria, strains iz4b-1T, iz4b 2 and iz4c-1, were isolated in an attempt to study the composition of the Lactobacillus flora of Japanese pickles. Analysis of their 16S rRNA gene sequences revealed that the strains clustered in the Lactobacillus alimentarius group, and comparatively high similarities were shown to 'Lactobacillus tucceti' CECT 5920 (97.0 %), Lactobacillus versmoldensis KU-3T (96.4 %) and Lactobacillus nantensis LP33T (94.4 %). DNA-DNA hybridization assays clearly revealed that the isolates represented a novel taxon. The DNA G+C content was 40.6 mol% and the peptidoglycan type was L-Lys-D-Asp. Thus, these isolates represent a novel Lactobacillus species, for which the name Lactobacillus nodensis sp. nov. is proposed. The type strain is iz4b-1T (=DSM 19682T =JCM 14932T). PMID- 19126730 TI - Description of Patulibacter americanus sp. nov., isolated from biological soil crusts, emended description of the genus Patulibacter Takahashi et al. 2006 and proposal of Solirubrobacterales ord. nov. and Thermoleophilales ord. nov. AB - Two bacterial strains, CP153-3 and CP177-2T, were isolated from biological soil crusts on the Colorado Plateau, USA, and assigned to the genus Patulibacter, according to 16S rRNA gene sequence analysis and phenotypic characteristics. The strains were Gram-positive, aerobic and psychrotolerant and showed positive catalase and negative oxidase reactions. Cells were motile in young cultures, motility consisting of subtle jerking motions, but no flagella could be detected. The strains shared high 16S rRNA gene sequence similarity (99.7 %) and DNA-DNA relatedness (99 %). They contained meso-diaminopimelic acid in the cell wall, the major menaquinone was MK-7(H2) and the predominant fatty acids were C(18 : 1)omega9c and iso-C(15 : 0). A low DNA-DNA hybridization value of 20 % with the type strain of the type species of the genus Patulibacter, Patulibacter minatonensis, as well as phenotypic differences, indicated that the isolates belong to a novel species, for which the name Patulibacter americanus sp. nov. is proposed. Strain CP177-2T (=ATCC BAA-1038(T) =DSM 16676T) was chosen as the type strain. An emended description of the genus Patulibacter is also presented. Further, two new orders are created to accommodate several families previously classified in the order Rubrobacterales Rainey et al. 1997. The first, Solirubrobacterales ord. nov., encompasses the families Solirubrobacteraceae, Conexibacteraceae and Patulibacteraceae, and the second, Thermoleophilales ord. nov., includes the family Thermoleophilaceae. PMID- 19126731 TI - Caloramator australicus sp. nov., a thermophilic, anaerobic bacterium from the Great Artesian Basin of Australia. AB - A strictly anaerobic, thermophilic bacterium, designated strain RC3T, was isolated from microbial mats colonizing thermal waters of a run-off channel formed by free-flowing waters from a bore well (registered no. 17263) of the Great Artesian Basin, Australia. The slightly curved rods (2.5-4.2x0.8-1.0 microm) of strain RC3T stained Gram-positive and grew optimally in tryptone-yeast extract-glucose medium at 60 degrees C (range 45-70 degrees C) and pH 7 (range pH 5-9). Strain RC3T grew poorly on yeast extract (0.2 %) but did not grow on tryptone (0.2 %) as a sole carbon source; yeast extract was required for growth on other energy sources, which included glucose, fructose, galactose, xylose, maltose, sucrose, raffinose, mannose, cellobiose, cellulose, starch, amylopectin, xylan, peptone, amyl media (Research Achievement), threonine and pyruvate but did not include arabinose, ribose, lactose, CM-cellulose, myo-inositol, mannitol, chitin, casein, formate, acetate, succinate, propionate, lactate, benzoate, glycerol, ethanol, Casamino acids, arginine, alanine, serine, glycine, glutamine, leucine, isoleucine, methionine or aspartate. The end products of glucose fermentation were ethanol and acetate. In the presence of 0.2 % yeast extract, iron(III), manganese(IV) and elemental sulfur were reduced but not sulfate, sulfite, thiosulfate, nitrate or nitrite. Iron(III) was also reduced in the presence of peptone, tryptone, amyl media, threonine and glycerol but not chitin, xylan, pectin, starch, pyruvate, acetate, benzoate, lactate, propionate, succinate, inositol, ethanol, mannitol, arginine, glutamine or serine. Strain RC3T was not able to utilize molecular hydrogen and/or carbon dioxide in the presence or absence of iron(III). In the presence of iron(III) and glycerol, increased concentrations of Fe(II) corresponded to increased cell numbers, demonstrating that strain RC3(T) was able to conserve energy to support growth from the reduction of Fe(III) to Fe(II). Chloramphenicol, streptomycin, tetracycline, penicillin and ampicillin and NaCl concentrations greater than 2 % inhibited growth. The G+C content of the DNA was 34+/-1 mol% as determined by the thermal denaturation (Tm) method. 16S rRNA gene sequence analysis indicated that strain RC3T was affiliated to Caloramator fervidus (95.8 % similarity to the type strain) and to other Caloramator species (average similarity of 91.6 %) within the phylum Firmicutes. On the basis of phylogenetic and phenotypic characteristics, it is proposed that strain RC3T should be classified in the genus Caloramator as a representative of a novel species, Caloramator australicus sp. nov. The type strain is RC3T (=JCM 1508T =KCTC 5601T). PMID- 19126732 TI - Taxon K, a complex within the Burkholderia cepacia complex, comprises at least two novel species, Burkholderia contaminans sp. nov. and Burkholderia lata sp. nov. AB - The aim of the present study was to re-examine the taxonomic position and structure of taxon K (also known as group K) within the Burkholderia cepacia complex (Bcc). For this purpose, a representative set of strains was examined by a traditional polyphasic taxonomic approach, by multilocus sequence typing (MLST) analysis and by analysis of available whole-genome sequences. Analysis of the recA gene sequence revealed three different lineages, designated recA-I, recA-II and recA-III. DNA-DNA hybridization experiments demonstrated that recA-I and recA II isolates each represented a single novel species. However, DNA-DNA hybridization values of recA-II strains towards recA-III strains and among recA III strains were at the threshold level for species delineation. By MLST, recA-I isolates were clearly distinguished from the others and represented a distinct lineage referred to as MLST-I, whereas recA-II and recA-III isolates formed a second MLST lineage referred to as MLST-II. A divergence value of 3.5 % was obtained when MLST-I was compared with MLST-II. The internal level of concatenated sequence divergence within MLST-I and MLST-II was 1.4 and 2.7 %, respectively; by comparison with the level of concatenated sequence divergence in established Bcc species, these data demonstrate that the MLST-I and MLST-II lineages represent two distinct species within the Bcc. The latter conclusion was supported by comparison of the whole-genome average nucleotide identity (ANI) level of MLST-I and MLST-II strains with strains of established Bcc species and by a whole-genome-based phylogenetic analysis. We formally propose to classify taxon K bacteria from the MLST-I and MLST-II lineages as Burkholderia contaminans sp. nov. (with strain J2956T =LMG 23361T =CCUG 55526T as the type strain) and Burkholderia lata sp. nov. (with strain 383T =ATCC 17760T =LMG 22485T =CCUG 55525T as the type strain), respectively. The MLST approach was confirmed as a valuable instrument in polyphasic taxonomic studies; more importantly, the cumulative data for about 1000 Bcc isolates analysed demonstrate that the 3 % concatenated sequence divergence level correlates with the 70 % DNA-DNA hybridization or 95 % whole-genome ANI threshold levels for species delineation. PMID- 19126733 TI - Description of seven candidate species affiliated with the phylum Actinobacteria, representing planktonic freshwater bacteria. AB - Actinobacteria comprise a substantial fraction of the bacterioplankton in freshwater lakes and streams. Numerous cultivation-independent investigations have retrieved actinobacterial 16S rRNA gene sequences from such habitats. The taxa detected in freshwater habitats are usually absent from terrestrial and marine systems. So far, none of the indigenous freshwater lineages is represented by a taxon with a validly published name. The seven organisms for which Candidatus status is described here were isolated from freshwater lakes and ponds located in tropical, subtropical and temperate climatic zones. Phylogenetic analyses demonstrated that they are affiliated with one of the actinobacterial lineages indigenous to freshwater bacterioplankton. The seven novel taxa could only be cultivated to date as mixed cultures that also contain non actinobacterial strains. Due to the lack of pure cultures, I propose to establish the candidate species 'Candidatus Planktoluna difficilis', 'Candidatus Aquiluna rubra', 'Candidatus Flaviluna lacus', 'Candidatus Rhodoluna limnophila', 'Candidatus Rhodoluna planktonica', 'Candidatus Rhodoluna lacicola' and 'Candidatus Limnoluna rubra' for these taxa. PMID- 19126734 TI - Acinetobacter beijerinckii sp. nov. and Acinetobacter gyllenbergii sp. nov., haemolytic organisms isolated from humans. AB - The taxonomic status of 24 haemolytic, non-glucose acidifying Acinetobacter strains that did not belong to any previously described species was investigated by means of a polyphasic approach. Using AFLP fingerprinting, amplified rDNA restriction analysis and phenotypic characterization, the strains were classified into two phenetically coherent groups (comprising 15 and 9 strains) that were distinct from each other and from all known Acinetobacter species. Confirmation that these groups formed two separate lineages within the genus Acinetobacter was obtained from comparative analysis of partial sequences of the gene encoding the beta-subunit of RNA polymerase in all strains and also from 16S rRNA gene sequence analysis of representative strains. Previously published DNA-DNA reassociation data for some of the strains used also supported the species rank for both groups, for which the names Acinetobacter beijerinckii sp. nov. and Acinetobacter gyllenbergii sp. nov. are proposed. The strains of A. beijerinckii sp. nov. originated from human and animal specimens and from various environmental sources, whereas those of A. gyllenbergii sp. nov. were isolated exclusively from human clinical specimens. The phenotypic characteristics most useful for the differentiation of these species from other Acinetobacter species that comprise haemolytic strains were the inability of A. beijerinckii sp. nov. to grow on l-arginine and the ability of A. gyllenbergii sp. nov. to grow on azelate. The type strain of A. beijerinckii sp. nov. is NIPH 838T (=LUH 4759T=CCUG 51249T=CCM 7266T=58aT) and the type strain of A. gyllenbergii sp. nov. is NIPH 2150T (=RUH 422T=CCUG 51248T=CCM 7267T=1271T). PMID- 19126735 TI - Paenibacillus contaminans sp. nov., isolated from a contaminated laboratory plate. AB - A bacterial strain, designated CKOBP-6T, isolated from a contaminated laboratory plate, was characterized by using a polyphasic taxonomic approach. Chemotaxonomic and morphological properties of the strain were consistent with its classification in the genus Paenibacillus. Cells were Gram-variable, aerobic, sporulating, motile rods. 16S rRNA gene sequence analysis demonstrated that this novel isolate was unique, showing 94.6 % sequence similarity to Paenibacillus chitinolyticus NBRC 15660T and lower levels of similarity to Paenibacillus gansuensis KCTC 3950T (93.8 %) and Paenibacillus terrigena A35T (93.8 %). The novel isolate could be distinguished from the type strains of all of these species based on a range of phenotypic data. The major cellular phospholipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and one unknown phosphoglycolipid. The predominant isoprenologue was an unsaturated menaquinone with seven isoprene units (MK-7). The major fatty acids of strain CKOBP-6T were C(16 : 0) (12.0 %), anteiso-C(15 : 0) (52.0 %) and iso-C(16 : 0) (16.4 %). The G+C content of the genomic DNA was 51.2 mol%. It is evident from the genotypic and phenotypic data that strain CKOBP-6T should be classified as a representative of a novel species of the genus Paenibacillus, for which the name Paenibacillus contaminans sp. nov. is proposed. The type strain is CKOBP-6T (=BCRC 17728T=LMG 24216T). PMID- 19126736 TI - Paenibacillus siamensis sp. nov., Paenibacillus septentrionalis sp. nov. and Paenibacillus montaniterrae sp. nov., xylanase-producing bacteria from Thai soils. AB - Three strains of xylanase-producing bacteria, S5-3(T), X13-1(T) and MXC2-2(T), isolated from soils in Thailand, were characterized taxonomically based on their phenotypic and chemotaxonomic characteristics and 16S rRNA gene sequence comparisons. They were Gram-positive, facultatively anaerobic, spore-forming, rod shaped bacteria. They contained meso-diaminopimelic acid in the cell-wall peptidoglycan. The DNA G+C contents were respectively 45.8, 47.3 and 48.8 mol%. The predominant isoprenoid quinone was MK-7, and anteiso-C(15 : 0) and C(16 : 0) were the dominant cellular fatty acids. Phylogenetic analyses based on 16S rRNA gene sequence comparison showed that they were affiliated to the genus Paenibacillus. Strains S5-3(T), X13-1(T) and MXC2-2(T) were closely related to Paenibacillus granivorans A30(T) and Paenibacillus agaridevorans DSM 1355(T) (94.7-98.6 % 16S rRNA gene sequence similarity). The similarity between the three novel strains ranged from 96.3 to 98.4 %. DNA-DNA relatedness, DNA G+C contents and some phenotypic characteristics could clearly distinguish the strains from each other and from related Paenibacillus species. Therefore, strains S5-3(T), X13-1(T) and MXC2-2(T) represent novel species of the genus Paenibacillus, for which the names Paenibacillus siamensis sp. nov. (type strain S5-3(T) =KCTC 13038(T) =PCU 279(T) =TISTR 1831(T)), Paenibacillus septentrionalis sp. nov. (type strain X13-1(T) =KCTC 13039(T) =PCU 280(T) =TISTR 1830(T)) and Paenibacillus montaniterrae sp. nov. (type strain MXC2-2(T) =KCTC 13036(T) =PCU 281(T) =TISTR 1836(T)) are proposed. PMID- 19126737 TI - Nitratireductor basaltis sp. nov., isolated from black beach sand. AB - A Gram-negative, coccoid- or rod-shaped bacterium was isolated from black sand collected from Soesoggak beach, Jeju Island, Korea. The isolate, designated J3T, grew at 15-45 degrees C, at pH 5.5-10.0 and in 0-8 % NaCl. It was oxidase- and catalase-positive. Strain J3T reduced nitrate to nitrite, but did not reduce nitrite to nitrogen gas. Phylogenetic analysis of the 16S rRNA gene sequence showed that strain J3T was closely related to Nitratireductor aquibiodomus NL21T and belonged to the genus Nitratireductor. Major cellular fatty acids were C(18 : 1)omega7c (82.0 %), C(19 : 0)omega8c cyclo (4.3 %) and C(18 : 0) (4.0 %), a profile that is typical of members of the genus Nitratireductor and distinct from those of other genera in the family Phyllobacteriaceae. Differences in physiological characteristics and fatty acid profiles, as well as low DNA-DNA hybridization values, further established that strain J3T was distinct from N. aquibiodomus NL21T. Thus, strain J3T (=KCTC 22119T =JCM 14935T) should be classified as the type strain of a novel species in the genus Nitratireductor, for which the name Nitratireductor basaltis sp. nov. is proposed. PMID- 19126738 TI - Pseudomonas tuomuerensis sp. nov., isolated from a bird's nest. AB - Strain 78-123T was isolated from a sample of a bird's nest situated on the bank of Qiongtailan River in the region of Tuomuer Peak of Tianshan Mountain in the Xin-jiang Uygur Autonomous Region in north-western China. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain 78-123T was related to members of the genus Pseudomonas. 16S rRNA gene sequence similarity between strain 78-123T and Pseudomonas mendocina ATCC 25411T, Pseudomonas pseudoalcaligenes JCM 5968T and Pseudomonas alcaliphila AL15-21T was 97.1, 97.4 and 97.5 %, respectively. The major cellular fatty acids were C(16 : 0), C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, C(18 : 1)omega7c and C(12 : 0). The G+C content was 60.4 mol%. On the basis of the phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, the novel species Pseudomonas tuomuerensis sp. nov. is proposed, with the type strain 78-123T (=CGMCC 1.1365T =JCM 14085T). PMID- 19126739 TI - Deinococcus aquiradiocola sp. nov., isolated from a radioactive site in Japan. AB - A gamma- and UV-radiation-tolerant, pale-pink strain (TDMA-uv53T) was isolated from a freshwater sample collected at Misasa (Tottori, Japan), after exposure of the water sample to UV radiation. The cells stained Gram-positive and were non motile, rod-shaped and non-spore-forming. The DNA G+C content of the strain was 69.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain TDMA-uv53T belongs to the genus Deinococcus, the highest sequence similarities being found with Deinococcus claudionis PO-04-19-125T (96 %), D. altitudinis ME-04-01-32T (96 %), D. radiomollis PO-04-20-132T (95 %), D. deserti VCD115T (91.5 %), D. hopiensis KR-140T (91.0 %) and D. sonorensis KR-87T (91.0 %). Major fatty acids were iso-15 : 0, 15 : 1omega6c, 15 : 0, 16 : 0 and summed feature 3 (iso-15 : 0 2-OH and/or 16 : 1omega7c). MK-8 was the predominant respiratory quinone. Phylogenetic distinctiveness and unique phenotypic characteristics differentiated strain TDMA-uv53T from closely related Deinococcus species. The results of our polyphasic taxonomic analyses suggested that TDMA uv53T represents a novel Deinococcus species, for which the name Deinococcus aquiradiocola sp. nov. is proposed. The type strain is TDMA-uv53T (=JCM 14371(T) [corrected] =NBRC 102118T =CCUG 53612T). PMID- 19126740 TI - Robinsoniella peoriensis gen. nov., sp. nov., isolated from a swine-manure storage pit and a human clinical source. AB - A polyphasic taxonomic study was performed on six strains of an unknown Gram positive, non-motile, spore-forming, short oval to rod-shaped bacterium isolated from a swine-manure storage pit. In addition to these strains, an isolate deposited in the Culture Collection of the University of Goteborg (Sweden) was found to be biochemically related to the manure strains. The major end products of metabolism included acetate and succinate but not butyrate. Comparative 16S rRNA gene sequencing confirmed that all these isolates were closely related to each other and formed a hitherto unknown lineage within the clostridial rRNA XIVa cluster of organisms. On the basis of phylogenetic, biochemical and phenotypic evidence, it is proposed that the unknown bacterium represents a novel genus and species, for which the name Robinsoniella peoriensis gen. nov., sp. nov. is proposed. The type strain of Robinsoniella peoriensis is PPC31T (=CCUG 48729T =NRRL B-23985T). PMID- 19126741 TI - Novosphingobium panipatense sp. nov. and Novosphingobium mathurense sp. nov., from oil-contaminated soil. AB - In an ongoing attempt to analyse the diversity of culturable micro-organisms from oil-contaminated soil, two yellow-pigmented, Gram-negative, halophilic bacterial strains (SM16T and SM117T) were isolated. These two strains were characterized using a polyphasic approach. Strains SM16T and SM117T showed a range of phenotypic and chemotaxonomic properties that were consistent with those of members of the genus Novosphingobium. Both strains contained sphingoglycolipids, thus confirming that they belong to the Alphaproteobacteria. Furthermore, the polar lipid profile consisted of phosphatidylglycerol, phosphatidyldimethylethanolamine and phosphatidylethanolamine, with minor amounts of phosphatidyldimethylethanolamine, phosphatidylcholine and phosphatidylmonomethylethanolamine. Spermidine was the major polyamine in the cell wall, a characteristic feature of members of the genus Novosphingobium. Fatty acid analysis revealed the presence of C(18 : 1)omega7c, C(16 : 0) and C(14 : 0) 2-OH in both isolates, a characteristic feature of sphingomonads. 16S rRNA gene sequence similarities with the type strains of the most closely related species of the genus Novosphingobium (Novosphingobium pentaromativorans and Novosphingobium resinovorum) were less than 98.8 % for both strains. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains SM16T and SM117T fall in the clade represented by the genus Novosphingobium. Strains SM16T and SM117T shared 98.8 % similarity in their 16S rRNA gene sequences and their mean level of DNA-DNA relatedness was 8.5 %. Strains SM16T and SM117T differed from each other with respect to their morphological, physiological and chemotaxonomic properties. Thus, these results indicate that strains SM16T and SM117T belong to separate species of the genus Novosphingobium, for which the names Novosphingobium panipatense sp. nov. and Novosphingobium mathurense sp. nov. are proposed; the type strains are SM16T (=CCM 7472T =MTCC9019T) and SM117T (=CCM 7473T =MTCC9020T), respectively. PMID- 19126742 TI - Sphingobium ummariense sp. nov., a hexachlorocyclohexane (HCH)-degrading bacterium, isolated from HCH-contaminated soil. AB - A hexachlorocyclohexane (HCH)-degrading bacterial strain (RL-3T) was isolated from an HCH dump site located in the northern part of India. Resting cell assays and analytical GC studies confirmed the ability of strain RL-3T to degrade HCH isomers. Southern blot hybridization studies revealed the presence of lin genes, which are involved in the HCH degradation pathway, in this bacterium. The 16S rRNA gene sequence of strain RL-3T showed that it was most closely related to Sphingobium cloacae JCM 10874T (97.3 %) and Sphingobium fuliginis MTCC 7295T (96.4 %). Phylogenetic analysis based on 16S rRNA gene sequences placed strain RL 3T between S. cloacae JCM 10874T and S. fuliginis MTCC 7295T. The DNA G+C content of strain RL-3T was 62 mol%. The DNA-DNA relatedness values of strain RL-3T with S. cloacae JCM 10874T and S. fuliginis CCM 7327T were 8.65 and 7.47 %, respectively. SGL1 was the major sphingolipid and spermidine was the major polyamine in strain RL-3T. The major fatty acids in strain RL-3T were C(18 : 1)omega7c (56.6 %), C(16 : 0) (14 %) and C(14 : 0) 2-OH (7.4 %). Ubiquinone Q-10 was the major respiratory quinone. Phylogenetic distinctiveness, DNA-DNA relatedness values, biochemical and physiological characterization and unique phenotypic characteristics suggest that strain RL-3T represents a novel species of the genus Sphingobium, for which the name Sphingobium ummariense sp. nov. is proposed. The type strain is RL-3T (=MTCC 8599T=CCM 7431T). PMID- 19126743 TI - Phylogenetic placement of the Cyrtolophosididae Stokes, 1888 (Ciliophora; Colpodea) and neotypification of Aristerostoma marinum Kahl, 1931. AB - The ciliate family Cyrtolophosididae Stokes, 1888 contains species that are poorly known from both the morphological and molecular perspectives. To further our understanding of this family, one species, Aristerostoma marinum Kahl, 1931, was redescribed. Cells in our population had a mean in vivo size of 15x8 microm. There were six rows of somatic kineties, as well as six dorsal kinetids belonging to sparsely ciliated somatic kineties. The oral apparatus comprised a bipartite paroral membrane and four adoral organelles. The optimal ecological tolerances for pH and O2 matched those of the environment in which the specimens were collected, but were different for salinity and temperature. To further test the phylogenetic placement of the family Cyrtolophosididae with increased taxon sampling, the small subunit rDNA of three morphospecies was characterized: A. marinum, Aristerostoma sp. ATCC 50986 and Pseudocyrtolophopsis alpestris. Unconstrained and constrained molecular analyses supported the non-monophyly of the order Cyrtolophosidida. The family Cyrtolophosididae fell out separately from the rest of its order. Haplotypes from previous environmental studies were also placed in a phylogenetic context within the class Colpodea. PMID- 19126744 TI - Cryptococcus tepidarius sp. nov., a thermotolerant yeast species isolated from a stream from a hot-spring area in Japan. AB - Anamorphic basidiomycetous yeast strains M9962T and M9963 were isolated from water samples collected from a small stream in Ohwakudani, Hakone, a hot-spring area in Japan. These belonged to a single species and were phylogenetically closely related to Bullera lagerstroemiae, although the ability to form ballistoconidia was not observed. Based on sequence analyses of the D1/D2 domain of the LSU rDNA and ITS regions and differences in G+C content, the name Cryptococcus tepidarius sp. nov. Takashima, Sugita, Toriumi et Nakase (Trichosporonales, Tremellomycetes, Basiomycota) is proposed for these isolates, with strain M9962T (=SP-5T =JCM 11965T =CBS 9427T) as the type strain. The strains grew in YM broth at 47 degrees C and in YM broth with the pH adjusted to 1.5 by HCl, indicating that the species is thermotolerant and acid-tolerant. PMID- 19126745 TI - Candida tumulicola sp. nov. and Candida takamatsuzukensis sp. nov., novel yeast species assignable to the Candida membranifaciens clade, isolated from the stone chamber of the Takamatsu-zuka tumulus. AB - During a survey of the mycobiota in the stone chamber of the Takamatsu-zuka tumulus in the village of Asuka, Nara Prefecture, Japan, we isolated 19 yeast strains assigned to the genus Candida from various samples, taken mainly from mouldy spots where the colour of the murals had changed to black, white or another tone, and from viscous gels (biofilms) on plaster walls. The 26S rDNA D1/D2 domain sequence-based phylogeny clearly indicates two groups of isolates. Polyphasic characterization, including morphological, physiological and chemotaxonomic characteristics, and sequence analysis of the 26S rDNA D1/D2 domain and ITS regions suggest that each group is assignable to one of two novel species within the Candida membranifaciens clade. Proposed herein are the names Candida tumulicola sp. nov. (originally T6517-9-5T; holotype JCM 15403T; isotypes CBS 10917T, NBRC 104392T) and Candida takamatsuzukensis sp. nov. (originally T4922-1-1T; holotype JCM 15410T; isotypes CBS 10916T, NBRC 104391T). The 26S rDNA D1/D2 domain sequence divergence indicates that C. tumulicola differs from Candida friedrichii NBRC 10277T, the type strain of the nearest species, in 15 nucleotides (3 %), whereas C. takamatsuzukensis differs from Candida insectorum NBRC 10283T and Pichia mexicana NBRC 10544T, the type strains of the nearest species, in 20 nucleotides (4 %). Both novel species are also clearly distinguishable from the species closest to them by various physiological characteristics. PMID- 19126751 TI - Heat shock response protects human peritoneal mesothelial cells from dialysate induced oxidative stress and mitochondrial injury. AB - BACKGROUND: Chronic peritoneal dialysis (PD) is one of the major therapies for uremic patients. However, the peritoneal mesothelial cells (PMCs) are subject to the injury by bioincompatible dialysates. The aim of this study is to investigate the protective roles and mechanisms of heat shock response in PMCs. METHODS: Primary cultured human PMCs (HPMCs) were subjected to commercial peritoneal dialysates. The cell viability was assayed by MTT test and Annexin V assay. The expression of HSPs was detected by Western blots analysis. Intracellular hydrogen peroxide and superoxide anion were detected using H(2)DCFDA and dHE probe, respectively, with flow cytometry. The mitochondrial membrane potential (DeltaPsim) of HPMCs was evaluated using JC1 probe with flow-cytometry. RESULTS: Exposure of HPMCs to 1.5%, 2.5%, and 4.25% dextrose, and 7.5% icodextrin dialysates, respectively, for 60 min resulted in significantly accumulation of intracellular reactive oxygen species (ROS), DeltaPsim loss, and cell death in HPMCs. Amino acid dialysates exhibited no significant cytotoxicity. Adjusting the acidity in 1.5% dextrose and icodextrin dialysate significantly attenuated the dialysate-induced ROS generation and cell death in HPMCs. Heat pretreatment (41 degrees C, 30 minutes), which induced HSP 27 and 72 syntheses, significantly attenuated the dialysate-induced intracellular ROS accumulation, Dym loss, and cell death in HPMCs. CONCLUSIONS: In conclusion, the acidic bioincompatible dialysates induce oxidative stress, DeltaPsim loss, and subsequent cell death in HPMCs. Amino acid dialysates is more biocompatible than glucose and icodextrin dialysates to HPMCs. Heat shock response protects HPMCs from the bioincompatible dialysates-induced cellular damage. PMID- 19126752 TI - Congenic strains provide evidence that four mapped loci in chromosomes 2, 4, and 16 influence hypertension in the SHR. AB - To dissect the genetic architecture controlling blood pressure (BP) regulation in the spontaneously hypertensive rat (SHR) we derived congenic rat strains for four previously mapped BP quantitative trait loci (QTLs) in chromosomes 2, 4, and 16. Target chromosomal regions from the Brown Norway rat (BN) averaging 13-29 cM were introgressed by marker-assisted breeding onto the SHR genome in 12 or 13 generations. Under normal salt intake, QTLs on chromosomes 2a, 2c, and 4 were associated with significant changes in systolic BP (13, 20, and 15 mmHg, respectively), whereas the QTL on chromosome 16 had no measurable effect. On high salt intake (1% NaCl in drinking water for 2 wk), the chromosome 16 QTL had a marked impact on SBP, as did the QTLs on chromosome 2a and 2c (18, 17, and 19 mmHg, respectively), but not the QTL on chromosome 4. Thus these four QTLs affected BP phenotypes differently: 1) in the presence of high salt intake (chromosome 16), 2) only associated with normal salt intake (chromosome 4), and 3) regardless of salt intake (chromosome 2c and 2a). Moreover, salt sensitivity was abrogated in congenics SHR.BN2a and SHR.BN16. Finally, we provide evidence for the influence of genetic background on the expression of the mapped QTLs individually or as a group. Collectively, these data reveal previously unsuspected nuances of the physiological roles of each of the four mapped BP QTLs in the SHR under basal and/or salt loading conditions unforeseen by the analysis of the F2 cross. PMID- 19126753 TI - Interpreting neonatal lethal phenotypes in mouse mutants: insights into gene function and human diseases. AB - The mouse represents the model of choice to study the biological function of mammalian genes through mutation of its genome. However, the biggest challenge of mouse geneticists remains the phenotypic analysis of mouse mutants. A survey of mouse mutant databases reveals a surprisingly high number of gene mutations leading to neonatal death. These genetically modified mouse mutants have been instrumental in elucidating gene function and have become important models of congenital human diseases. The main complication when phenotyping mutant mice dying during the neonatal period is the large spectrum of physiological systems whose defects can challenge neonatal survival. Here, we present a comprehensive review of gene mutations leading to neonatal lethality and discuss the impact of these mutations on the major physiological processes critical to mouse newborn survival: parturition, breathing, suckling, and homeostasis. Selected examples of mouse mutants are highlighted to illustrate how the precise identification of the timing and cause of death associated with these physiological processes allows for a more profound understanding of the underlying cellular and molecular defects. This review provides a guide for the analysis of neonatal lethal phenotypes in mutant mice that will be helpful for dissecting out the function of specific genes during mouse development. PMID- 19126754 TI - Positive and negative regulation of insulin signaling by reactive oxygen and nitrogen species. AB - Regulated production of reactive oxygen species (ROS)/reactive nitrogen species (RNS) adequately balanced by antioxidant systems is a prerequisite for the participation of these active substances in physiological processes, including insulin action. Yet, increasing evidence implicates ROS and RNS as negative regulators of insulin signaling, rendering them putative mediators in the development of insulin resistance, a common endocrine abnormality that accompanies obesity and is a risk factor of type 2 diabetes. This review deals with this dual, seemingly contradictory, function of ROS and RNS in regulating insulin action: the major processes for ROS and RNS generation and detoxification are presented, and a critical review of the evidence that they participate in the positive and negative regulation of insulin action is provided. The cellular and molecular mechanisms by which ROS and RNS are thought to participate in normal insulin action and in the induction of insulin resistance are then described. Finally, we explore the potential usefulness and the challenges in modulating the oxidant-antioxidant balance as a potentially promising, but currently disappointing, means of improving insulin action in insulin resistance-associated conditions, leading causes of human morbidity and mortality of our era. PMID- 19126757 TI - Role of bile acids and bile acid receptors in metabolic regulation. AB - The incidence of the metabolic syndrome has taken epidemic proportions in the past decades, contributing to an increased risk of cardiovascular disease and diabetes. The metabolic syndrome can be defined as a cluster of cardiovascular disease risk factors including visceral obesity, insulin resistance, dyslipidemia, increased blood pressure, and hypercoagulability. The farnesoid X receptor (FXR) belongs to the superfamily of ligand-activated nuclear receptor transcription factors. FXR is activated by bile acids, and FXR-deficient (FXR(-/ )) mice display elevated serum levels of triglycerides and high-density lipoprotein cholesterol, demonstrating a critical role of FXR in lipid metabolism. In an opposite manner, activation of FXR by bile acids (BAs) or nonsteroidal synthetic FXR agonists lowers plasma triglycerides by a mechanism that may involve the repression of hepatic SREBP-1c expression and/or the modulation of glucose-induced lipogenic genes. A cross-talk between BA and glucose metabolism was recently identified, implicating both FXR-dependent and FXR-independent pathways. The first indication for a potential role of FXR in diabetes came from the observation that hepatic FXR expression is reduced in animal models of diabetes. While FXR(-/-) mice display both impaired glucose tolerance and decreased insulin sensitivity, activation of FXR improves hyperglycemia and dyslipidemia in vivo in diabetic mice. Finally, a recent report also indicates that BA may regulate energy expenditure in a FXR-independent manner in mice, via activation of the G protein-coupled receptor TGR5. Taken together, these findings suggest that modulation of FXR activity and BA metabolism may open new attractive pharmacological approaches for the treatment of the metabolic syndrome and type 2 diabetes. PMID- 19126758 TI - Physiology of cell volume regulation in vertebrates. AB - The ability to control cell volume is pivotal for cell function. Cell volume perturbation elicits a wide array of signaling events, leading to protective (e.g., cytoskeletal rearrangement) and adaptive (e.g., altered expression of osmolyte transporters and heat shock proteins) measures and, in most cases, activation of volume regulatory osmolyte transport. After acute swelling, cell volume is regulated by the process of regulatory volume decrease (RVD), which involves the activation of KCl cotransport and of channels mediating K(+), Cl(-), and taurine efflux. Conversely, after acute shrinkage, cell volume is regulated by the process of regulatory volume increase (RVI), which is mediated primarily by Na(+)/H(+) exchange, Na(+)-K(+)-2Cl(-) cotransport, and Na(+) channels. Here, we review in detail the current knowledge regarding the molecular identity of these transport pathways and their regulation by, e.g., membrane deformation, ionic strength, Ca(2+), protein kinases and phosphatases, cytoskeletal elements, GTP binding proteins, lipid mediators, and reactive oxygen species, upon changes in cell volume. We also discuss the nature of the upstream elements in volume sensing in vertebrate organisms. Importantly, cell volume impacts on a wide array of physiological processes, including transepithelial transport; cell migration, proliferation, and death; and changes in cell volume function as specific signals regulating these processes. A discussion of this issue concludes the review. PMID- 19126756 TI - Transcription factors in long-term memory and synaptic plasticity. AB - Transcription is a molecular requisite for long-term synaptic plasticity and long term memory formation. Thus, in the last several years, one main interest of molecular neuroscience has been the identification of families of transcription factors that are involved in both of these processes. Transcription is a highly regulated process that involves the combined interaction and function of chromatin and many other proteins, some of which are essential for the basal process of transcription, while others control the selective activation or repression of specific genes. These regulated interactions ultimately allow a sophisticated response to multiple environmental conditions, as well as control of spatial and temporal differences in gene expression. Evidence based on correlative changes in expression, genetic mutations, and targeted molecular inhibition of gene expression have shed light on the function of transcription in both synaptic plasticity and memory formation. This review provides a brief overview of experimental work showing that several families of transcription factors, including CREB, C/EBP, Egr, AP-1, and Rel, have essential functions in both processes. The results of this work suggest that patterns of transcription regulation represent the molecular signatures of long-term synaptic changes and memory formation. PMID- 19126755 TI - Mammalian nicotinic acetylcholine receptors: from structure to function. AB - The classical studies of nicotine by Langley at the turn of the 20th century introduced the concept of a "receptive substance," from which the idea of a "receptor" came to light. Subsequent studies aided by the Torpedo electric organ, a rich source of muscle-type nicotinic receptors (nAChRs), and the discovery of alpha-bungarotoxin, a snake toxin that binds pseudo-irreversibly to the muscle nAChR, resulted in the muscle nAChR being the best characterized ligand-gated ion channel hitherto. With the advancement of functional and genetic studies in the late 1980s, the existence of nAChRs in the mammalian brain was confirmed and the realization that the numerous nAChR subtypes contribute to the psychoactive properties of nicotine and other drugs of abuse and to the neuropathology of various diseases, including Alzheimer's, Parkinson's, and schizophrenia, has since emerged. This review provides a comprehensive overview of these findings and the more recent revelations of the impact that the rich diversity in function and expression of this receptor family has on neuronal and nonneuronal cells throughout the body. Despite these numerous developments, our understanding of the contributions of specific neuronal nAChR subtypes to the many facets of physiology throughout the body remains in its infancy. PMID- 19126760 TI - Endocannabinoid-mediated control of synaptic transmission. AB - The discovery of cannabinoid receptors and subsequent identification of their endogenous ligands (endocannabinoids) in early 1990s have greatly accelerated research on cannabinoid actions in the brain. Then, the discovery in 2001 that endocannabinoids mediate retrograde synaptic signaling has opened up a new era for cannabinoid research and also established a new concept how diffusible messengers modulate synaptic efficacy and neural activity. The last 7 years have witnessed remarkable advances in our understanding of the endocannabinoid system. It is now well accepted that endocannabinoids are released from postsynaptic neurons, activate presynaptic cannabinoid CB(1) receptors, and cause transient and long-lasting reduction of neurotransmitter release. In this review, we aim to integrate our current understanding of functions of the endocannabinoid system, especially focusing on the control of synaptic transmission in the brain. We summarize recent electrophysiological studies carried out on synapses of various brain regions and discuss how synaptic transmission is regulated by endocannabinoid signaling. Then we refer to recent anatomical studies on subcellular distribution of the molecules involved in endocannabinoid signaling and discuss how these signaling molecules are arranged around synapses. In addition, we make a brief overview of studies on cannabinoid receptors and their intracellular signaling, biochemical studies on endocannabinoid metabolism, and behavioral studies on the roles of the endocannabinoid system in various aspects of neural functions. PMID- 19126761 TI - Cumulative energy imbalance in the pediatric intensive care unit: role of targeted indirect calorimetry. AB - INTRODUCTION: Failure to accurately estimate energy requirements may result in underfeeding or overfeeding. In this study, a dedicated multidisciplinary nutrition team measured energy expenditure in critically ill children. METHODS: Steady-state indirect calorimetry was used to obtain measured resting energy expenditure, which was compared with equation-estimated energy expenditure and the total energy intake for each subject. The children's metabolic status was examined in relation to standard clinical characteristics. RESULTS: Sixteen measurements were performed in 14 patients admitted to the multidisciplinary pediatric intensive care unit over a period of 12 months. Mean age of subjects in this cohort was 11.2 years (range 1.6 months to 32 years) and included 7 males and 7 postoperative patients. Altered metabolism was detected in 13 of 14 subjects and in 15 of 16 (94%) measurements. There was no correlation between the metabolic status of subjects and their clinical characteristics. Average daily energy balance was 200 kcal/d (range -518 to +859 kcal/d). Agreement between measured resting energy expenditure and equation-estimated energy expenditure was poor, with mean bias of 72.3 +/- 446 kcal/d (limits of agreement -801.9 to + 946.5 kcal/d). CONCLUSIONS: A disparity was observed between equation-estimated energy expenditure, measured resting energy expenditure, and total energy intake, with a high incidence of underfeeding or overfeeding. A wide range of metabolic alterations were recorded, which could not be accurately predicted using standard clinical characteristics. Targeted indirect calorimetry on high-risk patients selected by a dedicated nutrition team may prevent cumulative excesses and deficits in energy balance. PMID- 19126763 TI - Willing, ready, and able? How we must exercise leadership for needed change in dental education. AB - For over twenty-five years, dental education has had the benefit of environmental analyses and institutional planning for change. Strong programs for leadership development have emerged to give direction to these efforts. Leading and thriving, not merely surviving, are universal aspirations, yet we remain vexed by finances, structures, and traditions. This article takes a look at change and examines the difference between technical frameworks for leadership and adaptive leadership. Leadership for change is viewed as an activity, not as a position of formal authority. The skills necessary to address the beliefs, attitudes, and culture that place limiting boundaries on adaptive leadership are described. Using the work of Heifetz and Linsky, the relationship between authority and adaptive leadership is defined. Resistance to change is presented as reaction to loss, which needs to be addressed in a fundamental way, through leadership activity and engagement. If change and innovation are to be sustained, leadership must be less accidental, less technical, and more adaptive. PMID- 19126762 TI - Starvation-induced proximal gut mucosal atrophy diminished with aging. AB - BACKGROUND: Starvation induces small bowel atrophy with increased intestinal epithelial apoptosis and decreased proliferation. The authors examined these parameters after starvation in aged animals. METHODS: Sixty-four 6-week-old and 26-month-old C57BL/6 mice were randomly assigned to either an ad libitum fed or fasted group. The small bowel was harvested at 12, 48, and 72 hours following starvation. Proximal gut mucosal height was measured and epithelial cells counted. Apoptosis was identified by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining. Proliferation was determined by immunohistochemical staining for proliferating cell nuclear antigen. Comparison of fed vs fasted and adult vs old groups was done by one-way ANOVA with Tukey's test and unpaired Student's t test. Significance was accepted at P < .05. RESULTS: Aged mice had higher proximal gut weights, mucosal heights, and cell numbers at baseline compared with the adult group (P < .05). The rate of apoptosis was lower in the aged (P < .05), but proliferation was not different between groups before starvation. After starvation, proximal gut wet weight decreased only in adult mice (P < .05). Gut mucosal height and mucosal cell number decreased more in adult than in aged mice (P < .05). This was related to decreased proliferation only in the adult group (P < .05). The fold of epithelial apoptosis that increased was higher in the aged group than in the adult group after starvation (P < .05). CONCLUSIONS: Gut mucosal kinetics change with age and have lower rates of apoptosis and greater mucosal mass; the character of starvation-induced atrophy is diminished with aging. PMID- 19126759 TI - Cardiovascular actions of neurotrophins. AB - Neurotrophins were christened in consideration of their actions on the nervous system and, for a long time, they were the exclusive interest of neuroscientists. However, more recently, this family of proteins has been shown to possess essential cardiovascular functions. During cardiovascular development, neurotrophins and their receptors are essential factors in the formation of the heart and critical regulator of vascular development. Postnatally, neurotrophins control the survival of endothelial cells, vascular smooth muscle cells, and cardiomyocytes and regulate angiogenesis and vasculogenesis, by autocrine and paracrine mechanisms. Recent studies suggest the capacity of neurotrophins, via their tropomyosin-kinase receptors, to promote therapeutic neovascularization in animal models of hindlimb ischemia. Conversely, the neurotrophin low-affinity p75(NTR) receptor induces apoptosis of endothelial cells and vascular smooth muscle cells and impairs angiogenesis. Finally, nerve growth factor looks particularly promising in treating microvascular complications of diabetes or reducing cardiomyocyte apoptosis in the infarcted heart. These seminal discoveries have fuelled basic and translational research and thus opened a new field of investigation in cardiovascular medicine and therapeutics. Here, we review recent progress on the molecular signaling and roles played by neurotrophins in cardiovascular development, function, and pathology, and we discuss therapeutic potential of strategies based on neurotrophin manipulation. PMID- 19126764 TI - Dental student assessment toolbox. PMID- 19126765 TI - Why teach? One dental student's views from both sides of the lectern. PMID- 19126766 TI - Periodontal-systemic disease education in U.S. and Canadian dental schools. AB - Research has proliferated in recent years regarding the relationship of oral disease to systemic conditions. Specifically, periodontal disease has been studied as a potential risk factor for multiple conditions such as cardiovascular disease (CVD) and adverse pregnancy outcomes, while other research focuses on exposures or behaviors associated with oral disease. However, few articles have been published reporting how this information is integrated into schools of dentistry, both in the classroom and clinical curriculum. For our study, a thirty three-item survey and cover letter were electronically mailed to academic deans at sixty-five accredited dental schools in the United States and Canada in the fall of 2007. The response rate was 77 percent. According to the responses to this survey, the primary topics covered in the didactic curriculum regarding periodontal oral-systemic disease are aging, CVD, diabetes, and tobacco use. Eighty-eight percent of the respondents reported that their students are knowledgeable about the role of inflammation and its impact on oral-systemic conditions. Forty-eight percent of the respondents said they provide formal training for their students in how to discuss or communicate aspects of periodontal oral-systemic disease with patients. Only seven schools reported teaching didactic content to dental students intermixed with other health professions students, and only two schools reported conducting joint projects. Only 9 percent of the respondents said they think nurses and physicians are knowledgeable about oral-systemic disease. The findings indicate that dental schools are confident about the knowledge of their students regarding oral systemic content. However, much work is needed to educate dental students to work in a collaborative fashion with other health care providers to co-manage patients at risk for oral-systemic conditions. PMID- 19126767 TI - Finding a method to analyze qualitative data: using a study of conceptual learning. AB - There is increasing awareness in the health sciences of the potential of qualitative research to address questions that quantitative research cannot satisfactorily answer. While a growing number of studies in health sciences and health sciences education discuss the value of such research or describe the methodology and data collection processes, few detail how analysis was carried out. Reliability and validity of findings from qualitative research depend on the quality of data management, retrieval, and interpretation or identification of meaning. The robustness of data analysis is therefore an important factor in the rigor of qualitative research. This article uses a study of dental students' conceptual learning to illustrate strategies that ensure rigor in qualitative analysis. Factors that informed the decisions regarding analysis are discussed in detail. The use of both grounded theory and literature is discussed. The role that deductive and inductive reasoning played in the analysis is outlined. A brief section illustrates the kinds of conclusions that can be made about conceptual learning when qualitative data are rigorously analyzed. Finally, potential shortcomings in the study and alternatives or additional mechanisms for ensuring validity and reliability of analysis are discussed. PMID- 19126768 TI - Lessons from students in a critical thinking course: a case for the third pedagogy. AB - There have been many calls for strengthening the level of critical thinking among dental students and faculty members, but few analyses of how such curricular experiences actually affect them. This report provides a rich, multifaceted description of eight years' experience teaching a course in critical thinking. Among the data analyzed were a) course materials and learning experiences, b) student products demonstrating critical thinking, c) tests of subject matter knowledge, d) content analysis of what students and faculty members find problematic, e) a test of critical thinking aptitude, f) two personality inventories, g) measures of didactic and clinical performance, and h) student comments and evaluation of instruction. These data support the view that critical thinking involves more than knowledge and application of skills relative to science and research that can be taught using traditional didactic methods. Personality factors emerged as predictive of both critical thinking and clinical performance, and students' approaches to learning appeared to be influenced by their practical considerations of what is needed to practice, rather than by the logic of good science. This analysis argues for designing curricula on the full concept of competency (knowledge, skills, and values needed to function as a dental practitioner) and for exploring a third pedagogy of reflective practice to supplement the traditional ones of didactic and skill teaching in dental education. PMID- 19126769 TI - The development and implementation of an online applied biochemistry bridge course for a dental hygiene curriculum. AB - This article describes a curricular change project designed to improve instruction in biochemistry. After years of unsatisfactory outcomes from a dental hygiene biochemistry course, a decision was made to change the traditional lecture-based course to an online format. Using online technology and principles of educational pedagogy, a course was developed that fosters application of biomaterials principles to dental hygiene practice and provides a bridge between prerequisite chemistry coursework and biochemistry in a health professions program. Members of the dental hygiene graduating Classes of 2007 and 2008 participated in the revised course. The outcome measures used to assess the effectiveness of the revised course were student end-of-semester course evaluations, graduating senior survey results, student course performance, and National Board examination performance. While the results are based on only two classes, the positive outcomes suggest that the revision was a worthwhile endeavor. The use of technology in teaching holds the potential for solving many of the curriculum and instruction issues currently under discussion: overcrowding of the curriculum, lack of active learning methods, and basic sciences taught in isolation from the rest of the curriculum. It is hoped that the results of this change will be helpful to other faculty members seeking curricular change and innovation. PMID- 19126770 TI - Factors that influence dental students' attitudes about older adults. AB - Our study considered dental students' general attitudes towards older persons using the Aging Semantic Differential. The influence of age, gender, cohort, education, and academic exposure on general attitudes towards older adults was evaluated using a total of 328 dental students across all four years of academic standing. Students were assessed in the fall and spring semesters. The results showed differential responding on the four subscales, with slight positive ratings on the autonomy, acceptability, and integrity subscales and a slight negative rating for instrumentality. Females expressed more negative attitudes than their male counterparts, with no age differences. There was also no significant impact from a specific, didactic educational component offered to the fourth-year students. However, the fourth-year students were the only group to show positive changes across the full academic year. The results suggest that general attitudes can be changed, but didactic (classroom) forms of education alone are insufficient to meaningfully modify students' perceptions of the elderly. Exposure to older adults in a clinical setting appears to be a critical element, as the fourth-year students received much greater exposure to older patients and more intensified interface with their mentors. PMID- 19126771 TI - Lesbian, gay, bisexual, and transgender (LGBT) issues in dental school environments: dental student leaders' perceptions. AB - The objectives of the study reported in this article were to assess dental student leaders' perceptions of educational efforts concerning lesbian, gay, bisexual, and transgender (LGBT) topics and the cultural climate concerning LGBT issues in dental schools in the United States and Canada. In addition, the perceptions of student leaders who self-identified as belonging to the LGBT community and of students with a heterosexual orientation were compared. Data were collected from 113 dental student leaders from twenty-seven dental schools in the United States and three in Canada. Fifty student leaders were females, and sixty-two were males. Only 13.3 percent of the respondents agreed that their dental education prepared them well to treat patients from LGBT backgrounds. The more the student leaders believed that their university has an honest interest in diversity, the better they felt prepared by their dental school program to treat patients from LGBT backgrounds (r=.327; p<.001). The better they felt prepared, the more they perceived the clinic environment as sensitive and affirming for patients with different sexual orientations (r=.464; p<.001). The more they reported that dental schools' administrations create a positive environment for students with LGBT orientations, the more they agreed that persons can feel comfortable regardless of their sexual orientation (r=.585; p<.001). In conclusion, the findings indicate that dental school administrators play an important role in ensuring that future care providers are well prepared to treat patients from LGBT backgrounds and that staff, faculty, students, and patients from these backgrounds are not discriminated against. PMID- 19126772 TI - Dental students' and faculty members' perceptions of incivility in the classroom. AB - The purpose of this study was to compare dental students' and faculty members' perceptions of classroom incivility and to determine how many students willingly engage in uncivil behavior in the classroom. A web-based survey research design was used to obtain information regarding uncivil classroom behaviors from dental students and full- and part-time dental faculty members at a midwestern U.S. dental school. Responses were received from sixty-eight dental faculty members and 127 third- and fourth-year students. The perceptions of faculty members about what constitutes uncivil classroom behavior differed from those of students. In fact, of the eighteen questions asked of both groups, statistical differences were found for eleven (61 percent). PMID- 19126773 TI - Student-to-student local anesthesia injections in dental education: moral, ethical, and legal issues. AB - This article reports the findings of a survey-based study conducted to determine U.S. dental schools' institutional protocols regarding the practice of students' administering local anesthetic injections to fellow students as part of their process of learning this skill. The majority of schools ask students to practice local anesthetic injections on each other without obtaining informed consent. PMID- 19126774 TI - Should preclinical typodonts be disinfected prior to grading? AB - This is a report of a unique finding in a preclinical laboratory that may be a potential dental school health hazard. Visual inspection (conducted in April 2008 by a preclinical crown and bridge course coordinator) of typodonts used by second year students at the University of Mississippi School of Dentistry found that fourteen out of thirty-nine had black spots on the undersurface of the cheek shroud and/or plastic gingiva. The spots were cultured by the Medical Center's Department of Microbiology and described only as being mold/fungus typical of that which frequently grows in warm, moist, southern environments. Although indoor molds are common, about 5 percent of the general population will develop some type of mild allergic airway problem from molds over their lifetime. Mold on typodonts is unsightly, indicates failure of students to recognize the value of cleanliness in the dental environment, and may be a potential health hazard for some individuals. Cleaning and drying procedures for typodonts were implemented. The transfer of items between students and instructors during preclinical courses provides many opportunities for the spread of potentially harmful microorganisms/viruses. As a minimal level of personal protection, it is suggested that instructors wear disposable gloves and face masks and exercise hand washing between handling student instruments and typodonts. This problem has not been previously mentioned in the literature and merits further investigation/discussion. PMID- 19126776 TI - MEIS1 intronic risk haplotype associated with restless legs syndrome affects its mRNA and protein expression levels. AB - Restless legs syndrome (RLS) is a common neurological disorder characterized by an irresistible urge to move the legs at night, which is often accompanied by unpleasant sensations. A recent genomewide association study identified an association between RLS and intronic markers from the MEIS1 gene. Comparative genomic analysis indicates that MEIS1 is the only gene encompassed in this evolutionarily conserved chromosomal segment, i.e. a conservation synteny block, from mammals to fish. We carried out a series of experiments to delineate the role of MEIS1 in RLS pathogenesis and the underlying genetic mechanism. We sequenced all 13 MEIS1 exons and their splice junctions in 285 RLS probands with confirmed clinical diagnosis and did not identify any causative coding or exon intron junction mutations. We found no evidence of structural variation or disease-associated haplotype differential splicing. However, sequencing of conserved regions of MEIS1 introns 8 and 9 identified a novel single nucleotide polymorphism (C13B_2) significantly associated with RLS (allelic association, P = 1.81E-07). We detected a significant decrease in MEIS1 mRNA expression by quantitative real-time polymerase chain reaction in lymphoblastoid cell lines (LCLs) and brain tissues from RLS patients homozygous for the intronic RLS risk haplotype, compared with those homozygous for the non-risk haplotype. Finally, we found significantly decreased MEIS1 protein levels in the same batch of LCLs and brain tissues from the homozygous carriers of the risk haplotype, compared with the homozygous non-carriers. Therefore, these data suggest that reduced expression of the MEIS1 gene, possibly through intronic cis-regulatory element(s), predisposes to RLS. PMID- 19126778 TI - Dominant cataract formation in association with a vimentin assembly disrupting mutation. AB - Cataracts are characterized by an opacification of the eye lens, often caused by protein misfolding and aggregation. The intermediate filament protein vimentin, which is highly expressed in lens fiber cells and in mesenchymal tissues, is a main structural determinant in these cells forming a membrane-connected cytoskeleton. Additional functions of vimentin remain to be identified. Here, we demonstrate that a mutation in VIM causes a dominant, pulverulent cataract. We sequenced the complete human VIM gene in 90 individuals suffering from congenital cataract and found a G596A change in exon 1 in a single individual, causing the missense mutation E151K in coil 1B of vimentin. The mutant vimentin formed an aberrant vimentin cytoskeleton and increased the proteasome activity in transfected cells. Furthermore, this mutation causes a severe kinetic defect in vimentin assembly both in vitro and in vivo. Hence, in conjunction with available mouse and cell culture models, our results reveal for the first time an important functional role for vimentin in the maintenance of lens integrity. Finally, this invites novel therapy approaches for cataracts. PMID- 19126777 TI - Association of ESR1 gene tagging SNPs with breast cancer risk. AB - We have conducted a three-stage, comprehensive single nucleotide polymorphism (SNP)-tagging association study of ESR1 gene variants (SNPs) in more than 55,000 breast cancer cases and controls from studies within the Breast Cancer Association Consortium (BCAC). No large risks or highly significant associations were revealed. SNP rs3020314, tagging a region of ESR1 intron 4, is associated with an increase in breast cancer susceptibility with a dominant mode of action in European populations. Carriers of the c-allele have an odds ratio (OR) of 1.05 [95% Confidence Intervals (CI) 1.02-1.09] relative to t-allele homozygotes, P = 0.004. There is significant heterogeneity between studies, P = 0.002. The increased risk appears largely confined to oestrogen receptor-positive tumour risk. The region tagged by SNP rs3020314 contains sequence that is more highly conserved across mammalian species than the rest of intron 4, and it may subtly alter the ratio of two mRNA splice forms. PMID- 19126779 TI - Functional SNPs in the SCGB3A2 promoter are associated with susceptibility to Graves' disease. AB - Graves' disease (GD) is one of the most common human autoimmune diseases, and recent data estimated a prevalence of clinical hyperthyroidism of 0.25-1.09% in the population. Several reports have linked GD to the region 5q12-q33; and a locus between markers D5s436 and D5s434 was specifically linked to GD susceptibility in the Chinese population. In the present study, association analysis was performed using a large number of single-nucleotide polymorphisms (SNPs) at this locus in 2811 patients with GD recruited from different geographic regions of China. The strongest associations with GD in the combined Chinese Han cohorts were mapped to two SNPs in the promoter (pSNP) of SCGB3A2 [SNP76, rs1368408, P = 1.43 x 10(-6), odds ratio (OR) = 1.28 and SNP75, -623 - -622, P = 7.62 x 10(-5), OR = 1.32, respectively], a gene implicated in immune regulation. On the other hand, pSNP haplotypes composed of the SNP76 (rs1368408)+SNP74 (rs6882292) or SNP76+SNP75 (-623 approximately -622, AG/T) variants are correlated with high disease susceptibility (P = 0.0007, and P = 0.0192, respectively) in this combined Chinese Han cohort. Furthermore, these haplotypes were associated with reduced SCGB3A2 gene expression levels in human thyroid tissue, while functional analysis revealed a relatively low efficiency of SCGB3A2 promoters of the SNP76+SNP75 and SNP76+SNP74 haplotypes in driving gene expression. These results suggest that the SCGB3A2 gene may contribute to GD susceptibility. PMID- 19126780 TI - Quantitation of the contractile response mediated by two receptors: M2 and M3 muscarinic receptor-mediated contractions of human gastroesophageal smooth muscle. AB - Although muscarinic receptors are known to mediate tonic contraction of human gastrointestinal tract smooth muscle, the receptor subtypes that mediate the tonic contractions are not entirely clear. Whole human stomachs with attached esophagus were procured from organ transplant donors. Cholinergic contractile responses of clasp, sling, lower esophageal circular (LEC), midesophageal circular (MEC), and midesophageal longitudinal (MEL) muscle strips were determined. Sling fibers contracted greater than the other fibers. Total, M(2) and M(3) muscarinic receptor density was determined for each of these dissections by immunoprecipitation. M(2) receptor density is greatest in the sling fibers, followed by clasp, LEC, MEC, and then MEL, whereas M(3) density is greatest in LEC, followed by MEL, MEC, sling, and then clasp. The potency of subtype selective antagonists to inhibit bethanechol-induced contraction was calculated by Schild analysis to determine which muscarinic receptor subtypes contribute to contraction. The results suggest both M(2) and M(3) receptors mediate contraction in clasp and sling fibers. Thus, this type of analysis in which multiple receptors mediate the contractile response is inappropriate, and an analysis method relating dual occupation of M(2) and M(3) receptors to contraction is presented. Using this new method of analysis, it was found that the M(2) muscarinic receptor plays a greater role in mediating contraction of clasp and sling fibers than in LEC, MEC, and MEL muscles in which the M(3) receptor predominantly mediates contraction. PMID- 19126781 TI - The smoking cessation medication varenicline attenuates alcohol and nicotine interactions in the rat mesolimbic dopamine system. AB - Varenicline was recently approved as an aid for smoking cessation. Patients treated with varenicline have reported a concomitant reduction in their alcohol consumption. This compound has also been demonstrated to reduce alcohol seeking and consumption in alcohol high-preferring rats. Based on the extensive coabuse of nicotine and alcohol, the aim of the present study was to explore whether interactions among varenicline, nicotine, and ethanol in the brain reward system could indicate the use of varenicline also for alcohol dependence. Using the in vivo microdialysis method, we investigated the effects of systemic injections of varenicline on the extracellular accumbal dopamine levels in response to a systemic challenge of ethanol, nicotine, or the combination of nicotine and ethanol in the experimental rat. Acute systemic coadministration of varenicline and ethanol counteracted each others' respective enhancing effect on dopamine levels in the nucleus accumbens. However, after 5 days of varenicline pretreatment, acute combined varenicline and ethanol administration raised dopamine levels to the same extent as either drug alone. Furthermore, after varenicline pretreatment an acute injection of varenicline antagonized the dopamine stimulatory effect of acute nicotine as well as that of systemic coadministration of ethanol and nicotine. In contrast, a pronounced additive dopamine increase was observed when nicotine and ethanol were coadministered in vehicle-pretreated rats. The antismoking agent varenicline exhibits properties with respect to its interaction with ethanol and nicotine in the brain reward system that may be beneficial for treating patients with alcohol dependence with (and possibly also without) concomitant nicotine dependence. PMID- 19126782 TI - Genetic association of the activin A receptor gene (ACVR2A) and pre-eclampsia. AB - Pre-eclampsia is a common serious disorder of human pregnancy, which is associated with significant maternal and perinatal morbidity and mortality. The suspected aetiology of pre-eclampsia is complex, with susceptibility being attributable to multiple environmental factors and a large genetic component. Recently, we reported significant linkage to chromosome 2q22 in 34 Australian/New Zealand (Aust/NZ) pre-eclampsia/eclampsia families, and activin A receptor type IIA (ACVR2A) was identified as a strong positional candidate gene at this locus. In an attempt to identify the putative risk variants, we have now comprehensively re-sequenced the entire coding region of the ACVR2A gene and the conserved non coding sequences in a subset of 16 individuals from these families. We identified 45 single nucleotide polymorphisms (SNPs), with 9 being novel. These SNPs were genotyped in our total family sample of 480 individuals from 74 Aust/NZ pre eclampsia families (including the original 34 genome-scanned families). Our best associations between ACVR2A polymorphisms and pre-eclampsia were for rs10497025 (P = 0.025), rs13430086 (P = 0.010) and three novel SNPs: LF004, LF013 and LF020 (all with P = 0.018). After correction for multiple hypothesis testing, none of these associations reached significance (P > 0.05). Based on these data, it remains unclear what role, if any, ACVR2A polymorphisms play in pre-eclampsia risk, at least in these Australian families. However, it would be premature to rule out this gene as significant associations between ACVR2A SNPs and pre eclampsia have recently been reported in a large Norwegian (HUNT) population sample. PMID- 19126783 TI - Shotgun lipidomics of neutral lipids as an enabling technology for elucidation of lipid-related diseases. AB - Neutral lipids fulfill multiple specialized roles in cellular function. These roles include energy storage and utilization, the synthesis of complex lipids in cellular membranes, lipid second messengers for cellular signaling, and the modulation of membrane molecular dynamics. We have developed a novel mass spectrometric technology, now termed shotgun lipidomics, that can identify the types and amounts of thousands of lipids directly from extracts of biological samples. Shotgun lipidomics is well suited for the identification and measurement of the types and amounts of neutral lipid classes and individual molecular species through the use of multidimensional mass spectrometry. This review summarizes the basic principles underlying the use of shotgun lipidomics for the direct measurement of neutral lipids from extracts of biological tissues or fluids. Through exploiting the high information content inherent in shotgun lipidomics, this technology promises to greatly facilitate advances in our understanding of alterations in neutral lipid metabolism in health and disease. PMID- 19126784 TI - 20-Hydroxyecdysone decreases weight and hyperglycemia in a diet-induced obesity mice model. AB - The steroid hormone 20-hydroxyecdysone (20HE) is an essential signaling molecule that modulates molting response in insects and may function as a putative anabolic factor in vertebrate animals, although no mammalian 20HE receptor has been identified. Here we show that in H4IIE cell culture, 20HE treatment decreased expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6 phosphatase (G6Pase), reduced glucose production, and induced Akt2 phosphorylation sensitive to the phosphoinositide-3 kinase pathway-specific inhibitor LY-294002. Daily oral administration of 20HE (10 mg/kg for 13 wk) ameliorated obesity and insulin resistance in C57BL/6J mice fed a high-fat diet and produced a significant decrease of body weight gain and body fat mass compared with nontreated animals as demonstrated by dual-energy X-ray absorptiometry analysis. In addition, plasma insulin levels and glucose tolerance were significantly lowered by 20HE treatment. These changes were accompanied by the reduced hepatic expression of PEPCK and G6Pase and increased adiponectin production by visceral fat tissue. These studies demonstrate the anti-obesity and anti-diabetic effects of 20HE and begin to elucidate its putative cellular targets both in vitro and in vivo. PMID- 19126785 TI - Intracellular trafficking of the human oxytocin receptor: evidence of receptor recycling via a Rab4/Rab5 "short cycle". AB - As in the case of most G protein-coupled receptors, agonist stimulation of human oxytocin receptors (OTRs) leads to desensitization and internalization; however, little is known about the subsequent intracellular OTR trafficking, which is crucial for reestablishing agonist responsiveness. We examined receptor resensitization by first using HEK293T cells stably expressing human OTRs. Upon agonist activation, the receptors were almost completely sequestered inside intracellular compartments that were not labeled by lysosomal markers, thus indicating that the internalized receptors were not sorted to these degrading organelles. Binding and fluorescence assays showed that almost 85% of the receptors had returned to the cell surface after 4 h, by which time cell responsiveness to the agonist was also completely restored, as shown by measuring phospholipase C activation. Similar results were also obtained in the presence of cycloheximide, thus indicating that receptor recycling and not de novo receptor synthesis was responsible for the resensitization. Notably, very similar internalization and recycling kinetics were observed in endogenous OTRs expressed on myometrial cells. We also investigated the role of beta-arrestin2 in OTR recycling as these receptors have been previously classified as slowly or nonrecycling receptors on the basis of their stable association with this interacting protein. Our data suggest that the stable OTR/beta-arrestin2 interaction plays an important role in determining the rate of recycling of human OTRs, but does not determine the fate of endocytosed receptors. Subsequent investigations of receptor recycling pathways showed that OTRs localize in vesicles containing the Rab5 and Rab4 small GTPases (markers of the "short cycle"), whereas there was no colocalization with Rab11 (a marker of the "long cycle") or Rab7 (a marker of vesicles directed to endosomal/lysosomal compartments). Taken together, these data indicate that OTRs are capable of very efficient and complete resensitization due to receptor recycling via the short cycle. PMID- 19126787 TI - Effects of intrafetal IGF-I on growth of cardiac myocytes in late-gestation fetal sheep. AB - Intrafetal insulin-like growth factor (IGF)-I promotes cardiac hypertrophy in the late-gestation fetal sheep; whether these effects are sustained is unknown. IGF-I was infused for 4 days at 80 microg/h from 121 to 125 days of gestation, and its effects at 128 days, 3 days after the infusion stopped, were determined by comparison with untreated fetal sheep. After IGF-I treatment, fetal weights were similar to those in control fetuses but kidney weights were bigger (P < 0.05), as were spleen weights of male fetuses (P < 0.05). Cardiac myocytes were larger in female than male fetal sheep (P < 0.001). IGF-I increased male (P < 0.001) but not female myocyte volumes. IGF-I did not alter the proportions of uni- or binucleated right or left ventricular myocytes. Female fetal sheep had a greater proportion of binucleated cardiac myocytes than males (P < 0.05). IGF-I-treated fetuses had a slightly greater proportion of right ventricular nuclei in cell cycle phase G(2)/M and a reduced proportion of G(0)/G(1) phase nuclei (P < 0.1). Therefore, evidence for IGF-I-stimulated cardiac cell hyperplasia in fetal sheep in late gestation was limited. In conclusion, the greater sizes and larger proportion of binucleated cardiac myocytes in female fetal sheep suggest that myocyte maturation may occur earlier in females than in males. This may explain in part the male sex-specific responsiveness of cardiac hypertrophy to IGF-I in late gestation. If IGF-I-stimulated cardiomyocyte growth is accompanied by maturation of contractile function, IGF-I may be a potential therapeutic agent for maintaining cardiac output in preterm males. PMID- 19126788 TI - Evidence for substance abuse services and policy research: a systematic review of national databases. AB - We reviewed 39 national government- and nongovernment-sponsored data sets related to substance addiction policy. These data sets describe patients with substance use disorders (SUDs), treatment providers and the services they offer, and/or expenditures on treatment. Findings indicate the availability of reliable data on the prevalence of SUD and the characteristics of specialty treatment facilities, but meager data on financing and services. Gaps in information might be filled through agency collaboration to redesign, coordinate, and augment existing substance abuse and general health surveys. Despite noted gaps, these data sets represent an unusually rich set of resources for health services and policy research. PMID- 19126786 TI - Estrogen replacement enhances EDHF-mediated vasodilation of mesenteric and uterine resistance arteries: role of endothelial cell Ca2+. AB - Endothelium-derived hyperpolarizing factor (EDHF) plays an important role in the regulation of vascular microcirculatory tone. This study explores the role of estrogen in controlling EDHF-mediated vasodilation of uterine resistance arteries of the rat and also analyzes the contribution of endothelial cell (EC) Ca(2+) signaling to this process. A parallel study was also performed with mesenteric arteries to provide comparison with a nonreproductive vasculature. Mature female rats underwent ovariectomy, with one half receiving 17beta-estradiol replacement (OVX+E) and the other half serving as estrogen-deficient controls (OVX). Uterine or mesenteric resistance arteries were harvested, cannulated, and pressurized. Nitric oxide and prostacyclin production were inhibited with 200 microM N(G) nitro-l-arginine and 10 microM indomethacin, respectively. ACh effectively dilated the arteries preconstricted with phenylephrine but failed to induce dilation of vessels preconstricted with high-K(+) solution. ACh EC(50) values were decreased by estrogen replacement by five- and twofold in uterine and mesenteric arteries, respectively. As evidenced by fura-2-based measurements of EC cytoplasmic Ca(2+) concentration ([Ca(2+)](i)), estrogen replacement was associated with increased basal and ACh-stimulated EC [Ca(2+)](i) rise in uterine, but not mesenteric, vessels. These data demonstrate that EDHF contributes to endothelium-dependent vasodilation of uterine and mesenteric resistance arteries and that estrogen controls EDHF-related mechanism(s) more efficiently in reproductive vs. nonreproductive vessels. Enhanced endothelial Ca(2+) signaling may be an important underlying mechanism in estrogenic modulation of EDHF-mediated vasodilation in small resistance uterine arteries. PMID- 19126789 TI - Temporal profile of clinical signs and histopathologic changes in an F-344 rat model of kainic acid-induced mesial temporal lobe epilepsy. AB - Since there is limited information in the literature, the purpose of this study was to investigate clinical signs, morphology, and temporal progression of lesions from Days 3 to 168 in a kainic acid (KA)-induced Fischer-344 (F-344) rat model of mesial temporal lobe epilepsy (MTLE). Following a single KA subcutaneous dose of 9 mg/kg to young adult male rats, 95% survived, 93% exhibited status epilepticus, and 80% eventually developed spontaneous motor seizures. Histopathology included hematoxylin and eosin (H&E), autofluorescence, Fluoro Jade B, Timm's, ED-1/CD68, GFAP, doublecortin, and Ki-67. Neuronal degeneration occurred on Day 3 in the hippocampal CA1, CA3, and dentate hilar regions; amyg daloid and thalamic nuclei; and frontoparietotemporal, entorhinal and piriform cortices. Degeneration severity peaked on Day 6 and decreased progressively until Day 168. Aberrant mossy fiber (MF) sprouting was present in the inner molecular layer of dentate gyrus on Days 6-168. Microliosis and astrogliosis peaked on Day 28 and generally colocalized with the distribution of neuronal degeneration. Important correlates to human MTLE included induction of spontaneous seizures, more severe neuronal damage of CA1 than CA3 (in contrast to other animal models but similar to humans), hilar neuronal loss, activated microgliosis and astrogliosis, aberrant MF sprouting, and dentate granule cell neurogenesis. Aberrant MF sprouting prior to spontaneous motor seizures and reduced seizure frequency with a decrease in aberrant MF sprouting support the hypothesis that MF sprouts are necessary for spontaneous seizure generation and maintenance. PMID- 19126790 TI - Frequency of different estrous stages in purpose-bred beagles: a retrospective study. AB - The beagle is a monoestric, nonseasonal breeder with a long estrous cycle. Owing to lengthy stages in individual phases of the estrous cycle, limited group size, and typical group assignment focused on homogenized body weight, dogs in the same stage of the cycle can be inadvertently assigned to one treatment group in toxicity studies potentially leading to erroneous interpretation. This study was conducted to better understand the frequency of the different stages of the cycle and review the associated histological features. Histologic sections of reproductive tissues were reviewed from 102 control dogs from thirty-two GLP studies. The average age of dogs at necropsy was 14.38 months, and the mean terminal body weight was 6.87 kg. Based on histological classification, fifty five dogs were in anestrus, twenty-eight in diestrus, nine in estrus, five in proestrus, and five were classified as immature. Mean ovarian weights were higher in the estrus stage. This review indicates that more than 80% of the dogs in this study were in the anestrus-diestrus stage, and a small percentage of dogs were immature. Interpretation of drug-induced effects on the morphologic changes in the reproductive tract should be performed with due consideration given to the stage of the cycle and the potential for nonuniform assignment to drug treatment groups. PMID- 19126791 TI - Effects of p38 MAP kinase inhibitors on the differentiation and maturation of erythroid progenitors. AB - In rodents, p38 MAP kinase inhibitors (p38is) induce bone marrow hypocellularity and reduce reticulocyte and erythrocyte counts. To identify target cell populations affected, a differentiating primary liquid erythroid culture system using sca-1(+)cells from mouse bone marrow was developed and challenged with p38is SB-203580, SB-226882, and SB-267030. Drug-related alterations in genes involved at different stages of erythropoiesis, cell-surface antigen expression (CSAE), burst-forming unit erythroid (BFU-E) colony formation, and cellular morphology (CM), growth (CG), and viability were evaluated. CSAE, CM, and decreases in BFU-E formation indicated delayed maturation, while CG and viability were unaffected. Terminal differentiation was delayed until day 14 versus day 7 in controls. CSAE demonstrated higher percentages of sca-1(+)cells after day 2 and reduced percentages of ter119(+) cells after day 7 in all treated cultures. Real-time reverse transcriptase polymerase chain reaction revealed a transient delay in expression of genes involved at early, intermediate, and late stages of erythropoiesis, followed by rebound expression at later time points. Results demonstrate p38is do not irreversibly inhibit erythrogenesis but induce a potency dependent, transient delay in erythropoietic activity. The delay in activity is suggestive of effects on sca-1(+)bone marrow cells caused by alterations in expression of genes related to erythroid commitment and differentiation resulting in delayed maturation. PMID- 19126792 TI - Congenital intrahepatic vascular anomaly in a clinically normal laboratory beagle. AB - Routine necropsy examination of a clinically normal, ten-month-old male beagle dog identified disruption of the dorsal aspect of the right median liver lobe by a multiloculated, pale tan-white, thick-walled mass, with diffuse hypertrophy of the left and caudate liver lobes. The nature of the lesion was investigated using a variety of immunohistochemical and histochemical techniques. Histologically, the lesion was characterized by marked, focally extensive vascular hyperplasia and ectasia with lobular atrophy, periportal fibrosis, and biliary hyperplasia. The endothelium of proliferating vessels was positive for von Willebrands factor, and proliferating bile ducts were positive for pancytokeratin. Based on the gross and histologic appearance, this lesion was identified as a vascular hamartoma with arteriovenous fistula. It is important to note that this spontaneous lesion may occur in the laboratory beagle in the absence of clinical signs. PMID- 19126793 TI - Molecular characterization of trimellitic anhydride-induced respiratory allergy in Brown Norway rats. AB - To contribute to the hazard identification of low molecular weight (LMW) respiratory allergens, respiratory allergy induced by trimellitic anhydride (TMA) was characterized by whole genome analysis of lung tissue and blood proteomics in Brown Norway rats. Dermal sensitization (50% and 25% w/v) with TMA and an inhalation challenge of 15 mg/m(3) TMA-induced apneas, laryngeal inflammation, increased numbers of eosinophils, neutrophils and macrophages in bronchoalveolar lavage (BAL), and increased immunoglobulin E levels in serum and lung tissue. Whole genome analysis of lung, sampled 24 hours after challenge, showed expression changes of not only genes belonging to several Gene Ontology groups with up-regulation of inflammatory-associated genes and those associated with lung remodeling but also genes involved in downsizing these processes. Blood proteomics reflected activation of inflammation-inhibiting pathways. Unsensitized animals challenged with TMA exhibited also an increased number of macrophages in BAL, but gene expression in the above-mentioned gene pathways was unchanged or down-regulated. The authors conclude that parameters for lung remodeling can be a valuable tool in hazard identification of LMW respiratory allergens. PMID- 19126794 TI - Inorganic arsenic-induced intramitochondrial granules in mouse urothelium. AB - Based on epidemiological data, chronic exposure to high levels of inorganic arsenic in the drinking water is carcinogenic to the urinary bladder of humans. Recently, models have been developed involving transplacental administration of inorganic arsenic and subsequent administration of another substance that produces a low incidence of urogenital neoplasms. Administration of arsenite or arsenate in the diet or drinking water to five-to eight-week-old mice or rats rapidly induces urothelial cytotoxicity and regenerative hyperplasia. In mice administered arsenite, we observed eosinophilic intracytoplasmic granules present in the urothelial cells. These granules were not present in urothelial cells of untreated mice or in treated or untreated rats. By transmission electron microscopy, the granules were located within the mitochondrial matrix, that is, mitochondrial inclusions. Arsenic, primarily as arsenite, was present in partially purified mitochondria containing these granules. Cells containing the granules were not usually associated with degenerative changes. Lack of these granules in rats suggests that they are not necessary for inorganic arsenic induced urothelial cytotoxicity or hyperplasia. These granules have also been observed with exposures to other metals in other tissues and other species, suggesting that they represent a protective mechanism against metal-induced toxicity. PMID- 19126795 TI - Some aids to histological recognition of hyaline droplet nephropathy in ninety day toxicity studies. PMID- 19126796 TI - Issues in the design and interpretation of chronic toxicity and carcinogenicity studies in rodents: approaches to dose selection. Letter. PMID- 19126799 TI - Direct evidence for cortical suppression of somatosensory afferents during visuomotor adaptation. AB - Upon exposure to novel visuomotor relationships, the information carried by visual and proprioceptive signals becomes discrepant, often disrupting motor execution. It has been shown that degradation of the proprioceptive sense (arising either from disease or experimental manipulation) enhances performance when drawing with mirror-reversed vision. Given that the central nervous system can exert a dynamic control over the transmission of afferent signals, reducing proprioceptive inflow to cortical areas could be part of the normal adaptive mechanisms deployed in healthy humans upon exposure to novel visuomotor environments. Here we address this issue by probing the transmission of somatosensory afferents throughout the course of adaptation to a visuomotor conflict, by recording median nerve somatosensory evoked potentials. We show that early exposure to tracing with mirror-reversed vision is accompanied by substantial proprioceptive suppression occurring in the primary somatosensory cortex (S1). This proprioceptive gating is gradually alleviated as performance increases with adaptation, returning to baseline levels. Peripheral and spinal evoked potentials were not modulated throughout, suggesting that the gating acted to reduce cortico-cortico excitability directly within S1. These modulations provide neurophysiological evidence for flexibility in sensory integration during visuomotor adaptation, which may functionally serve to reduce the sensory conflict until the visuo-proprioceptive mapping is updated. PMID- 19126798 TI - Role of corticospinal suppression during motor preparation. AB - Behavior arises from a constant competition between potential actions. For example, movements performed unimanually require selecting one hand rather than the other. Corticospinal (CS) excitability of the nonselected hand is typically decreased prior to movement initiation, suggesting that response selection may involve mechanisms that inhibit nonselected candidate movements. To examine this hypothesis, participants performed a reaction time task, responding with the left, right, or both indexes. Transcranial magnetic stimulation was applied over the right primary motor cortex (M1) to induce motor-evoked potentials (MEPs) in a left hand muscle at various stages during response preparation. To vary the time of response selection, an imperative signal was preceded by a preparatory cue that was either informative or uninformative. Left MEPs decreased following the cue. Surprisingly, this decrease was greater when an informative cue indicated that the response might require the left hand than when it indicated a right hand response. In the uninformative condition, we did not observe additional attenuation of left MEP after an imperative indicating a right hand response. These results argue against the "deselection" hypothesis. Rather, CS suppression seems to arise from "impulse control" mechanisms that ensure that responses associated with potentially selected actions are not initiated prematurely. PMID- 19126800 TI - The morphology of supragranular pyramidal neurons in the human insular cortex: a quantitative Golgi study. AB - Although the primate insular cortex has been studied extensively, a comprehensive investigation of its neuronal morphology has yet to be completed. To that end, neurons from 20 human subjects (10 males and 10 females; N = 600) were selected from the secondary gyrus brevis, precentral gyrus, and postcentral gyrus of the left insula. The secondary gyrus brevis was generally more complex in terms of dendritic/spine extent than either the precentral or postcentral insular gyri, which is consistent with the posterior-anterior gradient of dendritic complexity observed in other cortical regions. The male insula had longer, spinier dendrites than the female insula, potentially reflecting sex differences in interoception. In comparing the current insular data with regional dendritic data quantified from other Brodmann's areas (BAs), insular total dendritic length (TDL) was less than the TDL of high integration cortices (BA6beta, 10, 11, 39), but greater than the TDL of low integration cortices (BA3-1-2, 4, 22, 44). Insular dendritic spine number was significantly greater than both low and high integration regions. Overall, the insula had spinier, but shorter neurons than did high integration cortices, and thus may represent a specialized type of heteromodal cortex, one that integrates crude multisensory information crucial to interoceptive processes. PMID- 19126802 TI - The cover. Feathered tunic with felines, birds, and fish. PMID- 19126801 TI - A 51-year-old woman with bipolar disorder who wants to quit smoking. AB - Smoking among patients with mental illness is a major and underappreciated public health problem. The case of Ms G, a 51-year-old woman with bipolar disorder who wishes to quit smoking, illustrates the importance and feasibility of smoking cessation in patients with psychiatric disorders. Persons with chronic mental illness and/or substance abuse constitute 22% of the US population yet are estimated to consume 44% of cigarettes. As many as 200,000 of the 435,000 annual deaths related to smoking in the United States are estimated to occur in this population. On average, patients with mental illness die 25 years earlier than the general population, and smoking is a major contributor to these premature deaths. In the past, mental health clinicians have tended not to address smoking cessation with their patients, but increasing evidence suggests that such reticence is unwarranted, as smoking cessation in this population is feasible. The approach to cessation should include standard interventions of counseling and pharmacotherapy, for which substantial evidence of efficacy exists in patients with and without mental illness. If patients with mental illness are to achieve wellness, smoking cessation must be an integral component of their treatment regimen. PMID- 19126803 TI - A piece of my mind. Drama in medicine? PMID- 19126804 TI - Studies examine inflammatory biomarker in prevention and prediction of heart disease. PMID- 19126805 TI - Safer therapies for newborns probed. PMID- 19126806 TI - Outcomes in the era of bare-metal stents vs the era of drug-eluting stents. PMID- 19126807 TI - Outcomes in the era of bare-metal stents vs the era of drug-eluting stents. PMID- 19126808 TI - Failure to report financial disclosure information. PMID- 19126809 TI - Primary androgen deprivation therapy in men with prostate cancer. PMID- 19126810 TI - Respirator tolerance in health care workers. PMID- 19126812 TI - How to use an article about genetic association: A: Background concepts. AB - This is the first in a series of 3 articles serving as an introduction to clinicians wishing to read and critically appraise genetic association studies. We summarize the key concepts in genetics that clinicians must understand to review these studies, including the structure of DNA, transcription and translation, patterns of inheritance, Hardy-Weinberg equilibrium, and linkage disequilibrium. We review the types of DNA variation, including single-nucleotide polymorphisms (SNPs), insertions, and deletions, and how these can affect protein function. We introduce the idea of genetic association for both single-candidate gene and genome-wide association studies, in which thousands of genetic variants are tested for association with disease. We use the APOE polymorphism and its association with dementia as a case study to demonstrate the concepts and introduce the terminology used in this field. The second and third articles will focus on issues of validity and applicability. PMID- 19126813 TI - Update: A 60-year-old woman with sexual difficulties. PMID- 19126814 TI - The Americans with Disabilities Act: shattered aspirations and new hope. PMID- 19126811 TI - Bilateral deep brain stimulation vs best medical therapy for patients with advanced Parkinson disease: a randomized controlled trial. AB - CONTEXT: Deep brain stimulation is an accepted treatment for advanced Parkinson disease (PD), although there are few randomized trials comparing treatments, and most studies exclude older patients. OBJECTIVE: To compare 6-month outcomes for patients with PD who received deep brain stimulation or best medical therapy. DESIGN, SETTING, AND PATIENTS: Randomized controlled trial of patients who received either deep brain stimulation or best medical therapy, stratified by study site and patient age (< 70 years vs > or = 70 years) at 7 Veterans Affairs and 6 university hospitals between May 2002 and October 2005. A total of 255 patients with PD (Hoehn and Yahr stage > or = 2 while not taking medications) were enrolled; 25% were aged 70 years or older. The final 6-month follow-up visit occurred in May 2006. INTERVENTION: Bilateral deep brain stimulation of the subthalamic nucleus (n = 60) or globus pallidus (n = 61). Patients receiving best medical therapy (n = 134) were actively managed by movement disorder neurologists. MAIN OUTCOME MEASURES: The primary outcome was time spent in the "on" state (good motor control with unimpeded motor function) without troubling dyskinesia, using motor diaries. Other outcomes included motor function, quality of life, neurocognitive function, and adverse events. RESULTS: Patients who received deep brain stimulation gained a mean of 4.6 h/d of on time without troubling dyskinesia compared with 0 h/d for patients who received best medical therapy (between group mean difference, 4.5 h/d [95% CI, 3.7-5.4 h/d]; P < .001). Motor function improved significantly (P < .001) with deep brain stimulation vs best medical therapy, such that 71% of deep brain stimulation patients and 32% of best medical therapy patients experienced clinically meaningful motor function improvements (> or = 5 points). Compared with the best medical therapy group, the deep brain stimulation group experienced significant improvements in the summary measure of quality of life and on 7 of 8 PD quality-of-life scores (P < .001). Neurocognitive testing revealed small decrements in some areas of information processing for patients receiving deep brain stimulation vs best medical therapy. At least 1 serious adverse event occurred in 49 deep brain stimulation patients and 15 best medical therapy patients (P < .001), including 39 adverse events related to the surgical procedure and 1 death secondary to cerebral hemorrhage. CONCLUSION: In this randomized controlled trial of patients with advanced PD, deep brain stimulation was more effective than best medical therapy in improving on time without troubling dyskinesias, motor function, and quality of life at 6 months, but was associated with an increased risk of serious adverse events. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00056563. PMID- 19126815 TI - The other Medical Home. PMID- 19126816 TI - Roentgenologic examination of the gallbladder (cholecystography): the article that launched a new era of radiology. PMID- 19126817 TI - Neurostimulation for Parkinson disease. PMID- 19126818 TI - JAMA patient page. Uterine fibroids. PMID- 19126819 TI - Evaluation of the Dental Health Component, of the Index of Orthodontic Treatment Need, by Swedish orthodontists. AB - The aim of the present study was to evaluate if orthodontists in Sweden agree with the grading in the Dental Health Component (DHC) of the Index of Orthodontic Treatment Need, to analyze if certain malocclusions are graded differently, and to determine if there are any background factors among Swedish orthodontists that could be related to their judgements. Two questionnaires were sent to 272 orthodontists in Sweden. In one questionnaire, they were asked to grade different malocclusions (defined in the DHC) according to the need for orthodontic treatment and in the other to provide details of their background. Eighty-one per cent (219) answered one or both questionnaires and 216 answered the questions concerning the need for orthodontic treatment. The Swedish orthodontists' judgements were then compared with the gradings in the DHC. Statistical analysis was undertaken using chi-square, likelihood ratio chi-square, phi coefficient, contingency coefficient, and Cramer's V tests. The result showed that almost all participating orthodontists agreed that grade 1 of the index indicated no need for treatment. For grade 2 (little need), the opinions differed, but still the majority were in agreement with the index. The judgements for malocclusions in grade 3 (borderline need) were widespread, but the majority considered 3a, 3c, and 3f to have a need or a great treatment need. The only statistical correlation with background factors was that female orthodontists graded 3f higher than borderline. For grades 4 and 5 (need and great need), the Swedish orthodontists thought that all the malocclusions required treatment except 4x, which the majority considered to be borderline. The participating Swedish orthodontists in this study graded fewer malocclusions in grade 3 and the majority were of the opinion that compared with the DHC, more malocclusions needed treatment. PMID- 19126820 TI - Dental Aesthetic Index scores and perception of personal dental appearance among Turkish university students. AB - The aim of this study was to investigate the relationship between Turkish university students' awareness of malocclusion, their satisfaction with their personal dental appearance, and the severity of their occlusal irregularities. The sample consisted of 841 randomly selected university students, 522 (62.1 per cent) males and 319 (37.9 per cent) females, aged 17-26 years (mean age, 21.91+/ 1.92 years). A pre-tested questionnaire was used to assess the subjects' awareness of malocclusion and satisfaction with their personal dental appearance; the actual severity of malocclusion was determined using the Dental Aesthetic Index (DAI). Statistical analysis was carried out using chi-square for gender differences and Spearman rank-order correlation coefficients for awareness of malocclusion, satisfaction with personal dental appearance, and DAI scores. Analysis of variance and univariate analysis, with age and gender as the independent variables, were further used to analyse the data. Weak but statistically significant, negative, correlations were found between awareness of malocclusion and satisfaction at the following DAI scores: or= 36 (r= -0.477, P<0.001), and a statistically insignificant, negative correlation at a score of 31-35 (r= -0.102, P>0.05). A statistically significant association was found between DAI and awareness of malocclusion (r= -0.305) and satisfaction with dental appearance (r=0.234). There were no significant associations between the department in which the individuals studied and the investigated variables (P>0.05). DAI scores were significantly higher for females. Generally, no statistically significant gender differences were found in relation to DAI scores, awareness, or satisfaction (P>0.05). The findings of this study showed that age had a significant effect on satisfaction and gender on DAI score variation. Females had a greater need for normative treatment except in the 20- to 22-year-olds, and satisfaction decreased with age. PMID- 19126821 TI - Journal impact factor: what it is and is not. PMID- 19126822 TI - Dosing nomograms: silos on a slope. AB - Developing dosing management guidelines or protocol approaches to pharmacotherapy can provide several benefits for standardization of care. Frequently, clinicians may need to incorporate multiple influencing factors to individualize management. In some cases, preestablished, standardized approaches may create barriers to individualized care, potentially categorizing patients into dosing categories (silos) that minimize their individualized needs. Approaches to managing direct thrombin inhibitor therapy have been recently explored and dosing approaches different from those provided in the prescribing information have been proposed. Considerations regarding individualized management of anticoagulant therapy, including the use of standardized dosing or monitoring protocols, are discussed here. PMID- 19126823 TI - Increases in C-reactive protein may predict recurrence of clozapine-induced fever. AB - OBJECTIVE: To report a case of recurrent clozapine-induced fever that was associated with a rise in C-reactive protein (CRP). CASE SUMMARY: A 73-year-old man with Lewy Body dementia was admitted for psychosis. He was treated with clozapine (initial dose 12.5 mg/day, titrated to 75 mg/day over 15 days). On day 15 of clozapine therapy, he developed a benign fever (maximum 38.4 degrees C) that was associated with a rise in the CRP level (3.96 mg/dL). The level normalized when clozapine was discontinued. However, when the patient was rechallenged with clozapine, the CRP level became elevated (4.36 mg/dL) after 3 days of therapy, with a subsequent recurrence of fever (38.7 degrees C). DISCUSSION: We postulate that the elevation in CRP levels and the subsequent fever were caused by the effects of clozapine on the cytokine system via interleukin-6 and tumor necrosis factor-alpha, resulting in an inflammatory response with an acute phase reaction. This case is unique, as it is the first reported in the literature associating a recurrence of clozapine-induced fever with the known immunomodulatory effects of clozapine on cytokines and CRP level. According to the Naranjo probability scale, this adverse effect is probably associated with clozapine. CONCLUSIONS: Clozapine-related fever is generally benign but difficult to assess and manage, as it can be confused with much more serious conditions. Further research is needed to study whether CRP is a useful tool in predicting and managing clozapine fever. PMID- 19126824 TI - Interaction between warfarin and black tea. PMID- 19126825 TI - Comparison of handwritten and electronically generated prescription drug instructions. PMID- 19126826 TI - Weight-based argatroban dosing nomogram for treatment of heparin-induced thrombocytopenia. AB - BACKGROUND: Manufacturer recommendations for argatroban use in the setting of heparin-induced thrombocytopenia (HIT) state that the dosage should be titrated to a goal activated partial thromboplastin time (aPTT) of 1.5-3 times the baseline aPTT. The lack of a clear dosing strategy with argatroban may result in delayed stabilization of aPTT. There are no published nomograms to guide the dosing of argatroban. OBJECTIVE: To study the anticoagulant effect and incidence of bleeding and thrombotic events in patients receiving argatroban, with doses determined using a weight-based nomogram. METHODS: Patients with suspected or documented HIT at an 800-bed teaching community hospital were prospectively treated, in a nonrandomized, nonblinded manner, with argatroban; dosage adjustments were made according to 1 of 2 variations of a dosing nomogram: standard or hepatic/critically ill. The primary outcomes were time to aPTT stabilization and percentage of patients whose aPTTs were within the therapeutic range of 45-90 seconds at 6, 12, 24, 48, 72, and 96 hours. Secondary outcomes were the percentage of patients whose aPTTs were subtherapeutic, supratherapeutic, or above the therapeutic threshold of 45 seconds at each time interval; incidence of thrombotic events; number of dosage adjustments to achieve stabilization; and number of major bleeding events. RESULTS: Fifty-one patients were prospectively treated using the standard (n = 34) and hepatic/critically ill (n = 17) nomograms. Mean time to aPTT stabilization was 16.25 hours with the standard nomogram and 27.05 hours with the hepatic/critically ill nomogram. The percentages of patients with aPTTs within the therapeutic range at each time interval were 82.4%, 82.4%, 88.2%, 96.4%, 100%, and 100% with the standard nomogram and 58.8%, 82.4%, 76.5%, 93.3%, 100%, and 90.9% with the hepatic/critically ill nomogram. There were no thrombotic events after the initiation of argatroban. Three cases of major bleeding occurred. CONCLUSIONS: The nomogram is an effective dosing tool for achieving and maintaining therapeutic levels of anticoagulation. PMID- 19126827 TI - Comment: frequency of medication errors with intravenous acetylcysteine for acetaminophen overdose. PMID- 19126828 TI - Contrasting measures of adherence with simple drug use, medication switching, and therapeutic duplication. AB - BACKGROUND: Multiple measures of adherence have been reported in the research literature and it is difficult to determine which is best, as each is nuanced. Occurrences of medication switching and polypharmacy or therapeutic duplication can substantially complicate adherence calculations when adherence to a therapeutic class is sought. OBJECTIVE: To contrast the Proportion of Days Covered (PDC) adherence metric with 2 variants of the Medication Possession Ratio (MPR, truncated MPR). METHODS: This study was a retrospective analysis of the North Carolina Medicaid administrative claims data from July 1999 to June 2000. Data for patients with schizophrenia (ICD-9-CM code 295.xx) who were not part of a health maintenance organization, not hospitalized, and not pregnant, taking at least one antipsychotic, were aggregated for each person into person-quarters. The numerator for PDC was defined as the number of days one or more antipsychotics was available and the MPR numerator was defined as the total days' supply of antipsychotics; both were divided by the total days in each person quarter. Adherence rates were estimated for subjects who used only one antipsychotic, switched medications, or had therapeutic duplication in the quarter. RESULTS: The final sample consisted of 25,200 person-quarters from 7069 individuals. For person-quarters with single antipsychotic use, adherence to antipsychotics as a class was: PDC 0.607, truncated MPR 0.640, and MPR 0.695 (p < 0.001). For person-quarters with switching, the average MPR was 0.690, truncated MPR was 0.624, and PDC was 0.562 (p < 0.001). In the presence of therapeutic duplication, the PDC was 0.669, truncated MPR was 0.774, and MPR was 1.238 (p < 0.001). CONCLUSIONS: The PDC provides a more conservative estimate of adherence than the MPR across all types of users; however, the differences between the 2 methods are more substantial for persons switching therapy and prescribed therapeutic duplication, where MPR may overstate true adherence. The PDC should be considered when a measure of adherence to a class of medications is sought, particularly in clinical situations in which multiple medications within a class are often used concurrently. PMID- 19126829 TI - The effectiveness of exercise in the management of post-natal depression: systematic review and meta-analysis. AB - BACKGROUND: Post-natal depression (PND) is a serious mental health problem that may be reduced by exercise. National Institute for Health and Clinical Excellence in England have recommended that health professions should consider exercise as a treatment for PND. OBJECTIVE: To evaluate the effectiveness of exercise in the management of PND. METHODS: Systematic review and meta-analysis of randomized controlled trials (RCTs). Data sources involved in the study are Cochrane Library (CENTRAL), MEDLINE, EMBASE, PsycINFO, Science Citation Index and Social Science Citation Index, CINAHL and SPORTDiscus. Review methods. Selection criteria are RCTs and quasi-RCTs that compared any type of exercise intervention with other treatments or no treatment in women with PND. Database searches and abstracts were reviewed independently by two authors. The Delphi criteria were used to assess the quality of included studies. Data were abstracted by two reviewers. Data synthesis is meta-analysis. Main outcome measure is post-natal depression. RESULTS: Five studies fulfilled our inclusion criteria. When compared with no exercise, exercise reduced symptoms of PND {SMD = -0.81 [95% confidence interval (CI): -1.53 to -0.10]}. The overall WMD in Edinburgh Post-natal Depression Scale score was -4.00 points (95% CI: -7.64 to -0.35). However, significant heterogeneity was found. The effect size was reduced considerably (non significant) when the trial that included exercise as a co-intervention with social support was excluded [SMD = -0.42 (95% CI: -0.90 to 0.05)] and heterogeneity was no longer present. CONCLUSIONS: Due to heterogeneity, it is uncertain whether exercise reduces symptoms of PND. Caution is also required when interpreting findings from the main analysis as only five small trials were included and CIs were wide. Further research is evidently required. PMID- 19126830 TI - Does a system of instalment dispensing for newly prescribed medicines save NHS costs? Results from a feasibility study. AB - BACKGROUND: In view of the increasing cost of general practice of drug prescribing, it is important to look at ways of reducing drug wastage and thereby improve the cost-effectiveness of prescribing. OBJECTIVE: To determine the costs and cost savings to the NHS of instalment dispensing for newly prescribed medicines and to quantify the extra costs incurred by patients. METHODS: Patients were randomized to receive either a normal (n = 103) or an instalment (n = 101) prescription. RESULTS: The difference between prescribed and dispensed drug costs in the intervention group was 0.98 UK pounds per patient (95% confidence interval 0.14-1.82 UK pounds), giving a 7% reduction in drug costs. The costs of the additional pharmacy time required to implement the intervention was calculated to be 5.02 UK pounds per patient. CONCLUSIONS: Introduction of a system of instalment dispensing produced savings in the general practice of drugs bill, but these were not large enough to offset additional costs for pharmacists. PMID- 19126831 TI - One of the central principles taught in research methods courses is that the wording of survey questions influences respondents' answers. PMID- 19126832 TI - The effects of survey question wording on rape estimates: evidence from a quasi experimental design. AB - The measurement of rape is among the leading methodological issues in the violence against women field. Methodological discussion continues to focus on decreasing measurement errors and improving the accuracy of rape estimates. The current study used a quasi-experimental design to examine the effect of survey question wording on estimates of completed and attempted rape and verbal threats of rape. Specifically, the study statistically compares self-reported rape estimates from two nationally representative studies of college women's sexual victimization experiences, the National College Women Sexual Victimization study and the National Violence Against College Women study. Results show significant differences between the two sets of rape estimates, with National Violence Against College Women study rape estimates ranging from 4.4% to 10.4% lower than the National College Women Sexual Victimization study rape estimates. Implications for future methodological research are discussed. PMID- 19126833 TI - If she is not a victim, does that mean she was not traumatized? Evaluation of predictors of PTSD symptomatology among college rape victims. AB - The issue of whether individuals can be traumatized by the experience of rape if they do not label the experience a victimization remains controversial. Indeed, there are conflicting findings with regard to the extent to which such unacknowledged victims experience posttraumatic symptoms. The goal of the current study was to evaluate acknowledgment status as a predictor of posttraumatic stress disorder symptomatology using structural equation modeling among a sample of 346 college rape victims. Results showed that whereas acknowledged victims reported more symptoms of posttraumatic stress disorder, acknowledgment status did not add to the prediction of posttraumatic stress disorder symptoms in a model including assault violence and other important predictors of these symptoms. PMID- 19126834 TI - Battered women who kill: the impact of expert testimony and empathy induction in the courtroom. AB - Mock jurors (N = 312) viewed a simulated trial involving a woman, charged with the murder of her abusive husband, entering a plea of not guilty by reason of self-defense. Expert testimony was varied using battered woman syndrome, social agency framework, or no expert testimony. Within expert testimony conditions, jurors were presented with opening and closing statements either including or not including instructions aimed at inducing empathy. Results indicate differences in gender and expert testimony for ratings of guilt as well as differences in gender, expert testimony, and empathy induction for perceptions of the defendant. PMID- 19126835 TI - A preliminary study of intimate partner violence among Nepali women in the United States. AB - Although there is a growing number of studies on intimate partner violence (IPV) in U.S. South Asian communities, the examination of IPV among Nepali women in the United States is still in the initial stage. The purpose of this study was to assess the prevalence of and vulnerabilities to IPV among 45 Nepali immigrant women residing in the New York metropolitan area. The findings demonstrated that 75.6% of women had been verbally insulted by their current partners, and 62.2% had to seek permission from their partners to go to their friends' or relatives' houses. PMID- 19126836 TI - Prevalence, types, and pattern of intimate partner violence among pregnant women in Lima, Peru. AB - This study assesses the prevalence, types, and pattern of intimate partner violence (IPV) during lifetime and current pregnancy for 2,392 women in Lima, Peru. The reported lifetime prevalence of any IPV (physical, sexual, or emotional) is 45.1%. For women who experienced abuse, the prevalence of lifetime physical, emotional, and sexual IPV is 34.2%, 28.4%, and 8.7%, respectively. Older (>/= 30 years), unmarried, employed, and economically disadvantaged women and those with little education are more likely to experience lifetime and pregnancy IPV. Efforts at universal antepartum IPV screening and appropriate interventions are needed to reduce the burden of violence experienced by pregnant women. PMID- 19126837 TI - Sympathetic skin responses evoked by different stimuli modalities in spinal cord injury patients. AB - OBJECTIVE: By using a combination of physiological and electrical peripheral nerve stimuli, the authors aimed to characterize the expected dysfunction of the circuits responsible for sympathetic skin response (SSR) in persons with spinal cord injury (SCI). METHODS: The authors examined SSR induced in the hand and foot in 50 SCI patients and 15 age-matched and gender-matched healthy volunteers. SSR was induced by deep inhalation, unexpected acoustic stimuli, brisk hand muscle contraction, and median and peroneal nerve electrical stimulation (PNS). RESULTS: SSRs to any stimulus modality were absent in hand and foot in patients with complete SCI above the T4 level. They were present in the hand and absent in the foot in complete SCI patients at levels between T4 and T11 for all stimuli modalities except PNS. The elicitability of SSR was lower with peroneal nerve stimulation than the other stimuli in hand and foot. The mean latency difference between SSRs of the hand and foot was significantly longer in patients than in controls, regardless of stimulus modality. The amplitude of SSR was larger in volunteers than in patients. CONCLUSION: SSR to various stimuli confirms the importance of supraspinal centers and the integrity of sympathetic descending pathways. Simultaneous recording of the SSR in the hands and feet provides information about the degree of sympathetic impairment possibly in the efferent pathway. To monitor spontaneous recovery or the efficacy of a drug or biological therapeutic intervention, changes in the latency delay between the hand and foot may be valuable. PMID- 19126838 TI - Influence of speed on walking economy poststroke. AB - BACKGROUND AND OBJECTIVE: Walking speed influences energy cost in healthy adults, but its influence when walking is impaired due to stroke is not clear. This study investigated the effect of manipulating walking speed on the energy economy of walking poststroke. METHODS: Sixteen persons with chronic stroke underwent a clinical examination, including several lower extremity impairment measures. consumption (VO(2)) was measured as they walked at their self-selected speed (Free), 20% slower (Slow), their fastest possible speed (Fastest), and 2 speeds between Free and Fastest speeds. VO(2) was normalized to body mass and speed, resulting in energy cost per meter walked (CW). RESULTS: A main effect for speed was observed (P = .00001), with faster than self-selected speeds showing greater relative economy as a whole. However, for 5 subjects with the fastest walking speeds (>1.2 m/s), there was a trend toward decreasing relative economy at speeds higher than self-selected speed (P = .18). There was a negative correlation between improvement in CW at the most economical speed and (a) Free speed (r = .857; P < .0001) and (b) lower extremity Fugl-Meyer scores (r = -.653; P = .006). CONCLUSIONS: For those poststroke whose fastest walking speed after stroke is below 1.2 m/s, walking economy improves when speed is increased above the self selected walking speed. The results suggest that for persons poststroke with very slow self-selected walking speeds, improvements in walking speed could be accompanied by improvements in walking economy if faster walking speeds can be attained through intervention. PMID- 19126839 TI - Age-related markers assayed at different developmental stages of the annual fish Nothobranchius rachovii. AB - Although short-lived vertebrates can serve as model animals for understanding the mechanism of aging, whether the annual fish Nothobranchius rachovii is suitable for studying aging remains an open question. In this study, histochemical, biochemical, and genetic techniques were used to determine the age-related markers at three different developmental stages of the annual fish N. rachovii. Histochemical studies revealed that the expression of senescence-associated beta galactosidase and accumulation of lipofuscin increased with age. In biochemical assays, lipid peroxidation and protein oxidation increased with age, whereas the activities of catalase, glutathione peroxidase, and superoxide dismutase decreased with age. Genetic analysis established that the activities of telomerase had no apparent relationship with age, but telomere lengths reduced with age from 11.5 +/- 1.98 to 3.58 +/- 0.74 kb. Taken together, these results indicate that the annual fish N. rachovii may be useful as an animal model for the study of aging. PMID- 19126841 TI - Airspace enlargement with airway cell apoptosis in klotho mice: a model of aging lung. AB - Homozygous mutant klotho (KL(-/-)) mice exhibit various characteristics resembling those of human aging, including emphysema. However, age-related changes of lungs have not been fully elucidated. Here, we investigated the structural, functional, biochemical, and cell kinetic alterations of lungs in KL( /-) mice at 2-12 weeks of age. Homogeneous airspace enlargement and decreased lung elastic recoil were observed in KL(-/-) mice with aging. The apoptotic cells in airway walls in KL(-/-) mice were approximately 6 times greater than those in wild-type (KL(+/+)) mice at 2 weeks of age. However, lipid peroxidation and elastase activity of lungs were not increased in KL(-/-) mice. Western blotting suggested that protein levels of epidermal growth factor (EGF) and phosphorylated extracellular signal-regulated kinase were decreased in KL(-/-) mice. These data suggest that significantly increased apoptosis of airway cells via inhibition of the EGF-dependent pathway may be involved in the development of the aging lungs in KL(-/-) mice. PMID- 19126840 TI - Principal component analysis reveals age-related and muscle-type-related differences in protein carbonyl profiles of muscle mitochondria. AB - Carbonyl-modified proteins are considered markers of oxidative damage caused by oxidative stress, aging, and disease. Here we use a previously developed capillary electrophoretic method for detecting femtomole (10(-15) mole) carbonyl levels in mitochondrial proteins that are size separated and profiled. For protein labeling, carbonyls were tagged with Alexa 488 hydrazine and amine groups in proteins with 3-(2-furoyl)quinoline-2-carboxaldehyde. Total mitochondrial protein carbonyl levels were statistically higher in fast- than in slow-twitch muscle of young Fischer 344 rats, and statistically higher in old than in young slow-twitch muscle. Even when some statistical comparisons of the total protein carbonyl levels would not reveal differences, principal component analysis (PCA) classified the carbonyl profiles into four distinct sample groups of different age and muscle types. In addition, PCA was used to predict that most age-related or muscle-type-related changes in carbonyl levels occur in proteins with a molecular weight between 9.8 and 11.7 kD. PMID- 19126842 TI - Diminished acute phase response and increased hepatic inflammation of aged rats in response to intraperitoneal injection of lipopolysaccharide. AB - Aging is associated with a deterioration of the acute phase response to inflammatory challenges. However, the nature of these defects remains poorly defined. We analyzed the hepatic inflammatory response after intraperitoneal administration of lipopolysaccharide (LPS) given to Fisher 344 rats aged 6, 15, and 22-23 months. Induction of the acute phase proteins (APPs), haptoglobin, alpha-1-acid glycoprotein, and T-kininogen was reduced and/or retarded with aging. Initial induction of interleukin-6 in aged rats was normal, but the later response was increased relative to younger counterparts. An exacerbated hepatic injury was observed in aged rats receiving LPS, as evidenced by the presence of multiple microabscesses in portal tracts, confluent necrosis, higher neutrophil accumulation, and elevated serum levels of alanine aminotransferase, relative to younger animals. Our results suggest that aged rats displayed a reduced expression of APPs and increased hepatic injury in response to the inflammatory insult. PMID- 19126843 TI - Cardiac and thermal homeostasis in the aging Brown Norway rat. AB - The cardiovascular and thermoregulatory systems are considered to be susceptible in the aged population, but little is known about baseline cardiac and thermoregulatory homeostasis in rodent models of aging. Radiotransmitters were implanted in male, Brown Norway rats obtained at 4, 12, and 24 months to monitor the electrocardiogram (ECG), interbeat interval (IBI), heart rate (HR), core temperature (Tc), and motor activity (MA). There was no significant effect of age on resting HR and MA. Daytime Tc of the 24-month-old rats was significantly elevated above those of the 4- and 12-month-old groups. Variability of the IBI was highest in the 24-month-old rats. The elevation in daytime Tc beginning around 8 months of age may be a physiological biomarker of aging and may be an important factor to consider in studies using caloric restriction-induced hypothermia to increase longevity. PMID- 19126844 TI - Guest editorial: linking thinking, walking, and falling. PMID- 19126845 TI - Age-associated effects of a concurrent cognitive task on gait speed and stability during narrow-base walking. AB - BACKGROUND: In older adults, changes in speed and stability during walking are associated with impaired balance and increased fall risk. Narrow-base walking requires increased frontal plane stability and can be used to assess postural control while walking. Performance of a concurrent cognitive task (dual task) may further increase the complexity of walking, potentially allowing identification of individuals with instability that is not detected under single-task conditions. The purpose of this study was to examine age-associated effects of a cognitive task on speed and frontal plane stability during narrow-base walking. METHODS: Thirty-four healthy adults participated, categorized by age: <65, 65-74, and > or =75 years. Participants walked at a comfortable pace within a narrow path under both single- and dual-task conditions. We examined spatiotemporal variables and frontal plane center of mass (CoM) parameters using a 13-segment biomechanical model. RESULTS: Increasing age (p <.001) and the performance of a concurrent cognitive task (p <.001) were both associated with decreased speed, with no interaction between these factors. Frontal plane CoM displacement and velocity increased with increasing age (both p <.001), but dual-task performance had no effect on these variables (both p >.450). CONCLUSIONS: Age-associated changes in both speed and stability are observed during narrow-base walking. Among this sample of healthy older adults, the addition of a concurrent cognitive task resulted in reduced speed, with no effect on frontal plane stability. Further research is needed to determine if dual-task, narrow-base walking is a sensitive and specific approach to identifying older adults at risk for falls. PMID- 19126846 TI - Dual-task decrements in gait: contributing factors among healthy older adults. AB - BACKGROUND: The factors that contribute to the dual tasking (DT) changes in performance that occur when older adults walk while simultaneously performing other tasks are not well known. We hypothesized that cognitive and motor reserve (e.g., executive function [EF], postural control, and walking abilities) and affect (e.g., anxiety, depressive symptoms) influence the DT decrements (DTDs) in gait. METHODS: Two hundred twenty-eight community-living, healthy older adults (mean: 76.2 +/- 4.2 years; 59% women) walked with and without DT, for example, subtracting 7s and phoneme monitoring. Mobility (e.g., the Dynamic Gait Index), cognitive function (e.g., memory, EF), and affect (e.g., Geriatric Depression Scale) were quantified. Bivariate and multivariate analyses identified factors associated with the DTD in gait speed (a general measure of locomotor function), swing time, (reflecting balance during gait), and swing time variability (a measure of stride-to-stride consistency). RESULTS: Gait speed and swing time decreased (p <.001) and swing time variability increased (became worse) (p <.001) during all DTs. The DTD in gait speed was correlated with comfortable walking gait speed, but not with tests of mobility or cognitive function. The DTD in swing time variability was correlated with EF, mobility, and affect (e.g., depressive symptoms). Much of the variance in the DTDs was unexplained. CONCLUSIONS: Usual walking abilities and cognitive function contribute to the DT effects on gait, but these relationships depend on specifics of the DT, the gait feature being studied, and the particulars of the cognitive domain. Meeting the everyday challenges of walking while dual tasking apparently relies on multiple factors including a consistent gait pattern and EF. PMID- 19126847 TI - Executive functions are associated with gait and balance in community-living elderly people. AB - BACKGROUND: Cognition influences gait and balance in elderly people. Executive functions seem to play a key role in this mechanism. Previous studies used only a single test to probe executive functions, and outcome measures were restricted to gait variables. We extend this prior work by examining the association between two different executive functions and measures of both gait and balance, with and without two different cognitive dual tasks. METHODS: This is a cross-sectional study with randomly selected community-living elderly people. Executive functions were tested with the Trail Making Test Parts A and B and the Stroop Color Word Test; memory with Cambridge Neuropsychological Test Automated Battery (CANTAB) subtests. Patients walked without and with two dual tasks (subtracting serial sevens and animal naming). Main outcomes focused on gait (velocity, stride length, and stride time variability), measured on an electronic walkway, and balance, measured as trunk movements during walking. Associations were assessed with multiple regression models. RESULTS: One hundred elderly people, with a mean age 80.6 years (range 75-93 years) participated. Both dual tasks decreased gait velocity and increased variability and trunk sway. Executive functions were associated with only stride length variability and mediolateral trunk sway during performance of animal naming as the dual task. Memory was not associated with the gait and balance variables. CONCLUSIONS: In community-living elderly people, executive functions are associated with gait and balance impairment during a challenging dual-task condition that also depends on executive integrity. Next steps will be to explore the value of executive functions in defining fall-risk profiles and in fall-prevention interventions for frail patients. PMID- 19126848 TI - Executive function and gait in older adults with cognitive impairment. AB - BACKGROUND: Cognitive impairment has been shown to predict falls risk in older adults. The ability to step accurately is necessary to safely traverse challenging terrain conditions such as uneven or slippery surfaces. However, it is unclear how well persons with cognitive impairment can step accurately to avoid such hazards and what specific aspects of cognition predict stepping ability in different patient populations. METHODS: Healthy older adults (NC), patients with Mild Cognitive Impairment with only memory impairment (MCI-EF) or memory and executive function impairments (MCI+EF) and early Alzheimer's patients (AD) were timed as they performed a stepping accuracy test with increasing cognitive demand (Walking Trail-Making Test; W-TMT), which required stepping on instrumented targets with either increasing sequential numbers (W-TMT A) or alternating sequential numbers and letters (W-TMT B). RESULTS: After accounting for age and baseline walking speed, the AD and MCI+EF groups were significantly slower than the NC and MCI-EF groups on the task with the highest cognitive demand, W-TMT B (interaction effect F = 6.781, p <.0001). No group differences were noted on the W-TMT A task that was less cognitively demanding. Neuropsychological measures of executive functioning were associated with slower W-TMT B performance, whereas memory, visual attention and visual spatial skills were not (adjusted R(2) = 0.42). CONCLUSIONS: Executive function is important for stepping performance, particularly under more complex environmental conditions. PMID- 19126849 TI - Egocentric and exocentric navigation skills in older adults. AB - BACKGROUND: Human spatial navigation can be conceptualized as egocentric or exocentric, depending on the navigator's perspective. Navigational impairment is known to occur in individuals with cognitive impairment, but less is known about navigational abilities in nondemented older adults. Our objective was to develop tests of navigation and study their cognitive correlates in nondemented older adults. METHODS: We developed a Local Route Recall Test (LRRT) to examine egocentric navigation and a Floor Maze Test (FMT) to assess exocentric navigation in 127 older adults without dementia or amnestic Mild Cognitive Impairment (MCI). Factor analysis was used to reduce neuropsychological test scores to three cognitive factors representing Executive Function/Attention, Verbal Ability, and Memory. We examined relationships between navigational tests and cognitive function (using both cognitive factors and the highest loading individual test on each factor) in a series of regression analyses adjusted for demographic variables (age, sex, and education), medical illnesses, and gait velocity. RESULTS: The tests were well tolerated, easy to administer, and reliable in this nondemented and non-MCI sample. Egocentric skills on the LRRT were associated with Executive Function/Attention (B = -0.650; 95% confidence interval [CI], 0.535 to -0.139) and Memory (B = -0.518; 95% CI, -4.893 to -0.063) factors. Exocentric navigation on the FMT was related to Executive Function/Attention (B = -8.542; 95% CI, -13.357 to -3.727). CONCLUSIONS: Our tests appear to assess egocentric and exocentric navigation skills in cognitively normal older adults, and these skills are associated with specific cognitive processes such as executive function and memory. PMID- 19126850 TI - Does inability to allocate attention contribute to balance constraints during gait in older adults? AB - BACKGROUND: Recent research has explored dual-task deficits during locomotion in older adults, yet the mechanisms underlying these deficits are poorly understood. In the current study, we examined one possible factor contributing to these deficits, the inability to flexibly allocate attention between two tasks. METHODS: Twelve healthy young adults and 12 healthy elderly adults performed obstacle avoidance while walking and an auditory Stroop task either alone or simultaneously. RESULTS: Using an attentional allocation index (AAI) to compare performance of healthy young and older adults and to measure the flexibility of allocation of attention, results showed a tendency in older adults toward a decreased ability to flexibly allocate their attention between the two tasks, with small AAI values. The decreased ability to allocate attention in older adults was found to be more prominent in the auditory Stroop task performance than in the obstacle avoidance task. CONCLUSION: This study suggests that an important factor contributing to decreased dual-task performance in older adults when simultaneously performing a postural and secondary cognitive task is a reduced ability to flexibly allocate attention between the two tasks, with the general ability to switch attention flexibly being predictive of the ability to adhere to a prioritized focus. PMID- 19126851 TI - Effect of competing attentional demands on perturbation-evoked stepping reactions and associated gaze behavior in young and older adults. AB - BACKGROUND: Rapid stepping reactions are a prevalent response to sudden loss of balance and are thought to play a crucial role in preventing falls. Previous dual task studies, involving concurrent performance of step reactions and a visuomotor tracking task, indicated that online visual attention was not required to guide the step, even when nearby objects increased demands for accurate foot movement. However, the planning and execution of the step apparently required attentional resources initially allotted to the tracking task. Reallocation of these resources ("attention switching") was delayed in older adults. The present study examined the influence of the competition for attentional resources by comparing trials performed with and without the concurrent task. METHODS: Unpredictable platform perturbations were used to evoke rapid forward stepping reactions in healthy young and older adults. Challenging obstacles and/or step targets increased demands for accurate foot motion in some trials. A concurrent tracking task was performed in half of the trials. RESULTS: Although participants looked down more frequently in the absence of the tracking task, the ability to clear the obstacle or land on the step target and other spatiotemporal features of the stepping reactions were largely unaffected. There was, however, one notable exception: In older adults, the duration and amplitude of the "anticipatory postural adjustment" that preceded foot lift were reduced in tracking trials, resulting in increased lateral center-of-mass motion. CONCLUSION: Impaired attention switching apparently compromised the control of lateral stability during stepping reactions in older adults, and may be an important contributor to increased risk of falling. PMID- 19126852 TI - Special article: gait measures indicate underlying focal gray matter atrophy in the brain of older adults. AB - OBJECTIVE: Our objective was to identify the spatial distribution of focal atrophy within mobility-related brain regions in relationship with quantitative gait characteristics. METHODS: Gray matter volume was obtained from 220 older adults (78.0 years old, 63% women, 77% white) for brain regions of five domains: motor (motor, sensorimotor and supplementary areas, basal ganglia, cerebellum), visuospatial attention (inferior and superior posterior parietal lobules), cognitive processing speed/executive control function (dorsolateral prefrontal cortex), memory (hippocampus), and motor imagery (parahippocampus, posterior cingulated cortex) domains. Spatial (step width, step length) and temporal (double support time) gait characteristics were measured using the GaitMat II instrumented walking surface. Multivariable linear regression models were adjusted for demographics, total brain volume, and peripheral (body mass index, ankle-arm index, arthritis, vibratory sensation) and central (markers of diffuse brain structural abnormalities and of brain function) risk factors for gait impairment. RESULTS: Shorter steps and longer double support times were associated with smaller sensorimotor regions and also with smaller frontoparietal regions within the motor, visuospatial, and cognitive processing speed domains. The associations between wider step and smaller pallidum and inferior parietal lobule were less robust. None of the gait measures were associated with the cerebellum or with regions of the memory or motor imagery domains. CONCLUSIONS: Spatial and temporal characteristics of gait are associated with distinct brain networks in older adults. Addressing focal neuronal losses in these networks may represent an important strategy to prevent mobility disability. PMID- 19126853 TI - Fatigue and function over 3 years among older adults. AB - BACKGROUND: Fatigue is a common complaint among older adults, but the association of fatigue with subsequent function is not well known. METHODS: This 3-year longitudinal study of older primary care patients evaluates the association of fatigue, operationalized as feeling tired most of the time, with functional status at baseline and over time. RESULTS: After adjustment for multiple potential confounders, participants who were tired at baseline had worse Short Form-36 Physical Performance Index scores, activity of daily living scores, and gait speeds. These functional deficits persisted throughout the follow-up period. CONCLUSIONS: Fatigue in older adults is associated with functional deficits that persist for years. Further research is needed to understand the causes of fatigue and to develop specific treatments for this serious symptom. PMID- 19126854 TI - Performance-based functional assessment in older hospitalized patients: feasibility and clinical correlates. AB - BACKGROUND: Functional evaluation is a cornerstone of multidimensional geriatric assessment; however, little is known of the clinical value of standardized performance-based assessment in the acute care setting. The aim of this study was to evaluate the clinical correlates and short-term predictive value of the Short Physical Performance Battery (SPPB) in older patients admitted to the hospital for an acute medical event. METHODS: We enrolled 92 women and men 65 years old or older who were able to walk, who had a Mini-Mental State Examination (MMSE) score > or =18, and who were admitted to the hospital with a clinical diagnosis of congestive heart failure, pneumonia, chronic obstructive pulmonary disease (COPD), or minor stroke. The SPPB was assessed at hospital admission and discharge. Self-report functional assessment included basic activities of daily living (ADL) and instrumental activities of daily living (IADL). Spearman's rank correlation coefficients and multivariable linear regression analyses were used to study the association of SPPB score and functional and clinical characteristics, including length of hospital stay. RESULTS: The mean age was 77.7 years (range 65-94 years), 49% were female, 64.1% had congestive heart failure, 16% COPD, 13.1% pneumonia, and 6.5% minor stroke. At hospital admission the mean SPPB score was 6.0 +/- 2.7. SPPB scores were inversely correlated with age, the severity of the index disease, and IADL and ADL difficulty 2 weeks before hospital admission (p <.01), and were directly correlated with MMSE score (p =.002). On average, SPPB score increased 1 point (+0.97, standard error of the mean = 0.2; p for paired t test <.001) from baseline to hospital discharge assessment. After adjustment for potential confounders, baseline SPPB score was significantly associated with the length of hospital stay (p <.007). CONCLUSION: In older acute care inpatients, SPPB is a valid indicator of functional and clinical status. SPPB score at hospital admission is an independent predictor of the length of hospital stay. PMID- 19126855 TI - Life course social and health conditions linked to frailty in Latin American older men and women. AB - BACKGROUND: Gender, social conditions, and health throughout the life course affect functional health in later life. This article addresses two specific hypotheses: i) life-course social and health conditions are associated with frailty; and ii) differential exposure and/or vulnerability of women and men to life-course conditions may account for gender differences in frailty. METHODS: Data originated from a cross-national survey of older adults living in five large Latin American cities. Frailty was defined as the presence of three or more of five criteria: unintentional weight loss (10 pounds during the past year), self reported exhaustion/poor endurance, weakness (grip strength), limitations in lower extremities, and low physical activity; a prefrail state was defined as the presence of one or two of the above criteria. Associations between frailty and social and health indicators were examined using a proportional odds ordinal logistic regression. RESULTS: Prevalence of frailty varied from 0.30 to 0.48 in women and from 0.21 to 0.35 in men. Childhood (hunger, poor health, and poor socioeconomic conditions), adulthood (little education and non-white-collar occupation), and current social conditions (insufficient income) were associated with higher odds of frailty in both men and women. Comorbidity and body mass index were related to frailty, but their effects differed in women and men. Male/female age-adjusted odds of frailty varied from 1.55 (Bridgetown) to 2.77 (Havana). Differential exposure and vulnerability partially explained differences between women and men. CONCLUSION: Theoretical models to explain gender and social differences in frailty should use a life-course perspective. PMID- 19126856 TI - Sleep in assisted living facility residents versus home-dwelling older adults. AB - BACKGROUND: Sleep problems among assisted living facility (ALF) residents are not well understood, and sleep-related differences between ALF residents and home dwelling older adults have not been examined. METHODS: We compared sleep patterns in 19 ALF residents to sleep patterns in 19 matched home-dwelling older people (age > or =65 years). All were participating in the follow-up portion of a longitudinal study of sleep and functional outcomes following post-acute rehabilitation. Sleep was assessed with the Pittsburgh Sleep Quality Index and 1 week of wrist actigraphy. RESULTS: By actigraphy, ALF residents awoke earlier in the morning and exhibited more nighttime awakenings compared to home-dwelling participants (06:50 hours +/- 1:29 hours vs 07:51 hours +/- 1:19 hours and 19.5 +/- 8.5 vs 12.9 +/- 11.4 awakenings, respectively). CONCLUSIONS: Larger studies are needed to confirm these initial findings that ALF residents have more disrupted sleep than do home-dwelling older persons, and to examine the functional and health consequences of poor sleep among ALF residents. PMID- 19126857 TI - Depression, antidepressants, and bone mineral density in a population-based cohort. AB - BACKGROUND: It is uncertain whether depression and antidepressant use are associated with decreased bone mineral density (BMD) and whether these relationships differ for men and women. METHODS: The study used a case-cohort design within the Baltimore Epidemiologic Catchment Area Study, a population based sample of adults that recently completed its 23-year follow-up. Depression was measured at four time points during the follow-up period by the Diagnostic Interview Schedule. Lower spine BMD was measured at the fourth wave by dual energy x-ray absorptiometry. The association of BMD with lifetime history of depression and antidepressant medication use was studied using linear regression with bootstrap standard errors. RESULTS: A history of depression was associated with lower spine BMD after controlling for age, sex, race, calcium intake, alcohol use, smoking status, level of physical activity, percent body fat, and antidepressant medication use (-0.140 g/cm(2); p <.002). After controlling for depression, antidepressant medication use was associated with decreased BMD in women but not in men (-0.218 g/cm(2); p <.016). CONCLUSIONS: A history of depression predicted decreased lumbar spine BMD in men and women, and antidepressant use predicted decreased BMD in women even after controlling for depression. The magnitude of the effect of depression on BMD was approximately equivalent to 1 standard deviation in BMD and was therefore clinically significant. Providers should be aware of the physiologic consequences of depression as well as the possible risks to bone strength associated with antidepressant use in older patients. PMID- 19126858 TI - The Baltimore Longitudinal Study of Aging (BLSA): a 50-year-long journey and plans for the future. PMID- 19126859 TI - 2008: Signaling breakthroughs of the year. AB - This year's signaling breakthroughs extended from protein crystals to cells and subcellular structures to whole genomes. Nominations included research on synapses in brains and in B cells; the evolution and regulation of signaling networks; the identification of a new class of plant hormones; insights into the causes and treatments of disease, such as cancer and schizophrenia; and a possible way to stay in shape while avoiding exercise. PMID- 19126860 TI - Drosophila toll pathway: the new model. AB - In Drosophila, recognition of microbe-specific molecules (such as bacterial peptidoglycans) activates serine protease cascades that converge to activate the Toll pathway. Recent data show that the serine protease Grass, which is activated downstream of pattern recognition receptors and was initially thought to be a component only of the Gram-positive bacteria-induced signaling cascade, is also required for the induction of the Toll pathway after fungal infection. Persephone, a serine protease known to be specifically activated by fungal proteases, was also found to be required for sensing Gram-positive bacterial proteases. Thus, Persephone serves as a sensor for microbial activities from both fungi and Gram-positive bacteria. With these new discoveries, a new model has been proposed for activation of the Drosophila Toll pathway. PMID- 19126862 TI - Computational implications of cooperative plasticity induction at nearby dendritic sites. AB - Recent studies have revealed that plasticity is not regulated independently at individual synapses but rather that there is cooperativity or associativity between nearby synapses in the dendritic tree of individual cortical pyramidal cells. Here, I summarize experimental results regarding such cooperative plasticity and its underlying mechanisms and consider their computational implications. PMID- 19126861 TI - MicroRNAs: opening a new vein in angiogenesis research. AB - Activation of the angiogenic program in endothelial cells is vital for normal embryonic development and for physiological angiogenesis in the adult. In addition, angiogenesis is an important therapeutic target: Formation of new blood vessels is desirable for regenerative purposes, such as during tissue healing or transplantation, but can be pathological, as in diabetic retinopathy and cancer. The response of the vascular endothelium to angiogenic stimuli is modulated by noncoding RNAs called microRNAs. The endothelial cell-specific microRNA microRNA 126 (miR-126) promotes angiogenesis in response to angiogenic growth factors, such as vascular endothelial growth factor or basic fibroblast growth factor, by repressing negative regulators of signal transduction pathways. Additional microRNAs have been implicated in the regulation of various aspects of angiogenesis. Thus, targeting the expression of microRNAs may be a novel therapeutic approach for diseases involving excess or insufficient vasculature. PMID- 19126863 TI - Zwilling-A and -B, two related myelin proteins of teleosts, which originate from a single bicistronic transcript. AB - Myelination, the ensheathment of axons by membranes of highly specialized glial cells, has been a crucial innovation during early vertebrate evolution. It enables high nerve signal conduction velocities, while maintaining nervous system size and energy requirements at moderate levels. Consequently, myelination has been conserved in all extant gnathostome vertebrates. In a genomewide mRNA expression screen, we identified several novel neural crest and myelin-specific transcripts in the zebrafish (Danio rerio). Here, we describe the characterization of two proteins, Zwilling-A and -B (ZwiA and ZwiB), which are exclusively expressed in myelinating glia of teleosts. They are structurally homologous and are translated from a common, bicistronic transcript. No similarities to sequences or domains of other proteins were detected. Analysis of phylogeny, genomic organization, and genomic syntenies suggests that the zwi gene has appeared soon after the teleost-specific genome duplication event and evolved under conservative selective pressure. We hypothesize that ZwiA and ZwiB serve important physiological functions in teleost myelin. PMID- 19126864 TI - The influence of demography and weak selection on the McDonald-Kreitman test: an empirical study in Drosophila. AB - The McDonald-Kreitman (MK) test, which compares the ratio of polymorphism to divergence at nonsynonymous and synonymous sites, is frequently used to detect adaptive evolution in protein-coding sequences. Because the two classes of sites share a common evolutionary history, the MK test is thought to be robust to most demographic factors. However, weak selection on nonsynonymous sites can bias the MK test, especially when a species' effective population size has not been constant. Here, we present an empirical analysis of the influence of demography on the MK test by comparing test results for a common set of 136 genes, including a set of sex-biased genes that shows a strong signal of adaptive evolution, in two Drosophila melanogaster populations: an ancestral population from Africa and a derived population from Europe. The latter has undergone a relatively recent bottleneck, which has reduced its effective population size. We find that the MK test has less power to detect positive selection in the European population for two reasons. First, the overall reduced level of standing variation decreases the statistical power of the test. Second, the segregation of slightly deleterious nonsynonymous mutations biases the MK test away from detecting positive selection. The latter effect is stronger for X-linked genes, which have experienced the greatest reduction in effective population size outside of Africa, and also leads to the underestimation of rates of adaptive protein evolution by multilocus implementations of the MK test. Interestingly, a subset of autosomal female-biased genes shows an increased signal of adaptive evolution in the European population. This is inconsistent with currently accepted demographic scenarios and may reflect female-specific changes in selective constraint following the colonization of non-African habitats. PMID- 19126865 TI - Balancing and directional selection at exon-2 of the MHC DQB1 locus among populations of odontocete cetaceans. AB - The diversity of exon-2 (peptide-binding region) of the DQB1 locus (Class II, major histocompatibility complex, MHC) was investigated on an extended sample of populations of three focal cetacean species (two sibling delphinid species and another in the same family). We tested the hypothesis that dolphin populations with a worldwide distribution across different habitats and geographic regions will be under differential selective pressure by comparing DQB1 variation with variation at neutral markers and by investigating putative functional residues within the exon-2 sequence at the population level. Variation at the DQB1 locus was not correlated to neutral differentiation (assessed by comparison with microsatellite DNA markers), and overall F(ST) values were significantly lower for the MHC locus, consistent with expectations for balancing selection. Measures of heterozygosity and d(n)/d(s) ratios were also consistent with balancing selection. However, outliers in the F(ST) comparisons and the analysis of putative functional residues suggested incidences of directional selection in local populations. PMID- 19126866 TI - Structural and evolutionary innovation of the heterodimerization interface between USP and the ecdysone receptor ECR in insects. AB - Understanding how the variability of protein structure arises during evolution and leads to new structure-function relationships ultimately promoting evolutionary novelties is a major goal of molecular evolution and is critical for interpreting genome sequences. We addressed this issue using the ecdysone receptor (ECR), a major developmental factor that controls development and reproduction of arthropods. The functional ECR is a heterodimer of two nuclear receptors: ECR, which binds ecdysteroids, and its obligatory partner ultraspirade (USP), which is orthologous to the retinoid X receptor of vertebrates. Both genes underwent a dramatic increase of evolutionary rate in Mecopterida, the major insect terminal group containing Dipteras and Lepidopteras. We therefore questioned the implication of this event in terms of coevolution of their dimerization interface. A structural comparison revealed a 30% larger ligand binding domain (LBD) heterodimerization surface in the Lepidoptera Heliothis when compared with basal insects, associated with a symmetrization of the interface, which is exceptional for nuclear receptors. Reconstruction of ancestral sequences and homology modeling of the ancestral Mecopterida ECR-USP reveal that this enlarged dimerization surface is a synapomorphy for Mecopterida. Furthermore, we show that the residues implicated in the new dimerization surface underwent specific evolutionary constraints in Mecopterida indicative of their new and conserved role in the dimerization interface. Most of all, the novel surface originates from a 15 degrees torsion of a subdomain of USP LBD toward its partner ECR, which is a long-range consequence of the peculiar position of a Mecopterida specific insertion in loop L1-3, located outside of the interaction surface, in a less crucial domain of the partner protein. These results indicate that the coevolution between ECR and USP occurred through a novel mechanism of intramolecular epistasis that will undoubtedly be generalized for other molecules because it uses flexibility of a less-constrained region of a protein to modify the structure of another, critical part of the molecule. PMID- 19126867 TI - The relationship between DNA replication and human genome organization. AB - Assessment of the impact of DNA replication on genome architecture in Eukaryotes has long been hampered by the scarcity of experimental data. Recent work, relying on computational predictions of origins of replication, suggested that replication might be a major determinant of gene organization in human (Huvet et al. 2007. Human gene organization driven by the coordination of replication and transcription. Genome Res. 17:1278-1285). Here, we address this question by analyzing the first large-scale data set of experimentally determined origins of replication in human: 283 origins identified in HeLa cells, in 1% of the genome covered by ENCODE regions (Cadoret et al. 2008. Genome-wide studies highlight indirect links between human replication origins and gene regulation. Proc Natl Acad Sci USA. 105:15837-15842). We show that origins of replication are not randomly distributed as they display significant overlap with promoter regions and CpG islands. The hypothesis of a selective pressure to avoid frontal collisions between replication and transcription polymerases is not supported by experimental data as we find no evidence for gene orientation bias in the proximity of origins of replication. The lack of a significant orientation bias remains manifest even when considering only genes expressed at a high rate, or in a wide number of tissues, and is not affected by the regional replication timing. Gene expression breadth does not appear to be correlated with the distance from the origins of replication. We conclude that the impact of DNA replication on human genome organization is considerably weaker than previously proposed. PMID- 19126868 TI - Selection for translational robustness in Buchnera aphidicola, endosymbiotic bacteria of aphids. AB - Its strong intergenerational bottlenecks and effectively asexual reproduction have led Buchnera aphidicola, the endocellular symbiotic bacterium of aphids, to spectacular evolutionary and genomic changes in comparison with its free-living bacterial cousins. These changes summarize into high fixation rates of mildly deleterious destabilizing mutations. This predicts a sharp decline of its fitness and the consequent early demise of this endosymbiotic bacterium. Its survival for hundreds of millions of years casts doubt on genetic drift as the sole evolutionary force and seeks further explanation. We identify in Buchnera selection to increase the robustness of proteins to misfolding translation errors. Translational robustness varies between Buchnera lineages and protein functional categories. Metabolic proteins have been under selection for translational efficiency, whereas evolutionary rates of proteins involved in fundamental cellular processes have been largely determined by selection for translational robustness. We detect the strongest signal of translational robustness in B. aphidicola Cinara cedri with a very similar pattern to that inferred for the most common symbiotic ancestor of Buchnera lineages. This indicates that B. aphidicola Cinara cedri lineage may have probably reached the minimum evolutionary stable gene composition for endosymbiotic lifestyle. The evolutionary patterns from the comparative genomic analyses of these endosymbionts support a paradoxically complex dynamic for apparently simple genomes. PMID- 19126869 TI - Dichotomous haplotypic lineages of the immunoproteasome subunit genes, PSMB8 and PSMB10, in the MHC class I region of a Teleost Medaka, Oryzias latipes. AB - Sequence comparison of the medaka, Oryzias latipes, major histocompatibility complex (MHC) class I region between two inbred strains, the HNI (derived from the Northern Population) and the Hd-rR (from the Southern Population), revealed a approximately 100 kb highly divergent segment encompassing two MHC class IA genes, Orla-UAA and Orla-UBA, and two immunoproteasome beta subunit genes, PSMB8 and PSMB10. To elucidate the genetic diversity of this region, we analyzed polymorphisms of the PSMB8 and PSMB10 genes using wild populations of medaka from three genetically different groups: the Northern Population, the Southern Population, and the China-West Korean Population. A total of 1,245 specimens from 10 localities were analyzed, and all the PSMB8 and PSMB10 alleles were classified into the N (fixed in the HNI strain) or the d (fixed in the Hd-rR strain) lineage. Polymerase chain reaction analysis of the region from PSMB8 to PSMB10 indicated that the two allelic lineages of these genes are segregating together constituting dichotomous haplotypic lineages. Both haplotypic lineages were identified in all three groups, although the frequency of d haplotypic lineage (73-100%) was much higher than that of N haplotypic lineage (0-27%) in all analyzed populations. The two allelic lineages of the PSMB8 gene showed curious substitutions at the 31st and 53rd residues of the mature peptide, which are likely involved in formation of the S1 pocket, suggesting that these alleles have a functional difference in cleavage specificity. These results indicate that the two medaka MHC haplotypic lineages encompassing the PSMB8 and PSMB10 genes are maintained in wild populations by a balancing selection. PMID- 19126870 TI - Possible diversifying selection in the imprinted gene, MEDEA, in Arabidopsis. AB - Coevolutionary conflict among imprinted genes that influence traits such as offspring growth may arise when maternal and paternal genomes have different evolutionary optima. This conflict is expected in outcrossing taxa with multiple paternity, but not self-fertilizing taxa. MEDEA (MEA) is an imprinted plant gene that influences seed growth. Disagreement exists regarding the type of selection acting on this gene. We present new data and analyses of sequence diversity of MEA in self-fertilizing and outcrossing Arabidopsis and its relatives, to help clarify the form of selection acting on this gene. Codon-based branch analysis among taxa (PAML) suggests that selection on the coding region is changing over time, and nonsynonymous substitution is elevated in at least one outcrossing branch. Codon-based analysis of diversity within outcrossing Arabidopsis lyrata ssp. petraea (OmegaMap) suggests that diversifying selection is acting on a portion of the gene, to cause elevated nonsynonymous polymorphism. Providing further support for balancing selection in A. lyrata, Hudson, Kreitman and Aguade analysis indicates that diversity/divergence at silent sites in the MEA promoter and genic region is elevated relative to reference genes, and there are deviations from the neutral frequency spectrum. This combination of positive selection as well as balancing and diversifying selection in outcrossing lineages is consistent with other genes influence by evolutionary conflict, such as disease resistance genes. Consistent with predictions that conflict would be eliminated in self-fertilizing taxa, we found no evidence of positive, balancing, or diversifying selection in A. thaliana promoter or genic region. PMID- 19126871 TI - Placenta-derived chymotrypsin-like protease (CLP) disturbs endothelial junctional structure in preeclampsia. AB - Placenta-derived chymotrypsin-like protease may contribute to endothelial activation in preeclampsia. In this study, we determined if placenta-derived chymotrypsin-like protease could disturb endothelial junctional integrity to promote endothelial permeability in preeclampsia. Confluent endothelial cells were cocultured with placental trophoblasts or treated with preeclampsia placenta conditioned medium. Endothelial junction protein vascular endothelial cadherin expression and distribution were examined by fluorescent staining of endothelial cells with or without depletion of chymotrypsin. The association of endothelial cell junction protein complex VE-cadherin/beta-catenin/p120 was examined by a combined immuno-precipitation and immuno-blotting assay. Our results showed that endothelial cells cocultured with preeclampsia trophoblasts or exposed to preeclampsia placental conditioned medium exhibited a discontinuous distribution and reduced expression of vascular endothelial cadherin at cell contact regions. Vascular endothelial cadherin and p120 were expressed in control endothelial cells, but reduced or lost in endothelial cells exposed to preeclampsia placental conditioned medium, suggesting that the junctional protein complex of VE cadherin/beta-catenin/p120 was disrupted in endothelial cells exposed to preeclampsia placental conditioned medium. We also observed that removal of trophoblasts from the coculture system and depletion of the protease from the preeclampsia placental conditioned medium could restore the dysregulated endothelial junction protein expression and distribution. Chymotrypsin also induced a dose dependent increase in endothelial monolayer permeability. We conclude that chymotrypsin-like protease released by the placenta is at least one important mediator responsible for disrupting endothelial cell integrity and inducing endothelial permeability in preeclampsia. PMID- 19126873 TI - Hematopoietic stem cell transplantation for core binding factor acute myeloid leukemia: t(8;21) and inv(16) represent different clinical outcomes. AB - We analyzed 338 adult patients with acute myeloid leukemia (AML) with t(8;21) and inv(16) undergoing stem cell transplantation (SCT) who were registered in the Japan Society for Hematopoietic Cell Transplantation database. At 3 years, overall survival (OS) of patients with t(8;21) and inv(16) was 50% and 72%, respectively (P= .002). Although no difference was observed when restricted to allogeneic SCT in first complete remission (CR; 84% and 74%), OS of patients with t(8;21) and inv(16) undergoing allogeneic SCT in second or third CR (45% and 86% at 3 years; P= .008) was different. OS was not different between patients in first CR who received allogeneic SCT and those who received autologous SCT for both t(8;21) AML (84% vs 77%; P= .49) and inv(16) AML (74% vs 59%; P= .86). Patients with inv(16) not in CR did better after allogeneic SCT than those with t(8;21) (70% and 18%; P= .03). Patients with t(8;21) and inv(16) should be managed differently as to the application of SCT. SCT in first CR is not necessarily recommended for inv(16). For t(8;21) patients in first CR, a prospective trial is needed to clarify the significance of autologous SCT and allogeneic SCT over chemotherapy. PMID- 19126872 TI - Leaky severe combined immunodeficiency and aberrant DNA rearrangements due to a hypomorphic RAG1 mutation. AB - The RAG1/2 endonuclease initiates programmed DNA rearrangements in progenitor lymphocytes by generating double-strand breaks at specific recombination signal sequences. This process, known as V(D)J recombination, assembles the vastly diverse antigen receptor genes from numerous V, D, and J coding segments. In vitro biochemical and cellular transfection studies suggest that RAG1/2 may also play postcleavage roles by forming complexes with the recombining ends to facilitate DNA end processing and ligation. In the current study, we examine the in vivo consequences of a mutant form of RAG1, RAG1-S723C, that is proficient for DNA cleavage, yet exhibits defects in postcleavage complex formation and end joining in vitro. We generated a knockin mouse model harboring the RAG1-S723C hypomorphic mutation and examined the immune system in this fully in vivo setting. RAG1-S723C homozygous mice exhibit impaired lymphocyte development and decreased V(D)J rearrangements. Distinct from RAG nullizygosity, the RAG1-S723C hypomorph results in aberrant DNA double-strand breaks within rearranging loci. RAG1-S723C also predisposes to thymic lymphomas associated with chromosomal translocations in a p53 mutant background, and heterozygosity for the mutant allele accelerates age-associated immune system dysfunction. Thus, our study provides in vivo evidence that implicates aberrant RAG1/2 activity in lymphoid tumor development and premature immunosenescence. PMID- 19126874 TI - Interventions to lower the glycemic response to carbohydrate foods with a low viscosity fiber (resistant maltodextrin): meta-analysis of randomized controlled trials. AB - BACKGROUND: The glycemic response to diet has been linked with noncommunicable diseases and is reduced by low-palatable, viscous, soluble fiber (1). Whether a palatable, low-viscous, soluble fiber such as resistant maltodextrin (RMD) has the same effect is unclear. OBJECTIVE: The objective was to assess evidence on the attenuation of the blood glucose response to foods by < or = 10 g RMD in healthy adults. DESIGN: We conducted a systematic review of randomized, placebo controlled trials with the use of fixed- and random-effects meta-analyses and meta-regression models. RESULTS: We found data from 37 relevant trials to April 2007. These trials investigated the attenuation of the glycemic response to rice, noodles, pastry, bread, and refined carbohydrates that included 30-173 g available carbohydrate. RMD was administered in drinks or liquid foods or solid foods. Placebo drinks and foods excluded RMD. Percentage attenuation was significant, dose-dependent, and independent of the amount of available carbohydrate coingested. Attenuation of the glycemic response to starchy foods by 6 g RMD in drinks approached approximately 20%, but when placed directly into foods was approximately 10% -- significant (P < 0.001) by both modes of administration. Study quality analyses, funnel plots, and trim-and-fill analyses uncovered no cause of significant systematic bias. Studies from authors affiliated with organizations for-profit were symmetrical without heterogeneity, whereas marginal asymmetry and significant heterogeneity arose among studies involving authors from nonprofit organizations because of some imprecise studies. CONCLUSIONS: A nonviscous palatable soluble polysaccharide can attenuate the glycemic response to carbohydrate foods. Evidence of an effect was stronger for RMD in drinks than in foods. PMID- 19126876 TI - Collaboration for the arts in healthcare: an opportunity. PMID- 19126880 TI - Nurses' experiences of practicing the HeartTouch technique for one month. AB - Stress plays a significant role in almost every aspect of health. Appraisal (thoughts and feelings) of a situation determines the level of stress experienced. PURPOSE: To describe the nurses' experiences of practicing the HeartTouch (HRTT) technique, an internal tool designed to help nurses change their thoughts and feelings, especially in stressful situations. METHOD: After an education session discussing the effects of thoughts and feelings on stress and health, nurses learn HRTT. After practicing HRTT for 1 month during their daily life, 48 nurses answer 7 open-ended questions about their experience. Content analysis is used to analyze the data. FINDINGS: Nurses notice physical, mental, and emotional benefits for themselves, patients, colleagues, and family members. Nurses feel less stressed and experience an increased sense of control and ability to notice and change their thoughts and feelings. Nurses also discuss meaningful connections with self, others, and a Higher Power and greater work satisfaction. PMID- 19126884 TI - Hold my hand . . . don't let go: moments of caring from a patient's perspective. AB - The author provides her perspective, as a patient, on the holistic nursing she received during the birth of her second child. The moments of caring and compassion she encountered are described. Four years later, the author reflects on the spiritual and emotional journey she has had as a result of this experience. PMID- 19126883 TI - Absorption: an individual difference to consider in mind-body interventions. AB - Given that mind-body interventions constitute a major portion of complementary and alternative medicine used by the public, it seems important to explore those human characteristics that may contribute to the efficacy of mind-body interventions in those who are most likely to benefit. One such characteristic, absorption, reflects an individual's cognitive capacity for involvement in sensory and imaginative experiences in ways that alter an individual's perception, memory, and mood with behavioral and biological consequences. Thus, one's level of absorption may potentially create differential treatment effects in mind-body intervention outcomes. Conducting practical clinical trials helps address the challenge of determining whether a specific mind-body modality intervention may be effective. Such trials may be strengthened by including measures of personality dimensions such as absorption. PMID- 19126886 TI - Substantiation and maltreatment rereporting: a propensity score analysis. AB - Although it is a widely used indicator, the use of substantiation in child welfare practice and research is not without critics. Much of this criticism concerns the ability of the substantiation disposition to distinguish between child protective services (CPS) investigations in which maltreatment occurs or does not occur. This study examined the relationship between substantiation and maltreatment rereporting using an analytic technique known as propensity score matching (PSM). Children with initially substantiated maltreatment reports were at significantly higher risk for rereporting than those with initially unsubstantiated reports, even after matching the two groups on propensity scores based on several demographic and case characteristics. Although additional study using PSM on other samples is warranted, this evidence supports the predictive validity of the substantiation disposition and its continued use as one factor to consider when allocating limited post-investigation services. PMID- 19126887 TI - Allegory of the cave: on the theme of substantiation. PMID- 19126888 TI - A commentary on mandatory reporting legislation in the United States, Canada, and australia: a cross-jurisdictional review of key features, differences, and issues. AB - Mathews and Kenny recently reviewed the current state of mandatory reporting laws in Australia, Canada, and the United States. The purpose of this article is to draw attention to existing differences between these countries regarding exposure to domestic violence (EDV) and to discuss EDV as a specific and detrimental form of child abuse. PMID- 19126889 TI - Intra-operative somatosensory-evoked potential monitoring. AB - PURPOSE: To evaluate the usefulness of intra-operative somatosensory-evoked potentials (SSEPs) in monitoring spinal cord status via the posterior tibial nerve. METHODS: 84 men and 28 women aged 16 to 66 years (72% were aged 20 to 40 years) with spinal trauma (63 in the lumbar and 49 in the thoracic spine) underwent posterior instrumentation and fusion using bone grafts. All 63 patients with lumbar spinal injury and 35 of the patients with thoracic spinal injury were treated with pedicular screws. The remaining 14 patients had their thoracic spinal injury fixed with sublaminar wires. Cortical scalp recordings were used. Baseline tracings were obtained prior to surgical intervention and after establishment of anaesthesia. If changes persisted for more than 15 to 20 minutes or if they did not show definite signs of resolution, event reversal was considered. RESULTS: Of the 112 patients, 74 (66%) had no changes in Cz-Fpz patterns and neurological status, whereas 14 (13%) showed improved patterns (2 of them had the same neurological status postoperatively) and 24 (21%) displayed deteriorated patterns prompting intervention. Of the 24 patients prompting intervention, 20 improved substantially (19 had no new neurological deficits and one had deteriorated neurological status) and 4 improved minimally (2 had no new deficit and 2 had new deficits), with 88% sensitivity and 78% specificity. 15 patients were true-positives with an identifiable cause; 21 were false-positives with no neurological deterioration or recognisable cause. CONCLUSION: Intra operative SSEP monitoring helps identify acute neurological and systemic (hypoxia or hypotension) impairment and enables prompt correction. This makes surgery available to high-risk patients and enables surgeons to carry out more extensive procedures. It also provides valuable documentation in the event of medico-legal dispute. PMID- 19126890 TI - Anterior versus modified combined instrumentation for burst fractures of the thoracolumbar spine: a biomechanical study in calves. AB - PURPOSE: To compare stability after anterior instrumentation alone versus modified combined anterior and posterior instrumentation for burst fractures of the thoracolumbar spine in calves. METHODS: Thoracolumbar spines of 10 calves were used. An axial compression force was applied on each specimen using a material-testing machine, until there was a burst fracture at T12 or L1. Five specimens were fixed with anterior instrumentation alone, using 2 rods connected by 2 screws above and 2 screws below the fractured vertebra plus one tranverse connector. Another 5 were fixed with our modified technique of combined anterior and posterior instrumentation. This entailed one rod connected with one screw above and one screw below the fractured vertebra anteriorly, and another rod connected with one transpedicular screw above and one transpedicular screw below the fractured vertebra posteriorly. After instrumentation, the experiment was conducted again on each specimen and the compressive stiffness and vertebral height loss between the 2 groups compared. RESULTS: The mean compressive stiffness was significantly greater after modified combined anterior and posterior instrumentation than anterior instrumentation alone (5508 vs 2888 N, p=0.0256), whereas the respective vertebral height losses were 37 and 33 mm (p=0.3808). CONCLUSION: Our modified technique of combined anterior and posterior instrumentation provides greater stability than traditional anterior instrumentation alone. PMID- 19126891 TI - Modified rush pin technique for two- or three-part proximal humeral fractures. AB - PURPOSE: To report the outcomes of modified Rush pin fixation for proximal humeral fractures. METHODS: 42 men and 20 women aged 19 to 94 (mean, 64) years with 2- or 3-part proximal humeral fractures underwent reduction and fixation using the modified Rush pin technique. 11 patients died from reasons unrelated to the surgery. RESULTS: Of 40 (out of 51) patients completing a subjective functional assessment using an Oxford Shoulder Questionnaire, 28 (70%) had 2-part and 10 (25%) had 3-part displaced fractures, and the remaining 2 (5%) had fracture-dislocations (one being 2-part and one 3-part). 25 (63%) patients were very satisfied (including one with a 3-part fracture after 6 months of rehabilitation), 7 (17%) were moderately satisfied, and 8 (20%) were not satisfied. There were 8 complications, including pin cut-out from the proximal fragment (n=2), proximal pin migration (n=2), distal pin migration (n=1), cortical perforation during surgery (n=1), mild ulnar nerve symptoms (n=1). No patients had non-union, myositis ossificans, avascular necrosis of the humeral head, or axillary nerve injury. CONCLUSION: The modified Rush pin fixation minimises tissue dissection; the implants are cheap and readily available; and the technical expertise is easily learnt. This technique is a suitable alternative of fixing proximal humeral fractures, especially in the elderly. PMID- 19126892 TI - Acute invaginating docking for infected non-unions of the humerus. AB - PURPOSE: To assess the efficacy of acute invaginating docking for infected non unions of the humerus. METHODS: Eight men and 3 women aged 17 to 59 years with infected non-unions of the humerus underwent acute invaginating docking with shortening and Ilizarov fixation. RESULTS: The mean shortening was 2.9 cm owing to additional debridement. The mean time to external fixator removal was 14.9 (range, 8-28) weeks. The final bone result was excellent in 1, good in 8, and fair in 2. The functional result was excellent in 7 and good in 4. CONCLUSION: This modality enables simultaneous treatment of infected non-unions, axial alignment, vascularity, stability, and function. PMID- 19126893 TI - Dynamic movement of the long head of the biceps tendon in frozen shoulders. AB - PURPOSE: To assess the importance of the dynamic movement of the long head of the biceps (LHB) tendon for treatment of frozen shoulder. METHODS: 87 consecutive patients (with 88 frozen shoulders) aged 36 to 77 (mean, 54) years underwent arthroscopic capsular release by a single surgeon. Preoperative treatments included rehabilitation, steroid and/or hyaluronic acid injections. The inclusion criteria were severe night pain, no improvement of flexion and external rotation, and poor response to rehabilitation for at least 6 months. Shoulders were divided into 3 types; types A/B/C indicate slight/moderate/severe degree of synovitis and adhesion of the LHB tendon to the rotator interval. 23 shoulders were type A, 26 type B, and 39 type C. 18 of the 39 type-C shoulders were controls with release of the capsule alone but not the LHB tendon. Patients were followed up for a mean of 21 (range, 12-35) months. Changes in the American Shoulder and Elbow Surgeons (ASES) score, range of movement, and muscle strength (flexion and external rotation) among types A, B, C, and controls were compared. RESULTS: The severity of the adhesion of the LHB tendon to the rotator interval was associated with the ASES score. In all adhesion types, muscle strength and the range of movement in flexion, external rotation, and internal rotation improved postoperatively. CONCLUSIONS: Arthroscopic capsular release for adhesion of the LHB tendon to the rotator interval improves the sliding movement and thereby shoulder function. PMID- 19126894 TI - Radiological features of osteoarthritis of the acromiclavicular joint and its association with clinical symptoms. AB - PURPOSE: To determine whether increasing age is associated with increased radiological features of osteoarthritis of the acromioclavicular joint (ACJ) in a general population, and whether clinical symptoms correlate with radiological features. METHODS: Anteroposterior and axillary shoulder radiographs of 240 patients aged 20 to 80 years were randomly selected. The presence of stigmata of osteoarthritis of the ACJ including sclerosis, cysts, lysis, and osteophytes were recorded, and the width of the ACJ was measured. To determine the correlation between clinical symptoms and radiological features, the same radiological features were assessed for 100 further patients who had undergone either arthroscopic subacromial decompression (ASD) alone (n=50) or ASD plus ACJ excision (n=50, age-matched controls) based on clinical examination. RESULTS: Radiological features of osteoarthritis of the ACJ increased significantly with increasing age but were not related to gender or the side affected. Of the 10 features, only medial acromial sclerosis and superior clavicular osteophytes were more prevalent in patients with ASD plus ACJ excision than in those with ASD alone (p=0.016). The sensitivity, specificity, positive and negative predictive values of these features were poor. Therefore, clinical symptoms were not associated with radiological features of osteoarthritis of the ACJ. CONCLUSION: Radiological features should only be used as an adjunct in the decision to excise the ACJ. A thorough clinical examination is crucial in the assessment of ACJ pathology. PMID- 19126895 TI - Staple versus locking compression plate fixation after lateral closing wedge high tibial osteotomy. AB - PURPOSE: To compare the results of staple versus locking compression plate fixation after closing wedge high tibial osteotomy. METHODS: A group of 23 patients (24 knees) who underwent box high tibial osteotomy and staple fixation was compared with another group of 19 patients (22 knees) who underwent a similar procedure but with locking compression plate fixation. Both groups were followed up for 3 years. The range of movement, Hospital for Special Surgery (HSS) Knee Score, time to full weight bearing, incidence of delayed union, femorotibial angle, and stage of osteoarthritis were compared. RESULTS: At 6 months after the operation, the median HSS score and the proportion of patients with excellent or good scores were significantly higher in the locking compression plate than the staple fixation group (76 vs 62, p=0.003; 75% vs 42%, p=0.0354), but not at one and 3 years. The range of movement was significantly greater in the locking compression plate fixation group in the short term (6 weeks, 3 and 6 months), but not after one year. The median time to full weight bearing was significantly shorter in the locking compression plate fixation group (86 vs 116 days, p<0.001). There were fewer delayed unions in the locking compression plate fixation group but not significantly (1 vs 5, p=0.198), possibly because of the small numbers involved. There was no difference, within the limits of measurement error, in the femorotibial angle or correction loss between the 2 groups. CONCLUSION: Locking compression plate fixation obviates the use of plaster casts, enables early mobilisation and bone union, and reduces the numbers with delayed union and the time to full weight bearing. Longer-term studies are needed to evaluate its effect on revarisation and arthropathy. PMID- 19126896 TI - Avulsion fractures of the tibial tubercle. AB - PURPOSE: To retrospectively study the epidemiology and treatment of acute avulsion fractures of the tibial tubercle in 12 patients. METHODS: Records of 12 patients aged 11 to 17 (mean, 14) years with avulsion fractures of the tibial tubercle presenting to our hospital from April 1998 to September 2004 were studied. Patient age, gender, involved side, injury mechanism, clinical and radiographic records, treatment, complications, and outcomes were reviewed. RESULTS: Eight patients injured the right side, 3 the left side, and one both sides. They all engaged in sports or active play during the injury. Two fractures were type IA, one type IB, 2 type IIB, 5 type IIIA, 2 type IIIB, and one type IV. Three patients with type-I fractures and one with a type-IIB fracture were treated with closed reduction and cast immobilisation for 3 to 4 (mean, 3.8) weeks. The remaining 9 fractures were treated with open reduction and internal fixation. The mean follow-up period was 39 (range, 23-59) months. No complications were noted. Ten patients had excellent results and 2 had fair results. CONCLUSION: Closed reduction and cast immobilisation for minimally displaced fractures, and open reduction and internal fixation for displaced fractures resulted in favourable outcomes. PMID- 19126897 TI - Morphologic differences of the distal femur between Caucasian and Japanese women. AB - PURPOSE: To compare the morphology of the distal femur between Caucasian and Japanese women. METHODS: 30 Caucasian women aged 41 to 84 (mean, 67) years and 70 Japanese women aged 54 to 86 (mean, 70) years who underwent total knee arthroplasty for osteoarthritis were randomly selected. Morphologic measurements of the distal femur were compared using lateral radiographs. Both race and height influenced the morphology. To adjust for the influence of height on morphology, each measurement was divided by the patient's height and the ratios were compared. RESULTS: Caucasian women were generally taller and heavier (p<0.001) and had higher body mass index (p=0.03) than the Japanese women. Each morphologic measurement of the distal femur was significantly longer in the Caucasian women. In both groups, anteroposterior width of the condyle correlated more with height than weight. In women of equal height, the anteroposterior and metaphyseal widths of the femur and the anterior and resected condyles were longer in Caucasian women, but the posterior condyle was longer in Japanese women. CONCLUSION: Both the size of the femur and the anterior and posterior condyles are significantly larger in Caucasian than Japanese women. PMID- 19126898 TI - Inter- and intra-observer errors in identifying the transepicondylar axis and Whiteside's line. AB - PURPOSE: To assess inter- and intra-observer errors in identifying the transepicondylar axis and Whiteside's line in a cadaveric model mimicking total knee arthroplasty. METHODS: Four cadaveric knees with intact soft tissues were used. The knees were exposed anteriorly using the Insall approach, with the patella everted laterally. Three observers (2 surgeons and one trainee) took turns to identify the anatomic landmarks of the transepicondylar axis and Whiteside's line. Each observer repeated the process 20 times. Each identification was photographed and referenced with the true values obtained from the knees after they were stripped of all soft tissue. Inter- and intra-observer errors in the anatomic landmarks were compared. RESULTS: Inter-observer error was significant with both the transepicondylar axis and Whiteside's line (p<0.001, one-way ANOVA). The intra-observer variation was greater for Whiteside's line than the transepicondylar axis (standard deviation, 4.2 vs 2.5 degrees). The maximum potential errors in the transepicondylar axis and Whiteside's line were 13 degrees and 24 degrees, respectively. CONCLUSION: The accuracy of rotational alignment of the transepicondylar axis and Whiteside's line were operator dependent, and their intra-operative reproducibility was low. PMID- 19126899 TI - Anatomic study for pubic medullary screw insertion. AB - PURPOSE: To study the anatomy of the pubic ramus and adjacent structures in 160 Japanese to establish a safer pubic screw fixation technique. METHODS: 80 male and 80 female Japanese aged 16 to 89 (mean, 50) years (10 persons in each decade of age) underwent 3-dimensional computed tomographic scanning of their pelvises. The angle at which the screw should be targeted, the appropriate length of the screw, the size of the canal for screw insertion, and the proximity to the bladder, iliac artery, and iliac vein were determined. Correlations between the canal diameters (of the acetabular, base, and parasymphyseal areas) and body features (age, height, and weight) were analysed. RESULTS: In men and women respectively, the appropriate mean screw length was 124.6 and 123.8 mm; the guide wire should be targeted at a mean of 66 degrees and 67 degrees cephalad and 54.1 degrees and 55.9 degrees laterally for insertion of a retrograde pubic screw; the minimum distances from the pubis to the bladder/iliac artery/ iliac vein were 0 and 0 mm/4.9 and 4.6 mm/0.8 and 0.2 mm. In both men and women, the canal diameters at the base were positively correlated to weight. In women, the canal diameters at the parasymphyseal area were correlated to height and weight. Canal diameters at the acetabulum were not correlated to height and weight. CONCLUSION: Pubic screw fixation may be potentially disastrous (owing to joint penetration and iliac vein injury) and should be performed with caution. When the canal diameter at the acetabulum is extremely narrow, plate fixation, computer-assisted surgery, or changing to a smaller-diameter screw is recommended. PMID- 19126900 TI - Prosthesis design and stress profile after hip resurfacing: a finite element analysis. AB - PURPOSE: To evaluate the effect of prosthesis design on stress profile in the proximal femur after hip resurfacing. METHODS: The von Mises stress profile of the native femur was simulated and compared with that of resurfaced femurs using various prosthetic materials (titanium, cobalt-chrome, ceramic), stem lengths (normal, half, short, and no stem), and femoral head coverage (shell size) [260 degrees, 220 degrees, 180 degrees, and 140 degrees]. RESULTS: Hip resurfacing altered the stress profile of the cancellous (but not cortical) bone of the femoral neck. Maximal cortical stresses were observed at the posterior half of the medial femoral neck. The stress profile of the native femur was most similar to that of the resurfaced femur made of titanium, with a short or no stem and 260 degrees of femoral head coverage (shell size). CONCLUSION: Optimising prosthesis design by minimising biomechanical alterations seems a valid approach to achieving favourable long-term outcomes. Cadaveric and in vivo studies are needed to confirm the clinical relevance and feasibility. PMID- 19126901 TI - Pelvic Ewing's sarcoma: a review from Scottish Bone Tumour Registry. AB - PURPOSE: To review various treatment modalities on outcome of patients with pelvic Ewing's sarcoma. METHODS: Between January 1948 and December 2004, 18 male and 15 female patients aged 3 to 48 (mean, 20) years with pelvic Ewing's sarcoma were retrospectively reviewed from the Scottish Bone Tumour Registry. RESULTS: The commonest site involved was the ilium. The main presenting symptom was pain, followed by swelling and restricted hip movements. Treatments included surgery, radiotherapy, chemotherapy, or any of them in combination. Patients who had no metastasis at presentation (p<0.01) and underwent chemotherapy with ifosfamide (p<0.01) had significantly longer survival. Age (p=0.09), gender (p=0.95), delay in presentation (p=0.31), tumour site (p=0.9), surgery (p=0.73), and radiotherapy (p=0.23) were not predictive of survival in the univariate analysis. The 5-year event-free survival rate in patients with no metastasis at presentation was 35%. CONCLUSION: Intense multi-agent neo-adjuvant chemotherapy including ifosfamide followed by wide excision and postoperative radiation are recommended. Local therapy should not take precedence over or interfere with systemic chemotherapy. PMID- 19126902 TI - Microbiology of infected arthroplasty: implications for empiric peri-operative antibiotics. AB - PURPOSE: To classify the spectrum and antibiotic susceptibility of bacteria isolated from infected hip and knee arthroplasty specimens, and to recommend appropriate empiric peri-operative antibiotics. METHODS: From January 1999 to August 2006, specimens from revision hip and knee arthroplasties (with or without suspected infection) were routinely collected for identifying possible organisms and their susceptibility patterns. During the period, 147 patients had positive specimens yielding 248 micro-organisms (from 195 tissue specimens, 43 fluid specimens, and 10 swabs). 140 isolates were from hips and 108 from knees. RESULTS: Most isolates were Gram-positive; their distribution was similar in hip and knee specimens. Of the 248 micro-organisms isolated, Staphylococcus was the most common genus encountered (131, 53%), followed by Gram-negative isolates (24%). 88% of Gram-negative organisms were detected within 48 hours of inoculation and 94% of Gram-positive organisms within 96 hours. Overall, 46% of isolates were susceptible to cephalothin. Only 35% of coagulase-negative staphylococci were sensitive to cephalothin. No Gram-positive vancomycin resistance was encountered. CONCLUSION: Empiric prophylactic antibiotics for revision hip and knee arthroplasties should include vancomycin to cover Gram positive organisms and gentamicin for most Gram-negative bacteria. PMID- 19126903 TI - Approach-related complications of open versus thoracoscopic anterior exposures of the thoracic spine. AB - This article reviews the approach-related complications of open versus thoracoscopic anterior exposures of the thoracic spine and suggests possible ways to avoid them. PMID- 19126904 TI - Kinematic evaluation of the spine: a kinetic magnetic resonance imaging study. AB - Kinetic magnetic resonance images (kMRIs) of 587 lumbar and 459 cervical spines of symptomatic patients in axially loaded, upright neutral (0 degrees), flexion (40 degrees), and extension (-20 degrees) positions were evaluated. Imaging took 10 to 12 minutes to complete in each position. Cervical kinematics were significantly affected by intervertebral disc degeneration, cervical cord compression, and sagittal alignment of the cervical spine. kMRI was effective in diagnosing lumbar disc herniations that are often missed using conventional MRI. kMRI is effective for diagnosing, evaluating, and managing degenerative disease or injury within the spine. PMID- 19126905 TI - Reconstruction of large sacral defects following tumour resection: a report of two cases. AB - Sacral tumours often present surgical resection and reconstruction challenges. Wide resections result in large sacral defects and neoadjuvant radiotherapy impairs wound healing. The wounds need to be covered with bulky, well vascularised, healthy tissues. We present 2 cases where large sacral defects were reconstructed following tumour resection. Both defects were reconstructed with inferiorly based, transpelvic, pedicled vertical rectus abdominis myocutaneous flaps. This is a robust flap and carries a well-vascularised muscle bulk and skin paddle. The donor site is distant from the lesion site and is thus unaffected by both the resection and radiotherapy. This is a useful flap for reconstructing large sacral defects. PMID- 19126906 TI - Osteomyelitis variolosa: a report of two cases. AB - We present 2 patients with osteomyelitis variolosa in both elbow joints--sequelae of smallpox. The condyles were elongated and the central portions of the distal humeri were absorbed and the elbow joints were unstable. One of the patients sustained a closed fracture of the distal humerus. The fracture united uneventfully following stabilisation and bone grafting. At the 13-year follow-up, the patient had satisfactory elbow function. PMID- 19126907 TI - Spinal infection caused by Mycobacterium avium complex in a patient with no acquired immune deficiency syndrome: a case report. AB - Spinal infection caused by Mycobacterium avium complex (MAC) is rarely seen in people who do not have acquired immune deficiency syndrome. We report such a case in a 60-year-old man who underwent anterior spinal fusion after treatment with antibiotics had failed. The presentation of MAC spinal infection is different from that seen in MAC lung infection, with more than half presenting with urgent or semi-urgent neurological deficits. Younger patients who are not immunocompromised can also be infected. It should be considered as a differential diagnosis in patients with tuberculosis of the spine. The use of antibiotics should be based on the cultured organism's sensitivity results. Indications for surgery are progressive bony destruction, abscess formation, and neurological compression. PMID- 19126908 TI - Giant cell reparative granuloma of the proximal tibia: a case report. AB - Giant cell reparative granulomas (GCRGs) are non-neoplastic inflammatory lesions, usually of the jaw or gingiva or small bones of the hands and feet. We report one such case in the right proximal tibia of a 45-year-old man. Radiological studies showed a lytic lesion with marginal sclerosis in the epiphysis and metaphysis. After open biopsy, a preliminary diagnosis of a benign giant cell tumour was made. One month after admission, the lesion was curetted and filled with cancellous bone and hydroxyapatite. Based on the histology of the curetted lesion, the diagnosis was changed to a GCRG. The patient had an uneventful postoperative course, with no evidence of local recurrence and metastasis. He died from gastric cancer 2 years later. PMID- 19126909 TI - Primary muscular hydatidosis mimicking soft tissue tumour: a report of five cases. AB - Primary muscular hydatidosis is rare and usually presents as an asymptomatic, slowly growing mass mimicking a soft tissue tumour. Adequate preoperative planning and wide excision is recommended, as incomplete excision may lead to anaphylactic reactions and local recurrence. It should be considered in the differential diagnosis of soft tissue tumours especially in regions endemic for the parasite. PMID- 19126911 TI - Isolated rupture of the lateral collateral ligament during yoga practice: a case report. AB - We report a case of isolated rupture of the lateral collateral ligament (LCL) of the knee while attempting to place the left foot behind the head during yoga practice. The 34-year-old man had discomfort of the lateral aspect of the knee particularly with varus strain. A magnetic resonance image revealed rupture of the LCL at the insertion onto the fibula. The patient had grade-II laxity of the LCL and was treated non-operatively. At the 12-month follow-up, grade-I laxity of the LCL remained clinically evident, but function was not impaired. PMID- 19126912 TI - Postoperative fall after the use of the 3-in-1 femoral nerve block for knee surgery: a report of four cases. AB - We present a serious postoperative complication related to the use of femoral nerve block in 4 patients, each of whom fell and sustained further injury. Preoperatively, all patients underwent a 3-in-1 femoral nerve block with 30 to 35 ml of 0.25% levobupivacaine with 1:200,000 epinephrine, with guidance by a nerve stimulator. After the falls, neurological examination of the operated legs revealed reduced 2-point discrimination, pain, and/or light touch sensation. All patients underwent further operation for the fall injury and had delayed full weight bearing. We recommend that, after having a femoral nerve block, patients should undergo enhanced postoperative evaluation of blockade and proprioceptive function to ensure safe neurological function before mobilisation. PMID- 19126910 TI - The use of a reverse flow sural fasciocutaneous flap in a patient with multiple trauma: a case report. AB - Provision of soft-tissue coverage for defects in the distal leg and foot is a challenge, especially in patients with multiple injuries and major soft-tissue defects. Major flap reconstruction requires expertise and the results are variable, with high morbidity. We report a case in which a reverse flow sural fasciocutaneous flap was used for treatment of an open fracture-dislocation of the right ankle after repeated debridement in a 64-year-old man with a history of chronic smoking, diabetes mellitus, and hypertension. PMID- 19126913 TI - Intramedullary nailing for adult hypophosphatasia: a case report. AB - Hypophosphatasia is a rare genetic metabolic disorder characterised by defective bone mineralisation secondary to serum and bone alkaline phosphatase deficiency. We report a 46-year-old woman who underwent multiple intramedullary nailings for fractures and deformities of 6 long bones over 13 years. PMID- 19126914 TI - Piriformis pyomyositis: a report of three cases. AB - Pyomyositis is a subacute infection of skeletal muscles. It can be life threatening if diagnosis and treatment are delayed. Only 5 cases of isolated piriformis pyomyositis have been reported. We report signs and symptoms of piriformis pyomyositis in 3 women who were treated mainly with antibiotics. Computed tomography is useful in making the diagnosis. Early diagnosis and treatment may avoid surgical treatment and reduce mortality. PMID- 19126915 TI - Ossification of the thoracic ligamentum flavum in an achondroplastic patient: a case report. AB - We report a case of spinal stenosis with ossification of the thoracic ligamentum flavum in a 53-year-old man with achondroplasia. Neurological signs indicated flaccid paralysis below L1, and the patient was unable to walk. Ossification of the ligamentum flavum was observed at T4/5 and T9 to T12, compressing the thecal sac. Laminectomy of T9 to L1 was performed. At one-year follow-up, the patient was able to walk with one elbow crutch. PMID- 19126916 TI - Salvage arthrodesis for fracture-dislocation of the cuneonavicular and calcaneocuboid joints: a case report. AB - We present a 22-year-old man with dislocation of both the calcaneocuboid and cuneonavicular joints and fractures of the calcaneum and navicular of the right foot. The joints were reduced with percutaneous Kirschner wires, but the disrupted dorsal cuneonavicular ligaments were left unrepaired. Reduction was suboptimal and the joints were subluxed resulting in disabling arthralgia. Six months later, he underwent salvage arthrodesis of the subluxed calcaneocuboid and cuneonavicular joints. At 24-month follow-up, the patient had returned to work and remained pain-free when walking, with good fusion of both joints. Early anatomic reduction, stable fixation, and ligament reconstruction are essential for a good outcome. Arthrodesis is indicated when subluxation and posttraumatic arthritis are present. Primary arthrodesis is a viable option for severe midfoot fracture-dislocations, because it facilitates rehabilitation and functional recovery, and obviates the need for a secondary arthrodesis should arthritis arise. PMID- 19126917 TI - Laminoplasty for patients aged 75 years or older with cervical myelopathy. PMID- 19126918 TI - The use of qualitative cultures for detecting infection in open tibial fractures. PMID- 19126919 TI - The role of out-of-hospital cardiac arrest in predicting hospital mortality for percutaneous coronary interventions in the Clinical Outcomes Assessment Program. AB - Published mortality models for percutaneous coronary intervention (PCI), including the Clinical Outcomes Assessment Program (COAP) model, have not considered the effect of out-ofhospital cardiac arrest. The primary objective of this study was to determine if the inclusion of out-of-hospital cardiac arrest altered the COAP mortality model for PCI. The COAP PCI database contains extensive demographic, clinical, procedural and outcome information, including out-of-hospital cardiac arrest, which was added to the data collection form in 2006. This study included 15,586 consecutive PCIs performed in 31 Washington State hospitals in 2006. Using development and test sets, the existing COAP PCI logistic regression mortality model was examined to assess the effect of out-of hospital arrest on in-hospital mortality. Overall, 2% of individuals undergoing PCI had cardiac arrest prior to hospital arrival. Among 8 hospitals with PCI volumes < 120 cases per year, 4 had cardiac arrest volumes that exceeded 10% of total volume, whereas none of the centers with > 120 cases per year did. In hospital mortality was 19% in the arrest group and was 1.0% in remaining procedures (p < 0.0001). In the new multivariate model, out-of-hospital cardiac arrest was highly associated with mortality (odds ratio = 5.50; 95% confidence interval [CI] = 3.28-9.25). When evaluated in the test set, the new model had excellent discrimination (c-statistic = 0.89; 95% CI = 0.85-0.93). Out-of hospital cardiac arrest is an important determinant of risk-adjusted in-hospital mortality for PCI, particularly for hospitals with low volumes and relatively high volumes of cardiac arrest cases. PMID- 19126920 TI - Analysis of particulate debris after superficial femoral artery atherectomy. AB - PURPOSE: To evaluate the occurrence, size and composition of embolized debris captured during routine directional atherectomy using the SilverHawk device. METHODS: 15 consecutive eligible patients with a nonocclusive superficial femoral artery (SFA) were enrolled. Patients were included if they were > 18 years of age and had > or = 70% stenosis in the SFA. All lesions underwent plaque excision with the SilverHawk atherectomy device. A FilterWire EZ was used for distal protection and retrieval of embolized material. Specimens were collected separately from the filter basket and the SilverHawk atherectomy device's nosecone and were studied by a pathologist for number, size and composition. RESULTS: Visible debris captured in the filter was found in the majority of patients 14/15 (93%). Clinically-significant debris was found in 7/15 (47%) patients. The proportion of captured debris ranged from 0.1-0.4 cm. Microscopy revealed that the shaved particles consisted predominantly of collagen, fibrin, lipid-laden macrophages, cholesterol and calcium. Analysis of the embolized material revealed a different composition, mostly consisting of collagen with fibrosis, cholesterol and macrophages. CONCLUSION: In this single-center comparative study we have shown that during SilverHawk atherectomy of SFA lesions, distal embolization is universal. The debris captured in the filter is different in overall composition from the captured material in the nosecone of the SilverHawk device. Debris large enough to cause clinically-significant embolization, no-reflow and ischemia following SFA interventions occurred in nearly 50% of cases. PMID- 19126921 TI - Plaque excision with distal protection: a logical next step for superficial femoral artery SilverHawk atherectomy. PMID- 19126922 TI - Level of complement activity predicts cardiac dysfunction after acute myocardial infarction treated with primary percutaneous coronary intervention. AB - BACKGROUND: The positive effect of reperfusion after ST-elevation myocardial infarction (STEMI) can be reduced by ischemic/reperfusion (I/R) injury.Mannose binding-lectin (MBL) and soluble C5b-9 (membrane-attack-complex) are involved in complement-driven cell lysis and may play a role in human myocardial I/R injury. We evaluated the potential association between MBL and sC5b-9 in plasma and subsequent cardiac dysfunction in patients with STEMI treated with primary percutaneous coronary intervention (pPCI). METHODS: The study included 74 STEMI patients with acute occlusion of the left anterior descending coronary artery who were successfully treated with pPCI. Cardiac dysfunction was defined as left ventricular ejection fraction LVEF < 35%. RESULTS: Patients with subsequent LVEF < 35% had significantly higher median MBL and lower sC5b-9 compared to patients with LVEF > or = 35%. After adjustment of the multivariate logistic regression analysis, the odds for reduced LVEF was 5.5 (95% CI:1.5-19.3; p = 0.01) for patients with MBL > or = 800 mcg/L, and 5.0 (95% CI 1.4-18.4; p = 0.01) for patients with sC5b-9 < or = 160 mcg/L. CONCLUSION: High plasma MBL and low plasma sC5b-9 are independently associated with increased risk of cardiac dysfunction in STEMI patients treated with pPCI, probably due to increased complement activity during the ischemic and reperfusion process. The predictive value of low peripheral plasma sC5b-9 may be explained by an accumulation and activation of sC5b-9 in the infarcted myocardium. PMID- 19126923 TI - Impact of body mass index on long-term all-cause mortality after percutaneous coronary intervention in African-Americans. AB - BACKGROUND: Studies on body mass index (BMI) and post-percutaneous coronary intervention (PCI) outcomes have suggested an "obesity paradox" (lower post-PCI mortality rates in obese patients compared to patients of normal weight). Hypothesis. We assessed the hypothesis that BMI is an independent predictor of post-PCI long-term mortality in African-Americans. METHODS: We evaluated 777 patients (146 with normal BMI [BMI > 18.5 to < 25 kg/m2], 261 overweight patients [BMI > or = 25 to < 30 kg/m2], and 370 obese patients [BMI > or = 30 kg/m2]) who underwent PCI during January 2003 to August 2006. RESULTS: After a median follow up of 4 +/- 1 years, the overall mortality rate was 10.5% (82 deaths). The survival rate was 84%, 90% and 92% in the normal BMI, overweight and obese groups, respectively (p = 0.014 by log-rank test; hazard ratio relative to the obese group was 2.2 for the normal weight and 1.2 for the overweight groups). After adjustment for baseline clinical and procedural characteristics (age, smoking history, chronic renal insufficiency, end-stage renal disease, left main coronary artery intervention, preprocedure hemoglobin, left ventricular ejection fraction and successful PCI) using a Cox proportional hazards model, there was no significant difference in the long-term all-cause mortality rate among the three groups (p = 0.93). CONCLUSION: BMI does not appear to be an independent predictor of long-term mortality after PCI in African-Americans. PMID- 19126924 TI - Hyperperfusion syndrome following carotid artery stenting: the largest single operator series to date. AB - BACKGROUND: Cerebral hyperperfusion syndrome (HPS) results from autoregulatory failure of cerebral blood flow following carotid endarterectomy (CEA) or carotid artery stenting (CAS) and encompasses a range of neurological findings including headache, seizure, intracranial hemorrhage (ICH), altered mental status and focal neurological changes. This report is the largest single-operator series evaluating the incidence and predictors of HPS following CAS. METHODS: A retrospective review was conducted on 482 consecutive patients who underwent CAS between August 1999 and December 2007 at Baptist Medical Center--Princeton, Birmingham, Alabama. All interventions were performed by a single operator (FM). The mean patient age was 70.4 +/- 10.3 years and 36% were symptomatic. All patients were high-risk for CEA. After cerebral protection catheters were routinely available, they were used in all but 6 cases (98.1%) where the anatomy precluded delivery. Brain computed tomography (CT) was performed immediately for any neurological change or significant headache following CAS. After neurological consultation and imaging, HPS was diagnosed if: 1) a neurological change occurred (not simply a headache); 2) CT revealed ipsilateral sulcal effacement/cerebral edema; and 3) stroke or transient ischemic attack (TIA) was excluded. RESULTS: Seven patients (1.45%) developed HPS following CAS. All patients achieved complete neurological recovery 6-24 hours following the procedure. Patients who developed HPS were significantly more likely to have had recent transient ischemic attack (TIA) symptoms than patients without HPS (p = 0.04). Unlike previous reports, there were no significant differences in procedural details, lesion characteristics and post-procedure blood pressure between the HPS and non HPS patients, although the number of cases was small. Overall, the HPS cohort had a higher prevalence of comorbidities, though these differences did not reach statistical significance. Hypertension was present in all 7 HPS patients. Other complications in the series were death (0.83%), stroke (1.87%) and TIA (1.45%). CONCLUSIONS: The incidence of HPS is low (1.45%) following CAS, but it is an important complication to distinguish from stroke and TIA. Patients with a recent TIA may be predisposed to HPS. This report may underestimate the incidence of HPS, since patients with an isolated headache did not meet our diagnostic criteria and routine post-procedure brain CT imaging was not performed. The clinical predictors of HPS and its optimum management remain to be determined. PMID- 19126925 TI - Intravascular ultrasound evaluation of a stenotic modified Blalock-Taussig. PMID- 19126926 TI - Severe myocardial ischemia due to a large vascular malformation of the left anterior descending coronary artery with a fistulous connection from the right coronary artery. PMID- 19126927 TI - Acute anterior myocardial infarction secondary to a myocardial muscular bridge. PMID- 19126928 TI - Post cardiac injury syndrome after uncomplicated coronary angioplasty. PMID- 19126929 TI - Acute myocardial infarction in two young women without significant risk factors. AB - Spontaneous coronary artery dissection is an uncommon cause of acute coronary syndrome in young healthy females, with pregnancy and the use of oral contraceptive pills as recognized predisposing factors. We present two cases of acute myocardial infarction in two young females without significant risk factors and review the literature on spontaneous coronary artery dissection including diagnosis and treatment options. PMID- 19126930 TI - Endovascular treatment of traumatic bilateral internal carotid artery dissection. PMID- 19126931 TI - Left main coronary artery stenosis following surgical valve replacement: changing valvular into ischemic heart disease. AB - Left main stem coronary artery (LMSCA) stenosis after surgical aortic valve replacement (AVR) is a rare but potentially lethal complication. Among the different possible causes, antegrade cardioplegia for direct coronary perfusion seems to be the most significant and prevalent. Here we discuss two cases of late presentation of iatrogenic coronary ostial stenosis following valve replacement surgery. Both cases were successfully treated with percutaneous intervention and stent implantation. PMID- 19126932 TI - "Hit by thrombotic events" following heparin--think of HIT. PMID- 19126933 TI - Phosphodiesterase type 5 inhibitors in pulmonary hypertension: a Hobson's choice for Indian patients. PMID- 19126934 TI - Primary angioplasty vs. thrombolysis. AB - Several randomized trials and meta-analyses have shown that primary angioplasty is superior to thrombolysis in the treatment of ST-segment elevation myocardial infarction (STEMI) in terms of death, re-infarction and stroke. However, primary angioplasty should be regarded as the preferred strategy unless it could not be applied with a reasonable time-delay to treatment as compared to the administration of thrombolysis. In fact, time-to-treatment has shown to be a determinant of survival not only for thrombolysis but also for primary angioplasty. Recent guidelines consider a time of 90 minutes from first medical contact to Percutaneous Coronary Intervention (PCI) or a PCI-related time delay of 60 minutes as reasonable cut-offs to identify the best reperfusion strategy. The beneficial effects of primary angioplasty could be expected particularly after the first 3 hours from symptoms onset when thrombolysis, particularly streptokinase, may be less effective, whereas within the first 3 hours, thrombolysis (started in the pre-hospital setting, preferably) may represent a valid therapeutic option. Since the survival benefits of primary angioplasty depends on patient's risk profile and timely application of reperfusion, we would suggest, among patients in the first hours from symptoms onset, a strategy of early pharmacological reperfusion and transfer to primary PCI centers where the decision of performing angiography acutely may be based on the assessment of myocardial reperfusion and risk profile, whereas after the first 3 hours from symptoms onset, primary angioplasty should be considered as the preferred strategy, if applicable, particularly in regions when streptokinase still represents the only available lytic therapy. PMID- 19126935 TI - Stroke prevention in atrial fibrillation--pharmacologic strategies. AB - Stroke prevention in atrial fibrillation presents a challenge for many clinicians. A considerable amount of research has gone into creating scoring systems that can accurately predict thromboembolic risk. Patients at low enough risk are best treated with aspirin, whereas higher-risk patients require anticoagulation with warfarin despite its shortcomings. Antithrombotic regimens for moderate-risk patients require careful consideration based on bleeding risk, age, and the ability to undergo frequent serum monitoring. Oral drugs may soon be available to replace warfarin. While this may simplify dosing and monitoring, it will still be necessary to weigh the risks and benefits of therapy. PMID- 19126936 TI - An observational study to know the association of leukocytosis and fever with infection in post cardiac surgery patients. AB - BACKGROUND: Leukocytosis and fever in the absence of infection are common phenomena in post surgical period. The systemic inflammatory response syndrome induced by surgical trauma is a well-known entity, with resultant release of variety of inflammatory cytokines leading to fever and/or leukocytosis in spite of the absence of infection. AIMS AND OBJECTIVE: To know the association of leukocytosis and fever with infection in immediate post cardiac surgical period. METHODS: It was a retrospective, observational study including 569 patients, who underwent cardiac surgeries in our institute. All demographic data, comorbidities and other factors associated with fever and/or leukocytosis were analyzed. The sensitivity and specificity of fever and/or leukocytosis were analyzed for the diagnosis of infection. RESULTS: Out of 569 patients; 49 patients (9%) had evidence of infection on microbiological culture. Mean total leukocyte counts (TLC) on zero and 1st postoperative days were associated with infection with a sensitivity (75.5% and 63.3%) and specificity (15.8% and 24.0%) respectively. Mean maximum temperature (Tmax) on zero and 1st postoperative days were associated with infection with a sensitivity (20.4% and 24.5%) and specificity of (82.5% and 83.3%) respectively. The combined sensitivity and specificity of leukocytosis and fever for the diagnosis of infection on 2nd postoperative day was 14.3% and 91.5% respectively. CONCLUSION: Fever and leukocytosis are poor predictors of diagnosing infection on the first two postoperative days. However, fever and leukocytosis combined together have low sensitivity (14.3%) with high specificity (91.5%) for the diagnosis of infection on the 2nd and subsequent postoperative days. PMID- 19126937 TI - Efficacy and safety of PDE-5 inhibitor tadalafil in pulmonary arterial hypertension. AB - BACKGROUND: Pulmonary arterial hypertension (PAH) is a life threatening disease for which phosphodiesterase-5 inhibitor sildenafil is recently approved. We aimed to evaluate the efficacy and safety of tadalafil, a long acting congener of sildenafil, in treatment of PAH related to previous left to right shunt lesions. METHODS: In this blinded, cross over study, 11 patients with severe PAH related to congenital left to right shunt lesions (Eisenmenger syndrome) were randomly assigned to tadalafil (20 mg daily) or placebo for 4 weeks period, separated by a wash out period of at least 2 weeks. They were symptomatic with a six minute walk distance (6MWD)>or=50 m. The change in 6MWD, echo-Doppler determined pulmonary artery systolic pressure (PASP), WHO Class and modified Borg Dyspnea Index (BDI) were assessed after each therapy. RESULTS: Eight patients who completed the study protocol were analyzed. Tadalafil was associated with a significant increase in 6MWD (mean 409.25 SD 40.25 m vs 319.37 SD 42.39 m, p<0.0001), reduction in PASP (88.75 SD 23.26 mmHg vs 109.5 SD 23.78 mmHg, p<0.0001), improvement in BDI (4.62 SD 2.56 vs 6.37 SD 2.61, p=0.021) and WHO Class (6 patients vs 2 patients), compared to placebo. Tadalafil was well tolerated with no significant untoward effects. CONCLUSIONS: Tadalafil, in patients with PAH related to previous congenital left to right shunt lesions, lead to a significant improvement in exercise capacity (6MWD), PASP and WHO Class with reduced perceived exertion and was well tolerated. PMID- 19126938 TI - Oral tadalafil in pulmonary artery hypertension: a prospective study. AB - BACKGROUND: Pulmonary artery hypertension is a disorder with limited treatment options and poor prognosis. We studied the effect of tadalafil, a selective phosphodiesterase type 5 inhibitor, in patients with this disease. METHODS AND RESULTS: In thirteen patients (5 males, 8 females) aged 15-70 years, with pulmonary artery hypertension (7 with congenital heart disease, 4 with chronic pulmonary thromboembolism and 2 with primary pulmonary hypertension), tadalafil was added to their baseline therapy at 10 mg OD for two weeks and if tolerated well, increased to 20 mg OD for the next two weeks. After four weeks of therapy, all thirteen patients had subjective improvement in the form of overall well being and objective improvement as measured by duration of exercise on treadmill following Bruce protocol from a baseline of 350.54+/-255.06 seconds to 479.54+/ 195.00 seconds (p<0.01). There was improvement in NYHA functional class in eleven patients. Two patients, though showed no improvement in functional class, had subjective improvement in effort tolerance and objective improvement as measured by duration of exercise on TMT. Hemodynamic parameters showed a trend towards improvement, mean pulmonary artery mean pressure decreased from 63.5+/-26.2 mmHg to 62.2+/-24.8 mmHg (p>0.05), mean pulmonary blood flow increased from 3.26+/ 1.04 L/min to 3.44+/-1.26 L/min (p>0.05), mean total pulmonary vascular resistance decreased from 1858.6+/-1138.9 dyne-sec.cm-5 to 1737.3+/-1017.2 dyne sec.cm-5 (p>0.05). CONCLUSION: Oral tadalafil was well tolerated and had a beneficial effect in patients with pulmonary artery hypertension irrespective of age, sex and underlying etiology. Patient had improved effort tolerance and a trend towards improved pulmonary hemodynamics. PMID- 19126939 TI - Sildenafil improves survival in idiopathic pulmonary arterial hypertension. AB - AIMS AND OBJECTIVES: Idiopathic Pulmonary Arterial Hypertension (IPAH) is a serious disorder of unknown etiology with limited therapeutic options. Sildenafil has been shown to decrease symptoms, improve hemodynamics and quality of life. Its impact on survival is uncertain. We studied the efficacy of sildenafil in improving survival in patients with IPAH. METHODS AND RESULTS: Data on survival of patients with IPAH was collected from prospectively maintained registry at our hospital from January 1999 to December 2005. Thirty nine patients who were treated with conventional therapy including digoxin, diuretics, anticoagulants and calcium channel blockers prior to January 2001 served as historical controls (control group). One hundred and thirty nine patients received sildenafil additionally from January 2001 (sildenafil group). All patients in sildenafil group showed improvement in symptoms. Survival of patients in sildenafil group was significantly better compared to historical controls receiving only conventional therapy. It was 89%, 43% and 19% in the control group Vs 93%, 75% and 54% in the sildenafil group at the end of 1, 3 and 5 years respectively (P Value=0.0002). Sildenafil was well tolerated and none of the patients had to discontinue the treatment. CONCLUSION: Sildenafil when added to conventional therapy improves symptoms as well as survival significantly compared to conventional therapy alone. Further randomized controlled trials are needed to evaluate its impact on survival when used either alone or in combination with other drugs. PMID- 19126940 TI - Sildenafil for pulmonary hypertension secondary to congenital heart diseases. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of oral Sildenafil for treatment of pulmonary hypertension secondary to congenital heart diseases. METHODS & RESULTS: Twelve patients with un-repaired congenital heart diseases resulting in significant pulmonary hypertension were enrolled in a prospective trial of oral Sildenafil treatment for 12 weeks. The outcomes measured were change in the systemic oxygen saturations, 6-minute walk test distance, and New York Heart Association class. The systemic blood pressure and visual symptoms were monitored for evaluating the tolerability of Sildenafil. The mean age was 25.7+/-14.4 years. The mean pulmonary vascular resistance before treatment was 17.4 Wood units. After 12 weeks of sildenafil, an increase in oxygen saturations (87.8+/-5.3%-90.25+/-4%, p=0.04) with an associated increase of one New York Heart Association class (p=0.009) was noted. A non-statistically significant increase in the 6-minute walk test distance (310.33+/-42-333.75+/ 54.6 meters, p=0.47) was also noted. Sildenafil was well-tolerated with no significant change in systemic blood pressure or visual side-effects. CONCLUSION: Oral Sildenafil is well-tolerated and produces some improvement in the oxygenation and functional status of patients with pulmonary hypertension secondary to congenital heart diseases. PMID- 19126941 TI - Trends in prevalence of coronary risk factors in an urban Indian population: Jaipur Heart Watch-4. AB - BACKGROUND: To determine prevalence of multiple coronary risk factors in a North Indian Punjabi community and to compare these with previous population based studies in the same city in North India we performed an epidemiological study. METHODS: A community-based epidemiological study that focused on lifestyle determinants of obesity and its correlates in migrants from Punjab was performed at a single location in Jaipur. A house-to-house enumeration was performed to enroll all adults>or=20 years age in the locality who were then invited for participation in the study. Of the 1400 eligible subjects, 1127 participated (response rate 80.5%, men 556, women 571). Risk factor measurements included smoking or tobacco use, body-mass index (BMI), waist:hip ratio (WHR) and body fat, and in 644 (56.6%) subjects (men 340, women 304) blood examination for fasting blood glucose and lipids. Coronary risk factors were determined using pre specified criteria. RESULTS: There was a significant prevalence of risk factors in both men and women respectively with smoking or tobacco use in 209 (37.6%) and 12 (2.2%), obesity (BMI>or=25 kg/m2) in 303 (54.5%) and 350 (61.3%), truncal obesity (high WHR) in 339 (61.0%) and 310 (54.30%), hypertension in 322 (57.9%) and 279 (48.9%), high total cholesterol>or=200 mg/dl in 111 (32.6%) and 120 (39.5%), low HDL cholesterol<40 mg/dl in 103 (30.3%) and 83 (27.3%), high triglycerides>or=150 mg/dl in 146 (42.9%) and 132 (43.4%), metabolic syndrome in 166 (48.8%) and 137 (45.1%), and diabetes in 88 (25.9%) and 64 (21.1%) subjects. In both men and women there was a significant age-associated escalation in obesity, central obesity, hypertension, high cholesterol and diabetes prevalence (Mantel-Haenszel chi2 for trend p<0.05). Logistic regression analyses revealed that obesity and truncal obesity were major determinants of multiple risk factors such as hypertension, hypercholesterolemia, metabolic syndrome and diabetes (age adjusted odds ratios p<0.01). Comparison with previous population-based risk factor studies from the same city in years 1995 and 2002 revealed that risk factors were significantly greater in the present group. Age-stratified differences revealed that obesity at younger age was more frequent in the present cohort. CONCLUSIONS: There is a significant prevalence of multiple cardiovascular risk factors in this population group. Obesity is a major determinant of multiple risk factors and appears at a younger age compared to other studies in the same location. PMID- 19126942 TI - Heparin induced thrombocytopenia management with bivalirudin. AB - We report a case of Heparin Induced Thrombocytopenia (HIT) following percutaneous coronary intervention. The case is unique in that thrombocytopenia occurred very early after heparin administration and responded well to a regime of bivalirudin a direct thrombin inhibitor readily available in India. Heparin, Thrombocytopenia, Bivalirudin Acute HIT, occurring within few hours of heparin therapy have been reported in patients previously exposed to unfractionated heparin (UFH) or low molecular weight heparin (LMWH) 1. Prompt recognition of the condition and timely intervention with direct thrombin inhibitors, can result in salvaging patients from this potentially fatal complication. PMID- 19126943 TI - Leiomyoma in the heart. PMID- 19126944 TI - Electrocardiographic and echocardiographic findings during the recent outbreak of viral fever in National Capital Region. PMID- 19126945 TI - Transcatheter coil occlusion of IMA graft side branches for relief of coronary steal. PMID- 19126946 TI - Prominent isovolumic relaxation velocities in mitral and tricuspid inflows in cardiac tamponade: normalisation after pericardiocentasis. PMID- 19126947 TI - Metabolic modulation as a target for heart failure treatment: current perspective. PMID- 19126948 TI - Prevention of contrast induced nephropathy--an update. PMID- 19126949 TI - Non-cardiac surgery in patients with drug eluting stents: considerations on antiplatelet therapy. PMID- 19126950 TI - Coil embolization of septal artery as a treatment for hypertrophic obstructive cardiomyopathy. PMID- 19126951 TI - Delayed response following trans-catheter coil embolization in HOCM. PMID- 19126952 TI - Iatrogenic left main coronary artery stenosis. PMID- 19126953 TI - Multislice CT for femoral pseudoaneurysm after angioplasty. PMID- 19126957 TI - Targeted imaging of neoplasia in the digestive tract. PMID- 19126958 TI - In vivo imaging of cancer biomarkers using activatable molecular probes. AB - With the continued advancements in cellular and molecular biology, especially in the areas of genomics, proteomics and metabolomics, the scientific research community is gaining ground in uncovering the tortuous details associated with cancer. Molecular imaging has undergone a similar progression and is showing promise as a future method to aid in the early detection of malignancies, locating metastatic disease, staging tumors, evaluating the availability of therapeutic targets, and monitoring the efficacy of treatment. A subset of molecular imaging contrast agents known as "activatable molecular probes" has generated a particularly high level of excitement in the imaging community. Activatable molecular probes are designed to elicit a detectable change in signal upon enzymatic activity or in response to specific biomolecular interactions. In many cases, these unique characteristics allow for very high signal-to-background ratios compared with conventional targeted contrast agents and they open up the possibility of imaging intracellular targets. In this review, we will discuss some of the activatable probes recently developed for optical and magnetic resonance imaging platforms and their use in the visualization of cancer biomarkers in vivo. PMID- 19126959 TI - Quantum dot bioconjugates for in vitro diagnostics & in vivo imaging. AB - Semiconductor quantum dots are tiny light-emitting nanocrystals (2-10 nm) that have captivated researchers in the biomedical field in the last decade. Compared to organic dyes and fluorescent proteins, quantum dots (QDs) have unique optical properties such as tunable emission spectra, improved brightness, superior photostability, and simultaneous excitation of multiple fluorescence colors. Since the first successful reports on biological use of QDs a decade ago, QDs and their bioconjugates have been successfully applied in various imaging applications including fixed cell labeling, imaging of live cell dynamics, in situ tissue profiling, fluorescence detection, sensing and in vivo animal imaging. In this review, we will cover the optical properties of QDs, the biofunctionization strategies, their in vitro diagnostic applications and in vivo imaging applications. In addition, we will discuss the making of a new class of QDs--the self-illuminating QDs and their in vivo imaging and sensing applications. We will conclude with the issues and perspectives on QDs as in vivo imaging probes. PMID- 19126960 TI - Labeled anti-mucin antibody detectable by infrared-fluorescence endoscopy. AB - The goal of our study was to develop a method for molecular imaging of the gastrointestinal tract using an infrared fluorescence endoscope (IRFE) and antibodies labeled with an indocyanine green (ICG) derivative to detect cancerous tissue. The IRFE comprised an infrared endoscope equipped with excitation (710 790 nm) and barrier (810-920 nm) filters. We developed ICG-N hydroxysulfosuccinimide ester (ICG-sulfo-OSu) and 3-ICG-acyl-1,3-thiazolidine-2 thione (ICG-ATT) as infrared fluorescent-labeling reagents, and anti-human carcinoembryonic antigen (CEA) antibody and MUC1 antibody were labeled with the ICG-derivatives. Freshly resected specimens of gastric cancer were observed by IRFE after reaction with ICG-derivative-labeled antibodies. Positive fluorescence was observed at the tumor location by IRFE, and the immunofluorescent images correlated well with the tumor sites. The immunofluorescence studies suggested that the intensity of the infrared fluorescence of the ICG-ATT-labeled MUC1 antibody is stronger than the ICG-sulfo-OSu-labeled MUC1 antibody. We concluded that specific antibodies for gastrointestinal cancer labeled with an ICG derivative accompanied by a reinforcing agent and an optimal electronic device can generate a strong enough fluorescent signal to visualize cancer proteins. PMID- 19126961 TI - In vivo biomarkers for targeting colorectal neoplasms. AB - Colorectal carcinoma continues to be a leading cause of cancer morbidity and mortality despite widespread adoption of screening methods. Targeted detection and therapy using recent advances in our knowledge of in vivo cancer biomarkers promise to significantly improve methods for early detection, risk stratification, and therapeutic intervention. The behavior of molecular targets in transformed tissues is being comprehensively assessed using new techniques of gene expression profiling and high throughput analyses. The identification of promising targets is stimulating the development of novel molecular probes, including significant progress in the field of activatable and peptide probes. These probes are being evaluated in small animal models of colorectal neoplasia and recently in the clinic. Furthermore, innovations in optical imaging instrumentation are resulting in the scaling down of size for endoscope compatibility. Advances in target identification, probe development, and novel instruments are progressing rapidly, and the integration of these technologies has a promising future in molecular medicine. PMID- 19126963 TI - Disease related glycosylation changes and biomarker discovery: challenges and possibilities in an emerging field. Editorial. PMID- 19126962 TI - In vivo cancer biomarkers of esophageal neoplasia. AB - The emergence of in vivo cancer biomarkers is promising tool for early detection, risk stratification, and therapeutic intervention in the esophagus, where adenocarcinoma is increasing at a rate that is faster than any other in industrialized nations. Exciting advances in target identification, probe development, and optical instrumentation are creating tremendous new opportunities for advancing techniques of molecular imaging. Progress in these areas is being made with small animal models of esophageal cancer using surgical approaches to induce reflux of acid and bile, and these findings are beginning to be evaluated in the clinic. Further identification of relevant targets, characterization of specific probes, and development of endoscopic imaging technologies are needed to further this direction in the field of molecular medicine. In the future, new methods that use in vivo cancer biomarkers for the early detection of neoplastic changes in the setting of Barrett's esophagus will become available. PMID- 19126964 TI - Glyco-biomarkers: potential determinants of cellular physiology and pathology. AB - Once dismissed as just the icing on the cake, sugar molecules are emerging as vital components in life's intricate machinery. Our understanding of their function within the context of the proteins and lipids to which they are attached has matured rapidly, and with it the far reaching clinical implications are becoming understood. Recent advances in high-throughput glycomic techniques, glyco biomarker profiling, glyco-bioinformatics and development of increasingly sophisticated glyco-arrays, combined with our increased understanding of the molecular details of glycosylation have facilitated the linkage between aberrant glycosylation and human diseases, and highlighted the possibility of using glyco biomarkers as potential determinants of disease and its progression. The focus of this review is to give an insight into the biological significance of these glycomodifications, highlight some specific examples of glyco-biomarkers in relation to autoimmunity and in particular rheumatoid arthritis, and to explore the exciting possibility of exploiting these for diagnostic and prognostic strategies. PMID- 19126965 TI - Altered glycosylation in tumours focused to cancer diagnosis. AB - The lack of specific and sensitive tumour markers for early detection of cancer is driving a search for new approaches that could identify biomarkers. Markers are needed to alert clinicians at the early stages of tumourogenesis, before the cancer has metastasized, when the therapeutic drugs are more effective. Most tumour markers currently used in clinics are serum glycoproteins, frequently highly glycosylated mucins. Typically, the disease marker is the protein and not the glycan moiety of the corresponding glycoprotein or mucin. The increasing knowledge of the role of glycans in cancer suggests that further studies may assist both in determining their role in every step of tumour progression, and in the design of new therapeutic and diagnosic approaches. Detection of the altered glycans in serum tumour glycoproteins could be a way to achieve specificity in tumour detection. In this review, we focus on the glycan changes of two serum glycoproteins, prostate specific antigen--currently used as a tumour marker of prostate cancer--and human pancreatic ribonuclease in pancreatic adenocarcinoma. The detection of glycan changes, associated with subsets of glycoforms in serum glycoproteins that are specific to the tumour situation, could be the basis for developing more specific biomarkers. PMID- 19126966 TI - Glycosylation changes on serum glycoproteins in ovarian cancer may contribute to disease pathogenesis. AB - Ovarian cancer is the most lethal of all gynaecological cancers among women. Serum CA125 is the only biomarker that is used routinely and there is a need for further complementary biomarkers both in terms of sensitivity and specificity. N glycosylation changes in ovarian cancer serum glycoproteins include a decrease in galactosylation of IgG and an increase in sialyl Lewis X (SLe(x)) on haptoglobin beta-chain, alpha1-acid glycoprotein and alpha1-antichymotrypsin. These changes are also present in chronic inflammation but not in malignant melanoma, where there are low levels of inflammatory processes. Acute phase proteins carrying increased amounts of SLe(x) have an increased half-life. Sialylation of acute phase proteins also decreases apoptosis favouring survival of cancer cells. Cancer cells produce inflammatory cytokines which influence glycosylation processing in liver parenchymal cells. Altered glycosylation of the acute phase protein transferrin plays an important role in iron homeostasis. Glycosylated transferrin and its glycans have anti-apoptotic properties and many transferrin receptors in carcinoma could play a role in development of anaemia. Decreased galactosylation and sialylation of IgG increases the cytotoxicity of natural killer cells and complement activation via mannose-binding lectin (MBL). Altered glycosylation of acute phase proteins and IgG suggests that cancer regulates certain pathways favouring cancer cells survival. PMID- 19126967 TI - Carbohydrate markers in colon carcinoma. AB - Spontaneously mutated multiple oncogenes and/or tumor suppressor genes in colon epithelial cell and its progeny, may cause proliferation out of control and create benign colon neoplasm (colon polyp). If additional mutations involve genes responsible for cell adhesion and movement, aberrant epithelial cells may become malignant (colon cancer) and invade surrounding and remote tissues, creating secondary tumors called metastases. Incidence of colorectal cancer dramatically increases at 50-65 year of age. In Europe in 2006 colorectal cancer consisted 12.9% of all cancers and caused 207,400 deaths. To laboratory detection and monitoring of colon cancer are used tumor markers. Tumor markers are substances produced by the body in response to cancer, or by cancer tissue itself. Glycoconjugate markers for colon cancer include aberrant: mucins covering the surface of the colon epithelial cells, cadherins, selectins and Ig-like adhesion molecules mediating cell-cell adhesion, integrins and integral membrane proteoglycans responsible for adhesion of colon epithelial cells to extracellular matrix, glycoconjugate components of ECM, as well as lysosomal membrane glycoproteins and exoglycosidases. Detection of colon cancer at early non malignant stage is crucial in its prevention and eradication. As colon cancer is the effect of accumulation many somatic mutations in oncogens, supressors, mismatch repair genes and many genes responsible for posttranslational modifications of proteins, multidirectional approach should be applied for its detection. A glycobiological approach to diagnosis and treatment of colorectal cancer should be directed to detection changes in glycosylation accompanying every step of colon cancer progression, and correlation between changes in glycosylation and tumor progression. PMID- 19126968 TI - Glycomic approach for potential biomarkers on prostate cancer: profiling of N linked glycans in human sera and pRNS cell lines. AB - Prostate cancer is a leading cause of cancer death among men. Currently available screening test measures prostate-specific antigen (PSA) to detect prostate cancer. However, this test produces false positive values that often lead to negative biopsies. Therefore, a more reliable diagnostic tool is needed. Glycans in serum are of particular interest as around half of all proteins are glycosylated. In this study, N-linked glycans were enzymatically released by PNGase F from prostate epithelial cell lines (pRNS) expressing wild type or mutant androgen receptors and a small set of human serum samples. Released glycans were purified and partitioned into neutral and acidic components by solid phase extraction (SPE) using graphitized carbon cartridges. The SPE fractions were analyzed by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FT-ICR MS). Significant changes in some high-mannose and fucosylated biantennary complex N-linked glycans were observed in the serum of prostate cancer patients. PMID- 19126969 TI - Fucosylated glycoproteins as markers of liver disease. AB - Changes in N-linked glycosylation are known to occur during the development of various diseases. For example, increased branching of oligosaccharides has been associated with cancer metastasis and has been correlated to tumor progression in human cancers of the breast, colon and melanomas. Increases in core fucosylation have also been associated with the development of hepatocellular carcinoma (HCC). Recently, changes in both the total serum glycome and the glycosylation of specific IgG molecules have been observed in people with liver fibrosis and cirrhosis. The mechanisms by which changes in glycosylation are observed and their use as biomarkers of disease will be discussed. PMID- 19126970 TI - Glycosylation of serum proteins in inflammatory diseases. AB - Inflammatory diseases are accompanied by numerous changes at the site of inflammation as well as many systemic physiological and biochemical changes. In the past two decades more and more attention is being paid to changes in glycosylation and in this review we describe some of the changes found on main serum proteins (alpha1-acid glycoprotein, immunoglobulin G, immunoglobulin A, transferrin, haptoglobin, alpha2-macroglobulin, C-reactive protein, and others). Molecular background and physiological importance of most of these changes are yet to be discovered, but it is evident that glycosylation plays an important role in the inflammatory response. Maybe the greatest value of these changes currently lays in their potential diagnostic and prognostic usage, either in combination with current diagnostic markers or on their own. However, determining glycan structures is still technically too complex for most clinical laboratories and further efforts have to be made to develop simple analytical tools to study changes in glycosylation. PMID- 19126971 TI - Trachea rupture lesion in children: the two extremes. AB - Neck trauma in childhood resulting in trachea rupture lesions is an extremely rare but always life-threatening injury. Even if initial clinical signs are minimal, significant airway compromise always has to be considered. Mechanisms of accident and etiologies regarding especially the true impact forces of the trauma are various making standardized emergency management and health care difficult. Two opposite and exceptional case reports are presented and discussed in front of an extended literature overview. PMID- 19126972 TI - A novel system using the Forced Oscillations Technique for the biomechanical analysis of swallowing. AB - Physiological studies of swallowing and the diagnosis and treatment of dysphagia are crucially dependent of detailed information of respiratory and feeding events. However, this information has been obtained by indirect and/or expensive methods, as well as by methods demanding exposure to radiation. In this context, the purpose of this study was twofold: (1) describe a new low-cost system for the analysis of the respiratory events during swallowing using the Forced Oscillation Technique and (2) evaluate the performance of this device in the description of physiological events during water swallowing. The device consists of a personal computer adapted to measurement modules able to characterize the soft palate movement, the elevation of the larynx, the duration of deglutition apnoea, and the direction of airflow at the end of the swallow apnoea. These parameters were studied in eight healthy subjects under conditions of saliva swallowing and three doses of water (5,10 and 20 mL). The system allowed a real time description of the respiratory and feeding events, which were in close agreement with physiological principles. Mean results showed an increase of the apnoea time that was not statistically significant. In contrast, a highly significant increase of respiratory impedance during swallowing was observed (p < 0.0001). The described instrument does not use radiation. It shows itself particularly well suited for studies of deglutition physiology, including mechanisms involved in airway protection during swallowing. It can also be potentially useful contributing to easy clinical bedside evaluations and biofeedback procedures for the rehabilitation of paediatric and elderly patients. PMID- 19126973 TI - Computational hemodynamics in the human aorta: a computational fluid dynamics study of three cases with patient-specific geometries and inflow rates. AB - OBJECTIVE: To demonstrate the potential role of computational fluid dynamics (CFD) in therapeutic decision making for treatment of vascular pathologies of the human aorta. METHODS: CFD simulations with patient specific geometries and patient-specific inflow boundary conditions obtained with magnetic resonance imaging were performed in three cases: 1) mobile thrombus in the aortic arch in a patient with ischemic stroke 2) acute type II B aortic dissection 3) abdominal aortic aneurysm repaired with an endoluminal graft. Blood flow pathlines, wall shear stresses (WSS), dynamic pressures, blood velocities and flow particle resident times were calculated. RESULTS: Aortic thrombus was indicated as possible source of emboli by flowlines and elevated WSS (8% higher than average WSS at aortic wall) in case 1. This was not identified on conventional imaging. In case 2, the false lumen of the dissection showed elevated pressures and high blood velocities at systole but low pressures and stagnant flow at other times (blood velocity < 0.02 m/s and WSS < 0.1 Pa). Flow disturbances at the reentrance zone of blood from the false lumen were recognized. For case 3, elevated WSS at the landing zone of the endoluminal graft and at the right iliac section together with disturbed flow patterns and increased flow particle resident times were noted. Focal stenoses coincided with the flow disturbances. Measured velocity patterns were qualitatively in agreement with velocity patterns calculated with CFD. CONCLUSION: CFD simulations provide additional information of the hemodynamics in the diseased human aorta and may have potential in aiding the therapeutic decision making process. PMID- 19126974 TI - Response of a viscoelastic layer (mucus) to turbulent airflow in a rigid tube. AB - Basic interaction mechanism between the air flow and viscoelastic mucus layer lining a rigid tube is computationally studied. Linear wave instability theory is applied to the coupled air-mucus system to explore the stability of the interface. Primary velocity profile is taken to be the mean profile of turbulent flow and turbulent fluctuations are neglected. The model predicts that the instability initiates in the form of slow propagating waves on the mucus surface. Onset flow speed at which these waves initiate is very sensitive to mucus viscosity to elasticity ratio at lower range and it approaches to an asymptotic value for higher values. The results indicate that while the wave length increases, wave speed decreases with increasing mucus viscosity to elasticity ratio. Model also predicts that the waves initiate at lower flow velocities for the turbulent case compared to the published laminar case. Turbulent onset flow speed is only 34% Flow is considered to be turbulent during forced expiration and coughing in central and upper airways. Model predicts that this flow behavior tends to favor wave initiation at lower flow rates and may facilitate cough clearance. PMID- 19126975 TI - Mutual information-based approach to the analysis of dynamic electrocardiograms. AB - Dynamic electrocardiogram (ECG) plays an important role in the analysis of heart movement and cardio-diseases. In an attempt to obtain a better understanding of the mechanisms of patterns and differences found in dynamic ECGs, techniques based on different theories such as chaos and fractal theory have been used to extract nonlinear information encoding in dynamics ECG signals. In this paper, we propose an information theory approach to supporting the analysis of dynamic ECG recorded during different time of a day. Mutual information of R-R intervals extracted from four subject groups were calculated and analysed. Results indicate that heart movement is similar to chaotic movement in many ways. Moreover, the mutual information of R-R intervals exhibits different patterns over different periods of a day and different subject groups, suggesting that it would be a useful tool to support classification analysis of heart movement and cardio related diseases. PMID- 19126976 TI - Editorial: primers. PMID- 19126977 TI - Section 1: medical informatics and eHealth. PMID- 19126978 TI - Effect of difference of cupula and endolymph densities on the dynamics of semicircular canal. AB - The effect of different densities of a cupula and endolymph on the dynamics of the semicircular canals is considered within the framework of a simplified one dimensional mathematical model where the canal is approximated by a torus. If the densities are equal, the model is represented by Steinhausen's phenomenological equation. The difference of densities results in the complex dynamics of the cupulo-endolymphatic system, and leads to a dependence on the orientation of both the gravity vector relative to the canal plane and the axis of rotation, as well as on the distance between the axis of rotation and the center of the semicircular canal. Our analysis focused on two cases of canal stimulation: rotation with a constant velocity and a time-dependent (harmonically oscillating) angular velocity. Two types of spatial orientation of the axis of rotation, the axis of canal symmetry, and the vector of gravity were considered: i) the gravity vector and axis of rotation lie in the canal plane, and ii) the axis of rotation and gravity vector are normal to the canal plane. The difference of the cupula and endolymph densities reveals new features of cupula dynamics, for instance--a shift of the cupula to a new position of equilibrium that depends on the gravity vector and the parameters of head rotation, and the onset of cupula oscillations with multiple frequencies that results in the distortion of cupula dynamics relative to harmonic stimulation. Factors that might influence the density difference effects and the conditions under which these effects occur are discussed. PMID- 19126979 TI - Simulation of cupulolithiasis and canalolithiasis by an animal model. AB - The physical mechanisms responsible for cupulolithiasis and canalolithiasis have been investigated by two groups of experiments in isolated posterior semicircular canal (SCC) of frog (Rana esculenta L.). First, clouds of 10-30 isolated otoconia were let to fall (snowfall of otoconia) either through the ampulla onto the cupula, or inside the long arm of the canal, opposite to the cupula. Second, microspheres ranging 30 to 350 microm in diameter were gently moved to and fro inside the long arm of the canal by a micromanipulator. The effects were evaluated by recording the firing rate (Nfr) of the ampullary nerve. Snowfall of otoconia produced detectable changes of Nfr only when otoconia got in contact with the cupula, but not when falling through the endolymph. Movement of the microspheres in the canal long arm induced Nfr changes only if the microsphere diameter exceeded about 50 microm. Although the exact microsphere size needed for receptor stimulation may depend on the experimental conditions, these results strongly suggest that debris moving inside a SCC (canalolithiasis) can produce transcupular pressures able to stimulate ampullar receptors only if they have suitable size, whereas isolated otoconia cannot, except when lying on the cupula (cupulolithiasis). PMID- 19126980 TI - Orientation of Listing's plane after hypergravity in humans. AB - Adaptation to a novel gravitational state involves adaptation of vestibular mediated responses, in particular those mediated by the otolith organs. The present paper investigates whether the orientation of Listing's plane, which is under control of otolith signals, is affected by sustained exposure to hypergravity. Subjects were exposed to four G-loads differing in duration (45 or 90 min) and magnitude (2 or 3G). During centrifugation subjects were in a supine position, directing the gravito-inertial acceleration along the naso-occipetal axis. We determined the orientation of Listing's plane before and after each centrifuge run, with the head erect and tilted in pitch. Head tilt in pitch induced a counter-pitch of Listing's plane, which was found to be less pronounced after centrifugation. In addition, exposure to 3G for 90 min induced a small backward tilt of Listing's plane compared to the pretest orientation (head erect). In order to explain these results a hypothesis is discussed, proposing that the orientation of Listing's plane in the head is governed by a head fixed orientation vector that is modulated by the direction of gravity relative to the head. Sustained centrifugation is proposed to decrease this gravitational modulation, leading to the effects observed. This could reflect a shift towards a more body centered frame of reference. PMID- 19126981 TI - Time course of the change in subjective straight-ahead after vestibular stimulation. AB - We measured the change in subjective straight-ahead (SSA) as a function of time after vestibular stimulation. Blindfolded subjects were exposed to sudden stops after constant velocity (72 degrees/s) z-axis rotation. They were then asked to open their eyes and continuously adjust the position of a laser spot to the position perceived as straight ahead (visual localization of SSA) immediately, or after a 10 s or 20 s delay following the sudden stop. The deviation of SSA increased to 20 s in the opposite direction of acceleration and then decreased gradually when the visual localization task was started immediately after sudden stops. The time courses changed systematically when the task was started after a delay of 10 s or 20 s following sudden stops: the initial increase component and maximum magnitude of the SSA deviation fell off with delay length, but the increase-decrease pattern was preserved. These delay effects on the time course of the SSA deviation might reflect that the velocity-to-orientation/position integrator for perception of body orientation in vestibular system might be inactivated temporarily during the delay period after vestibular stimulation without visual input. PMID- 19126982 TI - Vestibulo-ocular (oVEMP) responses produced by bone-conducted sound stimuli applied to the mid-sagittal plane of the head. AB - Recently several studies have yielded evidence that impulses of bone-conducted (BC) sound can produce short-latency myogenic responses in the extraocular muscles, which are probably mediated by otolithic afferents. These responses, although miniscule, can be recorded with surface electrodes and are termed ocular vestibular evoked myogenic potentials (oVEMP). It is assumed that in response to low-frequency BC-sound stimuli the head moves predominantly along the axis from the site of the applied stimulus to the opposite side. Thus, oppositely-directed accelerations along a particular axis would produce oppositely-directed compensatory vestibulo-ocular responses (VOR) and oVEMPs. The aim of this study was to investigate whether the oVEMPs would reflect these direction-dependent VOR responses. Single cycles of 125 and 250 Hz BC tones were applied to opposite sides of two approximately orthogonal, naso-occipital (x) and vertical (z) axes of the head. oVEMP responses were recorded with standard bilateral vertical EOG montages. The responses in all twelve healthy subjects showed consistent differences with regard to the latency and/or shape of the response to stimuli applied to opposite sides of the head. These differences likely reflect different patterns of electro-myographic activity of the extraocular muscles, which may be mediated by groups of vestibular (probably otolithic) afferents with differently orientated spatial polarization vectors. PMID- 19126983 TI - Symptoms of detachment from the self or from the environment in patients with an acquired deficiency of the special senses. AB - To compare the frequency of symptoms of detachment from the self or from the environment (DD) in patients with an acquired deficiency of the special senses, a questionnaire for DD symptoms [12] and the GHQ12 questionnaire for common mental disorders [16] were auto-administered to patients with hearing-loss (N = 40), peripheral vestibular disease (N = 40) or bilateral retinal disease (N = 40), and to 80 healthy subjects. Patients with retinal disease and patients with vestibular disease reported DD symptoms more frequently & severe than patients with hearing loss and healthy subjects. DD scores were related to the GHQ12 scores and to the type of sensory dysfunction. DD symptoms are more frequent & severe in patients with an acquired deficiency of the special senses, associated to symptoms of common mental disorders. PMID- 19126984 TI - Aquatic physiotherapy for vestibular rehabilitation in patients with unilateral vestibular hypofunction: exploratory prospective study. AB - PURPOSE: To explore the effects of aquatic physiotherapy on individuals with unilateral vestibular hypofunction and examine the association of therapeutic effects with age, time since symptom onset and use of antivertigo medication. STUDY DESIGN: Exploratory study. SETTING: Tertiary referral center. PATIENTS: Twenty-one patients presenting chronic dizziness with uncompensated unilateral vestibular loss. INTERVENTION: All patients underwent a regimen of 10 sessions of aquatic physiotherapy for vestibular rehabilitation. MAIN OUTCOME MEASURES: Patient evaluation prior to and following rehabilitation involved the application of the Brazilian version of the Dizziness Handicap Inventory (DHI), dynamic computerized posturography and the self-perception scale of dizziness intensity. Statistical analysis compared average variables prior to and following rehabilitation using the Student t test. RESULTS: Brazilian DHI total scores were lower following rehabilitation (p = 0.001). Posturography revealed a reduction in the variation of body displacement following rehabilitation in the average stability and anterior/posterior stability indexes (p = 0.001) and in the average medial/lateral stability index (p = 0.003). Self-perception of dizziness intensity was lower following rehabilitation (p = 0.001). No association between age, time since symptom onset and use of antivertigo medication and rehabilitation therapeutic effects were found. CONCLUSION: Unilateral vestibular hypofunction patients undergoing aquatic physiotherapy for vestibular rehabilitation achieved an improvement in quality of life, body balance and self perception of dizziness intensity, regardless of age, time since symptom onset, and use of antivertigo medication. PMID- 19126986 TI - The reliability and validity of the Vertigo Symptom Scale and the Vertigo Dizziness Imbalance Questionnaires in a Turkish patient population with benign paroxysmal positional vertigo. AB - The aim of this study was to adapt the Vertigo Symptom Scale (VSS) and the Vertigo Dizziness Imbalance (VDI) Questionnaires to the Turkish population and investigate the reliability and validity of the Turkish version. One hundred and three patients with Benign Paroxysmal Positional Vertigo (BPPV) were included in the study. The patients were evaluated with the VSS and the VDI twice, at recruitment and 24 hours later. To perform concurrent validity study study, patients were also assessed by the Beck Depression Inventory (BDI) and the Medical Outcomes Survey Short Form 36 (SF-36) which were formerly proved to be valid and reliable for use in Turkish population. The internal consistency of the VSS, VDI-symptom scale, and VDI-health-related quality of life scale were good with Cronbach's alpha of 0.91, 0.85, and 0.93 respectively which showed high reliability for the Turkish versions. Test-retest reliability was also good, with a high intraclass correlation coefficient (ICC) between the two time points; being 0.83 for the VSS, 0.90 for the VDI-symptom scale and 0.89 for the VDI health related quality of life scale. Regarding concurrent validity, significant expected correlations were detected between the VSS and BDI (r = 0.55 p < 0.001) and the VSS and SF-36 (r = -0.43 p < 0.001). Significant low correlations were detected between the VDI-SS and BDI (r = 0.20 p < 0.05) and the VDI-SS and SF-36 (r = -0.21 p < 0.05). High correlations were observed between the VDI-HRQoL and BDI (r = -0.75 p < 0.001) and the VDI-HRQoL and SF-36 (r = 0.82, p < 0.001) which indicates high concurrent validity. The correlation between VDI-SS subscale scores and VDI-HRQoL subscale scores were 0.028 (P = 0.778) at the first time point. From the perspective of discriminant validity, it means that the VDI-SS measures a construct different from the one underlying the VDI-HRQoL. The adaptation of the VSS and VDI to the Turkish population was successful and both scales were found to be valid and reliable. Thus, they can be used in Turkish people with BPPV for assessment and monitoring the treatments. Besides, the results of the national studies in which these Turkish versions are used can be compared with those of the international studies. PMID- 19126985 TI - Effect of vestibular rehabilitation on passive dynamic visual acuity. AB - While active dynamic visual acuity (DVA) has been shown to improve with gaze stabilization exercises, we sought to determine whether DVA during passive head impulses (pDVA) would also improve following a rehabilitation course of vestibular physical therapy (VPT) in patients with unilateral and bilateral vestibular hypofunction. VPT consisted of gaze and gait stabilization exercises done as a home exercise program. Scleral search coil was used to characterize the angular vestibulo-ocular reflex (aVOR) during pDVA before and after VPT. Mean duration of VPT was 66 +/- 24 days, over a total of 5 +/- 1.4 outpatient visits. Two of three subjects showed improvements in pDVA with a mean reduction of 43% (LogMAR 0.58 to 0.398 and 0.92 to 0.40). Our data suggest improvements in pDVA may be due in part to improvements in aVOR velocity and acceleration gains or reduced latency of the aVOR. Each subject demonstrated a reduction in the ratio of compensatory saccades to head impulses after VPT. Preliminary data suggest that active gaze stability exercises may contribute to improvements in pDVA in some individuals. PMID- 19126987 TI - Transcutaneous pO2 measurement during tourniquet-induced venous occlusion using dynamic phosphorescence imaging. AB - A sufficient oxygen supply in skin grafts requires a functioning microcirculation. Venous occlusion impairs the microcirculation and is therefore a major threat of healing. Luminescence life time imaging (LLI) enables the non invasive and two-dimensional assessment of the transcutaneous oxygen partial pressure (p(tc)O2). In the current trial this new device was applied for monitoring of venous congestion. A tourniquet on the upper arm was inflated up to 40-50 mmHg and released after 10 min in eight healthy volunteers. The p(tc)O2 was measured at the lower arm every minute prior to, during and up to 10 min after cuff occlusion (40 degrees C applied skin temperature) using LLI of platinum(II) octaethyl-porphyrin immobilized in a polystyrene matrix. For validation the polarographic Clark electrode technique was applied in close proximity and measurement was performed simultaneously. p(tc)O2 measurements prior to (Clark: 50.68+/-5.69 mmHg vs. LLI: 50.89+/-4.96 mmHg) and at the end of the venous congestion (Clark: 16.41+/-4.54 mmHg vs. LLI: 23.82+/-3.23 mmHg) did not differ significantly using the Clark electrode vs. LLI. At the initial congestion respectively reperfusion phase the Clark electrode measured faster decreases respectively increase of p(tc)O2 due to oxygen consumption of this method. This experimental trial demonstrates the applicability of LLI to quantify the p(tc)O2 under changing venous blood flow. The use of planar transparent sensors allows the non-invasive generation of two-dimensional maps of surface pO2 what makes this method particular suitable for monitoring of skin grafts. PMID- 19126988 TI - Effect of disodium monofluorphosphate on plasma and blood viscosity in the rat. AB - Alpha macroglobulins (AM) are plasma proteins whose main function is to inactivate proteinases, protecting the tissues from the action of these enzymes. AM have influence on plasma viscosity (PV) and binds monofluorophosphate (MFP), which disturbs its homeostasis. The aim of this work was to evaluate whether the administration with MFP could modify blood viscosity. AM levels (micromol/l), PV (mPa.s), viscosity of red blood cells suspensions in NaCl 9 g/l (VES) and in autologue plasma (VEP) were measured in fifty-day old rats after a single dose of 80 micromol MFP or after 30 days of treatment with 80 micromol of MFP. Relative viscosity (RV) was calculated as the ratio VEP/PV. AM and PV increased significantly after 30 min of an oral dose of MFP. Controls (n=6), AM: 19.65+/ 0.85, PV: 1.39+/-0.01, treated (n=6), AM: 22.88+/-0.75 (p<0.05), PV: 1.76+/-0.14 (p<0.05). After 30 days of treatment with MFP, AM and PV increased significantly. Controls (n=6), AM: 10.76+/-1.33, PV: 1.19+/-0.04, treated (n=6), AM: 17.66+/ 1.27 (p<0.05), PV: 1.38+/-0.03 (p<0.05). The treatment with MFP modifies neither the VEP nor the RV. These results would indicate that AM and/or MFP did not interact with erythrocyte membrane and did not modify erythrocyte deformability. PMID- 19126989 TI - Imaging of aortic lesions with color coded duplex sonography and contrast enhanced ultrasound versus multislice computed tomography (MS-CT) angiography. AB - PURPOSE: The purpose of this study was to compare color coded duplex sonography (CCDS), contrast-enhanced ultrasound (CEUS) and multislice computed tomography (MS-CT) angiography in pathological aortic lesions. MATERIAL AND METHODS: 36 patients with a mean age of 71 years (range 51-87 yrs) with known or suspected treated and untreated aortic lesions detected by CTA were included in this prospective study. Standardized MS-CTA using a 16 or 64 row scanner (Somatom Sensation 16 or 64, Siemens Medical Systems, Forchheim, Germany) served as reference standard. The patients were examined with CCDS and CEUS (Logiq 9, General Electrics, Milwaukee, WI) using the low MI technique with intravenous injection of 1.6-2.4 ml SonoVue (Bracco, Italy). Contrast enhanced ultrasound was performed in the arterial, venous and late venous phase. RESULTS: In all patients CEUS resulted in a relevant diagnosis of an aortic pathology. In total, two covered ruptures, nine aortic dissections and five aortic aneurysms were detected. In the routine follow-up of 20 patients following endovascular repair (EVAR) of abdominal aortic aneurysm (AAA), one type 3 and three type 2 endoleaks after y-stent graft were detected. For presurgical planning, in addition to the perfused true and false lumen after dissection the perfusion of the renal arteries was examined. CONCLUSION: CEUS with SonoVue allows the evaluation of aortic lesions. It is an additional examination to CTA. It may allow a more rapid and noninvasive diagnosis, especially in critical patients from intensive care units because of its bedside availability. As the examination is dynamic, additional information about blood flow in the true and false lumen and about renal perfusion after dissections can be evaluated. Especially in patients with contraindications to CT contrast agents (e.g. due to renal failure or severe allergy), CEUS provides a good alternative to MS-CT. PMID- 19126990 TI - Effects of cardiotonic pill on RBC rheologic abnormalities in HFD-induced mice and LPL deficient mice. AB - The lipoprotein lipase deficient (LPL-/-) mice and high fat-diet (HFD) induced hypertriglyceridemic mice were used to investigate the effects of cardiotonic pill (CP) on RBC rheologic abnormalities. Mice were randomly divided into the following five groups: the control group; the untreated HFD group; the untreated LPL-/- group; the treated HFD group; and the treated LPL-/- group, and the treated HFD and LPL-/- mice were administered with CP twice a day (400 mg/kg/day) orally for four weeks. Then, plasma triglyceride (TG), RBC deformation index (DI), orientation index (DI)or and RBC electrophoretic time (EPT) were measured. Compared with the untreated HFD mice, TG level and EPT reduced and DI and (DI)or increased markedly in the treated HFD mice (P<0.05). However, compared with the untreated LPL-/- mice, these parameters in the treated LPL-/- mice had no statistically significant changes (P>0.05). Our data show that CP can lower plasma TG level and ameliorate RBC rheologic abnormalities in the HFD-induced hypertriglyceridemic mice, but it losses its capacity in the LPL deficient animals. The results indicate that LPL may be one of the important targets for CP regulating lipometabolism and rheologic abnormalities. PMID- 19126991 TI - Erythrocyte membrane composition in patients with primary hypercholesterolemia. AB - There are conflicting results regarding the erythrocyte membrane cholesterol and phospholipid content in patients with primary hypercholesterolemia (PHC), due to methodological problems in obtaining haemoglobin-free ghosts. At the same time, the different units used and the fact that the cholesterol and phospholipids are not expressed in relation with integral protein membrane content, produces contradictory results. We have analysed in 33 patients with PHC (12 male, 31 female) aged 43+/-12 years and in 33 healthy normolipaemic volunteers (9 male, 24 female) aged 43+/-13 years plasma lipids, along with, erythrocyte membrane cholesterol, phospholipids and integral proteins. PHC patients showed increased erythrocyte membrane cholesterol: 0.36+/-0.15 mg/mg when compared with controls: 0.29+/-0.75 mg/mg; p=0.018. Phospholipid membrane content, although higher in the cases, did not reach statistical significance (PHC patients: 0.38+/-0.15 mg/mg vs. 0.33+/-0.72 mg/mg; p=0.098). The cholesterol/phospholipids ratio (Chol/Ph) was 0.99+/-0.22 in PHC patients versus 0.92+/-0.28 in controls; p=0.127. Our results suggest that there is a slight increase in erythrocyte membrane cholesterol in patients with PHC. Given the increasing importance of erythrocyte membrane cholesterol in the stability of the atheroma plaque due its possible contribution to the clinical signs of ischaemic heart disease, it seems relevant to determine this parameter in risk populations. Therefore, a simple and reproducible method needs to be standardised which would enable comparisons between laboratories and facilitate further studies aimed to it as a marker of acute coronary syndromes. PMID- 19126992 TI - Relation of platelet aggregation and fibrinogen levels to advancing age in aspirin- and thienopyridine-treated patients. AB - In our present study we investigated the association between platelet aggregation in patients treated with the most widely used antiplatelet agents (100 and 300 325 mg acetylsalicylic acid (ASA), 75 mg clopidogrel, 500 mg ticlopidine and the combination of 100 mg aspirin and 75 mg clopidogrel), fibrinogen levels and aging. Between 2001 and 2005 we measured in vitro platelet aggregation in 5026 vascular patients according to the method of Born. Platelet aggregation was tested with 5 and 10 microM adenosine-diphosphate, 2 microg/ml collagen and 10 microM epinephrine stimulants. Fibrinogen level was simultaneously measured in a subgroup of 3243 patients. The subjects were divided by age into decades. Platelet aggregation increased significantly with advancing age in the case of 100 and 300-325 mg ASA-treated patients (p<0.001). In aspirin-treated patients also fibrinogen levels increased with aging (p<0.001). There was no association between platelet aggregation or fibrinogen levels and aging either in patients treated with 75 mg clopidogrel or with 500 mg ticlopidine. Thienopyridine-treated patients exhibited significantly lower fibrinogen levels than ASA-treated individuals (p<0.001). Our results suggest that advancing age is associated with elevated platelet aggregability in widely used antiplatelet regimens that might contribute to higher risk of cardiovascular events in the elderly. PMID- 19126993 TI - Oxygen delivery index in subjects with normal haemoglobin (HbAA), sickle cell trait (HbAS) and homozygous sickle cell disease (HbSS). AB - Sickle cell disease is characterized by altered blood rheology due to a reduced haematocrit and a resulting lowered viscosity. Oxygen carriage, and consequently oxygen delivery, may be deleteriously affected if the haematocrit reduction is such as to limit oxygen uptake from the lungs and delivery to the tissues. The present study seeks to determine and compare the oxygen delivery index (ODI) in subjects with normal and abnormal haemoglobin genotypes. Thirty four apparently healthy subjects having normal haemoglobin genotype (AA), 27 with sickle cell trait (AS) and 50 with homozygous sickle cell disease (SS) were recruited into the study. Whole blood viscosity was measured at low and high shear rates of 23 s(-1) and 230 s(-1), respectively, using a Wells Brookfield Cone and Plate Viscometer. Haematocrit was determined using an AC.Tron Coulter Counter. The oxygen delivery index was calculated as the ratio of the haematocrit to whole blood viscosity. There was a statistically significant difference in the ODI in the SS group compared with both the AA and AS groups. There was no statistical significance in the ODI between the AA and AS groups. The ODI may be considered as a useful assessment of oxygen delivery in subjects with sickle cell disease. PMID- 19126994 TI - Evaluation of the microcirculation during extracorporeal membrane-oxygenation. AB - Extracorporeal membrane oxygenation (ECMO) provides pulmonary and circulatory support in critically ill patients (Cardiopulmonary Resuscitation, Acute Respiratory Distress Syndrome, or low output syndrome). Hemodynamic parameters are used for evaluation of the macrocirculation, while the microcirculation is monitored by blood-lactate as a surrogate parameter. We evaluated the microcirculation by orthogonal polarization spectral imaging in a patient during ECMO support. This method was initially proposed to quantify changes of microcirculation in patients with septic shock. However, we were able to non invasively monitor microcirculatory changes at the bedside during temporary intentional arrest of ECMO due to an exchange of the oxygenator. Using a computerized analyzation model, the flow after ECMO stop in vessels (10-100 microm) in the sublingual mucosa was acutely absent or intermittent, respectively. 120 s after restart, microflow was improved with new ECMO settings compared to baseline, while macrocirculation with a mean arterial pressure of 75 mmHg was present after 60 s. The application of orthogonal polarization spectral imaging might be a valuable technique for evaluation of the microcirculation during extracorporeal circulation. It is rapidly implementable, can be used in vivo, and no invasive probes are required. PMID- 19126995 TI - Susceptibility of oxidative stress on red blood cells exposed to gamma rays: hemorheological evaluation. AB - Irradiation has been shown to induce biochemical changes in stored red blood cells (RBCs) and to generate reactive oxygen species (ROS). This study evaluated the hemorheological properties, the degree of lipid peroxidation and the oxidative susceptibility of irradiated RBCs. Furthermore, we investigated the radioprotective role of N-t-butyl hydroxylamine (NtBHA) against gamma-ray exposure of RBCs. RBC concentrates were irradiated with a minimum dose of 25 Gy, and were exposed to FeSO4 to examine the oxidative susceptibility. RBC deformability was evaluated by the use of a microfluidic ektacytometer, in relation to the hematological and biochemical properties. The deformability of the irradiated RBCs was significantly lower than that of control. Exposure to gamma rays significantly increased the mean corpuscular volume (MCV) and lipid peroxidation. Changes in RBC deformability were more prominent in irradiated RBCs than in non-irradiated RBCs also under conditions of oxidative stress. The deformability of NtBHA treated RBCs prior to irradiation was not altered as compared with irradiated RBCs not treated with NtBHA. In conclusion, irradiation reduces RBC deformability during storage and the irradiated RBCs seem susceptible to oxidative stress. NtBHA may not have a protective role against the effects of gamma-ray exposure in RBCs but further evaluation of NtBHA or another radioprotective compound is required. PMID- 19126996 TI - Short-term effects of atorvastatin on hemorheologic parameters and endothelial dysfunction in patients with hypercholesterolemia. Lower is better? PMID- 19126998 TI - Assessment, treatment and service issues for students with traumatic brain injury. PMID- 19126999 TI - A holistic brain injury rehabilitation program for school-age children. AB - In the field of pediatric neurorehabilitation, there is a dearth of well-defined models of care for children with brain injuries and research to support them. Literature reviews of both adult and pediatric studies suggest that programs involving a comprehensive and holistic approach to care may be particularly effective in addressing the issues faced by individuals recovering from acquired brain injury. The primary purpose of this paper is to propose a model of holistic neurorehabilitation for children with an acquired brain injury, and to describe its theoretical bases. The paper describes such a program developed at Children's Specialized Hospital in New Jersey, USA, and presents a brief synopsis of the program's philosophy, components, goals, and therapeutic interventions. In addition, the paper discusses the type of data necessary to demonstrate the clinical utility and cost-effectiveness of such a program, and calls for future research to examine this. PMID- 19127000 TI - Self-regulation abilities in children with severe traumatic brain injury: a preliminary investigation of naturalistic action. AB - Research suggests that the occurrence of a traumatic brain injury (TBI) in childhood may disrupt self-regulation abilities, putting children at risk for difficulty on everyday tasks requiring self-regulation throughout their development. In the current exploratory study, a novel age-appropriate task assessed the ability to perform three familiar tasks using real objects while adhering to specific rules. Performance of children (ages 8-16) with severe TBI (n = 11) on the naturalistic task was compared to that of typically developing children (n = 21), including measures of the amount/types of errors and number of broken rules. The children with TBI exhibited significantly increased use of distractor objects in place of target objects as compared to the non-injured children. Additionally, children with TBI demonstrated trends of increased breaking of rules during the task and failure to include necessary steps. The preliminary results support the theory that children with severe TBI possess inefficient supervisory processes of self-regulation, corresponding to a decreased ability to carry out goal-based top-down processing. They may instead exhibit a bias towards a bottom-up approach, depending primarily on environmental cues such as the objects present to guide their actions, thus impeding self regulation abilities. PMID- 19127001 TI - Return to school following childhood TBI: who gets services? AB - Long-term follow-up studies of children conducted during the K-12 school years suggest that problems associated with TBI tend to persist or worsen as children progress through school. A critical issue in service delivery for students with TBI is the significant discrepancy between the incidence of TBI and the identification of children with TBI for special education services. The purpose of this study was to examine factors that influence identification and service practices for students with TBI. Parents of 56 youth with TBI completed a questionnaire and interview about the hospital-school transition and educational services provided within the first year of school re-entry. A subsample participated in a concurrent qualitative study. Results document that 25% of the sample were identified for formal services (via individual education plan or 504 plan) while over 41% received informal supports (e.g., schedule change, extra time on tests). Results from chi-square and logistic regression analyses suggest that injury severity and hospital-school transition services (e.g., written or verbal communication between hospital and school) were related to the provision of formal special education or 504 services. A critical factor contributing to the identification of students with TBI for special education is the link between hospital and school. PMID- 19127002 TI - Practical applications for use of PDAs and smartphones with children and adolescents who have traumatic brain injury. AB - It is often recommended that individuals with memory and organizational problems after a traumatic brain injury (TBI) use some type of memory device such as a paper calendar. Recently the use of electronic devices has been suggested. This article outlines data obtained from in vivo trials using personal data assistants (PDAs) and follow up in depth studies with PDAs and smartphones. These trials were conducted with individuals who had memory and organizational problems as a result of cognitive disorders of traumatic brain injury (TBI) or intellectual disability (ID) Results indicate that the use of electronic devices can enhance independent behavior. Factors influencing success include: student motivation, audible beep of the device; support for programming and troubleshooting, alterations of functions; and selection of features to motivate. Based on the result of the studies, an intervention plan for use of PDAs was developed for use by clinicians. PMID- 19127003 TI - Measuring social cognition in adolescents: implications for students with TBI returning to school. AB - In everyday adolescent communication, the ability to empathise with the mental state of others, recognise or infer intentions, or make judgements about emotional state, is a non-conscious but vital prerequisite of relating. Execution of these skills in social interactions supports both the exchange of social knowledge and also the development and maintenance of personal relationships. Thus, adolescents with impairments in these skills are at risk for a variety of negative outcomes. In this paper, we present data to illustrate that adolescents with traumatic brain injury (TBI) are likely to have impairments in processes such as emotion recognition and mental state attribution, and that these might not be identified on standardised tests. This is considered from the perspective of clinical assessment and intervention in school contexts. PMID- 19127004 TI - An electronic survey about college experiences after traumatic brain injury. AB - For many with traumatic brain injury (TBI), going to college is a realistic goal; however there is little documentation of the challenges faced by those with TBI who attend college. The primary purposes of this study were to document the academic challenges (studying, in-class experiences, time management, psychosocial aspects) reported by adults with TBI, and to investigate relationships between these challenges and the physical, cognitive and psychosocial consequences of TBI. An electronic anonymous survey was distributed. Of the 35 respondents with TBI, nearly all reported the need to review material more and a majority reported that others do not understand their problems. In class experiences of being nervous before tests, forgetting what is said in class, and getting overwhelmed in class were also reported by a majority. Those who reported more physical, cognitive and psychosocial consequences of their injury also identified more academic challenges as well, although cognitive consequences alone predicted academic challenges better than all of the consequences combined. Psychosocial aspects also predicted academic challenges to a lesser extent. In spite of these findings, nearly half of the respondents had not heard of or had never accessed campus disability services. To develop effective on-campus service delivery models, additional research is needed to understand why students do or do not make use of existing services. PMID- 19127005 TI - Assessing readiness for post-secondary education after traumatic brain injury using a simulated college experience. AB - Approximately 1.4 million Americans sustain a traumatic brain injury (TBI) each year with approximately two-thirds of these injuries affecting people between the ages of 15 and 24 years. Consequently, many adults with brain injury sustain their injuries during a time when they are making important decisions regarding future vocational activities, including return to school. Cognitive impairments resulting from TBI present as a significant barrier to return to school and it is difficult to predict those persons who will succeed in school after a TBI and those who will not. This article presents a college simulation experience currently used in a rehabilitation setting. It involves lectures and examinations that test an individual's ability to learn new material. Lecture content focuses on the nature and effects of brain injury and on study skills. In addition, compensatory strategies commonly used in university settings (note takers, extended time for exams, taped lectures) may be used to assess potential benefit of these services. The simulated college experience provides a realistic appraisal of the individual's ability to learn information in a lecture-oriented class setting. It also provides information regarding the benefits of specific study skills and compensatory strategies. Case studies of three individuals with severe TBI are presented. PMID- 19127006 TI - Performance of three racial/ethnic groups on two tests of executive function: clinical implications for traumatic brain injury (TBI). AB - The purpose of this study was to determine if race/ethnicity may influence performance of college students on the Behavioral Assessment of Dysexecutive Syndrome (BADS) and the Wisconsin Card Sorting Test (WCST), two different tests of executive function (EF). A total of 149 participants who self-identified as African American (AA), European American (EA), and Latina/o American (LA) constituted the three different racial/ethnic groups. Participants ranged in age range from 18 to 24 years old (SD = 1.29) and none reported histories of disabilities. The BADS was administered to 149 college students and the WCST was administered to 142 of the same participants. No statistically significant differences were found for race/ethnicity or gender on the WCST. Statistically significant differences were found for race/ethnicity on the Profile Scores (raw scores), standard scores (SS) and the Zoo Map subtest of the BADS, but no statistically significant differences were found for gender on these same tasks. While there should be less concern in administering WCST to diverse groups who are English speakers, caution should be taken in administering and interpreting the BADS for individuals from diverse cultural and linguistic backgrounds. PMID- 19127007 TI - Comparison of ergonomic risk assessment output in a repetitive sawmill occupation: trim-saw operator. AB - Multiple ergonomic risk assessment methods of unique structure are currently being used to direct industrial prevention initiatives focused on musculoskeletal injuries. In this study, the physical exposures required to perform an at-risk sawmill occupation were collected from 29 subjects via quantified means (surface electromyography and electrogoniometery) and used to calculate several ergonomic risk assessment methods. The aims of this study are to: 1) compare the output of the RULA, REBA, ACGIH TLV, Strain Index and OCRA ergonomic risk assessment methods, 2) examine the assessments' ability to differentiate between facilities reporting meaningfully different incidence rates, and 3) examine the effect of varying the definition of end range posture and exertion required on risk assessment scores. Risk level output assigned by all methods were not sensitive to inter facility differences in risk of injury, suggesting interpretation of risk index and component scores are needed to direct intervention. Components of all methodologies were sensitive to worker technique and facility assessed. Varying variable definition resulted in significantly different component, combined component and/or risk output scores in all methods assessed. The significant effect of posture and exertion variable definition suggests definitions taken to be interchangeable by work site evaluators are not equivalent. PMID- 19127008 TI - Age, work limitations and physical functioning among construction roofers. AB - BACKGROUND: To investigate the intersection of aging with work limitations, chronic medical and musculoskeletal conditions, and physical functioning we undertook a cross-sectional study of U.S. construction roofers who were current union members between the ages of 40 and 59. METHODS: Participants were asked about the presence of medical conditions and musculoskeletal disorders (MSDs); the Work Limitations Questionnaire, the SF-12, and other validated assessments of social and economic impact of injury were included. RESULTS: Sixty-nine percent had at least one of these conditions in the previous two years; 31% missed work. Workers with medical and musculoskeletal conditions were older, had the highest prevalence of work activity limitations, and had the lowest SF-12 scores. CONCLUSIONS: Older age was associated with the presence of a medical condition, and with reduced physical functioning. Medical and musculoskeletal conditions were strongly associated with work limitation, missed work, and reduced physical functioning. Older workers may be at higher risk of disability retirement compared to younger workers with similar medical conditions and work limitations. PMID- 19127009 TI - Occupational driving as a risk factor in low back pain: a case-control study in a Mexican population. AB - The objective of this study was to assess and quantify the degree to which interaction between occupational driving and lifting tasks is a risk factor in lumbar spondyloarthrosis etiology. A case-control study was performed with 231 workers, 18-55 years old, insured by the Mexican Social Security Institute (IMSS, according to its designation in Spanish). A multivariate analysis using conditional logistical regression showed that driving tasks, when combined with lifting tasks, are associated with this illness (OR = 7.3; 95% CI 1.7-31.4). Occupational driving as it interacted with daily lifting frequency resulted in a greater risk (OR = 10.4; 95% CI 2.0-52.5). No exposure-response relationship was found with daily hours spent working as a driver. The attributable risk for driving tasks was 0.86, suggesting that 86% of lumbar spondyloarthrosis could be decreased if risk factors were reduced through ergonomic redesign of the workplace and Manual Materials Handling (MMH) tasks, along with development of educational programs. PMID- 19127010 TI - Physical symptoms and psychological health status by the type of job. AB - BACKGROUND: The physical symptoms and mental health status of 3,447 men working in a dairy product company were evaluated by a health questionnaire, the Todai Health Index (THI). The purpose of this study is to determine the relationship between job content and subjective health status. METHOD: The authors conducted a cross-sectional study. Thirty-three branches of the company are distributed throughout Japan. The subjects ranged in age from 18 to 59 years old. The THI is composed of 130 questions and each subject's physical and mental health status was evaluated based on the scores in 13 specific scales. The subjects' jobs were classified into three categories: 1 production process work; 2 clerical or technical work; 3 transportation or sales work. Subjects grouped into the clerical or technical work category were set as the control group, because the authors speculated that the lowest workload was observed in group of clerical or technical work. RESULTS: The scores in each of the THI scales were compared among workers in the three job categories stratified by the age group (10-year age groups). The transportation or sales workers began to show high scores for irregularity of lifestyle from their 30's, and this was maintained until subjects' retirement from the company. In addition, the same group also began to show higher scores for physical symptoms, such as vague complaints, respiratory complaints, and eye complaints from their 40's and higher scores for mouth and evacuation and digestive complaints from their 50's. CONCLUSIONS: Transportation or sales workers should receive adequate health care to prevent a high prevalence of physical and/or illness. PMID- 19127011 TI - Development of risk assessment tool for foundry workers. AB - Occupational ill-health and work-related disorders are predominant in manufacturing industries due to the inevitable presence of manual work even after several waves of industrial automation and technological advancements. Ergonomic risk factors and musculoskeletal disorders like low-back symptoms have been noted amongst foundry workers. OBJECTIVE: The purpose of this study was to formulate and develop a Physical Effort Index to assess risk factor. SCOPE: The questionnaire tool applicable to foundry environment has been designed and validated. The data recorded through survey across the foundries has been subjected to regression analysis to correlate between proposed physical effort index and the standard Borg's Ratings of Perceived Exertion (RPE) scale. RESULTS: The physical efforts of sixty seven workers in various foundry shop floors were assessed subjectively. The 'Job factors' and 'Work environment' were the two major parameters considered in assessing the worker discomfort level at workplace. A relation between Borg's RPE scale and the above two parameters were arrived at, through regression analysis. CONCLUSIONS: The study demonstrates the prevalence of risk factors amongst foundry workers and the effectiveness of the proposed index in estimating the risk factor levels. RELEVANCE TO THE INDUSTRY: The proposed tool will assist foundry supervisors and managers to assess the risk factors and helps in better understanding of the workplace to avoid work-related disorders, ensuring better output. PMID- 19127012 TI - Perception of shoulder muscular effort during low-demand load transfer tasks. AB - This study focused on quantifying the mathematical relationship between shoulder physical loading and muscular effort perception during low physical demand tasks. Subjects underwent training to calibrate to their range of shoulder strength capability. Subjects transferred visually identical bottles representing specified percentages of extended arm maximal voluntary force (MVF) in defined azimuth directions to identified targets. They then reported their percentage of perceived shoulder exertion relative to their calibrated range. Measures of physical shoulder loading were calculated from experimental data with a dynamic shoulder moment model. Shoulder reported perceived muscular exertion (RPE) values were most significantly correlated with percent MVF (r = 0.81), suggesting subjects were influenced more by the manipulated hand load than the shoulder specific physical load. Multiple regression analyses demonstrated that other personal and task factors influenced shoulder RPE. Generally, subjects overestimated shoulder physical loading, and the quality of their perception degraded as the load increased. PMID- 19127013 TI - Job demands and health complaints in white and blue collar workers. AB - BACKGROUND: General health in the working population is thought to depend on working conditions. OBJECTIVE: This survey studied job demands and health complaints in working white and blue collar employees. We expect physical and psychological job demands to be differentially distributed among white and blue collar workers. Do they report health complaints consistent with their working conditions? METHOD: Cross-sectional study of 323 white and 383 blue collar workers. They completed the Basic Occupational Health Questionnaire, a valid and reliable self-report questionnaire about health, work and working conditions. The results were analysed using Chi-square and logistic regression analysis, controlling for educational level as a proxy of socioeconomic status. RESULTS: The questionnaires of 280 white and 251 blue collar workers were suitable for analysis. White collar workers reported higher psychological job demands, and blue collar workers reported higher physical demands. In both occupational groups, low back pain, fatigue and upper respiratory complaints were most common. The rates of low back pain and pain in the lower extremity were higher in blue collar workers, as were regular headaches, pain in the cardiac region and feeling sleepy. However, these relationships substantially weakened when the educational level was adjusted for. CONCLUSIONS: Despite the differential distribution of job demands, white and blue collar workers reported similar health complaints. Health in the working population depended predominantly on socioeconomic status. Interventions to improve general health of employees should be directed at their socioeconomic position instead of working conditions. PMID- 19127014 TI - Predictors of employment for young adults with developmental motor disabilities. AB - OBJECTIVE: To identify the personal, family, and community factors that facilitate or hinder employment for young adults with developmental motor disabilities. METHODS: Quantitative methods with an embedded qualitative component were used. Seventy-six persons between the ages of 20 and 30 years of age (Mean = 25, SD = 3.1) with a diagnosis of either cerebral palsy or spina bifida completed questionnaires addressing factors such as depression, and participated in a semi-structured interview that allowed participants to describe their experiences with education, employment, transportation, and other services. RESULTS: Almost half of the participants (n = 35) were not currently employed. Hierarchical regression analyses indicated that gender (females were less likely to be employed), IQ (lower IQ associated with unemployment), and transportation dependence accounted for 42% of the variance in employment. Themes emerging from content analysis of the interviews supported the findings related to transportation barriers. Social reactions to disability limited employment opportunities, and participants often felt stuck in terms of employment options with limited opportunities for advancement. CONCLUSIONS: Transportation is a significant barrier to employment and innovative solutions are needed. Issues related to gender need to be considered when addressing employment inequities for persons with primarily motor disabilities. PMID- 19127015 TI - Organisational change, job strain and increased risk of stroke? A pilot study. AB - AIMS: The objective of this pilot study was to explore whether organisational change and work-related stress, as measured by the Job Content Questionnaire, were associated with first-ever stroke among working people aged 30-65. METHODS: In a case-control study a total of 65 consecutive cases, aged 30-65 years of age, with first-ever stroke were recruited from four hospitals in Sweden during 2000 2002. During the same period, 103 random population controls in the same age interval were recruited. Data on job-related stress and traditional medical risk factors were collected by a questionnaire. RESULTS: In the multivariate analyses, organisational change (OR 3.38) increased the likelihood of stroke, while experiencing an active job (OR 0.37) decreased the likelihood of stroke. Regarding risk factors outside work, age (OR 1.11), low physical activity (OR 5.21), low education (OR 2.48) and family history of stroke (OR 2.59) were associated with increased likelihood of stroke. CONCLUSION: This study suggests an association between organisational change, work-related stress and stroke. The likelihood of stroke was lower for people in active job situations. PMID- 19127016 TI - Swedish social insurance officers' experience of their working conditions when dealing with applications for disability pension. AB - The aim of this study was to describe how Swedish social insurance officers experienced their working conditions, when dealing with applications for disability pensions (DPs). A qualitative research design was selected to describe their experiences. Open-ended interviews with ten social insurance officers were analyzed with inductive content analysis. This analysis showed that the working conditions were influenced by rules and regulations and the social insurance officers' competence. The social insurance officers' powerful positions and how their discretion was implemented made them feel responsible for performing their work well. They felt that the large number of clients hampered them from being able to offer the clients the activities they were entitled to, and from paying each client sufficient attention. There was a need for time to reflect and to improve management, in order to better develop the way in which DPs were handled. There was a difference in culture between the older social insurance officers, who had experience-based training, and the younger social insurance officers, who had academic qualifications. The results from this study could be used to better develop the organisation of work in the social insurance agency, and in further educational activities. PMID- 19127017 TI - The effect of body mechanics education on the work performance of fruit warehouse workers. AB - Agriculture is one of the nation's more hazardous occupations, and injury prevention among agricultural workers is a focus of safety and education programs nationwide. This research project investigated the effectiveness of a culturally appropriate body mechanics education program for fruit warehouse workers in Washington State. The purpose of the body mechanics education program was to promote correct ergonomic behavior among migrant and seasonal fruit warehouse workers. Participants received instruction in proper body mechanics by viewing a videotaped Spanish-language theatre program (with English subtitles) followed by a demonstration and practice of correct lifting techniques and selected stretches for injury prevention. A written pre- and post-test to assess body mechanics knowledge and an evaluation of lifting methods were administered at the time of the training and again two weeks later. The results indicated culturally appropriate body mechanics education is an effective intervention for increasing knowledge and promoting correct lifting techniques. However, further research is indicated to examine the significance of supervised and individualized, job specific practice on affecting more lasting changes in work-related body mechanics and lifting behaviors. PMID- 19127018 TI - Outcome of work rehabilitation for people with various disabilities and stability at a one-year follow-up. AB - The main purpose of this study was to investigate whether the outcome obtained at the end of a work rehabilitation programme for people with various disabilities were stable in a one-year perspective. An additional aim was to explore which factors influenced the outcome upon completion of work rehabilitation and at the one-year follow-up. The study used information from a database and comprised 291 clients who had completed rehabilitation at least one year ago. Variables such as sociodemographic data, parameters describing the work rehabilitation period and work capacity upon completion of the programme and at the one-year follow-up were recorded. At the one-year follow-up, the clients also performed self-ratings that comprised three questions: perception of the ability to influence their own situation, the social support available from friends and family, and the need for support from society. The younger clients were more likely to be working after the rehabilitation, but this was not the case at the follow-up. Gender, ethnic origin was not related to outcome directly after rehabilitation, but at the follow-up fewer women than expected were working and more clients than expected of a Scandinavian origin were working. Women rated their need of social support higher than men, and clients from outside Scandinavia perceived a lower ability to influence their own situation. Gender, duration of sick leave, time spent in rehabilitation, and ethnic origin were variables that were of no importance for the outcome of rehabilitation but were of significance for the outcome at the one year follow-up. These results demonstrate the necessity of not only studying the outcome directly after rehabilitation, but also the stability of results at in a follow-up perspective, since the individual's situation changes over time. PMID- 19127019 TI - Economic analyses of rehabilitation services as research methodologies. AB - This article describes the objectives and methods involved in economic analyses of rehabilitation services. Demonstrated approaches to cost-effectiveness and cost-benefit studies are presented, illustrated with examples from the contemporary literature. The strengths and limitations of economic analyses are considered within the context of current trends in rehabilitation policy and practice. PMID- 19127020 TI - Ergonomics for schools. AB - Ergonomics in Schools has focused on micro-ergonomics issues such as mismatch between student body sizes and their desks and chairs, the weight of schoolbags and the prevalence of musculoskeletal disorders amongst school students. Very little of this has been turned into practice. Although advances in this context are being made by the International Ergonomics (IEA) Technical Committee (TC) on Ergonomics for Children in Educational Environments (ECEE), the UK Ergonomics Society Special Interest Group on Ergonomics for Schools and the New Zealand Ergonomics Society Ergonomics/Human factors in Education (EHFIE) Special Interest Group, there has been less focus on macro-ergonomics issues such as learning environments, ergonomics pedagogy and curriculum content/structure. This commentary reviews micro-ergonomics aspects of ;Ergonomics in schools' and argues in favour of a new direction: macro-ergonomics for schools, or educational ergonomics in which ergonomists seek to integrate ergonomics into national educational curricula in concert with Ministries of Education and also via inclusion of ergonomics modules in teacher training. PMID- 19127021 TI - COX2 in CNS neural cells mediates mechanical inflammatory pain hypersensitivity in mice. AB - A cardinal feature of peripheral inflammation is pain. The most common way of managing inflammatory pain is to use nonsteroidal antiinflammatory agents (NSAIDs) that reduce prostanoid production, for example, selective inhibitors of COX2. Prostaglandins produced after induction of COX2 in immune cells in inflamed tissue contribute both to the inflammation itself and to pain hypersensitivity, acting on peripheral terminals of nociceptors. COX2 is also induced after peripheral inflammation in neurons in the CNS, where it aids in developing a central component of inflammatory pain hypersensitivity by increasing neuronal excitation and reducing inhibition. We engineered mice with conditional deletion of Cox2 in neurons and glial cells to determine the relative contribution of peripheral and central COX2 to inflammatory pain hypersensitivity. In these mice, basal nociceptive pain was unchanged, as was the extent of peripheral inflammation, inflammatory thermal pain hypersensitivity, and fever induced by lipopolysaccharide. By contrast, peripheral inflammation-induced COX2 expression in the spinal cord was reduced, and mechanical hypersensitivity after both peripheral soft tissue and periarticular inflammation was abolished. Mechanical pain is a major symptom of most inflammatory conditions, such as postoperative pain and arthritis, and induction of COX2 in neural cells in the CNS seems to contribute to this. PMID- 19127024 TI - Study of the relationship of psychosocial disorders to bruxism in adolescents. AB - BACKGROUND AND AIM: Bruxism has been defined as a diurnal or nocturnal parafunctional habit. Etiology of bruxism has remained controversial and some investigators believe that psychological factors may play a major role in promoting and perpetuating this habit. The aim of this case-control study was to assess the existence of an association between bruxism and psychosocial disorders in adolescents., Participants were chosen among 114, 12-14 year old students (girls). They were divided into two groups, bruxers and nonbruxers, on the basis of both validated clinical criteria and interview with each patient. A few participants were excluded on the basis of presence of systemic disorders, TMJ disorders, other oral habits, primary teeth, defective restorations and premature contacts. Following matching of two groups in regard to parent's age and education, mother's marital status, child support status, mother's employment status, and socio-economical status, 25 cases and 25 controls were enlisted. A self report validated questionnaire (YSR, 11-18 yr) was then filled out by both groups for the evaluation of 12 psychosocial symptoms. RESULTS: Remarkable differences in certain psychosocial aspects were found between the two groups. Prevalence of psychosocial disorders including Thought Disorders (P < 0.005), Conduct Disorders (P < 0.05), Antisocial Disorders (P < 0.06) as identified by YSR was significantly higher in bruxers. Significant differences between the two groups also emerged in total YSR scores (P < 0.005). The results of Odds Ratio revealed that a bruxer adolescent has 16 times greater probability for psychosocial disorders than a non-bruxer one. Fischer exact test and T-test were used and Odds Ratio and Confidence Interval was estimated. CONCLUSION: Support to the existence of an association between bruxism and psychosocial disorders has been provided. PMID- 19127025 TI - Glass fiber-reinforced composite resin as a space maintainer: a clinical study. AB - The aim of this study is to evaluate the use of glass fiber-reinforced composite resin (GFRCR) as a space maintainer and to compare it with a conventional band and-loop space maintainer. A total of 30 children (23 boys and 7 girls) aged 6-8 years were selected for the study. Each of these children required maintenance of space due to premature loss of primary first molars in at least two quadrants. In one quadrant, a GFRCR space maintainer was applied and in the other quadrant a band-and-loop space maintainer was cemented. Patients were recalled at regular intervals over 12 months and retention of both the types of space maintainers was evaluated. The retention of the GFRCR space maintainer was found to be superior to that of the band-and-loop space maintainer, but this difference was not statistically significant. PMID- 19127022 TI - Regulator of G protein signaling 2 mediates cardiac compensation to pressure overload and antihypertrophic effects of PDE5 inhibition in mice. AB - The heart initially compensates for hypertension-mediated pressure overload by enhancing its contractile force and developing hypertrophy without dilation. Gq protein-coupled receptor pathways become activated and can depress function, leading to cardiac failure. Initial adaptation mechanisms to reduce cardiac damage during such stimulation remain largely unknown. Here we have shown that this initial adaptation requires regulator of G protein signaling 2 (RGS2). Mice lacking RGS2 had a normal basal cardiac phenotype, yet responded rapidly to pressure overload, with increased myocardial Gq signaling, marked cardiac hypertrophy and failure, and early mortality. Swimming exercise, which is not accompanied by Gq activation, induced a normal cardiac response, while Rgs2 deletion in Galphaq-overexpressing hearts exacerbated hypertrophy and dilation. In vascular smooth muscle, RGS2 is activated by cGMP-dependent protein kinase (PKG), suppressing Gq-stimulated vascular contraction. In normal mice, but not Rgs2-/- mice, PKG activation by the chronic inhibition of cGMP-selective phosphodiesterase 5 (PDE5) suppressed maladaptive cardiac hypertrophy, inhibiting Gq-coupled stimuli. Importantly, PKG was similarly activated by PDE5 inhibition in myocardium from both genotypes, but PKG plasma membrane translocation was more transient in Rgs2-/- myocytes than in controls and was unaffected by PDE5 inhibition. Thus, RGS2 is required for early myocardial compensation to pressure overload and mediates the initial antihypertrophic and cardioprotective effects of PDE5 inhibitors. PMID- 19127026 TI - Comparative evaluation of shear bond strength of different bracket base designs in dry and wet environments. AB - One hundred and thirty five extracted premolars were selected and bonded with three types of brackets with different bracket base designs viz. Gemini (foil mesh), Miniature twin (Laser etched) and Mini Dynalocks (Integral base). These brackets were bonded with Transbond XT adhesive using two types of primers viz. Conventional Transbond XT in dry conditions and M.I.P in dry and wet environments. The specimens were tested for SBS using Instron machine at C.I.P.E.T. (Amritsar), Punjab. Results were statistically analyzed. RESULTS AND DISCUSSION: The Miniature twin brackets revealed higher SBS values as compared to Gemini and Mini Dynalock in dry and wet conditions. All the groups showed higher SBS values in dry conditions as compared to wet conditions respectively. The Gemini and Mini Dynalock brackets showed very little difference of SBS values. CONCLUSION: Miniature twin brackets with laser etched bracket base in dry and wet environments have revealed the highest SBS values. All the groups have revealed clinically acceptable SBS value of above 8 MPa in moist conditions. PMID- 19127027 TI - Effect of glutaraldehyde and ferric sulfate on shear bond strength of adhesives to primary dentin. AB - AIM: The present study was undertaken to evaluate the effect of alternative pulpotomy agents such as glutaraldehyde and ferric sulfate on the shear bond strength of self-etch adhesive systems to dentin of primary teeth. MATERIALS AND METHODS: Eighty human primary molar teeth were sectioned in a mesiodistal direction and divided into experimental and control groups. Lingual dentin specimens in experimental groups were treated with glutaraldehyde and ferric sulfate. Buccal surfaces soaked in water served as control group. Each group was then divided into two groups based on the adhesive system used: Clearfil SE Bond and Adper Prompt L-Pop. A teflon mold was used to build the composite (Filtek Z 250) cylinders on the dentinal surface of all the specimens. Shear bond strength was tested for all the specimens with an Instron Universal Testing Machine. The failure mode analysis was performed with a Scanning Electron Microscope (SEM). RESULTS: The results revealed that glutaraldehyde and ferric sulfate significantly reduced the shear bond strength of the tested adhesive systems to primary dentin. Clearfil SE Bond showed much higher shear bond strength than Adper Prompt L Pop to primary dentin. SEM analysis revealed a predominant cohesive failure mode for both adhesive systems. CONCLUSION: This study revealed that the pulpotomy medicaments glutaraldehyde and ferric sulfate adversely affected the bonding of self-etch adhesive systems to primary dentin. PMID- 19127028 TI - Distribution of malocclusion types among thumb suckers seeking orthodontic treatment. AB - The present study was conducted to evaluate the influence of thumb sucking and its duration on the type of malocclusion. A total of 410 North Indian individuals between ages 12-30 years, 161 males and 249 females were examined. A specially designed proforma was used to record the detailed history of the individual. Among the etiological factors it was found that history of thumb sucking was present in 13.9% and significantly related to Class II div 1 malocclusion. When thumb sucking exceeded 18 months, it was found statistically significant (P < 0.1) in Class II skeletal malocclusion, in open bite (P < 0.1) and in extreme overjet (P < 0.5). CONCLUSION: If thumb sucking is not treated early, it can cause skeletal Class II malocclusion, anterior open bite and excessive overjet. PMID- 19127029 TI - Griscelli syndrome: a case report of Reye's syndrome and atopic dermatitis history. AB - Griscelli syndrome (GS) is a rare autosomal recessive disorder that results in pigmentary dilution of the skin and the hair (silver hair), with the presence of large clumps of pigment in hair shafts, and an accumulation of melanosomes in melanocytes. Sixty cases of GS have been reported in the literature, but we could find no description of its oro-dental symptoms. Reye's syndrome (RS) is characterized by acute noninflammatory encephalopathy and renal and hepatic failure, while atopic dermatitis (AD) is a skin disorder with an immunologic basis. The aim of this paper is to describe the oro-dental and physical findings in a girl who had been diagnosed with GS at 3.5 years of age; she also had AD as well as a history of RS at infancy. We discuss the possible relationship between the three syndromes. PMID- 19127030 TI - Esthetic and endosurgical management of Turner's hypoplasia; a sequlae of trauma to developing tooth germ. AB - Turner's hypoplasia usually manifests as a portion of missing or diminished enamel, generally affecting one or more permanent teeth in the oral cavity. A case report of 8 year old girl who met with trauma at 2 years of age leading to primary incisors being knocked out, reported after 6 years with complaint of pain and discharge in her anterior malformed teeth is discussed in this article. The permanent incisors erupted with dilacerated crown, root malformations and missing enamel. Further, patient developed sinus, lateral root pathology, tooth mobility and malocclusion in relation to affected teeth which were managed by esthetic, functional, endodontic and surgical procedure. Root canal treatment along with palatal contouring and esthetic restoration by light cure composite was performed on the tooth with crown dilaceration and sinus, where as surgical management was considered for the tooth with root malformation. PMID- 19127031 TI - Osteomyelitis in infantile osteopetrosis: a case report with review of literature. AB - Osteopetrosis is a rare genetic disorder that causes generalized sclerosis of bone due to a defect in bone resorption and remodeling. Dental abnormality may be attributed to pathological changes in bone remodeling. Osteomyelitis is well documented as a complication of osteopetrosis. This disease can be severe and difficult to treat in the osteopetrotic patient. This is a case of 10-year-old girl previously diagnosed as suffering from osteopetrosis and presenting with the complaint of swelling of jaw with extra-oral draining sinus. PMID- 19127032 TI - Integrity, immunity, reactivity, restorativity: biolessons off brain abscess. PMID- 19127033 TI - Platelet glycoprotein IIb/IIIa inhibitors in acute ischemic stroke. AB - Acute ischemic stroke (AIS) is a common cause of morbidity and mortality worldwide. Thrombolytic therapy with tissue plasminogen activator, the only approved treatment for AIS, is received by less than 2% of patients. Moreover, there is a slight increase in hemorrhagic complications with thrombolysis. Therefore, there is a need for newer therapeutic modalities in AIS, which could be used in window periods beyond 3-6 h after stroke onset with fewer hemorrhagic complications. Glycoprotein IIb/IIIa inhibitors (GPI), after their initial success in patients with acute coronary syndromes, promised much in patients with AIS over the past decade or so. However, their exact role in patients with AIS, including the window periods and type of strokes, and the risk of symptomatic or asymptomatic hemorrhage are unclear at the moment. The current review focuses on the literature concerning the use of GPI in AIS and looks at the available evidence regarding their use. Abciximab thought to be safe and effective in initial case series and early trials, has not been shown to improve outcomes in AIS, and is associated with higher rates of hemorrhage. Tirofiban appears to be safe and effective in initial trials and there is a need to conduct further trials to establish its role in AIS. PMID- 19127034 TI - Advanced cerebral monitoring in neurocritical care. AB - New cerebral monitoring techniques allow direct measurement of brain oxygenation and metabolism. Investigation using these new tools has provided additional insight into the understanding of the pathophysiology of acute brain injury and suggested new ways to guide management of secondary brain injury. Studies of focal brain tissue oxygen monitoring have suggested ischemic thresholds in focal regions of brain injury and demonstrated the interrelationship between brain tissue oxygen tension (P bt O 2 ) and other cerebral physiologic and metabolic parameters. Jugular venous oxygen saturation (SjVO 2 ) monitoring may evaluate global brain oxygen delivery and consumption, providing thresholds for detecting brain hypoperfusion and hyperperfusion. Furthermore, critically low values of P bt O 2 and SjVO 2 have also been predictive of mortality and worsened functional outcome, especially after head trauma. Cerebral microdialysis measures the concentrations of extracellular metabolites which may be relevant to cerebral metabolism or ischemia in focal areas of injury. Cerebral blood flow may be measured in the neurointensive care unit using continuous methods such as thermal diffusion and laser Doppler flowmetry. Initial studies have also attempted to correlate findings from advanced neuromonitoring with neuroimaging using dynamic perfusion computed tomography, positron emission tomography, and Xenon computed tomography. Additionally, new methods of data acquisition, storage, and analysis are being developed to address the increasing burden of patient data from neuromonitoring. Advanced informatics techniques such as hierarchical data clustering, generalized linear models, and heat map dendrograms are now being applied to multivariable patient data in order to better develop physiologic patient profiles to improve diagnosis and treatment. PMID- 19127035 TI - Prehospital care of head injured patients. AB - Resuscitation of head injured patients at the accident site is paramount in minimizing morbidity and mortality. This can be achieved through prehospital care which is nonexistent in our country. This review is a step forward, so that we can formulate guidelines in this regard. PMID- 19127036 TI - Diabetic and non-diabetic lumbosacral radiculoplexus neuropathy. AB - BACKGROUND: Lumbosacral radiculoplexus neuropathy (LRPN) originally described in diabetic patients is a distinct clinical condition characterized by debilitating pain, weakness and atrophy most commonly affecting the proximal thigh muscles asymmetrically. The syndrome is usually monophasic and preceded by significant weight loss (at least more than 10 lbs). Though a self-limited condition, recovery is gradual with some residual weakness. Recent advances and research has provided new insights in the pathogenesis and thereby management of this syndrome. In this paper, we review the clinical and diagnostic features as well as discuss recent insights and treatment strategies along with our experience in the management of patients with diabetic and non-diabetic LRPN. MATERIALS AND METHODS: Literature in English published between 1953 and 2008 was searched in the MEDLINE and Pubmed database, maintained by the US National library of medicine and National institutes of health, using key words of diabetic amyotrophy, lumbosacral radiculoplexus neuropathy, diabetic proximal neuropathy, diabetic radiculopathy and diabetic lumbosacral plexopathy. In addition, literature reported in various textbooks on peripheral neuropathy was reviewed as well. OBSERVATION: The diagnosis relies mostly on clinical suspicion and characteristic electromyographic findings. The exact pathogenesis of the illness remains unknown, but there seems to be a component of immune-mediated inflammatory microvasculitis which causes secondary ischemia of the lumbosacral plexus. This has prompted a trial of immunosuppressive agents (like steroids) with an attempt to alter the course of the illness. A few reports have noted that immunosuppression when instituted early in the course of the illness (within three months of symptom onset) may hasten the recovery and improve symptoms. CONCLUSION: Though the exact mechanism of LRPN in diabetic and non-diabetic patients remains unknown, new evidence alludes to an underlying inflammatory vasculitic process. Early treatment with immunosuppressants may be beneficial in some cases, although the data available at this time is limited to a small cohort of patients. The decision is individualized weighing the risks and benefits in a given patient. Future research in this direction with double-blinded case controlled studies is required to investigate this further. PMID- 19127037 TI - Comparison of SF-36 and WHOQOL-100 in patients with stroke. AB - BACKGROUND AND AIMS: Two widely used evaluation tools for the quality of life are the 36-item Short-Form Health Survey (SF-36) and World Health Organization Quality of Life Assessment (100-item version) (WHOQOL-100), however, these tools have not been compared for patients with stroke to date. The specific objectives of this study were: 1) to study the effect of stroke on quality of life (QOL) as measured by the SF-36 and by the WHOQOL-100, and 2) to compare these two instruments. SETTINGS AND DESIGN: Seventy patients who were admitted to the neurology clinic six months after stroke were included in this study. PATIENTS AND METHODS: As a data-collecting device, the SF-36 and WHOQOL-100 scales were used. An additional questionnaire was administered to obtain demographic data. STATISTICAL ANALYSIS: Pearson correlation analysis was performed and Blant-Altman Plots were used. Psychometric analysis was performed. RESULTS: In stroke, the most flustered domains of quality of life were vitality and general health perception fields in the SF-36 and in the WHOQL-100, independence level field, overall QOL and general health perceptions. While there was a fair degree of relationship (r= 0.25-0.50) between general health perceptions, physical, social and mental fields that were similar fields of scales, a fair and moderate to good relationship was found between different fields. Limits of agreement in similar domains of the two instruments were very large. In all four demonstrated Bland Altman plots, there was agreement of the scales in the measurements of similar fields of quality of life. CONCLUSION: This study demonstrated that both the SF 36 and WHOQOL-100 quality of life scales are useful in the practical evaluation of patients with stroke. PMID- 19127038 TI - Clinical features, laboratory data, management and the risk factors that affect the mortality in patients with postoperative meningitis. AB - BACKGROUND: Nosocomial meningitis is a rare complication following neurosurgical procedures and is associated with high morbidity and mortality. AIM: The aim of this study was to describe the clinical characteristics and the risk factors associated with mortality in patients who developed nosocomial meningitis following neurosurgical operations. SETTING AND DESIGN: Tertiary care hospital and an observational study. MATERIALS AND METHODS: The study subjects included 2265 patients who underwent various neurosurgical operations during 2003-05. The diagnosis of nosocomial meningitis was based on the Center for Disease Control criteria. STATISTICAL ANALYSIS: It was performed by using Statistical Package for Social Sciences for Windows 10.0 program. RESULTS: The incidence of postoperative nosocomial meningitis was 2.7% (62 episodes in 49 patients among 2265 patients operated). Staphylococcus aureus and Acinetobacter spp. were the most frequently isolated pathogens. Of the 49 with meningitis 20 (40.8%) patients died. In the logistic regression analysis model, Glascow coma scale score less than 10 (Odds Ratio (OR): 19.419, 95% Confidence Interval (CI); 1.637-230.41, P = 0.001), and low cerebrospinal fluid glucose level (< or = 30 mg/ dL) (OR: 10.272, 95% CI; 1.273-82.854, P = 0.002), and presence of concurrent nosocomial infection (OR: 28.744, 95% CI;1.647-501.73, P =0.001) were the independent risk factors associated with mortality. CONCLUSION: The mortality in patients who developed meningitis was high. The high percentage of concurrent nosocomial infections was associated with a high mortality rate which was a serious problem. PMID- 19127039 TI - Central motor conduction in brachial monomelic amyotrophy. AB - BACKGROUND: Prevalence of subclinical involvement of motor pathways in clinically diagnosed Brachial Monomelic Amyotrophy (BMMA) is unknown. AIMS: To determine the prevalence of subclinical involvement of central motor pathways in BMMA using transcranial magnetic stimulation. SETTING AND DESIGN: Prospective case-control study. MATERIALS AND METHODS: Central motor conduction time (CMCT) was determined by 'F' wave method using figure-of-eight coil attached to Magstim 200 stimulator, in 17 patients with BMMA. Motor evoked potentials were recorded from first dorsal interosseous of the affected (AFF) and unaffected upper limbs (UNAFF) at rest and during partial contraction. Comparison was made with data from 10 healthy controls (CTRL). STATISTICAL ANALYSIS: Descriptive analysis and Analysis of Variance (ANOVA). RESULTS: Compared to controls, the mean CMCT of AFF was significantly prolonged, both at rest and contraction: (a) Rest: AFF-6.68+/-1.78 ms, UNAFF-6.36+/-1.16 ms, CTRL-5.71+/-1.02 ms; Fisher's PLSD for AFF vs. CTRL: P =0.037, (b) Contraction: AFF-5.78+/-1.62 ms, UAFF - 4.86+/-1.38 ms, CTRL-4.06+/ 0.80 ms; Fisher's PLSD for AFF vs. CTRL; P =0.0002, AFF vs. UNAFF- P =0.044). Prolonged CMCT (>mean+2SD of controls) was observed in 29.4% of AFF and 6.25% of UNAFF at rest, and in 47.1% and 23.5% respectively during contraction. CONCLUSIONS: Dysfunction of central motor pathways was observed in both affected and unaffected upper limbs of some patients with BMMA of upper limbs. The dysfunction was more pronounced during voluntary contraction. A larger study is needed to validate the significance of these findings. PMID- 19127040 TI - Cryptococcal meningitis in HIV infected: experience from a North Indian tertiary center. AB - BACKGROUND: Cryptococcal meningitis is a common opportunistic infection in Human Immunodeficiency Virus (HIV)-infected individuals. There is little information specifically addressing cryptococcal meningitis in HIV-infected patients from North India. AIMS: To determine clinical presentation, hospital course, response to treatment, complications developed, in-hospital mortality, any recurrence of cryptococcal meningitis and reasons of recurrence during follow-up. SETTINGS AND DESIGN: A retrospective observational study undertaken in a large tertiary care center. PATIENTS AND METHODS: Patient's demographic data, presenting clinical symptomatology, physical findings, laboratory parameters, cerebrospinal fluid (CSF) examination findings, side-effects of treatment, development of any complications and hospital outcome were analyzed. During follow-up any recurrence of cryptococcal meningitis, possible reasons of recurrence, type of treatment received, complications developed and outcome was recorded as well. RESULTS: Forty patients diagnosed to have cryptococcal meningitis were analyzed. Twenty two (55%) patients had acute/ subacute presentation. Thirty-six (90%) patients presented with headache and 18 (45%) had altered sensorium. Twenty (50%) patients had no cells in the CSF. Hypoglycorrhchia was seen in 30 (75%) patients. Cryptococcal meningitis was the first acquired immune deficiency syndrome (AIDS) defining illness in 30 (75%) patients. Thirty-five patients developed some adverse effects to amphotericin-B. Thirty-three patients improved with treatment while three patients died. Four patients had recurrence of cryptococcal meningitis within six months of first episode. Non-compliance of fluconazole therapy was the reason for recurrence in all of these patients. CONCLUSIONS: Cryptococcal meningitis is a common initial AIDS-defining illness. Acute and/or subacute presentation of cryptococcal meningitis is not uncommon in HIV-infected individuals. An early diagnosis of HIV infection might reduce the incidence of this infection. PMID- 19127041 TI - Proprioceptive loss in leprous neuropathy: a study of 19 patients. AB - BACKGROUND: Leprosy presents commonly with mononeuritis multiplex, affecting mainly the exteroceptive sensations. Neuropathy with a significant afferent large fiber element is considered to be an uncommon manifestation of leprous neuropathy. AIMS: To evaluate the clinical and neurophysiologic aspects of a subset of patients with leprous neuropathy having clinical proprioceptive loss. SETTINGS AND DESIGN: Prospective study of patients with a diagnosis of peripheral neuropathy secondary to leprosy having proprioceptive loss. MATERIALS AND METHODS: Consecutive patients seen during a two-year period (2004 and 2005) diagnosed to have leprous neuropathy with proprioceptive abnormalities on clinical examination were included. The diagnosis of leprosy was achieved by clinical features along with positive skin biopsy, split skin smears or nerve biopsy. Their clinical and electrophysiological characteristics were studied. STATISTICAL METHODS: The results were analyzed using Chi-Square test. Values less than 0.05 were considered to be statistically significant . RESULTS AND CONCLUSIONS: We observed predominance (68.42%) of multibacillary of leprosy. Symmetrical neuropathies outnumbered mononeuritis multiplex (12:7). The pan sensory neuropathy had a mean duration of 24.32 months, but sometimes appeared early in the course of the disease. Areflexia and electrophysiological evidence of proximal affection was common, reflecting proximal spread of neuropathic process. Such patients have a higher incidence of developing deformities and ulcerations and they represent a vulnerable subset of patients with leprosy. PMID- 19127042 TI - Epidermal growth factor receptor and proliferating cell nuclear antigen in astrocytomas. AB - AIMS: The involvement of various growth factors, growth factor receptors and proliferative markers in the molecular pathogenesis of astrocytic neoplasms are being studied extensively. Epidermal Growth Factor Receptor (EGFR) gene overexpression occurs in nearly 50% of cases of glioblastoma. Since EGFR and proliferating cell nuclear antigen (PCNA) are involved in mitogenic signal transduction and cellular proliferation pathway, we have studied the correlation between the expression of EGFR and PCNA labeling index in astrocytic tumors. MATERIALS AND METHODS: We investigated the immunohistochemical expression of EGFR and PCNA using the appropriate monoclonal antibodies in 40 cases of astrocytic tumors of which 21 cases were glioblastoma, eight cases were Grade III or anaplastic astrocytomas and six cases were Grade II or diffuse astrocytomas and five cases were Grade I or pilocytic astrocytomas. RESULTS: Both the EGFR expression and PCNA labeling index increase with increasing grades of astrocytomas with a significantly high percentage of cells showing positive staining for both EGFR and PCNA in GBM and Grade III astrocytomas compared to Grade II astrocytomas. The expression levels of both EGFR and PCNA were low in Grade I or pilocytic astrocytomas. CONCLUSIONS: A significant correlation was found between EGFR overexpression and PCNA labeling index in Grade III and Grade II astrocytomas and glioblastoma. These suggest that the tumor proliferation, at least in higher grades of astrocytomas is dependent in some measure on EGF and EGFR-related signaling pathways. PMID- 19127043 TI - Dysphagia in acute stroke: correlation with stroke subtype, vascular territory and in-hospital respiratory morbidity and mortality. AB - AIMS: The study aimed at correlation of post-stroke dysphagia with area and volume of infarct/ bleed, and with subsequent in-hospital respiratory morbidity and mortality. MATERIALS AND METHODS: 50 patients of acute stroke were serially recruited. Standard Staff swallowing assessment was performed within 24 hours of admission along with pulse oximetry. Ischemic strokes were classified as per OCPS registry. In-hospital respiratory morbidity and mortality, mode of nutrition and disability status at discharge were noted. RESULTS: 21/50 (42%) patients had post stroke dysphagia during their hospital course. Among infarcts, Total Anterior Circulation Infarcts (TACI) had 100% incidence of dysphagia, followed by Partial Anterior Circulation Infarcts (PACI-36%), Posterior Circulation infarcts (POCI 33%), and Lacunar infarcts (LACI-18%). 67% of hemorrhages had post-stroke dysphagia. Staff swallowing assessment had a sensitivity and specificity of 75% and 73% respy., for predicting respiratory morbidity. The corresponding figures for Pulse oximetry were 79% and 91%. PMID- 19127044 TI - A case of tactile agnosia with a lesion restricted to the post-central gyrus. AB - Tactile agnosia has been described after lesions of the primary sensory cortex but the exact location and extension of those lesions is not clear. We report the clinical features and imaging findings in a patient with an acute ischemic stroke restricted to the primary sensory area (S1). A 73-year-old man had a sudden onset of a left alien hand, without left hemiparesis. Neurological examination showed intact primary sensory functions, but impaired recognition of shape, size (macrogeometrical) and texture (microgeometrical) of objects; damage confined to the post-central gyrus, sparing the posterior parietal cortex was demonstrated on MRI. An embolic occlusion of the anterior parietal artery was suspected as mechanism of stroke. Tactile agnosia with impaired microgeometrical and macrogeometrical features' recognition can result from a single lesion in the primary sensory cortex (S1) in the right parietal hemisphere, sparing other regions of the cerebral cortex which presumably participate in tactile object recognition. PMID- 19127045 TI - Hypersexuality following subthalamic nucleus stimulation for Parkinson's disease. AB - Subthalamic nucleus (STN) stimulation is an established surgical treatment for Parkinson's disease (PD). Though the motor benefits of STN stimulation are well understood, its cognitive and behavioral effects are still not fully understood. Manic psychosis, hypersexuality, pathological gambling and mood swings are associated with advanced PD. There have been reports to suggest improvement or worsening in these symptoms following STN deep brain stimulation (DBS). We report two cases as the sole behavioral side-effects of STN stimulation despite good clinical improvement on long-term follow-up. These patients and literature review suggests the complex role of STN stimulation in motor and behavioral control. PMID- 19127046 TI - Congenital irreducible atlantoaxial dislocation associated with cervical intramedullary astrocytoma causing progressive spastic quadriparesis. AB - Simultaneous presence of congenital irreducible atlantoaxial dislocation (AAD) and cervical intramedullary astrocytoma has not been previously described and may cause disabling myelopathy. This 55-year-old lady presented with suboccipital pain, spastic quadriparesis, Lhermitte's phenomenon and sphincteric disturbances. Lateral radiographs and magnetic resonance imaging showed irreducible AAD, occipitalized atlas, C2-3 fusion, and,an intramedullary tumor from C2-5 level iso to-hypointense, non-enhancing, except in a small segment in the dorsal C2 level. A suboccipital craniectomy with C2-5 laminectomy revealed a greyish-white tenacious tumor. The tumor was decompressed using a C2-5 midline myelotomy and duroplasty. An occipitocervical lateral mass fixation was performed. Histopathology revealed a low-grade astrocytoma. At three-month follow-up, her spasticity had decreased and quadriparesis and sphincteric disturbances were persisting. Postoperative lateral radiographs and intrathecal contrast CT scan showed a stable occipitocervical construct. Thus, the suboccipital craniectomy and laminectomy with midline myelotomy and duroplasty facilitated space for progressively expanding intramedullary astrocytoma with irreducible AAD; the lateral mass fixation provided stability at the craniovertebral junction. PMID- 19127047 TI - Foramen-fallacy: descriptive-delusion. PMID- 19127048 TI - Familial mirror movements over five generations. PMID- 19127050 TI - Acute lithium toxicity: two cases with different outcomes. PMID- 19127049 TI - Unilobar multiple cerebral hydatid cyst: a rare disease. PMID- 19127051 TI - MRI diagnosis of neurolymphomatosis of the brachial plexus. PMID- 19127052 TI - Anesthetic management of a patient with Huntington's chorea. PMID- 19127053 TI - Infraoptic azygous anterior cerebral artery. PMID- 19127054 TI - Endovascular management of a refractory traumatic aneurysm of the internal carotid artery using a covered stent graft. PMID- 19127055 TI - Cushing's disease presenting as an intracerebral hemorrhage. PMID- 19127056 TI - Duplication of the common carotid artery and the ipsilateral vertebral artery with a fenestration of the contralateral common carotid artery. PMID- 19127057 TI - Cytomegalovirus polyradiculopathy: a rare neurological manifestation of acquired immunodeficiency syndrome. PMID- 19127058 TI - Lhermitte-Duclos disease. PMID- 19127059 TI - Monostotic fibrous dysplasia of the C4 cervical spine. PMID- 19127062 TI - Oxidative imbalance in nonstimulated X-adrenoleukodystrophy-derived lymphoblasts. AB - X-Adrenoleukodystrophy (X-ALD) is a peroxisomal disorder characterized by accumulation of very-long-chain (VLC) fatty acids, which induces inflammatory disease and alterations in cellular redox, both of which are reported to play a role in the pathogenesis of the severe form of the disease (childhood cerebral ALD). Here, we report on the status of oxidative stress (NADPH oxidase activity) and inflammatory mediators in an X-ALD lymphoblast cell line under nonstimulated conditions. X-ALD lymphoblasts contain nearly 7 times higher levels of the C(26:0) fatty acid compared to controls; these levels were downregulated by treatment with sodium phenylacetate (NaPA), lovastatin or the combination of both drugs. In addition, free-radicals synthesis was elevated in X-ALD lymphoblasts, and protein levels of the NADPH oxidase gp91(PHOX) membrane subunit were significantly upregulated, but no changes were observed in the p47(PHOX) and p67(PHOX) cytoplasmic subunits. Unexpectedly, there was no increase in gp91(PHOX) mRNA levels in X-ALD lymphoblasts. Furthermore, X-ALD lymphoblasts produced higher levels of nitric oxide (NO) and cytokines (tumor necrosis factor-alpha and interleukin 1 beta), and treatment with NaPA or lovastatin decreased the synthesis of NO. Our data indicate that X-ALD lymphoblasts are significantly affected by the accumulation of VLC fatty acids, which induces changes in the cell membrane properties/functions that may, in turn, play a role in the development/progression of the pathogenesis of X-ALD disease. PMID- 19127064 TI - Peanut allergy diagnosis in the context of grass pollen sensitization for 125 patients: roles of peanut and cross-reactive carbohydrate determinants specific IgE. AB - BACKGROUND: In vitro testing for food allergy may yield clinically irrelevant results due to cross-reactive carbohydrate determinants (CCD) specific immunoglobulin E (sIgE) induced by pollen exposure. The performances of 2 in vitro methods were evaluated for peanut sIgE measurement in patients allergic to grass pollen with or without subsequent allergy to peanuts. The correlation between clinically irrelevant peanut sIgE and the presence of CCD sIgE was investigated. METHODS: In vitro measurement of peanut sIgE was performed using the Pharmacia ImmunoCap system Radio Immuno Assay (RIA) and the Immulite 2000 3gAllergy system. Discrepancies between in vitro results and peanut allergy diagnosis were evaluated by measurement of CCD sIgE using bromelain and ascorbic acid oxydase (AAO). RESULTS: The sensitivity was 100% with both systems for the diagnosis of allergy to peanut (58 patients), nevertheless the specificity obtained with Immulite (73%) was better than that obtained using ImmunoCap (46%) in patients who were not allergic to peanuts, but who had a grass pollen allergy (n = 41). In 22 out of 41 patients who presented clinically irrelevant peanut sIgE results using ImmunoCAP, CCD sIgE was detected in 72% of the cases by bromelain and in 86% by AAO. In 11 patients out of 41 who presented irrelevant peanut sIgE results using Immulite, CCD sIgE was detected in 81% of the cases by bromelain and in 100% by AAO. CONCLUSION: The Immulite 2000 system had better specificity than the ImmunoCap system for accurate diagnosis of peanut allergy in patients allergic to grass pollen. CCD sIgE was identified in most of the false positive peanut sIgE results. PMID- 19127063 TI - Low levels of corticotropin-releasing hormone during early pregnancy are associated with precocious maturation of the human fetus. AB - Elevation in placental corticotropin-releasing hormone (pCRH) during the last trimester of pregnancy has been associated with an increased risk for preterm delivery. Less is known about the consequences for the human fetus exposed to high levels of pCRH early in pregnancy. pCRH levels were measured in 138 pregnant women at least once at 15, 20 and 25 weeks of gestation. At 25 weeks of gestation, fetal heart rate (FHR) responses to a startling vibroacoustic stimulus (VAS) were recorded as an index of maturity. pCRH levels at 15 weeks of gestation, but at no later point, predicted FHR responses to the VAS. Fetuses exposed to the lowest concentrations of pCRH at 15 weeks of gestation exhibited a distinguishable response to the VAS, whereas fetuses exposed to higher levels of pCRH did not respond. The findings suggest that exposure to low levels of pCRH early in gestation may be optimal and associated with a response pattern indicating greater maturity. PMID- 19127066 TI - Atopic dermatitis, dry skin and serum IgE in children in a community in japan. AB - BACKGROUND: Long-term, prospective studies investigating the prevalence, serum IgE and the natural history of atopic dermatitis (AD) in a community are lacking. METHODS: In a Japanese community, the skin of primary school children and junior high school students was examined and a questionnaire was given to their parents; their serum total IgE, and house dust mite (HDM)- and Japanese cedar pollen (JCP) specific IgE levels were also assessed once a year for 9 years. RESULTS: The median AD prevalence in all students (492 in 1998 and 380 in 2004) was 7.6% (6.1 10.4%). The prevalence and the area of skin eruptions of AD decreased with growth. Serum total and HDM-specific IgE levels were high in AD patients, and significant differences were noted for both levels between children with and without later remission of skin eruptions at the time of primary school entry. IgE level increases were noted in the following order: healthy skin < dry skin < AD. In children presenting only with dry skin without atopic disorders, such as AD, asthma and allergic rhinitis, levels of total HDM- and JCP-specific IgE were significantly higher than in children with healthy skin. CONCLUSIONS: The infantile IgE level serves as a prognostic index, and sensitization to inhalant allergens may be easily established in individuals with clinically dry skin, even when AD is not present, and this may lead to the development of atopic disorders. PMID- 19127065 TI - Enhanced interferon-gamma gene expression in T Cells and reduced ovalbumin dependent lung eosinophilia in hypoxia-inducible factor-1-alpha-deficient mice. AB - BACKGROUND: There is growing evidence that hypoxia-inducible transcription factors are involved in the pathophysiology of asthma. Hypoxia-inducible factor 1alpha (HIF-1alpha) in particular controls the expression of many hypoxia regulated genes, but whether HIF-1alpha directly contributes to allergen-driven immune responses is not known. METHODS: Partially HIF-1alpha-deficient mice (HIF 1alpha(+/-)) or wild-type littermate controls were used in all experiments. Spleen CD4+ T cells were stimulated with anti-CD3 plus anti-CD28 antibodies and cytokine secretion was measured in vitro. Mice were sensitized by intraperitoneal injection of ovalbumin (Ova) plus alum, and then challenged by intranasal Ova followed by bronchoalveolar lavage (BAL) and isolation of spleen cells. BAL cells were counted and the differential determined using cytospin, and splenocytes were incubated with Ova to measure recall cytokine production. RESULTS: Interferon gamma secretion was significantly higher in anti-CD3 plus anti-CD28 stimulated CD4+ T cells obtained from HIF-1alpha(+/-) mice compared to wild-type controls. HIF-1alpha(+/-) mice were protected from lung eosinophilia 72 h after allergen challenge, in association with enhanced secretion of interferon-gamma in recall responses of splenocytes. CONCLUSIONS: HIF-1alpha contributes to allergic immune responses and lung eosinophilia in a mouse model of asthma. PMID- 19127067 TI - Co-delivery of ovalbumin and CpG motifs into microparticles protected sensitized mice from anaphylaxis. AB - BACKGROUND: Two major drawbacks of current subcutaneous specific immunotherapy are the risk to induce severe anaphylactic reactions and the need of multiple injections of the allergen to reduce IgE-mediated hypersensitivity. The sustained release of allergens over time provided by poly(lactide-co-glycolide) microparticles (MP) could mimic the current therapeutic schedule and decrease their allergenicity. Moreover, MP could also co-deliver Th1 immunoadjuvants, such as CpG motifs. METHODS: Ovalbumin (OVA)-sensitized BALB/c mice were treated intradermally with OVA or OVA plus CpG containing MP. OVA-specific IgG1 and IgG2a as well as IgE total levels and cytokine production were assessed throughout the experiment. The protection exerted by the MP against allergen challenge was estimated with body temperature changes, mortality and other symptoms. RESULTS: Microparticulated treatments, irrespective of the presence of CpG motifs, elicited a lower IgE/IgG2a ratio than those induced by the allergen in solution (free or with adjuvants). However, after induction of the anaphylactic shock, only mice treated with MP co-encapsulating OVA plus CpG showed a significant lower decrease in body temperature and were totally protected from death. Mice that were injected with OVA plus CpG in solution or with Alum displayed a marked fall of temperature accompanied by high mortality rates (70-100%). CONCLUSION: MP encapsulating both OVA, as an allergen model, and CpG sequences, as a pro-Th1 adjuvant, decreased the risk for OVA sensitization (IgE induction) and protected sensitized mice from anaphylactic shock after allergen provocation. Therefore, the combination of allergens and CpG sequences into MP could perform a safer alternative to current specific immunotherapy. PMID- 19127068 TI - Pimecrolimus reduces eosinophil activation associated with calcium mobilization. AB - BACKGROUND: Pimecrolimus is a calcineurin inhibitor that inhibits T cell and mast cell activation and effectively treats atopic dermatitis. However, its effects on eosinophils, a cell type implicated in allergic disease pathology, are unknown. Therefore, we examined the effects of pimecrolimus on eosinophil superoxide anion production, degranulation and survival. METHODS: Purified eosinophils from normal or atopic donors were incubated with serial dilutions of pimecrolimus (microM to nM) and then stimulated with platelet activating factor (PAF), interleukin 5 (IL5), secretory immunoglobulin A (sIgA) or Alternaria alternata (Alt) fungus extract. Eosinophil activation was monitored by cytochrome c reduction resulting from superoxide anion production and by a 2-site immunoassay for eosinophil derived neurotoxin (EDN) in cellular supernatants, as a marker of degranulation. Eosinophil survival was measured by propidium iodide exclusion using flow cytometry after 4 days in culture. RESULTS: Normal and atopic eosinophil superoxide anion production induced by PAF, and associated with increased intracellular calcium, was inhibited up to 37% with 1 microM pimecrolimus. However, superoxide anion production induced by IL5 and sIgA was not consistently inhibited. EDN release, which ultimately depends on calcium, was inhibited about 30% with PAF, IL5 and sIgA stimulation for normal and atopic donor eosinophils. Furthermore, calcium-dependent Alt-induced EDN release was inhibited up to 49% with nanomolar pimecrolimus. Finally, increased eosinophil survival promoted by IL5 and sIgA was not influenced by pimecrolimus. CONCLUSION: Pimecrolimus moderately inhibits eosinophil superoxide anion production and EDN release associated with calcium mobilization, which may contribute to its efficacy in treating atopic dermatitis. PMID- 19127069 TI - Increases in urinary 9alpha,11beta-prostaglandin f2 indicate mast cell activation in wine-induced asthma. AB - BACKGROUND: Wine-induced asthmatic symptoms may be caused by sulphite additives. Prostaglandin (PG)D2 and cysteinyl leukotrienes (cysLT) are important mediators of asthmatic responses. To determine whether the sulphite additives in wine alter the production of PGD2 and cysLT, asthmatic patients with compelling histories of wine sensitivity were challenged with high- and low-sulphite wines; the urinary metabolites of PGD2 and cysLT were measured before and after challenge. METHODS: Eight self-reporting wine-sensitive asthmatic patients completed double-blind challenges with high- and low-sulphite wines on separate days. Urine samples were collected before and after consumption of 150 ml of wine. Urinary concentrations of 9alpha,11beta-PGF2 and leukotriene (LT)E4 were measured by enzyme immunoassay. RESULTS: Urinary 9alpha,11beta-PGF2 concentrations increased in all subjects following challenge with high-sulphite wine, and the median concentration increased 1.6-fold (p < 0.01). Urinary 9alpha,11beta-PGF2 also increased 1.5-fold after low-sulphite wine challenge, although this did not reach statistical significance (p = 0.08). The median difference in 9alpha,11beta-PGF2 concentration after high-sulphite wine challenge was not significantly different compared with that after low-sulphite wine challenge. Median urinary LTE4 concentrations did not change significantly after either wine challenge. CONCLUSIONS: Increased urinary 9alpha,11beta-PGF2 concentrations following wine challenge suggest mast cell activation as a possible mechanism for wine-induced asthma, although this did not appear to be related to the sulphite additives in wine. Urinary 9alpha,11beta-PGF2 may warrant further assessment as a potential biomarker of reactivity to wine in asthmatic subjects. PMID- 19127070 TI - Association between obesity and atopy in Chinese schoolchildren. AB - BACKGROUND: Despite parallel increases in asthma and obesity prevalence, there is little data on obesity as a risk factor for atopy. The latter is an important phenotype in asthmatic patients. This study investigates the association between asthma traits, atopy and obesity-related markers in Chinese adolescents. METHODS: 486 schoolchildren were recruited among participants of our population-based study on the epidemiology of obesity, and their allergy status was ascertained using a standardized questionnaire. Subjects' anthropometry was recorded on-site, and fasting blood was collected for allergen-specific immunoglobulin E (IgE), lipids and systemic inflammatory biomarkers. RESULTS: 98 (20.2%) subjects were classified as overweight or obese. Obesity status was not associated with asthma, allergic rhinitis or eczema (p > 0.25). Atopy was not associated with age adjusted body mass index (BMI) or waist circumference. Atopy and presence of allergen-specific IgE did not differ between overweight or obese children and those with normal BMI (p > 0.25), although subgroup analysis suggested that cockroach sensitization was more common among males who were obese or overweight (p = 0.045). White cell count (WCC) was higher among atopic than nonatopic children (mean values 6.5 x 10(9)/l vs. 6.2 x 10(9)/l, p = 0.006). Logistic regression revealed WCC to be the only risk factor for atopy (OR 18.97, p = 0.004). CONCLUSIONS: Obesity is not associated with asthma or atopy in Chinese children. High WCC is an important risk factor for atopy in both males and females. Gender does not exert any consistent effect on the association between obesity and allergen sensitization in children. PMID- 19127071 TI - Validation study of the OHIO Chamber in patients with Japanese cedar pollinosis. AB - BACKGROUND: An artificial exposure chamber (OHIO Chamber), which allows dispersal of a fixed concentration of Japanese cedar (JC) pollen under stable conditions, was constructed. This study was conducted to identify the exposure conditions assuring validity of the clinical tests conducted using this chamber. METHODS: Twenty-four adult patients with JC pollinosis were exposed to different concentrations of JC pollen: 0 (only during the summer period), 4,000, 8,000 and 12,000 grains/m3, and the nasal and ocular symptoms were self-assessed during a 4 hour period of exposure. The amount of nasal discharge was measured and the sneezing frequency was recorded. This study was conducted twice during the summer and winter periods, i.e. non-pollen seasons. The reproducibility of the symptoms between the two seasons was assessed. RESULTS: None of the subjects developed any symptom at the pollen concentration of 0 grains/m3. No significant differences in the time to the onset of symptoms were found between the summer and winter study, regardless of the pollen concentration. There were no significant differences between the summer and winter study in the total symptom score and total nasal symptom score at any pollen concentration, suggesting the very favorable reproducibility of symptoms. CONCLUSIONS: Efficient and reproducible results are obtained in patients exposed to JC pollen in the OHIO Chamber. The results suggest the conditions of JC pollen exposure have scientific validity and the OHIO Chamber has the potential to contribute significantly to basic and clinical studies of JC pollinosis. PMID- 19127072 TI - Montelukast is not effective in controlling allergic symptoms outside the airways: a randomised double-blind placebo-controlled crossover study. AB - BACKGROUND: Subjects with atopic syndrome often perceive symptoms from various organs. A single drug that acts on all the syndrome's manifestations would be the ideal treatment. The role of montelukast, a cysteinyl-leukotriene receptor antagonist, is established in treating allergic rhinitis and asthma, but its ability to alleviate atopic symptoms outside the airways is controversial. Our aim was to assess if montelukast could be used to treat all the various symptoms seen in subjects with atopic syndrome. METHODS: A randomised, double-blind, placebo-controlled crossover study on the effect of montelukast in atopic syndrome was conducted during the 2007 pollen season. Forty-five pollen sensitised subjects who had allergic symptoms from both the upper and lower airways and allergic symptoms outside the airways (conjunctivitis, oral symptoms, eczema and/or urticaria) were recruited. The primary outcome parameter was the allergic symptoms, which were assessed using a questionnaire. Secondary outcome parameters were lower-airway inflammation (exhaled nitric oxide) and the need for rescue medication (inhaled beta2-agonists and oral antihistamines). RESULTS: There were no differences between montelukast and placebo treatments in allergic symptoms, in exhaled NO concentration or in the need for oral antihistamines. The need for inhaled beta2-agonists was significantly lower during montelukast treatment. CONCLUSIONS: Montelukast was not effective in treating allergic symptoms outside the airways in subjects suffering from different manifestations of the atopic syndrome. Based on the current results, montelukast should not be recommended as a general drug to treat all the symptoms of atopic syndrome, but it should be considered as a drug for asthma and rhinitis. PMID- 19127073 TI - Nasal challenge with LPS stimulates the release of macrophage inflammatory protein 1alpha. AB - BACKGROUND: Bacterial infections can cause a variety of airway diseases. Toll like receptors (TLRs) directly respond to the presence of microbes and partake in the innate immune defense. TLR4 is activated by lipopolysaccharide (LPS), and has been detected in sinonasal tissue, epithelial cells and various inflammatory cells. Macrophage inflammatory protein 1alpha (MIP-1alpha) is a chemokine released during the inflammatory process. The present study investigated the potential role and regulation of MIP-1alpha in LPS-induced nasal inflammation. METHODS: Thirty-two healthy individuals were intranasally challenged with LPS or vehicle. Nasal lavage was performed, followed by a nasal biopsy. Inflammatory cells were counted, MIP-1alpha levels analyzed and expression of MIP-1alpha mRNA in biopsies quantified. Neutrophils isolated from peripheral blood were treated with LPS and effects on MIP-1alpha release, cell survival, and the involved signal pathways, were investigated. RESULTS: LPS challenge caused an increase of MIP-1alpha in nasal lavage. No corresponding change in mRNA expression was seen in nasal biopsies, suggesting the increase was not due to epithelial synthesis. Neutrophil numbers increased after LPS provocation. Treatment of isolated neutrophils with LPS delayed neutrophil apoptosis and resulted in a time- and concentration-dependent release of MIP-1alpha, which was reduced by inhibitors of transcription and of nuclear factor (NF)-kappaB, protein kinase C (PKC) and p38 MAPK pathways. CONCLUSIONS: Nasal LPS challenge results in release of MIP-1alpha. The release most likely originates from recruited neutrophils, via NF-kappaB-, PKC- and p38 MAPK-dependent pathways. LPS stimulation delayed neutrophil apoptosis. MIP-1alpha may constitute an important mediator in neutrophilic airway disease. PMID- 19127074 TI - Classification of hypersensitivity pneumonitis: a hypothesis. AB - BACKGROUND: Regardless of the causative antigen, hypersensitivity pneumonitis (HP) is usually classified as 'acute', 'subacute' or 'chronic'. Considerable confusion still surrounds this classification because there are no widely accepted criteria to distinguish the various stages. The objective of this study was to determine whether the current classification of HP truly reflects categories of patients with distinct clinical features. METHODS: Data obtained from a large prospective multicenter cohort study (the HP Study) were used to divide a cohort of patients with HP into a limited number of categories (clusters) with maximally differing clinical patterns, without prejudgment. The results of this cluster analysis were compared with the current classification of HP (acute, subacute or chronic). RESULTS: 168 patients were included in the analysis. A 2-cluster solution best fitted the data. Patients in cluster 1 (41 patients) had more recurrent systemic symptoms (chills and body aches) and normal chest radiographs than those in cluster 2 (127 patients) who showed significantly more clubbing, hypoxemia, restrictive patterns on pulmonary function tests and fibrosis on high-resolution computed tomography (HRCT). All p values were <0.0001, using Fisher's exact test. Nodular opacities were seen on HRCT as often in cluster 1 as in cluster 2. There was considerable disagreement between the current classification of HP and the results of our analysis. CONCLUSION: The current classification of acute, subacute and chronic HP is not supported by our analysis. Subacute HP is particularly difficult to define. PMID- 19127075 TI - Analysis of signaling pathways involved in peptidoglycan-induced RANTES production from murine Langerhans cells. AB - BACKGROUND: Our previous study showed that percutaneous application of peptidoglycan from Staphylococcus aureus induced eosinophil infiltration in murine skin through production of CCL5/RANTES (regulated upon activation in normal T cells expressed and secreted) from epidermal Langerhans cells. Although it is well known that peptidoglycan is an agonist of Toll-like receptor (TLR)-2, it is unclear whether other TLR agonists are able to induce RANTES production from Langerhans cells. METHODS: Langerhans cells were purified from murine epidermal cells by the panning method using anti-IA(d) monoclonal antibody. RANTES production by Langerhans cells was investigated by RT-PCR and ELISA. Analysis of the signaling pathways responsible for RANTES production by Langerhans cells was performed by ELISA using N-acetyl-L-cysteine, SP600125, SB203580 and PD98059, which are specific inhibitors of NF-kappaB activation, JNK, p38 MAPK and ERK, respectively, and was finally confirmed by Western blot analysis. RESULTS: Peptidoglycan, poly(I:C), lipopolysaccharide and CpG DNA, which signal through TLR-2, TLR-3, TLR-4 and TLR-9, respectively, were found to strongly induce RANTES production. Treatment with N-acetyl-L-cysteine inhibited all TLR agonist-induced RANTES production. However, treatment with SP600125 did not inhibit CpG DNA-induced RANTES production. Furthermore, treatment with SB203580 inhibited only peptidoglycan- and lipopolysaccharide-induced RANTES production and the inhibition was correlated with that of p38 MAPK phosphorylation. CONCLUSION: These results suggest that the signaling pathway of RANTES production from murine Langerhans cells induced by different TLR stimuli is not necessarily the same, and that inhibition of p38 MAPK may be a more specific therapeutic target for eosinophilic inflammation in patients with atopic dermatitis associated with S. aureus colonization. PMID- 19127076 TI - Carbamazepine hypersensitivity syndrome triggered by a human herpes virus reactivation in a genetically predisposed patient. AB - A case of severe hypersensitivity syndrome, triggered by carbamazepine in the presence of a concomitant active human herpes virus (HHV) 6 and 7 infection is described. To further understand the molecular mechanism of this adverse reaction, analyses of the genetic variants of human leukocyte antigen (HLA) and of the epoxide hydrolase gene (EPHX1), previously associated with carbamazepine hypersensitivity, were performed. A lymphocyte transformation test (LTT) was conducted in order to detect drug-specific lymphocytes. In the hypersensitive patient, 2 genetic factors previously associated with intolerance to carbamazepine were detected: the allele HLA-A*3101 and homozygosity for the variant allele of SNP rs1051740 in EPHX1. Drug-specific lymphocytes could be detected by LTT when the HHV was active (positive PCR for viral DNA and increased anti-HHV 6 IgG titer), but not when it was no longer active. In conclusion, we document a case of severe carbamazepine hypersensitivity triggered by viral reactivation in a patient presenting the interaction of 2 unfavorable genetic factors. PMID- 19127077 TI - Desensitization in interferon-beta1a allergy: a case report. AB - We report a 41-year-old patient with multiple sclerosis (MS) who was successfully desensitized after she developed non-injection-site urticaria and angioedema due to interferon (IFN)-beta1a. Although a few cases of urticaria and anaphylaxis have been reported in the literature, to our knowledge this is the first report of a successful desensitization with IFN-beta1a. Desensitization with IFN-beta1a allowed us to continue with the administration of interferon-beta, which is a mainstay in treatment for MS. PMID- 19127078 TI - Influence of occult hepatitis B virus coinfection on the incidence of fibrosis and hepatocellular carcinoma in chronic hepatitis C. AB - We examined prospectively the influence of occult hepatitis B virus (HBV) infection on the histopathological features and clinical outcome of HCV RNA positive chronic hepatitis (CH-C) and detected hepatitis B core (HBc) particles in hepatocytes. The subjects were 468 patients with CH-C or liver cirrhosis (LC) who were negative for serum hepatitis B surface antigen (HBsAg) by enzyme-linked immunosorbent assay. HBV DNA was detected in serum by nested PCR. HBsAg and HBc antigen (HBcAg) in liver were investigated using immunohistochemical techniques and light (LM) and electron microscopy (EM). Serum HBV DNA was detected in 43.6% of the patients studied. There were no significant differences between HBV DNA positive and DNA-negative patients in terms of their clinical profiles. For HBV DNA-positive patients, the degree of inflammatory cell infiltration and irregular regeneration of hepatocytes was significantly greater than for HBV DNA-negative patients. The cumulative probability of development of hepatocellular carcinoma (HCC) was significantly higher for HBV DNA-positive patients than for HBV DNA negative patients. HBV DNA positivity was a risk factor for the occurrence of HCC according to multivariate analysis. HBsAg and HBcAg were detected in 8.5 and 72.3%, respectively, of the livers of serum HBV DNA-positive individuals. Core particles were detected in the nuclei of the hepatocytes by IEM. The histopathological features and long-term outcome of CH-C or LC could be affected by occult HBV infection. PMID- 19127079 TI - Lipiodol and dye at the site of ablation decreases during RFA. AB - OBJECTIVES: Intrahepatic metastasis is one of the serious complications of radiofrequency ablation (RFA) therapy for hepatocellular carcinoma. The aim of this study was to investigate how liver tissue could be disseminated during the RFA procedure with different devices and protocols in an in vivo porcine model. METHODS: Three pigs underwent RFA procedures using 2 different devices: needles that could be expanded (LeVeen needle) and those that could not (cool-tip needle). A LeVeen needle was used with a single-step full extension method or a stepwise extension method. Before RFA, a mixture of lipiodol and blue dye was injected intrahepatically into a precoagulated area. After the ablation procedure, the specimen was cut to evaluate the amount of dye remaining in the ablated region and the distribution of the dye outside the ablated area. RESULTS: The stepwise extension method resulted in the disappearance of the smallest amount of the dye and lipiodol at the ablation site, compared with the full extension method and cool-tip needle. Dye was found at sites distant from the ablated area in all cases using the cool-tip needle, but in none with the stepwise extension method. CONCLUSIONS: The stepwise procedure using the expandable needle can reduce tumor cell scattering, which can cause intrahepatic metastasis, compared with other methods. PMID- 19127080 TI - Nicotinamide cooperates with retinoic acid and 1,25-dihydroxyvitamin D(3) to regulate cell differentiation and cell cycle arrest of human myeloblastic leukemia cells. AB - Nicotinamide, the amide derivative of vitamin B(3), cooperates with retinoic acid (RA), a form of vitamin A, and 1,25-dihydroxyvitamin D(3) (D3), to regulate cell differentiation and proliferation of human myeloblastic leukemia cells. In human myeloblastic leukemia cells, RA or D3 are known to cause MAPK signaling leading to myeloid or monocytic differentiation and G0 cell cycle arrest. In this process, RA or D3 induces the early expression of CD38, a receptor that causes ERK signaling and propels further differentiation. Our study demonstrates that nicotinamide in combination with RA or D3 affected induced expression levels of CD38, CD11b and CD14, suggesting a cooperative function of nicotinamide and RA or D3. Nicotinamide transiently retarded the initial RA- or D3-induced expression of CD38, which subsequently reached the same nearly 100% expression. Nicotinamide induced ERK activation and further enhanced the RA-induced ERK activation, but the D3-induced ERK activation was diminished by nicotinamide, although levels still exceeded those induced by RA, suggesting lineage-specific nicotinamide responses. Nicotinamide enhanced both RA- and D3-induced CD11b expression, inducible oxidative metabolism, and G0 cell cycle arrest, accelerating their induced occurrence in all instances. Consistent with this, the RA- or D3-induced downregulation of PARP was enhanced by nicotinamide. Nicotinamide thus regulated RA- or D3-induced differentiation and G0 arrest, causing a transient delay in certain early aspects of the progression to terminal differentiation but ultimately accelerating the occurrence of terminally, functionally differentiated G0 cells. PMID- 19127081 TI - Selective killing of leukemia and lymphoma cells ectopically expressing hCGbeta by a conjugate of curcumin with an antibody against hCGbeta subunit. AB - OBJECTIVE: A variety of cancers ectopically express human chorionic gonadotropin beta (hCGbeta). Patients harboring such cancers have poor prognosis and adverse survival. A recombinant chimeric antibody, cPiPP, exhibiting high affinity and specificity for hCGbeta/hCG was engineered. This study was designed to determine whether this antibody alone or conjugated to curcumin can selectively kill tumor cells expressing hCGbeta. EXPERIMENTAL DESIGN: The study was carried out on MOLT 4 and U-937 cells expressing hCGbeta and on peripheral blood leukocytes of acute myeloid leukemia (AML) patients. The anticancerous compound curcumin was conjugated to cPiPP. The binding of cPiPP and cPiPP-curcumin conjugate to cells was studied by flow cytometry and cytotoxicity by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT), FACS with propidium iodide staining, trypan blue exclusion assay and microscopy. RESULTS: The antibody did not impair the growth of MOLT-4 and U-937 cells in culture. Its conjugate with curcumin, however, was lethal to both cell lines. The immunoconjugate killed tumor cells bearing the CD33 marker of an AML patient expressing hCGbeta but did not have a similar action on cells of another AML patient with the CD13 marker but who was negative for hCGbeta. CONCLUSION: A humanized antibody against hCGbeta linked to curcumin has potential for therapy of hCGbeta-expressing tumors. PMID- 19127082 TI - Successful protein C concentrate administration during initiation of oral anticoagulation in adult patients with severe congenital protein C deficiency: report of two cases. AB - Protein C (PC) is a vitamin K-dependent proenzyme with anticoagulant activity, and patients with congenital PC deficiency are at high risk for thrombotic episodes. In patients with PC deficiency, starting treatment with oral anticoagulant drugs is associated with a transient hypercoagulable state and clinically overt thromboembolic complications before reaching a full anticoagulant effect. This report describes a successful supplementation with PC concentrate in two adult patients with moderately severe PC deficiency during the initiation of oral anticoagulation and a course of therapeutic dose of low molecular-weight heparin for acute venous thromboembolism. Plasma PC levels above 50% were observed in both patients and maintained during the entire supplementation treatment period with PC concentrate until a stable therapeutic anticoagulation level has been reached. These results have been obtained within a short time, thus allowing a safe administration of a loading dose of warfarin. No adverse reactions to the PC concentrate, i.e. skin necrosis and other thromboembolic complications, bleedings or allergic reactions, were observed. We conclude that PC concentrate seems to be effective for the prevention of thromboembolic complications and safe in patients with congenital PC deficiency while initiating oral anticoagulants. PMID- 19127083 TI - Major and potential prothrombotic genotypes in patients with venous thrombosis and in healthy subjects from Slovenia. AB - The objective of our study was to investigate the prevalence of the polymorphisms factor V Leiden (FVL), prothrombin G20210A (PT G20210A), methylenetetrahydrofolate reductase C677T (MTHFR C677T), plasminogen activator inhibitor type 1 -675 4G/5G (PAI-1 4G/5G) and factor XII -4 C/T (FXII -4 C/T) in 295 Slovenian patients with venous thrombosis (VT) and 223 healthy controls in order to establish their contribution to the risk for VT. The major genetic risk factor was FVL, while PT G20210A, MTHFR 677 C/T, PAI-1 4G/5G and FXII -4 C/T polymorphisms were not. However, PT G20210A increased the risk of recurrent VT, MTHFR C677T increased the risk in older patients, while the FXII -4 T allele suggested a possible protective effect in younger patients. The risk of VT increased with increasing number of genetic defects. PMID- 19127084 TI - Obesity and activated protein C resistance. AB - It has been reported that obesity may be associated with activated protein C resistance, which could increase the thrombotic risk in these patients. The aim of our study was to evaluate this parameter in obese patients and controls, as well as the effect of weight loss on this parameter. In 63 severely or morbidly obese patients and in 65 healthy volunteers, an anthropometric and analytical evaluation (activated protein C resistance and prothrombin fragment F1 + 2) was performed at baseline and after 3 months of diet. Obese patients showed higher levels of F1 + 2 than controls, whereas activated protein C sensitivity ratios showed no differences. After weight loss, prothrombin fragment F1 + 2 was reduced, but no differences were found in activated protein C sensitivity. We did not find an activated protein C-resistant phenotype in obese subjects. PMID- 19127085 TI - Venous thromboembolism prevention in patients with heart failure: an often neglected issue. AB - Several epidemiological studies have shown a high prevalence of venous thromboembolism (VTE) complications in patients with acute heart failure; in addition, the level of risk associated with this disease is notable, ranging from 15 to 30%. Three large clinical trials have clearly demonstrated the efficacy and safety of pharmacological prophylaxis in internal medicine patients hospitalized for an acute medical disease; on the contrary, until now there are no studies which have evaluated antithrombotic prophylaxis in a selected population of patients with heart failure only. Moreover, discrepancies existing among recommendations reported in different guidelines may produce uncertainties in the management of VTE prevention in patients with heart failure and may contribute to an underuse of thromboprophylaxis in the daily clinical practice. The aim of this review is to analyze the existing evidence about VTE risk in patients with heart failure as well as the efficacy and safety of antithrombotic prevention, and to underline which are the most important unmet clinical issues for the optimal management of thromboprophylaxis in this particular clinical setting. PMID- 19127086 TI - Adenosine diphosphate-induced platelet aggregation correlates with platelet activation identified with the use of flow cytometry. AB - BACKGROUND: Assessment of the likelihood of platelet activation (i.e. platelet reactivity) identifies patients at high and low risk of subsequent thrombotic events. Turbidometric platelet aggregation has been used to assess platelet function for more than 4 decades. We have developed a method to assess individual components of platelet activation with the use of flow cytometry that is performed in minimally altered whole blood. AIMS: To compare assessment of platelet reactivity determined with the use of aggregometry and flow cytometry. MATERIAL AND METHODS: Twenty adult patients with atherosclerotic vascular disease were included in this study. Blood from each patient was used to determine turbidometric platelet aggregation and to assess platelet activation by flow cytometry. ADP was used as the agonist. Values are means +/- SEM. Comparison was performed with the use of Student's t test and correlation was assessed with the use of Pearson's correlation analysis. RESULTS: Both maximal aggregation and the slope of aggregation correlate with the percentage of platelets that bound fibrinogen in response to 0.2 microM ADP. The best correlation was seen between the slope of aggregation induced by 0.2 or 1 microM ADP and the percentage of platelets that bound fibrinogen in response to 0.2 microM ADP (for 0.2 microM r = 0.62, p = 0.038; for 1 microM r = 0.71, p = 0.025). CONCLUSION: The binding of fibrinogen to activated platelets assessed with the use of flow cytometry correlates best with the slope of turbidometric aggregation and appears to reflect the propensity of platelets to activate. PMID- 19127087 TI - Effect of Iron Therapy on Platelet Function among Iron-Deficient Women with Unexplained Menorrhagia. AB - This study was performed to evaluate the effect of iron therapy on platelet function among women with unexplained menorrhagia in order to better understand possible interactions between iron deficiency anemia and platelet behavior and menorrhagia. Platelet aggregation and adenosine triphosphate (ATP) release induced by 5.0 mM adenosine diphosphate (ADP), 0.5 mM arachidonic acid (AA), 1.0 mg/ml ristocetin and 2 microg/ml collagen were studied by whole-blood platelet lumi-aggregometer in 50 menorrhagic women before and after oral iron therapy and in 22 women of the control group. There was a significant increase in AA- induced platelet aggregation (p < 0.05) and a decrease in ristocetin-induced platelet aggregation (p < 0.01) after treatment. Pre- and posttreatment platelet aggregation responses to ADP and collagen were not significantly different (p > 0.05). Pre- and posttreatment platelet secretion responses to all agonists disclosed no significant difference (p > 0.05). There was no significant difference between the study group after treatment and the control group in respect to platelet aggregation and ATP secretion values induced by all agonists (p > 0.05). We conclude that iron deficiency anemia in women causes AA-induced platelet dysfunction, which may give rise to increased menstrual blood loss and can be reversed by iron repletion. PMID- 19127088 TI - Variation in platelet function testing has a major influence on detection of aspirin resistance in healthy subjects. AB - INTRODUCTION: An increased demand for monitoring aspirin treatment by platelet function tests has been observed, but data on the biological variation of these tests are insufficient. The aim of this study was to assess the biological variation of optical platelet aggregometry and closure time in healthy subjects without aspirin and after aspirin ingestion. SUBJECTS AND METHODS: In 20 healthy subjects, blood was sampled 4 times: on 2 consecutive mornings a day after aspirin ingestion (100 mg/daily) and on 2 consecutive days of no treatment. In all samples, arachidonic acid-, ADP- and collagen-induced optical platelet aggregation was measured, and closure times were determined by collagen/epinephrine (CEPI) and collagen/ADP (CADP) cartridges in a platelet function analyzer-100. RESULTS: Aspirin significantly reduced arachidonic acid- and ADP-induced platelet aggregation and significantly prolonged CEPI closure time, but had no significant effect on collagen-induced platelet aggregation and CADP closure time. Aspirin increased both within- and between-subject coefficients of variation. Arachidonic acid-induced platelet aggregation was the most sensitive to aspirin and no aspirin-resistant subjects were detected on either day after aspirin. According to ADP-induced platelet aggregation or CEPI closure time, 25 and 30% of healthy subjects, respectively, changed from aspirin resistant to aspirin responsive or vice versa from one day to another. There was no agreement between platelet function tests in determining aspirin resistance. CONCLUSIONS: A significant variation in optical platelet aggregometry and closure time exists and is presumed to have a major effect on determination of aspirin resistance. PMID- 19127089 TI - Acute versus chronic myocardial ischemia: a differential biological profile study. AB - OBJECTIVE: To determine the possible differences in lipid, thrombogenic and inflammatory marker concentrations and the presence of chronic and acute coronary artery disease (stable and unstable angina, respectively), comparing them with a group of control patients with normal coronary arteries. MATERIAL AND METHODS: This prospective cohort study included 125 patients with unstable angina, 189 with stable angina and a control group of 83 patients with normal coronary arteries. Marker concentrations were measured in all 3 groups. Logistic regression analysis was performed to determine whether such factors could predict unstable or stable angina. RESULTS: Lipid parameter concentrations were similar in the 2 coronary disease groups and significantly lower than in controls. Haemostatic and inflammatory marker concentrations were higher in patients with coronary disease, but were statistically significant only when comparing unstable angina patients with normal controls. Unstable angina patients had significantly higher levels of lipoprotein (a) [Lp(a)], fibrinogen, C-reactive protein (CRP) and leucocytes. Multiple logistic regression analysis showed that CRP (OR 2.635, 95% CI 1.417-4.898), smoking (OR 3.416, 95% CI 1.773-6.584), leucocytes (OR 2.034, 95% CI 1.079-3.836) and Lp(a) (OR 2.269, 95% CI 1.188-4.334) were independent risk factors of unstable versus stable angina. CONCLUSIONS: Patients with unstable angina present a more atherogenic profile than patients with stable angina. Together with smoking, elevated Lp(a), CRP and leucocyte concentrations proved to be associated with the presence of unstable angina. PMID- 19127090 TI - Therapeutic efficacy of leukocytapheresis for procoagulant microparticles during hemophagocytic syndrome. AB - Hemophagocytic syndrome (HPS) presents with signs of persistent remittent fever, hepatosplenomegaly, pancytopenia, hepatic dysfunction, and disseminated intravascular coagulation because of hypercytokinemia caused by activated T lymphocytes and macrophages. In recent years leukocytapheresis using a leukocyte removal filter (known as lymphocytapheresis, LCAP) has been applied to the treatment of various autoimmune diseases. The removal of activated monocytes during LCAP treatment appears useful for hypercytokinemia. We experienced a 32 year-old Japanese man with HPS with elevated tissue factor-enriched monocyte derived microparticles (MDMPs) and pro-inflammatory cytokines/chemokines. Improvements in the level of MDMPs and hypercytokinemia were observed after LCAP treatment. LCAP treatment performed for HPS can be considered a therapeutic strategy for patients with a risk of fetal hemorrhage. PMID- 19127091 TI - Acute complete splenic infarction in a patient with Hirschsprung's disease and literature review of complete splenic infarction. AB - Here we report a case of Hirschsprung's disease presenting with acute complete splenic infarction due to thrombus in the splenic vena. MTHFR C677T (methylenetetrahydrofolate) gene homozygote mutation was a risk factor for thrombosis. According to our knowledge, this is the first report for a Hirschsprung's disease patient with acute complete splenic infarct due to isolated splenic vein thrombosis accompanied by MTHFR C677T gene homozygote mutation. PMID- 19127092 TI - Upper-extremity deep vein thrombosis in a patient on clozapine therapy carrying the prothrombin G20210A mutation. AB - Clozapine treatment for resistant schizophrenic disorders has been associated to venous thromboembolic events. We report the case of a patient who developed upper extremity deep vein thrombosis just 2 months after starting on clozapine in whom the thrombophilia work-up revealed the presence of the prothrombin G20210A mutation. PMID- 19127093 TI - Transient ventriculomegaly in an adolescent presenting with shunted hydrocephalus, diabetic ketoacidosis, and hyperglycemia. AB - In this report, the authors describe a unique presentation of ventriculomegaly in the setting of diabetic ketoacidosis (DKA). A 15-year-old male, with a history of shunt placement for hydrocephalus and repair of a myelomeningocele, presented to the emergency room with DKA and was found to have ventriculomegaly. At the time of presentation, the patient had a 24- to 48-hour history of lethargy, nausea, vomiting, fever (102.2 degrees F), and polydipsia. A computed tomographic (CT) scan of the head indicated increased ventricular size compared with previous studies, thus prompting a neurosurgical evaluation. Blood glucose level at admission was found to be 1,551 mg/dl, bicarbonate level was 9 mmol/l, and pH was 7.08. The patient was treated for DKA; his fever, lethargy, nausea, and vomiting subsequently resolved. A repeat CT scan obtained 24 h after admission revealed a return of the ventricular system to its baseline size. On the basis of radiographic and laboratory evaluations, the authors hypothesize that the transient ventriculomegaly was directly related to extreme hyperglycemia. PMID- 19127094 TI - A novel TP53 germline mutation in a family with a history of multiple malignancies: case report and review of the literature. AB - OBJECTIVE: Choroid plexus carcinoma (CPC) has been associated with TP53 germline mutations and Li-Fraumeni syndrome (LFS). We describe our finding of a novel germline mutation in the TP53 gene in a family with multiple malignancies and in association with a child presenting with CPC. METHOD: An 8-month-old male presented with seizure-like activity; imaging disclosed a 1.5-cm left ventricular mass confirmed to be CPC intra- and postoperatively. Family history was significant for a half-sister who died of a primary CNS sarcoma and a paternal grandmother negative for BRCA1, BRCA2, MLH1, and MSH2 mutations with multiple (>6) LFS spectrum malignancies. RESULTS: Familial TP53 testing revealed an A-->T substitution at DNA position 13071, creating a deleterious Asn-->Ile substitution at amino acid 131 in exon 5. CONCLUSION: Physicians treating patients with CPC should be attuned to reviewing family history for risk factors suggestive of genetic cancer syndromes such as LFS. These syndromes markedly influence both the patient and family members and may alter postoperative treatment regimens. PMID- 19127095 TI - Holocord hydrosyringomyelia with terminal myelocystocele revealed by constructive interference in steady-state MR imaging. AB - We report an operated case with terminal myelocystocele and holocord hydrosyringomyelia (syrinx). The patient exhibited a subcutaneous mass at the right lumbosacral region with multiple anomalies including scoliosis and hyperdactylia of the right foot and systemic disease such as hydronephroureter. Neurological examination revealed palsy of the left diaphragm, and left upper and right lower limbs. Constructive interference in steady-state (CISS) images demonstrated a terminal myelocystocele with a huge syrinx extending to the medulla. The curvilinear reconstruction of the CISS sequence depicted continuous cystocele and whole syrinx. At day 37, sac excision and untethering of the spinal cord were performed. The caudal part of the central canal was opened to the subarachnoid space (terminal ventriculostomy), resulting in a slight improvement of limb palsy and shrinkage of the holocord syrinx. CISS imaging is useful to detect the complicated pathology, and terminal ventriculostomy should be performed to improve associated syrinx. PMID- 19127096 TI - Unique defect representing features of Chiari type III and IV malformations. PMID- 19127097 TI - Giant occipital encephalocele associated with microcephaly and micrognathia. PMID- 19127098 TI - A witnessed short fall mimicking presumed shaken baby syndrome (inflicted childhood neurotrauma). PMID- 19127099 TI - Presumed shaken baby syndrome. PMID- 19127100 TI - Neuropsychological functioning in schizophrenia patients with first-rank (passivity) symptoms. AB - BACKGROUND: In this study, we conducted a broad investigation of neuropsychological functioning in schizophrenia patients with first-rank (passivity) symptoms (FRS) using standardized cognitive tests. We focused on 3 neuropsychological domains, partly informed by 3 broad hypotheses concerning the pathogenesis of FRS. We tested predictions that patients with FRS would show: (i) memory deficits consistent with abnormalities in medial temporal lobe functioning; (ii) reduced cerebral lateralization on tasks of motor and language functions, and (iii) deficits on tasks of executive functions as a measure of the integrity of the prefrontal cortex for supporting self-monitoring processes. SAMPLING AND METHODS: In 2 parallel studies, we administered a range of neuropsychological tests to patients with FRS (sample A = 17, sample B = 15) and without FRS (sample A = 28, sample B = 20) and to healthy controls (sample A = 109, sample B = 22). RESULTS: The results showed reduced cerebral lateralization in patients with FRS, but there were no memory deficits or executive dysfunctions relative to patients without FRS. An unexpected finding was that, on many cognitive tasks, the performance of the patients with FRS was relatively spared compared to that of the patients without FRS. These results could not be accounted for by demographic features or medication effects. CONCLUSIONS: Our results provide qualified support for the reduced lateralization hypothesis. Analyses of performance across neuropsychological domains showed that generalized deficit is not a sufficient explanation for the occurrence of FRS. PMID- 19127101 TI - Effects of symptom reduction and psychotropic medication on cognitive impairment in depression. AB - BACKGROUND: Depression is related to cognitive performance. This follow-up study examines the influence of depression symptoms and psychopharmacological treatment on change in the cognitive performance of patients with depressive episodes over a 2-year period. SAMPLING AND METHODS: Sixty-two in- and outpatients with depression of varying severity (ICD-10: F31-F33) were examined in an open prospective naturalistic observational study with 3 points of measurement and tested by use of 3 computerized cognitive performance tests [Visual Attentiveness Test (VAT), Continuous Attention Test (CAT), Word Recognition Test (WRT)], while the psychotropic medication was classified by subclass and dosage. Statistical analysis was performed by random-effects regression models. RESULTS: The raw values of VAT speed, CAT speed and WRT quality improved over time. However, the positive time trend disappeared after the patients' clinical and personal characteristics were controlled for. The processing speed of the VAT was found to be negatively influenced by depressive symptoms. That of the CAT developed favorably with increasing level of education. The performance qualities of the VAT, WRT and CAT were positively related to the participants' educational level. The patients who received antipsychotic treatment performed worse on WRT quality than those who were not treated with antipsychotics. CONCLUSIONS: The cognitive performance was relatively stable during the treatment process and not affected by clinical characteristics or type of medication. Cognitive deficits in patients with depression could be a trait rather than a state marker. PMID- 19127102 TI - Mood and fatigue in everyday life are temporally related to cellular immune activity. Preliminary evidence from an extensive observation of a patient with prior breast cancer. PMID- 19127103 TI - Extended-spectrum beta-lactamases: implications for the clinical laboratory and therapy. AB - Production of extended-spectrum beta-lactamase (ESBL) is one of the most important resistance mechanisms that hamper the antimicrobial treatment of infections caused by Enterobacteriaceae. ESBLs are classified into several groups according to their amino-acid sequence homology. While TEM and SHV enzymes were the most common ESBLs in the 1990s, CTX-M enzymes have spread rapidly among Enterobacteriaceae in the past decade. In addition, some epidemiological studies showed that organisms producing CTX-M enzymes had become increasingly prevalent in the community setting in certain areas in the world. Several novel enzymes with hydrolyzing activity against oxyimino-cephalosporins, albeit with additional enzymatic characteristics different from those of original TEM and SHV ESBLs (e.g., inhibitor-resistance), have been discovered and pose a problem on the definition of ESBLs. Although several methods to detect the production of ESBL are available in clinical laboratories, existence of other factors contributing resistance against beta-lactams, e.g., inducible production of Amp-C beta lactamase by some species of Enterobacteriaceae, or inhibitor-resistance in some ESBLs may hinder the detection of ESBLs with these methods. Carbapenems are stable against hydrolyzing activity of ESBLs and are regarded as the drug of choice for the treatment of infections caused by ESBL-producing Enterobacteriaceae. Although several other antimicrobial agents, such as fluoroquinolones and cephamycins, may have some role in the treatment of mild infections due to those organisms, clinical data that warrant the use of antimicrobial agents other than carbapenems in the treatment of serious infections due to those organisms are scarce for now. PMID- 19127104 TI - [Correlation of chromosomal aberrations with prognostic markers in multiple myeloma patients--a single institution study]. AB - BACKGROUND: Immunoglobulin heavy chain (IGH) gene rearrangement, 13q14 deletion and trisomy 1q are frequently observed in Korean patients with multiple myeloma. The purpose of our study was to analyze the statistical correlation between chromosomal aberrations and routine laboratory test results as prognostic markers and to evaluate the potential of chromosomal aberrations for the indirect assessment of prognosis in multiple myeloma patients. METHODS: We investigated the prevalence of cytogenetic aberrations in 41 patients with newly diagnosed multiple myeloma. Cytogenetic analysis was conducted by conventional karyotyping and FISH for the presence of IGH/CCND1 translocation, 13q14 deletion, and trisomy 1q using bone marrow aspirates. The records of routine laboratory tests were reviewed and their correlation with cytogenetic abnormalities was investigated. RESULTS: Sixteen (39.0%) of 41 patients had at least one cytogenetic abnormalities in conventional karyotyping or FISH. In FISH analysis of 37 patients, 8 (21.6%) showed positive result for IGH/CCND1 translocation, 8 (21.6%) for trisomy 1q, and 5 (13.5%) for 13q14 deletion. Cytogenetic abnormalities, especially trisomy 1q, were associated with significantly lower hemoglobin level and significantly higher bone marrow plasma cell percentage and beta(2) microglobulin level. CONCLUSIONS: Statistical correlation between the presence of trisomy 1q and prognostic markers suggests that the evaluation of trisomy 1q in multiple myeloma patients may be used for the indirect assessment of prognosis in these patients. PMID- 19127105 TI - [Utility of D-dimer assay for diagnosing pulmonary embolism: single institute study]. AB - BACKGROUND: Pulmonary embolism (PE) presents with diverse non-specific signs and symptoms and its diagnosis mainly depends on diagnostic imaging tests which are laborious and not cost-effective, and only a small proportion of patients with suspected PE actually have the disease. The aim of this study was to analyze the utility of D-dimer test for diagnosing PE by categorizing patients into 'PE likely' and 'PE unlikely' groups using Wells score for clinical probability. METHODS: One hundred forty consecutive patients with clinically suspected PE, in whom D-dimer and imaging tests were performed were enrolled. Dignosis of PE was made when the imaging tests were positive. Wells scores were retrospectively assigned and the dignostic utility of D-dimer test was analyzed. RESULTS: Of the 140 patients studied, D-dimer test was positive in 97 and diagnostic imaging tests revealed PE, deep vein thrombosis (DVT), and PE+DVT in 24, 3, and 7 patients, respectively. For the diagnosis of PE, D-dimer test with cutoff value of > or =230 ng/mL showed sensitivity, specificity, and negative predictive value of 96.8%, 39.6%, and 97.7%, respectively. These values were 96.3%, 37.9%, and 91.7% in 'PE likely' group (n=56), and 100%, 38.8%, and 100% in 'PE unlikely' group (n=84). Among 43 patients with D-dimer values of <230 ng/mL, only one patient was diagnosed with PE, who belonged to the 'PE likely' group. CONCLUSIONS: D-dimer test cannot be used as a stand-alone test to diagnose PE, but it can be helpful for exclusion of PE especially in 'PE unlikely' group according to Wells score. PMID- 19127106 TI - [Four cases of hematologic malignancy following radioactive iodine therapy for thyroid cancer]. AB - Ionizing radiation including I(131) might produce chromosomal translocation, causing hematologic malignancy. The incidence of leukemia following radioactive iodine treatment for thyroid cancer has been reported to be approximately 0.1 to 2.0% in Western countries, whereas fewer cases have been reported in Korea. We hereby report four cases of secondary hematologic malignancy, who received iodine therapy for thyroid cancer after thyroidectomy: two cases of acute lymphoblastic leukemia with t(9;22)(q34;q11.2), a case of MDS with 5q deletion, and a case of MDS with normal karyotype. Three cases of hematologic malignancy have developed after cumulative dosage of less than 800 mCi. The treatment intervals in two cases were less than 12 months, and the other two cases had I(131) therapy only once. Assessment of causality using the Naranjo probability scale for adverse drug reactions showed that a 'possible' relationship existed between the use of I(131) and secondary hematologic malignancy in all of the four cases in this report. PMID- 19127107 TI - Human telomerase reverse transcriptase (hTERT): a target molecule for the treatment of cisplatin-resistant tumors. AB - BACKGROUND: Human telomerase reverse transcriptase (hTERT) is a catalytic enzyme that is required for telomerase activity (TA) and cancer progression. Telomerase inhibition or inactivation increases cellular sensitivity to UV irradiation, DNA damaging agents, the tyrosine kinase inhibitor, imatinib, and pharmacological inhibitors, such as BIBR1532. hTERT is associated with apoptosis. Some patients show drug-resistance during anti-cancer drug treatment and the cancer cell acquire anti-apoptotic mechanism. Therefore, we attempted to study correlation between hTERT and drug-resistance. METHODS: To study the correlation between protein level and activity of hTERT and drug-resistance, Western blotting and telomerase repeat amplification protocol (TRAP) assays were performed. To investigate whether hTERT contributes to drug resistance in tumor cells, we transiently decreased hTERT levels using small interfering RNA (siRNA) in T24/R2 cells. RESULTS: hTERT knockdown increased Bax translocation into the mitochondria and cytochrome C release into the cytosol. Caspase inhibitors, especially Z-VAD FMK, rescued this phenomenon, suggesting that the stability or expression of hTERT might be regulated by caspase activity. CONCLUSIONS: These data suggest that hTERT might be a target molecule for drug-resistant tumor therapy. PMID- 19127108 TI - [Measurement of amylase in saliva collected by salivette]. AB - BACKGROUND: Saliva is increasingly being used as a specimen for systemic disease as well as for oral health status. Especially, salivary amylase has been studied as an excellent index for psychological stress. Authors evaluated the measurement of salivary amylase activities collected by Salivettes (Sarstedt, Germany). METHODS: Saliva specimens were collected from 13 healthy adults between 10:00 and 11:00 a.m. Participants were asked to gently chew tampons of Salivettes for 1 min. Immediately after collection, all specimens were stored frozen. On the day of testing, they were centrifuged after thawing and diluted with distilled water. Amylase was measured by Dimension RxL Max (Dade Behring Inc., USA). We evaluated precision, linearity, and recovery rate of Salivette. Amylase activities between collection of saliva by Salivette and passive drool were compared, and also the changes of amylase by the storage temperature were evaluated. RESULTS: Intra-run CVs for three levels of amylase were excellent. Between-day CVs and total CVs were good only for mid and high levels. A good linear relationship was found at all diluted levels. Dosing Salivettes with 2 mL, 1.5 mL, and 1 mL yielded sample recovery 85.5+/-2.4%, 82.4+/-1.5%, and 72.2+/-3.1%, respectively and amylase recovery 78.9+/-10.9%, 74.1+/-13.7%, and 37.3+/-26.9%, respectively. Amylase by Salivette and passive drool were correlated well (r=0.757), although they showed a significant difference. Amylase activity was not affected by the storage temperature. CONCLUSIONS: Measurement of salivary amylase using Salivette could be a useful test having good intra-run CVs and linearity. More than 1.5 mL of saliva would be needed to have more than 70% recovery of Salivette. PMID- 19127109 TI - [Two cases of vancomycin-intermediate Staphylococcus aureus isolated from joint tissue or wound]. AB - Since its first isolation in 1997, vancomycin-intermediate Staphylococcus aureus (VISA) has been a clinical concern because it may lead to treatment failure. Up to the present, there were two reports of clinical VISA cases in Korea. We now report two additional cases of VISA with the minimum inhibitory concentration (MIC) of 4 microg/mL. The first patient was a 59 yr-old man who had undergone total hip replacement arthroplasty in 1999 due to avascular necrosis of femur heads. He had recurrent episodes of infected hip caused by methicillin-resistant Staphylococcus aureus (MRSA) and was treated with vancomycin. He underwent replacement operation of prosthesis. Cultures of joint fluid and joint tissue grew S. aureus. Vancomycin MIC as determined by a broth microdilution method was 4 microg/mL for the both isolates. The patient was treated with high enough doses of vancomycin to maintain serum trough concentrations at 20-25 microg/mL for 52 days and was discharged. The second patient was a 57 yr-old man with diabetes. He lost consciousness from drinking. After recovery of consciousness, he was diagnosed with aspiration pneumonia. MRSA and Acinetobacter baumannii were cultured from sputum and the patient was treated with vancomycin and meropenem. During hospitalization, bed sores developed in his ankle and back. A wound culture from the sore grew S. aureus with vancomycin MIC of 4 microg/mL. Since infection was localized, systemic antibiotics did not seem necessary, and the patient was transferred to another hospital for isolation and management. PMID- 19127110 TI - [Production and characterization of anti-staphylococcal toxic shock syndrome toxin-1 monoclonal antibody]. AB - BACKGROUND: Recently the association between the virulence factors of Staphylococcus aureus and the outcome of the patients infected with the organism appears to be the subject of active investigation. Toxic shock syndrome toxin-1 (TSST-1) is thought to be a clinically more significant virulence factor than other staphylococcal toxins. We attempted to produce and characterize monoclonal antibodies to staphylococcal TSST-1. METHODS: An important epitope of TSST-1, amino acids 1-15 region, was synthesized into a peptide antigen, and Balb/c mice were immunized by intraperitoneal injection of the synthetic antigen. Hybridomas were produced by fusing immunized murine splenocytes with immortal myeloma cells. Hybridomas were cloned through a limiting dilution method. Stable cultured hybridoma was injected into the peritoneal cavity of Balb/c mice, and peritoneal fluid containing the monoclonal antibody was produced. RESULTS: One IgG(2b) type monoclonal antibody and two IgM type monoclonal antibodies were obtained. The IgG(2b) type monoclonal antibody was able to detect 5 microg of TSST-1 with Western blot analysis and showed a strong reactivity to TSST-1 with ELISA. CONCLUSIONS: Highly immunoreactive anti-TSST-1 monoclonal antibody was produced by the use of synthesized peptide antigen. Diagnostic and protective capacity of this monoclonal antibody should be evaluated in the future. PMID- 19127111 TI - [Diagnostic utility of AxSYM anti-cyclic Citrullinated peptide antibody assay]. AB - BACKGROUND: The presence of rheumatoid factor (RF) is one of the classification criteria of the American College of Rheumatology (ACR) for rheumatoid arthritis (RA), but it has a limitation of low specificity. We compared the diagnostic utility of anti-cyclic citrullinated peptide (CCP) antibodies analyzed by an automated immunoassay system with that measured by a 96 well plate ELISA method. METHODS: The RF and anti-CCP antibodies were determined in 172 serum samples: 52 RA patients, 73 disease controls (systemic lupus, Sjogren's syndrome, palindromic rheumatism), and 47 healthy controls. Anti-CCP antibodies were measured by DIASTAT 96 well plate ELISA method (Axis-Shield Diagnostics, UK) and AxSYM automated microparticle enzyme immunoassay system (Abbott Laboratories, USA). RF was assayed by latex immunoturbidimetry (Toshiba 200 FR, Japan). The diagnostic performances of these tests were compared using a ROC curve analysis, and linearity and precision analysis of AxSYM anti-CCP was carried out. RESULTS: The sensitivities of RF, DIASTAT anti-CCP, and AxSYM anti-CCP were 78.8%, 84.6%, and 82.7%, respectively and the specificities were 72.5%, 88.3%, and 88.3%, respectively. On ROC curve analysis, the area under the curve was 0.924 for AxSYM anti-CCP, 0.886 for DIASTAT anti-CCP, and 0.847 for RF. AxSYM anti-CCP showed a good linearity, and within-run and total-run precision. CONCLUSIONS: Diagnostic performance of automated AxSYM anti-CCP assay was comparable to that of DIASTAT 96 well plate ELISA method. AxSYM anti-CCP assay has an advantage of random access capability and will be useful in laboratories with low sample number and/or with a need of rapid turnaround time. PMID- 19127112 TI - [Allele and haplotype frequencies of human leukocyte antigen-A, -B, and -DR loci in Koreans: DNA typing of 1,500 cord blood units]. AB - BACKGROUND: The HLA system is known to be the most polymorphic genetic system in human, and HLA allele and haplotype distribution varies widely among different ethnic groups. This study was performed to examine the frequencies of HLA alleles and haplotypes in Koreans. METHODS: We examined HLA-A, -B, and -DR alleles at the serologic level in 1,500 cord blood units obtained from Koreans using the PCR sequence specific oligonucleotide (SSO) method. Allele and haplotype frequencies were estimated by the maximum likelihood method using the computer program developed for the 11th International Histocompatibility Workshop. RESULTS: HLA alleles found in Koreans were 12 in A, 31 in B, and 13 in DR loci. Most frequent alleles with frequencies > or =10% in each locus in decreasing order of frequency were: A2, A24, A33, A11; B62; DR4, DR15, DR9, and DR13. Two-locus haplotypes with frequencies > or =0.1% were 104 A-B and 115 B-DR haplotypes, among which those with frequencies > or =1.0% showing significant positive linkage disequilibrium (P< or =0.001) were 21 A-B and 20 B-DR haplotypes. A total of 169 A-B-DR haplotypes with frequencies > or =0.1% were identified. The results were similar to those of a previous study in 1,600 Koreans, although some differences were noted in the distribution of some less frequent alleles or haplotypes with frequencies < or =0.5%. CONCLUSIONS: We provided the allele and haplotype frequencies of HLA-A, -B, and -DR in cord blood units of Korean ethnicity defined by a DNA typing method, which can be used as basic data on Koreans for organ transplantation and disease association studies. PMID- 19127113 TI - [Evaluation of automated architect syphilis TP as a diagnostic laboratory screening test for syphilis]. AB - BACKGROUND: The aim of the study was to establish a new syphilis test algorithm using Architect Syphilis TP (Abbott Japan, Japan: AST), a fully automated treponemal antibody test, as a screening test in a university hospital laboratory. We evaluated performance characteristics of AST in various patient groups. METHODS: A total of 1,357 serum samples obtained from patients at a university hospital from June to August, 2008 were categorized into checkup, preoperative, other diseases, diagnosis (clinically suspected of syphilis), and follow up groups. We compared the results of AST with those of RPR (N=1,276) or Treponema pallidum hemagglutination assay (TPHA, N=81). Samples with discrepant results between RPR or TPHA and AST were retested by fluorescent treponemal antibody absorption test (FTA-ABS) and all patients' clinical records were thoroughly reviewed. RESULTS: The positive rate of AST was significantly higher than that of RPR in preoperative and other diseases groups and was the same as that of RPR in diagnosis group. There were no significant differences in check up and follow up groups. The results of AST showed 97.4% (1,243/1,276) and 97.5% (79/81) concordance rates with those of RPR and TPHA, respectively. Among 26 RPR AST discrepant and FTA-ABS confirmed cases, there were 20 RPR false-negatives, 4 RPR false-positives, 1 AST false-negative, and 1 AST false-positive. CONCLUSIONS: Based on the results and literature review, we established a new syphilis test algorithm using AST as a screening test, which would be helpful for detection of more syphilis patients including latent infections. PMID- 19127114 TI - Molecular and clinical characteristics of myotonic dystrophy type 1 in koreans. AB - BACKGROUND: Myotonic dystrophy type 1 (DM1) is an autosomal-dominant muscular dystrophy caused by expansion of cytosine-thymine-guanine (CTG) trinucleotide repeats in the myotonic dystrophy protein kinase (DMPK) gene. The clinical features of DM1 are multisystemic and highly variable, and the unstable nature of CTG expansion causes wide genotypic and phenotypic presentations. The aim of this study was to characterize the molecular and clinical spectra of DM1 in Koreans. METHODS: The CTG repeats of 283 Korean individuals were tested by PCR fragment analysis and Southern blot. The following characteristics were assessed retrospectively: spectrum of CTG expansions, clinical findings, genotype phenotype correlation, anticipation, and genetic instability. RESULTS: One hundred twenty-four patients were confirmed as DM1 by molecular tests, and the CTG expansions ranged from 50 to 2,770 repeats (median 480 repeats). The most frequent clinical features were myotonia, muscular weakness, and family history. Patients with muscular weakness or dysfunction of the central nervous system harbored larger CTG expansions than those without each symptom (P<0.05). The age of onset was inversely correlated with the size of the CTG expansion (gamma= 0.422, P<0.001). The instability of CTG expansion representing as the maximum difference between sibships was observed from 50 to 700 repeats in nine families. Clinical anticipation and the increase in CTG repeat were significantly higher in maternally transmitted alleles (P=0.002). CONCLUSIONS: Molecular genetic tests are not only essential for diagnosis, but also helpful for suggesting the spectrum and relationship between genotype and phenotype in Korean DM1 patients. PMID- 19127115 TI - Genetic counseling can influence the course of a suspected familial cancer syndrome patient: from a case of Li-Fraumeni like syndrome with a germline mutation in the TP53 gene. AB - We report a 26-yr-old female patient with bilateral breast cancer who was clinically diagnosed with Li-Fraumeni like syndrome (LFL) and subsequently found to have a germline mutation of the TP53 gene. The patient was initially diagnosed with right breast cancer at age 24 yr and then with left breast cancer at age 25 yr. Surgery and radiotherapy were performed accordingly. The patient had a family history of various types of early onset cancers and was referred to a genetic counseling clinic. She was clinically diagnosed with LFL. Genetic analysis of the TP53 tumor suppressor gene was performed with the patient's consent. Direct sequencing of TP53 gene exons 5, 6, 8, 9, and 11 revealed a ermline missense mutation, resulting in an amino acid change from an arginine to a histidine (g.13203G>A, p.R175H). Considering the family history, individualized cancer surveillance was performed including a gastroscopy and a brain MRI. Even though the patient had not shown any neurological symptoms, a huge mass on the temporal lobe was incidentally found and the patient received surgery and radiotherapy. Although the residual mass required further treatment, the patient decided on supportive care alone and was discharged. We report a case of LFL, with a germline TP53 mutation, which was confirmed by gene sequencing in Korea. This case shows how genetic predisposition screening and counseling in patients, suspected of having a familial cancer syndrome, can influence the course of the patient. PMID- 19127116 TI - PI3K: A rational target for ovarian cancer therapy? PMID- 19127117 TI - Potentiation of oncolytic virotherapy with armed shRNA. PMID- 19127118 TI - Epigenetic downregulation of the DNA repair gene MED1/MBD4 in colorectal and ovarian cancer. AB - MED1 is a base excision repair enzyme that interacts with the mismatch repair protein MLH1 and maintains genomic integrity by binding methylated DNA and repairing spontaneous deamination events. MED1 mutations have been associated with microsatellite instability and accelerated colorectal cancer (CRC) tumorigenesis. We propose that promoter methylation may serve as an alternative epigenetic mechanism for MED1 gene suppression during sporadic CRC tumorigenesis. Methylation status of the MED1 promoter was investigated in a panel of ovarian and colorectal cancer cell lines. The MED1 promoter region was sequenced following bisulfite treatment and sequence analysis identified a CpG island within the MED1 promoter which is frequently and preferentially methylated (> or =50%) in ovarian and colorectal cancer cell lines with low/reduced MED1 expression. In vitro reversal of methylation restored MED1 expression. In colorectal cancer patients, when MED1 methylation was present, both tumor and matched mucosa were affected equally (mean frequency of methylation 24%) and there was no correlation between methylation and tumor stage. Patients without history of CRC showed significantly lower frequency of methylation (mean 14%, p < 0.05). Decreased MED1 transcript levels were observed in matched normal mucosa when compared to controls (median fold difference 8.0). Additional decreased expression was seen between mucosa and matched tumor (median fold decrease 4.4). Thus, MED1 promoter methylation and gene silencing occur in sporadic CRC patients and represent an early event in CRC tumorigenesis. Detection of MED1 methylation and gene suppression in normal colon mucosa may contribute to identifying patients at higher risk of developing CRC during screening procedures. PMID- 19127119 TI - Erythropoeisis-stimulating agents (ESAs) in cervix cancer: The "black box" paradox. PMID- 19127120 TI - Ceramide-mediated apoptosis following ionizing radiation in human prostate cancer cells: PKCalpha joins the fray. PMID- 19127121 TI - Combination therapy with TRAIL: Recent developments and potential pitfalls. PMID- 19127123 TI - Current and emerging treatments for mild/moderate acute ambulatory pain. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs), the mainstay of therapy for acute musculoskeletal pain, are among the most widely used drugs in the world. The major limitation of traditional (nonselective) NSAIDs is gastrointestinal (GI) toxicity, which includes dyspepsia, mucosal injury, and serious events, such as ulcers, GI bleeding, and perforation. This article reviews the mechanism of action, safety, and efficacy of traditional and selective NSAIDs (coxibs); risk factors for GI complications; and strategies for reducing risk and preventing NSAID-related gastropathy and cardiovascular complications. The article also discusses the use of aspirin for cardioprotection in patients taking analgesic/anti-inflammatory agents for musculoskeletal pain and the need to balance competing GI and cardiovascular risks. Topical NSAIDs and other formulations that may prove to be safer than currently available agents also are addressed. PMID- 19127124 TI - The pharmacist's role in the nonsteroidal anti-inflammatory drug selection process. AB - The pharmacist provides an integral role in pain management and treatment by focusing on the selection and evaluation of analgesic agents in a process that is patient specific and patient centered. Counseling patients (and the families of patients) who are using nonsteroidal anti-inflammatory drugs (NSAIDs) for acute musculoskeletal pain and inflammation regarding the appropriate use of these agents is a key component of the pharmacist's overall pharmacotherapeutic role. This article reviews the importance of explaining the therapeutic and nontherapeutic effects of NSAIDs and cautions, contraindications, dosing parameters, and the avoidance of multiple NSAID use to patients and prescribers. The article also discusses the need to evaluate the cytochrome P450 system and the patient's pharmacotherapy and comorbid disease history to identify potential drug interactions. Evaluation of patients for comorbidities, allergies, and gastrointestinal, renal, hepatic, hematologic, and cardiovascular risks is also addressed, as are essential laboratory tests and the special needs of elderly patients. PMID- 19127125 TI - The payer side: patient outcomes and cost. AB - Payers and clinicians share a commitment to successful clinical outcomes in patients with acute or chronic pain. However, there are several issues that concern payers and managed care plans, including the cost of therapy and the cost of treating side effects. This article reviews the economic implications of pain management and the need to provide care in a clinically and economically responsible way. The article also addresses the value of medications to patients, the need for more judicious use of very expensive medications, and the use of evidence-based treatment guidelines. PMID- 19127129 TI - Treatment of mild-to-moderate acute pain: what can we do to improve the standard of care? PMID- 19127126 TI - Treatment of Mild-to-Moderate Acute Pain. Panel discussion. PMID- 19127130 TI - Current management of acute musculoskeletal pain in the ambulatory care setting. AB - Three issues related to the management of acute musculoskeletal pain in the ambulatory care setting deserve more attention than they currently receive: pain assessment, the degree of diagnostic specificity needed to select treatment, and gaps in the care that clinicians need to consider. This article describes several pain assessment instruments and explains why they are appropriate in the acute care setting. It also reviews several reports demonstrating that most patients with acute musculoskeletal pain, particularly low back pain, cannot be given a definitive pathoanatomical diagnosis. Therefore, categorizing the pain as mechanical or secondary to underlying causes is sufficient to select and institute treatment. Gaps in care, such as the need for increased patient education, accurate information to dispel misconceptions about therapy, and additional safety and efficacy data about drugs used to treat acute musculoskeletal pain, are also addressed. PMID- 19127131 TI - The drug eluting stent challenge. PMID- 19127132 TI - Pioglitazone and the risk of myocardial infarction and other major adverse cardiac events: a meta-analysis of randomized, controlled trials. AB - A recent meta-analysis suggested that the use of rosiglitazone increases the risk of myocardial infarction (MI) in patients with type 2 diabetes mellitus. It is unclear whether this is a class effect of thiazolidinediones (TZD). We did a meta analysis to evaluate cardiovascular outcomes with the use of pioglitazone. Randomized, controlled trials in which pioglitazone was compared with placebo or other hypoglycemic agents were considered for analysis. Studies were included if the data for MI were available. Studies were identified with use of relevant search words in Medline, Pubmed, EMBASE, CINAHL, and Cochrane databases. Data abstraction was done by 2 individual authors using a standardized protocol. The relative risk across all study groups was computed by the Mantel-Haenszel method, and interstudy heterogeneity was assessed by the chi method. All results were computed according to 95% confidence intervals. Five trials (N = 9965) met the inclusion criteria for analysis. The relative risk for MI was 0.86 (0.69-1.07; P = 0.17). The relative risks for stroke and revascularization were 0.79 (0.61 1.02; P = 0.07) and 0.40 (0.13-1.23; P = 0.11), respectively. Pioglitazone does not increase the risk for MI and may decrease the risk for stroke and revascularization. PMID- 19127133 TI - Cost-effectiveness of dalteparin versus unfractionated heparin as venous thromboembolism prophylaxis in malignant gynecologic surgery. AB - Venous thromboembolic events (VTEs) are serious complications that may occur in the patient undergoing surgery for gynecologic malignancies. The American College of Chest Physicians recommends unfractionated heparin or low-molecular weight heparin as prophylaxis for deep vein thrombosis and pulmonary embolism in this patient population. Cost-effectiveness analyses comparing unfractionated heparin 3 times a day versus once daily dalteparin using published efficacy and safety data demonstrate cost savings if dalteparin were routinely utilized as VTE prophylaxis. Sensitivity analyses support this finding at the upper end of the range of reported proximal DVT, nonfatal pulmonary embolism, and major bleeding incidences. These findings should be viewed as preliminary, and institutions are encouraged to perform their own cost-effectiveness studies in this patient population. PMID- 19127134 TI - An evaluation of the outside therapy of diabetic ketoacidosis in pediatric patients. AB - BACKGROUND: Despite literature outlining suggested initial therapy for pediatric patients with diabetic ketoacidosis (DKA), our impression has been that there may be variations from these recommendations during the initial therapy of pediatric patients with DKA. In order to improve education initiatives, an understanding of the deviations from current practice is required. METHODS: Patients admitted to the pediatric intensive care unit with a diagnosis of DKA were identified from the admission log. The pre-pediatric intensive care unit care including laboratory evaluation, insulin dosing, and fluid therapy was recorded. RESULTS: The study cohort included 135 episodes of DKA in 127 patients (age range: 10 months to 21 years). A complete blood count was obtained in 83.7% of the patients. Serum electrolytes, blood urea nitrogen, and creatinine were obtained in 89.6%, and a serum pH was obtained in 58%. Seventy-two patients received a bolus dose of insulin. The insulin bolus was < or =0.05 units/kg in 1 patient, >0.05 to < or =0.1 units/kg in 13 patients, >0.1 to < or =0.2 units/kg in 27 patients, and >0.2 units/kg in 31 patients. The route of administration for the insulin bolus was intravenous (IV) in 58 patients, a combination of IV and subcutaneous in 7 patients, subcutaneous in 6, and a combination of intramuscular and IV in 1 patient. Fluid administered before transport ranged from 0 to 60.6 mL/kg. Sixteen patients did not receive a fluid bolus. Normal saline was used in 115 patients, Ringer's lactate solution in 3, and 5% glucose in (1/2) normal saline in 1. Seventeen patients (12.6%) received IV sodium bicarbonate. CONCLUSIONS: Major issues with the prehospital care of children and adolescents with DKA included lack of appropriate laboratory evaluation, excessive insulin dosing (both bolus doses and infusion rates), lack of fluid resuscitation, use of inappropriate fluids for resuscitation, and the use of sodium bicarbonate. PMID- 19127135 TI - Preliminary experience with dexmedetomidine for monitored anesthesia care during ENT surgical procedures. AB - Dexmedetomidine is an alpha2-adrenergic agonist that produces anxiolysis, amnesia, sedation, potentiation of opioid analgesia, and sympatholysis. It is currently approved by the U.S. Food & Drug Administration for the sedation of adults in the intensive care setting for up to 24 hours during mechanical ventilation. Given its beneficial sedative and anxiolytic properties and limited adverse effect profile, it has been used in several other clinical scenarios. The authors present their experience using dexmedetomidine for monitored anesthesia care (MAC) during "awake" ENT procedures such as thyroplasty, a procedure requiring a patient to verbalize when requested but to otherwise remain immobile to allow for completion of the procedure, and in a patient with post-polio syndrome with poor pulmonary reserve requiring esophagoscopy with dilation and botulinum toxin injection for cricopharyngeal dysfunction. Our preliminary experience suggests that dexmedetomidine provides effective sedation as the primary agent for MAC during such procedures in adult patients. The end-organ effects of dexmedetomidine and previous reports of its use during MAC are reviewed. PMID- 19127136 TI - Prevalence of influenza vaccination and pneumococcal vaccination in elderly and high-risk patients seen in a university general medicine clinic. AB - Charts of 240 unselected patients (149 women and 91 men), mean age 74 +/- 7 years (range, 64-95 years), seen in a university general medicine clinic at Westchester Medical Center/New York Medical College between April 2004 and April 2007 were reviewed for the use of influenza vaccination and pneumococcal vaccination. Of the 240 patients, 108 (45%) had cardiovascular disease, 183 (76%) had hypertension, 70 (29%) had diabetes mellitus, 32 (13%) had chronic pulmonary disease, 30 (13%) had cancer, 26 (11%) had chronic renal disease, and 19 (8%) had no chronic illness. Of the 240 patients, 24 (10%) refused influenza vaccination and two (1%) had a hypersensitivity to eggs. Of 240 patients, 18 (8%) refused pneumococcal vaccination. Of 94 patients who did not refuse influenza vaccination or were allergic to eggs during the winter season of October 2004 through January 2005, 58 (62%) had influenza vaccination. Ninety-two of 172 patients (54%) had influenza vaccination during October 2005 through January 2006. Ninety-seven of 136 patients (71%) had influenza vaccination during October 2006 through January 2007. Of 222 patients, 111 (50%) had pneumococcal vaccination during 2004 through 2007. PMID- 19127137 TI - Cocaine-induced chest pain and beta-blockade: an inner city experience. AB - BACKGROUND: Cocaine is the most common illicit drug used in patients presenting with chest pain to emergency departments. Data on beta-blockers in cocaine related chest pain syndrome are sparse. We sought out to study the causal and detrimental effects of beta-blockers in cocaine-related chest pain in a large inner city cohort of patients. METHODS AND RESULTS: All patients presenting to a large inner city emergency department with chest pain, with positive urine drug screen for cocaine were included. The group comprised predominantly young (mean age 46.8 +/- 8.2 years), African American (90.6%) males (73.4%). Evidence of myocardial infarction in the form of elevation of troponin-I was noted in 7.3%. Evidence of myonecrosis (MN) was significantly more likely in those who were taking beta-blockers at presentation as compared with those who were not (14% versus 4.4%, P < 0.01). In the absence of prospective controlled data, our observational findings seem to suggest that routine initiation or continuation or of beta-blockers after admission increased the likelihood of developing MN (23.3% versus 10.7%, P < 0.01) during the course of hospitalization. CONCLUSIONS: MN as reflected by elevation of cardiac biomarkers is uncommon in patients presenting with cocaine-related chest pain. Preexisting use of beta-blockers seems to render a higher risk of myocardial injury in patients presenting with cocaine-related chest pain. In addition initiation or continuation of beta-blockers during hospitalization should be discouraged. PMID- 19127138 TI - Is there a dark side to long-term proton pump inhibitor therapy? AB - Proton pump inhibitors (PPIs) are among the most widely used of prescription drugs. They have revolutionized the management of gastroesophageal reflux disease and other acid-related disorders. Although generally safe, concerns about possible adverse effects continue to arise. Some of these, such as gastric neoplasms, are of theoretical concern only and are related to suppression of gastric acid secretion and consequent hypergastrinemia; these have not been encountered in clinical practice despite millions of patient-years of use. Others are more idiosyncratic, unpredictable, and rare. In general, the therapeutic benefits of PPIs outweigh these potential risks. However, it is important that PPIs are only given for appropriate indications and that, whenever possible, they are used in the lowest effective dose. At present, there is no need for specific monitoring for adverse events during PPI therapy. PMID- 19127139 TI - Optimizing insulin therapy in patients with type 1 and type 2 diabetes mellitus: optimal dosing and timing in the outpatient setting. AB - Management of type 1 and type 2 diabetes is continually evolving, and among these evolving therapies is administration of insulin in its various forms. The insulin regimen needs to be tailored to each individual, not only to maximize compliance and glycemic control but also to minimize hypoglycemia and weight gain. PMID- 19127141 TI - beta-Adrenergic blockers: a 50-year historical perspective. AB - The development and subsequent clinical application of the beta-adrenergic receptor blocking drugs over the past 50 years represent one of the major advances in human pharmacotherapy. No other class of synthetic drugs has demonstrated such widespread therapeutic utility for the treatment of so many cardiovascular and noncardiovascular diseases. In addition, these drugs have proven to be molecular probes that have contributed to our understanding of disease, and on the molecular level, both the structure and the function of the 7 transmembrane G protein receptors, which mediate the actions of many different hormones, neurotransmitters, and drugs. The evolution of beta-blocker drug development has led to refinements in their pharmacodynamic actions that include agents with relative beta1-selectivity, partial agonist activity, concomitant alpha-adrenergic blockers activity, and direct vasodilator activity. In addition, long-acting and ultra-short-acting formulations of beta-blockers have also demonstrated a remarkable record of clinical safety in patients of all ages. Indeed, the beta-adrenergic blockers have provided us with a great clinical legacy for now and in years to come. PMID- 19127140 TI - Novel therapies for cyclic GMP control of vascular smooth muscle growth. AB - Cyclic GMP, guanosine 3',5'-cyclic monophosphate, is a critical and multifunctional second-messenger molecule that mediates diverse physiological and pathophysiological functions in cardiac and vascular tissues. Synthesized through nitric oxide, carbon monoxide, and/or natriuretic peptide-mediated guanylate cyclase stimulation and guanosine triphosphate dephosphorylation, cyclic GMP is capable of stimulating a cascade of serine/threonine kinase events, including signaling through cyclic GMP- and/or cyclic AMP-dependent protein kinases, eliciting protein kinase-independent actions such as modulation of ion channels or transporters, or undergoing hydrolytic degradation through actions of cyclic GMP-regulated phosphodiesterases. Substrates, enzymes, cofactors, and associated variables in this multifaceted system have historically been targets of vital pharmacotherapies with perhaps most common the use of vascular smooth muscle targeting organonitrates in cardiac patients and phosphodiesterase inhibitors in individuals with erectile dysfunction. Accumulating basic science and clinical evidence, however, suggests that cyclic GMP signaling is compromised under conditions of disease or elevated physiological stresses. Moreover, nitric oxide can stimulate an array of cytotoxic effects and nitric oxide-based therapies can be limited by diminished bioactivity and the development of tachyphylaxis or tolerance after prolonged use. Consequently, an emerging area for clinical drug development and therapeutic drug evaluation for conditions of cardiovascular adversity has focused on identification of cyclic GMP signaling pathways that act under oxidized or nitric oxide-unresponsive conditions and/or that operate irrespective of nitric oxide-induced complications. The aim of this therapeutic review is to describe novel, nitric oxide-alternate avenues for cyclic GMP signaling in vascular smooth muscle growth with particular emphasis on pharmacotherapeutics of recently characterized cyclic GMP-specific approaches. PMID- 19127142 TI - Motor control patterns during an active straight leg raise in pain-free subjects. AB - STUDY DESIGN: Repeated measures. OBJECTIVE: To investigate motor control (MC) patterns of normal subjects during the low level physical load of the active straight leg raise (ASLR). SUMMARY OF BACKGROUND DATA: Aberrant MC patterns, as observed with the ASLR test, are considered to be a mechanism for ongoing pain and disability in subjects with chronic musculoskeletal pelvic girdle pain. These patterns may not only affect the provision of lumbopelvic stability, but also respiration and the control of continence. Greater understanding of MC patterns in pain-free subjects may improve the management of pelvic girdle pain. METHODS.: Fourteen pain-free nulliparous women were examined during the ASLR. Electromyography of the anterior abdominal wall, right chest wall and the anterior scaleni, intraabdominal pressure (IAP), intrathoracic pressure (ITP), respiratory rate, pelvic floor kinematics, and downward leg pressure of the nonlifted leg were compared between a left and right ASLR. RESULTS: There was greater activation of obliquus internus abdominis and obliquus externus abdominis on the side of the ASLR. The predominant pattern of activation for the chest wall was tonic activation during an ipsilateral ASLR, and phasic respiratory activation lifting the contralateral leg. Respiratory fluctuation of both IAP and ITP did not differ lifting either leg. The baseline shifts of these pressure variables in response to the physical demand of lifting the leg was also the same either side. There was no difference in respiratory rate, pelvic floor kinematics, or downward leg pressure. CONCLUSION: Pain-free subjects demonstrate a predominant pattern of greater ipsilateral tonic activation of the abdominal wall and chest wall on the side of the ASLR. This was achieved with minimal apparent disruption to IAP and ITP. The findings of this study demonstrate the plastic nature of the abdominal cylinder and the flexibility of the neuromuscular system in controlling load transference during an ASLR. PMID- 19127143 TI - Biomechanical pullout strength and stability of the cervical artificial pedicle screw. AB - STUDY DESIGN: This study used cadaveric specimens to compare the biomechanical performance of artificial pedicle screws (APS) versus lateral mass screws (LMS). OBJECTIVE: The goal of this study was to biomechanically characterize APS range of-motion and pullout strength in surgical instances that preclude LMS insertion. SUMMARY OF BACKGROUND DATA: Posterior approaches used in instances of ventral spine tumors often necessitate complete facetectomy, thereby removing fixation points for LMS and requiring longer constructs with fewer segmental fixations to span the resected levels. Recently, APS were developed to overcome this obstacle. Although APS have been used successfully in clinical cases, they have yet to be biomechanically validated. METHODS: Seven fresh-frozen cervical spine segments (C2-C7) were harvested from human donors (F = 1, M = 6; 65 +/- 5 years old, range: 50-72 years old). Nondestructive range-of-motion tests were conducted on each specimen in its intact and surgically destabilized states, and after each of 3 different APS and LMS surgical stabilizations. After nondestructive bending tests, a final pullout test of APS and LMS at the C4 level was performed for each specimen. RESULTS: The pullout strength of the APS was twice as strong on average as that for the LMS (503.4 +/- 338.3 vs.- 254.3 +/- 142.3 N); this difference approached but was not statistically significant (P = 0.07). There was no significant difference in specimen stiffness between the APS- and LMS instrumented configurations in all ranges of primary and off-axis motions (P > 0.05). However, all fixation methods increased specimen stiffness in comparison with the intact conditions (1.7-36.5 times increase; P < 0.05). CONCLUSION: Our results demonstrate that APS provide comparable stability to LMS and can therefore be considered a viable alternative in surgical scenarios requiring the complete removal of lateral masses. Moreover, APS may provide some enhanced strength in the face of destructive pullout forces. PMID- 19127144 TI - Clinical significance of cervical vertebral flexion and extension spatial alignment changes. AB - STUDY DESIGN: Radiographic review. OBJECTIVE: To study changes in alignment and curvature of the cervical vertebral column in the sagittal plane, and to provide references for the orthopedic treatment of cervical kyphosis. SUMMARY AND BACKGROUND DATA: There are few reports analyzing changes of the vertebral body itself or changes in their relationships between separate vertebrae. To understand cervical vertebrae and curvature dynamic variation patterns in the sagittal plane and provide a theoretical reference for orthopedic correction of cervical kyphosis, we measured spatial position and alignment of the cervical vertebrae on sagittal plane films of various postures. METHOD: Blind repeated measures design analysis of sagittal view standard lateral, hyper flexion, and hyper extension cervical spine of 60 healthy subjects measuring spatial alignment angle of C2 to C7 vertebral bodies in the sagittal plain ( angleA), C2 to C7 inferior terminal lamina tilt angle of ( angleB), and segmental intervertebral space angles C2/3 to C6/7 ( angleC) calculating flexion and extension amplitude changes in angleA, angleB, and angleC. Cervical curve apex was determined using Borden's method to compare change and distribution characteristics. RESULTS.: Segmental angleA is a positive mean value that increases from C2 to C7 when in the upright position. In hyper extension, mean angleA value decreases with gradual amplitude decrease from C2 to C7; however, a gradual overall increasing pattern is observed for mean angleA from C2 to C7. In hyper flexion, segmental angleA mean value increases with gradual amplitude decrease from C2 to C7, whereas with decreasing angular measurements from C2 to C7. angleB follows similar change regularities as angleA with a larger mean value than angleA. During maximum extension and maximum flexion, angleD is equal to angleE. For example: C2 angleD = C2 variable angleA; C2 angleE angle(C3 variable angleA + C2/3 variable angleC); and C2 angleD = C2 angleE. Cervical spine curve apex is mainly distributed between C4 and C5 on standard, hyper extension and hyper flexion lateral view. CONCLUSION: Dynamic cervical curvature changes based on a central apex, stems from vertebral rotation and displacement in the sagittal plane. Our study reveals variation patterns of dynamic cervical spine sagittal alignment and curvature, providing vertebral spatial alignment value as reference for orthopedic cervical kyphosis corrective surgery. PMID- 19127145 TI - How often is low back pain not coming from the back? AB - STUDY DESIGN: Consecutive case series cohort. OBJECTIVE: To determine the relative frequencies of the spine, the sacroiliac (SI) joint, and the hip joint being the primary pain generator among patients presenting at a spine surgery clinic for low back pain (LBP). SUMMARY OF BACKGROUND DATA: Identification of the primary pain generator in a patient with LBP is difficult. Possible pain sources include the lumbar spine, the SI joint, and the hip joint. Their relative frequencies among patients presenting at a spine surgeon's clinic have not been well established. METHODS: Three hundred sixty-eight new patients were seen at a single spine surgeon's clinic during a 10-month period. Of these, 289 (78.5%) complained primarily of LBP with or without leg pain. Seventy-seven had previous surgery. The remaining 200 cases were reviewed for all diagnostic tests performed, as well as the final diagnosis. RESULTS: One hundred sixty-four (82%) had spine pathology, but only 130 (65%) had spine-only pathology, whereas 35 (17.5%) had a combination of spine plus hip and/or SI joint pathology. An additional 16 (8%) had hip and/or SI joint pathology without spine pathology. Twenty (10%) had an undefined pain source. Overall, 25 (12.5%) had hip pathology, and 29 (14.5%) had SI joint pathology. CONCLUSION: For patients presenting to a spine surgeon's clinic for LBP, up to 25% of patients may have significant pain contribution from the hip or SI joints, and an additional 10% will still have an undefined pain source even after diagnostic workup. This underscores the need for clinicians to be aware of nonspinal pain generators and to appropriately pursue alternative diagnoses. PMID- 19127146 TI - Single-stage posterior corpectomy and expandable cage placement for treatment of thoracic or lumbar burst fractures. AB - STUDY DESIGN: A prospective study was performed. OBJECTIVE: To assess an unusual technique for corpectomy and expandable cage placement via single-stage posterior approach in acute thoracic or lumbar burst fractures. SUMMARY AND BACKGROUND DATA: Burst fractures represent 10% to 20% of all spine injuries at or near the thoracolumbar junction, and can cause neurologic complications and kyphotic deformity. The goal of surgical intervention is to decompress the neural elements, restore vertebral body height, correct angular deformity, and stabilize the columns of the spine. METHODS: The study comprised 14 patients (8 women and 6 men aged 40.3 years) who had 1 spinal burst fracture between T8 and L4 and who underwent single-stage posterior corpectomy, circumferential reconstruction with expandable-cage placement, and transpedicle screwing between January 2003 and May 2005. Neurologic status was classified using the American Spinal Injury Association (ASIA) impairment scale and functional outcomes were analyzed using a visual analogue scale (VAS) for pain. The kyphotic angle (alpha) and lordotic angle (beta) were measured in the thoracic or thoracolumbar and lumbar regions, respectively. RESULTS.: The mean follow-up time was 24 months (range, 12-48 months). Neurologic status was in 7 patients (preop: ASIA-E, postop: unchanged), 2 patients (preop: ASIA-D, postop: 1 unchanged, 1 improved to ASIA-E), 3 patients (preop: ASIA-C, postop: 2 improved to ASIA-D, 1 improved to ASIA-E), 2 patients (preop: ASIA-B, postop: 1 improved to ASIA-C, 1 unchanged). The mean operative time was 187.8 minutes. The mean blood loss was 596.4 mL. Regarding postoperative complications, 1 patient experienced transient worsening of neurologic deficits and 1 patient developed pseudarthrosis. The mean preoperative VAS score was 8.21 and the mean postoperative VAS score was 2.66 (P < 0.05). The mean preoperative kyphotic angle for the 11 individuals with the thoracic or thoracolumbar burst fractures was 24.6 degrees and the mean preoperative lordotic angle for the 3 individuals with lumbar burst fractures was 10.6 degrees. The corresponding values at 12 months postsurgery were 17.1 degrees and 13.6 degrees. CONCLUSION: This single-stage posterior approach for acute thoracic and lumbar burst fractures offers some advantages over the classic combined anterior-posterior approach. The results from this small series suggest that a single-stage posterior approach should be considered in select cases. PMID- 19127147 TI - Ossification of ligamentum flavum related to thoracic kyphosis after tuberculosis: case report and review of the literature. AB - STUDY DESIGN: Case report and review of the literature. OBJECTIVE: To report 2 patients, with a history of thoracic tuberculosis, in whom ossification of ligamentum flavum (OLF) was found adjacent to progressive kyphosis and resulted in incomplete paralysis. The relevant literature was also to be reviewed. SUMMARY OF BACKGROUND DATA: Progressive kyphosis is the most common cause of late onset paralysis after spinal tuberculosis. However, OLF related to tuberculous kyphosis is rarely observed and this makes surgical treatment become very difficult. METHODS: Clinical history, laboratory examination, and radiographic findings of the 2 patients were described. Posterior laminectomy and instrumented fusion were performed for both 2 patients, but additional osteoectomy was performed through posterolateral zygapophysial approach in only 1 patient in whom the kyphotic deformity was partially corrected. A review of the relevant literature was also done. RESULTS: Two patients were observed up for 2 and 6 years, respectively. Neurologic status was significantly improved after operation and no further kyphosis developed. To our knowledge, only 1 article had reported 2 similar cases of spinal stenosis above a healed tuberculous kyphosis. CONCLUSION: OLF related to tuberculous kyphosis was rarely observed. The local instability and repetitive stimulus of excessive stress could lead to development of OLF. Our experience suggested that decompression of OLF was more important than correction of kyphotic deformity to improve the neurologic status in these patients. PMID- 19127148 TI - Cervical spinal meningioma mimicking intramedullary spinal tumor. AB - STUDY DESIGN: Case report. OBJECTIVE: To report a very unusual spinal meningioma, mimicking an intramedullary spinal tumor. SUMMARY OF BACKGROUND DATA: Spinal meningiomas, usually associated with signs and symptoms of cord or nerve root compression, are generally encountered in women aged over 40. Radiologic diagnosis is often established by their intradural extramedullary location on magnetic resonance images. METHODS: A 60-year-old woman had a 6-month history of progressive weakness in her upper extremities, difficulty in walking, and cervical pain radiating through both arms. Neurologic examination revealed motor strength deficiency in all her extremities, with extensor reflexes, clonus, and bilateral hyper-reflexiveness. A sensory deficit was present all over her body. Magnetic resonance images revealed that the spinal cord appeared expanded with an ill-defined, homogeneously contrast-enhanced, lobulated, eccentric mass at the C1 C3 level. The patient was operated with a preliminary diagnosis of an intramedullary tumor. RESULTS: At surgery, the mass was found to be extramedullary, and gross total resection was performed. Histopathological examination revealed a meningioma characterized by the presence of fibrous and meningothelial components. The patient was able to ambulate with a cane, and extremity strength and sensation improved 2 months after surgery. CONCLUSION: Spinal meningiomas can mimic intramedullary tumors, and should be considered in differential diagnosis of intradural tumors with atypical appearance. PMID- 19127149 TI - Aneurysmal bone cyst: surgical management in the pediatric cervical spine. AB - STUDY DESIGN: Case report. OBJECTIVE: To present 2 cases of aneurysmal bone cyst in the cervical spine of children who were aggressively treated with resection, fusion, and stabilization with hardware to achieve complete cure and future stability. SUMMARY OF BACKGROUND DATA: Aneurysmal bone cyst is a rapidly expanding and destructive bone lesion of unknown etiology. Current approaches to treatment include surgical en bloc excision, complete curettage, selective arterial embolization, radiotherapy, and intralesional injection with a sclerosing agent. Fusion and stabilization with instrumentation have often been avoided in children due to concerns over instability following future axial skeleton growth. METHODS: The authors present 2 case reports of aneurysmal bone cyst in the cervical spine of children, both demonstrating deeply involved lesions with extensive bone destruction. Both were treated aggressively with resection, fusion, and stabilization with instrumentation. Additionally, the relevant background literature is briefly discussed. RESULTS: Both patients had an uncomplicated postoperative course. At 36- and 18-month follow-up, both patients have no cervical spine instability or recurrence of tumor. CONCLUSION: Treatment of aneurysmal bone cyst in the cervical spine is challenging when it occurs in close proximity to neural and vascular structures. Future stability of the cervical spine is a concern, especially when there is significant involvement of the vertebral body and posterior elements. At 36- and 18-month follow-up, the surgical approach to aneurysmal bone cyst presented here provided these patients with complete curative treatment and preservation of cervical spine stability. PMID- 19127150 TI - Posterior migration of fusion cages in degenerative lumbar disease treated with transforaminal lumbar interbody fusion: a report of three patients. AB - STUDY DESIGN: A case report of 3 patients with posterior migration of bullet shaped fusion cages after transforaminal lumbar interbody fusion (TLIF). One patient required emergency revision surgery; the other 2 patients are being observed during conservative treatment. OBJECTIVE: To review cases of posterior migration of fusion cages and report ensuing clinical courses. SUMMARY OF BACKGROUND DATA: TLIF is a commonly used procedure; however, there are few reports describing cage migration after the procedure. In most cases, when posterior cage migration follows posterior lumbar interbody fusion, emergency revision surgery is required. One recent study reported a case of posterior cage migration after TLIF, which was treated conservatively. METHODS: Posterior migration of the bullet-shaped fusion cages occurred 1 to 2 months after TLIF in 3 patients. One of the 3 patients had isthmic spondylolisthesis treated by TLIF with bilateral pedicle screw fixation. The other 2 patients had degenerative scoliosis and were treated by TLIF with unilateral pedicle screw fixation. RESULTS: The patient with isthmic spondylolisthesis required revision surgery because the migrated cage caused nerve root irritation. The migrated cage was removed and a large-sized cage was employed to achieve stability. The other 2 patients had no pathologic symptoms after the posterior migration of the cage and were treated conservatively and observed. CONCLUSION: Revision surgery after TLIF appears relatively safe because the migrated cage tends to locate more laterally than in patients with cage migration after posterior lumbar interbody fusion. Cage migration subsequent to TLIF may not cause compression of neural tissues, so conservative treatment may suffice for these patients. Unilateral pedicle screw fixation may not provide sufficient stability to prevent cage migration in patients with degenerative scoliosis. Further study is needed to clarify surgical indications for unilateral pedicle screw fixation in TLIF. PMID- 19127151 TI - Lordoscoliosis and large intrathoracic airway obstruction. AB - STUDY DESIGN: Case series. OBJECTIVE: We report the treatment of 2 children with right main bronchus obstruction complicating thoracic lordoscoliosis. SUMMARY OF BACKGROUND DATA: The preoperative investigation and treatment of large airway obstruction caused by lordoscoliosis has not been reported in the literature. METHODS: Obstruction of the right main bronchus was confirmed before surgery by ventilation-perfusion scans, bronchogram, and computed tomography scan. Deformity correction was achieved using a submuscular growth rod construct in one child, and posterior spinal fusion in the other. Clinical examination and repeat ventilation-perfusion scans were performed 8 weeks after surgery. RESULTS: In both children, ventilation to the right "convex" lung was reestablished after surgery. Lung function improved in both patients after surgery. CONCLUSION: This is the first report of large airway obstruction associated with thoracic lordoscoliosis in which ventilation was reestablished after spinal deformity correction. Early deformity correction is indicated in such cases because of the risk of irreversible compromise to lung ventilation and perfusion. PMID- 19127152 TI - Severe cervical kyphosis complicating halo traction in a patient with Marfan syndrome. AB - STUDY DESIGN: A case report of complications from using halo traction in a patient with Marfan Syndrome (MFS). OBJECTIVE: The aim of the article is to describe a complication of halo traction in a patient with MFS. SUMMARY OF BACKGROUND DATA: Scoliosis is a common presentation in MFS and may require surgery. Halo traction is a well-recognized adjunct for correcting severe, complex, rigid scoliotic curves. No articles have been published describing complications from the use of halo traction in patients with MFS. METHODS: Review of records and radiographs. RESULTS: A patient with MFS and severe scoliosis of 145 degrees underwent an anterior and posterior release with halo-gravity traction for 2 weeks duration, followed by a posterior fusion. Her postoperative major scoliosis curve correction was stable at 26 degrees. The patient developed cervical kyphosis and neck stiffness after the traction. Ultimately, 23 months later with 66 degrees of angulation, the increasing kyphosis warranted anterior and posterior fusion. Seven months after cervical surgery, her cervical kyphosis stabilized at 22 degrees. CONCLUSION: This complication may be explained by the laxity of the connective tissue in MFS. Also, halo traction creates more tension in the cervical spine than the thoracolumbar spine. For patients with MFS and perhaps other connective tissue disorders, halo-gravity traction should be used with caution, especially when some pre-existing cervical kyphosis is present. PMID- 19127153 TI - Anterior cervical discectomy and fusion with a locked plate and wedged graft effectively stabilizes flexion-distraction stage-3 injury in the lower cervical spine: a biomechanical study. AB - STUDY DESIGN: An in vitro three-dimensional (3D) flexibility test of human C3-C7 cervical spine specimens. OBJECTIVE: To test the hypothesis that anterior cervical fusion with a wedged graft and a locked plate can effectively stabilize the cervical spine after complete anterior and posterior segmental ligamentous release. SUMMARY OF BACKGROUND DATA: Distraction-flexion Stage 3 injuries of the lower cervical spine (bilateral facet dislocations) are usually reduced under awake cranial traction. When the magnetic resonance imaging reveals a traumatic disc prolapse, anterior cervical discectomy and fusion (ACDF) is usually recommended. Most authors advise combining ACDF with posterior instrumentation to address the insufficiency of the posterior elements. However, there is clinical evidence that ACDF with a locked plate alone suffices for the treatment of these injuries, especially in young patients. Still, there are no biomechanical studies on the effect of a locked plate on the complete anterior and posterior ligamentous-deficient young cervical spine under physiologic preload. METHODS: Eight fresh frozen human lower cervical spines (C3-C7) from young donors (age, 44.5 years; range, 21-63 years) were used. A 3D flexibility test was conducted using a moment of 0.8 Nm without preload. Flexion-extension was additionally tested using a moment of 1.5 Nm under 0 and 150 N follower preload. Spines were tested first intact, then after complete C5-C6 discectomy with posterior longitudinal ligament resection and ACDF with a wedged bone graft and a rigid locked plate, and finally after complete release of the supraspinous, interspinous, and intertransverse ligaments; the facet capsules; and ligamentum flavum. RESULTS.: When tested under 0.8 Nm moment without preload, complete posterior and anterior ligamentous release did not significantly increase the ROM of the ACDF construct in flexion-extension (P > 0.025), lateral bending (P > 0.025), and axial rotation (P > 0.025). When tested under 1.5 Nm moment with or without a compressive preload, the complete posterior and anterior ligamentous release did not significantly affect the ROM of the ACDF construct (P > 0.01). The application of preload significantly reduced the motion at the C5-C6 ACDF construct with ligamentous disruption in comparison with the motion in the absence of a preload (P < 0.01). CONCLUSION: Anterior cervical fusion with a wedged graft and a rigid constrained (locked) plate can effectively stabilize the nonosteoporotic cervical spine after complete posterior element injury when excessive ROM is prevented (for example, by the use of postoperative external immobilization). Even when the construct is subjected to higher moments, adequate stability can be achieved when physiologic preload is present. Osteoporosis and lack of sufficient preload due to poor neuromuscular control may affect long-term screw stability, and additional external immobilization may be needed until fusion matures. PMID- 19127155 TI - Culture of human anulus fibrosus cells on polyamide nanofibers: extracellular matrix production. AB - STUDY DESIGN: Studies were approved by the authors' Human Subjects Institutional Review Board. Human anulus cells were tested for growth and extracellular matrix (ECM) production in vitro. OBJECTIVE: To investigate cell attachment, cell proliferation, and ECM production of human intervertebral disc anulus cells seeded onto randomly oriented electrospun polyamide nanofibers. SUMMARY OF BACKGROUND DATA: Because nanofibrillar matrices have the potential to promote microenvironments, which may mimic in vivo conditions and resemble connective tissue, their utilization opens new avenues for cell-based tissue engineering applications for disc cells. METHODS: Anulus cells were isolated from 4 cervical spine surgical disc specimens, expanded, and seeded into either routine plastic culture (control) or a nanofiber surface of randomly oriented electrospun polyamide nanofibers (Ultra-Web-coated culture dish, Corning) with a positive charge or without a charge. Cells were cultured for 9 days, digital images captured, cells harvested, embedded in paraffin, and examined for production of extracellular matrix (ECM). Additional anulus cultures were tested to quantitatively assess total proteoglycan production and cell proliferation under control or nanofiber cultures. RESULTS: Cells attached well and exhibited cell extensions within the nanofiber layers; cells on the charged nanofiber surface deposited greater amounts of chondroitin sulfate than of type II collagen than cells cultured on the uncharged nanofiber surface. Results showed that culture of anulus cells on nanofibers was permissive for secretion and assembly of type II collagen and chondroitin sulfate. Significantly greater total proteoglycan formation was present after culture on the nanofiber with added charge conditions {control, 0.6116 microg/mL +/- 0.186 [4] [mean +/- sem(n)] vs. 1.201 +/- 0.2509 [4], P < 0.05}. Cell proliferation, however, did not differ among treatment groups. CONCLUSION: Culture of anulus cells on nanofibers was found to be permissive for secretion and assembly of type II collagen and chondroitin sulfate, and culture on nanofibers with added charge significantly increased total proteoglycan production. These novel findings point to the need for further examination of nanofibrillar 3D culture of anulus cells for tissue engineering applications. PMID- 19127157 TI - A biomechanical evaluation of graded posterior element removal for treatment of lumbar stenosis: comparison of a minimally invasive approach with two standard laminectomy techniques. AB - STUDY DESIGN: A validated finite element model of the intact lumbar spine (L1-S1) was modified to study the biomechanical changes as a result of surgical alteration for treatment of stenosis at L3-L4 and L4-L5 using 2 established techniques and 1 new minimally invasive technique. OBJECTIVE: To investigate the impact of graded posterior element removal associated with new surgical techniques on postoperative segmental motion and loading in the annulus. SUMMARY OF BACKGROUND DATA: Several studies have shown that laminectomy increases and produces segmental instability unless fusion is performed. However, no data exist comparing the biomechanical impact of completely preserving the contralateral anatomy and what effect this has compared to traditional approaches. METHODS: The effect of graded removal of posterior elements because of iatrogenic change associated with the 3 approaches was investigated using an 800 N compressive preload using the follower load technique and application of 8 Nm flexion, 6 Nm extension, 4 Nm torsion, and 6 Nm lateral bending moments. RESULTS: This study shows that removal of posterior elements for treatment of stenosis at L3-L4 and L4-L5 results in increased flexion-extension and axial rotation at the surgical site. This study also shows that the segmental motion following a traditional laminectomy is greater than the minimally invasive approach in flexion, extension, left and right axial rotation. Moderate preservation of the posterior elements which occurs in the intralaminar approach generates greater segmental motion that the minimally invasive approach in extension, left and right axial rotation. CONCLUSION: Minimization of bone and ligament removal associated with minimally invasive procedures results in greater preservation of the normal motion of the lumbar spine after surgery. This study suggests that preservation of the posterior spinal elements associated with minimally invasive surgery could minimize the risk of developing de novo postoperative changes in spinal alignment and/or acceleration of facet and disc degeneration. PMID- 19127156 TI - Age-related changes in the extracellular matrix of nucleus pulposus and anulus fibrosus of human intervertebral disc. AB - STUDY DESIGN: To characterize age-related changes in the matrix of human intervertebral disc (IVD) specimens, human specimens from the third to the eighth decade of life were collected and analyzed for collagen and proteoglycan (PG) composition. OBJECTIVE: To identify age-related changes in the concentration of matrix macromolecules (collagen and PGs, including the small leucine-rich PGs biglycan, decorin, fibromodulin, and lumican) in human anulus fibrosus (AF) and nucleus pulposus (NP). SUMMARY OF BACKGROUND DATA: IVD degeneration is associated with changes in the concentration and fragmentation of matrix molecules. Deciphering age-related matrix alterations may help us to better understand the regulatory mechanisms underlying IVD degeneration. METHODS: Forty-six whole IVDs were obtained from the thoracolumbar spines (T11-L5) of humans aged between 32 and 80 years. All specimens were classified as Thompson grade 1 or 2 according to MRI criteria. Specimens were separated into (i) outer-and (ii) inner AF, and (iii) NP. DNA, collagen, and PG contents were measured using chemical assays, whereas small nonaggregating PG levels were analyzed by comparative Western blotting. RESULTS: Total PG and collagen contents in both the AF and NP consistently decreased with aging. The concentrations of small nonaggregating PGs varied. In the outer anulus, decorin levels decreased, whereas biglycan and fibromodulin levels increased with age. In the inner anulus and nucleus, biglycan demonstrated a significant increase with aging. These changes differed in most cases from those previously reported for degenerating disc tissues. CONCLUSION: Collagen and PGs appeared to undergo specific age-related changes in the human IVD. Although the total contents of these 2 families of molecules decreased during aging, individual species of small nonaggregating PGs showed species specific age-related changes. Interestingly, the level of biglycan rose and remained elevated in all 3 compartments of the disc with aging. The functional significance of these alterations is yet to be determined. PMID- 19127158 TI - Recurrence rate of lumbar disc herniation after open discectomy in active young men. AB - STUDY DESIGN: A retrospective clinical study. OBJECTIVE: To estimate the recurrence rate of lumbar disc herniation after open discectomy in active young men using survival analysis. SUMMARY OF BACKGROUND DATA: There are few reports on the recurrence rate of lumbar disc herniation in young adults, even though this age group shows a higher incidence of disc herniation than the other age groups. In addition, most of the studies on the recurrence rate of disc herniation have reported percentages without regard to the effect of the time course. METHODS: Medical records were retrospectively reviewed and phone call surveys were undertaken for 241 patients aged from 20 to 39 who had undergone open discectomies over a period of 14 years. A diagnosis of recurrence was based on the development of new symptoms and magnetic resonance imaging showing compatible lesions in the same segment as the initial diagnosis. The recurrence rate was calculated using a survival analysis based on the Kaplan-Meier product-limit method and the log-rank test was used to evaluate the effect of patient age, level of occurrence, and type of herniated disc on the recurrence rate. RESULTS: The overall recurrence rate was 7.1% (17 patients) at a mean follow-up of 8.55 years, and the cumulative survival rate was 91.5% at a follow-up of 14 years. Survival analysis estimated a higher rate of recurrence at longer follow-up, although there was no recurrence after ninth year from the primary surgery. The recurrence rate was significantly higher for protruded discs compared with other types. CONCLUSION: Survival analysis provides a more accurate estimation of true recurrence rate. Protruded discs are more likely to show recurrence than other types. PMID- 19127159 TI - Adjacent level ossification development after anterior cervical fusion without plate fixation. AB - STUDY DESIGN: A retrospective. OBJECTIVE: The proximity of the plate to the adjacent disc space appeared to be the critical determinant of adjacent level ossification development (ALOD) but we had no data on unplated arthrodesis. Without such data, we could not be positive that ALOD was a complication related to plates. SUMMARY OF BACKGROUND DATA: We previously described the incidence and timing of ALOD after anterior cervical arthrodesis and plating. METHODS: One hundred sixty-five patients (total 330 adjacent levels) who underwent anterior cervical arthrodesis without plate fixation by a single surgeon were reviewed. The average follow-up period was 28.8 months (2-9 years); ages ranged from 32 to 79 years (median 59.86). The presence and severity of ALOD was assessed on the lateral radiographs at 3, 6, 12, and 24 months after surgery and then annually and recorded into 4 grades. RESULTS: ALOD developed in 9 patients at 10 levels (5.5% of patients and 3% of levels). Eight patients had a single-level (proximal or distal) ALOD, whereas 1 patient had both ALOD. Proximal ALOD developed in 7 cases, distal in 1 case and both in 1 case. ALOD initially appeared between 6 and 12 months in all cases: 4 cases occurred at 6 months and 5 cases at 12 months. During follow-up period, the final grade of proximal ALOD was grade I for 4 levels and grade II for 4 levels and final grade of distal ALOD was grade I for 1 level and grade II for 1 level. One patient with C3-C4 arthrodesis had both ALOD with grade I proximally and grade II distally. CONCLUSION: ALOD is infrequent when certain techniques are adhered to. These include minimal stripping of the anterior longitudinal ligament and the avoidance of Caspar pins and anterior plates. It remains to be determined exactly which of these factors plays the dominant role in the production of ALOD. PMID- 19127160 TI - Cutaneous silent periods in the assessment of mild cervical spondylotic myelopathy. AB - STUDY DESIGN: Clinical, neuroradiologic, and neurophysiologic description of 21 patients with compressive cervical spondylotic myelopathy (CCSM). OBJECTIVE: To describe the utility of cutaneous silent periods (CSPs) for functional evaluation of mild CCSM. SUMMARY OF BACKGROUND DATA: Electroneurography, electromyography, and somatosensory and motor evoked potentials (SEPs, MEPs) are routinely used for comprehensive functional neurophysiological evaluation of CCSM. CSPs have been reported in various intramedullary spinal cord lesions, however, they have not been systematically studied in mild CCSM. METHODS: We investigated 21 patients with multilevel CCSM as documented by magnetic resonance imaging. We recorded CSPs in thenar muscles after noxious digit II stimulation and compared them with median and tibial nerve SEPs and MEPs obtained from abductor digiti minimi and tibialis anterior muscles. Electroneurography and electromyography were obtained in affected myotomes. RESULTS: CSP onset and end latencies were delayed, and CSP duration was shortened, in CCSM patients. CSP abnormalities were present in 17 patients of whom all, but 1 presented with intramedullary magnetic resonance imaging hyperintensity. All 11 limbs affected by hypalgesia and thermhypesthesia had abnormal CSPs, whereas no spinothalamic deficit was noted in any limb with normal CSPs. CSP onset latency was inversely correlated with JOA score and N13 amplitude, and was positively correlated with central motor conduction time to abductor digiti minimi. CSP duration was inversely correlated with central motor conduction time to tibialis anterior. Electromyographic abnormalities were found in 7 patients. CONCLUSION: We confirm the value of neurophysiological evaluation of CCSM. MEPs were more frequently abnormal than SEPs. CSP abnormalities were almost equally sensitive as upper limb MEPs, and were highly associated with spinothalamic dysfunction. The high correlation of CSP abnormalities with corticospinal tract dysfunction suggests supraspinal influence on CSPs. Our findings corroborate the utility of CSP testing in the comprehensive assessment of intramedullary spinal cord dysfunction in CCSM. PMID- 19127161 TI - Injection therapy for subacute and chronic low back pain: an updated Cochrane review. AB - STUDY DESIGN: A systematic review of randomized controlled trials (RCTs). OBJECTIVE: To determine if injection therapy is more effective than placebo or other treatments for patients with subacute or chronic low back pain. SUMMARY OF BACKGROUND DATA: The effectiveness of injection therapy for low back pain is still debatable. Heterogeneity of target tissue, pharmacological agent, and dosage, generally found in RCTs, point to the need for clinically valid comparisons in a literature synthesis. METHODS: We updated the search of the earlier systematic review and searched the Cochrane Central Register of Controlled Trials, MEDLINE, and EMBASE databases up to March 2007 for relevant trials reported in English, French, German, Dutch, and Nordic languages. We also screened references from trials identified. RCTs on the effects of injection therapy involving epidural, facet, or local sites for subacute or chronic low back pain were included. Studies that compared the effects of intradiscal injections, prolotherapy, or ozone therapy with other treatments were excluded unless injection therapy with another pharmaceutical agent (no placebo treatment) was part of one of the treatment arms. Studies about injections in sacroiliac joints and studies evaluating the effects of epidural steroids for radicular pain were also excluded. RESULTS: Eighteen trials (1179 participants) were included in this review. The injection sites varied from epidural sites and facet joints (i.e. intra-articular injections, peri-articular injections and nerve blocks) to local sites (i.e. tender-and trigger points). The drugs that were studied consisted of corticosteroids, local anesthetics, and a variety of other drugs. The methodologic quality of the trials was limited with 10 of 18 trials rated as having a high methodologic quality. Statistical pooling was not possible because of clinical heterogeneity in the trials. Overall, the results indicated that there is no strong evidence for or against the use of any type of injection therapy. CONCLUSION: There is insufficient evidence to support the use of injection therapy in subacute and chronic low-back pain. However, it cannot be ruled out that specific subgroups of patients may respond to a specific type of injection therapy. PMID- 19127163 TI - Outcome of treatment for spinal metastases using scoring system for preoperative evaluation of prognosis. AB - STUDY DESIGN: Prospective study. OBJECTIVE: To evaluate our treatment outcome for spinal metastases using our treatment strategy based on prognostic scoring system. SUMMARY OF BACKGROUND DATA: In the treatment of spinal metastases, life expectancy is most important, and our scoring system for metastatic spine tumor prognosis has been useful for such prognostic evaluation. METHODS: Conservative treatment or palliative surgery was indicated in patients with a predicted prognosis of less than 6 months or in those with multiple vertebral metastases, whereas excisional surgery was performed in patients with a predicated prognosis of 1 year or more, or with a predicted prognosis of 6 months or more, and with metastasis in a single vertebra. One hundred eighty-three patients were prospectively treated according to this principle using our prognostic scoring system, and the outcome was evaluated. RESULTS: The consistency rate between the predicted prognosis from the criteria of the scoring system and the actual survival period was high in patients within each score range (0-8, 9-11, or 12 15), 87.9% in the 183 patients. Only the palliative surgery group (n = 55) showed a significant improvement of the Barthel index between before and after treatment (P < 0.01). The mean maximum Barthel index after treatment in any modality ran parallel to the total scores of our scoring system. CONCLUSION: The prognostic criteria using our scoring system were useful for the pretreatment evaluation of prognosis irrespective of the treatment modality. In any treatment, the survival period of the patients affected the functional prognosis; therefore, it may be appropriate and realistic to select treatment methods by giving first priority to the life expectancy of patients. PMID- 19127162 TI - Comparison of magnetic resonance imaging and computed tomography in predicting facet arthrosis in the cervical spine. AB - STUDY DESIGN: Retrospective review. OBJECTIVE: To determine the ability of magnetic resonance imaging (MRI) and computed tomography (CT) to predict the presence of cervical facet arthrosis. SUMMARY OF BACKGROUND DATA: In the Food and Drug Administration Investigational Device Exemption trials of cervical disc arthroplasty (CDA), the presence of facet arthrosis on CT was a contraindication to the insertion of a CDA. Most surgeons routinely obtain an MRI, but not necessarily a CT before performing surgery in the cervical spine. We sought to determine if the MRI alone is adequate to assess for the presence of facet arthrosis. METHODS: Three experienced spine surgeons retrospectively evaluated CT scans and MRIs of the same patients, obtained within 30 days of each other in a blinded, random fashion. Reviewers graded each of the MRI and CT scan as normal or abnormal on 3 separate occasions and if the facet was abnormal, each reviewer graded the degree of arthrosis. The radiologist's evaluation for each study was compared with our results. RESULTS: Of 594 facets analyzed, 43.1% were categorized as normal on CT, and of those, MRI concordance was only 63.7% with moderate/substantial intermethod agreement. Furthermore, MRI was concordant only 15.9% of the time in patients with ankylosed facet joints on CT. CT inter-rater reliability showed substantial agreement for diagnoses of both normal and ankylosis and fair agreement for lesser degrees of facet arthrosis. MRI inter rater reliability showed fair/moderate agreement in normal and ankylosed segments and only slight agreement with lesser degrees of facet arthrosis. CT intrarater reliability showed substantial agreement in normal or ankylosed joints, but only fair agreement for all other categories; MRI showed only fair agreement. CONCLUSION: The ability of MRI to adequately determine the presence or amount of facet arthrosis is not reliable. Additionally, for abnormal facets, MRI was not reliable in adequately determining the degree of arthrosis. Our data suggest that computed tomography remains necessary in diagnosing facet arthrosis before CDA. PMID- 19127164 TI - Chronic spinal cord injury in the pediatric population: does magnetic resonance imaging correlate with the International Standards for Neurological Classification of Spinal Cord Injury examination? AB - STUDY DESIGN: Prognostic, retrospective case series. OBJECTIVE.: This study is part of a larger investigation to develop and validate a standardized and reliable method to evaluate and classify the neurologic consequence of spinal cord injury (SCI) in children. Such an instrument may also find use in the evaluation of patients with concomitant brain injury and/or cognitive impairment. We examined the relationship between the International Standards for Neurologic Classification of Spinal Cord Injury (ISCSCI) examination and magnetic resonance imaging (MRI) findings in a pediatric SCI population. SUMMARY OF BACKGROUND DATA: Recently, the reliability of the ISCSCI in young children with SCI who are unable to cognitively engage in the examination has been called into question. This has important implications as appropriate classification of these patients is necessary for prognostication, follow-up care, and appropriate placement into clinical trials. METHODS: Our longitudinal pediatric SCI database was reviewed for children with chronic SCI (>6 months), ISCSCI examinations performed by experienced testers, and adequate MRIs of the spine. ISCSCI results were correlated with MRI findings. Twenty-six subjects were identified. RESULTS: Overall, good to excellent relationships between ISCSCI neurologic level (NL) and MRI level of injury were found [Kendall tau correlation coefficient 0.90 (P < 0.001)]. The ISCSCI NL was on an average, two-thirds of a vertebral level cephalad to the center of the lesion on MRI. One child with MRI evidence of cord disruption tested incomplete at ISCSCI examination. CONCLUSION: The ISCSCI examination was found to have good to excellent relationships with MRI level in children with chronic SCI. Our results suggest MRI may be a useful adjunct for the determination of NL in children unable to participate with the examination. Our results also suggest caution in using the ISCSCI for the determination of completeness in young children. Further research into new methods (e.g., diffusion tensor imaging) to determine completeness of injury is warranted. PMID- 19127165 TI - "Ten second step test" as a new quantifiable parameter of cervical myelopathy. AB - STUDY DESIGN: A clinical and cohort study. OBJECTIVE: We developed 10 second step test as a quantifiable measure of severity in cervical compressive myelopathy. The purpose is to establish the standard value of 10 second step test and to verify its clinical effectiveness. SUMMARY OF BACKGROUND DATA: In determining the severity of cervical myelopathy, the effects of surgical intervention, or the factors that influence prognosis, it is essential to have an objective and reproducible means of measuring the patient's disability. There were only a few scales to quantify the severity of cervical compressive myelopathy. METHODS: One hundred sixty-three preoperative patients with cervical compressive myelopathy and 1200 healthy volunteers were included. Three tests were performed: the number of steps in 10 seconds, number of finger grip and release (G and R) in 10 seconds and the Japanese Orthopedic Association score for cervical myelopathy (JOA score). The study population included 99 men and 64 women with a mean age of 63.3 years (range, 33-92). RESULTS: The average number of steps in all patients was 10.7 +/- 5.5 before surgery whereas the average number of steps in the control was 19.6 +/- 3.5. The number of steps was significantly lower in patients than in control and decreased with age in both groups. Number of steps significantly correlated with the number of G and R, walking grade of JOA scores, and total JOA score. One hundred twenty-three patients were retested at 12 months after surgery. In this group, the average data of the step test were 10.4 +/- 5.9 before surgery, and 14.0 +/- 5.4 after surgery, showing significant postoperative improvement (P < 0.0001). CONCLUSION: A 10 second step test is an easily performed, quantitative task, and useful in assessing the severity of CSM. Moreover, it can be used in determining the effects of decompressive surgical treatment. PMID- 19127166 TI - Perioperative complications in revision anterior lumbar spine surgery: incidence and risk factors. AB - STUDY DESIGN: This is a retrospective review of 129 consecutive anterior lumbar revision surgeries in 108 patients. It is a single-center, multi-surgeon study. OBJECTIVE: To determine occurrence rates and risk factors for perioperative complications in revision anterior lumbar fusion surgery. SUMMARY OF BACKGROUND DATA: Although complication rates from large series of primary anterior fusion procedures have been reported, reports of complication rates for revision anterior fusion procedures are relatively rare. Concern exists chiefly about the risk to vascular and visceral structures because of scar tissue formation from the original anterior exposure. METHODS: This was a retrospective review of 129 consecutive anterior revision lumbar surgeries in 108 patients operated between 1998 and 2003. There were 40 men and 68 women. The age of patients ranged from 25 to 83 (average 50.6 years). Patients were excluded if surgery was for tumor or infection. Patients were divided into 2 groups; those with revision surgery at the same level and those with revision surgery at an adjacent level. Outcome measures included all perioperative complications. Statistical analysis included Student t test and nonparametric sign-rank. RESULTS: The number of surgical levels treated for revision was similar between the 2 groups (1 level 69%; 2 levels 19%; 3 or more levels 12%). Revision cases at the same operative level had a higher overall complication rate (42%) compared with extensions (20%; P = 0.007). This difference was primarily because of vein lacerations (23.7% vs. 3.6%, P = 0.002). There were 2 ureteral problems, both successfully salvaged. There were no arterial injuries or deaths. CONCLUSION: Complication rates for revision lumbar surgery in this series were 3 to 5 times higher than reported for primary lumbar exposures. Complication rates were significantly higher for revision anterior lumbar fusions at the same segment, which were typically in the lower lumbar spine, compared with cases involving extensions, which were typically in the upper lumbar spine. PMID- 19127167 TI - Three-dimensional classification of thoracic scoliotic curves. AB - STUDY DESIGN: Three-dimensional (3D) characterization of the thoracic scoliotic spine (cross-sectional study). OBJECTIVES: To investigate the presence of subgroups within Lenke type-1 curves by evaluating the thoracic segment indices extracted from 3D reconstructions of the spine, and to propose a new clinically relevant means (the daVinci representation) to report 3D spinal deformities. SUMMARY OF BACKGROUND DATA: Although scoliosis is recognized to be a 3D deformity of the spine its measurement and classification have predominantly been based on radiographs which are 2D projections in the coronal and sagittal planes. METHODS: Thoracic segment indices derived from 3D reconstructions of coronal and sagittal standing radiographs of 172 patients with right thoracic adolescent idiopathic scoliosis, reviewed by the 3D Classification Committee of the Scoliosis Research Society, were analyzed using the ISOData unsupervised clustering algorithm. Four curve indices were analyzed: Cobb angle, axial rotation of the apical vertebrae, orientation of the plane of maximum curvature of the main thoracic curve, and kyphosis (T4-T12). No assumptions were made regarding grouping tendencies in the data nor were the number of clusters predefined. RESULTS: Three primary groups were revealed wherein kyphosis and the orientation of the PMC of the main thoracic curve were the major discriminating factors with slight overlap between groups. A small group (G1) of 22 patients having smaller, nonsurgical (minor) curves was identified. Although the remaining patients had similar Cobb angles they were split into 2 groups (G2: 79 patients; G3: 71 patients) with different PMC (G2: 65 degrees -81 degrees ; G3: 76 degrees -104 degrees ) and kyphotic measures (G2: 23 degrees -43 degrees ; G3: 7 degrees -25 degrees). CONCLUSION: Two distinct subgroups within the surgical cases (major curves) of Lenke type-1 curves were found thus suggesting that thoracic curves are not always hypokyphotic. The ISOData cluster analysis technique helped to capture inherent 3D structural curve complexities that were not evident in a 2D radiographic plane. The daVinci representation is a new clinically relevant means to report 3D spinal deformities. PMID- 19127168 TI - Re: Patel PN, Upasani VV, Bastrom TP, et al. Spontaneous lumbar curve correction in selective thoracic fusions of idiopathic scoliosis: a comparison of anterior and posterior approaches. Spine 2008;33:1068-73. PMID- 19127169 TI - Advanced glycation end products increase transglutaminase activity in primary porcine tenocytes. AB - OBJECTIVES: Tendon abnormalities, such as increased stiffness, thickness, and excess calcification, occur commonly in patients with diabetes mellitus and cause considerable disability. These changes are frequently attributed to increased cross-linking of extracellular matrix components by advanced glycation end products (AGEs). However, cellular effects of AGEs, such as increased activity of the cross-linking transglutaminase (Tgase) enzymes, could also contribute to altered tissue biomechanics and calcification in diabetic tendons. We determined the effect of AGE-modified protein on tenocyte Tgase activity. RESEARCH DESIGN AND METHODS: Primary porcine tenocytes were exposed to N- carboxymethyl-lysine (CML)-modified type I collagen in high or normal glucose media. Protein and mRNA levels of the Tgase enzymes and Tgase activity levels were measured, as were markers of apoptosis. We also determined the effect of antioxidants on CML collagen mediated Tgase activity. RESULTS: Carboxymethyl-lysine-collagen increased Tgase activity in tenocytes 2.3- to 5.6-fold over unmodified collagen controls in both normal and high glucose media, without altering enzyme protein levels. Anti-oxidant treatment reduced the effect of CML-collagen on Tgase activity. Deoxyribonucleic acid laddering and annexin V protein levels were not altered by CML-collagen exposure. CONCLUSIONS: Carboxymethyl-lysine-collagen increased Tgase activity in tenocytes, likely posttranslationally. Increased levels of Tgase-mediated cross-links may contribute to the excess calcification and biomechanical pathology seen in diabetic tendons. PMID- 19127170 TI - Risk and resilience: early manipulation of macaque social experience and persistent behavioral and neurophysiological outcomes. AB - OBJECTIVE: To review the contributions of research on nonhuman primates, specifically macaque monkeys, to the understanding of early social stress and its effects on behavior and neurophysiology. METHOD: Review and synthesis of two bodies of work on macaque monkeys and early social manipulation: peer rearing and variable foraging demands. The literature was searched with Medline using key terms macaque, variable foraging, and peer rearing. The reference lists of these articles were also used to generate potential studies for review. RESULTS: Nonhuman primate macaques show similarities to humans in their social development and functioning. Peer rearing of young macaques and rearing of young macaques with mothers experiencing variable foraging conditions both result in increased anxious, impulsive, and aggressive temperament and behavior; more reactive stress physiology; altered neurotransmitter functioning; and immune and metabolic changes. Functional variants of specific genes that code for neuromodulators are mediators of these effects. CONCLUSIONS: Disrupted social relations during macaque rearing contribute to the risk for developing emotional and neurophysiological disturbance. In the face of such disruption, certain genotypes contribute to resilience. This can be alternately stated that, for animals of high-risk genotypes, resilience is conferred by quality relationships during rearing. This interaction of genetics with early social environment also applies to child mental health, implicating biological mediators identified in macaques as contributing to more complex outcomes in humans. PMID- 19127171 TI - Effects of MPH-OROS on the organizational, time management, and planning behaviors of children with ADHD. AB - OBJECTIVE: To evaluate the effects of stimulant medication on organizational, time management, and planning (OTMP) in children with attention deficit/hyperactivity disorder (ADHD) and ascertain whether OTMP is normalized with medication. METHOD: Participants included 19 stimulant-naive children with ADHD (aged 8-13 years) and impaired OTMP functioning, defined as greater than 1 SD below norms on the Children's Organizational Skills Scale. A double-blind, placebo-controlled, crossover design, with 4 weeks of each condition, evaluated medication (methylphenidate-osmotic-release oral system [MPH-OROS]) effects on OTMP, based on the parent and teacher versions of the Children's Organizational Skills Scale. The parent and teacher Swanson, Nolan, and Pelham, Version IV, rating scales assessed ADHD symptoms. "Not impaired" in OTMP was defined as no longer meeting study entry criteria, and "not impaired" in ADHD symptoms was defined as having mean Swanson, Nolan, and Pelham, Version IV, scores of < or = 1.0. RESULTS: MPH-OROS significantly improved children's OTMP behaviors. These improvements were correlated with significant reductions in ADHD symptoms. However, most of the children (61%) continued to show significant OTMP impairments on MPH-OROS. CONCLUSIONS: The MPH-OROS reduced children's OTMP deficits, and these improvements were associated with improvements in ADHD symptoms. Some children remained impaired in OTMP even after effective stimulant treatment of ADHD symptoms. These youngsters may require other treatments that target OTMP deficits. PMID- 19127172 TI - Remission and recovery in the Treatment for Adolescents with Depression Study (TADS): acute and long-term outcomes. AB - OBJECTIVE: We examine remission rate probabilities, recovery rates, and residual symptoms across 36 weeks in the Treatment for Adolescents with Depression Study (TADS). METHOD: The TADS, a multisite clinical trial, randomized 439 adolescents with major depressive disorder to 12 weeks of treatment with fluoxetine, cognitive-behavioral therapy, their combination, or pill placebo. The pill placebo group, treated openly after week 12, was not included in the subsequent analyses. Treatment differences in remission rates and probabilities of remission over time are compared. Recovery rates in remitters at weeks 12 (acute phase remitters) and 18 (continuation phase remitters) are summarized. We also examined whether residual symptoms at the end of 12 weeks of acute treatment predicted later remission. RESULTS: At week 36, the estimated remission rates for intention to-treat cases were as follows: combination, 60%; fluoxetine, 55%; cognitive behavioral therapy, 64%; and overall, 60%. Paired comparisons reveal that, at week 24, all active treatments converge on remission outcomes. The recovery rate at week 36 was 65% for acute phase remitters and 71% for continuation phase remitters, with no significant between-treatment differences in recovery rates. Residual symptoms at the end of acute treatment predicted failure to achieve remission at weeks 18 and 36. CONCLUSIONS: Most depressed adolescents in all three treatment modalities achieved remission at the end of 9 months of treatment. PMID- 19127173 TI - Threat-related selective attention predicts treatment success in childhood anxiety disorders. AB - OBJECTIVE: The present study examined whether threat-related selective attention was predictive of treatment success in children with anxiety disorders and whether age moderated this association. Specific components of selective attention were examined in treatment responders and nonresponders. METHOD: Participants consisted of 131 children with anxiety disorders (aged 8-16 years), who received standardized cognitive-behavioral therapy. At pretreatment, a pictorial dot-probe task was administered to assess selective attention. Both at pretreatment and posttreatment, diagnostic status of the children was evaluated with a semistructured clinical interview (the Anxiety Disorders Interview Schedule for Children). RESULTS: Selective attention for severely threatening pictures at pretreatment assessment was predictive of treatment success. Examination of the specific components of selective attention revealed that nonresponders showed difficulties to disengage their attention away from severe threat. Treatment responders showed a tendency not to engage their attention toward severe threat. Age was not associated with selective attention and treatment success. CONCLUSIONS: Threat-related selective attention is a significant predictor of treatment success in children with anxiety disorders. Clinically anxious children with difficulties disengaging their attention away from severe threat profit less from cognitive-behavioral therapy. For these children, additional training focused on learning to disengage attention away from anxiety-arousing stimuli may be beneficial. PMID- 19127175 TI - Stretch-induced strength deficits are likely significant. PMID- 19127177 TI - American College of Sports Medicine Position Stand. Appropriate physical activity intervention strategies for weight loss and prevention of weight regain for adults. AB - Overweight and obesity affects more than 66% of the adult population and is associated with a variety of chronic diseases. Weight reduction reduces health risks associated with chronic diseases and is therefore encouraged by major health agencies. Guidelines of the National Heart, Lung, and Blood Institute (NHLBI) encourage a 10% reduction in weight, although considerable literature indicates reduction in health risk with 3% to 5% reduction in weight. Physical activity (PA) is recommended as a component of weight management for prevention of weight gain, for weight loss, and for prevention of weight regain after weight loss. In 2001, the American College of Sports Medicine (ACSM) published a Position Stand that recommended a minimum of 150 min wk(-1) of moderate-intensity PA for overweight and obese adults to improve health; however, 200-300 min wk(-1) was recommended for long-term weight loss. More recent evidence has supported this recommendation and has indicated more PA may be necessary to prevent weight regain after weight loss. To this end, we have reexamined the evidence from 1999 to determine whether there is a level at which PA is effective for prevention of weight gain, for weight loss, and prevention of weight regain. Evidence supports moderate-intensity PA between 150 and 250 min wk(-1) to be effective to prevent weight gain. Moderate-intensity PA between 150 and 250 min wk(-1) will provide only modest weight loss. Greater amounts of PA (>250 min wk(-1)) have been associated with clinically significant weight loss. Moderate-intensity PA between 150 and 250 min wk(-1) will improve weight loss in studies that use moderate diet restriction but not severe diet restriction. Cross-sectional and prospective studies indicate that after weight loss, weight maintenance is improved with PA >250 min wk(-1). However, no evidence from well-designed randomized controlled trials exists to judge the effectiveness of PA for prevention of weight regain after weight loss. Resistance training does not enhance weight loss but may increase fat-free mass and increase loss of fat mass and is associated with reductions in health risk. Existing evidence indicates that endurance PA or resistance training without weight loss improves health risk. There is inadequate evidence to determine whether PA prevents or attenuates detrimental changes in chronic disease risk during weight gain. PMID- 19127178 TI - Influence of feedback and prior experience on pacing during a 4-km cycle time trial. AB - PURPOSE: To determine the importance of distance knowledge, distance feedback, and prior experience on the setting of a pacing strategy. METHODS: Eighteen well trained male cyclists were randomly assigned to a control (CON) group or an experimental (EXP) group and performed four consecutive 4-km time trials (TT), separated by a 17-min recovery. The CON group received prior knowledge of distance to be cycled and received distance feedback throughout each TT; the EXP group received neither but knew that each TT was of the same distance. RESULTS: The EXP group was significantly slower than the CON group to complete TT1 (367.4 +/- 21 vs 409.4 +/- 45.5 s, P < 0.001). Differences between groups in completion time reduced over successive TT (CON TT4 = 373.9 +/- 20 s vs EXP TT4 = 373.8 +/- 14.4 s), shown by a significant linear contrast (F1,16 = 12.39, P < 0.0005). Mean speed and power output also showed significantly reduced differences between groups over successive TT (P < 0.0005). However, peak power output showed no such convergence between groups over TT. End blood lactate was significantly different between groups in TT1, but differences between groups converged with successive TT. CONCLUSION: The progressively improving completion times in the EXP group show that distance feedback is not essential in developing an appropriate pacing strategy. Prior experience of an unknown distance appears to allow the creation of an internal, relative distance that is used to establish a pacing strategy. PMID- 19127179 TI - Effects of static stretching on repeated sprint and change of direction performance. AB - PURPOSE: To examine the effects of static stretching during the recovery periods of field-based team sports on subsequent repeated sprint ability (RSA) and change of direction speed (CODS) performance. METHODS: On four separate occasions, 12 male team-sport players performed a standardized warm-up, followed by a test of either RSA or CODS (on two occasions each) in a counterbalanced design. Both tests involved three sets of six maximal sprint repetitions, with a 4-min recovery between sets. During the break between sets, the participants either rested (control [CON]) or completed a static stretching protocol (static stretch [SS]). The RSA test involved straight-line sprints, whereas the CODS test required a change of direction (100 degrees) every 4 m (total of four). Mean, total (sum of six sprints), first, and best sprint times (MST, TST, FST, and BST, respectively) were recorded for each set. RESULTS: There was a consistent tendency for RSA times to be slower after the static stretching intervention, which was supported by statistical significance for three performance variables (MST 0-5 m set 2, MST 0-20 m set 2, and TST set 2; P < 0.05). This tendency was also supported by moderate effect sizes and qualitative indications of "likely" harmful or detrimental effects associated with RSA-SS. Further, sprint times again tended to be slower in the CODS-SS trial compared with the CODS-CON across all sprint variables, with a significantly slower (P < 0.05) BST recorded for set 3 after static stretching. CONCLUSION: These results suggest that an acute bout (4 min) of static stretching of the lower limbs during recovery periods between efforts may compromise RSA performance but has less effect on CODS performance. PMID- 19127180 TI - Pelvic stabilization and semisitting position increase the specificity of back exercises. AB - PURPOSE: To examine the effect of pelvic stabilization and two lower-limb configurations on the EMG activity of back and hip extensor muscles during a dynamic back extension exercise on a machine and to compare them between sexes. METHODS: Twenty-two healthy volunteers (11 men and 11 women) performed five trunk flexion-extension cycles at 40% of their maximal voluntary contraction (MVC) in a machine designed for back exercise. Two different positions were used: 1) seated position (seat horizontal, knees at 90 degrees ) and 2) semisitting position (seat slightly tilted forward at 15 degrees , knees at 45 degrees of flexion). In each position, three pelvic stabilization conditions were tested: 1) unrestrained (control condition), 2) partially restrained (posteriorly), and 3) totally restrained (posteriorly and anteriorly). EMG signals were recorded bilaterally with 12 pairs of electrodes placed on back muscles as well as on the gluteus maximus and biceps femoris. The muscular activation level, that is, the percentage of EMG amplitude relative to the maximal EMG obtained from MVC, was used to asses the relative contribution of each muscle group across exercises. RESULTS: In both sexes, two main results were found: 1) pelvic stabilization (partially and totally restrained) significantly (P < 0.05) increased the activity of all back muscles (average of 12%) and 2) semisitting position significantly decreased (range = 8-27%) the activity of two hip extensors compared with the seated position. Sex differences were also observed relative to the activity of some back muscles as well as the biceps femoris. CONCLUSIONS: Combining pelvic stabilization and semisitting position in back exercise machines might be a useful way to localize the effects of endurance training at the back muscles and this in both the sexes. PMID- 19127181 TI - Mechanical external work and recovery at preferred walking speed in obese subjects. AB - PURPOSE: The aim of this study was to compare the mechanical external work (per kg) and pendular energy transduction at preferred walking speed (PWS) in obese versus normal body mass subjects to investigate whether obese adults adopt energy conserving gait mechanics. METHODS: The mechanical external work (Wext) and the fraction of mechanical energy recovered by the pendular mechanism (Rstep) were computed using kinematic data acquired by an optoelectronic system and were compared in 30 obese (OG; body mass index [BMI] = 39.6 +/- 0.6 kg m(-2); 29.5 +/- 1.3 yr) and 19 normal body mass adults (NG; BMI = 21.4 +/- 0.5 kg m(-2); 31.2 +/- 1.2 yr) walking at PWS. RESULTS: PWS was significantly lower in OG (1.18 +/- 0.02 m s(-1)) than in NG (1.33 +/- 0.02 m s(-1); P beta > 0.37, all P < 0.05) and partly with speed, strength, and coordination (0.41 > beta > 0.18, all P < 0.05). In overweight children and in children from overweight parents and migrant families, this association was not found. There was no association between SCP and fatness in any of the groups. CONCLUSIONS: SCP rates were high and were associated with higher levels of most fitness components in children. Participation rates were lower for children of migrant families and children from inactive parents. In addition, the association between SCP and fitness components was not found in overweight children and in children from overweight parents and migrant families. PMID- 19127192 TI - Physical activity questionnaire comprehension: lessons from cognitive interviews. AB - PURPOSE: To determine whether respondents share researchers' understandings of concepts and questions frequently used in the assessment of usual physical activity (PA) behavior. METHODS: As part of On the Move, a study aimed at reducing measurement error in self-reported physical activity (PA), we conducted cognitive interviews with 19 men and 21 women, ages 45-65, regarding their responses to the PA questionnaires used in two large, population-based studies, Life After Cancer Epidemiology and California Men's Health study. One questionnaire asks about the frequency, the duration, and the perceived intensity of a range of specific activities in several different domains over the past 12 months. The second questionnaire asks about frequency and duration of specific, mostly recreational activities, grouped by intensity (i.e., moderate or vigorous) over the past 3 months. We used verbal probing techniques to allow respondents to describe their thought processes as they completed the questionnaires. All interviews were tape-recorded and transcribed, and the transcripts were then analyzed using standard qualitative methods. RESULTS: Cognitive interviews demonstrated that a sizable number of respondents understood "intensity" in terms of emotional or psychological intensity rather than physical effort. As a result, the perceived intensity with which a participant reported doing a specific activity often bore little relationship to the MET value of that activity. Additionally, participants often counted the same activity more than once, overestimated work-related PA, and understood activities that were grouped together in a single category to be definitive lists rather than examples. CONCLUSION: Cognitive interviews revealed significant gaps between respondents' interpretations of some PA questions and researchers' assumptions about what those questions were intended to measure. Some sources of measurement error in self-reported PA may be minimized by additional research that focuses on the cognitive processes required to respond to PA questionnaires. PMID- 19127193 TI - Self-reported physical activity improves prediction of body fatness in young adults. AB - PURPOSE: To examine self-reported physical activity levels from the International Physical Activity Questionnaire (IPAQ) as an independent predictor of dual-energy x-ray absorptiometry (DXA)-measured percent body fat (%BF) from body mass index (BMI), gender, and race. METHODS: Two hundred and seventy-eight students, aged 18 24 yr, volunteered to participate. There were 133 males (85 white and 48 black) and 145 females (77 white and 68 black). Total activity levels were quantified in MET hours per week (MET h wk(-1)) using the IPAQ short form. Height and weight were measured, and BMI values were calculated (kg m(-2)). %BF was assessed using DXA. Linear regression analysis was used to develop and compare a body fat prediction equation with (full) and without (reduced) the variable MET-h.wk. Both models included BMI, gender, and race as predictor variables. The prediction sum of squares (PRESS) statistic was used to cross-validate both models, and the individual predictive accuracy was compared using modified Bland-Altman plots. RESULTS: Mean +/- SD values were as follows: BMI = 24.4 +/- 4.1 kg m(-2), %BF = 24.5 +/- 9.3%, and MET h wk(-1) = 37.4 +/- 21.9. Gender, BMI, and race explained 81% of the variance in %BF, with a root mean square error (RMSE) of 4.07. The full model with MET-h.wk improved the prediction of %BF by 2% (R2 = 0.83, RMSE = 3.87). When cross-validated, the corresponding PRESS statistics for the reduced and full model were 4.10 and 3.90, respectively. Bland-Altman limits of agreement were greater for the reduced model compared with the full model (-8.09, 8.10 vs 7.67, 7.68). CONCLUSION: These results suggest that %BF can be predicted with greater precision and accuracy in a young adult population when MET-h.wk are included in addition to BMI, gender, and race. PMID- 19127194 TI - Monthly variation in physical activity levels in postmenopausal women. AB - INTRODUCTION: Month-to-month variation in physical activity levels in a cohort of postmenopausal women participating in a single site clinical trial undergoing lifestyle intervention was investigated before and after lifestyle intervention. METHODS: Participants were Caucasian and African American women (mean age = 57.0 +/- 3.0 yr) from the Women on the Move through Activity and Nutrition study. Physical activity was measured subjectively by questionnaire (past week and past year) and objectively by pedometer at the baseline and at the 18-month follow-up. RESULTS: At baseline, before intervention, pedometer steps were highest in the summer months (7616 steps per day), lower in the fall (6293 steps per day), lowest in winter (5304 steps per day), and then rebounded in the spring (5850 steps per day). Physical activity estimates from the past-week subjective measure followed the same seasonal pattern. After 18 months, the lifestyle change group significantly increased their pedometer step counts when compared with the health education group (P < 0.0001). At 18 months, pedometer step counts for the health education group appeared to fluctuate from month to month, whereas month-to-month step counts for the lifestyle change group appeared to remain consistent throughout the year. CONCLUSIONS: These results confirm previous reports that suggest physical activity levels fluctuate throughout the year. Lifestyle intervention, which includes a physical activity component, not only increases step counts but appears to reduce some of variation in physical activity levels over the course of a year in postmenopausal women. PMID- 19127195 TI - Age differences in the relation of perceived neighborhood environment to walking. AB - PURPOSE: The strength of the relationship of environment to physical activity may differ by age group. Older adults were expected to be more affected by environmental attributes than younger adults. The present study examined age related differences in associations between perceived neighborhood environment and physical activity. METHODS: Participants were 1623 adults aged 20 to 97 yr divided into five groups: ages 20-39, 40-49, 50-65, 66-75, and 76+. They were recruited from King County/Seattle, WA, neighborhoods selected to vary in land use and median income. Participants completed questionnaires about neighborhood environment attributes and walking for transportation and for leisure purposes. Neighborhood environment, within a 15- to 20-min walk from home, was measured on nine attributes with the validated Neighborhood Environment Walkability Scale questionnaire: residential density, proximity to nonresidential land uses, ease of access to nonresidential uses, street connectivity, walking/cycling facilities, esthetics, pedestrian traffic safety, crime safety, and proximity to recreation facilities. Participants reported frequency and duration of walking using the validated International Physical Activity Questionnaire and the Community Healthy Activities Model Program for Seniors. Partial correlations were computed, adjusting for demographics. RESULTS: Walking for transportation was significantly related to multiple perceived neighborhood attributes in all age groups, although walking for leisure was not. Walking for transportation was significantly related to almost all neighborhood environment variables in the youngest age group. In contrast, only two environmental attributes, proximity to nonresidential uses (like shops) and recreation facilities, were moderately correlated with walking for transportation in the two oldest groups. CONCLUSION: Communities need to be designed with many favorable environmental attributes to support walking for transportation among younger adults. Having nonresidential destinations and recreation facilities within walking distance may be among the most important attributes to support older adults' physical activity. PMID- 19127196 TI - Does padded headgear prevent head injury in rugby union football? AB - BACKGROUND: Concussion is a serious problem in many contact sports, including rugby union football. The study's primary aim was to measure the efficacy of padded headgear in reducing the rates of head injury or concussion. METHODS: A cluster randomized controlled trial with three arms was conducted with rugby union football teams as the unit of randomization. Teams consisted of males participating in under 13-, 15-, 18-, and 20-yr age group competitions. The interventions were "standard" and "modified" padded headgear. Headgear wearing and injury were measured for each study team at each game over two seasons. RESULTS: Eighty-two teams participated in year 1 and 87 in year 2. A total of 1493 participants (10,040 player hours) were in the control group, 1128 participants (8170 player hours) were assigned to the standard headgear group, and 1474 participants (10,650 player hours) were assigned to the modified headgear group. The compliance rates were low in all groups, but 46% of participants wore standard headgear. An intention-to-treat analysis showed no differences in the rates of head injury or concussion between controls and headgear arms. Incidence rate ratios for standard headgear wearers referenced to controls were 0.95 and 1.02 for game and missed game injuries. Analyses of injury rates based on observed wearing patterns also showed no significant differences. Incidence rate ratios for standard headgear wearers referenced to nonwearers were 1.11 and 1.10 for game and missed game injuries. CONCLUSIONS: Padded headgear does not reduce the rate of head injury or concussion. The low compliance rates are a limitation. Although individuals may choose to wear padded headgear, the routine or mandatory use of protective headgear cannot be recommended. PMID- 19127197 TI - Exercise decreases the risk of metabolic syndrome in elderly females. AB - PURPOSE: To determine the effect of exercise based on a multiple purpose, high intensity philosophy on parameters of metabolic syndrome (MetS) in older females with the MetS. METHODS: Thirty-two women (68.7 +/- 3.4 yr) with the MetS according to the International Diabetes Federation (IDF) took part in the exercise training (EG), and 33 females (69.5 +/- 4.3 yr) also with MetS served as control group (CG). Beside the diagnostic tools of the MetS (waist circumference, triglycerides, HDL-C, blood pressure, and glucose), corresponding anthropometric parameters, total cholesterol, and high-sensitivity C-reactive protein (hsCRP) were determined. RESULTS: After 12 months of exercise, significant effects were observed for total body fat [EG: -1287 g vs CG: +484 g; P = 0.001; confidence interval (CI) = -2787 to -744 g], trunk fat (-1070 vs -203 g; P = 0.005; CI = 1460 to -274 g), hip circumference (-2.1 vs +1.0 cm; P = 0.001; CI = -4.8 to -1.3 cm), triglycerides: (-24.1 vs -0.3 mg dL(-1); P = 0.021; CI = -43.9 to -3.7 mg dL(-1)), total cholesterol: (-19.8 vs +0.2 mg dL(-1); P = 0.008; CI = -34.6 to 5.5 mg dL(-1)), and HDL-C: (+3.8 vs -0.2 mg dL(-1); P = 0.036; CI = 0.3 to 7.6 mg dL(-1)). Waist circumference slightly decreased in both groups (EG: -0.7% vs CG: 0.6%; P = 0.94). Resting systolic (-7.1 vs -6.4 mm Hg; P = 0.84) and diastolic ( 8.0 vs -+9.8 mm Hg; P = 0.39) blood pressure significantly decreased in both groups. Changes of glucose and hsCRP did not significantly vary between groups (glucose: +0.7 vs +1.0 mg dL(-1), P = 0.88; hsCRP: -0.68 vs -0.50 mg L(-1), P = 0.36). The number of criteria of the MetS significantly decreased in the EG (4.13 3.66) and slightly decreased in the CG (4.12-3.97); however, significant time group interactions were not observed (P = 0.15; CI = -0.74 to 0.12). CONCLUSION: Our multipurpose exercise program significantly affects most parameters of the MetS in elderly women. PMID- 19127198 TI - Bone density comparisons in male competitive road cyclists and untrained controls. AB - Low bone mineral density (BMD) has been documented in endurance-trained runners; however, the bone status of cyclists is unclear. PURPOSE: The purpose of this study was to compare total body, lumbar spine, and dual proximal femur BMD in male competitive road cyclists (n = 32) and in age- and body mass-matched controls (n = 30). METHODS: The cyclists had an average of 9.4 yr of racing experience and trained 7-22 h wk(-1). BMD was measured using dual energy x-ray absorptiometry (DXA; GE Lunar Prodigy, v. 6.70.021). Calcium intake was estimated from a food frequency questionnaire. Resting serum total and free testosterone levels were measured by RIA (Diagnostic Systems Laboratory). RESULTS: There were no significant differences (P > 0.050) between the cyclists (CYC) and the controls (CON) for age, height, body mass, or testosterone levels. CYC had significantly (P < 0.050) lower percent body fat and higher bone-free lean body mass than the CON. Calcium intake for CYC was significantly higher (P < 0.050) than for the CON group (1557 +/- 132 vs 1098 +/- 69 mg d(-1)). Anterior-posterior spine (L2-L4) and lateral spine (LS) BMD (g cm(-2)) were significantly lower (P < 0.050) for CYC (L2-L4 = 1.165 +/- 0.023 g cm(-2); LS = 0.781 +/- 0.025 g cm(-2)) than for CON (L2-L4 = 1.246 +/- 0.028 g cm(-2); LS = 0.911 +/- 0.027 g cm(-2)). Based on t-scores (SD from the young adult reference population mean), 9% of CYC and 3% of CON were classified as osteoporotic, whereas 25% and 10% of CYC and CON, respectively, were osteopenic. CONCLUSIONS: Our findings indicated that male cyclists had lower spine BMD than controls, which was not associated with group differences in testosterone. Future studies are needed to elucidate the underlying mechanisms for low bone mass in cyclists. PMID- 19127199 TI - Responses of lean and obese boys to repeated summer exercise in the heat bouts. AB - PURPOSE: To determine the degree of natural acclimatization and artificially induced acclimation-related changes during repeated exercise in the heat bouts in seven lean and seven obese 9- to 12-yr-old boys during summer months. METHODS: Beginning at random times during the summer, subjects underwent a 70-min exercise (30% VO(2max)) in the heat exposure (38 degrees C, 50% relative humidity) on six separate days. RESULTS: On day 1, obese children were less naturally acclimatized as indicated by significantly higher baseline core temperatures (T(c)) (obese = 37.62 +/- 0.06 vs lean = 37.41 +/- 0.06; P < 0.004). By day 6 versus day 1, significant reductions in baseline T(c) were evident in both groups (obese = 37.41 +/- 0.04 vs lean = 37.18 +/- 0.04; both P < 0.05). Baseline T(c) in obese subjects by day 6 was similar to that of lean subjects on day 1. Daily reductions in exercise T(c) were evident in both groups (final exercising T(c) day 1 vs day 6: obese = 38.15 +/- 0.05 vs 37.89 +/- 0.05; lean = 38.17 +/- 0.09 vs 37.72 +/- 0.06 degrees C; both P < 0.001), occurring at a significantly slower rate in obese subjects (final exercise T(c) day 6 - day 1: obese vs lean = -0.26 +/- 0.04 vs -0.45 +/- 0.08 degrees C; P < 0.05). Significant reductions in exercising heart rate (HR) occurred in the lean but not the obese subjects by day 6 (final exercising HR day 1 vs day 6: obese = 132 +/- 3 vs 131 +/- 3, P > 0.05; lean = 138 +/- 3 vs 127 +/- 3 bpm; P < 0.001). CONCLUSIONS: During summer months, obese children are less naturally heat-acclimatized and subsequently acclimate at a slower rate. PMID- 19127200 TI - Substrate utilization during and after exercise in mild cystic fibrosis. AB - PURPOSE: To determine substrate utilization and energy expenditure during maximal and submaximal exercise and recovery in adolescents with cystic fibrosis (CF) and healthy age-matched controls (C). METHODS: Ten clinically stable CF patients (four girls, six boys; age = 10-22 yr) were matched by body mass index, age, gender, and Tanner stage to healthy controls. Subjects completed VO(2peak) testing and submaximal exercise (20 min) on a cycle ergometer at a relative intensity of 50% VO2(peak) and at an absolute power output (PO). Metabolic parameters were assessed during exercise and recovery (20 min). RESULTS: Similar respiratory quotient (RQ) values occurred in both groups during maximal exercise and recovery, despite differences in the maximal PO [CF = 114 (60-180) W and C = 171 (105-280) W, P = 0.006], the total work completed [CF = 27 (9.0-54.3) kJ and C = 55 (25.3-126.5) kJ, P = 0.008], or the VO(2peak) value attained [CF = 30.6 (8.5-45.2) mL kg(-1) min(-1) and C = 40.6 (29-64.5) mL kg(-1) min(-1), P = 0.027]. Submaximal exercise at the same absolute PO resulted in similar RQ values during exercise and recovery despite higher heart rates and average VO2 [CF = 18.8 (9.3-28.7) mL kg(-1) min(-1) and C = 15.2 (6.5-20.1) mL kg(-1)min(-1), P = 0.031] values in CF adolescents, and submaximal exercise at the same relative intensity also resulted in similar RQ values despite significantly greater average PO in the C group [CF = 38.7 (12.3-80) W and C = 67.8 (25.5-140) W, P = 0.039]. Excess postexercise oxygen consumption (EPOC) was greater in CF [2.79 (1.14-5.24) L O2] than C [1.46 (0.56-2.80) L O2] after submaximal exercise at a fixed PO (P = 0.036) but not after the relative exercise bout. CONCLUSIONS: Habitual physical activity participation does not warrant adjustment of macronutrient intake ratios in adolescents with mild to moderate CF, but total caloric intake may need to be increased based on the level of EPOC and upon the intensity and the duration of the habitual activity. PMID- 19127201 TI - Improvements in insulin sensitivity are blunted by subclinical hypothyroidism. AB - PURPOSE: Exercise- and weight loss-induced improvements in insulin resistance (IR) are variable; some individuals experience robust enhancements in insulin sensitivity, whereas others do not. Thyroid hormone status is related to IR, but it is not clear whether subclinical hypothyroidism may help to explain the variability in improvements in IR with diet and exercise. The purpose of this study was to examine whether thyroid hormone status is related to the improvement in insulin sensitivity and physical fitness after weight loss and exercise training. METHODS: By retrospective nested case-control analysis, eight subclinical hypothyroid (sHT) subjects and eight matched euthyroid controls underwent a euglycemic hyperinsulinemic clamp and peak oxygen uptake test, before and after a 16-wk program of moderate aerobic exercise combined with diet-induced weight loss. All subjects were middle-aged (57.3 +/- 3.3 yr), were overweight to obese (body mass index = 33.1 +/- 0.8 kg m(-2)), and had impaired glucose tolerance. RESULTS: The improvement in insulin sensitivity was significantly lower (P < 0.05) in the sHT group than in the euthyroid group. Both groups performed similar amounts of regular exercise and lost a significant amount of body weight during the intervention. VO(2peak) tended to improve in the euthyroid group but not in the sHT group. CONCLUSION: Subclinical hypothyroidism may interfere with beneficial adaptations on muscle metabolism and physical fitness that typically occur with weight loss and increased physical activity. These results may have significant clinical implications because of the high prevalence of both hypothyroidism and insulin resistance in the aging population. PMID- 19127202 TI - A history of medical reports on the Boston Marathon: 112 years and still running. AB - INTRODUCTION/METHODS: We performed a systematic search for medical reports on the Boston Marathon, run annually since April 19, 1897 and studied medically since 1899. RESULTS: We identified 66 articles: 25 were related to cardiology; 10, exercise physiology; 8, metabolism; 5, neurology; 4, gastroenterology; 3, hematology; 3, several disciplines; and 8, nephrology, orthopedics, and general topics. The predominance of cardiology articles reflects concerns about the cardiac risks of exercise present in the early 20th century and persistent to this day. The authors and contributors included luminaries from the medical and exercise community including Drs. Paul Dudley White, Samuel Levine, Kenneth Cooper, Paul Zoll, Ellsworth Buskirk, and David Costill. The articles identified or confirmed many of the presently accepted principles of marathon medicine. CONCLUSIONS: Medical studies on the Boston Marathon not only provide lessons applicable to managing modern athletes but also demonstrate the interests and concerns of researchers who have used the event to study the physiology of prolonged exercise for more than a century. PMID- 19127203 TI - Glucocorticoid-mediated repression of REDD1 mRNA expression in rat fetal distal lung epithelial cells. AB - REDD1 (Regulated in Development and DNA Damage-1) is a stress-response gene that represses mammalian target of rapamycin (mTOR) thus decreasing protein synthesis. In contrast to studies using cell lines and adult alveolar type II (ATII) cells, we find that REDD1 mRNA levels did not increase in rat fetal distal lung epithelia (FDLE) or fetal lung fibroblasts grown in primary cultures and then exposed to 3% O2. REDD1 mRNA expression was repressed by dexamethasone (DEX) in FDLE and ATII, but induced by DEX in fibroblasts. Lung epithelial cell lines, A549 and MLE-15, showed increases in REDD1 mRNA in response to hypoxia and DEX. The effect of DEX on REDD1 mRNA and protein in FDLE and fibroblasts was dose- and time-dependent. Inhibitor studies support repression of REDD1 mRNA by DEX in FDLE was mediated via glucocorticoid receptor and not by nongenomic effects of glucocorticoids via MAPK pathways. The half-life of REDD1 mRNA was shorter in DEX exposed FDLE compared with hormone-free media suggesting that DEX reduced REDD1 mRNA stability in FDLE. These studies indicate that REDD1 expression in response to hypoxia and DEX is cell-type specific and that physiologically appropriate levels of PO2 should be used when investigating fetal lung development. PMID- 19127205 TI - Extrauterine neuromaturation of low risk preterm infants. AB - The objective of the study was to follow neuromaturation in preterm infants. From serial exams in 90 low risk very low birthweight infants, each infant's Maturity Scores (the sum of tone, reflex, and response items) were plotted against postmenstrual age (PMA) when examined. Each infant's estimated line of best fit provides two descriptors of that infant's neuromaturation: slope (Individual Maturity Slope) and y-value (Predicted Maturity Score at 32-wk PMA). We found that Maturity Scores increased with PMA; 96% had correlation coefficients >0.8. Mean Actual and Predicted Maturity Scores at 32-wk PMA were 60 and 58, respectively, in 65 infants. When stratified by gestational age, Mean Actual Maturity Score at 30-wk PMA were 50 whether infants were 1 or several weeks old when examined. Therefore, low risk preterm infants demonstrated individual variability in rate of neuromaturation. Tone, reflexes, and responses nonetheless emerged in a predictable pattern, whether neuromaturation was intrauterine or extrauterine. This unique tool that measures preterm neuromaturation requires expertise but no technology. It has an exciting potential for providing insight into how emerging central nervous system function and structure influence each other, as well as how the central nervous system recovers from injury. PMID- 19127204 TI - Thalamic damage in periventricular leukomalacia: novel pathologic observations relevant to cognitive deficits in survivors of prematurity. AB - Despite major advances in the long-term survival of premature infants, cognitive deficits occur in 30-50% of very preterm (<32 gestational weeks) survivors. Impaired working memory and attention despite average global intelligence are central to the academic difficulties of the survivors. Periventricular leukomalacia (PVL), characterized by periventricular necrosis and diffuse gliosis in the cerebral white matter, is the major brain pathology in preterm infants. We tested the novel hypothesis that pathology in thalamic nuclei critical for working memory and attention, i.e. mediodorsal nucleus and reticular nucleus, respectively, occurs in PVL. In 22 PVL cases (gestational age 32.5 +/- 4.8 wk) and 16 non-PVL controls (36.7 +/- 5.2 wk) who died within infancy, the incidence of thalamic pathology was significantly higher in PVL cases (59%; 13/22) compared with controls (19%; 3/16) (p = 0.01), with substantial involvement of the mediodorsal, and reticular nuclei in PVL. The prevention of thalamic damage may be required for the eradication of defects in survivors with PVL. PMID- 19127206 TI - SMARCAL1 mutations: a cause of prepubertal idiopathic steroid-resistant nephrotic syndrome. AB - Schimke immuno-osseous dysplasia (SIOD) is a rare autosomal-recessive multisystem disorder with disproportionate growth failure, impaired T cell function, and steroid-resistant nephrotic syndrome. Recently, we presented the typical anthropometric features of SIOD. We now present data on two siblings who were initially classified as suffering from familial steroid-resistant nephrotic syndrome of unknown genetic origin. Apart from growth failure, no syndrome specific symptoms were found until the age of 10 y. However, serial anthropometric examinations showed the development of a SIOD-like pattern with a decreased ratio of trunk to leg length in early adolescence. The growth pattern was significantly different from that seen in children with chronic renal failure of other origins. In prepuberty the siblings had proportionate short stature but developed disproportion only during adolescence. Molecular genetic analysis revealed compound heterozygosity for a known and a new mutation in the SMARCAL1 gene. CONCLUSION: the disease spectrum associated with SMARCAL1 mutations includes previously undescribed milder phenotypes that may be clinically overlooked, particularly before puberty. Serial anthropometric assessment can eventually identify patients with a growth pattern similar to that of SIOD. These patients should be tested for SMARCAL1 mutations to avoid overtreatment with immunosuppressive agents. PMID- 19127207 TI - Plasma citrulline concentration reflects enterocyte mass in children with short bowel syndrome. AB - Plasma citrulline was recently shown to reflect the residual functional enterocyte mass in various situations characterized by intestinal failure. However, few data are available in children with short bowel syndrome. The objective of this study was to assess the value of citrulline assays in this situation. Prospective plasma citrulline assays were performed in 31 children with short bowel syndrome. Median age was 16 mo (range, 1 mo to 15 y), and median follow-up was 14 mo (6-40 mo). The energy supplied by parenteral nutrition (PN), served to assess intestinal failure severity. Plasma citrulline at inclusion showed a positive correlation with residual short bowel length. Subsequent values correlated negatively with intestinal failure severity. Plasma citrulline increased over time during or after weaning from PN (from 15.8 +/- 11.5 microM to 19.3 +/- 3.8 microM) but remained stable and low in patients who continued to need PN (6.5 +/- 3.0 microM at inclusion and 7.7 +/- 6.0 microM at last follow up). No weaned patients had a residual short bowel length less than 40 cm and plasma citrulline less than 11 microM. Our findings constitute the first evidence that serial plasma citrulline assays help to monitor residual small bowel adaptation in children. PMID- 19127208 TI - A novel mouse model of Ureaplasma-induced perinatal inflammation: effects on lung and brain injury. AB - Chorioamnionitis is associated with increased lung and brain injury in premature infants. Ureaplasma is the microorganisms most frequently associated with preterm birth. Whether Ureaplasma-induced antenatal inflammation worsens lung and brain injury is unknown. We developed a mouse model combining antenatal Ureaplasma infection and postnatal oxygen exposure. Intraamniotic Ureaplasma Parvum (UP) increased proinflammatory cytokines in placenta and fetal lungs. Antenatal exposure to UP or broth caused mild postnatal inflammation and worsened oxygen induced lung injury. Antenatal UP exposure induced central microgliosis and disrupted brain development as detected by decreased number of calbindin-positive and calretinin-positive neurons in the neocortex. Postnatal oxygen decreased calretinin-positive neurons in the neocortex but combined with antenatal UP exposure did not worsen brain injury. Antenatal inflammation exacerbates the deleterious effects of oxygen on lung development, but the broth effects prohibit concluding that UP by itself is a compounding risk factor for bronchopulmonary dysplasia. In contrast, antenatal UP-induced inflammation alone is sufficient to disturb brain development. This model may be helpful in exploring the pathophysiology of perinatal lung and brain injury to develop new protective strategies. PMID- 19127209 TI - The ratio of high-molecular weight adiponectin and total adiponectin differs in preterm and term infants. AB - Adiponectin consists of three subspecies (high-, middle- and low-molecular weight adiponectin). Among these, high-molecular weight adiponectin (H-adn) is suggested to be an active form of this protein. To assess the relationship between H-adn and postnatal growth in preterm infants (PIs), serum H-adn and total adiponectin (T-adn) were measured in 46 PIs at birth and at corrected term, and 26 term infants (TI) at birth. T-adn and H-adn concentrations, and the ratio of H-adn to T-adn (H/T-adn) were significantly greater in TI and PI at corrected term than in PI at birth (p < 0.001). T-adn and H-adn concentrations in PI at corrected term were similar to those in TI, but H/T-adn in PI at corrected term was less than that in TI (p < 0.02). Stepwise multiple regression analysis revealed that the factors contributing to H/T-adn and serum concentrations of T- and H-adn in PI at corrected term were different from those in TI. These data suggest that quality of early postnatal growth in PIs is different from that in normally developed TI. Postnatal growth accompanying adipose tissue similar to TI may be important for PI to prevent future development of cardiovascular disease. PMID- 19127210 TI - Preclinical neonatal rat studies of heparin-binding EGF-like growth factor in protection of the intestines from necrotizing enterocolitis. AB - We have previously demonstrated that enterally administered heparin-binding EGF like growth factor (HB-EGF) produced in Escherichia coli decreases the incidence and severity of intestinal injury in a neonatal rat model of necrotizing enterocolitis (NEC). In preparation for upcoming human clinical trials, large scale production of HB-EGF according to Good Manufacturing Practice (GMP) has been successfully accomplished using a Pichia pastoris yeast system. The current studies used a neonatal rat model of NEC to elucidate several important preclinical characteristics of HB-EGF therapy. We found that enteral administration of HB-EGF (800 microg/kg/dose) four times a day effectively reduced the incidence and severity of NEC, that Pichia-derived HB-EGF was not significantly different from E. coli-derived HB-EGF in preventing NEC, that EGF was not superior to HB-EGF in preventing NEC, and that prophylactic administration of HB-EGF added to formula starting with the first feed or 12 h later significantly reduced the incidence of NEC, with no change in the incidence of NEC noted if HB-EGF was added to the formula starting 24, 48, or 72 h after birth. Thus, large-scale production of GMP-grade HB-EGF in Pichia pastoris yeast produces a biologically active molecule suitable for human clinical trials. PMID- 19127212 TI - Treatment of methicillin-resistant Staphylococcus aureus in neonatal mice: lysostaphin versus vancomycin. AB - S. aureus is a significant cause of late-onset sepsis in neonates. Increasing antibiotic resistance, however, requires additional treatment options. Lysostaphin, an endopeptidase, has that potential. The objective of this study is to compare lysostaphin versus vancomycin against methicillin-resistant Staphylococcus aureus (MRSA) in a neonatal mouse model. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against MRSA strain USA300 were determined using standard methods. To determine pharmacokinetics, neonatal pups received either vancomycin or lysostaphin intraperitoneal and serum samples were obtained. To evaluate efficacy, pups were infected s.c. and littermates randomized to receive either saline, vancomycin, or lysostaphin intraperitoneal. Pups were observed for survival and growth. Quantitative blood cultures were obtained 24 h after infection. The MIC/MBC for vancomycin and lysostaphin were 0.71/1.19 microg/mL and <0.008/0.015 microg/mL, respectively. Mean lysostaphin concentrations ranged from 2.34 to 8.92 microg/mL. Mean vancomycin concentrations ranged from 1.72 to 11.2 microg/mL. Lysostaphin improved survival compared with placebo (p < 0.00001) and vancomycin (p < 0.03). There was no significant difference in growth among the groups. All treatment regimens resulted in less bacteremia compared with placebo (p < 0.0001). Lysostaphin appears to be more effective than vancomycin in treating MRSA in a neonatal model. PMID- 19127211 TI - Urinary prostasin: a possible biomarker for renal pressure natriuresis in black adolescents. AB - Prostasin is a membrane-bound/secretive serine protease interacting with aldosterone and the epithelial sodium channel in the kidney. We and others have previously proposed the concept of stress-induced pressure natriuresis (SIPN) where increased urinary sodium excretion (UNaV) is coupled with elevated blood pressure (BP) in response to behavioral stress in normotensive adolescents. This study thus aimed to test the relationship between prostasin and pressure natriuresis using the SIPN model. A cohort of 102 normotensive black adolescents (mean age: 17.0 +/- 1.2 y; 56% females) were placed on a controlled sodium (4000 +/- 200 mg/d) and potassium (2600 +/- 200 mg/d) diet for three days before testing. The SIPN protocol consisted of a 1-h baseline period, a 1-h stress period (competitive video game), and a 1-h recovery period. During the stress period, BP elevation was coupled with an increase in UNaV. Urinary prostasin concentration had more than a 2-fold reduction from baseline (38.4 +/- 32.7 ng/mL) to stress (17.2 +/- 16.0 ng/mL), and further declined during recovery (12.1 +/- 16.2 ng/mL) (p < 0.001). Urinary prostasin was inversely correlated with UNaV during stress (r = -0.43, p = 0.0001), even after being normalized by urinary creatinine. Our data suggest that urinary prostasin could be a novel biomarker and/or mechanism for renal pressure natriuresis in normotensive black adolescents. PMID- 19127213 TI - Oxygen saturation monitoring for the preterm infant: the evidence basis for current practice. AB - Many of the morbid conditions associated with extreme immaturity are potentiated by an excess of free radicals occurring in infants who developmentally have decreased levels of antioxidants. The optimal oxygen saturation values for the resuscitation, stabilization, and ongoing care of the very low birth weight infant remain largely undefined. We have reviewed the currently available evidence for clinical oxygen use in the newborn period. Until the results of further studies are available, a reasonable approach to resuscitation would include initial resuscitation with 30-40% oxygen for very preterm infants using targeted SpO2 values and blended oxygen during the first 10 min. For ongoing management of preterm infants, SpO2 targets of 85-93% seem to be most appropriate, with alarm limits set within 1 to 2% of these targets with intermittent audits to ensure compliance. There is no strong evidence to support the use of altered limits for the infant who develops early evidence of retinopathy of prematurity. Further prospective studies are required to evaluate the effects of varied oxygen targets on long-term outcome. PMID- 19127214 TI - Interleukin-10 versus dexamethasone: effects on polymorphonuclear leukocyte functions of the newborn. AB - Interleukin-10 (IL-10), an anti-inflammatory cytokine, may have therapeutic potential in the fetal inflammatory response syndrome and its sequelae such as bronchopulmonary dysplasia (BPD). Our aim was to compare the effects of IL-10 versus dexamethasone (DEX) on important PMN functions of the newborn. PMNs were isolated into culture medium from cord blood after elective cesarean section deliveries. IL-10 and DEX were compared on an equimolar basis corresponding to previously measured plasma levels of DEX from infants treated for BPD. The endotoxin (LPS)-stimulated release of the pro-inflammatory cytokines, tumor necrosis factor (TNFalpha) and IL-1 beta, were markedly inhibited equally by IL 10 and DEX; the anti-inflammatory cytokine IL-4 was not released and IL-1 receptor antagonist (IL-1ra) was released less with DEX compared with IL-10. PMNs exposed to LPS, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), or S. aureus did not show a significant difference between control, DEX and IL-10 for apoptosis, respiratory burst, phagocytosis or killing respectively. Chemotaxis to fMLP or IL-8 was unaffected by DEX or IL-10. The principal effects of both IL-10 and DEX, on the PMN functions studied, are related to the control of pro- and anti-inflammatory cytokine release. PMID- 19127215 TI - Brief bout of exercise alters gene expression in peripheral blood mononuclear cells of early- and late-pubertal males. AB - Peripheral blood mononuclear cells (PBMCs) are stimulated by exercise and contribute not only to host defense, but also to growth, repair, and disease pathogenesis. Whether PBMC gene expression is altered by exercise in children is not known. Ten early pubertal boys (8-12 y) and 10 late pubertal boys (15-18 y) performed ten 2-min bouts of strenuous, constant work rate exercise with 1-min rest intervals. PBMCs were isolated before and after exercise and microarray (Affymetrix U133 + 2 chips) analyzed. Statistical criterion to identify gene expression changes was less than 5% false discovery rate (FDR) with 95% confidence interval. One thousand two hundred forty-six genes were altered in older boys (517 up, 729 down), but only 109 were altered in the younger group (79 up, 30 down). In older boys, 13 gene pathways (using Expression Analysis Systematic Explorer, p < 0.05) were found (e.g. natural killer cell cytotoxicity, apoptosis). Epiregulin gene expression (EREG, a growth factor involved in wound healing) increased in older boys. In older boys exercise altered genes such as TBX21, GZMA, PGTDR, and CCL5 also play roles in pediatric inflammatory diseases like asthma. Sixty-six genes were changed significantly in both groups. The pattern of PBMC gene expression suggests the initiation of an immunologic "danger" signal associated with a sudden change in energy expenditure. PMID- 19127216 TI - Airway concentrations of angiopoietin-1 and endostatin in ventilated extremely premature infants are decreased after funisitis and unbalanced with bronchopulmonary dysplasia/death. AB - A systemic inflammatory response of the fetus, reflected by histologic funisitis, is a risk factor for bronchopulmonary dysplasia (BPD). Impaired pulmonary angiogenesis accompanied by simplification and rarification of alveoli is a histologic hallmark of BPD. Angiopoietin-1 mediates vascular development, maturation, and stabilization. Endostatin mainly acts as an angiostatic factor. We hypothesized that funisitis was associated with changes of endostatin and angiopoietin-1 concentrations in the airways and that an imbalance between the factors might be associated with BPD or death. We measured concentrations of angiopoietin-1 and endostatin by enzyme-linked immunosorbent assay in tracheobronchial aspirate fluid samples of 42 ventilated preterm infants during postnatal days 1 through 15. The secretory component for IgA served as reference protein. A standardized histologic examination was used to distinguish three groups: chorioamnionitis, funisitis, and controls without inflammation. Concentrations of the mediators steadily decreased. Funisitis was associated with lower concentrations of both proteins, which might impair their physiologic activities in pulmonary angiogenesis. An increase of the ratio angiopoietin 1/endostatin until day 7 of life indicated a shift of the mediators potentially favoring angiogenesis. However, infants, who developed BPD or died, had a decreased ratio on days 1, 3, and 15, suggesting an imbalance toward inhibition of pulmonary angiogenesis. PMID- 19127217 TI - Effects of different CMV-heat-inactivation-methods on growth factors in human breast milk. AB - Preterm infants can inoculate virulent cytomegalovirus (CMV) through their mothers' raw breast milk. Complete virus inactivation is achieved only by heat treatment, but the effect on growth factors has never been assessed systematically. Insulin-like-growth-factor-1-, IGF-2-, insulin-like-growth-factor binding-protein-2-, and IGFBP-3-concentrations were measured, before and after heating, in 51 breast-milk-samples from 28 mothers, and epidermal-growth-factor concentrations in a subgroup of 35 samples from 22 mothers. Two heating methods were applied: Short-term (5 s) pasteurisation at 62, 65, and 72 degrees C, and long-term Holder-Pasteurisation (30 min) at 63 degrees C. IGF-1, IGF-2, IGFBP-2, and IGFBP-3 were measured by RIA, and EGF by ELISA. Heating for 30 min decreased significantly IGF-1 by 39.4%, IGF-2 by 9.9%, IGFBP-2 by 19.1%, and IGFBP-3 by 7.0%. In contrast, IGF-1, IGF-2, IGFBP-2, and IGFBP-3 were not altered significantly when using a short heating duration of 5 s, irrespective of the level of temperature, except for IGF-2 at 62 degrees C for 5 s (p = 0.041) and IGFBP-2 at 72 degrees C for 5 s (p = 0.025). Neither long- nor short-time heating methods changed the concentration of EGF. Only short heating methods (5 s, 62-72 degrees C) can preserve, almost completely, the concentrations of IGFs in human milk, whereas Holder-Pasteurization does not. PMID- 19127218 TI - Peripheral blood flow and oxygen extraction in the sick, newborn very low birth weight infant shortly after birth. AB - This study examined the relationship between blood pressure, peripheral blood flow (PBF), and peripheral fractional oxygen extraction (FOE). Variables that may influence PBF and peripheral FOE were also measured. Measurements of PBF by near infrared spectroscopy and fractional shortening by echocardiography were made within 12 h of birth in 24 infants less than 32 wk gestation. Blood gases, Hb, temperature, and blood pressure were also measured. PBF was significantly correlated with fractional shortening (r = 0.56, p = 0.005), Po2 (r = -0.5, p = 0.01), and peripheral temperature (r = 0.52, p = 0.01). Peripheral FOE was significantly correlated with fractional shortening (r = -0.48, p = 0.02), Po2 (r = 0.52, p = 0.02), and Pco2 (r = -0.53, p = 0.008), but not with peripheral temperature. There was no significant correlation between blood pressure and either PBF or peripheral FOE. These results indicate the importance of several physiologic variables, but not blood pressure, in determining peripheral tissue oxygen delivery in sick preterm infants receiving intensive care. It adds weight to the idea that blood pressure should not be considered a surrogate for peripheral blood flow and oxygen delivery. PMID- 19127219 TI - Inhibition of JNK enhances TGF-beta1-activated Smad2 signaling in mouse embryonic lung. AB - The Smad2/3 pathway plays a key role in mediating TGF-beta1 inhibition of branching morphogenesis and induction of connective tissue growth factor (CTGF) expression in embryonic lungs. Because a number of cell-specific interactions have been described between TGF-beta1-driven Smad signaling and the c-Jun N terminal kinase (JNK) pathway, we have investigated the effects of JNK inhibition on TGF-beta1 activation of Smad2, inhibition of branching, induction of CTGF expression, and apoptosis in mouse embryonic lung explants. Mouse embryonic day 12.5 (E12.5) lung explants were treated with TGF-beta1 in the presence or absence of a specific pharmacologic JNK inhibitor (SP600125) and a specific JNK peptide inhibitor (JNKI). We found that TGF-beta1 activated the JNK pathway by stimulating c-Jun phosphorylation, which was blocked by JNK inhibitors. Treatment with SP600125 stimulated Smad2 phosphorylation and enhanced TGF-beta1-induced Smad2 phosphorylation. Treatment with JNK inhibitors also decreased normal branching morphogenesis and induced CTGF expression as well as augmented TGF beta1 inhibition of branching and induction of CTGF expression. Furthermore, JNK inhibition-induced apoptosis. Our results demonstrate that inhibition of the JNK pathway promotes TGF-beta1-driven Smad2 responses in lung branching morphogenesis. These data suggest that the JNK pathway may antagonize TGF-beta1 dependent Smad2 signaling during mouse embryonic lung development. PMID- 19127220 TI - Ibuprofen augments bilirubin toxicity in rat cortical neuronal culture. AB - Premature infants are at risk for bilirubin-associated brain damage. In cell cultures bilirubin causes neuronal apoptosis and necrosis. Ibuprofen is used to close the ductus arteriosus, and is often given when hyperbilirubinemia is at its maximum. Ibuprofen is known to interfere with bilirubin-albumin binding. We hypothesized that bilirubin toxicity to cultured rat embryonic cortical neurons is augmented by coincubation with ibuprofen. Incubation with ibuprofen above a concentration of 125 microg/mL reduced cell viability, measured by methylthiazole tetrazolium reduction, to 68% of controls (p < 0.05). Lactate dehydrogenase (LDH) release increased from 29 to 38% (p < 0.01). The vehicle solution did not affect cell viability. Coincubation with 10 microM unconjugated bilirubin (UCB)/human serum albumin in a molar ratio of 3:1 and 250 microg/mL ibuprofen caused additional loss of cell viability and increased LDH release (p < 0.01), DNA fragmentation, and activated caspase-3. Preincubation with the pan-caspase inhibitor z-val-ala-asp-fluoromethyl ketone abolished ibuprofen- and UCB-induced DNA fragmentation. The study demonstrates that bilirubin in low concentration of 10 microM reduces neuron viability and ibuprofen increases this effect. Apoptosis is the underlying cell death mechanism. PMID- 19127221 TI - Heat shock protein 70 secretion by neonatal tracheal tissue during mechanical ventilation: association with indices of tissue function and modeling. AB - Mechanical ventilation (MV) of the neonatal airway alters mechanical properties and activates tissue-modeling pathways. Heat shock protein (HSP70) is a marker of tissue injury and modulates inflammation, which may influence subsequent pulmonary tissue modeling by matrix metalloproteinases (MMPs). HSP70 secretion is up-regulated in MV airway tissues and associated with changes in airway elasticity and secretion of MMPs. Proximal tracheal segments were isolated in 13 newborn lambs and were either MV for 4 h or SHAM. At baseline and hourly, tracheal segments were flushed and tracheal elasticity was determined. Tracheal wash fluid was assayed for HSP70 by ELISA and for MMPs by substrate zymography. HSP70 secretion increased from baseline to a peak at 1 h in both groups (p < 0.01), greater in the MV group (p < 0.05), and returned to baseline values by 2 h. This response was in contrast to the progressive decrease in tracheal elasticity (p < 0.05). The HSP70 elevation pattern was noted in MMP-2, but beyond 1 h, MMP-2 returned to baseline values in MV group but remained elevated in SHAM (p < 0.05). HSP70 secretion is associated with the degree of biophysical tracheal injury as well as the time course of MMP-2 secretion by tracheal tissues. PMID- 19127222 TI - Two novel mutations in the ED1 gene in Japanese families with X-linked hypohidrotic ectodermal dysplasia. AB - X-linked hypohidrotic ectodermal dysplasia (XLHED), which is characterized by hypodontia, hypotrichosis, and hypohidrosis, is caused by mutations in ED1, the gene encoding ectodysplasin-A (EDA). This protein belongs to the tumor necrosis factor ligand superfamily. We analyzed ED1 in two Japanese patients with XLHED. In patient 1, we identified a 4-nucleotide insertion, c.119-120insTGTG, in exon 1, which led to a frameshift mutation starting from that point (p.L40fsX100). The patient's mother was heterozygous for this mutation. In patient 2, we identified a novel missense mutation, c.1141G>C, in exon 9, which led to a substitution of glycine with arginine in the TNFL domain of EDA (p.G381R). This patient's mother and siblings showed neither symptoms nor ED1 mutations, so this mutation was believed to be a de novo mutation in maternal germline cells. According to molecular simulation analysis of protein structure and electrostatic surface, p.G381R increases the distance between K375 in monomer A and K327 in monomer B, which suggests an alteration of overall structure of EDA. Thus, we identified two novel mutations, p.L40fsX100 and p.G381R, in ED1 of two XLHED patients. Simulation analysis suggested that the p.G381R mutation hampers binding of EDA to its receptor via alteration of overall EDA structure. PMID- 19127224 TI - HIV among black African communities in the UK. PMID- 19127223 TI - Force frequency relationship of the human ventricle increases during early postnatal development. AB - Understanding developmental changes in contractility is critical to improving therapies for young cardiac patients. Isometric developed force was measured in human ventricular muscle strips from two age groups: newborns (<2 wk) and infants (3-14 mo) undergoing repair for congenital heart defects. Muscle strips were paced at several cycle lengths (CLs) to determine the force frequency response (FFR). Changes in Na/Ca exchanger (NCX), sarcoplasmic reticulum Ca-ATPase (SERCA), and phospholamban (PLB) were characterized. At CL 2000 ms, developed force was similar in the two groups. Decreasing CL increased developed force in the infant group to 131 +/- 8% (CL 1000 ms) and 157 +/- 18% (CL 500 ms) demonstrating a positive FFR. The FFR in the newborn group was flat. NCX mRNA and protein levels were significantly larger in the newborn than infant group whereas SERCA levels were unchanged. PLB mRNA levels and PLB/SERCA ratio increased with age. Immunostaining for NCX in isolated newborn cells showed peripheral staining. In infant cells, NCX was also found in T-tubules. SERCA staining was regular and striated in both groups. This study shows for the first time that the newborn human ventricle has a flat FFR, which increases with age and may be caused by developmental changes in calcium handling. PMID- 19127225 TI - A new and practically challenging transition. PMID- 19127226 TI - Comparison of three peripherally-inserted central catheters: pilot study. AB - Peripherally-inserted central catheters (PICCS) are non-tunnelled, central catheters inserted through a peripheral vein of the arm. They are 50-60 cm long and are usually made of either silicone or second-third generation polyurethane. PICCs can be used for prolonged, continuous or intermittent infusion therapies (up to 3 months) both in hospitalized patients and in patients treated as outpatients, in a hospice, or at home. When establishing a vascular service it is key to select a PICC that meets the requirements of safety, cost-effectiveness, high resistance (ability to take increasing fluid volumes with high pressure devices) and durability, and low complications rate. The complications and dwell times of three different PICCs were studied: coated polyurethane, valved silicone and power-injectable. The study was conducted at the chemotherapy suite at the author's hospital with the aim of selecting the right PICC based on low incidence of complications, resistance and enhanced dwell time. Results show a low incidence of complications and long dwell time among patients with the power injectable PICC. Furthermore, this study demonstrated a reduction on the rate of occlusion and rupture with power-injectable PICCs, which makes them safer to use for administration of chemotherapy and other vesicant agents, as well as for the management of patients in critical care. PMID- 19127227 TI - Improving accuracy and efficiency of early warning scores in acute care. AB - BACKGROUND: Early warning scores (EWS) are an integral part of the care of acutely ill patients. Unfortunately, in the few studies where the accuracy of EWS has been tested it has been found to be lacking, with serious implications for quality of care. AIM: To determine if the provision of computer-aided scoring could increase the accuracy and efficiency of EWS calculations, when compared with the traditional pen-and-paper method, and to determine if it was acceptable to users. DESIGN: 26 nurses from two surgical assessment wards in two hospitals were studied. The study was conducted in three phases. Phase 1--a classroom-based exercise where nurses were given ten patient vignettes and asked to derive EWS using traditional pen-and-paper methods; Phase 2--the same as phase 1, but using a hand-held computer to derive EWS; Phase 3--the same as phase 2, but was a follow-up exercise undertaken in the ward environment, 4 weeks after computer aided scoring was implemented in the two wards. Each phase closed with a user perception/attitudes questionnaire. RESULTS: Accuracy and efficiency--phase 1 was associated with a significantly lower overall accuracy (152/260, 58%) compared with phase 2 (96%; difference in proportions 38%, 95% confidence interval 31-44%, P < 0.0001). There was a small but significant reduction in accuracy from phase 2 (96%) to phase 3 (88%) (8% difference, P=0.006). The mean time to derive an EWS reduced from 37.9 seconds in phase 1 to 35.1 seconds in phase 2 (P=0.016), down to 24.0 seconds in phase 3 (P<0.0001). User acceptability: in phase 1, nurses favoured the pen-and-paper method in all respects except accuracy. In phase 2, nurses' views shifted significantly in favour of the hand-held computer, with little deterioration in the follow-up phase 3. CONCLUSIONS: A hand-held computer helps to improve the accuracy and efficiency of EWS in acute hospital care and is acceptable to nurses. PMID- 19127228 TI - Variations in aseptic technique and implications for infection control. AB - Healthcare-acquired infections (HAIs) are a serious concern, costing the NHS 1 billion pounds a year and causing 5000 deaths annually despite increased funding. A contributing factor is the variety of aseptic techniques in use in different hospitals and even within a single hospital. These cause problems for healthcare workers as well as increasing the risk of HAI. This article examines a number of traditional approaches to aseptic technique, highlighting their differences and the implications for infection control. It concludes that improvement in aseptic technique could be achieved by implementation of a single unified approach to aseptic technique that can be standardized and audited annually, such as the aseptic non-touch technique (ANTT), which has been recommended for adoption throughout the UK. It ends with suggestions for measures that could be introduced and strengthened to improve aseptic technique, and ultimately reduce the rate of HAI. PMID- 19127229 TI - Experiences of ward-based nurse prescribers in an acute ward setting. AB - A plethora of literature is written about non-medical prescribing, directed towards community nurses, specialist or advanced practice roles. There is a lack of literature, however, regarding the use of non-medical prescribing with the open formulary in the acute ward setting. The experiences of two nurses reveal that non-medical prescribing is a valuable addition to patient care within the acute cardiac ward setting. It ensures that patients receive timely access to medications. This article reveals that the development of non-medical prescribing develops through clinical experience and with it, professional and clinical competence. Non-medical prescribing has been an invaluable addition to ward-based holistic patient care. It has ensured safety is maintained, and supported multidisciplinary working for the benefit of patients. It also provides the opportunity to develop service provision further. PMID- 19127230 TI - Specialist healthcare law for nurses: an introduction. AB - Nurses are increasingly concerned with legal and ethical dilemmas in clinical decision-making. The law impacts across a wide range of issues in nursing practice. It is important for healthcare professionals to be aware of the way in which the law regulates their professional practice. In this article John Tingle and Jean McHale introduce a new series of articles on law for nurses. The series will feature contributions by legal academics who are specialists in the area of healthcare law. The series will outline basic issues such as the English legal system. It will then consider a wide range of issues, including nursing negligence, legal issues of expanded role, patient safety, consent to treatment, adults lacking mental capacity, mental health law, end-of-life decision-making, law and nursing research, resource allocation law and human rights. PMID- 19127231 TI - Patient satisfaction with colorectal cancer follow-up system: an audit. AB - Although there have been studies into intensive follow-up versus non-intensive follow-up of patients following colorectal surgery, the evidence has not demonstrated sufficient benefit in the intensive follow-up regimen to warrant change in practice nationally, and there are no national guidelines for this. Currently at the authors' hospital the follow-up is non-intensive, and consists of an initial nurse-led surgical follow-up, dedicated patient hotline telephone number, surveillance, symptom list and open access to the nurse-led clinic as required. An audit was carried out to determine patient satisfaction with the current method of follow-up and to identify areas for improvement. The majority of patients (95%) were 'very or fairly' happy with the current service. Areas for improvement were identified. The key point from the survey was that patients would like a follow-up phone call post-diagnosis and 48 hours post-discharge from hospital. This has been implemented since the audit. PMID- 19127233 TI - A concept analysis of nurse-patient trust. AB - The aim of this article is to provide a greater understanding of the term 'trust in relation to the nurse-patient relationship through the use of Rodgers' concept analysis framework. The concept of trust is of particular interest to nursing as it has been identified as an important element in the nurse-patient relationship; however, the concept is loosely used in everyday discourse with confusion apparent and the true meaning of the concept of trust unclear. Patients' trust in the nursing profession cannot be taken for granted simply due to a requirement for nursing care. This article describes and examines the attributes, antecedents and consequences of trust in nurse-patient relationships in order to clarify the meaning and improve the current nursing knowledge base. PMID- 19127234 TI - Staff perceptions of psychological care on a stroke rehabilitation unit. AB - This article describes the general principles of a psychological care model and the outcomes of a questionnaire survey with members of a multidisciplinary stroke rehabilitation team. Twenty-eight staff members' responses concerning their understanding and attitudes about the presented model were analysed. The responses revealed that rehabilitation staff had good knowledge about possible psychological reactions of patients following a stroke. They were supportive of the idea that it is important to understand and address patients' psychological needs to improve recovery. The survey revealed differing opinions regarding the way such a service could be implemented. Furthermore, the outcomes described personal requirements of rehabilitation staff in addressing the psychological care needs of patients. The results of the survey will have implications for staff training programmes and service developments. PMID- 19127235 TI - Mepitel: a non-adherent wound dressing with Safetac technology. AB - OBJECTIVE: Wound pain and tissue trauma are two main considerations of wound management, and appropriate dressing selection plays an important role in both. Traditional dressings may adhere to wounds resulting in significant pain and trauma to new tissue upon removal. The development of primary wound contact materials has provided a unique approach to solving this problem. This article aims to aid clinicians in identifying wound types on which Mepitel, a primary wound contact dressing with Safetac soft silicone adhesive technology, can be used by summarizing the published clinical literature relating to its use. METHOD: Searches of bibliographic databases and internet sites were supplemented with manual searches of journals of relevance to wound management for clinical data relating to the use of Mepitel. RESULTS: The literature search identified a number of articles, presenting data generated from randomized controlled trials, non-randomized controlled trials and case study evaluations of Mepitel on a wide range of wound types and skin injuries. CONCLUSION: The results of the clinical evaluations demonstrate that Mepitel is associated with atraumatic and virtually pain-free dressing changes. The dressing with Safetac can be used cost effectively in the treatment of a wide range of wound types and skin injuries. PMID- 19127236 TI - Review of 2008 and hopes for the future. PMID- 19127238 TI - Enaminones as building blocks in heterocyclic syntheses: reinvestigating the product structures of enaminones with malononitrile. A novel route to 6 substituted-3-oxo-2,3-dihydropyridazine-4-carboxylic acids. AB - The reported structures of reaction products of enaminones with malononitrile in ethanolic piperidine are revised. A novel route to 2,3-dihydropyridazine-4 carboxylic acids 4a-c via reactions of 2-cyano-5-(dimethylamino)-5-arylpenta-2,4 dienamides 8a-c with nitrous acid or with benzenediazonium chloride is reported. Compounds 8a-c are converted to 1,2-dihydropyridine-3-carboxylic acid and 1,2 dihydropyridine-3-carbonitrile derivatives upon reflux in EtOH/ HCl and in AcOH. PMID- 19127237 TI - Decreased oxygen transfer capacity of erythrocytes as a cause of 5-fluorouracil related ischemia. AB - Various mechanisms have been proposed to account for chemotherapy related ischemia, but none of them can explain the available clinical data. In order to explore the possibility that the decreased ability of erythrocytes to deliver oxygen to the heart could be responsible for cardiotoxicity, we have performed an ex vivo and in vivo study of the effects of cisplatin/5-FU on erythrocytes, using a variety of biophysical techniques. Combining EPR and microscopy it was concluded that both cardiotoxic 5-FU and non-cardiotoxic cisplatin have similar effects on the erythrocyte membrane, thus eliminating those changes as a potential source of cardiotoxicity. On the contrary, 31P-NMR and polarography showed that the effects of these cytostatics on the intracellular milieu differ significantly. 5-FU provoked a pronounced decrease of the O2 level in blood and affected the metabolism of phosphate compounds, while cisplatin had no such effects. When combined these two drugs showed synergistic effects, which matches the higher frequency of cardiotoxicity of the combination relative to the sole application of 5-FU. Preliminary results acquired on blood of patients receiving cisplatin/5-FU therapy verified observations obtained ex vivo. These results open a possibility of applying NMR in preclinical trials of new drugs in order to predict their ischemic potential. PMID- 19127239 TI - Synthesis of 5-substituted 3-amino-1H-pyrazole-4-carbonitriles as precursors for microwave assisted regiospecific syntheses of pyrazolo[1,5-a]pyrimidines. AB - A simple route to 3-oxoalkanonitrile 5, aprecursor of the title compounds is described. Reaction of enaminones 2 with hydroxylamine hydrochloride in ethanol yielded aldoximes 3 that were converted readily into 5 in basic medium. This method has been successfully applied with a number of substrates and resulted in excellent yields of the products. Reacting 5 with trichloroacetonitrile afforded 3-amino-2-aroyl-4,4,4-trichloro-2-butenenitriles 6 that condensed with hydrazines to yield 3-amino-1H-pyrazole-4-carbonitrilederivatives 8. Substituted pyrazolo[1,5-a]pyridmidines have been prepared with regioselective condensation reactions of 8 with nonsymmetrical dielectrophiles. The structures of compounds obtained were deduced based on (1)H-NMR, (1)H-(15)N HMBC- measurements. PMID- 19127240 TI - Total synthesis and the biological activities of (+/-)-norannuradhapurine. AB - The structure previously assigned to the phenolic noraporphine alkaloid, (-) norannuradhapurine has been confirmed by a total synthesis of the racemic alkaloid in which the key step involved the formation of the C ring by a radical initiated cyclization. although inactive against Staphylococcus aureus ATCC25932, Escherichia coli ATCC10536 and Candida albicans ATCC90028, (+/-) norannuradhapurine inhibits the production of NO, PGE(2), TNF-alpha, IL-1beta and IL-6 and the expression of iNOS and COX-2 in RAW 264.7 macrophages stimulated with LPS in vitro. PMID- 19127241 TI - The inhibitory action of kohamaic acid A derivatives on mammalian DNA polymerase beta. AB - We previously isolated a novel natural product, designated kohamaic acid A (KA-A, compound 1), as an inhibitor of the first cleavage of fertilized sea urchin eggs, and found that this compound could selectively inhibit the activities of mammalian DNA polymerases (pols). In this paper, we investigated the structure and bioactivity of KA-A and its chemically synthesized 11 derivatives (i.e., compounds 2-12), including KA-A - fatty acid conjugates. The pol inhibitory activity of compound 11 [(1S*,4aS*,8aS*)-17-(1,4,4a,5,6,7,8,8a-octahydro-2,5,5,8a tetramethyl-naphthalen-1-yl)heptadecanoic acid] was the strongest among the synthesized compounds, and the range of IC(50) values for mammalian pols was 3.22 to 8.76 microM; therefore, the length of the fatty acid side chain group of KA-A is important for pol inhibition. KA-A derivatives could prevent human cancer cell (promyelocytic leukemia cell line, HL-60) growth with the same tendency as the inhibition of mammalian pols. Since pol beta is the smallest molecule, we used it to analyze the biochemical relationship with KA-A derivatives. From computer modeling analysis (i.e., docking simulation analysis), these compounds bound selectively to four amino acid residues (Leu11, Lys35, His51 and Thr79) of the N terminal 8-kDa domain of pol beta, and the binding energy between compound 11 and pol beta was largest in the synthesized compounds. The relationship between the three-dimensional molecular structures of KA-A-related compounds and these inhibitory activities is discussed. PMID- 19127242 TI - Differential cytotoxicity of MEX: a component of Neem oil whose action is exerted at the cell membrane level. AB - Neem oil is obtained from the seeds of the tree Azadirachta indica. Its chemical composition is very complex, being rich in terpenoids and limonoids, as well as volatile sulphur modified compounds. This work focused on the evaluation of a component of the whole Neem oil obtained by methanolic extraction and defined as MEX. Cytotoxicity was assessed on two different cell populations: a stabilized murine fibroblast line (3T6) and a tumor cell line (HeLa). The data presented here suggest a differential sensitivity of these two populations, the tumor line exhibiting a significantly higher sensitivity to MEX. The data strongly suggest that its toxic target is the cell membrane. In addition the results presented here imply that MEX may contain one or more agents that could find a potential use in anti-proliferative therapy. PMID- 19127243 TI - Antioxidant activities of polyphenols extracted from Perilla frutescens varieties. AB - Various cultivars of Perilla frutescens (L.) (var. crispa and var. frutescens) Britt. were harvested in China and Japan. They were easily differentiated on the basis of their foliage color, that varied from red to green. Water extracts of dried plants were investigated for their antioxidant activity (AA) and their polyphenolic compounds compared. Among them, cinnamic acid derivatives (coumaroyl tartaric acid, caffeic acid and rosmarinic acid), flavonoids (apigenin 7-O caffeoylglucoside, scutellarein 7-O-diglucuronide, luteolin 7-O-diglucuronide, apigenin 7-O-diglucuronide, luteolin 7-O-glucuronide, and scutellarein 7-O glucuronide) and anthocyanins (mainly cis-shisonin, shisonin, malonylshisonin and cyanidin 3-O-(E)-caffeoylglucoside-5-O-malonylglucoside) were quantified. AA assays are based on the inhibition of the free radical 2,2-diphenyl-1 picrylhydrazyl (DPPH). The DPPH radical scavenging activity was calculated as Trolox [(+/-)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid] equivalent antioxidant capacity (TEAC). The mean amount of total phenolics of the water extracts (4-29 micromol/100 mL) and the TEAC value calculated (23-167 micromol TE/100 mL) confirmed the high antioxidant activity of these leaf water extracts. These results were highly correlated within some o-dihydroxylated polyphenolic compounds and AA. PMID- 19127244 TI - Selenium status and cardiovascular risk profile in healthy adult Saudi males. AB - The purpose of this research was to investigate the relationship between selenium levels, thyroid function and other coronary risk factors in 140 Saudi subjects without overt coronary heart disease stratified by age. Demographic data and serum fasting lipid profile, glucose, thyroid function tests, selenium status and dietary intake was assessed. The relationships between selenium status, thyroid function and cardiovascular risk factors were assessed by univariate and multivariate analysis. The results showed that thyroid hormone levels did not differ with age. Erythrocyte glutathione peroxidase (GPx) levels were significantly higher in the youngest vs. oldest tertile (p<0.0001). Selenium and iodine intake did not differ significantly with age tertile, but the average intake for the population sample was below the estimated average requirements for both elements. Serum lipoprotein (a) concentrations correlated with selenium (r = 0.417, p<0.0001) and TSH (r = 0.172, p<0.05). After adjustment for confounding variables; serum fT(4) and erythrocytes GPx remained significant determinants of serum TSH levels, whilst serum selenium and TSH were determinants of serum fT(4) levels. Serum Lp(a), a coronary risk factor, was strongly related to measures of selenium status. A significant relationship between measures of selenium status and thyroid function was found. Serum Lp(a) a known risk factor for cardiovascular disease was also related to selenium status in our population. PMID- 19127245 TI - Synthesis and phytogrowth properties of oxabicyclic analogues related to helminthosporin. AB - This investigation describes the synthesis and biological evaluation of a series of oxabicyclic analogues related to the helminthosporins. Four oxabicycles were prepared by [4+3] cycloaddition of an oxyallyl carbocation, generated in situ from 2,4-dibromopentan-3-one, with selected furans. Functional group manipulations of the oxabicyclic architecture generated nine further derivatives. The phytotoxic properties of these oxabicycles were evaluated as their ability to interfere with the growth of Sorghum bicolor and Cucumis sativus seedlings. In both species, the most active compounds were oxabicycles possessing a carbonyl group conjugated with a double bond. PMID- 19127246 TI - Coordination modes of a schiff base pentadentate derivative of 4-aminoantipyrine with cobalt(II), nickel(II) and copper(II) metal ions: synthesis, spectroscopic and antimicrobial studies. AB - Transition metal complexes of Co(II), Ni(II) and Cu(II) metal ions with general stoichiometry [M(L)X]X and [M(L)SO(4)], where M = Co(II), Ni(II) and Cu(II), L = 3,3'-thiodipropionic acid bis(4-amino-5-ethylimino-2,3-dimethyl-1-phenyl-3 pyrazoline) and X = NO(3)(-), Cl(-) and OAc(-), have been synthesized and structurally characterized by elemental analyses, molar conductance measurements, magnetic susceptibility measurements and spectral techniques like IR, UV and EPR. The nickel(II) complexes were found to have octahedral geometry, whereas cobalt(II) and copper(II) complexes were of tetragonal geometry. The covalency factor (beta) and orbital reduction factor (k) suggest the covalent nature of the complexes. The ligand and its complexes have been screened for their antifungal and antibacterial activities against three fungi, i.e. Alternaria brassicae, Aspergillus niger and Fusarium oxysporum and two bacteria, i.e. Xanthomonas compestris and Pseudomonas aeruginosa. PMID- 19127247 TI - Pimarane-type diterpenes: antimicrobial activity against oral pathogens. AB - Seven pimarane type-diterpenes re-isolated from Viguiera arenaria Baker and two semi-synthetic pimarane derivatives were evaluated in vitro against the following main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei. The compounds ent-pimara-8(14),15-dien-19-oic acid (PA); ent-8(14),15-pimaradien 3beta-ol; ent-15-pimarene-8beta,19-diol; ent-8(14),15-pimaradien-3beta-acetoxy and the sodium salt derivative of PA were the most active compounds, displaying MIC values ranging from 2 to 8 microg mL(-1). Thus, this class of compounds seems promising as a class of new effective anticariogenic agents. Furthermore, our results also allow us to conclude that minor structural differences among these diterpenes significantly influence their antimicrobial activity, bringing new perspectives to the discovery of new natural compounds that could be employed in the development of oral care products. PMID- 19127248 TI - Phenolic and theobromine contents of commercial dark, milk and white chocolates on the Malaysian market. AB - Chocolate contains a wide range of antioxidants that includes soluble phenolic compounds (phenolic acids, catechin, epicatechin, and proanthocyanidins), insoluble polymeric phenolics and methylxanthines. The objective of this study was to determine phenolic and theobromine contents in dark (DC), milk (MC), and white (WC) chocolates commonly found in the Malaysian marketplace. Total phenolic and flavonoids were determined by means of a spectrometric assay, while catechin, epicatechin and theobromine were quantified using a reverse-phase HPLC method. Dark chocolates exhibited the highest phenolics and flavonoids contents, followed by milk and white chocolates. Catechin and epicatechin were major flavonoids detected in dark chocolates. Theobromine was detected in dark and milk chocolates, but not in white chocolates. A high correlation (r= 0.93) between total phenolic and flavonoid contents, indicating that the major phenolic compounds in dark chocolates belong to the flavonoid class. When nutrition and health promotion are of concern, dark chocolates would be recommended over milk and white chocolates owing to their higher contents of antioxidant phenolic compounds. PMID- 19127249 TI - The use of coumarins as environmentally-sensitive fluorescent probes of heterogeneous inclusion systems. AB - Coumarins, as a family of molecules, exhibit a wide range of fluorescence emission properties. In many cases, this fluorescence is extremely sensitive to the local environment of the molecule, especially the local polarity and microviscosity. In addition, coumarins show a wide range of size, shape, and hydrophobicity. These properties make them especially useful as fluorescent probes of heterogeneous environments, such as supramolecular host cavities, micelles, polymers and solids. This article will review the use of coumarins to probe such heterogeneous systems using fluorescence spectroscopy. PMID- 19127250 TI - Enhanced factor VIII heavy chain for gene therapy of hemophilia A. AB - Hemophilia A gene therapy using recombinant adenovirus-associated virus (AAV) vectors has been hampered by the size of the factor VIII (FVIII) cDNA. Previously, splitting the FVIII coding sequence into a heavy-chain (HC) fragment and a light-chain (LC) fragment for dual recombinant AAV vector delivery has been successfully explored. However, the main disadvantage of this approach is a "chain imbalance" problem in which LC secretion is approximately 1-2 logs higher than that of HC, and therefore, the majority of protein synthesized is nonfunctional. To improve HC secretion, we constructed alternate FVIII HCs based on our observation that LC facilitates HC secretion. To our surprise, most of the new HC molecules exhibited enhanced expression over the traditional HC molecule (HC(745)). The optimized HC mutein, HC(HL), including additional acidic-region-3 (ar3) sequences, exhibited three- to fivefold higher activity in both enzyme linked immunosorbent assay (ELISA) and activated partial thromboplastin time (aPTT) assay in in vitro testing. Further characterization suggested ar3 sequences increased HC secretion, rather than promoting HC synthesis. Intravenous delivery of AAV8-HC(HL)+AAV8-LC or AAV8-HC(745)+AAV8-LC achieved phenotypic correction in hemophilia A mice. Mice receiving AAV8-HC(HL)+AAV8-LC achieved three- to fourfold higher HC expression than AAV8-HC(745)+AAV8-LC, consistent with the FVIII functional assays. HC(HL) should be substituted for HC(745) in a dual AAV vector strategy due to its enhanced expression. PMID- 19127251 TI - Human neural stem cells can target and deliver therapeutic genes to breast cancer brain metastases. AB - The tumor-tropic properties of neural stem cells (NSCs) led to the development of a novel strategy for delivering therapeutic genes to tumors in the brain. To apply this strategy to the treatment of brain metastases, we made a human NSC line expressing cytosine deaminase (F3.CD), which converts 5-fluorocytosine (5 FC) into 5-fluorouracil, an anticancer agent. In vitro, the F3.CD cells significantly inhibited the growth of tumor cell lines in the presence of the prodrug 5-FC. In vivo, MDA-MB-435 human breast cancer cells were implanted into the brain of immune-deficient mouse stereotactically, and F3.CD cells were injected into the contralateral hemisphere followed by systemic 5-FC administration. The F3.CD cells migrated selectively into the brain metastases located in the opposite hemisphere and resulted in significantly reduced volumes. The F3.CD and 5-FC treatment also decreased both tumor volume and number of tumor mass significantly, when immune-deficient mouse had MDA-MB-435 cells injected into the internal carotid artery and F3.CD cells were transplanted into the contralateral brain hemisphere stereotactically. Taken together, brain transplantation of human NSCs, encoding the suicide enzyme CD, combined with systemic administration of the prodrug 5-FC, is an effective treatment regimen for brain metastases of tumors. PMID- 19127253 TI - Breast cancer incidence, stage, treatment and survival in ethnic groups in South East England. AB - Studies from the US have shown variations in breast cancer incidence, stage distribution, treatment and survival between ethnic groups. Data on 35 631 women diagnosed with breast cancer in South East England between 1998 and 2003 with self-assigned ethnicity information available were analysed. Results are reported for White, Indian, Pakistani, Bangladeshi, Black Caribbean, Black African and Chinese women. Age-standardised breast cancer incidence rate ratios, patterns of stage of disease at diagnosis, treatment, overall and breast cancer-specific survival were examined. All ethnic groups studied had lower age-standardised breast cancer incidence rates than White women, with Bangladeshi women having the lowest rate ratio (0.23, 95% CI: 0.20-0.26). White women were the most likely to have a stage recorded at diagnosis (adjusted proportion 75%), and least likely to be diagnosed with metastatic disease (7%). Black African women were the least likely to have a record of cancer surgery (63%) or hormone therapy (32%), and most likely to receive chemotherapy (38%). After fully adjusting for age, socioeconomic deprivation, stage of disease and treatment received, there was no significant variation in breast cancer-specific survival. However, Black African women had significantly worse overall survival (hazard ratio 1.24, P=0.025). These findings suggest that a strategy of earlier detection should be pursued in Black and South Asian women. PMID- 19127252 TI - Cancer-induced expansion and activation of CD11b+ Gr-1+ cells predispose mice to adenoviral-triggered anaphylactoid-type reactions. AB - Intravascular delivery (1.5 x 10(9) particles and higher) of recombinant adenovirus (rAd) induces myeloid cell mediated, self-limiting hemodynamic responses in normal mice. However, we observed anaphylactoid-type reactions and exacerbated hemodynamic events following rAd injection in mice bearing malignant 4T1 mammary carcinoma. Because 4T1 tumors induce significant CD11b(+)Gr-1(+) myeloid cell expansion and activation, we set to determine whether this causes rAd-induced exaggerated responses. When treated with a single intravenous dose (1 x 10(10) particles) of rAd, mice implanted with 4T1 carcinoma succumbed due to the anaphylactoid-type reactions. In contrast, normal mice and mice implanted with a related mammary carcinoma (66cl4) that does not induce CD11b(+)Gr-1(+) cell expansion, showed minimal responses. Depletion of phagocytic CD11b(+)Gr-1(+) cells prior to rAd delivery protected 4T1 tumor-bearing animals, whereas passive transfer of CD11b(+)Gr-1(+) cells from 4T1 tumor-bearing animals was sufficient to convey susceptibility to anaphylactoid-type reactions in normal animals. We further show that there is upregulation of nitric oxide and leukotriene signaling pathways in the 4T1 tumor-induced CD11b(+)Gr-1(+) myeloid cells and that pretreating mice with inhibitors of nitric oxide synthetase and leukotrienes can attenuate the anaphylactoid-type reactions. These data show that malignant tumor growth can alter CD11b(+)Gr-1(+) myeloid cells, rendering hosts susceptible to anaphylactoid-type reactions upon intravascular treatment with rAd. PMID- 19127254 TI - Normal stem cells in cancer prone epithelial tissues. AB - The concept of a cancer stem cell is not a new one, being first suggested over 100 years ago. Over recent years the concept has enjoyed renewed enthusiasm, partly because of our growing understanding of the nature of somatic stem cells, but also because of a growing realisation that the development of strategies that target cancer stem cells may offer considerable advantages over conventional approaches. However, despite this renewed enthusiasm the existence of cancer stem cells remains controversial in many tumour types and any potential relationship to the normal stem cell pool remains poorly defined. This review summarises key elements of our understanding of the normal stem cell populations within animal models of the predominant cancer prone epithelial tissues, and further investigates the potential links between these populations and putative cancer stem cells. PMID- 19127256 TI - Safety and pharmacokinetics of paclitaxel and the oral mTOR inhibitor everolimus in advanced solid tumours. AB - Everolimus displays antiproliferative effects on cancer cells, yields antiangiogenic activity in established tumours, and shows synergistic activity with paclitaxel in preclinical models. This study assessed the safety and the pharmacokinetic interactions of everolimus and paclitaxel in patients with advanced malignancies. Everolimus was dose escalated from 15 to 30 mg and administered with paclitaxel 80 mg m(-2) on days 1, 8, and 15 every 28 days. Safety was assessed weekly, and dose-limiting toxicity (DLT) was evaluated in cycle 1. A total of 16 patients (median age 54.5 years, range 33-69) were entered; 11 had prior taxane therapy for breast (n=5), ovarian (n=3), and vaginal cancer (n=1) or angiosarcoma (n=2). Grade 3 neutropenia in six patients met the criteria for DLT in two patients receiving everolimus 30 mg weekly. Other drug related grade 3 toxicities were leucopenia, anaemia, thrombocytopenia, stomatitis, asthenia, and increased liver enzymes. Tumour stabilisation reported in 11 patients exceeded 6 months in 2 patients with breast cancer. Everolimus showed an acceptable safety profile at the dose of 30 mg when combined with weekly paclitaxel 80 mg m(-2), warranting further clinical investigation. PMID- 19127255 TI - Validating genetic risk associations for ovarian cancer through the international Ovarian Cancer Association Consortium. AB - The search for genetic variants associated with ovarian cancer risk has focused on pathways including sex steroid hormones, DNA repair, and cell cycle control. The Ovarian Cancer Association Consortium (OCAC) identified 10 single-nucleotide polymorphisms (SNPs) in genes in these pathways, which had been genotyped by Consortium members and a pooled analysis of these data was conducted. Three of the 10 SNPs showed evidence of an association with ovarian cancer at P< or =0.10 in a log-additive model: rs2740574 in CYP3A4 (P=0.011), rs1805386 in LIG4 (P=0.007), and rs3218536 in XRCC2 (P=0.095). Additional genotyping in other OCAC studies was undertaken and only the variant in CYP3A4, rs2740574, continued to show an association in the replication data among homozygous carriers: OR(homozygous(hom))=2.50 (95% CI 0.54-11.57, P=0.24) with 1406 cases and 2827 controls. Overall, in the combined data the odds ratio was 2.81 among carriers of two copies of the minor allele (95% CI 1.20-6.56, P=0.017, p(het) across studies=0.42) with 1969 cases and 3491 controls. There was no association among heterozygous carriers. CYP3A4 encodes a key enzyme in oestrogen metabolism and our finding between rs2740574 and risk of ovarian cancer suggests that this pathway may be involved in ovarian carcinogenesis. Additional follow-up is warranted. PMID- 19127258 TI - A recurrent truncating germline mutation in the BRIP1/FANCJ gene and susceptibility to prostate cancer. AB - Although prostate cancer (PrCa) is one of the most common cancers in men in Western countries, little is known about the inherited factors that influence PrCa risk. On the basis of the fact that BRIP1/FANCJ interacts with BRCA1 and functions as a regulator of DNA double-strand break repair pathways, and that germline mutations within the BRIP1/FANCJ gene predispose to breast cancer, we chose this gene as a candidate for mutation screening in familial and young-onset PrCa cases. We identified a truncating mutation, R798X, in the BRIP1/FANCJ gene in 4 out of 2714 UK PrCa cases enriched for familial (2 out of 641; 0.3%) and young-onset cases (2 out of 2073; 0.1%). On screening 2045 controls from the UK population, we found one R798X sequence alteration (0.05%; odds ratio 2.4 (95% CI 0.25-23.4)). In addition, using our data from a genome-wide association study, we analysed 25 SNPs in the genomic region of the BRIP1/FANCJ gene. Two SNPs showed evidence of association with familial and young-onset PrCa (rs6504074; P(trend)=0.04 and rs8076727; P(trend)=0.01). These results suggest that truncating mutations in BRIP1/FANCJ might confer an increased risk of PrCa and common SNPs might also contribute to the alteration of risk, but larger case control series will be required to confirm or refute this association. PMID- 19127257 TI - Temozolomide- and fotemustine-induced apoptosis in human malignant melanoma cells: response related to MGMT, MMR, DSBs, and p53. AB - Malignant melanomas are highly resistant to chemotherapy. First-line chemotherapeutics used in melanoma therapy are the methylating agents dacarbazine (DTIC) and temozolomide (TMZ) and the chloroethylating agents BCNU and fotemustine. Here, we determined the mode of cell death in 11 melanoma cell lines upon exposure to TMZ and fotemustine. We show for the first time that TMZ induces apoptosis in melanoma cells, using therapeutic doses. For both TMZ and fotemustine apoptosis is the dominant mode of cell death. The contribution of necrosis to total cell death varied between 10 and 40%. The O(6)-methylguanine DNA methyltransferase (MGMT) activity in the cell lines was between 0 and 1100 fmol mg(-1) protein, and there was a correlation between MGMT activity and the level of resistance to TMZ and fotemustine. MGMT inactivation by O(6) benzylguanine sensitized all melanoma cell lines expressing MGMT to TMZ and fotemustine-induced apoptosis, and MGMT transfection attenuated the apoptotic response. This supports that O(6)-alkylguanines are critical lesions involved in the initiation of programmed melanoma cell death. One of the cell lines (MZ7), derived from a patient subjected to DTIC therapy, exhibited a high level of resistance to TMZ without expressing MGMT. This was related to an impaired expression of MSH2 and MSH6. The cells were not cross-resistant to fotemustine. Although these data indicate that methylating drug resistance of melanoma cells can be acquired by down-regulation of mismatch repair, a correlation between MSH2 and MSH6 expression in the different lines and TMZ sensitivity was not found. Apoptosis in melanoma cells induced by TMZ and fotemustine was accompanied by double-strand break (DSB) formation (as determined by H2AX phosphorylation) and caspase-3 and -7 activation as well as PARP cleavage. For TMZ, DSBs correlated significantly with the apoptotic response, whereas for fotemustine a correlation was not found. Melanoma lines expressing p53 wild-type were more resistant to TMZ and fotemustine than p53 mutant melanoma lines, which is in marked contrast to previous data reported for glioma cells treated with TMZ. Overall, the findings are in line with the model that in melanoma cells TMZ-induced O(6)-methylguanine triggers the apoptotic (and necrotic) pathway through DSBs, whereas for chloroethylating agents apoptosis is triggered in a more complex manner. PMID- 19127259 TI - Phase II study and biomarker analysis of cetuximab combined with modified FOLFOX6 in advanced gastric cancer. AB - This prospective study was conducted with the Korean Cancer Study Group to evaluate the efficacy and safety of cetuximab combined with modified FOLFOX6 (mFOLFOX6) as first-line treatment in recurrent or metastatic gastric cancer and to identify potential predictive biomarkers. Patients received cetuximab 400 mg m(-2) at week 1 and 250 mg m(-2) weekly thereafter until disease progression. Oxaliplatin (100 mg m(-2)) and leucovorin (100 mg m(-2)) were administered as a 2 h infusion followed by a 46-h continuous infusion of 5-fluorouracil (2400 mg m( 2)) every 2 weeks for a maximum of 12 cycles. Biomarkers potentially associated with efficacy were analysed. Among 38 evaluable patients, confirmed response rate (RR) was 50.0% (95% CI 34.1-65.9). Median time-to-progression (TTP) was 5.5 months (95% CI 4.5-6.5) and overall survival (OS) 9.9 months. Eleven patients having tumour EGFR expression by immunohistochemistry with low serum EGF and TGF alpha levels showed a 100% RR compared to 37.0% in the remaining 27 patients (P<0.001). Moreover, ligand level increased when disease progressed in seven out of eight patients with EGFR expression and low baseline ligand level. No patient exhibited EGFR amplification or K-ras mutations. Gastric cancer patients with EGFR expression and low ligand levels had better outcomes with cetuximab/mFOLFOX6 treatment. PMID- 19127260 TI - Adjuvant 5-fluorouracil and folinic acid vs observation for pancreatic cancer: composite data from the ESPAC-1 and -3(v1) trials. AB - The ESPAC-1, ESPAC-1 plus, and early ESPAC-3(v1) results (458 randomized patients; 364 deaths) were used to estimate the effectiveness of adjuvant 5FU/FA vs resection alone for pancreatic cancer using meta-analysis. The pooled hazard ratio of 0.70 (95% CI=0.55-0.88) P=0.003, and the median survival of 23.2 (95% CI=20.1-26.5) months with 5FU/FA vs 16.8 (95% CI=14.3-19.2) months with resection alone supports the use of adjuvant 5FU/FA in pancreatic cancer. PMID- 19127261 TI - A phase III trial of topotecan and whole brain radiation therapy for patients with CNS-metastases due to lung cancer. AB - Brain metastases represent an important cause of morbidity in patients with lung cancer and are associated with a mean survival of less than 6 months. Thus, new regimens improving the outcome of these patients are urgently needed. On the basis of promising data raised in a phase I/II trial, we initiated an open, randomised, prospective, multicentric phase III trial, comparing whole brain radiation therapy (WBRT; 20 x 2 Gy) alone with WBRT+topotecan (RCT; 0.4 mg m(-2) day(-1) x 20). A total of 320 patients with CNS-metastases due to SCLC or NSCLC were projected. The primary end point was overall survival, whereas second end points were local response and progression-free survival. However, until the cutoff date of study completion (i.e., a study duration of 34 months), only a total of 96 (RCT:47, WBRT:49) patients had been recruited, and so an analysis was performed at that time point. Although the numbers of grade 3/4 non haematological toxicities (besides alopecia 115 (RCT/WBRT: 55 out of 60) were evenly distributed, the 25 haematological events occurred mainly in the combined treatment arm (24 out of 1). Local response, evaluated 2 weeks after treatment, was assessable in 44 (RCT/WBRT: 23 out of 21) patients, showing CR in eight (3 out of 5), PR in 17 (11 out of 6), SD in 14 (8 out of 6) and PD in five (1 out of 4) patients (all differences n.s.). Neither OAS (RCT/WBRT: median (days)): 87 out of 95, range 3-752/4-433; HR 1.32; 95% CI (0.83; 2.10)) nor PFS (median (days)): 71 out of 66, range, 3-399/4-228; HR 1.28, 95% CI (0.73; 2.43) differed significantly. On the basis of these results and the slow recruitment, a continuation of the study did not seem reasonable. The available data show no significant advantage for concurrent radiochemotherapy for patients with lung cancer; however, the recruited number of patients is too low to exhibit a small advantage of combined treatment. PMID- 19127262 TI - Different cervical cancer screening approaches in a Chinese multicentre study. AB - To evaluate alternative cervical cancer screening methods, digital colposcopy and collection of cervical exfoliated cells for liquid-based cytology (LBC) and hybrid capture 2 (HC2) testing were performed among 2562 women aged 15-59 years in three study sites in the People's Republic of China (rural Shanxi province, Shenyang city in Liaoning province and Shenzhen city in Guangdong province). Visual inspection with acetic acid (VIA) was also evaluated independently from colposcopy. A total of 74 cases of histologically confirmed cervical intraepithelial neoplasia grade 2 or worse (CIN2+) were identified, and 16 CIN2+ cases were imputed among unbiopsied women to correct for verification bias. Corrected sensitivity for CIN2+ was 37% for VIA, 54% for colposcopy, 87% for LBC with a threshold of atypical cells of undetermined significance (LBC>or=ASCUS), 90% for HC2, 84% for LBC using HC2 to triage ASCUS and 96% for positivity to LBC>or=ASCUS or HC2. For VIA, sensitivity was much lower among women >or=40 years (12%) than those aged or=ASCUS or HC2, up to 94% for LBC using HC2 to triage ASCUS. In conclusion, LBC, HC2 and their combinations performed well, whereas VIA missed a majority of CIN2+, particularly in older women. Digital colposcopy performed better than VIA, but still missed nearly half of CIN2+ in this study. PMID- 19127264 TI - Recognition of skin malignancy by general practitioners: observational study using data from a population-based randomised controlled trial. AB - Skin malignancy is an important cause of mortality in the United Kingdom and is rising in incidence every year. Most skin cancer presents in primary care, and an important determinant of outcome is initial recognition and management of the lesion. Here we present an observational study of interobserver agreement using data from a population-based randomised controlled trial of minor surgery. Trial participants comprised patients presenting in primary care and needing minor surgery in whom recruiting doctors felt to be able to offer treatment themselves or to be able to refer to a colleague in primary care. They are thus relatively unselected. The skin procedures undertaken in the randomised controlled trial generated 491 lesions with a traceable histology report: 36 lesions (7%) from 33 individuals were malignant or pre-malignant. Chance-corrected agreement (kappa) between general practitioner (GP) diagnosis of malignancy and histology was 0.45 (0.36-0.54) for lesions and 0.41 (0.32-0.51) for individuals affected with malignancy. Sensitivity of GPs for the detection of malignant lesions was 66.7% (95% confidence interval (CI), 50.3-79.8) for lesions and 63.6% (95% CI, 46.7 77.8) for individuals affected with malignancy. The safety of patients is of paramount importance and it is unsafe to leave the diagnosis and treatment of potential skin malignancy in the hands of doctors who have limited training and experience. However, the capacity to undertake all of the minor surgical demand works demanded in hospitals does not exist. If the capacity to undertake it is present in primary care, then the increased costs associated with enhanced training for general medical practitioners (GPs) must be borne. PMID- 19127263 TI - Promoter CpG methylation in cancer cells contributes to the regulation of MUC4. AB - Mucin 4 (MUC4) is a high molecular weight transmembrane mucin that is overexpressed in many carcinomas and is a risk factor associated with a poor prognosis. In this study, we show that the DNA methylation pattern is intimately correlated with MUC4 expression in breast, lung, pancreas and colon cancer cell lines. We mapped the DNA methylation status of 94 CpG sites from -3622 to +29 using MassARRAY analysis that utilises base-specific cleavage of nucleic acids. MUC4-negative cancer cell lines and those with low MUC4 expression (eg, A427) were highly methylated near the transcriptional start site, whereas MUC4-positive cell lines (eg, NCI-H292) had low methylation levels. Moreover, 5-aza-2' deoxycytidine and trichostatin A treatment of MUC4-negative cells or those with low MUC4 expression caused elevation of MUC4 mRNA. Our results suggest that DNA methylation in the 5' flanking region play an important role in MUC4 gene expression in carcinomas of various organs. An understanding of epigenetic changes in MUC4 may contribute to the diagnosis of carcinogenic risk and prediction of outcome in patients with cancer. PMID- 19127265 TI - A phase I-II study of the histone deacetylase inhibitor valproic acid plus chemoimmunotherapy in patients with advanced melanoma. AB - We explored in a phase I/II clinical trial the combination of valproic acid (VPA), a clinically available histone deacetylase inhibitor, with standard chemoimmunotherapy in patients with advanced melanoma, to evaluate its clinical activity, to correlate the clinical response with the biological activity of VPA and to assess toxicity. Patients were treated initially with VPA alone for 6 weeks. The inhibition of the target in non-tumour peripheral blood cells (taken as a potential surrogate marker) was measured periodically, and valproate dosing adjusted with the attempt to reach a measurable inhibition. After the treatment with valproate alone, dacarbazine plus interferon-alpha was started in combination with valproate. Twenty-nine eligible patients started taking valproate and 18 received chemoimmunotherapy and are assessable for response. We observed one complete response, two partial remissions and three disease stabilisations lasting longer than 24 weeks. With the higher valproate dosages needed to reach a measurable inhibition of the target, we observed an increase of side effects in those patients who received chemoimmunotherapy. The combination of VPA and chemoimmunotherapy did not produce results overtly superior to standard therapy in patients with advanced melanoma and toxicity was not negligible, casting some doubts on the clinical use of VPA in this setting (at least in the administration schedule adopted). PMID- 19127266 TI - The influence of systemic inflammation, dietary intake and stage of disease on rate of weight loss in patients with gastro-oesophageal cancer. AB - Although weight loss is often a dominant symptom in patients with upper gastrointestinal malignancy, there is a lack of objective evidence describing changes in nutritional status and potential associations between weight loss, food intake, markers of systemic inflammation and stage of disease in such patients. Two hundred and twenty patients diagnosed with gastric/oesophageal cancer were studied. Patients underwent nutritional assessment consisting of calculation of body mass index, measurement of weight loss, dysphagia scoring and estimation of dietary intake. Serum acute-phase protein concentrations were determined by enzyme-linked immunosorbent assay. In all, 182 (83%) patients had lost weight at diagnosis (median loss, 7% body weight). Weight loss was associated with poor performance status, advanced disease stage, dysphagia, reduced dietary intake and elevated serum C-reactive protein (CRP) concentrations. Multiple regression identified dietary intake (estimate of effect, 38%), serum CRP concentrations (estimate of effect, 34%) and stage of disease (estimate of effect, 28%) as independent variables in determining degree of weight loss. Mechanisms other than reduced dietary intake or mechanical obstruction by the tumour appear to be involved in the nutritional decline in patients with gastro-oesophageal malignancy. Recognition that systemic inflammation plays a role in nutritional depletion may inform the development of appropriate therapeutic strategies to ameliorate weight loss, making patients more tolerant of cancer-modifying treatments such as chemotherapy. PMID- 19127267 TI - ATP7A is a novel target of retinoic acid receptor beta2 in neuroblastoma cells. AB - Increased retinoic acid receptor beta (RARbeta(2)) gene expression is a hallmark of cancer cell responsiveness to retinoid anticancer effects. Moreover, low basal or induced RARbeta(2) expression is a common feature of many human cancers, suggesting that RARbeta(2) may act as a tumour suppressor gene in the absence of supplemented retinoid. We have previously shown that low RARbeta(2) expression is a feature of advanced neuroblastoma. Here, we demonstrate that the ABC domain of the RARbeta(2) protein alone was sufficient for the growth inhibitory effects of RARbeta(2) on neuroblastoma cells. ATP7A, the copper efflux pump, is a retinoid responsive gene, was upregulated by ectopic overexpression of RARbeta(2). The ectopic overexpression of the RARbeta(2) ABC domain was sufficient to induce ATP7A expression, whereas, RARbeta(2) siRNA blocked the induction of ATP7A expression in retinoid-treated neuroblastoma cells. Forced downregulation of ATP7A reduced copper efflux and increased viability of retinoid-treated neuroblastoma cells. Copper supplementation enhanced cell growth and reduced retinoid-responsiveness, whereas copper chelation reduced the viability and proliferative capacity. Taken together, our data demonstrates ATP7A expression is regulated by retinoic acid receptor beta and it has effects on intracellular copper levels, revealing a link between the anticancer action of retinoids and copper metabolism. PMID- 19127269 TI - Second primary cancers in patients with skin cancer: a population-based study in Northern Ireland. AB - Among all 14,500 incident cases of basal cell carcinoma (BCC), 6405 squamous cell carcinomas (SCC) and 1839 melanomas reported to the Northern Ireland Cancer Registry between 1993 and 2002, compared with the general population, risk of new primaries after BCC or SCC was increased by 9 and 57%, respectively. The subsequent risk of cancer, overall, was more than double after melanoma. PMID- 19127268 TI - LLL-3 inhibits STAT3 activity, suppresses glioblastoma cell growth and prolongs survival in a mouse glioblastoma model. AB - Persistent activation of the signal transducer and activator of transcription 3 (STAT3) signalling has been linked to oncogenesis and the development of chemotherapy resistance in glioblastoma and other cancers. Inhibition of the STAT3 pathway thus represents an attractive therapeutic approach for cancer. In this study, we investigated the inhibitory effects of a small molecule compound known as LLL-3, which is a structural analogue of the earlier reported STAT3 inhibitor, STA-21, on the cell viability of human glioblastoma cells, U87, U373, and U251 expressing constitutively activated STAT3. We also investigated the inhibitory effects of LLL-3 on U87 glioblastoma cell growth in a mouse tumour model as well as the impact it had on the survival time of the treated mice. We observed that LLL-3 inhibited STAT3-dependent transcriptional and DNA binding activities. LLL-3 also inhibited viability of U87, U373, and U251 glioblastoma cells as well as induced apoptosis of these glioblastoma cell lines as evidenced by increased poly (ADP-ribose) polymerase (PARP) and caspase-3 cleavages. Furthermore, the U87 glioblastoma tumour-bearing mice treated with LLL-3 exhibited prolonged survival relative to vehicle-treated mice (28.5 vs 16 days) and had smaller intracranial tumours and no evidence of contralateral invasion. These results suggest that LLL-3 may be a potential therapeutic agent in the treatment of glioblastoma with constitutive STAT3 activation. PMID- 19127271 TI - Incidence and survival of childhood bone cancer in northern England and the West Midlands, 1981-2002. AB - There is a paucity of population-based studies examining incidence and survival trends in childhood bone tumours. We used high quality data from four population based registries in England. Incidence patterns and trends were described using Poisson regression. Survival trends were analysed using Cox regression. There were 374 cases of childhood (ages 0-14 years) bone tumours (206 osteosarcomas, 144 Ewing sarcomas, 16 chondrosarcomas, 8 other bone tumours) registered in the period 1981-2002. Overall incidence (per million person years) rates were 2.63 (95% confidence interval (CI) 2.27-2.99) for osteosarcoma, 1.90 (1.58-2.21) for Ewing sarcoma and 0.21 (0.11-0.31) for chondrosarcoma. Incidence of Ewing sarcoma declined at an average rate of 3.1% (95% CI 0.6-5.6) per annum (P=0.04), which may be due to tumour reclassification, but there was no change in osteosarcoma incidence. Survival showed marked improvement over the 20 years (1981-2000) for Ewing sarcoma (hazard ratio (HR) per annum=0.95 95% CI 0.91-0.99; P=0.02). However, no improvement was seen for osteosarcoma patients (HR per annum=1.02 95% CI 0.98-1.05; P=0.35) over this time period. Reasons for failure to improve survival including potential delays in diagnosis, accrual to trials, adherence to therapy and lack of improvement in treatment strategies all need to be considered. PMID- 19127272 TI - Mortality and cancer incidence following occupational radiation exposure: third analysis of the National Registry for Radiation Workers. AB - Mortality and cancer incidence were studied in the National Registry for Radiation Workers in, relative to earlier analyses, an enlarged cohort of 174 541 persons, with longer follow-up (to 2001) and, for the first time, cancer registration data. SMRs for all causes and all malignant neoplasms were 81 and 84 respectively, demonstrating a 'healthy worker effect'. Within the cohort, mortality and incidence from both leukaemia excluding CLL and the grouping of all malignant neoplasms excluding leukaemia increased to a statistically significant extent with increasing radiation dose. Estimates of the trend in risk with dose were similar to those for the Japanese A-bomb survivors, with 90% confidence intervals that excluded both risks more than 2-3 times greater than the A-bomb values and no raised risk. Some evidence of an increasing trend with dose in mortality from all circulatory diseases may, at least partly, be due to confounding by smoking. This analysis provides the most precise estimates to date of mortality and cancer risks following occupational radiation exposure and strengthens the evidence for raised risks from these exposures. The cancer risk estimates are consistent with values used to set radiation protection standards. PMID- 19127270 TI - Vegetable, fruit and antioxidant nutrient consumption and subsequent risk of hepatocellular carcinoma: a prospective cohort study in Japan. AB - In a population-based prospective study of 19,998 Japanese individuals, consumption of vegetables, green-yellow and green leafy vegetables was inversely associated with the risk of hepatocellular carcinoma (101 cases), with multivariable hazard ratios for the highest vs lowest tertile of 0.61 (95% confidence interval (CI)=0.36-1.03, P(trend)=0.07), 0.65 (95% CI=0.39-1.08, P(trend)=0.06) and 0.59 (95% CI=0.35-1.01, P(trend)=0.04), respectively. PMID- 19127273 TI - Cancer in the offspring of female radiation workers: a record linkage study. AB - This study uses record linkage between the National Registry of Childhood Tumours (NRCT) and the National Registry for Radiation Workers to re-assess our earlier finding that the offspring of women radiation workers exposed to ionising radiation before the child's conception may be at an increased risk of childhood cancer. An additional 16,964 childhood cancer patients taken from the NRCT, together with the same number of matched controls, are included. Pooled analyses, based on the new and original datasets, include 52,612 cases and their matched controls. Relative risks (RRs) for maternal employment as a radiation worker, maternal exposure or not during the relevant pregnancy and pattern of employment relative to conception and diagnosis dates were calculated.The new data provide no evidence of an increased risk of childhood cancer associated with maternal preconception radiation work and thus do not support our earlier finding of a raised risk in the offspring of female radiation workers. Considering the pooled data, a weak association was found between maternal radiation work during pregnancy and childhood cancer in offspring although the evidence is limited by the small numbers of linked cases and controls. PMID- 19127274 TI - Estimation of lead time and overdiagnosis in breast cancer screening. PMID- 19127276 TI - Sarcopenia and the elusive fountain of youth. PMID- 19127280 TI - Angiotensin II modulates BBB permeability via activation of the AT(1) receptor in brain endothelial cells. AB - Hypertensive encephalopathy occurs when acute changes in blood pressure cause breakdown of the blood-brain barrier (BBB). Angiotensin II (Ang II) plays a role in this pathophysiology. We determined whether Ang II directly regulates endothelial cell function at the BBB. In BBB microvessel endothelial cells (MECs), the Ang II (100 nmol/L; 0 to 6 h) effects on permeability to (125)I albumin and transendothelial electrical resistance (TEER) were assessed. Angiotensin II (100 nmol/L) caused significant time-dependent changes in both (125)I-albumin permeability (25%) at 2 h and TEER (-8.87 Omega x cm(2)) at 6 h. Next, MECs were pretreated with the Ang II type 1 (AT(1)) receptor blocker telmisartan (1 micromol/L) or the Ang II type 2 (AT(2)) receptor blocker PD123,319 (1 micromol/L) followed by treatment with Ang II (100 nm). Telmisartan completely inhibited the Ang II-induced increase in (125)I-albumin permeability in MECs whereas PD123,319 had no effect. Using western blot analysis, we showed that MECs express AT(1) receptors but not AT(2) receptors. Treatment with Ang II (100 nmol/L; 0 to 6 h) also increased total protein kinase C activity. In contrast, Ang II had no effect on the expression of occludin, claudin 5, or actin. These results show that Ang II directly modulates transcytotic and paracellular permeability in BBB endothelial cells and could contribute to the pathophysiology of hypertensive encephalopathy. PMID- 19127281 TI - Measuring abdominal adiposity in 6 to 7-year-old children. AB - BACKGROUND/OBJECTIVES: Both intra-abdominal adipose tissue (IAAT) and subcutaneous abdominal adipose tissue (SAAT) are associated with cardiovascular risk factors, even in childhood. Currently, the gold standard in assessing IAAT and SAAT is computed tomography (CT), which is not widely applicable. The aim of this study was to estimate abdominal fat using anthropometry, dual-energy X-ray absorptiometry (DEXA) and ultrasound, and compare these estimates with the amounts of IAAT and SAAT determined by CT in 6 to 7-year-old children. SUBJECTS/METHODS: In 31 healthy children, weight, height, circumferences, skinfolds, DEXA, abdominal ultrasound and CT were performed. Measurements were compared by simple correlations and receiver operating characteristic analyses. RESULTS: Total abdominal fat on CT did not differ between boys and girls (86.5 versus 89.8 cm(3), P=0.84). Boys had a higher IAAT to SAAT ratio than girls (0.56 versus 0.37, P=0.03). The sum of supra-iliac and abdominal skinfolds was most strongly correlated with SAAT on CT (r=0.93, P<0.001), and the abdominal skinfold with IAAT on CT (r=0.72, P<0.001). Diagnosis of subcutaneous abdominal and intra abdominal adiposity can also be made using skinfolds. The associations with circumferences, body mass index and DEXA were less pronounced; however, these techniques can also be used to classify children according to SAAT and IAAT. Ultrasound can be used to diagnose subcutaneous adiposity, although it was not superior to skinfold measurements. CONCLUSION: Skinfold measurements are the best non-invasive technique in predicting subcutaneous as well as intra-abdominal fat in our population of 6 to 7-year-old children. PMID- 19127282 TI - Global similarity with local differences in linkage disequilibrium between the Dutch and HapMap-CEU populations. AB - The HapMap project has facilitated the selection of tagging single nucleotide polymorphisms (tagSNPs) for genome-wide association studies (GWAS) under the assumption that linkage disequilibrium (LD) in the HapMap populations is similar to the populations under investigation. Earlier reports support this assumption, although in most of these studies only a few loci were evaluated. We compared pair-wise LD and LD block structure across autosomes between the Dutch population and the CEU-HapMap reference panel. The impact of sampling distribution on the estimation of LD blocks was studied by bootstrapping. A high Pearson correlation (genome-wide; 0.93) between pair-wise r(2) for the Dutch and the CEU populations was found, indicating that tagSNPs from the CEU-HapMap panel capture common variation in the Dutch population. However, some genomic regions exhibited, significantly lower correlation than the genome-wide estimate. This might decrease the validity of HapMap tagSNPs in these regions and the power of GWAS. The LD block structure differed considerably between the Dutch and CEU-HapMap populations. This was not explained by demographic differences between the CEU and Dutch samples, as testing for population stratification was not significant. We also found that sampling variation had a large effect on the estimation of LD blocks, as shown by the bootstrapping analysis. Thus, in small samples, most of the observed differences in LD blocks between populations are most likely the result of sampling variation. This poor concordance in LD block structure suggests that large samples are required for robust estimations of local LD block structure in populations. PMID- 19127283 TI - A simple pharmacokinetic model to characterize exposure of Americans to di-2 ethylhexyl phthalate. AB - A simple pharmacokinetic model to predict concentrations of metabolites of di-2 ethylhexyl phthalate, DEHP, in humans starting from intakes of DEHP was developed and applied. This model predicts serum and urine concentrations of five DEHP metabolites: MEHP, 5oxo-MEHP, 5OH-MEHP, 5cx-MEPP, and 2cx-MMHP. The model was calibrated using data from an individual who dosed himself with 48.5 mg DEHP, and then took blood and urine samples over a 44-h period. The calibrated model was then used in two applications: one on a second set of individuals whose exposure to DEHP was through PVC medical devices in a blood platelet donation procedure, and one on background exposures in the United States (US). Based on 2001/02 NHANES data, median US background urine concentrations of MEHP, 5OH-MEHP, and 5oxo-MEHP are 4.1, 20.1, and 14.0 microg/l, respectively. Creatine and urine volume-correction approaches were used to backcalculate an average daily dose of DEHP in the range of 0.6-2.2 microg/kg per day. A "background cohort" including 8 individuals and 57 complete days of urination were assumed to be exposed to1.5 microg/kg per day, spread out in equal doses of 0.3 microg/kg per day at 0900, 1200, 1500, 1800, and 2100 h. The average predicted urine concentrations were 4.6, 15.9, and 9.4 microg/l for MEHP, 5OH-MEHP, and 5oxo-MEHP. These are similar, but the two secondary metabolites are slightly lower than medians found in NHANES. This slight difference between the NHANES results and the background simulations could have been due to differences in metabolism between the individual who provided the calibration data (61-year-old Caucasian male) and the general US population. Another explanation evaluated was that urine concentrations further from the time of exposure may have larger disparities between MEHP and the two secondary metabolites as compared with concentrations measured closer to the time of exposure. PMID- 19127284 TI - Is 21st century neuroscience too focussed on the rat/mouse model of brain function and dysfunction? AB - Studies in the basic neurosciences are heavily reliant upon rat and mouse models. The brain is one of the most distinguishing features of the human species, but is enough being done to fully understand the evolution of the human brain and brain diversity in general? Without a clear understanding of the evolution of the nervous system we may be investing a great deal of effort into some limited specific animal models that may prove to be erroneous in terms of the overall usefulness in clinically applied research. Here we present an analysis that demonstrates that 75% of our research efforts are directed to the rat, mouse and human brain, or 0.0001% of the nervous systems on the planet. This extreme bias in research trends may provide a limited scope in the discovery of novel aspects of brain structure and function that would be of importance in understanding both the evolution of the human brain and in selecting appropriate animal models for use in clinically related research. We offer examples both from the historical and recent literature indicating the usefulness of comparative neurobiological investigation in elucidating both normal and abnormal structure and function of the brain. PMID- 19127285 TI - Neighbourhood socioeconomics status predicts non-cardiovascular mortality in cardiac patients with access to universal health care. AB - BACKGROUND: Although the Canadian health care system provides essential services to all residents, evidence suggests that socioeconomic gradients in disease outcomes still persist. The main objective of our study was to investigate whether mortality, from cardiovascular disease or other causes, varies by neighbourhood socioeconomic gradients in patients accessing the healthcare system for cardiovascular disease management. METHODS AND FINDINGS: A cohort of 485 patients with angiographic evidence of coronary artery disease (CAD) and neighbourhood socioeconomic status information was followed for 13.3 years. Survival analyses were completed with adjustment for potentially confounding risk factors. There were 64 cases of cardiovascular mortality and 66 deaths from non cardiovascular chronic diseases. No socioeconomic differentials in cardiovascular mortality were observed. However, lower neighbourhood employment, education, and median family income did predict an increased risk of mortality from non cardiovascular chronic diseases. For each quintile decrease in neighbourhood socioeconomic status, non-cardiovascular mortality risk rose by 21-30%. Covariate adjusted hazard ratios (95% confidence interval) for non-cardiovascular mortality were 1.21 (1.02-1.42), 1.21 (1.01-1.46), and 1.30 (1.06-1.60), for each quintile decrease in neighbourhood education, employment, and income, respectively. These patterns were primarily attributable to mortality from cancer. Estimated risks for mortality from cancer rose by 42% and 62% for each one quintile decrease in neighbourhood median income and employment rate, respectively. Although only baseline clinical information was collected and patient-level socioeconomic data were not available, our results suggest that environmental socioeconomic factors have a significant impact on CAD patient survival. CONCLUSIONS: Despite public health care access, CAD patients who reside in lower-socioeconomic neighbourhoods show increased vulnerability to non-cardiovascular chronic disease mortality, particularly in the domain of cancer. These findings prompt further research exploring mechanisms of neighbourhood effects on health, and ways they may be ameliorated. PMID- 19127286 TI - Atmospheric hypoxia limits selection for large body size in insects. AB - BACKGROUND: The correlations between Phanerozoic atmospheric oxygen fluctuations and insect body size suggest that higher oxygen levels facilitate the evolution of larger size in insects. METHODS AND PRINCIPAL FINDINGS: Testing this hypothesis we selected Drosophila melanogaster for large size in three oxygen atmospheric partial pressures (aPO(2)). Fly body sizes increased by 15% during 11 generations of size selection in 21 and 40 kPa aPO(2). However, in 10 kPa aPO(2), sizes were strongly reduced. Beginning at the 12(th) generation, flies were returned to normoxia. All flies had similar, enlarged sizes relative to the starting populations, demonstrating that selection for large size had functionally equivalent genetic effects on size that were independent of aPO(2). SIGNIFICANCE: Hypoxia provided a physical constraint on body size even in a tiny insect strongly selected for larger mass, supporting the hypothesis that Triassic hypoxia may have contributed to a reduction in insect size. PMID- 19127287 TI - The sleeping brain's influence on verbal memory: boosting resistance to interference. AB - Memories evolve. After learning something new, the brain initiates a complex set of post-learning processing that facilitates recall (i.e., consolidation). Evidence points to sleep as one of the determinants of that change. But whenever a behavioral study of episodic memory shows a benefit of sleep, critics assert that sleep only leads to a temporary shelter from the damaging effects of interference that would otherwise accrue during wakefulness. To evaluate the potentially active role of sleep for verbal memory, we compared memory recall after sleep, with and without interference before testing. We demonstrated that recall performance for verbal memory was greater after sleep than after wakefulness. And when using interference testing, that difference was even more pronounced. By introducing interference after sleep, this study confirms an experimental paradigm that demonstrates the active role of sleep in consolidating memory, and unmasks the large magnitude of that benefit. PMID- 19127288 TI - Two host factors regulate persistence of H7-specific T cells injected in tumor bearing mice. AB - BACKGROUND: Injection of CD8 T cells primed against immunodominant minor histocompatibility antigens (MiHA) such as H7(a) can eradicate leukemia and solid tumors. To understand why MiHA-targeted T cells have such a potent antitumor effect it is essential to evaluate their in vivo behavior. In the present work, we therefore addressed two specific questions: what is the proliferative dynamics of H7(a)-specifc T cells in tumors, and do H7(a)-specific T cells persist long term after adoptive transfer? METHODOLOGY/PRINCIPAL FINDINGS: By day 3 after adoptive transfer, we observed a selective infiltration of melanomas by anti H7(a) T cells. Over the next five days, anti-H7(a) T cells expanded massively in the tumor but not in the spleen. Thus, by day 8 after injection, anti-H7(a) T cells in the tumor had undergone more cell divisions than those in the spleen. These data strongly suggest that anti-H7(a) T cells proliferate preferentially and extensively in the tumors. We also found that two host factors regulated long term persistence of anti-H7(a) memory T cells: thymic function and expression of H7(a) by host cells. On day 100, anti-H7(a) memory T cells were abundant in euthymic H7(a)-negative (B10.H7(b)) mice, present in low numbers in thymectomized H7(a)-positive (B10) hosts, and undetectable in euthymic H7(a)-positive recipients. CONCLUSIONS/SIGNIFICANCE: Although in general the tumor environment is not propitious to T-cell invasion and expansion, the present work shows that this limitation may be overcome by adoptive transfer of primed CD8 T cells targeted to an immunodominant MiHA (here H7(a)). At least in some cases, prolonged persistence of adoptively transferred T cells may be valuable for prevention of late cancer relapse in adoptive hosts. Our findings therefore suggest that it may be advantageous to target MiHAs with a restricted tissue distribution in order to promote persistence of memory T cells and thereby minimize the risk of cancer recurrence. PMID- 19127289 TI - Can playing the computer game "Tetris" reduce the build-up of flashbacks for trauma? A proposal from cognitive science. AB - BACKGROUND: Flashbacks are the hallmark symptom of Posttraumatic Stress Disorder (PTSD). Although we have successful treatments for full-blown PTSD, early interventions are lacking. We propose the utility of developing a 'cognitive vaccine' to prevent PTSD flashback development following exposure to trauma. Our theory is based on two key findings: 1) Cognitive science suggests that the brain has selective resources with limited capacity; 2) The neurobiology of memory suggests a 6-hr window to disrupt memory consolidation. The rationale for a 'cognitive vaccine' approach is as follows: Trauma flashbacks are sensory perceptual, visuospatial mental images. Visuospatial cognitive tasks selectively compete for resources required to generate mental images. Thus, a visuospatial computer game (e.g. "Tetris") will interfere with flashbacks. Visuospatial tasks post-trauma, performed within the time window for memory consolidation, will reduce subsequent flashbacks. We predicted that playing "Tetris" half an hour after viewing trauma would reduce flashback frequency over 1-week. METHODOLOGY/PRINCIPAL FINDINGS: The Trauma Film paradigm was used as a well established experimental analog for Post-traumatic Stress. All participants viewed a traumatic film consisting of scenes of real injury and death followed by a 30-min structured break. Participants were then randomly allocated to either a no-task or visuospatial ("Tetris") condition which they undertook for 10-min. Flashbacks were monitored for 1-week. Results indicated that compared to the no task condition, the "Tetris" condition produced a significant reduction in flashback frequency over 1-week. Convergent results were found on a clinical measure of PTSD symptomatology at 1-week. Recognition memory between groups did not differ significantly. CONCLUSIONS/SIGNIFICANCE: Playing "Tetris" after viewing traumatic material reduces unwanted, involuntary memory flashbacks to that traumatic film, leaving deliberate memory recall of the event intact. Pathological aspects of human memory in the aftermath of trauma may be malleable using non-invasive, cognitive interventions. This has implications for a novel avenue of preventative treatment development, much-needed as a crisis intervention for the aftermath of traumatic events. PMID- 19127290 TI - Relatively low HIV infection rates in rural Uganda, but with high potential for a rise: a cohort study in Kayunga District, Uganda. AB - BACKGROUND: Few studies have been conducted in Uganda to identify and quantify the determinants of HIV-1 infection. We report results from a community-based cohort study, whose primary objectives were to determine HIV-1 prevalence, incidence, and determinants of these infections, among other objectives. METHODOLOGY: Consenting volunteers from the rural district of Kayunga in Uganda aged 15-49 years were enrolled between March and July 2006. Participants were evaluated every six months. A questionnaire that collected information on behavioral and other HIV-1 risk factors was administered, and a blood sample obtained for laboratory analysis at each study visit. PRINCIPAL FINDINGS: HIV-1 prevalence among the 2025 participants was 9.9% (95% CI = 8.6%-11.2%). By the end of 12 months of follow-up, 1689.7 person-years had been accumulated, with a median follow-up time of 11.97 months. Thirteen HIV-1 incident cases were detected giving an annual HIV-1 incidence of 0.77% (95% CI = 0.35-1.19). Prevalence of HSV-2 infection was 57% and was strongly associated with prevalent HIV-1 infection (adjusted Odds Ratio = 3.9, 95% CI = 2.50-6.17); as well as incident HIV-1 infection (adjusted Rate Ratio (RR) = 8.7, 95% CI = 1.11-67.2). The single most important behavioral characteristic associated with incident HIV infection was the number of times in the past 6 months, a participant had sex with person(s) they suspected/knew were having sex with others; attaining statistical significance at 10 times and higher (adjusted RR = 6.3, 95% CI = 1.73 23.1). By the end of 12 months of follow-up, 259 participants (13%) were lost to follow-up, 13 (0.6%) had died, and 2 (0.1%) had withdrawn consent. CONCLUSIONS: Despite relatively low HIV-1 incidence observed in this community, prevalence remains relatively high. In the presence of high prevalence of HSV-2 infection and the behavioral characteristic of having sex with more than one partner, there is potential for increase in HIV-1 incidence. PMID- 19127291 TI - Peptides derived from HIV-1 integrase that bind Rev stimulate viral genome integration. AB - BACKGROUND: The human immunodeficiency virus type 1 (HIV-1) integrase protein (IN), catalyzes the integration of viral DNA into the host cell genome. IN catalyzes the first step of the integration process, namely the 3'-end processing in which IN removes a pGT dinucleotide from the 3' end of each viral long terminal repeat (LTR). Following nuclear import of the viral preintegration complex, the host chromosomal DNA becomes accessible to the viral cDNA and the second step of the integration process, namely the strand-transfer step takes place. This ordered sequence of events, centered on integration, is mandatory for HIV replication. METHODOLOGY/PRINCIPAL FINDINGS: Using an integrase peptide library, we selected two peptides, designated INr-1 and INr-2, which interact with the Rev protein and probably mediate the Rev-integrase interaction. Using an in-vitro assay system, we show that INr-1 and INr-2 are able to abrogate the inhibitory effects exerted by Rev and Rev-derived peptides on integrase activity. Both INr-1 and INr-2 were found to be cell-permeable and nontoxic, allowing a study of their effect in HIV-1-infected cultured cells. Interestingly, both INr peptides stimulated virus infectivity as estimated by production of the viral P24 protein, as well as by determination of the appearance of newly formed virus particles. Furthermore, kinetics studies revealed that the cell-permeable INr peptides enhance the integration process, as was indeed confirmed by direct determination of viral DNA integration by real-time PCR. CONCLUSIONS/SIGNIFICANCE: The results of the present study raise the possibility that in HIV-infected cells, the Rev protein may be involved in the integration of proviral DNA by controlling/regulating the activity of the integrase. Release from such inhibition leads to stimulation of IN activity and multiple viral DNA integration events. PMID- 19127292 TI - Diabetes and the risk of multi-system aging phenotypes: a systematic review and meta-analysis. AB - BACKGROUND: Observational studies suggested an association between diabetes and the risk of various geriatric conditions (i.e., cognitive impairment, dementia, depression, mobility impairment, disability, falls, and urinary incontinence). However, the magnitude and impact of diabetes on older adults have not been reviewed. METHODOLOGY/PRINCIPAL FINDINGS: MEDLINE and PSYCINFO databases were searched through November 2007 for published studies, supplemented by manual searches of bibliographies of key articles. Population-based, prospective cohort studies that reported risk of geriatric outcomes in relation to diabetes status at baseline were selected. Two authors independently extracted the data, including study population and follow-up duration, ascertainment of diabetes status at baseline, outcomes of interest and their ascertainment, adjusted covariates, measures of association, and brief results. Fifteen studies examined the association of DM with cognitive dysfunction. DM was associated with a faster decline in cognitive function among older adults. The pooled adjusted risk ratio (RR) for all dementia when persons with DM were compared to those without was 1.47 (95% CI, 1.25 to 1.73). Summary RRs for Alzheimer's disease and vascular dementia comparing persons with DM to those without were 1.39 (CI, 1.16 to 1.66) and 2.38 (CI, 1.79 to 3.18), respectively. Four of 5 studies found significant association of DM with faster mobility decline and incident disability. Two studies examined the association of diabetes with falls in older women. Both found statistically significant associations. Insulin users had higher RR for recurrent falls. One study for urinary incontinence in older women found statistically significant associations. Two studies for depression did not suggest that DM was an independent predictor of incident depression. CONCLUSIONS/SIGNIFICANCE: Current evidence supports that DM is associated with increased risk for selected geriatric conditions. Clinicians should increase their awareness and provide appropriate care. Future research is required to elucidate the underlying pathological pathway. PMID- 19127294 TI - Olfactory sex recognition investigated in Antarctic prions. AB - Chemical signals can yield information about an animal such as its identity, social status or sex. Such signals have rarely been considered in birds, but recent results have shown that chemical signals are actually used by different bird species to find food and to recognize their home and nest. This is particularly true in petrels whose olfactory anatomy is among the most developed in birds. Recently, we have demonstrated that Antarctic prions, Pachyptila desolata, are also able to recognize and follow the odour of their partner in a Y maze.However, the experimental protocol left unclear whether this choice reflected an olfactory recognition of a particular individual (i.e. partner) or a more general sex recognition mechanism. To test this second hypothesis, male and female birds' odours were presented simultaneously to 54 Antarctic prions in a Y maze. Results showed random behaviour by the tested bird, independent of its sex or reproductive status. Present results do not support the possibility that Antarctic prions can distinguish the sex of a conspecific through its odour but indirectly support the hypothesis that they can distinguish individual odours. PMID- 19127293 TI - Gonadal transcriptome alterations in response to dietary energy intake: sensing the reproductive environment. AB - Reproductive capacity and nutritional input are tightly linked and animals' specific responses to alterations in their physical environment and food availability are crucial to ensuring sustainability of that species. We have assessed how alterations in dietary energy intake (both reductions and excess), as well as in food availability, via intermittent fasting (IF), affect the gonadal transcriptome of both male and female rats. Starting at four months of age, male and female rats were subjected to a 20% or 40% caloric restriction (CR) dietary regime, every other day feeding (IF) or a high fat-high glucose (HFG) diet for six months. The transcriptional activity of the gonadal response to these variations in dietary energy intake was assessed at the individual gene level as well as at the parametric functional level. At the individual gene level, the females showed a higher degree of coherency in gonadal gene alterations to CR than the males. The gonadal transcriptional and hormonal response to IF was also significantly different between the male and female rats. The number of genes significantly regulated by IF in male animals was almost 5 times greater than in the females. These IF males also showed the highest testosterone to estrogen ratio in their plasma. Our data show that at the level of gonadal gene responses, the male rats on the IF regime adapt to their environment in a manner that is expected to increase the probability of eventual fertilization of females that the males predict are likely to be sub-fertile due to their perception of a food deficient environment. PMID- 19127295 TI - Preferences across the menstrual cycle for masculinity and symmetry in photographs of male faces and bodies. AB - BACKGROUND: Previous studies have shown that women increase their preference for masculinity during the fertile phase of the menstrual cycle. Evidence for a similar preference shift for symmetry is equivocal. These studies have required participants to choose between subtle variations in computer-generated stimuli, and preferences for more natural stimuli have not been investigated. METHODOLOGY/PRINCIPAL FINDINGS: Our study employed photographs of individual males to investigate women's preferences for face and body masculinity and symmetry across the menstrual cycle. We collected attractiveness ratings from 25 normally cycling women at high- and low-fertility days of the menstrual cycle. Attractiveness ratings made by these women were correlated with independent ratings of masculinity and symmetry provided by different sets of raters. We found no evidence for any cyclic shift in female preferences. Correlations between attractiveness and masculinity, and attractiveness and symmetry did not differ significantly between high- and low-fertility test sessions. Furthermore, there was no significant difference between high- and low-fertility ratings of attractiveness. CONCLUSIONS: These results suggest that a menstrual cycle shift in visual preferences for masculinity and symmetry may be too subtle to influence responses to real faces and bodies, and subsequent mate-choice decisions. PMID- 19127296 TI - In-vitro helix opening of M. tuberculosis oriC by DnaA occurs at precise location and is inhibited by IciA like protein. AB - BACKGROUND: Mycobacterium tuberculosis (M.tb), the pathogen that causes tuberculosis, is capable of staying asymptomatically in a latent form, persisting for years in very low replicating state, before getting reactivated to cause active infection. It is therefore important to study M.tb chromosome replication, specifically its initiation and regulation. While the region between dnaA and dnaN gene is capable of autonomous replication, little is known about the interaction between DnaA initiator protein, oriC origin of replication sequences and their negative effectors of replication. METHODOLOGY/PRINCIPAL FINDINGS: By KMnO(4) mapping assays the sequences involved in open complex formation within oriC, mediated by M.tb DnaA protein, were mapped to position -500 to -518 with respect to the dnaN gene. Contrary to E. coli, the M.tb DnaA in the presence of non-hydrolysable analogue of ATP (ATPgammaS) was unable to participate in helix opening thereby pointing to the importance of ATP hydrolysis. Interestingly, ATPase activity in the presence of supercoiled template was higher than that observed for DnaA box alone. M.tb rRv1985c, a homologue of E.coli IciA (Inhibitor of chromosomal initiation) protein, could inhibit DnaA-mediated in-vitro helix opening by specifically binding to A+T rich region of oriC, provided the open complex formation had not initiated. rIciA could also inhibit in-vitro replication of plasmid carrying the M.tb origin of replication. CONCLUSIONS/SIGNIFICANCE: These results have a bearing on the functional role of the important regulator of M.tb chromosomal replication belonging to the LysR family of bacterial regulatory proteins in the context of latency. PMID- 19127297 TI - Families and Assisted Living. AB - PURPOSE: Despite growing research on assisted living (AL) as a residential care option for older adults, the social ramifications of residents' transitions to assisted living is relatively unexplored. This article examines family involvement in AL, including family structures of residents, types of involvement from family members living outside the AL, and outcomes for these family members. DESIGN AND METHODS: We reviewed current literature utilizing the MEDLINE, PSYCINFO, and CINAHL databases to identify AL studies that examined issues pertaining to families or informal care. Following the screening of abstracts, 180 reports were retrieved for further review, and 62 studies were selected for inclusion. RESULTS: Families visit residents frequently and provide a wide range of instrumental assistance but provide only minimal personal care. Studies of family outcomes indicated relatively high satisfaction, but potential care burden as well. IMPLICATIONS: How family care and involvement occurs in AL in relation to formal care provision and whether various types of formal-informal care integration influence family outcomes remains unclear. We suggest a research agenda that attempts to tease out causal relationships for family involvement, differentiate family roles, and implement longitudinal analyses for a range of family outcomes. PMID- 19127298 TI - Molecular identification of birds: performance of distance-based DNA barcoding in three genes to delimit parapatric species. AB - BACKGROUND: DNA barcoding based on the mitochondrial cytochrome oxidase subunit I gene (cox1 or COI) has been successful in species identification across a wide array of taxa but in some cases failed to delimit the species boundaries of closely allied allopatric species or of hybridising sister species. METHODOLOGY/PRINCIPAL FINDINGS: In this study we extend the sample size of prior studies in birds for cox1 (2776 sequences, 756 species) and target especially species that are known to occur parapatrically, and/or are known to hybridise, on a Holarctic scale. In order to obtain a larger set of taxa (altogether 2719 species), we include also DNA sequences of two other mitochondrial genes: cytochrome b (cob) (4614 sequences, 2087 species) and 16S (708 sequences, 498 species). Our results confirm the existence of a wide gap between intra- and interspecies divergences for both cox1 and cob, and indicate that distance-based DNA barcoding provides sufficient information to identify and delineate bird species in 98% of all possible pairwise comparisons. This DNA barcoding gap was not statistically influenced by the number of individuals sequenced per species. However, most of the hybridising parapatric species pairs have average divergences intermediate between intraspecific and interspecific distances for both cox1 and cob. CONCLUSIONS/SIGNIFICANCE: DNA barcoding, if used as a tool for species discovery, would thus fail to identify hybridising parapatric species pairs. However, most of them can probably still assigned to known species by character-based approaches, although development of complementary nuclear markers will be necessary to account for mitochondrial introgression in hybridising species. PMID- 19127299 TI - Explicit logic circuits discriminate neural states. AB - The magnitude and apparent complexity of the brain's connectivity have left explicit networks largely unexplored. As a result, the relationship between the organization of synaptic connections and how the brain processes information is poorly understood. A recently proposed retinal network that produces neural correlates of color vision is refined and extended here to a family of general logic circuits. For any combination of high and low activity in any set of neurons, one of the logic circuits can receive input from the neurons and activate a single output neuron whenever the input neurons have the given activity state. The strength of the output neuron's response is a measure of the difference between the smallest of the high inputs and the largest of the low inputs. The networks generate correlates of known psychophysical phenomena. These results follow directly from the most cost-effective architectures for specific logic circuits and the minimal cellular capabilities of excitation and inhibition. The networks function dynamically, making their operation consistent with the speed of most brain functions. The networks show that well-known psychophysical phenomena do not require extraordinarily complex brain structures, and that a single network architecture can produce apparently disparate phenomena in different sensory systems. PMID- 19127300 TI - Allele-specific gene expression is widespread across the genome and biological processes. AB - Allelic specific gene expression (ASGE) appears to be an important factor in human phenotypic variability and as a consequence, for the development of complex traits and diseases. In order to study ASGE across the human genome, we have performed a study in which genotyping was coupled with an analysis of ASGE by screening 11,500 SNPs using the Mapping 10 K Array to identify differential allelic expression. We found that from the 5,133 SNPs that were suitable for analysis (heterozygous in our sample and expressed in peripheral blood mononuclear cells), 2,934 (57%) SNPs had differential allelic expression. Such SNPs were equally distributed along human chromosomes and biological processes. We validated the presence or absence of ASGE in 18 out 20 SNPs (90%) randomly selected by real time PCR in 48 human subjects. In addition, we observed that SNPs close to -but not included in- segmental duplications had increased levels of ASGE. Finally, we found that transcripts of unknown function or non-coding RNAs, also display ASGE: from a total of 2,308 intronic SNPs, 1510 (65%) SNPs underwent differential allelic expression. In summary, ASGE is a widespread mechanism in the human genome whose regulation seems to be far more complex than expected. PMID- 19127301 TI - PEG minocycline-liposomes ameliorate CNS autoimmune disease. AB - BACKGROUND: Minocycline is an oral tetracycline derivative with good bioavailability in the central nervous system (CNS). Minocycline, a potent inhibitor of matrix metalloproteinase (MMP)-9, attenuates disease activity in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Potential adverse effects associated with long-term daily minocycline therapy in human patients are concerning. Here, we investigated whether less frequent treatment with long-circulating polyethylene glycol (PEG) minocycline liposomes are effective in treating EAE. FINDINGS: Performing in vitro time kinetic studies of PEG minocycline-liposomes in human peripheral blood mononuclear cells (PBMCs), we determined that PEG minocycline-liposome preparations stabilized with CaCl(2) are effective in diminishing MMP-9 activity. Intravenous injections of PEG minocycline-liposomes every five days were as effective in ameliorating clinical EAE as daily intraperitoneal injections of minocycline. Treatment of animals with PEG minocycline-liposomes significantly reduced the number of CNS-infiltrating leukocytes, and the overall expression of MMP-9 in the CNS. There was also a significant suppression of MMP-9 expression and proteolytic activity in splenocytes of treated animals, but not in CNS infiltrating leukocytes. Thus, leukocytes gaining access to the brain and spinal cord require the same absolute amount of MMP-9 in all treatment groups, but minocycline decreases the absolute cell number. CONCLUSIONS: Our data indicate that less frequent injections of PEG minocycline-liposomes are an effective alternative pharmacotherapy to daily minocycline injections for the treatment of CNS autoimmune diseases. Also, inhibition of MMP-9 remains a promising treatment target in EAE and patients with MS. PMID- 19127302 TI - Endemicity, biogeograhy, composition, and community structure on a northeast pacific seamount. AB - The deep ocean greater than 1 km covers the majority of the earth's surface. Interspersed on the abyssal plains and continental slope are an estimated 14000 seamounts, topographic features extending 1000 m off the seafloor. A variety of hypotheses are posited that suggest the ecological, evolutionary, and oceanographic processes on seamounts differ from those governing the surrounding deep sea. The most prominent and oldest of these hypotheses, the seamount endemicity hypothesis (SMEH), states that seamounts possess a set of isolating mechanisms that produce highly endemic faunas. Here, we constructed a faunal inventory for Davidson Seamount, the first bathymetric feature to be characterized as a 'seamount', residing 120 km off the central California coast in approximately 3600 m of water (Fig 1). We find little support for the SMEH among megafauna of a Northeast Pacific seamount; instead, finding an assemblage of species that also occurs on adjacent continental margins. A large percentage of these species are also cosmopolitan with ranges extending over much of the Pacific Ocean Basin. Despite the similarity in composition between the seamount and non-seamount communities, we provide preliminary evidence that seamount communities may be structured differently and potentially serve as source of larvae for suboptimal, non-seamount habitats. PMID- 19127303 TI - Giant panda genomic data provide insight into the birth-and-death process of mammalian major histocompatibility complex class II genes. AB - To gain an understanding of the genomic structure and evolutionary history of the giant panda major histocompatibility complex (MHC) genes, we determined a 636,503 bp nucleotide sequence spanning the MHC class II region. Analysis revealed that the MHC class II region from this rare species contained 26 loci (17 predicted to be expressed), of which 10 are classical class II genes (1 DRA, 2 DRB, 2 DQA, 3 DQB, 1 DYB, 1 DPA, and 2 DPB) and 4 are non-classical class II genes (1 DOA, 1 DOB, 1 DMA, and 1 DMB). The presence of DYB, a gene specific to ruminants, prompted a comparison of the giant panda class II sequence with those of humans, cats, dogs, cattle, pigs, and mice. The results indicated that birth and death events within the DQ and DRB-DY regions led to major lineage differences, with absence of these regions in the cat and in humans and mice respectively. The phylogenetic trees constructed using all expressed alpha and beta genes from marsupials and placental mammals showed that: (1) because marsupials carry loci corresponding to DR, DP, DO and DM genes, those subregions most likely developed before the divergence of marsupials and placental mammals, approximately 150 million years ago (MYA); (2) conversely, the DQ and DY regions must have evolved later, but before the radiation of placental mammals (100 MYA). As a result, the typical genomic structure of MHC class II genes for the giant panda is similar to that of the other placental mammals and corresponds to BTNL2 approximately DR1 approximately DQ approximately DR2 approximately DY approximately DO_box approximately DP approximately COL11A2. Over the past 100 million years, there has been birth and death of mammalian DR, DQ, DY, and DP genes, an evolutionary process that has brought about the current species-specific genomic structure of the MHC class II region. Furthermore, facing certain similar pathogens, mammals have adopted intra-subregion (DR and DQ) and inter-subregion (between DQ and DP) convergent evolutionary strategies for their alpha and beta genes, respectively. PMID- 19127304 TI - Population dynamics constrain the cooperative evolution of cross-feeding. AB - Cross-feeding is the exchange of nutrients among species of microbes. It has two potential evolutionary origins, one as an exchange of metabolic wastes or byproducts among species, the other as a form of cooperation known as reciprocal altruism. This paper explores the conditions favoring the origin of cooperative cross-feeding between two species. There is an extensive literature on the evolution of cooperation, and some of the requirements for the evolution of cooperative cross-feeding follow from this prior work-specifically the requirement that interactions be limited to small groups of individuals, such as colonies in a spatially structured environment. Evolution of cooperative cross feeding by a species also requires that cross-feeding from the partner species already exists, so that the cooperating mutant will automatically be reciprocated for its actions. Beyond these considerations, some unintuitive dynamical constraints apply. In particular, the benefit of cooperative cross-feeding applies only in the range of intermediate cell densities. At low density, resource concentrations are too low to offset the cost of cooperation. At high density, resources shared by both species become limiting, and the two species become competitors. These considerations suggest that the evolution of cooperative cross-feeding in nature may be more challenging than for other types of cooperation. However, the principles identified here may enable the experimental evolution of cross-feeding, as born out by a recent study. PMID- 19127305 TI - Effects of in utero antiepileptic drug exposure. AB - Recent studies demonstrate an increased teratogenic risk for valproate and a probable increased risk for phenobarbital. Carbamazepine and lamotrigine appear relatively safe; however, results are inconclusive concerning a specific risk for cleft lip/palate for both drugs as well as a dose-dependent effect for malformations associated with lamotrigine. Data regarding teratogenic risks for other antiepileptic drugs are inadequate. Additional studies are needed to delineate further the risks for all antiepileptic drugs and determine the underlying mechanisms. PMID- 19127306 TI - When should clinicians worry about bone density for patients with epilepsy? PMID- 19127307 TI - Is seizure surgery an option for patients with very low IQ? PMID- 19127308 TI - A frontal assault on the generalized nature of juvenile myoclonic epilepsy. PMID- 19127309 TI - Diagnosing pseudoseizures: don't hold your breath. PMID- 19127310 TI - Name calling in the temporal lobe. PMID- 19127311 TI - Do we have a cure for tuberous sclerosis complex? AB - The recent development of several mouse models for tuberous sclerosis complex (TSC) provides in vivo systems to test new therapies for the neurological manifestations of TSC. Rapamycin is known to antagonize the effects of loss of TSC protein function in vitro and in mouse TSC models, rapamycin can prevent seizures and improve learning task performance. These findings provide new hope for TSC patients suffering from intractable seizures and possibly, for those with autism and cognitive disabilities. PMID- 19127312 TI - "TOR"rents of excitement over rapamycin's antiepileptogenic potential. PMID- 19127313 TI - When it comes to GABAergic responses and neonatal seizures--sex matters! PMID- 19127314 TI - The spine loss paradox: clues to mechanisms and meaning. PMID- 19127315 TI - Selective changes in hippocampal GABAA receptors during status epilepticus. PMID- 19127316 TI - Expected and unexpected cardiac problems during pregnancy. PMID- 19127317 TI - Pregnancy in women with prosthetic heart valves. AB - Pregnancy in women with mechanical valve prostheses has a high maternal complication rate including valve thrombosis and death. Coumarin derivatives are relatively safe for the mother with a lower incidence of valve thrombosis than un fractionated and low-molecular-weight heparin, but carry the risk of embryopathy, which is probably dose-dependent. The different anticoagulation regimens are discussed in this review. When valve thrombosis occurs during pregnancy, thrombolysis is the preferable therapeutic option. Bioprostheses have a more favourable pregnancy outcome than mechanical prostheses but due to the high re operation rate in young women they do not constitute the ideal alternative. When women with native valve stenosis need pre-pregnancy intervention, mitral balloon valvuloplasty is the best option in mitral stenosis, while the Ross operation or homograft implantation may be the preferable surgical regimen in aortic stenosis. (Neth Heart J 2008;16:406-11.). PMID- 19127318 TI - Spontaneous coronary artery dissection in the postpartum period. AB - Spontaneous coronary artery dissection is a very uncommon cause of acute coronary syndrome. It occurs predominantly in young to middle-aged women during or after pregnancy. The aetiology remains uncertain. Possible factors are hormonal changes, haemodynamic stress and changes in autoimmune status. In case of single vessel dissection and normal blood flow, conservative treatment often leads to complete angiographic resolution. This case report describes the clinical presentation, diagnosis and therapy of spontaneous coronary artery dissection in a 37-year-old woman in the postpartum period. (Neth Heart J 2008;16:412-4.). PMID- 19127319 TI - A serious complication in the puerperium: peripartum cardiomyopathy. AB - Two women, aged 27, presented with different symptoms shortly after giving birth to their first child. Peripartum cardiomyopathy (PPCM) was diagnosed. PPCM is a rare form of cardiac failure occurring late in pregnancy or in the postpartum period. Many women experience dyspnoea, fatigue, and pedal oedema in the last month of pregnancy or postpartum, symptoms which are identical to early congestive heart failure. Therefore, the diagnosis of PPCM requires vigilance. A high mortality rate and overall poor clinical outcome has been reported in a high percentage of these patients. Subsequent pregnancies remain controversial. (Neth Heart J 2008;16:415-8.). PMID- 19127320 TI - A case of an acquired high-degree AV block in a pregnant woman. AB - Acquired heart block during pregnancy is rare. We describe the case of a 29-year old pregnant female with fatigue due to an unexplained high-degree AV conduction disorder. She was treated with a dual chamber pacing system. Further pregnancy and delivery were without complications. The sparse literature on this topic is reviewed. (Neth Heart J 2008;16:419-21.). PMID- 19127321 TI - Pregnancy and the risk of torsades de pointes in congenital long-QT syndrome. AB - Patients with congenital long-QT syndrome (LQTS) are at increased risk of ventricular arrhythmias during stressful situations. Large-scale studies have pointed out that affected individuals are particularly at risk in the period following pregnancy (post-partum). This is recognised especially for women with an LQTS type 2. Here, we describe two cases of young women with LQTS type 2, both admitted to our institution with symptomatic torsades de pointes a few weeks after delivery. Both patients carried a mutation in the KCNH2 gene. One patient was nullipara, while the other had had an uneventful previous pregnancy. In both cases treatment with a beta-blocker did not prevent life-threatening cardiac arrhythmias. The risk of arrhythmias is thought to gradually decrease to pre pregnancy values in the nine months after delivery. Considering the difficulties related to continuous monitoring of a patient for such a long period and the desire of these patients to have more children in the foreseeable future, ICD implantation was performed. (Neth Heart J 2008;16:422-5.). PMID- 19127322 TI - Left ventricular assist device as a bridge to recovery in a young woman admitted with peripartum cardiomyopathy. AB - Left ventricular assist devices (LVAD) are an effective therapeutic option for end-stage heart failure patients as a bridge to cardiac transplantation in those who deteriorate despite maximal therapy and when a donor heart is not ready available. In some patients, cardiac recovery has been reported while supported by an LVAD. In this case report, we describe a 29-year-old female who was admitted to our centre because of peripartum cardiomyopathy (PPCM). Despite intensive treatment with intravenous inotropes and intra-aortic balloon counter pulsation she had a persisting low cardiac index and an LVAD was implanted. In the months following implantation the left ventricular systolic function improved and the left ventricular dimensions normalised. Eventually the LVAD could be ex planted nine months after implantation. At this moment, three years after explantation, echo-cardiography shows a normal-sized left ventricle and almost completely recovered systolic function. (Neth Heart J 2008;16:426-8). PMID- 19127323 TI - Retrocardiac mass causing collapse. PMID- 19127324 TI - Bone marrow cell therapy after acute myocardial infarction: the HEBE trial in perspective, first results. AB - During the last decennium, the role of bone marrow mononuclear cells (BMMC) has been underscored in the healing process after acute myocardial infarction (AMI). Although these cells improve left ventricular recovery after AMI in experimental studies, results from large-scale randomised trials investigating BMMC therapy in patients with AMI have shown contradictory results. To address this issue the HEBE study was designed, a multicentre, randomised trial, evaluating the effects of intracoronary infusion of BMMCs and the effects of intracoronary infusion of peripheral blood mononuclear cells after primary percutaneous coronary intervention. The primary endpoint of the HEBE trial is the change in regional myocardial function in dysfunctional segments at four months relative to baseline, based on segmental analysis as measured by magnetic resonance imaging. The results from the HEBE trial will provide detailed information about the effects of intracoronary BMMC therapy on post-infarct left ventricular recovery. In addition, further analysis of the data and material obtained may provide important mechanistic insights into the contribution of BMMCs to natural recovery from AMI as well as the response to cell therapy. This may significantly contribute to the development of improved cell-based therapies, aiming at optimising post-infarct recovery and preventing heart failure. (Neth Heart J 2008;16:436-9.). PMID- 19127325 TI - Interaction between parasitophorous vacuolar membrane-associated GRA3 and calcium modulating ligand of host cell endoplasmic reticulum in the parasitism of Toxoplasma gondii. AB - A monoclonal antibody against Toxoplasma gondii of Tg556 clone (Tg556) blotted a 29 kDa protein, which was localized in the dense granules of tachyzoites and secreted into the parasitophorous vacuolar membrane (PVM) after infection to host cells. A cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg556, and the full-length was completed by 5'-RACE of 2,086 bp containing an open reading frame (ORF) of 669 bp. The ORF encoded a polypeptide of 222 amino acids homologous to the revised GRA3 but not to the first reported one. The polypeptide has 3 hydrophobic moieties of an N-terminal stop transfer sequence and 2 transmembrane domains (TMD) in posterior half of the sequence, a cytoplasmic localization motif after the second TMD and an endoplasmic reticulum (ER) retrival motif in the C-terminal end, which suggests GRA3 as a type III transmembrane protein. With the ORF of GRA3, yeast two-hybrid assay was performed in HeLa cDNA expression library, which resulted in the interaction of GRA3 with calcium modulating ligand (CAMLG), a type II transmembrane protein of ER. The specific binding of GRA3 and CAMLG was confirmed by glutathione S-transferase (GST) pull-down and immunoprecipitation assays. The localities of fluorescence transfectionally expressed from GRA3 and CAMLG plasmids were overlapped completely in HeLa cell cytoplasm. In immunofluorescence assay, GRA3 and CAMLG were shown to be co-localized in the PVM of host cells. Structural binding of PVM-inserted GRA3 to CAMLG of ER suggested the receptor ligand of ER recruitment to PVM during the parasitism of T. gondii. PMID- 19127326 TI - Contact-independent cell death of human microglial cells due to pathogenic Naegleria fowleri trophozoites. AB - Free-living Naegleria fowleri leads to a fatal infection known as primary amebic meningoencephalitis in humans. Previously, the target cell death could be induced by phagocytic activity of N. fowleri as a contact-dependent mechanism. However, in this study we investigated the target cell death under a non-contact system using a tissue-culture insert. The human microglial cells, U87MG cells, co cultured with N. fowleri trophozoites for 30 min in a non-contact system showed morphological changes such as the cell membrane destruction and a reduction in the number. By fluorescence-activated cell sorter (FACS) analysis, U87MG cells co cultured with N. fowleri trophozoites in a non-contact system showed a significant increase of apoptotic cells (16%) in comparison with that of the control or N. fowleri lysate. When U87MG cells were co-cultured with N. fowleri trophozoites in a non-contact system for 30 min, 2 hr, and 4 hr, the cytotoxicity of amebae against target cells was 40.5, 44.2, and 45.6%, respectively. By contrast, the cytotoxicity of non-pathogenic N. gruberi trophozoites was 10.2, 12.4, and 13.2%, respectively. These results suggest that the molecules released from N. fowleri in a contact-independent manner as well as phagocytosis in a contact-dependent manner may induce the host cell death. PMID- 19127327 TI - Biochemical, hematological, and electrocardiographic changes in buffaloes naturally infected with Theileria annulata. AB - Changes in selected blood and serum components and electrocardiography (ECG) were investigated in 20 adults (13 females and 7 males) of water buffaloes suffering from severe theileriosis. The age of all animals used in this study ranged 1.5-5 yr. Theileriosis was diagnosed by observation of parasites in the peripheral blood and the presence of schizonts in lymphocytes that were provided from swollen lymph nodes. Statistically significant decreases were observed in the means of RBC, WBC, and packed cell volume (PCV) in blood of infected animals. The means levels of sodium, calcium, phosphorus, and potassium of infected animals were lower than healthy animals, but only the decrease of potassium was significant. The mean serum activities of aspartate transferase and alanine aminotransferase were significantly higher than in uninfected animals. Three cases had atrial premature beat, 2 cases had sinus tachycardia, 2 had sinus arrhythmia, and 1 had first degree of atrioventricular block in ECG. The present study showed that T. annulata infection in cattle is associated with hematological and biochemical, and ECG changes. PMID- 19127328 TI - Sarcocystis and its complications in camels (Camelus dromedarius) of eastern provinces of Iran. AB - The prevalence of Sarcocystis spp. was investigated by gross and histopathological examinations in 250 camels (Camelus dromedarius) slaughtered from 2002 to 2005 in the Mashhad Slaughterhouse, eastern Iran. Samples were taken from the diaphragm, heart, tongue, esophagus and masseter muscles for histopathological studies. No macroscopic sarcocysts were found in the samples at gross inspection. Sarcocysts were detected in 209 of 250 (83.6%) examined camels at histopathological level. The infection rate of the esophagus, heart, masseter muscles, diaphragm, and tongue was 58.8%, 48.0%, 46.8%, 41.6%, and 28.0%, respectively. There was no significant difference in the rate of infection between male (85.8%) and female (81.0%) camels. The tissue response to vital cysts was minimal; however, reaction to the degenerating cysts was severe and caused tissue damages resulting in hyperemia, hemorrhages, mononuclear cell infiltration, necrotic changes, and fibrosis. The wild and domestic carnivores especially dogs may be the final hosts of Sarcocystis spp. in this area. PMID- 19127329 TI - Sympatric distribution of three human Taenia tapeworms collected between 1935 and 2005 in Korea. AB - Taeniasis has been known as one of the prevalent parasitic infections in Korea. Until recently, Taenia saginata had long been considered a dominant, and widely distributed species but epidemiological profiles of human Taenia species in Korea still remain unclear. In order to better understand distribution patterns of human Taenia tapeworms in Korea, partial nucleotide sequences of mitochondrial cox1 and ITS2 (internal transcribed spacer 2) were determined, along with morphological examinations, on 68 Taenia specimens obtained from university museum collections deposited since 1935. Genomic DNA was extracted from formalin preserved specimens. Phylogenetic relationships among the genotypes (cox1 haplotype) detected in this study were inferred using the neighbor-joining method as a tree building method. Morphological and genetic analyses identified 3 specimens as T. solium, 51 specimens as T. asiatica, and 14 specimens as T. saginata. Our results indicate that all 3 Taenia tapeworms are sympatrically distributed in Korea with T. asiatica dominating over T. saginata and T. solium. PMID- 19127330 TI - Reactivity of German cockroach allergen, Bla g 2, peptide fragments to IgE antibodies in patients' sera. AB - Bla g 2 is a cockroach allergen of great importance. This study was conducted to identify IgE-binding epitope(s) of Bla g 2 using the recombinant protein technique. Approximately 50% of tested sera showed IgE reactivity to Pichia expressed Bla g 2 (PrBla g 2) and E. coli-expressed Bla g 2 (ErBla g 2). Only 5.3% of serum samples showed stronger reactivity to PrBla g 2 than ErBla g 2, indicating that serum was reactive to conformational or carbohydrate epitopes. The full-length and 5 peptide fragments of Bla g 2 were produced in E. coli. All fragments showed IgE-binding activity to the cockroach-allergy patients' sera. Specifically, peptide fragments of amino acid residue 1-75 and 146-225 appeared to be important for IgE-binding. The information about the IgE-binding epitope of Bla g 2 can aid in the diagnosis and treatment for cockroach allergies. PMID- 19127331 TI - Infection status of freshwater fish with metacercariae of Clonorchis sinensis in Korea. AB - This study investigated freshwater fish for their current infection status with metacercariae of Clonorchis sinensis in Korea. Twenty-one species of freshwater fish (n = 677) were collected from 34 regions nationwidely from February 2007 to June 2008. They were individually examined by digestion technique. Eight species of freshwater fish from 17 different regions were recognized positive for the metacercariae of C. sinensis. The positive rates (range of metacercariae number per fish) of fish by the species were as follows: 48% (1-1,142) in Pseudorasbora parva, 60% (1-412) in Pungtungia herzi, 15.7% (1-23) in Pseudogobio esocinus, 29% (1-7) in Acheilognathus intermedia, 21% (1-4) in Odontobutis interrupta, 33% (1 6) in Zacco temmincki, 3.6% (1-4) in Zacco platypus, and 26.3% (1) in Hemibarbus labeo. The two species, P. parva and P. herzi, are able to be the index fish for estimation of C. sinensis transmission in a certain locality. Still several species of freshwater fish are briskly transmitting C. sinensis infection in many riverside areas of southern Korea. PMID- 19127332 TI - Fishborne trematode metacercariae detected in freshwater fish from Vientiane Municipality and Savannakhet Province, Lao PDR. AB - Freshwater fish from Vientiane Municipality and Savannakhet Province, Lao PDR were examined by the muscle compression and artificial digestion methods to know the infection status with trematode metacercariae. In the fish from Savannakhet, 2 species of metacercariae, Opisthorchis viverrini and Haplorchis taichui, were detected. O. viverrini metacercariae were found in 6 species of fish, Puntius brevis, Hampala dispar, Esomus metallicus, Mystacoleucus marginatus, Puntioplites falcifer, and Cyclocheilichthys armatus. H. taichui metacercariae were detected in 3 species of fish, P. brevis, P. falcifer, and M. marginatus. In the fish from Vientiane, 4 species of metacercariae, O. viverrini, H. taichui, Haplorchis yokogawai, and Centrocestus formosanus, were detected. Among them, O. viverrini metacercariae were found in 7 species of fish, Onychostoma elongatum, C. armatus, H. dispar, P. brevis, Cyclocheilichthys repasson, Osteochilus hasseltii, and Hypsibarbus lagleri. The metacercariae of H. taichui were detected in 6 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, and Cirrhinus molitorella. The metacercariae of H. yokogawai were found in 9 species of fish, C. repasson, O. elongatum, C. armatus, H. dispar, Labiobarbus leptocheila, O. hasseltii. C. molitorella, Hypsibarbus wetmorei, and H. lagleri. The metacercariae of C. formosanus were detected in 4 species of fish, C. repasson, P. brevis, O. hasseltii, and C. molitorella. From these results, it is confirmed that fishborne trematode metacercariae, i.e. O. viverrini, H. taichui, H. yokogawai and C. formosanus, are prevalent in various species of freshwater fish from Savannakhet Province and Vientiane Municipality, Lao PDR. PMID- 19127333 TI - A case of fatal strongyloidiasis in a patient with chronic lymphocytic leukemia and molecular characterization of the isolate. AB - Strongyloides stercoralis is a human intestinal parasite which may lead to complicated strongyloidiasis in immunocompromised. Here, a case of complicated strongyloidiasis in a patient with chronic lymphocytic leukemia is reported. Presence of numerous S. stercoralis larvae in feces and sputum confirmed the diagnosis of hyperinfection syndrome in this patient. Following recovery of filariform larvae from agar plate culture of the stool, the isolate was characterized for the ITS1 region of ribosomal DNA gene by nested-PCR and sequencing. Albendazole therapy did not have cure effects; and just at the beginning of taking ivermectin, the patient died. The most important clue to prevent such fatal consequences is early diagnosis and proper treatment. PMID- 19127334 TI - Periocular dirofilariasis in a young woman: a case report. AB - A case of periocular dirofilariasis has been diagnosed in Iran. A 27-yr old female referred with a history of edema and redness in the left eye since 2 wk ago. On slit lamp examination, a moving thread-like worm was seen in the subconjunctival area. Two days later, a 126 mm long, live filaria worm was extracted from the lower lid subcutaneous tissue. The worm was diagnosed as, likelihood, Dirofilaria immitis on microscopic examinations. The present case suggests that D. immitis can cause periocular infections. PMID- 19127335 TI - Reemergence of the bedbug Cimex lectularius in Seoul, Korea. AB - A healthy 30-yr-old woman carrying an insect that had been caught in her living room visited the International Clinic at Severance Hospital, Seoul, in December 2007. The insect she brought was identified to be a nymph of a bedbug, Cimex lectularius, and her skin rashes looked typical bedbug's bites. Her apartment was investigated, and a dead body of a bedbug, cast skins, and hatched eggs were found in her rooms and neighbors' rooms in the same building. She was living in that apartment in Seoul for 9 months since she had moved from New Jersey, USA. We assume that the bedbugs were introduced from abroad, since there had been no report on bedbugs in Seoul for more than 2 decades at least. This is a report of a reemergence of the common bedbug, C. lectularius in Seoul, Korea. PMID- 19127336 TI - The first outbreak of Chorioptes texanus (Acari: Psoroptidae) infestation in a cattle farm in Korea. AB - Mites in the genus Chorioptes cause a mild form of skin disease in both domestic and wild ruminants. In July 2006, dermatitis characterized by alopecia, marked lichenification, accumulation of crust, and fissuring was recognized in 14 out of 200 Holstein dairy cattle raised in the cattle farm of the National Institute of Animal Science in Cheonan, Republic of Korea. Skin lesions were distributed mainly over the tail base, and sacral and perineal regions. Microscopic examinations of skin scraping samples from severely affected areas revealed numerous mites of all developmental stages. Morphologically, pedicels of the mites were short and unjointed. The tarsal suckers occurred on the pedicels of all the legs in the male worm and on the first, second, and fourth pair of legs in the adult female worm. A single long seta at the tarsus of legs III and the length of legs II being about twice as long as legs IV in adult male mites were observed. Arising anterior to the inner-most spatulate seta was a short seta with an average of 26.4 +/- 5.8 microm in length. Also, the length of setae #4 on the opisthosomal lobes was relatively short. Based on these observations, the mites were identified as Choriptes texanus. Although the chorioptic mange may not influence the mortality rate in the affected farm, reports indicate that a decline in milk production can be observed. This is the first report of chorioptic infestation in a cattle farm from Korea. PMID- 19127337 TI - Antibody responses in sera of different mouse strains experimentally infected with Neodiplostomum seoulense. AB - To examine humoral immune responses in the host, we measured serum antibody levels in different strains of mice (ICR, BALB/c, and C3H) experimentally infected with Neodiplostomum seoulense. Specific IgG antibody levels were increased remarkably with little difference among 3 strains of mice infected with N. seoulense from day 7 to 35 post-infection. More target proteins of adult parasites reacted with IgG at the time when the worm recovery decreased compared with other times. More than 20 protein bands, from 14 kDa to 94 kDa in size, were separated from the crude antigen of N. seoulense adults by SDS-PAGE, and among them 26, 30, 35, 43, 54, 67, and 94 kDa proteins were the major antigenic proteins. The results suggest that significant IgG antibody responses occur against N. seoulense in mice and this may be related with expulsion of worms. PMID- 19127338 TI - Detection of Gnathostoma spinigerum third-stage larvae in snakeheads purchased from a central part of Myanmar. AB - To examine the infection status of freshwater fish with Gnathostoma spp. larvae in Myanmar, we purchased 15 snakeheads, Channa striatus, from a local market in a suburban area of Naypyidaw, the new capital city. Two larval gnathostomes were collected using an artificial digestion technique, and observed by a light microscope and a scanning electron microscope. The size of an intact larva was 2.65 mm long and 0.32 mm wide. The characteristic morphology of the larvae included the presence of a long esophagus (0.80 mm long), 2 pairs of cervical sacs (0.43 mm long), and a characteristic head bulb with 4 rows of hooklets. The number of hooklets in the 1st, 2nd, 3rd, and 4th row was 45, 48, 50, and 52, respectively. Based on these morphological characters, the larvae were identified as the advanced 3rd-stage larvae of Gnathostoma spinigerum. This is the first report of detection of G. spinigerum 3rd-stage larvae in the central part of Myanmar. Our study suggests that intake of raw meat of snakehead fish in Myanmar may result in human gnathostomiasis. PMID- 19127339 TI - Decrease of Metagonimus yokogawai endemicity along the Tamjin River basin. AB - The Tamjin River which flows from Jangheung-gun via Gangjin-gun to the South Sea was reported to be a highly endemic area of Metagonimus yokogawai infection in 1977 and 1985. However, there were no recent studies demonstrating how much change occurred in the endemicity, in terms of prevalence and worm burden, of metagonimiasis in this river basin. Thus, a small-scale epidemiological survey was carried out on some residents along the Tamjin River basin in order to determine the current status of M. yokogawai infection. A total of 48 fecal samples were collected and examined by the Kato-Katz thick smear and formalin ether sedimentation techniques. The egg positive rate of all helminths was 50.0%, and that of M. yokogawai was 37.5%, followed by C. sinensis 22.9% and G. seoi 4.2%. To obtain the adult flukes of M. yokogawai, 6 egg positive cases were treated with praziquantel 10 mg/kg in a single dose and purged with magnesium sulfate. A total of 5,225 adult flukes (average 871 specimens per person) of M. yokogawai were collected from their diarrheic stools. Compared with the data reported in 1977 and 1985, the individual worm burdens appeared to have decreased remarkably, although the prevalence did not decrease at all. It is suggested that the endemicity of M. yokogawai infection along the Tamjin River has been reduced. To confirm this suggestion, the status of infection in snail and fish intermediate hosts should be investigated. PMID- 19127340 TI - Malondialdehyde, glutathione, and nitric oxide levels in Toxoplasma gondii seropositive patients. AB - The aim of this study was to investigate the difference in the serum malondialdehyde (MDA), glutathione (GSH), and nitric oxide (NO) levels between normal and T. gondii-infected patients. To this end, MDA, GSH, and NO levels in the sera of 37 seropositive patients and 40 participants in the control group were evaluated. In Toxoplasma ELISA, IgG results of the patient group were 1,013.0 +/- 543.8 in optical density (mean +/- SD). A statistically significant difference was found between patients and the control group in terms of MDA, GSH, and NO levels. A decrease in GSH activity was detected, while MDA and NO levels increased significantly. Consequently, it is suggested that the use of antioxidant vitamins in addition to a parasite treatment shall prove useful. The high infection vs control ratio of MDA and NO levels probably suggests the occurrence as a mechanism of tissue damage in cases of chronic toxoplasmosis. Moreover, it is recommended that the patient levels of MDA, GSH, and NO should be evaluated in toxoplasmosis. PMID- 19127341 TI - Effects of copper on T-type Ca2+ channels in mouse spermatogenic cells. AB - Low voltage-activated, rapidly inactivating T-type Ca2+ channels are found in a variety of cells, where they regulate electrical activity and Ca2+ entry. In whole-cell patch-clamp recordings from mouse spermatogenic cells, trace element copper (Cu2+) inhibited T-type Ca2+ current (IT-Ca) with IC50 of 12.06 microM. Inhibition of IT-Ca by Cu2+ was concentration-dependent and mildly voltage dependent. When voltage stepped to -20 mV, Cu2+ (10 microM) inhibited IT-Ca by 49.6 +/- 4.1%. Inhibition of IT-Ca by Cu2+ was accompanied by a shift of -2.23 mV in the voltage dependence of steady-state inactivation. Cu2+ upshifted the current-voltage (I-V) curve. To know the change of the gating kinetics of T-type Ca2+ channels, we analyzed the effect of Cu2+ on activation, inactivation, deactivation and reactivation of T-type Ca2+ channels. Since T-type Ca2+ channels are a key component in capacitation and the acrosome reaction, our data suggest that Cu2+ can affect male reproductive function through T-type Ca2+ channels as a preconception contraceptive material. PMID- 19127343 TI - Construction of a dual-tag system for gene expression, protein affinity purification and fusion protein processing. AB - An E. coli vector system was constructed which allows the expression of fusion genes via a L: -rhamnose-inducible promotor. The corresponding fusion proteins consist of the maltose-binding protein and a His-tag sequence for affinity purification, the Saccharomyces cerevisiae Smt3 protein for protein processing by proteolytic cleavage and the protein of interest. The Smt3 gene was codon optimized for expression in E. coli. In a second rhamnose-inducible vector, the S. cerevisiae Ulp1 protease gene for processing Smt3 fusion proteins was fused in the same way to maltose-binding protein and His-tag sequence but without the Smt3 gene. The enhanced green fluorescent protein (eGFP) was used as reporter and protein of interest. Both fusion proteins (MalE-6xHis-Smt3-eGFP and MalE-6xHis Ulp1) were efficiently produced in E. coli and separately purified by amylose resin. After proteolytic cleavage the products were applied to a Ni-NTA column to remove protease and tags. Pure eGFP protein was obtained in the flow-through of the column in a yield of around 35% of the crude cell extract. PMID- 19127342 TI - Diazepam administration after prolonged status epilepticus reduces neurodegeneration in the amygdala but not in the hippocampus during epileptogenesis. AB - An episode of status epilepticus (SE), if left untreated, can lead to death, or brain damage with long-term neurological consequences, including the development of epilepsy. The most common first-line treatment of SE is administration of benzodiazepines (BZs). However, the efficacy of BZs in terminating seizures is reduced with time after the onset of SE; this is accompanied by a reduced efficacy in protecting the hippocampus against neuronal damage, and is associated with impaired function and internalization of hippocampal GABA(A) receptors. In the present study, using Fluoro-Jade C staining, we found that administration of diazepam to rats at 3 h after the onset of kainic acid-induced SE, at a dose sufficient to terminate SE, had no protective effect on the hippocampus, but produced a significant reduction in neuronal degeneration in the amygdala, piriform cortex, and endopiriform nucleus, examined on days 7-9 after SE. Thus, in contrast to the hippocampus, the amygdala and other limbic structures are responsive to neuroprotection by BZs after prolonged SE, suggesting that GABA(A) receptors are not significantly altered in these structures during SE. PMID- 19127344 TI - Characterization of the enhancing effect of protamine on the proliferative activity of hepatocyte growth factor in rat hepatocytes. AB - PURPOSE: The aim of the present study was to characterize the mechanism of the stimulatory effect of protamine on HGF activity. METHODS: The enhancing effects of protamine on the proliferative activity of HGF were investigated in vivo, in primary cultured rat hepatocytes, and in perfused rat liver. RESULTS: In alpha naphthylisothiocyanate-intoxicated rats, pretreatment with protamine increased HGF-induced autophosphorylation of the HGF receptor in liver. The maximum enhancing effect of protamine on HGF-induced DNA synthesis of hepatocytes required a 10 min-pretreatment period both in vivo and in vitro, and the stimulatory effect of protamine was not observed when it was administered simultaneously with HGF. Preperfusion of the liver with protamine for 10 min decreased the non-saturable portion of hepatic clearance for (125)I-HGF, which is mainly mediated by cell-surface heparan-sulfate proteoglycan (HSPG). Inhibition of HGF binding to heparin by protamine was confirmed using heparin-coated sepharose. This inhibition also required 10 min of pretreatment, for protamine to bind heparin. CONCLUSION: The enhancing effect of protamine on the mitogenic activity of HGF on hepatocytes requires pretreatment with protamine for a short period presumably required for its binding to cell-surface heparin, implying possible regulation of c-met autophosphorylation by HSPG. PMID- 19127345 TI - Systematic review of the ability of recovery expectations to predict outcomes in non-chronic non-specific low back pain. AB - OBJECTIVE: The aim of the current review was to determine the predictive strength of low recovery expectations for activity limitation outcomes in people with non chronic NSLBP. METHODS: A systematic review of prognostic studies was performed. Included studies took baseline measures in the non-chronic phase of NSLBP, included at least one baseline measure of recovery expectation, defined as a prediction or judgement made by the person with NSLBP regarding any aspect of prognosis, and studied a sample with at least 75% of participants with NSLBP. RESULTS: Recovery expectations measured using a time-based, specific single-item tool produced a strong prediction of work outcome. Recovery expectations measured within 3 weeks of NSLBP onset provide a strong prediction of outcome. It is not clear whether predictive strength of recovery expectations is affected by the length of time between the expectation measure and outcome measure. CONCLUSION: Recovery expectations when measured using a specific, time-based measure within the first 3 weeks of NSLBP can identify people at risk of poor outcome. PMID- 19127346 TI - Development and characterisation of a novel animal model of prostate inflammation induced chronic pelvic pain. AB - Chronic pelvic pain due to prostate inflammation is a significant clinical problem. In the current study we developed and validated an animal model of inflammation-induced pelvic pain (NIH category IIIA). 3% carrageenan was injected into the ventral prostate in SD rats. At different time points (before and after 48 h, 72 h and 1 wk of injection), radiant heat and von Frey filaments (mechanical stimuli) were applied to different pelvic areas. The escape latency (s) from radiant heat, and the bending force (g) of the filament to which the animal responded by moving were taken as measures of heat and mechanical thresholds respectively. Inflamed animals showed a significant reduction in mechanical threshold (mechanical allodynia) at 72 h and 1 wk, and a significant reduction in heat threshold (thermal hyperalgesia) in the scrotal skin, compared to sham. Morphine (5 mg/kg., i.p.) significantly reduced both heat hyperalgesia and mechanical allodynia. It is expected that this novel model will prove to be useful in studying the neurobiological mechanisms of male pelvic pain. PMID- 19127347 TI - Anti-oxidative and anti-inflammatory activities of some isolated constituents from the stem bark of Allanblackia monticola Staner L.C (Guttiferae). AB - Stem bark of Allanblackia monticola has been used in association with others plant in the Cameroonian folk medicine for the treatment of various diseases such amoebic dysentery, diarrhoea, lung infections, and skin diseases. The methylene chloride fraction, its isolated compounds like alpha-mangostin, lupeol and acid betulinic were screened for antioxidant activity using free radical scavenging method. These isolated compounds were further tested for anti-inflammatory properties using carrageenan-induced model. Methylene chloride fraction, showed concentration-dependent radical scavenging activity, by inhibiting 1,1-diphenyl-1 picryl-hydrazyl radical (DPPH) with an IC(50) value of 14.60 microg/ml. alpha Mangostin and betulinic acid (500 microg/ml), showed weak radical scavenging activity with a maximum inhibition reaching 38.07 microg/ml and 26.38 microg/ml, respectively. Betulinic acid, lupeol and alpha-mangostin (5 mg/kg and 9.37 mg/kg) showed anti-inflammatory activity with a maximum inhibition of 57.89%, 57.14% and 38.70%, respectively. Methylene chloride fraction of Allanblackia monticola and some derivatives, have antioxidant and anti-inflammatory activities. PMID- 19127348 TI - Evaluation of the anti-pyretic potential of Orthosiphon stamineus Benth standardized extract. AB - The anti-pyretic activity of a standardized methanol/water (50/50) extract of Orthosiphon stamineus Benth. (SEOS) was investigated for its effect on normal body temperature and yeast-induced pyrexia in Sprague Dawley (SD) rats. The SEOS showed no effect on normal body temperature. Doses of 500 and 1000 mg/kg body weight of SEOS significantly reduced the yeast-induced elevation in body temperature. This effect persisted up to 4 h following the administration of the extract. The anti-pyretic effect of SEOS was comparable with that of paracetamol (acetaminophen in U.S) (150 mg/kg p.o.), a standard anti-pyretic agent. HPLC study revealed that rosmarinic acid, sinensetin, eupatorin and tetramethoxyflavone were present in SEOS in the amounts of 7.58%, 0.2%, 0.34% and 0.24% respectively. The LD(50) of the extract in rats was higher than 5000 mg/kg body weight. Therefore, the present study ascertained that SEOS possesses a significant anti-pyretic activity. PMID- 19127349 TI - Leptin-induced cytosolic phospholipase A2 activation in gastric mucosal protection against ethanol cytotoxicity involves epidermal growth factor receptor transactivation. AB - A pluripotent cytokine, leptin, released locally within the mucosal tissue is an important mediator of the processes of gastric mucosal defense and repair. Here, we report that leptin protection of gastric mucosal cells against ethanol cytotoxicity requires epidermal growth factor receptor (EGFR) participation. We show that the protective effect of leptin against ethanol cytotoxicity was associated with the increased EGFR and cPLA(2) phosphorylation, and characterized by a marked increase in arachidonic acid (AA) release and prostaglandin (PGE(2)) generation. The loss in countering capacity of leptin on the ethanol-induced cytotoxicity was attained with Src kinase inhibitor, PP2, and EGFR kinase inhibitor, AG1478, as well as ERK inhibitor, PD98059. Moreover, all three agents evoked also the inhibition in leptin-induced upregulation in cPLA(2) activity, AA release, and PGE(2) generation. Furthermore, changes caused by leptin in EGFR phosphorylation and cPLA(2) activation were susceptible to suppression by GM6001, a metalloprotease inhibitor of membrane-anchored EGFR ligand cleavage. These findings disclose an important link between leptin-induced and Src kinase mediated EGFR transactivation and the activation of cytosolic phospholipase A(2) that leads to up-regulation in PGE2 production, thus providing new insights into the mechanism of gastric mucosal protection by leptin. PMID- 19127350 TI - Potential of Azadirachta indica against Salmonella typhimurium-induced inflammation in BALB/c mice. AB - OBJECTIVE: To evaluate the potential of Azadirachta indica leaf extracts against Salmonella typhimurium-induced inflammation in BALB/c mice. DESIGN: Qualitative tests of A. indica leaf extracts were conducted for screening of various phytochemicals. The antiinflammatory potential of A. indica leaf extracts on S. typhimurium and its outer membrane proteins (OMPs)-induced inflammation was assessed by hyperalgesic (flicking) response of the mice inflamed paws. The monokines (IL-1alpha, IL-6 and TNF-alpha) activities in the culture supernatants of macrophages (infected with bacteria and interacted with OMPs) in the presence or absence of A. indica leaf extracts was assessed by ELISA. RESULTS: Aqueous and petroleum ether A. indica leaf extracts reduced the inflammation caused by S. typhimurium and its OMPs as assessed by paw flicking response. Petroleum ether A. indica leaf extract was found to be more effective than aqueous A. indica leaf extract. Significantly lower levels of monokines (IL-6 and TNF-alpha) were also observed in the presence of petroleum ether A. indica leaf extracts than aqueous A. indica leaf extract. These observations may be due to the presence of steroids and triterpenoids observed in petroleum ether extract. CONCLUSION: Petroleum ether A. indica leaf extract seems promising to combat S. typhimurium-induced inflammation. PMID- 19127351 TI - Effects of selenium and topiramate on lipid peroxidation and antioxidant vitamin levels in blood of pentylentetrazol-induced epileptic rats. AB - Free radicals and selenium (Se) deficiency are involved in pathogenesis of epilepsy. Topiramate (TPM), a new anticonvulsant, was reported to possess neuroprotective effect via inhibition of free radicals. We investigated the effects of Se and TPM on pentylentetrazol (PTZ)-induced blood toxicity in rats. Forty male Wistar rats were equally divided into five groups. First and second groups were used as control and PTZ group, respectively. TPM and Se were administrated to rats constituting third and forth groups for 7 days, respectively. The TPM and Se combination were given to animals in fifth group for 7 days. At the end of 7 days all groups except the first group received single dose PTZ. The brain cortex samples were taken at 3 h of PTZ administration. PTZ resulted in significant increase in plasma and erythrocytes lipid peroxidation (LP) levels although plasma vitamin E concentrations and erythrocytes glutathione peroxidase (GSH-Px) activities were reduced by PTZ. The plasma and erythrocytes LP levels in third, fourth, and fifth groups were decreased as compared to second group although GSH-Px and reduced glutathione values increased in the groups. Vitamin C and E concentrations were increased through fourth and fifth group only. Vitamin A concentrations were not changed by PTZ. In conclusion, Se and TPM seem to have protective effects on the PTZ-induced blood toxicity by inhibiting free radical supporting antioxidant redox system. PMID- 19127353 TI - Impact of training of traditional birth attendants on the newborn care. AB - OBJECTIVE: To study the impact of training of Traditional Birth Attendants (TBAs) on the Newborn care in resource poor setting in rural area. METHODS: A community based study in the Primary Health Center (PHC) area was conducted over one year period between March 2006 to February 2007. The study participants were 50 Traditional Birth Attendants (TBAs)who conduct home deliveries in the PHC area. Training was conducted for two days which included topics on techniques of conducting safe delivery and newborn care practices. Pre-test evaluation regarding knowledge and practices about newborn care was done. Post-test evaluation was done at first month (early) and at fifth month (late) after the training. Analysis was done by using Mc. Nemer's test, Chi- square test with Yates's correction and Fischer's exact test. RESULTS: Pre-test evaluation showed that, knowledge and practices about newborn care services provided by the previously trained TBAs and untrained TBAs were poor. Early and late post-test evaluation showed that, there was a progressive improvement in the newborn care provided by both the groups. Preintervention period (one year prior to the training) and postintervention period (one year after the training) showed that, there was a statistically significant (p<0.05) reduction in the perinatal deaths (11 to 3) and neonatal deaths (10 to 2) among the deliveries conducted by TBAs after the training. CONCLUSION: Training programme for TBAs with regular reinforcements in the resource poor setting will not only improve the quality of newborn care but also reduces perinatal deaths. PMID- 19127354 TI - Dental microwear patterns of extant and extinct Muridae (Rodentia, Mammalia): ecological implications. AB - Extant species of Muridae occupy a wide array of habitats and have diverse dietary habits. Consequently, their dental microwear patterns represent a potential clue to better understand the paleoecology of their extinct relatives, which are abundant in many Old World Neogene localities. In this study, dental microwear is investigated for specimens of 17 extant species of murine and deomyine rodents in order to test the reliability of this method and infer dietary preferences on the fossil species Saidomys afarensis. This extinct form comes from a mid-Pliocene site (AL 327) located at the Hadar Formation (Ethiopia) known to have delivered many hominid specimens of Australopithecus afarensis. A significant correlation between microwear patterns and diet is detected. Thus, grass, fruit, and insect eaters display, respectively, high amounts of fine scratches, wide scratches, and large pits. Moreover, some aspects of the paleoecology of S. afarensis, including feeding habits, could be assessed in regard to its dental microwear pattern. Indeed, it probably had feeding habits similar to that of living grass eaters. These results concur with the presence of open to woodland areas covered by an herbaceous vegetal layer, including monocotyledons, in the vicinity of this mid-Pliocene locality. PMID- 19127355 TI - Novel manikin for chest re-opening simulation training. PMID- 19127356 TI - Systemic and microcirculatory responses to progressive hemorrhage. AB - OBJECTIVE: To compare systemic hemodynamics with microcirculatory changes at different vascular beds during progressive hemorrhage. SETTING: University-based research laboratory. SUBJECTS: Twelve anesthetized, mechanically ventilated sheep. INTERVENTIONS: Sheep were randomly assigned to HEMORRHAGE or CONTROL group. In the HEMORRHAGE group (n = 8), three stepwise bleedings of 5 ml/kg at 30 min intervals were performed to add up 15 ml/kg. In the CONTROL group (n = 4), sheep had the same surgical preparation but were not bled. MEASUREMENTS AND MAIN RESULTS: Progressive bleeding decreased cardiac output, and superior mesenteric artery blood flow, and systemic and intestinal oxygen transports from the first step of bleeding whereas systemic and intestinal oxygen consumption remained unchanged. Mean arterial blood pressure, arterial pH and base excess, and intramucosal-arterial PCO(2) were only significantly modified in the last step of bleeding. Arterial lactate increased and sublingual, and intestinal serosal and mucosal capillary microvascular flow indexes and red blood cell velocities progressively decreased after the first step of bleeding (3.0 +/- 0.1 vs. 2.3 +/- 0.4, 3.2 +/- 0.2 vs. 2.4 +/- 0.6, 3.0 +/- 0.0 vs. 2.0 +/- 0.2, and 1,082 +/- 29 vs. 977 +/- 79, 1,042 +/- 24 vs. 953 +/- 60, 287 +/- 65 vs. 262 +/- 16 mum/s; P < 0.05 for all). CONCLUSIONS: Alterations in sublingual, intestinal microcirculation, and arterial lactate simultaneously arose from the first step of bleeding. The microcirculatory changes were identified either by semi quantitative flow index or by quantitative red blood cell velocity measurements. PMID- 19127357 TI - Actions of kurtoxin on tetrodotoxin-sensitive voltage-gated Na+ currents, NaV1.6, in murine vas deferens myocytes. AB - Kurtoxin is described as a selective inhibitor of Ca(V)3.1. Using patch-clamp techniques, the modulatory effects of kurtoxin on tetrodotoxin-sensitive voltage gated Na(+) currents (I(Na)) recorded from mouse vas deferens myocytes were investigated. Kurtoxin increased the peak amplitude of I(Na) between -40 and -30 mV, whilst inhibited the peak amplitude at more positive potentials than -10 mV, thereby demonstrating a dual action on the peak amplitude of I(Na). The time to reach the peak amplitude of I(Na) became significantly longer in the presence of kurtoxin in comparison with that of the controls. Kurtoxin also slowed the deactivation of I(Na) at more positive membrane potentials than -30 mV. Kurtoxin enhanced the total amount of electrical charge of I(Na) in a concentration dependent manner. These results suggest that kurtoxin is a modulator of Na(V)1.6 in native freshly dispersed smooth muscle cells from mouse vas deferens as well as its action on Ca(V)3.1. PMID- 19127359 TI - Transport of biomolecules in asymmetric nanofilter arrays. AB - We propose a theoretical model for describing the electric-field-driven migration of rod-like biomolecules in nanofilters comprising a periodic array of shallow passages connecting deep wells. The electrophoretic migration of the biomolecules is modeled as transport of point-sized Brownian particles, with the orientational degree of freedom captured by an entropy term. Using appropriate projections, the formulation dimensionality is reduced to one physical dimension, requiring minimal computation and making it ideal for device design and optimization. Our formulation is used to assess the effect of slanted well walls on the energy landscape and resulting molecule mobility. Using this approach, we show that asymmetry in the well shape, such as a well with one slanted and one vertical wall, may be used for separation using low-frequency alternating-current fields because the mobility of a biomolecule is different in the two directions of travel. Our results show that, compared to methods using direct-current fields, the proposed method remains effective at higher field strengths and can achieve comparable separation using a significantly shorter device. PMID- 19127360 TI - Effects of e-mails containing ADR information and a current case report on ADR reporting rate and quality of reports. AB - BACKGROUND: Spontaneous reporting of adverse drug reactions (ADRs) is one approach to increasing our knowledge on the risks of drugs in clinical practice. Under-reporting is a shortcoming of this system. OBJECTIVES: To evaluate if repeated e-mails with attachments on ADR information can affect (1) the reporting of ADRs and (2) the quality of the ADR reports. METHODS: All 117 heads of primary health care units in the region of Vastra Gotaland in Sweden were randomly allocated (1:1) to an intervention group or a control group. The intervention consisted of e-mails with attachments sent out to each of the 117 heads in January, May and September 2007. These e-mails included (1) the heading "Every ADR report is important", (2) a current case report of an ADR and (3) instructions on how to report. The number of reports from each primary health care unit run by the same head was registered, as was the quality of the report. The quality was defined as high if the ADR was (1) serious, (2) unexpected or (3) related to the use of new drugs and not labeled as common in the summary of product characteristics. All other reports were regarded as low-quality reports. RESULTS: The total number of reports increased from 89 in 2006 to 111 in 2007 (P = 0.037). No difference in the number of reports between intervention (n = 56) and control (n = 55) units could be detected. The proportion of high-quality reports before and after the intervention was 36 and 48%, respectively (intervention, P = 0.11) and 40 and 36%, respectively (control, P = 0.55). CONCLUSIONS: No apparent effect of repeated ADR e-mails on the reporting of ADRs could be detected, although an increase in the reporting rate in general was noted. PMID- 19127361 TI - Sentinel node mapping and skip metastases in patients with early gastric cancer. AB - OBJECTIVES: This study was designed to identify the characteristics of patients with early gastric cancers that have skip metastases. BACKGROUND: The possibility of lymph node metastasis is the most important factor to consider when deciding on the resection procedure for patients with early gastric cancer. METHODS: From February 2003 through July 2008, 739 patients with early gastric adenocarcinoma underwent gastric resection at the National Cancer Center, Korea, and were included in this study. Patients with skip metastases were analyzed and compared with those without skip metastases. RESULTS: Skip metastases were found in 2.8% of patients with early gastric cancer. Tumor size and the presence of lymphatic invasion were associated with skip metastases by both univariate and multivariate analysis. All skip metastases were metastases to the extraperigastric lymph nodes that skipped across the perigastric lymph nodes. Sixteen patients (66.7%) with these metastases had metastatic lymph nodes at No. 7, 8, and 9 stations. CONCLUSIONS: Tumor size should be considered during sentinel lymph node mapping to prevent false-negative results in patients with early gastric cancer. If sentinel nodes are not found in the perigastric lymph nodes, No. 7, 8, and 9 stations should be explored for prevention of false-negative sentinel node mapping results. PMID- 19127362 TI - Assessment tools in the training of surgical oncologists: educating trainees and educators. PMID- 19127363 TI - Early onset of acute lymphoblastic leukemia with MLL rearrangement after autologous stem cell transplantation for multiple myeloma. PMID- 19127364 TI - Role of frequency and mechanical index in ultrasonic-enhanced chemotherapy in rats. AB - PURPOSE: The therapeutic effect of ultrasound and micellar-encapsulated doxorubicin was studied in vivo using a tumor-bearing rat model with emphasis on how tumor growth rate is affected by ultrasonic parameters such as frequency and intensity. METHODS: This study employed ultrasound of two different frequencies (20, 476 kHz) and two pulse intensities, but identical mechanical indices and temporal average intensities. Ultrasound was applied weekly for 15 min to one of two bilateral leg tumors (DHD/K12/TRb colorectal epithelial cell line) in the rat model immediately after intravenous injection of micelle-encapsulated doxorubicin. This therapy was applied weekly for 6 weeks. RESULTS: Results showed that tumors treated with drug and ultrasound displayed, on average, slower growth rates than non-insonated tumors (P = 0.0047). However, comparison between tumors that received 20 or 476-kHz ultrasound treatments showed no statistical difference (P = 0.9275) in tumor growth rate. CONCLUSION: Application of ultrasound in combination with drug therapy was effective in reducing tumor growth rate, irrespective of which frequency was employed. PMID- 19127365 TI - Single center prospective study of tacrolimus efficacy and safety in treatment of rheumatoid arthritis. AB - The aim of this study was to prospectively evaluate the efficacy and safety of tacrolimus for treating rheumatoid arthritis (RA) patients in clinical practice. Fifty-five active RA patients who had been resistant or intolerant to other disease-modifying antirheumatic drugs were enrolled in this open-label trial. Patients were administered tacrolimus at a dosage of 1, 2 or 3 mg once daily, and followed up for 24 weeks. They were divided into three groups according to their dosage. Efficacy and safety were evaluated utilizing clinical and laboratory findings. Eighty percent of the patients had moderate or high disease activity; 55% were elderly and 53% had complications; 65% of the patients were started on tacrolimus as a monotherapy. Moderate or good response rates were achieved as follows: 38.2% (4 weeks); 41.8% (12 weeks); and 45.6% (24 weeks). Adverse events were observed in seven cases (12.7%). Only one case required hospitalization due to severe hyperglycemia caused by a high tacrolimus concentration (24.2 ng/ml); we suspected a drug interaction in this subject. Mean concentrations were dose dependent in the 1, 2, and 3 mg/day groups (2.96, 4.29, and 8.32 ng/ml, respectively). Four cases of high concentration (over 10 ng/ml), without any signs or symptoms, were observed in the 3 mg/day group; in these cases, doses were decreased and no severe adverse events occurred. Tacrolimus was found to be both effective and safe in treating active RA patients with complicated backgrounds in clinical practice. Blood concentration measurements and dose adjustments should be performed to prevent severe adverse events in a 3 mg/day group. PMID- 19127366 TI - Effect of meal size on excess post-exercise oxygen consumption in fishes with different locomotive and digestive performance. AB - Effects of feeding on pre-exercise VO(2) and excess post-exercise oxygen consumption (EPOC) after exhaustive exercise were investigated in sedentary southern catfish, active herbivorous grass carp, omnivorous crucian carp, and sluggish omnivorous darkbarbel catfish to test whether feeding had different effects on EPOC and to compare EPOC in fishes with different ecological habits. For fasting fish, the pre-exercise and peak post-exercise VO(2) were higher and recovery rates were faster in crucian carp and grass carp compared to those of darkbarbel catfish and southern catfish. EPOC magnitudes of grass carp and southern catfish were significantly larger than those of crucian carp and darkbarbel catfish. Feeding had no significant effect on peak post-exercise VO(2), recovery rate, and EPOC magnitude in grass carp. Both the pre-exercise and peak post-exercise VO(2) increased with meal size, while the EPOC magnitude and duration decreased significantly in the larger meal size groups of crucian carp and southern catfish. In darkbarbel catfish, both the pre-exercise and peak post exercise VO(2) increased with meal size, but the VO(2) increment elicited by exercise was larger in feeding groups compared with the fasting group. These results suggest that (1) the characteristics of the post-exercise VO(2) profile, such as peak post-exercise VO(2) and recovery rate, were closely related to the activity of fishes, whereas the EPOC magnitude was not and (2) the effects of feeding on EPOC were more closely related to the postprandial increase in VO(2). PMID- 19127367 TI - Food utilisation and digestive ability of aquatic and semi-terrestrial crayfishes, Cherax destructor and Engaeus sericatus (Astacidae, Parastacidae). AB - Both Engaeus sericatus and Cherax destructor are omnivorous crayfishes consuming a variety of food items. Materials identified in the faeces of both E. sericatus and C. destructor consisted of mainly plant material with minor amounts of arthropod animals, algae and fungi. The morphology of the gastric mill of C. destructor suggests that it is mainly involved in crushing of food material while the gastric mill of E. sericatus appears to be better suited to cutting of food material. Given this, the gastric mill of E. sericatus may be better able to cut the cellulose and hemicellulose fibres associated with fibrous plant material. In contrast, the gastric mill of C. destructor appears to be more efficient in grinding soft materials such as animal protein and algae. Both species accumulated high amounts of lipids in their midgut glands (about 60% of the dry mass) which were dominated by triacylglycerols (81-82% of total lipids). The dominating fatty acids were 16:0, 16:1(n-7), 18:1(n-9), 18:2(n-6), and 18:3(n-3). The two latter fatty acids can only be synthesised by plants, and are thus indicative of the consumption of terrestrial plants by the crayfishes. The similarity analysis of the fatty acid patterns showed three distinct clusters of plants and each of the crayfish species. The complement of digestive enzymes, proteinases, total cellulase, endo-beta-1,4-glucanase, beta-glucosidase, laminarinase and xylanase within midgut gland suggests that both C. destructor and E. sericatus are capable of hydrolysing a variety of substrates associated with an omnivorous diet. Higher activities of total cellulase, endo-beta-1,4 glucanase and beta-glucosidase indicate that E. sericatus is better able to hydrolyse cellulose within plant material than C. destructor. In contrast to E. sericatus, higher total protease and N-acetyl-beta-D-glucosaminidase activity in the midgut gland of C. destructor suggests that this species is better able to digest animal materials in the form of arthropods. Differences in total cellulase and gastric mill morphology suggest that E. sericatus is more efficient at digesting plant material than C. destructor. However, the contents of faecal pellets and the fatty acid compositions seem to indicate that both species opportunistically feed on the most abundant and easily accessible food items. PMID- 19127368 TI - Laparoscopic apical mesh excision for deep dyspareunia caused by mesh banding in the vaginal apex. AB - BACKGROUND: Gynecare Prolift has been successfully used for pelvic floor repair with favorable objective and subjective outcomes. There have been, however, increasing reports of significant rates of postoperative dyspareunia and impairment of sexual function. MATERIALS AND METHODS: We are presenting two cases of post Prolift dyspareunia. The patients underwent several vaginal revisions with excision of the apical bands. However, they returned soon with the same complaint. Ultimately, laparoscopic excision of the apical mesh was performed. RESULTS: Patients reported significant improvement in their intercourse pain after the procedure. CONCLUSION: Our experience shows that new onset dyspareunia is primarily caused by the apical component of the Prolift system with deep penetration pain reproduced by pressure on the fixed nondistensible apex. This problem can be treated laparoscopically with excision of the apical mesh. PMID- 19127369 TI - Heterotopic pregnancy following in vitro fertilization and embryo transfer: 12 cases report. AB - OBJECTIVE: To evaluate the incidence, predisposing factors, early diagnosis and treatment options of heterotopic pregnancy (HP) following in vitro fertilization and embryo transfer (IVF-ET) procedure. METHODS: A retrospective review study was performed to identify the HP cases after IVF-ET at the Reproductive Centre in Guangdong Women and Children's Hospital in China between the years of 2002-2007. RESULTS: Twelve out of 1,476 pregnancies (0.81%) were diagnosed for HP, of which nine patients elected for exploratory salpingectomy, two patients received selective fetal reduction by embryo aspiration under ultrasound guidance, and one patient opted for expectant treatment. Postoperatively, four intrauterine pregnancies were continued to develop until term while two were delivered at 35 weeks of gestation. The achievement ratio of continuous pregnancy was 66.7% (6/9). CONCLUSION: The incidence of HP is increasing due to the widespread use of assisted reproductive technology. An early transvaginal sonography performed by experienced radiologist/radiographer is considered to be essential and beneficial in establishing early diagnosis of HP. Both salpingectomy and selective fetal reduction by embryo aspiration can be administered as one of the effective therapies for HP with the optimal outcome of intrauterine pregnancy. PMID- 19127370 TI - Strategies to facilitate transgene expression in Chlamydomonas reinhardtii. AB - The unicellular green alga Chlamydomonas reinhardtii has been identified as a promising organism for the production of recombinant proteins. While during the last years important improvements have been developed for the production of proteins within the chloroplast, the expression levels of transgenes from the nuclear genome were too low to be of biotechnological importance. In this study, we integrated endogenous intronic sequences into the expression cassette to enhance the expression of transgenes in the nucleus. The insertion of one or more copies of intron sequences from the Chlamydomonas RBCS2 gene resulted in increased expression levels of a Renilla-luciferase gene used as a reporter. Although any of the three RBCS2 introns alone had a positive effect on expression, their integration in their physiological number and order created an over-proportional stimulating effect observed in all transformants. The secretion of the luciferase protein into the medium was achieved by using the export sequence of the Chlamydomonas ARS2 gene in a cell wall deficient strain and Renilla-luciferase could be successfully concentrated with the help of attached C terminal protein tags. Similarly, a codon adapted gene variant for human erythropoietin (crEpo) was expressed as a protein of commercial relevance. Extracellular erythropoietin produced in Chlamydomonas showed a molecular mass of 33 kDa probably resulting from post-translational modifications. Both, the increased expression levels of transgenes by integration of introns and the isolation of recombinant proteins from the culture medium are important steps towards an extended biotechnological use of this alga. PMID- 19127371 TI - Conserved inserts in the Hsp60 (GroEL) and Hsp70 (DnaK) proteins are essential for cellular growth. AB - The Hsp60 and Hsp70 chaperones contain a number of conserved inserts that are restricted to particular phyla of bacteria. A one aa insert in the E. coli GroEL and a 21-23 insert in the DnaK proteins are specific for most Gram-negative bacteria. Two other inserts in DnaK are limited to certain groups of proteobacteria. The requirement of these inserts for cellular growth was examined by carrying out complementation studies with temperature-sensitive (T(s)) mutants of E. coli groEL or dnaK. Our results demonstrate that deletion or most changes in these inserts completely abolished the complementation ability of the mutant proteins. Studies with GroEL and DnaK from some other species that either lacked or contained these inserts also indicated that these inserts are essential for growth of E. coli. The DnaK from some bacteria contains a two aa insert that is not found in E. coli. Introduction of this insert into the E. coli DnaK also led to its inactivation, indicating that these inserts are specific for different groups. We postulate that these conserved inserts that are localized in loop regions on protein surfaces, are involved in some ancillary functions that are essential for the groups of bacteria where they are found. PMID- 19127372 TI - Spontaneous spinal epidural hematoma as the initial presentation of leukemia. AB - We present a case of a 55-year-old male with progressive neurological deficits that appeared dramatically. MRI detected a spinal epidural hematoma at the cervicothoracic junction and blood tests showed leukocytosis, mild anemia, and thrombocytosis. Spontaneous spinal epidural hematoma (SSEH) as the initial presentation of leukemia was diagnosed. Urgent posterior decompression was performed after 28 h from acute onset of backache, and the patient experienced remarkable improvement in neurological findings. PMID- 19127374 TI - Comparison of radiographic changes after ACDF versus Bryan disc arthroplasty in single and bi-level cases. AB - The object of this study is to compare radiographic outcomes of anterior cervical decompression and fusion (ACDF) versus cervical disc replacement using the Bryan Cervical Disc Prosthesis (Medtronic Sofamor Danek, Memphis, TN) in terms of range of motion (ROM), Functional spinal unit (FSU), overall sagittal alignment (C2 C7), anterior intervertebral height (AIH), posterior intervertebral height (PIH) and radiographic changes at the implanted and adjacent levels. The study consisted of 105 patients. A total of 63 Bryan disc were placed in 51 patients. A single level procedure was performed in 39 patients and a two-level procedure in the other 12. Fifty-four patients underwent ACDF, 26 single level cases and 28 double level cases. The Bryan group had a mean follow-up 19 months (12-38). Mean follow-up for the ACDF group was 20 months (12-40 months). All patients were evaluated using static and dynamic cervical spine radiographs as well as MR imaging. All patients underwent anterior cervical discectomy followed by autogenous bone graft with plate (or implantation of a cage) or the Bryan artificial disc prosthesis. Clinical evaluation included the visual analogue scale (VAS), and neck disability index (NDI). Radiographic evaluation included static and dynamic flexion-extension radiographs using the computer software (Infinitt PiviewSTAR 5051) program. ROM, disc space angle, intervertebral height were measured at the operative site and adjacent levels. FSU and overall sagittal alignment (C2-C7) were also measured pre-operatively, postoperatively and at final follow-up. Radiological change was analyzed using chi(2) test (95% confidence interval). Other data were analyzed using the mixed model (SAS enterprises guide 4.1 versions). There was clinical improvement within each group in terms of VAS and NDI scores from pre-op to final follow-up but not significantly between the two groups for both single (VAS p=0.8371, NDI p=0.2872) and double (VAS p=0.2938, NDI p=0.6753) level surgeries. Overall, ROM and intervertebral height was relatively well maintained during the follow-up in the Bryan group compared to ACDF. Regardless of the number of levels operated on, significant differences were noted for overall ROM of the cervical spine (p<0.0001) and all other levels except at the upper adjacent level for single level surgeries (p=0.2872). Statistically significant (p<0.0001 and p=0.0172) differences in the trend of intervertebral height measurements between the two groups were noted at all levels except for the AIH of single level surgeries at the upper (p=0.1264) and lower (p=0.7598) adjacent levels as well as PIH for double level surgeries at the upper (p=0.8363) adjacent level. Radiological change was 3.5 times more observed for the ACDF group. Clinical status of both groups, regardless of the number of levels, showed improvement. Although clinical outcomes between the two groups were not significantly different at final follow up, radiographic parameters, namely ROM and intervertebral heights at the operated site, some adjacent levels as well as FSU and overall sagittal alignment of the cervical spine were relatively well maintained in Bryan group compared to ACDF group. We surmise that to a certain degree, the maintenance of these parameters could contribute to reduce development of adjacent level change. Noteworthy is that radiographic change was 3.5 times more observed for ACDF surgeries. A longer period of evaluation is needed, to see if all these radiographic changes will translate to symptomatic adjacent level disease. PMID- 19127375 TI - Theoretical investigation of tautomerism in N-hydroxy amidines. AB - A DFT study with QST3 approach method is used to calculate kinetic, thermodynamic, spectral and structural data of tautomers and transition state structures of some N-hydroxy amidines. All tautomers and transition states are optimized at the B3LYP/6-311++g** and B3LYP/aug-cc-pvtz level, with good agreement in energetic result with energies obtained from CBS-QB3, a complete basis set composite energy method. The result shows that the tautomer a (amide oxime) is more stable than the tautomer b (imino hydroxylamine) as is reported in the literature. In addition, our finding shows that, the energy difference between two tautomers is only in about 4-10 kcal/mol but the barrier energy found in traversing each tautomer to another one is in the range of 33-71 kcal/mol. Therefore, it is impossible to convert these two tautomers to each other at room temperature. Additionally, transition state theory is applied to estimate the barrier energy and reaction rate constants of the hydrogen exchange between tautomers in presence of 1-3 molecules of water. The computed activation barrier shows us that the barrier energy of solvent assisted tautomerism is about 9-20 kcal/mol and lower than simple tautomerism and this water-assisted tautomerism is much faster than simple tautomerism, especially with the assisting two molecules of water. PMID- 19127373 TI - Spinal extradural angiolipoma: report of two cases and review of the literature. AB - Spinal angiolipomas are benign uncommon neoplasm composed of mature lipocytes admixed with abnormal blood vessels. They account for only 0.04-1.2% of all spinal tumors. We report two cases of lumbar extradural angiolipoma and review previously reported cases. We found 118 cases of spinal epidural angiolipoma (70 females and 48 males; age range 1.5-85 years, mean 44.03) spanning from 1890 to 2006. Prior to diagnosis 40.6% of the patients had weakness of the lower limbs. The interval between the initial symptoms and tumor diagnosis ranged from 1 day to 17 years (mean 20.2 months). Except for four cases diagnosed at autopsy, 109 patients underwent surgery and gross-total resection was performed in 79 cases (72.4%). Spinal angiolipomas are tumors containing angiomatous and lipomatous tissue, predominantly located in the mid-thoracic region. All angiolipomas show iso- or hyperintensity on T1-weighted images and hyperintensity on T2-weighted images and most lesions enhance with gadolinium administration. The treatment for spinal extradural angiolipomas is total surgical resection and no adjuvant therapy should be administered. PMID- 19127377 TI - Development, malaria and adaptation to climate change: a case study from India. AB - India has reasons to be concerned about climate change. Over 650 million people depend on climate-sensitive sectors, such as rain-fed agriculture and forestry, for livelihood and over 973 million people are exposed to vector borne malarial parasites. Projection of climatic factors indicates a wider exposure to malaria for the Indian population in the future. If precautionary measures are not taken and development processes are not managed properly some developmental activities, such as hydro-electric dams and irrigation canal systems, may also exacerbate breeding grounds for malaria. This article integrates climate change and developmental variables in articulating a framework for integrated impact assessment and adaptation responses, with malaria incidence in India as a case study. The climate change variables include temperature, rainfall, humidity, extreme events, and other secondary variables. Development variables are income levels, institutional mechanisms to implement preventive measures, infrastructure development that could promote malarial breeding grounds, and other policies. The case study indicates that sustainable development variables may sometimes reduce the adverse impacts on the system due to climate change alone, while it may sometimes also exacerbate these impacts if the development variables are not managed well and therefore they produce a negative impact on the system. The study concludes that well crafted and well managed developmental policies could result in enhanced resilience of communities and systems, and lower health impacts due to climate change. PMID- 19127378 TI - Intrahepatic bilioenteric anastomosis after biliary complications of liver transplantation: operative rescue of surgical failures. AB - BACKGROUND: Biliary complications after orthotopic liver transplantation (OLT) are multifactorial in origin. In most series, the frequency of such complications ranges from 5-20%. Most can be treated by endoscopy and/or interventional radiology. For cases in which this option is not successful, surgical approach is indicated. We report the results of reoperation using an intrahepatic bilioenteric anastomosis. METHODS: The medical charts of patients with biliary complications after OLT during a 10-year period (1997-2007), who failed to respond to nonsurgical treatment and were surgically treated, were reviewed. Roux en-Y hepatojejunostomy was performed. Segments IV and V were partially removed after cutting the hilar plate, thus obtaining healthy ducts without ischemic or inflammatory reaction and allowing a wide hepatojejunostomy. RESULTS: Five cases (8.4%) with biliary complications after duct-to-duct anastomosis not amenable to further endoscopic management or interventional radiology were identified. Hepaticojejunostomy was achieved in all cases (wide, tension-free, nonischemic, fine hydrolyzable sutures), and segments IV and V were partially removed. No cholangitis, jaundice, and liver function test abnormalities were present in the postoperative. Mean follow-up was 24 months. Only one patient died of causes not related to bile duct reconstruction during follow-up. CONCLUSIONS: Intrahepatic hepatojejunostomy with partial resection of segments IV and V offers an excellent therapeutic alternative for biliary complications that require a surgical approach after OLT. PMID- 19127379 TI - Tracheobronchial lesions following esophagectomy: erosions, ulcers, and fistulae, and the predictive value of lymph node-related factors. AB - BACKGROUND: Following esophagectomy, tracheobronchial lesions (TBLs) can occur as a result of ischemia caused by extensive dissection around the tracheobronchus. In this study we assessed the causes and clinical features of these complications, paying particular attention to lymph node (LN)-related factors. METHODS: Between January 2000 and March 2007, 305 consecutive patients underwent subtotal esophagectomy using a transthoracic approach with LN dissection for thoracic esophageal cancer. TBLs, including erosions, ulcers, and fistulae, without traumatic injury during the operation, were detected during bronchoscopic examinations performed twice daily after the operation. The correlation between TBLs and tumor or surgical factors were analyzed. RESULTS: TBLs were observed in 14 patients, accounting for an overall incidence of 5%; these included 6 fistulae, 5 ulcers, and 3 erosions. Cases with TBLs significantly more often involved three-field LN dissections (3FLD) than those without TBLs. Six (43%) patients with TBLs had more than four metastatic lymph nodes, while 9 (64%) had cervical and upper-mediastinal LN metastasis (p=0.034 and 0.041, respectively). More than 60 LNs were dissected from 10 (71%) patients with TBLs (p=0.021), and logistic regression analysis revealed that dissection of more than 60 lymph nodes and 3FLD were independent predictors of TBLs. CONCLUSIONS: Esophageal cancer patients requiring extensive LN dissection of more than 60 nodes and/or 3FLD have an increased risk of developing a TBL during their postoperative course. PMID- 19127380 TI - Carotid artery stenting versus carotid endarterectomy: systematic review and meta analysis. AB - BACKGROUND: The aim of this study was to evaluate and investigate the complications of carotid endarterectomy (CEA) and carotid artery stenting (CAS) by performing a meta-analysis based on prospective randomized controlled trials (RCTs). METHODS: We performed a search of multiple electronic databases for RCTs containing patients with carotid stenosis who underwent CAS or CEA, focusing on studies published during 1995-2008. RESULTS: Eight trials with 2942 patients (1462 with CEA, 1480 with CAS) were analyzed. The pooled relative risk (RR) after CEA for stroke/death 30 days or 1 year was similar to that for CAS. Thirty-day RR = 0.69, 95% confidence interval (CI) = 0.45-1.07, p = 0.10. One-year RR = 0.88, 95% CI = 0.43-1.79, p = 0.72. The rates of death, disabling stroke, and nondisabling stroke at 30 days did not differ significantly between CEA and CAS in the subgroup analysis. Compared with CEA, the relative risk of disabling stroke/death within 30 days was not significantly less for CAS with embolic protection devices (EPDs). The relative risk of myocardial infarction within 30 days, myocardial infarction within 1 year, and cervical/peripheral nerve injury within 30 days were significantly higher after CEA; the relative risk of bradycardia/hypotension within 30 days and the 1-year restenosis rate were significantly higher after CAS. CONCLUSIONS: CAS is equal to CEA with regard to the incidence of stroke/death. These procedures may be considered complementary rather than competing modes of therapy, each of which can be optimized with careful patient selection. CAS with an EPD may be appropriate in certain patients, and in general CAS should be considered cautiously in symptomatic patients. PMID- 19127381 TI - Early indicators of treatment success after percutaneous radiofrequency of pulmonary tumors. AB - We retrospectively reviewed the imaging of patients after radiofrequency ablation (RFA) of lung metastases performed at our institution to assess the usefulness of ground glass opacification (GGO) margin for the prediction of complete tumor ablation. From January 2004 to March 2007, patients were identified where there was a postprocedure thin collimation scan to allow multiplanar reformatting, either immediately or at 24 h and at least 6 months of imaging follow-up. Thirty six tumors in 22 patients were identified. The scans were assessed for the presence and width of GGO margin, and minimal and maximal dimensions were measured. A second reviewer, blinded to the outcome of the postprocedure assessment, reviewed the follow-up imaging for recurrence. The recurrence group had larger tumors (p = 0.045) and smaller mean minimal GGO margin width (p = 0.0001). Multivariate binary regression analysis confirmed that the minimal GGO margin was significantly (p < 0.005) associated with tumor recurrence. Receiver operator characteristic curve analysis suggests a cutoff of 4.5 mm for complete tumor ablation. There was substantial agreement (kappa = 0.759) between the site of absent GGO margin and the site of tumor recurrence. The point on the tumor surface where there is no GGO margin is likely to be the site of future recurrence. In our experience, a circumferential GGO margin of >5 mm is the minimal margion required to ensure complete tumor ablation. PMID- 19127382 TI - HU participates in expression of a specific set of genes required for growth and survival at acidic pH in Escherichia coli. AB - The major histone-like Escherichia coli protein, HU, is composed of alpha and beta subunits respectively encoded by hupA and hupB in Escherichia coli. A mutant deficient in both hupA and hupB grew at a slightly slower rate than the wild type at pH 7.5. Growth of the mutant diminished with a decrease in pH, and no growth was observed at pH 4.6. Mutants of either hupA or hupB grew at all pH levels tested. The arginine-dependent survival at pH 2.5 was diminished approximately 60 fold by the deletion of both hupA and hupB, whereas the survival was slightly affected by the deletion of either hupA or hupB. The mRNA levels of adiA and adiC, which respectively encode arginine decarboxylase and arginine/agmatine antiporter, were low in the mutant deficient in both hupA and hupB. The deletion of both hupA and hupB had little effect on survival at pH 2.5 in the presence of glutamate or lysine, and expression of the genes for glutamate and lysine decarboxylases was not impaired by the deletion of the HU genes. These results suggest that HU regulates expression of the specific set of genes required for growth and survival in acidic environments. PMID- 19127383 TI - Effects of nasal CPAP treatment on insulin resistance, lipid profile, and plasma leptin in sleep apnea. AB - BACKGROUND: Obstructive sleep apnea has been linked with metabolic syndrome characterized by dyslipidemia, dyscoagulation, hypertension, and diabetes mellitus type 2 and their cardiovascular consequences. This study was designed to determine the effects of 8 weeks of therapy with continuous positive airway pressure (CPAP) on insulin resistance, glucose, and lipid profile, and the relationship between leptin and insulin-resistance parameters in patients with moderate-to-severe obstructive sleep apnea. METHODS: In 44 patients, serum cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, very low-density lipoprotein, leptin, and insulin parameters were measured at baseline and after 8 weeks of CPAP. Insulin resistance index was based on the homeostasis model assessment (HOMA-IR) method. Insulin sensitivity (HOMA-S) and insulin secretion capacity (HOMA-beta) also were calculated. Thirteen patients were excluded from statistical analyses due to noncompliant CPAP usage (<4 h night(-1)). RESULTS: In 31 patients who used CPAP for > or =4 h night(-1), CPAP therapy reduced total cholesterol (P < 0.05), low-density lipoprotein (P < 0.05), and leptin (P < 0.05). Circulating leptin levels showed significant correlation with both HOMA-S and HOMA-IR at baseline and follow-up (P = 0.03 for all). In addition, there was no correlation between HOMA-IR and the severity of sleep apnea, which was shown by apnea-hypopnea index. CONCLUSIONS: In patients with moderate-to-severe obstructive sleep apnea, compliant CPAP usage may improve insulin secretion capacity, reduce leptin, total cholesterol, and low-density lipoprotein levels. Leptin showed significant relationship with insulin resistance, and this relationship remained after 8 weeks of CPAP therapy. PMID- 19127384 TI - Response of the egg parasitoids Trissolcus basalis and Telenomus podisi to compounds from defensive secretions of stink bugs. AB - We tested the hypotheses that host-searching behavior of the egg parasitoids Telenomus podisi and Trissolcus basalis may be differentially influenced by the different blends of volatiles released from the metathoracic glands of adult stink bug host species. We further studied whether such a differential response is due to different individual components of these glands and whether these responses reflect host preferences. Y-tube olfactometer bioassays were carried out with crude extracts of metathoracic glands of five different host species of neotropical stink bugs. Additionally, we tested the parasitoids' responses to synthetic standards of individual compounds identified in these stink bug glands. Results showed that females of T. basalis and T. podisi responded differentially to crude gland extracts of the different species of host stink bugs and to the compounds tested. The parasitoid T. basalis showed a positive taxic behavior to Nezara viridula methathoracxic gland extracts of a host species preferred in the field, i.e., N. viridula. Furthermore, T. basalis responded positively to 4-oxo (E)-2-hexenal and (E)-2-decenal, two components of N. viridula glandular secretion. Higher residence time, reduced linear velocity, and higher tortuosity in the arm of the olfactometer supplied with 4-oxo-(E)-2-hexenal showed that this compound modifies the kinetics of some traits of T. basalis walking pattern and suggests that it might stimulate the searching behavior of this parasitoid. The parasitoid T. podisi was attracted to crude gland extracts of the preferred host (Euschistus heros) and also to 4-oxo-(E)-2-hexenal. Additionally, this parasitoid responded positively to (E)-2-hexenal and to the hydrocarbon tridecane, both of which are defensive compounds released from the metathoracic glands by several stink bugs. The results indicate some degree of specialization in the response of two generalist parasitoid species toward defensive secretions of stink bugs. PMID- 19127385 TI - The chlorophyll catabolite, pheophorbide a, confers predation resistance in a larval tortoise beetle shield defense. AB - Larval insect herbivores feeding externally on leaves are vulnerable to numerous and varied enemies. Larvae of the Neotropical herbivore, Chelymorpha alternans (Chrysomelidae:Cassidinae), possess shields made of cast skins and feces, which can be aimed and waved at attacking enemies. Prior work with C. alternans feeding on Merremia umbellata (Convolvulaceae) showed that shields offered protection from generalist predators, and polar compounds were implicated. This study used a ubiquitous ant predator, Azteca lacrymosa, in field bioassays to determine the chemical constitution of the defense. We confirmed that intact shields do protect larvae and that methanol-water leaching significantly reduced shield effectiveness. Liquid chromatography-mass spectrometry (LC-MS) of the methanolic shield extract revealed two peaks at 20.18 min and 21.97 min, both with a molecular ion at m/z 593.4, and a strong UV absorption around 409 nm, suggesting a porphyrin-type compound. LC-MS analysis of a commercial standard confirmed pheophorbide a (Pha) identity. C. alternans shields contained more than 100 microg Pha per shield. Shields leached with methanol-water did not deter ants. Methanol-water-leached shields enhanced with 3 microg of Pha were more deterrent than larvae with solvent-leached shields, while those with 5 microg additional Pha provided slightly less deterrence than larvae with intact shields. Solvent leached shields with 10 microg added Pha were comparable to intact shields, even though the Pha concentration was less than 10% of its natural concentration. Our findings are the first to assign an ecological role for a chlorophyll catabolite as a deterrent in an insect defense. PMID- 19127386 TI - The association between the receipt of lipid lowering therapy and HIV status among veterans who met NCEP/ATP III criteria for the receipt of lipid lowering medication. AB - OBJECTIVE: To examine the association between HIV infection status and the receipt of lipid lowering therapy based on National Cholesterol Education Program/Adult Treatment Panel (NCEP/ATP III) guidelines and to assess whether HIV viral load and hepatitis C (HCV) status alters that association. PARTICIPANTS AND DESIGN: A cross-sectional analysis of survey, laboratory, and pharmacy data from 1,577 male participants (59% HIV infected) of the Veterans Aging Cohort Five-Site Study, a prospective observational cohort of U.S. veterans with and without HIV infection. MEASUREMENTS: Receipt of lipid lowering therapy obtained from the VA pharmacy benefits management system was the main outcome. RESULTS: The prevalence of lipid lowering therapy among HIV-infected and HIV-uninfected veterans was 15.4% vs. 37.9%, respectively, p < 0.01. Among veterans who met NCEP/ATP III criteria for lipid lowering therapy, HIV-infected veterans had a significantly lower prevalence for the receipt of lipid lowering therapy (adjusted odds ratio (OR) = 0.43, 95% confidence interval (C.I.) 0.28-0.67) as compared with HIV uninfected veterans. Among HIV-infected veterans, log HIV viral load (adjusted OR = 0.57, 95% CI, 0.41-0.81) and HIV-HCV co-infection (adjusted OR = 0.31, 95% CI = 0.13-0.75) were negatively associated with receipt of lipid lowering therapy. Exposure to HAART was not associated with receipt of lipid lowering therapy. CONCLUSIONS: Among those who met NCEP/ATP III criteria for lipid lowering therapy, HIV-infected veterans, particularly those with high HIV viral loads and HCV co-infection, were significantly less likely to receive lipid lowering therapy. This may be a modifiable mediator of cardiovascular disease among HIV infected individuals. PMID- 19127387 TI - Risk-group targeted inferior vena cava filter placement in gastric bypass patients. AB - BACKGROUND: Despite a growing body of evidence guiding appropriate perioperative thromboprophylaxis in the general population, few data direct strategies to reduce deep venous thrombosis (DVT) and pulmonary embolism (PE) in the morbidly obese. We have implemented a novel protocol for venous thromboembolism (VTE) risk stratification in Roux-en-Y gastric bypass (RYGB) candidates at our institution, which augments clinical assessment with screening for thrombophilias, to guide retrievable inferior vena cava (IVC) filter utilization. METHODS: A retrospective review of prospectively collected data from patients who underwent primary RYGB between 2001 and 2008 at the University of North Carolina at Chapel Hill was completed. During that time, clinical assessment of VTE risk was amplified by focused plasma screening for common thrombophilias (factors VIII, IX, and XI, d dimer, fibrinogen). Preoperative prophylactic IVC filters were offered to high risk patients. The database was reviewed for perioperative DVTs, PEs, and filter related complications. RESULTS: Of 330 patients, in 162 attempts, 160 had prophylactic IVC filters placed with four complications overall (2.47%). No patient had symptoms of PE during the planned 6-week filter period, though one had a PE occur immediately after filter removal (0.63%); in contrast, five of 170 patients (2.94%) without prophylactic IVC filters presented with symptomatic PE (p = 0.216). In total, 147 (91.88%) prophylactic filters were removed. CONCLUSIONS: Risk-group targeted prophylactic inferior vena cava filter placement prior to RYGB is safe with a trend towards reduced occurrence of PE. PMID- 19127388 TI - Obstructive sleep apnea-hypopnea syndrome--the role of bariatric and maxillofacial surgeries. AB - Obstructive sleep apnea-hypopnea syndrome (OSAHS) is a complex disease with multifactorial etiology. It is marked by the occurrence of apnea and hypopnea events caused by repeated obstructions of the upper airways. OSAHS is strongly associated with obesity, and the prevalence of this disease in morbidly obese patients is very high. Nevertheless, not all patients with OSAHS are obese, and for this reason, there may be other anatomical predispositions to airway collapse. In obese patients, fatty deposition in the parapharyngeal region results in airway reduction and predisposes to airway collapse, worsened by neurologic loss of the normal dilator muscle tone of the neck. However, in nonobese patients, specific craniofacial characteristics such as posterior air pharyngeal space, tongue length, hyoid position, and maxillomandibular deficiencies may predispose some people to develop OSAHS. Treatment strategies for OSAHS patients vary from clinical treatment with continuous positive airway pressure, oral appliances, or medications for mild and moderate OSAHS patients, bariatric surgery for severe obese OSAHS patients to maxillomandibular advancement for obese or nonobese OSAHS patients. PMID- 19127390 TI - When a man thinks he has female traits--constructing femininity and masculinity: methodological potentials and limitations. AB - In everyday life, people are confronted with common beliefs about how women and men differ from each other. These beliefs make them wonder about their personal femininity and masculinity. But what hides behind these constructs? In this article, we will discuss the potential and limitations of different definitions. We will focus on methodological issues which can trigger new discussions about the social implications of gender differences-which are not part of this paper. One of the major questions here is whether it is methodologically justifiable (legitimate) to make individual predictions based on group differences. An issue that is not only relevant in regards to femininity and masculinity. In general, all definitions of masculinity and femininity show more limitations than potentials. They are either neglecting certain sub-groups or are based on stereotypes (either displaying common beliefs or exaggerated group differences). Individual predictions derived from these constructs are questionable due to oversimplification/reductionism (e.g., when forming groups). By pointing out that constructs like femininity and masculinity can in fact limit an individual's development, it is suggested to clearly differentiate between different types of differences and to clearly state what can and cannot be said when "measuring" an individual's masculinity and femininity. Since femininity and masculinity can be seen as example constructs, and generalizations are part of many research processes, general implications beyond these concepts are being discussed. PMID- 19127391 TI - Mainstream economics and sense-making. AB - This work presents a semiotic approach to the economy, underlining that any economic phenomena is at the same time a communicative act as it is contingent to sense-making. The article discusses this topic by focusing on a specific phenomenon studied by economics: the underground economy. It shows that the conceptualization of the underground economy in terms of sense-making processes offers a thought-provoking perspective for theoretical development. More in general, the discussion proposed makes it clear that in order to deepen our vision of economic phenomena in a more thoughtful and realistic way we need to rethink these phenomena as being reciprocally and circularly embedded in the semiotic flow of life. The economy is within sense-making and it is shaped by it; at the same time sense-making is within the economy, as its semiotic substance. PMID- 19127392 TI - 50 years ago in CORR: Slipping of the upper femoral epiphysis. Beckett Howorth MD. CORR 1957;10:148-173. PMID- 19127394 TI - Magnetic resonance angiography in clubfoot and vertical talus: a feasibility study. AB - Congenital vascular alterations of the normal adult arterial pattern have been associated with multiple congenital limb deformities including clubfoot and vertical talus. Investigators have observed absence of the anterior tibial artery and dorsalis pedis artery in most patients with clubfoot, and absence of the posterior tibial artery in all patients with vertical talus. We used magnetic resonance angiography to define the lower extremity vascular anatomy of two patients with left-sided vertical talus and right-sided clubfoot and one patient with bilateral vertical talus and cartilage-derived morphogenetic protein-1 (CDMP 1) gene mutation. Of the three patients, one had bilateral posterior tibial artery deficiencies while the other had bilateral anterior tibial artery deficiencies. The third patient with bilateral vertical talus and CDMP-1 mutation had normal arterial structure bilaterally. Though clubfoot and vertical talus have distinctly different clinical phenotypes, the association of each with arterial abnormalities suggests a common etiology during development. The presence of normal arterial structure in our patient with vertical talus and CDMP 1 mutation suggests that other nonvascular etiologies may be responsible for some cases of foot deformities. LEVEL OF EVIDENCE: Level IV, prognostic case series. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19127395 TI - Combining CT and nuclear: a winning hybrid team. PMID- 19127396 TI - Combination of anatomic and perfusion imaging for decision making in a professional soccer player with giant coronary artery to left ventricle fistula. PMID- 19127410 TI - Application of genome-wide SNP data for uncovering pairwise relationships and quantitative trait loci. AB - The genetic analysis of quantitative traits in humans is changing as a result of the availability of whole-genome SNP data. Heritability analysis can make use of actual genetic sharing between pairs of individuals estimated from the genotype data, rather than the expected genetic sharing implied by their family relationship. This could provide more accurate heritability estimates and help to overcome the equal environment assumption. Quantitative trait locus (QTL) linkage mapping can make use of local genetic sharing inferred from very dense local genotype data from pedigree members or individuals not previously known to be related. This approach may be particularly suited for detecting loci that contain rare variants with major effect on the phenotype. Finally, whole-genome SNP data can be used to measure the genetic similarity between individuals to provide matched sets for association studies, in order to avoid spurious association from population stratification. PMID- 19127411 TI - A PEX10 defect in a patient with no detectable defect in peroxisome assembly or metabolism in cultured fibroblasts. AB - Zellweger spectrum disorders (ZSD) are diagnosed by biochemical assay in blood, urine and cultured fibroblasts and PEX gene mutation identification. In most cases studies in fibroblasts corroborate results obtained in body fluids. In 1996 Clayton and colleagues described a 10-year old girl with evidence of a peroxisome disorder, based on elevated bile acid metabolites and phytanate. At the time it was not possible to distinguish whether she had a ZSD or a single peroxisomal protein defect. Studies in our laboratory showed that she also had elevated plasma pipecolate, supporting the former diagnosis. Despite the abnormal metabolites detected in blood (phytanate, bile acid intermediates and pipecolate), analysis of multiple peroxisomal pathways in fibroblasts yielded normal results. In addition, she had a milder clinical phenotype than usually associated with ZSD. Since complementation analysis to determine the gene defect was not possible, we screened this patient following the PEX Gene Screen algorithm (PGS). The PGS provides a template for sequencing PEX gene exons independent of complementation analysis. Two mutations in PEX10 were identified, a frameshift mutation inherited from her father and a de novo missense mutation in a conserved functional domain on the other allele. This case highlights that molecular analysis may be essential to the diagnosis of patients at the milder end of the ZSD spectrum. Furthermore, it supports the concept that some tissues are less affected by certain PEX gene defects than brain and liver. PMID- 19127412 TI - Role of Foxa1 in regulation of bcl2 expression during oxidative-stress-induced apoptosis in A549 type II pneumocytes. AB - Forkhead box protein A1 (Foxa1) is an evolutionarily conserved winged helix transcription factor that was traditionally considered to be involved in embryonic development and cell differentiation. However, little is known about the role of Foxa1 in oxidative-stress-induced apoptosis. In this study, hydrogen peroxide (H(2)O(2))-induced apoptosis, upregulation of Foxa1, and the role of Foxa1 in the regulation of bcl2 gene expression were studied in A549 type II pneumocytes. H(2)O(2) upregulated Foxa1 mRNA and protein in a time- and dose dependent manner. Overexpression of Foxa1 promoted apoptosis, whereas Foxa1 deficiency, induced by antisense oligonucleotides, decreased A549 cell apoptosis induced by H(2)O(2), as shown by flow cytometry. Moreover, Foxa1 overexpression decreased the expression of bcl2, while Foxa1 depletion increased the expression of bcl2. Electrophoretic mobility shift assay and chromatin immunoprecipitation revealed that Foxa1 bound to bcl2 promoter, and H(2)O(2) promoted its DNA binding activity. Luciferase reporter showed that Foxa1 also decreased the transcription activity of bcl2 promoter under normal conditions and oxidative stress. These results indicate that Foxa1 plays a pro-apoptotic role by inhibiting the expression of anti-apoptotic gene bcl2. PMID- 19127413 TI - A qualitative study examining Latino functional health literacy levels and sources of health information. AB - The purpose of this study was to determine the health information sources used by Latinos in southwest Ohio, identify individual Latino residents' functional health literacy levels, and identify any access barriers to those sources of health information. Results show almost two-thirds had low acculturation levels to US culture. Overall, the major source of health information is a medical setting, followed by media technology (which included the Internet). However, when it comes to being ill, the primary source becomes a media choice, then medical. The barriers to accessing health information included language and lack of confidence/knowledge. Participants reported moderate satisfaction with the sources of health information available, and had an 'adequate' health literacy level in Spanish. This study was important because it filled an existing information gap for the Latino community, a racial ethnic minority population in the southwest Ohio area. With the results of this study, health educators and other health care practitioners might be better able to understand the health care needs of the Latino community and could essentially create improved and culturally competent health communications. PMID- 19127414 TI - Preferences among immigrant Hispanic women for written educational materials regarding upper respiratory infections. AB - The need for culturally appropriate health education materials for Hispanic populations has been widely recognized, and Spanish-language materials are available through a number of private and governmental organizations. We convened two focus groups to elucidate preferences regarding how health-related messages are obtained and to identify which educational materials available in Spanish were preferred by 26 recently immigrated Hispanic homemakers who had received 15 different bimonthly written documents as part of a community-based clinical trial to prevent household transmission of colds and influenza. Participants gave three primary reasons for volunteering to participate in the study: to provide better care for their children (96.2%, 25/26), to get information (96.2%, 25/26), and to get free products (47.1%, 8/17). Their primary sources of health-related information were relatives and friends (42.9%, 6/14), clinicians (35.7%, 5/14), mass media (14.3%, 2/14) or the emergency room (7.1%, 1/14); none mentioned the internet. Materials using either a question and answer or true/false format were clearly preferred, even when other options were more colorful or had lower reading levels. Printed educational materials may be ineffective unless they include a more systematic assessment of the user's perceived needs for the information as well as consideration of format. In this population, a question and answer or true/false format and materials that could be shared with their children were greatly preferred. PMID- 19127415 TI - Recruiting the future public health workforce: an analysis of prospect communication among accredited Schools of Public Health. AB - Schools of Public Health cannot sustain the national momentum for public health justice and human rights without recruiting and training a skilled public health workforce. With growing demand for public health workers, schools must work to increase their applicant pools. This project examined prospect communication materials from accredited Schools of Public Health and found that the vast majority of schools did not capitalize on opportunities to move prospects to applicants. Whereas most responded within a reasonable time, several schools made no communication efforts at all. Recruitment materials varied widely from institution to institution and between epidemiology and health education programs. Strategic, personalized communication strategies-the 3 Cs-are recommended to increase the pools of qualified applicants nationwide and can be utilized to increase prospect pools in a wide range of health sciences. PMID- 19127416 TI - Evaluation of a hepatitis B lay health worker intervention for Chinese Americans and Canadians. AB - Hepatitis B testing is recommended for immigrants from countries where hepatitis B infection is endemic. However, only about one-half of Chinese in North America have received hepatitis B testing. We conducted a randomized controlled trial to evaluate the effectiveness of a hepatitis B lay health worker intervention for Chinese Americans/Canadians. Four hundred and sixty individuals who had never been tested for hepatitis B were identified from community-based surveys of Chinese conducted in Seattle, Washington, and Vancouver, British Columbia. These individuals were randomly assigned to receive a hepatitis B lay health worker intervention or a direct mailing of physical activity educational materials. Follow-up surveys were completed 6 months after randomization. Self-reported hepatitis B testing was verified through medical records review. A total of 319 individuals responded to the follow-up survey (69% response rate). Medical records data verified hepatitis B testing since randomization for 9 (6%) of the 142 experimental group participants and 3 (2%) of the 177 control group participants (P = 0.04). At follow-up, a higher proportion of individuals in the experimental arm than individuals in the control arm knew that hepatitis B can be spread by razors (P < 0.001) and during sexual intercourse (P = 0.07). Our findings suggest that lay health worker interventions can impact hepatitis B related knowledge. However, our hepatitis B lay health worker intervention had a very limited impact on hepatitis B testing completion. Future research should evaluate other intervention approaches to improving hepatitis B testing rates among Chinese in North America. PMID- 19127417 TI - Evaluation of group genetic counseling for hereditary breast and ovarian cancer. AB - As demand for genetic counseling regarding hereditary cancer continues to grow, more efficient methods of providing this service must be explored. In this pilot study, group genetic counseling was offered to two different cohorts of women seeking genetic counseling for Hereditary Breast and Ovarian Cancer. Seven group sessions, designed to cover all aspects of an individual genetic counseling appointment, were conducted. Although patients were receptive to group genetic counseling, a significant proportion chose individual counseling when given the option. Advantages of group genetic counseling include shared experience and increased efficiency. Disadvantages include increased frustration at not being eligible for genetic testing, group influence on decision-making, privacy concerns, increased need for follow-up, and difficulty booking group appointments. Overall, the level of patient satisfaction with group genetic counseling was similar to that of individual counseling. The results of this pilot study suggest that further research is needed to determine whether group genetic counseling is an acceptable alternative to individual counseling. PMID- 19127418 TI - Cultural values and caregiver preference for Mexican-American and non-Latino White elders. AB - This study examined caregiver preferences in the event of hip fracture between Mexican-American and non-Latino White elders. The differential effects of ethnicity and a cultural factor were also examined to elucidate the role of culture on caregiver preference.Data came from a cross-sectional survey of 89 Mexican-American and 30 non-Latino White elders. Hierarchical binary logistic regression was used to examine the differential impact of ethnicity and a cultural factor on caregiver preference. Fewer Mexican-American elders than non Latino White elders preferred to rely on a formal/professional helper, and a greater proportion of Mexican-Americans than non-Latino Whites would turn to informal caregivers when faced with care needs following a hip fracture. The cultural factor significantly mediated the ethnic effect on caregiver preference. The need for culturally-relevant services based on caregiver preference for long term care is discussed in addressing health disparities for ethnically diverse elders. PMID- 19127419 TI - Case report: color as a therapeutic intervention. PMID- 19127420 TI - Coping with racism: a selective review of the literature and a theoretical and methodological critique. AB - Racism is a stressor that contributes to racial/ethnic disparities in mental and physical health and to variations in these outcomes within racial and ethnic minority groups. The aim of this paper is to identify and discuss key issues in the study of individual-level strategies for coping with interpersonal racism. We begin with a discussion of the ways in which racism acts as a stressor and requires the mobilization of coping resources. Next, we examine available models for describing and conceptualizing strategies for coping with racism. Third, we discuss three major forms of coping: racial identity development, social support seeking and anger suppression and expression. We examine empirical support for the role of these coping strategies in buffering the impact of racism on specific health-related outcomes, including mental health (i.e., specifically, self reported psychological distress and depressive symptoms), self-reported physical health, resting blood pressure levels, and cardiovascular reactivity to stressors. Careful examination of the effectiveness of individual-level coping strategies can guide future interventions on both the individual and community levels. PMID- 19127421 TI - Disparities in the provision of medical care: an outcome in search of an explanation. AB - This purpose of this paper is to review the literature on racial/ethnic disparities in the utilization and quality of care and the proposed explanations for these differences. First, the literature on racial/ethnic disparities in medical treatment is reviewed briefly with the goal of providing a sense of the range of procedures and conditions on which these disparities occur. Then, the possible role of physician/provider, patient, and health care system factors in contributing to these disparities is reviewed. Finally, suggestions for new or expanded directions for research in each of these three areas are given. The goal of the paper is to identify factors that might be particularly amenable to the type of research done by health psychologists. PMID- 19127422 TI - Identifying people with acute HIV infection: demographic features, risk factors, and use of health care among individuals with AHI in North Carolina. AB - Identifying and counseling individuals with Acute HIV Infection (AHI) offers a critical opportunity to avert preventable HIV transmission, however, opportunities to recognize these individuals may be missed. We surveyed 32 adults diagnosed with AHI during voluntary HIV testing from 1/1/03 to 2/28/05 in publicly funded testing sites in NC to describe their clinical, social, and behavioral characteristics. Eighty-one percent of participants were men; 59% were African American. Seventy-five percent experienced symptoms consistent with acute retroviral syndrome; although 83% sought medical care for these symptoms, only 15% were appropriately diagnosed at that initial medical visit, suggesting opportunities to diagnose these individuals earlier were missed. Eighty-five percent of the men engaged in sex with men. More than 50% of the participants thought they were infected with HIV by a steady partner. This study yields important information to assist in identifying populations at risk for or infected with AHI and designing both primary and secondary prevention interventions. PMID- 19127424 TI - Stress and coping in HIV-positive former plasma/blood donors in China: a test of cognitive appraisal theory. AB - Throughout the 1990s, many villagers in rural China were infected with HIV through commercial plasma/blood donation. These former plasma/blood donors (FPDs) experienced many HIV-related stressors. This study tested a cognitive appraisal model of stress and coping in a sample of HIV-positive adult FPDs. Participants (N = 207) from multiple villages completed a battery of questionnaires assessing HIV-related stress, HIV symptoms, cognitive appraisal, coping behaviors, and psychological distress. Participants reported high levels of HIV-related stress, depression, and anxiety. In a structural equation model, greater HIV-related stress, HIV symptoms, and threat appraisal were directly associated with psychological distress. HIV-related stress was also indirectly associated with psychological distress through threat appraisal. In a second model, coping was found to mediate the relationship between challenge appraisal and psychological distress. Results support the utility of cognitive appraisal theory. Stress management interventions targeting HIV-positive FPDs in China are indicated. PMID- 19127425 TI - Selecting measures for use in child mental health services: a scorecard approach. AB - The implementation of evidence-based treatments in mental health services requires reliable and valid measurements to guide treatment. This study evaluated the efficiency of three caretaker-report measures of child psychiatric disorders. Data from 211 caregivers were used to assess the CBCL, the OHIO, and the SDQ. A scorecard methodology was implemented to determine the efficiency of each scale as compared to the DISC-IV. Across measures, the OHIO was optimal for assessing services need while the CBCL and SDQ provided better disorder-specific assessment. Results may improve clinical practice by providing an empirical approach to the selection of assessment tools. PMID- 19127426 TI - Child welfare recommendations to improve mental health services for children who have experienced abuse and neglect: a national perspective. AB - This study uses a key informant approach to understand the nature, extent, and quality of outpatient mental health services for children in the child welfare system (CWS) in the United States. We interviewed 89 county child welfare administrators to determine the status of outpatient mental health services and provide recommendations for enhancing care and service delivery. Developed for this study (Caring for Children in Child Welfare), the interview was incorporated in the second formal data collection wave (i.e., 18 months after study baseline assessment) of the National Survey of Child and Adolescent Well-Being. The results highlighted general variability in the degree to which these agencies used evidence-based interventions within outpatient services, demonstrated clinical expertise with this population, and met the needs of their families. Community agency use of evidence-based interventions was found to predict their effectiveness in improving clients' mental health problems. Proposed suggestions for service improvement varied across domains and reflected the need for more communication/coordination, service access, options and resources, and practice refinements to accommodate families' needs. We discuss the implications of these recommendations from CWS stakeholders for enhancing the service delivery system. PMID- 19127427 TI - Comparison of letrozole with continuous gonadotropins and clomiphene-gonadotropin combination for ovulation induction in 1387 PCOS women after clomiphene citrate failure: a randomized prospective clinical trial. AB - PURPOSE: Letrozole, though reported to be an effective ovulation inducing agent, warrants larger randomized trials. The purpose of this study is to compare the efficacy of letrozole with that of rFSH and clomiphene citrate(CC)/rFSH for ovarian stimulation in IUI cycles. METHODS: Randomized, prospective, single blinded clinical trial. 1387 PCOS women after CC failure were randomized into three groups: Group A received letrozole, Group B received CC with two doses rFSH and Group C received continuous rFSH day 2 onwards until hCG injection. RESULTS: Group A, B and C had an ovulation rate of 79.30%, 56.95% and 89.89% and cycle cancellation rate of 20.70%, 43.05% and 10.11%, respectively. Pregnancy rates in Group A, B and C were 23.39%, 14.35% and 17.92%, while the miscarriage rates were 13.80%, 16.67% and 14.52%, respectively. CONCLUSION: Letrozole appears to be a suitable ovulation inducing agent in PCOS women with CC failure and is found to be most effective when baseline estradiol level >60 pg/ml. PMID- 19127428 TI - Haplotypes of XRCC1 and survival outcome in patients with metastatic breast cancer. PMID- 19127429 TI - Effects of exogenous excitatory amino acid neurotransmitters on blood-brain barrier disruption in focal cerebral ischemia. AB - This study was performed to determine whether exogenous N-methyl-D: -aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) would aggravate blood-brain barrier (BBB) disruption in focal cerebral ischemia in rats. Forty-five minutes after middle cerebral artery (MCA) occlusion, one of the following patches was applied to the exposed ischemic cerebral cortex of each rat: normal saline (control), 10(-5) M AMPA, 10(-4) M AMPA, 10(-5) M NMDA, or 10( 4) M NMDA. At 1 h after MCA occlusion, BBB permeability was determined by measuring the transfer coefficient (Ki) of (14)C-alpha-aminoisobutyric acid ((14)C-AIB). In all experimental groups, the Ki of the ischemic cortex (IC) was higher than that of the corresponding contralateral cortex (CC). The Ki of the IC of the animals treated with 10(-4) M AMPA or 10(-4) M NMDA was higher (+41%: P < 0.05 and +33%: P < 0.05, respectively) than that of the control animals. Our data demonstrated that exogenous NMDA or AMPA could further aggravate the BBB disruption in focal cerebral ischemia. Any insult increasing the release of excitatory neurotransmitters could further aggravate BBB disruption and brain edema during the ischemic period. PMID- 19127430 TI - Glial metabolism of valine. AB - The three essential amino acids, valine, leucine and isoleucine, constitute the group of branched-chain amino acids (BCAAs). BCAAs are rapidly taken up into the brain parenchyma, where they serve several distinct functions including that as fuel material in brain energy metabolism. As one function of astrocytes is considered the production of fuel molecules that support the energy metabolism of adjacent neural cells in brain. Astroglia-rich primary cultures (APC) were shown to rapidly dispose of the BCAAs, including valine, contained in the culture medium. While the metabolisms of leucine and isoleucine by APC have already been studied in detail, some aspects of valine metabolism remained to be determined. Therefore, in the present study an NMR analysis was performed to identify the (13)C-labelled metabolites that are generated by APC during catabolism of [U (13)C]valine and that are subsequently released into the incubation medium. The results presented show that APC (1) are potently disposing of the valine contained in the incubation medium; (2) are capable of degrading valine to the tricarboxylic acid (TCA) cycle member succinyl-CoA; and (3) release into the extracellular milieu valine catabolites and compounds generated from them such as [U-(13)C]2-oxoisovalerate, [U-(13)C]3-hydroxyisobutyrate, [U-(13)C]2 methylmalonate, [U-(13)C]isobutyrate, and [U-(13)C]propionate as well as several TCA cycle-dependent metabolites including lactate. PMID- 19127431 TI - Impact of the patient-provider relationship on the survival of foreign born outpatients with tuberculosis. AB - BACKGROUND: Managing tuberculosis in foreign born patients entails a complex interaction between patient and provider. METHODS: Using a retrospective cohort study and survival analysis, this study evaluates the impact of patient and provider factors on the survival of foreign born outpatients with active tuberculosis. The primary outcome of the study is 1 year all-cause mortality. RESULTS: In our cohort, patient-provider language discordance was associated with an increased risk of death [HR: 2.33; 95% CI: 1.39-3.88], while receiving treatment from a tuberculosis experienced physician [HR: 0.41; 95% CI: 0.22-0.77] and treatment in a dedicated tuberculosis clinic [HR: 0.53; 95% CI: 0.29-0.98] was associated with a lower risk of death. DISCUSSION: Patient-provider communication and health systems factors played a large role in the survival of our cohort of foreign born tuberculosis outpatients. These findings suggest that language barriers and the provision of care by experienced providers in specialized clinic settings may have important effects on health outcomes. PMID- 19127432 TI - Lessons learned from two school tuberculosis investigations. AB - We describe the results from tuberculosis (TB) contact investigations of two high school students. Following the development of active TB in two foreign-born students, contact investigations were performed to detect contacts with active TB disease or latent TB infection (LTBI). The two students developed pulmonary TB within 2 years of immigrating to the United States. Among household contacts, no case of active TB was identified; however, LTBI was identified in 7 of 20 persons screened (35%). Of the 104 high-risk school contacts identified, no cases of active TB disease were found, but 7 (9.3%) were diagnosed with LTBI. An additional 683 low-risk contacts were screened and 9 (1.5%) were positive. Schools and Public Health departments needs to be prepared for outbreak investigations and should screen only persons with a high risk of exposure to the index case with active TB in an attempt to identify secondary infections. Those persons with a low risk of exposure should not be screened. PMID- 19127433 TI - Prevalence of metabolic syndrome in US-born Latin and Caribbean youth. AB - BACKGROUND: Little is knows about the prevalence of the metabolic syndrome (MS) among overweight first or second generation United States immigrant children/adolescents from Central/South America and the Caribbean basin. METHODS: Analysis of anthropometric and laboratory data (N = 224) in overweight children ages 3-18 (64% Hispanic, 25% AfroCaribbean/black, 8% white, 3% multiracial) was conducted. The main outcome measure was the prevalence of individual parameters of MS and the MS (defined as > or =3 abnormal components). RESULTS: The prevalence of the MS was 29% for the overall sample; an additional 28% had two MS components. Boys were significantly more likely than girls to have abnormal systolic blood pressure (P < 0.05). Hispanics were significantly more likely than blacks to have abnormal triglyceride and HDL cholesterol (P < 0.01 for both comparisons). CONCLUSIONS: Our results indicate that both sub-groups of, and major ethnic groups (Mexican- and African American) are at equal risk for cardiometabolic disease. Early identification of MS in recent immigrant children may improve their future cardiometabolic health. PMID- 19127434 TI - The disconnected values (intervention) model for promoting healthy habits in religious institutions. AB - The purpose of this article is to provide an intervention model that can be used by religious leaders for changing health behavior among practicing members of religious communities. The intervention does not require extensive training or licensure in counseling psychology. At the heart of this model is the acknowledgement that a person's negative habits (e.g., lack of exercise, poor nutrition) and his or her deepest values and beliefs (e.g., faith, health, family) are often misaligned, or disconnected. In addition, the unhealthy outcomes from these habits are contrary to the scriptural traditions of the world religions and thus are especially relevant to individuals who practice their religious beliefs. The Sacred Scriptures of Judaism and Christianity, for example, are replete with teachings that extol the virtues of practicing habits that promote good health and energy. In addition, evidence is mounting in the existing health intervention literature that adopting permanent and desirable changes in health behavior have not been successful, and that adherence to desirable habits such as exercise and proper nutrition is short-lived. The Disconnected Values Model (DVM) provides a novel approach for enhancing health behavior change within the context of the mission of most religious institutions. The model is compatible with skills presented by religious leaders, who possess more credibility and influence in changing the behavior of members and service attendees of their respective religious institutions. The religious leader's role is to provide the client with faith-based incentives to initiate and maintain changes in their health behaviors, and perhaps to provide resources for the individual to pursue an action plan. A case study is described in which the DVM intervention was used successfully with an individual of strong faith. PMID- 19127435 TI - Changes in childhood immunization disparities between central cities and their respective states, 2000 versus 2006. AB - Central cities have lower childhood immunization coverage rates than states in which they are located. We conducted a secondary analysis of the National Immunization Survey (NIS) 2000 and 2006 of children 19-35 months old for 26 NIS defined central cities and the rest of their respective states in order to examine patterns in early childhood immunization disparities between central cities and their respective states and the contextual factors associated with these disparities. We examined three measures of immunization disparities (absolute, difference, and ratio of change) and the patterns of disparity change with regard to selected contextual factors derived from the census. In 2000, immunization coverage in central cities was 68.3% and 74.7% in the rest of their states, a 6.4% disparity (t = 3.82, p < 0.000). Between 2000 and 2006, the overall city/state disparity narrowed to 3.5%, with the central city coverage up to 78.7% vs. 82.5% for the rest of state (t = 2.48, p = 0.017). However, changes in immunization disparities were not uniform: six cities narrowed, 14 had minimal change, and six widened. Central cities with a larger share of Hispanics experienced less reduction in disparities than other cities (beta = -4.2, t = 2.11, p = 0.047). Despite overall progress in childhood immunization coverage, most central cities still show significant disparities with respect to the rest of their states. Cities with larger Hispanic populations may need extra help in narrowing their disparities. PMID- 19127436 TI - Indinavir-loaded pH-sensitive microparticles for taste masking: toward extemporaneous pediatric anti-HIV/AIDS liquid formulations with improved patient compliance. AB - The aim of this work was to develop indinavir pediatric anti-HIV/AIDS formulations enabling convenient dose adjustment, ease of oral administration, and improved organoleptic properties by means of the generation of drug-loaded microparticles made of a polymer that is insoluble under intake conditions and dissolves fast in the stomach in order to completely release the active agent. Indinavir-loaded microparticles made of a pH-dependent polymeric excipient soluble at pH < 5, Eudragit E100, were prepared using a double emulsion solvent diffusion technique and the in vitro release profiles characterized. Finally, taste masking properties were evaluated in blind randomized sensory experiments by ten healthy human volunteers. The use of a w/o/o emulsion system resulted in indinavir loads around 90%. Thermal analysis of the microparticles by differential scanning calorimetry revealed that indinavir appeared mainly dispersed at the molecular level. Concentrations of residual organic solvents as determined by gas chromatography were below the upper limits specified by the European Pharmacopeia for pharmaceutical oral formulations. Then, the behavior of drug-containing microparticles in aqueous media at different pH values was assessed. While they selectively dissolved in gastric-like medium, in tap water (intake conditions), the matrix remained almost unchanged and efficiently prevented drug dissolution. Finally, sensoring taste tests performed by volunteers indicated that systems with indinavir loads approximately 15% displayed acceptable taste. This work explored the production of indinavir containing microparticles based on a common pharmaceutical excipient as a means for the improvement of medicines of drugs involved in the treatment of HIV/AIDS. For systems containing about 15% drug, taste studies confirmed the acceptability of the formulation. In pediatric regimes, this composition would require an acceptable amount of formulation (0.7-1.5 g). PMID- 19127437 TI - Heralding the new publisher: strategic alliance and improved service. PMID- 19127438 TI - Increases in positive reappraisal coping during a group-based mantram intervention mediate sustained reductions in anger in HIV-positive persons. AB - BACKGROUND: There is evidence that various meditation practices reduce distress, but little is known about the mechanisms of frequently repeating a mantram-a spiritual word or phrase-on distress reduction. Mantram repetition is the portable practice of focusing attention frequently on a mantram throughout the day without a specific time, place, or posture. PURPOSE: We examined the hypothesis of whether increases in positive reappraisal coping or distancing coping mediated the sustained decreases in anger found following a group-based mantram intervention that was designed to train attention and promote awareness of internal experiences. METHOD: A secondary analysis was performed on data collected from a randomized controlled trial that compared a group-based mantram intervention (n = 46) to an attention-matched control (n = 47) in a community sample of human immunodeficiency virus-positive adults. Positive reappraisal and distancing coping were explored as potential mediators of anger reduction. RESULTS: Participants in the mantram intervention reported significant increases in positive reappraisal coping over the 5-week intervention period, whereas the control group reported decreases. Increases in positive reappraisal coping during the 5-week intervention period appear to mediate the effect of mantram on decreased anger at 22-week follow-up. CONCLUSIONS: Findings suggest that a group based mantram intervention may reduce anger by enhancing positive reappraisal coping. PMID- 19127440 TI - Risk perceptions and behaviour: towards pandemic control of emerging infectious diseases : international research on risk perception in the control of emerging infectious diseases. PMID- 19127439 TI - Willingness to take travel-related health risks--a study among Finnish tourists in Asia during the avian influenza outbreak. AB - BACKGROUND: Health risk perception and behavior of tourists during epidemics is a challenge for behavioral medicine. PURPOSE: The purpose was to analyze associations of psychosocial factors and willingness to take health risks on holiday and business trips. METHOD: Subjects (survey n = 338) were Finnish tourists visiting South-East Asia during the avian influenza epidemic of 2004. On holidays, 13.8%, and on business trips, 6.3% would take (rather) high risks, 14.1% reported having tendency to take health risks. Willingness to take health risks on both kinds of trips was lower among those +40 years old than those <40. Comparatively high risk-taking tendency and high perceived HIV risk were related to the increased willingness to take health risks on both kinds of trips. RESULTS: On holidays, willingness to take health risks was related to trust in fate, and on business trips, this was also related to trust in God and less precautionary behavior of avoiding hand shaking, but also to higher estimation of other risks in life. CONCLUSION: Younger travelers and those on holidays are willing to take more health risks than those who are older or on business trips. Travel advice during epidemics could be differentially targeted to different age groups and to holiday and business travelers. PMID- 19127441 TI - Effect of heating strategy on power consumption and performance of a pilot plant anaerobic digester. AB - The effect of heating strategy on power consumption and performance of a pilot plant anaerobic digester treating chicken litter, under thermophilic conditions, has been studied. Heating strategy was evaluated using three different spans (0.2 degrees C, 0.6 degrees C, and 1.0 degree C) for triggering the temperature control system from target temperature (56.7 degrees C). The hydraulic retention time in the pilot plant digester was in the range of 32 to 37 days, varying the total solids concentration fed from 5% to 6%. The results showed that under the experimental conditions, heating was the most energy-demanding process with 95.5% of the energy used. Increments up to 7.5% and 3.8%, respectively, on mechanical and heating power consumption, were observed as the span, for triggering the temperature control system from target temperature, was increased. Under the experimental conditions studied here, an increment of 30.6% on the global biodigester performance index was observed when a span of 1.0 degree C was compared to the one of 0.2 degrees C. PMID- 19127442 TI - Cell immobilization with polyurethane foam for retaining Trichoderma reesei cells during foam fractionation for cellulase collection. AB - In situ affinity foam fractionation is a potential powerful tool for continuous, selective removal of products from bioprocesses. When evaluating its applicability to cellulase production by Trichoderma reesei fermentation, we encountered the difficulty of significant removal of fungal mycelia along with the cellulase. To solve this problem, cell immobilization using cut pieces of hydrophilic polyurethane (PU) foam was evaluated. Five commercial PU foams with different pore sizes and porosities were tested. Two were found to support good cell growth, cellulase production, and cell loading (about 0.6 g dry cells per g PU). The PU-immobilized mycelia were successfully retained in the foaming process. PMID- 19127443 TI - Aspergillus fumigatus thermophilic and acidophilic endoglucanases. AB - This study evaluated the production of cellulolytic enzymes by an Aspergillus fumigatus strain, isolated from sugar cane bagasse, according to its ability to grow on microcrystalline cellulose as the sole carbon source. The effect of the carbon source (brewer's spent grain, sugarcane bagasse, and wheat bran) and of the nitrogen source (corn steep liquor and sodium nitrate) on cellulase production was studied using submerged and solid state cultivations at 30 degrees C. The highest levels of endoglucanase (CMCase) corresponded to 365 U L(-1) and was obtained using sugarcane bagasse (1%) and corn steep liquor (1.2%) in submerged fermentation within 6 days of cultivation. This supernatant was used to run a sodium dodecyl sulfate polyacrylamide gel electrophoresis that showed six bands with endoglucanase activity. CMCase activity was higher at 65 degrees C and pH 2.0, indicating that this microorganism produces a thermophilic and acid endoglucanase. Solid state cultivation favored FPase production, that reached 47 U g(-1) of dry substrate (wheat bran and sugarcane bagasse) within 3 days. PMID- 19127444 TI - Controlled production of fructose by an exoinulinase from Aspergillus ficuum. AB - An exoinulinase has been isolated, purified and characterised from a commercially available broth of Aspergillus ficuum. The enzyme was purified 4.2-fold in a 21% yield with a specific activity of 12,300 U mg(-1)(protein) after dialysis, ammonium sulphate fractionation and Sephacryl S-200 size exclusion and ion exchange chromatography. The molecular weight of this enzyme was estimated to be 63 kDa by SDS-PAGE. It exhibited a pH and temperature optima of 5.4 and 50 degrees C respectively and under such conditions the enzyme remained stable with 96% and 63.8% residual activity after incubation for 12 h and 72 h respectively. The respective K (m) and V (max) values were 4.75 mM and 833.3 micromol min(-1) ml(-1), respectively. Response surface methodological statistical analysis was evaluated for the maximal production of fructose from the hydrolysis of pure commercial chicory inulin. Incubation of the dialyzed crude exoinulinase (100 U/ml, 48 h, 50 degrees C, 150% inulin, pH 5.0) produced the highest amount of fructose (106.4 mg/ml) under static batch conditions. The purified exoinulinase was evaluated for fructose production and the highest amount (98 mg/ml) was produced after 12 h incubation at 50 degrees C, 150% inulin pH 5.0. The use of a crude exoinulinase preparation is economically desirable and the industrial production of fructose from inulin hydrolysis is biotechnologically feasible. PMID- 19127445 TI - Carboxylate platform: the MixAlco process part 1: comparison of three biomass conversion platforms. AB - To convert biomass to liquid fuels, three platforms are compared: thermochemical, sugar, and carboxylate. To create a common basis, each platform is fed "ideal biomass," which contains polysaccharides (68.3%) and lignin (31.7%). This ratio is typical of hardwood biomass and was selected so that when gasified and converted to hydrogen, the lignin has sufficient energy to produce ethanol from the carboxylic acids produced by the carboxylate platform. Using balanced chemical reactions, the theoretical yield and energy efficiency were determined for each platform. For all platforms, the ethanol yield can be increased by 71% to 107% by supplying external hydrogen produced from other sources (e.g., solar, wind, nuclear, fossil fuels). The alcohols can be converted to alkanes with a modest loss of energy efficiency (3 to 5 percentage points). Of the three platforms considered, the carboxylate platform has demonstrated the highest product yields. PMID- 19127446 TI - Estrogen regulation of the neprilysin gene through a hormone-responsive element. AB - Our previous data showed that neprilysin (NEP), a zinc metalloendopeptidase, which can degrade amyloid-beta peptide (Abeta) whose central nerve system accumulation is the primary cause of Alzheimer's disease (AD), responds to estrogen in the brain. Recently, it has been shown that the transcription of the neprilysin gene can be up regulated by progesterone, androgens, and glucocorticoids through two androgen response elements within the NEP gene--an androgen response region (ARR) and an androgen response element (ARE). Through a yeast report gene system, we now find that the ARR but not the ARE respond to estrogen. However, androgen could efficiently enhance the expression of the report gene mainly through ARE. Our results indicate that the decrease of NEP, caused by the decline of estrogen or androgen with aging, may be an important factor leading to Abeta accumulation and AD. PMID- 19127447 TI - Induction of adult human bone marrow mesenchymal stromal cells into functional astrocyte-like cells: potential for restorative treatment in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder with its motor phenomena due mostly to loss of dopamine-producing neurons in the substantia nigra. Pharmacological treatments aimed to increase the deficient dopaminergic neurotransmission are effective in ameliorating the cardinal symptoms, but none of these therapies is curative. It has been suggested that treatment with neurotrophic factors (NTFs) might protect and prevent death of the surviving dopaminergic neurons and induce proliferation of their axonal nerve terminals with reinnervations of the deafferented striatum. However, long-term delivery of such proteins into the CNS is problematic. We therefore aimed to differentiate ex vivo human bone marrow-derived mesenchymal stromal cells into astrocyte-like cells, capable of generating NTFs for future transplantation into basal ganglia of PD patients. Indeed, mesenchymal stromal cells treated with our novel astrocyte differentiation medium, present astrocyte-like morphology and express the astrocyte markers S100beta, glutamine synthetase and glial fibrillary acidic protein. Moreover, these astrocyte-like cells produce and secrete significant amounts of glial cell line-derived neurotrophic factor (GDNF), nerve growth factor (NGF), and brain-derived neurotrophic factor as indicated by messenger RNA, real-time polymerase chain reaction, ELISA, and Western blot analyses. Such NTF-producing cells transplanted into the striatum of 6-hydroxydopamine-lesioned rats, a model of PD, produced a progressive reduction in the apomorphine-induced contralateral rotations as well as behavioral improvement in rotor-rod and the "sunflower seeds" eating motor tests. Histological assessments revealed that the engrafted cells survived and expressed astrocyte and human markers and acted to regenerate the damaged dopaminergic nerve terminal system. Findings indicate that our novel procedure to induce NTF-producing astrocyte-like cells derived from human bone marrow stromal cells might become a promising and feasible autologous transplantation strategy for PD. PMID- 19127449 TI - Practice, politics, and papers. PMID- 19127448 TI - Monitoring of cerebrovascular autoregulation: facts, myths, and missing links. AB - The methods for continuous assessment of cerebral autoregulation using correlation, phase shift, or transmission (either in time- or frequency-domain) were introduced a decade ago. They express dynamic relationships between slow waves of transcranial Doppler (TCD), blood flow velocity (FV) and cerebral perfusion pressure (CPP), or arterial pressure (ABP). We review a methodology and clinical application of indices useful for monitoring cerebral autoregulation and pressure-reactivity in various scenarios of neuro-critical care. FACTS: Poor autoregulation and loss of pressure-reactivity are independent predictors of fatal outcome following head injury. Autoregulation is impaired by too low or too high CPP when compared to autoregulation with normal CPP (usually between 60 and 85 mmHg; and these limits are highly individual). Hemispheric asymmetry of the bi laterally assessed autoregulation has been associated with asymmetry of CT scan findings: autoregulation was found to be worse ipsilateral to contusion or lateralized edema causing midline shift. The pressure-reactivity (PRx index) correlated with a state of low CBF and CMRO2 revealed using PET studies. The PRx is easier to monitor over prolonged periods of time than the TCD-based indices as it does not require fixation of external probes. Continuous monitoring with the PRx can be used to direct CPP-oriented therapy by determining the optimal CPP for pressure-reactivity. Autoregulation indices are able to reflect transient changes of autoregulation, as seen during plateau waves of ICP. However, minute-to-minute assessment of autoregulation has a poor signal-to-noise ratio. Averaging across time (30 min) or by combining with other relevant parameters improves the accuracy. MYTHS: It is debatable whether the TCD-based indices in head injured patients can be calculated using ABP instead of CPP. Thresholds for functional and disturbed autoregulation dramatically depends on arterial tension of CO2- therefore, comparison between patients cannot be performed without comparing their PaCO2. The TCD pulsatility index cannot accurately detect the lower limit of autoregulation. MISSING LINKS: We still do not know whether autoregulation oriented therapy can be understood as a consensus between CPP-directed protocols and the Lund-concept. What are the links between endothelial function and autoregulation indices? Can autoregulation after head injury be improved with statins or EPO, as in subarachnoid hemorrhage? In conclusion, monitoring cerebral autoregulation can be used in a variety of clinical scenarios and may be helpful in delineating optimal therapeutic strategies. PMID- 19127450 TI - Primary anorectal malignant melanoma--a case report and review of literature. PMID- 19127451 TI - An update on advance directives in the medical record: findings from 1186 consecutive patients with unresectable exocrine pancreas cancer. AB - BACKGROUND: The Terri Schiavo case and other recent events underscore the importance of advance directives. Yet, in the past, only a small subgroup has utilized them. This study from a large tertiary medical center was undertaken to assess current rates of advance directives among patients with incurable pancreas cancer. METHODS/RESULTS: The medical records of 1,186 consecutive patients with unresectable pancreas cancer were reviewed over a 4-year span. Only 174 patients (15%) had an advance directive in the medical record. Older age and having cancer therapy at our institution were associated with a greater likelihood of having an advance directive with odds ratios (95% confidence intervals) of 8.26 (2.81, 24.93) and 2.86 (2.03, 4.02), respectively, in multivariate analyses. Importantly, 42 patients (24%) had a different person designated as their healthcare agent in their advanced directive than what appeared in the medical record as the "contact person." CONCLUSION: These findings underscore the ongoing need to discuss advance directives with patients with incurable malignancies and to clarify patients' wishes when seemingly contradictory information appears in other parts of the medical record. PMID- 19127453 TI - DSN depletion is a simple method to remove selected transcripts from cDNA populations. AB - A novel DSN-depletion method allows elimination of selected sequences from full length-enriched cDNA libraries. Depleted cDNA can be applied for subsequent EST sequencing, expression cloning, and functional screening approaches. The method employs specific features of the kamchatka crab duplex-specific nuclease (DSN). This thermostable enzyme is specific for double-stranded (ds) DNA, and is thus used for selective degradation of ds DNA in complex nucleic acids. DSN depletion is performed prior to library cloning, and includes the following steps: target cDNA is mixed with excess driver DNA (representing fragments of the genes to be eliminated), denatured, and allowed to hybridize. During hybridization, driver molecules form hybrids with the target sequences, leading to their removal from the ss DNA fraction. Next, the ds DNA fraction is hydrolyzed by DSN, and the ss fraction is amplified using long-distance PCR. DSN depletion has been tested in model experiments. PMID- 19127454 TI - HLA-DQB1* polymorphism and associations with dilated cardiomyopathy, inflammatory dilated cardiomyopathy and myocarditis. AB - To date, only weak associations have been reported between idiopathic dilated cardiomyopathy (DCM) and certain HLA class II alleles. Associations between HLA class II alleles and specific causes of DCM, especially myocarditis, have as yet not been systematically investigated. Typing of HLA DQB1* allele was performed using a sequence-specific primer-polymerase chain reaction technique in 22 unrelated patients with idiopathic DCM, 19 patients with myocarditis and normal left ventricular function, and 16 patients with myocarditis and impaired left ventricular function (i.e. inflammatory DCM). Controls comprised 44 patients without (inflammatory) cardiac disease and a population control. A significant association was found for presence of HLA DQB1*0303 with myocarditis without cardiac dysfunction. Weaker associations were seen for presence of HLA DQB1*0301 and absence of HLA DQB1*06 with inflammatory DCM. Additionally, allelic combination DQB1*02-DQB1*03 may be able to distinguish idiopathic from inflammatory DCM, and HLA DQB1*02 myocarditis with cardiac dysfunction from myocarditis without, if results are confirmed by larger prospective studies. PMID- 19127455 TI - Gene polymorphisms of TNF-alpha-308 (G/A), IL-10(-1082) (G/A), IL-6(-174) (G/C) and IL-1Ra (VNTR) in Egyptian cases with type 1 diabetes mellitus. AB - BACKGROUND: Type 1 diabetes (T1D) is a genetically conditioned autoimmune disease in which cytokines play an important role. Objectives. To check for the association of polymorphisms of cytokine genes with type 1 diabetes. Subjects. This work included 50 cases with T1D and 98 healthy individuals from the Nile Delta region of Egypt. Cases included 20 males and 30 females with a median age of 25 and range of 15-50 years. METHODS: DNA was amplified using PCR with sequence-specific primers for detection of polymorphisms related to tumor necrosis factor (TNF)-alpha(- 308) (G/A), interleukin (IL)-10(- 1082) (G/A), IL 6(- 174) (G/C), and IL-1Ra (VNTR). RESULTS: Cases with T1D showed significant higher frequency of genotypes of TNF-alpha(- 308) AA (p < 0.001, odds ratio (OR) = 7.91), IL-6-17CC (p < 0.05, OR = 3.36) and IL-1Ra A1A1 (p < 0.05, OR = 3.68) with significant lower frequencies of TNF-alpha(- 308) GA, and IL-1Ra A1A2 genotypes (p < 0.001 and < 0.05, respectively). They also showed significant higher frequency of TNF-alpha(- 308) allele A (p < 0.05, OR = 2.0), IL-1Ra allele A1 (p < 0.05, OR = 2.98) with a significant lower frequency of TNF-alpha(- 308) G allele and IL-1Ra A2 allele (p < 0.05). No significant difference was detected among cases in relation to IL-10(- 1082) (G/A) genotypes or alleles nor in relation to age, sex, consanguinity or family history of the disease. CONCLUSIONS: Polymorphisms related to TNF-alpha and IL-1Ra genes may be considered genetic markers for T1D among Egyptians with a potential impact on family counseling and management. PMID- 19127456 TI - BioPlex 2200 multiplexed system: simultaneous detection of anti-dsDNA and anti chromatin antibodies in patients with systemic lupus erythematosus. AB - BACKGROUND: Antibodies for double-stranded DNA (anti-dsDNA) and chromatin represent specific markers of systemic lupus erythematosus (SLE). AIMS: (1) To evaluate the analytical performance of a multiplexed bead assay (BioPlex 2200) for the simultaneous detection of anti-dsDNA and anti-chromatin antibodies, (2) to compare the results for anti-dsDNA with those obtained using Farr assay, and (3) to analyze the clinical relevance of these antibodies when applied to the follow-up of SLE patients with active nephritis. PATIENTS AND METHODS: Hundred and five clinically characterized SLE patients and 96 healthy blood donors sera were analyzed by BioPlex 2200. RESULTS: Prevalence of these antibodies was significantly higher (p < 0.0001) in SLE patients than in controls (68 and 70% for anti-dsDNA and anti-chromatin, vs. 1% for both anti-dsDNA and anti-chromatin, respectively). If you consider a sample positive if either anti-dsDNA and/or anti chromatin is positive, then the prevalence of these antibodies reached 78% (82/105) in SLE patients. For anti-dsDNA measurements, the kappa coefficient was 0.59 between BioPlex 2200 and Farr assay. Comparison between SLE patients with and without nephritis in a follow-up study showed that patients with active nephritis were associated with an increase of anti-dsDNA and anti-chromatin levels and a reduction of CH50, whereas no variation of antibody levels was observed in SLE patients without nephritis. CONCLUSION: Our results demonstrated a benefit of simultaneously measuring anti-dsDNA and anti-chromatin in SLE patients. The BioPlex 2200 achieved good analytical performances and proved to be a useful method for monitoring and diagnosing SLE. PMID- 19127457 TI - TH17 (and TH1) signatures of intestinal biopsies of CD patients in response to gliadin. AB - Celiac disease (CD) is an immunological disorder caused by intolerance to ingested gliadin and other cereal prolamins that has been included in the T(H)1 dominated group of diseases, where IL-12 induced IFNgamma is the major proinflamatory signal. Recently, another linage of T cells has been described, namely T(H)17, characterized by production of IL-17, that differentiate in response to TGFbeta and IL-6 and participate in the pathogenesis of several autoimmune diseases. Using RT-PCR analysis of gene expression, we analyzed the presence of T(H)1 (IL-12 and IFNgamma) and T(H)17 (TGFbeta, IL-6, IL-17A, IL-17F and IL-23) related cytokines in intestinal biopsies from CD patients with active disease compared to remission and from treated patients after acute, in vitro re exposure to gliadin. Potent T(H)1 and T(H)17 responses were present in the active stage of the disease, whereas short incubation of normalized biopsies with gliadin did not increase the expression of the effector cytokines, although a tendency of upregulation for both T(H)1 and T(H)17 promoting factors was observed, suggestive of a reactivation of proinflammatory pathways. These results place CD into the group of autoimmune disorders in which T(H)17 cells also participate, although the relative importance of each T cell response and their role in the initial events of the disease need further investigation. PMID- 19127458 TI - Increased percentages of autoantibodies in immunoglobulin A-deficient children do not correlate with clinical manifestations. AB - IgA deficiency (IgAD) is frequently associated with autoimmune phenomena. The aim of this study is to evaluate the frequency of 22 different autoantibodies in 60 patients with IgAD and to examine the physical and other laboratory findings of the suspected cases for autoimmune diseases. The evaluated autoantibodies were Anti-nuclear antibody (ANA) profile (autoantibodies against RNP/Sm, SS-A, Ro-52, SS-B, Scl-70, Pm-Scl, Jo-1, centromere B, PCNA, dsDNA, nucleosomes, histones, ribozomal P-protein, AMA-M2), anti-cardiolipin IgG and IgM, anti-neutrophilic cytoplasmic antibodies (ANCA), rheumatoid factor (RF), anti-thyroglobulin (anti T) and anti-thyroid microsomal antigen (anti-M) and direct cooms test. Forty-one healthy children were included as a control group. ANA titers < or = 1:80 were accepted as normal and titers > or = 1:80 are accepted as positive. In ANA screening, 14 patients showed positivity in different titres. Seven of them were equal to or below 1:80. The other seven patients (11.6%) had positive ANA titers (>1:160) whereas three of them had anti-dsDNA, anti-histon and anti-centromer antibodies. These patients did not have any clinical and laboratory signs of autoimmune diseases. ANA positivity was found higher in IgA deficient children (p < 0.05) compared to controls. RF and pANCA were found positive during follow-up of two different selective IgAD patients. IgG and IgM antibodies against cardiolipin, direct coombs, anti-T and anti-M tests were not found positive in any subjects. In conclusion, increased frequency of autoantibodies in IgAD patients may often be observed. However, the detection of autoantibodies do not show or predict whether this patient will develop an autoimmune disease. PMID- 19127459 TI - Olfaction, psychiatric disorders and autoimmunity: is there a common genetic association? AB - Psychiatric diseases are often associated with mild alterations in immune functions (e.g., schizophrenia) as well as autoimmune features. Recent evidence suggests that autoimmune diseases (AD) demonstrate a higher prevalence of psychiatric disorders, such as depression and psychosis, than in the normal population. Patients with AD often have an olfactory impairment as well, based on smell studies, accompanied by olfactory regional alterations in neuroimaging. Some evidence suggests that olfactory gene receptors have additional functions in the brain, related to their direct anatomical connection to the limbic system. For example, odor sensing may explain HLA-dissimilar mate selection in humans and animals. Recently, a large cluster of the olfactory receptor (OR) genes was mapped in proximity to the HLA locus on chromosome 6. The HLA and linked OR genes are clustered in haplotypes and are highly polymorphic. This finding may constitute an association among autoimmunity, psychiatric disorders and smell impairment. In this review, we examine the anatomic, genetic and clinical clues that may support an association among these conditions. PMID- 19127460 TI - Some controversial multiple testing problems in regulatory applications. AB - Multiple testing problems in regulatory applications are often more challenging than the problems of handling a set of mathematical symbols representing multiple null hypotheses under testing. In the union-intersection setting, it is important to define a family of null hypotheses relevant to the clinical questions at issue. The distinction between primary endpoint and secondary endpoint needs to be considered properly in different clinical applications. Without proper consideration, the widely used sequential gate keeping strategies often impose too many logical restrictions to make sense, particularly to deal with the problem of testing multiple doses and multiple endpoints, the problem of testing a composite endpoint and its component endpoints, and the problem of testing superiority and noninferiority in the presence of multiple endpoints. Partitioning the null hypotheses involved in closed testing into clinical relevant orderings or sets can be a viable alternative to resolving the illogical problems requiring more attention from clinical trialists in defining the clinical hypotheses or clinical question(s) at the design stage. In the intersection-union setting there is little room for alleviating the stringency of the requirement that each endpoint must meet the same intended alpha level, unless the parameter space under the null hypothesis can be substantially restricted. Such restriction often requires insurmountable justification and usually cannot be supported by the internal data. Thus, a possible remedial approach to alleviate the possible conservatism as a result of this requirement is a group-sequential design strategy that starts with a conservative sample size planning and then utilizes an alpha spending function to possibly reach the conclusion early. PMID- 19127466 TI - Use of Bayesian methods for multivariate bioequivalence measures. AB - In this paper, we introduce a Bayesian analysis for bioequivalence data assuming multivariate pharmacokinetic measures. With the introduction of correlation parameters between the pharmacokinetic measures or between the random effects in the bioequivalence models, we observe a good improvement in the bioequivalence results. These results are of great practical interest since they can yield higher accuracy and reliability for the bioequivalence tests, usually assumed by regulatory offices. An example is introduced to illustrate the proposed methodology by comparing the usual univariate bioequivalence methods with multivariate bioequivalence. We also consider some usual existing discrimination Bayesian methods to choose the best model to be used in bioequivalence studies. PMID- 19127467 TI - Test homogeneity of risk difference across subgroups in clinical trials. AB - A weighted least squares statistic is commonly used to test homogeneity of the risk difference for a series of 2 x 2 tables. Since the method is based on asymptotic theory, its type I error rate is inflated when the data are sparse. Two new methods for testing the homogeneity of risk difference across different groups in clinical trials are proposed in this paper. These methods are constructed, based on the Wilson's score test and traditional weighted least squares statistics. The performance of the new methods is evaluated and compared to the currently available approaches. Results show that one of our new methods has a type I error rate that is closest to the nominal level among all the methods and is much more powerful than those proposed by Lipsitz et al. PMID- 19127468 TI - Multiple imputation under the generalized lambda distribution. AB - Although the normality assumption has been regarded as a mathematical convenience for inferential purposes due to its nice distributional properties, there has been a growing interest regarding generalized classes of distributions that span a much broader spectrum in terms of symmetry and peakedness behavior. In this respect, the generalized lambda distribution (GLD) represents a viable choice. In this article, we conduct multiple imputation for univariate continuous data under the GLD to explore the extent to which this procedure works properly; and we make comparisons with normal imputation models via widely accepted accuracy and precision measures using simulated data that exhibit different distributional features as characterized by competing specifications of the third and fourth moments. Furthermore, we present an application using a clinical trials data from psychiatric research. Multiple imputation under the GLD that cover most of the feasible area in the skewness-elongation plane appears to have substantial potential of capturing real missing-data trends that can be encountered in biopharmaceutical practice. PMID- 19127469 TI - The concordance correlation coefficient for repeated measures estimated by variance components. AB - The concordance correlation coefficient (CCC) is an index that is commonly used to assess the degree of agreement between observers on measuring a continuous characteristic. Here, a CCC for longitudinal repeated measurements is developed through the appropriate specification of the intraclass correlation coefficient from a variance components linear mixed model. A case example and the results of a simulation study are provided. PMID- 19127470 TI - Permutation test following covariate-adaptive randomization in randomized controlled trials. AB - In randomized controlled trials, patients are recruited and randomly allocated to treatments. Patients are never randomly sampled from large population of patients on treatments under study. Therefore, it is important to consider the design and statistical analysis based on the randomization model. In this article, we show theoretically that a permutation test based on the difference in means is identical to analysis of covariance if marginal covariate balance is completely attained. Our theoretical results and Monte Carlo simulation study suggest that the permutation test following Pocock-Simon's covariate-adaptive randomization can be a useful alternative to traditional population-based tests in a confirmatory randomized controlled trial with important prognostic factors. The proposed procedure is illustrated with modified data from the randomized placebo controlled trial of pirfenidone. PMID- 19127471 TI - Performance and sample size requirements of Bayesian methods for binary outcomes in fixed-dose combination drug studies. AB - We develop a Bayesian analysis for the study of fixed-dose combinations of two or more drugs. The approach described here does not require knowledge of the dose response relationships of the components or large sample approximations. We provide a procedure to estimate sample size in this context. In addition, we explore the performance of the Bayesian procedure in situations where existing methods are known to perform poorly. PMID- 19127472 TI - Investigating association between behavior, corticosterone, heart rate, and blood pressure in rats using surrogate marker evaluation methodology. AB - The drug development process involves identifying a compound and assessing its merit through rigorous pre-clinical and clinical trials. The pre-clinical stage is designed to assess the chemical properties of the new drug, as well as to determine the steps for synthesis and purification. In this stage of drug development, circumstances might dictate the use of alternative endpoints than the originally anticipated clinically relevant endpoint. In this regard, identification and evaluation of surrogate endpoints is of paramount importance. The validation methods make it possible to quantify degrees of association between the clinically relevant endpoint, also termed the true endpoint, and the alternative, surrogate endpoint. In this paper, we adapt the surrogate marker evaluation methodology of Alonso et al. (2003); (2006), developed for the case of two longitudinal outcomes, to the situation where either a longitudinal surrogate and cross sectional true endpoint is recorded, or vice versa. The work is motivated by a preclinical experiment conducted to assess association between corticosterone (CORT), heart rate, and blood pressure in rats, the data from which are then subjected to analysis. It was found that there is a weak relationship between CORT and behavior, and between CORT on the one hand and heart rate and blood pressure on the other hand, but a reasonably high degree of association was registered between heart rate and behavior. PMID- 19127473 TI - An application of linear mixed effects model to assess the agreement between two methods with replicated observations. AB - We study the problem of assessing the agreement between two methods with any number of replicated observations using linear mixed effects (LME) model with Kronecker product covariance structure in a doubly multivariate set-up. This method can also be used in the case of unbalanced designs when number of replications on each patient is unequal, as well as when the number of replications on each patient by respective methods is unequal. The model is implemented using the MIXED procedure of SAS. We demonstrate our proposed method with three real datasets. PMID- 19127474 TI - An effective approach for obtaining optimal sampling windows for population pharmacokinetic experiments. AB - This paper describes an effective approach for optimizing sampling windows for population pharmacokinetic experiments. Sampling windows has been proposed for population pharmacokinetic experiments that are conducted in late phase drug development programs where patients are enrolled in many centers and out-patient clinic settings. Collection of samples under this uncontrolled environment at fixed times may be problematic and can result in uninformative data. A sampling windows approach is more practicable, as it provides the opportunity to control when samples are collected by allowing some flexibility and yet provide satisfactory parameter estimation. This approach uses D-optimality to specify time intervals around fixed D-optimal time points that results in a specified level of efficiency. The sampling windows have different lengths and achieve two objectives: the joint sampling windows design attains a high specified efficiency level and also reflects the sensitivities of the plasma concentration-time profile to parameters. It is shown that optimal sampling windows obtained using this approach are very efficient for estimating population PK parameters and provide greater flexibility in terms of when samples are collected. PMID- 19127475 TI - Performance of selected nonparametric tests for discrete longitudinal data under different patterns of missing data. AB - Comparison of changes over time of a continuous response variable between treatment groups is often of main interest in clinical trials. When the distributional properties of the continuous response variable are not regular enough, or when the response is discrete, nonparametric techniques have been used. The relative performances of selected repeated measures nonparametric two sample tests proposed by Wei and Lachin, Koziol, Wei and Johnson, and the adapted Wilcoxon Rank-Sum test are compared through simulations based on quality of life data. The Wilcoxon Rank-Sum test is the most powerful and is not significantly affected by the different patterns of missing data. PMID- 19127476 TI - A decision algorithm for translating preclinical trial results to enhance recovery after stroke. AB - A decision algorithm was required to evaluate the first half of a cooperative agreement for preclinical trials to optimize medical device parameters to enhance stroke recovery. Continued funding was contingent upon the midpoint evaluation, called the milestone decision. We developed an algorithm, which summarized our rodent and primate model results. Primary outcomes weighed more heavily than secondary outcomes, and the primate model more heavily than rodent models. By controlling the type I error for this milestone decision, the probability of correctly discontinuing the study if treatment was not beneficial was high (>0.84). Similar algorithms may be adapted for other milestone-driven projects. PMID- 19127480 TI - Equating topical treatment of severe psoriasis to keeping money in a mattress. PMID- 19127481 TI - Synthesis and characterization of some new 2(3H)-benzoxazolones with analgesic and antiinflammatory activities. AB - The synthesis, characterization and pharmacological activities of a new series of (6-difluorobenzoyl)-5-methyl-3-benzoylmethyl-2(3H)-benzoxazolone and 5-methyl-3 (2-hydroxyl-2-phenylethyl)-2(3H)-benzoxazolone are described. Antiinflammatory activity was investigated by the carrageenin-induced paw oedema test and analgesic activity by acetic acid writhing and hot plate tests in mice. Among the synthesized compounds, compound 3e 6-(2,5-difluorobenzoyl)-3-(4 bromobenzoylmethyl-2(3H)-benzoxazolone was found to be the most promising compound for analgesic activity. Reduced compounds (4a-4d) displayed considerable anti-inflammatory activity compared to the other derivatives. PMID- 19127483 TI - Is urological research stalled or stopped? PMID- 19127482 TI - Analysis of chronic lymphotic leukemia transcriptomic profile: differences between molecular subgroups. AB - B cell chronic lymphocytic leukemia (CLL) is a lymphoproliferative disorder with a variable clinical course. Patients with unmutated IgV(H) gene show a shorter progression-free and overall survival than patients with immunoglobulin heavy chain variable regions (IgV(H)) gene mutated. In addition, BCL6 mutations identify a subgroup of patients with high risk of progression. Gene expression was analysed in 36 early-stage patients using high-density microarrays. Around 150 genes differentially expressed were found according to IgV(H) mutations, whereas no difference was found according to BCL6 mutations. Functional profiling methods allowed us to distinguish KEGG and gene ontology terms showing coordinated gene expression changes across subgroups of CLL. We validated a set of differentially expressed genes according to IgV(H) status, scoring them as putative prognostic markers in CLL. Among them, CRY1, LPL, CD82 and DUSP22 are the ones with at least equal or superior performance to ZAP70 which is actually the most used surrogate marker of IgV(H) status. PMID- 19127484 TI - Progress in Alzheimer's disease drug discovery: an update. AB - While Alzheimer's disease (AD) represents a major healthcare challenge, with 25 to 34 million individuals currently affected worldwide and triple this number of patients projected by 2050, the drugs currently approved for the palliative treatment of AD, the cholinesterase inhibitors and the NMDA antagonist memantine, have demonstrated questionable efficacy, highlighting an urgent need for new therapies. Efforts in targeting the removal of amyloid plaques from the brain of patients with AD have been disappointing, with neither plaque-removing vaccines nor the gamma-secretase modulator, tarenflurbil demonstrating clinical benefit, thus questioning the validity of the amyloid cascade hypothesis that has driven AD research for the past decade. The lack of progress in mechanistic approaches (the amyloid and tau hypotheses) to developing new AD drugs indicates that some of the basic assumptions of AD causality and the search for effective drugs are probably in need of major reassessment and redirection. PMID- 19127486 TI - Endothelial dysfunction: A potential therapeutic target for geriatric depression and brain amyloid deposition in Alzheimer's disease? AB - Depression and Alzheimer's disease (AD) are among the most prevalent mental disorders in the elderly. Strong evidence suggests that vascular diseases and vascular risk factors are associated with both depression and AD, and could partially explain the coexistence or the concurrent onset of these two diseases. In particular, endothelial dysfunction appears to play a critical role in the neurobiology of depression and amyloid deposition in the brains of patients with AD. Antidepressants have a significant impact on endothelial function. In addition, several drugs used to treat vascular disease or vascular risk factors, such as calcium-channel blockers, angiotensin-converting enzyme inhibitors and statins, have, to variable extents, significant clinical effects on depressive symptomatology or amyloid deposition in AD. Furthermore, preclinical and clinical data suggest that the nitric oxide and VEGF signaling pathways may be of value for the treatment of depression and AD. PMID- 19127487 TI - The manipulation of cation-chloride co-transporters as a novel means to treat persistent pain, epilepsy and other neurological disorders. AB - Existing members of the anti-epileptic drug (AED) class often fail to provide meaningful symptom relief to patients experiencing persistent pain, epilepsy and other neurological disorders, and can evoke substantial adverse events. In an effort to improve treatment options, much attention has turned to novel mechanisms that may represent points of therapeutic intervention. Among these mechanisms are the cation-chloride co-transporters (CCCs), the dysfunction of which has been linked to aberrant chloride homeostasis in neurons of the CNS, and resulting disorders including persistent pain and epilepsy. This review examines the literature linking CCCs to neurological disease, and discusses their considerable potential as the basis for novel therapeutic strategies. PMID- 19127485 TI - Pharmacotherapy in post-traumatic stress disorder: evidence from randomized controlled trials. AB - This review discusses evidence-based pharmacotherapies for post-traumatic stress disorder (PTSD). The epidemiology of PTSD and its phenomenological characteristics are summarized. Focus is placed on the major classes of drugs for which at least a minimum of evidence-based outcome data are available from randomized controlled trials (RCTs). Drugs for the total symptom constellation of the disorder, specific PTSD symptoms, such as nightmares, and prevention of PTSD development post-trauma, are discussed. Where appropriate, RCT methodological problems that limit the conclusions drawn are discussed. In addition, recommendations for research to fill critical gaps in the knowledge of PTSD treatment are offered. PMID- 19127488 TI - The pharmacological mechanisms of electroacupuncture. AB - Acupuncture is a therapeutic procedure that has been practiced for thousands of years in South East Asia to treat a variety of pathological symptoms and conditions. In 1997, the NIH released a consensus statement concluding that acupuncture is effective or at least useful for the treatment of 13 conditions including nausea and vomiting induced by surgery or chemotherapy, dental pain, and lower back pain. Although the beneficial aspects of acupuncture have been widely accepted, the mechanisms by which acupuncture acts are only partially understood. This review highlights the pharmacological mechanisms underlying the neurological (sensory), cardiovascular and gastrointestinal effects of electroacupuncture. PMID- 19127489 TI - The relationship between dysglycemia and cognitive dysfunction. AB - With the aging of the population, the prevalence of two common disorders is expected to rise: diabetes and dementia. It has been shown that people with diabetes are approximately 1.5-fold more likely to experience cognitive decline and 1.6-fold more likely to develop frank dementia than individuals without diabetes. This is likely due to a higher prevalence of both vascular dementia and Alzheimer's disease. The aim of this review is to describe the importance of this relationship, the evidence supporting it and possible explanations, and the possible role of insulin and insulin resistance in this relationship. PMID- 19127490 TI - Desvenlafaxine, a serotonin-norepinephrine uptake inhibitor for major depressive disorder, neuropathic pain and the vasomotor symptoms associated with menopause. AB - Desvenlafaxine, a serotonin-norepinephrine reuptake inhibitor (SNRI) developed by Wyeth, is a novel salt form of the isolated major active metabolite of the antidepressant venlafaxine. Desvenlafaxine was developed as a slow-release tablet formulation and rapidly penetrates the brain upon administration supporting its direct effects on neuronal systems of the brain. Unlike various other antidepressants including venlafaxine, desvenlafaxine is not metabolized by cytochrome p450 (CYP) enzyme pathways and is associated with minimal inhibition of CYP enzymes. This feature results in a comparatively low risk of drug-drug interaction and consistent intra-individual and inter-individual pharmacokinetic profiles. Desvenlafaxine has been recently approved by the US FDA for the treatment of major depressive disorder (MDD) based on a series of randomized, double-blind, placebo-controlled clinical trials indicating efficacy and safety for patients with MDD. Studies have also supported the potential utility of desvenlafaxine in the treatment of vasomotor symptoms of menopause, anxiety symptoms and painful physical symptoms. However, concerns including mixed efficacy and adverse events need to be further explored in future studies. PMID- 19127491 TI - Gabapentin enacarbil, a gabapentin prodrug for the treatment of the neurological symptoms associated with disorders such as restless legs syndrome. AB - Restless Legs Syndrome (RLS) is a sleep-related movement disorder characterized by an urge to move the legs accompanied by unpleasant symptoms. Dopaminergic agents currently represent the first-line therapy for RLS. However, some patients are unable to continue with this pharmacological approach in the long term because of augmentation or other adverse events. Limited studies with the anticonvulsant/analgesic agent gabapentin have demonstrated that this drug might be useful in the treatment of primary and uremic RLS. Unfortunately, gabapentin has an unfavorable pharmacokinetic profile characterized by a dose-dependent bioavailability. Gabapentin enacarbil, a prodrug of gabapentin, is currently being developed by XenoPort Inc/Astellas Pharma Inc/GlaxoSmithKline plc in order to overcome this limitation. In vitro and in vivo studies have demonstrated that gabapentin enacarbil provides a dose-proportional exposure to gabapentin when orally administered. Gabapentin enacarbil has proved to be beneficial in treating RLS symptoms in several phase II and III clinical trials. In addition, these trials have demonstrated that gabapentin enacarbil is safe and well tolerated, causing transient and mild or moderate adverse events. Gabapentin enacarbil is an interesting compound, which is potentially useful in treating RLS patients who report severe adverse events under dopaminergic agents. The author concludes that additional studies are required in order to better assess the efficacy and safety of gabapentin enacarbil on RLS. PMID- 19127492 TI - Interscience Conference on Antimicrobial Agents and Chemotherapy--48th Annual Meeting and Infectious Diseases Society of America-- 46th Annual Meeting Updates on therapeutic research--Part 1. PMID- 19127493 TI - Interscience Conference on Antimicrobial Agents and Chemotherapy--48th Annual Meeting and Infectious Diseases Society of America-- 46th Annual Meeting Updates on therapeutic research--Part 2. PMID- 19127494 TI - American Heart Association--Scientific Sessions 2008--Studies with investigative drugs--Part 1. PMID- 19127495 TI - American Heart Association--Scientific Sessions 2008--Studies with investigative drugs--Part 2. PMID- 19127496 TI - Society for Neuroscience--38th Annual Meeting--Therapeutics for Parkinson's disease. PMID- 19127497 TI - Society for Neuroscience--38th Annual Meeting--Focus on Huntington's disease. PMID- 19127498 TI - Society for Neuroscience--38th Annual Meeting--Therapeutic approaches for Alzheimer's disease, schizophrenia and bipolar disorder. PMID- 19127499 TI - Society for Neuroscience--38th Annual Meeting--Data for therapeutics for CNS diseases and disorders. PMID- 19127500 TI - Drug Therapy in HIV Infection--Ninth International Congress. PMID- 19127501 TI - Safety Pharmacology World Europe 2008--Healthcare Communications' Second Annual Conference. PMID- 19127502 TI - Targeting transcription factors for cancer therapy. AB - Despite all of the advances in understanding the operations of cancer cells, cancer is on track to pass heart disease as the leading cause of death in the developed world. The primary treatment approach for advanced cancer remains chemotherapeutic drugs that kill cells, both normal and malignant, in a relatively indiscriminate manner. One recent advance in treatment has been the introduction of drugs that target kinases, which can undergo mutations and become activated inappropriately in cancer cells. However, the maximal benefit that can derive from this approach may soon be reached, and a clear need exists for identifying other therapeutic targets that will have broader applicability. Transcription factors, proteins that regulate the expression of genes within a cell and ultimately control cell behavior, are frequently locked in an 'on' position in cancer cells; when the transcription factors are shut off, a cancer cell will generally stop growing or begin to die. In contrast, normal cells tolerate a loss of transcription factor function with little consequence, as a result of redundancies that are present under normal conditions. Despite challenges involved in developing drugs to target transcription factors, increasing evidence indicates that this approach may be a highly promising strategy in the development of cancer treatments that are more effective and less toxic than existing options. PMID- 19127503 TI - Predictive biomarkers for EGFR therapy. AB - The results of the National Cancer Institute of Canada Clinical Trials Group (NCIC CTG) BR.21 study, a phase III, randomized, placebo-controlled trial of erlotinib in patients with advanced NSCLC who had failed first-line or second line chemotherapy, provided new treatment options for lung cancer. Given that only a small percentage of patients may benefit from treatment with inhibitors that target the EGF receptor (EGFR), substantial effort has been devoted to identifying biomarkers that are predictive of the highest likelihood of benefit from these drugs. Several markers, including EGFR tyrosine kinase domain mutation, EGFR gene copy number and KRAS mutation, have been investigated extensively, but results obtained to date remain controversial. This feature review discusses available data and the basis for the controversies. PMID- 19127504 TI - Laromustine, a sulfonyl hydrolyzing alkylating prodrug for cancer therapy. AB - Laromustine (Onrigin), under development by Vion Pharmaceuticals Inc, belongs to the sulfonylhydrazine class of alkylating agents and is in clinical development for the treatment of malignancies. Laromustine is a prodrug that yields a chloroethylating compound (VNP-4090-CE) and a carbamoylating compound (methyl isocyanate). The antineoplastic effect of laromustine is attributed primarily to the chloroethylating species, which causes the preferential alkylation of DNA at the O6 position of guanine, a lesion that results in interstrand crosslinks and, eventually, cell death. The carbamoylating species contributes to antitumor activity by inhibiting the DNA repair protein O6-alkylguanine transferase. Early phase I clinical trials in patients with solid tumors indicated that laromustine was associated with myelosuppression; few extramedullary toxicities were observed, indicating potential efficacy for the treatment of hematological malignancies. Phase II trials have been completed in patients with previously untreated acute myelogenous leukemia (AML), high-risk myelodysplastic syndrome (MDS) and relapsed AML. The most encouraging results were observed in patients over 60 years of age with poor-risk de novo AML for which no standard treatment exists. Laromustine is currently in phase II/III trials for AML and phase II trials for MDS and solid tumors. Laromustine appears to be a promising agent that will add to the armamentarium of drugs available to treat patients who do not respond to, or are not fit for, intensive chemotherapy, such as elderly individuals. PMID- 19127505 TI - [Quasi mortis imago]. PMID- 19127506 TI - [Long-term service-dependent patients in mental health care: connection to urbanisation]. AB - BACKGROUND: In view of the increased rate of psychiatric morbidity in urban areas there is likely to be a relation between urbanisation and the extent to which patients use the care services. It is not yet known whether this relation is particularly strong in the case of long-term service-dependent patients with mental health problems. AIM: To investigate the possible relation between the prevalence of long-term service-dependent patients and the degree of urbanisation and to find out if there are differences in the strength of this relation in the case of different types of long-term patients and between these patients and other psychiatric patients. METHOD: Data were collected from three psychiatric case registers and from the Statistics Netherlands (cbs). Distinctions were made between three types of long-term patients: 'long stay', 'residential & outpatient' and 'outpatient'. The population was divided into five categories according to the degree of urbanisation in the area of residence. Data were analysed using Poisson regression analysis, corrected for differences in age, sex, marital status and the amount of care available. RESULTS: The incidence rate ratios for the categories 'long stay', 'residential & outpatient' and 'outpatient' and for other patients were significantly higher for each category of urbanisation than for non-urbanised municipalities. There was also a significant positive linear relation. For long stay patients no significant relation was found with the degree of urbanisation. The total ratios for long term patients were higher than the ratios for other types of patients. The ratios for residential and outpatient were higher than the ratios for outpatient. CONCLUSION: The prevalence of long-term service-dependent patients in mental health care increases with the degree of urbanisation and this increase is stronger than the increase observed for other patients. Furthermore, the relation with urbanisation for the category residential and outpatient is stronger for the category residential and outpatient than for the long-term outpatients. PMID- 19127507 TI - [Gene-environment interaction in psychiatry: new views of the influence of early relationships]. AB - The results of studies on the interaction of genes with the environment seem to take the 'nature nurture' discussion to higher ground. Genes and the environment influence each other via complex processes. Psychopathology apparently develops as a result of a person's genetic make-up which can influence behaviour only in the context of specific environmental factors. Or, you can turn the argument around: environmental factors only lead to psychopathology if someone has a specific genetic make-up. Genes and the environment are therefore dependent on each other and interact on a daily basis. People, just like plants and animals, are finely tuned so that adaptation can occur. The quality of early relationships plays a decisive role in this tuning. In the light of recent progress in the neurosciences, particularly in the field of biological models of early development, there is a need for conceptual adaptation in psychiatry. PMID- 19127508 TI - [Nothing is carved in stone]. PMID- 19127509 TI - [Reaction to 'Severe mental illness: cure and care?']. PMID- 19127510 TI - [Reaction to 'The judge follows the advice of the psychiatrist; an investigation into the jurisprudence concerning compulsory admission requests and decisions under the Dutch Act on Special Admissions to Psychiatric Hospitals (Bopz)]. PMID- 19127511 TI - [Reaction to 'Thiamine deficiency caused by malnutrition: a rare cause?']. PMID- 19127512 TI - Profiling microRNA expression: a snapshot of nonalcoholic steatohepatitis and a recording of its pathogenesis. PMID- 19127513 TI - Sustained virological response reduces incidence of onset of type 2 diabetes in chronic hepatitis C. AB - Diabetes is present in patients with chronic hepatitis C virus infection. The aim of this retrospective cohort study was to assess the cumulative development incidence and predictive factors for type 2 diabetes after the termination of interferon therapy in Japanese patients positive for hepatitis C virus (HCV). A total of 2,842 HCV-positive patients treated with interferon (IFN) monotherapy or combination therapy with IFN and ribavirin were enrolled. The mean observation period was 6.4 years. An overnight (12-hour) fasting blood sample or a casual blood sample was taken for routine analyses during follow-up. The primary goal was the onset of type 2 diabetes. Evaluation was performed by using the Kaplan Meier method and Cox proportional hazard analysis. Of 2,842 HCV patients, 143 patients developed type 2 diabetes. The cumulative development rate of type 2 diabetes was 3.6% at 5 years, 8.0% at 10 years, and 17.0% at 15 years. Multivariate Cox proportional hazard analysis revealed that type 2 diabetes development after the termination of IFN therapy occurred when histological staging was advanced (hazard ratio 3.30; 95% confidence interval [CI] 2.06-5.28; P < 0.001), sustained virological response was not achieved (hazard ratio 2.73; 95% CI 1.77-4.20; P < 0.001), the patient had pre-diabetes (hazard ratio 2.19; 95% CI 1.43-3.37; P < 0.001), and age was >or=50 years (hazard ratio 2.10; 95% CI 1.38-3.18; P < 0.001). CONCLUSION: Our results indicate sustained virological response causes a two-thirds reduction in the risk of type 2 diabetes development in HCV-positive patients treated with IFN. PMID- 19127514 TI - Ag-doped manganite nanoparticles: new materials for temperature-controlled medical hyperthermia. AB - The purpose of this study was to introduce newly synthesized nanomaterials as an alternative to superparamagnetic ironoxide based particles (SPIO) and thus to launch a new platform for highly controllable hyperthermia cancer therapy and imaging. The new material that forms the basis for this article is lanthanum manganite particles with silver ions inserted into the perovskite lattice: La(1 x)Ag(x)MnO(3+delta). Adjusting the silver doping level, it is possible to control the Curie temperature (T(c)) in the hyperthermia range of interest (41-44 degrees C). A new class of nanoparticles based on silver-doped manganites La(1 x)Ag(x)MnO(3+delta) is suggested. New nanoparticles are stable, and their properties were not affected by the typical ambient conditions in the living tissue. It is possible to monitor the particle uptake and retention by MRI. When these particles are placed into an alternating magnetic field, their temperature increases to the definite value near T(c) and then remains constant if the magnetic field is maintained. During the hyperthermia procedure, the temperature can be restricted, thereby preventing the necrosis of normal tissue. A new class of nanoparticles based on silver-doped manganites La(1-x)Ag(x)MnO(3+delta) was suggested. Ag-doped perovskite manganites particles clearly demonstrated the effect of adjustable Curie temperature necessary for highly controllable cellular hyperthermia. The magnetic relaxation properties of the particles are comparable with that of SPIO, and so we were able to monitor the particle movement and retention by MRI. Thus, the new material combines the MRI contrast enhancement capability with targeted hyperthermia treatment. PMID- 19127515 TI - Keyhole cochlear implantation surgery. AB - A limited access 'keyhole' cochlear implantation (CI) surgical technique is presented. The surgery employs a 15-18 mm diameter C-incision on the rear of the auricle. The implant is retained in a snug pericranial pocket. A bony retention well is avoided. Minimal mastoid cell clearance is used. The surgery is brief, atraumatic and well suited to small infant cases. Used in 315 sequential routine CI cases from 1997-2007, the complication rates compare favourably with past reports. The outcomes of the series question the use of larger incisions and implant retention wells. Current implant designs should be modified for better adaptation to the cranial contours. PMID- 19127516 TI - Hepatic steatosis in response to acute alcohol exposure in zebrafish requires sterol regulatory element binding protein activation. AB - Steatosis is the most common consequence of acute alcohol abuse and may predispose to more severe hepatic disease. Increased lipogenesis driven by the sterol response element binding protein (SREBP) transcription factors is essential for steatosis associated with chronic alcohol ingestion, but the mechanisms underlying steatosis following acute alcohol exposure are unknown. Zebrafish larvae represent an attractive vertebrate model for studying alcoholic liver disease (ALD), because they possess the pathways to metabolize alcohol, the liver is mature by 4 days post-fertilization (dpf), and alcohol can be simply added to their water. Exposing 4 dpf zebrafish larvae to 2% ethanol (EtOH) for 32 hours achieves approximately 80 mM intracellular EtOH and up-regulation of hepatic cyp2e1, sod, and bip, indicating that EtOH is metabolized and provokes oxidant stress. EtOH-treated larvae develop hepatomegaly and steatosis accompanied by changes in the expression of genes required for hepatic lipid metabolism. Based on the importance of SREBPs in chronic ALD, we explored the role of Srebps in this model of acute ALD. Srebp activation was prevented in gonzo larvae, which harbor a mutation in the membrane-bound transcription factor protease 1 (mbtps1) gene, and in embryos injected with a morpholino to knock down Srebp cleavage activating protein (scap). Both gonzo mutants and scap morphants were resistant to steatosis in response to 2% EtOH, and the expression of many Srebp target genes are down-regulated in gonzo mutant livers. CONCLUSION: Zebrafish larvae develop signs of acute ALD, including steatosis. Srebp activation is required for steatosis in this model. The tractability of zebrafish genetics provides a valuable tool for dissecting the molecular pathogenesis of acute ALD. PMID- 19127518 TI - East-West pathology agreement on precancerous liver lesions and early hepatocellular carcinoma. PMID- 19127517 TI - Knockout of myeloid cell leukemia-1 induces liver damage and increases apoptosis susceptibility of murine hepatocytes. AB - Myeloid cell leukemia-1 (Mcl-1) is an antiapoptotic member of the Bcl-2 protein family. It interacts with proapoptotic Bcl-2 family members, thereby inhibiting mitochondrial activation and induction of apoptosis. Mcl-1 is essential for embryonal development and the maintenance of B cells, T cells, and hematopoietic stem cells. We have recently shown that induction of Mcl-1 by growth factors rescues primary human hepatocytes from CD95-mediated apoptosis. This prompted us to further analyze the relevance of Mcl-1 for hepatocellular homeostasis. Therefore, we generated a hepatocyte-specific Mcl-1 knockout mouse (Mcl 1(flox/flox)-AlbCre). Deletion of Mcl-1 in hepatocytes results in liver cell damage caused by spontaneous induction of apoptosis. Livers of Mcl-1(flox/flox) AlbCre mice are smaller compared to control littermates, due to higher apoptosis rates. As a compensatory mechanism, proliferation of hepatocytes is enhanced in the absence of Mcl-1. Importantly, hepatic pericellular fibrosis occurs in Mcl-1 negative livers in response to chronic liver damage. Furthermore, Mcl 1(flox/flox)-AlbCre mice are more susceptible to hepatocellular damage induced by agonistic anti-CD95 antibodies or concanavalin A. CONCLUSION: The present study provides in vivo evidence that Mcl-1 is a crucial antiapoptotic factor for the liver, contributing to hepatocellular homeostasis and protecting hepatocytes from apoptosis induction. PMID- 19127519 TI - Hepatic function is preserved in the absence of mature microRNAs. AB - MicroRNAs (miRNAs) are small noncoding RNA molecules that regulate gene expression through partial or complete complementarity with target messenger RNAs. The function of miRNAs in normal liver physiology is largely unknown. We address the role of Dicer1 in the differentiated liver. We derived mice lacking Dicer1 function in hepatocytes and assessed the loss of mature miRNA via quantitative polymerase chain reaction. Gene expression microarray analysis was performed on liver RNA from mutant and control mice. Liver sections from mutant and control mice were examined and liver function tests were performed. Mice lacking Dicer1 function in hepatocytes appeared and behaved normally. Despite the loss of mature miRNAs, hepatic function was maintained, as reflected by normal blood glucose, albumin, cholesterol, and bilirubin. However, mutant mice between 2 and 4 months of age exhibited progressive hepatocyte damage with elevated serum alanine aminotransferase and aspartate aminotransferase. Liver mass was increased in mutant mice, as were cellular markers of both proliferation and apoptosis. Microarray analysis indicated large-scale changes in gene expression, with increased expression of many miRNA targets, particularly imprinted genes. CONCLUSIONS: Loss of miRNA processing in the liver at late gestation has a remarkably mild phenotype, suggesting that miRNAs do not play an essential role in hepatic function. However, miRNA deficiency results in hepatocyte apoptosis, hepatocyte regeneration, and portal inflammation. Finally, microarray analysis of gene expression in the mutant liver supports a previously hypothesized role for Dicer1 in the repression of imprinted genes. PMID- 19127521 TI - Parkinson's disease and suicide: a profile of suicide victims with Parkinson's disease in a population-based study during the years 1988-2002 in Northern Finland. AB - OBJECTIVE: We studied the prevalence of hospital-treated Parkinson's disease (PD) among suicide victims and the profile of these persons, taking into account suicide attempts, timing of depression and comorbid somatic diseases. METHODS: The database of this study consisted of suicide victims aged 50 years of age or older (n = 555) during a fourteen-year period in the province of Oulu in Northern Finland. RESULTS: Hospital-treated Parkinson's disease occurred in 1.6% of the subjects, indicating a rather low prevalence of suicide in this group of patients. The persons with PD had attempted suicide earlier in 44% of the cases, while the corresponding percentage for other victims in older age was 9.9% (p = 0.009 Fischer exact test). CONCLUSIONS: Based on the case characteristics of our study the profile of PD person who completed suicide was as follows: male subject with recently diagnosed disease, living in rural area, having multiple physical illnesses, and having attempted suicide earlier. Psychiatric consultation is thus highly recommended for the PD patients with this disease profile. PMID- 19127520 TI - Gender differences in trochlear groove orientation and rotational kinematics of human knees. AB - Knowledge of the morphology and kinematics of the male and female knees is important for understanding gender-related dimorphism in knee pathology and improvement of related surgical treatments. Twelve male and 12 female subjects with healthy knees were recruited, and each subject performed a single leg lunge while images of the knee were recorded by two fluoroscopes. Tibiofemoral joint motion was then reproduced using bony models matched to the fluoroscopic images. Femoral trochlear groove orientation was also measured in each knee. While many of the measured parameters were found to be similar between the genders, a few interesting differences were also noted. Females showed greater external tibial rotation at 0 degrees flexion (-5.4 degrees vs. -1.3 degrees, p = 0.03), smaller internal rotation at 30 degrees flexion (1.7 degrees vs. 6.4 degrees, p = 0.04) and greater range of tibial rotation (18.2 degrees vs. 12.4 degrees, p = 0.01) compared to males. Female knees also had a more medially oriented proximal trochlear groove (10.0 degrees vs. 4.5 degrees, p = 0.04). These gender differences in rotational kinematics and trochlear groove orientation may warrant further studies to determine implications for surgical treatments such as total knee arthroplasty, and gender-related dimorphism in certain knee injuries and pathologies, like anterior cruciate ligament injury and patellofemoral problems. PMID- 19127522 TI - Regressive events in rat corticospinal axons during development in in vitro slice cocultures: retraction, amputation, and degeneration. AB - Axonal regression is utilized to refine neuronal circuits during development, but the dynamic properties of such regression remain largely unknown. We used confocal time-lapse imaging to examine the regression of single enhanced yellow fluorescent protein-labeled axons in corticospinal slice cocultures. By acquiring images at long (1 day) and short (30-60 min) intervals on days 5-13 in vitro, we detected three types of regressive events: 1) retractions, 2) amputations (referred to as autoaxotomy), and 3) degeneration. Retractions proceeded at some constant rate for up to 3 hours and then paused or switched to another constant rate, apparently shifting stepwise among three retraction rates (6, 12, 17 microm/hours). Autoaxotomy was a previously unreported strategy for regression. It occurred spontaneously, either at a proximal branch neck or at a distal end. Axons also underwent a form of degeneration that had several novel characteristics. Degenerating axons showed bright bead-like spots arranged at 3-9 microm intervals. The gaps were much larger than the spot size, and there was no prior sign of damage (e.g., swelling). Each spot's fluorescence intensity often waxed and waned, with its position unchanged. Degeneration progressed without clear proximal-to-distal directionality and was complete within 3-4 hours. Quantitative analysis of daily branch regression showed that branches almost always regressed up to their branch point or stopped before it, thereby keeping the branch point stable. This branch-point stability was retained for all three regression strategies observed, suggesting that the fate of each branch is determined relatively independently during the development of axonal arborization. PMID- 19127523 TI - Involvement of R-type Ca2+ channels in neurotransmitter release from spinal dorsolateral funiculus terminals synapsing motoneurons. AB - Molecular studies have revealed the presence of R-type voltage-gated Ca(2+) channels at pre- and postsynaptic regions; however, no evidence for the participation of these channels in transmitter release has been presented for the spinal cord. Here we characterize the effects of SNX-482, a selective R channel blocker, on the monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in motoneurons by stimulation of dorsolateral funiculus (DLF) terminals in a slice preparation from the adult turtle spinal cord. SNX-482 inhibited neurotransmission in a dose-dependent manner, with an IC(50) of approximately 9 +/- 1 nM. The EPSP time course and membrane time constant of the motoneurons were not altered, suggesting a presynaptic mechanism. The toxin inhibited the residual component of the EPSPs recorded in the presence of N- and P/Q-type Ca(2+) channel blockers, strongly suggesting a role for the R channels in neurotransmission at the spinal cord DLF terminals. Consistently with this, RT-PCR analysis of turtle spinal cord segments revealed the expression of the Ca(V)2.3 pore-forming (alpha(1E)) subunit of R channels, whereas the use of anti-alpha(1E)-specific antibodies resulted in its localization in the DLF fibers as demonstrated by immunohistochemistry coupled with laser confocal microscopy. PMID- 19127524 TI - The use of benzodiazpines and related drugs amongst older people in Sweden: associated factors and concomitant use of other psychotropics. AB - OBJECTIVE: To determine the factors associated with use of benzodiazepines and benzodiazepine related drugs (BZDs/BZDRDs) in a nationwide population of older people. METHODS: We analyzed data on age, sex, type of residential area (urban/rural), and dispensed drugs for people aged > or = 75 years registered in the Swedish Prescribed Drug Register from October to December 2005 (n = 731,105; corresponds to 91% of the domain population). Multivariate logistic regression analysis was used for analyzing whether age, sex, type of residential area, number of other drugs, and concomitant psychotropic drug use was associated with use of BZDs/BZDRDs. RESULTS: > or =1 BZDs/BZDRDs and > or =2 BZDs/BZDRDs were used by 25% and 5% of the study population, respectively. The most commonly used BZDs/BZDRDs were zopiclone, zolpidem, oxazepam, flunitrazepam, and diazepam. The probability of use BZDs/BZDRDs increased with age, female gender, living in an urban area, use of many other drugs, and concomitant use of other psychotropics, particularly antidepressants. In the comparisons of different BZDs/BZDRDs (n = 179,632), the medium-acting BZDs were associated with higher age, female gender, and use of other psychotropics, whereas BZDRDs showed the opposite pattern. Also, the two BZDRDs zopiclone and zolpidem showed different patterns regarding age, sex, and use of other drugs. CONCLUSIONS: One in four of the elderly used one or more BZD/BZDRD, which should be prescribed cautiously and with the recognition of the potentially negative effects. There were differences in patterns of use for different BZDs/BZDRDs, even within the same drug class. Hence, large study populations are needed for future research of individual BZDs/BZDRDs. PMID- 19127526 TI - No place like home: testosterone responses to victory depend on game location. AB - Several studies have demonstrated that a variety of factors influence testosterone responses to competitive interactions. This study examined the extent to which game location would influence testosterone responses to human competition. Male amateur ice hockey players (n = 10) provided saliva samples before and after competing against the same opponent on two separate occasions (one game at home and one game away). Although both games resulted in similar victories, the home victory was associated with a significantly larger rise in testosterone concentrations relative to the away victory. The factors responsible for the different testosterone responses are not known, however, it is possible that a rise in status in front of the home crowd is more rewarding to athletes, and thus, a more potent stimulus for the endocrine system. PMID- 19127525 TI - Year-round high physical activity levels in agropastoralists of Bolivian Andes: results from repeated measurements of DLW method in peak and slack seasons of agricultural activities. AB - By the repeated use of the doubly labeled water method (DLW), this study aimed to investigate (1) the extent of changes in energy expenditure and physical activity level (PAL) in response to increased agricultural work demands, and (2) whether the seasonal work demands induce the changes in the fairly equitable division of work and similarity of energy needs between men and women observed in our previous study (Phase 1 study; Kashiwazaki et al., 1995: Am J Clin Nutr 62: 901 910). In a rural small agropastoral community of the Bolivian Andes, we made the follow-up study (Phase 2, 14 adults; a time of high agricultural activity) of the Phase 1 study (12 adults; a time of low agricultural activity). In the Phase 2 study, both men and women showed very high PAL (mean+/-SD), but there was no significant difference by sex (men; 2.18 +/- 0.23 (age; 64 +/- 11 years, n = 7), women; 2.26 +/- 0.25 (63 +/- 10 years, n = 7)). The increase of PAL by 11% (P = 0.023) in the Phase 2 was equally occurred in both men and women. The factorial approach underestimated PAL significantly by approximately 15% (P < 0.05). High PAL throughout the year ranging on average 2.0 and 2.2 was attributable to everyday tasks for subsistence and domestic works undertaking over 9-11 h (men spent 2.7 h on agricultural work and 4.7 h on animal herding, whereas women spent 7.3 h almost exclusively on animal herding). The seasonal increase in PAL was statistically significant, but it was smaller than those anticipated from published reports. A flexible division of labor played an important role in the equitable energetic increase in both men and women. PMID- 19127527 TI - Is puberty starting earlier in urban South Africa? AB - Age at the initation of pubertal development was estimated for 401 Black (212 boys) and 206 White (100 boys) urban South African adolescents born in Soweto Johannesburg in 1990. Average age at the initation of puberty, assessed by age at the transition from Tanner Stage 1 to Tanner Stage 2 for breast/genitalia or pubic hair development ranged between 9.8 and 10.5 years. There were no statistically significant differences in age at initiation between genders or ethnic groups. Age at the initation of pubertal development has remained stable over the last 10 to 15 years, with the exception of pubic hair in boys which has declined on average 1.3 years over a decade. There is evidence to suggest that the tempo of pubertal maturation is increasing in girls born in the Soweto Johannesburg area, however, the evidence is less clear for boys. PMID- 19127528 TI - Conformational analysis and stereochemical dependences of (31)P-(1)H spin-spin coupling constants of bis(2-phenethyl)vinylphosphine and related phosphine chalcogenides. AB - Theoretical energy-based conformational analysis of bis(2 phenethyl)vinylphosphine and related phosphine oxide, sulfide and selenide synthesized from available secondary phosphine chalcogenides and vinyl sulfoxides is performed at the MP2/6-311G** level to study stereochemical behavior of their (31)P-(1)H spin-spin coupling constants measured experimentally and calculated at different levels of theory. All four title compounds are shown to exist in the equilibrium mixture of two conformers: major planar s-cis and minor orthogonal ones, while (31)P-(1) H spin-spin coupling constants under study are found to demonstrate marked stereochemical dependences with respect to the geometry of the coupling pathways, and to the internal rotation of the vinyl group around the P(X)-C bonds (X = LP, O, S and Se), opening a new guide in the conformational studies of unsaturated phosphines and phosphine chalcogenides. PMID- 19127530 TI - Fabric softeners: nearly instantaneous characterization and quality control of cationic surfactants by easy ambient sonic-spray ionization mass spectrometry. AB - A tiny droplet of typical samples of fabric softeners from different commercial brands placed on a smooth paper surface was subjected to easy ambient sonic-spray ionization mass spectrometry (EASI-MS). With no need for sample-preparation or pre-separation procedures, EASI-MS and EASI-MS/MS identify nearly instantaneously the main surfactants and the homologous series employed in their formulations. Adulterated and low quality samples containing no or less efficient softeners are also easily recognized. PMID- 19127529 TI - The impact of patient-perceived restricted access to anti-TNF therapy for rheumatoid arthritis: a qualitative study. AB - OBJECTIVE: To explore rheumatoid arthritis (RA) patients' experience of access to anti-tumour necrosis factor (anti-TNF) therapy in the UK, and of switching therapies after an initial failure. METHODS: Patients were asked about their experience of accessing, receiving and discontinuing anti-TNF therapy in face-to face indepth interviews, within the context of the larger study about treatment outcomes. Seventeen individuals with a diagnosis of RA and experience of receiving anti-TNF therapy were interviewed in one hospital trust in England. RESULTS: Different emotions (Theme 1) surrounded the process of accessing anti TNF therapy: hope, desperation, apprehension, anxiety and frustration. Experience of receiving anti-TNF therapy (Theme 2) included not only positive transformation, but also fear of failure and discontinuation. The subsequent value that patients placed on anti-TNF therapy (Theme 3) included having a right to receive therapy and being lucky. These three themes were underpinned by the core category of 'willing to try anything'. Those switching therapies reported increased caution over the possibility of recurring side effects, but some suggestion of benefit. There was a perception that access to anti-TNF therapy was restricted by cost, rather than being recommended for those in clinical need. CONCLUSIONS: Anti-TNF therapies may have a sudden and dramatic impact on RA patients' lives that contrast with other available medications. However, the stress of the patient's journey through the need to 'qualify' for anti-TNF therapy, and the fear of failing or discontinuation of therapy, should not be underestimated by clinicians. PMID- 19127531 TI - Speciation of Zn-aminopolycarboxylic complexes by electrospray ionization mass spectrometry and ion chromatography with inductively coupled plasma mass spectrometry. AB - The speciation of Zn-aminopolycarboxylic complexes was investigated using both electrospray ionization mass spectrometry (ESI-MS) and ion chromatography (IC) with inductively coupled plasma mass spectrometry (ICP-MS). The resulting ESI mass spectra indicated that [Zn(HEDTA)](1-), [Zn(NTA)](1-), [Zn(EDTA)](2-) and [Zn(DTPA)](3-) were all simultaneously detected in solution; [Zn(NTA)](1-) exhibited the weakest intensity of all these Zn-aminopolycarboxylic complexes. IC/ICP-MS was also successfully used to separate Zn complexes by anion-exchange chromatography using a mobile phase containing 30 mM (NH(4))(2)HPO(4) at pH 7.5 giving reasonable resolution within 15 min. A weak peak attributable to the poor stability [Zn(NTA)](1-) ion was also observed using IC/ICP-MS. With the exception of [Zn(NTA)](1-), detection limits ranging from 0.5 to 1.0 microg/L were obtained and the proposed method was used for the determination of Zn aminopolycarboxylic complexes in soil solution. PMID- 19127532 TI - Sensory loss in multifocal motor neuropathy: a clinical and electrophysiological study. AB - Some patients fulfilling the criteria for the diagnosis of multifocal motor neuropathy with conduction block (MMN-CB) at the onset of disease may subsequently develop a sensory loss associated with electrophysiological sensory abnormalities. The latter could represent an overlap between MMN-CB and multifocal acquired demyelinating sensory and motor (MADSAM) neuropathy. The objective was to specify the features of MMN-CB with sensory loss (MMN-CB-Se). Five patients in a series of 11 consecutive patients who fulfilled the criteria of the American Association of Neuromuscular and Electrodiagnostic Medicine for MMN-CB at the first examination and were treated periodically with intravenous immunoglobulin (IVIg) developed sensory loss in the course of the disease. In these five patients we compared the clinical, laboratory, and electrophysiological features found after the development of sensory loss with those at the first examination. The mean time to appearance of objective sensory signs was 7.2 years. In three of the five patients the sensory loss was preceded by intermittent paresthesias in the same nerve territories as the motor involvement. The most frequent electrophysiological abnormality was amplitude reduction of sensory nerve action potentials. There were no bilateral or symmetrical clinical and electrophysiological sensory abnormalities. Anti-GM1 IgM antibodies were positive in four patients. MMN-CB-Se could be an overlap between MMN-CB and MADSAM. It shares the distribution of the sensory disorders encountered in MADSAM, but it is closer to MMN-CB on clinical and therapeutic levels. Study of more patients would be useful to classify this subgroup more accurately. PMID- 19127533 TI - Accessory nerve lesion after cervicofacial lift: clinical and electrodiagnostic evaluations of two cases. AB - Two cases of accessory nerve lesion are reported that occurred within 2.5 weeks and 4.5 weeks, respectively, after surgery for cervicofacial lift. The patients were referred for electrodiagnostic examination because of persistent and unexplained unilateral shoulder pain and disability, 6 and 5 months, respectively, after face lift. In both cases clinical examination revealed severe right-shoulder weakness related to trapezius palsy, without trapezius muscle atrophy in the first case and with trapezius atrophy in the second. Electrodiagnosis revealed bilateral accessory nerve lesions in the first case and a unilateral lesion in the second case. Recovery for both cases was progressive but delayed, and both were evaluated 11 and 10 months, respectively, after the first evaluation. Clinical and especially electrodiagnostic findings suggested that the accessory nerve lesion was related to conduction block in the first case and severe axonal loss in the second case. PMID- 19127534 TI - Racial differences in myasthenia gravis in Alabama. AB - Demographic, clinical, and laboratory features were compared in 235 white and African-American (AA) patients with myasthenia gravis (MG) at the University of Alabama at Birmingham Neuromuscular Disease Clinic from May 2003 to January 2008. Seventy nine percent of patients were white. Acetylcholine receptor antibody was positive in 71% of white patients and in 59% of AA. In patients with seronegative generalized MG, the rate of positive muscle-specific tyrosine kinase antibody (MuSK-Ab) was significantly higher in AA than it was in whites (50% in AA vs. 17% in whites). Ocular MG was seronegative in 75% of AA patients. In AA, MG occurred earlier and more frequently in females, whereas, in whites, disease onset was later and more common in males. Another significant difference was a higher percentage of abnormality on repetitive nerve stimulation in AA. There was also a tendency for more severe forms of MG in AA. There are racial differences in MG between whites and AA in Alabama. These racial differences highlight the need to study biological factors in the pathogenesis of MG and to assess different approaches in diagnosis and treatment. PMID- 19127535 TI - ACCF/SCAI/STS/AATS/AHA/ASNC 2009 Appropriateness Criteria for Coronary Revascularization : a report of the American College of Cardiology Foundation Appropriateness Criteria Task Force, Society for Cardiovascular Angiography and Interventions, Society of Thoracic Surgeons, American Association for Thoracic Surgery, American Heart Association, and the American Society of Nuclear Cardiology. Endorsed by the American Society of Echocardiography, the Heart Failure Society of America, and the Society of Cardiovascular Computed Tomography. AB - The American College of Cardiology Foundation (ACCF), Society for Cardiovascular Angiography and Interventions, Society of Thoracic Surgeons, and the American Association for Thoracic Surgery, along with key specialty and subspecialty societies, conducted an appropriateness review of common clinical scenarios in which coronary revascularization is frequently considered. The clinical scenarios were developed to mimic common situations encountered in everyday practice and included information on symptom status, extent of medical therapy, risk level as assessed by noninvasive testing, and coronary anatomy. Approximately 180 clinical scenarios were developed by a writing committee and scored by a separate technical panel on a scale of 1 to 9. Scores of 7 to 9 indicate that revascularization was considered appropriate and likely to improve health outcomes or survival. Scores of 1 to 3 indicate revascularization was considered inappropriate and unlikely to improve health outcomes or survival. The mid range (4 to 6) indicates a clinical scenario for which the likelihood that coronary revascularization would improve health outcomes or survival was considered uncertain. For the majority of the clinical scenarios, the panel only considered the appropriateness of revascularization irrespective of whether this was accomplished by percutaneous coronary intervention (PCI) or coronary artery bypass graft surgery (CABG). In a select subgroup of clinical scenarios in which revascularization is generally considered appropriate, the appropriateness of PCI and CABG individually as the primary mode of revascularization was considered. In general, the use of coronary revascularization for patients with acute coronary syndromes and combinations of significant symptoms and/or ischemia was viewed favorably. In contrast, revascularization of asymptomatic patients or patients with low-risk findings on noninvasive testing and minimal medical therapy were viewed less favorably. It is anticipated that these results will have an impact on physician decision making and patient education regarding expected benefits from revascularization and will help guide future research. PMID- 19127537 TI - Survivor research delves into late effects of treatment: follow-up treatment plans are a key component. PMID- 19127536 TI - Revitalization of a diastemal tooth primordium in Spry2 null mice results from increased proliferation and decreased apoptosis. AB - An understanding of the factors that promote or inhibit tooth development is essential for designing biological tooth replacements. The embryonic mouse dentition provides an ideal system for studying such factors because it consists of two types of tooth primordia. One type of primordium will go on to form a functional tooth, whereas the other initiates development but arrests at or before the bud stage. This developmental arrest contributes to the formation of the toothless mouse diastema. It is accompanied by the apoptosis of the rudimentary diastemal buds, which presumably results from the insufficient activity of anti-apoptotic signals such as fibroblast growth factors (FGFs). We have previously shown that the arrest of a rudimentary tooth bud can be rescued by inactivating Spry2, an antagonist of FGF signaling. Here, we studied the role of the epithelial cell death and proliferation in this process by comparing the development of a rudimentary diastemal tooth bud (R(2)) and the first molar in the mandibles of Spry2(-/-) and wild-type (WT) embryos using histological sections, image analysis and 3D reconstructions. In the WT R(2) at embryonic day 13.5, significantly increased apoptosis and decreased proliferation were found compared with the first molar. In contrast, increased levels of FGF signaling in Spry2(-/-) embryos led to significantly decreased apoptosis and increased proliferation in the R(2) bud. Consequently, the R(2) was involved in the formation of a supernumerary tooth primordium. Studies of the revitalization of rudimentary tooth primordia in mutant mice can help to lay the foundation for tooth regeneration by enhancing our knowledge of mechanisms that regulate tooth formation. PMID- 19127538 TI - Long-term incidence of cervical cancer in women with human immunodeficiency virus. AB - BACKGROUND: The objective of this study was to estimate the incidence of invasive cervical cancer (ICC) in women with human immunodeficiency virus (HIV) and compare it with the incidence in HIV-uninfected women. METHODS: In a cohort study of HIV-infected and uninfected women who had Papanicolaou tests obtained every 6 months, pathology reports were retrieved for women who had biopsy results or a self-report of ICC. Histology was reviewed when reports confirmed ICC. Incidence rates were calculated and compared with those in HIV-negative women. RESULTS: After a median follow-up of 10.3 years, 3 ICCs were confirmed in HIV-seropositive women, and none were confirmed in HIV-seronegative women. The ICC incidence rate was not found to be associated significantly with HIV status (HIV-negative women [0 of 100,000 person-years] vs HIV-positive women [21.4 of 100,000 person-years]; P = .59). A calculated incidence rate ratio standardized to expected results from the Surveillance Epidemiology and End Results database that was restricted to HIV infected Women's Interagency HIV Study participants was 1.32 (95% confidence interval, 0.27-3.85; P = 0.80). CONCLUSIONS: Among women with HIV in a prospective study that incorporated cervical cancer prevention measures, the incidence of ICC was not significantly higher than that in a comparison group of HIV-negative women. PMID- 19127539 TI - Epigenetic control. AB - Epigenetics refers to mitotically and/or meiotically heritable variations in gene expression that are not caused by changes in DNA sequence. Epigenetic mechanisms regulate all biological processes from conception to death, including genome reprogramming during early embryogenesis and gametogenesis, cell differentiation and maintenance of a committed lineage. Key epigenetic players are DNA methylation and histone post-translational modifications, which interplay with each other, with regulatory proteins and with non-coding RNAs, to remodel chromatin into domains such as euchromatin, constitutive or facultative heterochromatin and to achieve nuclear compartmentalization. Besides epigenetic mechanisms such as imprinting, chromosome X inactivation or mitotic bookmarking which establish heritable states, other rapid and transient mechanisms, such as histone H3 phosphorylation, allow cells to respond and adapt to environmental stimuli. However, these epigenetic marks can also have long-term effects, for example in learning and memory formation or in cancer. Erroneous epigenetic marks are responsible for a whole gamut of diseases including diseases evident at birth or infancy or diseases becoming symptomatic later in life. Moreover, although epigenetic marks are deposited early in development, adaptations occurring through life can lead to diseases and cancer. With epigenetic marks being reversible, research has started to focus on epigenetic therapy which has had encouraging success. As we witness an explosion of knowledge in the field of epigenetics, we are forced to revisit our dogma. For example, recent studies challenge the idea that DNA methylation is irreversible. Further, research on Rett syndrome has revealed an unforeseen role for methyl-CpG-binding protein 2 (MeCP2) in neurons. PMID- 19127540 TI - Role of vascular endothelial growth factor in the communication between human osteoprogenitors and endothelial cells. AB - Proper bone remodeling requires an active process of angiogenesis which in turn supplies the necessary growth factors and stem cells. This tissue cooperation suggests a cross-talk between osteoblasts and endothelial cells. This work aims to identify the role of paracrine communication through vascular endothelial growth factor (VEGF) in co-culture between osteoblastic and endothelial cells. Through a well defined direct contact co-culture model between human osteoprogenitors (HOPs) and human umbilical vein endothelial cells (HUVECs), we observed that HUVECs were able to migrate along HOPs, inducing the formation of specific tubular-like structures. VEGF(165) gene expression was detected in the HOPs, was up-regulated in the co-cultured HOPs and both Flt-1 and KDR gene expression increased in co-cultured HUVECs. However, the cell rearrangement observed in co-culture was promoted by a combination of soluble chemoattractive factors and not by VEGF(165) alone. Despite having no observable effect on endothelial cell tubular-like formation, VEGF appeared to have a crucial role in osteoblastic differentiation since the inhibition of its receptors reduced the co culture-stimulated osteoblastic phenotype. This co-culture system appears to enhance both primary angiogenesis events and osteoblastic differentiation, thus allowing for the development of new strategies in vascularized bone tissue engineering. PMID- 19127541 TI - Wnt pathway, an essential role in bone regeneration. AB - Fracture repair is a complex regenerative process initiated in response to injury, resulting in optimal restoration of skeletal function. Although histology characteristics at various phases of fracture repair are clear and well established, much remains to be understood about the process of bone healing, particularly at the molecular signaling level. During the past decade, secreted signaling molecules of the Wnt family have been widely investigated and found to play a central role in controlling embryonic development processes. Wnt signaling pathway also plays a pivotal role in the regulation of bone mass. Recent published data reveal that Wnt signaling pathway is activated during postnatal bone regenerative events, such as ectopic endochondral bone formation and fracture repair. Dysregulation of this pathway greatly inhibits bone formation and healing process. Interestingly, activation of Wnt pathway has potential to improve bone healing, but only utilized after mesenchymal cells have become committed to the osteoblast lineage. These advances suggest an essential role of Wnt pathway in bone regeneration. PMID- 19127542 TI - Cloning and analysis of rat osteoclast inhibitory lectin gene promoter. AB - Osteoclast inhibitory lectin (OCIL) is a novel regulator of bone remodeling, however, little is known concerning how OCIL is regulated to date. In this study, approximately 4.4 kb of the 5'-flanking sequence of rat OCIL gene was cloned into the promoter-less reporter vector pGL3-basic and transiently transfected into three different cell lines. The differences in the levels of luciferase activity paralleled well with the levels of OCIL mRNA expression in these cells, suggesting that the regulation of rat OCIL gene expression occurs mainly at the transcriptional level. Additional luciferase assays using a series of constructs containing unidirectionally deleted fragments showed that the construct-1819/pGL3 (-1819 to +118) exhibited the highest luciferase activity, suggesting the presence of functional promoter in this region. The region from -4370 to -2805 might contain negative regulatory elements, while the region from -1819 to -1336 might have important positive regulatory elements that enhance OCIL transcription. Sequence analysis of the promoter revealed the absence of both TATA and CAAT boxes. However, in the proximal promoter region (-81 to +118), several potential transcription factor binding sites that may be responsible for the basal transcriptional activity of rat OCIL promoter were observed. The promoter contains several potential Sp1 binding sites, and cotransfection of a shRNA expression plasmid that knockdowns Sp1 significantly reduced OCIL promoter activity and endogenous gene expression and moreover, overexpressing Sp7, a Sp1 family member that also binds to Sp1 binding sequence, increased OCIL promoter activity and gene expression, suggesting a role of Sp1 family proteins in regulation of OCIL transcription. PMID- 19127543 TI - Increased ovarian cancer risk associated with menopausal estrogen therapy is reduced by adding a progestin. AB - BACKGROUND: It has become increasingly clear that use of menopausal hormone therapy (HT) is associated with an increased risk of ovarian cancer; however, the effects by type of formulation and duration of use are less clear. A systematic review of the HT and ovarian cancer literature was conducted to identify population-based case-control studies, cohort studies, and randomized trials that examined effects by formulation of HT (estrogen-alone [ET] and estrogen plus progestin [EPT]) and duration of use. METHODS: Pub-Med (www.pubmed.gov) was used to identify relevant publications through December 2007; 14 studies were identified. The authors abstracted relative risks (RRs) and 95% confidence intervals (CIs) in relation to duration of HT use (ET and EPT separately). The authors used the risk estimates per year of HT use if these were provided; otherwise, they calculated a duration-response for a log-linear model of the duration of HT use against risk. RESULTS: Ovarian cancer risk was increased among ET users (RR per 5 years of use, RR(5) = 1.22; 95% CI, 1.18-1.27; P < .0001), and a lower but still statistically significant increased risk was seen with EPT use (RR(5) = 1.10; 95% CI, 1.04-1.16; P = .001). The increased risk in ET users was statistically significantly higher than the increased risk in EPT users (P = .004). CONCLUSIONS: ET use increases risk of ovarian cancer in a duration dependent manner, and it appears that the addition of progestins blocks this effect, at least to some extent. Whether the effect of estrogens would be completely blocked if progestins were given every day is unclear. PMID- 19127544 TI - Diversity of participants in clinical trials in an academic medical center: the role of the 'Good Study Patient?'. AB - BACKGROUND: Only 2.5% of adults and even fewer minorities participate in cancer therapeutic trials. Researchers have concluded that many barriers to participation stem from how recruitment is performed by clinician investigators. The objective of the current research was to document specifically how these barriers impede recruitment in the clinical setting. METHODS: The authors conducted a case study of recruitment in an academic medical center using ethnographic research methods (direct observation of provider-patient interactions and in-depth interviews with providers) to collect data. Qualitative data analysis was used to identify themes related to the provider's role in the recruitment processes. RESULTS: In the clinics that were studied, the authors observed that providers subjectively assessed which patients seemed to be 'good study patients' to target for recruitment. 'Good study patients' were identified as those who were able to adhere to complex trial protocols, thus helping clinician researchers to complete studies in a timely and efficient manner. These patients were perceived as meticulous, proactive, and compliant; they were considered good communicators; and they were embedded in the kinds of strong social support networks that facilitated their trials participation. CONCLUSIONS: The providers that were studied sought 'good study patients' for therapeutic trials because they wanted to perform studies in a timely and efficient manner. Future research should examine whether providers in other settings also target their recruitment efforts for this or other reasons. Further research also should consider whether differentially recruiting 'good patients' can impact the ethnic/racial diversity or other characteristics of trial participants in ways that may bias the outcomes or conclusions of therapeutic trials. PMID- 19127545 TI - Parathyroid hormone regulation of the human bone sialoprotein gene transcription is mediated through two cAMP response elements. AB - Parathyroid hormone (PTH) regulates serum calcium and inorganic phosphate levels through its actions on kidney and bone. Bone sialoprotein (BSP) is an early marker of osteoblast differentiation and bone metabolism. We here report that two cAMP response elements (CRE) in the human BSP gene promoter are target of PTH. In human osteoblast-like Saos2 cells, PTH (human 1-34 PTH, 10 nM) increased BSP mRNA and protein levels at 3 h. From transient transfection assays, 2- to 2.5-fold increase in transcription by PTH was observed at 3 and 6 h in -184, -211, -428, 868, and -927 luciferase constructs that included the human BSP gene promoter. Effect of PTH was abrogated by 2 bp mutations in either the CRE1 (-79 to -72) or CRE2 (-674 to -667). Luciferase activities induced by PTH were blocked by protein kinase A inhibitor H89 and tyrosine kinase inhibitor herbimycin A. Gel shift analyses showed that PTH increased binding of nuclear proteins to the CRE1 and CRE2 elements. The CRE1-protein and CRE2-protein complexes were disrupted by CRE binding protein 1 (CREB1) antibodies and supershifted by phospho-CREB1 antibody. ChIP assays detected binding of CREB1 and phospho-CREB1 to a chromatin fragment containing CRE1 and CRE2, and increased binding of phospho-CREB1 to the both sites. These studies demonstrate that PTH stimulates human BSP gene transcription by targeting the two CREs in the promoter of the human BSP gene. PMID- 19127546 TI - Pharmaceutical metabolite profiling using quadrupole/ion mobility spectrometry/time-of-flight mass spectrometry. AB - The use of hybrid quadrupole ion mobility spectrometry time-of-flight mass spectrometry (Q/IMS/TOFMS) in the metabolite profiling of leflunomide (LEF) and acetaminophen (APAP) is presented. The IMS drift times (T(d)) of the drugs and their metabolites were determined in the IMS/TOFMS experiments and correlated with their exact monoisotopic masses and other in silico generated structural properties, such as connolly molecular area (CMA), connolly solvent-excluded volume (CSEV), principal moments of inertia along the X, Y and Z Cartesian coordinates (MI-X, MI-Y and MI-Z), inverse mobility and collision cross-section (CCS). The correlation of T(d) with these parameters is presented and discussed. IMS/TOF tandem mass spectrometry experiments (MS(2) and MS(3)) were successfully performed on the N-acetyl-p-benzoquinoneimine glutathione (NAPQI-GSH) adduct derived from the in vitro microsomal metabolism of APAP. As comparison, similar experiments were also performed using hybrid triple quadrupole linear ion trap mass spectrometry (QTRAPMS) and quadrupole time-of-flight mass spectrometry (QTOFMS). The abilities to resolve the product ions of the metabolite within the drift tube and fragment the ion mobility resolved product ions in the transfer travelling wave-enabled stacked ring ion guide (TWIG) demonstrated the potential applicability of the Q/IMS/TOFMS technique in pharmaceutical metabolite profiling. PMID- 19127547 TI - Liquid chromatography coupled with ultraviolet absorbance detection, electrospray ionization, collision-induced dissociation and tandem mass spectrometry on a triple quadrupole for the on-line characterization of polyphenols and methylxanthines in green coffee beans. AB - Liquid chromatography coupled with a photodiode array detector, electrospray ionization, collision-induced dissociation and tandem mass spectrometry (LC DAD/ESI-CID-MS/MS) on a triple quadrupole (QqQ) has been used to detect and characterize polyphenols and methylxanthines in green coffee beans: three phenolic acids (caffeic acid, ferulic acid and dimethoxycinnamic acid), three isomeric caffeoylquinic acids (M(r) 354), three feruloylquinic acids (M(r) 368), one p-coumaroylquinic acid (M(r) 338), three dicaffeoylquinic acids (M(r) 516), three feruloyl-caffeoylquinic acids (M(r) 530), four p-coumaroyl-caffeoylquinic acids (M(r) 500), three diferuloylquinic acids (M(r) 544), six dimethoxycinnamoyl caffeoylquinic acids (M(r) 544), three dimethoxycinnamoyl-feruloylquinic acids (M(r) 558), six cinnamoyl-amino acid conjugates, three cinnamoyl glycosides, and three methylxanthines (caffeine, theobromine and theophylline). Dimethoxycinnamic acid, three isomers of dimethoxycinnamoyl-caffeoylquinic acids and another three of dimethoxycinnamoyl-feruloylquinic acids, as well as the three cinnamoyl glycosides, had not previously been reported in coffee beans. Structures have been assigned on the basis of the complementary information obtained from UV visible spectra, relative hydrophobicity, scan mode MS spectra, and fragmentation patterns in MS(2) spectra (both in the positive and negative ion modes) obtained using a QqQ at different collision energies. A structure diagnosis scheme is provided for the identification of different isomers of polyphenols and methylxanthines. PMID- 19127548 TI - The proteolytic activity of insulin-degrading enzyme: a mass spectrometry study. AB - The prominent role that insulin-degrading enzyme (IDE) has on amyloidogenic peptides degradation has recently boosted a lot of attention toward this enzyme. Although many substrates are known to be degraded by IDE, little is known about the changes in the proteolytic activity of the enzyme upon modification of environmental factors. In a previous work we have already shown the great potentiality of atmospheric pressure/laser desorption ionization-mass spectrometry (AP/MALDI-MS) for studying the interaction between IDE and insulin. Here, the activity of IDE was investigated regarding cleavage sites' preferentiality upon modification of environmental factors by AP/MALDI-MS. The roles that IDE/insulin concentration ratio, reaction time, adenosine 5' triphosphate (ATP) and metal ions (Zn and Cu) have on the insulin cleavage pattern produced by IDE are investigated and a plausible interpretation involving the proteolytic action of the different IDE oligomeric forms is proposed. PMID- 19127549 TI - Determination of trichloroethylene from adipose tissue by headspace solid-phase microextraction gas chromatography/mass spectrometry. PMID- 19127550 TI - Comparing auditory perception and speech production outcomes: non-language specific assessment of auditory perception and speech production in children with cochlear implants. AB - Language-independent assessment tools evaluate the progress of children who receive a cochlear implant, allowing large pooling of data for better access to insurers and other health care professionals. One hundred and seventeen children from centres in the United Kingdom, Iran and Turkey were assessed on two measures over a five-year test interval. There is a significant improvement over time for the Categories of Auditory Performance (CAP) and Speech Intelligibility Rating (SIR) measures. There was a significant difference between scores for different language groups: accounted for by the differences in age at implantation. There was a significant effect of age at implantation up to three years of device use. There were high correlations between the CAP and SIR scores. A longer duration of deafness resulted in a higher score for both scales; however, there was no relationship when correlated for age. Finally, the CAP pre-operative score allows us to predict the post-operative SIR scores. The scales are validated; reliable measures which can be used across countries and languages. This allows greater ability to pool data allowing data to be generalised across population groups, providing more power to prove that cochlear implantation is a viable treatment for children with bilateral severe-to-profound hearing loss. PMID- 19127551 TI - Suppressing one-bond correlations in HMBC spectra: improved performance for the BIRD-HMBC pulse sequence. AB - An improved version of the BIRD-HMBC experiment is proposed. In comparison to the original version, the filtering (suppression of (1) J(CH) signals) is accomplished using a double tuned G-BIRD filter positioned in the middle of the long-range correlations evolution period. Compensation of offset dependence by replacing the rectangular 180 degree pulses with the broadband inversion pulses (BIPs), with superior inversion performance and improved tolerance to B(1) field inhomogeneity, significantly improves the sensitivity of the original BIRD-HMBC experiment. For usual one-bond coupling constants ranges (115-180 Hz), optimal results are easily obtained by adjusting the delays, delta, of the BIRD elements to an average J value. For larger ranges (e.g. 110-260 Hz), the use of a double tuned G-BIRD filter allows excellent suppression degrees for all types of one bond constants present in a molecule, superior to the original scheme and other purging schemes. These attributes make the improved version of the BIRD-HMBC experiment a valuable and robust tool for rapid spectral analysis and rapid checks of molecular skeletons with a minimum spectrometer time. PMID- 19127552 TI - Body mass index and risk of ovarian cancer. AB - BACKGROUND: Convincing epidemiologic evidence links excess body mass to increased risks of endometrial and postmenopausal breast cancers, but the relation between body mass index (BMI) and ovarian cancer risk remains inconclusive. Potential similarities regarding a hormonal mechanism in the etiology of female cancers highlight the importance of investigating associations according to menopausal hormone therapy (MHT) use. However, to the authors' knowledge, data addressing whether the relation between BMI and ovarian cancer differs by MHT use are very sparse. METHODS: The authors prospectively investigated the association between BMI and ovarian cancer among 94,525 US women who were followed between 1996 through 1997 to December 31, 2003. During 7 years of follow-up, 303 epithelial ovarian cancer cases were documented. RESULTS: Compared with normal weight women (BMI of 18.5-24.9 kg/m(2)), the multivariate relative risk (MVRR) of ovarian cancer for obese women (BMI of >or=30 kg/m(2)) in the cohort as a whole was 1.26 (95% confidence interval [95% CI], 0.94-1.68). Among women who never used MHT, the MVRR for obese versus normal weight women was 1.83 (95% CI, 1.18-2.84). In contrast, no relation between BMI and ovarian cancer was apparent among women who ever used MHT (MVRR = 0.96; 95% CI, 0.65-1.43; P interaction = 0.02). Exploratory analyses also suggested a positive association between BMI and ovarian cancer among women without a family history of ovarian cancer (MVRR comparing obese vs normal weight women = 1.36; 95% CI, 1.00-1.86), but no relation with BMI was apparent among women with a positive family history of ovarian cancer (MVRR = 0.74; 95% CI, 0.34-1.62 [P interaction = .02]). CONCLUSIONS: Based on the results of the current study, the authors suspect that obesity is associated with enhanced ovarian cancer risk through a hormonal mechanism. PMID- 19127555 TI - The patient behind the code, the care behind the patient. PMID- 19127556 TI - Accuracy of the BRCAPRO model among women with bilateral breast cancer. AB - BACKGROUND: The likelihood of identifying a BRCA mutation was often calculated using the BRCAPRO model. A previous study suggested that this model may overestimate the chance of detecting a BRCA mutation among women diagnosed with bilateral breast cancer. Studies also suggested that few patients with bilateral breast cancer whose age at first diagnosis is >40 years were mutation carriers. The objectives of this study were to determine the accuracy of the BRCAPRO model among women with bilateral breast cancer and to determine whether their mutation status was dependent on their age at first diagnosis. METHODS: A retrospective chart review was performed. Women who were diagnosed with bilateral or unilateral breast cancer and who had undergone comprehensive BRCA1 and BRCA2 genetic testing at M. D. Anderson Cancer Center between 1997 and 2006 were included in the study. RESULTS: For individuals with pre-test carrier probabilities >31%, the proportion of positive tests was significantly lower than predicted by the BRCAPRO model (P < .05). In addition, the carrier rate of BRCA mutations was significantly higher (P = .002, Fisher exact test) in women with bilateral breast cancer whose age at first diagnosis was 40 years. CONCLUSIONS: The BRCAPRO model was overestimating the relative contribution bilateral breast cancer had on the likelihood of detecting a BRCA1 or BRCA2 mutation. Bilateral breast cancer did not appear to be a good indicator of mutation status, particularly for women whose age at first diagnosis is >40 years. PMID- 19127558 TI - Invader human papillomavirus (HPV) type 16 and 18 assays as adjuncts to HPV screening of cervical papanicolaou smears with atypical squamous cells of undetermined significance. AB - BACKGROUND: High-risk (HR) human papillomavirus (HPV) testing is standard practice for triaging women who have Papanicolaou (Pap) smears with atypical squamous cells of undetermined significance (ASC-US), however, only 5% to 17% of these women have underlying cervical intraepithelial neoplasia 2 (CIN-2)/CIN-3. Recent reports have demonstrated that the presence of either HPV type 16 (HPV-16) or HPV-18 confers an elevated risk for CIN-2/CIN-3. The current study was designed to determine the prevalence of HPV-16 and HPV-18 in ASC-US Pap smears and to determine whether further typing would enhance the risk stratification of patients for CIN-2/CIN-3. METHODS: One hundred seventy-eight Pap smears with ASC US were screened retrospectively for HR HPV by using the proprietary Invader screening assay followed by typing for HPV-16 and HPV-18 by using Invader type specific probes on 100 of the samples. Clinical follow-up results were correlated with HPV types. RESULTS: Fifty-one percent of the ASC-US samples were positive for HR HPV, the majority of which (70%) harbored non-HPV-16/HPV-18 HR HPV types; 27% were associated with HPV-16, whereas only 3% contained HPV-18. The screening assay indicated that 46% of women who had Pap smears with ASC-US were in need of further HPV-16/HPV-18 typing. Testing for HPV-16 stratified women with ASC-US into 3 groups: 1) 14% of women were positive for HPV-16 and had a high risk (54%) of CIN-2/CIN-3 on follow-up biopsy, 2) 35% of women were positive for non-HPV-16 HPV types and had an intermediate risk (9%), and 3) 51% of women were negative for HPV and had a negligible risk for CIN-2/CIN-3. CONCLUSIONS: The combined application of a proprietary screening assay and a type-specific HPV-16 assay demonstrated global potential for the development of tailored management protocols for women who have Pap smears with ASC-US. PMID- 19127557 TI - Lack of pathologic down-staging with neoadjuvant chemotherapy for muscle-invasive urothelial carcinoma of the bladder: a contemporary series. AB - BACKGROUND: The postcystectomy survival benefit associated with the combination of methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) neoadjuvant chemotherapy (NC) for muscle-invasive bladder cancer has been most evident in patients who achieve a pathologic complete response. The outcome of NC and open radical cystectomy (RC) was evaluated in a contemporary cohort of patients in a tertiary referral setting. METHODS: From January 2006 to November 2007, 117 patients underwent open RC at Cleveland Clinic for muscle-invasive bladder cancer, 29 (25%) of whom received NC. Patient information was obtained from a prospective database. RESULTS: Clinical stage at the time of diagnosis in the NC cohort was T2 in 23 (79%) and T3-4a in 6 (21%) patients. A total of 20 (69%) patients received the combination of gemcitabine and cisplatin (GC), 4 (14%) received MVAC, and 5 (17%) received other regimens. The median interval from the time of diagnosis of muscle-invasive bladder cancer to RC was 208 days (interquartile range, 149 days -327 days) in the NC cohort. Overall, only 2 patients (7%; 95% confidence interval [95% CI], 0 patients-17 patients) achieved a pathologic complete response, 18 (62%; 95% CI, 43 patients-81 patients) had nonorgan-confined residual cancer, and the overall median progression-free survival was 10.5 months (95% CI, 7 months -14 months). CONCLUSIONS: Few RC patients in these investigators' recent experience achieved a pathologic complete response with NC, and most experienced rapid disease progression. These poor outcomes may be related to the use of non-MVAC-based regimens or excessive delay in performing RC. In the absence of supportive data for GC in the neoadjuvant setting, MVAC remained the preferred regimen. Excessive delays in performing RC may negate the benefit of NC. PMID- 19127560 TI - A population-based study evaluating the impact of sunitinib on overall survival in the treatment of patients with metastatic renal cell cancer. AB - BACKGROUND: Sunitinib has replaced interferon (IFN) as a first-line standard of care in the treatment of metastatic renal cell carcinoma (RCC). This study aimed to determine overall survival and to confirm effectiveness in a population that includes poor prognosis patients. METHODS: Data were collected on all patients identified by the BC Cancer Registry with metastatic RCC who were treated with IFN or sunitinib. The IFN group consisted of patients who received IFN between January 2000 and October 2005, and the sunitinib group included patients treated with first-line sunitinib from October 2005 to September 2007. RESULTS: There were 131 and 69 patients in the IFN and sunitinib groups, respectively. The median follow-up of those still alive was 12.6 months. The median age (62 vs 63 years; P = .41), Memorial Sloan Kettering Cancer Center (MSKCC) prognostic criteria (poor in 19% vs 30%; P = .41), and proportion with >1 metastasis (53% vs 62%; P = .21) were similar between the IFN and sunitinib groups, respectively. The median survival of the IFN and sunitinib groups was 8.7 and 17.3 months, respectively (log-rank P = .004). The median survival of patients with favorable, intermediate, and poor MSKCC prognostic profiles in the IFN group was 22.9, 8.7, and 4.1 months, respectively (P < .001), whereas in the sunitinib group it was not reached, 16.8, and 10.7 months, respectively (P = .006). The hazard ratio of death after adjusting for MSKCC criteria was 0.49 (95% confidence interval, 0.31 0.76; P = .001). CONCLUSIONS: The introduction of first-line sunitinib was associated with a doubling of overall survival compared with patients treated with IFN alone. This benefit extended to patients with poor MSKCC prognostic profiles. PMID- 19127561 TI - HiRes with Fidelity 120 benefit in native speakers of Korean. AB - In HiRes 120 sound processing, the spectral bands are created by precisely varying the proportion of current delivered simultaneously to adjacent electrodes through active current steering. The purpose of this study was to examine performance of native Korean speakers with HiRes 120. Eleven adults with postlinguial hearing loss participated in the study. The difference between baseline and three-month HiRes 120 performance was significant for all tests (p < 0.05). For monosyllabic words, eight subjects obtained higher scores with HiRes 120. For the Hearing in Noise Test (HINT), ten subjects obtained higher scores with standard HiRes. These results show Korean speakers exhibit improved speech understanding in quiet and in noise with HiRes 120. All subjects preferred the new HiRes 120 sound processing option. PMID- 19127559 TI - Microsatellite instability, MLH1 promoter methylation, and BRAF mutation analysis in sporadic colorectal cancers of different ethnic groups in Israel. AB - BACKGROUND: The molecular mechanisms that underlie colorectal cancer (CRC) include microsatellite instability (MSI), chromosomal instability, and the CpG island methylator phenotype. There is evidence to suggest that CRC incidence varies among different ethnic populations worldwide. The authors of this report hypothesized that environmental factors and lifestyle differences among various ethnic groups may differentially influence the epigenetic regulation of tumor suppressor genes in CRC. METHODS: In the current study, microdissection and DNA extraction were performed on 128 samples of CRC from Israeli patients (85 Jews and 43 Arabs). MSI analysis, mutL homolog 1 (MLH1) and mutS homolog 2 (MSH2) protein expression levels, and MLH1 promoter methylation were investigated by combined bisulfite restriction analysis. The v-raf murine sarcoma viral oncogene homolog B1 (BRAF) valine-to-glutamic acid mutation at residue 600 was investigated by direct DNA sequencing. RESULTS: High MSI (MSI-H), MLH1 methylation, and BRAF mutations were observed in 11.6%, 9.4%, and 23.5% of Jews, respectively, and in 16.2%, 17.6%, and 20.9% of Arabs, respectively (P value nonsignificant). MLH1 promoter methylation was observed in 22.6% of microsatellite-stable (MSS) tumors and in 53.8% of MSI-H tumors (P < .015). Extensive methylation (covering both 5' and 3' promoter regions) was present in all MSI-H tumors with loss of MLH1 expression. BRAF mutation was observed in 15.6% and 46.1% of MSS tumors and MSI-H tumors, respectively (P < .007). BRAF mutation was observed in 66%, 22.2%, and 14.7% of patients who had tumors with extensive MLH1 promoter methylation, methylation of the 5' region alone, or without methylation, respectively (P < .006). CONCLUSIONS: There was no difference in molecular signatures examined between Jewish and Arab patients with CRC in Israel. Extensive promoter methylation was associated with MLH1 inactivation, MSI, and BRAF mutation. PMID- 19127562 TI - The multi-channel cochlear implant: past, present and future perspectives. AB - Initial research demonstrated that only low frequencies could be mimicked with rate of electrical stimulation, and thus multi-channel rather than single-channel stimulation was required for the place coding of the mid-high speech frequencies. Place coding of mid-high frequencies was best achieved with electrodes inside the cochlea. Furthermore, correct biomechanical properties of a multiple electrode bundle were required for it to pass around the cochlear spiral to the speech frequency region. Biological studies showed too that intra-cochlear electrodes could be used with minimal trauma, safe electrical stimulus parameters, and methods to prevent inner ear infection and meningitis. The crucial discoveries for coding speech with electrical stimulation have been based on the discovery of: 1) the fact the brain processes frequency information along spatial and temporal channels, and 2) that the first patient experienced vowels when stimulating different electrodes that corresponded to the place of excitation for single formant vowels in people with normal hearing. The inaugural and subsequent speech processing strategies extracted frequencies of special importance for speech intelligibility, and transmitted the information along place coding channels. The voicing frequency and/or amplitude, was coded as temporal information across these spatial channels. As a result a great majority of severely-to-profoundly deaf people with previous hearing can not only communicate when electrical stimulation is combined with lipreading, but with electrical stimulation alone. In addition, the benefits of binaural hearing with bilateral cochlear implants or an implant in one ear and hearing aid in the other ear have been realized. Related psychophysical research has discovered the basic perceptual skills that process the complex patterns of brain excitation that underlie speech recognition both in the one ear as well as bilateral implants.In addition the development of the perceptual skills in the maturing child for speech recognition, have been discovered. In the future high fidelity sound should be achieved by providing the fine temporo-spatial patterns of excitation and preserving the peripheral nerve network. This could require the release of nerve growth factors and the development of electrodes using nanotechnology. PMID- 19127564 TI - Convergent genome wide association results for bipolar disorder and substance dependence. AB - Twin studies document substantial heritability for substance dependence and bipolar disorder [Shih et al. (2004); Uhl et al. (2008a)]. Individuals with bipolar disorder display substance use disorders at rates that are much higher than those in the general population [Krishnan (2005)]. We would thus predict: 1) substantial overlap between different genome wide association (GWA) studies of bipolar disorder 2) significant overlap between results from bipolar disorder and substance dependence. Recent GWA studies [Baum et al. (2007); Sklar et al. (2008); Uhl et al. (2008a); Wellcome Trust Consortium (2007)] allow us to test these ideas, although 1) these datasets display difficult features that include use of differing sets of SNPs, likely polygenic genetics, likely differences in linkage disequilibrium between samples, heterogeneity both between and within loci and 2) several, though not all, reports have failed to identify any allele of any single nucleotide polymorphism (SNP) ("same SNP same allele") that is reproducibly associated with bipolar disorder with "genome wide" significance. We now report analyses that identify clustered, P < 0.05 SNPs within genes that overlap between the bipolar samples (Monte Carlo P < 0.00001). Overlapping data from at least three of these studies identify 69 genes. 23 of these genes also contain overlapping clusters of nominally-positive SNPs for substance dependence. Variants in these "addiction/bipolar" genes are candidates to influence the brain in ways that manifest as enhanced vulnerabilites to both substance dependence and bipolar disorder. PMID- 19127565 TI - Deletion of BMP7 affects the development of bones, teeth, and other ectodermal appendages of the orofacial complex. AB - Sequential and reciprocal epithelial-mesenchymal interactions govern the development of most tissues and organs of the craniofacial region. Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-beta family of secreted signaling molecules that have long been implied to have a significant contribution in this process. However, evidence for such a role during craniofacial development is largely missing. Using a lacZ reporter mouse we mapped the spatiotemporal expression of BMP7 in the developing craniofacial region. The observed pattern suggested a potential involvement of BMP7 in epithelial-mesenchymal interactions and thus a direct role for this molecule in the development of ectodermal appendages (teeth, hair follicle, lachrymal and sweat glands, taste buds) and, furthermore, palatal formation. To correlate the expression to function we analyzed germline deleted conditional BMP7-deficient embryos for malformations. We found developmental defects in many craniofacial structures such as teeth, eyes, whiskers, hair follicles, salivary glands, and palate. These findings place BMP7 as a central mediator of epithelial-mesenchymal interactions that are necessary for the correct development of structures belonging to the orofacial complex. PMID- 19127563 TI - Family-based association study of Neuregulin 1 with psychotic bipolar disorder. AB - The Neuregulin 1 gene (NRG1) has been associated with schizophrenia, and, to a lesser extent, with bipolar disorder (BP). We investigated the association of NRG1 with BP in a large family sample, and then performed analyses according to the presence of psychotic features or mood-incongruent psychotic features. We genotyped 116 tagSNPs and four Icelandic "core" SNPs in 1,199 subjects from 314 nuclear families. Of 515 BP offspring, 341 had psychotic features, and 103 had mood-incongruent psychotic features. In single-marker and sliding window haplotype analyses using FBAT, there was little association using the standard BP or mood-incongruent psychotic BP phenotypes, but stronger signals were seen in the psychotic BP phenotype. The most significant associations with psychotic BP were in haplotypes within the 5' "core" region. The strongest global P-value was across three SNPs: NRG241930-NRG243177-rs7819063 (P = 0.0016), with an undertransmitted haplotype showing an individual P = 0.0007. The most significant individual haplotype was an undertransmitted two-allele subset of the above (NRG243177-rs7819063, P = 0.0004). Additional associations with psychotic BP were found across six SNPs in a 270 kb central region of the gene. The most 3' of these, rs7005606 (P = 0.0029), is located approximately 4 kb from the type I NRG1 isoform promoter. In sum, our study suggests that NRG1 may be specifically associated with the psychotic subset of BP; however, our results should be interpreted cautiously since they do not meet correction for multiple testing and await independent replication. PMID- 19127566 TI - Toward an "omic" physiopathology of reactive chemicals: thirty years of mass spectrometric study of the protein adducts with endogenous and xenobiotic compounds. AB - Cancer and degenerative diseases are major causes of morbidity and death, derived from the permanent modification of key biopolymers such as DNA and regulatory proteins by usually smaller, reactive molecules, present in the environment or generated from endogenous and xenobiotic components by the body's own biochemical mechanisms (molecular adducts). In particular, protein adducts with organic electrophiles have been studied for more than 30 [see, e.g., Calleman et al., 1978] years essentially for three purposes: (a) as passive monitors of the mean level of individual exposure to specific chemicals, either endogenously present in the human body or to which the subject is exposed through food or environmental contamination; (b) as quantitative indicators of the mean extent of the individual metabolic processing which converts a non-reactive chemical substance into its toxic products able to damage DNA (en route to cancer induction through genotoxic mechanisms) or key proteins (as in the case of several drugs, pesticides or otherwise biologically active substances); (c) to relate the extent of protein modification to that of biological function impairment (such as enzyme inhibition) finally causing the specific health damage. This review describes the role that contemporary mass spectrometry-based approaches employed in the qualitative and quantitative study of protein electrophile adducts play in the discovery of the (bio)chemical mechanisms of toxic substances and highlights the future directions of research in this field. A particular emphasis is given to the measurement of often high levels of the protein adducts of several industrial and environmental pollutants in unexposed human populations, a phenomenon which highlights the possibility that a number of small organic molecules are generated in the human organism through minor metabolic processes, the imbalance of which may be the cause of "spontaneous" cases of cancer and of other degenerative diseases of still uncharacterized etiology. With all this in mind, it is foreseen that a holistic description of cellular functions will take advantage of new analytical methods based on time integrated metabolomic measurements of a new biological compartment, the "adductome," aimed at better understanding integrated organism response to environmental and endogenous stressors. PMID- 19127567 TI - Treatment strategies and regimens of graduated intensity for childhood acute lymphoblastic leukemia in low-income countries: A proposal. AB - Cure rates for children with acute lymphoblastic leukemia (ALL) are 80-85% in high-income countries (HICs) in North America and Western Europe. However, cure rates are much lower in many low-income countries (LICs), where most cases of ALL occur. Over the past several decades partnerships ("twinning") between HIC and LIC pediatric oncology programs have led to major improvements in outcome for children with ALL in some LICs, often by developing time and resource intensive relationships that allow LIC centers to treat children with regimens similar or identical to those used in HICs. However, the resources are not available in most LICs to allow immediate introduction of intensive ALL treatment regimens similar to those used in HICs. With these thoughts in mind, we present a proposal for a systematic and graduated approach to ALL diagnosis, risk classification, and treatment in LICs. We have based the strategy and the proposed regimens on those developed by the Children's Cancer Group (CCG) and Children's Oncology Group (COG) over the past several decades, beginning with a first level regimen similar to CCG therapy of the early 1980s and then layering on successive treatment intensifications proven effective in randomized clinical trials. Simple monitoring rules are included to help centers decide when they are ready to add new treatment components. This proposal provides a framework that LIC centers can use to provide effective ALL therapy, particularly in regions of the world where few children are currently being cured. PMID- 19127568 TI - Severe Evans syndrome with multi-system involvement is a distinct immunodeficiency disorder. AB - A female infant who presented with autoimmune hemolytic anemia and thrombocytopenia subsequently developed hepatic, dermatologic, renal, pulmonary, gastrointestinal, endocrine, and nervous system involvement. Prolonged and intensive treatment with prednisone, IVIG, mycophenolate mofetil, and anti-CD20 and anti-CD52 antibodies was necessary to control the symptoms. Laboratory evaluation showed normal lymphocyte subsets and function. There was normal Foxp3 and CD25 expression, no increased CD4(-)CD8(-) T-cell population, and the AIRE and Fas genes were without mutations. These features place the patient at the most severe portion of the Evans syndrome spectrum, and suggest that this case may represent a rare, new immunodeficiency disorder. PMID- 19127569 TI - Abnormal NT-pro-BNP levels in asymptomatic long-term survivors of childhood cancer treated with anthracyclines. AB - BACKGROUND: Anthracycline-induced cardiotoxicity can cause serious health problems for an increasing number of survivors of childhood malignancies. The aims of this study were to document plasma concentrations of cardiac troponin T (cTnT) and NT-pro-brain natriuretic peptide (NT-pro-BNP) in a large group of asymptomatic long-term survivors of childhood cancer treated with anthracyclines, and to study the relation of the abnormal biomarker levels with different risk factors for anthracycline-induced cardiotoxicity and conventional echocardiographic parameters. PROCEDURES: One hundred twenty-two asymptomatic survivors of childhood cancer underwent a detailed echocardiography. Blood samples were taken to determine the levels of NT-pro-BNP and cTnT. RESULTS: None of the survivors had abnormal cTnT levels. Thirteen percent of the survivors (n = 16) had abnormal NT-pro-BNP levels. Abnormal NT-pro-BNP levels were significantly related to cumulative anthracycline dosage (P < 0.003). Eleven of 31 survivors (35%) treated with cumulative anthracycline dose of 300 mg/m(2) or more, had abnormal NT-pro-BNP levels which were significantly related to end-diastolic left ventricular internal diameter (LVIDd) indexed for body surface area (BSA) (P < 0.01). CONCLUSION: Cardiac TnT does not contribute to the early detection of late onset anthracycline-induced cardiotoxicity. Abnormal levels of NT-pro-BNP were detected in 13% of 122 asymptomatic, long-term survivors of childhood cancer. Follow-up of these survivors is essential to answer the question whether NT-pro BNP is an early marker for late onset anthracycline-induced cardiotoxicity. PMID- 19127570 TI - Treatment of intestinal and hepatic mucormycosis in an immunocompromized child. AB - During ALL chemotherapy, a 4-year-old patient presented with febrile neutropenia and abdominal pain. Ultrasound examinations were repeatedly normal. Computerized tomography on day 7 demonstrated appendicitis and multiple hepatic foci identified as mucormycosis (Absidia corymbifera). Successful outcome was achieved by aggressive re-surgery, long-term antifungal therapy with serum level-monitored posaconazole, and recovery of neutrophil counts. Considering the interference of posaconazole with CYP3A4, vincristine was administered during 72 hr posaconazole windows. Pediatric intestinal mucormycosis, still associated with a >70% case fatality rate, calls for early imaging and surgery to establish the diagnosis, reduce the fungal mass, and provide a rationale for using posaconazole. PMID- 19127571 TI - Sickle cell disease and plumbism in children. AB - Childhood lead poisoning is common, affects multiple organs and is preventable. Co-existence of plumbism and sickle cell disease (SCD) is uncommon, with major overlap in signs and symptoms. We describe a patient with SCD and plumbism. Dysfunctional eating patterns may cause ingestion and increased absorption of lead in these patients. Routine screening for pica and other dysfunctional eating habits with appropriate behavioral interventions is emphasized. Annual screening for lead in patients with SCD of all ages (1-21years) and supplementation of micronutrients like zinc and iron in deficient cases may help prevent plumbism in these patients. PMID- 19127572 TI - Ovarian tumor in a 12-year old female with severe hypothyroidism: A case of Van Wyk and Grumbach syndrome. AB - We report a 12-year-old female presenting with an abdominal tumor. Diagnostic workup revealed giant bilateral ovarian cysts, severe hypothyroidism as well as an elevation of CA 125. We refrained from ovariectomy, which would be necessary for a malignant tumor, in view of an evident Van Wyk and Grumbach syndrome. The patient promptly responded to L-thyroxine with complete regression of all symptoms. Hypothyroidism should be considered in the evaluation of ovarian cysts. Although the Van Wyk and Grumbach syndrome is rare, it is crucial to rule it out in order to avoid unnecessary ovarian surgery when thyroid replacement is completely sufficient. PMID- 19127573 TI - Dopamine and impulse control disorders in Parkinson's disease. AB - There is an increasing awareness that impulse control disorders (ICDs), including compulsive gambling, buying, sexual behavior, and eating, can occur as a complication of Parkinson's disease (PD). In addition, other impulsive or compulsive disorders have been reported to occur, including dopamine dysregulation syndrome (DDS) and punding. Case reporting and prospective studies have reported an association between ICDs and the use of dopamine agonists (DAs), particularly at greater dosages, whereas dopamine dysregulation syndrome has been associated with greater dosages of levodopa or short-acting DAs. Data suggest that risk factors for an ICD may include male sex, younger age or younger age at PD onset, a pre-PD history of ICD symptoms, personal or family history of substance abuse or bipolar disorder, and a personality style characterized by impulsiveness. Although psychiatric medications are used clinically in the treatment of ICDs, there is no empiric evidence supporting their use in PD. Therefore, management for clinically significant ICD symptoms should consist of modifications to dopamine replacement therapy, particularly DAs, and there is emerging evidence that such management is associated with an overall improvement in ICD symptomatology. It is important that PD patients be aware that DA use may lead to the development of an ICD, and that clinicians monitor patients as part of routine clinical care. As empirically validated treatments for ICDs are emerging, it will be important to examine their efficacy and tolerability in individuals with cooccurring PD and ICDs. PMID- 19127574 TI - Parkinson's disease: unresolved issues. PMID- 19127575 TI - Elements of morphology: introduction. AB - An international group of clinicians working in the field of dysmorphology has initiated the standardization of terms used to describe human morphology. The goals are to standardize these terms and reach consensus regarding their definitions. In this way, we will increase the utility of descriptions of the human phenotype and facilitate reliable comparisons of findings among patients. Discussions with other workers in dysmorphology and related fields, such as developmental biology and molecular genetics, will become more precise. Here we describe the general background of the project and the various issues we have tried to take into account in defining the terms. PMID- 19127576 TI - Amyotrophic lateral sclerosis and neurosarcoidosis: a case report. AB - Amyotrophic lateral sclerosis (ALS) remains a clinical diagnosis without definable biomarkers. The pathomechanism of motor neuron degeneration in ALS has yet to be elucidated. Here we present a case of limb-onset ALS, with autopsy findings of Bunina bodies and skein-like inclusions, as well as sarcoid granulomas predominating among motor neurons. The targeting of the motor neurons by the sarcoid inflammation raises questions regarding the role of cellular immunity in the pathomechanisms for ALS. PMID- 19127577 TI - How do you treat motor complications in Parkinson's disease: Medicine, surgery, or both? AB - The motor complications associated with levodopa therapy, namely, fluctuations in motor response and dyskinesias, occur in the majority of Parkinson's disease patients. These complications can impair a patient's quality of life and even cause pronounced disability. "Off" states that result in freezing of gait and falling are disabling for many patients. Dyskinesias most commonly occur at peak dose and typically alternate with the wearing-off state. Once these problems appear, they usually persist, and the physician needs to make continual adjustments in medications to minimize these problems. Medical treatments should be attempted before treatments such as deep brain stimulation are considered because of the potential adverse effects that are associated with surgery. The timing of surgery, however, is also important because younger patients and less advanced patients tend to have a better outcome. There is thus a need for experienced and knowledgeable physicians and surgeons who are able to handle these motor complications. This review discusses available medications and surgical approaches, and their outcomes. PMID- 19127578 TI - Parkinson's disease dementia: definitions, guidelines, and research perspectives in diagnosis. AB - Cognitive impairment is common in Parkinson's disease (PD) and involves attentional, executive, visuospatial, and memory dysfunctions. Dementia is more frequently encountered in PD than in age-matched control populations, and whereas operational definitions of Alzheimer's disease and dementia with Lewy bodies have been developed, Parkinson's disease dementia (PD-D) has remained undefined. The Movement Disorder Society developed a task force to define and develop diagnostic guidelines for PD-D. This effort was based on existing descriptive studies with special emphasis on drawing distinction among Alzheimer's disease, dementia with Lewy bodies and PD-related cognitive impairment without dementia whenever possible. The second goal was to provide practical diagnostic procedures to diagnose PD-D. This effort emphasized available bedside tools that do not require neuropsychological expertise to administer or interpret. This work recently has been completed, and two primary articles have been published. The suggested clinical diagnostic criteria for PD-D involve four domains and are anchored in core features, associated clinical features, features that make the diagnosis uncertain, and features that are not compatible with the diagnosis of PD-D. When all four criteria are satisfactorily met, probable PD-D is designated; when the first and last criteria are met, but clinical characteristics are atypical or uncertainty factors exist, possible PD-D is designated. Whereas these definitions are operative and subject to change based on future data, they are based on widely available tests. The inclusion criteria can be applied internationally and in multicenter research on treatment interventions, clinicopathological correlations, and studies of cognitive and other nonmotor elements of PD. PMID- 19127579 TI - Drug selection and timing of initiation of treatment in early Parkinson's disease. AB - There is increasing evidence to challenge the traditional view that the initiation of drug treatment in Parkinson's disease (PD) should be delayed until the patient has significant disability such as to affect work or social function. Firstly, to delay treatment sentences the patient to protracted impairment of quality of life that could be improved by therapy. Secondly, there is evidence to support the notion that earlier rather than later initiation of treatment leads to better long term motor benefit. The selection of which drug to begin must be tailored to the patient's individual characteristics and circumstances. Monoamine oxidase B inhibitors result in a mild improvement in motor function compared to dopamine agonists or levodopa. They are well tolerated, easy to use once a day drugs and there is evidence that early use of Rasagiline improves motor outcome. Dopamine agonists lead to a substantial improvement in motor function and are, or will shortly be, available as once a day drugs. They are generally well tolerated but can be associated with exacerbating confusion or hallucinations and with behavioral changes. Levodopa is the most potent of the dopaminergic drugs. It is routinely combined with a dopa decarboxylase inhibitor and can also be used with a catecholo-o-methyl transferase inhibitor for enhanced absorption. The most important limiting factor for the use of levodopa is the emergence of motor complications. These are related to a number of factors including the dose of levodopa and the duration of its use. PMID- 19127580 TI - Why have we failed to achieve neuroprotection in Parkinson's disease? AB - The development of a neuroprotective therapy that slows, stops, or reverses neurodegeneration in Parkinson's disease (PD) is the single most important unresolved issue in the management of this disorder. Current therapies provide effective control of symptoms, particularly in the early stages of the disease, but disease progression is associated with the development of "nondopaminergic" features such as postural instability, falling, and dementia that are not adequately controlled with existing medications. There are many promising candidate neuroprotective agents based on pathological and laboratory studies, but to date, it has not been possible to determine that any drug has a disease modifying effect in PD. Obstacles to the development of a neuroprotective therapy in PD include: (1) uncertainty as to the precise cause of cell death in PD and what to target; (2) the lack of an animal model of PD that precisely reflects the etiopathogenesis of the disease, the pattern of dopaminergic and nondopaminergic pathology, and its chronic, progressive nature; (3) determination of the correct dose to use in clinical trials; and (4) delineation of a clinical end point that is an accurate measure of the underlying disease and is not confounded by potential symptomatic effects of a study intervention. New developments in understanding the cause of the disease, in the development of animal models of PD, and in clinical trial methodology will hopefully hasten the resolution of these problems. PMID- 19127581 TI - Premotor Parkinson's disease: clinical features, detection, and prospects for treatment. AB - The period immediately before the onset of motor symptoms of Parkinson's disease (PD) often has a recognizable phenotype with features including autonomic dysfunction and impaired olfaction. Subclinical dopaminergic cell loss can also be detected at this time using molecular imaging techniques. A greater recognition of the features of premotor PD and improvements in screening technologies have opened the possibility of predictive testing for PD. In addition to molecular imaging of the dopamine system, screening tests that can potentially be used to identify the physiological abnormalities in premotor PD include olfactory testing, imaging of the sympathetic innervation of the heart, transcranial ultrasound, and genetic testing for mutations known to cause hereditary PD. All of these technologies have trade-offs as screening tests for accuracy, availability, and costs. Using these tests in combination may produce a more favorable combination of reasonable cost and accuracy than using any single test alone. Ultimately, the value of screening for PD depends on development of neuroprotective treatments for PD that would create an imperative for early identification and treatment. PMID- 19127582 TI - Nonmotor manifestations of Parkinson's disease. AB - Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. Traditionally, attention has focused on the motor symptomatology of PD, but it is now appreciated that the nonmotor symptoms affecting neuropsychiatric, sleep, autonomic, and sensory domains occur in up to 88% of PD patients and can be an important source of disability. Nonmotor manifestations of PD play a significant role in the impairment of disease-related quality of life. The cause of nonmotor manifestations of PD is multifactorial, but to a large extent, these manifestations are related to the nature of the neurodegenerative process and the widespread nondopaminergic neuropathological changes associated with the disease. Recognition of nonmotor disability is essential not only for ascertaining the functional status of patients but also for better appreciating the nature of the neurodegenerative process in PD. In addition, a number of nonmotor manifestations can precede the onset of motor symptoms in PD and can be used as screening tools allowing for early disease identification and for trials of possible disease-modifying interventions. This article reviews depression, sleep, and autonomic dysfunction in PD. PMID- 19127584 TI - The basal ganglia in Parkinson's disease: current concepts and unexplained observations. AB - The pathophysiology of Parkinson's disease is reviewed in light of recent advances in the understanding of the functional organization of the basal ganglia (BG). Current emphasis is placed on the parallel interactions between corticostriatal and corticosubthalamic afferents on the one hand, and internal feedback circuits modulating BG output through the globus pallidus pars interna and substantia nigra pars reticulata on the other. In the normal BG network, the globus pallidus pars externa emerges as a main regulatory station of output activity. In the parkinsonian state, dopamine depletion shifts the BG toward inhibiting cortically generated movements by increasing the gain in the globus pallidus pars externa-subthalamic nucleus-globus pallidus pars interna network and reducing activity in "direct" cortico-putaminal-globus pallidus pars interna projections. Standard pharmacological treatments do not mimic the normal physiology of the dopaminergic system and, therefore, fail to restore a functional balance between corticostriatal afferents in the so-called direct and indirect pathways, leading to the development of motor complications. This review emphasizes the concept that the BG can no longer be understood as a "go-through" station in the control of movement, behavior, and emotions. The growing understanding of the complexity of the normal BG and the changes induced by DA depletion should guide the development of more efficacious therapies for Parkinson's disease. PMID- 19127583 TI - Future of cell and gene therapies for Parkinson's disease. AB - The experimental field of restorative neurology continues to advance with implantation of cells or transfer of genes to treat patients with neurological disease. Both strategies have generated a consensus that demonstrates their capacity for structural and molecular brain modification in the adult brain. However, both approaches have yet to successfully address the complexities to make such novel therapeutic modalities work in the clinic. Prior experimental cell transplantation to patients with PD utilized dissected pieces of fetal midbrain tissue, containing mixtures of cells and neuronal types, as donor cells. Stem cell and progenitor cell biology provide new opportunities for selection and development of large batches of specific therapeutic cells. This may allow for cell composition analysis and dosing to optimize the benefit to an individual patient. The biotechnology used for cell and gene therapy for treatment of neurological disease may eventually be as advanced as today's pharmaceutical drug related design processes. Current gene therapy phase 1 safety trials for PD include the delivery of a growth factor (neurturin via the glial cell line derived neurotrophic factor receptor) and a transmitter enzyme (glutamic acid decarboxylase and aromatic acid decarboxylase). Many new insights from cell biological and molecular studies provide opportunities to selectively express or suppress factors relevant to neuroprotection and improved function of neurons involved in PD. Future gene and cell therapies are likely to coexist with classic pharmacological therapies because their use can be tailored to individual patients' underlying disease process and need for neuroprotective or restorative interventions. PMID- 19127585 TI - Functional models of Parkinson's disease: a valuable tool in the development of novel therapies. AB - Functional models of Parkinson's disease (PD) have led to effective treatment for the motor symptoms. Toxin-based models, such as the 6-hydroxydopamine-lesioned rat and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated primate, have resulted in novel dopaminergic therapies and new therapeutic strategies. They have also been used to study processes underlying motor complications, particularly dyskinesia, and for developing pharmacological approaches to dyskinesia avoidance and suppression. Symptomatic models of PD based on nigrostriatal degeneration have a high degree of predictability of clinical effect of dopaminergic drugs on motor symptoms in humans. However, the effects of nondopaminergic drugs in these models do not translate effectively into clinical efficacy. Newer experimental models of PD have attempted to reproduce the pathogenic process and to involve all areas of the brain pathologically affected in humans. In addition, models showing progressive neuronal death have been sought but so far unsuccessfully. Pathogenic modeling has been attempted using a range of toxins, as well as through the use of transgenic models of gene defects in familial PD and mutant rodent strains. However, there are still no accepted progressive models of PD that mimic the processes known to occur during cell death and that result in the motor deficits, pathology, biochemistry, and drug responsiveness as seen in humans. Nevertheless, functional models of PD have led to many advances in treating the motor symptoms of the disorder, and we have been fortunate to have them available. They are an important reason the treatment of PD is so much better compared with treatments for related illnesses. PMID- 19127587 TI - Biomarkers for Parkinson's [corrected] disease: tools to assess Parkinson's disease onset and progression. AB - Reliable and well-validated biomarkers for PD to identify individuals "at risk" before motor symptoms, accurately diagnose individuals at the threshold of clinical PD, and monitor PD progression throughout its course would dramatically accelerate research into both PD cause and therapeutics. Biomarkers offer the potential to provide a window onto disease mechanism, potentially generating therapeutic targets for disease. In particular, biomarkers enable investigation of the premotor period of PD before typical symptoms are manifest, but while degeneration has already begun. Given the multiple genetic causes for PD already identified, the marked variability in the loss of dopaminergic markers measured by imaging at motor symptom onset and the clear heterogeneity of clinical symptoms in PD onset and clinical progression, it is likely many biomarkers with a focus ranging from clinical symptoms to PD pathobiology to molecular genetic mechanisms will be necessary to fully map PD risk and progression. Biomarkers are also critical in new drug development for PD, both in early validation studies to assess drug dosing and to determine drug penetrance into the brain, and in later efficacy studies to complement PD clinical outcomes. During the past two decades, much progress has been made in identifying and assessing PD biomarkers, but as yet, no fully validated biomarker for PD is currently available. Nonetheless, there is increasing evidence that molecular genetics, focused -omic (proteomic, metabolomic, and transcriptomic) assessment of blood and cerebrospinal fluid, and advanced in vivo brain imaging will provide critical clues to assist in the diagnosis and medical management of PD patients. PMID- 19127586 TI - What causes cell death in Parkinson's disease? AB - Currently, there is no proven neuroprotective or neurorestorative therapy for Parkinson's disease (PD). Several advances in the genetics of PD have created an opportunity to develop mechanistic-based therapies that hold particular promise for identifying agents that slow and even halt the progression of PD, as well as restore function. Here we review many of the advances in the last decade regarding the identification of new targets for the treatment of PD based on understanding the molecular mechanisms of how mutations in genes linked to PD cause neurodegeneration. PMID- 19127589 TI - XC1028 from Xanthomonas campestris adopts a PilZ domain-like structure without a c-di-GMP switch. AB - The crystal structure of XC1028 from Xanthomonas campestris has been determined to a resolution of 2.15 A using the multiple anomalous dispersion approach. It bears significant sequence identity and similarity values of 64.10% and 70.09%, respectively, with PA2960, a protein indispensable for type IV pilus-mediated twitching motility, after which the PilZ motif was first named. However, both XC1028 and PA2960 lack detectable c-di-GMP binding capability. Although XC1028 adopts a structure comprising a five-stranded beta-barrel core similar to other canonical PilZ domains with robust c-di-GMP binding ability, considerable differences are observed in the N-terminal motif; XC1028 assumes a compact five stranded beta-barrel without an extra long N-terminal motif, whereas other canonical PilZ domains contain a long N-terminal sequence embedded with an essential "c-di-GMP switch" motif. In addition, a beta-strand (beta1) in the N terminal motif, running in exactly opposite polarity to that of XC1028, is found inserted into the parallel beta3/beta1' strands, forming a completely antiparallel beta4 downward arrow beta3 upward arrow beta1 downward arrow beta1' upward arrow sheet in the canonical PilZ domains. Such dramatic structural differences at the N-terminus may account for the diminished c-di-GMP binding capability of XC1028, and suggest that interactions with additional proteins are necessary to bind c-di-GMP for type IV fimbriae assembly. PMID- 19127590 TI - A generalized knowledge-based discriminatory function for biomolecular interactions. AB - Several novel and established knowledge-based discriminatory function formulations and reference state derivations have been evaluated to identify parameter sets capable of distinguishing native and near-native biomolecular interactions from incorrect ones. We developed the r.m.r function, a novel atomic level radial distribution function with mean reference state that averages over all pairwise atom types from a reduced atom type composition, using experimentally determined intermolecular complexes in the Cambridge Structural Database (CSD) and the Protein Data Bank (PDB) as the information sources. We demonstrate that r.m.r had the best discriminatory accuracy and power for protein small molecule and protein-DNA interactions, regardless of whether the native complex was included or excluded, from the test set. The superior performance of the r.m.r discriminatory function compared with seventeen alternative functions evaluated on publicly available test sets for protein-small molecule and protein DNA interactions indicated that the function was not over optimized through back testing on a single class of biomolecular interactions. The initial success of the reduced composition and superior performance with the CSD as the distribution set over the PDB implies that further improvements and generality of the function are possible by deriving probabilities from subsets of the CSD, using structures that consist of only the atom types to be considered for given biomolecular interactions. The method is available as a web server module at http://protinfo.compbio.washington.edu. PMID- 19127588 TI - Crystal structure of the Fic (Filamentation induced by cAMP) family protein SO4266 (gi|24375750) from Shewanella oneidensis MR-1 at 1.6 A resolution. PMID- 19127591 TI - Comparisons of experimental and computed protein anisotropic temperature factors. AB - Because of its appealing simplicity, the anisotropic network model (ANM) has been widely accepted and applied to study many molecular motion problems: such as ribosome motions, the molecular mechanisms of GroEL-GroES function, allosteric changes in hemoglobin, motor-protein motions, and conformational changes in general. However, the validity of the ANM has not been closely examined. In this work, we use ANM to predict the anisotropic temperature factors of proteins obtained from X-ray and NMR data. The rich, directional anisotropic temperature factor data available for hundreds of proteins in the protein data bank are used as validation data to closely test the ANM model. The significance of this work is that it presents a timely, important evaluation of the model, shows the extent of its accuracy in reproducing experimental anisotropic temperature factors, and suggests ways to improve the model. An improved model will help us better understand the internal dynamics of proteins, which in turn can greatly expand the usefulness of the models, which has already been demonstrated in many applications. PMID- 19127593 TI - Prognostic significance of beta-catenin and topoisomerase IIalpha in de novo acute myeloid leukemia. AB - The Wnt/beta-catenin signaling is important for controlling self-renewal of hematopoietic stem cells and its constitutive activation has recently been documented in a significant proportion of acute myeloid leukemia (AML) cases. Topoisomerase IIalpha (Topo IIalpha) is a marker of cell proliferation and a crucial target for anthracycline cytotoxicity, the mainstay of management employed in AML. We retrospectively investigated the prognostic roles of beta catenin and topo IIalpha in a cohort of 59 patients with newly diagnosed AML by immunohistochemistry. Aberrant beta-catenin expression was demonstrated in 13 patients (22%), and it was more likely to occur in those with unfavorable karyotypes. Advanced age and poor performance status adversely influenced the achievement of complete remission, while neither aberrant beta-catenin expression nor enhanced topo IIalpha activity did. On multivariate survival analysis, four factors independently predicted a shortened overall survival: aberrant beta catenin expression, high topo IIalpha activity, poor-risk cytogenetics, and presence of at least one comorbidity factor. Our results suggest that both beta catenin and topo IIalpha independently predicted an adverse prognosis and might serve as new markers for risk stratification in AML patients. PMID- 19127592 TI - Pseudo-bowel obstruction due to varicella zoster virus infection after autologous stem cell transplantation. PMID- 19127594 TI - Urinary cysteinyl leukotriene E(4) is associated with increased risk for pain and acute chest syndrome in adults with sickle cell disease. AB - Leukotriene E(4) (LTE(4)) levels are associated with rate of pain episodes in children with sickle cell disease (SCD). Because complications of SCD manifest differently in adults than children, we examined a cohort of adults with SCD to determine the relationship between baseline LTE(4) and SCD-related morbidity. Baseline LTE(4) levels were associated with increased rates of pain and acute chest syndrome (ACS) episodes, when those with LTE(4) values in the highest tertile were compared with those in the lowest tertile (pain: risk ratio 7.1, 95% CI 1.8-27.5, P = 0.005; ACS: risk ratio 12.2, 95% CI 2.1-69.8, P = 0.005). PMID- 19127595 TI - Validation of the freezing of gait questionnaire in patients with Parkinson's disease. AB - To revalidate the Freezing of Gait Questionnaire (FOG-Q), patients with Parkinson's disease (PD) were randomly assigned to receive rasagiline (1 mg/day) (n = 150), entacapone (200 mg with each dose of levodopa) (n = 150), or placebo (n = 154). Patients were assessed at baseline and after 10 weeks using the FOG-Q, Unified Parkinson's Disease Rating Scale (UPDRS), Beck Depression Inventory (BDI), and Parkinson's Disease Questionnaire (PDQ-39). FOG-Q dimensionality, test retest reliability, and internal reliability were examined. Convergent and divergent validities were assessed by correlating FOG-Q with UPDRS, BDI, and PDQ 39. Comparisons between FOG-Q item 3 and UPDRS item 14 were also made. Principal component analysis indicated that FOG-Q measures a single dimension. Test-retest reliability and internal reliability of FOG-Q score was high. FOG-Q was best correlated to items of the UPDRS relating to walking, general motor issues, and mobility. Correlations between baseline and endpoint suggested that FOG-Q item 3 is at least as reliable as UPDRS item 14. At baseline, 85.9% of patients were identified as "Freezers" using FOG-Q item 3 (> or =1) and 44.1% using UPDRS item 14 (> or =1) (P < 0.001). FOG-Q was a reliable tool for the assessment of treatment intervention. FOG-Q item 3 was effective as a screening question for the presence of FOG. PMID- 19127596 TI - Rapidly increasing incidence of papillary serous carcinoma of the peritoneum in the United States: fact or artifact? AB - Papillary serous carcinoma of the peritoneum (PSCP) has been recognized for almost 5 decades, but little is known about the etiology or pathogenesis of this uncommon malignancy. The objective of this analysis was to examine trends in the incidence of PSCP in the United States. Invasive PSCP cases (N = 4,389) were identified through 24 population-based registries in the United States during the period 1995-2004. Incidence rates were calculated per million population. PSCP is a disease of older women, with few cases diagnosed before the age of 40 years. The incidence of PSCP was 64% lower among black women and 47% lower among Asian Pacific Islander women compared with white women. Rates among Hispanic women were 39% lower than among non-Hispanic women. The majority of PSCP (68%) was diagnosed at a distant stage, underscoring the difficulty of diagnosing this malignancy. The incidence of PSCP has increased dramatically during the past decade in the United States with the greatest rise (>13% per year) among non-Hispanic and white women. This trend was more pronounced among older women and women with early stage disease. The incidence of PSCP shows substantial racial and ethnic diversity. The increase in the rate of PSCP among all racial and ethnic groups during the 10-year observation period is cause for some alarm. Although the reason for this temporal trend is unknown, some of the increase may be attributable to reclassification of ovarian carcinoma to the peritoneum. PMID- 19127597 TI - MiRNA expression in urothelial carcinomas: important roles of miR-10a, miR-222, miR-125b, miR-7 and miR-452 for tumor stage and metastasis, and frequent homozygous losses of miR-31. AB - We analyzed 34 cases of urothelial carcinomas by miRNA, mRNA and genomic profiling. Unsupervised hierarchical clustering using expression information for 300 miRNAs produced 3 major clusters of tumors corresponding to Ta, T1 and T2-T3 tumors, respectively. A subsequent SAM analysis identified 51 miRNAs that discriminated the 3 pathological subtypes. A score based on the expression levels of the 51 miRNAs, identified muscle invasive tumors with high precision and sensitivity. MiRNAs showing high expression in muscle invasive tumors included miR-222 and miR-125b and in Ta tumors miR-10a. A miRNA signature for FGFR3 mutated cases was also identified with miR-7 as an important member. MiR-31, located in 9p21, was found to be homozygously deleted in 3 cases and miR-452 and miR-452* were shown to be over expressed in node positive tumors. In addition, these latter miRNAs were shown to be excellent prognostic markers for death by disease as outcome. The presented data shows that pathological subtypes of urothelial carcinoma show distinct miRNA gene expression signatures. PMID- 19127599 TI - Morphology controlled growth of chitosan-bound microtubes and a study of their biocompatibility and antibacterial activity. AB - Self-assembled peptide microtubes are fabricated with the biopolymer chitosan. The microtubes are covalently attached to chitosan and the morphology of the chitosan assembled on the surface of the microtubes can be tuned by altering the pH of the growth solution. Cytotoxicity studies in the presence of mouse embryonic fibroblasts indicate that the chitosan-bound microtubes are highly biocompatible and the cells are able to survive and proliferate at a similar rate to the control. Antibacterial studies in the presence of E. coli prove that the chitosan-bound microtubes are bactericidal. This simple method for the development of biocompatible microstructures will facilitate cell targeting, fabrication of efficient carrier devices, and the preparation of highly efficient antibacterial materials. PMID- 19127598 TI - Inhibition of prostaglandin synthesis and actions by genistein in human prostate cancer cells and by soy isoflavones in prostate cancer patients. AB - Soy and its constituent isoflavone genistein inhibit the development and progression of prostate cancer (PCa). Our study in both cultured cells and PCa patients reveals a novel pathway for the actions of genistein, namely the inhibition of the synthesis and biological actions of prostaglandins (PGs), known stimulators of PCa growth. In the cell culture experiments, genistein decreased cyclooxygenase-2 (COX-2) mRNA and protein expression in both human PCa cell lines (LNCaP and PC-3) and primary prostate epithelial cells and increased 15 hydroxyprostaglandin dehydrogenase (15-PGDH) mRNA levels in primary prostate cells. As a result genistein significantly reduced the secretion of PGE(2) by these cells. EP4 and FP PG receptor mRNA were also reduced by genistein, providing an additional mechanism for the suppression of PG biological effects. Further, the growth stimulatory effects of both exogenous PGs and endogenous PGs derived from precursor arachidonic acid were attenuated by genistein. We also performed a pilot randomised double blind clinical study in which placebo or soy isoflavone supplements were given to PCa patients in the neo-adjuvant setting for 2 weeks before prostatectomy. Gene expression changes were measured in the prostatectomy specimens. In PCa patients ingesting isoflavones, we observed significant decreases in prostate COX-2 mRNA and increases in p21 mRNA. There were significant correlations between COX-2 mRNA suppression, p21 mRNA stimulation and serum isoflavone levels. We propose that the inhibition of the PG pathway contributes to the beneficial effect of soy isoflavones in PCa chemoprevention and/or treatment. PMID- 19127600 TI - Preparation of a novel polymer-modified si surface for DNA immobilization. AB - The design of a novel polymer-modified overlayer composed of PPAPE and GPMS on a silicon wafer for immobilization of DNA molecules is described. After hydroxylation of Si(100) surfaces, GPMS molecules were self-assembled onto these surfaces. PPAPE molecules were then covalently attached to the epoxy-terminated surfaces. The incubation time and concentration of PPAPE was found to effect both layer thickness and water CA. The type of organic solvent and the pH were found to change the nature of the PPAPE-modified surface for DNA immobilization. It is concluded that PPAPE-modified surfaces show advantages for DNA immobilization by electrostatic interactions between DNA molecules and positively charged free amino groups of the PPAPE-modified surfaces at the appropriate pH values. PMID- 19127601 TI - Blendable peptide-polymer nanofibers to modulate mechanical properties of polymers. AB - The self-assembly kinetics of a peptide-block-poly(n-butyl acrylate) copolymer (pnBA-(TV){5}{switch}nFGconjugate) in solution and the blend behavior of the resulting fibrillar nanoobjects are investigated. By using a switch peptide segment with temporarily suppressed aggregation behavior, the solution self assembly of the peptide-polymer conjugate can be triggered via a small change in the pH(app.). This results in a controlled assembly of the peptide segments and thus the organization of the bioconjugate. The process was followed by time dependent determination of the dynamic viscosity of the aggregation solution and by AFM that allows visualization of the corresponding structures. The obtained anisotropic nanofibers are characterized in solution and in bulk by SAXS, showing a worm-like characteristic. Blend behavior is investigated, proving the homogeneous dispersibility of the nanofibers in poly(acrylate)s. It could be demonstrated that the nanofibers are modulating the mechanical properties of pnBA by mechanisms of molecular reinforcement, having a distinct stiffening effect. PMID- 19127603 TI - A new concept for more biocompliant bone cements for vertebroplasty and kyphoplasty. AB - A series of more biocompliant bone cements with mechanical properties that match those of different vertebral trabecular or cancellous bones for vertebroplasty and kyphoplasty applications, have been developed. The elastic modulus and strength of the bone cements are in a range of 5 to 400 MPa, and 5 to 75 MPa, respectively. The bone cement shows excellent thermal and dimensional stability in simulated body fluid of Ringer's solution at both 37 and 70 degrees C. The elastic modulus, strength, ultimate strain are constant over time at both temperatures and did not alter dimensions. The bone cement has a much lower exothermic temperature during polymerization compared with commercially available poly(methyl methacrylate) bone cements, and has high cohesiveness in liquid environments. It is able to be delivered through an 18 gauge needle ranging from 4 to 6 inches in length and reaches final mechanical properties within 30 min post delivery. PMID- 19127602 TI - Nickel-ion-mediated control of the stoichiometry of his-tagged protein/nanoparticle interactions. AB - The interaction between synthetic materials and biomolecules plays an important role in biomedical and pathological sciences. An important issue in these interactions is control of stoichiometry. The interaction between NTA ligands and proteins with six consecutive His residues has been widely used for protein purification. Control of stoichiometry is an important issue in applying this recognition strategy to the creation of defined nanoparticle-protein conjugates. In this communication we report the direct control of particle-protein stoichiometry through variation of nickel chloride concentration, as demonstrated through fluorescence and gel electrophoresis. PMID- 19127605 TI - Effects of alignment on interface pressure for transtibial amputee during walking. AB - PURPOSE: To assess the mechanical behaviour at interface for unilateral transtibial amputees during walking when the prosthesis is misaligned, since studies examining interface pressure between residual limb and prosthetic socket have been restricted to unsupported stance and natural gait. METHOD: One male subject with transtibial amputation volunteers for the study. Interface pressures over five sites are measured under three sagittal alignment settings. MP (mean peak interface pressure), TP(90+) (time in which pressure exceeded 90% of peak pressure) and TPI(90+) (time-pressure integral at the period of sustained sub maximal load) are discussed for each alignment setting. RESULTS: Compared with optimal alignment, the trend of interface pressure, the mean peak pressure do not change much,but the duration of sub-maximal pressure changes remarkably, except that at the patellar tendon, and finally the TPI(90+) changes considerably with different alignment settings. CONCLUSIONS: The results offer the clinician and paramedical staff further insight in residual limb/socket interface mechanics in the transtibial amputation patients and provide potentially useful information for socket design and prosthesis fitting. PMID- 19127606 TI - The antenatal identification of funisitis with a rapid MMP-8 bedside test. AB - AIMS: The purpose of this study was to determine if a bedside test, the MMP-8 PTD Check, can be of value in the antenatal identification of funisitis. This test can be performed in 15 min without any laboratory equipment. METHODS: The relationship between the presence or absence of funisitis and the results of an MMP-8 PTD Check was examined in 139 patients who delivered preterm singleton neonates (gestational age <35 weeks) within 72 h of amniocentesis. Amniotic fluid (AF) was cultured for aerobic and anaerobic bacteria and for genital mycoplasmas. AF was analyzed for white blood cell (WBC) count, interleukin-6 (IL-6) and an MMP 8 PTD Check. The IL-6 concentration was also determined in umbilical cord plasma collected at birth. Funisitis was diagnosed in the presence of neutrophil infiltration into the umbilical vessel walls or Wharton's jelly. RESULTS: 1) Funisitis was present in 27% (38/139) of cases; 2) A positive MMP-8 PTD Check had a sensitivity of 97% (37/38), a specificity of 63% (64/101), a positive predictive value of 50% (37/74) and a negative predictive value of 99% (64/65) in the identification of funisitis; 3) Among cases without funisitis, patients with a positive MMP-8 PTD Check had a significantly higher median AF IL-6 concentration, AF WBC count, and umbilical cord plasma IL-6 concentration at birth than those with a negative MMP-8 PTD Check (P<0.05 for each). CONCLUSIONS: The MMP-8 PTD Check is a rapid, simple and sensitive bedside test which allows assessment of the risk of funisitis. PMID- 19127607 TI - Outstanding submissions to the AMIA annual symposium now featured in JAMIA. PMID- 19127608 TI - Though with constraints imposed by endosymbiosis, preferential attachment is still a plausible mechanism responsible for the evolution of the chloroplast metabolic network. AB - Chloroplasts evolved as a result of endosymbiosis, during which sophisticated mechanisms evolved to translocate nucleus-encoded plastid-targeted enzymes into the chloroplast to form the chloroplast metabolic network. Given the constraints and complexity of endosymbiosis, will preferential attachment still be a plausible mechanism for chloroplast metabolic network evolution? We answer this question by analysing the metabolic network properties of the chloroplast and a cyanobacterium, Synechococcus sp. WH8102 (syw). First, we found that enzymes related to more ancient pathways are more connected, and synthetases have the highest connectivity. Most of the enzymes shared by the two densest cores between the chloroplast and syw are synthetases. Second, the highly conserved functional modules mainly consist of highly connected enzymes. Finally, isozymes and enzymes from endosymbiotic gene transfer (EGT) were distributed mainly in conserved modules and showed higher connectivity than nonisozymes or non-EGT enzymes. These results suggest that even with severe evolutionary constraints imposed by endosymbiosis, preferential attachment is still a plausible mechanism responsible for the evolution of the chloroplast metabolic network. However, the current analysis may not completely differentiate whether the chloroplast network properties reflect the evolution of the chloroplast network through preferential attachment or has been inherited from its cyanobacterial ancestor. To fully differentiate these two possibilities, further analyses of the metabolic network structure properties of organisms at various intermediate evolutionary stages between cyanobacteria and the chloroplast are needed. PMID- 19127609 TI - Human-induced changes in the reproductive traits of Lake Constance common whitefish (Coregonus lavaretus). AB - Size-selective fishery harvest leads to phenotypic changes in fish reproductive traits. When these changes represent an evolutionary response of a stock, they may have severe consequences for future stock dynamics and yields. In freshwater ecosystems, reproductive traits may also be affected by other human impacts such as changes in system productivity. The present study uses regression analysis to evaluate the impacts of changes in lake trophy and of an intensive size-selective harvest over several decades on the reproductive traits of common whitefish in Lake Constance between 1963 and 1999. Fecundity was strongly linked to lake trophy but was also related to the calendar year, suggesting an evolutionary response to size-selective harvest and to massive stocking of the lake with hatchery-reared larvae. The present study is an example of how fish reproductive traits are influenced by the combined action of various human impacts: changes in system productivity, size-selective harvest and massive stocking. PMID- 19127610 TI - Five questions on ecological speciation addressed with individual-based simulations. AB - We use an individual-based simulation model to investigate factors influencing progress toward ecological speciation. We find that environmental differences can quickly lead to the evolution of substantial reproductive barriers between a population colonizing a new environment and the ancestral population in the old environment. Natural selection against immigrants and hybrids was a major contributor to this isolation, but the evolution of sexual preference was also important. Increasing dispersal had both positive and negative effects on population size in the new environment and had positive effects on natural selection against immigrants and hybrids. Genetic divergence at unlinked, neutral genetic markers was low, except when environmental differences were large and sexual preference was present. Our results highlight the importance of divergent selection and adaptive divergence for ecological speciation. At the same time, they reveal several interesting nonlinearities in interactions between environmental differences, sexual preference, dispersal and population size. PMID- 19127611 TI - Fitness and the level of homozygosity in a social insect. AB - To date very few studies have addressed the effects of inbreeding in social Hymenoptera, perhaps because the costs of inbreeding are generally considered marginal owing to male haploidy whereby recessive deleterious alleles are strongly exposed to selection in males. Here, we present one of the first studies on the effects of queen and worker homozygosity on colony performance. In a wild population of the ant Formica exsecta, the relative investment of single-queen colonies in sexual production decreased with increased worker homozygosity. This may either stem from increased homozygosity decreasing the likelihood of diploid brood to develop into queens or a lower efficiency of more homozygous workers at feeding larvae and thus a lower proportion of the female brood developing into queens. There was also a significant negative association between colony age and the level of queen but not worker homozygosity. This association may stem from inbreeding affecting queen lifespan and/or their fecundity, and thus colony survival. However, there was no association between queen homozygosity and colony size, suggesting that inbreeding affects colony survival as a result of inbred queens having a shorter lifespan rather than a lower fecundity. Finally, there was no significant association between either worker or queen homozygosity and the probability of successful colony founding, colony size and colony productivity, the three other traits studied. Overall, these results indicate that inbreeding depression may have important effects on colony fitness by affecting both the parental (queen) and offspring (worker)generations cohabiting within an ant colony. PMID- 19127612 TI - Lovesick: immunological costs of mating to male sagebrush crickets. AB - A growing body of evidence suggests that resources invested in reproduction often come at the expense of the ability to mount an immune response. During mating, female sagebrush crickets, Cyphoderris strepitans, consume the ends of the male's hind wings and ingest his haemolymph. Previous research has shown that this behaviour impairs the ability of males to secure additional matings. One hypothesis to account for this effect is that wing wounding triggers an energetically costly immune response, such that nonvirgin males are unable to sustain the costly acoustical signalling needed to attract additional females. To test this hypothesis, we injected virgin males with lipopolysaccharides (LPS) to provoke an immune response, and monitored their mating success in the field. LPS injected virgin males took significantly longer to mate than sham-injected virgin males, and spent significantly less time calling. We also compared virgin, nonvirgin and experimentally wing-wounded virgin males with respect to: (1) their ability to encapsulate a foreign invader via the accumulation of haemocytes and deposition of melanin and (2) baseline levels of phenoloxidase (PO), a key enzyme in the biochemical cascade leading to the production of melanin. Although encapsulation ability did not differ with reproductive experience, virgin males had significantly higher levels of PO than either nonvirgin or experimentally wing-wounded virgin males. These results suggest that wing-wounding alone is sufficient to impair male immunity, and that males trade-off investment in reproduction and immunity. PMID- 19127613 TI - The Diabetic Foot in 2008: an update from the 12th Malvern Diabetic Foot Meeting. AB - Every 2 years, Great Malvern, a small town in the county of Worcestershire, England, is home to the oldest international meeting on the diabetic foot that started about a quarter of a century ago. The 12th Malvern Diabetic Foot meeting, held from May 14 to 16, 2008, was attended by delegates and speakers from 5 continents and more than 25 countries. Strategic clinical specialties involved in diabetic foot management were prominently represented. Virtually every aspect of diabetic foot care was covered during this 3-day meeting through invited plenary lectures, practical interactive workshops, and scientific oral presentations. The evidence base underpinning current foot care was critically appraised, and topical issues on the diabetic foot were revisited. This review puts together key messages emanating from the meeting. All effort has been made to highlight clearly where consensus exists as well as outline areas where controversy persists. PMID- 19127614 TI - [Prestigious names and significant contributions in MMW. A world wide medical journal]. PMID- 19127615 TI - [Transition, revision, professionalization]. PMID- 19127616 TI - [Fusion, family practitioner information, new image text]. PMID- 19127617 TI - [Cosmetic revision of the face of MMW]. PMID- 19127618 TI - [125 years "Progress in Medicine"]. PMID- 19127619 TI - [Retrospect and prospects of 20 years EMB reforms. What have they brought for the contract physician?]. PMID- 19127620 TI - [The future of continuing medical education. CME credits as world wide standard]. PMID- 19127621 TI - [Physician adjustment to change]. PMID- 19127622 TI - [Primary care at risk. Quo vadis family physician?]. PMID- 19127623 TI - ["As fast as work on a conveyer belt". The exploitation of the physician resource]. PMID- 19127624 TI - [Fine filter for information deluge]. PMID- 19127625 TI - [25 years of progress in pulmonology: inhalation therapy defeats dyspnoea]. PMID- 19127626 TI - [25 years of progress in gastroenterology: the identification of pathogens led to curative strategies]. PMID- 19127627 TI - [25 years of progress in gastroenterology--diseases of the bowel: colonoscopy gives tumour prevention a chance]. PMID- 19127628 TI - [25 years of progress in endocrinology: treating with drugs instead of mutilating the pituitary gland]. PMID- 19127629 TI - [25 years of progress in geriatric medicine: how much therapy can be tolerated by the patient?]. PMID- 19127630 TI - [25 years of progress in rheumatology: quantum jumps in the treatment of rheumatoid arthritis]. PMID- 19127631 TI - [25 years of progress in infectiology: new approaches in the fight against resistant pathogens]. PMID- 19127632 TI - [25 years of progress in natural medicine: even a gentle discipline requires hard data]. PMID- 19127633 TI - Transitional cell carcinoma of the kidney invading the inferior vena cava, treated by excision and grafting, with a review of vena cava replacement. PMID- 19127635 TI - When to see a doctor. PMID- 19127634 TI - Reconstruction of penile shaft amputation: is microvascular re-anastomosis mandatory? PMID- 19127636 TI - Two prescriptions for America's ills: McCain and Obama offer conflicting health plans. Here's how you'd fare. PMID- 19127637 TI - Pyrexia of unknown origin associated with bladder carcinoma. PMID- 19127638 TI - Non-metastatic primitive peripheral neuroectodermal tumour of the kidney (extraskeletal Ewing's sarcoma) with vena caval tumour thrombus. PMID- 19127639 TI - Finasteride in polycythaemia. PMID- 19127640 TI - Metastatic adenocarcinoma of the prostate with a testicular mass and cutaneous nodules. PMID- 19127641 TI - Pyoderma gangrenosum of the penis: an interesting penile lesion. PMID- 19127642 TI - 'Egg-shell' encrustation over a Foley balloon: an unusual cause of urinary retention after urethroplasty. PMID- 19127643 TI - Coexistence of urachal and dorsal urethral anomalies: a rare occurrence. PMID- 19127644 TI - Collateral urethral duplication in the frontal plane. PMID- 19127645 TI - Persistent urogenital sinus with bladder agenesis and absence of vagina. PMID- 19127646 TI - Double-dose intravenous pyelogram voiding urography to delineate stricture disease. PMID- 19127647 TI - Let the patient decide. PMID- 19127648 TI - Primary adenoid cystic carcinoma arising in the region of Cowper's gland. PMID- 19127649 TI - Calorimetric evaluations of Bacillus subtilis vegetative and spore cells colonial growth and suppressive effects of sucrose monopalmitate. AB - The colonial growth of Bacillus subtilis vegetative and spore cells was evaluated by determining the time course of the growth heat changes (growth thermogram) with a microbial calorimeter. The actual heat evolution curve (f (t) curve) obtained from the thermogram was in good agreement with the viable cell number changes. From the logarithmic f (t) curve, the exponential growth phase was estimated to be the period of between 31 and 35 h after inoculation and the growth rate constant (mu') was 0.36 h(-1). The f (t) curve was also obtained from the thermogram of the spore cells. The starting time of growth of the spore cells was 9 h later than that of the vegetative cells. This period corresponded to the germination period. The growth profile after germination was almost similar to that of the vegetative cells. The suppressive activities of sucrose monopalmitate (SMP) against the vegetative and spore cells were evaluated by the f (t) curves. Our results show that SMP inhibited metabolic inductions during the lag phase of the vegetative cells and the germination of the spore cells. Microbial calorimetry is a method suitable for non-destructive growth measurements of microbial colonies on the surface of a solid medium. PMID- 19127650 TI - Development of a small-sized generator of ozonated water using an electro conductive diamond electrode. AB - A small-sized generator of ozonated water was developed using an electro conductive diamond. We studied the optimum conditions for producing ozonated water. As a result, we developed a small-sized generator of ozonated water driven by a dry-cell for use in the average household. This generator was easily able to produce ozonated water with an ozone concentration (over 4 mg/L) sufficient for disinfection. In addition, we verified the high disinfecting performance of the water produced in an actual hospital. PMID- 19127651 TI - RAPD analysis of salt-tolerant yeasts from contaminated seasoned pickled plums and their growth inhibition using food additives. AB - Eight salt-tolerant yeasts were isolated from contaminated pickled plums which were seasoned with honey and "Umami" seasoning. They were classified into four main groups according to random amplified polymorphic DNA analysis, and three of ten kinds of food additives tested inhibited their growth. The type strains of each group were identified as Zygosaccharomyces bisporus, Pichia subpeliculosa, and two strains of Candida apicola based on the D1/D2 region sequence of the 26S rRNA gene. They were able to grow in medium containing 6% (w/v) NaCI. A number of yeasts were isolated from production lines by the swab method, but not from the salted plums used as raw materials. These results show that the production lines require washing with antimicrobial agents effective against salt-tolerant yeasts. Three commercial food additives, San-keeper 381, Sunsoft No.700P-2, and potassium sorbate inhibited the growth of Z. bisporus at 125 to 250 microg/ml. In particular, San-keeper 381 altered the morphology of this species at 125 microg/ml. C. apicola and P. subpelliculosa were inhibited by Sunsoft No.700P-2 and potassium sorbate at 250 microg/ml. These results indicate that the washing of production lines with disinfectant and the use of food additives that effectively prevent salt-tolerant yeast contamination are necessary. PMID- 19127652 TI - Antiviral activities of heated dolomite powder. AB - The effect of the heating conditions of dolomite powder on its antiviral activity was studied against the H5N3 avian influenza virus. Calcium oxide (CaO) and magnesium oxide (MgO), obtained by the thermal decomposition of dolomite above 800 degrees C, were shown to have strong antiviral activity, but the effect was lessened when the heating temperature exceeded 1400 degrees C. Simultaneous measurement of the crystallite size suggested that the weakening of the activity was due to the considerable grain growth of the oxides. It was found that the presence of Mg in dolomite contributed to the deterrence of grain growth of the oxides during the heating process. Although both CaO and MgO exhibited strong antiviral activity, CaO had the stronger activity but quickly hydrated in the presence of water. On the other hand, the hydration of MgO took place gradually under the same conditions. Separate measurements using MgO and Mg(OH)2 revealed that MgO had a higher antiviral effect than Mg(OH)2. From the overall experiments, it was suggested that the strong antiviral activity of dolomite was related to the hydration reaction of CaO. PMID- 19127653 TI - Survival of pathogenic Yersinia enterocolitica in vacuum-packed or non-vacuum packed pork at low temperature. AB - Pathogenic Yersinia enterocolitica serotypes O:3, O:5,27, O:8 and O:9 were inoculated into sliced and ground pork, and the samples were stored under vacuum or aerobic conditions at 2 and degrees C. All serotypes survived for 5 weeks in pork with or without vacuum packing without any discernable increases in their population. In sterilized pork with or without vacuum packing, there was no evident growth of Y. enterocolitica. In pork broth in which the pH had been artificially adjusted to 6.8, the growth of Y. enterocolitica was faster than that at 5.7. It is suggested that the growth of Y. enterocolitica in pork with or without vacuum packing may be inhibited by pH but not by the microflora or lactic acid bacteria in pork. PMID- 19127654 TI - Autism profiles of males with fragile X syndrome. AB - Autism, which is common in individuals with fragile X syndrome, is often difficult to diagnose. We compared the diagnostic classifications of two measures for autism diagnosis, the ADOS and the ADI-R, in addition to the DSM-IV-TR in 63 males with this syndrome. Overall, 30% of the subjects met criteria for autistic disorder and 30% met criteria for PDD-NOS. The classifications on the ADOS and DSM-IV-TR were most similar, whereas the ADI-R classified subjects as autistic much more frequently. We further investigated the relationship of both FMRP and FMRI mRNA to symptoms of autism in this cohort and found no significant relationship between the measures of autism and molecular features, including FMRP, FMRI mRNA, and CGG repeat number. PMID- 19127655 TI - Evidence for latent classes of IQ in young children with autism spectrum disorder. AB - Autism is currently viewed as a spectrum condition that includes strikingly different severity levels; IQ is consistently described as one of the primary aspects of the heterogeneity in autism. To investigate the possibility of more than one distinct subtype of autism based on IQ both latent class analysis and taxometrics methods were used to classify Mullen IQs in a sample of 456 children with autism spectrum disorder. We found evidence for multiple IQbased subgroups using both methods. Groups differed in level of intellectual functioning and patterns of verbal versus nonverbal ability. Results support the notion of distinct subtypes of autism that differ in severity of intellectual ability, patterns of cognitive strengths and weaknesses, and severity of autism symptoms. PMID- 19127656 TI - Memory skills of boys with fragile X syndrome. AB - Multiple aspects of memory were examined in 42 boys with fragile X syndrome and a comparison group of 42 typically developing boys matched on MA. Working memory, incidental memory, and deliberate memory were assessed with a battery that included both free-recall and recognition tasks. Findings indicated that boys with fragile X syndrome performed more poorly than their matches on most measures. The exception was free recall, in which their accuracy was equal to that of the control participants. Results from analyses of a subset of boys with fragile X syndrome who exhibit characteristics of autism and their MA matches, though preliminary, support the conclusion that memory deficits are especially marked in boys who have fragile X syndrome and evidence autistic behaviors. PMID- 19127657 TI - Gatekeepers of science: attitudes toward the research participation of adults with intellectual disability. AB - Researchers and Institutional Review Board (IRB) members' attitudes influence scientific knowledge about individuals with intellectual disability. We recruited 260 intellectual disability researchers and IRB members to develop a measure of attitudes toward the research participation of adults with intellectual disability, the Participation in Research Attitude Scale. Findings suggest three conceptual domains: Opportunity and Choice, Help in Decision Making, and Beneficence. We also examined individual differences in attitudes and the relationships between general and specific attitudes. In general, intellectual disability researchers and those with closer relationships to individuals with disabilities had attitudes consistent with disability-rights principles. Some dimensions of global attitudes toward adults with intellectual disability predictedmore specific attitudes toward their research participation. Implications are discussed. PMID- 19127658 TI - Peer interactions and academic engagement of youth with developmental disabilities in inclusive middle and high school classrooms. AB - We examined the peer interactions and academic engagement of 23 middle and high school students with developmental disabilities within inclusive academic and elective classrooms. The extent to which students with and without disabilities interacted socially was highly variable and influenced by instructional format, the proximity of general and special educators, and curricular area. Peer interactions occurred more often within small group instructional formats, when students were not receiving direct support from a paraprofessional or special educator, and in elective courses. Academic engagement also varied, with higher levels evidenced during one-to-one or small group instruction and when in proximity of general or special educators. Implications for designing effective support strategies for students with autism and/or intellectual disability within general education classrooms are discussed. PMID- 19127659 TI - Strategic approach to collecting the patient's portion. PMID- 19127660 TI - Using technology to improve patient collections. PMID- 19127661 TI - Mission: reduce employee turnover; game plan: employee education. PMID- 19127662 TI - Indian Society for Malaria and Other Communicable Diseases "Honorary Fellowship" address by Dr Samlee Plianbangchang, Regional Director, World Health Organizaion, South East Asia Region. PMID- 19127663 TI - Evaluation of the WHO clinical case definition of AIDS among children in India. AB - The need of a clinical case definition (CCD) for Acquired Immunodeficiency Syndrome (AIDS) was felt by public health agencies to monitor diseases resulting from human immunodeficiency virus (HIV) infection. To test the statistical significance of the existing World Health Organization (WHO) CCD for the diagnosis of AIDS in areas where diagnostic resources are limited in India, a prospective study was conducted in the Paediatrics department at Dr. Ram Manohar Lohia Hospital, New Delhi. 360 cases between 18 months-12 years of age satisfying WHO case definitions of AIDS were included in the study group. Informed consent was taken from the parents. The serum of patients was subjected to ELISA to conform the diagnosis of HIV infection. Our study detected 16.66% (60) of HIV prevalence in children visiting paediatrics outpatient clinic. 20% cases manifested 3 major and 2 minor signs. This definition had a sensitivity of 73.33%, specificity of 90.66%, positive predictive value (PPV) of 61.11% and negative predictive value (NPV) of 94.44%. On using stepwise logistic regression analysis weight loss, chronic fever > 1 month and total lymphocyte count of less than 1500 cells/mm3 emerged as important predictors. Cases showing 2 major and 2 minor signs were 86 (23.89%) with a sensitivity and specificity of 86.66% and 88.66% respectively. Based on these findings, we propose a clinical case definition based on 13 clinical signs and symptoms for paediatric AIDS in India with better sensitivity and PPV than the WHO case definition but with almost similar specificity. Thus multicentric studies are further required to modify these criteria in Indian set up. PMID- 19127664 TI - Locomotor disability in bancroftian filarial lymphoedema patients. AB - Disability prevention in cases with lymphatic filarial disease has been a research priority in view of the current programme on global elimination of lymphatic filariasis. This is also important for estimation of disease burden, impact evaluation, developing strategy for morbidity management and rehabilitation for lymphoedema patients in the endemic countries. In this communication, we are presenting the results of quantification of functional limitations of lower extremities with lymphoedema caused due to bancroftian filariasis by objective assessment of movement of joints and power of muscles in the affected legs. A total of 81 consecutive lymphoedema patients attending a filariasis clinic in Pondicherry urban locality were recruited for the study. Assessment for restriction of movement of joints and loss of power of muscles was carried out as in Manual for doctors to evaluate permanent physical impairment, prescribed by an expert group of WHO and Ministry of Health, Govt. of India on evaluation for permanent disability. Of the cases assessed, 40% in grade-I, 55% in grade-II, 77.3% in grade-III and 94.7% in grade-IV lymphoedema cases had functional limitations either in joint movements or power of muscles or both. The effective loss of locomotor/function (combined loss of joint movement and power of muscles in %) increased with stage of lymphoedema (grade-I-4.3+7.4, grade-II 7.0+8.4, grade-III-15.4+14.8 and grade IV- 33.2+22.8). The degree of loss varied significantly between the grades (P< 0.0001). The methodology used in this study can be adapted to evaluate the impact of the morbidity management component of strategy for Elimination of Lymphatic Flariasis (ELF) programme. This study will also enable researchers for fine-tuning the method for estimating disease burden and, to develop and evaluate strategies for morbidity management/rehabilitation of filarial lymphoedema patients. PMID- 19127665 TI - Occurrence and detection of AmpC beta lactamases among clinical isolates of E. coli and K. pneumoniae causing UTI. AB - Presence of Bush class C enzymes in uropathogenic strains of Klebsiella pneumoniae & E. coli resistant to extended spectrum cephalosporins is an emerging threat to clinical therapeutics. These resistant strains result in considerable treatment failure and cannot be detected by routine antibiotic sensitivity screening methods. An effort was therefore made to study AmpC beta lactamase production in E. coli and Klebsiella pneumoniae strains causing UTI. A total of 126 E. coli and 49 K. pneumnoniae strains isolated from urine samples were selected for study out of which AmpC beta lactamase production was seen in 23% E. coli (29 isolates) and 18% K. pneumoniae (49 isolates). The susceptibility of AmpC beta lactamase producers to Imipenem, Nitrofurantoin and Amikacin was found to be 100%, 92% and 80% respectively. Thereby the present study emphasizes the importance of monitoring and control of usage of extended spectrum cephalosporins. PMID- 19127666 TI - Epidemiology of animal bites and rabies cases in India. A multicentric study. AB - Rabies, a disease of antiquity continues to be a major public health problem in India. Multiple factors contribute to high mortality and morbidity due to animal bites. An effective strategy for control of rabies takes into account the epidemiology of animal bites, rabies and factors influencing post exposure treatment. The study was carried out as a part of Agreement for Performance of Work (APW) from World Health Organization (WHO) during the period April 2001 to September 2002. Two sets of proformae were developed and used after field testing to interview cases of animal bites and get retrospective information about rabies cases. The study was carried out at six selected centres across the country viz. Delhi, Hyderabad, Raipur, Jamnagar, Coonoor and Rajahmundry and was co-ordinated by National Institute of Communicable Diseases (NICD), Delhi. The officials engaged in the study work were thoroughly trained in the study methodology before the start of the study itself. To maintain quality and uniformity supervisory checks were done during the survey. A total of 1357 fresh animal bite victims were interviewed (exit interview) from the anti-rabies centres (ARCs). Dog bites caused maximum morbidity (92%). Second most common biting animal was monkey (3.2%), followed by cat (1.8%), fox (0.4%) etc. Most bites (64.3%) were unprovoked bites by stray (64.7%) animals. In this study 72.4% animal bite victims were males and 47.5% were children in age group of 2-18 years. 63% had Category III exposure as per the WHO classification. Before coming to ARCs 58.5% people had washed the wound with water/soap or water alone. Some of the bite victims (10.8%) had also applied chillies, salt, turmeric powder, lime, snuff powder, paste of leaves, acid, ash given by Peer Baba (magician) etc. These practices varied from one region to another. The practice of wound washing at the ARC which is an important component of animal bite management was being practiced at only one of the six centres. Of the six centres, Rabies Immunoglobulin (RIG) was available and was being used at only two centres. The study was conducted in public sector ARCs where Nervous Tissue Vaccine (NTV) was available free of cost. All the centres were using NTV except Coonoor, which is using indigenously produced Tissue Culture Vaccine along with NTV. Analysis of 192 case records of rabies cases, from two centres, revealed that dog bites caused maximum mortality (96.9%). Nearly 40% were children below 15 years of age and 78.6% were males indicating that it is an exposure related disease. In all cases, failure to seek timely and appropriate treatment led to development of disease. This paper provides an overview of epidemiology of animal bites and retrospective information about rabies patients. There is a need to strengthen Information, Education and Communication (IEC) programme regarding merits of local wound management including "do's and don'ts". ARCs should be strengthened in terms of facilities and availability of safe and effective anti rabies immunobiologicals. There is a need to create awareness regarding epidemiology and at-home and hospital management of animal bites among the service providers and general community. PMID- 19127667 TI - Awareness of health personnel about lymphatic filariasis and mass drug administration in Kerala State. AB - The mass drug administration programme to eliminate lymphatic filariasis with DEC in Kerala was started in 1997, extended to all the 11 endemic districts by 2005. Since the beginning of Mass drug Administration, the drug consumption rate was found to be not satisfactory. The reasons for noncompliance indicated that the community is not fully convinced about the programme. The knowledge of the medical and para medical workers is certainly a factor in the success of implementation of the programme and is vital. To ascertain the knowledge, a study was undertaken and found not satisfactory. Hence intensive training on all aspects of lymphatic filariasis and the Mass drug Administration programme to achieve the requisite drug consumption rate to meet the goal is needed. PMID- 19127668 TI - Present susceptibility status of rat flea Xenopsylla cheopis (Siphonaptera: Pulicidae), vector of plague against organochlorine, organophosphate and synthetic pyrethroids 1. The Nilgiris District, Tamil Nadu, India. AB - The susceptibility status of Xenopsylla cheopis, the efficient vector of human plague in India was assessed in erstwhile plague endemic areas of Nilgiris district, Tamil Nadu following standard WHO techniques. The studies revealed the development of resistance in rat fleas to DDT--4.0%, Malathion--5.0%, Deltamethrin--0.05% and tolerance to Permethrin--0.75% in all the four blocks of Nilgiris hill district. Development of resistance may be due to the extensive use of insecticides in tea plantations and agricultural sectors where the domestic/peri-domestic rodents find their natural habitats and intermingle with each other. PMID- 19127669 TI - Auramine phenol staining of smears for screening acid fast bacilli in clinical specimens. AB - Tuberculosis is a major threat to the public health allover world. Out of the total tuberculosis cases reported globally, more than half are reported from African continent and India. Two to three fold rises in tuberculosis cases has been reported in the last two decades. Early diagnosis and treatment is one of the effective tools to control the rapid spread of disease. The aim of this study was to find out the value of auramine phenol (AP) staining technique in diagnosis of the suspected tuberculosis cases. A total of 2000 samples which included sputum (746), gastric aspirates (380), urine (336), endometrial biopsy (150), pleural fulids (146), Synovial fluids (67), ascitic fluids (35), cerebrospinal fluids (43), bone marrow (18), lymph node biopsy (11), pericardial aspirates (6), skin biopsy (4), peritoneal fluids (2), and stool (1) were included in the study. Sample were subjected for decontamination procedure by using standard Petroffs method. The deposit smears were stained by auramine phenol (AP stain) and Ziehl Neelsenstaining (ZN stain) and specimens were cultured for Mycobacterium tuberculosis. Of the total positive isolates 69.23% were having pulmonary tuberculosis and 30.76 had extrapulmonary tuberculosis Genitourinary tuberculosis was the most common diagnosis among the extrapulmonary tuberculosis followed by chronic synovitis, bursitis, meningitis, septic arthritis and pericardial effusion. Out of 130 positive samples 70 by culture, 66 smears were positive by auramine phenol stain and 62 were positive by ZN stain. A total of 27 samples were tested positive only by AP staining technique, which included (12) pulmonary and (15) extrpulmonary samples. The endometrial biopsy and pericardial fluid samples showed positive for acid fast bacilli by AP stain only, whereas ZN stain and culture technique failed to demonstrate any bacilli in the same sample. Auramine stain showed high sensitivity (47.14%) and specificity (96.58%). Result of the present study showed that the auramine stain is a better method for screening samples from the suspected cases of tuberculosis sample especially pulmonary and extrapulmonary cases where bacilli count is usually low. PMID- 19127670 TI - A study on determinants of occurrence of complications and fatality among diphtheria cases admitted to ID & BG Hospital of Kolkata. AB - This was a hospital-based case series study of diphtheria patients admitted to ID & BG Hospital of Kolkata from May 2004 to April 2005. A total of 107 patients were included in the study. Report of follow up of admitted patients and interview of family members were done. The study results showed that the highest number of cases (31) were among the 6-10 year age group, while the lowest was among the 16-20 year age group (12). Overall case fatality was high (12.1%). Lowest case fatality (5%) among 21 years and above age group. Case fatality was less among those patients who were admitted to the hospital within 48 hours of occurrence of clinical features. Majority of the patients (72) developed complications. Among them, only Myocarditis occurred in 51, Myocarditis with Palatal palsy in 11, only Palatal palsy in 7, while Respiratory distress occurred in 3. Most of the admitted patients were non-immunized (84). Majority of the respondents (73) have no idea regarding the mode of spread of the disease. Only one respondent had correct knowledge about the benefit of the DPT vaccine. The main reason for non-immunization was ignorance (44%). IEC for full immunization coverage and early admission with proper management are indispensable for the reduction of diphtheria morbidity and mortality. PMID- 19127671 TI - An epidemiological study on incidence, symptoms and complications of measles in bordering districts of West Bengal. AB - The present study was carried out to find out the incidence of measles in under five children and its complications in four selected urban slums of inter-state border districts of West Bengal. 20 cluster sampling technique was followed and in each cluster 250 under five children were covered. Paramedical workers found out the children of the target age group who had the history of measles in past one year and Medical Officer confirmed the case following standard case definition. Incidence of measles was round to be 3.3% in Purulia, 5.5% in Bankura. 4.6% in Midnapur, 5.7% in Haldia-Tamluk and with an overall rate of 4.8%. Incidence was higher in 0-11 and 12-23 months age group and decreased with increasing age, but no sex difference in incidence of Measles was observed. Only 20% of cases were vaccinated with measles vaccine and 26% received Vitamin A in oil. Diarrhoea was the commonest complications followed by cough and cold, pneumonia and weight loss. The study highlighted the necessity of timely measles vaccine coverage, additional dose at a higher age and Vitamin A oil supplementation through IEC activities. PMID- 19127672 TI - Enteric protozoan parasitic infection in HIV seropositive individuals with diarrhoea. AB - A total of 78 HIV seropositive patients were included in the study from January 2004 to March 2006. Enteric protozoan parasitic infections were diagnosed by microscopic examination of stool by fresh wet mount, stool smears stained by modified Z.N., Trichrome and Calcofluor white stains. 20 samples (25.6%) revealed presence ofenteric protozoan parasites. PMID- 19127673 TI - Knowledge and awareness about hepatitis B infection in newly admitted medical students. PMID- 19127674 TI - Awareness of management of tuberculosis among health workers in a rural area of West Bengal. PMID- 19127675 TI - Tuberculosis of the ilium: a case report. AB - A twenty year old man presented with pain in the upper part of right hip. CT scan showed multiple osteolytic areas in the right ilium, suggesting malignancy. CT guided needle biopsy was done and diagnosis of tuberculosis was made. The patient was cured with medical management only. Isolated tuberculosis of the ilium is an extremely rare condition. It is important to have a high clinical suspicion to diagnose early and manage the patient without surgical intervention. PMID- 19127676 TI - Indian tick typhus mimicking as Rocky Mountain Spotted Fever: a case report. PMID- 19127677 TI - Two cases of Kala-azar in Haryana with no evidence of local transmission. PMID- 19127678 TI - Interpersonal relationship. PMID- 19127679 TI - Nurses in politics. PMID- 19127680 TI - Women's knowledge on reproductive tract infections in selected area of Raichur. PMID- 19127681 TI - Primary extraskeletal osteosarcoma of the glans penis and glanular reconstruction. PMID- 19127682 TI - Rx for BlackBerry thumb. PMID- 19127683 TI - Masturbation inducing synchronous bilateral testicular torsion in an adolescent. PMID- 19127684 TI - Renal sinus haemangioma with inferior vena cava involvement. PMID- 19127685 TI - Shake salt from your diet: but beware the stuff hides in strange places. PMID- 19127686 TI - Effective combined chemotherapy for prostatic ductal adenocarcinoma: continuous venous infusion of 5-fluorouracil and low-dose consecutive cisplatin. PMID- 19127687 TI - Adenoid cystic carcinoma of Cowper's gland. PMID- 19127688 TI - Intravascular papillary endothelial hyperplasia in a female urethral mass. PMID- 19127689 TI - Deep haemangioma of the scrotum as a rare presentation of scrotal mass in children. PMID- 19127690 TI - [Epidemiological transition, urban infrastructures, and development: the city of Porto]. PMID- 19127691 TI - [Influencing the prescriptions of general practitioners via guidelines and feedback--secondary publication]. AB - We investigated the effects of a multi-dimensional intervention on practice prescription patterns in five drug groups. The number of prescribed defined daily doses (DDDs) increased after the intervention, while potential savings/DDD decreased. The county's average cost/DDD fell to a level below the national average. It is possible to change general practitioners' prescription patterns without interfering with their clinical freedom or patients' access to treatment. PMID- 19127692 TI - Change in stature of North Indians from British Rule to early independence. PMID- 19127693 TI - [The body of the "socialist woman": models of womanhood in the Soviet occupation zone and the early years of East Germany]. PMID- 19127694 TI - [Endovascular treatment]. PMID- 19127695 TI - ["Who does watch the head watcher"--who can and will watch the experts treatment?]. PMID- 19127696 TI - [Use of media by physicians to criticize their colleagues' work]. PMID- 19127697 TI - [The chances help]. PMID- 19127698 TI - Psychology and schooling: the impact of Susan Isaacs and Jean Piaget on 1960s science education reform. PMID- 19127699 TI - [Psychotic--to be or not to be]. PMID- 19127701 TI - [Psychodynamic psychotherapy in psychotic disorders]. PMID- 19127700 TI - "A strange mixture of caring and corruption": residential care in Christian Brothers orphanages and industrial schools during their last phase, 1940s to 1960s. PMID- 19127702 TI - [Professionalism and quality in the treatment of eating disorders]. PMID- 19127703 TI - Why did South Wales miners have high mortality? Evidence from the mid-twentieth century. PMID- 19127704 TI - The limits of persuasion: advertising, gender and the culture of Australian smoking. AB - This article examines the shift from pipe to cigarette smoking in Australia from World War I to the 1950s; challenges the assumption that women were the major source of the rising popularity of cigarette smoking; questions, through a content analysis of themes in cigarette advertisements in major newspapers, the impact on smoking patterns of mass advertising before the 1950s; and shows the rising importance of roll-your-own cigarette consumption as a sign of masculine and working-class identity and as an adaptation to modern urban life. PMID- 19127705 TI - F. L. McDougall and the origins of the FAO. PMID- 19127706 TI - Inflammatory and oxidative and nitrosative stress pathways underpinning chronic fatigue, somatization and psychosomatic symptoms. AB - PURPOSE OF REVIEW: The aim of this paper is to review recent findings on inflammatory and oxidative and nitrosative stress (IO&NS) pathways in chronic fatigue and somatization disorder. RECENT FINDINGS: Activation of IO&NS pathways is the key phenomenon underpinning chronic fatigue syndrome (CFS): intracellular inflammation, with an increased production of nuclear factor kappa beta (NFkappabeta), cyclo-oxygenase-2 (COX-2) and inducible NO synthase (iNOS); and damage caused by O&NS to membrane fatty acids and functional proteins. These IO&NS pathways are induced by a number of trigger factors, for example psychological stress, strenuous exercise, viral infections and an increased translocation of LPS from gram-bacteria (leaky gut). The 'psychosomatic' symptoms experienced by CFS patients are caused by intracellular inflammation (aches and pain, muscular tension, fatigue, irritability, sadness, and the subjective feeling of infection); damage caused by O&NS (aches and pain, muscular tension and fatigue); and gut-derived inflammation (complaints of irritable bowel). Inflammatory pathways (monocytic activation) are also detected in somatizing disorder. SUMMARY: 'Functional' symptoms, as occurring in CFS and somatization, have a genuine organic cause, that is activation of peripheral and central IO&NS pathways and gut-derived inflammation. The development of new drugs, aimed at treating those disorders, should target these IO&NS pathways. PMID- 19127707 TI - Did I do that?! PMID- 19127708 TI - Struggling for a new self: in-depth interviews with 21 patients with hyperacusis after an acute head trauma. AB - The aim of the study was to gain a better understanding of what suffering from hyperacusis means. Twenty-one out-patients referred to a hearing clinic due to persisting hypersensitivity to sounds after an acute head trauma were interviewed. Before the head trauma, they had lived an independent, active and externally oriented life. As a consequence of the trauma, the patients became psychosocially vulnerable and suffered from a variety of symptoms: hypersensitivity to sounds, difficulties with concentrating and remembering, increased anxiety and sensitivity to stress. Structuring everyday life and using a variety of coping strategies was necessary for reducing life stress and symptoms in the process of adjusting to the changed life situation. PMID- 19127709 TI - Anger experience and anger inhibition in sub-populations of African American and European American older adults and relation to circulatory disease. AB - This study examined ethnic differences in the link between anger experience and anger inhibition and that of circulatory disease (CD). To ascertain the effects of anger inhibition in older persons, health data from groups of African American, African Caribbean, Eastern European and European American adults were collected. Experienced anger and anger inhibition were significant predictors of CD only for the African American group and the relation between experienced anger and CD was mediated by anger inhibition. The data suggest that cultural factors play a role in the development of an anger-inhibitory style and that this trait may pose a serious risk factor for circulatory disease. PMID- 19127710 TI - Stress, burnout and coping: differences between women with coronary heart disease and healthy matched women. AB - Stress is becoming more significant for women, along with the increasing number of women in the workforce. The present study compared women with respect to burnout and coping abilities, and related to the impact of educational level on differences in coping strategies. Women with coronary heart disease reported a higher level of burnout and had the highest scores demonstrating lack of coping, which indicates lesser coping abilities. Differences concerning strain reduction, self-control and emotional distancing are discussed in terms of living conditions. We also discuss that in order to optimize the outcome of rehabilitation and prevention we need more research on women, of women and especially from women's point of view. PMID- 19127711 TI - Translating exercise intentions into behavior: personality and social cognitive correlates. AB - The purpose of this study was to detail the variability found in the exercise intention-behavior relationship and investigate social cognitive and personality correlates of successful intention translation. Participants were 300 undergraduate students who completed measures of exercise social cognition (theory of planned behavior), personality (five-factor model) and a two-week follow-up of exercise behavior. Results suggested intention translation at a frequency of zero was significantly more successful than intending to exercise at all other weekly frequencies. Moreover, intending to exercise one or two times per week resulted in better intention translation than intending to exercise four or more bouts per week. Discriminant function analysis and follow-up F-tests found instrumental attitude, affective attitude and perceived behavioral control (PBC) discriminated between nonintenders, unsuccessful intenders and successful intenders. Further, extroversion predicted unsuccessful intenders versus successful intenders. Results underscore the importance of attitude, PBC and extroversion as action control constructs in the exercise domain. PMID- 19127712 TI - Comparing symptoms of chronic fatigue syndrome in a community-based versus tertiary care sample. AB - Chronic Fatigue Syndrome (CFS) is a debilitating condition characterized by six or more months of prolonged or relapsing, unexplained fatigue of new or definite onset and at least four of eight associated somatic and cognitive symptoms. Almost all studies of samples with patients with CFS have relied on referrals from physicians or health facilities. Underserved minorities, who not only tend to manifest higher levels of chronic illness, but are also less likely to seek and receive adequate medical care, have not been adequately represented in these studies. The present study compared two groups of individuals with CFS, one from a community-based sample and another from a tertiary-based sample. Findings indicate that patients with CFS from tertiary care settings have a higher frequency of symptoms than those in the general population who have CFS. PMID- 19127713 TI - The role of coping appraisal in reactions to fear appeals: do we need threat information? AB - Fear appeal theory postulates a multiplicative relationship between threat information and coping appraisal on fear control (e.g., avoid thinking about breast cancer to reduce feelings of fear) and danger control (e.g. motivation to perform monthly breast self-examination to detect breast cancer). That is, the effects of threat information on measures of both coping modes are moderated by beliefs about one's ability to cope with the presented threat. However, in the present study no support for an interaction between threat information and coping appraisal was found on both coping modes. Instead threat information seems to contribute positively to fear control and not to danger control, whereas coping appraisal contributed positively to danger control and negatively to fear control. The results thus suggest that we might do without threatening information in persuading people to adopt health-promoting behaviours. PMID- 19127714 TI - Determination of amino acid composition of soybeans (Glycine max) by near infrared spectroscopy. AB - Calibration equations for the estimation of amino acid composition in whole soybeans were developed using partial least squares (PLS), artificial neural networks (ANN), and support vector machines (SVM) regression methods for five models of near-infrared (NIR) spectrometers. The effects of amino acid/protein correlation, calibration method, and type of spectrometer on predictive ability of the equations were analyzed. Validation of prediction models resulted in r2 values from 0.04 (tryptophan) to 0.91 (leucine and lysine). Most of the models were usable for research purposes and sample screening. Concentrations of cysteine and tryptophan had no useful correlation with spectral information. Predictive ability of calibrations was dependent on the respective amino acid correlations to reference protein. Calibration samples with nontypical amino acid profiles relative to protein would be needed to overcome this limitation. The performance of PLS and SVM was significantly better than that of ANN. Choice of preferred modeling method was spectrometer-dependent. PMID- 19127715 TI - Chemical structure of wood charcoal by infrared spectroscopy and multivariate analysis. AB - In this work, the effect of temperature on charcoal structure and chemical composition is investigated for four tree species. Wood charcoal carbonized at various temperatures is analyzed by mid infrared spectroscopy coupled with multivariate analysis and by thermogravimetric analysis to characterize the chemical composition during the carbonization process. The multivariate models of charcoal were able to distinguish between species and wood thermal treatments, revealing that the characteristics of the wood charcoal depend not only on the wood species, but also on the carbonization temperature. This work demonstrates the potential of mid infrared spectroscopy in the whiskey industry, from the identification and classification of the wood species for the mellowing process to the chemical characterization of the barrels after the toasting and charring process. PMID- 19127716 TI - Identification of (12-ricinoleoylricinoleoyl)diricinoleoylglycerol, an acylglycerol containing four acyl chains, in castor (Ricinus communis L.) oil by LC-ESI-MS. AB - An acylglycerol (AG) containing four acyl chains, (12 ricinoleoylricinoleoyl)diricinoleoylglycerol (RRRR), was positively identified for the first time in a natural source in castor oil using electrospray ionization tandem mass spectrometry (ESI-MS/MS). HPLC-purified RRRR from castor oil was subjected to ion trap and high-resolution ESI-MS/MS. The precursor and fragment ions of [RRRR + Na]+ showed the expected masses, and the sodiated fragment ions of both diacylglycerols and fatty acids were detected. Because fragment ions of fatty acids from [AG + NH4]+ adducts cannot be detected by ESI MS/MS, [AG + Na]+ adducts are more informative. Radiolabeled triricinolein (RRR) was incorporated into RRRR in castor microsomes, indicating that RRRR is biosynthesized in castor bean. This newly identified and biosynthesized RRRR represents a new AG subclass of tetra-acylglycerols (or acylacyldiacylglycerol). PMID- 19127717 TI - Production of stilbenoids and phenolic acids by the peanut plant at early stages of growth. AB - The peanut plant (Arachis hypogaea) is known to produce stilbene phytoalexins as a defensive response to fungal invasion; however, the distribution of phytoalexins among different organs of the peanut plant at early stages of growth under axenic conditions has not been studied. Axenic plants produced a stilbenoid, resveratrol, as well as soluble bound and free phenolic acids, including 4-methoxycinnamic acid, which is reported in peanuts for the first time. Neither resveratrol nor phenolic acids were found in the root mucilage; the prenylated stilbenes were restricted to the mucilage and were not found in other organs of the peanut plant. These findings may lead to a better understanding of the defensive role of peanut stilbenes and phenolic acids. PMID- 19127718 TI - Molecular orbital basis for yellow curry spice curcumin's prevention of Alzheimer's disease. AB - It is demonstrated by using high-level ab initio computations that the yellow curcumin pigment, bis-(4-hydroxy-3-methoxyphenyl)-1,6-diene-3,5-dione, in the east Indian root plant turmeric (Curcuma longa) exhibits unique charge and bonding characteristics that facilitate penetration into the blood-brain barrier and binding to amyloid beta (Abeta). Alzheimer's disease is caused by Abeta accumulation in the brain cells combined with oxidative stress and inflammation. Consistent with the recent experimental work by Cole and co-workers (Yang, F., et al. J. Biol. Chem. 2004, 280, 5892-5901) that demonstrates curcumin pigment's binding ability to Abeta both in vivo and in vitro, it is shown here that curcumin possesses suitable charge and bonding features to facilitate the binding to Abeta. In addition, curcumin's anti-inflammatory and antioxidant properties are also attributed to electronic and structural features. It is shown that the presence of an enolic center and two phenolic polar groups separated by an essentially hydrophobic bridge of a conjugated network provides both hydrophobic and hydrophilic features to the curcumin pigment, thereby facilitating penetration into the blood-brain barrier through the former property and then binding to Abeta oligomer through the latter property. Both density functional and Moller-Plesset perturbation (MP2) computations have been carried out on the curcumin pigment to obtain fully optimized geometries in the gas phase and aqueous solution and also the atomic charges. Different isomers (keto and enol forms) have been considered to show that the enol form is the most favored and has all of the properties for an ideal antioxidant with also features to penetrate the blood-brain barrier and to bind to Abeta. This is demonstrated with natural bond charges, highest occupied and lowest unoccupied molecular orbitals, dipole moments, and Laplacian plots. The computed ionization potential and electron affinity show that curcumin has a low molecular hardness and thus has a propensity to dissociate its phenolic -OH, and the resulting charge undergoes delocalization throughout the structure, resulting in excitonic features. This feature seems to be also important for its binding capability to human proteins such as human serum albumin and Abeta. PMID- 19127719 TI - Antifungal and antibacterial activities of Mexican tarragon (Tagetes lucida). AB - Mexican tarragon (Tagetes lucida Cv. Asteraceae: Campanulatae) is an important, nutritious plant and an effective herbal medicine. Seven coumarins, 7,8 dihydroxycoumarin (4), umbelliferone (7-hydroxycoumarin) (5), scoparone (6,7 dimethoxycoumarin) (7), esculetin (6,7-dihydroxycoumarin) (11), 6-hydroxy-7 methoxycoumarin (12), herniarin (7-methoxycoumarin) (13), and scopoletin (6 methoxy-7-hydroxycoumarin) (14), and three flavonoids, patuletin (18), quercetin (19), and quercetagetin (20), were isolated from CH2Cl2 and MeOH extracts from aerial parts of T. lucida. In addition, 6,7-diacetoxy coumarin (15), 6-methoxy-7 acetylcoumarin (16), and 6-acetoxy-7-methoxycoumarin (17) derivatives were synthesized. 8-Methoxypsoralen (1), 8-acetyl-7-hydroxycoumarin (2), 7,8-dihydroxy 6-meth-oxycoumarin (3), 6,7-dimethoxy-4-methylcoumarin (6), 5,7-dihydroxy-4 methylcoumarin (8), 4-hydroxycoumarin (9), 4-hydroxy-6,7-dimethylcoumarin (10), naringenin (21), glycoside-7-rhamnonaringin (22), and rutin (23) were commercially obtained (Sigma-Aldrich). All of these compounds and extracts (M1 and M2) were assayed against bacteria and fungi. The antibacterial activity was determined on Bacillus subtilis, Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Salmonella typhi, Salmonella sp., Shigella boydii, Shigella sp., Enterobacter aerogenes, Enterobacter agglomerans, Sarcina lutea, Staphylococcus epidermidis, Staphylococcus aureus, Yersinia enterolitica, Vibrio cholerae (three El Tor strains, CDC-V12, clinic case, and INDRE-206, were obtained from contaminated water), and V. cholerae (NO-O1). The evaluated fungi were Aspergillus niger, Penicillium notatum, Fusarium moniliforme, Fusarium sporotrichum, Rhizoctonia solani, and Trichophyton mentagrophytes. The most active compounds against Gram-positive and -negative bacteria were the dihydroxylated coumarins 3 and 4. In addition, 2-4, 6, 7, and 11 showed an interesting activity against V. cholerae, a key bacterium in the contaminated water; 2-4 were the most active. Coumarins were the most effective compounds against Gram-negative bacteria. The extract MeOH/CH2Cl2 (1: 4) M2 at 0.4 microg/disk inhibited the growth of E. coli and P. mirabilis (40%), K. pneumoniae (31.1%), Salmonella sp. (35.5%), and Shigella sp. (0%) at 72 h of culture. The dimethoxy compounds 6 and 7 showed a strong activity against fungal strains, especially T. mentagrophytes and R. solani (100% of inhibition at 125.0 and 250.0 microg/mL, respectively). PMID- 19127720 TI - Inhibition of lipid accumulation and lipid body formation in oleaginous yeast by effective components in spices, carvacrol, eugenol, thymol, and piperine. AB - We screened natural organic compounds, which affected the lipid accumulation and the lipid body formation in oleaginous yeast, Lipomyces starkeyi, generating large lipid bodies. We found that four natural components in spices, carvacrol, thymol, eugenol, and piperine, inhibited the lipid accumulation at concentrations of 20-50 mg/L with a slight growth inhibition. The inhibitory effects were quantitatively represented by the total lipid accumulation amount, the triacylglycerol accumulation amount, and the average lipid body size. At 50 mg/L, the effects of these compounds were not identical and exhibited 11-37% decrease in lipid amount and 15-21% decrease in lipid body size with 13-39% decrease in cell growth. The inhibitory effect of these compounds lead to 30-69% decrease in triacylglycerol accumulation without any additional accumulation of its intermediates, suggesting that they will suppress the total carbon inflow into the triacylglycerol biosynthesis. PMID- 19127721 TI - Modulation of aromatase activity by diet polyphenolic compounds. AB - Estrogens are involved in physiological actions related to reproduction, body fat distribution, and maintenance of bone mass and are also related to the pathogenesis of estrogen-dependent cancers. The aim of this work was to study the effect of polyphenols on estrogen synthesis. The effect of polyphenols and polyphenolic-rich beverages on aromatase activity was tested in JAR cells (a choriocarcinoma cell line) through the tritiated water release assay. Some of the tested polyphenols inhibited estrogen production, chrysin being the most potent. Additionally, we observed that red wine, alcohol-free red wine, green tea, and black tea (200 microL/mL) significantly decreased aromatase activity. No effect on aromatase expression, as assessed by western blotting and RT-PCR, has been detected after 24 h of treatment with any of the flavonoids under study. In conclusion, polyphenols are able to modulate aromatase activity and, consequently, estrogen synthesis. The knowledge of such interference may help to clarify some of the biological properties attributed to polyphenols and may be useful in prevention/treatment of estrogen-dependent disorders. PMID- 19127722 TI - Influence of dietary antioxidants on polyphenol intestinal absorption and metabolism in rats. AB - The ingestion of complex foodstuff implicates the simultaneous presence along the digestive tract of several antioxidants micronutrients. This work aimed to determine if the interactions that may occur at the digestive level between polyphenols and other antioxidant micronutrients may modulate the fate of polyphenols in the splanchnic area. This study was conducted in a rat in situ intestinal perfusion model. Polyphenols (gallic and caffeic acids, catechin, and naringenin) were perfused with or without an antioxidant cocktail. For gallic acid, a significant reduction (-20%) of its net transfer through the brush border associated with a drastic decrease of its intestinal secretion of conjugates ( 90%) was observed. By contrast, the transfer of catechin through the brush border increased (18%) and the secretion of its conjugates was twice in the presence of the antioxidants. Perfused polyphenols largely differed in their respective biliary secretion, but these fluxes were not modified by the presence of the antioxidant cocktail. Finally, the simultaneous presence in the small intestine of polyphenols and other dietary antioxidants never affects polyphenol splanchnic metabolism but may modify their intestinal transport without noticeable consequence on their final availability for peripheral tissues. PMID- 19127723 TI - Toxicity of bisabolangelone from Ostericum koreanum roots to Dermatophagoides farinae and Dermatophagoides pteronyssinus (Acari: Pyroglyphidae). AB - The acaricidal activity of materials derived from the roots of Ostericum koreanum (Apiaceae) toward adults of Dermatophagoides farinae and Dermatophagoides pteronyssinus was examined by direct contact and vapor phase toxicity bioassays. Results were compared with those of three acaricides: benzyl benzoate, dibutyl phthalate, and N,N-diethyl-m-toluamide (DEET). The active principle was identified as the sesquiterpenoid bisabolangelone by spectroscopic analysis. In fabric-piece contact toxicity bioassays using adult D. farinae, bisabolangelone (1.88 microg/cm2) was more toxic than benzyl benzoate (11.91 microg/cm2), DEET (62.20 microg/cm2), or dibutyl phthalate (79.54 microg/cm2), based on 24-h LD50 values. Against adult D. pteronyssinus, bisabolangelone (1.79 microg/cm2) was similarly more active than benzyl benzoate (9.65 microg/cm2), DEET (64.45 microg/cm2), and dibutyl phthalate (77.79 microg/cm2). In vapor phase toxicity tests with both mite species, bisabolangelone was equitoxic in closed versus open containers. These results indicate that bisabolangelone was largely toxic through contact action. Bisabolangelone merits further study as a potential contact acaricide or lead for the control of house dust mites. PMID- 19127724 TI - Antibacterial and antioxidant activities of quercetin oxidation products from yellow onion (Allium cepa) skin. AB - Four new quercetin-derived oxidation products (1-4) and lunularin-4-O-beta-D glucoside (5) were isolated from a water extract of onion (Allium cepa) skin, together with 17 other known compounds. Antibacterial assays for the isolated compounds showed that 2-(3,4-dihydroxyphenyl)-4,6-dihydroxy-2-methoxybenzofuran-3 one (1) presented selective activity against Helicobacter pylori strains and 3 (quercetin-8-yl)-2,3-epoxyflavanone (4) showed antibacterial activity against MRSA and H. pylori strains at the same time that it increased susceptibility of MRSA to beta-lactams. Evaluation of antioxidant activity against DPPH for the isolated compounds showed that the new derivative compounds (1-4) and 2,5,7,3',4' pentahydroxy-3,4-flavandione (6) are more active than quercetin. PMID- 19127725 TI - Alkali-soluble polysaccharides of Rhizoclonium riparium alga induce IL-1 gene expression via protein kinase signaling pathways. AB - Fortification of aquaculture foodstuff with various algae may improve the resistance of certain fish or shrimp to diseases and, as a routine procedure, has become ever more popular and, seemingly, important. Herein, we isolated certain alkali-soluble polysaccharides from a Rhizoclonium riparium alga (RASP), polysaccharides that can be separated into two different groups on the basis of the polysaccharide's molecular weight. Using gas chromatography-mass spectometry analysis, we found that the major monosaccharides constituting the higher molecular-weight group of RASP were galactose (41.99%), glucose (34.53%), xylose (20.24%), and mannose (3.24%). Using a murine-derived macrophage cell line J774A.1, we found that polysaccharide constituents of the higher molecular-weight group of RASP were able to induce interleukin-1beta (IL-1) gene expression via protein kinase-mediated signal transduction pathways. In essence, we found that c Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38), but not extracellular signal-regulated kinase (ERK), play an important role in the regulation of IL-1 gene expression in RASP-stimulated J774A.1 cells. To the best of our knowledge, this is the first occasion that polysaccharides from R. riparium have been demonstrated to exert immunomodulation properties by the induction of IL-1 within macrophages. Our current results provide support for the possible use of R. riparium as an additive to various food/foodstuff, to modulate the immune response of humans or certain animals. PMID- 19127726 TI - Biovalorization of friedelane triterpenes derived from cork processing industry byproducts. AB - Here, we describe the synthesis, bioactivity screening, and structure-activity relationships of various synthetic triterpenoids prepared from the cork processing byproducts friedelin (1) and 3-hydroxyfriedel-3-en-2-one (2) via oxidative procedures. The synthesis of compounds 2alpha-trimethylsiloxyfriedelan 3-one (17), friedelin-2,3-lactone (18), friedelin-3-oxime (19), and friedelin-3,4 lactam (20) is also described. We have studied the insecticidal and phytotoxic potential of these compounds, their selective cytotoxic effects on insect and mammalian cells, and their antiparasitic effects. Structural modifications of the A-ring of friedelin (1) improved its insecticidal activity with derivatives 5, 2,3-secofriedelan-2-al-3-oic acid (6), its acetylated derivative 6a, 3beta- and 3alpha-hydroxyfriedelane (9 and 10), 3alpha-hydroxyfriedel-2-one (11), 4beta hydroxyfriedel-3-one (16), the acetylated 10a, 3,4-secofriedelan-4-oxo-3-oic-acid (14), lactone 18, and the oxime 19 being stronger insecticides than the parent compound. Methyl-3-nor-2,4-secofriedelan-4-oxo-2-oic acid (12) and its acetylated derivative 12a also showed insecticidal activity in contrast to their inactive parent compound 2. The postingestive effects and cytotoxicity of these compounds suggest a multifaceted insecticidal mode of action. These structural modifications did not result in better phytotoxic agents than the parent compounds except for lactam 20 and yielded several moderately active antiparasite derivatives (seco acids 6, 12, 14, and 4beta-hydroxyfriedel-3-one 16) with cytotoxic effects on mammalian cells. PMID- 19127727 TI - Preparative isolation and NMR characterization of carboxypyranoanthocyanins. AB - Three pyranoanthocyanins, the 3-glucosides of 5-carboxypyranodelphinidin (2), 5 carboxypyranopetunidin (4), and 5-carboxypyranomalvidin (6), were produced by nucleophilic addition of pyruvic acid to a purified extract of black beans (Phaseolus vulgaris) containing a mixture of the 3-glucosides of delphinidin (1), petunidin (3), and malvidin (5). A Sephadex LH-20 column was used for preparative separation of both pyranoanthocyanins and anthocyanins. The optimum solvent used for isocratic separation was 20% methanol containing 0.5% trifluoroacetic acid. The chromatographic method applied on the pigment mixture yielded in a one-step separation pigments 1-6, with purities up to 98, 89, 99, 87, 55, and 81%, respectively. The structures of 2 and 4, which previously have been tentatively identified mainly by mass spectrometric data acquired from pigment mixtures in wine samples or modified blueberry extract, were confirmed in the present study by two-dimensional nuclear magnetic resonance spectroscopy. PMID- 19127728 TI - Selective separation of peptides contained in a rapeseed (Brassica campestris L.) protein hydrolysate using UF/NF membranes. AB - The ability of a charged UF membrane to fractionate the small peptides found in a rapeseed protein enzymatic hydrolysate, according to their charge characteristics, was investigated. The complexity of such a hydrolysate has required the setting up of technological alternatives to isolate the small peptides, to obtain a more efficient separation among the numerous peptide species. A preliminary step consisted of precipitation followed by filtration with a 3000 g/mol molecular weight cutoff (MWCO) membrane to obtain a solution concentrated in small peptides. The possibility of fractionating these small peptides by a charged 1000 g/mol MWCO membrane was investigated. The study enabled us to assess the contribution of electrostatic interactions during fractionation. The effect of pH and ionic strength on the peptide transmission was studied. The ionic strength contribution was considered by studying the effect on the selectivity of a desalting step by nanofiltration on a 500 g/mol MWCO membrane. Peptide transmission was lower at pH 9 than pH 4, and it was the lowest at pH 9 and low ionic strength. Ionic strength had a significant influence at pH 9 but showed no influence at pH 4. The amino acid analysis and capillary electrophoresis revealed that negatively charged (acid) peptides were found in lower proportions in the permeate. The opposite trend was observed for basic peptides, whereas neutral peptides were found in the same proportion in the retentate and the permeate. These results can be explained, according to the Donnan theory, by the existence of attractive and repulsive forces at the membrane-solution interface. Selectivity between basic and acid peptides was as high as 1.90 at pH 9 and low ionic strength. A rough sketch of a membrane-based process is proposed to fractionate rapeseed peptide mixtures. Results obtained were reproducible within 10%. PMID- 19127729 TI - Mutations on aromatic residues of the active site to alter selectivity of the Sulfolobus solfataricus maltooligosyltrehalose synthase. AB - Mutations Y290F, Y367F, F405Y, and Y409F located near subsite +1 were constructed in maltooligosyltrehalose synthase (MTSase) to alter the selectivity of the enzyme. These mutations were designed to evaluate the effects of hydrophobic interactions and/or hydrogen bondings on transglycosylation and side hydrolysis reactions. The catalytic efficiencies of Y290F MTSase for hydrolysis and transglycosylation reactions were only 6.6 and 5.6%, respectively, of those of wildtype MTSase, whereas the catalytic efficiencies of Y367F MTSase were decreased by about half. F405Y MTSase had similar catalytic efficiencies for transglycosylation and a somewhat lower catalytic efficiency for hydrolysis. Y409F MTSase had somewhat lower catalytic efficiencies for the transglycosylation and a similar catalytic efficiency for hydrolysis. Y290F and Y367F MTSases had large changes in delta(deltaG), suggesting that there are hydrogen bonds between the substrate and residues Y290 and Y367 of wild-type MTSase. Compared with wild type MTSase, F405Y MTSase had decreased ratios of hydrolysis to transglycosylation, whereas Y290F, Y367F, and Y409F MTSases had increased ratios. These results suggest that use of F405Y MTSase might result in a higher yield of trehalose production from starch when it replaces wild-type MTSase. PMID- 19127730 TI - Impact of fat and water crystallization on the stability of hydrogenated palm oil in-water emulsions stabilized by a nonionic surfactant. AB - The influence of (0-40 wt %) sucrose and (0 and 150 mmol/kg) sodium chloride on the physical properties of 20 wt % hydrogenated palm oil-in-water emulsions stabilized by 2 wt % Tween 20 after crystallization of the oil phase only or both the oil and water phases has been examined. Emulsion stability was assessed by differential scanning calorimetry measurements of fat destabilization after cooling-heating cycles and by measurements of mean particle size, percent destabilized fat, and percent free oil obtained from gravitational separation after isothermal storage (at -40 to +37 degrees C). At storage temperatures where the oil phase was partially crystalline and the water was completely liquid, the emulsions were unstable to droplet coalescence and oiling off because of partial coalescence. Both NaCI and sucrose increased the extent of partial coalescence in the emulsions. At storage temperatures where both oil and water crystallized, the emulsions were completely destabilized. The stability of the emulsions to freezing and thawing could be improved somewhat by adding sucrose (>20 wt %). Emulsions stabilized by whey proteins were shown to have better freeze-thaw stability than those stabilized by Tween 20, especially in the presence of sucrose. These results may help formulate food emulsions with improved freeze thaw stability. PMID- 19127731 TI - Functional fatty fish supplemented with grape procyanidins. Antioxidant and proapoptotic properties on colon cell lines. AB - This work shows the properties of grape procyanidins with additional anticarcinogenic properties for increasing the shelf life of functional seafood preparations. Galloylated procyanidins (100 ppm, 2.7 mean degree of polymerization, 25% galloylation) extended the shelf life of minced horse mackerel muscle stored at 4 degrees C more than 8 days compared to controls without addition of polyphenols. The levels of endogenous alpha-tocopherol, EPA, and DHA of fish muscle were also preserved after 10 days at 4 degrees C. Therefore, the presence of procyanidins increased the stability of a product based on minced fish muscle during cold storage and maintained its functionality associated with the presence of polyunsaturated fatty acids and alpha-tocopherol. In addition, grape procyanidins showed a significant capacity to induce apoptosis in colon cancer cells (HT29 cell line) while being inactive in noncancer control cells (IEC-6). Thus, the product based on fatty fish muscle supplemented with grape procyanidins appears to be a stable functional food offering the combined action of omega-3 fatty acids and natural polyphenols. PMID- 19127732 TI - Carnobacterium species: effect of metabolic activity and interaction with Brochothrix thermosphacta on sensory characteristics of modified atmosphere packed shrimp. AB - The importance of carnobacteria as spoilage microorganisms or potential protective cultures in food is not resolved, and little is known about their metabolism during growth in specific products. This study used chromatographic techniques including GC-MS and HPLC to evaluate the spoilage metabolism of Carnobacterium divergens, Carnobacterium maltaromaticum, and Carnobacterium mobile. Metabolic activity was studied in cooked and peeled modified atmosphere packed (MAP) shrimp at 5 degrees C as carnobacteria has been anticipated to contribute to spoilage of shrimp products. C. divergens and C. maltaromaticum caused sensory spoilage of shrimps and generated ammonia, tyramine, and various alcohols, aldehydes, and ketones. The effects of Carnobacterium species on the growth and metabolism of Brochothrix thermosphacta were also evaluated, but metabiosis between the two groups of bacteria was not observed. C. mobile and a specific cluster of C. maltaromaticum isolates (cluster L) did not cause sensory spoilage of shrimp. PMID- 19127733 TI - Lipid characterization of a high-stearic sunflower mutant displaying a seed stearic acid gradient. AB - In the seeds of the high-stearic sunflower mutant CAS-14 a gradient of increasing stearic acid exists from the embryo to the terminal extreme of the cotyledon. This gradient modifies the fatty acid composition of the total lipids, triglycerides, and phospholipids, which can best be appreciated in the triglycerides that pass from 16% stearic acid content in the embryo to 37.1% in the other extreme. This increase in the triglycerides occurs principally at the cost of the oleic acid content. The stearic content at position sn-2 of triglycerides is low, rising from 1.3% in the embryo to 3.4%, whereas at positions sn-1 +3 the stearic content is high and augments from 25.2% in the embryo to 41.0% at the other extreme. The molecular species of triglycerides are also modified; the disaturated triglycerides increase from 15.5 to 51.7%. Furthermore, for the first time in sunflower seeds, it is demonstrated that trisaturated triglycerides are present, arising probably due to a modification in the acyltransferase system that synthesizes the triglycerides. PMID- 19127734 TI - Chemical composition and in vitro antioxidant evaluation of commercial water soluble willow herb (Epilobium angustifolium L.) extracts. AB - Five commercially available water-soluble extracts prepared from the aerial parts of Epilobium angustifolium L. (Onagraceae) were screened for antioxidant-related properties in a battery of six in vitro assays. Total phenol content and qualitative-quantitative analyses were also carried out. The extracts demonstrated varying degrees of efficacy in each screen. Two extracts, denoted as nonfermented and Tver, were the most effective toward reducing iron(III), scavenging 1,1-diphenyl-2-picrylhydrazyl free radicals, inhibiting hydroxyl radical-catalyzed bovine brain-derived phospholipid degradation, and non-site- and site-specific hydroxyl radical-catalyzed 2-deoxy-D-ribose degradation. The activity profile of the extracts changed, however, when their iron(II) chelating ability was assessed. The nonfermented and Tver extracts were not as effective iron(II) chelators as the extract denoted as Lotos. All the extracts contained Folin-Ciocalteu-reactive substances, which was confirmed by the presence of predominantly polar phenolic analytes (i.e., hydroxylated benzoic acid derivatives and flavonoids). PMID- 19127735 TI - Content of phenolic compounds and free polyamines in black chokeberry (Aronia melanocarpa) after application of polyamine biosynthesis regulators. AB - The total contents of anthocyanins, flavonoids, and phenolics in 60 samples of black chokeberries (Aronia melanocarpa), after treating with catabolites of polyamine biosynthesis (KPAb) and ornithine decarboxylase inhibitor, were analyzed spectrophotometrically, and quercetin and free polyamine contents were analyzed by RP-HPLC with UV detection. The average total contents of the individual substances and phenolic subgroups in control berries were as follows (mg x kg(-1)): anthocyanines, 6408; flavonoids, 664; phenolics, 37,600; quercetin, 349. KPAb decreased total contents of anthocyanines and phenolics only slightly but significantly increased the content of flavonoids. This caused an important change in the abundance of flavonoids in the pigment complex. The absolute content of quercetin was increased, but its ratio to flavonoids content was decreased. Ornithine decarboxylase inhibitor had a markedly different effect as it significantly increased total content of anthocyanins and total phenolics, inhibited the total content of free polyamines, and stimulated the processes of saccharides transformation to phenolic pigments. PMID- 19127736 TI - Structural changes of polyacetylenes in American ginseng root can be observed in situ by using Raman spectroscopy. AB - The presented results show the special advantage of Raman spectroscopy in the investigation of polyacetylenes in American ginseng (Panax quinquefolium L.) roots. The compounds are measured directly in the plant tissue without any preliminary sample preparation. The polyacetylene signal is strong and well separated from other bands so the spectral impact of the surrounding biological matrix can be clearly distinguished. The Raman spectrum taken in situ from the fresh ginseng root revealed a characteristic polyacetylene key band at 2237 cm( 1) whereas in the spectrum obtained from dried root this band was shifted to about 2258 cm(-1). The latter is in good agreement with signals obtained from isolated standards, falcarinol (2258 cm(-1)) and panaxydol (2260 cm(-1)), occurring as predominant polyacetylenes in this species. The shift of the polyacetylene band observed in root extracts or at a certain stage of root drying indicates the molecular modification of polyacetylenes resulting from the loss of water. Furthermore, it was found that the process upon root hydration is reversible as the shift of polyacetylene band from 2258 to 2237 cm(-1) is observed. An explanation of this phenomenon can be an interaction of polyacetylene molecules with plant components in the presence of water molecules forming a stable entity in situ that is broken after dehydration (loss of water) of the fresh ginseng root. Application of the Raman mapping technique to ginseng roots of different size showed that the content of both main polyacetylenes decreases with increasing root size in accordance with quantitative high performance liquid chromatography data. PMID- 19127737 TI - Stereoselective synthesis and antifungal activities of (E)-alpha (methoxyimino)benzeneacetate derivatives containing 1,3,5-substituted pyrazole ring. AB - Thirteen novel (E)-alpha-(methoxyimino)benzeneacetate derivatives, the analogues of strobilurins, which contain two pharmacophoric substructures of the methyl (E) methoxyiminoacetate moiety and 1,3,5-substituted pyrazole ring, were stereoselectively synthesized. It was found that the coupling reaction could give stereoselectively (E:Z ca. 14:1) the key intermediate material (E)-methyl 2 (hydroxyimino)-2-o-tolyl acetate (2). An X-ray crystallographic structure determination was carried out in a representative product. The preliminary bioassays indicated that all of the compounds 1 showed potent fungicidal activity against Rhizoctonia solani, Botrytis cinereapers, Gibberella zeae, Physalospora piricola, and Bipolaris mayclis. PMID- 19127738 TI - Photochemical degradation of imazamox in aqueous solution: influence of metal ions and anionic species on the ultraviolet photolysis. AB - Imazamox [5-methoxymethyl-2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl) nicotinic acid] is one of the later pesticides of the imidazolinone family. The fate of imazamox has been studied upon UV irradiation. The degradation of a 10 mg L(-1) imazamox solution leads to pyridine derivatives, which remain in solution for 50-100 h. Most of the photoproducts occurring during the photodegradation have been characterized by means of liquid chromatography coupled with mass spectrometry. The degradation scheme is very similar to that observed for the analogous imazapyr pesticide [2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl) nicotinic acid]. Nevertheless, imazamox shows a greater stability than imazapyr. More, complexation interactions between imazamox and metal ions such as Cu2+ or Ca2+ increase the persistence of the pesticide. PMID- 19127739 TI - Identification of environmental factors limiting plant uptake of metaldehyde seed treatments under field conditions. AB - Slugs are serious pests of oilseed rape (canola) and wheat with most damage occurring just after sowing and seedling emergence. As an alternative to the use of bait pellets, molluscicidal seed treatments have been shown to protect seeds and seedlings from slug damage in laboratory and semi-field experiments. However, protection offered to plants in field trials was diminished and shortlived in comparison with laboratory experiments. To determine why field efficacy was reduced, we grew seedlings under a range of environmental conditions, with appropriate controls, that simulated differences between laboratory and field experiments. We then measured the metaldehyde content of plant seedlings using a previously unpublished methodology described herein, which, unlike previous methods, did not first depolymerize the metaldehyde to acetaldehyde. We confirmed that naturally abundant plant-derived acetaldehyde could not interfere with our measurements of metaldehyde, even if depolymerization took place within the column. Our data suggest that reduced field efficacy results from microbial breakdown and/or loss of active ingredient caused by percolating soil water. Once the seedlings had emerged, neither volatalization nor simulated rainwater reduced the metaldehyde content of seedlings. Our findings will help develop superior seed treatment formulations to overcome these constraints. PMID- 19127740 TI - Degradation of chlorothalonil in irradiated water/sediment systems. AB - Water/sediment systems were used to investigate partitioning behavior between waters and sediments, as well as the degradation of the fungicide chlorothalonil (CHT) in each matrix. Experiments were run in the light and dark simultaneously for 30 days in both creek and pond sediment systems. Of the total applied CHT, 87 88% dissipated from the water phase in both water/sediment systems within 1 day when irradiated by simulated sunlight. In contrast, 60-68% remained in the water at day 1 in the dark. Approximately 3-6 and 10-16% of the applied CHT was found in sediments under light conditions at day 1 and in the dark at day 3, respectively which are the highest amounts observed during the experimental period. CHT similarly behaved in irradiated water/sediments and sediment-free aqueous solutions, indicating that CHT primarily degraded by photodegradation rather than adsorption to sediment in the early stages of the experiment. 4 Hydroxychlorothalonil was detected only in water in the dark systems. Trichloro 1,3-dicyanobenzene and 3-cyano-2,4,5,6-tetrachlorobenzamide were also detected and identified with liquid chromatography-electrospray ionization-mass spectrometry. These results suggest that photodegradation is likely to be important to the dissipation of CHT in aqueous solutions and microbial degradation plays an important role for residues that would ultimately reside in sediment. PMID- 19127741 TI - Tea plant uptake and translocation of polycyclic aromatic hydrocarbons from water and around air. AB - This study, which aimed to investigate the capacity of tea plant uptake and translocation of polycyclic aromatic hydrocarbons (PAHs), was divided into two sections. One was to study tea plant root uptake of PAHs from water and translocation to leaves. The other was to research tea plant leaf uptake of PAHs from air. It was observed that tea plant roots and leaves could strongly accumulate PAHs from around the environment. The capacity of tea plant uptake and translocation of PAHs were found to be closely relative to the physical-chemical properties of PAHs. With the increase of IgK(ow) (octanol-water partition coefficient) of the PAHs, both root concentration factors and leaf concentration factors increased exponentially, while translocation factors from roots to leaves decreased exponentially. PMID- 19127742 TI - Kinetics of maltooligosaccharide hydrolysis in subcritical water. AB - The kinetics of the hydrolysis of maltooligosaccharides with a degree of polymerization (DP) of 3-6 in subcritical water was studied using a tubular reactor at temperatures between 200 and 260 degrees C and at a constant pressure of 10 MPa. The maltooligosaccharide disappearance and product formation at residence times shorter than 50 s could be expressed by first-order kinetics. The rate constants for the hydrolysis of each maltooligosaccharide were evaluated. There was a tendency that the exosite glucosidic bond was hydrolyzed faster than the endo-site one irrespective of the DP of the maltooligosaccharide. The hydrolysis of the maltooligosaccharides was consecutively preceded, and the time dependence of the hydrolysis for maltooligosaccharides with different DPs could be calculated by simultaneously solving the mass balance equations for all the possible saccharides. PMID- 19127743 TI - Selection, characterization, and CDR shuffling of naive llama single-domain antibodies selected against auxin and their cross-reactivity with auxinic herbicides from four chemical families. AB - Indoleacetic acid (IAA)-binding single-domain antibodies (sdAbs) were isolated from a naive phage-display library constructed from the heavy chain antibody repertoire of a Ilama. The highest-affinity sdAb isolated (CSF2A) had a K(D) of 5 20 microM for two IAA-protein conjugates and a K(D) of 20 microM for free IAA. This sdAb also bound to a synthetic auxin analogue, 1-naphthaleneacetic acid (NAA), and to six auxinic herbicides (K(D) values of 0.5-2 mM), but not to serotonin and tryptophan, which are structurally similar to IAA but have no auxinic activity. To understand how sdAb CSF2A binds IAA and to determine which complementary-determining region(s) (CDR) participate(s) most in binding IAA, CSF2A was shuffled with four other sdAb clones by staggered extension process (StEP). After panning against IAA, two shuffled sdAbs were found: sdAb CSB1A, which originated from three different parental clones, and sdAb CSE8A, derived from two parental clones. These shuffled sdAbs and CSF2A were each fused to the B subunit of the Escherichia coli verotoxin, resulting in the formation of the pentamerized sdAbs V2NCSB1A, V2NCSE8A, and V2NCSF2A, which were analyzed by surface plasmon resonance (SPR) along with the sdAbs previously isolated. The shuffled clones had affinity for IAA (20 microM) similar to that of the highest affinity parental clone CSF2A, but much lower affinity for the auxinic herbicides. CDR2 was instrumental in binding IAA, whereas hydrophobic CDR3 was important for binding the auxinic herbicides. A novel SPR methodology is also described for specific immobilization of pentamerized sdAbs, allowing determination of K(D) values of Ab interaction with underivatized, low molecular weight haptens. PMID- 19127744 TI - Flavor release from i-carrageenan matrix: a quantitative structure-property relationships approach. AB - We carried out a QSPR (quantitative structure-property relationships) approach to evaluate the influence of the chemical structure of aqueous matrixes over the partition coefficient between the gas phase and the matrix. The determination of the partition coefficient of flavor ingredients was performed by headspace analysis at equilibrium for both saline solution and i-carrageenan gel. Starting from an initial list of 90 descriptors, we selected 10 descriptors to perform equation generation by the GFA (genetic function approximation) method available in the Cerius2 package. The best obtained equations involve only five descriptors, which encode electronic properties of charges repartition on the molecule (Jurs-RNCS and Dipole-Z) and molecules' shapes (PMI-Y, Shadow-XY, and RadOfGyration), both for saline solution and for i-carrageenan gel. However, the best-fitting equation for carrageenan gel is obtained with a quadratic relation, suggesting that the effect of carrageenan polymers only modulates but does not change the interaction of aroma compounds with water molecules. PMID- 19127745 TI - Character impact odorants of fennel fruits and fennel tea. AB - The flavor of fennel fruits and fennel tea was examined by aroma extract dilution analysis of the respective dichloromethane extracts. In both fennel fruits and tea, trans-anethole, anisaldehyde, and trans-4,5-epoxy-2(E)-decenal showed high flavor dilution (FD) factors followed by fenchone, 1,8-cineole, (R)-alpha-pinene, estragole, and beta-myrcene. On the basis of these results, the odorants showing higher FD factors were quantified in tea as well as in fruits, and odor activity values (OAV) in tea were calculated by dividing the concentration of the compound by its recognition threshold in water. The highest OAV was found for trans anethole, followed by estragole, fenchone, 1,8-cineole, (R)-alpha-pinene, beta myrcene, and anisaldehyde. From a comparison of the concentrations of odorants in fruits and tea, trans-anethole and estragole showed similar extraction rates of approximately 10-15%, whereas the extraction rates for (R)-alpha-pinene, beta myrcene, and limonene were below 2%. In contrast to this, fenchone, camphor, linalool, and carvone showed higher extraction rates (26-50%), whereas the high apparent extraction rates of anisalcohol (393%) and vanilline (480%) were attributed to the formation from precursors. Sensory studies of aqueous models containing odorants in the amounts quantified in fennel teas revealed high similarity of the models with the tea and proved that all impact odorants had been identified in their correct concentrations. Further sensory experiments showed that estragole had no odor impact on the overall flavor of fennel tea, and, therefore, a reduction of estragole in fennel products would have no negative impact on their sensoric quality. In contrast to this, trans-anethole and fenchone were found to be character impact compounds of fennel. PMID- 19127746 TI - Inhibition of apple polyphenol oxidase activity by sodium chlorite. AB - Sodium chlorite (SC) was shown to have strong efficacy both as a sanitizer to reduce microbial growth on produce and as a browning inhibitor on fresh-cut apples in previous experiments. This study was undertaken to investigate the inhibitory effect of SC on polyphenol oxidase (PPO) and the associated mechanisms. The experiment showed that SC had a strong inhibition of apple PPO. The extent of inhibition was influenced by SC concentration and pH. Inhibition was most prominent at pH 4.5, at which approximately 30% of enzyme activity was lost in the presence of 10 mM SC, followed closely by that at pH 4.0 with a 26% reduction in PPO activity. The inhibition mode was determined using Dixon and Lineweaver-Burk plots, which established SC to be a mixed inhibitor of apple PPO for the oxidation of catechol. Preincubation of PPO with 8 mM SC for 8 min caused a maximum of 46% activity reduction compared to noninhibited control. However, preincubation of SC with catechol for 8 min resulted in no additional loss of PPO activity. These findings provide further evidence that the inhibition of PPO activity by SC is due to the inhibition of the enzyme itself rather than removal of the substrate. PMID- 19127747 TI - Feruloyl esterase utilization for simultaneous processing of nonwood plants into phenolic compounds and pulp fibers. AB - Wheat and oilseed flax straws were studied as raw material for papermaking. Two different aspects were investigated to valorize these agricultural byproducts: the capacity to recover some phenolic compounds and the use of the resulting cellulose fibers in papermaking. Straw phenolic compound composition was analyzed to determine the different accessible molecules and their available quantity. Ferulic acid, coumaric acid, vanillic acid, and vanillin were found in both wheat and oilseed flax straws. To enhance the release of these phenolic compounds, enzymatic treatments with feruloyl esterase and xylanase, two enzymes known for their role in lignin destructuration, were tested. These treatments increased the amount of phenolic compounds released, destructured hemicellulose and lignin, and improved the soda cooking conditions of pulps with the reduction of chemical charge need in the papermaking process. Phenolic compound production from this process could enhance the cost-effectiveness of papermaking from annual plants. PMID- 19127748 TI - Comparison of the structure and properties of liposomes prepared from milk fat globule membrane and soy phospholipids. AB - Liposomes were prepared from a milk fat globule membrane (MFGM) phospholipid fraction and from soy phospholipid material using a high-pressure homogenizer (Microfluidizer). The liposomes were characterized in terms of general structure, phase transition temperature, lamellarity, bilayer thickness, and membrane permeability. The liposomes prepared from the MFGM fraction had a significantly higher phase transition temperature, thicker membrane, and lower membrane permeability. These differences were attributed to different phospholipid compositions of the MFGM and soy phospholipid fractions. PMID- 19127749 TI - Gelatinization and solubility of corn starch during heating in excess water: new insights. AB - Starch gelatinization is associated with the disruption of granular structure causing starch molecules to disperse in water. This study was designed to examine starch granules as they were heated in water, and their resulting morphological, structural, and solubility traits. The results indicate that starch gelatinization is a more complex process than the previously suggested order-to disorder transition. The energy absorbed by the granules facilitates the rearrangement or formation of new bonds among molecules prior to the temperatures normally associated with the melting of amylopectin crystallites during gelatinization. It is also evident that amylose plays an important role during the initial stages of corn starch gelatinization. PMID- 19127750 TI - Human milk fat substitutes containing omega-3 fatty acids. AB - Structured lipids resembling human milk fat (HMF) enriched with omega-3 fatty acids were synthesized by enzymatic acidolysis reactions between tripalmitin, hazelnut oil fatty acids (FA), and omega-3 FA concentrate. Response surface methodology was used to model and optimize the incorporation of omega-3 FA and oleic acid into tripalmitin, in hexane, using immobilized sn-1,3-specific lipase, Lipozyme RM IM. The three factors chosen were substrate molar ratio, reaction temperature, and reaction time. Good quadratic models were obtained for the incorporation of eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) (response 1) and oleic acid (response 2) by multiple regression and backward elimination. The determination coefficient (R2) value for the models was 0.95. The adjusted R2 values were 0.91 and 0.92 for responses 1 and 2, respectively. The optimal conditions generated from the models for the targeted total EPA and DHA (5%) and oleic acid (40%) incorporation were 12.4 mol/mol, 55 degrees C, and 24 h for substrate ratio, temperature, and time, respectively. The model was verified, which led to the production of a HMF ingredient with 76.6% palmitic acid at the sn-2 position. PMID- 19127752 TI - Hen egg yolk low-density lipoproteins film spreading at the air-water and oil water interfaces. AB - Hen egg yolk is largely used as an ingredient in food emulsions due to its exceptional emulsifying properties. Low-density lipoproteins (LDL) are the main egg yolk constituents and the most important contributors to yolk emulsifying properties. To better understand the LDL adsorption mechanism and spreading at the interface, we extracted and studied LDL at different interfaces. At the air water interface, the LDL film isotherm presents three transitions, and two were identified by each lipid class present in LDL. The last transition should be due to apoproteins-lipids complexes. During LDL adsorption, the presence of apoproteins at the LDL surface and the neutral lipid core is necessary. At pH 3 and pH 7, LDL are disrupted and spread quasi-similarly at the air-water interface, contrary to the oil-water interface where LDL spread more at pH 7 than at pH 3. PMID- 19127751 TI - Spreading of monoglycerides onto beta-casein adsorbed film. Structural and dilatational characteristics. AB - The effect of monoglycerides (monopalmitin and monoolein) on the structural, topographical, and dilatational characteristics of betacasein adsorbed film at the air-water interface has been analyzed by means of surface pressure (pi)-area (A) isotherms, Brewster angle microscopy (BAM), and surface dilatational rheology. The static and dynamic characteristics of the mixed films depend on the interfacial composition and the surface pressure. At surface pressures lower than that for the beta-casein collapse (at the equilibrium surface pressure of the protein, pi(e)(beta-casein)) a mixed film of beta-casein and monoglyceride may exist. At higher surface pressures the collapsed beta-casein is partially displaced from the interface by monoglycerides. However, beta-casein displacement by monoglycerides is not quantitative at the monoglyceride concentrations studied in this work. The protein displacement by a monoglyceride is higher for monopalmitin than for monoolein and for spread than for adsorbed films. The viscoelastic characteristics of the mixed films were dominated by the presence of beta-casein in the mixture. Even at the higher surface pressures (at pi > pi(e)(beta-casein)) the small amounts of beta-casein collapsed residues at the interface have a significant effect on the surface dilatational properties of the mixed films. The structural, topographical, and viscoelastic characteristics of the mixed films corroborate the fact that protein displacement for monoglycerides is higher for spread than for adsorbed mixed films. PMID- 19127753 TI - Effects of copper on the antioxidant activity of olive polyphenols in bulk oil and oil-in-water emulsions. AB - The antioxidant activity and interactions with copper of four olive oil phenolic compounds, namely oleuropein, hydroxytyrosol, 3,4-dihydroxyphenylethanol-elenolic acid (1), and 3,4-dihydroxyphenylethanol-elenolic acid dialdehyde (2), in olive oil and oil-in-water emulsions stored at 60 degrees C were studied. All four phenolic compounds significantly extended the induction time of lipid oxidation in olive oil with the order of activity being hydroxytyrosol > compound 1 > compound 2 > oleuropein > alpha-tocopherol; but in the presence of Cu(ll), the stability of oil samples containing phenolic compounds decreased by at least 90%, and the antioxidant activity of hydroxytyrosol and compounds 1 and 2 became similar. In oil-in-water emulsions prepared from olive oil stripped of tocopherols, hydroxytyrosol enhanced the prooxidant effect of copper at pH 5.5 but not at pH 7.4. The stability of samples containing copper at pH 5.5 was not significantly different if oleuropein was present from that of the control. Oleuropein at pH 7.4, and compounds 1 and 2 at both pH values tested, reduced the prooxidant effect of copper. The lower stability and the higher reducing capacity of all compounds at pH 7.4 could not explain the higher stability of emulsions containing phenolic compounds at this pH value. However, mixtures containing hydroxytyrosol or oleuropein with copper showed higher 1,1-diphenyl-2 picrylhydrazyl radical scavenging activity at pH 7.4 than at pH 5.5. Moreover, the compound 2-copper complex showed higher radical scavenging activity then the uncomplexed compound at pH 5.5. It can be concluded that the formation of a copper complex with radical scavenging activity is a key step in the antioxidant action of the olive oil phenolic compounds in an emulsion containing copper ions. PMID- 19127754 TI - Identification and urinary excretion of metabolites of 5-(hydroxymethyl)-2 furfural in human subjects following consumption of dried plums or dried plum juice. AB - 5-(Hydroxymethyl)-2-furfural (I) is a major breakdown product occurring in solutions with high concentrations of fructose and glucose and is present in many fruit juices, in heat-sterilized parenteral solutions, and in baby cereals. The objective of this study was to characterize and identify 5-(hydroxymethyl)-2 furfural metabolites in human subjects following the consumption of dried plum juice and/or dried plums. Subjects were fasted overnight and blood and urine samples were obtained during the day following consumption. Subjects fed the dried plum juice and dried plums consumed 3944 micromol (497 mg) and 531 micromol (67 mg) of I, respectively. Four presumed metabolites of I were detected in the urine of subjects that consumed dried plum juice. They were tentatively identified using HPLC-MS/MS as (1) N-(5-hydroxymethyl-2-furoyl)glycine (III), (2) 5-hydroxymethyl-2-furoic acid (II), (3) (5-carboxylic acid-2-furoyl)glycine (IV), and (4) (5-carboxylic acid-2-furoyl)aminomethane (V). Total urinary excretion during the 6 h following the consumption of dried plum juice was 168, 1465, 137, and 75 micromoles on the basis of II as a standard for II, III, IV, and V, respectively. The estimated total recovery of I metabolites was 46.2% and 14.2% of the I dose during the first 6 h after consumption of dried plum juice and dried plums, respectively. I seems to be metabolized rapidly to glycine conjugates and other metabolites and excreted in the urine. PMID- 19127755 TI - Effects of dietary herring roe lipids on plasma lipid, glucose, insulin, and adiponectin concentrations in mice. AB - The aim of this study was to evaluate the effects of dietary Kazunoko (salted herring roe) lipids, which contain large amounts of cholesterol, phosphatidylcholine, and n-3 polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), on lipid and glucose metabolism in mice. Male Crlj:CD-1 (ICR) mice were fed one of four experimental diets which contained 6% lard, 1% Kazunoko lipids + 5% lard, 3% Kazunoko lipids + 3% lard, and 6% Kazunoko lipids for 12 weeks. Plasma total cholesterol, triacylglycerol, phospholipid, and glucose concentrations were significantly lower in the 3% and 6% Kazunoko lipid diet groups than in lard and 1% Kazunoko lipid diet groups (p < 0.05). Plasma adiponectin concentrations of mice fed the 6% Kazunoko lipid diet were higher than those of animals fed the lard diet group. These results suggest that EPA and DHA rather than cholesterol in the Kazunoko lipids influence the plasma total cholesterol level. The constituent Kazunoko lipids may not only decrease the levels of plasma lipids but also decrease glucose concentrations by enhancing plasma adiponectin levels in mice. PMID- 19127756 TI - Production of metabolites from the Penicillium roqueforti complex. AB - Penicillium roqueforti comprises three accepted species: P. carneum, which is associated with meat, cheese, and bread; P. paneum, associated primarily with bread and silage; and P. roqueforti, which is associated with various processed foods and silage. This paper reports the use of HPLC-MS and HPLC-NMR to investigate the metabolites of silage-derived strains from two areas where silage toxicoses are regularly observed (Scandinavia and eastern Canada). Only modest differences were seen between the metabolites produced by strains from Canada and Scandinavia; however, silage strains of P. paneum isolated from Quebec were poor producers of patulin. This paper reports for the first time the production of festuclavine from P. paneum. This may be important as a possible explanation for the ill thrift observed when this species is dominant in poorly ensiled materials fed to dairy cows. PMID- 19127757 TI - Massachusetts model seen as health care reform guide. PMID- 19127758 TI - Fibromyalgia coverage uneven despite recent drug approvals. PMID- 19127759 TI - Compensation monitor. Risk contracts help medical groups' bottom line. PMID- 19127760 TI - Countdown nears for launch of ICD-10. PMID- 19127761 TI - Price hikes spur Part D debate. PMID- 19127762 TI - Health insurers well positioned, but must plan for cost pressure. PMID- 19127763 TI - Home, (not so) sweet, medical home. PMID- 19127764 TI - Prediabetes: prevalent and preventable but underdiagnosed and undertreated. PMID- 19127766 TI - A few specialty drug prices fall--all generics. PMID- 19127765 TI - Nausea and vomiting of pregnancy: cost effective pharmacologic treatments. AB - Nausea and vomiting of pregnancy (NVP) can range from morning sickness to moderate NVP to hyperemesis gravidarum (HG). If it is left unmanaged, health plans may pay for expensive unproven outpatient therapies that are not necessary for treatment of simple morning sickness or moderate NVP. Meanwhile, patients with serious hyperemesis gravidarum whose treatment is delayed may suffer needlessly, ending up with multiple hospitalizations or emergency room (ER) visits. Two expensive, heavily marketed outpatient therapies with scant supportive evidence in the treatment of NVP have recently emerged and some health plans are providing coverage without a thorough review of the medical evidence or cost implications. Health plans may have an opportunity to save a significant amount and to improve member satisfaction by utilizing evidence-based knowledge of pharmacologic interventions that are driven, in order, by known safety, proven efficacy, and cost effectiveness. PMID- 19127767 TI - Transdermal patch tailors antinausea treatment. PMID- 19127768 TI - Managed care outlook. Rising deductibles for Part D beneficiaries. PMID- 19127769 TI - [Knockdown and overexpression of miR-219 lead to embryonic defects in zebrafish development]. AB - MicroRNAs (miRNAs) are important post-transcriptional modulators of gene expression, which have been implicated in many physiological and pathological processes. In this study, we found that miR-219 expressed since early segmentation stages (16s) in spinal cord and mid-, hindbrain in zebrafish using Northern blotting and whole mounts ISH technologies. Moreover, knockdown or overexpression of miR-219 led to specific embryonic defects. Furthermore, TUNEL assay showed that overexpression of miR-219 induced significant cell apoptosis in the head and tail of zebrafish. Our study suggested that miR-219 may play an important role in zebrafish embryonic development. PMID- 19127770 TI - [Fret-based single-molecule probes for monitoring induced activation of Rac, Cdc42 signaling pathways in living cells]. AB - Rho GTPases, including Rac1, Cdc42, play a critical role in the regulation of a variety of cellular processes such as cell morphology, cell migration, transcriptional activation and gene expression. We constructed several FRET-based single-molecule probes containing red fluorescent protein dsRed1, cyan fluorescent protein ECFP, the GTPase binding domain of the effector, Pak1 or N WASP, and Rac1 or Cdc42. Rac1 and Cdc42 signaling pathways were activated in transfected cells by the inducer, insulin or bradykinin respectively. In vitro fluorescent spectroscopy assays showed that FRET phenomena were observed in transfected NIH3T3 and Hela cells. For all 3 signaling pathways in NIH3T3 cells, the values of FRET efficiency reached the highest after induction for 5 min, but the increasing extents of the values of FRET efficiency varied in 3 signaling pathways. The values of FRET efficiency decreased with the extention of the induction time, but differed significantly in the decreasing speed for the signaling pathways. Rac1 and Cdc42 activation assays indicated that Rac1 and Cdc42 were in the activated state (GTP-bound) in the induced cells. Their relative activated activities in the cells induced for different time were consistent with the values of FRET efficiency. The activated Rac1, Cdc42 signaling pathways led to the formation of lamelliopodia and filopodia in the transfected cells respectively. The results showed that these single-molecule probes could be used to directly monitor the spatial and temporal imaging of the induced activation of the signaling pathways in living cells. With these single molecule probes, the GEF or GAP activities of putative regulatory proteins for Rac1 and Cdc42 were analyzed and judged, thus greatly simplifying the currently used methods for identifying the regulatory proteins for Rho GTPases. PMID- 19127771 TI - [Expression of galanin and galanin receptors in neurogenesis regions of adult mouse brain and effect of galanin on the neural stem cell's differentiation]. AB - The neuropeptid galanin is widely expressed in the central nervous system and has a diverse range of physiological effects including food intaking, arousal/sleep, nociception and reproduction. In this study, expression of galanin and galanin receptors (GalR1 and GalR2) mRNA were identified not only in the neurogenisis regions including subventricular zone (SVZ), rostral migratory stream (RMS) and dentate gyrus (DG) of adult mice but also in the SVZ-derived neural stem cell (NSC) culture. Here, we also showed that the addition of galanin and GalR2 specific agonist Gal2-11 to wild-type or GALKO NSCs under differentiation condition significantly promote the neuritogenesis and increase the length of neurites on the betaIII-tubulin positive cells. This effect could be reduced by treatment of the galanin antagonist M35. These results indicate that galanin and its receptors might regulate neurite extension in differentiating neural stem cells and even participate in the development of the nervous system. PMID- 19127772 TI - [Studies on the megasporogenesis and microsporegenesis and the development of the female gametophyte and male gametophyte in Camptotheca acuminate]. AB - The ovarium is hypostasy in Camptotheca acuminate Decne.. It has a locule and an ovule. The ovule is pendulous, anatropous andunitegmic. The ovule of Camptotheca acuminate Decne. is pseudocrassinucellate ovule. The development of embryo sac is polygonum type. Cytokinesis during the meiosis of microspore mother cells is of simultaneous type. The arrangement of microspores in tetrad is tetrahedral and isobilateral. One-nucleate microspore is triangle. Maturity pollen is triangle, circular and square. This paper mainly studied the megasporogenesis and microsporogenesis, and studied the development of their female and male gametophyte in Camptotheca acuminate Decne., and preliminarily discussed the cause of the part pistil abortion in Camptotheca acuminate Decne. PMID- 19127773 TI - [Dicer interacts with Wig-1 protein]. AB - Dicer is a key dsRNase in RNA interference, and Wig-1 is a p53-induced zinc finger protein with dsRNA-binding activity. In this report, we demonstrated a direct interaction between Dicer and Wig-1. Dicer associated with Wig-1, which was expressed as a glutathione S-transferase fusion protein, in a dose-dependent manner by GST pull-down assay. In a reciprocal experiment, Wig-1 appeared to compete with GST-eIF2C1 for the binding of Dicer. Far-western analysis showed that Dicer could bind to GST-Wig-1 directly. Furthermore, it was found that Wig-1 could increase the dsRNase activity of Dicer in vitro. Our findings raise the possibility that Dicer and Wig-1 may play a common role in dsRNA-related gene regulation. PMID- 19127774 TI - Angiogenesis inhibition in vascular endothelial cells by terpenoid compounds from Bletilla striata is via apoptosis pathway. AB - The inhibitive effects on angiogenesis and the potential mechanism by terpenoids from Bletilla striata were investigated in the study. Terpenoids were separated and purified from Bletilla striata by extraction and chromatography. The terpenoid and its crude extracts were used to treat on blood vessel of chick chorioallantoic membrane (CAM), as well as human umbilical vein endothelial cell (HUVEC). Our results demonstrated that the crude extracts containing the terpenoids could inhibit CAM angiogenesis and pure terpenoid sample could prohibit HUVEC proliferation and induce HUVEC undergoing apoptosis, which showed an apparent apoptotic characterizations, such as cell shrinking, blebbing of cell membrane, nucleus cracking, chromatin condensation and margination, apoptotic body formation and DNA degradation. Therefore, the inhibition on angiogenesis by terpenoids from Bletilla striata is depended on the induction of apoptosis to endothelial cells. PMID- 19127775 TI - [Effects of granulosa cells treatments and follicular fluid on cleavage rate and blastocyst rate of bovine oocyte after in vitro fertilization and culture]. AB - Experiments were conducted to study the effects of granulosa cell (GC), follicular fluid and their interaction on cleavage rate and blastocyst rate of Bovine oocytes following in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). A total of 2178 oocytes were used for studies on maturation, fertilization and embryo development. The cleavage rate was affected significantly (P < 0.05) by GC, follicular fluid and the interaction of GC and follicular fluid. The blastocyst rate was affected significantly (P < 0.05) by GC, the blastocyst rate wasn't affected significantly by GC, follicular fluid and the interaction of GC and follicular fluid (P > 0.05). Effects of granulosa cell and follicular fluid and their interaction on the cleavage rate and the blastocyst rate in embryo cultured sort order was GC > follicular fluid > interaction of follicular fluid and GC. It was concluded that follicular fluid and granulosa cell monolayer (GCM) adding in TCM199 better support maturation of oocytes and development of embryos in Bovine. There were no differences in the cleavage rate and blastocyst rate, between the groups granulosa cell monolayer and incubating granulosa cell (which were incubated 10 min in incubator before oocytes being transferred) on co-culture system. Granulosa cell monolayer could be replaced by incubating granulosa cell on coculture system on Bovine oocytes IVM, IVF and IVC. PMID- 19127776 TI - [Chinese waxy elites SW70 and SW22 are revealed to be new multimutants of starch synthesis enzyme coding genes]. AB - Waxy maize (wx) is a type of spontaneous starch mutant as compared to wild type foodstuff maize (Wx). The mechanisms underlying waxy maize kernel development are intricate and diversified. Here we characterized the expression of 21 genes belonging to four families, i.e., ADP-glucose pyrophosphorylase (AGPase), starch synthase (SS), starch branching enzyme (SBE) and starch debranching enzyme (DBE) in the developing kernels of waxy maize inbred SW22, SW70 and relative wild type inbred 5003 at 1, 2 and 4 weeks after pollination. Dynamic expression pattern of a number of genes in developing kernels of SW22 were different from that in 5003 and SW70. Besides, obvious presence of wx transcripts in SW22 and SW70 were observed, though at the level lower than that in wild type 5003. Unexpectedly, the transcripts of Gbssllb and isoamylase-type DBE coding gene Iso2 were completely absent in SW22 and SW70. The above observation prompted the hypothesis that partial or complete loss-of-function of Wx, and/or there exist lose-of function of uncharacterized gene (s) important for amylose synthesis in SW22 such as GbssIIb and Iso2, which may account for the absence of amylose accumulation in SW22. PMID- 19127777 TI - [Embryological observation on the wide cross of rice and maize]. AB - In this report, we studied the fertilization and embryo development of a cross between rice and maize. The results showed that the pollen grains of maize could enter into the tissue of style of rice, and reached the embryo sac and released the sperms. Maize sperm might fuse with rice egg or polar nucleus and formed zygote or primary endosperm. But the embryo and endosperm could not be developed normally, and thus the underdeveloped hybrid seeds were formed. The reason was probably that the parental genomes could not coordinate well. This work provided a clue to solve the crossing obstacles between different species. PMID- 19127778 TI - Effects of zebra leaf 1--a new variegation mutation--on chloroplast development in rice. PMID- 19127779 TI - [Prokaryotic expression and CRT-binding activity analysis of Hevea HbCBF1 protein]. AB - CBF pathway is the most important pathway during the process of cold acclimation in plants. In previous study, we cloned a CBF like gene HbCBF1 from Hevea brasiliensis, which implied that CBF pathway might exist in this kind of tropical oriented tree. However, it was still unknown if this gene functioned as C-repeat binding factor. Therefore it was very important to analyze the C-repeat binding activity of HbCBF1. Using a prokaryotic expression system, the HbCBF1 was expressed and the soluble fusion protein was extracted under the optimized conditions. The C-repeat binding activity of HbCBF1 protein was analyzed by gel shift assay, which indicated that the fusion protein could bind to the COR15a probe efficiently, and this binding activity could be com-peted with by overloading non-labeled COR15a probe. An unlabeled mutated probe M1, which did not include the C-repeat sequence, could not compete with the binding activity of HbCBF1 to COR15a probe. These results indicated HbCBF1 fusion protein could bind to the C-repeat containing DNA fragment specifically in vitro. PMID- 19127780 TI - Study on the increasing expression of thrombospondin-1 (TSP-1) and transforming growth factor-beta (TGF-beta) in myocardium of diabetic rats. PMID- 19127782 TI - Outcome of second trimester amniocentesis in twin pregnancies at Songklanagarind Hospital. AB - OBJECTIVE: To evaluate the outcome of genetic amniocentesis in twin gestations at Songklanagarind Hospital. MATERIAL AND METHOD: This was a descriptive study that included all women with twin pregnancies who had a second trimester amniocentesis for chromosome study at the Maternal Fetal Medicine Unit, Department of Obstetrics and Gynecology, Songklanagarind Hospital from January 1998 through June 2006 to assess the outcome including risk of fetal loss in such cases. RESULTS: Advanced maternal age was the most common indication for amniocentesis. The success rate of cell culture was 100%. The fetal loss within 14 days after the procedure was 1.4%. CONCLUSION: The tendency of fetal loss after amniocentesis in twin pregnancies was higher than in singletons. Pre-procedure counseling personnel should be aware of this potential outcome, and be sure to inform the involved parents prior to the procedure. PMID- 19127781 TI - Effectiveness of auricular acupressure in the treatment of nausea and vomiting in early pregnancy. AB - OBJECTIVE: To evaluate the effectiveness of auricular acupressure in the treatment of nausea and vomiting in early pregnancy. MATERIAL AND METHOD: Ninety eight volunteer pregnant women with symptoms of nausea and vomiting in early pregnancy before 14 weeks gestation were enrolled. The participants were randomized into two groups: treatment group and control group. Each patient in the treatment group received magnet pellets, placed at both auricles. They were taught to start acupressure from the third to the sixth day. Outcome measurement was Rhodes index score, which describe the severity and frequency of nausea and vomiting in the form of a questionnaire. The patients from both groups were asked to complete and return the forms including the amount of anti-emetic drug taken. Mean Rhodes index score and total number of anti-emetic drug taken from day 4-6 were used to compare the treatment effect. Student's t test, Chi-square test and Mann-Whitney U test were used for statistical analysis. RESULTS: Ninety-one pregnant women who returned the questionnaires were evaluated. The Rhodes index scores of the treatment group were lower than that of the control group especially after day 4 to day 6 when the acupressure was started. However when comparing the mean score between the two groups, there were no statistically significant differences (p > 0.05). The total amount of anti-emetic tablets in day 4-6 after acupressure intervention was compared and there were no statistically significant differences (p > 0.05) between the groups. CONCLUSION: Auricular acupressure therapy in treatment of nausea and vomiting in early pregnancy may not relieve nausea and vomiting in early pregnancy and need further clinical research to confirm the effectiveness. PMID- 19127783 TI - Preimplantation genetic screening (PGS) in infertile female age > or = 35 years by fluorescence in situ hybridization of chromosome 13, 18, 21, X and Y. AB - INTRODUCTION: It is common in infertile couples that the female partner age > or = 35 years, that some of them require assisted reproductive technology (ART) for their treatment, it is also well known that in this female age group increases the chance of chromosome aneuploidy in offsprings. It is known that the antenatal diagnosis may have the ethical dilemma and psychological impact. Therefore, the preimplantation genetic screening (PGS) may have a role in this ART group. OBJECTIVE: The present study had the objective to compare the incidence of normal, abnormal embryos and also aneuploidy of each chromosome, i.e. 13, 18, 21, X and Y between 2 subgroups of age i.e. the age 35-39 years and 32-39 years vs. the age > or = 40 years in both female and male partners respectively. MATERIALS AND METHOD: This prospective study was performed in 20 infertile couples attending the Fertility Clinic at Thammasat University Hospital during the years 2006-2007 of which the female partner aged > or = 35 years had to use the ART. The PGS was performed by FISH technique with 5 probes to detect the 13, 18, 21, X and Y chromosomes. The comparative analysis was made between 2 subgroups of both female and male partner aged, as mentioned above in the incidence of normal, abnormal embryos and aneuploidy of each chromosome by Chi-square test and Fisher's exact test with statistical significance if p < 0.05. RESULTS: The abnormal embryos in the female partner age > or = 40 years were higher than those of the age 35-39 years (72.4% vs. 52.5%, p = 0.07) but with no statistical significance. No different results were obtained in the comparable male partner age groups (56.8% vs. 61.4%, p = 0.66). The normal female and male embryos in the female partner age 40 years were lower than those of the age 35-39 years (10.4% vs. 25.4%, p = 0.08 and 17.2% vs. 22.1%, p = 0.60 respectively) but with no statistical significance. The normal female and male embryos in the male partner age > or = 40 years and the age 32-39 years were also compared with no significant differences (20.5% vs. 20.5%, p = 1.00 and 22.7% vs. 18.2%, p = 0.60, respectively). The percentage of embryos with aneuploidy of chromosome 18 in the female partner age > or = 40 years was significantly higher than that of the age 35-39 years (72.0% vs. 45.0%, p = 0.003). The pregnancy rate in the presented PGS study was 12.5% but unfortunately was associated with a high abortion rate of 100%. CONCLUSION: It was found in the present study that the incidence of abnormal embryos trend to increase in the female partner aged > or = 40 years compared to the aged 35-39 years although with no statistical significance. However, the incidence of embryos with aneuploidy of chromosome 18 was higher in females aged > or = 40 years with statistical significance, whereas the male partner age had no impact on the abnormality or normality of the embryo. The abortion rate was very high (100%) probably may be due to inadequate choice of probes, inappropriate fixation technology and small sample size. However, the results obtained in this study indicate that the PGS should be considerably performed with strong indication only. PMID- 19127784 TI - Prenatal ultrasonographic findings in "trisomy 13". AB - OBJECTIVE: To evaluate the accuracy of prenatal ultrasonographic diagnosis in fetuses with trisomy 13. MATERIAL METHOD: The present study consisted of all fetuses diagnosed of trisomy 13 and delivered at Ramathibodi Hospital between 1997 and 2006. RESULTS: There were 15 cases of trisomy 13. Twelve cases (80.0%) were detected by prenatal ultrasonographic examination, and 3 cases (20.0%) were missed. Mean maternal age was 31.4 years old. Sixty-six percent were diagnosed in 2" trimester (mean 19.4 weeks). The earliest gestational age for detection was 12 weeks 6 days. The most common abnormal ultrasonographic findings were holoprosencephaly (46.7%), and facial defects (40.0%). CONCLUSION: The accuracy of prenatal sonographic diagnosis in trisomy 13 fetuses was 80%. The most sensitive prenatal ultrasonographic findings in trisomy 13 were holoprosencephaly and facial defects. PMID- 19127785 TI - Effect of nicotine polyestex gum on smoking cessation and quality of life. AB - OBJECTIVE: To determine the effectiveness and safety of the novel nicotine polyestex gum for smoking cessation, along with its impact on the quality of life (QOL). MATERIAL AND METHOD: A double-blind, placebo-controlled, randomized clinical trial was conducted on 43 smokers. All of them received either nicotine gum or placebo. Only those who could quit completely and continuously by the end of 3 months were considered total abstinence. QOL was also measured using WHO questionnaires. RESULTS: Treatment with nicotine polyestex gum resulted in significantly greater abstinence rate at 3 months compared with placebo (50% vs. 9%; p = 0.003). Adverse events were modest and not encountered more often than those seen in the placebo group. QOL at 3-months improved in both groups, but there was no statistically significant difference between the groups. CONCLUSION: Nicotine polyestex gum is effective and safe for smoking cessation. It is also associated with significant improvement in the QOL. PMID- 19127786 TI - Evaluation of sensitivity and specificity of enzyme-linked immunosorbent assay (ELISA) for detecting antidesmoglein 1 and 3 in Thai patients with pemphigus vulgaris and foliaceus. AB - OBJECTIVE: Pemphigus is an acquired autoimmune blistering skin diseases, of which pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are two major subtypes. A novel commercial enzyme-linked immunosorbent assay (ELISA) against Dsg1 and Dsg3 has been well established for diagnosis and prediction of disease activity in PF and PV. At present, the benefit of anti-Dsg 1 and anti-Dsg 3 IgG by ELISA in the diagnosis of pemphigus in Thai patients has never been reported. The objective of the present study is to evaluate the sensitivity and specificity of ELISA for detecting antidesmoglein 1 and 3 in Thai patients with pemphigus. MATERIAL AND METHOD: Retrospective review of anti-Dsg1 and anti-Dsg3 antibody ELISA test results from 48 serum samples collected from 27 patients with PV seven patients with PF and 14 controls. RESULTS: The sensitivity of Dsg1 and Dsg3 ELISA for all patients with PV was 64% and 77.8% respectively. When subgrouped into only PV patients with new diagnosis, the sensitivity of Dsg 1 and Dsg 3 ELISA increased to 85.7% and 100%. In all PF patients, the sensitivity of anti-Dsg 1 ELISA was 71.4% and 100% for newly diagnosed PF cases. Anti-Dsg 3 was not detected in the PF group. The specificity of ELISA for anti-Dsg 1 and anti-Dsg 3 in both types of pemphigus was 85.7% and 92.3% respectively. CONCLUSIONS: Dsg 1 and Dsg 3 ELISA is a simple, highly sensitive and specific test in Thai pemphigus patients with 100% sensitivity in the diagnosis of both new pemphigus vulgaris and foliaceus patients. PMID- 19127787 TI - Exhaled carbon monoxide level and smoking status in urban Khon Kaen adults. AB - BACKGROUND: Recent data has shown that a great number of Thai adults as well as people worldwide have died from smoking-related diseases. Measurements of exhaled carbon monoxide have been increasingly used to evaluate smoking status and cutoff levels of exhaled carbon monoxide and have been widely reported among other populations but not for the Thai people. OBJECTIVE: The purpose of the present research was to study the proper cutoff level for exhaled carbon monoxide for detecting smoking status among urban Thai adults residing in Khon Kaen province, Thailand as well as to study the effect of baseline characteristics in modifying the cutoff level of exhaled carbon monoxide. STUDY DESIGN: Cross-sectional analytic study. MATERIAL AND METHOD: The present study employed existing data whereby the study subjects comprised a total of 420 Thai adults, aged 15-70 years, residing in urban Khon Kaen province, Thailand during the year 2006. The data was obtained through interview and exhaled carbon monoxide measurement. The analyses employed descriptive, bivariate, and multivariable logistic regression. RESULTS: Findings from the final multivariable logistic regression model revealed good relation between exhaled carbon monoxide levels and tobacco smoking status. Other variables in the model included age-group and the interaction between exhaled carbon monoxide and age-group. Further analysis showed a greater odds ratio in the older age-group, with the odds ratios (95% CI) being 2.50 (1.87, 3.34) and 1.46 (1.31, 1.63) in the older (41-70 years) and younger (15-40 years) age-groups, respectively. In addition, proper cutoff of exhaled carbon monoxide for the older age-group was suggested as 7 ppm, while 8 ppm was more appropriate for the younger age-group. Based on the findings, a baseline characteristic for age modified cutoff level of carbon monoxide was established CONCLUSION: Differences in baseline characteristics should be considered in evaluating smoking status when choosing the cutoff level of exhaled carbon monoxide for any population. PMID- 19127788 TI - Hemoglobin E detection using PCR with confronting two-pair primers. AB - OBJECTIVES: To develop and apply the polymerase chain reaction with confronting two-pair primers (PCR-CTPP) for detection and identification of hemoglobin E (Hb E). MATERIAL AND METHOD: Fifty unrelated northern Thais were included in the present study. DNA was extracted from peripheral blood mononuclear cells and targeted to amplify by PCR-CTPP. The amplified product was analyzed and compared with the reference hemoglobin electrophoresis and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: The results validated a completely concordant among these three methods consisting of 74%, 24%, and 2% identified as normal, heterozygous, and homozygous Hb E type, respectively. CONCLUSION: Successful Hb E genotyping by PCR-CTPP was introduced. It allows for confirming and simultaneously detection with other thalassemia mutations. PMID- 19127789 TI - Normal reference range of serum insulin-like growth factor (IGF)-I in healthy Thai adults. AB - BACKGROUND: Serum insulin-like growth factor (IGF)-I level is growth hormone (GH) dependent and reflects GH secretion. Analysis of IGF-I is a component in the diagnosis of GH-related disorders and is going to be of interest in determining the risk of many disorders such as cancer or atherosclerosis. The diagnosis value of IGF-I is dependent on the establishment of an accurate reference ranges, which can be affected by parameters such as age, gender, ethnicity, medications, chronic illness, or assay methodologies. OBJECTIVE: To determine reference ranges of IGF-I for healthy Thai adults. MATERIAL AND METHOD: Eight hundred sixteen healthy Thai adults aged between 21-70 years were recruited in the present study. Serum IGF-I was measured by using immunochemiluminescent (ICMA; Roche, USA). Subjects were recorded by their age and gender groups. Data were presented in mean and +/- 2 standard deviation (SD). Correlation analysis between serum IGF-I and physical parameters including sex, age, weight, height, and body mass index (BMI) was also made. RESULTS: The present study demonstrated normal reference range of serum IGF-I by using mean +/- 2 SD value. The well-known age dependency of serum IGF-I levels was also revealed. Levels decreased with increasing age in both genders. The mean value of serum IGF-I was slightly higher in women at the age of 30-40 years compared with men in the same age group, but not statistically insignificant. In addition, serum IGF-I was found to correlate directly with the height and negatively with BMI. However, age-adjusted IGF-I level did not show correlation with these physical parameters. CONCLUSION: This reference range will be beneficial for using IGF-I assay as a tool in the diagnosis of GH function abnormalities in Thai subjects. PMID- 19127790 TI - Prevalence of dementia among population age over 45 years in Chiang Mai, Thailand. AB - OBJECTIVE: To determine the prevalence of dementia in Thai people with age 45 years and above. MATERIAL AND METHOD: This project used a cross sectional research design to study the prevalence of dementia in Chiang Mai. Door-to-door technique was assigned in condition with multi-stage probability random sampling to obtain subjects representing the population of Chiang Mai between Oct 2004 and Sep 2005. The researchers collected the data from the subjects aged 45 years and above. All subjects were located from every Amphurs of Chiang Mai. They were first screened with Thai Mini Mental State Examination (TMSE) and Thai Beck Depression Inventory (BDI). The subjects whose TMSE was less than 24 were assessed and diagnosed by a neurologist. Subjects who were determined as having dementia might be laboratory analyzed and classified based on DSM-IV and NINDS AIREN criteria. RESULTS: The authors enrolled 2,311 people and screened them with Batteries test. One thousand four hundred ninety two people qualified with 610 males and 882 females, whose mean age was 59.7 +/- 10.4 years. The authors found that among the 35 people with dementia, the mean age was 67.9 +/- 8.9 years (45 88 years). The prevalence of dementia among the study participants was 2.35%. In the present study, Alzheimer's disease was the most common type of dementia diagnosed (75.0%) and vascular dementia was the second most commonly diagnosed (12.5%). CONCLUSION: The prevalence of dementia in Chiang Mai was 2.35%, which does not differ from the previous study Alzheimer's disease was the most common type of dementia diagnosed. PMID- 19127791 TI - The validation of the disease-specific questionnaire for health-related quality of life in Thai patients with hemifacial spasm. AB - OBJECTIVE: To investigate the reliability and validity of the Thai version of disease-specific health-related quality of life instrument for patients with hemifacial spasm (HFS) as well as their response to botulinum toxin treatment. MATERIAL AND METHOD: A Thai version of HFS-30 has been developed with the permission of the author. Thirty patients with HFS were asked to complete this Thai HFS-30, the 6-point disability scale before treatment and between four and six weeks after botulinum toxin injections. Peak improvement (0-100%) was subjectively assessed by each patient between four and six weeks after injection. They were also asked to answer the existing Thai SF-36 questionnaire before treatment to test its correlation with Thai HFS-30. Another group of ten patients completed the questionnaire and then a second identical copy after a 2-week interval. The reliability, validity, and responsiveness were subsequently analyzed. RESULTS: The Thai HFS-30 showed a Cronbach's alpha coefficient of 0.78 and no significant difference of a test-retest reliability. The total content validity was 0.88 (range 0.5-1.0). There were good correlations between both the Physical and Mental Health parts of the Thai HFS-30 and Thai SF-36 (p < 0.05 and p < 0.01, respectively). The Thai HFS-30 also demonstrated a response to treatment similar to the 6-point disability scale and the peak improvement. CONCLUSION: The Thai version of HFS-30 is a valid, reliable, and sensitive to change instrument for disease specific health-related quality of life assessment. PMID- 19127792 TI - The Thai anesthesia incident monitoring study (Thai AIMS): an analysis of perioperative myocardial ischemia/infarction. AB - OBJECTIVES: To analyze the clinical course, outcome, contributing factors and factors minimizing the incidents of perioperative myocardial ischemia or infarction (PMI) from Thai AIMS study. MATERIAL AND METHOD: The present study was a prospective multicenter study. Data was collected from 51 hospitals in Thailand during a six-month period. The participating anesthesia provider completed the standardized incident report form of the Thai AIMS as soon as they found the PMI incident. Each incident was reviewed by three peer reviewers for clinical courses, contributing factors, outcome and minimizing factors of PMI. RESULTS: From the Thai AIMS incident report, the authors found 25 suspected PMI cases which was 0.9% of the 2,669 incidents reported in the present study. Most of the PMI occurred in elective cases (84%) and orthopedic procedures (56%). The majority of PMI was reported from the patients undergoing general anesthesia (72%). Suspected PMI occurred mostly during operations (56%). New ST-T segment change was detected in 92% of these patients. The most common immediate outcome of PMI was major physiological change (88%). The most common management effect of PMI was unplanned ICU admission (64%); the others were prolonged ventilatory support (12%) and prolonged hospital stay (16%). Four patients (16%) died after the suspected PMI. Most of the events occurred spontaneously and were unpreventable (80%). Patient factors (100%), anesthesia factors (72%), surgical factors (32%) and system factors (8%) were all judged as a precipitating factor for PMI. Human factors were the most common contributing factors which included poor preoperative evaluation, inexperience and improper decision. The three most common factors minimizing the adverse incidents included prior experienced, high awareness and experienced assistance. The recommended corrective strategies were guideline practice, quality assurance activity, improvement of supervision and additional training. CONCLUSION: Perioperative myocardial ischemia/infarction was infrequent but may be lethal. Patient factors were the most common precipitating cause. The morbidity and mortality could be reduced by high quality preoperative evaluation and preparation, early detection and appropriate treatment. Guideline practice, quality assurance activity, improvement of supervision and additional training were suggested corrective strategies. PMID- 19127793 TI - The Thai Anesthesia Incident Monitoring Study (Thai AIMS) of post anesthetic reintubation: an analysis of 184 incident reports. AB - OBJECTIVE: The present study was a part of the Multi-centered Study of Model of Anesthesia related Adverse Events in Thailand by Incident Report (The Thai Anesthesia Incident Monitoring Study or Thai AIMS). The objective of the present study was to determine the outcomes, contributory factors and factor minimizing incident. MATERIAL AND METHOD: The present study was a descriptive research design. The authors extracted relevant data from the incident reports on reintubation after planned extubation after general anesthesia with endotracheal intubation from the Thai AIMS database during the study period January to June 2007. The cases were extensively reviewed by 3 reviewers for conclusion of anesthesia directly and indirectly related reintubation. Comparative analysis between two groups was done. RESULTS: A total 184 incidents of extubation failure according to the definition were extracted in which 129 cases (70.1%) were classified as directly related to anesthesia and 55 cases (29.9%) were indirectly related to anesthesia. Oxygen desaturation occurred in 85.9% of cases while 90.2% of patients was reintubated within 2 hours after extubation. Hypoventilation (58.1%) was the commonest cause which led to reintubation directly related to anesthesia while upper airway obstruction (39.6%) was the commonest cause in the indirectly related anesthesia group. The proportion of preventable incident was 99.2% and 54.5% in directly and indirectly related anesthesia groups, respectively. Human factors particularly including lack of experience and inappropriate decision-making were considered in 99.2%, are directly related to anesthesia reintubation group. CONCLUSION: Extubation failure and reintubation was mostly related to anesthesia. Most of directly related to anesthesia group were considered as preventable. Human factors were also claimed as contributing factors. Quality assurance activity and improvement of supervision to improve experience and competency of decision making were suggested corrective strategies. PMID- 19127794 TI - Trauma care audit using Srinagarind hospital's audit filter. AB - BACKGROUND: To audit trauma care (including the mortality rate and obstacles faced by the authors) at Srinagarind (University) Hospital using the trauma audit filter. MATERIAL AND METHOD: Conduct a prospective, descriptive, study of trauma patients who received trauma medical care at Srinagarind Hospital, Khon Kaen University, Thailand, between January and May 2006. Srinagarind Hospitals trauma audit filter was used to audit trauma care. The audit filter comprised 14 criteria (i.e., 1) emergency medical service; 2) accident and emergency out patient service; 3) in-patient service). Any filter that generated a "Yes" response was investigated to find the reason(s). The obstacles and mortality were also recorded. RESULTS: The authors enrolled 3209 patients. The mortality rate was 0.5% (95% CI 0.3-0.8). Emergency medical service, accident and emergency out- and in-patient service were rated satisfactorily. The reported obstacles were lack of hospital beds, inappropriate locale for trauma care, financial process, admission process, and lack of equipment. CONCLUSIONS: Srinagarind Hospital's audit filter had the capability to audit trauma care. Overall trauma care at Srinagarind Hospital was satisfactory albeit improvements are needed. PMID- 19127795 TI - Retroperitoneoscopic nephrectomy in dialysis dependent patients and comparison with open surgery. AB - OBJECTIVE: To evaluate the surgical outcomes and morbidity of retroperitoneoscopic nephrectomy compared with open nephrectomy for dialysis dependent patients. MATERIAL AND METHOD: Between November 2002 and August 2007, 14 hemo or peritoneal dialysis patients underwent nephrectomy or nephroureterectomy at Siriraj Hospital. Of the 14 patients, seven were treated with retroperitoneoscopic nephrectomy and seven with open nephrectomy. A retrospective review and data were carried out. The patient factors, type of surgery, perioperative outcomes and complications were analyzed. RESULTS: There was no conversion rate in the retroperitoneoscopic group. The mean estimated blood loss, analgesic requirement and time before starting oral intake were lower in the retroperitoneoscopic group (141.4 +/- 95 versus 292.8 +/- 226 ml, 5.0 +/- 4.5 versus 7.6 +/- 1.9 mg and 14.5 +/- 16.1 versus 23.1 +/- 23.3 hours, respectively). On the other hand, the mean operative time in the retroperitoneoscopic group was longer than the open group but with no significant difference (177.14 +/- 51 versus 160.71 +/- 84 min, p = 0.521). Two patients in the open group required intraoperative blood transfusion. There were two complications. One patient developed a large retroperitoneal hematoma after retroperitoneoscopic nephrectomy. Another had a perivesical collection in the open nephrectomy group. No mortality related to the procedures occurred. CONCLUSION: Retroperitoneoscopic nephrectomy should be considered as the procedure of choice for dialysis dependent patients. This has all the benefits of minimally invasive surgery such as reduced blood loss, minimal post operative pain leading to faster convalescence. PMID- 19127796 TI - Antibiotic prophylaxis for orthognathic surgery: a prospective, comparative, randomized study between amoxicillin-clavulanic acid and penicillin. AB - OBJECTIVE: The present study was designed to evaluate the prophylactic efficacy after the correction of dentofacial deformities between short- and long-term penicillin and amoxicillin-clavulanic acid. MATERIAL AND METHOD: The present study was prospective, randomized, and double blinded. Patients were separated into four groups randomly: short-term and long-term (5 days) penicillin and short term and long-term (5 days) amoxicillin-clavulanic acid. RESULTS: One hundred twenty two patients were assigned randomly into the four groups. Infection developed in a patient in the short-term amoxicillin-clavulanic acid group and in a patient in the long-term penicillin group. CONCLUSION: There were no differences in infection between the two groups of antibiotics. Based on the present study, short-term penicillin is still the most appropriate choice for prophylactic antibiotic in orthognathic surgery. PMID- 19127797 TI - Idiopathic left ventricular tachycardia in children. AB - BACKGROUND: Idiopathic left ventricular tachycardia (ILVT) is a rare but well recognized clinical entity. The clinical characteristics and prognosis of this form of ventricular tachycardia (VT) in Thai children is not known. OBJECTIVE: To define clinical presentations, drug therapies, roles of radiofrequency (RF) catheter ablation, and the short-term outcome of these children in Thailand. PATIENTS AND METHOD: From April 1999 to June 2007, 10 patients were diagnosed as ILVT by specific electrocardiographic features and therapeutic response. All patients had a structurally normal heart. Data were collected retrospectively. Baseline clinical information, 12-lead electrocardiography (ECG) during VT, responses to drug therapy, results of RF catheter ablation therapy, and outcome were determined. RESULTS: Median age at presentation was 9.5 years (range, 3.8 to 14.0 years). Three patients (30%) were male. Eight patients (80%) were diagnosed as supraventricular tachycardia (SVT) before ILVT diagnosis. Median duration from SVT diagnosis to the correct diagnosis was 1.5 years (range, 0 to 6.0 years). Palpitation and chest pain were usual clinical manifestations while congestive heart failure was the presentation in one due to incessant tachycardia. Two patients had recurrent VT episodes during acute febrile illnesses. The majority of patients responded to intravenous verapamil. RF catheter ablation was performed in 3 patients with recurrence of the VT in one. CONCLUSION: Prompt recognition of the ILVTespecially in the emergency department is very important. Verapamil is effective for acute termination as well as prevention of VT recurrence. When VT is refractory to medical therapy, RF catheter ablation is safe and effective. The short-term prognosis was good. PMID- 19127798 TI - Sjogren-like syndrome after bone marrow transplantation. AB - OBJECTIVE: To study the incidence of dry eye in Sjogren-like syndrome, graft versus-host disease (GVHD) in hematological patients undergoing bone marrow transplantation (BMT). MATERIAL AND METHOD: Prospective, cross-sectional study in twenty-six patients that were planned for BMT (group I). Twenty-nine patients undergoing BMT before study were classified as group II no GVHD (9), and group III with GVHD (20). Thirty-two normal subjects were controls. All subjects were examined by slit lamp biomicroscopy and had their tear samples analyzed about tear osmolarity. They were also evaluated for aqueous tear production by phenol red thread test, Schirmer test without anesthesia, tear film stability by tear break-up time (TBUT), and rose bengal staining 2 weeks before BMT (for group I) as well as 6 weeks, 3 months, and 6 months after BMT. The patients with GVHD were followed up 1 month later. Main outcome measures were amount of tear production, tear film stability, and dry eye symptoms. RESULTS: Average aqueous tear production in group III was less than control and group II (p < 0.001). Mean TBUT in group III was faster than control (p < 0.001) and group I before BMT (p = 0.001). Mean score of rose bengal staining in group III was more than control and group I before BMT (p < 0.001). Keratoconjunctivitis sicca and red eye developed in 27.5%, and 20% of group III, with incidence of dry eye by Schirmer test without anesthesia (67.5%). This compares with group II having incidence of dry eye of 16.7%. However, 42.3% of group I before BMT had dry eye compared with 9.4% in the controls (p < 0.001). CONCLUSION: Trend of dry eye in patients with BMT and GVHD were higher than no-GVHD group. Doctors should be aware of ocular symptoms and signs of dry eye in patients with BMT and follow-up for proper management. PMID- 19127799 TI - Variation of hepatic artery by 3-D reconstruction MDCT scan of liver in Siriraj Hospital. AB - OBJECTIVE: To evaluate the prevalence of hepatic arterial variant in Siriraj Hospital. MATERIAL AND METHOD: A retrospective study that was approved by ethic committee of Siriraj Hospital. Between August and October 2006, 200 studies of abdominal MDCT were randomly sampled and multi-planar reformatted by using program Vitrea 2. Both axial, two- and three-dimensional images were evaluated for possible variants of hepatic vasculature. The results were classified by Michel classification and analyzed in percentage. Moreover, a small group (23 studies) that had digital subtraction angiography (DSA) was compared to know accuracy and Intercorelation between the two reviewers (Kappa value). RESULTS: Of 200 studies, 83.5% had type I, 1% type II, 6% type III, 0.5% type IV, 3.5% type V, 1% type VI, 0.5% type VIII, 1% type IX, 0.5% Type X, and 2.5% others type. Accuracy of MDCT for detection hepatic artery variation as compared with DSA was 78.3%. CONCLUSION: All variation is about 16%. Type III is most common variation of patients in Siriraj Hospital. PMID- 19127800 TI - Perception of medical personnel on informed consent for research participation in Phramongkutklao Hospital and Phramongkutklao College of Medicine. AB - OBJECTIVE: To evaluate the perception of medical personnel and medical cadets toward informed consent obtained from potential research participants. MATERIAL AND METHOD: The authors conducted a study using self-administered questionnaires which included questions about perception on informed consent regarding its objectives, investigator's role, vulnerable subjects, family involvement and children's assent. The answer for each question was graded into 5 scales. RESULTS: A selection of 380, 30.5%, 37.6% and 31.8% of 669 were attending staff residents, and medical cadets, respectively. A total of 85.5% agreed that informed consent in therapeutic trials should be obtained by their own doctors. A total of 75.3% agreed that the primary objective of informed consent was to protect investigators from lawsuits. A total of 60.8% agreed that participant spouses had to be involved in the informed consent process. A total of 79.5% agreed that permission from children was necessary in research conducted in children. CONCLUSION: The role of investigators in therapeutic clinical trial, primary objectives of informed consent, and role of spouse were misunderstood among medical personnel and medical cadets. Education on research ethics should concentrate on these issues. PMID- 19127801 TI - Bioequivalence study of the two 1.5 g cefoperazone and sulbactam IM injections in Thai healthy male volunteers. AB - OBJECTIVE: To perform a bioequivalence study of the two 1.5 g cefoperazone (1.0 g) and sulbactam (0.5 g) between Cefper and Sulperazon injections. MATERIAL AND METHOD: The present study was performed in 24 Thai healthy male volunteers who were intramuscularly injected a single dose of 1.5 g cefoperazone and sulbactam. A single dose, two periods, two sequences, double blind randomized crossover with a one-week washout period was used. Blood samples were collected before and at 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 4, 6, 8, and 12 hours after intramuscular injection and determined for cefoperazone and sulbactam plasma concentration by validated HPLC-UV methods. The pharmacokinetic parameters were analyzed by noncompartmental analysis and the ANOVA was carried out. RESULTS: Tax of both cefoperazone and sulbactam for volunteers who were injected with either Cefper or Sulperazon injection were not significantly different (p > 0.05). The 90% confidence intervals of the log of ratio of either C(max) or AUC(last) or AUC(inf) of both cefoperazone and sulbactam between 1.5 g Cefper and Sulperazon injections were within the bioequivalence range of 0.80-1.25. CONCLUSION: The 1.5 g cefoperazone and sulbactam injection of Cefper and Sulperazone used in the present study are bioequivalent. PMID- 19127802 TI - Angiomyofibroblastoma of the vagina. AB - In 2006, a 36-year-old Burmese woman presented with prolonged vaginal bleeding. She had undergone pelvic examination that disclosed pedunculated polypoid vaginal mass. It originated from the posterior vaginal wall. Local excision was done, resulting in good condition. Pathological diagnosis was angiomyofibroblastoma. Angiomyofibroblastoma is a rare, benign non-recurring lesion, mesenchymal tumor occurring mainly in the vulval region of premenopausal women and local excision with clear margins is an adequate treatment. PMID- 19127804 TI - Are children with myopia more intelligent? A literature review. AB - PURPOSE: Refractive errors are a serious worldwide problem. So far a few papers have described the relationship between refractive errors and intelligence. However, based on the growing interest into the relationship between refractive errors and intelligence quotient (IQ) we decided to present and discuss the latest results of the clinical studies on that subject. MATERIAL AND METHODS: A review of the literature concerning the relationship between refractive errors and IQ was done. RESULTS: In 1958 Nadell and Hirsch found that children in America with myopia have a higher IQ. A similar relationship has been described by other researchers from the USA, the Czech Republic, Denmark, Israel, New Zealand, and Singapore. In other related studies, it was reported that myopic children regardless of their IQ gain better school achievements--table 1. It was also observed that schoolchildren with hyperopia have a lower IQ and gain worse school achievements--table 2. Several hypotheses explaining the relationship between refractive errors and intelligence have been published. Recently, Saw et al. concluded that higher IQ may be associated with myopia, independent of books read per week, in schoolchildren. According to them "the association between genetically driven IQ and myopia of hereditary predisposition could be forged because of a pleiotropic relationship between IQ and myopia in which the same causal factor is reflected in both genetic traits. There may be similar genes affecting eye size or growth (associated with myopia) and neocortical size (possibly associated with IQ)". CONCLUSIONS: The conducted clinical observations suggest that children with myopia may have a higher IQ. This relationship is most probably determined by genetic and environmental factors. PMID- 19127805 TI - Prevalence of myopia and hyperopia among urban and rural schoolchildren in Poland. AB - INTRODUCTION: Myopia and hyperopia have a significant clinical meaning as they can be the cause of low visual acuity or even blindness. Therefore, there is a high demand for all clinical investigations regarding the development of the eye and the creation of refractive errors. Nevertheless, not many papers have been published around the world which compared the prevalence of myopia as well as hyperopia among metropolitan and provincial schoolchildren. Whereas, in Poland there was not a single paper yet published concerning this topic. That is why the aim of this paper is to describe the prevalence of myopia and hyperopia among urban and rural schoolchildren in Poland. MATERIAL AND METHODS: 2206 students were examined 1155 boys and 1051 girls, aged 10-14 years, mean age 11.9 (SD = 1.4). 614 boys in the age of 11.8 years (SD = 1.4) lived in the city, as 541 boys in the age of 11.9 years (SD = 1.4) lived in the countryside. 586 girls in the age of 11.8 years (SD = 1.5) lived in the city, as 465 girls in the age of 11.9 years (SD = 1.4) lived in the countryside. The examined students were Caucasian and lived in Szczecin, Poland or in villages located near Szczecin. The examination included retinoscopy under cycloplegia. The refractive error readings were expressed as spherical equivalent (SE). Myopia was defined as SE of at least -0.5 D, hyperopia as SE of at least +1.5 D. Astigmatism among students with myopia and hyperopia was smaller than 1 DC. Data analysis was performed using chi2 test. P values of less than 0.05 were considered statistically significant. RESULTS: It was observed that myopia occurred more frequently among children living in the city than in the countryside. 13.9% urban and 7.5% rural schoolchildren had myopia (p < 0.001)--table 1. Furthermore, it was found that hyperopia is less frequent among children living in the city than in the countryside. 7.1% urban and 30.8% rural students had hyperopia (p < 0.001)--table 2. It was determined that the average refractive error is lower among children from the city than the countryside--table 3. CONCLUSION: Living in an urban or a rural environment may have an influence on the occurrence of myopia and hyperopia among schoolchildren. PMID- 19127806 TI - [Evaluation of visual function among patients with exudative AMD treated with an intravitreal injection of triamcinolone acetonide]. AB - AIM: The aim of our study was to evaluate the visual acuity, visual field, retinal bioelectrical function and fluorescein angiography (FA) results of patients with exudative AMD treated with an intravitreal injection of triamcinolone acetonide (IVTA). MATERIAL AND METHODS: Visual acuity (Snellen chart), Humphrey automatic static perimetry (HASP) 30-2 W-W, mfERG were performed before, one month and three months after a single intravitreal injection of about 20 mg triamcinolone acetonide in 17 eyes of 17 patients (that did not meet the criteria for PDT). Fluorescein angiography was evaluated before and 3 months after treatment. RESULTS: There was no significant improvement in visual acuity after a month and three months after treatment when comparing to the initial examination. Although a slight improvement of 1-2 lines on Snellen chart was noted in individual cases. No significant changes were observed in static perimetry results after 3 months follow-up. In mfERG, there was a significant decrease in both, response density and P1 wave amplitude in fifth peripheral ring. In the rest of the rings, in comparison to the initial examination significant changes were not observed. In 76% of the analyzed eyes stabilization of CNV activity was noted in FA. In majority of eyes, the active CNV diameter did not change or was reduced and transformed into a scar. CONCLUSIONS: Three months after an intravitreal injection oftriamcinolone acetonide, no significant improvement of visual functions was noted in the observed group of patients with exudative AMD. A decrease of leakage in FA and a lack of a significant decrease in visual function may lead to a conclusion that IVTA treatment may slow down the natural course of the disease. PMID- 19127807 TI - [Estimation of treatment results and side-effects in patients with invasive uterine cervix cancer stage IIB-IVA treated with concurrent radiochemotherapy]. AB - PURPOSE: The aim of the study was a comparison between two treatment methods of uterine cervix cancer--concurrent radiochemoterapy (RT + CHTH) and radiotherapy (RT). The group of 106 patients, suffering from advanced uterine cervix cancer, was analysed. 52 patients underwent treatment with concomitant radiochemioterapy, while 54 patients only radiotherapy. The factors influencing treatment results were analysed. Analysis of overall survival, time to local relapse and time to distant metastasis were analysed. MATERIAL AND METHODS: An analysis was performed depending on deferent clinical parameters: stage, localization and diameter of the tumor, infiltration of vagina fornicis, depth and bilateral vs unilateral infiltration ofparametrials, anaemia, performance status and value of kerma. Occurence of early and late side-effects was analysed. RESULTS: Frequency of persisting tumor after treatment depended on stage, and kerma. It did not differ between groups. Analysis of overall survival, local relapse free-time and distant metastasis free-time showed no deference between groups. Radiochemotherapy gave better results in the group of patients without anemia. In multivariate analysis only stage showed to be an independent prognostic factor for risk of death, local relapse and distant metastasis. Side-effects (nausea, vomits, leucopenia, thrombocytopenia, early side effects from intestine and bladder; and late side effects from bladder) were observed more frequently in the group treated with radiochemotherapy; the difference was statistically significant. There was no difference between both groups in the frequency of recto-vaginal and vesicovaginal fistulas. CONCLUSIONS: 1. Radiochemotherapy compared with radiotherapy shows better results in patients with advanced uterine cervix cancer, if there is no anemia. 2. Radiochemotherapy increases the frequency of early and late side-effects, it could impact negatively the quality of life during and after treatment. 3. Stage of disease was the main prognostic factor in uterine cervix carcinoma, independently of the treatment scheme that was used. PMID- 19127808 TI - Impact of posttransplant diabetes mellitus on graft function in autosomal dominant polycystic kidney disease patients after kidney transplantation. AB - INTRODUCTION: Posttransplant diabetes mellitus (PTDM) remains the significant clinical problem and impairs the quality of life of renal transplant recipients. Negative influence of PTDM on graft function is associated with chronic allograft nephropathy, systemic infectious complications, recurrent infections or urinary tract infections. Some earlier studies suggest that patients with Autosomal Dominant Polycystic Kidney Disease (ADPKD) are at special risk of developing PTDM. The aim of our study was to assess the influence of PTDM on graft function in recipients with ADPKD and other causes of renal failure during 12-month follow up. Another aim of the study was to answer the question if the etiology of renal failure (ADPKD vs non-ADPKD) and the development of PTDM were independent predictors of graft function and if there was a significant interaction between them. MATERIAL AND METHODS: 98 pairs of kidney recipients from the same cadaveric donor were included into the study, with the ADPKD-related renal failure in one of the recipients, and different reasons for transplant in the other, with exclusion of the diabetic nephropathy. Diabetes was diagnosed according to American Diabetes Association (ADA) criteria. For statistical analysis the following parameters were included: age, sex, development ofposttransplant complications such as: PTDM, acute tubular necrosis (ATN) and early graft rejection, the presence of surgical and infectious complications (urinary tract infections excluded) and urinary tract infections as well as kidney function parameters such as serum urea, creatinine and uric acid concentrations 3, 6 and 12 months after transplantation. RESULTS: At the time of transplantation ADPKD patients were significantly older than non ADPKD patients (median 49.5 vs 45.3 years, p < 0.00008). There was no significant difference in incidence of PTDM between the study groups (19% in the ADPKD group and 18% in non-ADPKD group, p = 1.0). In ADPKD group no significant differences in serum urea, creatinine and uric acid following transplant were found between PTDM and non-PTDM subgroups. In non-ADPKD group, 3 and 6 months after transplantation, serum urea concentrations (77.5 mg/dL vs 54.0 mg/dL, p = 0.007 and 77.0 mg/dL vs 56.7 mg/dL, p = 0.016) and uric acid concentrations (7.7 mg/dL vs 6.8 mg/dL, p = 0.001 and 7.3 mg/dL vs 6.2 mg/dL, p = 0.034) were significantly higher in PTDM than in non-PTDM subgroup. Serum creatinine concentration was significantly higher in PTDM group 12 months following transplant (1.87 mg/dL vs 1.5 mg/dL, p = 0.035), with borderline significance 6 months following transplant (1.77 mg/dL vs 1.5 mg/dL, p = 0.057). In PTDM subgroup of ADPKD patients the percentage of infectious complications was slightly higher than in non-PTDM subgroup, but the difference was not statistically significant (37% vs 18%, p = 0.12), urinary tract infections were significantly more prevalent in PTDM subgroup (47% vs 18%, p = 0.013). In multivariate analysis no significant influence of ADPKD on any of the parameters of graft function was found. Significant interaction between ADPKD and PTDM presence for their influence on parameters of graft function was not found either what suggests that PTDM had a similar negative effect on these parameters in both ADPKD and non-ADPKD patients. CONCLUSIONS: 1. PTDM has negative influence on graft function independently of ADPKD diagnosis. 2. PTDM presence in ADPKD group significantly predisposes to urinary tract infections. PMID- 19127809 TI - Changes in cytokine concentrations in graft renal vein during reperfusion in patients with and without delayed graft function. AB - INTRODUCTION: The impairment of organ function derived from ischemia-reperfusion injury is still an important problem in solid organ transplantation. Cell alterations induced by ischemia prime the tissue for the subsequent damage that occurs during the reperfusion phase. Despite recent advances in immunosuppressive therapy, delayed graft function (DGF) remains an important problem after kidney transplantation. Different studies have related various clinical factors to DGF, such as donor age, recipient age, cold ischemia time, initial immunosuppressive regimens. The aim of present study was to examine the changes in cytokine concentrations in graft renal vein during the reperfusion in relation to the development of delayed graft function. MATERIAL AND METHODS: The study included 17 recipients of cadaveric renal grafts (10 males, 7 females, mean age 49 +/- 7 years, cold ischemia time 25 +/- 3 h)--8 with DGF and 9 without DGF. Levels of IL lbeta, IL-2, IL-4, IL-6, IL-8, IL-10, IFN-gamma, TNF-beta and TNF-alpha in renal graft vein plasma during 5 first min. of reperfusion were quantified by flow cytometry. RESULTS: The increased concentrations ofIL-6, TNF-alpha and IL-1beta were observed during reperfusion. However there were no statistically significant differences between patients with and without DGF. PMID- 19127810 TI - The effect of preservation solutions UW and EC on purine concentration in rat kidney. AB - INTRODUCTION: Ischemia/reperfusion injury in organ transplantation is a multifactor process that may lead to organ damage and primary graft dysfunction. Perfusion is a process which creates a possibility of graft injury. Preservation solutions are thought to diminish the ischemic injury and an appropriate choice of the solution should guarantee a better graft function and good prognosis for graft survival. The aim of the study was to examine the effect of preservation solutions University of Wisconsin (UW) and Euro-Collins (EC) on purine concentration in rat kidney. MATERIAL AND METHODS: The study was carried out on Wistar rat kidneys divided into 3 groups: kidneys perfused with 0.9% NaCl (control group), kidneys perfused with UW preservation solution, kidneys perfused with EC preservation solution. After bilateral nephrectomy the right kidney was immediately frozen in liquid nitrogen and the left kidney was placed in 4 degrees C UW, EC or 0.9% NaCl solution for 24 hours. The concentrations of purine nucleotides were determined using high-performance liquid chromatography (HPLC) method. RESULTS: The concentrations ofpurine nucleotides in renal tissue without cold ischemia did not differ significantly between rats perfused with 0.9% NaCl, UW and EC solutions. The tissue concentrations of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), total adenine nucleotides (TAN), guanosine monophosphate (GMP) and inosine monophosphate (IMP) were significantly increased, whereas the concentrations of adenosine (Ado), inosine (Ino), guanosine (Guo), hypoxanthine (Hyp) and xanthine (Xan) were significantly lower in rats perfused with EC solution in comparison to rats perfused with 0.9% NaCl. CONCLUSIONS: Purine concentration profile in rat kidney reflects a protective influence of EC and UW solutions on high-energy nucleotides in conditions when their depletion might be harmful to graft function. PMID- 19127811 TI - [HLA-G alleles and risk of early pregnancy loss]. AB - INTRODUCTION: To investigate whether HLA-G gene polymorphism is associated with adverse pregnancy outcome, we screened exons 2, 3, 4 of HLA-G gene in 59 women with reproductive failure and 36 fertile women. The HLA-G 10108 allele was more common in the group of examined women than in women from the control group (OR = 5,9; p = 0,09), but this difference was not statistically significant. RESULTS: We conclude that the HLA-G 10108 allele might influence pregnancy outcome in the Polish population, but further studies are needed in this regard. PMID- 19127812 TI - [Is prevalence of the "white coat effect" correlated with a high risk of late diabetic complications in type 2 diabetic patients?]. AB - BACKGROUND: On the ground of existing hitherto reports it is suggested that the "white coat effect" is not correlated with a high risk of cardiovascular diseases. Conducted studies did not ascertain a correlation between the value of the "white coat effect" and late diabetic complications. The aim of this study was to evaluate the risk of late diabetic complications: polyneuropathy, retinopathy and nephropathy in type 2 diabetic, hypertensive patients, in which the "white coat effect" was detected. MATERIAL AND METHODS: The study comprised 67 type 2 diabetic, hypertensive patients. In all cases blood count, creatinine and urea serum concentration, urine examination, kidney ultrasonography were performed as well as neurological and ophtalmological examinations. Blood pressure values were measured with the use of mercurial sphygmomanometer during clinic visits. In all patients ambulatory blood pressure measurements (ABPM) were conducted. The "white coat effect" was detected when mean "systolic office blood pressure" values were > 10 mmHg higher than mean systolic blood pressure in ABPM day and mean "diastolic office blood pressure" values were > 5 mmHg higher than mean diastolic blood pressure in ABPM-day. RESULTS: The patients were devided into two groups depending on the presence of the "white coat effect". Group 1 (n = 41) consisted of patients with the "white coat effect" and the second group (n = 26) without this phenomen. There were differences between the studied groups in systolic and diastolic blood pressure values estimated by traditional blood pressure measurement during clinic visits (p < 0.05). Systolic and diastolic blood pressure values estimated in ambulatory conditions with the use of ABPM were comparable in both groups (p > 0.05). The number of late diabetic complications in patients did not differ in both of the studied groups (p > 0.05). CONCLUSION: The "white coat effect" has not got an influence on polyneuropathy, retinopathy and nephropathy in type 2 diabetic patients with arterial hypertension. PMID- 19127814 TI - [Orthodontic evaluation of the results of surgical treatment of mandibular prognathism based on the cephalometric analysis using the Segner-Hasund method]. AB - PURPOSE: The aim of the study was the comparison of the cephalometric results occurring during orthodontic and surgical treatment of patients with mandibular prognathism. Results of patients treated surgically and of patients who underwent complex orthodontic and orthognathic treatment were compared. Additionally, the purpose of the research was to evaluate the psychical state of the prognathic patient: the affect the physical "difference" may have had on his life as well as his own opinion concerning the treatment and its outcome. MATERIAL AND METHOD: Cephalometric results using the Segner-Hasund method of two groups of patients were compared: group I (12 patients undergoing surgical treatment), group II (11 patients treated orthodonically and surgically). The X-rays in group I were taken just before the surgical treatment and at least 3 months after the treatment, and in group II, before orthodontic treatment and at least 3 months after surgery was preformed. Additionally patients were asked to answer a questionnaire concerning their self-esteem and approach to the treatment and its results. CONCLUSIONS: The comparison ofcephalometric results of the prognathic patient prior and post treatment gives a good insight on changes which take place. The study showed that better cephalometric results were obtained in group II i.e., in patients undergoing orthodontic--orthognathic treatment. Patients of both groups were pleased with the outcomes of the treatment (with a slight emphasis on group II), which lead to the conclusion that treatment of the prognathic patient has a positive influence on his self-esteem and way of life. PMID- 19127813 TI - Alveolar ridge sockets preservation with bone grafting--review. AB - INTRODUCTION: Alveolar bone seems to play a key role in providing support to the teeth, which are anchored to the bone by desmodontal fibers. The progressive alveolar bone resorption process occurs due to a loss of anatomic, biologic and mechanical factors. Mechanical stimulation of alveolar bone during mastication is crucial in keeping the teeth and underlying bone healthy. Tooth extraction leads to typical bone deficiency of ridge width and height of alveolar crest and reduces the possibility of placing screw titanium implants. When tooth extraction is necessary, trauma should be minimized during the procedure and bone preservation should receive careful attention. The literature has shown that early bone loss can be significantly reduced by socket grafting. The process of socket grafting requires an understanding of wound healing and an appreciation of the biological properties of the products available for socket grafting. Augmentative measures may, thus, be required to guarantee optimal prosthetic replacement of the lost tissue. Success or failure of augmentation procedures is dependent on revascularization and remodelling of the grafted bone into a vital, load bearing bone. In contrast to a visible three-dimensional change, the concept of remodelling refers to the internal turnover of bone, which is a coupled process where osteoclastic resorption and osteoblastic formation are more or less balanced. To restore alveolar bone loss and support efficient placement of dental implants, many different bone substitute such as autografts, allografts, xenografts, synthetic biomaterials and osteoactive agents have been proposed. In order to avoid harvesting an autograft, and thereby eliminating additional surgical procedures and risks, bone grafting materials and substitutes are alternative filler materials to be used for ridge augmentation. PURPOSE: To present a literature review about biomaterials applicable in alveolar ridge sockets preservation to future implants insertion. CONCLUSION: The maintenance of the dental alveolar bone after extraction depend on the attentive surgery procedure and the use of materials capable to maintain the prior space and be helpful in bone tissue healing. PMID- 19127815 TI - [Orthodontic effects of palatal suture expansion]. AB - INTRODUCTION: The aim of the work was the determination of the possibility of the palatal suture expansion with an orthodontic method depending on the age of patients, and the assessment of orthodontic treatment results. MATERIAL AND METHODS: The examination composed 40 individuals (24 female and 16 male) at the age of 10 and 6 months to 30 years and 1 month. Two groups were formed with regard to age. RESULTS: The palatal suture expansion was obtained in 18 individuals (that is 100% of the examined) of the first group at the ages of 10 years and 6 months to 15 years and 5 months and in 12 individuals at the ages of 16 years and 4 months to 30 years and 1 month in the second group, which amounted to 54%. In 10 persons there was no palatal suture expansion. CONCLUSIONS: The palatal suture expansion influenced in a statistically significant way an increase in the width of the palate, anterior and posterior width of the upper dental arch and a decrease in the index of the palatal height. The palatal suture expansion had no statistically significant influence on lowering the height of the palate and on the decrease in overbite, or changes in overjet. The age had no statistically significant influence on orthodontic changes. However, the age had a statistically significant influence on orthodontic changes assessed on the basis of the width of diastema. PMID- 19127816 TI - [Dentistry anomalies in patients with Lynch syndrome and familial adenomatous polyposis]. AB - PURPOSE: The aim of the study was evaluation of kind and frequency of dental anomalies in patients with familial adenomatous polyposis (FAP) and Lynch syndrome (HNPCC), and analysis of dento-osseus abnormalities on panoramic views. MATERIAL AND METHODS: FAP group consisted of 15 patients with mutation of APC gene, HNPCC group consisted of 30 patients with mutation of MSH2 or MLH1 genes. Control groups consisted of 15 and 30 patients matched by a ratio of 1:1 in relation with age and sex. Examination included: history, physical examination (extra- and intraoral) and assessment of panoramic view. DMF index, OHI-S index, clinical and radiologic PI index, occlusion and presence of pathologic dental abrasion and parafunctions were assessed in intraoral examination. RESULTS: The results are presented in figures and tables. CONCLUSIONS: 1. The kind and frequency of dental anomalies is comparable with changes typical to Polish population. 2. Osseus changes like osteomas, DBI's, hazy sclerosis and dental changes like supernumerary and impacted teeth are typical for patients with FAP. The frequency and appearance of this changes is higher than in general population. 3. Knowledge of kind and frequency of dental abnormalities may help in diagnosis and prevention of hereditary colon cancer by dentists. PMID- 19127817 TI - [Proceeding with patients having mental impairment and those affected by congenital development anomalies during orthodontic treatment]. AB - The aim of the work was to describe the specificity of proceeding with patients having mental impairment and those affected by congenital development anomalies during orthodontic treatment. At the same time, an attempt has been made at solving some problems concerning the treatment of patients with special needs. Based on available references three model syndromes of congenital development anomalies with coexisting mental impairment have been described: Down, Lejeune and Williams syndromes. It results from the descriptions and discussions included in this work that the possibilities of optimal cooperation and most effective treatment of patients affected by congenital birth defects and by mental impairment are conditioned, among other things, by: good contact between a doctor, a patient and a parent (guardian), availability and application of the best treatment solutions as well as coordinated implementation of a common therapeutic plan by members of an interdisciplinary team in specialized medical health centres. PMID- 19127818 TI - [The effect of environmental and genetic factors in the pathogenesis of periodontitis]. AB - INTRODUCTION: The pathogenesis ofperiodontitis has not been completely defined, therefore the study was aimed at evaluating combined effects of environmental factors as well as interleukin-1beta (IL-1beta) and FcgammaRIIa genotypes on the risk of periodontal disease. MATERIAL AND METHODS: The study was carried out on 52 patients diagnosed with periodontitis (P), 20 suffering from aggressive periodontitis (AgP), 32 with chronic periodontitis (CP) as well as 52 subjects with healthy periodontium (HP). Interleukin-1beta genotype was evaluated using PCR-RFLP, whereas FcdeltaRIIa polymorphism with the application of PCR-ASA method. RESULTS: The frequency of IL-1beta 1/1, 1/2, 2/2 genotype was as follows: AgP patients--75%, 25%, 0%, CP: 65.6%, 34.4%, 0%, and P: 69.2%, 30.7%, 0%, respectively, and did not differ significantly from HP. Alike, no significant differences were found in FcgammaRIIa genotype distribution. H/H, R/H, R/R genotypes were revealed in 40.0%, 35.0%, 25.0% for AgP, 43.8%, 40.6%, 15.6% in CP patients, 42.3%, 38.5%, 19.2% for P subjects as well as 36.5%, 48.1%, 15.4% in HP subjects. Multivariate analysis revealed a significant association between AgP and age, smoking and API > 50%. As for CP significant association was found solely for API > 50%. CONCLUSIONS: IL-1B(+3954) and FcgammaRIIa (CD32) gene polymorphisms are not associated with an increased risk of AgP and CP. PMID- 19127819 TI - [The reasons of tooth extraction taking into consideration the socio economical status during changes in the public health service in the West-Pomeranian voivodeship]. AB - INTRODUCTION: There are many possibilities of diagnosing and treating, but the fact of an increasing number of patients who have resigned from expensive procedures in place of teeth extraction is still noticeable. MATERIAL AND METHODS: Poll analysis based on the population of a big city--Szczecin and small towns in West Pomerania revealed that one's wealth considerably impacts both the frequency of visits in dental practices and the decisions regarding a mode of treatment. RESULTS: It further occurs that the most common reason of tooth extraction is caries and its later complications. Over 25% of patients from Szczecin and nearly 20% of interrogated people in provinces had to extract teeth because of an economical reason due to the lack of money for restoring treatment. It also appeared that a majority of people feel unsatisfied with the proceeding reorganization of the national health service and have not observed any positive tendencies effecting from this process so far. PMID- 19127820 TI - [Localisation of maxillary impacted canines by using a selected angle and linear indices]. AB - INTRODUCTION: The aim of the work was to present a diagnostic value of selected indices and angles in orthodontic-surgical treatment of impacted maxillary canine. MATERIAL AND METHODS: 134 records with panoramic views of patients treated in the Department of Orthodontics of the Pomeranian Medical University in Szczecin because of occurrence of impacted maxillary canine (IMC) constituted the study material. Both linear (for example: the relation of length to mesial-distal width of IMC, and relation of mesial-distal width of IMC to the mesial-distal width of medial incisor on the side of IMC) and angle indices (the angle between the long axis of IMC and occlusal plane) were calculated on the outline of each IMC and neighboring teeth. RESULTS: The study showed that values of described indices were dependent on the vestibular-palatal position of IMC. The relation of the length to the mesial-distal width of IMC assumed the highest values for vestibulary positioned canines with regard to decreased width of the crown of these canines on the panoramic view (a phenomenon of parallax). The relation of the mesial-distal width of IMC to the mesial-distal width of IMC of the opposite side in the group of persons with unilaterally IMC had lower mean values in case of vestibular IMC, identically like the relation of the mesial-distal width of IMC to the mesial-distal width of medial incisor on the side of IMC. The value of the relation of the distance of the root's apex of IMC from the median line to the distance of the incisor cusp of IMC from the median line amounted from 1.21 3.28, and its increase was evidence of a more verticular position of IMC. The mean value of the angle between the long axis of IMC and the occlusal plane (oz angle) oscillated between 51.6-85.6 degrees for unilateral IMC, and between 46.7 90 degrees for bilateral IMC, respectively. CONCLUSIONS: 1. The described indices can be supportive in the assessment of localizing unilateral and bilateral IMC. 2. They assume various values for right- and left-sided impacted upper canines, both palatally and vestibularly positioned. 3. The value of the oz angle is dependent on syntopie of IMC. PMID- 19127821 TI - [The application of the asymmetry index in assessment of mandible size in difficult diagnostic case]. AB - INTRODUCTION: Recognised and unrecognised bone diseases including maxilla and/or mandible may have influance on formation of malocclusions. In first stages of diseases the patients are directed or report for orthodontic treatment which starting need additional examinations mainly pantomographic views. In spite doing necessary additional examinations sometimes we can't recognise disorder like patient presented in our study. Then we can observate and if changes will begin disturbing the function--surgical intervention. AIM: Establishment of orthodontic treatment plan and explanation if during three years the dimension of asymmetry resulting from wrong growth right and left part of mandible and the estimation the rate of changes happening in this time. MATERIAL AND METHOD: In study we used the own asymmetry index to estimate the patient's pantomographic views who reported for orthodontic treatment because of occlusion disorders, facial asymmetry and discomfort of mastication and speech. The telerentgenographic lateral views in right and posterior-anterior (PA) projection were also done. We measured and estimated the own asymmetry index on pantomographic views. RESULTS: The radiographs reveal the asymmetry of left part of mandible. The comparative analysis of pantomographic views enables the estimation of changes happening in time and the telerentgenographic lateral views, PA and computer tomography (CT) confirm changes which increase the asymmetry. CONCLUSIONS: The asymmetry index is the instrument which enable the estimation of growth changes in mandible with unsteady aetiology and histopathological unrecognised, allow determine the growth rate and facilitate the permanent control the dimension of mandible asymmetry. PMID- 19127822 TI - Antibacterial activity of two Mineral Trioxide Aggregate materials in vitro evaluation. AB - INTRODUCTION: It has been documented that total removal of carious dentin, assessed according to clinical criteria, does not mean complete eradication of bacteria. For that reason in prevention of caries and pulp inflammations, application of the materials having high antibacterial properties seems to be indispensable. AIM: Determination of antibacterial activities of two white Mineral Trioxide Aggregate (MTA) preparations against bacteria of the genus Streptococcus. MATERIAL AND METHODS: Antibacterial activities of the materials were determined by diffusion method on a solid tryptic-soy medium (Tryptic Soy Agar, Oxoid). Wells were prepared on the medium with a puncher of 7 mm diameter and 5 mm depth (four for each material). The bottom was poured with 10 microlitres of liquid TSA for sealing. Bacterial suspension was inoculated with cotton swabs onto the medium surface and then the wells were filled with the materials prepared ex tempore. The plates were left in room temperature for 30 minutes and incubated at 37 degrees C for 24 h. Results were read by measuring the inhibition zones on day 1, 2, 3 and 8 of the culture. RESULTS: All the study samples demonstrated inhibition of bacteria of the genus Streptococcus. The antibacterial effect of MTA was maintained till the end of the experiment (day 8). ProRoot White showed slightly larger inhibition zone against Streptococcus salivarius and Streptococcus sanguis as compared to MTA white Angelus. The mean inhibition zones against Streptococcus mutans appeared the same for both test materials. CONCLUSION: The two MTA preparations assessed in the current study show good antibacterial action against the standard strains Streptococcus mutans, Streptococcus sanguis and Streptococcus salivarius. PMID- 19127823 TI - [Biofilm formation on root canal--review]. AB - It seems likely that one of the reasons for failures in the endodontic treatment is the presence of biofilm in root canals. Biofilm bacteria have a slower metabolism and higher resistance and virulence due to phenotypic changes. The occurrence of biofilms has been reported both inside the canal and on the external root surface. The results of many studies suggest that biofilm may be associated with refractory periapical periodontitis and is often caused by the coronal leakage. PMID- 19127825 TI - [Evaluation of attitude towards euthanasia expressed by first year medical students from Szczecin, Greifswald and Lund medical faculties]. AB - INTRODUCTION: Medical students' attitude towards euthanasia is a very important ethical problem because they may grapple with this question as future doctors. The aim of the study was to compare the attitude to euthanasia in the group of first year medical students from Pomeranian Medical University in Szczecin, Ernst Moritz-Arndt University Greifswald (Germany) and Lund University (Sweden). MATERIAL AND METHODS: The study is based on anonymous filling out of the questionnaire about euthanasia by first year medical students. 233 students (61%) answered the questionnaire. There were 65 Polish students, 71 German and 97 Swedish ones. In the group of respondents there were 129 (55%) women and 104 (45%) men. The average age was 22.3 years. RESULTS: 82% of questioned German students declared the acceptance of euthanasia and it was a significantly higher percentage than in comparison to 61% of Swedish students (p < 0.007) and 48% Polish ones (p < 0.0001). Poles were more often against euthanasia (29%) in comparison to 12% of Swedes (p < 0.02) and 3% of Germans (p < 0.001). Unnatural support of patient's life was the most often accepted by students clinical situation to use euthanasia. Significantly more Germans than Poles (79% vs 48%; p < 0.005) and Swedes (79% vs 50%; p < 0.02) accepted euthanasia in the group of questioned students declaring themselves as believers. CONCLUSIONS: German students in the highest percentage declared the acceptance of euthanasia and Polish ones--the highest objection. It may be connected with religious beliefs as the element of cultural differences among above three countries. It seems very proper to continue the study among older medical students. PMID- 19127824 TI - [Relationship of preoperative anxiety-state and anxiety-trait in patients qualified for coronary artery bypass graft surgery to the perception of postoperative pain and other pain complaints]. AB - INTRODUCTION: The study investigated relationships of postoperative pain following coronary artery bypass graft (CABG) surgery with two variables: (1) level of preoperative anxiety (anxiety-state) and (2) level of anxiety-trait. MATERIAL AND METHODS: Eighty three randomly selected male patients (44-66 years old), qualified for CABG surgery, participated in the study. All of them were patients at the Cardiosurgery Medical Centre of Pomeranian Medical University. It was a prospective clinical study. Anxiety-state and anxiety-trait were measured in the preoperative period using a Polish version of STAI. During the postoperative period the following factors of postoperative pain were measured, using a modified version of McGill Pain Questionnaire: average level of perceived postoperative pain, level of extreme pain, pain frequency, degree of pain regression following analgesia and number of other pain complaints. RESULTS: A significant relationship of the level of preoperative anxiety-state and anxiety trait with the degree of pain regression within the postoperative wound following the administration of medication (analgesia efficiency) was found (p < 0.01 and 0.001, respectively). Additionally, a significant relationship was found between the level of anxiety-trait with the level of perceived extreme postoperative pain (p < 0.01) and with the number of other pain complaints (p < 0.1). CONCLUSIONS: This investigation confirmed that relationships of anxiety and pain reported in other research are true also for CABG patients. It has been also observed that patients with a high level of preoperative anxiety-state and stronger anxiety trait respond worse to analgesic medication than patients with a low level of anxiety-state and anxiety-trait. Therefore, actions undertaken to reduce patients' anxiety may reduce patients' need of analgesic medications. PMID- 19127826 TI - The neurobiological interpretation of the mental functions in the work of Santiago Ramon y Cajal. AB - Cajal (1852-1934) first had contact with the fields of psychiatry and psychology through the experimental study of hypnotism. He interpreted this phenomenon as an aberrant failure of the machinery of the brain. It may be that Cajal's initial interest in the theory of hypnotic suggestion eventually led him to study the biological phenomena related to sleep and dreams, in the neurophysiological explanation of which he disagreed openly with Freud's postulates. Finally, Cajal studied the histophysiological mechanisms of the higher mentalfunctions (thought, intelligence, memory, perception, etc.). He proposed that the pyramidal cells of the cerebral cortex, which he called 'psychic cells', formed the substrate of these functions; he proposed the law of 'avalanche conduction', and he speculated on the possibility of a phenomenon of neuronal plasticity in relation to learning processes. After being awarded the Nobel Prize in 1906, Cajal's interests in psychological aspects began to decline. PMID- 19127827 TI - Hysteria and neurasthenia in pre-1914 British medical discourse and in histories of shell-shock. AB - Histories of shell-shock have argued that the diagnosis was subdivided into the categories hysteria and neurasthenia, and that the differential distribution and treatment of these diagnoses was shaped by class and gender expectations. These arguments depend on the presentation of hysteria and neurasthenia as opposed constructs in British medical discourse before 1914. An analysis of the framing of these diagnoses in British medical discourse c. 1910-1914 demonstrates that hysteria and neurasthenia, although undergoing redefinition in these years, were closely connected through the designation of both as functional diseases, and the role attributed to heredity in each. Before the war these diagnoses were perceived as indicators of national decline. Continuity, as well as change, is evident in medical responses to shell-shock. PMID- 19127828 TI - The Mental Treatment Act 1945 in Ireland: an historical enquiry. AB - The Mental Treatment Act 1945 had a decisive influence on the provision and development of psychiatric services in Ireland. This paper examines: (a) the historical and psychiatric backgrounds to the introduction of the Mental Treatment Act 1945; (b) the main provisions of the Act; and (c) the international context of the Act, especially in relation to institutional and legislative trends in other jurisdictions. PMID- 19127830 TI - Karl Ludwig Kahlbaum. 1899. PMID- 19127829 TI - Meseritz-Obrawalde: a 'wild euthanasia' hospital of Nazi Germany. AB - In 1939, Hitler authorized a programme of 'euthanasia' of children and adults with physical and psychiatric disorders. Initially, gas chambers were established at six psychiatric institutions in Germany and Austria. This programme was discontinued in August 1941 but the killings continued on an individual basis. Physicians selected patients who were unable to work or who required extensive care, and ordered the nurses to administer lethal doses of sedatives. Meseritz Obrawalde was a site for 10,000 of these killings. Using documents from the trial of one of Obrawalde's physicians, Hilde Wernicke, the era of 'wild euthanasia' is described and her rationale for participating in the killings is explored. PMID- 19127831 TI - Reading medicine in the Codex de la Cruz Badiano. PMID- 19127832 TI - The Jesus hermaphrodite: science and sex difference in premodern Europe. PMID- 19127833 TI - The mystical body of society: religion and association in nineteenth-century French political thought. PMID- 19127834 TI - "Science cannot stop with science" Maurice Blondel and the sciences. PMID- 19127835 TI - Between doctrine and clinical practice: nosography and semiology in the work of Jean-Etienne-Dominique Esquirol (1772-1840). AB - This paper makes a fresh analysis of the work of the French alienist J. E. D. Esquirol. In addition to a critical study of his classification of mental diseases, it analyses his main contributions to semiology: the difference between hallucinations and illusions, etc., and it also places the psychiatric clinic into the context of the so-called anatomoclinical mentality. PMID- 19127836 TI - Convergence and conflict: anthropology, psychiatry and feminism in the early writings of Madeleine Pelletier (1874-1939). AB - This paper reviews the early career and psychiatric writings of Madeleine Pelletier (1874-1939), between 1901 and 1906. Pelletier, a committed feminist and socialist, was the first woman to remove the barriers to women entering the French psychiatric profession, successfully passing the examination for the l'internat des hopitaux psychiatriques in 1903. Her involvement in La Societe d'Anthropologie de Paris and the Institut General de Psychologie provided a forum for her interest in psychology, both normal and abnormal. She turned increasingly to sociological explanations for mental illness, unwilling to accept the determinism inherent in degeneration theory. Although her career in psychiatry was cut short in 1906, her training and her debating skills, honed in the rationalism of the scientific and Free Masonic societies that she frequented, stood her in good stead in her subsequent career as a political militant. PMID- 19127837 TI - A Gentleman's mad-doctor in Georgian England: Edward Long Fox and Brislington House. AB - The late eighteenth and early nineteenth centuries were a period of particular innovation in the history of British psychiatry. Enlightenment ideas brought a change in attitudes to insanity, reflected in the growing prevalence of psychologically based treatment techniques being implemented in both public and private institutions. A new group of practitioners, specializing in the treatment and management of insanity, was emerging. One of the most prominent and successful was Dr. Edward Long Fox, a Bristol physician. His main venture was the establishment of Brislington House in 1806. Here he created a state-of-the-art asylum, catering mainly for the wealthier members of society. Its unique design, with seven distinct houses, enabled classification of patients according to social class as well as behavioural presentation. Within a context of safety and security, Fox sought to provide a therapeutic regime based on the principles and practices of moral management. PMID- 19127838 TI - Joseph Schorstein: R. D. Laing's 'rabbi'. AB - The Scottish psychiatrist R. D. Laing (1929-88) was one of the most influential but also one of the most controversial figures in twentieth-century psychiatry as he explored the boundaries in schizophrenia and serious mental illness in families. Joseph Schorstein (1909-76) was born in Moravia, graduated in medicine in Vienna, specialized in neurosurgery and eventually settled in Glasgow following wartime army service. Laing acknowledged the role of Schorstein as his 'spiritual mentor' and guide to the European intellectual world. This paper looks at the relationship between these two very different men and examines the career of Schorstein and the influence of his cultural background in Central Europe in shaping aspects of medical thought and culture in Glasgow. PMID- 19127839 TI - Julius Ludwig August Koch (1841-1908): Christian, philosopher and psychiatrist. AB - Being deeply rooted in a Christian faith and having much interest in moral and ethical issues, Koch published some philosophical works Die Wirklichkeit und ihre Erkenntnis (Reality and its knowledge; Koch, 1886), in which he tried to bring together critical Kantian philosophy and Christian conviction. In 1888 he published Kurzgefasster Leitfaden der Psychiatrie (Short Textbook of Psychiatry). The next work, entitled Die psychopathischen Minderwertigkeiten (Psychopathic Inferiority; Koch, 1891-93) became one of his fundamental texts on the concept of personality disorders, a concept still used today. Although psychiatric disorders are nowadays classified differently, Koch's work still has great importance for the concept of personality disorders; furthermore, he provides very exact and detailed descriptions of a multitude of psychopathological symptoms. PMID- 19127840 TI - The psychiatrist Auguste Forel and his attitude to eugenics. AB - Until the end of the 20th century Forel (1848-1931) was seen as an important neuroanatomist, a fighter against alcoholism, a researcher on ants and the author of Die sexuelle Frage. Forel's racist and eugenic views have been forgotten. Without losing sight of his merits, this article focuses on his attitude to eugenics, and will show that eugenic thinking--based on his main principles- permeated his work. PMID- 19127841 TI - How fury and lucid intervals may be proven. 1669. PMID- 19127842 TI - [Textual research on the motives of professionalization of the famous physicians in the past dynasties]. AB - According to the study on the biographies of 106 famous physicians in the past dynasties, their motives of professionalization are summed up and classified into two types, namely, the reasons of themselves and their families. Furthermore, two different types also could be summarized respectively as six and three reasons based on their own conditions. Meanwhile, it shows that many physicians were also influenced by social morals and cultural thoughts in their times. Finally, it concludes that their great interest, intensive motives, and determined attitude to traditional Chinese medicine (TCM) are very important to their fame and success, which also has practical significance to today's education of TCM. PMID- 19127843 TI - [Dried meats and the provisions of poisonous dried meat recorded in Er nian lu ling (Statutes of the second year) on Han bamboo slips unearthed in Zhangjiashan]. AB - It was a common social phenomenon to process, to store and to consume dried meat in the Qin and Han Dynasties. But some dried meat was poisonous, which did much harm to people's health and even influenced the social stability at that time. A provision of poisonous dried meat in Statutes of the Second Year is the earliest statutes in ancient China on preventing and dealing with food poisoning now discovered. For the purpose of maintaining the social stability in early Han Dynasty, they stipulated the principles and methods of disposing the dried meat, and established the penalty standard for the breach of the dried meat disposing regulations, which has great significance in medical history and legislative history. PMID- 19127844 TI - [The development of the cognition and treatment of apoplexy in traditional Chinese medicine]. AB - Surveying the developmental history of the cognition and treatment of apoplexy in traditional Chinese medicine, it could be divided into 3 phases, viz. the phase of "exogenous wind" before the Tang and Song dynasties, the phase of contention of "endogenous wind" during the Jin, Yuan and Ming dynasties, and the phase of compromising of traditional Chinese and consulting of western medicine of "equal importance of exogenous and endogenous wind" after the Qing dynasty. Through the development of these three phases, the cognition of cause of disease and pathogenesis of apoplexy was deepened continuously, and the method of treatment, prescription and materia medica were enriched further. Especially, with the introduction and usage of modern scientific technology, the diagnosis and treatment of apoplexy were more standardized, and the effect was improved constantly, reflecting the characteristic and superiority of traditional Chinese medicine. PMID- 19127845 TI - [A type analysis of the statutory infectious diseases in the Republican period]. AB - In the current academic circle, there are different opinions about the types of the statutory infectious diseases in the Republican period, including the nine-, ten-, eleven- and thirteen-type hypotheses being the most popular. They are different not only on the diseases types, but also about the time of its announcement. This article argues that there were eight kinds of statutory infectious diseases in 1916, i.e. cholera, dysentery, typhoid, smallpox, diphtheria, scarlatina,plague, and typhus; nine in 1928, with cerebrospinal meningitis being added; ten in 1944, with relapsing fever being added; after that, there were no changes. The appointed infectious diseases were initially relapsing fever and malaria; when the relapsing fever became the statutory one, then the appointed ones were the malaria and kala-azar. The establishment of the statutory and the appointed infectious diseases signified that the government of the Republican period had intervened in the administration of infectious diseases. PMID- 19127846 TI - [Brief introduction of the construction of legal system of traditional Chinese medicine in modern China]. AB - Since the foundation of PRC, the people's government attaches great importance to the development of traditional Chinese Medicine. With the efforts of people from Chinese medicine and the Party Central Committee's concern, the legal system of traditional Chinese medicine has been progressing step by step. Since 1949, the construction of legal system of traditional Chinese medicine can be divided into three periods with lessons and achievements, each with its own features. Through the retrospect of the history of our legal system of traditional Chinese medicine from 1949 to modern times, and its analytical summary, we could obtain a lot of inspiration for the construction of modern legal system of traditional Chinese medicine. PMID- 19127847 TI - [Research on psychiatry in the integration of Chinese and Western medicine in 1950's]. AB - In 1950's, the studies on psychiatry in the integration of Chinese and Western medicine was in its primary stage, which can be further divided into two periods, viz. the preparative stage, covering 1950 to 1954, during which doctors of traditional Chinese medicine were absorbed into the group of psychiatry research; and the preliminary development period, covering 1954 to 1958, during which there were more development of the clinical research on psychiatry in the integration of Chinese and Western medicine and in the Chinese herbal medicine, thus offering some effective new methods and drugs. PMID- 19127848 TI - [The establishment of Medical College in Philadelphia, the first medical college of North America]. AB - Modern medical education system of America was established in the 20th century. However, the beginning of American medical education was far early in the colonial times when the Medical College in Philadelphia was set up. The historical review on the establishment of the first medical college, Medical College in Philadelphia, is helpful to the understanding of the history of American medical education, the characteristics of modern medical education in the U. S., even to get some historical experience from the development of American medical education. PMID- 19127849 TI - [Textual research on circulated versions of Wen re lun (On epidemic warm diseases) and its related problems]. AB - The Wen re lun (On Epidemic Warm Diseases), written by Ye Tian-shi, a famous physician of epidemic warm diseases in the Qing dynasty, was originally a teaching record between the tutors and disciples. The book was originally anonymous, and there were two different circulated versions compiled by two different scholars. The Wen zheng lun zhi (Treatment of Warm Syndromes) was arranged by Tang Da-lie, and its first edition was Wu yi hui jiang (Collected Discourses of Physicians in Wu Region), an xylographic edition of Tang's Wenxin Thatched Cottage in Wu region block-printed in the 57th year of Qianlong (1792). The Wen re lun (On Epidemic Warm Diseases) was arranged by Hua Xiu-yun, and its first edition was Weisheng Tang edition probably in the 42nd year of Qianlong of the Qing dynasty (1777) with disordered book names, which should be unified and marked. Hua Xiu-yun wasn't Ye's follower, and he looked for and arranged Ye's medical cases because of the adoration to Ye Tian-shi. The texts of these two editions were the same, while the academic style differed substantially. PMID- 19127850 TI - [Brief research on the theory of cold and heat of the foods in Zhou li (The rites of Zhou)]. AB - The theory of cold and heat of the foods was recorded in the Zhou li * Tian guan * Shi yi (The Rites of Zhou * Ministry of State * Dietician), namely, "all the solid food should be as warm as the spring, all the soup should be as hot as the summer, all the catsup should be as cool as the autumn, all the drink should be as cold as the winter". The theory is one of the principles of dietetic therapy in accordance with the ancient viewpoint of "correspondence between human body and natural environment". It is significant for the life preservation of the people. However, this principle is not an absolute one. In the practice, warm or heat, cool or cold should be considered according to the individuals and the time. PMID- 19127851 TI - [Exploration on the fulminating plague in Bianjing in 1232 and the climatic factors]. AB - The plague occurred in Bianjing in 1232 was a serious event in the history of Chinese epidemic. It lasted for over 50 days, with a death toll of over 900,000. It is speculated that this is a mass epidemic of pneumonic plague and the pathogen was carried by the Mongolian Army when attacking Bianjing. At that time, the plague spread in Mongolian Army camp and finally involved the suburbs of Bianjing. After the army withdrawal, when Bianjing residents and soldiers went out of the city to collect foods and were unfortunately infected, and thus this dangerous infectious disease was transmitted into the city from the suburbs. Soon, the plague broke out and became epidemic in Bianjing during the 27-day period of 5 May-1 June, because of the cold snap rushed upon Bianjing on 1 June, the unusual weather was the motive for the mass epidemic of the plague. By then, it was popularly recognized as "exogenous cold damage". At present, facing the global climatologic changes, it is of great practical significance to explore the rich materials of plague and the climatic records accumulated in Chinese medicine. PMID- 19127852 TI - [The TCM physiological viewpoints in the Pre-Qin dynasty as interpreted from the medical texts from Mawangdui]. AB - There were great developments about the TCM since the last years of Warring States. The physiological viewpoints were focused on vessel (channel), zang-fu viscera and various structures of the body. Although there were divergences between medical texts and contents because of different viewpoints. It promoted medical experiences and knowledge fast and the characteristic faced more appears. Deep and extensive exploration about these physiological programs enabled relevant knowledge correlating one another, and the holistic body concepts of the Chinese medicine takes shape gradually. Because of various kinds of life-related knowledge gradually become enriched and complicated, the experiences turned into notions and theories, the focus of medical research was changed from "from outside to inside" to "from outside inside". At the same time, the originally exploratory process and the knowledge about the human body gradually tended to be the data--modular, systematical knowledge-network, and became more conveniently operated. PMID- 19127853 TI - [On contribution of Liu Wan-su's San xiao lun (treatise on three diabetic syndromes) to the diagnosis and treatment of diabetes]. AB - Liu Wan-su's San xiao lun is the earliest existing book solely dealing with the subject of diabetes in traditional Chinese medicine. Its main contribution to the diagnosis and treatment of diabetes is the thorough presentation of pathogenesis. It points out the cause of diabetes is the vacuity of kidney, spleen and stomach while the clinical manifestation shows repletion heat of heart. Its main emphasis is on the common pathogenesis of the three diabetic syndromes rather than their pattern differentiation. Thus, the treatment principle is enriching the yin of kidney and spleen and draining the heart fire with cold and damp drugs. The maneuver in the book is flexible and plays a pivotal role in the theoretical development of diabetes in TCM. PMID- 19127854 TI - [Important vaccine accidents in history of vaccine and their apocalypses]. AB - Although it has been widely acknowledged that glorious history of vaccination of past 200 years has been a successful feat in the protection of multiple disease calamities, contamination, incomplete attenuation or high virulence of the vaccine, laboratory regulation, injection and preservation problems have witnessed many accidents in the river of history. This review would summarize and comment on typical events of accidents regarding vaccines, within the framework designed to provide experience and apocalypses PMID- 19127855 TI - [The inspiration of the development of National Museum of Health and Medicine of USA]. AB - The National Museum of Health and Medicine, primary named Armed Medicine Museum, is subordinated to the Armed Forces Institute of Pathology, and is one of the few museums in the United States that collects archives and displays human remains. The museum was founded in 1862 to document the effects of war wounds and disease on the human body, the museum has a long history of collecting and preserving human remains from military and civilian sources. This collection provides a rich source of data for researchers in forensic medicine, anthropology, pathology and military medicine. The specimens comprising the anatomical collections are available for research, exhibition, and other educational purposes. The collections have attracted hundreds of domestic and foreign researchers. PMID- 19127856 TI - A virtuoso's history: antiquarianism and the transmission of knowledge in the alchemical studies of Elias Ashmole. PMID- 19127857 TI - Mandeville's ship: theistic design and philosophical history in Charles Darwin's vision of natural selection. PMID- 19127858 TI - The pointsman: Maxwell's demon, Victorian free will, and the boundaries of science. PMID- 19127859 TI - [Progress in the studies on small molecule IGF-1R inhibitors]. AB - The importance of insulin-like growth factor 1 receptor (IGF-1R) signaling in malignant behaviour of tumour cells is well established. Inhibiting the activity of IGF-1R may result in striking apoptosis in malignant cells growing. IGF-1R antibodies which are currently in phase I and II clinical trials and several IGF IR TKIs have preclinically been characterized. This review describes recent developments of small molecule tyrosine kinase inhibitors. PMID- 19127860 TI - [Progress in the research of carbon nanotubes as drug carriers]. AB - Research and development of new drug carriers are crucial to the research of drugs. Due to their unique hollow structure and nano-diameter, carbon nanotubes (CNTs) can be used as drug carriers. Functionalization of CNTs with peptides, proteins, nucleic acids or even drug molecules, the so obtained functionalized CNTs can be used as carriers to deliver bioactive molecules into cells without causing any toxicity. The research progress of CNTs as drug carriers in recent years is summarized, and the CNTs' cytotoxicity and their ability to penetrate cells are discussed, and the methods of functionalizing carbon nanotubes are also mentioned in the paper. Along with the advancement of CNTs in drug carriers system, the relationship between the way to functionalize CNTs and the so obtained modified CNTs' ability to penetrate into cells, including the effect of dimension, should be further studied. Preparation of functionalized CNTs with high solubility and low toxicity as drug carriers will be the main research areas in the near future. PMID- 19127861 TI - [Advances in the study of small peptides in targeted drug delivery system]. AB - Recently various peptide receptors which displayed the highest binding affinity and specificity with their peptide ligands by ligand-receptor have been exploited to develop drug delivery system which can directionally deliver drug to targeted cell. It is significant to study and applicate, including targeted drug delivery system mediated by bombesin receptor, somatostatin receptor, SynB3 receptor, LH RH receptor and other peptide receptor, et al. Several small peptide fragments were selected as carriers radicals combining doxorubicin, 2-pyrrolino-DOX, methotrexate, cis-platinum, and camptothecin to form hybrid cytotoxic analogs. These highly potent cytotoxic analogs have been designed as targeted anti-tumor agents for the treatment and study of various cancers that possess receptors for the carrier peptide. PMID- 19127862 TI - [Progress in the new nonnucleoside anti-HIV reverse transcriptase inhibitor-DCK]. AB - 3',4'-Di-O-(S)-comphanoyl-(+)-cis-khellactone (DCK) is a synthetic khellactone ester that exhibits potent anti-HIV activity with a mechanism distinct from clinically used anti-HIV agents. Several series of DCK analoges have been synthesized and evaluated for inhibitory effects against HIV. This review article describes recent progress in the discovery, structural modification, and structure-activity relationship studies of potent anti-HIV DCK derivatives. PMID- 19127863 TI - [Mechanism of apoptosis induced by SIRT1 deacetylase inhibitors in human breast cancer MCF-7 drug-resistant cells]. AB - The mechanism of apoptosis induced by SIRT1 deacetylase inhibitors in both human breast cancer MCF-7 and MCF-7 doxorubicin-resistant cells was studied. MTT assay was used to detect growth-inhibitory effect on the cells. Protein expression was detected by Western blotting. Chromatin condensation was detected by a fluorescent microscope after Hoechst 33342 staining. Cell cycle distribution was analyzed with flow cytometry. Apoptotic cells were detected with Annexin V staining. Nicotinamide (NAM) and Sirtinol, two SIRT1 deacetylase inhibitors, exhibited the similar growth-inhibitory effects on MCF-7/DOX cells and MCF-7 cells, but no potentiation of DOX activities. The arrest at G2/M phase was detected by flow cytometry in both MCF-7 and MCF-7/DOX cells after NAM treatment. Activation of caspase pathway in MCF-7 cells, such as the cleavages of PARP, caspase-6, -7, -9, were observed after exposure to NAM 50 mmol x L(-1), accompanied by the occurrence of chromatin condensation and Annexin V positive cells. However, the cleavages of PARP, caspase-6 and -7 in MCF-7/DOX cells delayed after exposure to NAM for 24 h and obviously increased at 48 h with appearance of chromatin condensation and Annexin V positive cells. SIRT1 deacetylase inhibitors show no cross resistance to MCF-7 drug-resistant cells, and the similar growth-inhibitory actions of them to MCF-7 sensitive and drug resistant cells by which it is mediated by activation of apoptotic caspase pathway. PMID- 19127864 TI - m-Nisoldipine attenuates monocrotaline-induced pulmonary hypertension by suppressing 5-HT/ERK MAPK pathway. AB - Effect of new calcium antagonist m-nisoldipine (m-Nis) on MCT-induced PH in rats and its mechanisms were investigated. Rats were injected with a single dose (60 mg x kg(-1)) of MCT subcutaneously to induce PH. Pulmonary haemodynamic measurement and lung tissue morphological investigations were undertaken. The MDA production and SOD activity in the serum were tested. PCNA, ERK1 and p-ERK expressions were analyzed by Western blotting. The expressions of 5-HT and PCNA were observed with immunohistochemistry. Results suggested that the PAP, right ventricular index and the degree of muscularization of small pulmonary artery were elevated markedly in MCT group, which was attenuated by m-Nis treatment. A significant reduction in MDA production and an increase in the SOD activity in the serum were also observed in all three m-Nis groups. The number of PCNA and 5 HT positive smooth muscle cells increased significantly in MCT group, and m-Nis treatment attenuated the expression obviously. Western blotting results suggested that the protein expression of PCNA and the ratio of p-ERK/ ERK1 increased markedly in MCT group and decreased by m-Nis. In conclusion, m-Nis protected against MCT-induced PH by decreasing PAP, right ventricular index, PAMSCs proliferation and pulmonary artery remodelling, which may be related to the reduction of 5-HT and the suppression of the ERK/MAPK signal pathway. PMID- 19127865 TI - Effects of red clover extract on the activation and proliferation of mouse T lymphocytes and the NO secretion of mouse macrophages. AB - The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide (NO) secretion of mouse macrophages stimulated by lipopolysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had little cell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators (NO, CD69, CD25, CD71), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3+ T lymphocytes. These data suggested that RCE might exhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages. PMID- 19127866 TI - [Effects of methopterin on osteoclasts and study of its mechanism of action]. AB - This study is to observe the effects of methopterin on the activation and bone resorption function of murine osteoclasts, which were obtained by induction from bone marrow cell and purified to the purity of 70%-80%. The mechanism underlying the inhibitory effects of methopterin on inflammatory bone destruction was explored. MTT method was used to determine the effect of methopterin on the proliferation of osteoclasts. Flow cytometric analysis was used to determine the effect of methopterin on the apoptosis of osteocalsts. TRAP stain, bone resorption lacuna stain and measurement of lacuna area were executed to determine the effects of methopterin on the activation and function of osteoclasts. ELISA method was used to determine the effect of methopterin on the MMP-9 secretion from osteoclasts. RT-PCR method was used to determine the effect of methopterin on the mRNA expression of RANK and MMP-9 in osteoclasts. The results showed that methopterin (0.1-10 micromol x L(-1)) inhibited the proliferation of osteoclasts, methopterin (0.1-10 micromol x L(-1)) could inhibit the activation and bone resorption function of osteoclasts and induced the apoptosis of osteoclasts. Methopterin (0.01-10 micromol x L(-1)) also decreased the mRNA expression of RANK, but only at 1-10 micromol x L(-1) decreased the mRNA expression of MMP-9. These results indicated that there were intense relation between the inhibitory effects on the activation and function of osteoclasts and the inhibition of inflammatory bone destruction by methopterin. PMID- 19127868 TI - [A new 8-O-4' neolignan from Glehnia littoralis]. AB - To study the chemical constituents from Glehnia littoralis, macroreticular resin column chromatography, repeated column chromatography on Sephadex LH-20 and reverse phase ODS were used to isolate the compounds whose structures were elucidated based on spectroscopic data (ESI-MS, 1D and 2DNMR). From EtOAc-soluble fraction of the ethanol extracts, a new 8-O-4' neolignan was isolated and named as glehlinoside D (1), along with five known compounds: (-)-secoisolariciresinol (2), ferulic acid (3), caffeic acid (4), vanillic acid (5) and syringin (6). Compound 1 is a new 8-O-4' neolignan. PMID- 19127867 TI - [A new flavonol glycoside from Baeckea Frutescens L]. AB - To study the chemical constituents of the traditional Chinese herb Baeckea Frutescens L., a new flavonol glycoside, named 6, 8-dimethylkaempferol-3-O-alpha L-rhamnoside (1), together with seven known compounds: quercetin (2), quercetin-3 O-alpha-L-rhamnoside (3), myricetin (4), myricetin-3-O-alpha-L-rhamnoside (5), gallic acid (6), ursolic acid (7) and 1,3-dihydroxy-2-(2'-methoxylpropionyl)-5 methoxy-6-methylbenzene (8) were isolated by using silica gel column chromatography, polyamide column chromatography and recrytallization. Their structures were identified on the basis of physicochemical properties and spectroscopic analysis. Among them, compounds 2-7 were isolated from this plant for the first time and compound 8 was first isolated from plant. PMID- 19127869 TI - Quantitative determination and pharmacokinetics of retinamido-ester in rat plasma by liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry. AB - A highly sensitive, rapid and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative determination of retinamido ester in rat plasma was developed and validated. A simplified protein precipitation with acetonitrile was employed for the sample preparation. The separation was carried out on an Agilent TC C18 column (150 mm x 4.6 mm ID, 5 microm particle size) with the mobile phase consisted of methanol-water-formic acid (93: 7: 0.1). Simvastatin was used as internal standard. The detection was performed on a trap-quadrupole tandem mass spectrometer by selected reaction monitoring (SRM) scan mode via atmospheric pressure chemical ionization (APCI). The range of calibration curve was 0.05-50 ng x mL(-1) and the limit of quantification was 10 pg x mL(-1). The intra- and inter-day precision values were between 95.97% and 104.43%, and RSD was between 4.63% and 10.69%, respectively. This method was applied to determine the pharmacokinetic parameters. The main pharmacokinetic parameters of retinamido-ester after oral administration via gastric gavage of 2.5, 5, 10 mg x kg(-1) were as follows, T(1/2): (11.28 +/- 7.23), (8.90 +/- 3.82), (8.01 +/- 5.65) h; AUC(0-infinity): (103.41 +/- 61.46), (190.23 +/- 74.99), (421.66 +/- 229.20) ng x h x mL(-1); MRT: (6.31 +/- 0.75), (5.98 +/- 0.71), (6.18 +/- 0.97) h; CL/F: (30.10 +/- 13.67), (29.58 +/- 10.59), (31.18 +/- 17.51) L x h(-1) x kg(-1); Vd/F: (414.94 +/- 159.82), (356.16 +/- 139.85), (369.28 +/- 322.72) L x kg(-1), respectively. PMID- 19127870 TI - [Determination of the fingerprint attribution ratio and process recovery of medicinal effectiveness components for TCM-compound prescription with quantified fingerprint method]. AB - By setting up the organic additive model of chemical fingerprints of TCM compound, the quantified fingerprint method had been established to solve the qualitative and quantitative analyses problems for both the fingerprint attribution ratio and process recovery of medicinal effective components in TCM compound prescription. The method firstly performs the qualitative analyses of the attribution ratios, and then the quantitative analyses, which can successfully disclose the results of attribution ratio and determine the process recovery of the medicinal effective components for TCM-compound prescription. Three optional methods were represented to assess the amount and distribution proportion of chemical compositions for single crude drug to compound prescription. In terms of components absorbed ultraviolet light, S5 (Radix Scutellariae) was assessed to be the most important crude drug containing much more effective components, and S7 (Radix Gentianae), S4 (Flos Lonicerae Japonica), S8 (Rhizome Anemarrhena) and S9 (Fructus Gardeniae) were second important crude drugs. The results showed lower process recovery of the medicinal effective components for eight batches of marketed preparations. Above all, the quantified fingerprint method can objectively and accurately reflect how high is the contribution of a single crude drug to the compound prescription, and quantitatively evaluate the process recovery of medicinal effectiveness components. PMID- 19127871 TI - [Determination of levosimendan and its main metabolites in human plasma with HPLC MS/MS method]. AB - This paper is aimed to develop rapid, sensitive and convenient HPLC-MS/MS methods for the quantification of levosimendan and its metabolites OR-1855 and OR-1896 in human plasma. According to the different natures of the compounds, two sets of liquid chromatography and ionization modes were used for determination the concentration of levosimendan and its metabolites OR-1855 and OR-1896 in human plasma, separately. Following protein precipitation with methanol, the levosimendan and internal standard (rosuvastatin) were separated on a Capcell MG III C18 column (35 mm x 2.0 mm ID, 3 microm) with the mobile phase consisted of methanol-15 mmol x L(-1) ammonium acetate-formic acid (55: 45: 0.02, v/v/v). A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated in the negative ion mode. Its metabolites OR-1855, OR 1896 and internal standard doxofylline were extracted from plasma by liquid liquid extraction with ethyl acetate. Chromatographic separation was performed on a Zorbax Extend C18 column (150 mm x 4.6 mm ID, 5 microm) with the mobile phase consisted of methanol-15 mmol x L(-1) ammonium acetate-formic acid (65 :35 :0.1, v/v/v). A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated at the positive ion mode. The linear concentration ranges of the calibration curves for levosimendan and OR-1855 and OR-1896 were 0.10-50.0 ng x mL(-1), 0.20-100 ng x mL(-1), 0.20-100 ng x mL(-1), respectively. The lower limits of quantification of levosimendan and OR-1855 and OR-1896 were 0.10 ng x mL(-1), 0.20 ng x mL(-1), 0.20 ng x mL(-1), respectively. The methods proved to be sensitive, simple and rapid, and suitable for the pharmacokinetic study of levosimendan injection. PMID- 19127872 TI - [Preparation of phycocyanin subunits liposomes and the photodynamic experiment on cancer cells]. AB - Phycocyanin subunits liposomes (PCS-lip) were prepared and its cellular uptake and photodynamic therapy (PDT) effect on cancer cells were studied. In the experiment, film dispersion method was used to prepare phycocyanin subunits liposomes; particle size and distribution were detected by zetasizer and transmission electric microscope; the effects of liposome as carrier on cell uptake in vitro were evaluated in S180 by using fluorescence microscope; and photodynamic therapy effect was assessed with MTT method. As shown in the results, the particle size mainly ranged from 80 nm to 160 nm, and average encapsulation rate was 42.3%. In the concentration of 100 microg x mL(-1), transfection rate reached (18.5 +/- 0.8)% at 2 h, (23.1 +/- 0.9)% at 4 h, keeping a balance in 5-6 h, and its photodynamic therapy effect in vitro improved with the increasing of concentration of phycocyanin subunits liposomes. In the concentration of 200 micro x mL(-1) cell survival rate of BGC-823 and S180 reached (45 +/- 5.2)% and (36 +/- 5.5)%, respectively, and the cell survival rate differentiation between PCS-PDT group and PCS-lip-PDT group reached 7%-11% (P < 0.05). In this study film dispersion method could keep the biological activity of phycocyanin subunits very well. Phycocyanin subunits liposomes will transfect cells more quickly than phycocyanin subunits in the same concentration, and in the same conditions, phycocyanin subunits liposomes have the better PDT effect on cancer cells as they were incubated with cells for 4 h. PMID- 19127873 TI - [Preparation of docetaxel-loaded pH-sensitive block copolymer micelles]. AB - Basing on the synthesis of pH-sensitive amphiphilic block copolymer poly (2-ethyl 2-oxazoline)-poly (D, L-lactide)(PEOz-PDLLA), this paper presents the preparation of docetaxel-loaded pH-sensitive block copolymer micelles using film dispersion method. The critical micelle concentration (CMC) was measured by pyrene fluorescent probe technique. The entrapment efficiency and drug-loaded amount were determined by HPLC. The morphology, diameter and surface potential of the micelles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential analyzer, respectively. The in vitro release behavior of DTX from polymeric micelles was investigated using dialysis method. The results indicated that the CMC, drug-loaded amount and entrapment efficiency of the micelles was 1.0 x 10(-3) g x L(-1), 15.0% and 91.1%, respectively. The micelles had a narrow size distribution, with a mean diameter of 28.7 nm. The micelle was globular-shaped and its zeta potential was (1.19 +/- 0.12) mV. In pH 7.4 PBS, docetaxel was released in a sustained manner from the micelles; while in PBS at pH 5.0, drug was released more rapidly, which suggested the pH-sensitive drug release behavior of the PEOz-PDLLA micelles. According to all the studies above, it can be concluded that the PEOz-PDLLA block copolymer micelles may be applied as promising drug delivery system for hydrophobic anti tumor drugs. PMID- 19127874 TI - Leaps of faith: is forgiveness a useful concept? AB - Using detailed clinical vignettes, the author argues that, despite the current idealization of the concept of forgiveness, the term has no place in psychoanalytic work, and there are some hazards to giving it one. Clinically, the concept of forgiveness is seductive, implying that there should be a common outcome to a variety of injuries, stemming fom different situations and calling for different solutions. Every instance of what we call forgiveness can be seen to serve a different defensive function. While the conscious experience of what is called forgiveness is sometimes confused with the unconscious process of reparation, the two can only be described at different levels of psychic life. Despite the fact that in 'the unconscious' there is no such thing as forgiveness, the term has an adhesive quality in our thinking that also blunts the analyst's appreciation of the aggressive components in the work. In a final vignette, the author illustrates an analytic outcome that has the appearance of forgiveness, but is best understood as the complex result of the everyday work of analysis. PMID- 19127875 TI - Off the beaten track: Freud, sound and music. Statement of a problem and some historico-critical notes. AB - The authors note that the element of sound and music has no place in the model of mental functioning bequeathed to us by Freud, which is dominated by the visual and the representational. They consider the reasons for this exclusion and its consequences, and ask whether the simple biographical explanation offered by Freud himself is acceptable. This contribution reconstructs the historical and cultural background to that exclusion, cites some relevant emblematic passages, and discusses Freud's position on music and on the aesthetic experience in general. Particular attention is devoted to the relationship between Freud and Lipps, which is important both for the originality of Lipps's thinking in the turn-of-the-century debate and for his ideas on the musical aspects of the foundations of psychic life, at which Freud 'stopped', as he himself wrote. Moreover, the shade of Lipps accompanied Freud throughout his scientific career from 1898 to 1938. Like all foundations, that of psychoanalysis was shaped by a system of inclusions and exclusions. The exclusion of the element of sound and music is understandable in view of the cultural background to the development of the concepts of the representational unconscious and infantile sexuality. While the consequences have been far reaching, the knowledge accumulated since that exclusion enables us to resume, albeit on a different basis, the composition of the 'unfinished symphony' of the relationship between psychoanalysis and music. PMID- 19127876 TI - Between private and public: towards a conception of the transitional subject. AB - Elaborating upon Winnicott's seminal contributions on the transitional object, the author proposes a conception of a transitional subject in which the patient comes into being simultaneously between private and public, subjective creation and material life, me and not-me. By anchoring subjective creation in the real world (including the body), the patient creates a basis for authentic psychesoma as well as for both personal and symbolic contributions to the world beyond omnipotence, including the world of other subjects. In this sense, intersubjective life is seen as predicated upon transitionality, with the patient seen as simultaneously coming into being as a distinctly personal subject and, in part, as a symbol. Clinical phenomenology is described and is interpreted with respect to the need within psychoanalysis itself for a third, and for a realm of meaning-creation that lies beyond privacy, omnipotence, and the dyad. PMID- 19127877 TI - Life and death in Freudian metapsychology: a reappraisal of the second instinctual dualism. AB - In this paper we re-examine the second instinctual dualism hypothesis introduced by Freud in Beyond the Pleasure Principle. We suggest that the life instinct hypothesis as something opposed to the death instinct does not seem to fit into this theory easily. On the other hand, death instinct turns out to be an internal necessity of Freudian metapsychological theory from the beginning of Freud's metapsychological writing. We shall argue, based on the ideas formulated in Beyond the Pleasure Principle and in later metapsychological texts, that Freud could not wholly justify the existence of an opposition and a symmetry between the two classes of instincts. Even though up to his last works Freud held on to this instinctual dualism, again and again his arguments lead to the idea that the life instincts should be regarded, ultimately, as death instincts. PMID- 19127878 TI - Leadership and regressive group processes: a pilot study. AB - Various perspectives on leadership within the psychoanalytic, organizational and sociobiological literature are reviewed, with particular attention to research studies in these areas. Hypotheses are offered about what makes an effective leader: her ability to structure tasks well in order to avoid destructive regressions, to make constructive use of the omnipresent regressive energies in group life, and to redirect regressions when they occur. Systematic qualitative observations of three videotaped sessions each from N = 18 medical staff work groups at an urban medical center are discussed, as is the utility of a scale, the Leadership and Group Regressions Scale (LGRS), that attempts to operationalize the hypotheses. Analyzing the tapes qualitatively, it was noteworthy that at times (in N = 6 groups), the nominal leader of the group did not prove to be the actual, working leader. Quantitatively, a significant correlation was seen between leaders' LGRS scores and the group's satisfactory completion of their quantitative goals (p = 0.007) and ability to sustain the goals (p = 0.04), when the score of the person who met criteria for group leadership was used. PMID- 19127879 TI - The handling of the transference in French psychoanalysis. PMID- 19127880 TI - Freud's concept of temporality: differences with current ideas. PMID- 19127881 TI - Panel report: hope in a time of cholera. Psychoanalytic reflections. PMID- 19127882 TI - On: The Donald Woods Winnicott Archives--preliminary announcement from the Winnicott Trust. PMID- 19127883 TI - Reality game: contemporary violence and denaturization of language. PMID- 19127884 TI - Public health facts--why don't they lead to healthy public policy? PMID- 19127885 TI - Breast cancer knowledge and attitudes toward mammography as predictors of breast cancer preventive behavior in Kazakh, Korean, and Russian women in Kazakhstan. AB - OBJECTIVES: To explore differences in breast cancer knowledge and attitudes toward mammography for women representing three ethnic groups (Kazakh, Korean, and Russian) and to determine how these factors affect breast cancer preventive practices in Almaty City, Kazakhstan. METHODS: A cross-sectional, descriptive study design was utilized. Face-to-face interviews were conducted with 500 women in Almaty City. A combination of descriptive (ANOVA) and multivariate analyses (structural equation modeling) was used to estimate differences in respondents' breast cancer knowledge, attitudes, and practices (KAP). RESULTS: Findings indicate that women may be influenced by their clinicians' advice to engage in breast cancer preventive practices. Multivariate models suggest that breast cancer knowledge and attitudes toward the effectiveness of mammography are significant determinants of breast cancer preventive practices among study participants. CONCLUSIONS: Clinicians should encourage women to engage in breast cancer preventive practices. Clinical and public health interventions should be aimed at both women and healthcare providers to use mammography as a tool for early detection of breast cancer in Almaty City, Kazakhstan. PMID- 19127886 TI - Behavioral and psychosocial influences of risk perception among Hungarian adolescents. AB - OBJECTIVE: The main goal of the present study was to assess adolescents' risk perception, namely, their estimates of the likelihood of smoking and alcohol related illnesses and accidents. METHODS: The survey was conducted among high school students in Szeged (N = 560, aged between 14-19 years; mean: 16.7; S.D.: 1.4 years). The self-administered questionnaire contained items on sociodemographics, risk perception, health risk behaviors, and certain psychosocial variables. RESULTS: Being a male was associated with risk perception only in terms of alcohol-related illnesses. Smokers, alcohol users, and those who engaged in reckless transportation, evaluated themselves as being at higher risk for substance related illnesses and accidents compared to those who did not report such behaviors. Those who tended to use a seat belt, however, estimated the same amount of risk for an accident as did those who tended not to use them. Social attitudes tended to positively, whereas personal attitudes tended to negatively influence perceptions of health risks. The role of social comparison and self-efficacy, however, depends on the nature of behavior. CONCLUSION: Besides behavioral influences, psychosocial variables also play a role in adolescents' risk perception. PMID- 19127887 TI - Analysis of parasuicide, psychiatric care and completed suicides, implications for intervention strategy (Czech Republic, 1996-2000). AB - BACKGROUND: This retrospective study analyses the differences between suicide with and without previous parasuicide. The Czech Republic was one of the countries with the traditionally highest level of suicide mortality. During collapse of the communist regime and deep societal changes in the Czech Republic after the year 1989 the escalation of suicides was expected. Mortality from suicides decreased, however the gender and age differences increased. METHODS: A total of 2,711 suicides in the Czech Republic (1996-2000) were studied. Effects of socioeconomic characteristics, psychiatric diagnosis and care, and lifetime history of parasuicide on the risk of death from suicide were estimated using logistic regression. RESULTS: Twenty-three percent of persons who committed suicide had a prior history of parasuicide and almost twenty percent of them received no psychiatric care after the attempt. Young males with basic education, economically active and diagnosed with substance abuse related disorders and the elderly were least likely to receive psychiatric care before their suicide death. CONCLUSIONS: Implications for age specific primary and secondary prevention are discussed. PMID- 19127888 TI - Restructuring public health in Slovenia between 1985 and 2006. AB - OBJECTIVE: This paper explores the developments in the public health infrastructure in Slovenia in the context of the sociopolitical and legislative changes in health care over the last 20 years. It assesses the responsiveness of the public health institutes in Slovenia to the various plans on public health developed by health policy makers over time METHODS: After an in-depth and externally validated search for key documents, we analysed the legislation, policy documents, research reports, theses, and other health policy papers related to the public health infrastructure in Slovenia. Findings were validated through consulting 3 external experts on public health in Slovenia. RESULTS: In the period discussed only few new services were added and health promotion was developed as an institutional field. Passivity in the past caused a lack of decisions on some traditional services in a changed economic environment. Moving from a passive supporter of the former infrastructure to an active promoter of the reform sets health policy as the main architect of the new public health building. CONCLUSION: Slovenia's "house" of public health was amended and refurbished, but a thorough reconstruction has not taken place. In order to face the future challenges in public health, the infrastructure will require increased efficiency, professional workforce development and better responsiveness. PMID- 19127889 TI - The average age of smoking onset in Germany--trends and correlates. AB - OBJECTIVES: Germany is rated among the countries with the highest prevalence of tobacco use in Europe. This paper analyzes whether the age of smoking onset has decreased in recent years. METHODS: Multivariable event data analyses were performed on the basis of the representative national cross-sectional study "Drug Affinity among Young People in the Federal Republic of Germany 2004". The survey involved a total net sample of 3032 individuals aged 12 to 25. RESULTS: Socioeconomic groups starting to smoke at significantly earlier age include those from the economically deprived areas in eastern Germany, low educational achievers and subjects in households with adult smokers. CONCLUSIONS: The average age of smoking onset has decreased further in the 1978-1992 birth cohorts. PMID- 19127890 TI - When to use the odds ratio or the relative risk? PMID- 19127891 TI - [Would you like to take some Plutonium 210?]. PMID- 19127892 TI - [ETS and respiratory health in adults: a real threat]. AB - Hazards due to active smoking are known for a long time. On the other hand, the importance of the harmful effects of passive smoking are less recognized, having been shown initially mainly in the pediatric population. However, the importance of significant increased risks on various respiratory pathologies of the adult are well known today, in a Swiss society where the number of active smokers remains high, leading to a high prevalence of passive smoking exposure On the basis of recent publications and updates of various organizations, we propose a review of the history, noxious mechanisms and effects on various respiratory pathologies of the exposure to passive smoke in adults. PMID- 19127893 TI - [The hyperventilation syndrome]. AB - The hyperventilation syndrome is a disease affecting children as well as adults. It predominates in female and may be debilitating. It is frequently associated with anxiety. The diagnosis, that is unfortunately often belated, is a diagnosis of exclusion and relies on the anamnesis, various non specific signs, on the Nijmegens score and on a hyperventilation provocation test. A specialized treatment allows, in most cases, a good control of ventilation and the disappearance of symptoms. PMID- 19127894 TI - [The solitary pulmonary nodule]. AB - The incidental discovery of a solitary pulmonary nodule while performing a CT scan of the chest is a very common clinical problem. The differential diagnosis is large but the main clinical challenge is to exclude or ascertain a neoplasia. The evaluation of preexisting risk factors and the analysis of morphological characteristics of the nodule allow the clinician to solve this challenge in a significant number of cases. When the nature of the lesion remains indeterminate a careful follow-up with volumetric determination is necessary for decision making. PMID- 19127895 TI - [Obesity and respiratory disorders]. AB - The prevalence of obesity has considerably increased during the past thirty years. Possible consequences of obesity on respiratory physiology include a restrictive disorder, changes in ventilatory mechanics and an alteration of respiratory drive. Apart from the well established relation between obesity and obstructive sleep apnea-hypopnea syndrome, obesity is associated with two other respiratory disorders. On one hand, epidemiological and animal data suggest a causal relationship between obesity and asthma. On the other hand, morbid obesity is associated, through an alteration of the respiratory drive involving leptin, with a diurnal and nocturnal alveolar hypoventilation defining the obesity hypoventilation syndrome. These data emphasize the necessity for the medical practitioner to investigate any respiratory symptomatology in obese patients. PMID- 19127896 TI - [Follow-up of patients with home mechanical ventilation: experience in Geneva, Switzerland]. AB - Home mechanical ventilation (HMV) is the treatment of choice for patients with chronic hypercapnic respiratory failure. Since the middle of the 1980's, the number of cases under HMV has markedly increased. Presently, at our centre (n=176 patients), obesity-hypoventilation syndrome (36%), and COPD (21%) are the most frequent indications for HMV. Classical indications such as sequellae of tuberculosis, or poliomyelitis, kyphoscoliosis, or neuro-muscular diseases, represent 25% only of our HMV population. Bi-level pressure support ventilators have replaced the volumetric ventilators used in the 80's in most situations. A specialized team of nurses monitors closely at home clinical evolution, and compliance, gathering an important amount of clinical data, and allowing the early detection of clinical deterioration. PMID- 19127897 TI - [Pneumocystis pneumonia in patients with immunosuppression other than HIV infection]. AB - Pneumocystis jirovecii is an opportunistic pathogen causing life-threatening pneumonia in immunosuppressed patients. The number of non-HIV immunosuppressed patients at risk for Pneumocystis pneumonia is rapidly growing. In contrast to HIV patients, there are no guidelines for Pneumocystis prophylaxis in other immunocompromised hosts. A detailed analysis of current literature data allowed us hereby to define the type of immunocompromised patients for whom evidence suggests a benefit for PCP prophylaxis. PMID- 19127898 TI - [When should we refer patients with cystic fibrosis to a lung transplantation program]. AB - Since most patients with cystic fibrosis (CF) die from respiratory failure, lung transplantation remains for many of them the ultimate treatment. The decision to put a patient on the transplantation waiting list is based on a list of criteria from international guidelines. We aimed to determine if a survival score could help determine when a CF patient has to be referred to a transplantation program. We applied this score to a small number of consecutive patients from our adult CF clinic, but found that it was of limited value for individual decision. Awareness of international guidelines and clinical judgement are essential to refer CF patients at proper time for lung transplantation. PMID- 19127899 TI - [What to do in case of disagreement between physician and patient: a few legal and ethical considerations]. AB - A patient may not demand an intervention that is not medically indicated. Such situations put the physician in a conflictual position that may jeopardize the physician-patient relationship. Exploring the different viewpoints may help explaining the sources of disagreement and reach a consensus. When this fails, the final decision in Switzerland belongs to the physician, including in case of an incompetent patient. A project of a revision of the Swiss civil code would confer decision-making responsibilities in such cases to the person closest to the incompetent patient. This is cause for concern given the poor ability of surrogates to accurately predict patient preferences and the additional emotional burden to surrogates. Early discussion with a competent patient and establishment of advance directives is therefore extremely important. PMID- 19127900 TI - [The consultation or behind the scenes...]. PMID- 19127901 TI - [From the mad cow to the scrapie of the European sheep]. PMID- 19127902 TI - [Benedict XVI, death and progress of science]. PMID- 19127903 TI - [Honour of French psychiatry]. PMID- 19127904 TI - [The gun at the temple of the Swiss health care]. PMID- 19127905 TI - Effectiveness of knowledge on family planning methods among pregnant mothers. PMID- 19127906 TI - Adherence is key to HIV treatment. PMID- 19127907 TI - Needs of family members of patients admitted in ICU. PMID- 19127908 TI - [Observation on the therapeutic effect of neck clustered needling on insomnia]. AB - OBJECTIVE: To compare therapeutic effects of neck clustered needling and medication on insomnia so as to search for a better method for treatment of insomnia. METHODS: Seventy-six insomnia patients were randomly allocated to an acupuncture group and a medication group, 38 cases in each group. The acupuncture group was treated by neck clustered needling, twice each week, 10 sessions constituting one course, and the medication group by oral administration of Alprazolam, before sleeping each day, 5 weeks constituting one course. The Pittsburgh Sleep Quality Index (PSQI) score was used for assessment of therapeutic effects, and the therapeutic effect and changes of PSQI scores before and after treatment were observed in the both groups. RESULTS: The total effective rate was 92.1% in the acupuncture group and 89.5% in the medication group with no statistically significant difference (P > 0.05). The cured rate of 34.2% and the cured and markedly effective rate of 84.3% in the acupuncture group were significantly higher than 7.900 and 36.9% in the medication group (both P < 0.01). Comparisons of the differences before and after treatment in scores of various factors between the two groups showed that the acupuncture group was better than the medication group in sleep quality, hypnotic use, activity of daily living and the improvement of the total score (P < 0.05 or P < 0.01), but the medication group was better than the acupuncture group in improvement of falling-asleep time (P < 0.05). CONCLUSION: The therapeutic effect of the neck clustered needling on insomnia is better than that of the medicine. PMID- 19127909 TI - [Effects of heavenly stem-prescription of point selection of needling methods of Ziwu Liuzhu on ischemic cerebrovascular diseases]. AB - OBJECTIVE: To observe effects of heavenly stem-prescription of point selection of the needling methods of midnight-noon ebb-flow on motorial and neurological functional deficit of ischemic cerebrovascular diseases. METHODS: The patients were randomly divided into a Ziwu Liuzhu group and a channel acupoint selection group. In the channel acupoint selection group, Jianyu (LI 15), Quchi (LI 11), Waiguan (TE 5), Zusanli (ST 36) were selected, combined with selected acupoints by syndrome differentiation. In the Ziwu Liuzhu group, on the basis of the channel acupoint selection group, according to the principle "the acupoints of the yang-channel were opened at yang-day-yang-hour and the acupoints of the yin channel were opened at yin-day-yin-hour", and acupoints were selected by the day hour acupoint-opening method of the Ziwu Liuzhu Najia method in Zhenjiu Daquan. The patients in the both groups began to be treated at the Chen hour (7:00-9:00) or Si hour (9:00-11:00) in the morning, 10 times constituting one course. They were treated for 3 courses. Improvement of the motor function symptom cumulative score, Fugl-Meyer (FMA) and improvement of neurological functional deficit were investigated. RESULTS: The FMA score (36.13 +/- 21.80) after treatment was significantly lower than (73.50 +/- 21.53) before treatment (P<0.01) in the Ziwu Liuzhu group, and (54.43 +/- 20.89) after treatment was significantly lower than (62.27 +/- 22.91) before treatment (P<0.05) in the channel acupoint selection group; the neurological functional deficit score (15.40 +/- 9.34) after treatment was significantly lower than (27.17 +/- 10.81) before treatment in the Ziwu Liuzhu group, and (23.97 +/- 1.30) was lower than (27.97 +/- 7.72) before treatment in the channel acupoint selection group. After treatment, FMA and the neurological functional scores significantly improved in the Ziwu Liuzhu group as compared with those in the channel acupoint selection group (P<0.05). CONCLUSION: Ziwu Liuzhu Najia point-selection method can effectively improve both nervous function and motor function in the patient of ischemic cerebrovascular disease. PMID- 19127910 TI - [Effects of acupuncture on nervous function in the patient of extensive cerebral infarction after operation]. AB - OBJECTIVE: To find a better therapy for extensive cerebral infarction. METHODS: Sixty cases with extensive cerebral infarction were treated by surgical decompression. After operation, they were randomly divided into an acupuncture plus medication group and a medication group, 30 cases in each group. The patients in the medication group were treated by routine western medicine, and the patients in the acupuncture plus medication group by the routine western medicine plus acupuncture 2 days after operation with Danzhong (CV 17), Zhongwan (CV 12), Qihai (CV 6), Zusanli (ST 36), Xuehai (SP 10) and Shuigou (GV 26) selected. They were treated for 10 days. The nervous functions were assessed by Chinese Stroke Nerve-function Rating Scale and criteria for clinical therapeutic effect. RESULTS: After surgical decompression, the death rate was 20.0%; after operation, acupuncture could significantly improve the neurological functional deficit degree of the patient. The total effective rate of 40.0% in the acupuncture plus medication group was better than 13.3% in the medication group (P<0.05). CONCLUSION: The surgical decompression is able to decrease the death rate of extensive cerebral infarction; after the operation, acupuncture and western medicine have cooperative effect on extensive cerebral infarction with a good therapeutic effect. PMID- 19127911 TI - [Combination of acupuncture with cupping increases life quality of patients of osteoporosis]. AB - OBJECTIVE: To observe therapeutic effect of "general regulation" acupuncture and cupping therapy on osteoporosis. METHODS: Sixty cases of primary osteoporosis were treated with the "general regulation" acupuncture-cupping therapy: warming needle moxibustion, plum-blossom needle, moving cupping and retaining the cup, thrice each week, for 3 months. The therapeutic effect was assessed by modified life quality scale for the patient of osteoporosis. RESULTS: Of the 60 cases, 3 cases did not complete the therapeutic course and 48 cases had a significant increase of life quality, the score of the life quality scale decreased from (67.45 +/- 15.67) before treatment to (42.28 +/- 27.89) after treatment with a significant difference (P<0.05). CONCLUSION: The "general regulation" acupuncture cupping therapy can effectively increase life quality of the patient with osteoporosis, which is an effective therapy for osteoporosis. PMID- 19127912 TI - [Observation on therapeutic effect of acupuncture at "cervical three points" combined with cake-separated moxibustion on cervical hyperosteogeny]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture at "cervical three points" combined with cake-separated moxibustion and routine acupuncture at Jiaji (EX-B 2) on cervical hyperosteogeny. METHODS: Single blind, randomized and controlled methods were used and 91 cases were divided into an observation group of 46 cases and a control group of 45 cases. The observation group was treated by acupuncture at "cervical three points" combined with cake-separated moxibustion, i.e. acupuncture at Chonggu (EX), Dazhui (GV 14), Taodao (GV 13) and moxibustion at pain points with cake made by Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba; the control group was treated with acupuncture at Jiaji (EX-B 2). Changes of sourness and pain of the neck, the shoulder and back, pressure pain and cervical activity scores were used for assessment of therapeutic effects. RESULTS: The markedly cured rate of 76.1% in the observation group was better than 42.2% in the control group (P<0.05). After treatment, the scores of all the items were significantly improved in the two groups (all P<0.05), and the observation group in sourness and pain of the neck, the shoulder and back, and cervical activity were better than the control group (all P<0.05). CONCLUSION: The therapeutic effect of acupuncture at "cervical three points" combined with cake-separated moxibustion on cervical hyperosteogeny is superior to routine acupuncture at Jiaji (EX-B 2). PMID- 19127913 TI - [Effects of acupuncture and moxibustion on blood vessels and the blood flow rate in patients with takayasu arteritis]. AB - OBJECTIVE: To observe the therapeutic effect of acupuncture and moxibustion on takayasu arteritis of brachiocephalic artery type and the mechanism. METHODS: Thirty-one cases were treated by acupuncture and moxibustion at main acupoint Renying (ST 9) and adjuvant acupoints selected according to the symptoms. The therapeutic effect and changes of color Doppler's ultrasonic flow image (CDFI) of relevant arteries in patients before and after treatment were investigated. RESULTS: The clinical total effective rate was 90.3%. Of the 31 cases, 5 cases were clinically cured, 16 cases were markedly effective, 7 cases improved and 3 cases were ineffective; and it was showed that acupuncture and moxibustion could enlarge the inner diameter of the injured blood vessel, increase the volume of blood flow and the elastic index of the injured blood vessel, and improve the abnormal velocity and the abnormal crest form of blood flow. CONCLUSION: Acupuncture and moxibustion has a good therapeutic effect on takayasu arteritis of brachiocephalic artery type; acupuncture and moxibustion can improve the stenotic degree of the injured blood vessel and abnormal hemodynamics of blood flow in the limb and brain. PMID- 19127914 TI - [Clinical observation on Tongdu Tiaojin needling method for treatment of blepharoptosis]. AB - OBJECTIVE: To compare therapeutic effects of Tongdu Tiaojin needling method and routine acupuncture method on blepharoptosis. METHODS: Fifty-eight cases of blepharoptosis were randomly divided into an observation group and a control group, 29 cases in each group. The observation group was treated by Tongdu Tiaojin needling method with Baihui (GV 20), Fengchi (GB 20), Houxi (SI 3), Yintang (EX-HN 3) selected, and the control group by routine acupuncture method with Cuanzhu (BL 2), Yangbai (GB 14), Sizhukong (TE 23) selected. They were treated for 3 courses and one course was constituted by 10 sessions. RESULTS: The cured rate of 41.4% and the effective rate of 82.8% in the observation group were better than 24.1% and 48.2% in the control group (P<0.05); after treatment, the palpebral fissure widened and the level of the palpebra superior shading the cornea was alleviated in the two groups, but the observation group was superior to the control group (P<0.05). CONCLUSION: Tongdu Tiaojin needling method has a definite therapeutic effect on blepharoptosis. PMID- 19127915 TI - [Clinical observation on acupuncture combined with diet control for treatment of simple obesity]. AB - OBJECTIVE: To search for an effective method for increasing the therapeutic effect on simple obesity. METHODS: Three hundred and thirty-four cases of simple obesity were randomly divided into an observation group (n=112), a control group I (n=112) and a control group II (n=110). The observation group was treated with acupuncture at Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), and diet control, the control group I with simple acupuncture and the control group II with diet control. RESULTS: Of the 334 cases enrolled, 66 withdrawn from the experiment (3 cases in the observation group, 5 cases in the control group I and 58 cases in the control group II ). The total effective rate was 91.8% in the observation group, 75.7% in the control group I and 73.1% in the control group I, with a significant differences as the observation group compared with the control group I or the control group II (both P<0.005); the effect-rebounding rate one year later in the observation group or the control group I was significantly different from that of the control group II (both P<0.001); and the number of cases withdrawning from the experiment in the control group II was significantly different from that in the observation group or in the control group I (both P<0.001). CONCLUSION: Acupuncture combined with diet control has a significant effect on simple obesity with a lower rate of the effect-rebounding, and it is easily accepted. PMID- 19127916 TI - [Treatment of hypomenorrhea by acupoint catgut embedding]. AB - OBJECTIVE: To observe therapeutic effect of acupoint catgut embedding on hypomenorrhea. METHODS: The patients were divided into an acupoint catgut embedding group and an electroacupuncture group. They were treated respectively by acupoint catgut embedding and electroacupuncture at Qihai (CV 6), Guanyuan (CV 4), Zigong (EX-CA 1), Ciliao (BL 32) , etc. The effects of the two therapies on the menses amount in different therapeutic periods were compared and their therapeutic effects were analyzed. RESULTS: The total effective rate was 100.0% in the acupoint catgut embedding group and 95.0% in the electroacupuncture group with no significant difference. One month and two months after treatment, the acupoint catgut embedding group in improving degree of the menses amount was better than the electroacupuncture group. CONCLUSION: Both acupoint catgut embedding and electroacupuncture can effectively treat hypomenorrhea, but the former more rapidly produces the effect and has a better short-term therapeutic effect. PMID- 19127917 TI - [Observation on therapeutic effect of intradermal needle combined with Tuina on lumbar disc herniation]. AB - OBJECTIVE: To observe the therapeutic effect of intradermal needle therapy combined with Tuina on lumbar disc herniation. METHODS: One hundred and fifty patients of lumbar disc herniation were treated with intradermal needle combined with Tuina. Trundle, rub the lumbar region for relaxing the lumbar muscles; pull and extend the lumbar region with traction and vibration, and pull the loin afterward and obliquely pull the loin, followed by compulsory straight-leg raising test, stretching sciatic nerve and hamstring muscle. Finally, rubbing method was used for relaxing the muscles of the lumbar and back regions. After Tuina therapy, intradermal needle embedding at local Ashi points, and Shenshu (BL 23), Yaoyangguan (GV 3), Zhibian (BL 54), Fengshi (GB 31), Yanglingquan (GB 34). The treatment was given once other day, 5 times constituting one course. After 2 courses therapeutic effects were assessed. RESULTS: One hundred and two cases were cured, 22 cases were markedly effective, 18 cases improved and 8 cases were ineffective, with a total effective rate of 94.7%. The symptom cumulative scores before and after treatment were (31.73 +/- 2.67) and (9.16 +/- 1.20), respectively, significantly decreasing (P<0.01). CONCLUSION: Intradermal needle combined with Tuina has a better clinical therapeutic effect on lumbar disc herniation. PMID- 19127919 TI - [Approach to the road of vigorously developing Chinese medicine by scientific location of channels and collaterals]. AB - Through researches of channels and collaterals and clinical practice of many years, the authors understand that Chinese medicine, which considers the human body as an interrelated, mutual constraints, whole, dynamic living system, has gradually become an important part of modern medicine. Channels and collaterals are a closed loop system which is communicated and linked by energy and information in the form of electromagnetic oscillation, reflecting many characteristics similar to quantum. Channels and collaterals are not a fixed organizational structure. Studies on channel and collaterals find that the track of the propagated sensation along channels (PSC) have the phenomenon drifting about. This exactly reflects the law of channels dynamically running. By information triggering and living resonance, channels and collaterals bring into play entirely regulative action. The innovative treatment of channels and collaterals followed by characteristics and laws of quantum can get a better curative effect. Theory of channels in the position of quantum information medical science provides an important breach for modernization of Chinese medicine. PMID- 19127918 TI - [Effects of warm needle moxibustion on bone mass density and biochemical indexes of bone metabolism in patients of postmenopausal osteoporosis]. AB - OBJECTIVE: To compare therapeutic effects of warm needle moxibustion and medication on osteoporosis and to study the mechanism. METHODS: Forty cases were randomly divided into an acupuncture group and a medication group, 20 cases in each group. The acupuncture group was treated by warm needle moxibustion at Dazhu (BL 11), Ganshu (BL 18), Shenshu (BL 23), Zusanli (ST 36), Yanglingquan (GB 34) etc. once other day, for 3 months; and the medication group was treated by oral administration of tablet Caltrate with Vit D2 for 3 months. The changes of bone mass density (BMD), estradiol (E2), osteocalcin (bone growth protein, BGP), urine calcium/creatinine (Ca/Cr) in the two groups before and after treatment and therapeutic effects were investigated. RESULTS: After treatment, BMD significantly increased (P<0.05, P<0.01) in the acupuncture group and did not signifi cantly changed in the medication group (P>0.05) with a significant difference between the two groups (P<0.05). After treatment E2 level significantly increased as compared with before treatment in both of groups (P<0.01); after treatment BGP significantly decreased as compared with before treatment in both of groups (P<0.01); after treatment Ca/Cr significantly decreased as compared with before treatment in the acupuncture group (P<0.05) ; af ter treatment, there were significant differences in BGP and Ca/Cr between the two groups (P<0.05 or P<0.01). The clinically controlled rate in the acupuncture group and in the medication group were 35.0%, 5.0%, respectively, the therapeutic effect of the acupuncture group being better than that of the medication group (P<0.01). CONCLUSION: The therapeutic effect of warm needle moxibustion on osteoporosis is better than that of oral administration of tablet Caltrate with Vit D2 and it can increase levels of hormones and delay bone loss. It is an effective method for preventing and treating postmenopausal osteoporosis. PMID- 19127920 TI - [Present situation and thinking of comparative studies on effects of acupoints and non-acupoints]. AB - The therapeutic effect of acupuncture is closely related to acupoints, and the specificity of acupoint indications is one of the key factors for regulatory and therapeutic effects of acupuncture. In recent years, a number of researches focusing on acupoint specificity have been done through comparison of the effects of acupoint and non acupoint. In this article, by analysis of recent 10 years' comparative studies of acupoints and non-acupoints, main problems in the existing researches are put forward including laying stress on the study of local effects, belittling the study on general effect; confused selection of acupoints and non acupoints; lack of high-quality multi-central, ran domized and controlled trials with large sample; and the train of thought of the future research are probed. PMID- 19127921 TI - [Mechanism of acupuncture anesthesia for analgesia of the operative region of thyroid gland]. AB - OBJECTIVE: To probe into peripheral nervous mechanisms of analgesic effect of electroacupuncture (EA) at Hegu (LI 4) and Neiguan (PC 6) on cervical region. METHODS: Twenty cases of thyroid disease who wished to receive acupuncture anesthesia, were stimulated with electroacupuncture at bilateral Hegu (LI 4), Neiguan (PC 6) in continued wave, intensity of 6-8 V, frequency of 10-20 Hz. After stimulation for 40 min, the operation was made. Before EA stimulation, 10 min, 20 min and 30 min after EA stimulation, changes of the physiologi cal functions including latent period, conduction velocity, wave peak amplitude of great auricular nerve were monitored and changes of pain sense were detected. RESULTS: After EA given at Hegu (LI 4) and Neiguan (PC 6) for 10 min, 20 min and 30 min, the pain sense significantly decreased (P<0.01), the latent period of great auricular nerve was shortened, and the conduction velocity was fastened and the wave peak amplitude raised with significant changes as compared with those before EA stimulation (P<0.01). CONCLUSION: EA at Hegu (LI 4) and Neiguan (PC 6) can produce a better analgesic effect on cervical region; EA at Hegu (LI 4) and Neiguan (PC 6) induces electro-physio logic changes of the nerve innervating the cervical region, showing increase of excitability; the nerves innervating Hegu (LI 4) and Neiguan (PC 6) and the nerves of cervical region possibly are homologous nerves. PMID- 19127922 TI - [Study on optimum stimulation parameter for Shuigou (GV 26) in Xingnao Kaiqiao needling method]. AB - OBJECTIVE: to investigate the influence of acupuncture parameter on acupuncture effect and to probe the optimum stimulation parameter for Shuigou (GV 26) in Xingnao Kaiqiao needling method. METHODS: The rat model of middle cerebral artery obstruction (MCAO) was replicated and "Shuigou" (GV 26) was stimulated by 9 acupuncture parameters combined by two factors at 3 levels, i. e. frequency 1, 2, 3 times/second, and needling duration 5 s, 60 s, 180 s. The effects of acupuncture at "Shuigou" (GV 26) with the 9 different acupuncture parameters on cerebral blood flow (CBF) in the MCAO rat were investigated by orthogonal test, factor analysis and one-way ANOVA. RESULTS: Acupuncture effect was influenced by acupuncture frequency, acupuncture duration and their combination. Among them, the acupuncture duration was the most important factor. The best acupuncture effect could be attained by using as long acupuncture duration as possibly with the fastest acupuncture frequency. CONCLUSION: In the Xingnao Kaiqiao needling method, acupuncture at "Shuigou" (GV 26) with fastest frequency and enough long duration can get the best acupuncture effect, which are the optimum stimulation parameters for "Shuigou" (GV 26). PMID- 19127923 TI - [Experimental study on changes of expressions of ER-alpha and ER-alpha mRNA in different hypothalamic nuclei and the effect of "preventive acupuncture and moxibustion" in ovariectomy rats]. AB - OBJECTIVE: To explore the mechanism of "preventive acupuncture and moxibustion" for regulating hypothalamic function in ovariectomy rats. METHODS: Thirty female SD rats, aged 3.5 months, were randomly divided into normal group, sham operation group, ovariectomy model group, preventive acupuncture group and preventive moxibustion group. After "preventive acupuncture and moxibustion" were previously given to the preventive acupuncture group and the preventive moxibustion group for one month, respectively. At the same time, ovarietomy was made in the preventive acupuncture group, the preventive moxibustion group and the ovariectomy model group. For the sham operation group, only a little fat around the ovary was removed without ovariectomy, and nothing was not applied to the normal group. Immunohistochemical and in situs hybridization methods were used to investigate changes of expressions of ER-alpha and ER-alpha mRNA in paraventricular nucleus, supraoptic nucleus and arcuate nucleus. RESULTS: In the model group the expressions of ER-alpha and ER-alpha mRNA in paraventricular nucleus and the expression of ER-alpha mRNA in supraoptic nucleus increased significantly (P<0.01), but the expression of ER-alpha dropped significantly in both supraoptic nucleus and arcuate nucleus (P<0.01). After preconditioning with acupuncture or moxibustion, the expression of ER-alpha mRNA in paraventricular nucleus significantly reduced (P<0.05), and the expression of ER-alpha had no significant change (P>0.05); the expression of ER-alpha in supraoptic nucleus and arcuate nucleus significantly increased (P<0.01 or P<0.05), and the expression of ER-alpha mRNA in paraventricular nucleus had no significant change (P>0.05). CONCLUSION: "Preventive acupuncture and moxibustion" at "Guanyuan" (CV 4) have an effect on the expressions of ER-alpha and ER-alpha mRNA in different hypothalamic nuclei in ovariectomy rats, which possibly is one of the ways for modulating the hypothalamic activity. PMID- 19127924 TI - [The importance of increasing students' subjective consciousness in teaching of the acupuncture and moxibustion sciences]. AB - Increasing the necessity and ways of students' subjective consciousness in teaching of The Acupuncture and Moxibustion Sciences are discussed. By cultivating students' learning interesting, the abilities of research, summarization and induction, and clinical practice as ways, teachers should make students establish subjective consciousness, which is the essential of reform and innovation of teaching method, and also is dual real interaction of "teaching" and "learning", so as to really reflect the idea of focusing on students themselves in teaching and the key of increasing teaching quality and efficacy of The Acupuncture and Moxibustion Sciences. PMID- 19127925 TI - [Gao Wu's studies and expounding about theories of acupuncture and moxibustion]. AB - The medical experts in the Ming dynasty paid attention to the study of acupuncture and moxibustion theories. The study methods and trait of thought, and expounding of relevant theories are worthy to be used for reference nowadays. In studies on acupuncture and moxibustion theories, GAO Wu took Neijing (Internal Classic), Nanjing (Medical Problems) as the criterion and set subject, classified and comprehensive analyze on classical theories, stressed origins and key points of the theories of channels and collaterals, acupoints, needling and moxibustion, etc. Particularly, in special subject studies and medical experts' academic systematization, both pictures and texts are combined, unite theories with clinical practice and others are worthy to be paid attention to and used for reference for modern acupuncture and moxibustion theory circle. PMID- 19127926 TI - Alpha Omegan presidential papers--June 2008. PMID- 19127927 TI - Dental ethics and professionalism. PMID- 19127928 TI - An overview of lasers in dentistry. PMID- 19127929 TI - Basic physics of laser interaction with vital tissue. AB - It is essential for any practitioner who uses lasers in their clinical practice to understand the basic physics of lasers. It is this knowledge that allows for an educated assessment of the clinical outcomes that lasers produce in our patients. It is also this understanding that provides a scientific basis for the visual feedback the clinician uses to vary parameters as needed to get the desired clinical results. It is the intent of this paper to discuss the very basic reasons why lasers affect tissues the way they do, and to synthesize the plethora of information dental practitioners are seeing regularly in dental journals. PMID- 19127930 TI - Restorative dentistry for children using a hard tissue laser. AB - This article will describe how a hard tissue laser is used in various restorative procedures. The erbium family of lasers (Er,Cr:YSGG and Er:YAG) are highly absorbed in water and, to a lesser extent, in tooth mineral. Because a carious lesion contains significantly more water than healthy enamel or dentin, the laser can offer some selectivity in removing the diseased material. Furthermore, these lasers can offer some degree of anesthesia. In these clinical photos, an Er,Cr:YSGG laser (Waterlase or Waterlase MD, 2780-nm; Biolase Technologies, Irvine, CA) was used. PMID- 19127931 TI - Lasers and soft tissue treatments for the pediatric dental patient. AB - Historically, oral soft tissue surgery on infants and young children was completed in the operating room under a general anesthetic agent. Many children were referred to an oral surgeon and required a physician's physical examination and medical clearance for a hospital admission. potentially placed a child at risk during the use of a general anesthetic for an elective procedure, which may not have insurance benefits. Traditional methods of oral surgery using scalpels or electrosurgery may produce significant postoperative discomfort and require sutures and prolonged healing. Lasers provide a simple and safe in-office alternative for children while at the same time reducing the chances of infection, swelling, discomfort, and scaring. PMID- 19127932 TI - The practical use of lasers in general practice. AB - The use of lasers in general dentistry is now an accepted--and to some extent, expected--treatment modality. Laser use can be either an adjunct to other procedures or the main form of treatment itself. For many procedures, lasers are now becoming the treatment of choice by both clinicians and patients, and in some cases, the standard of care. Though there are as many uses for lasers in general practice as the imagination can provide, this manuscript will address those indications for use that have received marketing clearance by the U.S. Food and Drug Administration (FDA). PMID- 19127933 TI - Sderot: dental medicine under fire. PMID- 19127934 TI - Suit for child's prenatal injuries not barred by limitations. PMID- 19127935 TI - Plaintiffs' expert's opinion rejected as 'conclusory'. PMID- 19127936 TI - OK: nurse terminated for medication error: court ordered unemployment benefits for RN. PMID- 19127937 TI - Nurse's professional liability insurance was not applicable. PMID- 19127938 TI - 401(k) plan asset allocation, account balances, and loan activity in 2007. AB - The bulk of 401(k) assets continued to be invested in stocks. On average, at year end 2007, about two-thirds of 401(k) participants' assets were invested in equity securities through equity funds, the equity portion of balanced funds, and company stock. About one-third was in fixed-income securities such as stable value investments and bond and money market funds. Although these relative shares have changed little over the past 12 years, the underlying fund composition has changed over time. About two-thirds of 401(k) plans included lifecycle funds in their investment lineup at year-end 2007. New analysis shows that at year-end 2007, more than 7 percent of the assets in the EBRI/ICI database were invested in lifecycle funds and one-quarter of 401(k) participants held lifecycle funds. Also known as "target date" funds, they are designed to simplify investing and automate account rebalancing. New employees continued to utilize balanced funds, including lifecyclefunds. Across all age groups, more new or recent hires invested their 401(k) assets in balanced funds, including lifecycle funds. At year-end 2007, 28 percent of the account balances of recently hired participants in their 20s were invested in balanced funds, compared with 24 percent in 2006, 19 percent in 2005, and about 7 percent in 1998. At year-end 2007, almost 19 percent of the account balances of recently hired participants in their 20s were invested in lifecycle funds compared with 16 percent at year-end 2006. 401(k) participants continued to seek diversification of their investments. The share of 401(k) accounts invested in company stock continued to shrink, falling by 0.5 percentage point (to 10.6 percent) in 2007. That continued a steady decline that started in 1999. Recently hired 401(k) participants contributed to this trend: they were less likely to hold employer stock. Participants' 401(k) loan activity was stable. In 2007, 18 percent of all 401(k) participants eligible for loans had a loan outstanding against their 401(k) account, the same percentage as at year end 2006. Most loans tended to be small, amounting to 12 percent of the remaining account balance, on average, similar to year-end 2006. At year-end 2007, the average account balance in the EBRI/ICI database was $65,454, compared with $61,346 at year-end 2006. 401(k) account balances varied with participant age, tenure, and salary. Individuals with account balances of less than $10,000 were primarily young workers or workers with short job tenures. In contrast, those with account balances in excess of $100,000 were primarily older workers or workers with longer job tenure. The year-end 2007 average account balance in the database was 6.7 percent higher than the year before, but does not accurately reflect the experience of typical 401(k) participants in 2007. To examine the experience of 401(k) participants, one must control for the impact of 401(k) plans or participants joining and leaving the database year to year. As with previous EBRI/ICI updates, analysis of a consistent sample of 401(k) participants (those that have been in the same plan since 1999) is planned; this additional analysis is expected to be published in early 2009. PMID- 19127939 TI - Improving drug treatment for children: are we making an impact? AB - At the beginning of 2009, we are on the threshold of mobilizing expertise in child health care and pediatric pharmacy and pharmacology in pursuit of worldwide major improvements in drug therapy for children. Momentum is built on progressive legislation on pediatric drug therapy from the US and Europe and on recent advances promoted by the WHO, including publication of an essential medicines list for children. Opportunities abound for research, educational initiatives, and practice improvements likely to bear early fruit in the form of better pharmacotherapy for children and youth. The most pressing challenge remaining is mobilization of a critical mass of caregivers, pharmacologists, pharmacists, and other child health professionals prepared to address their skills to this critically important task. PMID- 19127940 TI - The global alliance for pediatric pharmacology: the future is here and now. AB - The vast majority of drugs prescribed for children have not been approved by regulatory agencies for the pediatric age group and, in many cases, have not been studied appropriately. In July 2008, at a meeting of pediatric pharmacologists from over 30 countries and 5 continents in Toronto, ON, Canada, it became apparent that numerous pediatric drug studies are being conducted, often duplicating studies done elsewhere. This is also true for pharmacokinetic and pharmacodynamic studies, as well as for the development of pediatric formulations. Finding simple ways to inform the world about existing data may save time and facilitate efforts. PMID- 19127941 TI - Pediatric pharmacotherapy issues in Africa. AB - Pediatric pharmacotherapy in Africa is constrained by a variety of factors, not all of them unique to the continent. While affordability of medicines and the availability of sustainable financing for healthcare are constant challenges to African health systems, other issues of particular importance for pediatric care are also hampering access. These include the burden of disease in such settings, the lack of appropriate human resources for health, and the lack of child appropriate dosage forms, especially for ambulatory care. Access to medicines for children has been recognized as a global priority. Examples of developments that have the potential to improve pediatric pharmacotherapy in resource-constrained settings are the WHO Essential Medicines List for Children and the 'Make medicines child size' campaign. PMID- 19127942 TI - The European paediatric initiative: 1 year of experience. AB - The European Regulation on medicines for pediatric use entered into force on 26 January 2007. It changes dramatically the way medicines are developed for children. This regulation will increase availability of and information on pediatric medicines through high quality, ethical research. PMID- 19127943 TI - German Paediatric Research Network (PAED-Net). AB - Clinical trials in children are mandatory to generate data on new drugs as well as on drugs used off-label or for unlicensed indications. The EU Paediatric Regulation introduced in 2007 provides the background, goals, and requirements for pediatric clinical trials. The German Paediatric Network (PAED-Net) was established in 2002 with a public grant from the German Ministry of Education and Research with the aim of developing competence, infrastructure, networking, and education for pediatric clinical trials. The network, consisting of six pediatric clinical units and a coordinating office, has progressed very well and has achieved valuable improvements concerning the conduct of pediatric clinical trials. In addition, training programs have encouraged knowledge about clinical trials in investigators and nurses and have made medical professionals as well as the public aware of the need and advantages of trials in children. Although these trials are still challenging from the ethical, scientific, and clinical points of view, quality-controlled data are the prerequisite for sharing therapeutic innovations for children and for applying the principles of evidence-based medicine to the youngest patient population. PMID- 19127944 TI - The National Institute for Health Research Medicines for Children Research Network. AB - The UK has established the National Institute for Health Research Medicines for Children Research Network to provide infrastructure to facilitate the conduct of randomized trials and other well designed studies sponsored by the pharmaceutical industry and public funding bodies. Clinical Studies Groups of experts establish priorities and assist in the development of protocols whilst Local Research Networks and an Extended Neonatal Network facilitate their implementation and conduct. Two Clinical Trials Units and a co-ordinating center assist in the development and conduct of studies and performance manage the network. Work streams on formulation and experimental medicine have been established. As of August 2008, 16 commercially sponsored studies had been adopted (from 12 different companies) and 65 publicly sponsored studies. PMID- 19127945 TI - Drug therapy for children in China. AB - The population of China is huge and includes 400 million children. For China, as for other countries in the world, there remain a number of questions from the past about drug therapy for children, such as unlicensed and off-label drug usage. Recently, China has updated Good Clinical Practice (GCP) guidelines, including those for pediatric drug clinical trials. Pediatric pharmacology research units need to be set up throughout the country. To provide better medicines to children in China is an enormous undertaking. PMID- 19127946 TI - The Task-force in Europe for Drug Development for the Young (TEDDY) Network of Excellence. AB - The Task-force in Europe for Drug Development for the Young (TEDDY) was established in 2005 to contribute to the promotion of safe and efficacious medicines for children in the context of the impending European Paediatric Regulation that finally came into force in January 2007. The project includes seven objectives and 12 Work-Packages encompassing the main aspects of the development and use of pediatric drugs. TEDDY represents a new entity in the pediatric pharmaceutical field, differing from a Scientific Society, a network for developing research or trials, or a consultative regulatory body. The ambition of TEDDY is to support the existing pediatric networks, societies, and regulatory bodies in performing innovative initiatives, including those in areas in which such undertakings would not be feasible without supportive action. To accomplish its aim, TEDDY has focused on three different actions: (i) increasing awareness about the Paediatric Regulation revolution; (ii) reaching consensus on terms and instruments to be used for common research; and (iii) favoring close relationships among different stakeholders and partners from different EU Member States. After 3 years of activities, many results have been produced by the Network: surveys, databases, expert opinions, and recommendations. Linking together different stakeholders, including industry and patient associations, as well as academia and research centers, the Network has contributed to increasing awareness and participation in the Paediatric Regulation. In addition, many papers detailing original results have either been published or submitted for publication in peer-reviewed journals. TEDDY is an original Network whose identity and role as a catalyzer of initiatives related to the use and development of pediatric drugs needs to be better clarified in the near future. Of particular importance is the need to reach consensus on best practices. The lack of a common view on pediatric research requirements among stakeholders across Member States remains the main challenge to be overcome. PMID- 19127947 TI - Both parental psychopathology and prenatal maternal alcohol dependency can predict the behavioral phenotype in children. AB - OBJECTIVE: To identify whether a child's behavior phenotype can be predicted by parental psychopathology and/or prenatal maternal alcohol dependency by using the Child Behavior List (CBCL) as a screening tool. METHODS: A retrospective cohort of four non-exclusive groups of children (aged 8-15 years) was studied: (i) children exposed to alcohol in utero (n = 25); (ii) children not exposed to alcohol in utero (n = 46); (iii) children exposed to parental psychopathology (n = 37); (iv) children not exposed to parental psychopathology (n = 34). To distinguish between the effects of alcohol and parental psychopathology, the children were further subdivided into groups with alcohol exposure in utero and parental psychopathology (n = 23), and psychopathology without alchohol exposure (n = 14). Each child was assessed with the CBCL. Subscale scores and selected subscale items were compared between the groups using t-tests and regression analysis. RESULTS: Children exposed to alcohol in utero scored significantly lower than unexposed children on school competency (p = 0.015). They were more likely to attend special classes (p = 0.048), repeat a grade (p = 0.011), and display more disobedience (p = 0.039) and vandalism (p = 0.033). For special classes and disobedience at school, gender proved to be a significant predictor, while maternal alcohol dependency was a significant predictor of vandalism and repeated grades. Children with parental psychopathology differed from children without parental psychopathology in the anxious/depressed (p = 0.04), social problems (p = 0.004), and attention problems (p = 0.04) subscales. The subscale items that were significantly different between the groups were nervousness (p = 0.002), self-consciousness (p = 0.019), feelings of worthlessness (p = 0.041), loneliness (p = 0.005), and difficulty with concentration (p = 0.02). Parental psychopathology was a significant predictor of all five items. Age and gender, however, were significant predictors only of difficulty with concentration. No significant differences were found when the groups with alcohol exposure in utero and parental psychopathology, and psychopathology without alcohol exposure were compared. In summary, parental psychopathology was a significant predictor of a child's internalizing behavior, as well as social problems, whereas alcohol exposure was more predictive of externalizing behaviour. CONCLUSION: Parental psychopathology and prenatal exposure to maternal alcohol can contribute to the child's behavioral phenotype as measured by the CBCL. Therefore, the CBCL can be used to screen for such behaviors. PMID- 19127948 TI - Potential role of the placenta in fetal alcohol spectrum disorder. AB - BACKGROUND: Mediators of susceptibility to alcohol-related toxicity in the prenatal environment are relatively unknown. The placenta has been proposed as a potential source of variability in the fetal environment, primarily through its significant metabolic capabilities. Meconium is a toxicological matrix unique to the developing fetus that offers an opportunity to quantify fetal exposure to alcohol through the analysis of fatty acid ethyl esters (FAEE) [nonoxidative ethanol metabolites]. OBJECTIVE: To explore the role of the placenta as a potential source of variability in mediating fetal alcohol exposure via meconium analysis of FAEE, by combining a clinical case report involving human dizygotic twins and a series of guinea pig littermates. METHODS: Meconium was collected from a pair of dizygotic twins with clinical suspicion of prenatal alcohol exposure and analyzed for FAEE to confirm prenatal alcohol exposure. Meconium was also collected from a series of guinea pig pups prenatally exposed to alcohol at a daily dose of 4 g/kg/day administered to the pregnant dam. FAEE levels were analyzed by gas chromatography with flame ionization detection following liquid liquid extraction from meconium. RESULTS: The dizygotic twin pair yielded positive meconium FAEE results (>2.00 nmoL/g) in the female twin (2.21 nmoL/g) and no detectable FAEE in the male twin. A total of 15 meconium samples were collected from 15 pups of five pregnant guinea pig litters. With the exception of one pair of littermates, meconium FAEE concentrations differed substantially within each litter (FAEE expressed in units of nmoL/g): litter 1 (0.996, 4.43, 1.36); litter 2 (5.17, 4.15, 0.00); litter 3 (5.16, 5.27); litter 4 (18.57, 8.26, 7.46); litter 5 (0.00, 4.32, 0.00, 1.27). CONCLUSIONS: Identical maternal ethanol exposure levels produced differing levels of fetal exposure in a dizygotic human twin pair and a series of guinea pig littermates as evidenced through FAEE meconium analysis. These data indicate that the placenta may have a previously unappreciated role in mediating ethanol-induced fetal injury. PMID- 19127949 TI - Passive environmental exposure to cocaine in Canadian children. AB - BACKGROUND: Hair testing is commonly used to confirm potential drug exposure in children living with drug users, as well as abstinence in their caregivers. OBJECTIVE: To examine differences across pediatric age groups in the relationship between caregiver use of cocaine and cocaine exposure in children. METHODS: We determined concentrations of cocaine and its metabolite, benzoylecgonine, in the hair of 19 child-caregiver pairs to estimate the pattern of exposure of the children according to age; concentrations in the caregivers' hair were used as a surrogate marker for the intensity of environmental exposure. Cocaine and benzoylecgonine concentrations in hair were determined by immunoassay. RESULTS: A significant correlation was observed between cocaine concentrations in the hair of infants and their caregivers (Spearman rho = 0.87; p = 0.005; n = 8), and the absence of a correlation in older children. These results suggest that environmental exposure plays an important role in the accumulation of cocaine in the hair of infants. CONCLUSION: Measurement of cocaine hair concentrations can allow estimation of the degree of environmental drug exposure in young children. Infants seem to have a disproportionately increased risk for systemic exposure, compared with older children. PMID- 19127951 TI - Australian report on pediatric medication issues: is any magic happening in the 'Land of Oz' to save the therapeutic orphan? AB - Australian prescribers have faced many of the same issues as those in other developed countries in relation to medicines for children. Australia represents <1% of the world pharmaceutical market, with the pediatric market being a fraction of this. Thus, Australia's ability to influence the global market has been seen as limited, and it had been hoped that Australian children would benefit from international efforts in the area of medicines for children. This, however, has not eventuated, with there being very little change in the availability of medications for children in Australia over the past 2 decades. A number of reviews have been undertaken looking at the issue of medication for children in Australia and what could be done. Recently, two significant events have occurred: the first is the establishment of a pediatric medicines advisory group by the Australian government and the second is the decision to fund the development of a national pediatric dosing reference. These events represent significant progress, but are only the first of many steps required to improve medications for children in the 'land of Oz' (Australia). PMID- 19127950 TI - Pediatric clinical pharmacology studies in Chagas disease: focus on Argentina. AB - Chagas disease is a neglected parasitic disease endemic in the Americas. It mainly affects impoverished populations and the acute phase of the infection mostly affects children. Many cases have also been detected in nonendemic countries as a result of recent migratory trends. The chronic phase is relatively asymptomatic, but 30% of patients with chronic infection eventually develop cardiac and digestive complications that commonly lead to death or disability. Only two drugs are available for the treatment of Chagas disease, benznidazole and nifurtimox. These drugs have been shown to be effective in the treatment of both acute and early chronic phases in children, but the pharmacokinetics of these drugs have never been studied in this population. We have set out to conduct a pharmacokinetics study of benznidazole in a pediatric population with Chagas disease. The results of this study are expected to allow better estimation of the optimal doses and schedule of pharmacotherapy for Chagas disease in children. PMID- 19127952 TI - Delivering better medicines to children: need for better integration between the science, the policy, and the practice. AB - We are experiencing an exciting and unprecedented period in the history of children's medicines globally. Milestone developments unfolding in recent years include the formation of the International Alliance for Better Medicines for Children in 2006 and landmark initiatives by the EU, the World Health Assembly (WHA), and the WHO in 2007. However, the challenges of optimizing the development, wider availability, and routine use of effective, safe, and affordable medicines addressing important child health needs are considerable. Each aspect of this continuum has so far received differential attention. Major initiatives in the US and EU have focused on stimulating research into children's medicines, largely driven by drug regulatory reforms, but with important gaps remaining. Many countries are lacking similar reforms, so the benefits of these initiatives are currently not well reflected in the rest of the world. A systematic approach to knowledge translation to improve use of best evidence and deliver quality use of medicines (QUM) to children routinely has also been largely a 'missing link' so far. The WHO's Essential Medicines List for Children and related initiatives are addressing improving children's access to needed medicines. Priority research gaps, especially in the developing world, are also being pursued by the WHO. However, in many countries, including developed nations such as Australia, the policy response to the WHA resolution on 'Better Medicines for Children' has been inconsistent or fragmented. A better integrated overall approach, linking global medicines research efforts to child health needs and actual medicines use is needed. International networking to support the conduct, synthesis, and rapid dissemination of pediatric medicines research will help close knowledge gaps at a global level. Harmonization of pediatric drug regulation will support this goal and facilitate improving access to needed medicines. Increasing research into and dissemination of effective strategies to promote QUM is an essential component to maximize return (in health benefits) on increased investment in medicines research. A greater focus on QUM should also help create demand for better evidence from clinicians. Delivering on the promise of better medicines for children, wherever in the world they may be, depends on achieving successful integration between the science, the policy, and the practice of pediatric medicines. PMID- 19127953 TI - The National Institutes of Health and the Best Pharmaceuticals for Children Act. AB - The majority of drugs used to treat children are not labeled for use in children. The Best Pharmaceuticals for Children Act of 2002, re-authorized as the US FDA Amendments Act of 2007, directs the National Institutes of Health (NIH) to sponsor pediatric clinical trials of drugs lacking patent protection, if the FDA request for studies has been declined. The NIH is currently sponsoring 17 clinical studies. Challenges encountered include a paucity of investigators who are trained in pediatric clinical pharmacology; inadequate knowledge of the mechanisms of drug action in a growing child; and lack of pediatric formulations. PMID- 19127954 TI - Optimal drug therapy for children: Canadian initiatives. AB - Canada has a long tradition of undertaking research and training in pediatric clinical pharmacology, and has one of the longest continuous enterprises in this field in the world. Training in pediatric clinical pharmacology in Canada is nationally accredited and rigorous. Canada has the largest number of pediatric clinical pharmacologists per capita of any country on earth, but to date there have been no federally supported initiatives for child-focused drug research. The recent development of a unique national network focused on drug safety - the Genotypic Approaches to Therapy in Children - has provided a framework that it hopes will facilitate networking as well as the development of coordinated national and hopefully international initiatives in pediatric therapeutics. PMID- 19127955 TI - Medication errors in children. AB - Medication errors commonly involve children, with dosing errors being the most common. Medication errors are more frequent among the most sick patients who have urgent and complex medical conditions. Physicians who are less experienced, tired, depressed, and burnt out make more errors. The systems approach views every medical error as a system failure. The focus is on how to change the system in order to prevent errors. Adopting the systems approach will enhance patients' safety. Strategies that have been found to be effective in reducing medication errors include the use of computerized physician order entry systems, pre-printed order forms, color-coded systems, and involving pharmacists in clinical care. PMID- 19127956 TI - Pediatric formulations: international issues and potential solutions. AB - Appropriate dosage forms of medication are often not available for use in newborns, infants, and young children. This is a worldwide phenomenon, but especially in developing countries. The WHO's 'Make medicines child size' campaign emphasizes this shortcoming. Professional organizations, industry, and government from the international arena have the resources to address this and need to work together to create solutions. PMID- 19127957 TI - Challenges in pediatric drug development: a pharmaceutical industry perspective. AB - A paradigm change is taking place from protecting children against clinical research to protecting them through research. It is based on a better scientific understanding of the child's physiology, on the increasing potential of biomedical interventions, and on an evolving conviction of children's right to benefit from scientific and pharmaceutical progress. The WHO campaign 'Make medicines child size' is contributing to expand this to a global vision of the health of all children. Research-based pharmaceutical industry develops innovative new medicines for serious and life-threatening diseases. It has built up competency in pediatric drug development and has welcomed US and EU pediatric legislation as well as the WHO campaign. More diseases without effective therapy in the past will become treatable conditions. Eventually these advances will also be reflected in the medical care in developing countries. Research-based pharmaceutical industry can support specific pediatric aspects of drug development in neglected diseases by sharing its learning. The way forward will be a constructive dialog among the key stakeholders to ensure continuing improvement in worldwide child healthcare. PMID- 19127958 TI - Training clinicians in pediatric pharmacology-toxicology: the Toronto model. AB - Established in 1979, the Toronto Program in Pediatric Pharmacology has trained pediatricians and pharmacists from over 30 countries and five continents. Within the training guidelines of the Royal College of Physicians and Surgeons of Canada, the philosophy of the program is tailoring the goals to meet the specific needs and career goals of each trainee. The program will continue to prepare pediatricians for the rapidly changing challenges of rational drug therapy for children. PMID- 19127959 TI - Training pediatric clinical pharmacology and therapeutics specialists of the future: the needs, the reality, and opportunities for international networking. AB - In recent years there has been a rapid and marked increase in global recognition of the need for better medicines for children, with various initiatives being implemented at global and regional levels. These exciting developments are matched by recognition of the need to build greater capacity in the field of pediatric clinical pharmacology and therapeutics to help deliver on the promise of better medicines for children. A range of pediatric medicines researchers, educators, clinical therapeutics practitioners, and experts in drug evaluation, regulation, and broader medicines policy are needed on a larger scale, in both developed and developing world settings. The current and likely future training needs to meet these diverse challenges, the current realities of trying to meet such needs, and the opportunities for international networking to help meet future training needs are discussed from a global perspective. PMID- 19127960 TI - Clinical investigation in pediatrics: challenges and opportunities in Toronto, Canada. AB - We discuss the challenges and opportunities we face at The Hospital for Sick Children in Toronto, ON, Canada in conducting clinical investigation in children. Three areas are discussed: (i) the program structure of the Research Institute; (ii) the Toronto Centre for Phenogenomics as a common platform for preclinical studies across the university campus; and (iii) evolving clinical research infrastructure. Because preclinical and clinical investigation are becoming increasingly resource-intensive, we are witnessing a trend toward functional amalgamation of research teams, centralization of major experimental facilities, and the expansion of research infrastructure at local and national levels. Although these trends are common to clinical research in all age groups, pediatric investigation is further challenged by its relative paucity of research related human resources, and patient numbers at local and even national levels. To promote drug research in children, these organizational changes in the research framework need to be seen on a much broader international level. PMID- 19127961 TI - Physiology and experimental medicine: translational research at the Toronto Hospital For Sick Children. AB - Physiology and Experimental Medicine at The Hospital For Sick Children (Toronto, ON, Canada) is a multidisciplinary program that brings together significant interdisciplinary and interprofessional activity from different study areas such as: lung and cardiovascular physiology; imaging; metabolism and nutrition; pharmacology; and experimental medicine. The program is comprised of researchers, clinicians, and educators who focus on the study of complex, integrated bodily systems. Special emphasis is placed on investigations which bridge in vitro and in vivo approaches, basic science with clinical applications, experimental models with human disease, and human subject research. PMID- 19127962 TI - Training clinicians in Maternal-Fetal Pharmacology: closing a conceptual gap. AB - Developmental pharmacology is a science that does not start at birth, but rather preconceptionally. However, training for obstetricians, perinatologists, and neonatologists in this field is almost nonexistent. The novel Summer Institute in Maternal-Fetal Pharmacology is presented, with preliminary indications that it fulfils its mandate. PMID- 19127963 TI - Rotavirus vaccine RIX4414 (Rotarix): a review of its use in the prevention of rotavirus gastroenteritis. AB - Rotavirus vaccine RIX4414 is an oral vaccine composed of a monovalent, live, attenuated, human rotavirus strain of G1P[8] type. RIX4414 vaccination in infants aged 6-17 weeks at enrolment provided protection against rotavirus gastroenteritis (RVGE) of any severity and high-level protection against severe RVGE requiring hospitalization in large, randomized clinical trials conducted in a wide range of geographic regions. Protective efficacy was evident over the period (2 months) between the first and second doses of vaccine, and the protection afforded by the full two-dose course was sustained for at least 2 years, the limit to which efficacy was assessed. RIX4414 displayed protective efficacy against the common rotavirus G, P[8] types (G1P[8], G3P[8], G4P[8], and G9P[8]) and the fully heterotypic G2P[4] type. RIX4414 did not interfere with other common childhood injectable immunizations when administered concomitantly, suggesting that it should be possible to integrate the vaccine into most routine childhood vaccination schedules, including those still using oral poliovirus vaccine. RIX4414 was generally well tolerated and there was no evidence of an increased risk of intussusception. Although dependent on many factors, including prevalent infecting strains, efficacy rates, and vaccine costs, pharmacoeconomic analyses suggest that mass immunization with RIX4414 would be cost effective in many countries, especially when assessed from the societal perspective. Therefore, rotavirus vaccine RIX4414 offers a highly effective control strategy for reducing the burden of RVGE in infants. PMID- 19127964 TI - Kinetics of systemic cytokine and brain chemokine gene expression in murine toxoplasma infection. AB - Toxoplasma gondii often migrates to the central nervous system in immunocompromised patients, where it induces a severe inflammation referred to as Toxoplasma encephalitis. The mechanisms involved in control of parasite multiplication and prevention of Toxoplasma encephalitis remain unclear. The objective of the present study was to characterize the inflammatory response in the brains of mice during acute T. gondii infection, with emphasis on the expression of chemokine receptors. Susceptible C57BL/6 mice were orally infected with 10 cysts of the low-virulent ME49 strain of T. gondii. Levels of cytokines (TNF-alpha, IFN-gamma, IL-10, IL-6, and IL-12p70) and chemokines (CCL/2MCP-1) were measured in plasma at 5, 10, 15, 20, and 30 days after infection. In addition, the mRNA expression of chemokines (CCL5/RANTES, CCL2/MCP-1, CCL4/MIP 1beta) and chemokine receptors (CCR1, CCR2, CCR5, CCR7, CCR8, CXCR4, and CXR5) were measured in brain tissues at the same time points. Plasma levels of IFN gamma and CCL2/MCP-1 were highly expressed at day 5, whereas TNF-alpha had a moderate increase at day 5, peaked at day 10, and returned to normal levels by day 30. Plasma levels of IL-10, IL-6, and IL-12p70 were not detected throughout the study. Analyses of mRNA expression of chemokines and chemokine receptors in the brain showed that CCL5/ RANTES, CCR7, CXCR4, and CXCR5 were upregulated, peaking after 10 days of T. gondii infection. IgM-specific antibody levels increased at day 5 and peaked at days 10 and 30, whereas IgG levels increased at day 10 and continued to increase thereafter, reaching maximum levels at day 30 postinfection (PI). Our results suggest that T. gondii infection is controlled at local and systemic levels, and that proinflammatory proteins and their receptors may be acting coordinately to induce stage conversion and prevent parasite multiplication and development of Toxoplasma encephalitis. The early production of IFN-gamma and the delayed expression of CXCR4 and CXCR5 indicate that T. gondii induces an early robust cellular immune response, followed by a strong and sustained antibody-mediated immunity. PMID- 19127965 TI - Prevalence of Toxoplasma gondii antibodies in stray dogs in Taipei, Taiwan. AB - From October 2005 through September 2006, blood samples collected from 1,412 (768 male, 644 female) 1-yr-old and older stray dogs in Taipei City, Taiwan, were tested for antibodies to Toxoplasma gondii using the latex agglutination test (LAT). Antibodies (LAT titers >1:32) to T. gondii were found in 284 (20.1%) of the animals. Seroprevalences were not affected by the sex or density of dogs. The prevalence was highest in dogs from the eastern part of the city. PMID- 19127966 TI - Two new species of Camallanus (Nematoda: Camallanidae) from freshwater turtles in Queensland, Australia. AB - We describe 2 new species of Camallanus (Nematoda: Camallanidae) from freshwater turtles collected in Queensland, Australia: Camallanus nithoggi n. sp. from Elseya latisternum (Gray) and Camallanus waelhreow n. sp. from Emydura krefftii (Gray), Emydura macquarrii (Gray), and Em. macquarrii dharra Cann. The only Camallanus sp. previously reported from turtles is C. chelonius Baker, 1983 (all other species in the family have been transferred to Serpinema). The 2 new species described here differ from C. chelonius in the number of male preanal papillae (7 vs. 6 in C. chelonius), the number of male postanal papillae (5 vs. 4 in C. chelonius), and the number of buccal capsule ridges. Additionally, we removed the tissues overlying the buccal capsule and used scanning electron micrographs (SEM) to show that the peribuccal shields extend laterally from the buccal capsule, the basal ring is separated from the buccal capsule by a narrow isthmus, and there is a buttress along the lateral margin of the buccal capsule that has not previously been observed in species of Camallanus. PMID- 19127967 TI - In vitro hybridization of Haemoproteus spp.: an experimental approach for direct investigation of reproductive isolation of parasites. AB - In spite of their potential as model organisms and their medical importance, parasite speciation processes have been insufficiently discussed in the general literature. Scarcity of experimental data regarding reproductive isolation of parasites is a serious obstacle. Toward this end we developed a method for the investigation of hybridization of hemosporidian parasites in vitro. Five species of Haemoproteus (Haemosporida, Haemoproteidae) were isolated from naturally infected passerine birds. They were identified to species based on morphology of their gametocytes and PCR amplification of a segment of the parasite's mitochondrial cytochrome b gene. Hybridization of Haemoproteus spp. was initiated in vitro by mixing blood containing mature gametocytes of different species with a 3.7% solution of sodium citrate and exposure of the mixture to air. The following hybridization experiments were performed: (1) Haemoproteus minutus x Haemoproteus pallidus, (2) Haemoproteus balmorali x Haemoproteus tartakovskyi, and (3) Haemoproteus fringillae x H. tartakovskyi. The development of ookinetes of both species was blocked in the first experiment. Ookinetes of all species developed in other experiments, but presumed hybrids were distinguished only in the third experiment. Illustrations of ookinetes of all species are given. The present communication indicates that controlled experiments for direct hybridization of hemosporidians can be carried out in vitro. Such experimental research can be used to reconcile molecular and morphological data and to define biological species for this group of parasites. PMID- 19127968 TI - Wolbachia in Dirofilaria repens, an agent causing human subcutaneous dirofilariasis. AB - Human subcutaneous dirofilariasis is an increasingly reported zoonosis caused by several filarial species, in particular by Dirofilaria (Nochtiella) repens. Like many filarial worms, D. repens harbors the bacterial endosymbiont Wolbachia that has been implicated in the inflammatory features of filarial infection. Immunohistochemical staining against the Wolbachia surface protein (WSP) was carried out on 14 skin nodules and showed numerous bacteria within the intact worms and occasional positive staining within the surrounding inflammatory infiltrate. Serum samples from 11 of these patients resulted positive for total immunoglobulin G titers against WSP as examined in enzyme-linked immunosorbent assay. This is the first description of Wolbachia distribution in D. repens and the first report of specific immune response to Wolbachia in patients with subcutaneous dirofilariasis. PMID- 19127969 TI - Molecular and morphological analysis of Myxobolus spp. of salmonid fishes with the description of a new Myxobolus species. AB - While investigating the parasite fauna of wild coho salmon. Oncorhynchus kisutch (Walbaum, 1792), histological examination provided evidence of a new species of Myxobolus (Myxozoa: Myxosporea) infecting nerves of skeletal muscle. Spores were morphologically similar to those of the intramuscular Myxobolus insidiosus Wyatt and Pratt, 1963, both having pyriform spores with clavate polar capsules. However, the former developed exclusively in the nerves of skeletal muscle rather than in myocytes. We examined both species of Myxobolus derived from coho salmon; Chinook salmon, Oncorhynchus tshawytscha (Walbaum, 1792); cutthroat trout, Oncorhynchus clarkii (Richardson, 1836); and rainbow trout Oncorhynchus mykiss (Walbaum, 1792) from freshwater in Oregon. Spore morphology, small subunit ribosomal RNA gene (rDNA) sequences, and site of infection were compared. Myxobolus arcticus Pugachev and Khokhlov, 1979 has pyriform spores, infects the central nervous system of many salmonids, and is found in the Pacific Northwest. It was therefore included in the analyses to rule out conspecificity with the new species. Together, these data show that the Myxobolus sp. from peripheral nerves in the skeletal musculature of coho salmon, rainbow trout, and cutthroat trout is a new species, described herein as Myxobolus fryeri n. sp. PMID- 19127970 TI - Host susceptibility to schistosomes: effect of host sera on cell proliferation of Schistosoma mansoni schistosomula. AB - To determine effects of the sera on cell proliferation, schistosomula of Schistosoma mansoni (20-days-old) were incubated in medium containing fetal calf serum plus hamster (highly susceptible host) portal or peripheral venous serum, or rat (poorly susceptible host) portal or peripheral venous serum in the presence of bromodeoxyuridine (BrdU). Compared with schistosomula cultured in presence of control medium containing fetal calf serum alone, BrdU labeling indices (BLIs) were increased by 39% in the presence of portal, but not in peripheral, serum of hamsters. In contrast, no significant differences were observed in the BLIs in rat portal, or peripheral, sera or in control media. In vivo BrdU labeling results revealed that there was no detectable cell proliferation in S. mansoni schistosomula (6 days old) in the lungs. However, cell proliferation was detected in schistosomula beginning at 17 days. The results indicated that portal venous serum from a highly susceptible host, but not from a poorly susceptible host, stimulated schistosome cell proliferation in vitro. The timing of the increase in cell proliferation in terms of development corresponded to liver portal-mesenteric localization of schistosomula. Together, the data support the conclusion that in susceptible hosts, portal serum may play a role in schistosome cell proliferation, possibly resulting in termination of schistosome migration. This may explain the colocalization of adults, and the known organ selectivity of disease. PMID- 19127971 TI - A new species of mermithid (Nematoda) in chironomids eclosing from a northern Minnesota bog. AB - Hydromermis haggardi n. sp. (Nematoda: Mermithidae) is described from the hemocoel of chironomid imagos eclosing from Lake Alice Bog in northern Minnesota in June and July 2007. The species is distinguished from the other 27 described members of the genus by the subventral mouth, tulip shape to anterior end of esophagus, colorless vesicular trophosome, single nonbifurcated spicule, short body lengths of both males and females, structure of the S-shaped vagina, decrease in body width at vulva, brevity of postparasitic stage, and the greater development of a posterior ventral protractor muscle in the males. An anomalous adult male bearing large oval structures in the posterior testis is reported. Evidence is presented for the developmental interaction of the new species with Lanceimermis palustris Johnson and Kleve, 2004. PMID- 19127974 TI - Time to move from presumptive malaria treatment to laboratory-confirmed diagnosis and treatment in African children with fever. AB - BACKGROUND TO THE DEBATE: Current guidelines recommend that all fever episodes in African children be treated presumptively with antimalarial drugs. But declining malarial transmission in parts of sub-Saharan Africa, declining proportions of fevers due to malaria, and the availability of rapid diagnostic tests mean it may be time for this policy to change. This debate examines whether enough evidence exists to support abandoning presumptive treatment and whether African health systems have the capacity to support a shift toward laboratory-confirmed rather than presumptive diagnosis and treatment of malaria in children under five. PMID- 19127975 TI - Effect of removing direct payment for health care on utilisation and health outcomes in Ghanaian children: a randomised controlled trial. AB - BACKGROUND: Delays in accessing care for malaria and other diseases can lead to disease progression, and user fees are a known barrier to accessing health care. Governments are introducing free health care to improve health outcomes. Free health care affects treatment seeking, and it is therefore assumed to lead to improved health outcomes, but there is no direct trial evidence of the impact of removing out-of-pocket payments on health outcomes in developing countries. This trial was designed to test the impact of free health care on health outcomes directly. METHODS AND FINDINGS: 2,194 households containing 2,592 Ghanaian children under 5 y old were randomised into a prepayment scheme allowing free primary care including drugs, or to a control group whose families paid user fees for health care (normal practice); 165 children whose families had previously paid to enrol in the prepayment scheme formed an observational arm. The primary outcome was moderate anaemia (haemoglobin [Hb] < 8 g/dl); major secondary outcomes were health care utilisation, severe anaemia, and mortality. At baseline the randomised groups were similar. Introducing free primary health care altered the health care seeking behaviour of households; those randomised to the intervention arm used formal health care more and nonformal care less than the control group. Introducing free primary health care did not lead to any measurable difference in any health outcome. The primary outcome of moderate anaemia was detected in 37 (3.1%) children in the control and 36 children (3.2%) in the intervention arm (adjusted odds ratio 1.05, 95% confidence interval 0.66 1.67). There were four deaths in the control and five in the intervention group. Mean Hb concentration, severe anaemia, parasite prevalence, and anthropometric measurements were similar in each group. Families who previously self-enrolled in the prepayment scheme were significantly less poor, had better health measures, and used services more frequently than those in the randomised group. CONCLUSIONS: In the study setting, removing out-of-pocket payments for health care had an impact on health care-seeking behaviour but not on the health outcomes measured. PMID- 19127976 TI - Abolishing user fees in Africa. PMID- 19127977 TI - Abandoning presumptive antimalarial treatment for febrile children aged less than five years--a case of running before we can walk? AB - BACKGROUND TO THE DEBATE: Current guidelines recommend that all fever episodes in African children be treated presumptively with antimalarial drugs. But declining malarial transmission in parts of sub-Saharan Africa, declining proportions of fevers due to malaria, and the availability of rapid diagnostic tests mean it may be time for this policy to change. This debate examines whether enough evidence exists to support abandoning presumptive treatment and whether African health systems have the capacity to support a shift toward laboratory-confirmed rather than presumptive diagnosis and treatment of malaria in children under five. PMID- 19127978 TI - An endoribonuclease functionally linked to perinuclear mRNP quality control associates with the nuclear pore complexes. AB - Nuclear mRNA export is a crucial step in eukaryotic gene expression, which is in yeast coupled to cotranscriptional messenger ribonucleoprotein particle (mRNP) assembly and surveillance. Several surveillance systems that monitor nuclear mRNP biogenesis and export have been described, but the mechanism by which the improper mRNPs are recognized and eliminated remains poorly understood. Here we report that the conserved PIN domain protein Swt1 is an RNA endonuclease that participates in quality control of nuclear mRNPs and can associate with the nuclear pore complex (NPC). Swt1 showed endoribonuclease activity in vitro that was inhibited by a point mutation in the predicted catalytic site. Swt1 lacked clear sequence specificity but showed a strong preference for single-stranded regions. Genetic interactions were found between Swt1 and the THO/TREX and TREX-2 complexes, and with components of the perinuclear mRNP surveillance system, Mlp1, Nup60, and Esc1. Inhibition of the nuclease activity of Swt1 increased the levels and cytoplasmic leakage of unspliced aberrant pre-mRNA, and induced robust nuclear poly(A)(+) RNA accumulation in mlp1Delta and esc1Delta strains. Overexpression of Swt1 also caused strong nuclear poly(A)(+) RNA accumulation. Swt1 is normally distributed throughout the nucleus and cytoplasm but becomes concentrated at nuclear pore complexes (NPCs) in the nup133Delta mutant, which causes NPC clustering and defects in mRNP export. The data suggest that Swt1 endoribonuclease might be transiently recruited to NPCs to initiate the degradation of defective pre-mRNPs or mRNPs trapped at nuclear periphery in order to avoid their cytoplasmic export and translation. PMID- 19127979 TI - Collective cell migration drives morphogenesis of the kidney nephron. AB - Tissue organization in epithelial organs is achieved during development by the combined processes of cell differentiation and morphogenetic cell movements. In the kidney, the nephron is the functional organ unit. Each nephron is an epithelial tubule that is subdivided into discrete segments with specific transport functions. Little is known about how nephron segments are defined or how segments acquire their distinctive morphology and cell shape. Using live, in vivo cell imaging of the forming zebrafish pronephric nephron, we found that the migration of fully differentiated epithelial cells accounts for both the final position of nephron segment boundaries and the characteristic convolution of the proximal tubule. Pronephric cells maintain adherens junctions and polarized apical brush border membranes while they migrate collectively. Individual tubule cells exhibit basal membrane protrusions in the direction of movement and appear to establish transient, phosphorylated Focal Adhesion Kinase-positive adhesions to the basement membrane. Cell migration continued in the presence of camptothecin, indicating that cell division does not drive migration. Lengthening of the nephron was, however, accompanied by an increase in tubule cell number, specifically in the most distal, ret1-positive nephron segment. The initiation of cell migration coincided with the onset of fluid flow in the pronephros. Complete blockade of pronephric fluid flow prevented cell migration and proximal nephron convolution. Selective blockade of proximal, filtration-driven fluid flow shifted the position of tubule convolution distally and revealed a role for cilia-driven fluid flow in persistent migration of distal nephron cells. We conclude that nephron morphogenesis is driven by fluid flow-dependent, collective epithelial cell migration within the confines of the tubule basement membrane. Our results establish intimate links between nephron function, fluid flow, and morphogenesis. PMID- 19127980 TI - Regional risk assessment as a part of the long-term receiving water study. AB - A regional-scale watershed assessment using the relative risk model has now been performed on each of the watersheds that are part of the Long-Term Receiving Waters Study (LTRWS) to generate patterns of risk at each study site. The watersheds assessed included the lower McKenzie and mid-Willamette rivers in Oregon, USA; Codorus Creek in Pennsylvania, USA; and the Leaf River in Mississippi, USA. In each case, the goal was to put the LTRWS into a watershed and regional context, including multiple sources, stressors, habitats, and assessment endpoints. In each instance, the relative risk model provided measures of relative risk and testable hypotheses about patterns within the watersheds. There were similarities among the 3 risk studies. First, land use within the watershed was the most important driver of risk in each instance, even compared with point sources. Second, the list of endpoints was similar for each risk assessment. Water quality was held as important, along with populations critical to recreation or commercial use. At each site, data from the LTRWS and other sources could be used to support the risk patterns predicted from the risk assessment process. However, the sampling design of the LTRWS prevented the confirmation of specific cause-effect relationships. A fundamental conclusion is that risk assessment, using an adaptation of the relative risk model, should be performed as an integral part of any watershed assessment and management effort. These initial watershed risk assessments have led to similar assessments across the world. PMID- 19127981 TI - Patterns of macroinvertebrate assemblages in a long-term watershed-scale study to address the effects of pulp and paper mill discharges in four US receiving streams. AB - Changes in macroinvertebrate communities exposed to pulp and paper mill effluent (PPME) have been seen in mesocosm and short-term field studies. However, long term patterns of macroinvertebrates in PPME receiving streams have not been examined. We conducted a study of 4 PPME receiving streams (Codorus Creek, PA; the Leaf River, MS; and the McKenzie and Willamette rivers, OR) over 9 y to assess temporal patterns in macroinvertebrate community structure and metrics related to PPME discharge. Study streams represented different ecoregions, warm /cold-water systems, gradients of PPME concentration (<1%-33%), and mill process types. Bray-Curtis similarity and nonmetric multidimensional scaling showed significant community differences across sites in Codorus Creek, but differences were related to stream temperature patterns and not PPME. In the other study streams, seasonal community differences across years were greater than differences across sites. General linear models were used to examine spatial and temporal variation in macroinvertebrate metric response (% dominant taxa, density, richness, Hilsenhoff Biotic Index [HBI], Simpson's Index, and ash-free dry mass). Mean HBI scores indicated that the macroinvertebrate community reflected fair to very good water quality conditions, with water quality typically classified as "good" at most sites. Significant site differences in macroinvertebrate metric response were uncommon in the Leaf, McKenzie, and Willamette rivers but were seen in all metrics in Codorus Creek, where metric response was spatially variable. In the McKenzie River, there was an increase in mean HBI scores at sites downstream of the mill relative to 1 of the 2 upstream sites. However, significant differences were seen only between 1 upstream and downstream site, and HBI scores at all downstream sites consistently reflected "good" water quality. Significant annual differences in metric response were typical in all rivers. Water quality (pH, conductivity, total nitrogen) and habitat (velocity, depth, substrate composition) variables explained community structure patterns in the Leaf and McKenzie rivers, but macroinvertebrate environment relationships were weak in the other 2 streams. The results of this study indicate that macroinvertebrate community structure is temporally variable and reiterate the importance of long-term studies for accurate determination of the effects of point sources such as PPME on receiving systems. PMID- 19127982 TI - Fragment molecular orbital calculations on red fluorescent proteins (DsRed and mFruits). AB - We have performed a series of fragment molecular orbital (FMO) calculations for a family of red fluorescent proteins, DsRed and mFruits. The electronic transition energies were evaluated by the method of configuration interaction singles with perturbative doubles [CIS(D)] including higher-order corrections. The calculated values were in good agreement with the corresponding experimental peak values of spectra. Additionally, the chromophore environment was systematically analyzed in terms of the interaction energies between the pigment moiety and neighboring residues. It was theoretically revealed that the electrostatic interactions play a dominant role in the DsRed chromophore, whereas the color tunings in mFruits are controlled in a more delicate fashion. PMID- 19127983 TI - Molecular simulation of excess isotherm and excess enthalpy change in gas-phase adsorption. AB - We present a new approach to calculating excess isotherm and differential enthalpy of adsorption on surfaces or in confined spaces by the Monte Carlo molecular simulation method. The approach is very general and, most importantly, is unambiguous in its application to any configuration of solid structure (crystalline, graphite layer or disordered porous glass), to any type of fluid (simple or complex molecule), and to any operating conditions (subcritical or supercritical). The behavior of the adsorbed phase is studied using the partial molar energy of the simulation box. However, to characterize adsorption for comparison with experimental data, the isotherm is best described by the excess amount, and the enthalpy of adsorption is defined as the change in the total enthalpy of the simulation box with the change in the excess amount, keeping the total number (gas + adsorbed phases) constant. The excess quantities (capacity and energy) require a choice of a reference gaseous phase, which is defined as the adsorptive gas phase occupying the accessible volume and having a density equal to the bulk gas density. The accessible volume is defined as the mean volume space accessible to the center of mass of the adsorbate under consideration. With this choice, the excess isotherm passes through a maximum but always remains positive. This is in stark contrast to the literature where helium void volume is used (which is always greater than the accessible volume) and the resulting excess can be negative. Our definition of enthalpy change is equivalent to the difference between the partial molar enthalpy of the gas phase and the partial molar enthalpy of the adsorbed phase. There is no need to assume ideal gas or negligible molar volume of the adsorbed phase as is traditionally done in the literature. We illustrate this new approach with adsorption of argon, nitrogen, and carbon dioxide under subcritical and supercritical conditions. PMID- 19127986 TI - Roles of the ether oxygen in hydration of tetrahydrofuran studied by IR, NMR, and DFT calculation methods. AB - We studied the concentration dependence of nu(C-H)'s in IR and (1)J(C,H) in NMR for binary water-tetrahydrofuran (THF) mixtures and found different trends for the two types of CH(2) groups in the five-membered ring. The changes of the nu(C O) spectra showed that complexes of THF associated with water are formed, in which the number of water molecules increases with the water concentration. We suggested that hydration proceeds through the formation of 1:1, and 1:2 complexes of [THF:water] up to X(H(2)O) approximately 0.9, where X(H)((2))(O) is the mole fraction of the water in the mixtures. We carried out ab initio MO and DFT calculations to optimize the geometries of a THF dimer as a model of THF molecules in pure liquid, and 1:1 and 1:2 complexes of [THF:water] to simulate observed concentration dependence of nu(C-H)'s in IR and (1)J(C,H) in NMR. The changes of the calculated nu(C-H) spectra and (1)J(C,H) values for the optimized complexes are in agreement with those observed with varying X(H)((2))(O), supporting our proposal. From the vibrational and NBO analyses of the optimized complexes, the observed blue shift of nu(C-H)'s and the increase of (1)J(C,H) for the CH(2) groups neighboring to the ether oxygen were explained in terms of the changes in the stereoelectronic effect, resulting from HO-H...O< hydrogen bonding. The optimized 1:2-complex contains two weak C-H...OH(2) hydrogen bonds, and blue shift of nu(C-H)'s and increase of (1)J(C,H) were demonstrated from the same analyses of the complexes. This result of simulation also supports that the blue shift of nu(C-H)'s and increase of (1)J(C,H) observed for both the type of CH(2) groups at 0.6 X(H)((2))(O) < 0.9 are attributed to these interactions. On the basis of all these results, we propose that the formation of the 1:2-complex involving weak C-H...OH(2) hydrogen bonds is responsible dominantly for the hydrophobic hydration of THF. PMID- 19127987 TI - Bases for Understanding Polymerization under Pressure: The Practical Case of CO2. AB - We present a novel quantitative strategy for monitoring chemical bonding transformations in solids from the topology of their electronic structure. Developed in the context of the electron localization function formalism, it provides an unambiguous characterization of long-range interactions and bond formation. Charge flux between electron localization regions is found to hold the key for identifying the nature of the interaction between the chemically meaningful entities in the solid (valence shells, lone pairs, molecules, etc.). Because of the wide range of interesting properties that high pressure induces in molecular solids, we illustrate the potentialities of our strategy to unveil controversial questions involved in the bond reorganization along the polymerization of CO2. Our study confirms that the topology of the bonding network in the pseudopolymeric phases points toward the incipient formation of the new bonds in the higher pressure polymers. This transformation is identified as a synchronic weakening of the intramolecular (C==O) double bond and the birth of a new intermolecular C--O bond controlled by the oxygen lone pairs. Overall, the relationship that this type of analysis establishes between different polymorphs of the phase diagram could be further exploited for the prediction of the coordination of high pressure phases, opening new avenues for experimental synthesis and structure indexation. PMID- 19127989 TI - Rate constants for the gas-phase reactions of OH radicals with a series of C6-C14 alkenes at 299 +/- 2 K. AB - Rate constants for the gas-phase reactions of OH radicals with the C(6)-C(14) 2 methyl-1-alkenes and the C(6)-C(10) trans-2-alkenes have been measured at 299 +/- 2 K and atmospheric pressure of air using a relative rate technique. The rate constants obtained (in units of 10(-11) cm(3) molecule(-1) s(-1)) were as follows: 2-methyl-1-pentene, 5.67 +/- 0.21; 2-methyl-1-hexene, 6.50 +/- 0.11; 2 methyl-1-heptene, 6.71 +/- 0.21; 2-methyl-1-octene, 7.02 +/- 0.16; 2-methyl-1 nonene, 7.28 +/- 0.21; 2-methyl-1-decene, 7.85 +/- 0.26; 2-methyl-1-undecene, 7.85 +/- 0.21; 2-methyl-1-dodecene, 7.96 +/- 0.26; 2-methyl-1-tridecene, 8.06 +/- 0.37; trans-2-hexene, 6.08 +/- 0.26; trans-2-heptene, 6.76 +/- 0.32; trans-2 octene, 7.23 +/- 0.21; trans-2-nonene, 7.54 +/- 0.16; and trans-2-decene, 7.80 +/ 0.26, where the indicated errors are two least-squares standard deviations and do not include the uncertainty associated with the rate constant for the reference compound alpha-pinene. Our data show that the rate constants for the reactions of OH radicals with 2-methyl-1-alkenes and trans-2-alkenes increase with increasing carbon number, suggesting that this is in part due to H-atom abstraction from the C-H bonds of the alkyl substituent groups. Combined with previous literature data for the reactions of OH radicals with a series of 1 alkenes, we propose that the increase in rate constant with increasing carbon number is due to H-atom abstraction from the C-H bonds of the alkyl substituent groups and to enhancement of the rate constant for OH radical addition to the C=C bond, which increases with carbon number of a C(n)-alkyl substituent group up to a maximum at approximately C(8). PMID- 19127988 TI - Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins. AB - Fluorescent proteins with long emission wavelengths are particularly attractive for deep tissue two-photon microscopy. Surprisingly, little is known about their two-photon absorption (2PA) properties. We present absolute 2PA spectra of a number of orange and red fluorescent proteins, including DsRed2, mRFP, TagRFP, and several mFruit proteins, in a wide range of excitation wavelengths (640-1400 nm). To evaluate 2PA cross section (sigma(2)), we use a new method relying only on the optical properties of the intact mature chromophore. In the tuning range of a mode-locked Ti:sapphire laser, 700-1000 nm, TagRFP possesses the highest two photon cross section, sigma(2) = 315 GM, and brightness, sigma(2)phi = 130 GM, where phi is the fluorescence quantum yield. At longer wavelengths, 1000-1100 nm, tdTomato has the largest values, sigma(2) = 216 GM and sigma(2)phi = 120 GM, per protein chain. Compared to the benchmark EGFP, these proteins present 3-4 times improvement in two-photon brightness. PMID- 19127990 TI - The Groningen Protocol: is it necessary? Is it scientific? Is it ethical? AB - AIM: To explore whether the Groningen Protocol is clinically necessary, scientific, and ethically justified. RESULTS: The Groningen Protocol is clinically unnecessary because the withdrawal of life-sustaining treatment from seriously ill infants is already ethically accepted globally without the need for the Protocol and because spina bifida, to which the Protocol has been most often applied, can be detected by ultrasound before viability, affording pregnant women the opportunity of termination of pregnancy. The Groningen Protocol is unscientific because it does not meet the accepted standards of evidence-based reasoning concerning the four clinical criteria for its application. The Groningen Protocol is unethical because it does not meet the standards of argument-based ethics in defining its four clinical criteria. CONCLUSIONS: The Groningen Protocol is clinically unnecessary, unscientific, and unethical. Physicians should condemn its use. The Dutch Association of Paediatrics should revoke its approval and adoption of the Groningen Protocol. PMID- 19127991 TI - Sensing polycyclic aromatic hydrocarbons with dithiocarbamate-functionalized ag nanoparticles by surface-enhanced Raman scattering. AB - Trace detection of polycyclic aromatic hydrocarbons is reported in this work on dithiocarbamate calix[4]arene functionalized Ag nanoparticles by using surface enhanced Raman scattering (SERS). SERS spectra informed about the existence of the pollutant by measuring its characteristic fingerprint vibrational features. In addition, SERS revealed important structural information from both the host and the analyte which was crucial to understand and deduce the host-guest interaction mechanism. The effectiveness of this system was checked for a group of PAHs: pyrene, benzo[c]phenanthrene, triphenylene, and coronene. From the analyzed results, the affinity constants and the limit of detection were deduced for each pollutant. PMID- 19127993 TI - Solid-phase synthesis of 1,3,6-trisubstituted-1H-thiazolo[4,5-c][1,2]thiazin 4(3H)one-2,2-dioxide derivatives using traceless linker. AB - A new solid-phase route for preparation of 1,3,6-trisubstituted-1H-thiazolo[4,5 c][1,2]thiazin-4(3H)one-2,2-dioxide derivatives is described. Our synthetic route is begun with a thiazole resin and relies on the sulfonamide formation, Mitsunobu type N-alkylation, cyclization, and nucleophilic substitution methodology cleavage on a solid support. The strategy permits the incorporation of three points of diversity into the thiazolo[4,5-c][1,2]thiazine ring system in good overall yields. PMID- 19127992 TI - Single myosin lever arm orientation in a muscle fiber detected with photoactivatable GFP. AB - Myosin 2 is the molecular motor in muscle. It binds actin and executes a power stroke by rotating its lever arm through an angle of approximately 70 degrees to translate actin against resistive force. Myosin 2 has evolved to function optimally under crowded conditions where rates and equilibria of macromolecular reactions undergo major shifts relative to those measured in dilute solution. Hence, an important research objective is to detect in situ the lever arm orientation. Single-molecule measurements are preferred because they clarify ambiguities that are unavoidable with ensemble measurements; however, detecting single molecules in the condensed tissue medium where the myosin concentration exceeds 100 muM is challenging. A myosin light chain (MLC) tagged with photoactivatable green fluorescent protein (PAGFP) was constructed. The recombinant MLC physically and functionally replaced native MLC on the myosin lever arm in a permeabilized skeletal muscle fiber. Probe illumination volume was minimized using total internal reflection fluorescence microscopy, and PAGFP was sparsely photoactivated such that polarized fluorescence identified a single probe orientation. Several physiological states of the muscle fiber were characterized, revealing two distinct orientation populations in all states called straight and bent conformations. Conformation occupancy probability varies among fiber states with rigor and isometric contraction at extremes where straight and bent conformations predominate, respectively. Comparison to previous work on single rigor cross-bridges at the A-band periphery where the myosin concentration is low suggests molecular crowding in the A-band promotes occupancy of the straight myosin conformation [Burghardt, T. P., et al. (2007) Biophys. J. 93, 2226]. The latter may have a role in contraction because it provides additional free energy favoring completion of the cross-bridge power stroke. PMID- 19127994 TI - Peculiar high-pressure behavior of BiMnO3. AB - High-pressure structural properties of perovskite-type BiMnO(3) have been investigated by synchrotron X-ray powder diffraction at room temperature. A new monoclinic phase having P2(1)/c symmetry was found between about 1.5 and 5.5 GPa. Above 8 GPa, the orthorhombic GdFeO(3)-type phase (space group Pnma) is stable. The crystal structure of BiMnO(3) at 8.6 GPa and room temperature was investigated (a = 5.5132(3) A, b = 7.5752(3) A, c = 5.4535(3) A). The orthorhombic phase of BiMnO(3) has an orbital order similar to LaMnO(3) but with a different arrangement of orbitals in the ac plane. High-pressure room temperature behavior of BiMnO(3) differs from high-temperature behavior at ambient pressure in comparison with BiCrO(3) and BiScO(3). These findings may open new directions in investigation of BiMnO(3). PMID- 19127995 TI - An analysis of substituent effects in ethane derivatives: the quantum theory of atoms in molecules approach. AB - MP2/6-311++G(d,p) calculations were performed on ethane and its simple derivatives. Different substituents such as: Li, Na, BeH, BH(2), OH, NH(2), F and NO(2) were considered. The mono- and disubstituted derivatives of ethane were analyzed. The quantum theory of "atoms in molecules" was applied to localize bond critical points and to analyze relationships between their locations and characteristics of the species. The numerous correlations were found showing the crucial influence of substituent effects. It was also revealed that such parameters as the carbon-carbon distance, or the electron density at C-C bond critical point do not show the systematic changes caused by substituent effects. This means that some of QTAIM parameters, especially the positions of BCPs may be useful to detect numerous sensitive effects if the latter ones do not affect the traditionally analyzed geometrical parameters of a molecule, such as bond lengths. PMID- 19127996 TI - Ultrafast FRET in a room temperature ionic liquid microemulsion: a femtosecond excitation wavelength dependence study. AB - Fluorescence resonance energy transfer (FRET) from coumarin 480 (C480) to rhodamine 6G (R6G) is studied in a room temperature ionic liquid (RTIL) microemulsion by picosecond and femtosecond emission spectroscopy. The microemulsion is comprised of the RTIL 1-pentyl-3-methylimidazolium tetraflouroborate, [pmim][BF4], in TX-100/ benzene. We have studied the microemulsion with and without water. The time constants of FRET were obtained from the risetime of the acceptor (R6G) emission. In the RTIL microemulsion, FRET occurs on multiple time scales: 1, 250, and 3900 ps. In water containing RTIL microemulsion, the rise components are 1.5, 250, and 3900 ps. The 1 and 1.5 ps components are assigned to FRET at a close contact of donor and acceptor (RDA approximately 12 A). This occurs within the highly polar (RTIL/water) pool of the microemulsion. With increase in the excitation wavelength (lambdaex) from 375 to 435 nm, the relative contribution of the ultrafast component of FRET (1 ps) increases from 4% to 100% in the RTIL microemulsion and 12% to 100% in the water containing RTIL microemulsion. It is suggested that at lambdaex = 435 nm, mainly the highly polar RTIL pool is probed where FRET is very fast due to the close proximity of the donor and the acceptor. The very long 3900 ps (RDA approximately 45 A) component may arise from FRET from a donor in the outer periphery of the microemulsion to an acceptor in the polar RTIL pool. The 250 ps component (RDA approximately 29 A) is assigned to FRET from a donor inside the surfactant chains. PMID- 19127997 TI - Water-dichloromethane interface controlled synthesis of hierarchical rutile TiO2 superstructures and their photocatalytic properties. AB - A water-dichloromethane interface is used for synthesis and assembly of rutile TiO(2) nanorods. By hydrothermal treatment of a dichloromethane solution of TiCl(4) at the interface of water-dichloromethane, turning to no surfactant or template, hierarchical rutile TiO(2) superstructures are developed. By tuning the molar ratio of reactants r(w) (H(2)O/TiCl(4)), the size and shape of the samples significantly change. At a low value of r(w), highly extended, robust, porous, and thick titania film with ordered rutile nanorod bundles are deposited at the interface. At a high value of r(w), powders consisting of hierarchical rutile nanorod spheres together with disordered nanorods are obtained. A rational formation mechanism is proposed on the basis of a range of experiments. The main factors influencing the morphologies of the samples may be attributed to the acidity of the reaction system and the adsorption ability of the precursor nanoparticles to the water-dichloromethane interface. The as-obtained rutile TiO(2) hierarchical superstructures show higher photocatalytic property to decompose methylene blue (MB) dye compared with that of commercial P25, which can be ascribed to the contribution of high surface area and high crystallinity. Other applications, such as solar energy conversion, environmental remediation, and advanced optical/electric nanodevices may also benefit from the unique properties of the hierarchically rutile TiO(2) superstructures. PMID- 19127998 TI - Model Ziegler-type hydrogenation catalyst precursors, [(1,5-COD)M(mu-O2C8H15)]2 (M = Ir and Rh): synthesis, characterization, and demonstration of catalytic activity en route to identifying the true industrial hydrogenation catalysts. AB - The compounds [(1,5-COD)M(mu-O2C8H15)]2 (COD = cyclooctadiene, M = Ir (1) or Rh (2), O2C8H15 = 2-ethylhexanoate) were synthesized by addition of Bu3NH(2 ethylhexanoate) or Na(2-ethylhexanoate) to acetone suspensions of [(1,5-COD)Ir(mu Cl)]2 or [(1,5-COD)Rh(mu-Cl)]2, respectively. The synthesis of such well-defined second and third row model precursors is key to determining the true nature of commercial Ziegler-type hydrogenation catalysts (i.e., catalysts made from the combination of a non-zerovalent, group 8-10 transition metal precatalyst and a trialkylaluminum cocatalyst), an unsolved, approximately 40 year old problem. The characterizations of 1 and 2 were accomplished by elemental analysis, melting point, FAB-MS, FT-IR, UV-vis, NMR spectroscopy, and single crystal X-ray diffraction. The complexes, C32H54Ir2O4 and C32H54O4Rh2, are isostructural: monoclinic, P2(1)/n, Z = 4. The lattice constants for 1 are a = 15.7748(5) A, b = 9.8962(3) A, c = 20.8847(7) A, beta = 108.408(2) degrees. The lattice constants for 2 are a = 15.7608(4) A, b = 9.9032(3) A, c = 20.8259(5) A, beta = 108.527(1) degrees. Complexes 1 and 2 are dimeric, bridged by the 2-ethylhexanoates, and with one 1,5-COD ligand bound to each metal. The formally 16 electron metal atoms are in square ligand planes with dihedral angles between the planes of 56.5 degrees for 1 and 58.1 degrees for 2. The M-M distances of 3.2776(2) and 3.3390(4) A for 1 and 2, respectively, fall in the range of similar structures thought to have some M-M interaction despite the lack of a formal M-M bond. Demonstration that active Ziegler-type hydrogenation catalysts are made when 1 or 2 combine with AlEt3 is provided, results that open the door to the use of 1 and 2 as well-defined third and second row congeners, respectively, of Ziegler-type hydrogenation catalysts. These compounds have proven important in addressing the previously unsolved problem of the true nature of the catalyst in industrial Ziegler-type hydrogenation catalyst systems; their high yield synthesis and unequivocal characterization reported herein are the necessary first steps of that work. PMID- 19127999 TI - Vanadium-induced nucleophilic IPSO substitutions in a coordinated tetrachlorosemiquinone ring: formation of the chloranilate anion as a bridging ligand. AB - In basic media, the coordinated semiquinone radical in the spin-coupled [(bipy)ClV(IV)O(TCSQ)] 1 (HTCSQ = tetrachlorosemiquinone) undergoes nucleophilic ipso substitution (OH- for Cl-) to generate the chloranilate anion (CA(2-)) that bridges the vanadium(IV) centers, forming a binuclear compound [(bipy)ClV(IV)O(CA)OV(IV)Cl(bipy)] 2. PMID- 19128000 TI - Site-specific immobilization and micrometer and nanometer scale photopatterning of yellow fluorescent protein on glass surfaces. AB - A simple method is described for the site-specific attachment of yellow fluorescent protein (YFP) to glass surfaces on length scales ranging from tens of micrometers to ca. 200 nm. 3-Mercaptopropyl(triethoxy silane) is adsorbed onto a glass substrate and subsequently derivatized using a maleimide-functionalized oligomer of ethylene glycol. The resulting protein-resistant surface is patterned by exposure to UV light, causing photochemical degradation of the oligo(ethylene glycol) units to yield aldehyde groups in exposed regions. These are covalently bound to N-(5-amino-1-carboxypentyl)iminoacetic acid, yielding a nitrilotriacetic acid (NTA)-functionalized surface, which following complexation with Ni(2+), is coupled to His-tagged YFP. Using scanning near-field photolithography, in which a UV laser coupled to a scanning near-field optical microscope is utilized as the light source for photolithography, it is possible to fabricate lines of protein smaller than 200 nm, in which the biomolecules remain strongly optically active, facilitating the acquisition of diffraction-limited fluorescence images by confocal microscopy. PMID- 19128001 TI - 3-D lanthanide metal-organic frameworks: structure, photoluminescence, and magnetism. AB - A series of isostructural three-dimensional metal-organic frameworks [Pr(2)(N BDC)(3)(dmf)(4)](infinity) (1), {[Eu(2)(N-BDC)(3)(dmf)(4)] x 2DMF}(infinity) (2 x 2DMF), [Gd(2)(N-BDC)(3)(dmf)(4)](infinity) (3), {[Tb(2)(N-BDC)(3)(dmf)(4)] x 2DMF}(infinity) (4 x 2DMF), {[Dy(2)(N-BDC)(3)(dmf)(4)] x 2DMF}(infinity) (5 x 2DMF) (N-H(2)BDC = 2-amino-1,4-benzenedicarboxylic acid; DMF = N,N' dimethylformamide) with cubic 4(12) x 6(3) topology have been synthesized using solvothermal conditions. The networks were generated via formation of a dinuclear Ln(2) secondary building block, involving the dicarboxylate ligand as a bridge. The luminescent properties of the Tb(III) and Eu(III) complexes were studied and showed characteristic emissions at room temperature. Antiferromagnetic interactions between Ln(III) ions were observed from magnetic susceptibility data. PMID- 19128002 TI - Molecular dynamics simulations of methane hydrate decomposition. AB - Molecular dynamics simulations have been carried out to study decomposition of methane hydrate at different cage occupancies. The decomposition rate is found to depend sensitively on the hydration number. The rate of the destruction of the cages displays Arrhenius behavior, consistent with an activated mechanism. During the simulations, reversible formation of partial water cages around methane molecules in the liquid was observed at the interface at temperatures above the computed hydrate decomposition temperature. PMID- 19128003 TI - Diquaternary ammonium compounds in zeolite synthesis: cyclic and polycyclic N heterocycles connected by methylene chains. AB - An additional dimension has been added to our long-standing studies in high silica zeolite synthesis via a guest/host synergism. We have created and studied the impact of making symmetric diquaternary ammonium compounds, by varying the chain length between nitrogen charge centers, and the heterocycle size and geometry containing the nitrogen. This allows the introduction of a second spatial parameter in the use of the charged organo-cation guest in the zeolite synthesis. The series of 15 diquaternary ammonium compounds (5 heterocycles synthesized onto chain lengths of C4-C6) were tested in a total of 135 zeolite syntheses reactions. Nine screening reactions were employed for each guest molecule, and the conditions built upon past successes in finding novel high silica zeolites via introduction of boron, aluminum, or germanium as substituting tetrahedral framework atoms for silicon. Eighteen different zeolite structures emerged from the studies. The use of specific chain lengths for derivatives of the pyrrolidine ring system produced novel zeolite materials SSZ-74 and 75. PMID- 19128004 TI - High yield preparation of macroscopic graphene oxide membranes. AB - Graphene oxide membranes up to 2000 microm(2) in size can be synthesized with 90% yield in bulk quantities through a microwave-assisted chemical method. Membranes are readily visualized on an oxidized silicon substrate, which enables efficient fabrication of electronic devices and sensors. Field effect transistors made of the membrane show ambipolar behavior, and their conductivity is significantly higher than previously reported values. PMID- 19128005 TI - Comparison of quantum dot biodistribution with a blood-flow-limited physiologically based pharmacokinetic model. AB - A physiologically based pharmacokinetic model with partition coefficients estimated from quantum dot (QD) 705 biodistribution was compared with the biodistribution of other QDs in mice and rats to determine the model's predictive ability across QD types, species, and exposure routes. The model predicted the experimentally observed persistence of QDs in tissues but not early time profiles or different QD biodistribution. Therefore, more complex models will be needed to better predict QD biodistribution in vivo. PMID- 19128006 TI - Remote optical switch for localized and selective control of gene interference. AB - Near infrared-absorbing gold nanoplasmonic particles (GNPs) are used as optical switches of gene interference and are remotely controlled using light. We have tuned optical switches to a wavelength where cellular photodamage is minimized. Optical switches are functionalized with double-stranded oligonucleotides. At desired times and at specific intracellular locations, remote optical excitation is used to liberate gene-interfering oligonucleotides. We demonstrate a novel gene-interfering technique offering spatial and temporal control, which is otherwise impossible using conventional gene-interfering techniques. PMID- 19128007 TI - Stabilization of beta-galactosidase (from peas) by immobilization onto amberlite MB-150 beads and its application in lactose hydrolysis. AB - The soluble PsBGAL (from Pisum sativum ) is extremely unstable with loss of over 80% in enzyme activity within 24 h at 4 degrees C when the protein concentration was lower than 0.1 mg/mL. Enzyme immobilization onto Amberlite MB-150 beads (diameter = 5 microm) greatly stabilized the enzyme preparation, with almost no loss for 12 months at room temperature (27 degrees C). Enzyme (21.9 microg) was immobilized by 62.56% onto activated 100 mg of Amberlite MB-150 beads using 4% glutaraldehyde, at pH 6.0 (50 mM, sodium phosphate buffer). Statistical analysis carried out by ANOVA revealed that all parameters used during immobilization were equally important at P < 0.05 (level of significance). An approach toward commercial exploitation of Amberlite-PsBGAL especially in lactose hydrolysis was anticipated due to improved physicochemical properties including broad optimum pH and temperature, with a K(m) of 4.11 +/- 0.21 mM for lactose. Amberlite-PsBGAL hydrolyzed 64.57 and 69.18% of lactose present in milk and milk whey, respectively, within 10 h (at room temperature). Immobilized enzyme has reusability of over 10 batchwise uses, with almost no loss in activity. The easy accessibility of enzyme source, ease of its immobilization on Amberlite, lower cost of Amberlite, enhanced stability of Amberlite-PsBGAL, and comparable lactose hydrolysis in milk and milk whey described here make it a suitable product for future applications at laboratory and industrial scale. PMID- 19128008 TI - Er(3+)-doped nanoparticles for optical detection of magnetic field. AB - A bright persistent photoluminescence has been observed in Er(3+)-doped nanoparticles prepared by selective dissolution of bulk oxyfluoride nano-glass ceramics. A 2 orders of magnitude decrease of intensity of the (4)S(3/2)- >(4)I(15/2) green emission band of Er(3+) in these nanoparticles is observed in magnetic fields up to 50 T. This strong luminescence sensitivity to magnetic field can be used for localization and distant optical detection of magnetic field in nanovolumes with a field-resolution of 0.01 T. PMID- 19128009 TI - Association between polymerization degree of apple peel polyphenols and inhibition of Helicobacter pylori urease. AB - Apple peel extracts and their fractions pooled according to their molecular size were prepared and evaluated for their inhibitory activity against Helicobacter pylori and Jack bean ureases. Urease Inhibitory effect of apple peel polyphenols (APPE) extracted from the Granny Smith variety was concentration-dependent and reversible. High molecular weight polyphenols (HMW) were more active against Helicobacter pylori and Jack bean ureases than low molecular weight polyphenols with IC50 values of 119 and 800 microg GAE/mL, respectively. The results suggest that monomeric compounds (mainly flavan-3-ols-and quercetin-O-glycosides) will not be implicated in the antiurease effect displayed by the apple peel polyphenolic extract. Thus, as a byproduct, apple peel is suitable for developing functional ingredients that could be useful for neutralizing an important Helicobacter pylori colonization factor. PMID- 19128010 TI - Soil bacteria elevate essential oil accumulation and emissions in sweet basil. AB - Plant growth-promoting rhizobacteria ameliorate environmental conditions for plants by facilitating nutrient uptake and mitigating disease susceptibility. While volatile chemicals from certain soil microbes are sufficient to elicit growth and defense responses in Arabidopsis, whether such volatile signals can induce essential oil accumulation and chemical emissions has yet to be reported. Here, we provide biochemical evidence that the plant growth-promoting soil bacterium Bacillus subtilis GB03 releases volatile chemicals that elevate fresh weight essential oil accumulation and emissions along with plant size in the terpene-rich herb sweet basil (Ocimum basilicum). The two major essential oil components from sweet basil, alpha-terpineol and eugenol, increased ca. 2- and 10 fold, respectively, in plants exposed to GB03 volatiles or with root inoculation as compared to water controls. On a fresh and dry weight basis, shoot and root biomass increases of ca. 2-fold were observed with GB03 volatile exposure or GB03 media inoculation as compared with controls. In testing the efficacy of GB03 volatiles to trigger plant growth and secondary compound production, a physical partition separating roots from bacterial media was provided to preclude nonvolatile microbial elicitors from contributing to GB03-stimulated basil responses. These results demonstrate that volatile bacterial elicitors can concomitantly increase essential oil production and biomass in an herbaceous species rich in commercially valued essential oils. PMID- 19128011 TI - Amides and benzenoids from Zanthoxylum ailanthoides with inhibitory activity on superoxide generation and elastase release by neutrophils. AB - Five new compounds, ailanthamide (1), N-(4-methoxyphenethyl)-N-methylbenzamide (2), (2E,4E)-N-isobutyl-6-oxohepta-2,4-dienamide (3), 4-(4'-hydroxy-3' methylbutoxy)benzaldehyde (4), and (E)-methyl 4-[4-(3-hydroxypropyl)phenoxy]-2 methylbut-2-enoate (5), and 17 known compounds have been isolated from the stem bark of Zanthoxylum ailanthoides. The structures were determined through spectroscopic and MS analyses. Compounds 1, 3, xanthyletin, decarine, (+) episesamin, (-)-hinokinin, and evofolin-B exhibited inhibition (IC(50) < or = 5.34 microg/mL) of superoxide anion generation by human neutrophils in response to formyl-l-methionyl-l-leucyl-l-phenylalanine/cytochalasin B (fMLP/CB). Compounds 1, xanthyletin, decarine, and (+)-episesamin also inhibited fMLP/CB induced elastase release with IC(50) values < or = 5.53 microg/mL. PMID- 19128012 TI - In vitro binding of heavy metals by an edible biopolymer poly(gamma-glutamic acid). AB - An edible biopolymer poly(gamma-glutamic acid) (gamma-PGA) was evaluated for possible use as an chelating/binding agent in the treatment of metal intoxication in humans. In vitro binding of the toxic heavy metals lead and cadmium as affected by pH, contact time, metal concentration, gamma-PGA dose, and essential metals was carried out in a batch mode. A maximum binding occurred in the pH range 5-7, corresponding to the gastrointestinal pH values except for the stomach. Binding isotherms at pH 5.5 were well described by the heterogeneous models (Freundlich and Toth), while the lead isotherm at pH 2.5 showed a S-type curve, which was fitted as multiple curves with the Langmuir model and a shifted squared Langmuir model. However, no adsorption occurred for cadmium at pH 2.5. The maximum binding capacities of lead and cadmium at pH 5.5 were 213.58 and 41.85 mg/g, respectively. A curvilinear biphasic Scatchard plot signified a multisite interaction of metals. Binding was extremely rapid with 70-100% of total adsorption being attained in 2 min. Kinetics at low and high metal concentrations obeyed pseudo-first-order and pseudo-second-order models, respectively. The gamma-PGA dose-activity relationship revealed a low dose of gamma-PGA to be more efficient in binding a large amount of metals. Incorporation of Cu, Zn, Fe, Mg, Ca, and K showed only a minor influence on lead binding but significantly reduced the binding of cadmium. PMID- 19128013 TI - Sweetening glycoprotein processing. PMID- 19128014 TI - Evaluation of archival time on shotgun proteomics of formalin-fixed and paraffin embedded tissues. AB - There is increasing acceptance of the critical importance of correlating the morphologic features of tissue with the data obtained from various molecular analytic techniques. Access to archived formalin-fixed and paraffin-embedded (FFPE) tissue specimens via shotgun-based proteomic analyses may, therefore, open new avenues for both prospective and retrospective translational research. However, one of the remaining issues in performing comparative proteomic measurements among FFPE tissues relates to potential variability in protein composition and retrieval based on length of storage periods. Optimized protein extraction and digestion procedures for handling FFPE tissues are coupled with the capillary isotachophoresis-based proteome technology to evaluate the effects of length of storage period on archival tissue proteome analysis across 10 archived uterine mesenchymal tumor tissue blocks, including 9 uterine leiomyomas dating from 1990 to 2002 and a single case of alveolar soft part sarcoma (ASPS) from 1980. Several statistical measures, including the Pearson correlation coefficient, coefficient of variance, k-means clustering, and ANOVA, are employed to evaluate the possibility of an archival effect on individual proteins or groups of proteins within nine leiomyomas. Low abundance proteins may be more susceptible to the long-term storage as these proteins are more difficult to be retrieved and extracted as the tissue block ages in paraffin. Despite using tissue blocks stored for as many as 28 years, high confidence and comparative proteome analysis between the leiomyomas and the sarcoma is achieved. Though sharing over 1800 common proteins in a core set, a total of 80 proteins unique to the sarcoma are identified distinguishing the ASPS from the leiomyomas. Vacuolar proton translocating ATPase 116 kDa subunit isoform a3, one of the unique proteins expressed in the ASPS, is further validated by immunohistochemistry (IHC). Although IHC is highly sensitive and provides the subcellular resolution, mass spectrometry-based proteome profiling enables global identification and quantification of thousands of proteins without a priori knowledge of individual proteins being analyzed or the need of validated antibodies. PMID- 19128015 TI - Structures of falcipain-2 and falcipain-3 bound to small molecule inhibitors: implications for substrate specificity. AB - Falcipain-2 and falcipain-3 are critical hemoglobinases of Plasmodium falciparum, the most virulent human malaria parasite. We have determined the 2.9 A crystal structure of falcipain-2 in complex with the epoxysuccinate E64 and the 2.5 A crystal structure of falcipain-3 in complex with the aldehyde leupeptin. These complexes represent the first crystal structures of plasmodial cysteine proteases with small molecule inhibitors and the first reported crystal structure of falcipain-3. Our structural analyses indicate that the relative shape and flexibility of the S2 pocket are affected by a number of discrete amino acid substitutions. The cumulative effect of subtle differences, including those at "gatekeeper" positions, may explain the observed kinetic differences between these two closely related enzymes. PMID- 19128016 TI - Structural evolutions of salicylaldoximes as selective agonists for estrogen receptor beta. AB - The bioisosteric replacement of the phenol ring, a signature functional group of most estrogen receptor (ER) ligands, with a hydrogen-bonded pseudocyclic ring, led to the development of a novel class of nonsteroidal ER-ligands based on a salicylaldoxime template. A series of structural modifications were applied to selected molecules belonging to the monoaryl-salicylaldoxime chemical class in an attempt to improve further their ERbeta-selective receptor affinity and agonist properties. Among several modifications, the best results were obtained by the simultaneous introduction of a meta-fluorine atom into the para-hydroxyphenyl substituent present in the 4-position of salicylaldoxime, together with the insertion of a chloro group in the 3-position of the central scaffold. The resulting compound showed the best affinity (K(i) = 7.1 nM) and selectivity for ERbeta over ERalpha. Moreover, in transcription assays, it proved to be a selective and potent ERbeta-full agonist with an EC(50) of 4.8 nM. PMID- 19128017 TI - Plasma induced grafting carboxymethyl cellulose on multiwalled carbon nanotubes for the removal of UO(2)(2+) from aqueous solution. AB - Carboxymethyl cellulose (CMC) is grafted on multiwalled carbon nanotubes (MWCNT) by using plasma techniques. The CMC grafted MWCNT (MWCNT-g-CMC) is characterized by using Fourier transform infrared spectra (FT-IR), Raman spectra, powder X-ray diffraction (XRD), thermogravimetric analysis (TGA)-differential thermal analysis (DTA), scanning electron microscopy (SEM), and N(2)-BET methods in detail. The application of MWCNT-g-CMC in the removal of UO(2)(2+) from aqueous solution is investigated. MWCNT-g-CMC has much higher sorption ability in the removal of UO(2)(2+) than raw MWCNT. The MWCNT-g-CMC is a suitable material in the preconcentration and solidification of heavy metal ions from large volume of aqueous solutions. PMID- 19128018 TI - Conformational transitions of immobilized DNA chains driven by pH with electrochemical output. AB - A pH-driven DNA sway rod is prepared by immobilizing thiolated DNA, mercaptohexanol, and cysteine on a gold electrode surface. As pH changes around the pI of cysteine, contrary electrostatic effect is produced between the negative DNA and amphoteric cysteine, which actuates reversible conformational transitions, such as sway of the DNA molecules, rodlike chain-to-globule, and so forth. The nanoscale motion can be detected by commonly used electrochemical technique and reversible electrochemical signal may be observed. PMID- 19128019 TI - Structure and dynamics of the ApA, ApC, CpA, and CpC RNA dinucleoside monophosphates resolved with NMR scalar spin-spin couplings. AB - The measured NMR scalar coupling constants (J-couplings) in the XpY, (X,Y = adenine (A) or cytosine (C)) RNA dinucleoside monophosphates (DMPs) were assigned to the backbone (alpha, beta, gamma, delta, epsilon, zeta) and glycosidic (chi) torsion angles in order to resolve the global structure of the DMP molecules. The experimental J-couplings were correlated with the theoretical J-couplings obtained as the dynamical averages of the Karplus equations relevant to the torsion angles. The dynamical information was captured using the molecular dynamics (MD) calculation method. The individual conformational flexibility of the four DMP molecules was thus consistently probed with the NMR J-couplings. The calculated structure and flexibility of the DMP molecules depend on the sequence considered with respect to the 5' and 3' end of the DMP molecules (5'-XpY-3'). The dynamical characteristics of the two nucleosides are not equivalent even for the ApA and CpC homologues. An enhancement of the sampling in the MD calculations was achieved using five different starting structural motives classified previously for the RNA backbone in the solid phase (Richardson et al. RNA 2008, 14, 465-481). The initial structures were selected on the basis of a database search for RNA oligonucleotides. Frequent interconversions between the conformers during the MD calculations were actually observed. The structural interpretation of the NMR spectroscopic data based on the MD simulations combined with the Karplus equations indicates that the dominant conformation of the DMP molecules in solution corresponds to the A-RNA form. For 52% of the total simulation time (1000 ns), the zeta(g-)-alpha(g-)-gamma(g+) backbone topology corresponding to the canonical A-RNA form was observed, with roughly equally populated C2'- and C3'-endo sugar puckers interconverting on the nanosecond time scale. However, other noncanonical patterns were also found and thus indicate their relatively high potential to be populated in the dynamical regime. For approximately 72% of the time portion when the A-RNA of the zeta-alpha-gamma combination occurred, the nucleobases were classified as being mutually stacked. The geometries of the nucleobases classified in this work as stacked were significantly more populated for the DMP molecules with adenosine at the 3' end (ApA and CpA DMPs) than the ApC or CpC RNA molecules with C at the 3' end. PMID- 19128020 TI - Integral absorption coefficients of C-H stretching bands in IR spectra of ethane adsorbed by cationic forms of Y zeolite. AB - An attempt to measure absolute intensities of IR C-H stretching bands of ethane specifically adsorbed by Na+, Ca2+, or Mg2+ cations in Y zeolite was carried out via simultaneous transmission IR and volumetric measurements at very low pressures using self-supporting wafers. For wafers with a thickness of 7-9 mg/cm2 quantitative measurements of absorption coefficients of C-H stretching bands were possible. More pronounced shifts of C-H bands toward lower frequencies indicated that the perturbation of the corresponding bonds in ethane upon adsorption by the bivalent Ca2+ or Mg2+ ions was much stronger than by the monovalent Na+ ions. In addition, the integral extinction coefficient of the C-H stretching bands of ethane adsorbed by Na+ ions was only about 53% of that of gas-phase ethane. For adsorption on the bivalent Ca2+ or Mg2+ ions the coefficients exceeded those of the free ethane molecule, indicating strong polarization of the C-H bonds during the vibration. Bands representing the fully symmetric C-H stretching vibrations, which are IR inactive for free ethane, were perturbed most strongly upon adsorption. Their intensities reached up to 60% of the total intensity in the C-H stretching region. In contrast, the intensities of the other C-H bands were several times lower than for free ethane. These trends indicate anisotropy in the polarizability of the adsorbed molecules. In addition, the absorption coefficients indicate a weaker adsorption and polarization of ethane by Mg2+ than by Ca2+ ions in Y zeolite. Because of their smaller ionic radius, the Mg2+ cations are more strongly shielded by the surrounding oxygen atoms of the zeolite framework than the Ca2+ cations, and therefore, their polarizing ability is decreased. PMID- 19128021 TI - Computational methods in organic thermochemistry. 4. Enthalpies and Gibbs energies of formation of the cis- and trans-diazenes. AB - The enthalpies and Gibbs energies of formation are examined for over 100 aliphatic cis- and trans-diazenes using the G3, G3(MP2), and CBS-QB3 model chemistries for compounds ranging from two to ten heavy atoms. The values computed using bond separation reactions were corrected to compensate for the presence of higher energy conformers. Comparison of several homologous and isomeric series shows that alkene thermochemistry is not an adequate model for the diazenes (HN=NH) as the group equivalents of the latter differ significantly from those of the former when they are adjacent or one atom removed from the nitrogen-nitrogen double bond. Methylene insertion adjacent to the double bond is less exothermic for the diazenes than it is for the alkenes whereas the same insertion one carbon atom away from the double bond is more exothermic than that observed for the alkenes. The data argues that molecular modeling programs parametrized using existing experimental data will need to be recalibrated. PMID- 19128022 TI - Selective paraffin removal from ethane/ethylene mixtures by adsorption into aluminum methylphosphonate-alpha: a molecular simulation study. AB - Most adsorbent materials used for olefin/paraffin separation show preferential adsorption of the olefin. Recently, the material aluminum methylphosphonate polymorph alpha (AlMePO-alpha) was found to be able to selectively adsorb the paraffin instead of the olefin, from an ethyl chloride/vinyl chloride mixture (Herdes, C.; Valente, A.; Lin, Z.; Rocha, J.; Coutinho, J. A. P.; Medina, F.; Vega, L. F. Langmuir 2007, 23, 7299). However, several questions remain still open regarding the reasons for this selective paraffin adsorption, as well as the suitability of AlMePO-alpha as adsorbent for other olefin/paraffin separations. In this work, the adsorption of ethane/ethylene mixtures by AlMePO-alpha is investigated using grand canonical Monte Carlo simulations in order to determine the effect of molecular interactions, size, and shape on the selective adsorption. For this purpose three different force fields have been used for the fluids, investigating the effect of the molecular details of the fluid on the adsorption behavior. All three force fields gave the same qualitative behavior. It was found that AlMePO-alpha is also able to selectively adsorb the paraffin from ethane/ethylene mixtures. Moreover, ethane molecules arrange exactly in the same way in the adsorbent material as ethyl chloride, with the methyl groups directed toward each other, although ethane has much smaller dynamic diameter compared to ethyl chloride. Therefore, a key factor determining the selectivity is found to be the molecular interaction between the methyl group of AlMePO-alpha and the methyl group of the paraffin. PMID- 19128023 TI - Immobilization of penicillin G acylase on oxirane-modified mesoporous silicas. AB - Experimental adsorption kinetics and equilibrium results of penicillin G acylase (PGA, from Escherichia coli, EC 3.5.1.11) on mesoporous silicas with pore sizes ranging from 5.6 to 33.2 nm showed that samples with pore sizes between 11.0 and 13.2 nm exhibited the best performance in immobilizing PGA under mild experimental conditions. A mesoporous silica sample with an optimum pore size of about 11.5 nm was then modified with different amounts of glycidoxypropyltrimethoxysilane to yield oxirane-functionalized silicas of different densities of surface oxirane groups. Under very mild incubation conditions, a partially oxirane-functionalized silica sample was found to be more efficient in immobilizing PGA than a fully oxirane-functionalized sample and a commercial polymer carrier (i.e., Eupergit C). With the partially oxirane functionalized mesoporous silica sample as a carrier, a PGA loading of 110 mg/g (dry support) and an enzymatic activity of as high as 3477 unit/g (dry support) were achieved within 24 h of incubation. The residual surface silanol groups on the partially oxirane-functionalized silica were observed to play a pivotal role in facilitating the covalent binding of PGA with the oxirane groups at low salt concentrations. PMID- 19128024 TI - Tetragonal to triclinic--a phase change for [Fe(TPP)(NO)]. AB - The temperature dependence of the crystalline phase of (nitrosyl)(tetraphenylporphinato)iron(II), [Fe(TPP)(NO)], has been explored over the temperature range of 33-293 K. The crystalline complex is found in the tetragonal crystal system at higher temperatures and in the triclinic crystal system at lower temperatures. In the tetragonal system, the axial ligand is strongly disordered, with the molecule having crystallographically required 4/m symmetry, leading to eight distinct positions of the single nitrosyl oxygen atom. The phase transition to the triclinic crystal system leads to a partial ordering with the molecule now having inversion symmetry and disorder of the axial nitrosyl ligand over only two positions. The increase in ordering allows subtle molecular geometry features to be observed; in particular, an off-axis tilt of the Fe-N(NO) bond from the heme normal is apparent. The transition of the reversible phase change begins at about 250 K. This transition has been confirmed by both X-ray diffraction studies and a differential scanning calorimetry study. PMID- 19128025 TI - Enantioselective total synthesis of aspergillide C. AB - The first enantioselective total synthesis of aspergillide C, a cytotoxic 14 membered macrolide isolated from the marine-derived fungus Aspergillus ostianus, has been accomplished from a commercially available chiral glycidol derivative by a 12-step sequence involving an expeditious preparation of a cyclic acetal intermediate and a trans-selective Ferrier-type two-carbon homologation reaction. PMID- 19128026 TI - Architectural underpinnings of the genetic code for glutamine. AB - Structure-based mutational analysis was used to probe the architecture of the glutamine binding pocket in Escherichia coli glutaminyl-tRNA synthetase (GlnRS). Crystallographic studies of several different GlnRS complexes in a lattice that supports catalytic activity have shown that the glutamine amide group makes only ambiguous hydrogen-bonding interactions with a tyrosine hydroxyl and bound water molecule, rather than the highly specific hydrogen-bonding and electrostatic interactions made by the substrate amino acid in all other nonediting tRNA synthetases. Further, the amide oxygen of substrate glutamine accepts a hydrogen bond from the 3'-ribose hydroxyl group of ATP, an unusual distal substrate substrate interaction also not observed in any other tRNA synthetase complex. Steady-state and pre-steady-state kinetic analysis using a 3'-dATP analogue in place of ATP shows that removal of this distal interaction does not affect K(m) for the analogue as compared with ATP, yet decreases the efficiency of aminoacylation by 10(3)-fold while significantly elevating K(m) for glutamine. In other experiments, mutation of eight nearly fully conserved residues in the glutamine binding pocket reveals decreases in k(cat)/K(m) ranging from 5- to 400 fold, and in K(d) for glutamine of up to at least 60-fold. Amino acid replacements at Tyr211 and Gln255, which participate with substrate glutamine in an antidromic circular arrangement of hydrogen bonds, cause the most severe decreases in catalytic efficiency. This finding suggests that the relative absence of direct hydrogen bonds to glutamine may be in part compensated by additional binding energy derived from the enhanced stability of this circular network. Calculations of electrostatic surface potential in the active site further suggest that a complementary electrostatic environment is also an important determinant of glutamine binding. PMID- 19128027 TI - A new strategy for the synthesis of optically pure beta-fluoroalkyl beta-amino acid derivatives. AB - The first general approach for the diastereoselective formation of a variety of optically pure anti-beta-fluoroalkyl beta-amino acid derivatives is described. The process relies on the stereocontrolled reaction, mediated by a remote sulfoxide, of fluorinated imines with sulfinylated benzyl carbanions, which are used as synthetic equivalents of chiral ester enolates. PMID- 19128028 TI - Clogging the periplasmic pathway in LacY. AB - The lactose permease of Escherichia coli (LacY) is a highly dynamic membrane transport protein. Crystal structures of wild-type and mutant LacY all exhibit an inward-facing conformation with an open cytoplasmic pathway and a tightly packed periplasmic side, which makes the binding site inaccessible from the outside. However, biochemical and biophysical findings provide strong evidence that occupation of the sugar-binding site leads to an increased probability of opening of a hydrophilic pathway on the periplasmic side and closing of the cytoplasmic cavity. By this means, the sugar-binding site becomes accessible to either side of the membrane in alternating fashion. To extend studies on the relationship between the periplasmic pathway and transport activity, engineered single-Cys replacements in the periplasmic pathway were reacted to completion with thiol reagents, and the effects on transport and sugar binding were tested. Inactivation correlates for the most part with the size of the modifying reagent, although the position of the Cys replacement is also important. However, sugar binding is unaffected. The results suggest that placement of a relatively large moiety in the putative periplasmic cleft of LacY likely prevents closure, an essential step in the transport cycle, without significantly altering access of sugar to the binding site. PMID- 19128029 TI - Critical role of the solvent environment in galectin-1 binding to the disaccharide lactose. AB - Galectin-1 (Gal-1), a member of a family of evolutionarily conserved glycan binding proteins, binds specifically to poly-N-acetyllactosamine-enriched glycoconjugates. Through interactions with these glycoconjugates, this protein modulates inflammatory responses and contributes to tumor progression and immune cell homeostasis. The carbohydrate recognition domain includes the single protein tryptophan (Trp68). UV resonance Raman spectroscopy and molecular dynamic simulation were used to examine the change in the environment of the Trp on ligand binding. The UV Raman spectra and the calculated water radial distribution functions show that, while no large structural changes in the protein follow lactose binding, substantial solvent reorganization occurs. These new insights into the microscopic role of water molecules in Gal-1 binding to its specific carbohydrate ligands provides a better understanding of the physicochemical properties of Gal-1-saccharide interactions, which will be useful for the design of synthetic inhibitors for therapeutic purposes. PMID- 19128031 TI - Concurrent binding of complexin and synaptotagmin to liposome-embedded SNARE complexes. AB - Synaptotagmin and complexin regulate SNARE-mediated synaptic vesicle exocytosis. It has been proposed that complexin clamps membrane fusion and that Ca(2+) synaptotagmin displaces complexin from SNARE complexes to relieve this clamping activity. Using a reconstituted system, we demonstrate that complexin and synaptotagmin simultaneously bind to neuronal SNARE complexes and that both apo synaptotagmin and complexin inhibit SNARE-mediated membrane fusion. Moreover, the clamping ability of apo-synaptotagmin occluded the clamping activity of complexin until the arrival of a Ca(2+) trigger, at which point synaptotagmin accelerated fusion while high concentrations of complexin inhibited fusion. Thus, the inhibitory patterns of synaptotagmin and complexin are different, suggesting that SNAREs assemble into distinct states along the fusion pathway. These data also suggest that during synaptotagmin-regulated vesicle-vesicle fusion, complexin does not function as a fusion clamp that is relieved by Ca(2+)-synaptotagmin. PMID- 19128032 TI - Trypanosoma cruzi induces life-history trait changes in the wild kissing bug Mepraia spinolai: implications for parasite transmission. AB - One important paradigm in host-parasite evolutionary biology is the ability of parasites to manipulate the phenotype of their hosts to facilitate transmission. In this paper, I examine whether the protozoan parasite Trypanosoma cruzi modifies the developmental time, body size, and survival of its vector, the bloodsucking insect Mepraia spinolai (Hemiptera; Reduviidae). M. spinolai nymphs were experimentally infected when fed on T. cruzi-infected mice (infected group) or kept uninfected when fed on healthy mice (control group). T. cruzi-infected insects showed a retarded developmental time and reduced survival compared with uninfected individuals. The impact of the parasite on the vector was age dependent as the last three insect molts were the most affected stages. The presence of T. cruzi decreased significantly the weight of male and female insects in the three last stages. When insect sex was taken into account, infected female bugs took longer than infected males to develop into the adult stage, which implies that the impact of T. cruzi is sex-dependent. Results from this study indicate that T. cruzi has a strong impact on life history traits of M. spinolai and provide strong evidence of age- and sex-dependent parasite induced phenotype modification for insect vectors. The implications of this study along with previously reported feeding behavioral alterations in this insect vector-parasite system suggest that T. cruzi-induced modifications could translate into an enhanced transmission to definitive mammal hosts. PMID- 19128030 TI - Dihydroorotase from the hyperthermophile Aquifex aeolicus is activated by stoichiometric association with aspartate transcarbamoylase and forms a one-pot reactor for pyrimidine biosynthesis. AB - In prokaryotes, the first three enzymes in pyrimidine biosynthesis, carbamoyl phosphate synthetase (CPS), aspartate transcarbamoylase (ATC), and dihydroorotase (DHO), are commonly expressed separately and either function independently (Escherichia coli) or associate into multifunctional complexes (Aquifex aeolicus). In mammals the enzymes are expressed as a single polypeptide chain (CAD) in the order CPS-DHO-ATC and associate into a hexamer. This study presents the three-dimensional structure of the noncovalent hexamer of DHO and ATC from the hyperthermophile A. aeolicus at 2.3 A resolution. It is the first structure of any multienzyme complex in pyrimidine biosynthesis and is a possible model for the core of mammalian CAD. The structure has citrate, a near isosteric analogue of carbamoyl aspartate, bound to the active sites of both enzymes. Three active site loops that are intrinsically disordered in the free, inactive DHO are ordered in the complex. The reorganization also changes the peptide bond between Asp153, a ligand of the single zinc atom in DHO, and Gly154, to the rare cis conformation. In the crystal structure, six DHO and six ATC chains form a hollow dodecamer, in which the 12 active sites face an internal reaction chamber that is approximately 60 A in diameter and connected to the cytosol by narrow tunnels. The entrances and the interior of the chamber are both electropositive, which suggests that the architecture of this nanoreactor modifies the kinetics of the bisynthase, not only by steric channeling but also by preferential escape of the product, dihydroorotase, which is less negatively charged than its precursors, carbamoyl phosphate, aspartate, or carbamoyl aspartate. PMID- 19128033 TI - Mechanism of interferon-gamma-induced increase in T84 intestinal epithelial tight junction. AB - Interferon-gamma (IFN-gamma) is an important proinflammatory cytokine that plays a central role in the intestinal inflammatory process of inflammatory bowel disease. IFN-gamma induced disturbance of the intestinal epithelial tight junction (TJ) barrier has been postulated to be an important mechanism contributing to intestinal inflammation. The intracellular mechanisms that mediate the IFN-gamma induced increase in intestinal TJ permeability remain unclear. The aim of this study was to examine the role of the phosphatidylinositol 3-kinase (PI3-K) pathway in the regulation of the IFN-gamma induced increase in intestinal TJ permeability using the T84 intestinal epithelial cell line. IFN-gamma caused an increase in T84 intestinal epithelial TJ permeability and depletion of TJ protein, occludin. The IFN-gamma induced increase in TJ permeability and alteration in occludin protein was associated with rapid activation of PI3-K; and inhibition of PI3-K activation prevented the IFN-gamma induced effects. IFN-gamma also caused a delayed but more prolonged activation of nuclear factor-kappaB (NF-kappaB); inhibition of NF-kappaB also prevented the increase in T84 TJ permeability and alteration in occludin expression. The IFN-gamma induced activation of NF-kappaB was mediated by a cross talk with PI3-K pathway. In conclusion, the IFN-gamma induced increase in T84 TJ permeability and alteration in occludin protein expression were mediated by the PI3-K pathway. These results show for the first time that the IFN-gamma modulation of TJ protein and TJ barrier function is regulated by a cross-talk between PI3-K and NF-kappaB pathways. PMID- 19128035 TI - Study on the formation of self-assembled monolayers on sol-gel processed hafnium oxide as dielectric layers. AB - High dielectric constant (k) metal oxides such as hafnium oxide (HfO2) have gained significant interest due to their applications in microelectronics. In order to study and control the surface properties of hafnium oxide, self assembled monolayers (SAMs) of four different long aliphatic molecules with binding groups of phosphonic acid, carboxylic acid, and catechol were formed and characterized. Surface modification was performed to improve the interface between metal oxide and top deposited materials as well as to create suitable dielectric properties, that is, leakage current and capacitance densities, which are important in organic thin film transistors. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, contact angle goniometry, atomic force microscopy (AFM), and simple metal-HfO2-SAM-metal devices were used to characterize the surfaces before and after SAM modification on sol-gel processed hafnium oxide. The alkylphosphonic acid provided the best monolayer formation on sol-gel processed hafnium oxide to generate a well-packed, ultrathin dielectric exhibiting a low leakage current density of 2x10(-8) A/cm2 at an applied voltage of -2.0 V and high capacitance density of 0.55 microF/cm2 at 10 kHz. Dialkylcatechol showed similar characteristics and the potential for using the catechol SAMs to modify HfO2 surfaces. In addition, the integration of this alkylphosphonic acid SAM/hafnium oxide hybrid dielectric into pentacene-based thin film transistors yields low-voltage operation within 1.5 V and improved performance over bare hafnium oxide. PMID- 19128036 TI - Characterization of the inhibitor binding site in mitochondrial NADH-ubiquinone oxidoreductase by photoaffinity labeling using a quinazoline-type inhibitor. AB - The diverse inhibitors of bovine heart mitochondrial complex I (NADH-ubiquinone oxidoreductase) are believed to share a common large binding domain with partially overlapping sites, though it remains unclear how these binding sites relate to each other. To obtain new insight into the inhibitor binding domain in complex I, we synthesized a photoreactive azidoquinazoline {[(125)I]-6-azido-4-(4 iodophenethylamino)quinazoline, [(125)I]AzQ}, in which a photolabile azido group was introduced into the toxophoric quinazoline ring to allow specific cross linking, and carried out a photoaffinity labeling study using bovine heart submitochondrial particles. Analysis of the photo-cross-linked proteins by peptide mass fingerprinting and immunoblotting revealed that [(125)I]AzQ specifically binds to the 49 kDa and ND1 subunits with a frequency of approximately 4:1. The cross-linking was completely blocked by excess amounts of other inhibitors such as acetogenin and fenpyroximate. Considerable cross-linking was also detected in the ADP/ATP carrier and 3-hydroxybutyrate dehydrogenase, though it was not associated with dysfunction of the two proteins. The partial proteolysis of the [(125)I]AzQ-labeled 49 kDa subunit by V8-protease and N terminal sequencing of the resulting peptides revealed that the amino acid residue cross-linked by [(125)I]AzQ is within the sequence region Thr25-Glu143 (118 amino acids). Furthermore, examination of fragment patterns generated by exhaustive digestion of the [(125)I]AzQ-labeled 49 kDa subunit by V8-protease, lysylendopeptidase, or trypsin strongly suggested that the cross-linked residue is located within the region Asp41-Arg63 (23 amino acids). The present study has revealed, for the first time, the inhibitor binding site in complex I at the sub subunit level. PMID- 19128037 TI - Common mitochondrial DNA mutations generated through DNA-mediated charge transport. AB - Mutation sites that arise in human mitochondrial DNA as a result of oxidation by a rhodium photooxidant have been identified. HeLa cells were incubated with [Rh(phi)(2)bpy]Cl(3) (phi is 9,10-phenanthrenequinone diimine), an intercalating photooxidant, to allow the complex to enter the cell and bind mitochondrial DNA. Photoexcitation of DNA-bound [Rh(phi)(2)bpy](3+) can promote the oxidation of guanine from a distance through DNA-mediated charge transport. After two rounds of photolysis and growth of cells incubated with the rhodium complex, DNA mutations in a portion of the mitochondrial genome were assessed via manual sequencing. The mutational pattern is consistent with dG to dT transversions in the repetitive guanine tracts. Significantly, the mutational pattern found overlaps oxidative damage hot spots seen previously. These mutations are found within conserved sequence block II, a critical regulatory element involved in DNA replication, and these have been identified as sites of low oxidation potential to which oxidative damage is funneled. On the basis of this mutational analysis and its correspondence to sites of long-range oxidative damage, we infer a critical role for DNA charge transport in generating these mutations and, thus, in regulating mitochondrial DNA replication under oxidative stress. PMID- 19128038 TI - Development of a method for the genetic identification of commercial bivalve species based on mitochondrial 18S rRNA sequences. AB - In this study a genetic methodology based on the amplification of an 18S rRNA fragment by PCR and phylogenetic analysis of the obtained DNA sequences was developed. This technique allows the genetic identification of more than 50 bivalve species in fresh, frozen, precooked and canned products. The developed method was applied to 30 commercial samples to check their labeling, showing that 12 samples were incorrectly labeled (40%). Therefore, the proposed methodology is appropriate to study questions related to the correct labeling and traceability of commercial products and the control of imported bivalves and fisheries in order to guarantee the protection of consumers' rights and verify the transparency of the extractive and transforming industries. PMID- 19128039 TI - Star-shaped cationic polymers by atom transfer radical polymerization from beta cyclodextrin cores for nonviral gene delivery. AB - Cationic polymers with low cytotoxicity and high transfection efficiency have attracted considerable attention as nonviral carriers for gene delivery. Herein, well-defined and star-shaped CDPD consisting of beta-CD cores and P(DMAEMA) arms, and CDPDPE consisting of CDPD and P(PEGEEMA) end blocks (where CD = cyclodextrin, P(DMAEMA) = poly(2-(dimethylamino)ethyl methacrylate), P(PEGEEMA) = poly(poly(ethylene glycol)ethyl ether methacrylate)) for gene delivery were prepared via atom transfer radical polymerization (ATRP) from the bromoisobutyryl terminated beta-CD core. The CDPD and CDPDPE exhibit good ability to condense plasmid DNA (pDNA) into 100-200 nm size nanoparticles with positive zeta potentials of 25-40 mV at nitrogen/phosphate (N/P) ratios of 10 or higher. CDPD and CDPDPE exhibit much lower cytotoxicity and higher gene transfection efficiency than high molecular weight P(DMAEMA) homopolymers. A comparison of the transfection efficiencies between CDPD and P(DMAEMA) homopolymer indicates that the unique star-shaped architecture involving the CD core can enhance the gene transfection efficiency. In addition to reducing cytotoxicity, the introduction of a biocompatible P(PEGEEMA) end block to the P(DMAEMA) arms in CDPDPE can further enhance the gene transfection efficiency. PMID- 19128040 TI - Phenolic antioxidants from green tea produced from Camellia crassicolumna Var. multiplex. AB - Camellia crassicolumna var. multiplex (Chang et Tan) Ming belonging to Camellia sect. Thea (Theaceae), is endemic to the southeastern area of Yunnan province, China, where the leaves have been commonly used for making tea and beverages consumed widely. HPLC analysis showed that there was no caffeine or theophylline contained in the leaves; however, thin layer chromatography (TLC) analysis suggested the abundant existence of phenolic compounds. Further detailed chemical investigation of the green tea produced from the leaves of the plant led to the identification of 18 phenolic compounds, including four flavan-3-ols (1-4), six flavonol glycosides (5-10), three hydrolyzable tannins (11-13), two chlorogenic acid derivatives (14, 15), and three simple phenolic compounds (16-18). The isolated compounds were evaluated for their antioxidant activities by 1,1' diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and tyrosinase inhibitory assays. Most of them exhibited significant DPPH radical scavenging activities, whereas flavan-3-ols and hydrolyzable tannins showed stronger inhibitory activities on tyrosinase. The results suggest that C. crassicolumna could be an ideal plant resource for a noncaffeine beverage. PMID- 19128041 TI - Effect of Prolyl-hydroxyproline (Pro-Hyp), a food-derived collagen peptide in human blood, on growth of fibroblasts from mouse skin. AB - We examined the effect of prolyl-hydroxyproline (Pro-Hyp), which occurs in human peripheral blood after ingestion of collagen peptide, on the migration and growth of mouse skin fibroblasts. Mouse skin discs were cultured on a 24-well plastic plate in a fetal bovine serum (FBS)-free medium. Addition of Pro-Hyp (200 nmol/mL) significantly increased the number of fibroblasts migrating from the skin to the plate after incubation for 72 h. This effect of Pro-Hyp was abolished by the addition of mitomycin C. The fibroblasts that had migrated from the mouse skin were collected and cultured on collagen gel. The growth of fibroblasts on the collagen gel was suppressed even in the presence of FBS, while rapid fibroblast growth was observed on the plastic plate. Addition of Pro-Hyp (0-1000 nmol/mL) to the medium containing 10% FBS enhanced the growth of fibroblasts on the collagen gel in a dose-dependent manner. These results suggest that Pro-Hyp might stimulate the growth of fibroblasts in the skin and consequently increase the number of fibroblasts migrating from the skin. PMID- 19128042 TI - Synthetic approaches to the microtubule-stabilizing sponge alkaloid ceratamine A and desbromo analogues. AB - Two synthetic approaches to the microtubule-stabilizing ceratamine alkaloids are described. The first approach involved attempts to graft an aminoimidazole moiety onto an azepine ring to form partially hydrogenated versions of the unprecedented aromatic imidazo[4,5-d]azepine core of the ceratamines. This route ultimately failed because it was not possible to aromatize the partially hydrogenated ceratamine intermediates. A second approach started with tribromoimidazole that was sequentially metalated and functionalized to efficiently generate a key imidazole intermediate containing vinyl bromide and amide functionalities. An intramolecular Buchwald vinyl amidation reaction converted this key intermediate into a bicyclic imidazo[4,5-d]azepine that was at the same oxidation state as the aromatic core of the ceratamines. The 2-amino functionality present on the imidazole ring of the ceratamines was installed using a Buchwald/Hartwig amination reaction on a 2-chloroimidazole precursor. Deprotection and aromatization resulted in the first synthesis of desbromoceratamine A (55) and desmethyldesbromoceratamine A (60). An unanticipated addition of atmospheric oxygen was encountered during deprotection of the imidazole ring in the last step of the synthesis leading to C-11 oxygenated ceratamine analogues as byproducts. Evaluation of the synthetic ceratamines in a TG3 cell-based assay for mitotic arrest revealed that the C-14 and C-16 bromine substituents in ceratamine A (1) play a major role in the antimitotic potency of the natural product. The synthetic route to ceratamine analogues has provided sufficient quantities of desbromoceratamine A (55) for testing in mouse models of cancer. PMID- 19128043 TI - Distance and orientation dependence of excitation energy transfer: from molecular systems to metal nanoparticles. AB - The elegant theory developed by Forster to describe the rate of fluorescence resonance energy transfer between a donor and an acceptor has played a key role in understanding the structure and dynamics of polymers, biopolymers (proteins, nucleic acids), and self-assemblies (photosystems, micellar systems). Forster theory assumes the transition charge densities of donor and acceptor molecules are point dipoles and hence predicts a 1/R(6) dependence of energy transfer rate on center-to-center separation distance, R. In addition, a preaveraging over the orientations of the two dipoles is usually performed. The present review examines the validity of these assumptions in following different donor-acceptor (D-A) systems: (i) dye molecules attached to a flexible polymer chain in solution, (ii) extended conjugated dye molecules in quenched conformation, (iii) dye and a spherical metal nanoparticle of different sizes, (iv) two spherical metal nanoparticles, and (v) two prolate shaped metal nanoparticles at different relative orientations. In the case of dye molecules attached to a flexible polymer chain, we discuss the recent theoretical and computer simulation studies of energy transfer dynamics. It includes an analysis of Wilemski-Fixman (WF) theory of a bimolecular reaction in solution, applied to the excitation energy transfer between two ends of the polymer. We briefly describe the limitation of the WF theory and its generalizations that lead to a better agreement between the theory and the simulation results. The orientational dynamics of dye molecules is found to significantly influence the rate of excitation energy transfer, and may play a "hidden role" in influencing the observed distance dependence. For extended conjugated D-A systems and those involving nonspherical metal nanoparticles, even at intermediate separations, a significant deviation from 1/R(6)-type distance dependence of the energy transfer rate is found. Surprisingly, however, this distance dependence is robust for D-A systems involving spherical metal nanoparticles. For both spherical and nonspherical metal nanoparticles (MNps), the functional dependence of rate on the surface-to surface separation distance (d) is quite different, at small to intermediate distances (compared to the size of the MNps). The rate calculations of excitation energy transfer between extended conjugated dye molecules reveal that optically dark states can significantly contribute toward enhancing the energy transfer rate. It is further found that the rate of energy transfer between nonspherical metal nanoparticles exhibits an interesting orientation dependence not anticipated in Forster's approach. PMID- 19128044 TI - Possible existence of convective currents in surfactant bulk solution in experimental pendant-bubble dynamic surface tension measurements. AB - Traditionally, surfactant bulk solutions in which dynamic surface tension (DST) measurements are conducted using the pendant-bubble apparatus are assumed to be quiescent. Consequently, the transport of surfactant molecules in the bulk solution is often modeled as being purely diffusive when analyzing the experimental pendant-bubble DST data. In this Article, we analyze the experimental pendant-bubble DST data of the alkyl poly (ethylene oxide) nonionic surfactants, C12E4 and C12E6, and demonstrate that both surfactants exhibit "superdiffusive" adsorption kinetics behavior with characteristics that challenge the traditional assumption of a quiescent surfactant bulk solution. In other words, the observed superdiffusive adsorption behavior points to the possible existence of convection currents in the surfactant bulk solution. The analysis presented here involves the following steps: (1) constructing an adsorption kinetics model that corresponds to the fastest rate at which surfactant molecules adsorb onto the actual pendant-bubble surface from a quiescent solution, (2) predicting the DST behaviors of C12E4 and C12E6 at several surfactant bulk solution concentrations using the model constructed in step 1, and (3) comparing the predicted DST profiles with the experimental DST profiles. This comparison reveals systematic deviations for both C12E4 and C12E6 with the following characteristics: (a) the experimental DST profiles exhibit adsorption kinetics behavior, which is faster than the predicted fastest rate of surfactant adsorption from a quiescent surfactant bulk solution at time scales greater than 100 s, and (b) the experimental DST profiles and the predicted DST behaviors approach the same equilibrium surface tension values. Characteristic (b) indicates that the cause of the observed systematic deviations may be associated with the adsorption kinetics mechanism adopted in the model used rather than with the equilibrium behavior. Characteristic (a) indicates that the actual surfactant bulk solution in which the DST measurement was conducted, most likely, cannot be considered to be quiescent at time scales greater than 100 s. Accordingly, the observed superdiffusive adsorption behavior is interpreted as resulting from convection currents present in a nonquiescent surfactant bulk solution. Convection currents accelerate the surfactant adsorption process by increasing the rate of surfactant transport in the bulk solution. The systematic nature of the deviations observed between the predicted DST profiles and the experimental DST behavior for C12E4 and C12E6 suggests that the nonquiescent nature of the surfactant bulk solution may be intrinsic to the experimental pendant-bubble DST measurement approach. To validate this possibility, we identified generic features in the experimental DST data when DST measurements are conducted in a nonquiescent surfactant bulk solution, and the DST measurements are analyzed assuming that the surfactant bulk solution is quiescent. An examination of the DST literature reveals that these identified generic features are quite general and are observed in the experimental DST data of several other surfactants (decanol, nonanol, C10E8, C14E8, C12E8, and C10E4) measured using the pendant bubble apparatus. PMID- 19128046 TI - Multicomponent assembly of heterometallic isosceles triangles. AB - The multicomponent synthesis and solution-state characterization of three supramolecular bis-heterometallic isosceles triangles are elaborated. The triangular assemblies are isosceles both geometrically and chemically; they comprise multiple ligands, metals, and binding motifs. Variation of the length of one side of the triangle by changing the number of phenyl spacers n = 0, 1, and 2 influences the redox potential of the opposing copper(I) center, allowing translation of the nanomechanical changes into electronically readable values. PMID- 19128047 TI - Live HeLa cells preconcentrate and differentiate inorganic arsenic species. AB - Live HeLa cells immobilized on Sephadex G-50 beads were used as a medium for the preconcentration and speciation of inorganic arsenic. The sorption of arsenic species by live HeLa cells involves both surface uptake and bioaccumulation within the cells. At pH 3.0, the cells accumulate arsenate with high specificity over arsenite: 83.0 +/- 1.3% of the arsenate was sorbed while the retention of arsenite was negligible at 2.1 +/- 0.6%. The speciation of inorganic arsenic could thus be performed by direct determination of arsenate followed by quantifying total inorganic arsenic after conversion of arsenite to arsenate. We formed a disposable live cell preconcentration microcolumn with the live HeLa cells immobilized on Sephadex G-50 beads. After the sample was passed through the column for sorption to occur, the cells and any retained arsenate were stripped with 2 M HNO(3). The arsenic in the 30 microL eluate was directly measured by graphite furnace atomic absorption spectrometry. A new microcolumn was used for each sample. With a sample volume of 450 muL, a S/N = 3 limit of detection (LOD) of 0.05 microg/L and a linear range of 0.15-2.5 microg/L were attained; the relative standard deviation (RSD) was 1.7% at 1.25 microg/L. The procedure was validated by arsenic speciation in certified reference river water. PMID- 19128048 TI - Ytterbium(III) triflate-catalyzed amination of 1-cyclopropylprop-2-yn-1-ols as an expedient route to conjugated enynes. AB - Ytterbium(III) triflate-catalyzed ring opening of substituted 1-cyclopropyl-2 propyn-1-ols with sulfonamides as an efficient synthetic route to conjugated enynes is described herein. The reaction was operationally straightforward and accomplished in moderate to good yields and regioselective manner in all except one case under mild conditions. PMID- 19128049 TI - Glycomic characterization of prostate-specific antigen and prostatic acid phosphatase in prostate cancer and benign disease seminal plasma fluids. AB - Prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) are glycoproteins secreted by prostate epithelial cells, and have a long clinical history of use as serum biomarkers of prostate cancers. These two proteins are present at significantly higher concentrations in seminal plasma, making this proximal fluid of the prostate a good source for purifying enough protein for characterization of prostate disease associated changes in glycan structures. With the use of seminal fluid samples representative of normal control, benign prostatic disease and prostate cancers, PAP and PSA were enriched by thiophilic absorption chromatography. Released N-linked glycan constituents from both proteins were analyzed by a combination of normal phase HPLC and MALDI-TOF spectrometry. For PSA, 40 putative glycoforms were determined, and 21 glycoforms were determined for PAP. PAP glycans were further analyzed with a hybrid triple quadrupole/linear ion trap mass spectrometer to assign specific glycoform classes to each of the three N-linked sites. The glycans identified in these studies will allow for more defined targeting of prostate disease-specific changes for PAP, PSA and other secreted prostatic glycoproteins. PMID- 19128050 TI - Transzygomatic extended middle fossa approach for upper petroclival skull base lesions. AB - Central skull base lesions in the upper retroclival and petroclival regions can be challenging to access because of their location anterior to the brainstem. Several transpetrosal approaches have been developed to access the petroclival junction, including anterior petrosal (anterior petrosectomy), posterior petrosal (retrolabyrinthine, translabyrinthine, transcochlear), and combined petrosal approaches. The anterior petrosal approach is best suited for upper petroclival lesions located anterior and superior to the internal auditory canal and superior to the inferior petrosal sinus. This approach provides direct access to the anteromedial cerebellopontine angle, petrous apex, Meckel cave, and ventrolateral brainstem between the trigeminal root and the facial nerve. The authors describe their modification of an anterior petrosal approach, the so-called transzygomatic extended middle fossa approach, which incorporates a zygomatic osteotomy, anterior mobilization of the V3, and extensive middle fossa drilling. This exposure provides a wider surgical corridor for direct view of the clivus and ventral brainstem. PMID- 19128051 TI - Electrochemical and spectral properties of ferrocene (Fc) in ionic liquid: 1 butyl-3-methylimidazolium triflimide, [BMIM][NTf(2)]. Concentration effects. AB - Several earlier studies of the electrochemical oxidation of ferrocene (Fc) in room-temperature ionic liquids revealed an essentially nonlinear dependence of the oxidation current on the Fc concentration in its relatively dilute solutions, with its formally calculated diffusion coefficient strongly increasing with the concentration. Since no plausible mechanism leading to this very unusual finding had been proposed, our study of Fc solutions in 1-butyl-3-methylimidazolium triflimide, [BMIM][NTf(2)], was performed to verify whether the above observation originated from an incorrect determination of the dissolved Fc concentration. Our observations have demonstrated that reliable control of the Fc concentration in solution is complicated by factors such as the low amount of Fc used to prepare small-volume solutions or the great difficulty to dissolve completely a solid powder in a solvent with an extremely high viscosity. An unexpected additional complication is related to a sufficiently high volatility of Fc which manifests itself even at room temperature and especially at elevated temperatures or/and in the course of vacuum treatment of its solutions or its solid powder. Parallel measurements of electrochemical responses and UV-visible spectra for several series of Fc solutions of various concentrations (prepared with the use of different procedures) have shown a perfect parallelism between the peak current and the intensity of the absorption band in the range of 360-550 nm, leading us to the conclusion of a linear relationship between the oxidation current and the molecularly dissolved Fc concentration. The relations of these measured characteristics with the estimated Fc concentration in these solutions have demonstrated a much greater dispersion (attributed to the difficulty of a precise measurement of the latter) but without a significant deviation from the linearity in general. This finding has allowed us to estimate the diffusion coefficient of this species: D = (1.7 +/- 0.2) x 10(-7) cm(2)/s. The extinction coefficients for the maximum of the absorption band (at 440 nm) of Fc have been compared for a series of solvents: [BMIM][NTf(2)], acetonitrile, THF, heptane, CH(2)Cl(2), ethanol, and toluene. A simple method to estimate reliably the concentration of solute Fc in ionic liquids based on spectroscopic measurements has been proposed, owing to the proximity of Fc absorption properties for a great variety of solvents. PMID- 19128052 TI - Alkyl chain interaction at the surface of room temperature ionic liquids: systematic variation of alkyl chain length (R = C(1)-C(4), C(8)) in both cation and anion of [RMIM][R-OSO(3)] by sum frequency generation and surface tension. AB - The gas-liquid interface of halide-free 1,3-dialkylimidazolium alkyl sulfates [RMIM][R-OSO(3)] with R chain length from C(1)-C(4) and C(8) has been studied systematically using the surface-specific sum frequency generation (SFG) vibrational spectroscopy and surface tension measurements. From the SFG spectra, vibrational modes from the methyl group of both cation and anion are observed for all ionic liquid samples considered in the present study. These results suggest the presence of both ions at the gas-liquid interface, which is further supported by surface tension measurements. Surface tension data show a decreasing trend as the alkyl chain in the imidazolium cation is varied from methyl to butyl chain, with a specific anion. A similar trend is observed when the alkyl chain of the anion is modified and the cation is fixed. PMID- 19128054 TI - The effect of the primary solvate shell on the mechanism of the Stober silica synthesis. A density functional investigation. AB - The target of the present computational study was the acid catalyzed bond cleavage of the Si-O and C-O bonds in siloxane, alkoxysilane and ether in aqueous media. In the present study the effect of water as a solvent has been modeled using a full primary solvate shell built up from water molecules connected via hydrogen bonds around the reacting molecules. The interaction energy between the embedding water cluster and the "solvated" molecule gives an estimate for solution effects. The cleavage of the Si-O bonds in these molecular clusters proceeds with low barriers; furthermore the reaction energies corrected with the solvent interaction energies gives a reaction thermodynamics, which is in accordance with the experimental results. Molecules with a Si-O bond form stable pentavalent silicon with the solvent water molecules if protonated, while in the case of the neutral molecules tetracoordinate silicon is obtainable. The summary of the calculated reaction paths gives a possible route of siloxane formation from methoxysilane in aqueous media. The same computational methodology predicts that the hydrolysis of dimethyl ether is hindered by a substantial barrier. PMID- 19128053 TI - Unusually reactive and selective carbonyl ylides for three-component cycloaddition reactions. AB - Conditions are described for the Rh-catalyzed formation of highly functionalized dihydro- and tetrahydrofuran products via three-component reactions of aldehydes, alpha-alkyl-alpha-diazoesters, and dipolarophiles. The alkyl-substituted carbonyl ylides that are generated in this fashion are highly reactive in cycloaddition reactions and display a scope of reactivity that is much broader than the three component reactions of carbonyl ylides derived from ethyl diazoacetate or alpha aryl-alpha-diazoesters. The reactions of alkyl-substituted carbonyl ylides proceed with high regioselectivity and diastereoselectivity that are rationalized in terms of an asynchronous, endo-selective transition state. PMID- 19128055 TI - Pyrazolo[1,5-a]-1,3,5-triazine as a purine bioisostere: access to potent cyclin dependent kinase inhibitor (R)-roscovitine analogue. AB - Pharmacological inhibitors of cyclin-dependent kinases (CDKs) have a wide therapeutic potential. Among the CDK inhibitors currently under clinical trials, the 2,6,9-trisubstituted purine (R)-roscovitine displays rather high selectivity, low toxicity, and promising antitumor activity. In an effort to improve this structure, we synthesized several bioisosteres of roscovitine. Surprisingly, one of them, pyrazolo[1,5-a]-1,3,5-triazine 7a (N-&-N1, GP0210), displayed significantly higher potency, compared to (R)-roscovitine and imidazo[2,1-f] 1,2,4-triazine 13 (N-&-N2, GP0212), at inhibiting various CDKs and at inducing cell death in a wide variety of human tumor cell lines. This approach may thus provide second generation analogues with enhanced biomedical potential. PMID- 19128056 TI - Synthesis and evaluation of partly fluorinated block copolymers as MRI imaging agents. AB - A series of well-defined diblock copolymers of acrylic acid with partially fluorinated acrylate and methacrylate monomers were synthesized using ATRP as potential 19F MRI imaging agents. The diblock copolymers could undergo spontaneous self-assembly in mixed and aqueous solvents to form stable micelles with a diameter from approximately 20-45 nm, having a fluorine-rich core that provides a strong signal for MRI examinations. The observed MRI image intensities were related to the NMR longitudinal and transverse relaxation times, and were found to depend on polymer structure and method of micellization. Two distinct T2 relaxation times were measured; on comparison of expected MRI image intensities with those observed experimentally, it was found that methacrylate polymers show systematically lower signal intensity than acrylate polymers. This is related to the presence of a population of nuclear spins having very short T2 relaxation times that cannot be detected under high-resolution NMR and MRI conditions. PMID- 19128057 TI - Self-assembly of genetically engineered spider silk block copolymers. AB - The design, construction, and preliminary characterization of a novel family of spider silk-like block copolymers are described. The design was based on the assembly of individual spider silk modules, in particular, polyalanine (A) and glycine-rich (B) blocks, that display different phase behavior in aqueous solution. Spider silk was chosen as a model for these block copolymer studies based on its extraordinary material properties, such as toughness, biocompatibility, and biodegradability. Trends in spider silk-like block copolymer secondary structure and assembly behavior into specific material morphologies were determined as a function of the number of hydrophobic blocks, the presence of a hydrophilic purification tag and solvent effects. Structures and morphologies were assessed by Fourier transform infrared spectroscopy and scanning electron microscopy. In terms of structure, beta-sheet content increased with an increase in the number of polyalanine blocks, and the purification tag had significant impact on the secondary structure. In terms of morphology, spheres, rod-like structures, bowl-shaped micelles, and giant compound micelles were observed and the morphologies were linked with the size of the hydrophobic block, the presence of the purification tag, and the solvent environment. This study provides a basis for future designs of smart biomaterials based on spider silk chemistries, with controlled structure-architecture-function relationships. PMID- 19128058 TI - Compared reactivity of allyl ribosides in UV-initiated free radical copolymerization with acceptor monomers. AB - The free radical copolymerization of allyl ribosides with diethyl fumarate and maleate was investigated for evaluating the potential of donor-acceptor type copolymerization applied to unsaturated monomers derived from renewable feedstock. The photochemically induced polymerization of model monomer blends was conducted in solution as well as in liquid films of bulk reactants. Infrared spectroscopy was used to monitor the consumption of the allylic donor monomer and of the butenedicarboxylate acceptor monomers. The method allowed examining the influence of the nature of the monomer pair and of their relative concentration on the kinetic profiles. Comparison with reference vinyl ether monomers confirmed the expected lower reactivity of the blends containing allylic derivatives. SEC and NMR analysis supported the occurrence of degradative chain transfer during the reactions involving allylic monomers. However, allyl derivatives of glycerol as well as O-allyl ribosides were shown to undergo polymerization with high conversion of both monomers when blended in appropriate molar ratios. PMID- 19128059 TI - Femtosecond laser-induced ionization/dissociation of protonated peptides. AB - Although tandem mass spectrometry has revolutionized the identification and structural characterization of peptides and proteins, future advances in comprehensive proteome analysis will depend on the development of improved methods for ion activation that yield greater sequence information, and with selective control over the fragmentation chemistry. This report presents initial findings that demonstrate the utility of a novel ion activation method using ultrashort (approximately 30 fs) laser pulses as a means to overcome the limitations of current technologies, while opening the door to solving significant challenges in protein and peptide analysis. PMID- 19128060 TI - Redox-activating dip-pen nanolithography (RA-DPN). AB - Dip pen nanolithography (DPN) involves the direct transfer of an ink from a coated atomic force microscope (AFM) tip to a substrate of interest and uses as many as 55,000 pens to form arbitrary patterns of alkanethiols, oligonucleotides, proteins, and viruses. Two limitations of DPN are the difficulty in transporting high molecular weight inks and the need to optimize individually the transport rates and tip inking methods of each molecule. As an alternative strategy that circumvents these two challenges, a method termed redox activating DPN (RA-DPN) is reported. In this strategy, an electrochemically active, quinone functionalized surface is toggled from the reduced hydroquinone form to the oxidized benzoquinone form by the delivery of an oxidant by DPN. While the benzoquinone form is susceptible to nucleophilic attack in Michael-type additions, hydroquinone is not and acts as a passivating agent. Because both forms of the quinone are kinetically stable, the patterned surface can be immersed in a solution of a target containing any strong nucleophile, which will react only where the benzoquinone form persists on the surface. For proof-of concept demonstrations, quinone surfaces were patterned by the delivery of the oxidant cerric ammonium nitrate and were immersed in solutions of AF549 labeled cholera toxin beta subunit or oligonucleotides modified at the 5' end with an amine and the 3' end with a fluorophore. Fluorescent patterns of both the proteins and oligonucleotides were observed by epifluorescence microscopy. Additionally, RA-DPN maintains the advantageous ability of DPN to control feature size by varying the dwell time of the tip on the surface, and variation of this parameter has resulted in feature sizes as small as 165 nm. With this resolution, patterns of 50,000 spots could be made in a 100 x 100 microm(2) grid. PMID- 19128061 TI - Mechanistic insights into the rhodium-catalyzed intramolecular ketone hydroacylation. AB - [Rh((R)-DTBM-SEGPHOS)]BF(4) catalyzes the intramolecular hydroacylation of ketones to afford seven-membered lactones in large enantiomeric excess. Herein, we present a combined experimental and theoretical study to elucidate the mechanism and origin of selectivity in this C-H bond activation process. Evidence is presented for a mechanistic pathway involving three key steps: (1) rhodium(I) oxidative addition into the aldehyde C-H bond, (2) insertion of the ketone CO double bond into the rhodium hydride, and (3) C-O bond-forming reductive elimination. Kinetic isotope effects and Hammett plot studies support that ketone insertion is the turnover-limiting step. Detailed kinetic experiments were performed using both 1,3-bis(diphenylphosphino)propane (dppp) and (R)-DTBM SEGPHOS as ligands. With dppp, the keto-aldehyde substrate assists in dissociating a dimeric precatalyst 8 and binds an active monomeric catalyst 9. With [Rh((R)-DTBM-SEGPHOS)]BF(4), there is no induction period and both substrate and product inhibition are observed. In addition, competitive decarbonylation produces a catalytically inactive rhodium carbonyl species that accumulates over the course of the reaction. Both mechanisms were modeled with a kinetics simulation program, and the models were consistent with the experimental data. Density functional theory calculations were performed to understand more elusive details of this transformation. These simulations support that the ketone insertion step has the highest energy transition state and reveal an unexpected interaction between the carbonyl-oxygen lone pair and a Rh d-orbital in this transition state structure. Finally, a model based on the calculated transition state geometry is proposed to rationalize the absolute sense of enantioinduction observed using (R)-DTBM-SEGPHOS as the chiral ligand. PMID- 19128068 TI - Vaccines and autism: a tale of shifting hypotheses. AB - Although child vaccination rates remain high, some parental concern persists that vaccines might cause autism. Three specific hypotheses have been proposed: (1) the combination measles-mumps-rubella vaccine causes autism by damaging the intestinal lining, which allows the entrance of encephalopathic proteins; (2) thimerosal, an ethylmercury-containing preservative in some vaccines, is toxic to the central nervous system; and (3) the simultaneous administration of multiple vaccines overwhelms or weakens the immune system. We will discuss the genesis of each of these theories and review the relevant epidemiological evidence. PMID- 19128090 TI - On-flow pulsed field gradient heteronuclear correlation spectrometry in off-line LC-SPE-NMR analysis of chemicals related to the chemical weapons convention. AB - Hyphenation of liquid chromatography with nuclear magnetic resonance spectroscopy (LC-NMR) is a useful technique in the analysis of complex samples. However, application of on-flow 1H NMR spectrometry during the LC-NMR analysis usually suffers from high intensity of eluent resonances. The poor dynamic range can be improved either with use of deuterated eluents or with various signal suppression schemes. Deuterated eluents are expensive, and peak-selective signal suppression schemes are often unsatisfactory when detection of chemicals at low concentration is needed. If the analytes have a common heteronucleus, on-flow pulsed field gradient heteronuclear correlation spectrometry can offer several benefits. The analytes can be monitored selectively, while the intense nondeuterated eluent and impurity background can be effectively eliminated. In our study, on-flow one dimensional (1D) 1H-31P heteronuclear single quantum coherence (HSQC) spectrometry was utilized in the analysis of characteristic organophosphorus degradation products of nerve agents sarin and soman during chromatographic separation. These chemicals were not detectable by UV, so their retention times were monitored using on-flow 1D 1H-31P HSQC. This enabled application of LC-NMR combined with solid-phase extraction (LC-SPE-NMR) in analysis of these organophosphorus chemicals in an alkaline decontamination solution. The analytes were extracted from the SPE cartridges with deuterated eluent, and the off-line NMR analysis was performed using a mass-sensitive microcoil probe head. The used on-flow 1D 1H-31P HSQC approach offered a high dynamic range and good detection limit (ca. 10 microg/55 nmol) with a high sampling frequency (1 point per 2 s) in the acquired pseudo-two-dimensional spectrum. No significant impurity background was present in the off-line NMR samples, and identification of the extracted analytes was straightforward. PMID- 19128143 TI - Feasibility of a clinical chemical analysis approach to predict misuse of growth promoting hormones in cattle. AB - A study was performed to determine if targeted metabolic profiling of cattle sera could be used to establish a predictive tool for identifying hormone misuse in cattle. Metabolites were assayed in heifers (n = 5) treated with nortestosterone decanoate (0.85 mg/kg body weight), untreated heifers (n = 5), steers (n = 5) treated with oestradiol benzoate (0.15 mg/kg body weight) and untreated steers (n = 5). Treatments were administered on days 0, 14, and 28 throughout a 42 day study period. Two support vector machines (SVMs) were trained, respectively, from heifer and steer data to identify hormone-treated animals. Performance of both SVM classifiers were evaluated by sensitivity and specificity of treatment prediction. The SVM trained on steer data achieved 97.33% sensitivity and 93.85% specificity while the one on heifer data achieved 94.67% sensitivity and 87.69% specificity. Solutions of SVM classifiers were further exploited to determine those days when classification accuracy of the SVM was most reliable. For heifers and steers, days 17-35 were determined to be the most selective. In summary, bioinformatics applied to targeted metabolic profiles generated from standard clinical chemistry analyses, has yielded an accurate, inexpensive, high throughput test for predicting steroid abuse in cattle. PMID- 19128144 TI - Aptamer-modified micellar electrokinetic chromatography for the enantioseparation of nucleotides. AB - In this paper, a new aptamer-based capillary electrophoresis (CE) method, which was able to separate the enantiomers of an anionic target (adenosine monophosphate, AMP) displaying the same electrophoretic mobility as that of the oligonucleotidic chiral selector, is reported. The design of the aptamer-modified micellar electrokinetic chromatography (MEKC) mode consisted of nonionic micelles which acted as a pseudostationary phase and a hydrophobic cholesteryl group tagged aptamer (Chol-Apt) which partitioned into the uncharged micellar phase. Under partial-filling format and suppressed electroosmotic flow conditions, the strong mobility alteration of Chol-Apt permitted AMP enantiomers to pass through the micelle-anchored aptamer zone and promoted the target enantioseparation. The influence of several electrophoretic parameters (such as concentration and nature of the nonionic surfactant, preincubation of the Chol-Apt and surfactant, capillary temperature, and applied voltage) on the AMP enantiomer migration was investigated in order to define the utilization conditions of the aptamer modified MEKC mode. The chiral resolution, in a single run, of three adenine nucleotides, i.e., AMP, ADP (adenosine diphosphate), and ATP (adenosine triphosphate), was further accomplished using such methodology. This approach demonstrates the possibility to extend the CE applicability of aptamer chiral selectors to potentially any target, without restriction on its charge-to-mass ratio. PMID- 19128146 TI - "Nanoparticle-in-alloy" approach to efficient thermoelectrics: silicides in SiGe. AB - We present a "nanoparticle-in-alloy" material approach with silicide and germanide fillers leading to a potential 5-fold increase in the thermoelectric figure of merit of SiGe alloys at room temperature and 2.5 times increase at 900 K. Strong reductions in computed thermal conductivity are obtained for 17 different types of silicide nanoparticles. We predict the existence of an optimal nanoparticle size that minimizes the nanocomposite's thermal conductivity. This thermal conductivity reduction is much stronger and strikingly less sensitive to nanoparticle size for an alloy matrix than for a single crystal one. At the same time, nanoparticles do not negatively affect the electronic conduction properties of the alloy. The proposed material can be monolithically integrated into Si technology, enabling an unprecedented potential for micro refrigeration on a chip. High figure-of-merit at high temperatures (ZT approximately 1.7 at 900 K) opens up new opportunities for thermoelectric power generation and waste heat recovery at large scale. PMID- 19128147 TI - Organic polymer supports for synthesis and for reagent and catalyst immobilization. PMID- 19128145 TI - A single-nucleotide polymorphism in the gene encoding osteoprotegerin, an anti inflammatory protein produced in response to infection with diarrheagenic Escherichia coli, is associated with an increased risk of nonsecretory bacterial diarrhea in North American travelers to Mexico. AB - BACKGROUND: Osteoprotegerin (OPG), an immunoregulatory member of the TNF receptor superfamily, is expressed in inflamed intestinal mucosa. We investigated whether OPG is produced by intestinal epithelial cells and tested the hypothesis that single-nucleotide polymorphisms (SNPs) in the gene encoding OPG (TNFRSF11B) are associated with traveler's diarrhea (TD) among North American travelers to Mexico. METHODS: OPG concentration was measured in the supernatants of T84 cells infected with various diarrheagenic Escherichia coli pathotypes. Genotyping was performed for 4 SNPs in the OPG gene for 968 North American travelers with or without TD. Stool samples from travelers with TD were evaluated for the presence of enteric pathogens. RESULTS: T84 cells produced higher OPG levels in response to infection with various diarrheagenic E. coli pathotypes than with E. coli controls (P<.05). A SNP in the exon 1 region of the OPG gene (OPG+1181G>C) was associated with TD in white travelers who stayed in Mexico for >1 week during the summer (P=.009) and for TD due to nonsecretory pathogens (P=.001). CONCLUSIONS: Our study suggests that OPG is secreted by intestinal epithelial cells in response to enteropathogens and that a polymorphism in the OPG gene is associated with an increased susceptibility to TD. PMID- 19128148 TI - Biotransformation of two ent-Pimara-9(11),15-diene derivatives by Gibberella fujikuroi. AB - The incubation of 19-hydroxy-13-epi-ent-pimara-9(11),15-diene (4) with Gibberella fujikuroi gave 8 alpha,19-dihydroxy-9 alpha,11alpha-epoxy-13-epi-ent-pimara-15 ene (6), 7-oxo-11 alpha,19-dihydroxy-13-epi-ent-pimara-8(9),15-diene (7), 7-oxo 11beta,19-dihydroxy-13-epi-ent-pimara-8(9),15-diene (9), and 8 alpha,19-dihydroxy 9 alpha,11 alpha:15,16-diepoxy-13-epi-ent-pimarane (11), while the feeding of 13 epi-ent-pimara-9(11),15-diene-19-oic acid (5) with this fungus afforded 1-oxo-2 alpha,9 alpha-dihydroxy-13-epi-ent-pimara-11,15-dien-19-oic acid (13), 1-oxo-2 beta,9 alpha-dihydroxy-13-epi-ent-pimara-11,15-dien-19-oic acid (14), 13-epi-ent pimara-9(11),15-dien-1,19-dioic acid 1,2-lactone (15), and 1-oxo-12 beta-hydroxy 13-epi-ent-pimara-9(11),15-dien-19-oic acid (16). In both biotransformations, the main reaction was the epoxidation of the 9(11)-double bond, followed by rearrangement to afford allylic alcohols. The formation of lactone 15 represents the first time that a Baeyer-Villiger oxidation has been observed in a microbiological transformation with this fungus. PMID- 19128149 TI - Mechanistic aspects of alkyne migration in alkylidene carbenoid rearrangements. AB - The mechanism of the Fritsch-Buttenberg-Wiechell rearrangement of (13)C labeled precursors has been examined to determine the propensity of the alkynyl (R-CC-) group to migrate in an alkylidene carbenoid species. Reaction of dibromoolefins with n-BuLi and ketones with Me(3)SiC(Li)N(2) both demonstrate that the alkynyl moiety readily undergoes 1,2-migration from carbenoid intermediates. PMID- 19128150 TI - Design, synthesis, and application of enantioselective coupling reagent with a traceless chiral auxiliary. AB - Stable chiral N-triazinylbrucinium tetrafluoroborate enantioselectively activates racemic carboxylic acids yielding enantiomerically enriched amides, esters, and dipeptides with er from 8:92 to 0.5:99.5. Due to the departure of a chiral auxiliary after the activation of the carboxylic function, all of the subsequent stages of the coupling reaction proceed without any perturbation caused by a chirality discriminator (traceless). Therefore, the advantageous coupling conditions, configuration, and enantiomeric purity of the final product are entirely predictable from the model experiment. PMID- 19128151 TI - Catalyst-free growth of quasi-aligned nanorods of single crystal Cu3Mo2O9 and their catalytic properties. AB - We propose a simple and catalyst-free method to grow quasi-aligned single crystalline Cu(3)Mo(2)O(9) nanorods in terms of a mechanism differing from the conventional vapor-solid (VS) and vapor-liquid-solid (VLS) ones for chemical vapor deposition (CVD) methods by directly heating Cu foils in a mixed atmosphere of MoO(3) vapor and air. High quality Cu(3)Mo(2)O(9) nanorods can be simply grown in a temperature range from 450 to 550 degrees C whose diameter, length, and distribution density are dependent on both heating temperature and time. Interestingly, the growth rate at 550 degrees C drops significantly after 6 h. All nanorods grow along the [010] direction. On the basis of a proposed growth model, the nucleation of Cu(3)Mo(2)O(9) nanorods is believed to be governed by formation of initial polycrystalline Cu(x)O protuberances with nanoscale diameters on Cu foils which may act as growth "templates". This novel method can be applied to grow other similar tertiary transition metal oxide nanostructures on substrates with large sizes. Most importantly, these Cu(3)Mo(2)O(9) nanorods decrease the ignition temperature of Printex U model soot from 600 to 438 degrees C, being in between 200 and 450 degrees C of the exhaust of diesel-powered combustion engines, which are therefore expected to be a potential efficient and environmentally friendly catalyst for diesel exhaust combustion. PMID- 19128152 TI - A bioaccumulative cyclometalated platinum(II) complex with two-photon-induced emission for live cell imaging. AB - The cyclometalated platinum(II) complex [Pt(L)Cl], where HL is a new cyclometalating ligand 2-phenyl-6-(1H-pyrazol-3-yl)pyridine containing C(phenyl), N(pyridyl), and N(pyrazolyl) donor moieties, was found to possess two-photon induced luminescent properties. The two-photon-absorption cross section of the complex in N,N-dimethylformamide at room temperature was measured to be 20.8 GM. Upon two-photon excitation at 730 nm from a Ti:sapphire laser, bright-green emission was observed. Besides its two-photon-induced luminescent properties, [Pt(L)Cl] was able to be rapidly accumulated in live HeLa and NIH3T3 cells. The two-photon-induced luminescence of the complex was retained after live cell internalization and can be observed by two-photon confocal microscopy. Its bioaccumulation properties enabled time-lapse imaging of the internalization process of the dye into living cells. Cytotoxicity of [Pt(L)Cl] to both tested cell lines was low, according to MTT assays, even at loadings as high as 20 times the dose concentration for imaging for 6 h. PMID- 19128154 TI - Co(II) molecular square with single-molecule magnet properties. AB - A new tetranuclear cobalt(II) molecular square in which adjacent Co(II) centers are linked by a mu(2)-bridging oxygen atom and a N-N bridge along the edges of the square has been designed for single-molecule magnets (SMMs) with high anisotropy barriers. The overall intramolecular ferromagnetic coupling at low temperature combined with the slow relaxation at static zero fields suggests a SMM behavior for this molecular square. The zero-field cooled magnetization (ZFCM) and field cooling magnetization (FCM) at 10 Oe illustrate the nonreversibility and bifurcation below 4.5 K. The deviations of magnetization from the saturated value in strong applied fields demonstrate the participation of low-lying excited states. The peaks of the out-of-phase signals are observed corresponding to coincidence of the applied ac field oscillation frequency with the relaxation rate. PMID- 19128153 TI - Structural models for the active site of acetyl-CoA synthase: synthesis of dinuclear nickel complexes having thiolate, isocyanide, and thiourea on the Ni(p) site. AB - The trinuclear nickel complex [{Ni(dadt(Et))}(2)Ni](NiBr(4)) (dadt(Et) = N,N' diethyl-3,7-diazanonane-1,9-dithiolate) (1a), prepared by the reaction of Ni(dadt(Et)) and Ni(EtOH)(4)Br(2), was found to serve as a useful synthetic precursor of various dinuclear nickel complexes modeling the active site of acetyl-CoA synthase (ACS). The reactions of 1a with 4 equiv of the potassium salts of arenethiolates in ethanol produced a series of dinuclear nickel thiolate complexes Ni(dadt(Et))Ni(SAr)(2) (Ar = Ph (2a), p-Tol (2b), 2,4,6 triisopropylphenyl (Tip) (2c)) in good yields. The analogous reactions of 1a with Ag(OTf) in the presence of (t)BuNC and (NMe(2))(2)CS (tmtu) generated the dicationic dinuclear nickel complexes [Ni(dadt(Et))Ni((t)BuNC)(2)](OTf)(2) (3) and [Ni(dadt(Et))Ni(tmtu)(2)](OTf)(2) (4), respectively. The molecular structures of 1a, 2a-c, 3, and 4 determined by X-ray analysis compare well with that of A cluster in ACS. PMID- 19128155 TI - Synthesis, structure, and magnetism of heterobimetallic trinuclear complexes {[L2Co2Ln][X]} [Ln = Eu, X = Cl; Ln = Tb, Dy, Ho, X = NO3; LH3 = (S)P[N(Me)N=CH C6H3-2-OH-3-OMe]3]: A 3d-4f family of single-molecule magnets. AB - Sequential reaction of LH3 (LH3 = (S)P[N(Me)N=CH-C6H3-2-OH-3-OMe]3) with Co(OAc)2 x 4 H2O followed by reaction with lanthanide salts afforded trinuclear heterobimetalllic compounds {[L2Co2Ln][X]} [Ln = Eu (1), X = Cl; Ln = Tb (2), Dy (3), Ho (4), X = NO3] in excellent yields. These compounds retain their integrity in solution as determined by electrospray ionization mass spectrometry studies. The molecular structures of 1-4 were confirmed by a single-crystal X-ray structural study and reveal that these are isostructural. In all of the compounds, the three metal ions are arranged in a perfectly linear manner and are held together by two trianionic ligands, L3-. The two terminal Co(II) ions contain a facial coordination environment (3N, 3O) comprising three imino nitrogen atoms and three phenolate oxygen atoms. The coordination geometry about the cobalt atom is severely distorted. An all-oxygen coordination environment (12O) is present around the central lanthanide ion, which is present in a distorted icosahedral geometry. The coordination sphere around the lanthanide ion is achieved by utilizing three phenolate oxygen atoms and three methoxy oxygen atoms of each ligand. In all of these trinuclear complexes (1-4), the Co-Ln distances are around 3.3 A, while the Co-Co distances range from 6.54 to 6.60 A. The screw-type coordination mode imposed by the ligand induces chirality in the molecular structure, although all of the complexes crystallize as racemates. Magnetic properties of 1-4 have been studied in detail using dc and ac susceptibility measurements. Dynamic measurements reveal that 2-4 display a single-molecule magnet behavior, while the Co2Eu (1) analogue does not show any out-of-phase ac susceptibility. PMID- 19128156 TI - Cytotoxic spirostane-type saponins from the roots of Chlorophytum borivilianum. AB - Four new spirostane-type saponins named borivilianosides E-H (1-4) were isolated from an ethanol extract of the roots of Chlorophytum borivilianum together with two known steroid saponins (5 and 6). The structures of 1-4 were elucidated using mainly 2D NMR spectroscopic techniques and mass spectrometry. The cytotoxicity of borivilianosides F (2), G (3), and H (4) and three known compounds was evaluated using two human colon cancer cell lines (HT-29 and HCT 116). PMID- 19128157 TI - Ionic liquids containing boron cluster anions. AB - The combination of different boron cluster anions and some of the cations typically found in the composition of ionic liquids has been possible by straightforward metathetic reactions, producing new low melting point salts; the imidazolium cations have been systematically studied, [C(n)mim]+ (when [C(n)mim]+ = 1-alkyl-3-methylimidazolium; n = 2, 4, 6, 8, 10, 12, 14, 16, or 18). Melting points increase in the anionic order [Co(C2B9H11)2]- < [C2B9H12]- < [B10Cl10]2- < [B12Cl12]2-. Nevertheless, alkyl chain length dramatically influences the thermal behavior, suggesting that packing inefficiency is the main cause of the existence of room temperature ionic liquids. The salts [C(n)mim][Co(C2B9H11)2] (n = 4, 6, 8, 10, 12 or 14) are liquids at room temperature, presenting strikingly low glass transition temperatures (> or = -34 degrees C). The salts [C(n)mim]2[X] ([X]2- = [B10Cl10]2- or [B12Cl12]2-, n = 16 or 18) show liquid crystal phases between the solid and liquid states. Tetraalkylphosphonium salts of [B10Cl10]2- have also been prepared. Physical properties, such as thermal stability, density, or viscosity, have been measured for some selected samples. The presence of the perhalogenated dianion [B12Cl12]2- in the composition of the imidazolium salts renders highly thermally stable compounds. For example, [C2mim]2[B12Cl12] starts to decompose above 480 degrees C in a dynamic TGA analysis under a dinitrogen atmosphere. Crystal structures of [C2mim][Co(C2B9H11)2] and [C2mim]2[B12Cl12] have been determined. 1H NMR spectra of selected imidazolium-boron cluster anion salts have been recorded from solutions as a function of the concentration, showing trends related to the cation-anion interactions. PMID- 19128159 TI - Upper gastrointestinal bleeding in a patient with HIV infection. PMID- 19128162 TI - Class of antiretroviral therapy and CD4+ T cell count recovery: independence questioned. PMID- 19128163 TI - Fatal acute varicella-zoster virus hemorrhagic meningomyelitis with necrotizing vasculitis in an HIV-infected patient. PMID- 19128164 TI - Molecular evidence of patient-to-patient transmission of hepatitis E virus in a hematology ward. PMID- 19128167 TI - Phenanthroline decorated by a crown ether as a module for metallorganic polyoxometalate hybrid catalysts: the Wacker type oxidation of alkenes with nitrous oxide as terminal oxidant. AB - A 1,10-phenanthroline ligand decorated at the 5,6-position by a 15-crown-5 ether moiety was prepared. Ligation of Pd(II) at the nitrogen atom positions followed by complexation at the crown ether group of a redox active H(5)PV(2)Mo(10)O(40) polyoxometalate yielded a hybrid metallorganic-polyoxometalate complex, Pd(II)(15 crown-5-phen)Cl(2)-H(5)PV(2)Mo(10)O(40). This complex was characterized by IR, UV vis, ESI-MS, and NMR spectroscopy and elemental analysis that all confirmed the hybrid nature of the complex. Pd(II)(15-crown-5-phen)Cl(2)-H(5)PV(2)Mo(10)O(40) was used as a catalyst for the Wacker type oxidation of 1-alkenes to yield the corresponding methylketones in essentially quantitative yields using nitrous oxide as the terminal oxidant. PMID- 19128168 TI - N-N bond cleavage in diazoalkanes by a bis(imino)pyridine iron complex. AB - Addition of monosubstituted diazoalkanes, N(2)CHR (R = Ph, p-Tolyl, (t)Bu, Cy), to the bis(imino)pyridine iron dinitrogen complex, ((iPr)PDI)Fe(N(2))(2), at 23 degrees C in benzene-d(6) resulted in rapid cleavage of the N-N bond to yield the iron nitrile and imine complexes, ((iPr)PDI)FeNCR and ((iPr)PDI)FeHNCHR, respectively. Aryl azines were also cleaved with ((iPr)PDI)Fe(N(2))(2) and yielded the same products. Mechanistic studies support a pathway involving initial formation of an iron alkylidene followed by rapid [4pi + 2pi] cycloaddition with another equivalent of diazoalkane. Subsequent rate-determining 1,3-hydrogen migration followed by retrocycloaddition yields the observed products. PMID- 19128169 TI - N-CN bond cleavage of cyanamides by a transition-metal complex. AB - N-CN bond cleavage of cyanamides (R(2)N-CN) has been attained at room temperature in the reaction of R(2)N-CN with Cp(CO)(2)Fe(SiEt(3)). The mechanistic investigation revealed that silyl migration from Fe to CN nitrogen of cyanamide gave an N-silylated eta(2)-amidino iron complex, which was isolated and characterized by X-ray analysis. Catalytic N-CN bond cleavage was also attained using a methyl molybdenum complex under thermal conditions. PMID- 19128170 TI - Hydrogen-bonded ferroelectrics based on metal-organic coordination. AB - Metal-organic coordination (MOC)-type ferroelectrics, cobalt(II) (R)-2 methylpiperazine (MPPA) trichloride [Co(II)Cl(3)(H-MPPA)], was constructed through hydrogen bonds. It is a good ferroelectric candidate with a P(s) = 6.8 microC.cm(-2) as high as almost twice that of triglycine sulfate (P(s) = 3.5 microC.cm(-2)) and significantly larger than that of KH(2)PO(4) at the low temperature ferroelectric phase Fdd2. [Co(II)Cl(3)(H-MPPA)] is the first example of ferroelectric MOC that can really reach the spontaneous polarization status and opens up a new avenue to explore novel MOC-based ferroelectrics. PMID- 19128171 TI - Liquid crystalline corannulene responsive to electric field. AB - An amide-appended corannulene derivative (3) with tribranched paraffinic side chains self-assembles into a hexagonal columnar liquid crystalline (LC) mesophase over a wide temperature range from 154 to -10 degrees C. In contrast with columnar LC assemblies of planar polycyclic aromatic hydrocarbons, the LC assembly of nonplanar 3 responds to an applied electric field and can align homeotropically to the electrode surface. Even after the electric field is switched off, this alignment is memorized for a long period of time unless the material is heated above the clearing temperature. PMID- 19128173 TI - The binary silver nitrogen anion [Ag(N3)2]-. AB - The reaction of silver azide with triphenylsulfonium or tetraphenylphosphonium azide results in the formation of the binary silver-nitrogen compound, the novel diazido-argentate(I) anion [Ag(N(3))(2)](-). The crystal structures of both salts of this anion show different cation dependent azide coordination spheres around silver. PMID- 19128172 TI - Dynamic intracomplex heterogeneity of phytochrome. AB - Low temperature single-molecule fluorescence emission spectroscopy on individual phytochromes from Agrobacterium tumefaciens corroborates findings from ensemble spectroscopy concerning intercomplex heterogeneity. Furthermore, time-dependent intracomplex heterogeneity has been observed. PMID- 19128174 TI - Electronic structure of DNA--unique properties of 8-oxoguanosine. AB - 8-Oxo-7,8-dihydroguanosine (8-oxoG) is among the most common forms of oxidative DNA damage found in human cells. The question of damage recognition by the repair machinery is a long standing one, and it is intriguing to suggest that the mechanism of efficiently locating damage within the entire genome might be related to modulations in the electronic properties of lesions compared to regular bases. Using laser-based methods combined with organizing various oligomers self-assembled monolayers on gold substrates, we show that indeed 8 oxoG has special electronic properties. By using oligomers containing 8-oxoG and guanine bases which were inserted in an all thymine sequences, we were able to determine the energy of the HOMO and LUMO states and the relative density of electronic states below the vacuum level. Specifically, it was found that when 8 oxoG is placed in the oligomer, the HOMO state is at higher energy than in the other oligomers studied. In contrast, the weakly mutagenic 8-oxo-7,8 dihydroadenosine (8-oxoA) has little or no effect on the electronic properties of DNA. PMID- 19128175 TI - Mechanistic insight into the role of transition-state stabilization in cyclophilin A. AB - Peptidyl prolyl cis-trans isomerases (PPIases) are ubiquitous enzymes in biology that catalyze the cis-trans isomerization of the proline imide peptide bond in many cell signaling pathways. The local change of the isomeric state of the prolyl peptide bond acts as a switching mechanism in altering the conformation of proteins. A complete understanding of the mechanism of PPIases is still lacking, and current experimental techniques have not been able to provide a detailed atomistic picture. Here we have carried out several accelerated molecular dynamics simulations with explicit solvent, and we have provided a detailed description of cis-trans isomerization of the free and cyclophilin A-catalyzed process. We show that the catalytic mechanism of cyclophilin is due mainly to the stabilization and preferential binding of the transition state that is achieved by a favorable hydrogen bond interaction with a backbone NH group. We also show that the substrate in the transition state interacts more favorably with the enzyme than the cis isomer, which in turn interacts more favorably than the trans isomer. The stability of the enzyme-substrate complex is directly correlated with the interaction the substrate makes with a highly conserved arginine residue. Finally, we show that catalysis is achieved through the rotation of the carbonyl oxygen on the N-terminal of the prolyl peptide bond in a predominately unidirectional fashion. PMID- 19128176 TI - Surface engineering using Kumada catalyst-transfer polycondensation (KCTP): preparation and structuring of poly(3-hexylthiophene)-based graft copolymer brushes. AB - Poly(4-vinylpyridine)-block-poly(4-iodo-styrene), P4VP-b-PS(I), block copolymers obtained by iodination of readily available P4VP-b-PS block copolymers strongly adhere to variety of polar substrates including Si wafers, glasses, or metal oxide surfaces by a polar P4VP block, forming polymer brushes of moderately stretched PS(I) chains. Kumada catalyst-transfer polycondensation (KCTP) from the P4VP-b-PS(I) brushes results into planar brushes of the graft copolymer in which relatively short ( approximately 10 nm) poly(3-hexylthiophene), P3HT, grafts emanate from the surface-tethered PS(I) chains. Grafting of the P3HT leads to significant stretching of the PS(I) backbone as a result of increased excluded volume interactions. Specific adsorption of the P4VP block to polar surfaces was utilized in this work to pattern the P4VP(25)-b-PS(I)(350) brush. The microscopically structured P4VP(25)-b-PS(I)(350) brush was converted into the respectively patterned P4VP-PS(I)-g-P3HT one using KCTP. We also demonstrated that KCTP from functional block copolymers is an attractive option for nanostructuring with polymer brushes. P4VP(75)-b-PS(I)(313) micelles obtained in selective solvent for the PS(I) block form a quasi-ordered hexagonal array on Si wafer. The P4VP(75)-b-PS(I)(313) monolayer preserves the characteristic quasi regular arrangement of the micelles even after extensive rinsing with various solvents. Although the grafting of P3HT from the nanopatterned P4VP(75)-b PS(I)(313) brush destroys the initial order, the particulate morphology in the resulting film is preserved. We believe that the developed method to structured brushes of conductive polymers can be further exploited in novel stimuli responsive materials, optoectronic devices, and sensors. PMID- 19128177 TI - Measuring aromaticity with the dimethyldihydropyrene ring current probe. Experimental and computational studies of the fulvenes and the strongly antiaromatic cyclopentadienone reveal large Mills-Nixon-type bond localization effects. Synthesis of fulvene-fused dihydropyrenes. AB - The synthesis of the methylfulvene- and phenylfulvene-annelated dihydropyrenes 10 and 22 from the cyclopentadiene-fused dihydropyrene 7 in 68% and 80% yields, respectively, are reported. However, the attempted formation of the parent fulvene-fused dihydropyrene 18 failed, both from the cyclopentadiene 7 with formaldehyde and from the cyclopentadienone 5 in Wittig-type reactions. Chemical shift data for the methylfulvene (35) and phenylfulvene (36)-fused dihydropyrenes 10 and 22 were used to estimate the reduction in the dihydropyrene nucleus aromaticity (DHPN) (relative to benzene fusion) in 10 and 22 (12-16% and 22-25% respectively). Calculations revealed that this reduction in diatropicity, contrary to the situation with benzene fusion, is not due to any aromaticity of the annelating fulvenes but instead is caused by Mills-Nixon-type effects. We conclude that methyl- and phenylfulvene are nonaromatic. An improved synthetic route to the cyclopentadienone 5 was found in an unprecedented cyclization of the trans-cinnamic acid analogue 29 in 80% yield. This enabled an X-ray structure of 5 to be obtained, for comparison to that of the saturated ketone 4. Even though crystals of 5 and 4 show diastereomeric disorder, when the average bond length data of cyclopentadienone 5 is compared with those of cyclopentenone 4 and the parent and benzo dihydropyrenes 6 and 33, it is clearly evident that 5 has the opposite bond-alternation pattern, consistent with a [4n] fused annulene. From the bond length data, cyclopentadienone has approximately 87% of the effect of a benzene ring on bond alternation, which is in reasonable agreement with the previously found NMR value (78%). Structure and nucleus-independent chemical shift calculations support these results. PMID- 19128179 TI - CaFeO2: a new type of layered structure with iron in a distorted square planar coordination. AB - CaFeO(2), a material exhibiting an unprecedented layered structure containing 3d(6) iron in a high-spin distorted square-planar coordination, is reported. The new phase, obtained through a low-temperature reduction procedure using calcium hydride, has been characterized through powder neutron diffraction, synchrotron X ray diffraction, Mossbauer spectroscopy, XAS experiments as well as first principles DFT calculations. The XAS spectra near the Fe-K edge for the whole solid solution (Sr(1-x)Ca(x))FeO(2) supports that iron is in a square-planar coordination for 0 X(3)Sigma(g)(-)) with subcellular spatial resolution. The singlet oxygen lifetime determines its diffusion distance and hence the intracellular volume element in which singlet oxygen-initiated perturbation of the cell occurs. In this study, the time resolved phosphorescence of singlet oxygen produced by the sensitizers chlorin (Chl) and 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) was monitored. These molecules localize in different domains of a living cell. The data indicate that (i) the singlet oxygen lifetime and (ii) the rate constant for singlet oxygen quenching by added NaN(3) depend on whether Chl or TMPyP was the photosensitizer. These observations likely reflect differences in the chemical and physical constituency of a given subcellular domain (e.g., spatially dependent oxygen and NaN(3) diffusion coefficients), thereby providing evidence that singlet oxygen responds to the inherent heterogeneity of a cell. Thus, despite a relatively long intracellular lifetime, singlet oxygen does not diffuse a great distance from its site of production. This is a consequence of an apparent intracellular viscosity that is comparatively large. PMID- 19128182 TI - Factors relevant for the regioselective cyclopolymerization of 1,6-heptadiynes, N,N-dipropargylamines, N,N-dipropargylammonium salts, and dipropargyl ethers by Ru(IV)-alkylidene-based metathesis initiators. AB - The factors relevant for the regioselectivity of insertion of various 1,6 heptadiynes, N,N-dipropargylamines, N,N-dipropargylammonium salts and dipropargyl ethers into different Ru(IV)-alkylidenes, i.e., [Ru(CF(3)COO)(2)(IMesH(2))(CHR), (R = 2,4,5-(MeO)(3)-C(6)H(2) (I1), 2-(2-PrO)-5-NO(2)-C(6)H(3) (I3), 2-(2-PrO) C(6)H(4) (I4)), [Ru(CF(3)COO)(2)(1,3-dimesityl-3,4,5,6-tetrahydropyrimidin-2 ylidene)(CH-2-(2-PrO)-5-NO(2)-C(6)H(3))] (I2), [Ru(CF(3)COO)(2)(3-mesityl-1 ((1'R)-1'-phenylethyl)-imidazolin-2-ylidene)(CH-2-(2-PrO)C(6)H(4))] (I5) and [Ru(C(6)F(5)COO)(2)(IMesH(2))(CH-2-(2-PrO)-C(6)H(4))] (I6), (IMesH(2) = 1,3 dimesitylimidazolin-2-ylidene), is described. (13)C NMR experiments revealed that all polymers synthesized by the action of I1-I6 consisted virtually solely (>95%) of five-membered repeat units, i.e., (cyclopent-1-enylene)-1,2-vinylenes, 3,4-(1H 2,5-dihydropyrrylenium)-3,4-vinylenes, and (2-pentyl-2,5-dihydrofurylene)-3,4 vinylenes, respectively. The (13)C NMR-based assignments were supported by the synthesis of model compounds, i.e., (cyclopent-3-ene-1,1-diyldimethylbis(tris(3,5 dimethoxyphenyl)carboxylate) (MC1) and N-propyl-N-ethyl-2,5-dihydropyrrolium tetrafluoroborate (MC2), as well as by ene-yne cross metathesis reactions of 3 (propargyloxy)-1-octyne (M6) with trimethylallylsilane. In the polymerization of N-ethyl-N,N-dipropargylamine (M9), an intermediate was isolated that sheds light onto the role of heteroatoms in the 4-position of 1,6-heptadiynes in cyclopolymerization. In addition, in the cyclopolymerization of M9 by I4 the product resulting from back-biting has been isolated and explains for the low polymerization propensity of Ru-alkylidenes for N-alkyl-N,N-dipropargylamines. PMID- 19128183 TI - Recovery of both vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus from culture of a single clinical specimen from colonized or infected patients. AB - OBJECTIVE: To describe the incidence of recovery of both vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) from culture of a single clinical specimen, to describe the clinical characteristics of patients from whom these specimens were recovered, and to identify the risk factors of these patients. DESIGN: A retrospective cohort and case-control study. SETTING: A tertiary care university hospital and referral center in Seoul, Korea. METHODS: We identified 61 case patients for whom a single clinical specimen yielded both VRE and MRSA on culture, and 122 control patients for whom any clinical specimen yielded only VRE on culture. The control patients were selected by matching 2:1 with the case patients for age, sex, and first date of sampling that led to isolation of VRE or both VRE and MRSA among 1,536 VRE-colonized patients from January 1, 2003, through December 31, 2006. To identify patient risk factors for the recovery of both VRE and MRSA in a single clinical specimen, we performed univariate comparisons between the 2 groups and then multivariate logistic regression analysis. RESULTS: The incidence of recovery of both VRE and MRSA from culture of a single clinical specimen was 3.97% (for 61 of 1,536 VRE colonized patients) over 4 years. Among these 82 single clinical specimens, the most common type was wound specimens (26.8%), followed by lower respiratory tract specimens (18.3%), urine specimens (17.1%), and catheter tips (15.9%). Of the 61 case patients, 14 (23.0%) had 2 or more single clinical specimens that yielded both VRE and MRSA on culture, and the longest interval from the first sampling that yielded both organisms to the last sampling that yielded both was 174 days. Independent patient risk factors for the presence of both VRE and MRSA in a single clinical specimen were chronic renal disease (odds ratio [OR], 7.00; P=.012 ), urinary catheterization (OR, 3.36; P=.026), and longer total cumulative duration of hospital stay within the previous year (OR, 1.03; P < .001). CONCLUSION: We confirmed that the recovery of VRE and MRSA from a single clinical specimen occurs continually. Because prolonged cell-to-cell contact can facilitate transfer of vanA, close observation and surveillance for vancomycin resistant S. aureus, especially among patients with risk factors for the recovery of both VRE and MRSA from a single clinical specimen, should be continued. PMID- 19128184 TI - Emergence of community-associated methicillin-resistant Staphylococcus aureus strains as a cause of healthcare-associated bloodstream infections in Korea. AB - BACKGROUND: The prevalence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains causing bloodstream infection (BSI) has not been studied in Korea. OBJECTIVE: We sought to determine the prevalence of CA MRSA strains among isolates recovered from patients with MRSA BSIs and to explore epidemiological changes in Korea. We also sought to evaluate clinical characteristics relevant to the development of healthcare-associated BSIs. METHODS: We prospectively collected consecutive MRSA isolates from patients with BSI at 4 hospitals from July 1 through November 30, 2007, and we also included MRSA isolates recovered from culture of blood samples collected during a previous year (October 1, 2004 through September 30, 2005) at a different hospital. Molecular typing studies were performed, including pulsed-field gel electrophoresis (PFGE), multilocus sequence typing, Staphylococcus protein A (spa) typing, and staphylococcal cassette chromosome mec (SCCmec) typing. We compared the clinical characteristics and outcomes of patients with healthcare associated BSI due to CA-MRSA strains with those of patients with healthcare associated BSI due to healthcare-associated MRSA (HA-MRSA) strains. RESULTS: There were 76 cases of MRSA BSI, of which 4 (5.3%) were community-associated and 72 (94.7%) were healthcare-associated. Among the 72 HA-MRSA BSIs, 18 (25%) were community onset, and 54 (75%) were hospital onset. PFGE type D-ST72-spa B-SCCmec type IVA MRSA, the predominant genotype of CA-MRSA in Korea, accounted for 19 (25%) of all 76 MRSA BSIs, including 17 (23.6%) of 72 HA-MRSA BSIs and 11 (20.8%) of 53 hospital-onset HA-MRSA BSIs. Patients with healthcare-associated BSIs due to CA-MRSA strains carrying SCCmec type IVA tended to have fewer healthcare associated risk factors, compared with patients with healthcare-associated BSIs due to HA-MRSA strains carrying other SCCmec types. The presence of a central venous catheter or other invasive device was the only independent factor differentiating patients infected with hospital-associated genotype strains from patients infected with other strains. Clinical outcomes were similar between both groups. CONCLUSIONS: CA-MRSA strains are emerging as a major cause of BSI in healthcare settings in Korea. This changing epidemiology of MRSA poses a challenge to public health and infection control in hospital settings. PMID- 19128185 TI - Adsorption and reactivity of CO(2) on defective graphene sheets. AB - Density-functional calculations have been performed to investigate the adsorption of CO(2) on defected graphite (0001) represented by a single graphene sheet. The interaction with a vacancy defect gives a computed molecular binding energy of approximately 136 meV in a strong physisorbed state. Subsequently, chemisorption by lactone group formation will occur after overcoming a barrier of approximately 1 eV relative to the gas phase, with an exothermicity of about 1.4 eV. Further reaction paths from this chemisorbed state lead to dissociation of the CO(2) through the formation of epoxy groups followed by oxygen recombination and desorption of O(2), after overcoming successive energy barriers of approximately 0.9 and approximately 1.0 eV. The global minimum ("O(2) desorbed + graphene sheet") entails an energy release of about 3.4 eV with respect to the initial state. PMID- 19128186 TI - Proton affinity and zwitterion stability: new results from infrared spectroscopy and theory of cationized lysine and analogues in the gas phase. AB - The gas-phase structures of alkali metal cationized lysine (Lys), alpha-N methyllysine (NMeLys), and epsilon-N,N-dimethyllysine (Lys(Me)(2)) are investigated using infrared multiple photon dissociation (IRMPD) spectroscopy utilizing light generated by a free electron laser and ab initio calculations. The proton affinities of the compounds span a range of approximately 20 kJ/mol. For NMeLys x M(+), experiment and theory indicate that NMeLys is nonzwitterionic for M = Li and zwitterionic for M = Na and K. For Lys(Me)(2) x M(+), experiment and theory indicate that Lys(Me)(2) is zwitterionic for M = Li, Na, and K. This is the first spectroscopic observation of the zwitterionic form of an amino acid complexed with Li(+). The results are compared with IRMPD spectra reported previously for Lys and -N-methyllysine (Lys(Me)) complexed with Li, Na, and K, and new calculations performed at higher levels of theory for those ions. The combined experimental and theoretical results indicate that protonation in the zwitterionic forms of the these amino acids is favored at the more basic methylated amine site, but that any relationship between the proton affinity of the amino acid and the relative zwitterion stability of the alkali metal cationized amino acid is only indirect. These results provide additional evidence that proton affinities are not a reliable indicator of zwitterion stability for cationized amino acids because side chains can have very different effects on the stability of different conformers in the neutral, protonated, and metal cationized forms. PMID- 19128187 TI - Making imines without making water-exploiting a recognition-mediated aza-wittig reaction. AB - A recognition-mediated aza-Wittig reaction permits the efficient formation of an imine in dry CDCl(3) from an iminophosphorane and an aldehyde. The kinetic and thermodynamic parameters for this reaction are compared with those obtained from a condensation reaction between an aldehyde and an amine that forms the same product. PMID- 19128188 TI - Enantioselective, ketoreductase-based entry into pharmaceutical building blocks: ethanol as tunable nicotinamide reductant. AB - The use of NADH- and NADPH-dependent ketoreductases to access enantioenriched pharmaceutical building blocks is reported. Seven structurally diverse synthons are obtained, including those for atomoxetine (KRED 132), talampanel (RS1-ADH and CPADH), Dolastatin (KRED 132), and fluoxetine (KRED 108/132). Ethanol may be used as stoichiometric reductant, regenerating both nicotinamide cofactors, particularly under four-electron redox conditions. Its favorable thermodynamic and economic profile, coupled with its advantageous dual cosolvent role, suggests a new application for biomass-derived ethanol. PMID- 19128189 TI - Ni(0)-catalyzed conjugate addition of Me3SiCN to ynones: alpha-bromo-beta-cyano tetrasubstituted enones. AB - Conjugate addition of Me(3)SiCN to ynones is smoothly catalyzed by Ni(cod)(2) to give the beta-cyanosilyloxyallene quantitatively. Further reaction of the silyloxyallenes with NBS provides the tetrasubstituted alpha-bromo-beta-cyano enones in high yields (up to 95%) with excellent Z-selectivity (E/Z = up to >1/99). X-ray crystallographic analysis showed a bent structure of the alpha bromo-beta-cyano enone due to a deconjugation of the pi-bond and carbonyl group. PMID- 19128190 TI - Strict reagent control in the asymmetric allylboration of N-TIPS-alpha-amino aldehydes with the B-allyl-10-TMS-9-borabicyclo[3.3.2]decanes. AB - The allylboration of enantiomerically pure N-triisopropylsilyl-alpha-amino aldehydes (2) with B-allyl-10-trimethylsilyl-9-borabicyclo[3.3.2]decanes (1) proceeds cleanly at -78 degrees C, exhibiting essentially complete reagent control. After an oxidative workup, an HOAc-mediated N-->O TIPS rearrangement facilitates the clean formation of stable O-TIPS protected beta-amino alcohol derivatives 3 which are isolated in 60-83% yields in > or = 96% de and > 99% ee. For the leucinal series (R = i-Bu), an efficient entry to either statine (8aSS) or epi-statine (8aRS) is reported illustrating the versatility of this potent 1/2 combination. PMID- 19128192 TI - A facile deprotection of secondary acetamides. AB - Imidoyl chlorides, generated from secondary acetamides and oxalyl chloride, can be harnessed for a selective and practical deprotection sequence. Treatment of these intermediates with 2 equiv of propylene glycol and warming enables the rapid release of amine hydrochloride salts in good yields. Notably, the reaction conditions are mild enough to allow for a swift deprotection with no observed epimerization of the amino center. PMID- 19128191 TI - Attempts to improve the overall stereoselectivity of the Ireland-Claisen rearrangement. AB - With focus on the steric effects present in the transition states for the [3,3] sigmatropic rearrangement, the substrate 5 has been designed to improve the overall stereoselectivity of the Ireland-Claisen rearrangement. Experimentally, it has been found that (1) only Z-6 rearranges to 7 at 80 degrees C and (2) E-6 isomerizes to Z-6 at 80 degrees C, thereby allowing the transformation of 5 into 7 in an almost quantitative yield. To illustrate the usefulness of this approach, two additional examples are given. PMID- 19128193 TI - Enantioselective zirconium-catalyzed Friedel-Crafts alkylation of pyrrole with trifluoromethyl ketones. AB - The first catalytic enantioselective Friedel-Crafts alkylation of pyrrole with 2,2,2-trifluoroacetophenones to give pyrroles with a trifluoromethyl-substituted tertiary alcohol moiety bearing a quaternary stereogenic center is described. The reaction is achieved in the presence of a 3,3'-dibromo-BINOL-Zr(IV) complex to give the expected products with high yields (up to 98%) and good enantioselectivities (up to 93% ee). The absolute stereochemistry of the products has been determined by chemical correlation. PMID- 19128194 TI - FeCl3-catalyzed 1,2-addition reactions of aryl aldehydes with arylboronic acids. AB - A novel protocol for the 1,2-addition reactions of electron-deficient aryl aldehydes with arylboronic acids using an inexpensive and environmentally benign iron catalyst is reported. In the presence of FeCl(3) and 2-(di-tert butylphosphino)biphenyl, 1,2-addition reactions of various electron-deficient aryl aldehydes with arylboronic acids provided the corresponding biaryl methanols in moderate to excellent yields. PMID- 19128195 TI - Exploration of the "traceless" reductive ligation of S-nitrosothiols. AB - The first "traceless" reductive ligation of S-nitrosothiols using phosphine ester/thioester conjugates is reported. Experiments also show that stable thioimidate compounds could be formed in the reaction between S-nitrosothiols and some phosphine-thioester substrates. PMID- 19128198 TI - Drug addiction and brain targets: from preclinical research to pharmacotherapy. PMID- 19128200 TI - Selective antagonism at dopamine D3 receptors as a target for drug addiction pharmacotherapy: a review of preclinical evidence. AB - The focal distribution of the dopamine (DA) D(3) receptor in brain regions implicated in emotional and cognitive functions has made this target a main focus of drug discovery efforts. This paper will review the most recent lines of research in support of the use of selective DA D(3) receptor antagonists for the pharmacotherapeutic management of drug addiction: (1) expression of the DA D(3) receptor in the rodent and human brain; (2) changes in expression of the DA D(3) receptor following exposure to drugs of abuse, and (3) efficacy of selective DA D(3) receptor antagonists in preclinical paradigms assessing the behavioral effects of drugs such as cocaine, nicotine, alcohol, methamphetamine, and heroin. This manuscript, however, will not review the effects of nonselective DA D(2)/D(3) receptor antagonists or partial D(3) receptor agonists. Growing evidence suggests that selective DA D(3) receptor antagonists do not affect the primary reinforcing effects of drugs of abuse, but rather seem to regulate the motivation to self-administer drugs under schedules of reinforcement that require an increase in work demand. In addition, selective antagonism at DA D(3) receptors appears to disrupt significantly the responsiveness to drug-associated stimuli that play a key role in reinstatement of drug-seeking behavior. These preclinical findings will be discussed in the context of translational research relevant to the design of early clinical trials and hypothesis testing in humans. PMID- 19128201 TI - Neuronal nicotinic receptors as brain targets for pharmacotherapy of drug addiction. AB - Nicotine addiction and other forms of drug addiction continue to be significant public health problems in the United States and the rest of the world. Accumulated evidence indicates that brain nicotinic acetylcholine receptors (nAChRs) are a heterogenous family of ion channels expressed in the various parts of the brain. A growing body of preclinical studies suggests that brain nAChRs are critical targets for the development of pharmacotherapies for nicotine and other drug addictions. In this review, we will discuss the nAChR subtypes, their function in response to endogenous brain transmitters, and how their functions are regulated in the presence of nicotine. Furthermore, we will discuss the role of nAChRs in mediating nicotine-induced addictive behavior in animal models. Additionally, we will provide an overview of the effects of nicotine and nicotinic compounds on the mesolimbic dopamine system, part of the reinforcement/reward circuitry of the brain, as an example of the neurochemical basis of nicotine addiction and other drug addictions. An appreciation of the complexity of nicotinic receptors and their regulation will be necessary for the development of nicotinic receptor modulators as potential pharmacotherapy for drug addiction. PMID- 19128199 TI - Role of the dopamine transporter in the action of psychostimulants, nicotine, and other drugs of abuse. AB - A number of studies over the last two decades have demonstrated the critical importance of dopamine (DA) in the behavioral pharmacology and addictive properties of abused drugs. The DA transporter (DAT) is a major target for drugs of abuse in the category of psychostimulants, and for methylphenidate (MPH), a drug used for treating attention deficit hyperactivity disorder (ADHD), which can also be a psychostimulant drug of abuse. Other drugs of abuse such as nicotine, ethanol, heroin and morphine interact with the DAT in more indirect ways. Despite the different ways in which drugs of abuse can affect DAT function, one evolving theme in all cases is regulation of the DAT at the level of surface expression. DAT function is dynamically regulated by multiple intracellular and extracellular signaling pathways and several protein-protein interactions. In addition, DAT expression is regulated through the removal (internalization) and recycling of the protein from the cell surface. Furthermore, recent studies have demonstrated that individual differences in response to novel environments and psychostimulants can be predicted based on individual basal functional DAT expression. Although current knowledge of multiple factors regulating DAT activity has greatly expanded, many aspects of this regulation remain to be elucidated; these data will enable efforts to identify drugs that might be used therapeutically for drug dependence therapeutics. PMID- 19128202 TI - Targeting endogenous mu- and delta-opioid receptor systems for the treatment of drug addiction. AB - Drug addiction is a chronic, relapsing disorder that is characterized by a compulsion to take drug regardless of the adverse consequences that may ensue. Although the involvement of mesoaccumbal dopamine neurons in the initiation of drug abuse is well-established, neuroadaptations within the limbic cortical- striatopallidal circuit that occur as a consequence of repeated drug use are thought to lead to the behavioral dysregulation that characterizes addiction. Opioid receptors and their endogenous ligands are enriched in brain regions comprising this system and are, thus, strategically located to modulate neurotransmission therein. This article will review data suggesting an important role of mu-opioid receptor (MOPr) and delta opioid receptor (DOPr) systems in mediating the rewarding effects of several classes of abused drugs and that aberrant activity of these opioid systems may not only contribute to the behavioral dysregulation that characterizes addiction but to individual differences in addiction vulnerability. PMID- 19128205 TI - Glutamate: the new frontier in pharmacotherapy for cocaine addiction. AB - Considerable research into the neurobiology of cocaine addiction has shed light on the role of glutamate. Findings from models of relapse to cocaine-seeking indicate that the glutamatergic system is critically involved, as glutamate levels in the nucleus accumbens increase during reinstatement and glutamate receptor activation is necessary for reinstatement to drug-seeking. Thus, it would seem beneficial to block the increased glutamate release, but full antagonists of ionotropic glutamate receptors produce undesirable side effects. Therefore, modulation of glutamatergic transmission would be advantageous and provide novel pharmacotherapeutic avenues. Pharmacotherapies have been developed that have the potential to modulate excessive glutamatergic transmission through ionotropic and metabotropic (mGluR) glutamate receptors. Compounds that modulate glutamatergic transmission through ionotropic glutamate receptors include the non competitive N-methyl-D-aspartic acid antagonists, amantadine and memantine, and the partial N-methyl-D-aspartic acid agonist d-cycloserine. They have shown promise in preclinical models of cocaine addiction. The mGluR2/3 agonist LY379268 is effective in inhibiting cocaine seeking in preclinical animal models and could decrease stress-induced relapse due to its anxiolytic effects. Similarly, the mGluR1/5 antagonists, 2-methyl-6-(phenylethynyl)pyridine and 3-[2-methyl-4 thiazolyl)ethynyl]pyridine, have shown to be effective in preclinical models of cocaine addiction. The cysteine pro-drug, N-acetylcysteine, restores the inhibitory tone on presynaptic glutamate receptors and has been effective in reducing cue-induced craving and cocaine use in humans. Furthermore, anticonvulsants, such as topiramate or lamotrigine, have shown efficacy in treating cocaine dependence or reducing relapse in humans. Future pharmacotherapy may focus on manipulating signal transduction proteins and pathways, which include Homer/N-methyl-D-aspartic acid complexes, to provide effective treatment for cocaine addiction. PMID- 19128206 TI - Alzheimer's disease, ceramide, visfatin and NAD. AB - This review discusses new mechanisms for the induction of Alzheimer's disease, involving lipid toxicity and adipokines. Ceramide induces oxidative stress and the formation of amyloid beta. Visfatin induces oxidative stress, damages the blood brain barrier and increases the attraction of monocytes, neutrophils and other white blood cells. A new mechanism for visfatin/NAD (nicotinamide adenine dinucleotide)-induced oxidative stress is presented involving redox cycling catalyzed by xanthine dehydrogenase and NADH oxidase. These mechanisms are discussed in terms of the treatment of Alzheimer's disease. PMID- 19128203 TI - The role of 5-HT3 receptors in drug abuse and as a target for pharmacotherapy. AB - Alcohol and drug abuse continue to be a major public health problem in the United States and other industrialized nations. Extensive preclinical research indicates the mesolimbic dopamine (DA) pathway and associated regions mediate the rewarding and reinforcing effects of drugs of abuse and natural rewards, such as food and sex. The serotonergic (5-HT) system, in concert with others neurotransmitter systems, plays a key role in modulating neuronal systems within the mesolimbic pathway. A substantial portion of this modulation is mediated by activity at the 5-HT3 receptor. The 5-HT3 receptor is unique among the 5-HT receptors in that it directly gates an ion channel inducing rapid depolarization that, in turn, causes the release of neurotransmitters and/or peptides. Preclinical findings indicate that antagonism of the 5-HT3 receptor in the ventral tegmental area, nucleus accumbens or amygdala reduces alcohol self-administration and/or alcohol associated effects. Less is known about the effects of 5-HT3 receptor activity on the self-administration of other drugs of abuse or their associated effects. Clinical findings parallel the preclinical findings such that antagonism of the 5 HT3 receptor reduces alcohol consumption and some of its subjective effects. This review provides an overview of the structure, function, and pharmacology of 5-HT3 receptors, the role of these receptors in regulating DA neurotransmission in mesolimbic brain areas, and discusses data from animal and human studies implicating 5-HT3 receptors as targets for the development of new pharmacological agents to treat addictions. PMID- 19128208 TI - The proNGF-p75NTR-sortilin signalling complex as new target for the therapeutic treatment of Parkinson's disease. AB - Growing evidence has shown that the p75 neurotrophin receptor (p75NTR) may play important roles in controlling neuronal survival or cell apoptosis within the central nervous system in development, and in pathological or neural injury. Recent studies have further revealed that p75NTR acts as a "molecular signal switch" that determines cell death or survival by three processes. First, pro nerve growth factor (proNGF) triggers cell apoptosis by its high affinity binding to p75NTR, while NGF induces neuronal survival with low-affinity binding. Second, p75NTR mediates cell death by combining with co-receptor sortilin, whereas it promotes neuronal survival through combination with proNGF. Third, release of the intracellular domain chopper or cleavaged "short p75NTR" can independently initiate neuronal apoptosis. We have identified the cell self-destructive proNGF p75NTR-sortilin signalling apparatus assembled in ventral tier dopamine neurons of the substantia nigra pars compacta, suggesting that p75NTR signalling might be involved in selective cell death mechanisms of substantia nigra neurons or disease progression of Parkinson's disease (PD). In addition, experimental manipulation of p75NTR benefited cell survival of cholinergic or motor neurons and improved disease progression of the neurodegenerative diseases Alzheimer's disease and Amyotrophic lateral sclerosis. The proNGF-p75NTR-sortilin signalling complex may thus provide new target for neuroprotection of substantia nigra neurons and the therapeutic treatment of PD. PMID- 19128207 TI - Alzheimer's disease drug development: old problems require new priorities. AB - Alzheimer's disease (AD) clinical drug development and patient care depend on rating instruments, research designs and methods, and translations of clinical trial (CT) results into the clinic without support from standardized protocols able to control (i) random measurement error intrusions into observations, (ii) inaccuracy and bias introduced by clinical evaluators, (iii) conformity of research sites to conditions of research protocols, (iv) the ability of the CT to model for practitioners the most effective use of the drug with individual patients, and (v) other factors able to invalidate research and patient care data. This relaxed attitude with regard to AD methods may be changing now with Alzheimer's Disease Neuroimaging Initiative (ADNI) evidence that carefully standardized protocols are needed to validate biomarkers for use in AD diagnosis, drug development, and patient care. In the fields of psychiatry and AD, recent studies have detected serious inaccuracies, imprecision, biases and compromises of study protocols able to invalidate CT outcome data and conclusions drawn from these data. This limited but troubling evidence reinforces ADNI calls for more detailed methodological protocols. Based on the limits to precision and accuracy associated with rated outcomes in CTs and patient care, we call for priority to be given to the qualification and use of biomarkers as outcome variables in AD drug development and patient care and, to insure effective uses of biomarkers, to development of protocol guided practices being modeled in ADNI research. To meet clinical pharmacology's therapeutic aims we conclude that AD CTs need to set for clinicians the conditions of use of drugs shown efficacious, biomarker surrogate endpoints as drug targets, and not to function merely as tests for efficacy conducted under field conditions determined by current clinical practices. PMID- 19128209 TI - Central sensitisation phenomena in primary headaches: overview of a preventive therapeutic approach. AB - Central sensitisation phenomena have been well recognized in the development of migraine attacks and tension type headache. It is also known that headache frequency is related to sensitization. Though some studies have focused on the effects of symptomatic treatment on allodynia, few reports have described the action of preventive agents on the facilitating factors for central sensitisation. In this study we aim to review the factors concurrent with an increase in central sensitisation, in view of the choice of preventive agents for primary headaches. Central sensitisation phenomena are increased in pain syndromes with psycho-pathological co-morbidities. For instance, sleep disorders are a frequent symptom in headache, prevailing in chronic forms and in patients with psychiatric comorbidity. Sleep deprivation is also a factor producing hyperalgesic changes. It is known that symptoms attributable to central sensitization are diffusely pronounced in fibromyalgic (FMS) patients, and that FMS co-morbidity is frequent in primary headaches and associated with higher frequency and poorer quality of life. We report our preliminary experience in a group of 20 chronic migraine patients, treated with duloxetine 60 mg/die vs a self-management program including stretching (relaxation training) and exercise (cervical-dorsal flexion and rotation) to decrease strength and flexibility of muscles of cervical and dorsal spine headache patients. Both the treatments were effective on headache frequency and pericranial tenderness, although FMS comorbidity significantly reduced their efficacy on migraine and quality of life. The whole spectrum of action of pharmacological and non pharmacological treatments on central sensitisation mechanisms, and on their facilitating factors, should be taken into account for the best preventive therapeutic approach of primary headaches. PMID- 19128210 TI - NF-kappa B, a potential therapeutic target for the treatment of multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS) that afflicts over 2 million people worldwide. On the basis of the temporal course of disease, MS can be subdivided into three clinical groups: relapsing remitting MS (RR-MS), secondary progressive MS and primary progressive MS. There is a high degree of clinical diversity within these subgroups. The pathogenesis of MS in most patients is likely to result from autoreactive, activated CD4(+) T cells moving from the periphery across the blood brain barrier into the CNS. Most therapeutic agents used in MS (e.g. immunosuppressive and immunomodulatory drugs and cell cycle interruption drugs) are only used for RR-MS. These treatments show some efficiency in lessening the relapse rate in RR-MS and time to progression, but cannot cure MS. Thus, there is a need for new efficient treatments for all types of MS. An increasing number of studies indicate that nuclear factor-kappaB plays an important role in controlling expression of genes relevant to the pathogenesis of autoimmunity. Genetic factors related to NF-kappaB may also be determinants of MS susceptibility, as polymorphisms in the molecules involved in regulation of the NF-kappaB signal transduction pathway differ between RR-MS and progressive MS. Herein, the role of NF-kappaB in MS will be reviewed and its potential as a new therapeutic target in MS will be considered and compared with existing treatments. PMID- 19128211 TI - Protein-drug interactions. PMID- 19128204 TI - The endocannabinoid system: a new molecular target for the treatment of tobacco addiction. AB - Tobacco addiction is one of the leading preventable causes of mortality in the world and nicotine appears to be the main critical psychoactive component in establishing and maintaining tobacco dependence. Several lines of evidence suggest that the rewarding effects of nicotine, which underlie its abuse potential, can be modulated by manipulating the endocannabinoid system. For example, pharmacological blockade or genetic deletion of cannabinoid CB(1) receptors reduces or eliminates many behavioral and neurochemical effects of nicotine that are related to its addictive potential. This review will focus on the recently published literature about the role of the endocannabinoid system in nicotine addiction and on the endocannabinoid system as a novel molecular target for the discovery of medications for tobacco dependence. PMID- 19128213 TI - Molecular docking algorithms. AB - By means of virtual screening of small molecules databases it is possible to identify new potential inhibitors against a target of interest. Molecular docking is a computer simulation procedure to predict the conformation of a receptor ligand complex. Each docking program makes use of one or more specific search algorithms, which are the methods used to predict the possible conformations of a binary complex. In the present review we describe several molecular-docking search algorithms, and the programs which apply such methodologies. We also discuss how virtual screening can be optimized, describing methods that may increase accuracy of the simulation process, with relatively fast docking algorithms. PMID- 19128212 TI - Computational methods for calculation of ligand-binding affinity. AB - Precise computational methods to determine ligand-binding affinity are needed to accelerate the discovery of new drugs. Assessing protein-ligand interaction is of great importance for virtual screening initiatives. The affinity may be computational evaluated using scoring functions involving terms for intermolecular hydrogen bonds, contact surface, hydrophobic contacts, electrostatic interactions and others. Empirical scoring functions have been developed to evaluate ligand-binding affinity very rapidly. In addition to predict affinity, these scoring functions have been employed to identify the best results obtained from docking simulations. This review describes several computational methods, employed to estimate ligand-binding affinity and discuss their development and main applications. PMID- 19128214 TI - Protein crystallography in drug discovery. AB - Protein crystallography is the main technique used to obtain three-dimensional information for binary complexes involving protein and drugs. Once a protein target has its three-dimensional structure elucidated, the next natural step is the solving of the structure complexed either with its natural substrate, or any ligand or even an inhibitor. Such information is of pivotal importance to understand the structural basis for inhibition of an enzyme. The relevant features, for application of protein crystallography to drug discovery, are discussed in this review. PMID- 19128215 TI - In silico and in vitro: identifying new drugs. AB - Drug development is a high cost and laborious process, requiring a number of tests until a drug is made available in the market. Therefore, the use of methods to screen large number of molecules with less cost is crucial for faster identification of hits and leads. One strategy to identify drug-like molecules is the search for molecules able to interfere with a protein function, since protein interactions control most biological processes. Ideally the use of in silico screenings would make drug development faster and less expensive. Currently, however, the confirmation of biological activity is still needed. Due to the complexity of the task of drug discovery, an integrated and multi-disciplinary approach is ultimately required. Here we discuss examples of drugs developed through a combination of in silico and in vitro strategies. The potential use of these methodologies for the identification of active compounds as well as for early toxicity and bioavailability is also reviewed. PMID- 19128216 TI - Evaluation of molecular docking using polynomial empirical scoring functions. AB - Molecular docking simulations are of pivotal importance for analysis of protein ligand interactions and also an essential resource for virtual-screening initiatives. In molecular docking simulations several possible docked structures are generated, which create an ensemble of structures representing binary complexes. Therefore, it is crucial to find the best solution for the simulation. One approach to this problem is to employ empirical scoring function to identify the best docked structure. It is expected that scoring functions show a descriptive funnel-shaped energy surface without many false minima to impair the efficiency of conformational sampling. We employed this methodology against a test set with 300 docked structures. Docking simulations of these ligands against enzyme binding pocket indicated a funnel-shaped behavior of the complexation for this system. This review compares a set of recently proposed polynomial empirical scoring functions, implemented in a program called POLSCORE, with two popular scoring function programs (XSCORE and DrugScore). Overall comparison indicated that POLSCORE works better to predict the correct docked position, for the ensemble of docked structures analyzed in the present work. PMID- 19128217 TI - Experimental approaches to evaluate the thermodynamics of protein-drug interactions. AB - Precise experimental methods to determine ligand-binding affinity are needed to accelerate the discovery of new drugs. Assessing protein-ligand interaction is of great importance for drug development. One of the techniques that may be used to evaluate ligand-binding affinitty is isothermal titration calorimetry (ITC). This experimental methodology may be used to measure the heat of binding of a ligand to a protein. Furthermore, the development of new empirical scoring functions to assess evaluation protein-ligand interaction lack abundance of experimental information to be used to generate reliable scores. ITC technique may be used to fill this gap. Here we describe the application of this technique to ligand binding affinity determination, and discuss the synergetic relationship between ITC data and the development of a new generation of empirical scoring functions. PMID- 19128218 TI - Molecular recognition models: a challenge to overcome. AB - Molecular recognition process describes the interaction involving two molecules. In the case of biomolecules, these pairs of molecules could be protein-protein, protein-ligand or protein-nucleic acid. The first model to capture the essential features, behind the molecular recognition problem, was the lock-and-key paradigm. The overall analysis protein-protein, protein-nucleic acid and protein ligand interaction based on the three-dimensional structures and physicochemical parameters, such as binding affinity, opened the possibility to provide further insights in this basic phenomenon. The main ideas behind the molecular recognition are discussed in the present review. PMID- 19128219 TI - Molecular modeling as a tool for drug discovery. AB - With the progression of structural genomics projects, comparative modeling remains an increasingly important method of choice to obtain 3D structure of proteins. It helps to bridge the gap between the available sequence and structure information by providing reliable and accurate protein models. Comparative modeling based on more than 30% sequence identity is now approaching its natural template-based limits and further improvements require the development of effective refinement techniques capable of driving models toward native structure. For difficult targets, for which the most significant progress in recent years has been observed, optimal template selection and alignment accuracy are still the major problems. The past year has seen a maturation of molecular modeling, with an increasing number of comparative studies between established methods becoming possible, together with an explosion of new works especially in the areas of combinatorial chemistry and molecular diversity. To achieve this, knowledge about three-dimensional protein structures is crucial for the understanding of their functional mechanisms, and for a rational drug design. This review described recent progress in molecular modeling methodology. PMID- 19128220 TI - Drug-binding databases. AB - Recent developments in computer power and chemoinformatics methodology make possible that a huge amount of data become available through internet. These databases are devoted to a wide spectrum of scientific fields. Here we are concerned with databases related to protein-drug interactions. More specifically, databases where potential new molecules could be accessed to be used in virtual screening initiatives. In the past decade several databases have been developed where molecules to be used in the virtual screening could be easily identified, downloaded and even purchased. This review describes and summarizes the recent advances in the development of these databases, and also the main applications related to virtual screening projects. PMID- 19128221 TI - Linear interaction energy (LIE) method in lead discovery and optimization. AB - Currently, in order to accelerate the process of drug development and also reduce costs, many of the experimental assays related to lead discovery and lead optimization processes are being replaced by computational, in silico, methods. In this context, the LIE (linear interaction energy) method has been used to calculate binding free energies for widely different compounds by averaging interaction energies obtained from molecular dynamics (MD) or Monte Carlo (MC) simulations. In particular, the combination of docking and affinity predictions with the LIE method can thus save valuable resources in lead discovery and optimization projects. This review presents a description of LIE methodology and some recent studies that illustrate the importance and utility of the method in the field of pharmaceutical research. PMID- 19128222 TI - Drugs targeted to improve endothelial function: clinical correlates between sexual and internal medicine. PMID- 19128223 TI - The triad: erectile dysfunction--endothelial dysfunction--cardiovascular disease. AB - Endothelial dysfunction is an important process in the development of atherosclerotic cardiovascular disease, while it is also a major pathophysiological mechanism underlying vasculogenic erectile dysfunction (ED). Expectedly, these two prevalent disorders are linked also at the clinical level: ED is common in patients with overt and silent coronary artery disease, while ED is increasingly being regarded as the early clinical manifestation of a generalized vascular disease and carries an independent risk for future cardiovascular events. The emerging awareness of ED as a barometer for cardiovascular disease offers a unique opportunity to enhance preventive vascular health in men. Lifestyle and risk factor modification, as well as pharmacologic therapy (both phosphodiesterase type-5 inhibitors and non-ED-targeting drugs), appear to confer additional benefit both in terms of ED treatment and overall cardiovascular risk; this benefit may be related, at least partly, to the improvement of endothelial function and anti-inflammatory effects. The present review identifies pathophysiologic links between endothelial dysfunction, ED and coronary artery disease, presents methodological aspects regarding penile and systemic endothelial function, and discusses the clinical implications in terms of diagnosis of ED, assessment of patient risk, and treatment. PMID- 19128224 TI - Non-invasive diagnostic tools for investigating endothelial dysfunction. AB - The endothelium is not merely a barrier but it plays a key role in the maintenance of vascular homeostasis. A dysfunctional endothelium is an early marker of the development of atherosclerotic changes and can also contribute to cardiovascular events. Vascular reactivity tests represent the most widely used methods in the clinical assessment of endothelial function and in the last two decades, several methodologies were developed to study it non invasively in the peripheral macrocirculation (conduit arteries) and microcirculation (resistance arteries and arterioles). This review will centre on the most relevant available non-invasive techniques in the research on endothelial function, their advantages and limitations. Flow mediated dilation (FMD) of the brachial artery by ultrasounds is the most widely used vascular test to assess endothelium-dependent vasodilation. Other approaches include measurement of microcirculatory reactive hyperaemia by forearm venous pletysmography or digital pulse amplitude tonometry, response to beta2 agonist by applanation tonometry or digital photoplethysmography and several test by skin laser doppler. It appears that FMD is the most reproducible test when an appropriate and accurate methodology is applied. Recently, post-ischemic vasodilation in the cavernous arteries was also suggested to study endothelial function in patients with erectile dysfunction. Systemic markers proposed as measures of NO biology, inflammatory cytokines, adhesion molecules, or markers of endothelial damage and repair have only a very limited role as a result of biological and assay availability and variability, these factors currently have a limited role in the assessment of individual patients. The optimal methodology for investigating the multifaceted aspects of endothelial dysfunction is still under debate. Therefore, no available test to assess endothelial function has sufficient sensitivity and specificity to be used yet in clinical practice. Only the growing concordant results from different reproducible and reliable non-invasive methods exploring endothelial function with different stimuli will support and strengthen experimental findings, thus providing conclusive answers in this area of research. PMID- 19128225 TI - The brain, the penis and steroid hormones: clinical correlates with endothelial dysfunction. AB - Erectile function is a complex neurovascular process that depends on the health of the central and peripheral nervous systems and the vasculature. Thus, signaling from the central nervous system (brain) to the peripheral nervous system (penis) is critical and is modulated by a set of complex interactions that depend on cerebral and vascular circulation. The cerebral and peripheral vasculatures are target tissues for sex steroid hormones. Gonadal, adrenal and neurosteroids regulate the function and physiology of the endothelium and modulate vascular and cerebral circulation by genomic and non-genomic dependent mechanisms. Recent advances in cell and molecular biology have defined a critical role of endothelium in vascular function. A host of biochemical and clinical markers of endothelium function and dysfunction have been identified to assess vascular pathology. Emerging evidence suggests that sex steroid hormones play an important role in maintaining endothelial health and sex steroid deficiency is associated with endothelial dysfunction, vascular disease and erectile dysfunction. Such information has important clinical implications in patient management with sex steroid hormone insufficiency, diabetes, metabolic syndrome, vascular disease and erectile dysfunction. In this review, we discuss the role of sex steroid hormones in modulation of the biochemical and clinical markers associated with endothelial dysfunction. Specifically the regulation of endothelial nitric oxide synthase, asymmetric dimethylarginine, reactive oxygen species, endothelin-1, inflammatory cytokines, tumor necrosis factor-alpha, markers of cell adhesion, dysregulation of fibrinolytic factors and the inability to regenerate from endothelial progenitor cells concomitant with increased endothelial apoptosis, increased cellular permeability and increased vascular tone. PMID- 19128226 TI - Abnormal insulin signaling: early detection of silent coronary artery disease erectile dysfunction? AB - Coronary Artery Disease (CAD) and erectile dysfunction (ED) are cardiovascular complications frequently occurring in patients with diabetes, obesity, and dyslipidemia. All these metabolic disorders are characterized by insulin resistance, defined as decreased sensitivity and/or responsiveness to metabolic actions of insulin promoting glucose disposal. Insulin resistance is not only a hallmark of metabolic abnormalities, but also a prominent feature of haemodynamic disorders. Indeed, insulin-stimulated release of endothelial factors takes part into the physiological regulation of vascular function, and altered insulin actions may profoundly affect cardiovascular homeostasis under metabolic derangement. The signpost of impaired vascular reactivity is endothelial dysfunction, a condition in which the endothelium loses its physiological ability to produce the vasodilator nitric oxide (NO). A number of molecular, cellular, physiological, and clinical studies have indicated that insulin resistance may impair NO release and damage endothelial function through several patho physiological mechanisms reciprocally interconnected. Although considered the earliest marker of impaired vascular health, endothelial dysfunction is initially asymptomatic; additional changes in the vessel structure are usually required before vascular complications manifest. Nevertheless, endothelial dysfunction may become clinically evident when endothelial-mediated relaxation is necessary and sufficient to exert a specific effect. ED may be the first expression of endothelial dysfunction, and therefore represents a sentinel event in the clinical appearance of silent CAD. Thus, insulin resistance triggers endothelial dysfunction, and endothelial dysfunction may manifest as ED long before CAD or other vascular complications become clinically evident. This review briefly outlines the main characteristics of endothelial function and dysfunction, and describes the signaling pathways involved in cardiovascular actions of insulin under physiological and pathological conditions. Moreover, potential cellular and molecular mechanisms linking insulin resistance to early CAD-ED detection are also illustrated. PMID- 19128227 TI - The mineralocorticoid receptor in endothelial physiology and disease: novel concepts in the understanding of erectile dysfunction. AB - Aldosterone is a steroid hormone that controls blood pressure by binding to the mineralocorticoid receptor (MR), a ligand-activated transcription factor, and regulating genes that play a role in salt and water homeostasis in the kidney. Dysregulation of the mineralocorticoid system reveals its crucial role in various human diseases including hypertension, atherosclerosis, cardiac failure, mineralocorticoid resistance, and disorders of the nervous system. Recently, experimental animal models of mineralocorticoid/salt-induced hypertension and atherosclerosis have revealed an epithelial, pro-inflammatory role for MR activation. Extensive investigation has begun to elucidate the mechanisms underlying the vascular effects of MR activation which involve its direct role in cardiomyocytes, vascular smooth muscle cells, and endothelial cells. More specifically, in patients with cardiovascular risk factors and disease, including diabetes, hypertension, and/or congestive heart failure, an excess of MR activation has been shown to have a negative impact on endothelial function hence disrupting the physiological balance between vasoconstriction and vasodilation. Such a mechanism may play a role in the pathogenesis of erectile dysfunction (ED), a condition that occurs frequently in patients with increased cardiovascular risk and involves endothelial dysregulation of vascular relaxation. The aim of this review is to summarize the latest concepts in MR signaling, with particular attention to the endothelium, and to discuss the potential benefits of tissue-selective MR blockade in treating subsets of ED patients, such as those with congestive heart failure and hypertension, in which the MR system may be over activated. PMID- 19128228 TI - Drugs designed to improve endothelial function: effects on erectile dysfunction. AB - Endothelial dysfunction (EtD) has emerged as a critical master pathway in the pathogenesis of both vascular disease and erectile dysfunction (ED). Drugs that have been developed for vascular diseases and/or found to have beneficial endothelial effects may be helpful in the management of ED. In this manuscript we summarize the current state of the art with respect to endothelial active drugs and discuss the evidence supporting their use in the management of ED. Pubmed query for the terms Endothelial dysfunction, erectile dysfunction, pharmaceuticals, "endothelium", "function", "pharmaceutical", "eNOS", "erectile dysfunction" and "erectile function" was conducted. Relevant articles were reviewed and summarized. A variety of cardiovascular medications have mechanisms of action that involve the endothelium. Examples include HMG-CoA Reductase inhibitors ("statins"), Angiotensin Converting Enzyme Inhibitors (ACEI), Angiotensin Receptor blockers (ARB), Endothelin Receptor Antagonists (ERA), certain beta blockers, and some oral hypoglycemics. Some of these drugs have been found to improve penile erection, although an endothelium dependent mechanism has not been conclusively demonstrated in all studies. Drugs that improve endothelial function in the cavernous arteries and the erectile tissues of the corpora cavernosa hold great promise in treating or at least minimizing the vascular damage that contributes to ED. ACEI and ARB appear to hold great promise in this regard, while statins and oral hypoglycemics may play a potentially useful role as adjunctive therapy for ED. Improvements in endothelial function may help reverse ED in some cases, which would be a marked improvement over management with currently available "on demand" ED therapies. PMID- 19128229 TI - Endothelial effects of drugs designed to treat erectile dysfunction. AB - Erectile dysfunction (ED) and endothelial dysfunction are common in individuals with multiple cardiovascular risk factors (CRFs) and are longitudinal predictors of cardiovascular events. ED is associated with systemic endothelial cell activation/dysfunction independent from CRFs or from diffuse, unrecognized vascular damage. The pathogenesis of endothelial dysfunction and ED is intimately linked through decreased expression and activation of endothelial nitric oxide (NO) synthase and the subsequent physiologic actions of NO. Furthermore, reduced biologic activity of endothelium-derived NO links atherosclerosis to ED and underscores the role of altered endothelium in the pathogenesis of both conditions. Evidence-based data suggest that daily use of phosphodiesterase type 5 inhibitors (PDE5-i) improves endothelial and erectile functions and that this benefit is lost upon drug withdrawal. Daily PDE5-i may also improve lower tract urinary symptoms related to benign prostatic hyperplasia through a reduction of adrenergic overtone. The relevance for these drugs in the prevention of complications in internal medicine diseases, i.e. cardiovascular disease, clotting disorders and autoimmune disease is uncertain. Finally, endothelial dysfunction is present in testosterone deficiency syndromes and replacement therapy is able to revert ED and to improve endothelial function. Aim of the present review is to discuss the systemic effects of drugs designed to treat ED, such as testosterone and PDE5-i, with regard to safety, unwanted effects and efficacy in improving endothelial function; finally, a goal-oriented approach to rehabilitation using daily vs. on-demand PDE5-i in difficult patients is discussed. PMID- 19128230 TI - Angiogenesis in tumor growth and metastasis. PMID- 19128231 TI - The circulating endothelial cell in cancer: towards marker and target identification. AB - Circulating endothelial cell (CEC) and progenitor (CEP) number and viability are modulated in various pathological conditions including cancer. There is increasing evidence showing that CEC and CEP play a role in cancer progression and metastasis in different animal models. At the clinical level, emerging data support that CEC and CEP kinetics and viability might predict the efficacy on anticancer drug combinations that include antiangiogenic agents. On the basis of these observations, CEC and CEP measurements have attractive potential diagnostic and therapeutic applications for malignant diseases. PMID- 19128232 TI - Tumor growth-promoting properties of macrophage migration inhibitory factor. AB - Macrophage migration inhibitor factor (MIF) is a highly conserved and evolutionarily ancient mediator with pleiotropic effects that has been implicated in tumor growth and progression. MIF's function is unique among cytokines and its effects extend to multiple processes fundamental to tumorigenesis such as tumor proliferation, evasion of apoptosis, angiogenesis and invasion. These pleiotropic functional aspects are paralleled by MIF's unique signaling properties, which involve activation of the ERK-1/2 and AKT pathways and the regulation of JAB1, p53, SCF ubiquitin ligases and HIF-1. These properties reflect features central to growth regulation, apoptosis and cell cycle control than is typical for an immune cytokine. The significance of these pro-tumorigenic properties has found support in several in vitro and in vivo models of cancer and in the positive association between MIF production and tumor aggressiveness and metastatic potential in a variety of human tumors. PMID- 19128233 TI - Pigment epithelium-derived factor prevents melanoma growth via angiogenesis inhibition. AB - Pigment epithelium-derived factor (PEDF) has recently been shown to be the most potent inhibitor of angiogenesis in the mammalian eye, and is involved in the pathogenesis of angiogenic eye disease such as proliferative diabetic retinopathy. However, a functional role for PEDF in tumor growth and angiogenesis remains to be determined. Melanoma is one of the most highly invasive and metastatic tumors. Malignant Melanoma is an increasingly common malignancy and also one the most invasive and metastatic tumors, and its mortality rates have been rapidly increasing above those of any other cancer in recent years. Surgical resection and systemic chemotherapy are the main therapeutic strategies for the treatment of malignant melanoma. However, these approaches are insufficiently effective and may be associated with significant adverse effects. Angiogenesis, a process by which new vascular networks are formed from pre-existing capillaries, is required for tumors to grow, invade and metastasize. Tumor vessels are genetically stable, and less likely to accumulate mutations that allow them to develop drug resistance in a rapid manner. Therefore, targeting vasculatures that support tumor growth, rather than cancer cells, is currently considered the most promising approach to malignant melanoma therapy. Now, novel anti-angiogenic agents with tolerable side effects are actually desired for the treatment of patients with malignant melanoma. In this paper, we review the current understanding of anti-angiogenic therapy for malignant melanoma, especially focusing on PEDF, which was recently identified as the most potent endogenous inhibitor of angiogenesis in the mammalian eye. PMID- 19128234 TI - Color-coded fluorescent protein imaging of angiogenesis: the AngioMouse models. AB - We have utilized multicolored fluorescent proteins to develop three imaging models of tumor angiogenesis. In one model, the nonluminous induced capillaries are clearly visible by contrast against the very bright tumor green fluorescent protein (GFP) fluorescence examined either intravitally or by whole-body imaging in real time. Intravital images of an orthotopic model of human pancreatic tumors expressing GFP visualized angiogenic capillaries at both primary and metastatic sites. Whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period in an orthotopic model of human breast cancer expressing GFP. Opening a reversible skin-flap in the light path markedly reduces signal attenuation, increasing detection sensitivity many-fold and enables vessels to be externally visualized in GFP-expressing tumors growing on internal organs. In another model, dual-color fluorescence imaging was effected by using red fluorescent protein (RFP)-expressing tumors growing in GFP-expressing transgenic mice that express GFP in all cells. This dual-color model visualizes with great clarity the details of the tumor-stroma interaction, especially tumor induced angiogenesis. The GFP-expressing tumor vasculature, both nascent and mature, are readily distinguished interacting with the RFP-expressing tumor cells. The third model involves a transgenic mouse in which the regulatory elements of the stem cell marker nestin drive GFP (ND-GFP). The ND-GFP mouse expresses GFP in nascent blood vessels. RFP-expressing tumors transplanted to nestin-GFP mice enable specific visualization of nascent vessels. The ND-GFP mouse was utilized to develop a rapid in vivo/ex vivo fluorescent angiogenesis assay by implanting Gelfoam which was vascularized by fluorescent nascent blood vessels. This process could be markedly stimulated or inhibited by specific compounds. We also observed, using ND-GFP mice, that the hair follicle is angiogenic and that the hair-follicle vascular network is a prime target for chemotherapy drugs which cause hair loss (chemotherapy-induced alopecia). These fluorescent models, generally termed AngioMouse, can quantitatively determine efficacy of antiangiogenesis compounds. PMID- 19128235 TI - Clinical approaches toward tumor angiogenesis: past, present and future. AB - Angiogenesis is a complex process which is critical for the growth, invasion, and metastasis of tumors. In the past ten years numerous new agents have been developed as angiogenesis inhibitors. Angiogenesis inhibitors can be classified by their targeted area of the angiogenic process; (1) VEGF and its receptors VEGFR (e.g. Bevacizumab); (2) tyrosine kinases within endothelial cells (Sunitinib); (3) proliferation of endothelial cells (Endostatin); (4) MMPs (Marimastat); (5) intercellular interactions via integrins (Cilengitide) and (6) combinations of the above mechanisms (Thalidomide). Some showed anti-tumor effects with objective responses and stable disease, and some disappeared from clinical use due to unexpected side effects or insufficient efficacies. Further investigations of combined therapies including angiogenesis inhibitors will shed light on the treatment of advanced and metastasized malignancies. PMID- 19128236 TI - Tracking stem cell therapy in the myocardium: applications of positron emission tomography. AB - The introduction of stem cells and/or progenitor cells into damaged myocardium has promising therapeutic potential in ischemic heart diseases and dilated cardiomyopathy. However, understanding the biologic mechanisms and the outcomes of transplanted cells during cardiac regenerative therapy remains mostly limited to histological assessment. Positron emission tomography (PET) is a sensitive molecular imaging modality that can non-invasively assess stem cell retention, survival, and function after transplantation. Two radiolabel approaches have been explored to implement PET: 1) direct cell labeling with a radionuclide; and 2) reporter gene-based cell labeling. Direct cell labeling has previously been used for early tracking of transplanted stem cells into the myocardium in several therapeutic clinical trials. Stem cells can also be labeled after transfection with a reporter gene, which can subsequently be visualized by using a PET reporter probe that binds to the reporter gene, therefore allowing serial in vivo evaluation of cell viability and proliferation in long-term follow-up studies. Recently, some studies successfully used this method to visualize implanted stem cells by PET imaging in animals. With the projected rapid growth of cell therapy for heart disease, PET is expected to play a major role in monitoring relevant changes that occur at every stage in cardiac regenerative therapy. These two cell tracking approaches used for PET imaging are reviewed here and compared against other imaging modalities. PMID- 19128237 TI - Subtype selectivity in phosphodiesterase 4 (PDE4): a bottleneck in rational drug design. AB - Subtype selectivity of phosphodiesterase 4 (PDE4) has been proposed to be the most salient feature for the development of drugs for asthma and inflammation. The present review provides an account of various strategies to overcome the side effects of the PDE4 inhibitors. Subtype selectivity and recent developments of molecular modeling approaches towards PDE4 were addressed using QSAR and docking, followed by a detailed structural analysis of more than three dozen available X ray structures of PDE4B and PDE4D. Usually, the lack of a 3-dimensional structure of a target protein is a bottleneck for rational drug design approaches. However, in this case the availability of 39 X-ray structures along with co-crystals has not improved the therapeutic ratio of drugs through rational drug design approaches. The investigation of structures led to find significant variations in the M-loop region, which is the integral part of the active site of PDE4B and PDE4D. These differences can be accounted for by varying conformation of the Pro(430) residue and a Thr(436)/Asn(362) mutation in the M-loop that causes variations in adjacent residue properties and also the pattern of hydrogen bonding interactions. The impact of the M-loop region on inhibitor binding has been further scrutinized by MOLCAD surfaces and hydrophobicity. These have shown that PDE4B is more hydrophobic in nature than PDE4D in the M-loop region. A review of the above aspects given the emphasis on a new PDE4 inhibitor which can access both metal and solvent pockets may possibly lead to ligands with enhanced potency. The lining of the Q2 pocket that involves the M-loop region may be considered for the design of potent subtype-selective inhibitors. PMID- 19128238 TI - Potentials of ES cell therapy in neurodegenerative diseases. AB - Neurodegenerative diseases comprise a heterogeneous spectrum of neural disorders that cause severe and progressive cognitive and motor deficits. A histological hallmark of these disorders is the occurrence of disease-specific cell death in specific regional subpopulations of neurons, such as the loss of dopaminergic neurons in the substantia nigra in Parkinson's disease. Neurodegenerative disease can also possibly occur from the loss or dysfunction of selected glial cell subsets, such as the dysfunction of supportive glial cells around somatic motor neurons in amyotrophic lateral sclerosis. The central nervous system (CNS), unlike many other tissues, has a very limited capacity for self-repair. Mature nerve cells lack the ability to regenerate, although endogenous neural stem cells exist in the adult brain that do have very limited ability to generate new functional neurons in response to injury. Rapid advances in stem cell biology have opened an alternative, fascinating perspective of neurogenesis by activation of endogenous neural stem cells and/or transplantation of in vitro-expanded stem cells and/or their neuronal- or glial-differentiated progeny. Embryonic stem (ES) cells, because of their ability to provide seemingly unlimited supply of specific cell types, their amenability to genetic engineering manipulations, and their broad developmental potential, are expected to become a cell source and biological delivery system for use in a variety of neurodegenerative diseases, and are likely to play a role in the development of novel cell-based therapies for these indications. However, before the full potential of ES cells can be realized for regenerative medicine, we need to understand mechanisms regulating their proliferation, differentiation into therapeutically relevant cells, and most importantly in the case of neuronal and glial lineages, to characterize their functional properties. In the present review we will be focusing on the factors and methodologies responsible for differentiation of ES cell into different neural precursors and neural cell lineages with particular emphasis on the potential research and clinical applications of ES cells in the field of neurodegenerative disease. PMID- 19128239 TI - Peroxiredoxin 5 confers protection against oxidative stress and apoptosis and also promotes longevity in Drosophila. AB - Peroxiredoxin 5 is a distinct isoform of the peroxiredoxin gene family. The antioxidative and anti-apoptotic functions of peroxiredoxin 5 have been extensively demonstrated in cell culture experiments. In the present paper, we provide the first functional analysis of peroxiredoxin 5 in a multicellular organism, Drosophila melanogaster. Similar to its mammalian, yeast or human counterparts, dPrx5 (Drosophila peroxiredoxin 5) is expressed in several cellular compartments, including the cytosol, nucleus and the mitochondrion. Global overexpression of dPrx5 in flies increased resistance to oxidative stress and extended their life span by up to 30% under normal conditions. The dprx5(-/-) null flies were comparatively more susceptible to oxidative stress, had higher incidence of apoptosis, and a shortened life span. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) analysis revealed that the dprx5(-/-) null mutant had discernible tissue-specific apoptotic patterns, similar to those observed in control flies exposed to paraquat. In addition, apoptosis was particularly notable in oenocytes. During development the dPrx5 levels co-varied with ecdysone pulses, suggesting inter-relationship between ecdystreroids and dPrx5 expression. The importance of dPrx5 for development was further underscored by the embryonic lethal phenotype of progeny derived from the dprx5(-/-) null mutant. Results from the present study suggest that the antioxidant and anti-apoptotic activities of dPrx5 play a critical role in development and aging of the fly. PMID- 19128240 TI - Prenatal programming of renal sodium handling in the rat. AB - Prenatally programmed hypertension induced by maternal protein restriction is associated with increased expression of the renal tubular Na+/K+/2Cl- co transporter (NKCC2) and the Na+/Cl- co-transporter (NCC). This has led to the suggestion that renal Na+ retention contributes to the development of hypertension in the LP rat (offspring exposed to a maternal low-protein diet in utero). However, this hypothesis has not been tested in vivo. Renal clearance measurements in hypertensive 4-week-old male and female LP rats showed that, although the glomerular filtration rate remained unaltered, urine flow (P<0.01) and urinary Na+ excretion rates (1.6+/-0.3 and 3.0+/-0.4 mumol.min-1.100 g-1 of body weight in control male and LP male respectively; P<0.001) were increased. Na+ excretion was positively correlated with mean arterial pressure in both males (P<0.01) and females (P<0.05), but neither the slope nor the intercept differed between control and LP rats. Fractional excretion of Na+ was increased in male (1.5+/-0.2 and 3.0+/-0.5% in control and LP rats respectively; P<0.001) and female LP rats, implying reduced tubular reabsorption of Na+. Western blotting and quantitative PCR showed that NKCC2 expression was increased, whereas NCC mRNA was not up-regulated. Na+/K+ ATPase alpha1 subunit expression did not differ from controls; however, there was a significant reduction in whole kidney pump activity (23.4+/-1.8 and 17.7+/-1.2 nmol of phosphate.mug-1 of protein.h-1 in control male and male LP rats respectively; P<0.001); immunohistochemistry showed that the alpha1 subunit was virtually absent from the inner medulla. The greater Na+ excretion of LP rats can be explained, in part, by a pressure-natriuresis mechanism; however, the loss of the Na+/K+ ATPase alpha1 subunit from the inner medulla and up-regulation of NKCC2 suggests that altered renal Na+ handling is also programmed prenatally. PMID- 19128241 TI - Effects of HCV co-infection on apoptosis of CD4+ T-cells in HIV-positive patients. AB - Apoptosis importantly contributes to loss of CD4+ T-cells in HIV infection, and modification of their apoptosis may explain why HIV/HCV (hepatitis C virus)-co infected patients are more likely to die from liver-related causes, although the effects of HCV on HIV infection remain unclear. In the present study, we studied in a cross-sectional and serial analysis spontaneous ex vivo CD4+ T-cell apoptosis in HIV/HCV-co-infected and HIV-mono-infected patients before and after HAART (highly active antiretroviral therapy). Apoptosis of peripheral blood CD4+ T-cells was measured by both a PARP [poly(ADP-ribose) polymerase] and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay to detect cells with irreversible apoptosis. Although hepatitis C alone did not increase CD4+ T-cell apoptosis, HCV co-infection disproportionately increased elevated rates of apoptosis in CD4+ T-cells from untreated HIV-positive patients. Increased CD4+ T-cell apoptosis was closely correlated with HIV, but not HCV, viral loads. Under HAART, increased rates of CD4+ T-cell apoptosis rapidly decreased both in HIV-mono-infected and HIV/HCV-co-infected patients, without any significant difference in apoptosis rates between the two patient groups after 4 weeks of therapy. Nevertheless residual CD4+ T-cell apoptosis did not reach the normal levels seen in healthy controls and remained higher in HIV patients receiving protease inhibitors than in patients with other antiretroviral regimens. The results of the present study suggest that HCV co-infection sensitizes CD4+ T-cells towards apoptosis in untreated HIV-positive patients. However, this effect is rapidly lost under effective antiretroviral therapy. PMID- 19128243 TI - Guidelines for the treatment of wounds. PMID- 19128244 TI - Guidelines to aid healing of acute wounds by decreasing impediments of healing. PMID- 19128245 TI - Autologous platelet-rich fibrin matrix as cell therapy in the healing of chronic lower-extremity ulcers. AB - A novel autologous platelet-rich fibrin matrix membrane (PRFM) was assessed for the ability to facilitate healing in patients with chronic lower-extremity ulcers. Preliminary data are presented from a prospective trial (n=21). Twelve patients were identified with 17 venous leg ulcers (VLU) and nine bearing 13 nonvenous lower-extremity ulcers. Before enrollment, the patients were evaluated for vascular status and received appropriate surgical intervention to optimize arterial and venous circulatory status. None of the ulcers had responded to a variety of standard treatments from 4 months to 53 years. Initial ulcer size ranged from 0.7 to 65 cm(2) (mean, 11.2 cm(2)). Each PRFM-treated patient received up to three applications of either a 35 or 50 mm fenestrated membrane, depending on initial ulcer size. The primary endpoints were percent and rate of complete closure as measured by digital photography, computerized planimetery, and clinical examination. Patients were followed weekly for 12 weeks with a follow-up visit at 16 weeks. At each 4-week interval, the extent of healing was assessed, and those patients with >50% reduction in wound area were allowed to continue to complete closure. Patients with <50% closure received repeated applications. Complete closure was achieved in 66.7% of the VLU patients (64.7% of treated ulcers) in 7.1 weeks (median, 6 weeks) with an average of two applications per patient. Forty-four percent complete closure was seen with non VLU patients (31% of treated ulcers). From the results of this small-scale pilot study, PRFM shows significant potential for closing of chronic leg ulcers. PMID- 19128246 TI - Pilot study using doxycycline-releasing stents to ameliorate postoperative healing quality after sinus surgery. AB - Poor postoperative healing after sinus surgery is associated with high concentrations of matrix metalloproteinase-9 (MMP-9). The frontal recess is especially vulnerable to restenosis, and frontal sinus stents have been used to overcome this problem. However, the long-term success rate is still controversial and may be poor. In this perspective, we developed doxycycline (DC)-releasing stents, delivering the MMP-9 synthesis-suppressing agent locally to the frontal recess area. We evaluated postoperative MMP-9 levels, bacterial colonization, healing quality, and symptom scores in patients suffering from chronic rhinosinusitis with (CRSwNP) and without nasal polyposis (CRSsNP) (n=10) who underwent functional endoscopic sinus surgery during which the DC-releasing and placebo stents were placed. We found that MMP-9 concentrations were significantly lower at the side of the DC-releasing stent (3,414+/-582 ng/mL) compared with the contralateral placebo stent (9,172+/-2,564 ng/mL) (p<0.05) at month 3 postsurgery. DC stents adequately suppressed bacterial growth compared with placebo stents. Furthermore, the visual analog scale (VAS) for the frontal region was significantly better (mean value 75.1 vs. 52.8, p<0.001) compared with its placebo counterpart. We conclude that compared with placebo stents, DC-releasing stents significantly lowered MMP-9 concentrations and bacterial colonization locally, and improved postoperative healing quality after functional endoscopic sinus surgery, as demonstrated by visual analog scale and ostial closure. PMID- 19128247 TI - Hedgehog signaling is essential for normal wound healing. AB - The hedgehog family of morphogens (sonic [Shh], Indian, and desert hedgehog) are central regulators of embryologic growth and tissue patterning. Although recent work implicates Shh in postnatal tissue repair and development, conclusive evidence is lacking. Here, we demonstrated the importance of Shh in wound repair, by examining the effects of cyclopamine, a specific inhibitor of the Shh signaling cascade, on tissue repair. Using a murine-splinted excisional wound model, which attenuates wound contraction in this loose-skinned rodent, we established that, by all measures (wound closure, epithelialization, granulation formation, vascularity, and proliferation), wound healing was profoundly impaired when Shh signaling was disrupted. Because embryonic disruption of Shh is associated with distinct phenotypic defects, our findings invite investigation of the potential role of Shh signaling under postnatal conditions associated with disregulated wound healing. PMID- 19128248 TI - Conservative surgical debridement as a burn treatment: supporting evidence from a porcine burn model. AB - In thermal deep-dermal burns, surgical debridement is normally used in conjunction with skin grafting or skin substitutes and debridement alone as a burn treatment is not usually practiced. The current study addresses whether or not debridement alone would enhance burn wound healing on small deep-dermal partial thickness burns. This was a prospective and blinded experimental trial using a porcine deep-dermal-partial thickness burn model. Four burns, approximately 50 cm(2) in size, were created on each of eight pigs. Two burns from each pig were immediately surgically debrided and the other two were not debrided as the internal control. Hydrate gel together with paraffin gauze were used to cover the burns for four pigs and silver dressings for the other four. Clinical assessment of wound healing was conducted over a 6-week period. Skin samples were collected at the end of the experiment and histopathological evaluation was performed. The results show thinner scar formation and lower scar height in the debrided compared with nondebrided wounds in the hydrate gel/paraffin gauze groups. There were no statistically significant differences in wound healing assessment between the debrided and nondebrided wounds dressed with silver dressings. This study provides supporting evidence that immediate debridement with an appropriate dressing and without skin grafting may promote wound healing, suggesting its potential benefit for clinical patients. PMID- 19128249 TI - A poly-herbal formulation accelerates normal and impaired diabetic wound healing. AB - In the present study, a poly-herbal formulation (PHF) was prepared by combining the aqueous lyophilized leaf extracts of Hippophae rhamnoides L. and Aloe vera L. and the ethanol rhizome extract of Curcuma longa L., in an optimized ratio (1 : 7 : 1). The efficacy of PHF treatment was studied in normal and impaired diabetic rats using a full-thickness cutaneous wound model. Topical PHF treatment increased cellular proliferation and collagen synthesis at the wound site in normal rats, as evidenced by the significant increase in DNA, total protein, hydroxyproline, and hexosamine contents in comparison with a positive control treated with a povidone-iodine ointment. The histological examinations and matrix metalloproteinases expression also correlated well with the biochemical findings, confirming the efficacy of PHF in normal wounds. In the streptozotocin-induced diabetic rats, PHF treatment increased hydroxyproline and hexosamine content. A faster wound contraction was also observed in PHF-treated normal and diabetic rats. The PHF also promoted angiogenesis as evidenced by an in vitro chick chorioallantoic membrane model and in vivo up-regulated vascular endothelial growth factor expression. The results suggest that PHF possesses significant wound healing potential in both normal as well as chronic diabetic wounds. PMID- 19128250 TI - S163 is critical for FXYD5 modulation of wound healing in airway epithelial cells. AB - The FXYD family, which contains seven members, are tissue specific regulators of the Na,K-ATPase. Increased expression of FXYD5, a cancer-cell-associated membrane glycoprotein, has been associated with increased cell motility and metastatic potential. To better understand how FXYD5 may modulate cell motility, we analyzed S163, a conserved residue in all FXYD family members located in the C-terminus. Ectopic expression of human FXYD5 S163 mutants in HEK 293 cells showed that negative charge at S163 (S163D) decreased membrane localization, assessed by immunofluorescence. Coimmunoprecipitation studies revealed decreased FXYD5/Na,K ATPase interaction for S163D compared with wild-type or S163A mutants. Interestingly, FXYD5 overexpression induced expression of vimentin, a marker of epithelial-mesenchymal transition, in murine airway epithelial cells. Because Na,K-ATPase expression is decreased in some forms of cancer and is critical for establishing cell polarity and suppressing cell motility, we analyzed S163 mutants in an epithelial cell scratch-wound model as a measure of cell migration. Wild-type FXYD5 overexpression increased reepithelialization (p<0.0001), which was further increased in S163D mutants (p<0.005). However, S163A mutants inhibited epithelial cell migration compared with wild-type FXYD5 overexpression (p<0.0001). We conclude that negative charge at S163 regulates FXYD5/Na,K-ATPase interaction and that this interaction modulates cell migration across a wound in airway epithelial cells. PMID- 19128251 TI - Antibacterial effect of dressings containing multivalent silver ion carried by zirconium phosphate on experimental rat burn wounds. AB - To evaluate the antibacterial effect of multivalent silver ion carried by zirconium phosphate (ZP-Ag) both in vitro and in an experimental rat burn wound compared with that of sulfadiazine silver (SD-Ag). Firstly, the minimal inhibition concentration and the minimal bactericidal concentration of ZP-Ag to three different strains of bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli) were compared with those of SD-Ag in vitro. Following this dressings containing the same mass of ZP-Ag or SD-Ag were applied to wounds in a rat burn model and their antimicrobial activity assayed. The inflammatory response of the wounds managed with the two kinds of dressings was compared. Both the minimal inhibition concentration and minimal bactericidal concentration of ZP-Ag to S. aureus, P. aeruginosa, and E. coli were two to four times lower than those of SD-Ag. In vivo ZP-Ag had a more effective antibacterial action inhibiting the growth of the above three bacterial strains under burn scar, compared with dressings made from the same mass of SD-Ag. Moreover, the antibacterial effect was unchanged after the ZP-Ag dressings were washed 20 times, while it dramatically decreased after the dressings with SD-Ag were washed 20 times. When covered by either regular or washed ZP-Ag dressings, the wounds showed integrity with no visible inflammatory reaction. However, various extents of inflammatory reactions on or around the wounds covered with regular SD-Ag dressing were observed. This study showed that dressings with ZP-Ag had a prominent and relative long-term antibacterial effect. ZP-Ag dressings could be an effective, low-cost management option for burn wounds. PMID- 19128252 TI - In vitro multispecies Lubbock chronic wound biofilm model. AB - Multispecies biofilms are becoming increasingly recognized as the naturally occurring state in which bacteria reside. One of the primary health issues that is now recognized to be exacerbated by biofilms are chronic, nonhealing wounds such as venous leg ulcers, diabetic foot ulcers, and pressure ulcers. Arguably three of the most important species associated with multispecies biofilms that our group sees clinically are Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus. This study was conducted to address the need for a chronic pathogenic biofilm laboratory model that allows for cooperative growth of these three organisms. We have developed a novel media formulation, simple laboratory system, quantitative polymerase chain reaction for monitoring population dynamics, and methods for objectively and subjectively measuring biofilm formation. The Lubbock chronic wound pathogenic biofilm withstood treatment with a 50-fold higher concentration of bleach than that which was completely bacteriocidal for fully turbid planktonic cultures. The Lubbock chronic wound pathogenic biofilm when treated with biofilm effectors such as gallium nitrate and triclosan responded with selective inhibition of Pseudomonas aeruginosa or Staphylococcus aureus, respectively, as has been reported in the literature. The ability of this 24-hour model to react as predicted using known biofilm effectors suggests that it will lend itself to future work in the development and testing of first-generation chronic wounds pathogenic biofilm therapeutics. We have defined a realistic in vitro multispecies biofilm model simulating the functional characteristics of chronic pathogenic biofilms and developed effective tools for its characterization and analyses. PMID- 19128253 TI - Localization of small leucine-rich proteoglycans and transforming growth factor beta in human oral mucosal wound healing. AB - Wound healing in oral mucosa is fast and results in little scar formation as compared with skin. The biological mechanisms underlying this property are poorly understood but may provide valuable information about the factors that promote wound regeneration. Small leucine-rich proteoglycans (SLRPs) decorin, biglycan, fibromodulin and lumican are extracellular matrix molecules that regulate collagen fibrillogenesis, inhibit transforming growth factor-beta (TGF-beta) activity and reduce scarring. In the present study, we analyzed accumulation of SLRPs and TGF-beta during non-scarring human oral mucosal wound healing. Biopsies were collected from healthy volunteers from unwounded tissue and from standardized experimental wounds 3-60 days postwounding. Localization of SLRPs, TGF-beta1 and TGF-beta3 was analyzed by immunohistochemical staining and quantitated by image analysis. Double immunostaining was used to study localization of SLRPs or active TGF-beta in distinct cells. Decorin, biglycan, fibromodulin, and TGF-beta isoforms showed significantly increased accumulation in the wound extracellular matrix and distinct wound cells while the abundance of lumican in the extracellular matrix was strongly reduced during wound healing. Localization and abundance of fibromodulin, lumican, and TGF-beta isoforms was also spatiotemporally regulated in the wound epithelium. The findings suggest that SLRPs regulate wound reepithelialization and connective tissue regeneration during oral mucosal wound healing. PMID- 19128254 TI - Growth factors and cytokines in wound healing. AB - Wound healing is an evolutionarily conserved, complex, multicellular process that, in skin, aims at barrier restoration. This process involves the coordinated efforts of several cell types including keratinocytes, fibroblasts, endothelial cells, macrophages, and platelets. The migration, infiltration, proliferation, and differentiation of these cells will culminate in an inflammatory response, the formation of new tissue and ultimately wound closure. This complex process is executed and regulated by an equally complex signaling network involving numerous growth factors, cytokines and chemokines. Of particular importance is the epidermal growth factor (EGF) family, transforming growth factor beta (TGF-beta) family, fibroblast growth factor (FGF) family, vascular endothelial growth factor (VEGF), granulocyte macrophage colony stimulating factor (GM-CSF), platelet derived growth factor (PDGF), connective tissue growth factor (CTGF), interleukin (IL) family, and tumor necrosis factor-alpha family. Currently, patients are treated by three growth factors: PDGF-BB, bFGF, and GM-CSF. Only PDGF-BB has successfully completed randomized clinical trials in the Unites States. With gene therapy now in clinical trial and the discovery of biodegradable polymers, fibrin mesh, and human collagen serving as potential delivery systems other growth factors may soon be available to patients. This review will focus on the specific roles of these growth factors and cytokines during the wound healing process. PMID- 19128255 TI - Vascular endothelial growth factor promotes macrophage apoptosis through stimulation of tumor necrosis factor superfamily member 14 (TNFSF14/LIGHT). AB - Resolution of inflammation is critical for normal wound healing. Inflammation is prolonged and fails to resolve properly in chronic wounds. We used in vivo and in vitro approaches to show that vascular endothelial growth factor (VEGF) induces macrophage apoptosis and to delineate mechanisms involved in this process. VEGF inhibition during wound healing leads to an increased number of macrophages remaining in wounds, suggesting the involvement of VEGF in removal of these cells from the wound. If this effect has physiological relevance, it likely occurs via apoptosis. We show that VEGF increases apoptosis of macrophages in vitro using Annexin V-FITC staining and caspase activation. Microarray analysis, reverse transcription-polymerase chain reaction, and immunoblotting showed that VEGF increases the expression of tumor necrosis factor superfamily member 14 (TNFSF14/LIGHT) in macrophages. We also show that in macrophages LIGHT promotes apoptosis through the lymphotoxin beta receptor. Moreover, inhibition of LIGHT prevents VEGF-induced death, suggesting that LIGHT mediates VEGF-induced macrophage apoptosis. Taken together, our results identify a novel role for VEGF and for LIGHT in macrophage apoptosis during wound healing, an event critical in the resolution of inflammation. This finding may lead to the development of new strategies to improve resolution of inflammation in problematic wounds. PMID- 19128256 TI - Reducing wound pain in venous leg ulcers with Biatain Ibu: a randomized, controlled double-blind clinical investigation on the performance and safety. AB - Six out of 10 patients with chronic wounds suffer from persistent wound pain. A multinational and multicenter randomized double-blind clinical investigation of 122 patients compared two moist wound healing dressings: a nonadhesive foam dressing with ibuprofen (62 patients randomized to Biatain Ibu Nonadhesive Coloplast A/S) and a nonadhesive foam without ibuprofen (60 patients to Biatain Non-Adhesive-comparator). Patients were recruited from September 2005 to April 2006. The ibuprofen foam was considered successful if the pain relief on a five point Verbal Rating Scale was higher than the comparator without compromising safety including appropriate healing rate. Additional endpoints were change in persistent wound pain between dressing changes and pain at dressing change on days 1-5 (double blind) and days 43-47 (single blind). The primary response variable, persistent pain relief, was significantly higher in the ibuprofen-foam group, as compared with the comparator on day 1-5, with a quick onset of action (p<0.05). Wound pain intensity was significantly reduced with the ibuprofen foam during day 1-5 with 40% from baseline, compared with 30% with the comparator (p<0.001). At day 43-47, the patients in the ibuprofen-foam group had a significant (p<0.05) reemergence of persistent pain and pain at dressing change (p<0.05) when the active dressing was changed to the comparator. Wound healing was similar in the ibuprofen foam and comparator group. No difference in adverse events between the comparator and the ibuprofen foam with local sustained release of low-dose ibuprofen was observed in this study. It was generally found that women reported less pain intensity than men, and pain intensity decreased with increasing age. In addition, pain intensity increased with initial pain intensity and increasing wound size. This study has demonstrated that the ibuprofen-foam dressing provided pain relief and reduced pain intensity without compromising healing or other safety parameters. PMID- 19128257 TI - The optimal temperature of first aid treatment for partial thickness burn injuries. AB - Using our porcine model of deep dermal partial thickness burn injury, various cooling techniques (15 degrees C running water, 2 degrees C running water, ice) of first aid were applied for 20 minutes compared with a control (ambient temperature). The subdermal temperatures were monitored during the treatment and wounds observed and photographed weekly for 6 weeks, observing reepithelialization, wound surface area and cosmetic appearance. Tissue histology and scar tensile strength were examined 6 weeks after burn. The 2 degrees C and ice treatments decreased the subdermal temperature the fastest and lowest, however, generally the 15 and 2 degrees C treated wounds had better outcomes in terms of reepithelialization, scar histology, and scar appearance. These findings provide evidence to support the current first aid guidelines of cold tap water (approximately 15 degrees C) for 20 minutes as being beneficial in helping to heal the burn wound. Colder water at 2 degrees C is also beneficial. Ice should not be used. PMID- 19128258 TI - Basic fibroblast growth factor accelerates and improves second-degree burn wound healing. AB - Second-degree burns are sometimes a concern for shortening patient suffering time as well as the therapeutic choice. Thus, adult second-degree burn patients (average 57.8 +/- 13.9 years old), mainly with deep dermal burns, were included. Patients receiving topical basic fibroblast growth factor (bFGF) or no bFGF were compared for clinical scar extent, passive scar hardness and elasticity using a Cutometer, direct scar hardness using a durometer, and moisture analysis of the stratum corneum at 1 year after complete wound healing. There was significantly faster wound healing with bFGF, as early as 2.2 +/- 0.9 days from the burn injury, compared with non-bFGF use (12.0 +/- 2.2 vs. 15.0 +/- 2.7 days, p<0.01). Clinical evaluation of Vancouver scale scores showed significant differences between bFGF-treated and non-bFGF-treated scars (p<0.01). Both maximal scar extension and the ratio of scar retraction to maximal scar extension, elasticity, by Cutometer were significantly greater in bFGF-treated scars than non-bFGF treated scars (0.23 +/- 0.10 vs. 0.14 +/- 0.06 mm, 0.59 +/- 0.20 vs. 0.49 +/- 0.15 mm: scar extension, scar elasticity, bFGF vs. non-bFGF, p<0.01). The durometer reading was significantly lower in bFGF-treated scars than in non-bFGF treated scars (16.2 +/- 3.8 vs. 29.3 +/- 5.1, p<0.01). Transepidermal water loss, water content, and corneal thickness were significantly less in bFGF-treated than in non-bFGF-treated scars (p<0.01). PMID- 19128259 TI - Multiplexed analysis of matrix metalloproteinases in leg ulcer tissue of patients with chronic venous insufficiency before and after compression therapy. AB - Elevated matrix metalloproteinases (MMP) levels have been implicated in the pathogenesis of chronic venous insufficiency ulcers. Quantitative measurements of a broad range of MMP proteins in human tissue treated with compression bandaging have not been reported. The goal of this study was to determine the expression of a wide range of proteases in untreated venous leg ulcer tissue and the changes in these levels after 4 weeks of high-strength compression therapy. Twenty-nine limbs with new or untreated chronic venous insufficiency and leg ulceration received therapy for 4 weeks with sustained high compression bandaging. Biopsies were obtained from healthy tissue and from ulcerated tissue before and after therapy. A novel multiplexed protein assay was used to measure multiple MMPs in a single sample. MMP protein activity, TIMP protein levels, and gene expression levels were also addressed. MMP1, 2, 3, 8, 9, 12, and 13 protein levels were elevated in ulcer tissue compared with healthy tissue. MMP8 and 9 were highly expressed in ulcer tissue. MMP3, 8, and 9 significantly decreased following treatment. Reduction in the levels of MMP1, 2, and 3 was associated with significantly higher rates of ulcer healing at 4 weeks. We conclude that compression therapy results in a reduction of the pro-inflammatory environment characterizing chronic venous ulcers, and ulcer healing is associated with resolution of specific elevated levels of protease expression. PMID- 19128261 TI - Antisense inhibition of connective tissue growth factor (CTGF/CCN2) mRNA limits hypertrophic scarring without affecting wound healing in vivo. AB - Augmented expression of connective tissue growth factor (CTGF/CCN2) is observed in healing wounds and in a variety of fibrotic disorders. It appears to enhance many of the effects of transforming growth factor-beta and has been shown to have independent fibrogenic functions. Despite these observations, its importance to dermal wound healing and the transition from wound to scar remains poorly defined. In this study, we use established rabbit models to evaluate the roles of CTGF in dermal wound healing and hypertrophic scarring. We show that CTGF mRNA demonstrates persistent up-regulation in hypertrophic scars. Treatment of wounds with antisense oligonucleotides to CTGF has no measurable effect on early wound closure. However, antisense therapy significantly limits subsequent hypertrophic scarring. Inhibition of CTGF is associated with a marked reduction in the number of myofibroblasts in scars and decreased transcription of TIMP-1 and types I and III collagen. These findings confirm CTGF to be a key mediator of hypertrophic scarring in this model. Its effect on myofibroblasts in this setting suggests a mechanism whereby it plays this role. Its limited participation in early healing implies that it may be a useful and specific target for modulating hypertrophic scarring following injury. PMID- 19128260 TI - Primary dermal fibroblasts derived from sdc-1 deficient mice migrate faster and have altered alphav integrin function. AB - ABSTRACT The goal of this study is to determine whether dermal fibroblasts lacking syndecan-1 (sdc1) show differences in integrin expression and function that could contribute to the delayed skin and corneal wound healing phenotypes seen in sdc-1 null mice. Using primary dermal fibroblasts, we show that after 3 days in culture no differences in alpha-smooth muscle actin were detected but sdc 1 null cells expressed significantly more alphav and beta1 integrin than wildtype (wt) cells. Transforming growth factor beta1 (TGFbeta1) treatment at day 3 increased alphav- and beta1-integrin expression in sdc-1 null cells at day 5 whereas wt cells showed increased expression only of alphav-integrin. Using time lapse studies, we showed that the sdc-1 null fibroblasts migrate faster than wt fibroblasts, treatment with TGFbeta1 increased these migration differences, and treatment with a TGFbeta1 antagonist caused sdc-1 null fibroblasts to slow down and migrate at the same rate as untreated wt cells. Cell spreading studies on replated fibroblasts showed altered cell spreading and focal adhesion formation on vitronectin and fibronectin-coated surfaces. Additional time lapse studies with beta1- and alphav-integrin antibody antagonists, showed that wt fibroblasts expressing sdc-1 had activated integrins on their surface that impeded their migration whereas the null cells expressed alphav-containing integrins which were less adhesive and enhanced cell migration. Surface expression studies showed increased surface expression of alpha2beta1 and alpha3beta1 on the sdc-1 null fibroblasts compared with wt fibroblasts but no significant differences in surface expression of alpha5beta1, alphavbeta3, or alphavbeta5. Taken together, our data indicates that sdc-1 functions in the activation of alphav-containing integrins and support the hypothesis that impaired wound healing phenotypes seen in sdc-1 null mice could be due to integrin-mediated defects in fibroblast migration after injury. PMID- 19128262 TI - Comparison of the effects of collagenase and extract of Centella asiatica in an experimental model of wound healing: an immunohistochemical and histopathological study. AB - In this study, we compared the effects of collagenase and Centella asiatica in the rat model. Twenty-seven female rats were divided into three groups, and two full-thickness wounds were made for each animal. Collagenase ointment was applied topically to Group I and C. asiatica ointment to Group II rats. In Group III, no treatment was applied. On the third day of treatment, wounds on the left side of three animals of each group were excised. On the fifth and eighth day of the treatments, the same procedure was performed for the remaining animals. Indirect immunohistochemical examination was performed to detect transforming growth factor beta (TGF)-beta, endothelial and inducible nitric oxide synthase (eNOS and iNOS), vascular endothelial growth factor, TGF-alpha, laminin, fibronectin, collagen I, and interleukin-1beta. According to the measurements of the wound areas and wound healing periodo, collagenase was superior to the control group. Immunohistochemical examinations showed strong (+++) iNOS and TGF-beta immunoreactivities in C. asiatica group. eNOS immunoreactivity was moderate (++) in this group. For the collagenase group, iNOS, eNOS, and TGF-beta immunoreactivities were moderate (++). In the collagenase group, while TGF-beta and iNOS immunoreactivities were weaker, laminin and fibronectin reactivities were stronger than in C. asiatica and control groups. Collagenase was superior to C. asiatica according to the immunohistochemical findings. Collagenase ointment significantly improves the quality of wound healing and scar formation and is a more appropriate treatment choice than extract of C. asiatica in the early stages of the wound healing process. PMID- 19128263 TI - Exogenous metallothionein-IIA promotes accelerated healing after a burn wound. AB - Severe injury to the epidermal barrier often results in scarring and life-long functional deficits, the outcome worsening with a number of factors including time taken to heal. We have investigated the potential of exogenous metallothionein IIA (Zn(7)-MT-IIA), a naturally occurring small cysteine-rich protein, to accelerate healing of burn wounds in a mouse model. Endogenous MT I/II expression increased in basal keratinocytes concurrent with reepithelialization after a burn injury, indicating a role for MT-I/II in wound healing. In vitro assays of a human keratinocyte cell line indicated that, compared with saline controls, exogenous Zn(7)-MT-IIA significantly increased cell viability by up to 30% (p<0.05), decreased apoptosis by 13% (p<0.05) and promoted keratinocyte migration by up to 14% (p<0.05), all properties that may be desirable to promote rapid wound repair. Further in vitro assays using immortalized and primary fibroblasts indicated that Zn7-MT-IIA did not affect fibroblast motility or contraction (p>0.05). Topical administration of exogenous Zn(7)-MT-IIA (2 microg/mL) in vivo, immediately postburn accelerated healing, promoted faster reepithelialization (3 days: phosphate-buffered saline (PBS), 8.9+/-0.3 mm diameter vs. MT-I/II, 7.1+/-0.7 mm; 7 days: PBS 5.8+/-0.98 mm vs. MT I/II, 3.6+/-1.0 mm, p<0.05) and reduced epidermal thickness (MT-I/II: 45+/-4 microm vs. PBS: 101+/-19 microm, p<0.05) compared with controls. Our data suggest that exogenous Zn(7)-MT-IIA may prove a valuable therapeutic for patients with burns and other skin injuries. PMID- 19128264 TI - Anti-apoptotic role of EGF in HaCaT keratinocytes via a PPARbeta-dependent mechanism. AB - Epidermal growth factor (EGF) plays an important role in epithelial cell proliferation and apoptosis. Our recent studies found that EGF-attenuated tumor necrosis factor-alpha induced HaCaT keratinocyte apoptosis, and this effect was accompanied by up-regulation of the expression of peroxisome proliferator activated receptor beta (PPARbeta). However, little is known about whether PPARbeta is functionally involved in the inhibition of keratinocyte apoptosis by EGF. Here, we showed that EGF up-regulated the DNA-binding and transcriptional regulation activities of PPARbeta. Antisense phosphorothioate oligonucleotides against PPARbeta markedly inhibited de novo synthesis of PPARbeta and attenuated the protective effect of EGF on tumor necrosis factor-alpha-induced apoptosis. L165041, a specific PPARbeta ligand, significantly enhanced the transcriptional regulation activity of PPARbeta and increased the protective effect of EGF. These results suggest a molecular mechanism by which EGF protects HaCaT keratinocytes against apoptosis in a PPARbeta-dependent manner. PMID- 19128265 TI - Elevated expression of isopeptide bond cross-links contributes to fibrosis in scleroderma and the healing wounds of tight skin mice. AB - Scleroderma is a chronic disease characterized by excessive tissue fibrosis. Recent studies indicate that cultured dermal fibroblasts isolated from patients produce excessive amounts of collagen and other extracellular matrix components. In this study, we investigated the mechanism(s) of abnormal extracellular matrix accumulation in the scleroderma biopsies and the healing wounds of Tsk1/+ mice. Full-thickness excisional wounds were made in Tsk1/+ and wild-type mice and were subsequently harvested at days 7, 10, and 14 postinjury. The levels of pro fibrotic cytokine, transforming growth factor were elevated in the wounds of Tsk1/+ mice. Interestingly, the levels of matrix metalloproteinase were significantly reduced in the granulation tissue of Tsk1/+ mice in comparison with wild-type. Furthermore, immunohistochemical analysis of the wounds indicated that the levels of gamma-glutamyl-epsilon-lysine cross-links were elevated in the granulation tissue of Tsk1/+ mice as well as the fibrotic lesions of scleroderma specimens. Collectively, these findings indicate that elevated collagen synthesis and decreased matrix metalloproteinase levels, in combination with increased isopeptide bond cross-links, contribute to abnormal collagen synthesis and assembly in granulation tissue of Tsk1/+ mice and the fibrotic lesions of scleroderma patients. PMID- 19128266 TI - The anti-inflammatory agent Propolis improves wound healing in a rodent model of experimental diabetes. AB - Foot ulcers and poor wound healing are problematic for patients with diabetes. The beehive protectant Propolis can improve wound healing but whether it can improve healing in diabetic wounds has not been investigated. In this study, the effect of a single application of Propolis on epithelial closure, wound morphology, cellular infiltrate, and blood vessel density were investigated. Diabetes was induced in rats using streptozocin. After 6 weeks, diabetic and control animals were wounded and the wounds were treated with Propolis or saline as control. At days 6 and 12 animals were sacrificed and wounds were excised. Compared with controls, diabetes decreased epithelial closure and reepithelialization but had no effect on wound contraction. These delays were prevented by Propolis. At day 12, the impaired macrophage infiltration (C:1.49+/ 0.09 vs. D:0.25+/-0.14), persistent neutrophil infiltration (C:0.22+/-0.19 vs. D:1.33+/-0.81), and increased myeloperoxidase activity (fourfold) in diabetic wounds were prevented by Propolis. Diabetes had no effect on wound volume, vessel number, or branch points. These novel data indicate that Propolis can accelerate wound healing in diabetes. As neutrophil infiltration is normalized, its mechanism of action may be through anti-inflammatory pathways. This result and the established safety profile of Propolis provide a rationale for studying topical application of this agent in a clinical setting. PMID- 19128267 TI - Enhanced wound healing by an epigallocatechin gallate-incorporated collagen sponge in diabetic mice. AB - Epigallocatechin-3-O-gallate (EGCG), the major polyphenolic compound present in green tea, has potent anti-oxidant and free radical-scavenging activities. In this study, various concentrations (10, 100, and 1,000 ppm) of EGCG were incorporated into a collagen sponge (CS) in order to investigate its healing effects on full-thickness wounds created in type 2 diabetic mice. After 14 days, the residual wound size of the mice treated with 10 ppm EGCG-incorporated collagen sponge (E-CS) decreased significantly faster than that of the other mice. Moreover, significant increases in the degree of reepithelialization, the thickness of the granulation tissue, and the density of the capillaries were also histologically observed in the wound sites exposed to 10 ppm E-CS in comparison with the others. Furthermore, 10 ppm E-CS resulted in significant increases in the immunoreactivity of Ki-67 (reepithelialization at the wound site), CD31 (formation of blood vessels), and alpha-smooth muscle actin (the induction of myofibroblasts across the dermis). These results suggest that a CS incorporated with EGCG at low concentrations can enhance wound healing in diabetic mice by accelerating reepithelialization and angiogenesis as well as improving the cellular reorganization of granulation tissue by triggering the activity of myofibroblasts. PMID- 19128268 TI - Staging of hypertension and total cardiovascular risk assessment: related but not the same--challenge for the hypertension specialist. PMID- 19128269 TI - Serum phosphate, blood pressure, and the metabolic syndrome--20-year follow-up of middle-aged men. AB - The authors investigated the relationship between serum phosphate (S-phosphate) and the metabolic syndrome in a group of middle-aged hypertensive and normotensive men during 20-year follow-up. Fifty-six men participated. Of the original 34 normotensive men, hypertension developed in 17. In the group as a whole and in those in whom hypertension developed, there was a significant negative relationship between S-phosphate at baseline and mean blood pressure (MBP) at follow-up. A significant relationship was observed between S-phosphate at baseline and components of the metabolic syndrome in the group as a whole, in individuals with hypertension, and in individuals with the lowest S-phosphate levels at follow-up. S-phosphate at baseline predicted MBP 20 years later in a group of hypertensive and normotensive men. When grouped according to the number of components of the metabolic syndrome, individuals with the lowest serum phosphate levels had the highest number of risk factors. These findings may suggest a role of low S-phosphate in the development of hypertension and the metabolic syndrome. PMID- 19128271 TI - Distal embolic protection during renal artery stenting: impact on hypertensive patients with renal dysfunction. AB - Distal embolic protection (DEP) may prevent embolization of atherosclerotic debris during renal artery stenting. The authors retrospectively identified 48 hypertensive patients with chronic kidney disease (CKD) who underwent renal artery stenting between 2002 and 2005 and compared stenting alone (n=17) to stenting/DEP (n=31). Blood pressure (BP) and estimated glomerular filtration rate (eGFR) (mL/min/1.73m(2)) at baseline at 6 and 12 months were compared. Overall, eGFR improved by 4.7 (P=.005) at 6 months and 3.8 (P=.003) at 12 months compared with baseline. Comparing stent to stent/DEP patients, eGFR improvement did not differ at 6 months (7.6 vs 2.9; P=.15) or at 12 months (4.4 vs 3.5; P=.74). Systolic BP reduction was similar between stent and stent/DEP patients at 6 months (-9 vs -14 mm Hg; P=.59) and at 12 months (-18 vs -16 mm Hg; P=.89). Renal artery stenting improved eGFR and systolic BP in patients with hypertension and CKD; however, DEP did not enhance these effects. PMID- 19128270 TI - Management of high blood pressure in clinical practice: perceptible qualitative differences in approaches utilized by clinicians. AB - The authors recruited a group of physicians from among the investigators participating in the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) with a greater (more successful) or lesser (less successful) proportion of trial patients meeting blood pressure (BP) control goals. The authors utilized qualitative focus group methods to identify similarities and differences in practice behaviors. Successful and less successful physicians had similarities in knowledge and practice behaviors regarding awareness of treatment guidelines, approaches to diagnosis, use of pharmacologic management, and the opinion that systolic BP guidelines should consider a patient's age. However, there were discernible differences between the two physician groups in their views on doctor-patient relationships: physicians from the less successful group were more paternalistic with their patients, while physicians from the more successful group were more likely to use a patient centered clinical approach to BP awareness and management. PMID- 19128272 TI - Evaluation and treatment of resistant or difficult-to-control hypertension. AB - An observational study was conducted in 164 patients with resistant or difficult to-control hypertension. Treatment was adjusted to achieve blood pressure recommendations from the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7). Patients were mostly male (95.73%) and African American (86.59%) and had an average age of 63.68 years. Comorbidities included diabetes (42.07%) and chronic kidney disease, with an average estimated glomerular filtration rate of 83.55 mL/min/1.73 m(2). At the time of referral, average blood pressure was 160/87 mm Hg. The average number of antihypertensive medications per patient at baseline was 3.43, which increased to 4.06 and 4.18 at 3 and 6 months, respectively (P< or =.0008), with about 80% of patients receiving a diuretic prior to intervention. Blood pressure decreased to 135.55/74.55 and 137.62/74.03 mm Hg at 3 and 6 months, respectively (P<.0001). By month 6, the blood pressure goal was reached in 45.10% of patients. At months 3 and 6, 100% of patients were now receiving a diuretic. The average estimated glomerular filtration rate at month 6 was 79.36 mL/min/1.73 m(2) (P=NS). Patients referred for resistant or difficult-to-control hypertension may be controlled by making targeted adjustments to their medical regimen. PMID- 19128273 TI - Prevalence and predictors of renal artery stenosis in hypertensive patients undergoing elective coronary procedures. AB - This study explored the prevalence and predictors of renal artery stenosis in a cohort of 525 hypertensive patients referred for elective coronary procedures. Patients underwent coronary and renal arteriography. The study defined renal artery stenosis as > or =60% luminal obstruction (physiologic or hemodynamic significance was not tested). Patients were classified into groups of those with normal renal arteries, those with insignificant renal artery stenosis, and individuals with significant renal artery stenosis. The mean age was 52.6+/-8.5 years, and 403 (76.8%) were males. Significant renal artery stenosis was found in 3.6%. It correlated significantly with hypertension duration (P=.005), history of cerebrovascular stroke (P=.01), history of angioplasty to >1 coronary vessel (P=.003), and 3-vessel coronary disease (P=.0003). Multivariate regression analysis identified 2-vessel and 3-vessel coronary artery disease as independent predictors of renal artery stenosis, with odds ratios of 4.9 and 12.1, respectively. It was concluded that invasive screening for renal artery stenosis was probably warranted only in hypertensive patients with multivessel coronary disease referred for elective coronary procedures. PMID- 19128274 TI - ASH Position Paper: Home and ambulatory blood pressure monitoring. When and how to use self (home) and ambulatory blood pressure monitoring. PMID- 19128275 TI - Using diuretics in practice--one opinion. PMID- 19128276 TI - Gingival hyperplasia in a patient with hypertension. PMID- 19128277 TI - Does treating hypertension prevent dementia? PMID- 19128278 TI - Direct renin inhibition--commonly asked questions. PMID- 19128279 TI - In angiotensin-converting enzyme inhibitor-intolerant individuals, the angiotensin receptor blocker telmisartan does not reduce the incidence of major cardiovascular events in high-risk patients: lessons learned from the Telmisartan Randomized Assessment Study in ACE-Intolerant Subjects Wtih Cardiovascular Disease (TRANSCEND). PMID- 19128280 TI - Leadership message: the ASH Specialist Program 10th anniversary. PMID- 19128282 TI - Re: "Call to action on use and reimbursement for home blood pressure monitoring: executive summary". PMID- 19128283 TI - Reprise: "One size may not fit all". PMID- 19128284 TI - Who cares for whom? Re-examining the nurse - patient relationship. PMID- 19128285 TI - Healthy babies for mothers with serious mental illness: a case management framework for mental health clinicians. AB - Women with a serious mental illness (SMI), notably schizophrenia, bipolar disorder, and personality disorders are considered high risk for adverse pregnancy and birth outcomes, which in turn, are associated with poor neurodevelopment in the child. Failure to access antenatal care, poor adherence with folate supplementation, an unhealthy lifestyle, and inappropriate health decisions can contribute to poor outcomes. Many women with SMI continue contact with mental health services while pregnant. This primary prevention project aimed to develop a framework for community mental health clinicians to improve the reproductive health outcomes for women with SMI. The consultation process involved discussions with key stakeholders, an environmental scan to determine current service delivery issues, a literature review, and individual and group interviews with community mental health clinicians, consumers, general practitioners, and midwives. Three key elements underpin the framework: early detection and monitoring of pregnancy, providing reproductive choices, and implementing a 'small known team approach' in the management of the pregnant client. Specific modules within the framework focus upon establishing a professional support network, assessing the risk of pregnancy, the early detection of pregnancy, monitoring during pregnancy, preparing for birth, and planning for the postnatal period. Implementation of the framework has the potential to significantly improve obstetric and neonatal outcomes for this high risk group. PMID- 19128286 TI - 'I'm not an outsider, I'm his mother!' A phenomenological enquiry into carer experiences of exclusion from acute psychiatric settings. AB - Contemporary standards and policies advocate carer involvement in planning, implementing, and evaluating mental health services. Critics have questioned why such standards and policies fail to move from rhetoric to reality, this particularly being applicable to carer involvement within acute psychiatric settings. As there is only limited UK research on this topic, this interpretive phenomenological study was undertaken to explore the perceived level of involvement from the perspective of carers of service users who were admitted to acute inpatient settings within the previous 2 years. Interviews were conducted with four individuals who cared for a loved one with a mental illness. The interview analysis was influenced by Van Manen, whose interpretive approach seeks to generate a deeper understanding of the phenomenon under study. Four main themes emerged: powerlessness, feeling isolated, needing to be recognized and valued, and a desire for partnership. The findings reflect the views expressed by carers in other studies, identifying that while carers seek to work in partnership with health-care professionals, at a clinical level they often feel excluded. The study concludes by discussing ways of improving and promoting carer involvement and advocating a partnership in care approach within acute psychiatry. PMID- 19128287 TI - Consumer perspectives on recovery: a focus on housing following discharge from hospital. AB - Housing is a critical element in recovery from mental illness. Without suitable housing, people have little chance of maintaining other resources in their lives, such as supportive social relationships and meaningful activities. This study investigated consumers' perspectives on the recovery needs of people who are living with a mental illness, especially those who might need supported accommodation as part of their reintegration into the community. Good quality housing is a critical element in recovery for people living with a mental illness. Findings indicate that when people become unwell, they can destroy resources in their lives, such as housing and friendships. A lack of financial stability can be a problem and exacerbate other difficulties. Having a mental illness means living with loss, stigma, and loneliness, but having someone who understands contributes significantly to recovery. The literature suggests that 'recovery' can relate to the relief of symptoms or from the stigma of the illness, recovering from the effects of treatment, from the lack of opportunities, and from the destructive aspects of mental illness. Findings from this study support these aspects, but also that recovery seems to be more. After spending time with the participants, the authors concluded that recovery also means the recovery of a life that includes supportive friends, living in a community in which at least some people 'understand', and of recovering a life that includes activities that give that life meaning. Consumers can make a significant contribution to our understanding of mental illness and recovery. PMID- 19128288 TI - Skills, knowledge, and attitudes expected of a newly-graduated mental health nurse in an inpatient setting. AB - Mental health nurses (MHN) are at the forefront of care and treatment within acute inpatient settings. Research suggests that newly-graduated MHN are unsure about the skills and knowledge that they are expected to possess, and are expected to practice at a level that they are not prepared for in their undergraduate courses; however, research focusing on competencies for newly graduated MHN is scarce. This study aimed to identify competencies expected of newly-graduated MHN working in a regional mental health facility in New South Wales, Australia. Eight registered nurses (with varying levels of experience) participated in individual semistructured interviews. Four themes were identified, and termed: communication, safety, self-awareness, and treatment. Within these four themes, 14 competencies were described that detail clear aims for new graduates, provide a better understanding of mental health nursing, and promote specialized and competent practice in mental health. PMID- 19128289 TI - Effectiveness of home visits by mental health nurses for Japanese women with post partum depression. AB - Post-partum depression affects 10-13% of Japanese women, but many do not receive appropriate treatment or support. This intervention study evaluated the effectiveness of home visits by mental health nurses for Japanese women with post partum depression. Eighteen post-partum women met the inclusion criteria and were randomly allocated into the intervention (n = 9) or control (n = 9) group at 1-2 months after giving birth. The intervention group received four weekly home visits by a mental health nurse. Control group participants received usual care. Two women in the intervention group did not complete the study. Depressive symptoms and quality of life were measured at 1 and 6 weeks' postintervention. In addition, participants completed an open-ended questionnaire on satisfaction and meaning derived from the home visits. Women in the intervention group had significant amelioration of depressive symptoms over time and reported positive benefits from the home visits, but there were no statistically significant differences between groups. Significant differences (P < 0.05) were observed at times 2 and 3 between groups in terms of increased median scores of physical, environmental, and global subscales, and the total average score of the World Health Organization/quality of life assessment instrument. On the psychological subscale, significant differences (P = 0.042) were observed between groups at time 2. The qualitative analysis of comments about home visitation revealed four categories related to 'setting their mind at ease', 'clarifying thoughts', 'improving coping abilities', and 'removing feelings of withdrawal from others'. These results suggest that home visits by mental health nurses can contribute to positive mental health and social changes for women with post-partum depression. A larger trial is warranted to test this approach to care. PMID- 19128290 TI - A comparative study of antipsychotic medication taking in people with schizophrenia. AB - Medication adherence is problematic in all chronic illnesses, none more so than in individuals with schizophrenia. The purpose of this exploratory study was to examine the factors that impacted upon antipsychotic medication taking in people with schizophrenia living in regional-rural and metropolitan Victoria, Australia, and to assess if differences existed between these two groups of participants in the factors that affected medication taking. The Factors Influencing Neuroleptic Medication Taking Scale was used with a non-probability survey sample of 81 people with schizophrenia. Ethics approval was given by university and hospital ethics committees. The results showed, overall, that there were no significant differences between the sets of participants in several demographic characteristics, insight, stigma, substance abuse, types of antipsychotic medications, significant others' support, and access to case managers and general practitioners. There were statistically significant differences between the two groups concerning living circumstances, involvement in religious/spiritual activities, perceived impact of medication side-effects, and access to psychiatrists. However, there were no statistically significant relationships between these factors and medication omission. The implications of the findings for consumers, mental health nurse case managers, families, mental health service provision, and further research, are considered. PMID- 19128291 TI - Developing the concept of family involvement and the alienation questionnaire in the context of psychiatric care. AB - Research shows that family members of people with a mental illness often experience a lack of involvement in the psychiatric care of their relative. An interpretation of the findings of these studies raises the question of whether the family members' experience of not being involved can be conceptualized in terms of alienation towards mental health services from their encounter with psychiatric care. In order to explore this possibility, the Family Involvement and Alienation Questionnaire (FIAQ) was constructed, guided by relevant theoretical frameworks and empirical research. The content validity of the questionnaire was evaluated by two groups of experienced researchers who had sound knowledge of the theoretical frameworks used. Validity based on the response process was evaluated by the parents of people with mental illness. The reliability of the questionnaire was evaluated by a test-retest design with a group of 15 family members. The data were analyzed by a non-parametric statistical method. The results of the validity and reliability evaluations showed that of the 46 original items in the questionnaire, 28 would be useful in exploring the concept of family involvement and alienation in the context of psychiatric care. Further, minor modifications could make the FIAQ useful in exploring these concepts in other settings. PMID- 19128292 TI - Contemporary mental health nursing: crisis and opportunity. PMID- 19128293 TI - Former patients' experience of psychiatric care: a qualitative investigation. PMID- 19128296 TI - Nursing & Health Sciences celebrates its 10th anniversary. PMID- 19128301 TI - Bibliometrics, citation indexing, and the journals of nursing. AB - Bibliometric research has risen in popularity during recent years and an increasing number of investigations now have examined the nursing literature. Our article provides a comprehensive overview of citation-based research in the nursing profession, as well as a discussion of bibliometrics, journal impact factors, and international publishing trends. The debate on evidence-based practice and its potential influence and relevance for nursing scholars is also covered. Although journal performance indicators are, no doubt, important for the contemporary nursing academic, it is the core research skills and attributes that nursing scholars, academics, and educators will need to consider more carefully in future if the next generation of professional nurse researchers is to truly flourish. PMID- 19128302 TI - Quality of life and physical functioning in HIV-infected individuals receiving antiretroviral therapy in KwaZulu-Natal, South Africa. AB - KwaZulu-Natal province, South Africa, accounts for 28.7% of the HIV infection total and one-third of infections among youth and children in South Africa. The purpose of this study was to examine the variables of HIV/AIDS symptoms, social support, influence of comorbid medical problems, length of time adhering to antiretroviral therapy medications, quality of life, adherence to antiretroviral medications, and physical functioning in HIV-infected individuals. Based on our model, the combination of these variables was found to determine physical functioning outcomes and adherence to HIV medications. Significant relationships were observed between physical functioning and the dependent variables of length of time on medications, comorbid health problems, and social support. A linear regression model was built to determine the degree to which these variables predicted physical functioning. In total, these predictor variables explained 29% of the variance in physical functioning. These results indicate that those individuals who reported a greater length of time on medications, fewer comorbid health problems, and greater social support had better physical functioning. PMID- 19128303 TI - Comparison of the health-promoting lifestyles of nursing and non-nursing students in Istanbul, Turkey. AB - Undertaking a health-related course at university can facilitate an awareness of health-promoting lifestyles. We carried out a descriptive and cross-sectional study with 1616 university students in Istanbul, Turkey. Students from the nursing schools were compared to those from the schools of social sciences using a Turkish version of the Health-promoting Lifestyle Profile (HPLP) II after a validation study. The nursing students had more positive health-promoting lifestyles than those of the non-nursing students. Furthermore, fourth-year nursing students had higher scores in most of the subscales of the HPLP II than did the students from the lower years; conversely, the fourth-year non-nursing students had lower scores. The sociodemographic variables, self-perceived health status, relations with family and friends, and self-perceived academic performance were associated with the HPLP. The implications are discussed for the curricula and on-campus facilities that focus on health promotion activities. PMID- 19128304 TI - Development of a fall prevention program for elderly Japanese people. AB - The purpose of this study was to develop a fall prevention program adapted to the individual risks of elderly patients in a long-term care facility by increasing the caregiving skills and motivation of the staff members. The fall prevention program was created by synthesizing information based on evidence-based practice and action research methodology was chosen to complete the development of the program. There were 31 participating patients in the intervention ward and 20 patients in the control ward. The Generalized Self-efficacy Scale and the Social Support Scale were used to evaluate fall prevention skills the motivation of the staff members. The fall prevention program helped to reduce injuries from 41.9% to 9.7% among the elderly participants and to increase emotional support and self efficacy among the staff members. Empowerment was considered to be a driving force. The fall prevention program was shown to be acceptable for use among elderly individuals in a long-term care facility. PMID- 19128305 TI - Factors influencing fatigue in Chinese nurses. AB - Factors predicting fatigue in Chinese nurses were examined in a descriptive, correlational study. The participants were 581 nurses working in general hospitals in Chengdu City, China. The study instruments included the Occupational Fatigue Exhaustion Recovery Scale, the Job Content Questionnaire, the Exposure to Hazards in Hospital Work Environments Scale, the Pittsburgh Sleep Quality Index, the Job Dissatisfaction Scale, the Beck Anxiety Inventory, and the Beck Depression Inventory. The data were analyzed by using descriptive statistics, Pearson's correlation, F statistics, and multiple regression. The findings revealed that 61.7% of the variance in chronic fatigue and 54.9% of the variance in acute fatigue were explained by the independent variables. Intershift recovery was the most important variable in the explanation of acute fatigue, while acute fatigue was the most important variable in the explanation of chronic fatigue. Different intervention strategies should be implemented regarding the different influencing factors of acute and chronic fatigue. PMID- 19128306 TI - Having knowledge of metabolic syndrome: does the meaning and consequences of the risk factors influence the life situation of Swedish adults? AB - The underlying causes of metabolic syndrome (MS) are uncertain. Knowledge from those who have experience of this syndrome should provide new insight. The aim was to explore the meaning and consequences of MS. Thirteen Swedish adults with MS, aged between 33 and 82 years, were interviewed. The interviews were analyzed using constant comparative analysis, which is the basis of grounded theory. The core category for the meaning and consequences of having the risk factors of MS consisted of the recurrence of behavior. The participants attempted to balance their insight into the causes and consequences by referring to their normal life, lifestyle, and fatalistic approach to life. Attention needs to be paid to the attitudes of the individuals with MS, as well as the known risk factors and their consequences, in order to facilitate a long-term lifestyle change in these individuals. PMID- 19128307 TI - Irish nursing students' experiences of service learning. AB - Service learning is a teaching tool that facilitates students' ability to link theory to practice while simultaneously providing a needed service to the community. This paper describes Irish nursing students' experiences of a service learning placement undertaken in a developing country. The students complete 30 h of theoretical content, which includes lectures and workshops on such topics as personal safety, health, and human rights, as well as the preparation of students for the emotional impact of the experience. All the content is underpinned by a commitment to developing reciprocal relationships with the service learning communities. To explore these students' experiences, a descriptive qualitative study was conducted. The data were collected using interviews and were analyzed by thematic analysis. Four main themes were identified: developing cultural sensitivity, caring for people in different cultures, learning/knowing more, and the potential impact on nursing practice. The findings suggest that the students are more culturally aware and are becoming more responsible citizens. PMID- 19128309 TI - Why health care needs resilient practitioners. PMID- 19128310 TI - Critical care nursing: towards 2015. PMID- 19128311 TI - Re: Critical care nursing: towards 2015. Editors' note. PMID- 19128312 TI - Clinical tools for the assessment of pain in sedated critically ill adults. AB - AIM: This paper aims to review the evidence regarding pain assessment tools for sedated patients and to establish whether the use of a tool can be recommended in practice. BACKGROUND: Pain assessment is a challenging area of critical care nursing practice, particularly among sedated patients. Tools to aid in assessing pain among this patient group have been developed and tested recently. SEARCH STRATEGY: In this systematic review five papers that tested pain assessment tools for sedated patients are discussed. These papers were identified via the CINAHL and MEDLINE databases using the search terms: 'pain assessment' and 'sedated' or 'unconscious' or 'critically ill' or 'critical illness' or 'critical care'. CONCLUSIONS: The Behavioural Pain Scale (BPS) has been tested among the broadest range of patients and was found to be a reliable and valid tool in three studies. Research is needed to further demonstrate the reliability and validity of the Critical-Care Pain Observation Tool (CPOT), as the paper of Gelinas et al. did not test its internal consistency and domain structure. The CPOT also needs testing among different critical care populations. The design of Odhner et al. study did not allow adequate testing of the Non-verbal Pain Scale (NVPS). IMPLICATIONS FOR PRACTICE: The implementation of the BPS can be recommended in intensive care units and may improve the management of pain among sedated patients by providing a systematic and consistent approach to pain assessment to guide interventions. The CPOT may also prove useful in assessing pain among sedated patients, but first requires further validation. Also, further research is needed into the effects of pain assessment tools on pain management practices and patient outcomes. PMID- 19128313 TI - Using a high-flow respiratory system (Vapotherm) within a high dependency setting. AB - BACKGROUND: Vapotherm 2000i is a non-invasive high-flow respiratory support system used mainly in the treatment of type 1 respiratory failure. It uses a mixture of oxygen and air to deliver a set concentration via nasal cannula (or tracheostomy mask). The advantage of this system is the high humidity achieved using the integral heated water system. The system has been used in neonatal practice as a replacement for conventional continuous positive airway pressure (CPAP) but there is little published research within the adult setting about its use. AIM: This study discusses the indications for the use of this non-invasive high-flow respiratory support system, the system set up, the benefits and complications of the system. AUDIT DESIGN: An audit of 72 patients was undertaken within the Surgical High Dependency Unit that collected data on the types of patient receiving the therapy, its effectiveness and patient experience. AUDIT FINDINGS: The audit demonstrated reduced respiratory rate and improvement in oxygenation for treated patients. CONCLUSION: A non-invasive high-flow respiratory support system can be effective at improving oxygenation in hypoxic patients. Patients were generally satisfied with the system and the system seems suitable for use in an adult surgical high dependency setting. PMID- 19128314 TI - Critical care nurses' workload estimates for managing patients during induced hypothermia. AB - INTRODUCTION: The purpose of this study was to provide an initial foundation for exploring how induced hypothermia impacts nursing workload in an intensive care unit setting. METHODS: This descriptive study used a questionnaire to obtain input from critical care nurses. RESULTS: The results represent 107 returned surveys from 120 surveys distributed to seven different critical care units. Nurses estimate a mean time of 9.27 min (95% CI = 5.63-12.92 min) per shift for each intervention. Nurses indicate that they typically consider employing over 10 interventions to reduce temperature or induce hypothermia (95% CI = 9.67-10.81). CONCLUSIONS: Nurses are open to using a variety of different interventions to manage temperature in critically ill patients. The time required to complete any one intervention varies significantly, but the combination of interventions most certainly has a significant impact on the workload for bedside nurses. PMID- 19128315 TI - Self-extubation risk assessment tool: predictive validity in a real-life setting. AB - AIM: To evaluate the predictive validity of the self-extubation risk assessment tool (SERAT) in intensive care patients. BACKGROUND: Unplanned extubation is an important complication in intensive care units (ICUs). Physicians and nurses working in the ICU would benefit by having access to a tool that could reliably identify patients at risk for unplanned extubation. The SERAT is a risk stratification scheme developed to identify patients at risk for deliberate self extubation. DESIGN: A prospective, diagnostic study. METHODS: Over a 3-month period, 256 patients who were admitted in one of five ICUs in four hospitals in Flanders (Belgium) were studied. The Glasgow Coma Scale and the Bloomsbury Sedation Score were completed by nurses at the start of each shift, i.e. three times per day. Independent nurse researchers collected data on planned or unplanned extubation and placed the data in the SERAT classification scheme. RESULTS: Five self-extubations and three accidental extubations occurred during the 3-month study period, yielding an incidence of 4.47% that corresponded to 0.56 unplanned extubations per 100 ventilation days. Using the highest accuracy model, we determined that the SERAT had a sensitivity of 100%, specificity of 90%, negative predictive value of 100%, positive predictive value of 1.2% and accuracy of 90%. CONCLUSIONS: Although the SERAT can correctly identify patients at risk for deliberate self-extubation, its use also produces a high number of false-positive identifications. Further research is necessary to evaluate how the false-positive rate can be reduced, and subsequently, the predictive validity of the SERAT can be improved. RELEVANCE TO CLINICAL PRACTICE: Because of the high number of false positives, the use of the SERAT in clinical practice to date is not advocated. The positive predictive value has to be improved to avoid the implementation of intensive interventions in patients who are not at risk. PMID- 19128316 TI - Commentary: Bouza C et al. (2007). Unplanned extubation in orally intubated medical patients in the ICU: a prospective cohort study. PMID- 19128318 TI - Diversity of hopanoids and squalene-hopene cyclases across a tropical land-sea gradient. AB - Bacterial hopanoids are ubiquitous in Earth surface environments. They hold promise as environmental and ecological biomarkers, if the phylogeny and physiological drivers of hopanoid biosynthesis can be linked with the distribution of hopanoids observed across a breadth of samples. Here we survey the diversity of hopanoid cyclases from a land-sea gradient across the island of San Salvador, in the easternmost part of the Bahamas. The distribution of lipids was determined for the same sites, for the first time overlaying quantification of bacteriohopanepolyols with sqhC phylogeny. The results are similar to previous reports: environmental sqhCs average < 65% translated amino acid identity to their closest named relatives, and sequences from putative Proteobacteria dominate. Additionally, a new and apparently ubiquitous group of marine hopanoid producers is identified; it has no identifiable close relatives. The greatest diversity of hopanoid lipids occurs in soil, but hopanoids represent a minor fraction of total soil-derived lipids. Marine samples contain fewer identifiable hopanoids, but they are more abundant as a fraction of the total extractable lipids. In soil, the dominant compounds are 35-aminobacteriohopane-32,33,34-triol and adenosylhopane. In an upper estuarine sample, bacteriohopanetetrol and 32,35 anhydrobacteriohopanetetrol dominate; while in lower estuarine and open marine samples, the most abundant are bacteriohopanetetrol and bacteriohopaneribonolactone. Cyclitol ethers are trace components in the soil, absent in the estuary, and of moderate abundance in the open marine setting, suggesting a dominant marine source. Conversely, aminotriol and aminotetrol decrease in abundance or disappear completely from land to ocean, while 2 methyldiplopterol shows the opposite trend. Small quantities of 2 methylbacteriohopanepolyols are detectable in all samples. The overall hopanoid distributions may correlate to the major phylogenetic families of hopanoid producers or to the environments in which they are found. PMID- 19128319 TI - Linking phylogenetic identities of bacteria to starch fermentation in an in vitro model of the large intestine by RNA-based stable isotope probing. AB - Carbohydrates, including starches, are an important energy source for humans, and are known for their interactions with the microbiota in the digestive tract. Largely, those interactions are thought to promote human health. Using 16S ribosomal RNA (rRNA)-based stable isotope probing (SIP), we identified starch fermenting bacteria under human colon-like conditions. To the microbiota of the TIM-2 in vitro model of the human colon 7.4 g l(-1) of [U-(13)C]-starch was added. RNA extracted from lumen samples after 0 (control), 2, 4 and 8 h was subjected to density-gradient ultracentrifugation. Terminal-restriction fragment length polymorphism (T-RFLP) fingerprinting and phylogenetic analyses of the labelled and unlabelled 16S rRNA suggested populations related to Ruminococcus bromii, Prevotella spp. and Eubacterium rectale to be involved in starch metabolism. Additionally, 16S rRNA related to that of Bifidobacterium adolescentis was abundant in all analysed fractions. While this might be due to the enrichment of high-GC RNA in high-density fractions, it could also indicate an active role in starch fermentation. Comparison of the T-RFLP fingerprints of experiments performed with labelled and unlabelled starch revealed Ruminococcus bromii as the primary degrader in starch fermentation in the studied model, as it was found to solely predominate in the labelled fractions. LC-MS analyses of the lumen and dialysate samples showed that, for both experiments, starch fermentation primarily yielded acetate, butyrate and propionate. Integration of molecular and metabolite data suggests metabolic cross-feeding in the system, where populations related to Ruminococcus bromii are the primary starch degrader, while those related to Prevotella spp., Bifidobacterium adolescentis and Eubacterium rectale might be further involved in the trophic chain. PMID- 19128320 TI - Insights into networks of functional microbes catalysing methanization of cellulose under mesophilic conditions. AB - DNA-SIP (stable isotope probing) was conducted on anaerobic municipal solid waste samples incubated with (13)C-cellulose, (13)C-glucose and (13)C-acetate under mesophilic conditions. A total of 567 full-length bacterial and 448 1100-bp length archaeal 16S rRNA gene sequences were analysed. In the clone libraries derived from 'heavy' DNA fractions, the most abundant sequences were affiliated with the phyla Firmicutes, Bacteroidetes, the gamma-subclass of Proteobacteria and methanogenic orders Methanomicrobiales and Methanosarcinales. Sequences related to the genus Acetivibrio (phylum Firmicutes) were recovered only in the 'heavy' DNA fraction derived from the (13)C-cellulose incubation. An oligonucleotide probe (UCL284) targeting specifically Acetivibrio was designed and used for fluorescent in situ hybridization (FISH) experiments. Interestingly, hybridization of the probe was detected in microorganisms aggregated around cellulose fibres, strengthening the conclusion that these microorganisms were major cellulose degraders. Sequences related to genus Clostridium (phylum Firmicutes) and to the family Porphyromonadaceae (phylum Bacteroidetes) were retrieved in large numbers from the 'heavy' DNA library of (13)C-Glucose incubation, suggesting their involvement in saccharide fermentation. Design and hybridization of specific FISH-probes confirmed the abundant representation of Clostridium (CLO401, CLO1248) and Porphyromonadaceae (BAC1040), which were mostly observed in the planktonic phase. Surprisingly, in the (13)C-acetate experiment, the 'heavy' DNA archaeal library was dominated by sequences related to the strictly hydrogenotrophic methanogenic genus Methanoculleus. One single operational taxonomic unit containing 70 sequences, affiliated to the gamma subclass of Proteobacteria, was retrieved in the corresponding bacterial library. FISH observations with a newly designed specific probe (UGA64) confirmed the dominance of this bacterial group. Our results show that combination of DNA-SIP and FISH applied with a series of functionally connected substrates can shed light on the networks of uncultured microbes catalysing the methanization of the most abundant chemical renewable energy source on Earth. PMID- 19128321 TI - Culture-independent evidence for the persistent presence and genetic diversity of microcystin-producing Anabaena (Cyanobacteria) in the Gulf of Finland. AB - The late summer mass occurrences of cyanobacteria in the Baltic Sea are among the largest in the world. These blooms are rarely monotypic and are often composed of a diverse assemblage of cyanobacteria. The toxicity of the blooms is attributed to Nodularia spumigena through the production of the hepatotoxic nodularin. However, the microcystin hepatotoxins have also been reported from the Baltic Sea on a number of occasions. Recent evidence links microcystin production in the Gulf of Finland directly to the genus Anabaena. Here we developed a denaturing gradient gel electrophoresis (DGGE) method based on the mcyE microcystin synthetase gene and ndaF nodularin synthetase gene that allows the culture independent discrimination of microcystin- and nodularin-producing cyanobacteria directly from environmental samples. We PCR-amplified microcystin and nodularin synthetase genes from environmental samples taken from the Gulf of Finland and separated them on a denaturing gradient gel using optimized conditions. Sequence analyses demonstrate that uncultured microcystin-producing Anabaena strains are genetically more diverse than previously demonstrated from cultured strains. Furthermore, our data show that microcystin-producing Anabaena are widespread in the open Gulf of Finland. Non-parametric statistical analysis suggested that salinity plays an important role in defining the distribution of microcystin producing Anabaena. Our results indicate that microcystin-producing blooms are a persistent phenomenon in the Gulf of Finland. PMID- 19128322 TI - Effects of alien plants on insect abundance and biomass: a food-web approach. AB - The replacement of native plants by alien species is likely to affect other trophic levels, particularly phytophagous insects. Nevertheless, the effect of alien plants on insect biomass has not yet been quantified. Given their critical role in transferring energy from plants to higher trophic levels, if alien plants do affect insect biomass, this could have far-reaching consequences for community structure. We used 35 food webs to evaluate the impacts of alien plants on insect productivity in a native forest in the Azores. Our food webs quantified plants, insect herbivores, and their parasitoids, which allowed us to test the effects of alien plants on species richness and evenness, insect abundance, insect biomass, and food-web structure. Species richness of plants and insects, along with plant species evenness, declined as the level of plant invasion increased. Nevertheless, none of the 4 quantitative food-web descriptors (number of links, link density, connectance, and interaction evenness) varied significantly with plant invasion independent of the size of the food web. Overall, insect abundance was not significantly affected by alien plants, but insect biomass was significantly reduced. This effect was due to the replacement of large insects on native plants with small insects on alien plants. Furthermore, the impact of alien plants was sufficiently severe to invert the otherwise expected pattern of species-richness decline with increased elevation. We predict a decrease in insect productivity by over 67% if conservation efforts fail to halt the invasion of alien plants in the Azores. PMID- 19128323 TI - Sea-cage aquaculture, sea lice, and declines of wild fish. AB - A sea cage, sometimes referred to as a net pen, is an enclosure designed to prevent farm fish from escaping and to protect them from large predators, while allowing a free flow of water through the cage to carry away waste. Farm fish thus share water with wild fish, which enables transmission of parasites, such as sea lice, from wild to farm and farm to wild fishes. Sea lice epidemics, together with recently documented population-level declines of wild salmon in areas of sea cage farming, are a reminder that sea-cage aquaculture is fundamentally different from terrestrial animal culture. The difference is that sea cages protect farm fish from the usual pathogen-control mechanisms of nature, such as predators, but not from the pathogens themselves. A sea cage thus becomes an unintended pathogen factory. Basic physical theory explains why sea-cage aquaculture causes sea lice on sympatric wild fish to increase and why increased lice burdens cause wild fish to decline, with extirpation as a real possibility. Theory is important to this issue because slow declines of wild fish can be difficult to detect amid large fluctuations from other causes. The important theoretical concepts are equilibrium, host-density effect, reservoir-host effect, and critical stocking level of farmed fish (stocking level at which lice proliferate on farm fish even if wild fish are not present to infect them). I explored these concepts and their implications without mathematics through examples from salmon farming. I also considered whether the lice-control techniques used by sea-cage farmers (medication and shortened grow-out times) are capable of protecting wild fish. Elementary probability showed that W ~ W* - epsilonF (where W is the abundance of wild fish, W* is the prefarm abundance, F is the abundance of farm fish, and epsilon is the ratio of lice per farm fish to lice per wild fish). Declines of wild fish can be reduced by short growing cycles for farm fish, medicating farm fish, and keeping farm stocking levels low. Declines can be avoided only by ensuring that wild fish do not share water with farmed fish, either by locating sea cages very far from wild fish or through the use of closed-containment aquaculture systems. These principles are likely to govern any aquaculture system where cage-protected farm hosts and sympatric wild hosts have a common parasite with a direct life cycle. PMID- 19128324 TI - Past and present effectiveness of protected areas for conservation of naturally and anthropogenically rare plant species. AB - The Global Strategy of Plant Conservation states that at least 60% of threatened plant species should be within protected areas. This goal has been met in some regions with long traditions of plant protection. We used gap analysis to explore how particular groups of species of conservation interest, representing different types of natural or anthropogenic rarity, have been covered by protected areas on a national scale in Estonia during the last 100 years. Species-accumulation curves indicated that plant species that are naturally rare (restricted global or local distribution, always small populations, or very rare habitat requirements) needed almost twice as many protected areas to reach the 60% target as plant species that are rare owing to lack of suitable management (species depending on grassland management, moderate forest disturbances, extensive traditional agriculture, or species potentially threatened by collecting). Temporal analysis of the establishment of protected areas suggested that grouping plant species according to the predominant cause of rarity accurately reflected the history of conservation decision making. Species found in very rare habitats have previously received special conservation attention; species dependent on traditional extensive agriculture have been largely ignored until recently. Legislative initiative and new nature-protection schemes (e.g., Natura 2000, network of protected areas in the European Union) have had a positive influence on all species groups. Consequently, the species groups needing similar action for their conservation are sensitive indicators of the effectiveness of protected-area networks. Different species groups, however, may not be uniformly conserved within protected areas, and all species groups should fulfill the target of 60% coverage within protected areas. PMID- 19128325 TI - Paediatric airway management: basic aspects. AB - Paediatric airway management is a great challenge, especially for anaesthesiologists working in departments with a low number of paediatric surgical procedures. The paediatric airway is substantially different from the adult airway and obstruction leads to rapid desaturation in infants and small children. This paper aims at providing the non-paediatric anaesthesiologist with a set of safe and simple principles for basic paediatric airway management. In contrast to adults, most children with difficult airways are recognised before induction of anaesthesia but problems may arise in all children. Airway obstruction can be avoided by paying close attention to the positioning of the head of the child and by keeping the mouth of the child open during mask ventilation. The use of oral and nasopharyngeal airways, laryngeal mask airways, and cuffed endotracheal tubes is discussed with special reference to the circumstances in infants. A slightly different technique during laryngoscopy is suggested. The treatment of airway oedema and laryngospasm is described. PMID- 19128326 TI - Neurological deficit after thoracic epidural catheter insertion. PMID- 19128327 TI - Combined sciatic, femoral and obturator nerve blocks for an infra-inguinal arterial bypass graft surgery. PMID- 19128328 TI - A case of anesthesia mumps after general anesthesia. PMID- 19128329 TI - Combitube and Easytube should be included in the Scandinavian guidelines for pre hospital airway management. PMID- 19128330 TI - The accomplishments of 2008 and the opportunities and challenges for 2009. PMID- 19128331 TI - Coining and defining novel nursing terminology. Part 2: critical incident nursing intervention. AB - PURPOSE: In the second of a three-part series, a novel nursing terminology is introduced and proposed for inclusion in the Nursing Interventions Classification (NIC): Critical incident nursing intervention (CINI), defined as any indirect or direct care registered nurse-initiated treatment, based upon clinical judgment and knowledge that a registered nurse performs in response to a critical incident nursing diagnosis (CIND). A CIND is defined as recognition of an acute life threatening event that occurs as a result of disease, surgery, treatment, or medication. DATA SOURCES: The literature, research studies, meta-analyses from a variety of disciplines, and personal clinical experience serve as the data sources for this article. DATA SYNTHESIS: The current nursing interventions in the NIC are inaccurate or inadequate for describing nursing care during life threatening situations. The lack of standardized nursing terminology creates a barrier that may impede critical communication and patient care during life threatening situations. CONCLUSION: Coining and defining novel nursing terminology, CINI, for patient care during life-threatening situations is important and fills the gap in the current standardized nursing terminology. IMPLICATIONS FOR NURSING PRACTICE: Refining the NIC will permit nursing researchers, among others, to conduct studies on nursing interventions in conjunction with the proposed novel nursing terminology, CINI. The first article in this series (Part 1) introduced the novel nursing terminology: CIND; the present article (Part 2) introduces the novel nursing terminology: CINI; and the third article in this series (Part 3) will introduce the novel nursing terminology: critical incident control. PMID- 19128332 TI - Diagnoses and interventions pertinent to intellectual disability nursing. AB - PURPOSE: The field of intellectual disability care in Ireland has been undergoing significant change, and this has called into question the role of specialist intellectual disability nursing. This review aims to identify the diagnoses and interventions that are employed by intellectual disability nurses. DATA SOURCES: Exploration of the relevant professional literature, drawing on a broad scope of sources, was completed. DATA SYNTHESIS: Examination and discussion within the taxonomic structure of the Nursing Interventions Classification. CONCLUSIONS: Initial data set of diagnoses and interventions identified as a basis and justification for further systematic examination of specialist intellectual disability nursing practice. IMPLICATIONS FOR NURSING PRACTICE: Explication and elaboration of the contribution of specialist nursing within intellectual disability care settings. PMID- 19128333 TI - Factors that inhibit the use of nursing language. AB - PURPOSE: The purpose of this study was to identify the impact of institutional, professional, and personal factors on nurses and on their efforts to make nursing diagnoses. METHODS: A cross-sectional study was conducted in which all participants completed a questionnaire established to measure each individual group of factors. A score (0-100) was established to measure each group of factors. RESULTS: Twenty-one nurses responded that their busy shifts, the number of patients per nurse, and their involvement with administrative tasks were the three parameters most frequently identified as interfering with implementing nursing diagnosis. CONCLUSION: The recognition of these factors and improved institutional support may facilitate the implementation of nursing diagnoses. PRACTICE IMPLICATIONS: Electronic records may contribute to the implementation of the nursing process, particularly if the identified signs and symptoms are documented and then linked to nursing diagnoses and interventions. PMID- 19128334 TI - Current knowledge related to intelligence and thinking with implications for the development and use of case studies. AB - PURPOSE: This paper reviews current knowledge regarding intelligence and thinking, and relates this knowledge to learning to diagnose human responses and to select health outcomes and nursing interventions. DATA SOURCES: Knowledge from relevant literature sources was summarized. DATA SYNTHESIS: The provision of high quality nursing care requires use of critical thinking with three elements of nursing care: nursing diagnosis, health outcomes, and nursing interventions. Metacognition (thinking about thinking) should be used with knowledge of the subject matter and repeated practice in using the knowledge. Because there are limited clinical opportunities to practice using metacognition and knowledge of these nursing care elements, case studies can be used to foster nurses' expertise. CONCLUSIONS: Simulations of clinical cases are needed that illustrate application of the nursing knowledge represented in NANDA International, Nursing Outcomes Classification, and Nursing Interventions Classification. IMPLICATIONS: The International Journal of Nursing Terminologies and Classifications will promote the dispersion of case studies as a means of facilitating the implementation and use of nursing languages and classifications. PMID- 19128335 TI - Using standardized nursing languages: a case study exemplar on management of diabetes mellitus. AB - PURPOSE: This case study illustrates the characteristics of the nursing diagnosis readiness for enhanced self health management. DATA SOURCES: Published literature, experience, and expertise of the author were utilized as data sources. DATA SYNTHESIS: The standardized nursing languages of NANDA International, Nursing Outcomes Classification, Nursing Interventions Classification, and interventions related to the client's management of diabetes were derived for the case study. CONCLUSION: The use of standardized nursing languages with interventions related to the management of type 2 diabetes provided a framework for nurses to support clients with enhanced self-management to improve their outcomes. IMPLICATIONS FOR NURSING PRACTICE: The use of standardized nursing language in the management of type 2 diabetes can improve client outcomes. PMID- 19128336 TI - The importance of managing cardiovascular risk in the treatment of hypertension: the role of ACE inhibitors and ARBs. PMID- 19128337 TI - Blood cultures for febrile patients in the acute care setting: too quick on the draw? AB - PURPOSE: To review the fever literature and determine how 38.3 degrees C was deemed the optimal fever threshold that predicts bacteremia. DATA SOURCES: PubMed, MEDLINE, Cochrane database, and the Cumulative Index to Nursing and Allied Health. CONCLUSIONS: A temperature of 38.3 degrees C has come to be the threshold value that typically triggers diagnostic fever evaluation for bacteremia in hospitalized patients. Studies that define predictors of bacteremia provide conflicting results, and most bacteremia predictor models have not been externally validated. Therefore, current fever guidelines are based on consensus opinion rather than large clinical trials identifying a specific threshold with high sensitivity and a high negative predictive value. IMPLICATIONS FOR PRACTICE: The use of a single temperature threshold of 38.3 degrees C for the prediction of bacteremia is not sufficient in all patients. Additional factors should be considered, including patient population, supporting clinical signs and symptoms, and the patient's medical history. PMID- 19128338 TI - Postpartum management for gestational diabetes mellitus: policy and practice implications. AB - PURPOSE: The pathogenesis of gestational diabetes mellitus (GDM) and insulin resistance, risk factors, links to diabetes following the pregnancy, and clinical practice recommendations for this population are reviewed. Economic implications and future research needs are included. DATA SOURCES: Review of original studies and meta-analyses from Medline, PubMed, CINAHL, Cochrane Library Online, and other scientific and public health literature; personal clinical experience with the population and dissertation research with women who experience GDM. CONCLUSIONS: GDM is a common condition experienced during pregnancy that is associated with long-term maternal risks for the development of type 2 diabetes following the index pregnancy. For the woman with GDM, the trajectory of diabetes development can often be prevented or postponed with comprehensive education and lifestyle changes, but there is evidence to suggest that preventive efforts directed at this population can be improved. Because the prevalence of type 2 diabetes and glucose intolerance as well as the costs of treatment of these disorders are escalating in the United States, attention should be focused on this vulnerable population. IMPLICATIONS: Nurse practitioners need to be cognizant of the risks for diabetes and should follow clinical practice recommendations for routine screening and implementing preventive strategies for women who experience GDM. Office reminder systems such as standardized documentation flow sheets and electronic medical records may assist with provider adherence to practice recommendations. Strict postpartum screening and management may help reduce risks for developing diabetes in this vulnerable population. PMID- 19128339 TI - Barriers to health care and health-seeking behaviors faced by Black men. AB - PURPOSE: To describe unique barriers to health care and health-seeking behaviors faced by Black men and to make recommendations to healthcare providers for improving healthcare access for Black men. DATA SOURCES: A PubMed search for research articles published after 1999 was conducted. CONCLUSIONS: The articles selected for this state of the science clinical paper addressed barriers to access faced by Black men and/or management strategies used to mitigate barriers and improve access. The barriers identified include socioeconomic status, masculinity, racism, lack of awareness of the need for primary care, religious beliefs, and peer influences. As singular entities, these may not appear unique, but when viewed collectively, they represent an overwhelming constellation of obstacles for Black men. IMPLICATIONS FOR PRACTICE: Individual, community, state, and national level recommendations for nurse practitioner actions to improve healthcare access for Black men include public service announcements, radio commercials, and billboards aimed at raising awareness of healthcare issues in the Black community, provision of preventive services through health fairs, and development of positive provider-patient relationships. PMID- 19128340 TI - An unusual cause of knee pain discovered at a nurse practitioner clinic. AB - PURPOSE: To describe an interesting and instructive case of knee pain from nurse practitioner (NP) practice and discuss the epidemiology, pathophysiology, clinical evaluation, and treatment of osteosarcoma. DATA SOURCES: Findings from the history, physical examination, diagnostic testing, and follow-up of the case of an 18-year-old male who first presented with nontraumatic, diffuse left knee pain of 2.5-month duration. CONCLUSIONS: At follow-up, after a trial of conservative treatment, a lesion suspicious for osteosarcoma was seen in the proximal tibia. Osteosarcoma is a rare but dangerous cause of chronic extremity pain, especially in children and adolescents. IMPLICATIONS FOR PRACTICE: NPs must consider malignant bone tumors in the differential diagnosis of traumatic and nontraumatic extremity pain of extended duration, especially in children and adolescents. Thorough, persistent follow-up on recommended tests and referrals is necessary to ensure that important findings are not missed. The care of uninsured patients requires particular attention to cost concerns and access issues. PMID- 19128341 TI - Facilitating risk reduction among homeless and street-involved youth. AB - PURPOSE: The purposes of this evaluation project were to describe a group of homeless adolescents and street-involved youth who utilized a mobile unit that provided medical and mental healthcare services and to assess the efficacy of the services provided in reducing their health risk behaviors. DATA SOURCES: The records of 95 youth aged 15-25 years who used the medical mobile unit for an average of 14 months were examined and evaluated according to the national health indicators related to risk reduction. Current literature related to health risk behavior among homeless youth was reviewed, synthesized, and provided the background for this article. CONCLUSIONS: Data were obtained from the records of mostly heterosexual youth with a mean age of 20.5 years. Approximately one third of the participants were high school graduates and most were without health insurance. Living situations were transient including friends, shelters, crash pads, or the streets. Abuse accounted for the majority leaving home. Psychiatric conditions and substance abuse were common. Medical conditions were related to transient living situations, substance abuse, and sexual activity. Success of the program was associated with sustained counseling, stabilizing youth on psychotropic medications, decreasing substance use, providing birth control and immunizations, and treating medical conditions. IMPLICATIONS FOR PRACTICE: Homeless youth are one of the most underserved vulnerable populations in the United States with limited access and utilization of appropriate healthcare services. Nurse practitioners often serve as care providers but are also in a position to effectively lobby to improve health care for homeless youth through professional organizations and community activism. Furthermore, when designing and evaluating healthcare services, multidisciplinary teams need to consider risk reduction for homeless youth in the context of their environment. PMID- 19128343 TI - Second language learning in a family nurse practitioner and nurse midwifery diversity education project. AB - PURPOSE: To describe the Clinical Communication Program developed to integrate second language learning (L2), multimedia, Web-based technologies, and the Internet in an advanced practice nursing education program. DATA SOURCES: Electronic recording devices as well as audio, video editing, Web design, and programming software were used as tools for developing L2 scenarios for practice in clinical settings. CONCLUSIONS: The Clinical Communication Program offers opportunities to support both students and faculty members to develop their linguistic and cultural competence skills to serve better their patients, in general, and their students who speak a language other than English, in particular. The program provided 24 h on-demand access for using audio, video, and text exercises via the Internet. IMPLICATIONS FOR PRACTICE: L2 education for healthcare providers includes linguistic (listening, speaking, reading, and writing) experiences as well as cultural competence and practices inside and outside the classroom environment as well as online and offline the Internet realm. PMID- 19128344 TI - Improving patient outlook in rheumatoid arthritis: experience with abatacept. AB - PURPOSE: To examine the importance of improving patient outlook in rheumatoid arthritis (RA) and to discuss the role of the nurse practitioner (NP) who, through the assessment of patient-reported outcomes and in acting as an advocate for the patient with the wider healthcare team, has a crucial part to play in managing the overall well-being of the patient. This article will draw on the clinical experience to date with abatacept, a first-in-class therapy that has been approved for the treatment of RA in patients with an inadequate response to either traditional disease-modifying antirheumatic drugs (DMARDs), such as methotrexate, or biological DMARDs, such as tumor necrosis factor-alpha antagonists. DATA SOURCES: A comprehensive literature search was performed using the National Library of Medicine (MEDLINE), EMBASE, and BIOSIS databases (restricted to articles posted between January 2000 and February 2007) with the search terms CTLA-4Ig, abatacept, and primary clinical trial publications in patients with RA. The clinical data are summarized in this review along with safety data presented in the prescribing information. CONCLUSIONS: Recent changes in the approach to RA treatment, particularly the advent of biological therapies, have impacted the role of the NP. The role of the NP is integral to the management of RA and in maximizing patient outcomes, through educating patients to make informed choices regarding their treatment, ensuring the safe administration of therapies and monitoring response to therapy, and in acting as an advocate for the patient within the wider healthcare team. IMPLICATIONS FOR PRACTICE: The use of more patient-centered measures of response are gaining increasing importance both in clinical trials and in clinical practice, and as such the NP has an important role in ensuring that both the physical and the psychological needs of patients are met. Clinical trials to date have shown that abatacept provides significant and clinically meaningful improvements in patient reported outcomes, as well as demonstrating significant clinical benefits and a consistent safety profile, thus representing a valuable treatment option within the RA treatment armamentarium. PMID- 19128345 TI - Clinical management of HIV/hepatitis C virus coinfection. AB - PURPOSE: The purpose of this study was to review the current management of hepatitis C virus (HCV) in persons coinfected with HIV. DATA SOURCES: Comprehensive review of current scientific literature derived from electronic databases, article bibliographies, and conference abstracts. CONCLUSIONS: HCV treatment is feasible in the individual coinfected with HIV; however, therapy is complex and requires intensive monitoring and support to achieve the outcome of viral eradication. New strategies to improve HCV treatment rates, adherence to therapy, and virological response rates are needed in this patient population. IMPLICATIONS FOR PRACTICE: Nurse practitioners are crucial to the management of the HIV/HCV-coinfected patient. This patient population requires detailed clinical monitoring, education, side effect management, and strategies to improve adherence to therapy. PMID- 19128346 TI - Diastolic heart failure: state of the science on best treatment practices. AB - PURPOSE: The purpose of this article is to increase awareness among nurse practitioners (NPs) of the current state of the science on diastolic heart failure (DHF), the American College of Cardiology (ACC) and the American Heart Association (AHA) guidelines for DHF, and pathophysiology, diagnosis, and nonpharmacological and pharmacological management of DHF. DATA SOURCES: The articles included in the review of the state of the science were retrieved by a search of PUBMED literature using the following key search terms: heart failure, diastolic heart failure, preserved systolic function, heart failure management, treatment of diastolic heart failure, treatment of diastolic dysfunction, and treatment of preserved systolic function. Current published guidelines from the ACC and AHA were reviewed to establish clinical recommendations for patients with DHF. CONCLUSIONS: The state of the science and clinical recommendations for DHF are in the early stages compared to those for systolic heart failure (SHF). The need for more randomized clinical trials on nonpharmacological and pharmacological management and the development of standardized guidelines for DHF patients are clearly apparent. IMPLICATIONS FOR PRACTICE: Both nonpharmacologic and pharmacologic management are effective and necessary to control the clinical signs and symptoms of DHF and improve overall quality of life. Successful tailoring of a treatment plan to suit each individual patient's needs and including the family are important for the NP to consider. PMID- 19128347 TI - Why we do what we do: implementation of practice guidelines by family nurse practitioner students. AB - PURPOSE: To examine who or what influenced family nurse practitioner (FNP) students' implementation of Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of Blood Pressure (JNC-7) guidelines for hypertension management. DATA SOURCES: Eleven FNP students in their final semester completed a clinical course assignment in which they each provided care to five patients with hypertension (N = 55). Written responses to a 10-item tool eliciting patient management data as well as perceived barriers or facilitators to using JNC-7 clinical guidelines in precepted clinical settings were analyzed. CONCLUSIONS: The extent to which JNC-7 guidelines predicted clinical management of hypertension by student FNPs in primary care settings was congruent with the literature findings, reflecting a wide variation in implementation of guidelines from evidence-based practice (EBP) data. The literature supports utilization of clinical guidelines in patient care. Yet, implementation of guidelines in clinical practice is a complex phenomenon. IMPLICATIONS FOR PRACTICE: Practitioners are looking for reliable evidence on which to base decision making. In the long term, EBP addresses some of the complexities of effective care delivery. The current trend is toward increasing utilization of evidence in caring for our patients. Implementation of EBP guidelines by preceptors in the workplace should increase as NP preceptors serve as role models of EBP implementation. Rewards may be important factors in implementation. PMID- 19128350 TI - Secretoglobin 1A1 gene and asthma pre-disposition: what is the evidence? PMID- 19128351 TI - Do helminth parasites protect against atopy and allergic disease? AB - Allergic diseases are rare in areas with high helminth parasite exposure and common where helminth exposure is lacking or significantly reduced, such as urban areas of developing countries and industrialized nations. Studies suggest that helminths induce a systemic immuno-modulatory network, including regulatory T cells and anti-inflammatory IL-10, which might play a key role in the protection against the allergic phenotype. Here, we review the current cross-sectional, birth cohort, and intervention study evidence for a protective effect of helminth infection on allergy. There is increasing evidence for a causal relationship between helminth infection and reduced skin prick test responsiveness to allergens. Cross-sectional studies have shown a consistent negative relationship, and these results have been confirmed in several, although not all, intervention studies. The immunological basis for this protective effect is less clear. Recent studies do not support the mast-cell IgE saturation hypothesis, but suggest that protection is associated with IL-10 production. As for allergic disease, cross sectional studies support a negative relationship between clinical asthma and infection with some helminth species, particularly hookworm, but more studies are required to draw conclusions for eczema and rhinitis. In addition, none of the few intervention studies to date have demonstrated an increase in clinical allergy after helminth treatment, and further studies are needed. Furthermore, we are only beginning to understand the host genetic factors that are potentially involved. A genetically predetermined T-helper type 2 cell-dominated cytokine milieu reduces parasite burden and may enhance host survival in an environment where helminth parasites are prevalent. Lack of parasite exposure in such hosts might lead to hypersensitivity to seemingly minor environmental allergen stimuli. Large birth cohort studies in helminth-endemic areas that use epidemiological, genetic, and immunological tools are required to further examine how helminth parasites affect the development of atopy and allergic disease. Intervention studies with hookworm in parasite-naive allergic individuals are currently ongoing in the United Kingdom to test the above hypotheses further. PMID- 19128352 TI - BSACI guidelines for the management of drug allergy. AB - These guidelines have been prepared by the Standards of Care Committee (SOCC) of the British Society for Allergy and Clinical Immunology (BSACI) and are intended for allergists and others with a special interest in allergy. As routine or validated tests are not available for the majority of drugs, considerable experience is required for the investigation of allergic drug reactions and to undertake specific drug challenge. A missed or incorrect diagnosis of drug allergy can have serious consequences. Therefore, investigation and management of drug allergy is best carried out in specialist centres with large patient numbers and adequate competence and resources to manage complex cases. The recommendations are evidence-based but where evidence was lacking consensus was reached by the panel of specialists on the committee. The document encompasses epidemiology, risk factors, clinical patterns of drug allergy, diagnosis and treatment procedures. In order to achieve a correct diagnosis we have placed particular emphasis on obtaining an accurate clinical history and on the physical examination, as these are critical to the choice of skin tests and subsequent drug provocation. After the diagnosis of drug allergy has been established, communication of results and patient education are vital components of overall patient management. PMID- 19128353 TI - Cross-sectional and longitudinal association of the secretoglobin 1A1 gene A38G polymorphism with asthma phenotype in the Perth Infant Asthma Follow-up cohort. AB - BACKGROUND: Associations between Clara cell secretory protein gene variants (SCGB1A1, also known as CC16, CC10, CCSP and uteroglobin) and the asthma phenotype have been found in five out of eight studies world-wide. No study has investigated the contribution of SCGB1A1 polymorphisms to the development and/or persistence of the asthma phenotype in a birth cohort followed over time. OBJECTIVE: The aim of this study was to determine the role of the SCGB1A1 gene in the development of the asthma phenotype. METHODS: The Perth Infant Asthma Follow up (PIAF) cohort (n=231 unrelated infants, unselected for asthma and recruited at birth) were seen at 1 month, 6 and 11 years of age, and had a questionnaire, lung function, airway responsiveness (AR) and skin prick tests (SPTs) completed. Blood was taken at 6 and 11 years for total and specific immunoglobulin E (sIgE) and DNA extraction. SPT positivity had at least one positive SPT. SIgE>4 kU/L had at least one sIgE above 4 kU/L. SCGB1A1 A38G (rs3741240), that alters gene transcription, was genotyped using Sau96I restriction digestion of exon 1 PCR products. RESULTS: At 6 and 11 years of age, 33.0% and 29.7% of those genotyped had doctor-diagnosed asthma, and 35.8% and 52.1% had SPT positivity. In cross sectional analyses, children with 38G/38A or 38A/38A had increased AR at 1 month (1.72-fold, P=0.013); sIgE>4 kU/L [odds ratio (OR)=6.95, 95% confidence interval (CI)=1.35-35.91, P=0.021]; house dust mite (HDM) SPT positivity (OR=7.21, 95% CI=1.09-47.78, P=0.041) and sIgE (4.57-fold, P=0.045) at 6 years; and doctor diagnosed asthma (OR=3.93, 95% CI=1.24-12.47, P=0.02) and cat SPT positivity (OR=4.34, 95% CI=1.01-18.77, P=0.049) at 11 years. Longitudinal analyses of 6 and 11 years paired data showed that children with 38A/38A had increased persistent sIgE>4 kU/L (OR=11.87, 95% CI=1.97-71.53, P=0.007) and persistent HDM SPT positivity (OR=7.84, 95% CI=1.04-58.92, P=0.045). CONCLUSION: SCGB1A1 A38G may play a role in the development and persistence of the asthma phenotype in childhood. PMID- 19128354 TI - Increased production of cysteinyl leukotrienes and prostaglandin D2 during human anaphylaxis. AB - BACKGROUND: Anaphylaxis is a life-threatening syndrome resulting from the sudden release of mast cell- and basophil-derived mediators into the circulation. However, pathological evidence of the association between inflammatory mediators and human anaphylaxis is insufficient. OBJECTIVE: The aim of this study was to better understand the relationship between in vivo production of inflammatory mediators and the pathogenesis of anaphylaxis. We also sought to evaluate mast cell activation in anaphylaxis. METHODS: We measured the concentrations of various inflammatory mediators in urine samples, which were collected from 32 anaphylactic patients during the onset of anaphylaxis and during clinical remission, 21 patients with asthma on acute exacerbation and 15 healthy control subjects. Blood and urine specimens were collected from the patients after provocation test. Urinary leukotriene E4 (LTE4), 9alpha, 11beta-prostaglandin F2 (9alpha, 11beta-PGF2), eosinophil-derived neurotoxin (EDN) and leukotriene B4 glucuronide (LTBG) concentrations were determined by enzyme immunoassay, and the activity of plasma platelet-activating factor acetylhydrolase and serum tryptase concentration were measured using commercially available kits. RESULTS: Significantly higher concentrations of urinary LTE4 and 9alpha, 11beta-PGF2, which immediately decreased during clinical remission, were observed in the anaphylactic patients than in asthmatic patients on acute exacerbation and healthy control subjects. Concentrations of EDN and LTBG were not significantly different among the anaphylactic patients, asthmatic patients on acute exacerbation and healthy subjects. There was a significant correlation between urinary LTE4 and 9alpha, 11beta-PGF2 concentrations in the anaphylactic patients (r=0.672, P=0.005, n=32). In addition, LTE4 concentration in patients with anaphylactic shock is significantly elevated compared with that in patients without anaphylactic shock. CONCLUSIONS: This is a report on the significant increase in urinary LTE4 and 9alpha, 11beta-PGF2 concentrations during anaphylaxis. Urinary LTE4 and 9alpha, 11beta-PGF2 concentrations may be a reliable marker of endogenous production of inflammatory mediators associated with anaphylaxis. PMID- 19128355 TI - Soybean beta-conglycinin as the main allergen in a patient with food-dependent exercise-induced anaphylaxis by tofu: food processing alters pepsin resistance. AB - BACKGROUND: Food-dependent exercise-induced anaphylaxis (FDEIA) due to soybeans is a rare disorder. The allergen responsible for FDEIA due to soybeans has not yet been determined. OBJECTIVE: We characterized the clinical features of a patient with FDEIA due to tofu, who was well tolerant to drinking soy milk. We then sought to identify the responsible soybean allergen(s) in that patient. We further studied whether different stabilities of the allergen(s) to pepsin digestion between two soybean products are related to their clinical allergenicity. METHODS: Skin prick tests and provocation tests using soybean products were performed to detect the responsible food and other factors that induced the allergic symptoms. Specific IgE to various soybean allergens were examined by ImmunoCAP, ELISA and protein microarray assays. Immunoblotting for soybeans and soybean products using the patient's serum was also performed. Soybean products were serially digested by pepsin to disclose the stability of the allergens. RESULTS: Provocation with ingestion of tofu and exercise induced the allergic symptoms, while ingestion of soy milk and exercise did not. Immunoblot analysis, ELISA and protein microarray assay revealed that beta conglycinin mainly reacts with IgE antibodies in the patient's serum. By immunoblot analysis, beta-conglycinin in soy milk completely disappeared after pepsin digestion within 20 min, whereas beta-conglycinin in tofu was almost intact after more than 120 min of pepsin digestion. CONCLUSION: We identified beta-conglycinin as the causative allergen in a patient with FDEIA induced by tofu. The difference in resistance to pepsin digestion between tofu and soy milk suggests that the presence of undigested allergens in the digestive tract is a prerequisite for the development of FDEIA. PMID- 19128356 TI - The T-cell receptor repertoire of regulatory T cells. AB - The CD4(+) CD25(+) regulatory population of T cells (Treg cells), which expresses the forkhead family transcription factor (Foxp3), is the key component of the peripheral tolerance mechanism that protects us from a variety of autoimmune diseases. Experimental evidence shows that Treg cells recognize a wide range of antigenic specificities with increased reactivity to self antigens, although the affinity of these interactions remains to be further defined. The Treg repertoire is highly diverse with a distinct set of T-cell receptors (TCRs), and yet is overlapping to some extent with the repertoire of conventional T cells (Tconv cells). The majority of Treg cells are generated in the thymus. However, the role of the TCR specificity in directing thymic precursors to become Treg or Tconv cells remains unclear. On the one hand, the higher self reactivity of Treg cells and utilization of different TCRs in Treg and Tconv repertoires suggest that in TCR interactions an initial decision is made about the 'suitability' of a developing thymocyte to become a Treg cell. On the other hand, as Treg cells can recognize a wide range of foreign antigens, have a diverse TCR repertoire, and show some degree of overlap with Tconv cells, the signals through the TCR may be complementary to the TCR-independent process that generates precursors of Treg cells. In this review, we discuss how different features of the Treg repertoire influence our understanding of Treg specificities and the role of self reactivity in the generation of this population. PMID- 19128357 TI - Post-mortem analysis of islet pathology in type 1 diabetes illuminates the life and death of the beta cell. PMID- 19128358 TI - Clinical immunology review series: An approach to the management of pulmonary disease in primary antibody deficiency. AB - The sinopulmonary tract is the major site of infection in patients with primary antibody deficiency syndromes, and structural lung damage arising from repeated sepsis is a major determinant of morbidity and mortality. Patients with common variable immunodeficiency may, in addition, develop inflammatory lung disease, often associated with multi-system granulomatous disease. This review discusses the presentation and management of lung disease in patients with primary antibody deficiency. PMID- 19128359 TI - Analysis of islet inflammation in human type 1 diabetes. AB - The immunopathology of type 1 diabetes (T1D) has proved difficult to study in man because of the limited availability of appropriate samples, but we now report a detailed study charting the evolution of insulitis in human T1D. Pancreas samples removed post-mortem from 29 patients (mean age 11.7 years) with recent-onset T1D were analysed by immunohistochemistry. The cell types constituting the inflammatory infiltrate within islets (insulitis) were determined in parallel with islet insulin content. CD8(+) cytotoxic T cells were the most abundant population during insulitis. Macrophages (CD68(+)) were also present during both early and later insulitis, although in fewer numbers. CD20(+) cells were present in only small numbers in early insulitis but were recruited to islets as beta cell death progressed. CD138(+) plasma cells were infrequent at all stages of insulitis. CD4(+) cells were present in the islet infiltrate in all patients but were less abundant than CD8(+) or CD68(+) cells. Forkhead box protein P3(+) regulatory T cells were detected in the islets of only a single patient. Natural killer cells were detected rarely, even in heavily inflamed islets. The results suggest a defined sequence of immune cell recruitment in human T1D. They imply that both CD8(+) cytotoxic cells and macrophages may contribute to beta cell death during early insulitis. CD20(+) cells are recruited in greatest numbers during late insulitis, suggesting an increasing role for these cells as insulitis develops. Natural killer cells and forkhead box protein P3(+) T cells do not appear to be required for beta cell death. PMID- 19128361 TI - Neutrophil mobilization and clearance in the bone marrow. AB - The bone marrow is the site of neutrophil production, a process that is regulated by the cytokine granulocyte colony-stimulating factor (G-CSF). Mature neutrophils are continually released into the circulation, with an estimated 10(11) neutrophils exiting the bone marrow daily under basal conditions. These leucocytes have a short half-life in the blood of approximately 6.5 hr, and are subsequently destroyed in the spleen, liver and indeed the bone marrow itself. Additionally, mature neutrophils are retained in the bone marrow by the stromal cell-derived factor (SDF-1alpha)/chemokine (C-X-C motif) receptor 4 (CXCR4) chemokine axis and form the bone marrow reserve. Following infection or inflammatory insult, neutrophil release from the bone marrow reserve is substantially elevated and this process is mediated by the co-ordinated actions of cytokines and chemokines. In this review we discuss the factors and molecular mechanisms regulating the neutrophil mobilization and consider the mechanisms and functional significance of neutrophil clearance via the bone marrow. PMID- 19128364 TI - What are the keys to successful adrenal venous sampling (AVS) in patients with primary aldosteronism? AB - Adrenal venous sampling (AVS) is the criterion standard to distinguish between unilateral and bilateral adrenal disease in patients with primary aldosteronism. The keys to successful AVS include appropriate patient selection, careful patient preparation, focused technical expertise, defined protocol, and accurate data interpretation. The use of AVS should be based on patient preferences, patient age, clinical comorbidities, and the clinical probability of finding an aldosterone-producing adenoma. AVS is optimally performed in the fasting state in the morning. AVS is an intricate procedure because the right adrenal vein is small and may be difficult to locate - the success rate depends on the proficiency of the angiographer. The key factors that determine the successful catheterization of both adrenal veins are experience, dedication and repetition. With experience, and focusing the expertise to 1 or 2 radiologists at a referral centre, the AVS success rate can be as high as 96%. A centre-specific, written protocol is mandatory. The protocol should be developed by an interested group of endocrinologists, radiologists and laboratory personnel. Safeguards should be in place to prevent mislabelling of the blood tubes in the radiology suite and to prevent sample mix-up in the laboratory. PMID- 19128365 TI - Alterations in biomarkers of cardiovascular disease (CVD) in active acromegaly. AB - OBJECTIVES: In acromegalic patients, cardiovascular and metabolic comorbidities contribute to enhance mortality. Available data on the lipoprotein profile of these patients are controversial. Our aim was to characterize the lipoprotein profile and emergent biomarkers of cardiovascular disease in active acromegalic patients in comparison with sex- and age-matched healthy controls. PATIENTS: Eighteen patients with active acromegaly and 18 controls were studied. MEASUREMENTS: Glucose levels, hormonal status, lipoprotein profile and C reactive protein (CRP) were evaluated by standardized methods. Cholesteryl ester transfer protein (CETP) and lipoprotein-associated phospholipase A(2 )(Lp-PLA(2)) were measured by radiometric techniques, endothelin-1 and vascular cell adhesion molecule (VCAM)-1 by enzyme-linked immunosorbent assay, and leucocytes CD18, CD49d and CD54 by flow cytometry. RESULTS: After adjusting for body mass index (BMI), acromegalic patients presented a more atherogenic lipoprotein profile, consisting of higher levels of triglycerides and apolipoprotein B and alterations in the ratios which estimate insulin resistance and atherogenic risk. CETP activity was significantly increased in acromegalic patients as compared to controls (168 +/- 17 vs. 141 +/- 30% per ml h, respectively; P < 0.05). Endothelin-1 levels evidenced an increase in the patients' group (0.9 +/- 0.2 vs. 0.7 +/- 0.2 ng/l, respectively; P < 0.01) and showed positive and significant correlations with GH, IGF-1 and IGFBP-3 (r = 0.45, 0.42 and 0.44, respectively; P < 0.01 for all of them; with BMI as a fixed variable). Lymphocytes from acromegalic patients showed increased CD49d content (282 +/- 59 vs. 246 +/- 48 arbitrary units, respectively; P < 0.05). CONCLUSIONS: Taken together, the alterations described seem to contribute to constituting a state of higher propensity for the development of atherosclerotic cardiovascular disease, which adds to the presence of specific cardiomyopathy. PMID- 19128366 TI - Molecular analysis of novel PROP1 mutations associated with combined pituitary hormone deficiency (CPHD). AB - OBJECTIVE: Homozygous mutations in the gene encoding the pituitary transcription factor PROP1 are associated with combined pituitary hormone deficiency (CPHD) in both mice and humans with a highly variable phenotype with respect to the severity and time of initiation of pituitary hormone deficiency. We have ascertained three pedigrees with PROP1 mutations from a large cohort of patients with variable degrees of CPHD who were screened for mutations in PROP1. RESULTS: Affected individuals from all three pedigrees were found to harbour novel PROP1 mutations. We have identified two siblings in one family who were homozygous for an intronic mutation (c.343-11C > G) that disrupts correct splicing resulting in the loss of exon 3 from the PROP1 transcript. Two siblings from a second, unrelated family are compound heterozygotes for two point mutations in the coding region, a missense mutation (p.R125W) that leads to impaired transcriptional activation, and a deletion of a single nucleotide (c.310delC) resulting in a frameshift and nonfunctional mutant protein. Additionally, we identified a homozygous deletion of the PROP1 locus in two patients born to consanguineous parents. CONCLUSION: Mutations in PROP1 are a frequent cause of familial CPHD. We have described four novel mutations in PROP1 in 3 pedigrees, all resulting in PROP1 deficiency by different mechanisms. The phenotypic variation observed in association with PROP1 mutations both within and between families, together with the evolving nature of hormone deficiencies and sometimes changing pituitary morphology indicates a need for continual monitoring of these patients. PMID- 19128367 TI - Valvular heart disease and the use of cabergoline for the treatment of prolactinoma. AB - OBJECTIVE: The use of high doses of the ergot-derived dopamine agonist cabergoline (> 3 mg/day), especially with cumulative doses > 4000 mg, has been associated with an increase in cardiac valvular thickening and significant (moderate to severe) regurgitation. Whether lower doses commonly used in the treatment of prolactinomas (0.25-3 mg/week) are also associated with significant valvulopathy is controversial. The mitral valve tenting area, a subclinical index of leaflet stiffening, has also been correlated with the cumulative dose of cabergoline and severity of valvular regurgitation. DESIGN/PATIENTS/MEASUREMENTS: We performed transthoracic echocardiography (TTE) on 50 prolactinoma patients (48% macroprolactinomas, 52% microprolactinomas, 30 male, 20 female, age 51.2 +/- 2.2 years, mean +/- SEM) who had been taking cabergoline for 6.6 +/- 0.5 years (range 1-13 years) with cumulative doses of 443 +/- 53 mg, to determine the prevalence of significant valvular thickening (> 0.5 cm) and regurgitation, and measured the mitral valve tenting area and height. The results were compared to those from age- and sex-matched controls with normal left ventricular function. RESULTS: No significant valvular thickening or regurgitation of any valve was detected in the prolactinoma group and the prevalence of mild valvular regurgitation was not higher than in the case-control group. The mitral valve tenting area and height were not significantly greater than in the control group. There was no correlation between tenting area or height and cumulative cabergoline dose. CONCLUSIONS: We found no evidence of increased mitral valve tenting area/height, valvular thickening or significant regurgitation with the long-term administration of the commonly used doses of cabergoline to treat prolactinoma. PMID- 19128368 TI - Metformin maintains the weight loss and metabolic benefits following rimonabant treatment in obese women with polycystic ovary syndrome (PCOS). AB - OBJECTIVE: Rimonabant has been shown to reduce weight, free androgen index (FAI) and insulin resistance in obese patients with polycystic ovary syndrome (PCOS) compared to metformin. Studies have shown that significant weight regain occurs following the cessation of rimonabant therapy. This study was undertaken to determine if subsequent metformin treatment after rimonabant would maintain the improvement in weight, insulin resistance and hyperandrogenaemia in PCOS. DESIGN: An extension study for 3 months with the addition of metformin to the randomised open labelled parallel study of metformin and rimonabant in 20 patients with PCOS with a body mass index >or= 30 kg/m(2). Patients who were on 3 months of rimonabant were changed over to metformin for 3 months, whereas those on 3 months of metformin were continued on metformin for another 3 months. MEASUREMENTS: The primary end-point was a change in weight; secondary end-points were a change in FAI and insulin resistance. RESULTS: The mean weight loss of 6.2 kg associated with 3 months of rimonabant treatment was maintained by 3 months of metformin treatment (mean change +0.2 kg, P = 0.96). Therefore, the percentage reduction in weight remained significantly higher in the rimonabant/metformin group compared to metformin only subjects at 6 months compared to baseline (-6.0 +/- 0.1%vs. 2.8 +/- 0.1%, P = 0.04). The percentage change in testosterone and FAI from baseline to 6 months was also greater in the rimonabant/metformin group. [Testosterone (-45.0 +/- 5.0%vs. -16 +/- 2.0%, P = 0.02); FAI (-53.0 +/- 5.0%vs. 17.0 +/- 12.2%, P = 0.02)]. HOMA-IR continued to fall significantly in the rimonabant/metformin group between 0, 3 and 6 months (4.4 +/- 0.5 vs. 3.4 +/- 0.4 vs. 2.7 +/- 0.3, respectively, P < 0.01) but not at all in the metformin only group (3.4 +/- 0.7 vs. 3.4 +/- 0.8 vs. 3.7 +/- 0.8, respectively, P = 0.80). Total cholesterol and LDL reduced significantly in both groups, but improvements in triglycerides and HDL were limited to the rimonabant/metformin group. CONCLUSIONS: In these obese patients with PCOS, metformin maintained the weight loss and enhanced the metabolic and biochemical parameters achieved by treatment with rimonabant, compared to 6 months of metformin treatment alone. PMID- 19128369 TI - Abstracts of the ISPOR (International Society for Pharmacoeconomics and Outcomes Research) Eleventh Annual European Congress. November 8-11 2008, Athens, Greece. PMID- 19128370 TI - Long-term outcomes of trabeculectomy. PMID- 19128371 TI - Endothelial keratoplasty: where are we? PMID- 19128372 TI - Horizontal skin wound closure in full thickness eyelid excision. AB - The direct closure of full thickness lid margin defects is well described. The most popular method of closing this defect is to convert it to a pentagon by excising a triangle of skin and muscle below. The resultant scar is perpendicular to relaxed skin tension lines and may be cosmetically obvious. An alternative technique is presented whereby a horizontal subciliary incision is extended either side of the defect. After undermining and minimal excision the skin is closed leaving a horizontal linear subciliary scar, parallel to relaxed skin tension lines. PMID- 19128373 TI - Trans-vitreal endoresection for vasoproliferative retinal tumours. AB - We present an interventional consecutive case series to describe our experience of transvitreal endoresection (TVE) for vasoproliferative retinal tumours (VPRTs). Three patients with VPRTs refractory to conventional treatment modalities of cryotherapy, plaque radiotherapy and anti-VEGF presented with macular exudative changes and were offered TVE. Complete ophthalmic examination with colour fundal photographs was performed before and during the follow-up period. All patients were followed up for 6 months following silicone oil removal. At the last follow up, resolution of macular and retinal exudative changes with parallel improvement in vision was observed. No recurrences were detected during the follow-up period. Two of three patients had simultaneous cataract surgery and developed fibrinous uveitis, requiring management with intensive topical steroids. After 3 months, these patients had a fibrotic pupillary membrane enveloping the intraocular lens (IOL) with posterior synechiae. These two patients underwent SO removal, IOL explantation and artisan IOL (iris clipped) insertion, resulting in visual improvement. We did not observe any PVR complications in our cases. TVE represents an effective and safe option for the treatment of VPRTs refractory to other treatment modalities. Further studies with a larger sample size and long-term follow up are indicated to evaluate the role of TVE in the management of VPRTs. PMID- 19128374 TI - Serum growth factor analysis in dry eye syndrome. AB - BACKGROUND: To perform a comprehensive serum growth factor analysis in dry eye syndrome patients and to compare this with matched controls. METHODS: Six female dry eye syndrome patients and six age- and gender-matched controls were recruited. Whole blood was collected, allowed to clot and then centrifuged. Serum was extracted by using sterile technique. Enzyme-linked immunosorbent assays were performed to quantify serum growth factor levels. RESULTS: Levels of transforming growth factor-beta 1 and 2 (TGF-beta1 and beta2), nerve growth factor (NGF), insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), acidic and basic fibroblast growth factor (FGF), keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor-AA, AB and BB (PDGF-AA, AB and BB), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and glial cell line-derived neurotrophic factor (GDNF) were quantified, and statistical analysis was performed by using the Mann-Whitney U-test with the Bonferroni correction. CONCLUSIONS: No significant difference was found between serum growth factor levels in dry eye syndrome patients versus controls. Our study provides comprehensive analysis of serum growth factor levels in autologous serum eye drops produced from ocular surface disease patients. A knowledge of growth factor levels in serum may be important because of the increasing use of autologous serum eye drops in refractory ocular surface diseases and for an understanding of how topical serum may provide benefit. PMID- 19128375 TI - Fungal keratitis in far north Queensland, Australia. AB - BACKGROUND: Aboriginals and Torres Strait Islanders form a significant proportion of the population in far north Queensland and are at increased risk for many eye conditions. This study describes the pattern of fungal keratitis seen at Cairns Base Hospital, far north Queensland, Australia. METHODS: A retrospective review of all cases of culture positive fungal keratitis presenting between 1998 and 2008. The records of 17 patients were reviewed for epidemiological, risk factor, microbiological, treatment and outcome data. RESULTS: The study included 16 eyes from 16 patients. Five (31.25%) patients were of Aboriginal and Torres Strait Islander descent. Pre-disposing factors were ocular trauma (n = 7) and contact lens wear (n = 3). No patients were on topical steroids at presentation. All patients grew filamentous fungi with Fusarium the most common isolate (50%). Aspergillus, Curvularia and Lasiodiplodia theobromae were the next most common accounting for two cases each. Two patients developed corneal perforations, whereas two required penetrating keratoplasty and one required evisceration. Eight patients had a visual acuity of 6/18 or better at presentation and this increased to 13 patients at final follow up. CONCLUSIONS: A significant proportion of the patients presenting to Cairns Base Hospital with fungal keratitis are Indigenous. The very high percentage of cases due to filamentous fungi is similar to other tropical regions of the world. The very low rates of Candida infection and steroid use prior to presentation are in contrast to studies from temperate areas such as Melbourne and Philadelphia. PMID- 19128376 TI - Rapid visual recovery after penetrating keratoplasty for keratoconus. AB - PURPOSE: To ascertain the level and speed of visual recovery after penetrating keratoplasty for keratoconus. METHOD: A retrospective review was performed of 100 consecutive cases of penetrating keratoplasty for keratoconus, performed between 1999 and 2005. Review assessed visual function and the speed at which patients achieved a functional best corrected visual acuity (BCVA) of > or = 6/12 either with glasses or phoropter. Analysis of visual, refractive and keratometric results were made on 76 eyes that had reached 6 months after suture removal. Intraoperative and postoperative complications including graft rejections were recorded. RESULTS: Post keratoplasty, 43.4%, 78.9% and 96.1% of patients achieved a BCVA of 6/12 or better by 1, 3 and 6 months, respectively. The mean time to achieve a BCVA of 6/12 was 9.6 weeks. Only 5.3% of patients required a rigid gas permeable contact lens. Overall 42.4% of patients had unaided visual acuity of 6/12 or better at 12 months. The mean refractive cylinder and standard deviation was 2.78 +/- 1.6 D, and the mean spherical equivalent was -1.12 +/- 2.9 D. There were no significant intraoperative complications, and although 3.9% of eyes had at least one graft rejection episode there were no graft failures. CONCLUSION: Penetrating keratoplasty is an effective method for treating advanced keratoconus. By 3 months almost all patients are able to achieve a BCVA of 6/12 or better with progressive improvement over time. This technique allows almost half of patients to achieve an unaided vision of 6/12 or better with sutures in situ. PMID- 19128377 TI - Otago Glaucoma Surgery Outcome Study: long-term results of 841 trabeculectomies. AB - BACKGROUND: To describe the long-term outcomes of trabeculectomies performed at Dunedin Hospital and followed in the Otago Glaucoma Surgery Outcome Study. METHODS: Prospective non-comparative case series of 841 eyes of 607 patients who had first trabeculectomies for primary open- or closed-angle glaucoma at Dunedin Hospital between 1976 and 2005 and followed for a mean of 7.5 years (standard deviation 6.0). RESULTS: The probability of a trabeculectomy controlling the intraocular pressure at 21 mmHg or less at 1, 10 and 20 years was 0.96 (95% confidence interval [CI] 0.95, 0.97), 0.86 (95% CI 0.83, 0.89) and 0.79 (95% CI 0.74, 0.83), respectively. Visual acuity was maintained or improved between preoperative assessment and final follow up in 68% of cases. The probability of not being blind following trabeculectomy at 1, 10 and 20 years was 0.98 (95% CI 0.96, 0.98), 0.83 (95% CI 0.80, 0.87) and 0.70 (95% CI 0.64, 0.76), respectively. The proportion of those with glaucomatous field loss increased during follow up from 16% (44/283) at 0-5 years to 50% (10/20) for those with 21 or more years of follow up. A repeat drainage procedure was required in 65 eyes (8%) (56 Molteno implant insertions and 9 repeat trabeculectomies). CONCLUSIONS: Intraocular pressure was well controlled by trabeculectomy; however, a steady decline in intraocular pressure control, visual acuity and visual field occurred during follow up. PMID- 19128378 TI - Blood flow in the peripapillary retina in exfoliation glaucoma. AB - PURPOSE: The aim of this article is to study peripapillary retinal blood flow in patients with progressive and stable exfoliation glaucoma (ExG). METHODS: Fifty eight eyes with ExG were included; 25 of them had progressive and 33 stable glaucoma. Retinal blood flow in the peripapillary retina was measured with scanning laser Doppler flowmetry. Acquired flow maps were analysed with the automatic full-field perfusion image analyser. Multiple logistic regression was used to model progression of glaucoma. RESULTS: Mean retinal flow (MF; correlation coefficient, P-value; R = 0.36, P = 0.006) and retinal minimum diastolic flow (R = 0.33, P = 0.011) were positively correlated with visual field mean defect (MD). Factors associated with progressive glaucoma were mean intraocular pressure (OR = 1.198 for each mmHg; P = 0.050) and visual field MD (OR = 1.134 for each dB; P = 0.013). Age (P = 0.35), MF (P = 0.58), or presence of cardiovascular disease (P = 0.17) were not associated with glaucoma progression. CONCLUSION: No difference in peripapillary retinal blood flow between progressive and stable ExG could be found. PMID- 19128379 TI - Aggregations of retinal pigment epithelial cells on inferior retinal blood vessels, a clinical sign of early proliferative vitreoretinopathy. AB - PURPOSE: To determine the relationship between 'pigment clusters' on blood vessels (BvRPE) on the inferior retina in patients with rhegmatogenous retinal detachment, and existing proliferative vitreoretinopathy (PVR). METHODS: A prospective observational non-comparative study of consecutive patients presenting with rhegmatogenous retinal detachment to a tertiary referral university department was performed. The retina was inspected preoperatively for the detection of BvRPE. A relationship with existing PVR was determined. RESULTS: A total of 103 patients were recruited, 70 men and 32 women, mean age 56.7 years, with 57 left eyes. Mean duration of follow up was 9.2 months. Nineteen patients (18.4%) had BvRPE, and 20 had RPE clusters off blood vessels (nine had both). In those with the sign of BvRPE the mean number of RPE clusters on blood vessels was 5.7 (SD 5.8). The mean number of clock hours affected was 2.32 (SD = 1.58). Eight had B grade PVR and three C grade PVR. Patients with BvRPE were more likely to have PVR grades B or C than those without the sign (P = 0.002). The mean number of operations in those with the sign was 1.32 (SD = 0.58) and was statistically similar to those without BvRPE. Visual outcome was statistically similar in those with and without the sign. CONCLUSION: Patients with rhegmatogenous retinal detachment may demonstrate 'pigment clusters' on blood vessels in the inferior retina (BvRPE). This is likely to be a specific feature found in PVR and an early sign of activation of retinal pigment epithelial cells. PMID- 19128380 TI - Intravitreal bevacizumab (Avastin) for neovascular age-related macular degeneration using a variable frequency regimen in eyes with no previous treatment. AB - PURPOSE: To evaluate a variable frequency regimen with intravitreal bevacizumab for treatment of neovascular age-related macular degeneration (AMD) in eyes that have not received any previous treatment. METHODS: Retrospective review of patients with neovascular AMD who were treated with three consecutive monthly intravitreal injections of bevacizumab (1.25 mg) and retreated based on the PrONTO study criteria. Outcome measures included visual acuity (VA) and central retinal thickness. Subgroup analysis was conducted to identify pretreatment characteristics that could determine visual outcome with treatment. RESULTS: A total of 109 eyes of 109 patients were treated. The mean age was 82 years, and the mean follow-up period was 9.4 months (range 6-12 months). At baseline, the mean VA was 45.6 letters (6/37.5) and mean central retinal thickness 343 microm. This improved to 51 letters (6/30) (P < 0.001)) and 231 microm (P < 0.001) at 6 months. At 6 months, VA was improved by at least five letters in 50%, remained stable in 30% and worsened by at least five letters in 20% of patients. Patients with large intraretinal cysts on optical coherence tomography before treatment had an increased risk of worse vision (odds ratio 10.5, 95% confidence interval 1.69-64.99; P = 0.018). CONCLUSIONS: The majority of patients had improvement or stability of VA regardless of the angiographic type of choroidal neovascularization. Intravitreal bevacizumab with this tailored regimen is beneficial in the treatment of neovascular AMD in the short term. The presence of large intraretinal cysts on optical coherence tomography is a poor prognostic factor for visual improvement with this treatment. PMID- 19128381 TI - Scleral wrap increases the long-term complication risk of bone-derived hydroxyapatite orbital implants. AB - BACKGROUND: This study was prompted by previous findings that suggested that scleral wrapping increased the rate of complications following insertion of bone derived hydroxyapatite orbital implants and compared the long-term results of implants inserted with and without scleral wraps. METHODS: This retrospective case series reviewed the long-term outcomes of 159 patients who had undergone enucleation with insertion of a bone-derived hydroxyapatite orbital implant at Dunedin Hospital between 1977 and 2006. Implants were inserted with and without scleral wraps in 85 and 74 cases, respectively. Follow up was 0.5-27.5 years (mean 8.2 years) for the whole series and 9.7 years for the sclera group and 6.7 years for the group without sclera. Patient details were obtained from theatre records, case note review, patient interview and examination, interview of patient relatives and family general practitioner records. The main outcome measures were the rates of minor or major complications and their treatments and outcomes. RESULTS: Twenty-seven cases (17%) suffered minor complications of limited implant exposure that either healed spontaneously, with implant drilling or wound resuturing and 11 cases (7%) suffered major complications requiring explantation. Of 38 patients with postoperative complications, 31 (82%) had scleral wraps compared with 7 (18%) without sclera (P < 0.001; OR 5.14, 95% CI 2.00-14.78). CONCLUSION: Bone-derived hydroxyapatite orbital implants inserted without scleral wrap were associated with better clinical outcomes and a lower rate of long-term complications. It is therefore recommended that these implants be inserted without a scleral wrap. PMID- 19128382 TI - The New Zealand ophthalmology workforce 2008. AB - BACKGROUND: The ongoing supply of quality ophthalmic care to the New Zealand (NZ) population depends on a sustainable ophthalmology workforce. The NZ population is growing and aging rapidly, placing increasing demands on ophthalmic services. Predicting future supply of NZ ophthalmologists requires an accurate description of the current workforce. METHODS: A confidential postal workforce survey, the fourth since 1997, was sent to all NZ consultant ophthalmologists in October 2007. A second survey conducted in January 2008 by email and direct telephone contact gained a complete dataset of NZ ophthalmologists. Results were compared with NZ population data, and the Australian Ophthalmology Workforce Model 2006 was used to estimate future demands on NZ ophthalmic services. RESULTS: There are currently 16-24 vacancies for consultant ophthalmologists in NZ, with another eight positions planned to be created by 2010, bringing the shortfall to 24-32. Retirement rates average to three per year, although 28 consultants are planning to retire by 2013. Retention of new NZ Fellows stands at around 71%. Increasing demands of an aging population are likely to increase the shortfall of ophthalmologists to 40-50 by 2015, and 46-69 by 2025. CONCLUSIONS: This report identifies a growing gap between supply of NZ ophthalmologists and demand for their services. We highlight the need for a comprehensive model with a detailed analysis of the many factors influencing supply and demand, enabling planning for an appropriate supply of ophthalmologists meeting the needs of the future NZ population. PMID- 19128383 TI - Local effect of topical FP-receptor agonists on retinal vessels of the ipsilateral posterior retina in normal rabbit eyes. AB - PURPOSE: To determine whether a topically instilled prostaglandin analogue inhibits endothelin-1 (ET-1)-induced vasoconstrictive effects in the posterior retina by its local effects, and the duration of the effect in normal rabbit eyes. METHODS: Travoprost, a potent selective FP-agonist, or unoprostone, a prostone that also has a weak non-selective FP-agonistic activity, solution was instilled once, or once daily (travoprost) or twice daily (unoprostone) for 7 days in one randomly chosen Dutch rabbit eye, and vehicle in the contralateral eye. ET-1 was intravitreously injected in both eyes 30 min after a single instillation of a test drug or its vehicle, or just after, 30, 60, 90 or 180 min after the final instillation of a 7-day instillation, and fundus photographed before, 30 and 60 min after the injection to study whether difference was seen in the ET-1-induced constriction of retinal vessels between the drug- and vehicle instilled eyes. The same experimental procedures were conducted with indomethacin pretreatment. RESULTS: In the rabbit eyes where travoprost was instilled for 7 days, the ET-1-induced constriction of retinal vessels was significantly inhibited only on the drug-treated side, when ET-1 was injected 30 or 60 min after the final instillation (P = 0.026-0.005), which was abolished by indomethacin pretreatment. A single instillation of travoprost or unoprostone and 7-day instillation of unoprostone showed no effect. CONCLUSIONS: After a 7-day instillation in normal rabbit eyes, topical travoprost suppressed ET-1-induced vasoconstrictive effects only in the ipsilateral posterior retina by its local effect; this effect was maintained at least for 30 min and mediated by endogenous prostaglandins. PMID- 19128384 TI - Life-threatening conjunctival presentation of myeloid sarcoma. AB - Myeloid sarcoma is a life-threatening condition that can present rarely as a conjunctival lesion. This case report describes a 48-year-old woman who was diagnosed after conjunctival biopsy, subsequently systemically investigated and then treated with chemotherapy. Collaboration of the ophthalmologist, pathologist and oncologist was critical in the successful management of this patient. PMID- 19128385 TI - Clinicopathological features of ocular cystinosis. AB - A 31-year-old woman who presented with photophobia was found to have bilateral corneal and conjunctival crystal deposition. Ocular cystinosis was diagnosed upon observation of typical crystals and lack of systemic involvement. In vivo confocal microscopy confirmed crystal deposition of the corneas and conjunctivae bilaterally. Optical coherence tomography showed stromal hyper-reflectivity due to crystals within the corneal stroma. Transmission electron microscopy of the conjunctiva demonstrated pathognomonic intralysosomal cystine crystals inside fibroblasts and macrophages. Clinicopathological characteristics of ocular cystinosis are well described by this exceptional case. PMID- 19128386 TI - Removal of internal limiting membrane for recurrent myopic traction maculopathy. AB - A 63-year-old man presented with a foveal detachment and retinoschisis associated with myopic traction maculopathy of his left eye with a refractive error of -12.0 dioptres. Both the retinoschisis and foveal detachment were initially successfully treated with vitreous surgery that was limited to the induction of a posterior vitreous detachment beyond the areas of retinoschisis over the posterior staphyloma. Three years later, the macula re-detached and further surgery was performed, which involved peeling the internal limiting membrane (ILM). Following the second vitreous surgery the macula reattached within one month of the surgery and this success has been sustained for 2 years, suggesting that peeling the ILM may be an effective treatment for recurrent myopic traction maculopathy. PMID- 19128387 TI - Conjunctival melanoma and melanosis: a reappraisal of terminology, classification and staging. AB - This paper aims to stimulate debate on the terminology, classification, grading and staging of conjunctival melanosis and melanoma. We audited our results with 76 invasive conjunctival melanomas. Staging according to the sixth edition of the Tumour Node Metastasis (TNM) system did not correlate well with tumour extent and outcome. Approximately 50% of invasive melanomas were associated with 'primary acquired melanosis with atypia', a term which in our opinion underestimates the gravity of this disease. We also found deficiencies in the grading, terminology and classification of conjunctival melanocytic abnormalities. In summary, we suggest that the term 'primary acquired melanosis' be reserved for clinical diagnosis. Histologically, this abnormality can be categorized more precisely as either 'hypermelanosis' or 'conjunctival melanocytic intraepithelial neoplasia (C MIN)'. 'Primary acquired melanosis without atypia' can be termed more accurately as 'C-MIN without atypia'. In view of the high risk of invasive melanoma, we suggest that 'primary acquired melanosis with atypia' be termed 'C-MIN' with atypia, with the more severe changes regarded as melanoma in situ. To improve objectivity in the reporting of C-MIN, we propose a scoring system based on horizontal and vertical spread and degree of severity of melanocytic atypia. We suggest that the TNM staging system for conjunctival melanoma be revised to: (i) include a Tis stage; (ii) take account of tumour size, quadrant and caruncular involvement; and (iii) improve staging of any local invasion beyond conjunctiva. PMID- 19128388 TI - Anterior segment optical coherence tomography in the investigation of an unusual case of entoptic phenomenon. PMID- 19128389 TI - Q fever-associated HLAB27 anterior uveitis. PMID- 19128390 TI - Bilateral spontaneous rupture of posterior capsule in posterior polar cataract. PMID- 19128391 TI - Localized bullous keratopathy secondary to posterior polymorphous dystrophy. PMID- 19128392 TI - Chronic postoperative endophthalmitis with isolation of Leuconostoc and Olsenella uli in an Indigenous man. PMID- 19128393 TI - Impaired cerebrospinal fluid circulation and its relationship to glaucoma. PMID- 19128396 TI - A diagnostic tattoo. AB - The hallmarks of the myoclonic epilepsy with ragged red fibers (MERRF) syndrome are myoclonic epilepsy, ataxia and ragged red fibres detected on muscle biopsy. We present a case of a 25-year-old male who first presented to his general practitioner at the age of 22 years with myoclonic jerks affecting the arms and legs, fatigue and mild ataxia. He was found to carry an A>G transition at nucleotide 8344 in mitochondrial DNA. This mutation is the most common cause of the MERRF syndrome, found in more than 80% of affected patients. Our patient had the diagnosis tattooed on his arm, both out of frustration at how few people had heard of it, and as a way of accepting that his condition was a part of who he was. Although the MERRF syndrome is one of the more common forms of mitochondrial encephalomyopathy, with a prevalence estimated at between 0.25 and 0.39 per 100,000, it is still a rare disorder. We are always striving to increase the public's understanding of these important conditions. Our patient has perhaps helped more than most towards this aspiration. PMID- 19128400 TI - Takahashi's response to Ravn's correspondence. PMID- 19128401 TI - Expression of cardiac myosin light chain 2 during embryonic heart development in medaka fish, Oryzias latipes, and phylogenetic relationship with other myosin light chains. AB - Cardiac myosin light chain 2 (MLC-2) plays a key role in heart development, contraction, and embryo and adult heart maintenance. In some animals, defects in the function of cardiac MLC-2 cause hypertrophic cardiomyopathy. To illuminate the functions of cardiac MLC-2 in embryonic heart formation and contraction, and into the evolution of MLC-2, we characterized the expression and requirement for medaka cardiac MLC-2 gene in the developing heart. Medaka cardiac MLC-2 cDNA (mcmlc2) was isolated and its gene expression pattern was determined. The mcmlc2 was found to be expressed in the bilateral cardiac mesoderm, the formed heart tube, and in both the differentiated ventricle and atrium. Knockdown of mcmlc2 function caused severe cardiac disorders, including edema in the atrium and sinus venosus. Using phylogenetic analysis, we found that physiological variations in the MLC-2 molecules evolved due to amino acid changes in the Ca(2+) binding domain during molecular evolution. Our findings concerning the function and expression of mcmlc2 are nearly identical with those of other MLC-2 genes, and our phylogenetic analysis suggests that during evolution, the variations in physiological function within the MLC-2 gene family have arisen from a change in the amino acids in the Ca(2+) binding domain in the MLC-2 molecule. PMID- 19128402 TI - Absence of tissue inhibitor of metalloproteinases 3 disrupts alveologenesis in the mouse. AB - Tissue inhibitors of metalloproteinases (TIMPs) regulate extracellular matrix (ECM) degradation by matrix metalloproteinases (MMPs) throughout lung development. We examined lungs from TIMP3 null mice and found significant air space enlargement compared with wild type (WT) animals during a time course spanning early alveologenesis (post-partum days 1, 5, 9 and 14). Trichrome staining revealed a similar pattern of collagen distribution in the walls of nascent alveoli; however, the alveolar walls of TIMP3 mutant mice appeared to be thinner than controls. Assessment of MMP2 and MMP9 activities by gelatin zymography demonstrated a significant elevation in the active form of MMP2 at post-partum days 1 and 5. Treatment of null pregnant dams with a broad spectrum synthetic metalloproteinase inhibitor, GM6001, on embryonic day 16.5 enhanced the formation of primitive alveoli during the saccular stage of lung development as evidenced by a partial, but significant, rescue of alveolar size in post-partum day 1 animals. We propose that increased MMP activity in the absence of TIMP3 enhances ECM proteolysis, upsetting proper formation of primitive alveolar septa during the saccular stage of alveologenesis. Therefore, TIMP3 indirectly regulates alveolar formation in the mouse. To our knowledge, ours is the first study to demonstrate that in utero manipulation of the TIMP/MMP proteolytic axis, to specifically inhibit proteolysis, significantly affects lung development. PMID- 19128403 TI - Spatio-temporal control of neural epithelial cell migration and epithelium-to mesenchyme transition during avian neural tube development. AB - As opposed to the neural crest, the neural epithelium is generally viewed as a static and cohesive structure. Here, using an ex vivo system free of the environmental influences and physical constraints encountered in the embryo, we show that neural epithelial cells are on the contrary intrinsically motile, although they do not undergo spontaneous epithelium-to-mesenchyme transition and display molecular and cellular characteristics distinct from those of neural crest cells. However, they can be instructed to undergo epithelium-to-mesenchyme conversion independently of the acquisition of neural crest traits. Migration potentialities of neural epithelial cells are transient and are progressively restricted during neural tube development. Restriction of cell migration is irreversible and can be in part accounted for by increase in N-cadherin in cellular junctions and in cell polarity. In conclusion, our study reveals that the neural epithelium is a highly flexible tissue in which cells are maintained cohesive under the control of a combination of extrinsic factors and physical constraints. PMID- 19128404 TI - Vocal control area-related expression of neuropilin-1, plexin-A4, and the ligand semaphorin-3A has implications for the evolution of the avian vocal system. AB - The avian vocal system is a good model for exploring the molecular basis of neural circuit evolution related to behavioral diversity. Previously, we conducted a comparative gene expression analysis among two different families of vocal learner, the Bengalese finch (Lonchura striata var. domestica), a songbird, and the budgerigar (Melopsittacus undulatus), a parrot; and a non-learner, the quail (Coturnix coturnix), to identify various axon guidance molecules such as cadherin and neuropilin-1 as vocal control area-related genes. Here, we continue with this study and examine the expression of neuropilin and related genes in these species in more detail. We found that neuropilin-1 and its coreceptor, plexin-A4, were expressed in several vocal control areas in both Bengalese finch and budgerigar brains. In addition, semaphorin-3A, the ligand of neuropilin-1, expression was not detected in vocal control areas in both species. Furthermore, there was some similar gene expression in the quail brain. These results suggest the possibility that a change in the expression of a combination of semaphorin/neuropilin/plexin was involved in the acquisition of vocal learning ability during evolution. PMID- 19128405 TI - Role of CCN, a vertebrate specific gene family, in development. AB - The CCN family of genes constitutes six members of small secreted cysteine rich proteins, which exists only in vertebrates. The major members of CCN are CCN1 (Cyr61), CCN2 (CTGF), and CCN3 (Nov). CCN4, CCN5, and CCN6 were formerly reported to be in the Wisp family, but they are now integrated into CCN due to the resemblance of their four principal modules: insulin like growth factor binding protein, von Willebrand factor type C, thrombospondin type 1, and carboxy terminal domain. CCNs show a wide and highly variable expression pattern in adult and in embryonic tissues, but most studies have focused on their principal role in osteo/chondrogenesis and vasculo/angiogenesis from the aspect of migration, growth, and differentiation of mesenchymal cells. CCN proteins simultaneously integrate and modulate the signals of integrins, bone morphogenetic protein, vascular endothelial growth factor, Wnt, and Notch by direct binding. However, the priority in the use of the signals is different depending on the cell status. Even the equivalent counterparts show a difference in signal usage among species. It may be that the evolution of the CCN family continues to keep pace with vertebrate evolution itself. PMID- 19128417 TI - Prevalence of ALDH7A1 mutations in 18 North American pyridoxine-dependent seizure (PDS) patients. AB - PURPOSE: Pyridoxine-dependent seizure (PDS) is a rare disorder characterized by seizures that are resistant to common anticonvulsants, and that are ultimately controlled by daily pharmacologic doses of pyridoxine (vitamin B6). Mutations of the antiquitin gene (ALDH7A1) are now recognized as the molecular basis of cases of neonatal-onset PDS. METHODS: Bidirectional DNA sequence analysis of ALDH7A1 was undertaken along with plasma pipecolic acid (PA) measurements to determine the prevalence of ALDH7A1 mutations in a cohort of 18 North American patients with PDS. RESULTS: In patients with neonatal-onset PDS, compound heterozygous or homozygous ALDH7A1 mutations were detected in 10 of 12 cases, and a single mutation was found in the remaining 2. In later-onset cases, mutations in ALDH7A1 were detected in three of six cases. In two patients with infantile spasms responsive to pyridoxine treatment and with good clinical outcomes, no mutations were found and PA levels were normal. In total, 13 novel mutations were identified. DISCUSSION: Our study advances previous findings that defects of ALDH7A1 are almost always the cause of neonatal-onset PDS and that defects in this gene are also responsible for some but not all later-onset cases. Later onset cases of infantile spasms with good outcomes lacked evidence for antiquitin dysfunction, suggesting that this phenotype is less compelling for PDS. PMID- 19128418 TI - Abstract of the Seventeenth Annual Computational Neuroscience Meeting: CNS*2008. Portland, Oregon, USA. July 19-20, 2008. PMID- 19128419 TI - Abstracts of the 14th Scientific Symposium of the Austrian Pharmacological Society (APHAR). Innsbruck, Austria. 21-22 November 21-22, 2008. PMID- 19128420 TI - Abstracts of the 11th Annual Society for Cardiovascular Magnetic Resonance (SCMR) Scientific Sessions. Los Angeles, California, USA. February 1-3, 2008. PMID- 19128421 TI - Abstracts of Symposium Mammographicum 2008. July 6-8, 2008. Lille, France. PMID- 19128422 TI - Controversies in Breast Cancer 2008. PMID- 19128423 TI - Assessing individual risk for breast cancer: role of oestrogens and androgens. PMID- 19128424 TI - Should lifestyle modifications be promoted to prevent breast cancer? PMID- 19128425 TI - Identification of personal risk of breast cancer: genetics. PMID- 19128426 TI - Assessing risk for breast cancer. PMID- 19128427 TI - Psychosocial aspects of risk appraisal. PMID- 19128428 TI - Introduction to sessions on 'Endocrine therapy: where have we come from, where are we at and where are we going to'? PMID- 19128429 TI - By looking back we can see the way forward: enhancing the gains achieved with antihormone therapy. PMID- 19128430 TI - Pharmacogenetics and pharmacogenomics of endocrine agents for breast cancer. PMID- 19128431 TI - Markers of endocrine sensitivity. PMID- 19128432 TI - Identification and mechanisms of endocrine resistance. PMID- 19128433 TI - Introduction to sessions on 'What can surgeons do for us (and we for them): radiology, pathology, molecular biology and local therapy'. PMID- 19128434 TI - Integration of endocrine therapy with targeted agents. PMID- 19128435 TI - Introduction to sessions on 'Optimizing the implementation of future treatments- time to abandon traditional methods'. PMID- 19128436 TI - Strength and weakness of phase I to IV trials, with an emphasis on translational aspects. AB - Although phase I to III trials represent the standard for introducing new drugs to clinical therapy, there has been increasing demand for translational research in oncology over the past decade. Thus, for most novel therapies such as 'targeted agents', a critical aspect for drug development in oncology has been to select the right patients for therapy. Translational research plays a pivotal role, not only in phase II trials but also in phase I and III and even in phase IV trials. The importance of distinguishing between our translational 'aims' in phase II and phase III trials is emphasized. Although translational research in phase III trials aims to identify optimal markers for clinical use, phase II studies may represent an optimal setting to explore tumour biology and the mechanisms of drug resistance in depth. PMID- 19128437 TI - Who should participate in clinical trials and who not? Can clinical trials be made more efficient and effective? PMID- 19128438 TI - Neoadjuvant/presurgical treatments. PMID- 19128439 TI - Evaluation of biological agents targeted at early-stage disease. PMID- 19128440 TI - Optimizing the implementation of future treatment using surrogate end-points. PMID- 19128441 TI - Thirty years of experience with mammography screening: a new approach to the diagnosis and treatment of breast cancer. PMID- 19128442 TI - The utility of conventional and molecular pathology in managing breast cancer. PMID- 19128443 TI - How much can improved molecular and pathologic discriminants change local therapy? PMID- 19128444 TI - Introduction to the sessions on 'Debate: extended follow-up of breast cancer patients in clinic wastes time for both patients and doctors'. PMID- 19128445 TI - Extended follow-up of breast cancer patients in clinic wastes time for both patients and doctors: the case for. PMID- 19128446 TI - Extended follow-up of breast cancer patients in clinic wastes time for both patients and doctors: the case against. PMID- 19128447 TI - Introduction to sessions on 'Predicting personal risk for breast cancer'. PMID- 19128448 TI - Pathways into homelessness: recently homeless adults problems and service use before and after becoming homeless in Amsterdam. AB - BACKGROUND: To improve homelessness prevention practice, we met with recently homeless adults, to explore their pathways into homelessness, problems and service use, before and after becoming homeless. METHODS: Recently homeless adults (last housing lost up to two years ago and legally staying in the Netherlands) were sampled in the streets, day centres and overnight shelters in Amsterdam. In April and May 2004, students conducted interviews and collected data on demographics, self reported pathways into homelessness, social and medical problems, and service use, before and after becoming homeless. RESULTS: among 120 recently homeless adults, (male 88%, Dutch 50%, average age 38 years, mean duration of homelessness 23 weeks), the main reported pathways into homelessness were evictions 38%, relationship problems 35%, prison 6% and other reasons 22%. Compared to the relationship group, the eviction group was slightly older (average age 39.6 versus 35.5 years; p = 0.08), belonged more often to a migrant group (p = 0.025), and reported more living single (p < 0,001), more financial debts (p = 0.009), more alcohol problems (p = 0.048) and more contacts with debt control services (p = 0.009). The relationship group reported more domestic conflicts (p < 0.001) and tended to report more drug (cocaine) problems. Before homelessness, in the total group, contacts with any social service were 38% and with any medical service 27%. Despite these contacts they did not keep their house. During homelessness only contacts with social work and benefit agencies increased, contacts with medical services remained low. CONCLUSION: the recently homeless fit the overall profile of the homeless population in Amsterdam: single (Dutch) men, around 40 years, with a mix of financial debts, addiction, mental and/or physical health problems. Contacts with services were fragmented and did not prevent homelessness. For homelessness prevention, systematic and outreach social medical care before and during homelessness should be provided. PMID- 19128449 TI - Efficiency of complex production in changing environment. AB - BACKGROUND: Cell function necessitates the assemblage of proteins into complexes, a process which requires further regulation on top of the fairly understood mechanisms used to control the transcription and translation of a single protein. However, not much is known about how protein levels are controlled to realize that regulation. RESULTS: We integrated data on the composition of yeast protein complexes and the dynamics of their protein building-blocks concentrations to show how the cell regulates protein levels to optimize complex formation. We find that proteins which are subunits of the same complex tend to have similar levels which change similarly following a change in growth conditions, and that abundant proteins undergo larger decrease in their copy number when grown in minimal media. We also study the fluctuations in protein levels and find them to be significantly smaller in large complexes, and in the least abundant subunit of each complex. We use a mathematical model of complex synthesis to explain how all these observations increase the efficiency of complex synthesis, in terms of better utilization of the available molecules and better resilience to stochastic variations. CONCLUSION: In conclusion, these results indicate an intricate regulation at all levels of protein production for the purpose of optimizing complex formation. PMID- 19128450 TI - Characterization of a prenatally assessed de novo supernumerary minute ring chromosome 20 in a phenotypically normal male. AB - BACKGROUND: The heterogeneous group of small supernumerary marker chromosomes (sSMCs) presents serious counseling problems, especially if they are present de novo and diagnosed prenatally. The incidence has been estimated at 1 in 1000 prenatal samples. We present a case of mosaic sSMC diagnosed prenatally after amniocentesis. The sSMC was characterized by various molecular cytogenetic techniques and determined to be a r(20) chromosome. After genetic counseling, the parents decided to continue the pregnancy, and a boy with minor phenotypic variants was born after 39 weeks of pregnancy. The case is compared with four other cases of prenatally detected r(20) mosaicism. RESULTS: Here we describe a 3 months old male child with normal pre- and postnatal development and with a de novo ring supernumerary marker chromosome in amniocytes cultures. Using new fluorescence in situ hybridization (FISH) techniques, three distinguishable sSMCs (cryptic mosaicism), all derived from chromosome 20, were observed, including ring and minute chromosomes. This heterogeneity was impossible to detect by the conventional G-banding technique or conventional FISH technique that were used before the application of new FISH techniques (subcentromere-specific multicolor FISH [subcenM-FISH]) and a probe, specific for the 20p12.2 band. The sSMC present in 25% of the cells was present as r(20)(::p12.2~12.3->q11.1::)5/r(20;20)(::p12.1 >q11.1::q11.1 >p12.1::)2/min(20;20)(:p12.1->q11.1::q11.1->p12.1:)1. The final karyotype was 47,XY,+r(20)[25%]/46,XY[75%]. CONCLUSION: We emphasize the importance of application of molecular cytogenetics in a prenatally diagnostic laboratory and description of more cases to enable a better genetic counseling and risk evaluation. PMID- 19128451 TI - Metabolic engineering of a reduced-genome strain of Escherichia coli for L threonine production. AB - BACKGROUND: Deletion of large blocks of nonessential genes that are not needed for metabolic pathways of interest can reduce the production of unwanted by products, increase genome stability, and streamline metabolism without physiological compromise. Researchers have recently constructed a reduced-genome Escherichia coli strain MDS42 that lacks 14.3% of its chromosome. RESULTS: Here we describe the reengineering of the MDS42 genome to increase the production of the essential amino acid L-threonine. To this end, we over-expressed a feedback resistant threonine operon (thrA*BC), deleted the genes that encode threonine dehydrogenase (tdh) and threonine transporters (tdcC and sstT), and introduced a mutant threonine exporter (rhtA23) in MDS42. The resulting strain, MDS-205, shows an ~83% increase in L-threonine production when cells are grown by flask fermentation, compared to a wild-type E. coli strain MG1655 engineered with the same threonine-specific modifications described above. And transcriptional analysis revealed the effect of the deletion of non-essential genes on the central metabolism and threonine pathways in MDS-205. CONCLUSION: This result demonstrates that the elimination of genes unnecessary for cell growth can increase the productivity of an industrial strain, most likely by reducing the metabolic burden and improving the metabolic efficiency of cells. PMID- 19128452 TI - The constancy of gene conservation across divergent bacterial orders. AB - BACKGROUND: Orthologous genes are frequently presumed to perform similar functions. However, outside of model organisms, this is rarely tested. One means of inferring changes in function is if there are changes in the level of gene conservation and selective constraint. Here we compare levels of gene conservation across three bacterial groups to test for changes in gene functionality. FINDINGS: The level of gene conservation for different orthologous genes is highly correlated across clades, even for highly divergent groups of bacteria. These correlations do not arise from broad differences in gene functionality (e.g. informational genes vs. metabolic genes), but instead seem to result from very specific differences in gene function. Furthermore, these functional differences appear to be maintained over very long periods of time. CONCLUSION: These results suggest that even over broad time scales, most bacterial genes are under a nearly constant level of purifying selection, and that bacterial evolution is thus dominated by selective and functional stasis. PMID- 19128454 TI - Congenital bronchial atresia presenting as a cavitary lesion on chest radiography: a case report. AB - BACKGROUND: Congenital bronchial atresia is a rare anomaly, which usually presents in adulthood as an incidental finding on routine examinations. CASE PRESENTATION: In this report we present a patient with a cavitary lesion at his right upper lobe, found by chance on chest radiography. Computed tomography of the chest revealed the characteristic findings of a mucocele with distal oligemia and hyperlucency of the affected lung parenchyma. Further examination including bronchoscopy virtually excluded other possible disorders and the diagnosis of congenital bronchial atresia was established. CONCLUSION: The radiological presentation of congenital bronchial atresia may occasionally mimic serious lung diseases. The procedure of choice for the diagnosis is the computed tomography of the chest. Bronchoscopy is not diagnostic but is valuable in doubtful cases to exclude different disorders. PMID- 19128453 TI - Age patterns of severe paediatric malaria and their relationship to Plasmodium falciparum transmission intensity. AB - BACKGROUND: The understanding of the epidemiology of severe malaria in African children remains incomplete across the spectrum of Plasmodium falciparum transmission intensities through which communities might expect to transition, as intervention coverage expands. METHODS: Paediatric admission data were assembled from 13 hospitals serving 17 communities between 1990 and 2007. Estimates of Plasmodium falciparum transmission intensity in these communities were assembled to be spatially and temporally congruent to the clinical admission data. The analysis focused on the relationships between community derived parasite prevalence and the age and clinical presentation of paediatric malaria in children aged 0-9 years admitted to hospital. RESULTS: As transmission intensity declined a greater proportion of malaria admissions were in older children. There was a strong linear relationship between increasing transmission intensity and the proportion of paediatric malaria admissions that were infants (R2 = 0.73, p < 0.001). Cerebral malaria was reported among 4% and severe malaria anaemia among 17% of all malaria admissions. At higher transmission intensity cerebral malaria was a less common presentation compared to lower transmission sites. There was no obvious relationship between the proportions of children with severe malaria anaemia and transmission intensity. CONCLUSION: As the intensity of malaria transmission declines in Africa through the scaling up of insecticide-treated nets and other vector control measures a focus of disease prevention among very young children becomes less appropriate. The understanding of the relationship between parasite exposure and patterns of disease risk should be used to adapt malaria control strategies in different epidemiological settings. PMID- 19128455 TI - Artemisinin-based combinations versus amodiaquine plus sulphadoxine-pyrimethamine for the treatment of uncomplicated malaria in Faladje, Mali. AB - BACKGROUND: Because of the emergence of chloroquine resistance in Mali, artemether-lumefantrine (AL) or artesunate-amodiaquine (AS+AQ) are recommended as first-line therapy for uncomplicated malaria, but have not been available in Mali until recently because of high costs. METHODS: From July 2005 to January 2006, a randomized open-label trial of three oral antimalarial combinations, namely AS+AQ, artesunate plus sulphadoxine-pyrimethamine (AS+SP), and amodiaquine plus sulphadoxine-pyrimethamine (AQ+SP), was conducted in Faladje, Mali. Parasite genotyping by polymerase chain reaction (PCR) was used to distinguish new from recrudescent Plasmodium falciparum infections. RESULTS: 397 children 6 to 59 months of age with uncomplicated Plasmodium falciparum malaria were enrolled, and followed for 28 days to assess treatment efficacy. Baseline characteristics were similar in all three treatment groups. The uncorrected rates of adequate clinical and parasitologic response (ACPR) were 55.7%, 90.8%, and 97.7% in AS+AQ, AS+SP, and AQ+SP respectively (p < 0.001); after PCR correction ACPR rates were similar among treatment groups: 95.4%, 96.9%, and 99.2% respectively (p = 0.17). Mean haemoglobin concentration increased across all treatment groups from Day 0 (9.82 +/- 1.68 g/dL) to Day 28 (10.78 +/- 1.49 g/dL) (p < 0.001), with the greatest improvement occurring in children treated with AQ+SP. On Day 2, the prevalence of parasitaemia was significantly greater among children treated with AQ+SP (50.8%) than in children treated with AS+AQ (10.5%) or AS+SP (10.8%) (p < 0.001). No significant difference in gametocyte carriage was found between groups during the follow-up period. CONCLUSION: The combination of AQ+SP provides a potentially low cost alternative for treatment of uncomplicated P. falciparum infection in Mali and appears to have the added value of longer protective effect against new infection. PMID- 19128456 TI - Inhibition of HIF-1alpha activity by homeodomain-interacting protein kinase-2 correlates with sensitization of chemoresistant cells to undergo apoptosis. AB - BACKGROUND: Homeodomain-interacting protein kinase-2 (HIPK2), a transcriptional co-repressor with apoptotic function, can affect hypoxia-inducible factor 1 (HIF 1) transcriptional activity, through downmodulation of its HIF-1alpha subunit, in normoxic condition. Under hypoxia, a condition often found in solid tumors, HIF 1alpha is activated to induce target genes involved in chemoresistance, inhibition of apoptosis and tumor progression. Here, we investigated whether the HIPK2 overexpression could downregulate HIF-1alpha expression and activity in tumor cells treated with hypoxia-mimicking condition, and evaluated whether HIPK2 dependent downregulation of HIF-1alpha could sensitize chemoresistant tumor cells to adriamycin (ADR)-induced apoptosis. METHODS: Tumor cell lines carrying wild type p53, siRNA p53, or mutant p53 were overexpressed with HIPK2 (full length or catalytic inactive mutant) and treated with cobalt chloride (CoCl2) to mimic hypoxia, in the presence or absence of ADR treatment. HIF-1alpha expression was measured by semiquantitative reverse-transcriptase (RT)-PCR and Western immunoblotting and HIF-1 activity was evaluated by luciferase assay using reporter plasmid containing hypoxia response elements (HREs) upstream of luciferase gene. HIF-1 target genes, including multidrug resistance 1 (MDR1) and the antiapoptotic Bcl2 were determined by RT-PCR. Cell survival and apoptosis were measured by colony assay and cleavage of the caspase-3 substrate PARP, respectively. RESULTS: Overexpression of HIPK2 resulted in downmodulation of cobalt-stabilized HIF-1alpha protein and HIF-1alpha mRNA levels, with subsequent inhibition of HIF-1 transcriptional activity. MDR1 and Bcl-2 gene expression was downmodulated by HIPK2 overexpression in cobalt-treated cells. Inhibition of HIF 1 transcriptional activity was dependent on HIPK2 catalytic activity. HIPK2 overexpression did not induce per se apoptosis of cobalt-treated cells, on the contrary it sensitized cobalt-treated cells to ADR-induced apoptosis, regardless of their p53 status. CONCLUSION: The ability of HIPK2 to restore the apoptosis inducing potential of chemotherapeutic drug in hypoxia-mimicking condition and therefore to sensitize chemoresistant tumor cells suggests that HIPK2 may induce fundamental alterations in cell signaling pathways, involving or not p53 function. Thus potential use of HIPK2 is promising for cancer treatment by potentiating cytotoxic therapies, regardless of p53 cell status. PMID- 19128457 TI - European Practice Assessment of Cardiovascular risk management (EPA Cardio): protocol of an international observational study in primary care. AB - BACKGROUND: Despite important improvements in available prevention and treatment, cardiovascular diseases (CVD) remain an important cause of morbidity and mortality. Not all high-risk patients and patients with CVD have healthy lifestyles and receive the best possible healthcare. Internationally comparative data are needed to compare cardiovascular risk management in different countries, and to examine the impact of improvement programs and others factors. OBJECTIVES: This study aims to provide internationally comparative data on cardiovascular risk management provided in primary care and on health-related lifestyles of patients in Europe. The study will also explore the views of doctors and patients on innovative preventive services for CVDs. DESIGN AND METHODS: An observational cross-sectional study is planned. In 10 European countries, stratified samples of 36 practices per country will be recruited. In each practice, three samples of 15 patients each will be sampled: patients with coronary heart disease, patients at high risk for CVD, and healthy adult patients. The quality of cardiovascular risk management has been specified in terms of 44 performance indicators that resulted from an international Delphi-procedure with general practitioners. Most indicators are based on medical records, and some on a structured interview with a contact person of the practice. Lifestyle (smoking, physical exercise, diet) will be measured with previously validated questionnaires that are completed by patients. Additional measures include practice characteristics and exposure to programs to improve cardiovascular care. PMID- 19128458 TI - Independent mobility in relation to weekday and weekend physical activity in children aged 10-11 years: The PEACH Project. AB - BACKGROUND: Children's independent mobility has fallen in recent years and may in part explain reported declines in physical activity in young people. This cross sectional study investigated whether independent mobility in boys and girls was related to objectively measured physical activity. METHODS: Thirteen hundred and seven 10-11 year old boys and girls from 23 schools in a large UK city took part. Measures included objectively recorded physical activity (accelerometer (Actigraph GT1M)), height (m) and weight (kg), a newly developed scale for local (Local-IM) and area independent mobility (Area-IM), minutes of daylight after school, level of neighbourhood deprivation and pubertal status. RESULTS: Boys had greater Local-IM, Area-IM and physical activity (average weekday and weekend counts per minute) compared to girls. In linear regression analyses (adjusting for minutes of daylight after school, neighbourhood deprivation, pubertal status and body mass index) higher scores for Local-IM and Area-IM were significantly (p < 0.01) related to higher levels of physical activity on weekdays for boys and girls. For weekend physical activity, only Local-IM in girls remained significant (p < 0.05) in the model. CONCLUSION: Independent mobility appears to be an important independent correlate of weekday physical activity for both boys and girls. PMID- 19128459 TI - Reduced use of antimicrobials after vaccination of pigs against porcine proliferative enteropathy in a Danish SPF herd. AB - The present study explored whether the use of group medication with antibiotics in a Danish pig herd was reduced after vaccination of the pigs against proliferative enteropathy (PE) caused by Lawsonia intracellularis. 7900 pigs originating from a single commercial sow herd were vaccinated against L. intracellularis, whereas 7756 pigs were kept as non-vaccinated controls. The pigs were included batch-wise in the study with every second batch being vaccinated. In the vaccinated batches, the consumption of oxytetracykline to treat PE was reduced by 79%, with a significantly lower number of pigs being treated (P < 0.0001). Vaccination also resulted in a highly significant improvement of average daily weight gain (+ 46 g/day; P = 9.55 x 10(-31)) and carcase weight (+ 1.25 kg; P = 4.54 x 10(-05)) as well as a shortened fattening period (-8 days; P = 2.01 x 10(-45)). PMID- 19128460 TI - Development of hepatocellular carcinoma in a patient 13 years after sustained virological response to interferon against chronic hepatitis C: a case report. AB - BACKGROUND: Although several recent reports have shown that hepatocellular carcinoma (HCC) developed in patients with chronic hepatitis C (CH-C) even after having a sustained virological response (SVR) to interferon (IFN) therapy, it is not common for HCC to develop more than 10 years after SVR. CASE PRESENTATION: A 73-year-old Japanese man with CH-C who achieved SVR to IFN therapy 13 years ago was admitted into our hospital because of huge multiple liver tumors along with marked elevation of the tumor markers. Several diagnostic modalities strongly suggested HCC, and we performed histopathological examination. After confirming the diagnosis as well-differentiated HCC, we successfully treated these tumors with intensive combination therapies. CONCLUSION: Our report highlights the need for careful follow-up for more than 10 years even if the patients with CH-C achieve SVR to IFN therapy. PMID- 19128461 TI - Fauna used in popular medicine in Northeast Brazil. AB - BACKGROUND: Animal-based remedies constitute an integral part of Brazilian Traditional Medicine. Due to its long history, zootherapy has in fact become an integral part of folk medicine both in rural and urban areas of the country. In this paper we summarize current knowledge on zootherapeutic practices in Northeast of Brazil, based on information compiled from ethnobiological scientific literature. METHODS: In order to examine the diversity of animals used in traditional medicine in Northeast of Brazil, all available references or reports of folk remedies based on animals sources were examined. 34 sources were analyzed. Only taxa that could be identified to species level were included in assessment of medicinal animal species. Scientific names provided in publications were updated. RESULTS: The review revealed that at least 250 animal species (178 vertebrates and 72 invertebrates) are used for medicinal purposes in Northeast of Brazil. The inventoried species comprise 10 taxonomic categories and belong to 141 Families. The groups with the greatest number of species were fishes (n = 58), mammals (n = 47) and reptiles (n = 37). The zootherapeutical products are used for the treatment of different illnesses. The most widely treated condition were asthma, rheumatism and sore throat, conditions, which had a wide variety of animals to treat them with. Many animals were used for the treatment of multiple ailments. Beyond the use for treating human diseases, zootherapeutical resources are also used in ethnoveterinary medicine CONCLUSION: The number of medicinal species catalogued was quite expressive and demonstrate the importance of zootherapy as alternative therapeutic in Northeast of Brazil. Although widely diffused throughout Brazil, zootherapeutic practices remain virtually unstudied. There is an urgent need to examine the ecological, cultural, social, and public health implications associated with fauna usage, including a full inventory of the animal species used for medicinal purposes and the socio-cultural context associated with their consumption. PMID- 19128462 TI - Prevalence and inter-relationship of different Doppler measures of dyssynchrony in patients with heart failure and prolonged QRS: a report from CARE-HF. AB - BACKGROUND: Cardiac resynchronisation therapy (CRT) improves mortality and morbidity in heart failure patients with wide QRS. Observational studies suggest that patients having more left ventricular dyssynchrony pre-implantation obtain greater benefit on ventricular function and symptoms with CRT. AIM: To provide an analysis of the prevalence and type of dyssynchrony in patients included in the CARE-HF trial. METHODS: 100 patients 67 (58 to 71) years were examined with echocardiography including tissue doppler imaging before receiving a CRT pacemaker. Atrio-ventricular dyssynchrony (LVFT/RR) was defined as left ventricular filling time <40% of the RR-interval. Inter-ventricular mechanical delay (IVMD) was measured as the difference in onset of Doppler-flow in the pulmonary and aortic outflow tracts >40 ms. Intra-ventricular (regional) dyssynchrony in a 16-segment model was expressed either as a delayed longitudinal contraction (DLC) during the postsystolic phase or by tissue synchronisation imaging (TSI) with a predefined time-difference in systolic maximal velocities >85 ms. RESULTS: LVFT/RR was present in 34% and IVMD in 60% of patients while intra-ventricular dyssynchrony was present in 85% (DLC) and 86% (TSI) with a high agreement between the measures (Kappascore 0.86-1.00), indicating the methods being interchangeable. Patients with cardiomyopathy (53%) were more likely to have LVFT/RR <40% (45% vs. 21% (p= 0.02)) and more segments affected by intra ventricular dyssynchrony 4(3, 5) vs. 3(1, 4), p = 0.002, compared to patients with ischemic heart disease. CONCLUSION: The prevalence of intra-ventricular dyssynchrony is high in patients with heart failure, wide QRS and depressed systolic function. Most important, TSI appears to be a fast and reliable method to identify patients with intra-ventricular dyssynchrony likely to benefit from CRT. PMID- 19128463 TI - Intraductal and invasive adenocarcinoma of duct of Luschka, mimicking chronic cholecystitis and cholelithiasis. AB - BACKGROUND: Intraductal and invasive adenocarcinoma of duct of Luschka is rare. To the best of our knowledge, this is the second case report of intraductal and invasive carcinoma arising from ducts of Luschka. CASE PRESENTATION: Patient presented to hospital with signs and symptoms of chronic cholecystitis and cholelithiasis. Ultrasound examination revealed thickening of gallbladder wall with abnormal septation around liver bed. Patient underwent laparoscopic cholecystectomy and resection of the adjacent liver bed. Histologic examination confirmed an intraductal and invasive adenocarcinoma arising from Luschka ducts. CONCLUSION: Adenocarcinoma of ducts of Luschka should be considered among differential diagnoses for the patients with typical clinical presentations of chronic cholecystitis and cholelithiasis. PMID- 19128464 TI - Effects of probiotic bacteria, isoflavones and simvastatin on lipid profile and atherosclerosis in cholesterol-fed rabbits: a randomized double-blind study. AB - BACKGROUND: Much attention has been drawn to different alternative strategies for cardiovascular disease prevention. OBJECTIVE: The aim of the present study was to observe and compare the effects of Enterococcus faecium CRL183 (probiotic microorganism), an isoflavones mixture and simvastatin (drug used to treat hypercholesterolemia) on lipid parameters and atherosclerosis development in rabbits with induced hypercholesterolemia. METHODS: The animals were randomly allocated to 5 experimental groups (n = 6) for 60 days: control (C) that did not consume cholesterol, hypercholesterolemic (H) that consumed an atherogenic diet (1.0% cholesterol wt/wt), hypercholesterolemic plus E. faecium (HE), hypercholesterolemic plus isoflavone (HI) and hypercholesterolemic plus simvastatin (HS). Total and HDL-cholesterol and triglycerides were determined by enzymatic methods; non-HDL-C was calculated by subtracting HDL-C from total cholesterol; and atherosclerosis was presented as the percentage of lesion area, relative to the total area from the aorta segment analyzed. RESULTS: Simvastatin significantly reduced the tot cholesterol (16%) and non-HDL-C level (17%) and increased the HDL-C (98%), compared to group H. E. faecium raised the HDL-C level by 43.3% (P < 0.05). Isoflavone decreased the total cholesterol and non-HDL-C concentrations (9%), but this effect was not statistically significant. At the end of the treatments, groups HE and HS had significantly lower levels of triglycerides in relation to H and HI. The atherosclerotic lesion area in the aortic arch was not different between groups. The extent of atherosclerosis in the thoracic and abdominal aorta was reduced in the groups HI and HS by 22.7% and 26.7% respectively, but this effect was not significant (P > 0.05). CONCLUSION: The results indicated that probiotic microorganism E. faecium CRL 183 could be used to improve the lipid profile as an alternative or an adjuvant for drug therapy. The effectiveness of simvastatin in the management of blood lipid was confirmed. There were no effects of soy isoflavones, E. faecium and simvastatin on atherosclerosis development. PMID- 19128465 TI - Evaluation of gene amplification and protein expression of HER-2/neu in esophageal squamous cell carcinoma using Fluorescence in situ Hybridization (FISH) and immunohistochemistry. AB - BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is the sixth most frequent neoplasia in Brazil. It is usually associated with a poor prognosis because it is often at an advanced stage when diagnosed and there is a high frequency of lymph node metastases. It is important to know what prognostic factors can facilitate diagnosis, optimize therapeutic decisions, and improve the survival of these patients. A member of the epidermal growth factor receptor (EGFR) family, c-erbB 2, has received much attention because of its therapeutic implications; however, few studies involving fluorescence in situ hybridization (FISH) analysis of HER 2/neu gene amplification and protein expression in ESCC have been conducted. The aim of this study was to verify the presence of HER-2/neu gene amplification using FISH, and to correlate the results with immunohistochemical expression and clinical-pathological findings. METHODS: One hundred and ninety-nine ESCC cases were evaluated using the Tissue Microarray (TMA) technique. A polyclonal antibody against c-erbB-2 was used for immunohistochemistry. Analyses were based on the membrane staining pattern. The results were classified according to the Herceptest criteria (DAKO): negative (0/1+), potential positive (2+) and positive (3+). The FISH reactions were performed according to the FISH HER2 PharmDx (DAKO) protocol. In each case, 100 tumor nuclei were evaluated. Cases showing a gene/CEN17 fluorescence ratio > or = 2 were considered positive for gene amplification. RESULTS: The c-erbB-2 expression was negative in 117/185 cases (63.2%) and positive in 68 (36.8%), of which 56 (30.3%) were 2+ and 12 (6.5%) were 3+. No significant associations were found among protein expression, clinicopathological data and overall survival. Among the 47 cases analyzed, 38 (80.9%) showed no gene amplification while 9 (19.1%) showed amplification, as demonstrated by FISH. Cases that were negative (0/1+) and potential positive (2+) for c-erbB-2 expression by immunohistochemistry showed no gene amplification. However, all cases with gene amplification were positive (3+) by immunohistochemistry. According to univariate analysis, there was a significant difference (p = 0.003) in survival rates when cases with and without HER-2/neu amplification were compared. CONCLUSION: Our data demonstrate the correspondence between gene amplification and protein expression of HER-2/neu. Gene amplification is an indicator of poor prognosis in ESCC. PMID- 19128466 TI - Reporter gene-expressing bone marrow-derived stromal cells are immune-tolerated following implantation in the central nervous system of syngeneic immunocompetent mice. AB - BACKGROUND: Cell transplantation is likely to become an important therapeutic tool for the treatment of various traumatic and ischemic injuries to the central nervous system (CNS). However, in many pre-clinical cell therapy studies, reporter gene-assisted imaging of cellular implants in the CNS and potential reporter gene and/or cell-based immunogenicity, still remain challenging research topics. RESULTS: In this study, we performed cell implantation experiments in the CNS of immunocompetent mice using autologous (syngeneic) luciferase-expressing bone marrow-derived stromal cells (BMSC-Luc) cultured from ROSA26-L-S-L Luciferase transgenic mice, and BMSC-Luc genetically modified using a lentivirus encoding the enhanced green fluorescence protein (eGFP) and the puromycin resistance gene (Pac) (BMSC-Luc/eGFP/Pac). Both reporter gene-modified BMSC populations displayed high engraftment capacity in the CNS of immunocompetent mice, despite potential immunogenicity of introduced reporter proteins, as demonstrated by real-time bioluminescence imaging (BLI) and histological analysis at different time-points post-implantation. In contrast, both BMSC-Luc and BMSC Luc/eGFP/Pac did not survive upon intramuscular cell implantation, as demonstrated by real-time BLI at different time-points post-implantation. In addition, ELISPOT analysis demonstrated the induction of IFN-gamma-producing CD8+ T-cells upon intramuscular cell implantation, but not upon intracerebral cell implantation, indicating that BMSC-Luc and BMSC-Luc/eGFP/Pac are immune-tolerated in the CNS. However, in our experimental transplantation model, results also indicated that reporter gene-specific immune-reactive T-cell responses were not the main contributors to the immunological rejection of BMSC-Luc or BMSC Luc/eGFP/Pac upon intramuscular cell implantation. CONCLUSION: We here demonstrate that reporter gene-modified BMSC derived from ROSA26-L-S-L-Luciferase transgenic mice are immune-tolerated upon implantation in the CNS of syngeneic immunocompetent mice, providing a research model for studying survival and localisation of autologous BMSC implants in the CNS by real-time BLI and/or histological analysis in the absence of immunosuppressive therapy. PMID- 19128467 TI - Evaluation of bioluminescent imaging for noninvasive monitoring of colorectal cancer progression in the liver and its response to immunogene therapy. AB - BACKGROUND: Bioluminescent imaging (BLI) is based on the detection of light emitted by living cells expressing a luciferase gene. Stable transfection of luciferase in cancer cells and their inoculation into permissive animals allows the noninvasive monitorization of tumor progression inside internal organs. We have applied this technology for the development of a murine model of colorectal cancer involving the liver, with the aim of improving the pre-clinical evaluation of new anticancer therapies. RESULTS: A murine colon cancer cell line stably transfected with the luciferase gene (MC38Luc1) retains tumorigenicity in immunocompetent C57BL/6 animals. Intrahepatic inoculation of MC38Luc1 causes progressive liver infiltration that can be monitored by BLI. Compared with ultrasonography (US), BLI is more sensitive, but accurate estimation of tumor mass is impaired in advanced stages. We applied BLI to evaluate the efficacy of an immunogene therapy approach based on the liver-specific expression of the proinflammatory cytokine interleukin-12 (IL-12). Individualized quantification of light emission was able to determine the extent and duration of antitumor responses and to predict long-term disease-free survival. CONCLUSION: We show that BLI is a rapid, convenient and safe technique for the individual monitorization of tumor progression in the liver. Evaluation of experimental treatments with complex mechanisms of action such as immunotherapy is possible using this technology. PMID- 19128468 TI - Supplementary feeding affects the breeding behaviour of male European treefrogs (Hyla arborea). AB - BACKGROUND: We investigated the effects of energetic constraints on the breeding behaviour of male European treefrogs Hyla arborea and how calling males allocated additional energy supplied by feeding experiments. RESULTS: Presence in the chorus was energetically costly indicated by both fed and unfed males losing weight. Males that were supplied with additional energy did not show longer chorus tenure. Instead, fed males returned sooner to the chorus. Additionally, fed males called more often than control males, a novel response for anurans. A significantly higher calling rate was noted from males even 31 nights after supplementary feeding. CONCLUSION: This strategy of allocating additional energy reserves to increasing calling rate is beneficial given the preference of female hylids for males calling at high rates and a female's ability to detect small incremental increases in calling rate. PMID- 19128469 TI - Losing the genetic twin: donor grief after unsuccessful unrelated stem cell transplantation. AB - BACKGROUND: Stem cell transplantations from related or unrelated donors are used to cure leukaemia and other blood diseases. When a patient dies after an unsuccessful transplantation, interested unrelated donors are informed about the failure by their donor centre. Studies focussing on failed related donations show that donors undergo an intense grieving process. As there are only two investigations about reactions from unrelated donors, knowledge about their reactions is less comprehensive. METHODS: We conducted a prospective study of reactions of unrelated donors to the information of failed transplantations, subject to various communication methods (letter, phone). Questionnaires were sent to 395 unrelated donors who received the news of their recipients' deaths between November 2005 and August 2006. In addition, twelve in-depth interviews with selected donors were carried out. RESULTS: Unrelated donors were emotionally affected by the recipients' deaths, and it is appropriate to speak about a "Donor Grief" phenomenon, as the results of 325 returned questionnaires (return rate 82.3%) and in-depth interviews show. Donors demonstrated a range of feelings such as sadness, disappointment, grief, and helplessness. These feelings were often unexpectedly intense given the fact that the recipient was a stranger. Although the news caused grief, donors underlined that they nevertheless wanted to be informed. They preferred knowledge of the failure to uncertainty. The method of providing the information is only of secondary importance. Most donors favoured the way of communication they had experienced. CONCLUSION: This result indicates that both phone and letter communication can be justified. However, phone communication seems to be superior with respect to aspects of sensitivity. In spite of transplantation failure and the associated negative feelings, most donors were happy to have donated and would be willing to do so again. Our results underline the special responsibility of donor centres for informing and supporting unrelated volunteer donors in case their recipients have died. PMID- 19128470 TI - Impaired IGF1-GH axis and new therapeutic options in Alstrom Syndrome patients: a case series. AB - BACKGROUND: Defects of the primary cilium and its anchoring structure, the basal body, cause a number of human genetic disorders, collectively termed ciliopathies: primary ciliary dyskinesia, Bardet-Biedl syndrome, polycystic kidney and liver disease, nephronophthisis, Alstrom syndrome, Meckel-Gruber syndrome and some forms of retinal degeneration.Alstrom syndrome is an extremely rare, autosomal recessive genetic disorder characterized by a group of signs and symptoms including infantile onset dilated cardiomyopathy, blindness, hearing impairment/loss, obesity, diabetes, hepatic and renal dysfunction.Because adult growth hormone deficiency and Alstrom Syndrome share some clinical and metabolic features, we studied the GH-IGF1 axis, using MRI techniques and dynamic tests in 3 unrelated patients with Alstrom syndrome. CASE PRESENTATION: The patients were hospitalized and the growth hormone stimulatory tests were made, as well as brain MRI. Insulin provocative test revealed a severe GH deficiency in these patients, defined by a peak response to insulin-induced hypoglycemia less than 3 ng/dl and IGF1 concentrations less than - 2SDS.We didn't find multiple pituitary hormone deficiency and we noticed only a severe GH deficiency in all three patients. The MRI study of the diencephalic and pituitary region was suggestive for the diagnosis of empty sella in one patient.One patient received Recombinant-GH replacement for one year with very good results, one underwent a gastric sleeve with a satisfactory outcome, one patient died due to the progression of the cardiac myopathy. CONCLUSION: Future studies are needed to assses if the substitution therapy with Recombinant Growth hormone is cost-effective and without risk in such patients with Alstrom Syndrome and severe insulin resistance, despite our good results in one patient. Also, careful clinical and genetic studies can contribute to a better understanding of the evolution after different therapeutical attempt in the complex disorders such as Alstrom Syndrome. PMID- 19128471 TI - Atypical right diaphragmatic hernia (hernia of Morgagni), spigelian hernia and epigastric hernia in a patient with Williams syndrome: a case report. AB - INTRODUCTION: Williams syndrome is rare genetic disorder resulting in neurodevelopmental problems. Hernias of the foramen of Morgagni are rare diaphragmatic hernias and they mostly present on the right side, in the anterior mediastinum. They are usually asymptomatic and are difficult to diagnose, especially in patients with learning disabilities. CASE PRESENTATION: This 49 year-old woman with Williams syndrome, cognitive impairment and aortic stenosis presented to physicians with right-sided chest pain. She had previously undergone repair of her right spigelian and epigastric hernia. Her abdominal examination was unremarkable. Chest X-ray suggested right-sided diaphragmatic hernia and pleural effusion for which she received treatment. The computed tomography scan showed a diaphragmatic hernia with some collapse/consolidation of the adjacent lung. Furthermore, the patient had aortic stenosis and was high risk for anaesthesia (ASA grade 3). She underwent successful laparoscopic repair of her congenital diaphragmatic hernia leading to a quick and uneventful postoperative recovery. CONCLUSION: These multiple hernias suggest that patients with Williams syndrome may have some connective tissue disorder which makes them prone to develop hernias especially associated with those parts of the body which may have intracavity pressure variations like the abdomen. Diaphragmatic hernia may be the cause of chest pain in these patients. A computed tomography scan helps in early diagnosis, and laparoscopic repair helps in prevention of further complications, and leads to quick recovery especially in patients with learning disabilities. In the presence of significant comorbidities, a less invasive operative procedure with quick recovery becomes advisable. PMID- 19128472 TI - ContDist: a tool for the analysis of quantitative gene and promoter properties. AB - BACKGROUND: The understanding of how promoter regions regulate gene expression is complicated and far from being fully understood. It is known that histones' regulation of DNA compactness, DNA methylation, transcription factor binding sites and CpG islands play a role in the transcriptional regulation of a gene. Many high-throughput techniques exist nowadays which permit the detection of epigenetic marks and regulatory elements in the promoter regions of thousands of genes. However, so far the subsequent analysis of such experiments (e.g. the resulting gene lists) have been hampered by the fact that currently no tool exists for a detailed analysis of the promoter regions. RESULTS: We present ContDist, a tool to statistically analyze quantitative gene and promoter properties. The software includes approximately 200 quantitative features of gene and promoter regions for 7 commonly studied species. In contrast to "traditionally" ontological analysis which only works on qualitative data, all the features in the underlying annotation database are quantitative gene and promoter properties.Utilizing the strong focus on the promoter region of this tool, we show its usefulness in two case studies; the first on differentially methylated promoters and the second on the fundamental differences between housekeeping and tissue specific genes. The two case studies allow both the confirmation of recent findings as well as revealing previously unreported biological relations. CONCLUSION: ContDist is a new tool with two important properties: 1) it has a strong focus on the promoter region which is usually disregarded by virtually all ontology tools and 2) it uses quantitative (continuously distributed) features of the genes and its promoter regions which are not available in any other tool. ContDist is available from http://web.bioinformatics.cicbiogune.es/CD/ContDistribution.php. PMID- 19128473 TI - Mesothelin, Stereocilin, and Otoancorin are predicted to have superhelical structures with ARM-type repeats. AB - BACKGROUND: Mesothelin is a 40 kDa protein present on the surface of normal mesothelial cells and overexpressed in many human tumours, including mesothelioma and ovarian and pancreatic adenocarcinoma. It forms a strong and specific complex with MUC16, which is also highly expressed on the surface of mesothelioma and ovarian cancer cells. This binding has been suggested to be the basis of ovarian cancer metastasis. Knowledge of the structure of this protein will be useful, for example, in building a structural model of the MUC16-mesothelin complex. Mesothelin is produced as a precursor, which is cleaved by furin to produce the N terminal half, which is called the megakaryocyte potentiating factor (MPF), and the C-terminal half, which is mesothelin. Little is known about the function of mesothelin and there is no information on its possible three-dimensional structure. Mesothelin has been reported to be homologous to the deafness-related inner ear proteins otoancorin and stereocilin, for neither of which the three dimensional structure is known. RESULTS: The BLAST and PSI-BLAST searches confirmed that mesothelin and mesothelin precursor proteins are remotely homologous to stereocilin and otoancorin and more closely homologous to the hypothetical protein MPFL (MPF-like). Secondary structure prediction servers predicted a predominantly helical structure for both mesothelin and mesothelin precursor proteins and also for stereocilin and otoancorin. Three-dimensional structure prediction servers INHUB and I-TASSER produced structural models for mesothelin, which consisted of superhelical structures with ARM-type repeats in conformity with the secondary structure predictions. Similar ARM-type superhelical repeat structures were predicted by 3D-PSSM server for mesothelin precursor and for stereocilin and otoancorin proteins. CONCLUSION: The mesothelin superfamily of proteins, which includes mesothelin, mesothelin precursor, megakaryocyte potentiating factor, MPFL, stereocilin and otoancorin, are predicted to have superhelical structures with ARM-type repeats. We suggest that all of these function as superhelical lectins to bind the carbohydrate moieties of extracellular glycoproteins. PMID- 19128475 TI - No short-cut in assessing trial quality: a case study. AB - BACKGROUND: Assessing the quality of included trials is a central part of a systematic review. Many check-list type of instruments for doing this exist. Using a trial of antibiotic treatment for acute otitis media, Burke et al., BMJ, 1991, as the case study, this paper illustrates some limitations of the check list approach to trial quality assessment. RESULTS: The general verdict from the check list type evaluations in nine relevant systematic reviews was that Burke et al. (1991) is a good quality trial. All relevant meta-analyses extensively used its data to formulate therapeutic evidence. My comprehensive evaluation, on the other hand, brought to the surface a series of serious problems in the design, conduct, analysis and report of this trial that were missed by the earlier evaluations. CONCLUSION: A check-list or instrument based approach, if used as a short-cut, may at times rate deeply flawed trials as good quality trials. Check lists are crucial but they need to be augmented with an in-depth review, and where possible, a scrutiny of the protocol, trial records, and original data. The extent and severity of the problems I uncovered for this particular trial warrant an independent audit before it is included in a systematic review. PMID- 19128474 TI - The iron/heme regulated genes of Haemophilus influenzae: comparative transcriptional profiling as a tool to define the species core modulon. AB - BACKGROUND: Haemophilus influenzae requires heme for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. Although an understanding of the heme acquisition mechanisms of H. influenzae is emerging, significant gaps in our knowledge remain. Unresolved issues include the identities of all genes exhibiting altered transcription in response to iron and heme availability, the fraction of such genes functioning in iron/heme acquisition, and the heterogeneity of this gene set among clinical isolates. Previously we utilized H. influenzae strain Rd KW20 to demonstrate the utility of transcriptional profiling in defining the genes exhibiting altered transcription in response to environmental iron and heme levels. The current study expands upon those observations by determining the iron/heme modulons of two clinical isolates, the type b isolate 10810 and the nontypeable isolate R2866. These data are used to begin to define the core iron/heme modulon of the species. RESULTS: Microarray studies were performed to compare gene expression on transition from iron/heme restricted to iron/heme-replete conditions for each isolate. Of 1820 ORFs on the array corresponding to R2866 genes, 363 were significantly differentially expressed: 233 were maximally transcribed under iron/heme-replete conditions and 130 under iron/heme-restricted conditions. Of the 1883 ORFs representing genes of strain 10810, 353 were significantly differentially transcribed: 150 were preferentially transcribed under iron/heme-replete conditions and 203 under iron/heme-restricted conditions. Comparison of the data sets indicated that 163 genes exhibited similar regulation in both isolates and that 74 of these exhibited similar patterns of regulation in Rd KW20. These comprise the putative core iron/heme modulon. CONCLUSION: This study provides evidence for a conserved core of H. influenzae genes the transcription of which is altered by the availability of iron and/or heme in the growth environment. Elucidation of this modulon provides a means to identify genes with unrecognized roles in iron/heme acquisition or homeostasis, unanticipated responsiveness to environmental levels of the micronutrients or potential roles in virulence. Defining these core genes is also of potential importance in identifying targets for therapeutic and vaccine designs since products of these genes are likely to be preferentially expressed during growth in iron/heme restricted sites of the human body. PMID- 19128476 TI - Upstream sequence elements direct post-transcriptional regulation of gene expression under stress conditions in yeast. AB - BACKGROUND: The control of gene expression in eukaryotic cells occurs both transcriptionally and post-transcriptionally. Although many genes are now known to be regulated at the translational level, in general, the mechanisms are poorly understood. We have previously presented polysomal gradient and array-based evidence that translational control is widespread in a significant number of genes when yeast cells are exposed to a range of stresses. Here we have re examined these gene sets, considering the role of UTR sequences in the translational responses of these genes using recent large-scale datasets which define 5' and 3' transcriptional ends for many yeast genes. In particular, we highlight the potential role of 5' UTRs and upstream open reading frames (uORFs). RESULTS: We show a highly significant enrichment in specific GO functional classes for genes that are translationally up- and down-regulated under given stresses (e.g. carbohydrate metabolism is up-regulated under amino acid starvation). Cross-referencing these data with the stress response data we show that translationally upregulated genes have longer 5' UTRs, consistent with their role in translational regulation. In the first genome-wide study of uORFs in a set of mapped 5' UTRs, we show that uORFs are rare, being statistically under represented in UTR sequences. However, they have distinct compositional biases consistent with their putative role in translational control and are more common in genes which are apparently translationally up-regulated. CONCLUSION: These results demonstrate a central regulatory role for UTR sequences, and 5' UTRs in particular, highlighting the significant role of uORFs in post-transcriptional control in yeast. Yeast uORFs are more highly conserved than has been suggested, lending further weight to their significance as functional elements involved in gene regulation. It also suggests a more complex and novel mechanism of control, whereby uORFs permit genes to escape from a more general attenuation of translation under conditions of stress. However, since uORFs are relatively rare (only ~13% of yeast genes have them) there remain many unanswered questions as to how UTR elements can direct translational control of many hundreds of genes under stress. PMID- 19128477 TI - Auto-validating von Neumann rejection sampling from small phylogenetic tree spaces. AB - BACKGROUND: In phylogenetic inference one is interested in obtaining samples from the posterior distribution over the tree space on the basis of some observed DNA sequence data. One of the simplest sampling methods is the rejection sampler due to von Neumann. Here we introduce an auto-validating version of the rejection sampler, via interval analysis, to rigorously draw samples from posterior distributions over small phylogenetic tree spaces. RESULTS: The posterior samples from the auto-validating sampler are used to rigorously (i) estimate posterior probabilities for different rooted topologies based on mitochondrial DNA from human, chimpanzee and gorilla, (ii) conduct a non-parametric test of rate variation between protein-coding and tRNA-coding sites from three primates and (iii) obtain a posterior estimate of the human-neanderthal divergence time. CONCLUSION: This solves the open problem of rigorously drawing independent and identically distributed samples from the posterior distribution over rooted and unrooted small tree spaces (3 or 4 taxa) based on any multiply-aligned sequence data. PMID- 19128478 TI - Complex nature of SNP genotype effects on gene expression in primary human leucocytes. AB - BACKGROUND: Genome wide association studies have been hugely successful in identifying disease risk variants, yet most variants do not lead to coding changes and how variants influence biological function is usually unknown. METHODS: We correlated gene expression and genetic variation in untouched primary leucocytes (n = 110) from individuals with celiac disease - a common condition with multiple risk variants identified. We compared our observations with an EBV transformed HapMap B cell line dataset (n = 90), and performed a meta-analysis to increase power to detect non-tissue specific effects. RESULTS: In celiac peripheral blood, 2,315 SNP variants influenced gene expression at 765 different transcripts (< 250 kb from SNP, at FDR = 0.05, cis expression quantitative trait loci, eQTLs). 135 of the detected SNP-probe effects (reflecting 51 unique probes) were also detected in a HapMap B cell line published dataset, all with effects in the same allelic direction. Overall gene expression differences within the two datasets predominantly explain the limited overlap in observed cis-eQTLs. Celiac associated risk variants from two regions, containing genes IL18RAP and CCR3, showed significant cis genotype-expression correlations in the peripheral blood but not in the B cell line datasets. We identified 14 genes where a SNP affected the expression of different probes within the same gene, but in opposite allelic directions. By incorporating genetic variation in co-expression analyses, functional relationships between genes can be more significantly detected. CONCLUSION: In conclusion, the complex nature of genotypic effects in human populations makes the use of a relevant tissue, large datasets, and analysis of different exons essential to enable the identification of the function for many genetic risk variants in common diseases. PMID- 19128479 TI - Integrated taxonomy: traditional approach and DNA barcoding for the identification of filarioid worms and related parasites (Nematoda). AB - BACKGROUND: We compared here the suitability and efficacy of traditional morphological approach and DNA barcoding to distinguish filarioid nematodes species (Nematoda, Spirurida). A reliable and rapid taxonomic identification of these parasites is the basis for a correct diagnosis of important and widespread parasitic diseases. The performance of DNA barcoding with different parameters was compared measuring the strength of correlation between morphological and molecular identification approaches. Molecular distance estimation was performed with two different mitochondrial markers (coxI and 12S rDNA) and different combinations of data handling were compared in order to provide a stronger tool for easy identification of filarioid worms. RESULTS: DNA barcoding and morphology based identification of filarioid nematodes revealed high coherence. Despite both coxI and 12S rDNA allow to reach high-quality performances, only coxI revealed to be manageable. Both alignment algorithm, gaps treatment, and the criteria used to define the threshold value were found to affect the performance of DNA barcoding with 12S rDNA marker. Using coxI and a defined level of nucleotide divergence to delimit species boundaries, DNA barcoding can also be used to infer potential new species. CONCLUSION: An integrated approach allows to reach a higher discrimination power. The results clearly show where DNA-based and morphological identifications are consistent, and where they are not. The coherence between DNA based and morphological identification for almost all the species examined in our work is very strong. We propose DNA barcoding as a reliable, consistent, and democratic tool for species discrimination in routine identification of parasitic nematodes. PMID- 19128480 TI - Molecular evolution of UCP1 and the evolutionary history of mammalian non shivering thermogenesis. AB - BACKGROUND: Uncoupling protein 1 (UCP1) is a mitochondrial anion carrier, expressed in brown adipose tissue (BAT) of Eutherians. UCP1 is responsible for uncoupling mitochondrial proton transport from the production of ATP, thereby dissipating heat; it is essential for non-shivering thermogenesis (NST) in mammalian BAT. UCP1 orthologs have been identified in non-Eutherian mammals, fish and amphibians. Yet, UCP1 has a unique function in Eutherians in that it is necessary in the production of heat (NST). As such, this study aims to determine the evolutionary mode of UCP1 in Eutherians, where there is clear evidence of UCP1-dependent NST in BAT. RESULTS: Models of adaptive evolution through phylogenetic analysis of amino acid sequences by maximum likelihood were implemented to determine the mode of UCP1 protein evolution in Eutherians. An increase in the rate of amino acid substitutions on the branch leading to Eutherians is observed, but is best explained by relaxed constraints, not positive selection. Further, evidence for branch and site heterogeneity in selection pressures, as well as divergent selection pressures between UCP1 and its paralogs (UCP2-3) is observed. CONCLUSION: We propose that the unique thermogenic function of UCP1 in Eutherians may be best explained by neutral processes. Along with other evidence, this suggests that the primary biochemical properties of UCP1 may not differ between Eutherians and non-Eutherians. PMID- 19128481 TI - Schizophrenia genomics and proteomics: are we any closer to biomarker discovery? AB - The field of proteomics has made leaps and bounds in the last 10 years particularly in the fields of oncology and cardiovascular medicine. In comparison, neuroproteomics is still playing catch up mainly due to the relative complexity of neurological disorders. Schizophrenia is one such disorder, believed to be the results of multiple factors both genetic and environmental. Affecting over 2 million people in the US alone, it has become a major clinical and public health concern worldwide. This paper gives an update of schizophrenia biomarker research as reviewed by Lakhan in 2006 and gives us a rundown of the progress made during the last two years. Several studies demonstrate the potential of cerebrospinal fluid as a source of neuro-specific biomarkers. Genetic association studies are making headway in identifying candidate genes for schizophrenia. In addition, metabonomics, bioinformatics, and neuroimaging techniques are aiming to complete the picture by filling in knowledge gaps. International cooperation in the form of genomics and protein databases and brain banks is facilitating research efforts. While none of the recent developments described here in qualifies as biomarker discovery, many are likely to be stepping stones towards that goal. PMID- 19128482 TI - Readers as research detectives. AB - Flaws in research papers are common but it may require arduous detective work to unravel them. Checklists are helpful, but many inconsistencies will only be revealed through repeated cross-checks of every little detail, just like in a crime case. As a major deterrent for dishonesty, raw data from all trials should be posted on a public website. This would also make it much easier to detect errors and flaws in publications, and it would allow many research projects to be performed without collecting new data. The prevailing culture of secrecy and ownership to data is not in the best interests of patients. PMID- 19128483 TI - Identification of a rare de novo three-way complex t(5;20;8)(q31;p11.2;p21) with microdeletions on 5q31.2, 5q31.3, and 8p23.2 in a patient with hearing loss and global developmental delay: case report. AB - BACKGROUND: Complex chromosome rearrangements (CCRs), which involve more than two breakpoints on two or more chromosomes, are uncommon occurrences. Although most CCRs appear balanced at the level of the light microscope, many demonstrate cryptic, submicroscopic imbalances at the translocation breakpoints. RESULTS: We report a female with hearing loss and global developmental delay with a complex three-way unbalanced translocation (5;20;8)(q31;p11.2;p21) resulting in microdeletions on 5q31.2, 5q31.3, and 8p23.2 identified by karyotyping, microarray analysis and fluorescence in situ hybridization. DISCUSSION: The microdeletion of bands 8p23.2 may be associated with the hearing impairment. Furthermore, the characterization of this patient's chromosomal abnormalities demonstrates the importance of integrated technologies within contemporary cytogenetics laboratories. PMID- 19128484 TI - FORCA, a promoter element that responds to crosstalk between defense and light signaling. AB - BACKGROUND: Recognition of pathogenic microorganisms triggers in plants comprehensive transcriptional reprogramming. In order to identify transcriptome level control elements required for plant immune responses we are examining several sets of genes found by microarray experiments to be co-activated in Arabidopsis thaliana (Arabidopsis) seedlings infected with the oomycete Hyaloperonospora parasitica. Promoter motifs conserved in clusters of co expressed genes may be involved in mediating coordinated gene activity patterns. Although numerous studies identified such conserved promoter motifs in co expressed gene sets, reports confirming their function as regulatory elements are rare. RESULTS: FORCA is a hexameric promoter motif that is conserved in clusters of Arabidopsis genes co-expressed in response to fungal or oomycete pathogens as well as defined light treatments. FORCA is generally more frequently present in Arabidopsis promoter regions than statistically expected. It constitutively interacts in a DNA-sequence specific manner with nuclear Arabidopsis proteins. These interactions are suppressed by defense-related stimuli and enhanced by prolonged exposure to constant light. Furthermore FORCA mediates constitutive reporter gene expression in transiently transformed Nicotiana benthamiana leaves as well as in stably transformed Arabidopsis plants. Its responsiveness to defense-stimuli is modulated by the duration of light exposure. In plants grown under normal light conditions or constant darkness defense-related stimuli result in suppression of FORCA-mediated reporter gene expression, while in plants grown under constant light exposure, defense-induction results in enhanced FORCA mediated expression. In addition, we found plants subjected to constant light exposure to exhibit reduced susceptibility to virulent H. parasitica. CONCLUSION: We propose that FORCA is a regulatory cis-element that is present in a wide variety of Arabidopsis promoters. It integrates light- and defense-related signals and participates in adjusting the transcriptome to changes in environmental conditions. PMID- 19128485 TI - Catalytic inhibition of topoisomerase II by a novel rationally designed ATP competitive purine analogue. AB - BACKGROUND: Topoisomerase II poisons are in clinical use as anti-cancer therapy for decades and work by stabilizing the enzyme-induced DNA breaks. In contrast, catalytic inhibitors block the enzyme before DNA scission. Although several catalytic inhibitors of topoisomerase II have been described, preclinical concepts for exploiting their anti-proliferative activity based on molecular characteristics of the tumor cell have only recently started to emerge. Topoisomerase II is an ATPase and uses the energy derived from ATP hydrolysis to orchestrate the movement of the DNA double strands along the enzyme. Thus, interfering with ATPase function with low molecular weight inhibitors that target the nucleotide binding pocket should profoundly affect cells that are committed to undergo mitosis. RESULTS: Here we describe the discovery and characterization of a novel purine diamine analogue as a potent ATP-competitive catalytic inhibitor of topoisomerase II. Quinoline aminopurine compound 1 (QAP 1) inhibited topoisomerase II ATPase activity and decatenation reaction at sub-micromolar concentrations, targeted both topoisomerase II alpha and beta in cell free assays and, using a quantitative cell-based assay and a chromosome segregation assay, displayed catalytic enzyme inhibition in cells. In agreement with recent hypothesis, we show that BRCA1 mutant breast cancer cells have increased sensitivity to QAP 1. CONCLUSION: The results obtained with QAP 1 demonstrate that potent and selective catalytic inhibition of human topoisomerase II function with an ATP-competitive inhibitor is feasible. Our data suggest that further drug discovery efforts on ATP-competitive catalytic inhibitors are warranted and that such drugs could potentially be developed as anti-cancer therapy for tumors that bear the appropriate combination of molecular alterations. PMID- 19128486 TI - Production of recombinant AAV vectors encoding insulin-like growth factor I is enhanced by interaction among AAV rep regulatory sequences. AB - BACKGROUND: Adeno-associated virus (AAV) vectors are promising tools for gene therapy. Currently, their potential is limited by difficulties in producing high vector yields with which to generate transgene protein product. AAV vector production depends in part upon the replication (Rep) proteins required for viral replication. We tested the hypothesis that mutations in the start codon and upstream regulatory elements of Rep78/68 in AAV helper plasmids can regulate recombinant AAV (rAAV) vector production. We further tested whether the resulting rAAV vector preparation augments the production of the potentially therapeutic transgene, insulin-like growth factor I (IGF-I). RESULTS: We constructed a series of AAV helper plasmids containing different Rep78/68 start codon in combination with different gene regulatory sequences. rAAV vectors carrying the human IGF-I gene were prepared with these vectors and the vector preparations used to transduce HT1080 target cells. We found that the substitution of ATG by ACG in the Rep78/68 start codon in an AAV helper plasmid (pAAV-RC) eliminated Rep78/68 translation, rAAV and IGF-I production. Replacement of the heterologous sequence upstream of Rep78/68 in pAAV-RC with the AAV2 endogenous p5 promoter restored translational activity to the ACG mutant, and restored rAAV and IGF-I production. Insertion of the AAV2 p19 promoter sequence into pAAV-RC in front of the heterologous sequence also enabled ACG to function as a start codon for Rep78/68 translation. The data further indicate that the function of the AAV helper construct (pAAV-RC), that is in current widespread use for rAAV production, may be improved by replacement of its AAV2 unrelated heterologous sequence with the native AAV2 p5 promoter. CONCLUSION: Taken together, the data demonstrate an interplay between the start codon and upstream regulatory sequences in the regulation of Rep78/68 and indicate that selective mutations in Rep78/68 regulatory elements may serve to augment the therapeutic value of rAAV vectors. PMID- 19128487 TI - Disabling osteomalacia and myopathy as the only presenting features of celiac disease: a case report. AB - BACKGROUND: Celiac disease is an autoimmune inflammatory disease of the small intestine precipitated by the ingestion of gluten, a component of wheat protein, in genetically susceptible persons. Classically, the disease manifests with diarrhea, weight loss and anemia. There are very few reports of osteomalacia as the presenting symptom, and even fewer of osteomalacia as the only symptom of celiac disease at presentation. CASE PRESENTATION: In this case report we describe a 40 year-old patient who presented with 6 months history of progressively worsening and disabling osteomalacia as the only presentation of a celiac disease. With adherence to a gluten-free diet and calcium-vitamin D supplementation, the patient's condition improved remarkably within three months and she was able to walk pain free. CONCLUSION: Celiac disease frequently is under diagnosed or misdiagnosed which results in unnecessary morbidity such as disabling osteomalacia. Therefore, early diagnosis of celiac disease is important because the symptoms completely resolve with adequate treatment. PMID- 19128488 TI - Strangulated inguinal hernia due to an omental band adhesion within the hernial sac: a case report. AB - INTRODUCTION: Strangulated Inguinal hernia is one of the most common surgical emergencies dealt with by surgeons worldwide. Usually the narrow internal inguinal ring or the external inguinal ring is the site of constriction of the viscus, which forms the content of the hernia resulting in strangulation. We report a rare case of strangulated inguinal hernia where the constricting element is not the internal or external inguinal ring, but an omental band adhesion causing closed loop small bowel obstruction and gangrene within the hernial sac in the inguinal canal. CASE REPORT: A 56-year-old Caucasian gentleman presented to us with a 6 hours history of non-reducible tender lump in his right groin. His groin was explored urgently under general anaesthesia and was found to have an omental band adhesion causing closed loop small bowel obstruction with gangrene within the hernial sac in the inguinal canal with a wide internal inguinal ring. Gangrenous small bowel was resected and primary anastomosis was performed through the same inguinal incision. CONCLUSION: Strangulation of the inguinal hernial content is usually due to the tight constriction at the level of internal inguinal ring or at external inguinal ring. Uncommonly strangulation of the contents can occur due to other causes like omental band adhesion. Anyone presenting with clinical features of strangulated inguinal hernia with small bowel obstruction mandates prompt exploration of the inguinal canal. Although it may not change the treatment approach, one should be aware about this special entity. Resection of the gangrenous small bowel and primary anastomosis can be safely performed through the same inguinal incision. PMID- 19128489 TI - Regional disparities in infant mortality in Canada: a reversal of egalitarian trends. AB - BACKGROUND: Although national health insurance plans and social programs introduced in the 1960s led to reductions in regional disparities in infant mortality in Canada, it is unclear if such patterns prevailed in the 1990s when the health care and related systems were under fiscal duress. This study examined regional patterns of change in infant mortality in Canada in recent decades. METHODS: We analysed regional changes in crude infant mortality rates and in infant mortality rates among live births with a birth weight >or= 500 g and >or= 1,000 g in Canada from 1945 to 2002. Associations between baseline infant mortality rates in the provinces and territories (e.g., in 1985-89) and the change observed in infant mortality rates over the subsequent period (e.g., between 1985-89 and 1995-99) were assessed using Spearman's rank correlation coefficient. Trends in regional disparities were also assessed by calculating period-specific rate ratios between provinces/territories with the highest versus the lowest infant mortality. RESULTS: Provincial/territorial infant mortality rates in 1945-49 were not correlated with changes in infant mortality over the next 10 years (rho = 0.01, P = 0.99). However, there was a strong negative correlation between infant mortality rates in 1965-69 and the subsequent decline in infant mortality (rho = - 0.85, P = 0.002). Provinces/territories with higher infant mortality rates in 1965-69 (Northwest Territories 64.7 vs British Columbia 20.7 per 1,000 live births) experienced relatively larger reductions in infant mortality between 1965-69 and 1975-79 (53.7% decline in the Northwest Territories vs a 36.6% decline in British Columbia). This pattern was reversed in the more recent decades. Provinces/territories with higher infant mortality rates >or= 500 g in 1985-89 experience relatively smaller reductions in infant mortality between 1985-89 and 2000-02 (rho = 0.82, P = 0.004). The infant mortality >or= 500 g rate ratio (contrasting the province/territory with the highest versus lowest infant mortality) was 3.2 in 1965-69, 2.4 in 1975-79, 2.2 in 1985-89, 3.1 in 1995-99 and 4.1 in 2000-02. CONCLUSION: Fiscal constraints in the 1990s led to a reversal of provincial/territorial patterns of change in infant mortality in Canada and to an increase in regional health disparities. PMID- 19128490 TI - Tardy posterior interosseous nerve palsy associated with radial head fracture: a case report. AB - BACKGROUND: A 55 year old gentleman presented with tardy posterior interosseous nerve palsy associated with radial head fracture. CASE PRESENTATION: The patient developed symptoms of wrist drop 24 hours after the injury whilst awaiting surgery for his Mason III radial head fracture. EMG studies confirmed the presence of a posterior interosseous nerve lesion. Open exploration revealed oedematous soft tissues surrounding the nerve. The proximity of the nerve to the fracture and its course through the arcade of Frohse make it susceptible to injury, from the initial traumatic event and the following oedema. CONCLUSION: A full recovery occurred after exploration and decompression of the nerve. PMID- 19128491 TI - Giant lymphatic cyst of omentum: a case report. AB - BACKGROUND: Omental cysts are rare abdominal lesions and are difficult to diagnose. Mostly they are detected incidentally during imaging studies performed for unrelated reasons. CASE PRESENTATION: Presentation can be both acute and chronic. Acute presentations are usually due to complication in cyst. Imaging is helpful in excluding other causes of lump abdomen. We encountered a case of giant lymphatic cyst presenting with abdominal swelling, clinically mimicking huge ovarian cyst. CONCLUSION: The goal of surgical therapy is complete excision of the cyst, and Omental cysts can be removed without endangering the adjacent bowel. PMID- 19128493 TI - Telangiectatic adenoma - computed tomography and magnetic resonance findings: a case report and review of the literature. AB - Telangiectatic adenoma is a new classification of a hepatic lesion. It was previously named telangiectatic focal nodular hyperplasia but it is in fact true adenoma with telangiectatic features. We report here a case of telangiectatic adenoma in a 72-year-old woman. The image features are lack of a central scar, a heterogeneous lesion, hyperintensity in T1-weighted MR images, strong hyperintensity in T2-weighted MR images, and persistent contrast enhancement in delayed-phase contrast-enhanced CT or T1-weighted MR images. It is a monoclonal lesion with potential of malignancy. The treatment of telangiectatic adenoma is surgery, the same way as hepatic adenoma. Focal nodular hyperplasia may be managed by clinical follow-up alone. PMID- 19128492 TI - Asymmetrical distribution of non-conserved regulatory sequences at PHOX2B is reflected at the ENCODE loci and illuminates a possible genome-wide trend. AB - BACKGROUND: Transcriptional regulatory elements are central to development and interspecific phenotypic variation. Current regulatory element prediction tools rely heavily upon conservation for prediction of putative elements. Recent in vitro observations from the ENCODE project combined with in vivo analyses at the zebrafish phox2b locus suggests that a significant fraction of regulatory elements may fall below commonly applied metrics of conservation. We propose to explore these observations in vivo at the human PHOX2B locus, and also evaluate the potential evidence for genome-wide applicability of these observations through a novel analysis of extant data. RESULTS: Transposon-based transgenic analysis utilizing a tiling path proximal to human PHOX2B in zebrafish recapitulates the observations at the zebrafish phox2b locus of both conserved and non-conserved regulatory elements. Analysis of human sequences conserved with previously identified zebrafish phox2b regulatory elements demonstrates that the orthologous sequences exhibit overlapping regulatory control. Additionally, analysis of non-conserved sequences scattered over 135 kb 5' to PHOX2B, provides evidence of non-conserved regulatory elements positively biased with close proximity to the gene. Furthermore, we provide a novel analysis of data from the ENCODE project, finding a non-uniform distribution of regulatory elements consistent with our in vivo observations at PHOX2B. These observations remain largely unchanged when one accounts for the sequence repeat content of the assayed intervals, when the intervals are sub-classified by biological role (developmental versus non-developmental), or by gene density (gene desert versus non-gene desert). CONCLUSION: While regulatory elements frequently display evidence of evolutionary conservation, a fraction appears to be undetected by current metrics of conservation. In vivo observations at the PHOX2B locus, supported by our analyses of in vitro data from the ENCODE project, suggest that the risk of excluding non-conserved sequences in a search for regulatory elements may decrease as distance from the gene increases. Our data combined with the ENCODE data suggests that this may represent a genome wide trend. PMID- 19128494 TI - Inpatient charges among HIV/AIDS patients in Rhode Island from 2000-2004. AB - BACKGROUND: Inpatient HIV/AIDS charges decreased from 1996-2000. This decrease was mainly attributable to treatment of HIV/AIDS patients with Highly Active Antiretroviral Therapy (HAART). This study aims to evaluate the trend in inpatient charges from 2000-2004. METHODS: Rhode Island Hospital Discharge Data (HDD) from 2000 to 2004 was used. International Classification of Disease (ICD-9) diagnosis code 042-044 was used to identify HIV/AIDS admissions. The final study population included 1927 HIV/AIDS discharges. We used a multivariable linear regression model to examine the factors associated with inflation adjusted inpatient charges. RESULTS: We found a significant increase in inpatient charges from 2000-2004 after adjusting for length of stay (LOS), gender, age, race and point of entry for hospitalization. In addition to calendar year, LOS, gender and race were also associated with inpatient charges. CONCLUSION: HIV/AIDS inpatient charges increased after adjusting for inflation despite earlier studies that showed a decline. Our results have implications for uninsured, as well as insured HIV/AIDS patients who do not have a medical plan that covers their charges sufficiently. Future research should investigate what factors are contributing to rising inpatient charges among HIV/AIDS patients. PMID- 19128495 TI - Flux balance analysis of primary metabolism in Chlamydomonas reinhardtii. AB - BACKGROUND: Photosynthetic organisms convert atmospheric carbon dioxide into numerous metabolites along the pathways to make new biomass. Aquatic photosynthetic organisms, which fix almost half of global inorganic carbon, have great potential: as a carbon dioxide fixation method, for the economical production of chemicals, or as a source for lipids and starch which can then be converted to biofuels. To harness this potential through metabolic engineering and to maximize production, a more thorough understanding of photosynthetic metabolism must first be achieved. A model algal species, C. reinhardtii, was chosen and the metabolic network reconstructed. Intracellular fluxes were then calculated using flux balance analysis (FBA). RESULTS: The metabolic network of primary metabolism for a green alga, C. reinhardtii, was reconstructed using genomic and biochemical information. The reconstructed network accounts for the intracellular localization of enzymes to three compartments and includes 484 metabolic reactions and 458 intracellular metabolites. Based on BLAST searches, one newly annotated enzyme (fructose-1,6-bisphosphatase) was added to the Chlamydomonas reinhardtii database. FBA was used to predict metabolic fluxes under three growth conditions, autotrophic, heterotrophic and mixotrophic growth. Biomass yields ranged from 28.9 g per mole C for autotrophic growth to 15 g per mole C for heterotrophic growth. CONCLUSION: The flux balance analysis model of central and intermediary metabolism in C. reinhardtii is the first such model for algae and the first model to include three metabolically active compartments. In addition to providing estimates of intracellular fluxes, metabolic reconstruction and modelling efforts also provide a comprehensive method for annotation of genome databases. As a result of our reconstruction, one new enzyme was annotated in the database and several others were found to be missing; implying new pathways or non-conserved enzymes. The use of FBA to estimate intracellular fluxes also provides flux values that can be used as a starting point for rational engineering of C. reinhardtii. From these initial estimates, it is clear that aerobic heterotrophic growth on acetate has a low yield on carbon, while mixotrophically and autotrophically grown cells are significantly more carbon efficient. PMID- 19128497 TI - Chromatid recommensuration after segmental duplication. AB - BACKGROUND: Midsegment duplication (dup) of chromatid arms may be symmetric or asymmetric. It can be argued that every dup should yield a discommensured RC with (a) loss of at least one duplicated unit to the template counterpart and; (b) deletion of all sections of the replicating chromatid arm that are distal to both the gap left by the duplicating process and the segment closest to the centromere. HYPOTHESIS: Mechanisms capable of recommensuring the stack of chromatids after topological shifts of duplicated units (dups) are discussed. The mechanics might fail in few cases, which are discussed in terms of statistics and scalability. CONCLUSION: The dynamics of the highly non-linear processes discussed here may be relevant to duplications of smaller (epsilon) subunits such as telomeric units within malignant genomes. PMID- 19128496 TI - Complex organizational structure of the genome revealed by genome-wide analysis of single and alternative promoters in Drosophila melanogaster. AB - BACKGROUND: The promoter is a critical necessary transcriptional cis-regulatory element. In addition to its role as an assembly site for the basal transcriptional apparatus, the promoter plays a key part in mediating temporal and spatial aspects of gene expression through differential binding of transcription factors and selective interaction with distal enhancers. Although many genes have multiple promoters, little attention has been focused on how these relate to one another; nor has much study been directed at relationships between promoters of adjacent genes. RESULTS: We have undertaken a systematic investigation of Drosophila promoters. We divided promoters into three groups: unique promoters, first alternative promoters (the most 5' of a gene's multiple promoters), and downstream alternative promoters (the remaining alternative promoters 3' to the first). We observed distinct nucleotide distribution and sequence motif preferences among these three classes. We also investigated the promoters of neighboring genes and found that a greater than expected number of adjacent genes have similar sequence motif profiles, which may allow the genes to be regulated in a coordinated fashion. Consistent with this, there is a positive correlation between similar promoter motifs and related gene expression profiles for these genes. CONCLUSIONS: Our results suggest that different regulatory mechanisms may apply to each of the three promoter classes, and provide a mechanism for "gene expression neighborhoods," local clusters of co-expressed genes. As a whole, our data reveal an unexpected complexity of genomic organization at the promoter level with respect to both alternative and neighboring promoters. PMID- 19128498 TI - Pastoralism and delay in diagnosis of TB in Ethiopia. AB - BACKGROUND: Tuberculosis (TB) is a major public health problem in the Horn of Africa with Ethiopia being the most affected where TB cases increase at the rate of 2.6% each year. One of the main contributing factors for this rise is increasing transmission due to large number of untreated patients, serving as reservoirs of the infection within the communities. Reduction of the time between onset of TB symptoms to diagnosis is therefore a prerequisite to bring the TB epidemic under control. The aim of this study was to measure duration of delay among pastoralist TB patients at TB management units in Somali Regional State (SRS) of Ethiopia. METHODS: A cross sectional study of 226 TB patients with pastoralist identity was conducted in SRS of Ethiopia from June to September 2007. Patients were interviewed using questionnaire based interview. Time between onset of TB symptoms and first visit to a professional health care provider (patient delay), and the time between first visits to the professional health care provider to the date of diagnosis (medical provider's delay) were analyzed. Both pulmonary and extrapulmonary TB patients were included in the study. RESULT: A total of 226 pastoralist TB patients were included in this study; 93 (41.2%) were nomadic pastoralists and 133 (58.8%) were agro-pastoralists. Median patient delay was found to be 60 days with range of 10-1800 days (83 days for nomadic pastoralists and 57 days for agro-pastoralists). Median health care provider's delay was 6 days and median total delay was 70 days in this study. Patient delay constituted 86% of the total delay. In multivariate logistic regression analysis, nomadic pastoralism (aOR. 2.69, CI 1.47-4.91) and having low biomedical knowledge on TB (aOR. 2.02, CI 1.02-3.98) were significantly associated with prolonged patient delay. However, the only observed risk factor for very long patient delay >120 days was distance to health facility (aOR.4.23, CI 1.32-13.54). Extra pulmonary TB was the only observed predictor for health care providers' delay (aOR. 3.39, CI 1.68-6.83). CONCLUSION: Patient delay observed among pastoralist TB patients in SRS is one of the highest reported so far from developing countries, exceeding two years in some patients. This long patient delay appears to be associated with patient's inadequate knowledge of the disease and distance to health care facility with nomadic pastoralists being the most affected. Regional TB control programmes need to consider the exceptional circumstances of pastoralists, to maximise their access to TB services. PMID- 19128499 TI - Functional morphology and integration of corvid skulls - a 3D geometric morphometric approach. AB - BACKGROUND: Sympatric corvid species have evolved differences in nesting, habitat choice, diet and foraging. Differences in the frequency with which corvid species use their repertoire of feeding techniques is expected to covary with bill-shape and with the frontal binocular field. Species that frequently probe are expected to have a relatively longer bill and more sidewise oriented orbits in contrast to species that frequently peck. We tested this prediction by analyzing computed tomography scans of skulls of six corvid species by means of three-dimensional geometric morphometrics. We (1) explored patterns of major variation using principal component analysis, (2) compared within and between species relationships of size and shape and (3) quantitatively compared patterns of morphological integration between bill and cranium by means of partial least squares (singular warp) analysis. RESULTS: Major shape variation occurs at the bill, in the orientation of orbits, in the position of the foramen magnum and in the angle between bill and cranium. The first principal component correlated positively with centroid-size, but within-species allometric relationships differed markedly. Major covariation between the bill and cranium lies in the difference in orbit orientation relative to bill-length and in the angle between bill and cranium. CONCLUSION: Corvid species show pronounced differences in skull shape, which covary with foraging mode. Increasing bill-length, bill-curvature and sidewise orientation of the eyes is associated with an increase in the observed frequency in probing (vice versa in pecking). Hence, the frequency of probing, bill-length, bill-curvature and sidewise orientation of the eyes is progressively increased from jackdaw, to Eurasian jay, to black-billed magpie, to hooded crow, to rook and to common raven (when feeding on carcasses is considered as probing). Our results on the morphological integration suggest that most of the covariation between bill and cranium is due to differences in the topography of the binocular fields and the projection of the bill-tip therein, indicating the importance of visual fields to the foraging ecology of corvids. PMID- 19128500 TI - A dosimetric comparison of different treatment plans of palliative spinal bone irradiation: analysis of dose coverage with respect to ICRU 50 Report. AB - BACKGROUND: This study aimed to analyze three-dimensional (3D) dosimetric data of conventional two-dimensional (2D) palliative spinal bone irradiation using different reference points and treatment plans with respect to the International Commission on Radiation Units and Measurements (ICRU) Report 50. METHODS: Forty five simulation CT scans of 39 patients previously treated for thoraco-lumbar spinal bone metastases were used. Three different treatment plans were created: (1) single posterior field plans using the ICRU reference points (ICRUrps); (2) single posterior field plans using the International Bone Metastasis Consensus Working Party reference points (IBMCrps); (3) two opposed anterior-posterior (AP PA) field plans using the ICRUrps. The intended dose range for planning target volume (PTV) was 90% to 110% of the prescribed dose for AP-PA field plans. Cumulative dose-volume histograms were generated for each plan, and minimum, maximum and mean doses to the PTV, medulla spinalis, esophagus and intestines were analyzed. RESULTS: The mean percentages of minimum, maximum and mean PTV doses +/- standard deviation were, respectively, 91 +/- 1.3%, 108.8 +/- 1.3% and 99.7 +/- 1.3% in AP-PA field plans; 77.3 +/- 2.6%, 122.2 +/- 4.3% and 99.8 +/- 2.6% in ICRUrp single field plans; and 83.7 +/- 3.3%, 133.9 +/- 7.1% and 108.8 +/ 3.3% in IBMCrp single field plans. Minimum doses of both single field plans were significantly lower (p < 0.001) while maximum doses were significantly higher (p < 0.001) than AP-PA field plans. Minimum, maximum and mean doses were higher in IBMCrp single field plans than in ICRUrp single field plans (p < 0.001). The mean medulla spinalis doses were lower in AP-PA field plans than single posterior field plans (p < 0.001). Maximum doses for medulla spinalis were higher than 120% of the prescribed dose in 22 of 45 (49%) IBMCrp single field plans. Mean esophagus and intestinal doses were higher (p < 0.001) in AP-PA field plans than single field plans, however, less than 95% of the prescribed dose. CONCLUSION: In palliative spinal bone irradiation, 2D conventional single posterior field radiotherapy did not accomplish the ICRU Report 50 recommendations for PTV dose distribution, while the AP-PA field plans did achieve the intended dose ranges with a homogenous distribution and reasonable doses to the medulla spinalis, esophagus and intestines. PMID- 19128502 TI - Mosaic analysis of stem cell function and wound healing in the mouse corneal epithelium. AB - BACKGROUND: The mouse corneal epithelium is a continuously renewing 5-6 cell thick protective layer covering the corneal surface, which regenerates rapidly when injured. It is maintained by peripherally located limbal stem cells (LSCs) that produce transient amplifying cells (TACs) which proliferate, migrate centripetally, differentiate and are eventually shed from the epithelial surface. LSC activity is required both for normal tissue maintenance and wound healing. Mosaic analysis can provide insights into LSC function, cell movement and cell mixing during tissue maintenance and repair. The present study investigates cell streaming during corneal maintenance and repair and changes in LSC function with age. RESULTS: The initial pattern of corneal epithelial patches in XLacZ+/- X inactivation mosaics was replaced after birth by radial stripes, indicating activation of LSCs. Stripe patterns (clockwise, anticlockwise or midline) were independent between paired eyes. Wound healing in organ culture was analysed by mosaic analysis of XLacZ+/- eyes or time-lapse imaging of GFP mosaics. Both central and peripheral wounds healed clonally, with cells moving in from all around the wound circumference without significant cell mixing, to reconstitute striping patterns. Mosaic analysis revealed that wounds can heal asymmetrically. Healing of peripheral wounds produced stripe patterns that mimicked some aberrant striping patterns observed in unwounded corneas. Quantitative analysis provided no evidence for an uneven distribution of LSC clones but showed that corrected corneal epithelial stripe numbers declined with age (implying declining LSC function) but stabilised after 39 weeks. CONCLUSION: Striping patterns, produced by centripetal movement, are defined independently and stochastically in individual eyes. Little cell mixing occurs during the initial phase of wound healing and the direction of cell movement is determined by the position of the wound and not by population pressure from the limbus. LSC function declines with age and this may reflect reduced LSCs numbers, more quiescent LSCs or a reduced ability of older stem cells to maintain tissue homeostasis. The later plateau of LSC function might indicate the minimum LSC function that is sufficient for corneal epithelial maintenance. Quantitative and temporal mosaic analyses provide new possibilities for studying stem cell function, tissue maintenance and repair. PMID- 19128501 TI - Phase II trial of Modified Vaccinia Ankara (MVA) virus expressing 5T4 and high dose Interleukin-2 (IL-2) in patients with metastatic renal cell carcinoma. AB - BACKGROUND: Interleukin-2 (IL-2) induces durable objective responses in a small cohort of patients with metastatic renal cell carcinoma (RCC) but the antigen(s) responsible for tumor rejection are not known. 5T4 is a non-secreted membrane glycoprotein expressed on clear cell and papillary RCCs. A modified vaccinia virus Ankara (MVA) encoding 5T4 was tested in combination with high-dose IL-2 to determine the safety, objective response rate and effect on humoral and cell mediated immunity. METHODS: 25 patients with metastatic RCC who qualified for IL 2 were eligible and received three immunizations every three weeks followed by IL 2 (600,000 IU/kg) after the second and third vaccinations. Blood was collected for analysis of humoral, effector and regulatory T cell responses. RESULTS: There were no serious vaccine-related adverse events. While no objective responses were observed, three patients (12%) were rendered disease-free after nephrectomy or resection of residual metastatic disease. Twelve patients (48%) had stable disease which was associated with improved median overall survival compared to patients with progressive disease (not reached vs. 28 months, p = 0.0261). All patients developed 5T4-specific antibody responses and 13 patients had an increase in 5T4-specific T cell responses. Although the baseline frequency of Tregs was elevated in all patients, those with stable disease showed a trend toward increased effector CD8+ T cells and a decrease in Tregs. CONCLUSION: Vaccination with MVA-5T4 did not improve objective response rates of IL-2 therapy but did result in stable disease associated with an increase in the ratio of 5T4 specific effector to regulatory T cells in selected patients. TRIAL REGISTRATION NUMBER: ISRCTN83977250. PMID- 19128503 TI - Effect of sirolimus on urinary bladder cancer T24 cell line. AB - BACKGROUND: Sirolimus is recently reported to have antitumour effects on a large variety of cancers. The present study was performed to investigate sirolimus's ability to inhibit growth in T24 bladder cancer cells. METHODS: T24 bladder cancer cells were treated with various concentrations of sirolimus. MTT assay was used to evaluate the proliferation inhibitory effect on T24 cell line. The viability of T24 cell line was determined by Trypan blue exclusion analysis. RESULTS: Sirolimus inhibits the growth of bladder carcinoma cells and decreases their viability. Significant correlations were found between cell proliferation and sirolimus concentration (r = 0.830; p < 0.01) as well as between cell viability and sirolimus concentration (r = -0.896; p < 0.01). CONCLUSION: Sirolimus has an anti-proliferation effect on the T24 bladder carcinoma cell line. The information from our results is useful for a better understanding sirolimus's anti-proliferative activity in the T24 bladder cancer cell line. PMID- 19128504 TI - A multiple-method approach reveals a declining amount of chloroplast DNA during development in Arabidopsis. AB - BACKGROUND: A decline in chloroplast DNA (cpDNA) during leaf maturity has been reported previously for eight plant species, including Arabidopsis thaliana. Recent studies, however, concluded that the amount of cpDNA during leaf development in Arabidopsis remained constant. RESULTS: To evaluate alternative hypotheses for these two contradictory observations, we examined cpDNA in Arabidopsis shoot tissues at different times during development using several methods: staining leaf sections as well as individual isolated chloroplasts with 4',6-diamidino-2-phenylindole (DAPI), real-time quantitative PCR with DNA prepared from total tissue as well as from isolated chloroplasts, fluorescence microscopy of ethidium-stained DNA molecules prepared in gel from isolated plastids, and blot-hybridization of restriction-digested total tissue DNA. We observed a developmental decline of about two- to three-fold in mean DNA per chloroplast and two- to five-fold in the fraction of cellular DNA represented by chloroplast DNA. CONCLUSION: Since the two- to five-fold reduction in cpDNA content could not be attributed to an artifact of chloroplast isolation, we conclude that DNA within Arabidopsis chloroplasts is degraded in vivo as leaves mature. PMID- 19128505 TI - Large-scale prediction of long disordered regions in proteins using random forests. AB - BACKGROUND: Many proteins contain disordered regions that lack fixed three dimensional (3D) structure under physiological conditions but have important biological functions. Prediction of disordered regions in protein sequences is important for understanding protein function and in high-throughput determination of protein structures. Machine learning techniques, including neural networks and support vector machines have been widely used in such predictions. Predictors designed for long disordered regions are usually less successful in predicting short disordered regions. Combining prediction of short and long disordered regions will dramatically increase the complexity of the prediction algorithm and make the predictor unsuitable for large-scale applications. Efficient batch prediction of long disordered regions alone is of greater interest in large-scale proteome studies. RESULTS: A new algorithm, IUPforest-L, for predicting long disordered regions using the random forest learning model is proposed in this paper. IUPforest-L is based on the Moreau-Broto auto-correlation function of amino acid indices (AAIs) and other physicochemical features of the primary sequences. In 10-fold cross validation tests, IUPforest-L can achieve an area of 89.5% under the receiver operating characteristic (ROC) curve. Compared with existing disorder predictors, IUPforest-L has high prediction accuracy and is efficient for predicting long disordered regions in large-scale proteomes. CONCLUSION: The random forest model based on the auto-correlation functions of the AAIs within a protein fragment and other physicochemical features could effectively detect long disordered regions in proteins. A new predictor, IUPforest-L, was developed to batch predict long disordered regions in proteins, and the server can be accessed from http://dmg.cs.rmit.edu.au/IUPforest/IUPforest L.php. PMID- 19128506 TI - Distribution of antibody titer against Salmonella enterica among healthy individuals in nepal. AB - BACKGROUND: Enteric fever is an endemic problem in Nepal and Widal agglutination test is widely used for its diagnosis but a normal baseline titer in healthy population and cutoff values have not been established. METHODS: We measured average baseline antibody titers against "O" and "H" antigens of Salmonella enterica serotype Typhi and "H" antigens of serotypes Paratyphi A and Paratyphi B among apparently healthy blood donors in Nepal. The antibody titers were measured using Standard Widal Confirmatory Quantitative Tube test. RESULTS: Among the 100 blood samples collected from healthy volunteers, 62 individuals had significant antibody titers (> or = 1:20) against one of the four antigens against S. enterica. Among 54 samples with an anti-O titer against serotype Typhi, 15 and 36 samples had titers of > or = 1:60 and > or = 1:40, respectively. A significant proportion (12% of all) had anti-O titer of > or = 1:80. Similarly, among the 59 samples demonstrating anti-H titers of > or = 1:20 to S. enterica serotype Typhi, 29 had a titer of > or = 1:80 and 12 had 1:160. For S. enterica serotypes Paratyphi A and B, anti-H titers of > or = 1:20 were found only in 12% and 3%, respectively, of all samples tested. CONCLUSION: When a single Widal agglutination titer is used for the diagnosis of enteric fever, it will be more appropriate to change the currently used cutoff levels against S. enterica serotype Typhi to > 1:80 for anti-O and > 1:160 for anti-H titers for Nepal. PMID- 19128507 TI - Severe cutaneous toxicity following treatment with radiotherapy and cetuximab: a case report. AB - While the addition of cetuximab to radiotherapy improves clinical outcomes in locoregionally advanced head and neck squamous cell cancers, there are a small number of reports of severe radiation dermatitis occurring with this therapeutic combination. We present the case of a 69 year old male who developed severe radiation dermatitis following treatment with cetuximab and radiotherapy for a locoregionally advanced head and neck squamous cell cancer. PMID- 19128508 TI - TCP: a tool for designing chimera proteins based on the tertiary structure information. AB - BACKGROUND: Chimera proteins are widely used for the analysis of the protein protein interaction region. One of the major issues is the epitope analysis of the monoclonal antibody. In the analysis, a continuous portion of an antigen is sequentially substituted into a different sequence. This method works well for an antibody recognizing a linear epitope, but not for that recognizing a discontinuous epitope. Although the designing the chimera proteins based on the tertiary structure information is required in such situations, there is no appropriate tool so far. RESULTS: In light of the problem, we developed a tool named TCP (standing for a Tool for designing Chimera Proteins), which extracts some sets of mutually orthogonal cutting surfaces for designing chimera proteins using a genetic algorithm. TCP can also incorporate and consider the solvent accessible surface area information calculated by a DSSP program. The test results of our method indicate that the TCP is robust and applicable to various shapes of proteins. CONCLUSION: We developed TCP, a tool for designing chimera proteins based on the tertiary structure information. TCP is robust and possesses several favourable features, and we believe it is a useful tool for designing chimera proteins. TCP is freely available as an additional file of this manuscript for academic and non-profit organization. PMID- 19128509 TI - Mutation analysis of the Gadd45 gene at exon 4 in atypical fibroxanthoma. AB - BACKGROUND: Atypical fibroxanthoma (AFX) histologically mimics high-grade sarcoma in the skin, although it follows a benign clinical course. AFX occurs in the sun exposed skin and for this reason, an association with ultraviolet light has long been suspected. Bax and Gadd45 are p53 effector proteins. Bax is a programmed cell death protein and belongs to the Bcl-2 family. Gadd45 is a multifunctional DNA damage-inducible gene associated with the process of DNA damage. METHODS: Immunohistochemical expression of Bax was analyzed in 7 cases of AFX, and in 7 cases of benign fibrous histiocytoma (BFH) used as a comparison. The expression pattern of Bax was compared to previously reported p53 and Gadd45 expressions in a correspondent series. Mutation of the Gadd45 gene at exon 4 was also analyzed in AFX. RESULTS: AFX and BFH showed immunoreactivities respectively for Bax (3/7, 0/7), Gadd45 (4/7, 1/7) and p53 (2/7, 0/7). There was no exact correlation between p53 expression and Bax or Gadd45 expression. However, the pattern of expression between Bax and Gadd45 was also the same, with the exception of one case. No mutation of the Gadd45 gene at exon 4 was observed in a series of 6 AFX cases where DNA was available (0/6). CONCLUSION: These results suggest a possible association between Bax and Gadd45 in AFX, and may refute any possibility of dysfunction of Gadd45 in terms of gene mutation, at least at exon 4 of the Gadd45 gene. PMID- 19128510 TI - MDM2 is a novel E3 ligase for HIV-1 Vif. AB - The human immunodeficiency virus type 1 (HIV-1) Vif plays a crucial role in the viral life cycle by antagonizing a host restriction factor APOBEC3G (A3G). Vif interacts with A3G and induces its polyubiquitination and subsequent degradation via the formation of active ubiquitin ligase (E3) complex with Cullin5 ElonginB/C. Although Vif itself is also ubiquitinated and degraded rapidly in infected cells, precise roles and mechanisms of Vif ubiquitination are largely unknown. Here we report that MDM2, known as an E3 ligase for p53, is a novel E3 ligase for Vif and induces polyubiquitination and degradation of Vif. We also show the mechanisms by which MDM2 only targets Vif, but not A3G that binds to Vif. MDM2 reduces cellular Vif levels and reversely increases A3G levels, because the interaction between MDM2 and Vif precludes A3G from binding to Vif. Furthermore, we demonstrate that MDM2 negatively regulates HIV-1 replication in non-permissive target cells through Vif degradation. These data suggest that MDM2 is a regulator of HIV-1 replication and might be a novel therapeutic target for anti-HIV-1 drug. PMID- 19128511 TI - Inhibition of PI3K increases oxaliplatin sensitivity in cholangiocarcinoma cells. AB - BACKGROUND: Resistance of cholangiocarcinoma to chemotherapy is a major problem in cancer treatment. The mechanism of resistance is believed to involve phosphoinositide-3- kinase (PI3K)/Akt activation. Although the platinum containing compound oxaliplatin has been extensively used in the treatment of several solid tumors, recent data regarding its use to treat cholangiocarcinoma are ambiguous. Oxaliplatin resistance in this disease could potentially involve PI3K pathways. We, therefore, examined the effects of PI3K pathways in cholangiocarcinoma cells in modulating oxaliplatin resistance. RESULTS: After exposing the cholangiocarcinoma cell lines RMCCA1 and KKU100 to oxaliplatin, the levels of Akt and mTOR phosphorylation increased, as shown by western blot analysis. The WST-1 cell proliferation assay showed increased inhibition of cell growth under high concentrations of oxaliplatin. The combination of oxaliplatin with LY294002, an inhibitor of PI3K, resulted in a remarkable arrest of cell proliferation. Deactivation of mTOR by RAD001 was also synergistic with oxaliplatin, although to a lesser extent. The combination of oxaliplatin and a PI3K inhibitor also resulted in a significant induction of apoptosis, as demonstrated by the TUNEL assay. CONCLUSION: Activation of PI3K might protect cholangiocarcinoma cells from oxaliplatininduced cytotoxicity. Although the inhibition of PI3K and the inhibition of mTOR both enhance oxaliplatin-induced cytotoxicity, PI3K inhibition has a greater effect. Targeting the PI3K pathway may be a useful approach to improve the chemotherapeutic sensitivity of cholangiocarcinoma. PMID- 19128512 TI - The differential expressions of 78-kDa glucose-regulated protein of infiltrating plasma cells in peripheral joints with the histopathological variants of rheumatoid synovitis. AB - INTRODUCTION: The local production of pathogenic autoantibodies by plasma cells in synovium is one of the hallmarks of rheumatoid arthritis (RA). There may be a potential link between ectopic lymphoid neogenesis and the local autoimmunity in rheumatoid synovium. The unfolded protein response (UPR) has key roles in the development and maintenance of plasma cells secreting immunoglobulin. This study was designed to explore the potential links between the activation of the UPR of infiltrating plasma cells in inflamed peripheral joints and the histopathological variants of rheumatoid synovitis as well as the local production of pathogenic autoantibodies. METHODS: The variants of rheumatoid synovium were histopathologically classified into follicular and diffuse synovitis. Immunohistochemical and double-immunofluorescent stainings were performed to detect the expression of 78-kDa glucose-regulated protein (GRP78), a marker of activation of the UPR, in infiltrating plasma cells of synovium, and flow cytometry and immunoblotting analyses were performed to quantify GRP78 in plasma cells of synovial fluid in inflamed peripheral joints of RA. The detections were also taken in osteoarthritis (OA) as controls. The synovial fluid levels of anti cyclic citrullinated peptide antibodies (anti-CCP) (IgG) were quantified with the enzyme-linked immunosorbent assay and corrected to those of total IgG in RA. RESULTS: Expressions of GRP78 were more intensive in infiltrating plasma cells in RA synovium relative to those in OA synovium (P < 0.001) and in synovium with follicular synovitis relative to that with diffuse synovitis (P < 0.001). Analyses by flow cytometry and immunoblotting showed that there was a significant upregulation of GRP78 of plasma cells from synovial fluid of RA compared with that of OA (P < 0.05) and from synovial fluid of follicular synovitis relative to that of diffuse synovitis (P < 0.05). Moreover, a positive relationship between the expression of GRP78 of plasma cells from synovial fluid and the corrected synovial levels of anti-CCP (IgG) was seen in RA (P < 0.001). CONCLUSIONS: There may be a link between enhanced activation of the UPR of plasma cells and ectopic lymphoid neogenesis as well as the local production of anti-CCP (IgG) in inflamed peripheral joints of RA. PMID- 19128514 TI - Fibrous dysplasia - differential diagnosis of cystic lesions in the proximal femur:a case report. AB - BACKGROUND: We are reporting about the case of a 22-year old woman, who made a first visit as an outpatient with pain that arose in acute form in the area of the left groin. The patient history did not indicate a preceding trauma, or an inflammatory or malignant disease. CASE PRESENTATION: Conventional X-ray showed the image of a cystic lesion in the area of the neck of the thigh [collum femoris] with pathological fracture in the area of the calcar. The MRT produced the diagnosis of an aneurismal bone cyst, in which the presence of juvenile bone cyst or a giant cell tumor could be taken into consideration by differential diagnosis.Owing to the pathological fracture, repair by operation of the lesion was carried out by means of spongioplastic surgery and internal fixation (DHS). CONCLUSION: Histology produced characteristic findings fitting the condition of fibrous dysplasia, which was not included among the primary causes in the considerations offered by differential diagnosis.Therefore, this case is a good example of the fact that fibrous dysplasia must be taken into account when using differential diagnosis as one of the cystic lesions. PMID- 19128513 TI - Glycoproteomic analysis of two mouse mammary cell lines during transforming growth factor (TGF)-beta induced epithelial to mesenchymal transition. AB - BACKGROUND: TGF-beta acts as an antiproliferative factor in normal epithelial cells and at early stages of oncogenesis. However, later in tumor development TGF beta can become tumor promoting through mechanisms including the induction of epithelial-to-mesenchymal transition (EMT), a process that is thought to contribute to tumor progression, invasion and metastasis. To identify EMT-related breast cancer therapeutic targets and biomarkers, we have used two proteomic approaches to find proteins that change in abundance upon the induction of EMT by TGF-beta in two mouse mammary epithelial cell lines, NMuMG and BRI-JM01. RESULTS: Preliminary experiments based on two-dimensional electrophoresis of a hydrophobic cell fraction identified only 5 differentially expressed proteins from BRI-JM01 cells. Since 3 of these proteins were glycoproteins, we next used the lectin, wheat germ agglutinin (WGA), to enrich for glycoproteins, followed by relative quantification of tryptic peptides using a label-free LC-MS based method. Using these approaches, we identified several proteins that are modulated during the EMT process, including cell adhesion molecules (several members of the Integrin family, Fibronectin, Activated leukocyte cell adhesion molecule, and Neural cell adhesion molecule 1) and regulators of cellular signaling (Tumor-associated calcium signal transducer 2, Basigin). CONCLUSION: Interestingly, despite the fact that TGF-beta induces similar EMT phenotypes in NMuMG and BRI-JM01 cells, the proteomic results for the two cell lines showed only minimal overlap. These differences likely result in part from the conservative cut-off values used to define differentially-expressed proteins in these experiments. Alternatively, it is possible that the two cell lines may use different mechanisms to achieve an EMT transition. PMID- 19128515 TI - Genes involved in arsenic transformation and resistance associated with different levels of arsenic-contaminated soils. AB - BACKGROUND: Arsenic is known as a toxic metalloid, which primarily exists in inorganic form [As(III) and As(V)] and can be transformed by microbial redox processes in the natural environment. As(III) is much more toxic and mobile than As(V), hence microbial arsenic redox transformation has a major impact on arsenic toxicity and mobility which can greatly influence the human health. Our main purpose was to investigate the distribution and diversity of microbial arsenite resistant species in three different arsenic-contaminated soils, and further study the As(III) resistance levels and related functional genes of these species. RESULTS: A total of 58 arsenite-resistant bacteria were identified from soils with three different arsenic-contaminated levels. Highly arsenite-resistant bacteria (MIC > 20 mM) were only isolated from the highly arsenic-contaminated site and belonged to Acinetobacter, Agrobacterium, Arthrobacter, Comamonas, Rhodococcus, Stenotrophomonas and Pseudomonas. Five arsenite-oxidizing bacteria that belonged to Achromobacter, Agrobacterium and Pseudomonas were identified and displayed a higher average arsenite resistance level than the non-arsenite oxidizers. 5 aoxB genes encoding arsenite oxidase and 51 arsenite transporter genes [18 arsB, 12 ACR3(1) and 21 ACR3(2)] were successfully amplified from these strains using PCR with degenerate primers. The aoxB genes were specific for the arsenite-oxidizing bacteria. Strains containing both an arsenite oxidase gene (aoxB) and an arsenite transporter gene (ACR3 or arsB) displayed a higher average arsenite resistance level than those possessing an arsenite transporter gene only. Horizontal transfer of ACR3(2) and arsB appeared to have occurred in strains that were primarily isolated from the highly arsenic-contaminated soil. CONCLUSION: Soils with long-term arsenic contamination may result in the evolution of highly diverse arsenite-resistant bacteria and such diversity was probably caused in part by horizontal gene transfer events. Bacteria capable of both arsenite oxidation and arsenite efflux mechanisms had an elevated arsenite resistance level. PMID- 19128516 TI - A gene expression signature shared by human mature oocytes and embryonic stem cells. AB - BACKGROUND: The first week of human pre-embryo development is characterized by the induction of totipotency and then pluripotency. The understanding of this delicate process will have far reaching implication for in vitro fertilization and regenerative medicine. Human mature MII oocytes and embryonic stem (ES) cells are both able to achieve the feat of cell reprogramming towards pluripotency, either by somatic cell nuclear transfer or by cell fusion, respectively. Comparison of the transcriptome of these two cell types may highlight genes that are involved in pluripotency initiation. RESULTS: Based on a microarray compendium of 205 samples, we compared the gene expression profile of mature MII oocytes and human ES cells (hESC) to that of somatic tissues. We identified a common oocyte/hESC gene expression profile, which included a strong cell cycle signature, genes associated with pluripotency such as LIN28 and TDGF1, a large chromatin remodelling network (TOP2A, DNMT3B, JARID2, SMARCA5, CBX1, CBX5), 18 different zinc finger transcription factors, including ZNF84, and several still poorly annotated genes such as KLHL7, MRS2, or the Selenophosphate synthetase 1 (SEPHS1). Interestingly, a large set of genes was also found to code for proteins involved in the ubiquitination and proteasome pathway. Upon hESC differentiation into embryoid bodies, the transcription of this pathway declined. In vitro, we observed a selective sensitivity of hESC to the inhibition of the activity of the proteasome. CONCLUSION: These results shed light on the gene networks that are concurrently overexpressed by the two human cell types with somatic cell reprogramming properties. PMID- 19128517 TI - Usefulness and safety of 0.4% sodium hyaluronate solution as a submucosal fluid "cushion" for endoscopic resection of colorectal mucosal neoplasms: a prospective multi-center open-label trial. AB - BACKGROUND: Sodium hyaluronate (SH) solution has been used for submucosal injection in endoscopic resection to create a long-lasting submucosal fluid "cushion". Recently, we proved the usefulness and safety of 0.4% SH solution in endoscopic resection for gastric mucosal tumors. To evaluate the usefulness of 0.4% SH as a submucosal injection solution for colorectal endoscopic resection, we conducted an open-label clinical trial on six referral hospitals in Japan. METHODS: A prospective multi-center open-label study was designed. A total of 41 patients with 5-20 mm neoplastic lesions localized in the colorectal mucosa at six referral hospitals in Japan in a single year period from December 2002 to November 2003 were enrolled and underwent endoscopic resection with SH. The usefulness of 0.4% SH was assessed by the en bloc complete resection and the formation and maintenance of mucosal lesion-lifting during endoscopic resection. Safety was evaluated by analyzing adverse events during the study period. RESULTS: The usefulness rate was high (82.5%; 33/40). The following secondary outcome measures were noted: 1) steepness of mucosal lesion-lifting, 75.0% (30/40); 2) intraoperative complications, 10.0% (4/40); 3) time required for mucosal resection, 6.7 min; 4) volume of submucosal injection, 6.8 mL and 5) ease of mucosal resection, 87.5% (35/40). Two adverse events of bleeding potentially related to 0.4% SH were reported. CONCLUSION: Using 0.4% SH solution enabled sufficient lifting of a colorectal intramucosal lesion during endoscopic resection, reducing the need for additional injections and the risk of perforation. Therefore, 0.4% SH may contribute to the reduction of complications and serve as a promising submucosal injection solution due to its potentially superior safety in comparison to normal saline solution. PMID- 19128518 TI - Phlebotomus (Euphlebotomus) barguesae n. sp. from Thailand (Diptera - Psychodidae). AB - BACKGROUND: A few studies have been carried out on the Phlebotomine sandflies from Thailand. Within the Phlebotomine sandflies, the genus Phlebotomus Rondani & Berte, 1840 contains the vectors of leishmaniases in Europe, Africa and Asia. It includes several subgenera. Among them the subgenus Euphlebotomus Theodor, 1948 contains at the present time 12 taxa. The type-species of this subgenus is P. argentipes Annandale & Brunetti, 1908, the vector of Leishmania donovani (Laveran & Mesnil, 1903) in India. RESULTS: A new species of sandfly, P. barguesae n. sp. is described from limestone caves in Thailand. The male-female gathering in the same species is based on ecological, morphological and molecular criteria (homology of mtDNA cytochrome c oxidase I sequences). The inclusion of P. barguesae n. sp. in the subgenus Euphlebotomus is justified on the basis of characters of the male genitalia (five spines on the style, bifurcated paramere, and no basal lobe on the coxite) and of female pharyngeal armature (two kinds of teeth). It well differenciated from another sympatric species: P. mascomai. CONCLUSION: The new species described in the present study has smooth spermathecae. This original morphology opens a discussion on the heterogeneity of this subgenus. PMID- 19128519 TI - Leishmaniasis presenting to the otolaryngologist: a rare but important cause of persistent hoarseness. AB - OBJECTIVE: We report a rare UK case of laryngeal leishmaniasis, mimicking laryngeal candidiasis, associated with long term steroid inhaler use. METHODS: Case report and review of the world literature concerning leishmaniasis. RESULTS: Laryngeal leishmaniasis is a protozoal infection which is rare in the Western world. It is becoming more common, however, with increased foreign travel. The disease can be difficult to diagnose histologically, and diagnosis is often delayed because of its rarity. It can mimic malignant laryngeal disease, and patients may therefore be subjected to significant and inappropriate treatment interventions. CONCLUSIONS: A diagnosis of leishmaniasis should be considered if initial treatment for persistent hoarseness is ineffective, particularly in a patient who is at low risk of malignancy. PMID- 19128520 TI - Laryngolith. AB - OBJECTIVE: To report an extremely rare and interesting case of a woman who developed a laryngeal stone. CASE REPORT: A 44-year-old woman was referred to our ENT clinic complaining of dyspnoea and stridor. She had been treated for chest problems for approximately 30 years. On examination, she had a large, spiky, subglottic lesion compromising the airway. A laryngeal computed tomography scan and histopathological and biochemical examination showed the lesion to be a calcium phosphate stone. Review of the literature from the past 50 years, using several search engines, revealed no similar cases. CONCLUSION: To the author's knowledge, this is a unique case of a stone that developed in the subglottis. This rare entity was the cause of long-standing chest problems unresponsive to medical treatment. The patient recovered completely from all her symptoms after removal of the laryngolith. PMID- 19128521 TI - Does Young's syndrome exist? AB - BACKGROUND AND METHODS: Young's syndrome describes a combination of male infertility, azoospermia, bronchiectasis and sinusitis. Although Young's syndrome is a well accepted disorder within the realms of infertility medicine, it is also accepted as being a potential cause of sino-nasal disease which is rarely seen by otolaryngologists. However, the significance of the sinus component within this triad is not fully understood. To gain further insight into the relationship of sinusitis with Young's syndrome, we reviewed all of the currently available published literature. RESULTS: Within the reviewed literature, the diagnosis of sinusitis in Young's syndrome was crude and poorly defined; there was little emphasis on sinus disease in most publications. CONCLUSIONS: The prevalence of Young's syndrome is reported to be declining, and the level of evidence regarding sinus disease within this syndrome is limited to case series only. There is, in fact, little evidence to support Young's syndrome being a significant aetiological factor for sinus disease, nor indeed to support the existence of Young's syndrome as an entity in its own right. The only documented aetiological factor is mercury exposure in childhood, an event that is seldom currently encountered; this would support our theory of the extinction of the condition. As an incidental finding, we found that the term Young's syndrome refers to two different medical conditions. PMID- 19128522 TI - Mapping surgical coordinates of the sphenopalatine foramen: surgical navigation study. AB - OBJECTIVES: To identify measurements that may help intra-operative localisation of the sphenopalatine foramen. DESIGN: The study used three-dimensional surgical navigation software to study radiological anatomy, in order to define the distances and angulations between identifiable bony landmarks and the sphenopalatine foramen. RESULTS: The distance from the anterior nasal spine to the sphenopalatine foramen was 59 mm (+/-4 mm; inter-observer variation = 0.866; intra-observer variation = 0.822). The distance from the piriform aperture to the sphenopalatine foramen was 48 mm (+/-4 mm; inter-observer variation = 0.828; intra-observer variation = 0.779). The angle of elevation from the nasal floor to the sphenopalatine foramen was 22 degrees (+/-3 degrees; inter-observer variation = 0.441; intra-observer variation = 0.499). CONCLUSIONS: The sphenopalatine foramen is consistently identifiable on three-dimensional, reconstructed computed tomography scans. Repeatable measurements were obtained. The centre point of the foramen lies 59 mm from the anterior nasal spine at 22 degrees elevation above the plane of the hard palate and 48 mm from the piriform aperture. We discuss how these data could be used to facilitate intra-operative location of the sphenopalatine foramen in difficult cases. PMID- 19128523 TI - Cognitive sequelae of blast-related versus other mechanisms of brain trauma. AB - The use of improvised explosive devices has become the hallmark of modern warfare and has resulted in an ever-increasing number of blast-related traumatic brain injuries (TBIs). Despite this fact, very little is actually known about the cognitive sequelae of blast-related TBIs. The purpose of the current study was to compare patterns of performance on neuropsychological measures in subjects who have sustained TBIs as a result of blast (or explosion) with those who have sustained TBIs from non-blast or blunt force trauma (motor vehicle accident, fall, assault, etc.). Participants were categorized as blast-related TBI or non blast-related TBI and according to severity of injury (mild or moderate-to severe). No main effects were observed in analysis of covariance between blast related TBI participants and non-blast-related TBI participants across any of the neuropsychological variables, although an interaction was observed on a visual memory test showing stronger performance for mild blast-related and poorer performance for moderate-to-severe blast-related participants compared with both non-blast groups. Overall, the results do not provide any strong evidence that blast is categorically different from other TBI mechanisms, at least with regard to cognitive sequelae on select measures. Additional findings included a marginally increased incidence of reported posttraumatic stress disorder symptoms among blast-injured participants. (JINS, 2009, 15, 1-8.). PMID- 19128525 TI - Dimensions of postconcussive symptoms in children with mild traumatic brain injuries. AB - The dimensions of postconcussive symptoms (PCS) were examined in a prospective, longitudinal study of 186 8 to 15 year old children with mild traumatic brain injuries (TBI). Parents and children completed a 50-item questionnaire within 2 weeks of injury and again at 3 months after injury, rating the frequency of PCS on a 4-point scale. Common factor analysis with target rotation was used to rotate the ratings to four hypothesized dimensions, representing cognitive, somatic, emotional, and behavioral symptoms. The rotated factor matrix for baseline parent ratings was consistent with the target matrix. The rotated matrix for baseline child ratings was consistent with the target matrix for cognitive and somatic symptoms but not for emotional and behavioral symptoms. The rotated matrices for ratings obtained 3 months after injury were largely consistent with the target matrix derived from analyses of baseline ratings, except that parent ratings of behavioral symptoms did not cluster as before. Parent and child ratings of PCS following mild TBI yield consistent factors reflecting cognitive and somatic symptom dimensions, but dimensions of emotional and behavioral symptoms are less robust across time and raters. (JINS, 2009, 15, 19-30.). PMID- 19128524 TI - Semantic knowledge for famous names in mild cognitive impairment. AB - Person identification represents a unique category of semantic knowledge that is commonly impaired in Alzheimer's disease (AD), but has received relatively little investigation in patients with mild cognitive impairment (MCI). The current study examined the retrieval of semantic knowledge for famous names from three time epochs (recent, remote, and enduring) in two participant groups: 23 amnestic MCI (aMCI) patients and 23 healthy elderly controls. The aMCI group was less accurate and produced less semantic knowledge than controls for famous names. Names from the enduring period were recognized faster than both recent and remote names in both groups, and remote names were recognized more quickly than recent names. Episodic memory performance was correlated with greater semantic knowledge particularly for recent names. We suggest that the anterograde memory deficits in the aMCI group interferes with learning of recent famous names and as a result produces difficulties with updating and integrating new semantic information with previously stored information. The implications of these findings for characterizing semantic memory deficits in MCI are discussed. (JINS, 2009, 15, 9 18.). PMID- 19128526 TI - Neuroimaging correlates of parent ratings of working memory in typically developing children. AB - The purpose of the present study was to investigate construct validity of parent ratings of working memory in children, using a multi-trait/multi-method design including neuroimaging, rating scales, and performance-based measures. Thirty five typically developing children completed performance-based tests of working memory and nonexecutive function (EF) skills, received volumetric MRI, and were rated by parents on both EF-specific and broad behavior rating scales. After controlling for total cerebral volume and age, parent ratings of working memory were significantly correlated with frontal gray, but not temporal, parietal, or occipital gray, or any lobar white matter volumes. Performance-based measures of working memory were also moderately correlated with frontal lobe gray matter volume; however, non-EF parent ratings and non-EF performance-based measures were not correlated with frontal lobe volumes. Results provide preliminary support for the convergent and discriminant validity of parent ratings of working memory, and emphasize their utility in exploring brain-behavior relationships in children. Rating scales that directly examine EF skills may potentially have ecological validity, not only for "everyday" function, but also as correlates of brain volume. (JINS, 2009, 15, 31-41.). PMID- 19128527 TI - Timing is everything: antiretroviral nonadherence is associated with impairment in time-based prospective memory. AB - Nonadherence to combination antiretroviral (ARV) therapies (cART) is highly prevalent and significantly increases the risk of adverse human immunodeficiency virus (HIV) disease outcomes. The current study evaluated the hypothesis that prospective memory-a dissociable aspect of episodic memory describing the ability to execute a future intention-plays an important role in successful cART adherence. Seventy-nine individuals with HIV infection who were prescribed at least one ARV medication underwent a comprehensive neuropsychological and neuromedical evaluation prior to completing a 1-month observation of their cART adherence as measured by electronic medication monitoring. Nonadherent individuals (n = 31) demonstrated significantly poorer prospective memory functioning as compared to adherent persons (n = 48), particularly on an index of time-based ProM (i.e., elevated loss of time errors). Deficits in time-based prospective memory were independently predictive of cART nonadherence, even after considering the possible influence of established predictors of adherence, such as general cognitive impairment (e.g., retrospective learning and memory) and psychiatric comorbidity (e.g., depression). These findings extend a nascent literature showing that impairment in time-based prospective memory significantly increases the risk of medication nonadherence and therefore may guide the development of novel strategies for intervention. (JINS, 2009, 15, 42-52.). PMID- 19128528 TI - Negative affect predicts subsequent cognitive change in multiple sclerosis. AB - Baseline predictors of cognitive change were explored in a sample of persons with multiple sclerosis (MS). Potential predictors included demographic features, baseline clinical characteristics, and psychological state. Participants were 38 individuals diagnosed with either relapsing remitting or secondary progressive MS who did not meet criteria for a current major depressive episode. Subjects were tested at baseline and approximately 1 year in an ongoing longitudinal study of cognition in MS. Participants completed neuropsychological tests sensitive to impairment in MS. They also completed self-report measures of depression, anxiety, fatigue, apathy, and positive and negative affect. Baseline measures of negative affect (e.g., depressed mood, state anxiety, and negative affective state) consistently predicted cognitive change over the course of the study. Higher baseline levels of negative affect were associated with greater relative declines in cognitive performance. This longitudinal relation occurred in the absence of a cross-sectional relation between negative affect and overall cognition. High baseline negative affect particularly predicted a relative decline in episodic memory for newly learned verbal and visuospatial information. The negative affect measures were unique in their predictive value among all the baseline measures assessed. (JINS, 2009, 15, 53-61.). PMID- 19128529 TI - Emotional incontinence and executive function in ischemic stroke: a case controlled study. AB - Frontal and basal ganglia infarcts and executive dysfunction are thought to be involved in the pathophysiology of poststroke emotional incontinence (PSEI). The study examined whether patients with PSEI have more frontal and/or basal ganglia infarcts and impairment in executive function. A total of 516 Chinese patients with acute ischemic stroke consecutively admitted to the acute stroke unit of a university-affiliated regional hospital in Hong Kong were screened for PSEI 3 months after the index stroke. According to Kim's criteria, 39 (7.6%) had PSEI. Thirty-nine stroke patients without PSEI served as matched control group. The PSEI group had significantly more frontal and/or basal ganglia infarcts, had lower Chinese Frontal Assessment Battery scores, required more time to complete the Stroop Test, and made more omission and commission errors in the Go-NoGo test. There was no significant correlation between frontal or basal ganglia infarcts and executive function. The correlation between frontal infarct and severity of PSEI was .420. Further follow-up and functional imaging studies are warranted to explore the relationship between PSEI, brain infarcts, and executive dysfunction. (JINS, 2009, 15, 62-68.). PMID- 19128531 TI - Proactive and retroactive interference in young adults, healthy older adults, and older adults with amnestic mild cognitive impairment. AB - This study investigated memory interference in amnestic mild cognitive impairment (aMCI) and normal aging. Participants were 27 young adults, 44 healthy older adults, and 15 older adults with aMCI. Memory interference was examined on the California Verbal Learning Test (CVLT) and on a modified AB-AC paradigm. Despite significant differences in memory performance on the CVLT, interference measures on this test did not distinguish individuals with aMCI and healthy older adults. The AB-AC task involved first learning a list (AB list) of 12 semantically related word pairs (e.g., knee-bone). Twenty minutes later, a second list (AC list) was learned in which the same stimulus words were paired with new response words (e.g., knee-bend). Both lists were repeated until 100% accurate recall was achieved. Finally, participants recalled the first (AB) list. Proactive interference (PI) was greater among older than younger adults, and greater still among individuals with aMCI, but the older and aMCI groups showed similar levels of retroactive interference. This study suggests that PI contributes to the memory deficits seen in aMCI and that tests sensitive to PI may assist in the early identification of aMCI. Memory interventions aimed at alleviating PI may improve memory functioning in individuals with aMCI. (JINS, 2009, 15, 83-93.). PMID- 19128530 TI - The cognitive effects of hepatitis C in the presence and absence of a history of substance use disorder. AB - The aim of the study was to determine whether infection with the hepatitis C virus (HCV) is associated with cognitive impairment beyond the effects of prevalent comorbidities and a history of substance use disorder (SUD). Adult veterans were recruited from the Portland Veterans Affairs Medical Center into three groups: (1) HCV+/SUD+ (n = 39), (2) HCV+/SUD- (n = 24), and (3) HCV-/SUD- (n = 56). SUD+ participants were in remission for > or =90 days, while SUD- participants had no history of SUD. Groups did not significantly differ in terms of rates of psychiatric or medical comorbidities. Procedures included clinical interviews, medical record reviews, and neuropsychological testing. Significant group differences were found in the domains of Verbal Memory, Auditory Attention, Speeded Visual Information Processing, and Reasoning/Mental Flexibility (p 0.05). DISCUSSION: Educational programs and enrollment of more female orthopaedic surgeons may enhance ECG skills in the orthopaedic surgical community. PMID- 19128557 TI - [Hearing disorders and rock music]. AB - Only few studies have investigated the frequency of hearing disorders in rock musicians. Performing rock music is apparently associated with a hearing loss in a fraction of musicians. Tinnitus and hyperacusis are more common among rock musicians than among the background population. It seems as if some sort of resistance against further hearing loss is developed over time. The use of ear protection devices have not been studied systematically but appears to be associated with diminished hearing loss. PMID- 19128558 TI - [The effect of fasting on the surgical skills of surgeons]. AB - There have been worries of a potential negative effect of the fasting practice on surgeon's skills. We set out to examine the effect of fasting for a single surgeon during the fast and after the fast was broken. We examined the skills of the surgeon in a previously developed model for training in laparoscopic surgery. Foetor ex ore was also examined during and after the fast. Fasting did not affect the surgical skills, but resulted in increased foetor ex ore. Abstaining from eating and drinking during the daytime does probably not negatively affect surgical skills, but increases foetor ex ore. PMID- 19128559 TI - [Our children stuff themselves with medications--does it matter?]. PMID- 19128560 TI - [Children's and adolescent's use of medicine for aches and psychological problems: secular trends from 1988 to 2006]. AB - INTRODUCTION: Medicine use for aches and psychological problems is common among adolescents. Medicines are toxic and may have harmful side effects. It is therefore important to study change over time and patterns of medicine use. The objective of this paper is to describe self-reported medicine use for headaches, stomach-aches, difficulties in falling asleep, and nervousness among 11-, 13-, and 15-year-old boys and girls from 1968 to 2006. MATERIAL AND METHODS: The data material is 6 comparable and representative cross-sectional studies of 11-, 13-, and 15-year-olds from 1988, 1991, 1994, 1998, 2002, and 2006, total n = 23,871. Data were collected by a validated self-report method for medicine use and prevalence of the corresponding complaints. RESULTS: The proportion of students who used medicine for headaches within the previous month increased from 24.6% in 1988 to 42.0% in 2002 and 41.5% in 2006. The proportion who used medicine for stomach-aches increased from 7.5% in 1988 to 16.2% in 2002 and 15.2% in 2006. The proportion who used medicine for difficulties in falling asleep increased from 2.2% to 5.1%, and the proportion who used medicine for nervousness increased from 0.7% to 3.8%. There were increasing proportions of users in most sex- and age specific subgroups. In the same period, the proportion who reported headaches and stomach-aches within the previous month decreased. The proportion who reported nervousness and difficulties in falling asleep did not change much in this period. CONCLUSION: There was a significant increase in 11-, 13-, and 15-year-old student's use of medicine for aches and psychological problems from 1988 to 2006. In the same period, there was a decrease in the prevalence of students who reported pains monthly. PMID- 19128561 TI - [Congenital diaphragmatic hernia in Western Denmark]. AB - INTRODUCTION: The aim was to clarify the mortality rate and results of surgical treatment in children with congenital diaphragmatic hernia (CDH) born in Western Denmark. MATERIAL AND METHODS: A retrospective cohort study of all live-born infants with CDH born in Western Denmark between 1995 and 2005. The cohort enrolled 82 children, which was divided into two groups, before and after 1998, when the multidisciplinary treatment centre at Odense University Hospital (OUH) was established. RESULTS: The overall mortality between 1998 and 2005 was 23%, which was significantly lower than between 1995 and 1998, when it was 67%. The frequency of vaginal delivery, treatment with high-frequency oscillatory ventilation (HFO) and inotropic medications was higher in the second period, but the difference was not statistically significant. There was no access to extracorporal membranous oxygenation (ECMO). Postoperative complications were registered in half of the children. Three children were re-operated due to recurrence of the hernia. CONCLUSION: The overall mortality rate in live-born children with CDH was 23% in the setting of a multidisciplinary treatment centre, which was similar to that reported by other centres. In cases with prenatally diagnosed CDH, delivery in a multidisciplinary treatment centre is recommended. PMID- 19128562 TI - [Diaphragmatic hernia--admitted under the gallstone diagnosis]. AB - A 34-year old woman was admitted with pain in the upper right abdomen. The tentative diagnosis was gall stones, but none of the paraclinical results supported this diagnosis. A chest x-ray showed a completely white right lung. A later computertomography showed a colon-loop in the thorax and haemo /pneumothorax. The patient's condition deteriorated, a life-threatening state of sepsis developed and she was transferred to a surgery facility where an incarcerated congenital diaphragmatic hernia was established. After surgery she was discharged with a temporary ostomy, but otherwise feeling well. PMID- 19128563 TI - [Computer tomographic diagnosis of an internal hernia]. AB - A case is presented in which a transmesenteric hernia developed four weeks after transperitoneal laparoscopic left nephrectomy. The diagnosis of an internal hernia is best established by computer tomography; however, the signs have low sensitivity, and on suspicion of an internal hernia, early laparoscopy or laparotomy is recommended. PMID- 19128564 TI - [Diaphragmatic hernia presenting with colonic strangulation]. AB - Diaphragmatic hernia may be asymptomatic and may have a delayed clinical manifestation. We describe a 78-year-old man who developed colonic obstruction as a complication to a diaphragmatic hernia. Unlike most diaphragmatic hernias, this case was not associated with a trauma. Normally, the diagnosis is made clinically by means of thoracic X-ray, computer tomography or magnetic resonance imagining of the abdomen. In this patient, barium enema showed a tumor-like stenosis of the left colonic flexure, but intraoperative findings demonstrated a defect in the left diaphragm with herniation of the left colonic flexure and the greater omentum. PMID- 19128565 TI - [Long-term prognosis for developmental language disorder]. AB - INTRODUCTION: The purpose of the study was to examine the effect of neurologic and psychiatric comorbidity on the prognosis of developmental language disorder. MATERIAL AND METHODS: A total of 470 pre-school children (4-7 years) with language learning impairment were investigated in the period 1969-1976 at the Institute for Language Impairment in Risskov (SIT) during a 3-6 week observation period. Deaf children were excluded. Questionnaires were sent to the 470 participants in 2006. They were then 36-45 years of age. RESULTS: A total of 200 responses were received. The answer group and the non-responding groups were equal with regard to all parameters investigated in childhood. For 41% a pure hereditary aetiology was present, while 58% had neurological and 51% psychiatric comorbidity. In all, 77% of the children developed reading difficulties during the first school years, while 54% had reading difficulties at the follow-up. 40% started an education after leaving school, while 29% finished their training and acquired work corresponding to their attained level of training. At the follow up, 35% were self-supporting while 65% received a state pension. Neurological comorbidity was decisive for the prognosis. DISCUSSION: Neurological and psychiatric comorbidity is frequent in children with medium and severe language learning impairment and are decisive for the prognosis. Examination of children suspected of brain dysfunction by a paediatric neurologist and paediatric neuropsychologist is necessary to discover the cognitive deficits which require special habilitating treatment. PMID- 19128566 TI - [Endovascular treatment of dysfunctional haemodialysis fistulas]. AB - INTRODUCTION: A native AV-fistula (AVF) is the first choice of access for haemodialysis. An important complication is thrombosis, which typically results from an underlying stenosis. In 2001, we introduced a surveillance program measuring the access flow (Qa). At the same time percutaneous transluminal angioplasty (PTA) was introduced as the first choice of treatment. MATERIAL AND METHODS: Qa was conducted in a routine monitoring programme. Flow rates below a preset value indicated patient referral for angiography, and if the angiography revealed a significant stenosis, PTA was performed. In the period from August 2001 to March 2004, 71 angioplasties were performed in 52 patients. RESULTS: The technical success rate was 66/71 (93%). Clinically significant complications occurred at a rate of 3/71 (4%). The 2-year primary patency rate was 36% and the primary assisted patency rate was 67%. CONCLUSION: PTA in native haemodialysis fistulas was implemented. The rates of technical success, complications and patency were comparable with those recorded in the literature. PMID- 19128568 TI - [Bisphosphonate-associated osteonecrosis of the jaw in patients with multiple myeloma]. AB - Bisphosphonate-associated osteonecrosis of the jaw (BON) is mainly observed in patients with multiple myeloma, and to a lesser extent in breast and prostate cancer patients receiving intravenous treatment with potent bisphosphonates. The incidence of BON increases with the duration of bisphosphonate therapy and with the potency of the used bisphosphonate. BON usually develops after tooth extraction or other oral surgery, and has proven difficult to treat. Optimal dental hygiene should be ensured prior to treatment initiation where possible, and once bisphosphonate treatment is instituted, oral surgery should be avoided if possible. PMID- 19128567 TI - [Novel tonsillectomy technique]. AB - INTRODUCTION: Coblation (Co) is a relatively new technique, which may offer a better postoperative course after tonsillectomy than traditional techniques. The purpose of this study was to determine whether Co outperforms classic dissection. Coblation uses radiofrequency energy to excite the electrolytes in a saline solution, creating a precisely focused plasma. The plasma's energized particles have sufficient energy to break molecular bonds within tissue, causing the tissue to dissolve at temperatures between 40 degrees C and 70 degrees C. MATERIAL AND METHODS: A case-control study was conducted in which 26 patients undergoing Co tonsillectomy were matched concerning gender, age and the surgeon's charge with 26 patients undergoing classic tonsillectomy. Furthermore, to compare the rates of postoperative haemorrhage, we reviewed the health records of all patients undergoing Co (60 patients) and routine dissection tonsillectomy (403) at Roskilde County Hospital. RESULTS: Intraoperative bleeding was significantly lower in the Co group (median (MD) 5.0 ml; range (R) 1-32 ml) than in the control group (MD 10.0 ml; R 5-200 ml) (p < 0.0001). The median operation time was also significantly shorter for the Co group (23 min.; R 13-40 min. vs. 32 min.; R 21 63 min.; p = 0.002). Postoperatively, there was no significant difference between the two groups regarding pain scores, analgesics intake, time to return to work and normal diet or weight changes. Neither the primary (0% vs. 2%, p = 0.60) nor the secondary (3.3% vs. 2.5%, p = 0,66) haemorrhage rate was significantly different between Co and traditional tonsillectomy. CONCLUSION: The intraoperative bleeding was lower and the surgery times were shorter in the Co group compared with classic dissection. There was no difference in the postoperative haemorrhage rate between the two tonsillectomy methods. PMID- 19128569 TI - [Peripartum cardiomyopathy]. AB - Peripartum cardiomyopathy is a rare and potentially fatal disease with peripartum heart failure in previously healthy women. A brief review of the literature is provided with emphasis on recent data indicating that an antiangiogenic cleavage product of prolactin contributes to the molecular mechanisms underlying peripartum cardiomyopathy, and that blocking the release of prolactin with bromocriptine can ameliorate the condition. PMID- 19128570 TI - [Reconstructive techniques after rectal resection for rectal cancer. A survey of a Cochrane review]. PMID- 19128572 TI - [Subdural haematoma from aneurysm without concurrent subarachnoid haemorrhage]. AB - A case of acute subdural haematoma from an intracranial aneurysm is presented. Although the patient presented with isolated subdural haematoma, the clinical signs were consistent with the classical signs of subarachnoid haemorrhage including thunderclap headache. An aneurysm of the anterior cerebral artery was the origin of the bleeding, and no subarachnoid blood was identified during operation. Rupture of a sacculate aneurysm should be suspected in patients with non-traumatic acute subdural haematoma. PMID- 19128571 TI - [Small-bowel ileus caused by a gold filling]. AB - A 68-years-old man was admitted to hospital with signs of intestinal obstruction. Computed tomography showed a foreign body, and the patient underwent laparotomy which revealed a gold filling in the jejunum. To our knowledge, this is the first documented case of a gold filling causing intestinal obstruction. PMID- 19128573 TI - [Life-threatening subdural haematoma in young man]. AB - Pre-existence of an arachnoid cyst has previously been suggested to increase the risk of developing a subdural haematoma (SDH). In this case a young man was submitted acutely due to decreasing level of consciousness. Four weeks previously he had been hit by a football and suffered minor and transcient headache. Within minutes after submission, a further decrease in consciousness according to the Glasgow comascore was observed. Acute computed tomography showed SDH and an arachnoid cyst. Surgery was performed immediate and hours later he was improving. PMID- 19128575 TI - Neck dissection following concurrent chemoradiation for advanced head and neck carcinoma: pathologic findings and complications. AB - OBJECTIVE: To assess the rate of pathologic residual neck disease and complication rates following selective neck dissection in patients with advanced head and neck carcinoma after concurrent chemotherapy (CT) and radiation therapy (RT). PATIENTS AND METHODS: Two hundred sixty-four patients with advanced-stage head and neck carcinoma underwent concurrent CT and RT from 1998 to 2004 at Notre Dame Hospital. We describe the outcome of 32 of these patients who underwent neck dissections at our institution. The median age was 56 years (35-73 years). The primary sites were the oropharynx (n = 25), followed by the larynx (n = 3), hypopharynx (n = 1), and oral cavity (n = 1), and there were two unknown primary sites. The initial neck stages were 2 N1, 19 N2, and 11 N3. The known primary sites were staged 2 T1, 10 T2, 8 T3, 10 T4, and 2 Tx. All patients received conventional RT to a dose of 70 Gy in 35 fractions concurrent with a platinum based CT regimen RESULTS: The median follow-up was 20 months. The mean operative blood loss for neck dissections alone was 170 mL (SD = 158.3 mL), the mean operative time was 128 minutes (SD = 41.6 minutes), and the mean hospital stay was 3.6 days (SD = 1.7 days). There were no major surgical complications. Five minor complications were noted in four patients (12.5%). Twelve patients (38%) had pathologic evidence of persistent disease (1 of 2 N1, 7 of 19 N2, and 4 of 11 N3). CONCLUSIONS: Cervical neck dissection is a safe procedure following concurrent CT and RT and is still indicated if there is clinical or radiologic evidence of disease persistence in the neck. We experienced no major surgical complications and few minor complications. The pathologic residual rate was 38%. PMID- 19128576 TI - Evaluation of the frequency selectivity of contralateral acoustic stimulation on the active mechanisms of the organ of corti by analyzing the changes in the amplitude of transitory evoked otoacoustic emissions and distortion products. AB - INTRODUCTION: The central nervous system can regulate the input of acoustic information to the cochlea by means of the olivocochlear efferent system's action on the organ of Corti. The aim of this study was to determine whether the suppressor effect of contralateral acoustic stimulation shows frequency selectivity in active cochlear mechanisms by recording transient evoked otoacoustic emissions (TEOAEs) and distortion product otoacoustic emissions (DPOAEs). MATERIAL AND METHOD: We conducted a prospective study of 56 subjects with normal-hearing ears aged 20 to 22 years (mean 5 20.9 years); 50% were male and 50% were female. We studied the amplitude of each TEOAE and DPOAE frequency band before and after contralateral acoustic stimulation with broadband white noise, 0 to 20,000 Hz frequency range, and pure tones of 700, 1000, 1500, 2000, 3000, 4000, 5000, and 6000 Hz, at an intensity of 60 dB HL. RESULTS: TEOAE recording amplitudes decreased 84% after contralateral acoustic stimulation. White noise and pure tones of 1000, 1500, and 2000 Hz had the greatest suppressor effects on the TEOAEs. The suppressor effect was higher in 1000 to 4000 Hz frequency bands, with a statistically significant decrease in amplitudes of 0.5 to 2.5 dB. Distortion product amplitude decreased 75%. Stimulation with white noise and pure-tone contralateral stimulation at 1000 and 1500 Hz showed the highest decrease in DPOAE amplitude. Suppression was concentrated in DPOAEs obtained with F2 of 1500 and 2000 Hz. CONCLUSION: Contralateral acoustic stimulation causes selective frequency modulation of the cochlear micromechanisms, which can be assessed by recording TEOAEs and DPOAEs. PMID- 19128577 TI - Microdebrider for reduction of inferior turbinate: evaluation of effectiveness by computed tomography. AB - OBJECTIVE: The microdebrider has been widely used in rhinologic surgery in the last few years, and there are reports on its use for inferior turbinate reduction. Computed tomography (CT) is commonly employed to show the reduction in the inferior turbinate after surgery; however, measurements on the scans are rarely made. DESIGN: A prospective study. SETTING: A tertiary research and training hospital. METHODS: We investigated the effectiveness of microdebrider reduction in inferior turbinate hypertrophy in 12 turbinates of 9 patients. MAIN OUTCOME MEASURES: Mean bone thicknesses were measured in CT in the anterior and middle portions of the inferior turbinates as an objective parameter. Preoperative and postoperative visual analogue scale scores for symptoms and endoscopic grading of the inferior turbinates were also used. RESULTS: In the postoperative period, visual analogue scale scores for nasal obstruction decreased significantly. Endoscopic grading revealed that the inferior turbinates were significantly smaller in the postoperative period. Postoperative mean bone thicknesses measured in CT in the anterior and the middle portions of the inferior turbinates were significantly smaller than the preoperative ones; however, there was no significant difference between the measurements of the medial and lateral soft tissue thicknesses. CONCLUSIONS: We suggest microdebrider turbinate surgery as a reliable alternative method for inferior turbinate reduction, especially in case of hypertrophy of the turbinate bone. Measurement of the inferior turbinate size in paranasal CT is a useful objective method for evaluation of the inferior turbinate hypertrophy and the assessment of the surgical outcomes. PMID- 19128578 TI - Prevalence of adenoid hypertrophy in adults with nasal obstruction. AB - OBJECTIVE: To assess the prevalence of adenoid hypertrophy in adults presenting with nasal obstruction. DESIGN: A prospective study was conducted. Descriptive analysis was used for statistical analysis. SETTING: Fifty-five patients above the age of 17 years who presented with nasal obstruction were enrolled in the study. Forty-nine patients with no history of nasal obstruction were matched according to age and gender as controls. Allergy, pattern of nasal obstruction (continuous vs intermittent and unilateral vs bilateral), and symptoms such as nasal discharge, postnasal drip, headache, and facial pain were investigated in relation to the presence or absence of adenoid hypertrophy. Nasal findings such as the presence of intranasal polyps and potency of the osteomeatal complex were also reviewed. RESULTS: The overall prevalence of adenoid hypertrophy in adults with nasal obstruction approached 63.6% in patients with nasal obstruction and 55.1% in the control group (p = .007). CONCLUSION: Adenoid hypertrophy is often underestimated in adults with nasal obstruction. The presence of purulent nasal discharge should stimulate the caring physician to do nasal endoscopy for proper diagnosis. PMID- 19128579 TI - Image-guided endoscopic transsphenoidal removal of pituitary tumours. AB - BACKGROUND: With continuing advancements in minimally invasive endoscopic nasal and sinus surgical techniques, image-guided endoscopic transsphenoidal approaches to the pituitary gland and anterior skull base have progressively become more popular. METHODS: Clinical records were reviewed retrospectively from 2001 to 2006. Twenty-nine consecutive patients with pituitary tumours underwent transnasal endoscopic surgery. RESULTS: We outlined our endoscopic surgical technique with the use of image guidance. The duration of surgery, complications, and hospital stay were also reviewed. CONCLUSIONS: Image guidance can be applied during transnasal transsphenoidal endoscopic surgery and requires a minimal amount of time. It can reduce morbidity and allow for a more complete and safer operation with minimal postoperative discomfort. It can significantly reduce postoperative hospital stay, resulting in a reduced cost to the medical system. PMID- 19128580 TI - Endoscopic management of acquired cholesteatoma: our experience. AB - OBJECTIVES: The purpose of this study was to examine the utility of using an endoscope in cholesteatoma surgery and to demonstrate how it allows a reduction in the incidence of residual disease. MATERIAL AND METHODS: Thirty-two ears with acquired cholesteatoma (primary) were resected. Twenty cases were resected using a canal wall up (CWU) technique and six cases using a canal wall down (CWD) technique, and in six cases, a transcanal tympanotomy-atticotomy was performed. All of the patients in our study group underwent explorative and operative endoscopic ear surgery complementary to use of the operating microscope to uncover and remove residual cholesteatoma. RESULTS: In the primary surgery after completion of microscopic cleaning, the overall incidence of intraoperative residual disease detected with the endoscope was 37.5%. The sinus tympani was the most common site of intraoperative residual disease, followed by the anterior epitympanic recess and protympanum. Of the 20 CWU cases, 12 second-look endoscopies were performed. Two residuals were identified, both in the sinus tympani. No significant complications were associated with the 32 endoscopic procedures. CONCLUSION: The endoscope allowed a better understanding of cholesteatoma and improved eradication of residual disease from hidden areas such as the anterior epitympanic recess, retrotympanum, and hypotympanum not yet controllable by the operating microscope. PMID- 19128581 TI - Impact of cochlear implantation on speech understanding, depression, and loneliness in the elderly. AB - OBJECTIVE: To compare speech understanding ability, the level of depression, and the degree of loneliness experienced by elderly cochlear implant (CI) users (>or= 70 years), adult CI users (or= 70 years). DESIGN: Clinical study. SETTING: Tertiary academic neuro otologic and audiologic centre. METHODS: Three groups of patients were enrolled: (1) nine unilateral CI users 70 years or older at the time of implantation (mean 77.7 years), (2) eight unilateral CI users 60 years or younger at the time of implantation (mean 51.1 years), and (3) nine bilateral HA users 70 years or older (mean 77.5 years). Subjects underwent speech perception testing and completed two scales regarding their perceived levels of depression (pre-/post-CI) and feelings of loneliness (pre-/post-CI). MAIN OUTCOME MEASURE: Speech understanding scores in quiet and in noise and quality of life indicators (UCLA Loneliness Questionnaire, Geriatric Depression Screening Scale). RESULTS: There were no perioperative complications. No significant differences in speech understanding ability in quiet or in noise between elderly and younger CI patients were observed (p < .05). Cochlear implantation decreased perceived depression in elderly recipients and loneliness in both elderly and younger recipients. Finally, elderly CI users were no more depressed or lonely than their age-matched peers with mild-to-moderate hearing loss who use HAs. CONCLUSIONS: Cochlear implantation in elderly patients results in speech perception abilities comparable to those of younger CI recipients, as well as measurable improvements in depression and loneliness. PMID- 19128582 TI - [Personal and social identity transformations that occur over time among patients with total laryngectomy]. AB - OBJECTIVE: To examine personal and social identity transformations that occur over time among patients treated for laryngeal carcinoma with total laryngectomy. METHOD: Patients with laryngectomy and their family underwent interviews by a sociologist and a head and neck surgeon. RESULTS: The diagnosis of cancer fills the patient's mind with images of suffering and death. Their life trajectory is more often a downward spiral. Patients are so stigmatized by the neoplasic illness that they become marginal and live in social isolation. CONCLUSION: Being laryngectomized completely transforms patients' lives because this surgery induces their topple into the world of the disabled. Total laryngectomy has a negative connotation in our society. Therapists should take this reality into account and intensify their involvement before and especially after surgery. PMID- 19128583 TI - Comparison of closed reduction alone versus primary open repair of acute nasoseptal fractures. AB - OBJECTIVE: Nasoseptal injuries have traditionally been treated via closed reduction. Historically, the high incidence of postreduction deformities has led some surgeons to consider alternative approaches to obtain superior results. Here we compare simple closed reduction versus primary open repair of the nasoseptal fracture. STUDY DESIGN: This was a prospective study of 40 consecutive patients treated with simple closed reduction of their combined nasal bone and septal fracture versus 40 patients treated with closed reduction of their nasal bone fracture and open treatment of the septum. Group outcomes were then compared. RESULTS: In the closed reduction group, 60% had significant postoperative septal deviation, whereas only 12.5% suffered from residual septal deformity in the open group. This resulted in a statistically significant reduction (p < .01) of patients requiring a second operation to formally address the septum. CONCLUSION: By addressing the septum through an open approach, a statistically significant reduction in the number of patients requiring revision rhinoplasty was achieved. PMID- 19128584 TI - Deafness associated with acetaminophen and codeine abuse. AB - Ototoxicity associated with narcotic-acetaminophen combinations is not widely recognized. This can be the cause of severe-to-profound hearing loss that may be overlooked. Otolaryngologists who encounter patients with progressive hearing loss with no apparent cause should specifically ask about overuse of medications containing acetaminophen and a narcotic. Many patients feel that this form of medication is "safe" because it can be purchased over-the-counter. PMID- 19128585 TI - Hyperproliferation in nasal polyposis tissues is not associated with somatic genomic instability. AB - BACKGROUND: Nasal polyps (NPs) are abnormal lesions that arise from any portion of the nasal mucosa or paranasal sinuses within the nasal cavities. They are characterized by the formation of inflammatory swellings of unstable respiratory mucosa, which protrude into the nasal cavity. The causes and the pathogenesis of NP development are not well understood. The current and principal hypothesis is that NPs result from allergy and inflammation, which cause an unstable respiratory mucosa with increased epithelial cell proliferation and epithelial morphologic changes. OBJECTIVE: To understand if epithelial cell hyperproliferation can lead to acquired genomic instability and if nasal polyposis is a consequence of acquired chromosomal alterations in hyperproliferative nasal epithelia. MATERIALS AND METHODS: To test this hypothesis and to perform a survey of chromosomal aberrations in nasal polyps, we applied array comparative genome hybridization (aCGH; 1 Mb resolution, made of 2464 bacterial artificial chromosome (BAC), P1 derived artificial chromosome (PAC), and P1 clones spanning the whole human genome) to deoxyribonucleic acid (DNA) obtained from the NPs of nine patients. The patients recruited in this study had been analyzed for Ki-67 expression using the MIB-1 antibody as a marker of proliferation and were chosen to represent a wide range of hyperproliferative status. RESULTS: After data analysis, no chromosomal aberrations were detected by aCGH. CONCLUSIONS: Our data do not support the hypothesis that nasal polyposis is a consequence of acquired chromosomal alterations in hyperproliferative nasal epithelia. PMID- 19128586 TI - Metastatic renal cell carcinoma to the head and neck: summary of 21 cases. AB - OBJECTIVE: Metastasis of renal cell carcinoma (RCC) to the head and neck is a relatively uncommon phenomenon, but such lesions can be the initial presentation of disease. RCC should be considered a source for lesions in this region, especially in patients who have received a previous diagnosis of RCC. METHODS: In this study, we review the presentation, management, and outcomes for 21 patients with documented metastasis of RCC to the head and neck outside the central nervous system. This study is a retrospective review of cases ascertained during a 17-year period from a large teaching hospital. RESULTS: Of the 21 patients observed, 10 were found to have metastatic disease at the time of RCC diagnosis, involving the head and neck in 5 cases. The most common sites of head and neck metastasis were to bone (n = 6), skin and subcutaneous tissue (n = 6), and lymph nodes (n = 5). CONCLUSIONS: This study is a reminder to consider a renal primary tumour for metastatic disease identified in the head and neck, particularly metastases with a "clear cell" histologic pattern. A head and neck metastasis may occasionally be the presenting sign in a patient with RCC, or it may follow the primary diagnosis by many years. PMID- 19128587 TI - Vestibular evoked myogenic potentials in healthy control subjects using the head rotation method. AB - OBJECTIVE: To evaluate the methodology for vestibular evoked myogenic potential (VEMP) testing, with a focus on the usefulness of the head rotation as a means to tense the sternocleidomastoid (SCM) muscles. Our goal was to obtain normative data for our site and methodology. DESIGN: Prospective study. SETTING: Outpatient otolaryngology clinic. METHODS: P rospective study. Volunteers without an otologic history were selected. While sitting, the volunteer rotated his or her head away from the tested ear so as to tense the ipsilateral SCM. While the SCM was tensed, the volunteer was presented with loud clicks. Responses were averaged. MAIN OUTCOME MEASURES: VEMP positive (pI) wave latency and amplitude, VEMP negative (nI) wave latency and amplitude, and volunteer ability to perform the head rotation method. RESULTS: In healthy controls, average P1 latencies (milliseconds) were 12.58 +/- 0.39 and N1 latencies were 21.88 +/- 0.40. Average P1 amplitudes (microvolts) were 20.48 +/- 1.94 and N1 amplitudes were -25.56 +/- 2.21. VEMP amplitudes were less consistent in patients with limited neck strength. CONCLUSIONS: VEMPs can be elicited from the majority of volunteers. The test is easy to perform, and it is only mildly uncomfortable for most patients using the head rotation method. PMID- 19128588 TI - Extranodal T-cell lymphoma of the sinonasal tract presenting as severe rhinitis: case series. AB - OBJECTIVES: Systemic diseases such as Wegener granulomatosis, Churg-Strauss syndrome, and sarcoidosis can present initially as severe rhinitis alone. One clinical entity that mimics severe rhinitis and thus poses a particular challenge in this regard is extranodal T-cell lymphoma of the sinonasal tract. Sinonasal lymphoma has a vague initial presentation that masquerades as more common, benign causes of rhinitis but progresses as a "midfacial progressive destructive lesion" and is uniformly fatal if untreated. In the absence of systemic involvement, the precise etiology frequently defies diagnosis. We present a series of illustrative cases to raise awareness of this rare yet lethal disease to help reduce the therapeutic delays that are currently encountered clinically. METHODS: A retrospective case series of four patients was reviewed. Each patient presented initially with severe rhinitis but was later diagnosed with extranodal natural killer/T-cell non-Hodgkin lymphoma of the sinonasal tract. RESULTS: The predominant presenting symptoms were unilateral nasal obstruction (four patients), rhinitis (three patients), and facial pain (two patients). Only one patient reported systemic B symptoms. Initial management of all four patients included repeated biopsies revealing nonspecific inflammation, which resulted in extensive therapeutic delays (mean time until diagnosis 21.5 months). The primary outcomes were two deaths and two complete remissions. CONCLUSION: The challenge in making an early diagnosis of T-cell lymphoma of the sinonasal tract has been reaffirmed in our case series. We emphasize that diagnosis requires a high index of clinical suspicion in combination with multiple deep and appropriately processed biopsies that are submitted for immunohistochemistry and molecular studies. PMID- 19128589 TI - Vestibular system in infants after systemic therapy with amikacin. AB - OBJECTIVE: The ototoxic action of systemic therapy with aminoglycosides, used in vital infections in infants, is well recognized. Nevertheless, the routine examination of the infants' inner ear does not include vestibular tests. The purpose of the study was to evaluate vestibular function in a group of infants before and after administration of aminoglycosides, using caloric tests and vestibular evoked myogenic potentials (VEMPs). DESIGN: Randomized clinical study. SETTING: Tertiary referral centre. METHODS: VEMPs and auditory brainstem responses were recorded and caloric stimulation was performed in 68 infants aged 2.5 to 3.5 months: 40 healthy controls and 28 infants after therapy with amikacin. Antibiotic therapy was administered for a respiratory infection in 18 infants and for sepsis in 10 infants. The tests were performed on the day of admission to hospital and on the day of discharge. MAIN OUTCOME MEASURE: The vestibular organ in infants after systemic therapy with amikacin was damaged more frequently than the cochlea. RESULTS: On the day of discharge, no reaction to caloric stimulation was elicited in six patients and no VEMPs were recorded in four subjects. Hearing thresholds were normal in all of the individuals during both examinations. The caloric test was a more sensitive indication of the ototoxic impact of amikacin on the vestibular receptors, followed by VEMPs. CONCLUSIONS: Simultaneous impact of amikacin on the receptor cells of the vestibular organ causes damage to both the semicircular canal and otolith organ receptors. The function of the former is more frequently impaired. The vestibular tests should be performed in infants after treatment with aminoglycosides. PMID- 19128590 TI - Nasal pathophysiology and its relationship to the female ovarian hormones. AB - The influence of the female ovarian hormones on nasal physiology has been the subject of ongoing research, especially over the last 20 years, yet their role remains unclear. This review looks at the evidence to date from the studies that have looked at the different physiologic and therapeutic scenarios in which the ovarian hormones are raised. It also examines the current pathophysiologic links between the hormonal surges and nasal congestion and the missing links that need to be addressed in future studies. PMID- 19128591 TI - The Sunnybrook experience: review of deep vein thrombosis and pulmonary embolism in otolaryngology. AB - OBJECTIVE: To determine the incidence of venous thromboembolism in otolaryngology patients at a Canadian tertiary care hospital. METHODS: This was a retrospective review of all otolaryngology operations at Sunnybrook Health Sciences Centre between January 1, 1989, and December 31, 2004. All patients who developed a deep vein thrombosis (DVT), a pulmonary embolism (PE), or both as in-patient postoperative complications were identified. RESULTS: Fourteen patients were identified in our study. Ten patients (71.4%) developed a DVT, five patients developed a PE (35.7%), and one patient (7.1%) developed both. The overall incidence of DVT and PE was 0.1% and 0.05%, respectively. The highest incidence occurred in patients undergoing major head and neck surgery. Patients who developed a PE had a statistically significantly longer hospital stay than patients who developed a DVT (p < .05). There were two deaths in the five patients who developed a PE. CONCLUSION: Venous thromboembolism is an infrequent but potentially devastating cause of morbidity and mortality in the otolaryngology patient population. PMID- 19128592 TI - Assessment of Toll-like receptor 2 gene polymorphisms in severe chronic rhinosinusitis. AB - BACKGROUND: Chronic rhinosinusitis (CRS) is believed to reflect an inflammatory response of the sinonasal mucosa to bacteria and/or fungi. Staphylococcus aureus, a gram-positive organism, is frequently implicated. Toll-like receptor 2 (TLR2) is involved in innate immunity, recognizing gram-positive organisms via detection of bacterial lipopeptides. As a poor response to sinus surgery has been associated with reduced levels of TLR2 expression, and given the frequent recovery of S. aureus in this condition, we suspected that polymorphisms in TLR2 genes are implicated in this condition. OBJECTIVE: To investigate the association between single nucleotide polymorphisms (SNPs) in the TLR2 gene and CRS. METHODS: Two hundred six patients with severe CRS and 200 controls were recruited prospectively. A maximally informative set of SNPs in the gene encoding TLR2 were selected from the HapMap data set and genotyped. RESULTS: Eleven of 12 SNPs were successfully genotyped. No significant associations could be detected for the SNPs tested within the limitations of our study, which has the power to detect only those SNPs with a relative risk of 2.0 or greater. CONCLUSIONS: Our findings do not support a role for polymorphisms in the TLR2 gene in the pathogenesis of CRS. Nevertheless, other genetic variants within genes regulating innate immunity may be involved and will require further assessment. PMID- 19128593 TI - Effect of increasing duration of stimulation on the electrically evoked auditory brainstem and middle latency responses in pediatric cochlear implant users. AB - OBJECTIVE: Electrophysiologic responses used to predict behavioural stimulation levels in implant users are typically evoked with a single-pulse stimulus versus higher-rate pulse trains of longer duration. Unfortunately, electrophysiologic measures tend to overestimate behavioural responses. It may be possible to improve the predictive ability of evoked potential thresholds by increasing the duration of the stimulus. We asked if auditory brainstem and middle latency responses can be reliably measured using electrical pulse trains of increasing duration. DESIGN: Evoked potential testing was performed in five pediatric N24RE cochlear implant users aged 7.6 to 14.9 years. SETTING: Cochlear Implant Program, Cochlear Implant Laboratory, The Hospital for Sick Children, Toronto. METHODS: Responses were electrically evoked for durations of 2, 6, and 10 milliseconds by single pulses and pulse trains of 500 pulses per second. MAIN OUTCOME MEASURES: Evoked auditory brainstem response (EABR) and evoked middle latency response (EMLR) waveform latencies and amplitudes were compared between durations. RESULTS: Clear and replicable EABRs and EMLRs were recorded for all durations. There was no significant change in EABR wave eV latency relative to the stimulus offset. There was no significant change in EMLR wave latencies relative to the stimulus onset. This was confirmed by a linear decrease in the interwave latency between the EABR wave eV and the EMLR wave eNa. No significant changes in wave amplitude were found as the pulse train duration increased for the EABR or the EMLR. CONCLUSIONS: EABRs are dominated by the stimulus offset, whereas EMLRs appear to be dominated by stimulus onset. Clear, unchanging EABRs and EMLRs indicated that electrophysiologic measures were a valid tool for increasing durations. PMID- 19128594 TI - Linking laryngopharyngeal reflux to otitis media with effusion: pepsinogen study of adenoid tissue and middle ear fluid. AB - HYPOTHESIS/OBJECTIVES: Laryngopharyngeal reflux (LPR) was recently implicated in the etiology of otitis media with effusion (OME). To study the hypothesis that gastric juice reaches the middle ear through the nasopharynx and eustachian tube, we evaluated the presence of pepsinogen in the adenoid tissues of children with otitis media with effusion (OME) and compared them with the tissues of a control group of children without OME. METHODS: In the study group, middle ear effusions (MEEs) and adenoidal tissue biopsies were obtained from patients undergoing simultaneous tympanostomy tube placement and adenoidectomy. In the control group, adenoid specimens were taken during adenoidectomy (+/- tonsillectomy) from children with no history of OME. The adenoid tissues were analyzed immunohistochemically to confirm the presence of pepsinogen. Normal gastric tissue was used as a positive control and lymphatic tissue as a negative control. Total pepsinogen levels of MEE were measured with enzyme-linked immunosorbent assay. RESULTS: Adenoid tissue of the OME group (n = 25) demonstrated significantly higher pepsinogen immunoreactivity when compared with the adenoid tissue of the control group (n = 29), specifically in staining of both the epithelia (p < .0001) and the subepithelia, (p < .001). The presence of pepsinogen was detected in 84% of MEEs from the OME group, at concentrations 1.86 to 12.5 times higher than that of serum. Pepsinogen messenger ribonucleic acid was not demonstrated in any of the adenoid tissues of the OME group. CONCLUSION: LPR plays an important role in the pathogenesis of OME as gastric reflux reaches the middle ear through the nasopharynx and eustachian tube to cause OME. PMID- 19128595 TI - Treatment results of carcinoma in situ of the glottic larynx: 61 patients treated with radiotherapy. AB - OBJECTIVES: To review the Notre-Dame Hospital experience in the treatment of carcinoma in situ of the glottis by radiotherapy and to evaluate the different factors affecting local control and survival. METHODS: Between January 1990 and June 2002, 61 patients presenting with carcinoma in situ of the glottis were treated with curative intent radiotherapy. No patients received either surgery or chemotherapy in the initial treatment of their cancer. RESULTS: The median follow up for the entire population was 3.2 years. The local control rate was 96% and 94% at 2 and 5 years, respectively. Overall survival was 100% and 90% at 2 and 5 years, respectively. No statistically significant prognostic factor could be identified either for local control or survival. No patient experienced severe treatment complications or death. CONCLUSION: Radiotherapy offers excellent treatment results for carcinoma in situ of the glottic larynx, with few treatment complications. These results are comparable to those published in the literature and justify our choice of primary radiotherapy for carcinoma in situ of the glottis. PMID- 19128596 TI - Comparative study of posttonsillectomy hemorrhage with the use of diclofenac versus dihydrocodeine for postoperative analgesia and review of the literature. AB - OBJECTIVE: To assess the effect of postoperative diclofenac on the posttonsillectomy hemorrhage rate in adults. DESIGN: Retrospective chart review with Caldicott Guardian approval of patients before and after a change in the postoperative drug regimen. One hundred ninety-three adults between 16 and 56 years, American Society of Anesthesiologists Grade I, listed for elective tonsillectomy, were included in the analysis. SETTING: A tertiary referral centre in the United Kingdom. METHODS: One hundred ten adults had cold dissection tonsillectomy and had been prescribed soluble diclofenac postoperatively. This group was compared with 83 adult patients who underwent cold dissection tonsillectomy and had been prescribed dihydrocodeine solution postoperatively. Our hypothesis was that the postoperative use of diclofenac made no difference to the incidence of secondary hemorrhage. The Fisher exact test was used for analysis. MAIN OUTCOME MEASURES: Comparison of a previously reported posttonsillectomy hemorrhage rate with the rate for the year after the addition of diclofenac. The occurrence of primary or secondary postoperative hemorrhage and its management was recorded. RESULTS: Three of 110 (2.73%) patients who used postoperative diclofenac had secondary hemorrhage compared with 6 of 83 (7.23%) of those patients treated with dihydrocodeine. The Fisher exact test for secondary hemorrhage therefore gives a p value of .131, and there is no significant difference. CONCLUSIONS: Diclofenac is a useful and safe addition to postoperative analgesia for adult tonsillectomy. There was an apparent reduction in secondary hemorrhage with the postoperative use of diclofenac. PMID- 19128597 TI - Endoscopic treatment of Cis-T2 glottic cancer with a CO2 laser: 2-year survival analysis of 36 cases. AB - OBJECTIVE: To report our 2-year survival outcomes for early glottic carcinoma using microsurgical resection with the CO2 laser. DESIGN: A prospective study. METHODS: Outcomes from a cohort of 36 patients with glottic cancer staged Cis-T2 (7 Cis, 17 T1, 12 T2) who underwent endoscopic resection using a CO2 laser between January 2002 and June 2005 were studied. The mean follow-up was 27 months (range 0-54 months). All follow-ups were performed at our multidisciplinary head and neck oncology clinic at Dalhousie University, Halifax, Nova Scotia. RESULTS: Using a Kaplan-Meier survival analysis, the 2-year recurrence-free survival for the cohort was 89%, with a standard error of 6.7%. At the time of the study, no patient had died of laryngeal cancer, one patient had died of other disease, and one patient was lost to follow-up. There had been three recurrences in the cohort, two local recurrences (T1), and one recurrence in a regional lymph node (T2). After salvage, all patients with recurrences were disease free at the time of the study. Of 35 cases with follow-up, 60% had no reported problems with voice on their last visit and 17% reported consistent hoarseness or weakness. CONCLUSION: These results justify the continued treatment of early glottic carcinoma with CO2 laser microsurgery at our centre. PMID- 19128598 TI - Velopharyngeal insufficiency clinic: the first 18 months. AB - OBJECTIVES: To profile the presentation and management of patients seen at a velopharyngeal insufficiency (VPI) clinic and to explore the role of commonly used investigations in the workup of VPI. DESIGN: Retrospective review. SETTING: Tertiary care VPI clinic. METHODS: A computerized database created at the time of patient assessment was accessed to review 75 patients seen over an 18-month span. MAIN OUTCOME MEASURES: Descriptive analysis of patient demographics, presenting complaints, pathophysiology, and treatment. The incidence of chromosome 22q microdeletion in patients presenting to such a clinic was also determined. RESULTS: The most common etiology was persistent VPI following prior cleft palate surgery. Eleven percent of presenting patients were identified with 22q microdeletion by fluorescent in situ hybridization testing. CONCLUSIONS: VPI is the result of a wide number of etiologies, with a high incidence of 22q microdeletion identified. Routine genetic testing in VPI clinics is advocated. PMID- 19128599 TI - Autoimmune inner ear disease: clinical and laboratory findings and treatment outcome. AB - OBJECTIVE: To assess the demographics, clinical presentations, auditory and vestibular findings, value of serologic tests, and treatment outcome in autoimmune inner ear disease (AIED). STUDY DESIGN: Retrospective chart review. SETTINGS: Tertiary care centre. PATIENTS: Sixty patients with confirmed AIED, with and without systemic disease. INTERVENTIONS: Diagnostic auditory, vestibular, and serologic tests and treatment with steroids. OUTCOME MEASURES: Auditory, vestibular, and serologic findings and treatment outcomes. RESULTS: The female to male ratio was 2:1. Forty-nine patients presented with unilateral or bilateral hearing loss; 28 patients also had vestibular symptoms. Eleven patients had vestibular symptoms only. Hearing loss was progressive in most, rapid in onset in one, and of sudden onset in two. Approximately 25% of patients had confirmed systemic autoimmune disease. Patients without systemic disease had serologic tests to confirm the diagnosis of AIED. The level of antinuclear antibodies was high, with a speckled pattern, in 38 patients, and 9 patients had a high rheumatoid factor. The positive yield of other detailed tests was low. Vestibular tests showed a peripheral type of change. Steroid treatment produced an excellent response in 33% and a good response in 16% without systemic disease. Only 25% of those with systemic disease had a similar response. Patients with vestibular symptoms only had an excellent response to steroids. CONCLUSIONS: Patients with AIED present with varied symptoms, and some have only vestibular symptoms. Limited serologic tests used in the diagnosis of systemic autoimmune diseases are valuable in establishing the diagnosis of AIED. Fifty percent of patients with AIED have an excellent response to steroids. Those with systemic disease have a lower response rate. Those with vestibular symptoms only are responsive to steroids. PMID- 19128600 TI - Socioeconomic factors in head and neck cancer. AB - OBJECTIVE: To evaluate the impact of socioeconomic status on the incidence of head and neck cancer using a population-based survey. METHODS: We employed pooled individual-level data from the US National Institutes of Health Survey for the years 1997 to 2006 inclusive. We performed a logistic regression analysis for four variables of socioeconomic status (marital status, family income, highest level of education achieved, immigration status) and four potential confounding variables (age, race, smoking status, alcohol consumption). The effects of these socioeconomic variables on head and neck cancer were compared with their effects on cancers overall. RESULTS: There was a statistically significant increase in head and neck cancer incidence for adult men with the following characteristics: status as single, never married and education less than high school completion. There was a trend toward higher rates of head and neck cancer with annual family income < $20,000 US. No such associations were seen for cancer in general. CONCLUSIONS: There is evidence to support the contention that individuals with more disadvantaged socioeconomic status have higher rates of developing head and neck cancer, even after controlling for associated health behaviours such as smoking and alcohol consumption. This work suggests that further study into the effects of socioeconomic deprivation and head and neck cancer is warranted. PMID- 19128601 TI - Is stuttering triggered by the insertion of tympanostomy tubes? PMID- 19128602 TI - Surgical management of the labyrinthine fistula complicating chronic otitis media with or without cholesteatoma. AB - OBJECTIVES: To present our most recent experience on the diagnosis and treatment of the labyrinthine fistula. The relative data are compared with those of our past experiences adopting the same modality of presenting the results. METHODS: The clinical and surgical records of 334 patients affected with chronic otitis media with or without cholesteatoma were analyzed to evaluate the frequency, site, and size of the labyrinthine fistulae. In this study, a more aggressive strategy was adopted that comprised immediate total removal of the cholesteatoma matrix and the surrounding inflammatory tissues even when they involved the membranous labyrinth. RESULTS: Sixteen labyrinthine fistulae were found, only one of which was in a patient with chronic otitis media without cholesteatoma. Only one patient suffered from postoperative hearing deterioration of the bone conduction above 10 dB. The fistulae were smaller in size than in the previous survey, although the total incidence was slightly higher. CONCLUSIONS: Imaging techniques have demonstrated a favourable clinical impact on the diagnosis regarding the size and involvement of one or more anatomic structures of the otic capsule, not on the frequency percentage of fistulae. Total removal of the cholesteatoma matrix in one step, also combined with partial labyrinthectomy, yields satisfactory hearing results. To obtain successful outcomes, it is essential to respect certain fundamental precautions. PMID- 19128603 TI - Impact of pulsatile nasal irrigation on the prognosis of functional endoscopic sinus surgery. AB - OBJECTIVES: Nasal irrigation is a popular adjunctive therapy after sinonasal surgery. The objective of this study was to determine the efficacy of nasal irrigation after functional endoscopic sinus surgery (FESS). METHODS: Patients with chronic rhinosinusitis (CRS) who underwent FESS were enrolled. The preoperative computed tomography (CT) Lund-Mackay staging, Lund symptom, and endoscopy scores were assessed. All patients received postoperative endoscopic debridement and medical treatment and were randomly allocated to receive nasal irrigation for 3 months or not. The symptom and endoscopy scores were repeatedly measured at 2 weeks and 1, 2, and 3 months postoperatively. RESULTS: Seventy seven subjects completed the study. When patients were classified as having either severe or mild CRS (CT score > 12 or < or = 12, respectively), the patients with mild CRS in the irrigation group had a significantly better endoscopy score and symptom score change rates than those in the nonirrigation group (p = .019 and .032, respectively), but there were no significant differences in those outcomes between irrigation and nonirrigation groups in patients with severe CRS. CONCLUSIONS: Our results showed that nasal irrigation is an effective post-FESS management in patients with mild CRS. However, the efficacy of nasal irrigation in patients with severe CRS needs further investigation. PMID- 19128604 TI - Aesthetic reconstruction of a crooked nose via extracorporeal septoplasty. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness, indications, and contraindications of the extracorporeal approach in treating a severely deviated nose. METHODS: Sixty-four patients operated on for aesthetic correction of a severely deviated nose were studied. Forty-six of them were first operations and 18 were revision cases. The surgical procedure of choice was the closed technique. Septal cartilage was prepared and then dislocated, followed by external alignment and reimplantation. Hump reduction with lateral osteotomies and, occasionally, medial dislocation of nasal bones was carried out. RESULTS: Cosmetic results were satisfactory in all first operation cases. In 11 of the 18 revision cases, septal preparation was impossible; in 2, septal sagging was observed; and in 1, perforation was noted. CONCLUSIONS: Extracorporeal septoplasty is an effective, safe, and reliable technique, especially for twisted noses undergoing surgery for the first time. However, it is strongly contraindicated in a revised deviated nose. PMID- 19128605 TI - The microdebrider and its complications in endoscopic surgery for nasal polyposis. AB - OBJECTIVE: The aim of this study was to assess the rate of complications of the microdebrider in endoscopic sinus surgery (ESS) for nasal polyposis. STUDY DESIGN AND SETTING: A retrospective study was done on the complications occurring during ESS for nasal polyposis and chronic rhinosinusitis (CRS) without nasal polyps in which a microdebrider had been used on 275 sides of 139 patients in a tertiary university practice setting. RESULTS: The mean sinus opacification scores in computed tomography, according to the Lund-Mackay staging system, were 9.2 +/- 2.28 for the nasal polyposis group and 4.8 +/- 1.54 for the CRS group. There was one major complication, a cerebrospinal fistula that was repaired during the surgery. The minor and major complication rates for the nasal polyposis group were 11.8% and 0.5%, respectively. The minor complication rate for the CRS group was 4%. CONCLUSION: Knowledge of endonasal anatomy, surgical experience, a bloodless operating field, and being careful about every colour change during the operation are the fundamental essentials for lowering the complication rate. With its lower incidence of complications, even in high-risk cases such as nasal polyposis, a microdebrider provides acceptable complication rates in ESS. Also, the severity of the disease is the main determining factor for the final complication rate. PMID- 19128606 TI - Clinical predictors of peritonsillar abscess in adults. AB - OBJECTIVE: The primary objective was to identify clinical predictors that differentiate a peritonsillar abscess from peritonsillar cellulitis in adults. The secondary objective was to identify the prevalence of tobacco smoking behaviour in this group of patients with peritonsillitis. SETTING: The Ottawa Hospital, a tertiary care centre. METHODS: Retrospective chart review of patients diagnosed with peritonsillar abscess or peritonsillar cellulitis in the adult emergency department of The Ottawa Hospital during the years 2002 to 2004. RESULTS: The charts of 130 patients with peritonsillitis were reviewed. Patients' ages ranged from 16 to 91 years. Fifty patients were diagnosed with a peritonsillar abscess and 80 with peritonsillar cellulitis. Forty-four percent of the patients in this study admitted to active tobacco smoking behaviour. Clinical signs significantly associated with peritonsillar abscess included trismus (p < .001), uvular deviation (p < .001), and inferior displacement of the superior pole of the tonsil (p < .001) on the affected side. Pain duration was not a significant discriminative factor (p = .069). CONCLUSIONS: The diagnosis of a peritonsillar abscess is based on clinical findings. In this study, the prevalence of tobacco smoking behaviour in patients with peritonsillar disease was high. Further study is recommended to investigate the potential causal effects of tobacco smoke on the development of peritonsillar disease. PMID- 19128607 TI - Massive lymphatic malformations of the head, neck, and chest. AB - OBJECTIVE: To provide clinicians with an overview of the complications and functional and clinical outcomes of children affected by massive lymphatic malformations affecting multiple sites in the head, neck, and/or chest. METHODS: We reviewed the medical records of all patients who received treatment for massive cervicofacial lymphatic malformations at our tertiary care pediatric institution during a 17-year period. Massive lesions were defined as (1) unilateral lesions that infiltrated at least 50% of two anatomic regions, including the head, neck, or chest; (2) unilateral lesions in which there was involvement of the midline structures of the head or neck; or (3) bilateral lesions in which 50% of each side was affected. Data consisted of critical clinical parameters, including extent of anatomic involvement, treatment interventions, pre- and post-treatment complications, lesion resolution, and functional outcomes. RESULTS: Sixteen patients were treated for massive head, neck, and chest lymphatic malformations. Ten of 16 had massive unilateral lesions, whereas 6 had bilateral lesions. Eleven patients experienced airway compromise resulting from airway involvement; eight of these patients required placement of a tracheostomy tube. Six patients, all of whom had airway compromise, suffered with orodigestive complications; four of these six required gastrostomy tube placement. Eleven of 16 patients were managed exclusively with surgery, and 3 of 11 showed marked (60-80%) disease resolution; however, all 3 of these patients required preoperative placement of a tracheostomy tube, and only 1 was decannulated following resection. The severity of the airway symptoms resulted in the death of three patients, all of whom had tracheostomies. CONCLUSIONS: Children with massive head, neck, and chest lymphatic malformations suffer from high morbidity and significant mortality. Airway involvement is an important prognostic factor in these patients. PMID- 19128608 TI - Use of autologous dermal graft in the treatment of parotid surgery wounds for prevention of neck scars: preliminary results. AB - BACKGROUND: Reconstruction after parotid surgery still represents a challenge for the otolaryngologic surgeon, particularly to avoid neck scars and Frey syndrome. The interposition of fascia lata grafts and sternocleidomastoid muscle flaps represents the most common surgical solutions that have been suggested, even if these options often increase local morbidity. To date, the most exciting and promising progress in reconstruction of postparotidectomy wounds comes from the application of bioengineered tissues. METHODS AND OBJECTIVE: This was a prospective preliminary study. The results of treatment of parotid surgery wounds with the use of the autologous dermal graft TISSUEtech autograft system (Fidia Advanced Biopolymers, FAB, SrL, Abano Terme, Padua, Italy) are presented. RESULTS AND CONCLUSIONS: The postoperative follow-up results from this study demonstrate efficacy in preventing depression or neck scars after parotid surgery using the TISSUEtech autograft system. PMID- 19128609 TI - Effectiveness of royal jelly on tympanic membrane perforations: an experimental study. AB - OBJECTIVE: The aim of this study was to evaluate the comparison of the efficacy of royal jelly in a guinea pig tympanic membrane perforation model with untreated and control groups. STUDY DESIGN: Prospective study using 16 adult guinea pigs. METHODS: Each animal underwent subtotal removal of the tympanic membrane in one ear. Animals were divided into two groups of eight animals each. Group A was treated with topical royal jelly, and group B was treated with saline. All opposite right ears were used as the control group. Examination was done by otoendoscopy on days 10, 12, and 14. The healing process was observed under the microscope and photographed for 3 months. RESULTS: There was no significant difference between group A and group B in the incidence of perforation closure. However, after 3 months, the mean thickness of the tympanic membranes was 0.11, 0.07, and 0.08 mm, respectively, in group A, group B, and the control. These results showed a statistically significant difference. Histologically, the tympanic membranes of group A showed marked fibroblastic orientation and well organized connective tissue. Application of royal jelly improved the success rate of closure, as shown in the present study. CONCLUSIONS: This is the first study to investigate the effectiveness of royal jelly in enhancing healing in tympanic membrane perforations. Our study suggests that royal jelly is effective in increasing healing of tympanic membrane perforations in guinea pigs. PMID- 19128610 TI - Comparison of swabs versus suction traps for endoscopically guided sinus cultures. AB - BACKGROUND: Knowledge of the causative organism(s) in bacterial rhinosinusitis has become the cornerstone of adequate medical and surgical management. Little uniformity and data exist for the best method of obtaining sinus cultures. Most otolaryngologists tend to use the nasal swab for obtaining transnasal middle meatal cultures. A prospective study was carried out to compare the effectiveness of standard nasal swabs versus suction traps in obtaining bacterial isolates under endoscopic guidance. METHODS: Fifty-two patients with purulence in the middle meatus or frontal recess were included in the study. All patients were cultured using nasal wire swabs. Twenty-five of these patients also had the purulence suctioned into a Xomed Sinus Secretion Collector (XSSC) (Xomed Surgical Products, Jacksonville, FL), and 27 patients had the purulence suctioned into a standard Leukens trap (Busse Hospital Disposables, Hauppauge, NY). All specimens were sent to the hospital microbiology laboratory within 1 hour of capture. RESULTS: The average number of bacteria cultured per patient was 1.21 for the swab, 1.37 for the XSSC trap, and 1.08 for the Leukens trap. The agreement between bacteria captured from the wire swab and suction trap was 76.9%, with significant agreement being observed by all isolates with the exception of coagulase-negative Staphylococcus and the other gram-positive bacteria group consisting of Streptococcus viridans, beta-hemolytic streptococcus, and alpha hemolytic streptococcus. CONCLUSION: This study suggests that the wire swab appears to be as efficacious at obtaining endoscopically guided cultures as the Xomed and Leukens suction devices. PMID- 19128611 TI - Detection of Helicobacter pylori in nasal polyps: preliminary report. AB - OBJECTIVE: Investigation of Helicobacter pylori (HP) colonization in the nasal polyp specimens of patients with chronic rhinosinusitis (CRS). DESIGN: A prospective clinical trial. SETTING: The study was performed in 23 adult patients with sinonasal polyposis and 15 controls with concha bullosa (CB) who underwent endoscopic sinus surgery. METHODS: In all patients, gastroesophageal reflux (GER) symptoms were evaluated, and in 30 of them, the HP status of the stomach was also studied. Nasal polyps, mucosa of the middle concha, and gastric specimens were analyzed using a urease test and by Giemsa staining. MAIN OUTCOME MEASURES: We found HP in nasal polyp specimens in six CRS patients by the urease test and by histochemical analysis with Giemsa staining. RESULTS: All of them had HP in gastric specimens and GER symptoms. HP was not detected in any patient with CB. CONCLUSIONS: These results may have implications for a possible role of HP in CRS with polyposis. If a patient has HP in his gastric mucosa and is positive for GER symptoms, HP may be found in polyps as well. Whether HP is one of the causative agents of CRS or is a result of CRS is not known. Further studies are needed to find an answer to these questions. PMID- 19128612 TI - Survey of current perspectives on laryngopharyngeal reflux among Canadian otolaryngologists. AB - PURPOSE: Laryngopharyngeal reflux (LPR) -- gastroesophageal reflux above the upper esophageal sphincter -- is a common problem encountered by otolaryngologists. Despite consensus guidelines, the presentation, diagnosis, and treatment remain controversial. We surveyed Canadian otolaryngologists to assess current perspectives. METHODS: Web-based questionnaires were e-mailed to 135 otolaryngologists. Respondents were categorized by subspecialty as head and neck (H&N) or non-H&N (rhinology, otology, laryngology, facial plastics, general and pediatric otolaryngology). Data were analyzed to determine differences in proportions between groups. RESULTS: The response rate was 48 of 135 otolaryngologists. Symptoms considered to be strongly or moderately associated with LPR included globus sensation, excessive throat clearing, sore or burning throat, hoarseness, chronic cough, and dysphonia. The laryngoscopic signs considered strongly associated with LPR were edema, intra-arytenoid changes, and granulomata. The majority of otolaryngologists in both the H&N (12 of 15) and non H&N groups (27 of 32) use flexible laryngoscopy for investigation and diagnosis of LPR. Proton pump inhibitors in addition to lifestyle modifications are recommended by both groups as first- and second-line therapy for an initial course of 6 to 12 weeks, with long-term therapy extended for 4 to 12 months. CONCLUSION: Canadian otolaryngologists do correlate specific signs and symptoms with LPR patients. This is consistent across subspecialties within the field. Flexible fibre-optic laryngoscopy is the preferred diagnostic tool. Although evidence based on randomized controlled trials has yet to demonstrate a reproducible, statistically significant improvement in LPR from medical therapy, first-line pharmacologic treatment (in addition to lifestyle changes) is generally provided as a proton pump inhibitor, with the duration of therapy being somewhat variable and less than that recommended by the current literature. PMID- 19128613 TI - Home-based humidification for mucositis in patients undergoing radical radiotherapy: preliminary report. AB - INTRODUCTION: Oropharyngeal mucositis is a frequent, severe complication of local irradiation for tumours in the head and neck. We postulated that heated humidification of inspired air via a nasal interface may palliate symptoms of mucositis by reducing the discomfort associated with dry, sticky secretions. We sought to review the effect of home-based humidification on hospital admissions and the patient reported experience of that humidification. METHODS: This study was a retrospective review. A historical (control) group of patients did not receive home humidification at any stage (n = 55) and a study group (n = 53) received home humidification at or after the onset of grade 3 mucositis. A questionnaire was sent to study group patients to obtain information about their experience of using the humidifier at home. RESULTS: There were no demographic differences between the study and control groups, but the study group had significantly more advanced cancer (stage IV; p = .0307) and significantly higher total fractions and days treated (p < .01). Group comparison showed no difference in subsequent overall hospital admissions (p = .9269), but 7 of the 55 control group patients (12.7%) were admitted for supportive care within 2 months of completing radiotherapy, whereas none of the 53 patients who used home humidification were admitted after starting that use (p < .01). Almost all (95%) of the study group patients reported that humidification was of benefit, and 81% stated that it relieved mouth or throat pain. CONCLUSION: Humidification of inspired gas offers a simple, drug-free option for managing a number of the adverse mucosal effects of radiation and chemoradiation in head and neck cancer patients. PMID- 19128614 TI - Use of tissue sealant for day surgery parotidectomy. AB - OBJECTIVES: To evaluate the use of tissue sealant in facilitating day surgery parotidectomy without the use of surgical drains and to consider the potential economic benefit using this technique. STUDY DESIGN AND SETTING: Prospective cohort study of 21 patients undergoing parotidectomy for nonmalignant disease in a university hospital. Surgery as a day procedure without the use of surgical drains was planned. The costs associated with parotidectomy, including the use of tissue sealant and its delivery system, versus in-patient admission with a drain were calculated and compared. METHODS AND OUTCOME MEASURES: Parotidectomy was undertaken by one surgeon. Prior to wound closure, the skin flap and wound bed were approximated using Tisseel tissue sealant (Baxter Corp., Mississauga, ON). Data regarding the costs of the tissue sealant, the delivery system, and hospital in-patient stay were obtained to enable an economic comparison. Patients were followed to assess surgical outcome and document any complications. RESULTS: There were no major surgical complications. One patient required admission for control of postoperative nausea. None of the patients felt that discharge had been premature. The estimated cost advantage of this technique applied to institutions in Canada was $1,775 per case. CONCLUSIONS: Parotidectomy can be undertaken safely in a day surgery setting without the need for surgical drains. The increased cost associated with the use of tissue sealant compared with surgical drains is greatly overshadowed by the economic advantage of undertaking day surgery. There is a significant potential cost saving to the health care system. PMID- 19128615 TI - Management of inner nasal valve insufficiency. AB - OBJECTIVE: To determine the efficacy of the upper lateral caudal edge management in treating patients with inner nasal valve insufficiency and collapse. DESIGN: This was a prospective study in a group of patients suffering from nasal obstruction owing to nasal valve insufficiency. Forty-three patients, who had undergone septoplasty and inferior turbinectomy, were included. All patients presented with nasal obstruction resulting in persistent functional problems. An anatomically narrow nasal valve, valve collapse, or both were found during the clinical examination. SETTING: A tertiary referral centre. METHODS: Revision was undertaken using a new technique to restore the nasal valve, based on upper lateral caudal edge management. MAIN OUTCOME MEASURES: Improvement in nasal airway patency evidenced by patient questioning, by clinical inspection of the nose, and by rhinomanometric results. RESULTS: None of the patients had major complications. In 28 (65.1%) patients, symmetrically improved nasal airway patency and elimination of the subjective sensation of inspiratory collapse was found. In 12 (27.9%) patients, a remarkable asymmetric improvement in nasal obstruction was evident. Two (4.6%) patients reported a moderate breathing improvement, and in only one (2.3%) patient, revision was needed. The mean follow up time was 18 months. CONCLUSIONS: Our proposed method is an effective therapeutic approach in the management of inner nasal valve insufficiency. It reconstitutes the normal tension of the inner nasal valve and reestablishes the stiffness and resistance of the lateral nasal wall. It can be performed under local anesthesia in cooperative patients, with minimal morbidity and a high rate of success. PMID- 19128617 TI - Risk of aspiration following radiation for non-nasopharyngeal head and neck cancer. AB - OBJECTIVE: We assessed the rate of aspiration following radiation for non nasopharyngeal head and neck cancer. DESIGN: Retrospective study. SETTING: Veterans Administration Hospital. PATIENTS: Thirty-three patients who underwent radiation for head and neck cancer. Modified barium swallow was performed prior to and following treatment to assess the persistence of dysphagia and aspiration risk. All patients were cancer free at the time of the swallowing study. Dysphagia severity was graded from 1 to 7. RESULTS: Preradiation baseline dysphagia was observed as follows: 10 grade 1, 14 grade 2, 9 grade 3, and 1 grade 4. Following radiation, at a median follow-up of 3 months, nine patients had grade 1, eight patients had grade 2, six patients had grade 3, two patients had grade 4, three patients had grade 5, two patients had grade 6, and three patients had grade 7. Overall, 24% (8 of 33) of the patients developed aspiration (grades 5-7). Fifteen percent (5 of 33) of the patients had severe aspiration (grades 6 7) requiring tube feedings. All patients who developed severe aspiration continued to require tube feedings more than 1 year following treatment completion. CONCLUSION: Aspiration is a significant source of morbidity following radiation for non-nasopharyngeal head and neck cancer. Aspiration may develop for all tumour stages or sites. Diagnostic studies such as modified barium swallow should be included in future prospective head and neck cancer studies to assess the prevalence of aspiration because of its often silent nature. PMID- 19128616 TI - Prevalence of pharyngeal and esophageal stenosis following radiation for head and neck cancer. AB - OBJECTIVE: To evaluate the risk and outcome of pharyngoesophageal stenosis in patients who complained of dysphagia following radiation for head and neck cancer. DESIGN: Retrospective study. SETTING: Veterans Administration hospital. PATIENTS: Patients who complained of persistent dysphagia following radiation alone or combined with surgery or chemotherapy for head and neck cancer. Patients were selected if they were cancer free at the time of the swallowing study. All patients had modified barium swallow (MBS) and an endoscopic examination for initial evaluation of their dysphagia. Traditional barium swallow was requested when there was a suspicion of pharyngoesophageal stenosis on MBS. RESULTS: Two hundred twenty-two patients underwent MBS for evaluation of dysphagia posttreatment. Traditional barium swallow confirmed the diagnosis of pharyngeal (n = 2) or esophageal (n = 14) stenosis in 16 patients. Eight patients had esophageal stenosis on endoscopic examination. All patients underwent dilatation for relief of their dysphagia. The number of dilatations performed was, respectively, one in 12 patients, two in 4 patients, three in 3 patients, four in 3 patients, five in one patient, and six in one patient. CONCLUSION: Pharyngeal and/or cervical esophageal stenosis may be the cause of dysphagia following radiation for head and neck cancer. Esophageal dilatations often offer temporary relief of the dysphagia. PMID- 19128618 TI - Vastus lateralis muscle-only free flap: defining its role in head and neck reconstruction. AB - OBJECTIVE: To review the anatomy, indications, technique, advantages, and limitations of vastus lateralis muscle-only free tissue transfer for reconstruction of head and neck defects following tumour ablation. DESIGN: A single-centre retrospective chart review was undertaken. METHODS: Patients undergoing free tissue transfer of the vastus lateralis muscle for reconstruction of head and neck defects between May 1, 2003, and April 30, 2006, were identified. Demographic data, surgical details, outcome, and complications were determined. RESULTS: Nine patients were identified. A variety of tumour types and sites were treated. Three suffered flap-related complications consisting of distal flap vascular compromise. CONCLUSIONS: The vastus lateralis muscle-only free flap has numerous advantages, including versatility, ease of harvest, and ability to perform a two-team operation. There are also some limitations, including variable anatomy and tenuous distal circulation. Judicious use of this free flap in noncritical areas is preferable. PMID- 19128619 TI - Obstructive upper airway problems and primary enuresis nocturna relationship in pediatric patients: reciprocal study. AB - OBJECTIVE: To determine the relationship between primary enuresis nocturna (PEN) and obstructive upper airway problems (OUAPs). DESIGN: Prospective, descriptive study. SETTING: Tertiary care academic medical centre. METHODS: Two groups of patients were included in the study. Newly diagnosed pediatric PEN patients between August 2004 and June 2005 were evaluated for OUAPs in the Department of Otorhinolaryngology of our institution and constituted the first group of patients. During the same period, patients who were on the surgery list for upper airway problems were examined for PEN in the Department of Pediatric Psychiatry as the second group. Obtained data were analyzed with regard to patient demographics and the relationship between PEN and OUAPs, and follow-up results are reported. MAIN OUTCOME MEASURES: There were 32 patients in each group. In the first group, 5 patients (15.6%) had totally obstructing severe septal deviation, 8 patients (25.0%) had moderate septal deviation, 10 patients (31.3%) had allergic rhinitis and inferior turbinate hypertrophy, 3 patients had adenotonsillar hypertrophy (9.4%), and 1 patient (3.1%) had septal perforation. The frequency of OUAP in PEN patients was 65.6%. In the second group, 10 patients (31.3%) had a PEN diagnosis. After at least 3 months (mean 6.3 months) of follow up, four patients (40%) were free of PEN and two patients (20%) experienced improvement in PEN. Another four patients did not have any change in the severity of PEN. CONCLUSIONS: According to our results, OUAPs are very common in PEN patients and PEN is a common symptom of OUAPs. PMID- 19128620 TI - Systematic review and meta-analysis assessing the effectiveness of local anesthetic, vasoconstrictive, and lubricating agents in flexible fibre-optic nasolaryngoscopy. AB - BACKGROUND: Flexible fibre-optic nasolaryngoscopy (FFN) is a common otolaryngology procedure that patients may find uncomfortable. Preparative agents, including topical anesthetic, vasoconstrictive, and lubricating agents, have been studied in randomized controlled trials (RCTs). METHODS: A systematic review and meta-analysis was conducted on RCTs published between January 1966 and October 2005 and indexed to MEDLINE, CINAHL, and Cochrane CENTRAL databases. Methodologic validity was evaluated. Primary outcomes were patients' evaluation of FFN. Secondary outcomes were endoscopists' evaluation of FFN. RESULTS: Eight RCTs were identified studying five preparative agent classes: vasoconstrictors plus topical anesthetics, vasoconstrictors alone, topical anesthetics alone, lubricating agents, saline, and no treatment. The systematic review found no difference in patients' evaluation of pain and discomfort for cocaine versus Co phenylcaine (two RCTs), Co-phenylcaine versus no active agent (three RCTs), topical anesthetics versus no active preparative agent (two RCTs), vasoconstrictors versus no active preparative agent (two RCTs), and lubricating agents versus nothing. Two RCTs found that Co-phenylcaine causes higher taste unpleasantness, and one RCT found that topical anesthetics cause higher pain. Lubricating agents increase ease of examination but decrease quality of view (one RCT). Meta-analysis of two studies comparing Co-phenylcaine with other preparative agents found no difference on pain scores. CONCLUSIONS: Cumulative evidence from eight RCTs shows no difference in pain scores when preparative agents with vasocontrictive, topical anesthetic, or lubricating properties are used. Co-phenylcaine may cause higher taste unpleasantness, and lidocaine may cause more pain. For the endoscopist, lubricating agents may aid in the examination but compromise the quality of the view. PMID- 19128621 TI - Ethical considerations in resource allocation in a cochlear implant program. AB - OBJECTIVE: To review processes of resource allocation and the ethical considerations relevant to the fair allocation of a limited number of cochlear implants to increasing numbers of potential recipients. DESIGN: Review of relevant considerations. SETTING: Tertiary referral hospital. METHODS: Editorial discussion of the ethical issues of resource allocation. MAIN OUTCOME MEASURES: Heterogeneity of audiometric thresholds, self-reported disability of hearing loss, age of the potential cochlear implant recipient, cost-effectiveness, access to resources, compliance with follow-up, social support available to the recipient, social consequences of hearing impairment, and other recipient-related factors. RESULTS: In a publicly funded health care system, there will always be a need for decision-making processes for allocation of finite fiscal resources. All candidates for cochlear implantation deserve fair consideration. However, they are a heterogeneous group in terms of needs and expected outcomes consisting of traditional and marginal candidates, with a wide range of benefit from acoustic amplification. CONCLUSIONS: We argue that implant programs should thoughtfully prioritize treatment on the basis of need and potential benefit. We reject queuing on the basis of "first-come, first-served" or on the basis of perceived social worth. PMID- 19128622 TI - Uvulopalatopharyngoplasty and uvulopalatal flap for the treatment of snoring: technique to avoid complications. AB - OBJECTIVE: To review the outcome of 50 snoring patients treated with uvulopalatopharyngoplasty and uvulopalatal flap, looking for a decrease in the surgical complications rate. DESIGN: A longitudinal, prospective, self-controlled pilot study. SETTING: Sleep Disorders Clinic of the National University of Mexico and Angeles Lomas Hospital. MATERIAL AND METHODS: Fifty consecutive patients underwent surgical treatment for snoring. The Muller manoeuvre was performed in all of them to determine the site of obstruction. In all cases, the bed partner was asked about the presence of snoring before and 6 months after the procedure. Polysomnography was performed before and after the surgery to assess sleep apnea in all patients and to determine the success regarding the snoring index. Every complication was registered. RESULTS: All patients had only habitual snoring, without sleep apnea. Thirty-eight males and 12 females were included in the study. At the sixth postoperative month, the bed partner of 48 patients (96%) reported that snoring was gone, 2 (4%) said that snoring was no longer a problem, the initial snoring index mean was 214, and 6 months after the procedure it decreased to 12. No patients had any complication related to the surgical procedure. CONCLUSIONS: Despite the fact that there are a number of techniques for snoring and sleep apnea, we achieved good postoperative results while avoiding the most common complications. PMID- 19128623 TI - Complications of tongue base somnoplasty. AB - BACKGROUND: There are many therapeutic modalities to treat obstructive sleep apnea, and somnoplasty is one of the most widely used around the world. However, there are few reports about its complications. METHODS: The charts of consecutive patients who underwent tongue base somnoplasty were reviewed in the search for complications. RESULTS: The total number of patients was 237. We found only 2 postoperative complicated cases. DISCUSSION: Tongue base somnoplasty can be considered a safe procedure with a low incidence of complications. PMID- 19128624 TI - Results of intratympanic dexamethasone injection as salvage treatment in idiopathic sudden hearing loss. AB - OBJECTIVE: To evaluate the efficacy of intratympanic (IT) dexamethasone injection as a salvage treatment in idiopathic sudden hearing loss (ISHL) patients. DESIGN: A prospective study. SETTING: A secondary care hospital. METHODS: Fifty-two subjects whose pure-tone averages (PTAs) after a 2-week standard treatment were > 30 dB or worse than 10 dB from the contralateral ear were included. Thirty-four patients underwent IT dexamethasone injection and 18 patients (control) received no further treatment. Final hearing outcome was compared between the two groups. MAIN OUTCOME MEASURES: At 2 weeks after IT dexamethasone injection, audiograms were obtained and compared between the two groups. RESULTS: Sixteen patients (47.1%) in the IT dexamethasone group and 8 patients (44.4%) in the control group finally showed an improvement of 10 dB or more in PTA or a 15% or more change in speech discrimination score (p > .05). The average improvement in PTAs was 8.2 dB in the IT dexamethasone group and 3.6 dB in the control group (p = .184). Patients responsive to the initial treatment showed better final outcomes, irrespective of IT dexamethasone injection. CONCLUSIONS: The present study does not support IT dexamethasone injection as a beneficial salvage treatment for ISHL patients. The true efficacy of this procedure still needs to be investigated. PMID- 19128625 TI - Surgical treatment of rhinophyma. AB - RATIONALE: Rhinophyma is a slowly progressive, disfiguring disease of the nose. A variety of surgical techniques have been described for its management, which have had varying success. OBJECTIVE: To describe the technique and results of the senior author (P.B.) using electrocautery decortication and/or CO(2) laser contouring in seven consecutive patients with rhinophyma by retrospective review. DESIGN: A retrospective case series of seven consecutive adult patients treated from 1999 to 2005 inclusive. METHODS: A retrospective chart review was conducted of the seven patients treated at the local tertiary care centre over a 6-year period. RESULTS: In the seven consecutive patients who were operated on by the senior author, all had been previously diagnosed with acne rosacea. Five patients had moderate and two had major rhinophyma. Five patients (71%) had an excellent or very good result using the described technique. CONCLUSION: Electrocautery nasal decortication and/or CO(2) laser contouring in patients with rhinophyma is an effective technique that results in a satisfied patient with an aesthetically suitable nose. Caution should be exercised when vaporizing tissue in the nasal alar region with a CO(2) laser to prevent postoperative alar retraction. PMID- 19128626 TI - Informed consent in otologic surgery: prospective randomized study comparing risk recall with an illustrated handout and a nonillustrated handout. AB - OBJECTIVE: Following consent, do pictures in a handout improve patients' recall of otologic surgical risks? STUDY DESIGN: Prospective, randomized trial in a tertiary care centre. METHODS: Patients undergoing otologic surgery were consented with a standardized checklist of risks by two surgeons. They were randomized (stratified by educational level) to receive either a pure text or a text and pictures handout outlining the risks of surgery. A telephone interview tested recall at a mean of 19 days. Twenty-six patients were resampled at 1 year. MAIN OUTCOME MEASURES: Recall was analyzed with respect to type of handout, age, sex, and level of education. A subset of 31 patients was analyzed for the effects of which surgeon consented, previous otologic surgery, and actually reading the handout. RESULTS: Fifty-one patients completed the study. The overall risk recall was 43%, with 45% in the pictorial group and 42% in the pure text group (p = .84). The illustrated handout did not improve the recall of any individual surgical risk either. Higher education improved risk recall from 36 to 54% (p = .009). Age, consenting surgeon, previous otologic surgery, and even reading the handout did not improve risk recall. A subgroup of 26 patients was followed up 1 year later, and their recall fell from 41 to 35%. The illustrated handout did not improve long-term recall (p = .674). CONCLUSION: Pictorial cues do not improve recall of surgical risks, but education level does. PMID- 19128627 TI - Pain management after tonsillectomy: morphine is not enough. AB - INTRODUCTION: Tonsillectomy is usually done as day surgery, transferring to the parents most of the postoperative care. They often feel alone and incompetent in front of a child in pain. We developed a guide to help parents for both the evaluation and the management of postoperative pain. OBJECTIVE: To improve pediatric posttonsillectomy pain management with the implementation of a parental guide and treatment algorithm. METHODS: Eighty children (5-17 years) were recruited in this randomized clinical trial. Parents and children were assigned to one of the research groups: control or experimental. All received an identical prescription for acetaminophen (15 mg/kg) and morphine (0.3 mg/kg), information about the medication, a visual analogue scale (VAS), and index cards to be filled at home. The experimental group also received a treatment algorithm associating pain scores with analgesics. RESULTS: Pain scores for the control group and the experimental group were 5.3 and 4.7 on the day of surgery, 4.0 and 3.6 the next day, 2.2 and 2.3 3 days later, and 1.6 and 1.4 1 week after the intervention, respectively. The side effects and the analgesics given were comparable for both groups. CONCLUSION: No significant difference was demonstrated for the studied parameters, probably because the majority of the parents followed the prescriptions well. The guide was much appreciated. The children understood the VAS easily and liked using it. The combination of acetaminophen-morphine needs to be revisited since pain remains problematic. PMID- 19128628 TI - Evaluation of nasal obstruction with Odiosoft-Rhino in nasal septal deviation. AB - BACKGROUND: Evaluation of a recently developed method for measuring nasal obstruction. OBJECTIVE: To investigate the reliability of a new software program called Odiosoft-Rhino (OR) as a presumptive diagnostic method to evaluate the nasal airflow of patients who have nasal septal deviation and to compare OR results with the acoustic rhinometry (AR) results and visual analogue scale (VAS) of nasal obstruction. SETTING: Referral centre, institutional practice. METHODS: OR uses a software program to analyze the original sounds of nasal airflow and evaluate the sounds of both nasal inspiration and expiration and then performs spectral and frequency analysis. OR and AR were performed on 56 patients with nasal septal deviations and 52 healthy controls. Their VAS, minimal cross sectional area (MCA)1, and MCA2 measurements and nasal expiratory sound analysis between 200 to 500, 500 to 1000, 1000 to 2000, 2000 to 4000, and 4000 to 6000 Hz frequencies were noted for the left and right nasal cavities. MAIN OUTCOME MEASURE: Assessment and comparison of nasal obstruction with OR and AR methods. RESULTS: There was a significant difference between the MCA1 and MCA2 values and the nasal expiratory sound intensity of the deviated patients at 200 to 500, 500 to 1000, 1000 to 2000, and 2000 to 4000 Hz of the patient group and the control group (p < .001). There was a correlation between the OR results of 2000 to 4000 and 4000 to 6000 Hz intervals and the MCA1 results of the deviated sides. From the receiver operating characteristic curve analysis, the pooled results showed that 81.8% of deviated patients in the 2000 to 4000 Hz frequency range were found under the curve (p < .001). Related to this, 83.4% sensitivity and 82.1% specificity with a 14.5 dB cutoff point were determined. CONCLUSION: The OR test is noninvasive and requires little cooperation. The sensitivity and specificity of the OR test are reliable, so we could propose to use OR as a new diagnostic method to evaluate the nasal airflow in clinical practice. However, more studies with wider series and some technical modification are needed. PMID- 19128629 TI - Value of lateral neck radiography for ingested foreign bodies using the likelihood ratio. AB - OBJECTIVE: To determine the diagnostic accuracy of lateral neck radiography and its value in related radiographic findings in the detection of foreign bodies using the likelihood ratio. PATIENTS AND METHODS: This retrospective study comprised 114 patients who had neck radiography followed by rigid esophagoscopy. Interpretations of the radiographs were correlated with esophagoscopic findings and clinical follow-up for at least 1 month. Data were mainly assessed by likelihood ratio and detection rate. RESULTS: Plain radiography had a positive likelihood ratio of 2.4 (95% confidence interval [CI] 1.8-3.1) and a negative likelihood ratio of 0.6 (95% CI 0.40-0.7). It had a sensitivity of 57.4% (95% CI 45.6-69.1) and a specificity of 76.1% (95% CI 63.8-88.4). Positive likelihood ratios for each radiographic finding (foreign body shadow, air, and soft tissue swelling) were 3.4 (95% CI 2.6-4.4), 1.5 (95% CI 0.9-2.6), and 4.1 (95% CI 1.9 8.7), respectively. Plain radiography had a high detection rate (72.5%) when foreign bodies were lodged in the hypopharynx and cervical esophagus. On the contrary, the detection rate was poor (0%) in the oropharynx. CONCLUSIONS: A lateral neck radiograph is better taken after a negative finding in a thorough oral examination and fibre-optic laryngoscopy. Judging from the possible site of impacted foreign body and radiographic findings, we can better predict the presence of foreign bodies. PMID- 19128631 TI - Analysis of the efficacy of marketing tools in facial plastic surgery. AB - OBJECTIVES: To compare referral sources to a facial plastic surgery practice and to develop models correlating the referral source with the decision for surgery. DESIGN: Retrospective descriptive study. SETTING: Well-established, metropolitan, private facial plastic surgery practice with training fellowship affiliated with an academic centre. METHODS: One-thousand eighty-nine new consecutive patients presenting between January 2001 and December 2005 recorded intake data including age, gender, and chief complaint. Final data input was their decision for or against surgery. MAIN OUTCOME MEASURES: Main outcome measures included differences in referral sources based on data collected and how those sources related to decision for surgery. RESULTS: A 50% conversion rate was found. Women and older patients were more likely to be referred from magazines, television, and newspapers and for facial rejuvenation. Men and younger patients were more likely to be referred from the website and for rhinoplasty. For facial rejuvenation, both the number of patients interested in and the probability that they agreed to the procedure increased with age. For rhinoplasty, the converse was true. The most likely patients to schedule surgery were those who were referred from other patients, friends, or family members in our practice. CONCLUSIONS: The data confirm that word-of-mouth referrals are the most important source for predicting which patients will elect to proceed with surgery in this established facial cosmetic surgery practice. PMID- 19128632 TI - Predictors of unexpected hospital admissions after outpatient endoscopic sinus surgery: retrospective review. AB - BACKGROUND: Endoscopic sinus surgery (ESS) is typically performed on an outpatient basis in our centre. The purpose of this study is to determine the frequency of unexpected stays in patients who undergo ESS. This information would be useful to identify and to counsel these potential patients. METHODS: Retrospective chart review of 194 consecutive patients who had ESS during a 6 month span. Multivariate analysis was performed on 11 variables identified in the charts to determine whether any of the data predicted an unexpected admission. RESULTS: The unexpected rate of admission was 4.7%. Surgical complications causing admission occurred in 1% of the cases. Reasons for admissions included nausea and vomiting, hypotension, oxygen desaturation, headache, postoperative epistaxis, and observation for possible cerebrospinal fluid leak. None of the 11 variables that were examined showed statistical significance as independent predictors of unexpected admissions. However, two variables, the presence of comorbidities and the use of ondansetron, did approach statistical significance. CONCLUSIONS: Unexpected admissions following ESS are infrequent, and the reasons for admission are varied. In this study, although 9 of the 11 parameters evaluated showed no statistical significance as independent predictors of unexpected admissions, 2 did approach statistical significance. Intuitively, the presence of patient comorbidities would be expected to place the patient at greater risk of unexpected admission. Ondansetron is a potent antiemetic and is reserved for patients at risk or in those having severe symptoms. These patients would also be expected to have an increased risk of an unanticipated hospital stay. PMID- 19128633 TI - Pharyngocutaneous fistula after total laryngectomy: emphasis on lymph node metastases as a new predisposing factor. AB - OBJECTIVE: We investigated patient-related and local risk factors for pharyngocutaneous fistula developing after total laryngectomy. DESIGN: Retrospective clinical study. SETTING: Tertiary medical centre. METHOD: The study included 255 patients who underwent total laryngectomy and concurrent neck dissection between July 2001 and February 2006 for laryngeal cancer. MAIN OUTCOME MEASURES: The association of fistula and patient-related (age, sex, hemoglobin and albumin levels, diabetes mellitus, chronic obstructive pulmonary disease, chronic congestive heart disease, hypertension) and local risk factors (T stage of the tumour and localization, previous radiotherapy, preoperative tracheostomy, lymph node metastases, surgical margin) was analyzed using the chi-square and independent-sample t-tests. RESULTS: Pharyngocutaneous fistulae were found in 18.4% of the patients. Lymph node metastases, chronic congestive heart disease, and postoperative hemoglobin and albumin levels lower than 10.84 +/- 1.2 g/dL and 3.06 +/- 0.5 g/dL, respectively, were found to be statistically significant risk factors in fistula formation. The logistic regression model was carried out on these risk factors. CONCLUSION: Pharyngocutaneous fistula remains the most frequent complication of total laryngectomy despite improved preoperative assessment and postoperative care. Lymph node metastases emerged for the first time as a risk factor for fistula formation. Our experience confirmed that meticulous postoperative care should be paid to the patients who have contributing risk factors and lymph node metastases. PMID- 19128634 TI - Evaluation of the influence of visual stimulation in the active mechanisms of the organ of Corti by analyzing the changes in the amplitude of distortion products. AB - INTRODUCTION: The central nervous system can regulate the input of information to the cochlea by means of the activity of the olivocochlear efferent system on the active micromechanisms of the organ of Corti. This article discusses a quantitative study of the inhibitory phenomenon that visual tasks exert on active cochlear micromechanisms. MATERIAL AND METHOD: We studied prospectively the characteristics of distortion products (DPs) in 80 normal ears, with and without visual stimulation. RESULTS: The results showed a decrease in the amplitude of DPs (2F1-F2), which was significant in frequencies between 1500 and 6000 Hz. CONCLUSION: Visual stimulation causes a modulation of the cochlear micromechanisms from the central nervous system from the medial olivocochlear bundle of the efferent auditory pathway. PMID- 19128635 TI - Blood loss in endoscopic sinus surgery: assessment of variables. AB - OBJECTIVE: To identify the variables that influence blood loss in endoscopic sinus surgery performed under uniform conditions of preoperative topical vasoconstriction and surgical technique. STUDY DESIGN: Blind, nonrandomized, observational study. SETTING: One hundred thirty-eight consecutive patients who underwent endoscopic sinus surgery with the same technique in a university hospital over a 9-month period. METHODS: Pearson product-moment correlation was used for numerical variables and nonparametric tests for categorical variables: Mann-Whitney (comparison of two independent variables) and Kruskal-Wallis (comparison of more than two independent variables). MAIN OUTCOME MEASURES: Operative time, total blood loss, and blood loss per minute were correlated with blood loss markers: age, sex, surgeon, type and severity of illness, septoplasty, and anesthetic agents used. RESULTS: No correlation was found between age, sex, surgeon, and blood loss markers. Severe polyposis produced more blood loss than mild polyposis and sinusitis. Septoplasty showed an interesting inverse correlation with blood loss. Remifentanyl with fluorinated volatile agents was accompanied by less blood loss than other anesthetic agents. CONCLUSION: More blood loss occurred with more severe sinonasal pathology. Blood loss may be reduced by using remifentanyl with a fluorinated volatile agent and, possibly, preoperative infiltration with epinephrine. PMID- 19128636 TI - Isolated vertigo and dizziness of vascular origin. AB - INTRODUCTION: Vertigo of vascular origin is usually limited to migraine, transient ischemic attacks, and ischemic or hemorrhagic stroke. Excluding migraines, focal neurologic deficits usually allow a fast identification of the impending neural damage and permit a rapid transfer to a stroke team. Few authors have considered that a vascular origin should be thought of in cases of positional vertigo or vertigo not associated with neurologic signs (ie, isolated vertigo) in patients with vascular disease risk factors. OBJECTIVES: (1) To present a case series of vertigo and dizziness without vertigo of probable vascular origin and (2) to perform an evidence-based review regarding the incidence of isolated vertigo and dizziness of vascular origin. DESIGN: Retrospective nonrandomized case series. SETTING: Tertiary referral centre. METHOD: (1) All patients from a vertigo clinic found to have vertigo of potential vascular origin had a complete neuro-otologic examination and were investigated by magnetic resonance angiography. (2) A MEDLINE literature search was performed using specific search terms to identify pertinent publications concerning this pathology. They were reviewed and graded according to the quality of their evidence. RESULTS: We identified nine cases of vertigo and dizziness without vertigo of probable vascular origin. We identified 27 818 potentially pertinent articles. A detailed review yielded 13 studies for further analysis. Although isolated vertigo has frequently been reported as the only symptom in case series of cerebellar infarcts, only one article discussed its incidence. CONCLUSION: Vascular origin should be considered in cases of positional vertigo and isolated vertigo or dizziness when the etiology remains unclear. A single study has determined that 52% of patients with isolated vertigo of unclear etiopathology have posterior circulation anomalies. PMID- 19128637 TI - Retrospective review of 242 consecutive patients treated surgically for parotid gland tumours. AB - OBJECTIVE: To compare the trends and specific outcomes of patients treated surgically for parotid neoplasms between 1994 and 2005 with those of patients who were treated between 1985 and 1993 at Mount Sinai Hospital in Toronto and to identify preoperative indicators of malignancy. METHODS: A retrospective chart review was performed that included all patients who received treatment from 1994 to 2005 at Mount Sinai Hospital. The results of this review was compared with those of another group at the same institution. RESULTS: The study included 242 patients. Of these patients, 183 (75.6%) had benign neoplasms, 51 (21.1%) had malignant neoplasms, and 8 (3.3%) had inflammatory or lymphatic lesions. The most common associated symptoms were swelling (17.4%) and pain (16.5%). Only 5.8% of patients presented with facial weakness; however, the likelihood of malignancy was high (64.3%). Total parotidectomy was reserved for 26.4% of patients; the majority (68.2%) received a subtotal parotidectomy. Common postoperative complications included temporary facial nerve palsy (23.6%), Frey syndrome (12.4%), and salivary fistula (5.0%). Analysis of the correlation between fine needle aspiration and final cytology revealed that fine-needle aspiration sensitivity and specificity were 60.0% and 96.1%, respectively. Comparison of these results with the other study at our institution revealed no significant differences in patient predictors or outcomes. CONCLUSIONS: These results were compared with those of a previous study conducted at the same institution between 1985 and 1993. There was no significant deviation from former trends. The presenting symptoms of facial nerve weakness and lymphadenopathy were found to be strong indicators of malignant disease. Facial pain was found not to be a reliable prognostic feature for malignancy. PMID- 19128638 TI - Preoperative localization in primary hyperparathyroidism. AB - OBJECTIVE: To determine the most effective preoperative localization techniques for patients with primary hyperparathyroidism to facilitate the surgical procedure, decrease patient morbidity, and decrease the number of repeat surgeries owing to inability to locate the abnormal parathyroid gland. METHODS: This was a retrospective study in which 53 patients with primary hyperparathyroidism underwent preoperative sestamibi scanning and ultrasonography. If the two tests failed to agree on the precise location of the abnormal gland, a third imaging technique, magnetic resonance imaging (MRI), was used to confirm the precise location of the gland. Patients with secondary, tertiary, and recurrent hyperparathyroidism and patients with thyroid carcinoma were excluded from this study. Twenty males and 33 females were involved in the study. The mean age was 59.8 years (range 34-84 years). The preoperative results were compared with findings in surgery. A successful surgery was defined as parathyroid hormone and corrected calcium values in the normal range following the operative procedure. RESULTS: There was concurrence between ultrasound and sestamibi scanning in 70% (37 of 53) of the patients. When both agreed, the identified location of the abnormal parathyroid gland was correct 97% (36 of 37) of the time. The ultrasound and sestamibi scanning did not coincide in 30% of the patients (16 of 53). In this scenario, MRI was performed. When the MRI agreed with either of the two previous imaging techniques, the abnormal gland was accurately localized 100% of the time. In six cases (11%), there was no definitive agreement between all three tests that were performed. CONCLUSION: The combination of preoperative ultrasonography and sestamibi scanning is effective in predicting the location of parathyroid adenomas in patients with primary hyperparathyroidism. When both tests conflict, MRI is an effective tool to localize the abnormal glands. This study describes an algorithm for the preoperative localization of parathyroid gland abnormalities, in particular parathyroid adenomas. Second, it allows patients to undergo unilateral neck exploration, as opposed to bilateral neck exploration, where operative times, duration of hospitalization, and patient morbidity are potentially decreased. PMID- 19128639 TI - Adjustable laryngeal implant for unilateral vocal cord paralysis. AB - BACKGROUND: Laryngeal framework surgery has been widely accepted as a definitive treatment for unilateral vocal cord paralysis (UVCP). An adjustable laryngeal implant has been developed for precise medialization of the paralyzed vocal cord. OBJECTIVES: To describe the implant and surgical technique and to evaluate the effect of vocal cord medialization using an adjustable laryngeal implant on the quality of life of patients with UVCP. PATIENTS AND METHODS: Fifty-three patients with UVCP who had undergone medialization with the adjustable laryngeal implant were identified. All patients completed the Voice Handicap Index (VHI) quality of life questionnaire. Preoperative and postoperative scores were compared. RESULTS: Major advantages over other accepted implants are the stability of the implant, excellent biocompatibility, precise medialization with a micrometric screw, and ease of secondary adjustment. Mean VHI score in all handicap domains was significantly improved following vocal fold medialization (p < .01). Although no difference was found in regard to gender, younger patients had higher handicap scores than older patients preoperatively (p < .05). This difference was not present following medialization. CONCLUSIONS: The adjustable laryngeal implant offers many advantages over other techniques. Vocal cord medialization using the adjustable laryngeal implant significantly improves quality of life as measured by the VHI. PMID- 19128640 TI - Early recognition of nasopharyngeal cancer in adults with only otitis media with effusion. AB - OBJECTIVE: To assess the probability of nasopharyngeal carcinoma (NPC) being present among adults who appeared to suffer only from otitis media with effusion (OME). DESIGN AND SETTING: The clinical features of 87 adults suffering from OME but who demonstrated no other apparent symptoms and signs suggestive of NPC at the Department of Otorhinolaryngology, Kaohsiung Medical University Hospital, between January 2003 and December 2004, were analyzed. A routine nasopharyngeal biopsy was performed. RESULTS: Five patients revealed NPCs (four nonkeratinizing carcinomas and one squamous cell carcinoma). OME was deemed to have resulted from upper respiratory infection in 20 patients (23.0%), chronic paranasal sinusitis in 15 patients (17.3%), allergic rhinitis in 16 patients (18.4%), NPC in 5 patients (5.7%), various other known etiologies in 11 patients (12.6%), and unknown etiologies in 20 patients (23.0%). The incidence of NPC among adults suffering from OME but featuring no other symptoms and signs suggestive of NPC was 5.7% (5 of 87 patients). Such a figure appeared to be greater than that in some other reports because NPC tends to feature a greater prevalence in Taiwan. Therefore, biopsy of the postnasal space for adults who appear to feature only OME when NPC is strongly suspected is necessary for the early diagnosis of NPC. CONCLUSIONS: We conclude that Taiwanese adults with only OME, the cause of which is unclear, should be subjected to an appropriate medical examination and biopsy of the nasopharynx to exclude NPC. PMID- 19128641 TI - Epigenetic perspective into head and neck cancer through in silico gene expression profiling of histone lysine methyltransferases. AB - OBJECTIVE: To investigate the role of histone lysine methylation in normal and cancerous head and neck tissues, which will provide novel insight into the pathophysiology of oncogenesis in these tissues. DESIGN: Gene expression profiles from histone lysine methyltransferases (HKMTases) were obtained from UniGene expressed sequence tags (ESTs). HKMTase expression levels in respective tissues were analyzed for elevated expression or altered levels of normal versus human cancers. SETTING: UniGene EST data were derived from normal and cancerous human tissues submitted to UniGene. METHODS: The expression levels of 47 HKMTases were obtained for 46 normal and 25 cancerous tissues, which included 10 normal and 5 cancerous head and neck tissues. An analysis of expression levels was carried out to identify HKMTases with tissue-specific expression and alteration in head and neck cancer. MAIN OUTCOME MEASURES: Expression levels of HKMTases in normal and cancerous tissues of the head and neck in comparison with other anatomic sites. RESULTS: In 7 of 10 head and neck tissues, we identified HKMTases that were specifically elevated in these tissues in comparison with all 46 normal tissues. In all five head and neck cancers, we identified HKMTases specifically elevated in these tumours. In addition, we identitified HKMTases with altered expression in tumour versus normal head and neck tissues. CONCLUSIONS: Normal head and neck tissues have unique epigenetic profiles demarcated by distinct HKMTase gene expression. We identified HKMTases as potentially tissue-specific epigenetic regulators of the head and neck, which could behave as oncogenes or tumour suppressor genes important in malignant transformation of these tissues. PMID- 19128642 TI - Efficacy of a home-based exercise program on benign paroxysmal positional vertigo compared with betahistine. AB - OBJECTIVE: To determine the efficacy of a home-based exercise program by comparing it with betahistine in patients with benign paroxysmal positional vertigo (BPPV). STUDY DESIGN: Prospective, randomized, controlled study. SETTING: Outpatient clinic of a university hospital. PATIENTS: Thirty-eight patients (10 males, 28 females; mean age 46 +/- 13 years) diagnosed as having BPPV. INTERVENTIONS: Patients were randomly assigned to either an exercise or a medication group. In the medication group, betahistine was prescribed at 24 mg/d for 1 month. The exercise group was required to perform Cawthorne-Cooksey exercises six times/day for 4 weeks. MAIN OUTCOME MEASURES: The Vertigo, Dizziness, Imbalance Questionnaire (VDI), which consists of two subscales, the VDI symptom subscale (VDI-ss) and the VDI health-related quality of life (VDI HRQoL) subscale, and the Vertigo Symptom Scale (VSS) were used for assessment at the beginning of the study and after 2 months. RESULTS: The mean scores of the two components of the VDI and the VSS decreased in the exercise group by the fourth week. In the medication group, VSI mean scores, VDIss mean scores, and VSS mean scores decreased in the second week and VDI-HRQoL mean scores decreased in the fourth week. However, there were no significant differences between baseline and week 8. There were significant differences between groups regarding the change in the mean scores of the VDI (p = .001) and the VSS (p = .001) at the end of the study in favour of the exercise group. CONCLUSIONS: Exercise was found to be a better treatment choice than medication and may be preferable for patients with persistent or chronic vertigo. PMID- 19128643 TI - The DizzyFix: initial results of a new dynamic visual device for the home treatment of benign paroxysmal positional vertigo. AB - OBJECTIVE: To develop and test a completely new dynamic visual device for the home treatment of benign paroxysmal positional vertigo (BPPV). DESIGN: Randomized, controlled, prospective trial. SETTING: Tertiary care hospital. METHODS: We designed and manufactured a new device (the DizzyFix) to assist in the performance of the particle repositioning manoeuvre (PRM). Fifty healthy volunteers were taught the PRM, half using the new device. At 1 week, we compared the PRM performance between the device and nondevice user groups. MAIN OUTCOME MEASURES: Performance of the PRM as graded on an 11-point scale. RESULTS: DizzyFix users in phase I scored significantly higher on their PRM performance compared with controls (p = .0001). CONCLUSIONS: The use of this new device enables volunteers to conduct a correct PRM on their own. This is a significant improvement from written instructions or in-office training. PMID- 19128644 TI - Prevalence of new-onset vestibular symptoms following cochlear implantation. AB - OBJECTIVE: To identify the prevalence of new-onset balance symptoms in adult patients who have undergone cochlear implantation (CI). DESIGN: A retrospective, questionnaire-based prevalence study of 227 consecutive adults (age > 16 years) who have undergone CI. The control group comprised patients awaiting CI. SETTING: Tertiary hospital, Vancouver, British Columbia. METHODS: All patients implanted at the study centre were contacted. A questionnaire regarding their past medical history and the presence and character of pre- and post-CI dizziness was completed. Patients were divided into four groups based on the presence or absence of their symptoms in relation to the time of implantation. Patients with vestibular symptoms further completed the Dizziness Handicap Inventory (DHI). MAIN OUTCOME MEASURES: Incidence of balance symptoms pre- and post-CI and associated DHI scores. RESULTS: One hundred ten of 227 patients (48%) responded. Fifty-three (48.3%) had dizziness prior to CI and 64 patients (58.2%) postimplantation. Forty-one patients (37.3%) noted new onset of balance symptoms or a change in their symptoms post-CI. The onset was immediate (within 1 week post-CI) in the majority of this group (63.4%). DHI scores were generally low across all groups. CONCLUSIONS: Delayed- and late-onset new balance-related symptoms occur in approximately 5 and 9% of CI patients, respectively, and the self-perceived handicapping effects of these symptoms may be greater compared with patients with symptoms prior to implantation. PMID- 19128645 TI - Systemic absorption of topical gentamicin sinus irrigation. AB - OBJECTIVE: Evidence surrounding systemic absorption of gentamicin during intraoperative irrigation of the paranasal sinuses is lacking. The objectives of this study were to determine (1) if topical gentamicin is absorbed from the paranasal sinuses, (2) if hearing loss occurs following topical administration of gentamicin, and (3) if gentamicin placed within the sinuses travels retrograde (against mucociliary clearance) up the auditory tube to the middle ear. DESIGN: Consecutive, prospective case series. SETTING: Tertiary centre. METHODS: A series of patients undergoing sinus surgery were identified. Fluorescein-stained gentamicin was used to irrigate the sinus cavities intraoperatively. Otoscopy using a filtered light source was performed 30 minutes postoperatively. MAIN OUTCOME MEASURES: (1) Serum gentamicin levels preirrigation and at 30 minutes postirrigation and (2) change in pure-tone average and threshold at 8 kHz pre- and postoperative audiograms. RESULTS: Twenty patients were enrolled. Serum gentamicin levels were detectable in three patients. Fluorescein irrigation solution was not visualized in the middle ear space. No significant hearing loss was observed in any of the patients. CONCLUSIONS: Gentamicin may be absorbed from the nasal mucosa during intraoperative irrigation of the sinuses. However, detectable serum levels were well below therapeutic levels. The clinical significance of this finding requires further study to determine if topical sinus irrigation with gentamicin is a safe procedure. PMID- 19128646 TI - Vestibular schwannoma: how much residual hearing is useful? AB - OBJECTIVE: The objective of this study was to determine the usefulness of residual hearing in the tumour ear in vestibular schwannoma patients. DESIGN: A prospective case series study. SETTING: The study was performed at the Queen Elizabeth Health Sciences Centre in Halifax, Nova Scotia. METHODS: Thirty-three vestibular schwannoma patients and 13 controls underwent QuickSIN (Etymotic Research Inc, Elk Grove, IL) speech-in-noise testing with the tumour and good ears open and occluded. Nine testing conditions used three speakers with speech signal from the tumour side, the front, and the good ear side, with noise in other speakers. MAIN OUTCOME MEASURES: Tumour ear contribution (TEC) was calculated by the decrease in score with the index ear occluded. Multiple regression analysis and correlation coefficients were used to determine predictors of TEC. RESULTS: The strongest correlation was between the pure-tone average (PTA) and the TEC with signal from the tumour side. The speech discrimination score (SDS) was also significantly correlated with TEC in this condition. Neither PTA nor SDS correlated well with TEC with signal from other directions. Multiple regression analysis with TEC and sound from the tumour ear as the dependent variable showed that the SDS and PTA of the tumour ear are significant independent predictors (p = .049 and .037, respectively). There are no obvious breakpoints in the PTA or SDS to make 50%, 50 dB, or other operating points "special." CONCLUSION: The main contribution of residual hearing is in signals from the tumour side. The various rules are more or less equivalent in discriminating between those who will have a high TEC and those who will not. PMID- 19128647 TI - Donor-site morbidity following radial forearm free tissue transfer in head and neck surgery. AB - OBJECTIVE: To comprehensively evaluate long-term quantitative and qualitative donor-site morbidity following radial forearm free tissue harvest. DESIGN: A single-centre retrospective cohort study with internal controls was undertaken. METHODS: Quantitative measurements of range of motion (ROM) of the forearm, wrist, and digits were performed. Grip and pinch strength and hand dexterity were also evaluated. Qualitative assessment was performed using the Michigan Hand Outcomes Questionnaire (MHQ), a validated quality of life instrument. MAIN OUTCOME MEASURES: Quantitative primary outcome measures were (1) wrist flexion and extension, (2 forearm pronation and supination, and (3) hand dexterity. The qualitative primary outcome measure was overall MHQ score. RESULTS: The operated side demonstrated decreased hand dexterity (p = .008), with no change in wrist and forearm ROM. An increase in ROM of the little finger was found (p = .002). The MHQ demonstrated a perceived decrease in function (p = .031), an increase in pain (p = .045), and no difference in appearance (p = .486). CONCLUSIONS: The radial forearm free flap results in measurable quantitative changes in hand function and limited changes in patient perception. Donor-site appearance does not seem to be an important factor. PMID- 19128648 TI - Patients' quality of life, reported difficulties, and benefits following surgery for acoustic neuroma. AB - OBJECTIVES: We compared the quality of life of patients with acoustic neuroma (AN) with general population controls and other chronic illness patients. We also examined the types and significance of ongoing difficulties reported by AN patients and the types of positive benefits following their surgery. DESIGN: A comparative questionnaire survey. METHODS: A research questionnaire was sent to all 119 members of the Acoustic Neuroma Association of New Zealand. Responses were compared with those of matched general population controls and type 2 diabetes, recent angina, and dermatitis patients. MAIN OUTCOME MEASURES: The questionnaire contained the SF-36, a measure of the type of difficulties following surgery, as well as a measure of illness benefits. RESULTS: AN patients reported an impaired quality of life compared with general population controls and other chronic illness groups, but AN patients reported better physical and general health outcomes. Hearing was the most frequent reported difficulty following surgery, and this impacted most on social functioning. The majority of patients named at least one positive outcome from their illness. AN patients who had facial difficulties following surgery were less likely to report positive outcomes. CONCLUSIONS: Treatment for AN results in a significant reduction in quality of life for patients. The major effect is in the psychological area, with increased rates of emotional distress and impaired social functioning even in comparison with patients with major illnesses. However, most AN patients do report finding some benefit from their illness experience. Patients with facial weakness are at the greatest risk of a poorer psychological outcome. PMID- 19128649 TI - Voice-related quality of life in patients with benign vocal fold lesions. AB - OBJECTIVES: Efforts to assess the level of voice-related disability through application of written instruments are gaining wider acceptance in clinical practice. However, based on the questions posed, additional diagnostic information may be obtained. This study evaluated the potential extended utility of the Physical Functioning (PF) subscore of the Voice-Related Quality of Life (V RQOL) measurement instrument. Specifically, we examined whether the PF subscore could distinguish between the presence or absence of vocal fold lesions in patients who presented with dysphonia secondary to hyperfunctional voice disorders. A normative control database was also used for comparative purposes. DESIGN: Comparative analysis. SETTING: Academic tertiary care centre. METHODS: Forty adults with a hyperfunctional voice disorder. All had been seen for medical evaluation to confirm laryngeal status. MAIN OUTCOME MEASURES: V-RQOL total and domain scores. RESULTS: Although variability in V-RQOL scores was observed, differential profiles emerged from comparisons of participants who presented with vocal fold pathology and those who did not. Significantly lower PF and total V RQOL scores were observed for those with benign mass lesions of the vocal folds. Additionally, discriminant analysis of the data permitted the generation of a cutoff value for the PF subscore, which identified 80% of those with mass lesions. These data also permitted calculation of sensitivity, specificity, and positive predictive and negative predictive values. CONCLUSIONS: The findings suggest that the PF subscore may exhibit the capacity to distinguish dysphonic patients who present with a mass lesion of the vocal folds from those who do not within the context of a hyperfunctional voice disorder. Clinical implications and applications of the V-RQOL are discussed. PMID- 19128650 TI - Does routine preoperative imaging of parotid tumours affect surgical management decision making? AB - OBJECTIVE: To determine whether preoperative radiologic imaging of parotid tumours would alter the surgical approach to complete eradication of the parotid tumours. METHODS: A retrospective chart review of 173 patients with parotid tumours who underwent removal of parotid tumours at Mount Sinai Hospital in Toronto, Ontario, from 2000 to 2002. Each patient presented with a single, unilateral parotid tumour. Ninety-eight patients underwent preoperative radiologic imaging prior to definitive surgeries. The remaining 75 patients did not undergo preoperative imaging before surgery. RESULTS: Our study had shown that patients with superficial lobe parotid tumours did not require preoperative imaging as it did not affect the surgical management approach. On the contrary, for patients with parotid tumours clinically involving the deep lobe or parapharyngeal space, preoperative imaging often provides important additional information that may alter the surgical approach to eradication of parotid tumours. Moreover, patients with histopathologically benign tumours did not seem to benefit from preoperative imaging. On the other hand, patients with histopathologically malignant parotid tumours benefited from preoperative imaging as the additional information often altered the surgical management decision. CONCLUSION: Preoperative imaging should not be routinely ordered to investigate patients with parotid tumours. Patients with either deep parotid tumours or clinically suspicious tumours of malignancy would benefit from a preoperative radiologic investigation. The additional information from imaging may affect the surgical management decision. PMID- 19128651 TI - Thyroid gland in carcinoma of the larynx and hypopharynx: analysis of factors indicating thyroidectomy. AB - BACKGROUND: This prospective study analyzed the factors that could predict thyroid gland involvement in patients suffering from malignancy of the larynx and laryngopharynx. METHODS: Fifty patients underwent total laryngectomy with either ipsilateral hemithyroidectomy or total thyroidectomy in cases with subglottic extent more than 1.5 cm or in patients undergoing total laryngopharyngectomy for hypopharyngeal malignancy. The laryngectomy specimen and thyroid specimen were studied histologically with multiple serial sections. RESULTS: The majority of patients had glottic involvement followed by supraglottic and hypopharyngeal lesions. The thyroid gland was found to be enlarged and nodular in six patients. One of six patients had frank induration with gross extralaryngeal spread. Our study showed a 2% incidence of thyroid gland invasion by tumour. CONCLUSIONS: The presence of subglottic extension of more than 2 cm, cricoid cartilage invasion, and perithyroidal soft tissue involvement can be considered indicators for sacrificing the whole thyroid gland during surgery. PMID- 19128652 TI - Over-the-fence cochlear implantation: is it worthwhile? AB - OBJECTIVE: The objective of the study was to estimate the upper limit of speech perception for cochlear implant candidates. DESIGN: A retrospective chart review of more than 700 cochlear implant users was conducted. SETTING: All patients received their implant through the Quebec Cochlear Implant Program, which is a hospital- and rehabilitation centre-based program. METHODS: Charts from patients with preimplantation sentence recognition performance exceeding 40% were selected. Postoperative performance was compared with preoperative results. MAIN OUTCOME MEASURES: Sentence and word recognition was assessed with a multimedia auditory test battery. RESULTS: Comparison of pre- and postimplantation sentence recognition in the best aided condition shows some theoretical gain up to a 80 to 85% score for in-quiet sentence recognition tests before the surgery. Comparison of preimplantation sentence recognition in the best aided condition and postimplantation word recognition with the implant alone shows some gain up to a 60% score for in-quiet sentence recognition tests before the surgery. CONCLUSIONS: The speech perception selection criteria can confidently be expanded to 60% from a conservative perspective and even up to the 80% range if the use of a contralateral hearing aid is highly expected. PMID- 19128653 TI - Does extracorporeal shock wave lithotripsy cause hearing impairment? Evaluation by transient evoked otoacoustic emissions. AB - OBJECTIVE: To evaluate if multiple sessions of extracorporeal shock wave lithotripsy (ESWL) have any effect on the transient evoked otoacoustic emissions (TEOAEs). STUDY DESIGN: Prospective study. SETTING: Academic tertiary medical centre. METHODS: Thirty patients undergoing multiple sessions of ESWL for urinary stone disease were assessed using TEOAEs before and after ESWL. RESULTS: It was found that ESWL does not cause any significant changes in TEOAEs. CONCLUSIONS: ESWL is a safe procedure and does not cause any changes in TEOAEs. PMID- 19128654 TI - Ruptured thoracic aortic aneurysm: unusual cause of dysphagia. PMID- 19128655 TI - Isolated extramedullary plasmacytoma of the true vocal cord. PMID- 19128656 TI - Lipoma of the paranasal sinus: a rare but noticeable entity. PMID- 19128657 TI - Bilateral lateral saccular cyst. PMID- 19128658 TI - Malignant melanoma of the head and neck: presentation of three cases and review of the literature. PMID- 19128659 TI - Chondrosarcoma of the larynx: case report with a 14-year follow-up. PMID- 19128660 TI - Facial nerve neuroma associated with hemangioma of the geniculate ganglion: case report and review of the literature. PMID- 19128661 TI - Congenital oral heterotopic gastrointestinal cyst: case report and review of the literature. PMID- 19128662 TI - Eosinophilic angiocentric fibrosis. PMID- 19128664 TI - Modifications in nasal function and nitric oxide serum level in type 1 diabetes. AB - OBJECTIVE: In this study, we evaluated the modifications of nasal function in type 1 diabetes (insulin-dependent diabetes mellitus [IDDM]) by active anterior rhinomanometry (AAR) to understand if involvement of the nasal nervous system and microcirculation could be detected in nasal mucosa. METHOD: We studied 35 nonsmoking IDDM patients without diabetic complications, nasal pathology, or septal deviation. We measured serum levels of nitric oxide (NO) and nasal airway in three conditions: basal, supine, and after decongestion (phenylephrine hydrochloride 0.25 mg spray) by means of rhinomanometry, determining inspiratory total resistance and nasal airflow. The rhinomanometric results of the IDDM patients were compared with those of control normal subjects. In the IDDM patients, neuropathy was evaluated according to standardized procedures, including the vibration perception threshold test, cardiovascular autonomic tests, conduction velocity test, and fundoscopic examination. RESULTS: The NO serum level was significantly higher in IDDM patients (12.5 +/- 3.8) compared with normal controls (4.8 +/- 1.4). The AAR results showed that in IDDM patients, inspiratory total resistance in the basal (0.82 +/- 0.4 Pa/cm3) and supine (0.94 +/- 0.7 Pa/cm3) positions and after decongestion (0.59 +/- 0.2 Pa/cm3) were increased compared with the control group in three conditions (basal, 0.52 +/- 0.2 Pa/cm3; supine, 0.58 +/- 0.3 Pa/cm3; after decongestion, 0.48 +/- 0.2 Pa/cm3). After decongestion, there was a greater decrease in nasal resistance in diabetic patients than in normal subjects. CONCLUSION: Nasal function is involved in IDDM, rhinomanometry can also be considered an important test in the evaluation of this involvement in patients without other signs of diabetic neuropathy, and an increase in NO could partially explain these alterations. PMID- 19128665 TI - Nontuberculous mycobacterial cervicofacial adenitis in children: epidemiologic study. AB - OBJECTIVES: To determine whether there is an increase in the incidence of nontuberculous mycobacteria (NTM) adenitis at our tertiary care pediatric hospital and to determine whether there is an association between geographic location and prevalence of cases. Also, to investigate the role of nongeographic risk factors such as bacille Calmette-Guerin (BCG) vaccination in the incidence of NTM adenitis. METHOD: We conducted a retrospective chart review of 153 pediatric patients with NTM adenitis diagnosed between 1994 and 2004 and a prospective telephone survey of geographic and nongeographic risk factors for 82 patients. RESULTS: The annual incidence, calculated with the number of hospitalizations for that year as the denominator, was 25.1 in 100,000 during the 1994-1998 period and 107.4 in 100,000 for 1999-2004. A statistically significant increase in the incidence of NTM adenitis was observed after 1999, with an incidence ratio of 4.28 (95% confidence interval 2.84-6.65; p < .0000). We found a nonsignificant increase in incidence for cases in the suburbs compared with the city. The survey did not reveal any significant risk factors for NTM adenitis in our population. However, none of the affected children had had the BCG vaccination. CONCLUSION: There has been a significant increase in the incidence of NTM adenitis at our tertiary care centre, particularly since 1999. No clear risk factors could be identified to account for this increase, even though geographic location and discontinuation of BCG vaccination could be implicated. More studies are needed to confirm those hypotheses. PMID- 19128666 TI - Effects of helicobacter pylori eradication on chronic nonspecific pharyngeal symptoms. AB - BACKGROUND AND OBJECTIVES: Although its pathophysiology is not obvious, the discussion on chronic nonspecific pharyngeal symptoms encompasses a broad range of various factors, such as nasal obstruction, extraesophageal reflux disease, some of the chronic upper respiratory infections, and allergy. We found a high seroprevalence of Helicobacter pylori in a group of selected patients with no possible conditions responsible for complaints. To show a possible role of H. pylori infection in chronic pharyngeal complaints, we investigated symptomatic response following eradication therapy. PATIENTS AND METHODS: In a prospective cohort study, 72 subjects with H. pylori seropositivity had their symptoms measured by a questionnaire before and 3 months following H. pylori eradication therapy. The symptomatic response was compared between eradicated and noneradicated groups 3 months after eradication therapy. RESULTS: In 39 of 72 patients H. pylori was eradicated successfully. Mann-Whitney U analysis showed the pharyngeal symptom improvement rate, measured using the absolute difference in symptom score. For two major symptoms (chronic throat irritation, sore throat), it was significantly lower for H. pylori-noneradicated cases than for H. pylori-eradicated cases (p = .003 and p < .001, respectively). CONCLUSION: Resolution of the symptoms in the eradicated group suggests that H. pylori has a role in developing chronic nonspecific pharyngeal symptoms. These data may be important for future treatment strategies for this disease. PMID- 19128667 TI - Gelatinolytic activity of matrix metalloproteinases 2 and 9 in middle ear cholesteatoma. AB - BACKGROUND: Cholesteatoma of the middle ear or mastoid is a hyperproliferative disorder of keratinocytes characterized by a progressive bone erosion. Matrix metalloproteinase (MMP)-2 and MMP-9 gelatinases are endopeptidases targeting extracellular protein. Several studies examined the role of gelatinases in the pathogenesis of cholesteatoma, but the biologic mechanism by which cholesteatoma destroys the bone tissue remains unclear. OBJECTIVES: The aim of this study was to characterize the activity of MMP-2 and MMP-9 in human cholesteatoma and external auditory canal skin. METHODS: In the study, specimens of cholesteatoma and middle ear canal skin from 14 patients treated surgically at the Department of Otolaryngology were used. After two-step extraction of MMP-2 and MMP-9 from tissue samples, gelatinolytic activity was assessed with zymography. RESULTS: We noticed the augmentation of MMP-9 (p = .0001) and MMP-2 (p = .046) activity obtained from cholesteatoma in comparison with control skin. The MMP-9 active to latent ratio was significantly higher in cholesteatoma samples versus normal skin. CONCLUSION: The present study indicates that MMP-9 and, to a lesser degree, MMP-2 overexpression may be implicated in the molecular mechanisms of cholesteatoma invasion and bone destruction. PMID- 19128668 TI - Harmonic scalpel in pharyngolaryngectomy with radical neck dissection. AB - OBJECTIVE: The aim of this study was to verify the efficacy and applicability of the Ultracision Harmonic Scalpel (Ethicon Endo-Surgery, Cincinnati, OH) in pharyngolaryngectomy with radical neck dissection. DESIGN: This work evaluated the use of the Harmonic Scalpel in otolaryngology as a new and alternative method to overcome some complications of traditional surgery. SETTING: The study was conducted with 20 patients in the ENT Department of the University of Genoa (Italy) between January 1, 2005, and December 31, 2006. METHODS: A prospective, randomized study was undertaken on 20 pharyngolaryngectomies with radical neck dissection performed using a Harmonic Scalpel (10 patients) or traditional surgery (10 patients). MAIN OUTCOME MEASURES: The evaluation included operation time, intraoperative blood loss, postoperative seroma formation, and pattern of wound healing. RESULTS: In patients treated with Harmonic Scalpel, the mean operation time was significantly shorter, the blood loss was less, laryngeal and neck drainage on the first and second postoperative days was significantly smaller, and no postoperative complications were noted. CONCLUSIONS: The use of the Harmonic Scalpel during pharyngolaryngectomy and radical neck dissection led to diminished bleeding, shorter operative time, less seroma formation, and better wound healing in the postoperative period. PMID- 19128669 TI - Delayed sodium thiosulphate administration reduces cisplatin efficacy on mouse EMT6 tumour cells in vitro. AB - OBJECTIVE: To determine whether simultaneous and/or delayed administration of sodium thiosulphate (STS) affects the oncologic effect of cisplatin or cisdiaminedichloroplatinum (CDDP) in EMT6 tumour cells in vitro. SETTING: Cell biology research laboratory. METHODS: Clonogenic assays of EMT6 tumour cells with CDDP alone, CDDP plus simultaneous STS, and CDDP plus a 4-hour delay of STS were performed. Growth fractions under these three conditions were compared. RESULTS: Tumour growth was statistically significantly increased when CDDP and STS were administered compared with CDDP alone. There was no statistically significant difference between simultaneous and 4-hour delay of STS administration. We conclude that in EMT6 cells, either simultaneous administration or a 4-hour delay of STS administration significantly decreases CDDP efficacy. CONCLUSION: STS adversely affects CDDP's oncologic efficacy in EMT6 cell cultures in vitro. PMID- 19128670 TI - Incidence of adenoid cystic carcinoma in nova scotia: 30-year population-based epidemiologic study. AB - INTRODUCTION: Adenoid cystic carcinoma (ACC) of the head and neck is a rare malignancy of salivary gland origin. Few epidemiologic studies exist assessing the incidence of this tumour. The purpose of the current study was to determine the incidence of ACC of the head and neck in Nova Scotia. Furthermore, we assessed survival outcomes based on presentation sites. METHODS: Nova Scotia is an ideal geographic location in which to conduct retrospective case reviews as there are centralized tertiary care centres and minimal population migration. Furthermore, there is an externally and internally validated cancer registry housed in Cancer Care Nova Scotia. Records at Cancer Care Nova Scotia were retrospectively reviewed to identify all reported cases of ACC since 1979. Age adjusted incidence rates were calculated standardized to the 2001 Canadian population. Survival curves were plotted to allow assessment of overall survival. A log-rank test was used to compare survival curves for age (> 65 and < 65 years old) and presentation sites. RESULTS: One hundred thirty cases of ACC were identified, with 110 presenting as primary tumours in head and neck sites. The mean (standard error) age-adjusted incidence of ACC in Nova Scotia is 4.5 (0.6) cases per 1 000,000. The incidence did not change significantly over time. The most common site of presentation was the minor salivary glands (40%), followed by the submandibular gland (18.2%) and the parotid gland (17.3%). There was no significant difference for overall and disease-free survival between nonsalivary, major salivary, and minor salivary tumours (p = .97). Significant reductions in survival were found for those diagnosed at > 65 years of age (p < .014) and those with a sinus presentation (p < .005). Sixteen percent of all head and neck cancer patients experienced a distant metastasis of their primary tumour. CONCLUSION: ACC of the head and neck is a rare neoplastic condition that most commonly affects the minor and major salivary glands. Although short-term survival is high, nearly half of all patients will develop metastasis or die of complications of local recurrences. PMID- 19128671 TI - [Management of the papillary microcarcinoma of the thyroid gland]. AB - OBJECTIVES: To study the management and treatment of papillary microcarcinoma of the thyroid. MATERIAL AND METHODS: Retrospective study of 57 patients operated on their thyroid gland between 1995 and 2004. All patients had a frozen section of the specimen. Procedure varied according to histology results and presence of adenopathy. RESULTS: Procedure planned initially was a total thyroidectomy in 75% of patients (the other 25% were planned as hemi-thyroidectomy). Frozen section was positive in 46% of patients. We proceeded with 16 selective neck dissections (levels 2-3-4) and 10 bilateral "level 6" . Microcarcinomas were unique in 77% of cases. Extra-capsular spreading was noted in 39% of cases. Adjuvant Iodine 131 was administered to 36 patients (63%). Follow-up included clinic visits, ultra sound and Tg levels. With a follow-up of 3 to 12 years, no recurrence or metastasis has been identified. CONCLUSION: Treatment of papillary microcarcinomas of the thyroid is controversial. If surgery is chosen as treatment, the extent both regarding the thyroid itself and the cervical nodes is debated. Because of the excellent prognosis, some authors have recommended, when risk factors are absent, to limit the treatment to the thyroidectomy. Others, because of the risk of local recurrence and distal metastasis, have recommended a very aggressive approach. PMID- 19128672 TI - Lansoprazole in globus pharyngeus: double-blind, randomized, placebo-controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of once-daily lansoprazole in the typical globus pharyngeus patient, who presents with a normal head and neck examination and without other sinister otolaryngologic complaints. STUDY DESIGN: Prospective, randomized, double-blind, placebo-controlled trial. SETTING: Outpatient otolaryngology office practice. METHODS: After meeting the inclusion criteria, patients were randomized to a 3-month course of either treatment (30 mg lansoprazole daily) or placebo (lactose). All patients were given a medication diary and instructed to take the medication daily on waking. During the office visit, counseling regarding lifestyle modifications for gastroesophageal reflux disease was provided to all patients. MAIN OUTCOME MEASURE: The Glasgow-Edinburgh Throat Scale questionnaire was given prior to treatment and at 1- and 3-month intervals during treatment. RESULTS: Forty patients were enrolled between June 2005 and December 2006. Nineteen patients were randomized to the lansoprazole treatment arm, whereas 21 were enrolled in the placebo arm. Both groups were similar with regard to age, sex, and smoking history. The globus symptom score improved an average of 6.7 of 70 points in the treatment group and 2.9 points in the placebo group. There was no clinically or statistically significant difference between the two groups at any time point. CONCLUSION: The globus pharyngeus patient who presents with a normal head and neck examination and without sinister otolaryngologic complaints does not benefit from once-daily lansoprazole therapy. PMID- 19128673 TI - Ototoxic effects of single-dose versus 19-day daily-dose gentamicin. AB - INTRODUCTION: Gentamicin is one of the most extensively studied aminoglycoside antibiotics. The dogma of gentamicin ototoxicity theorizes that (1) the toxic effects of the drug are cumulative and dose dependent, despite clinical observations of ototoxicity after a single dose, and (2) gentamicin's ototoxic effects are irreversible, although clinicians have observed improvement in hearing over time. The objective of this study was to evaluate this basic dogma by examining the ototoxic differences between single-dose and 19-day daily dosing of gentamicin over a 60-day period. METHODS: Thirty-six C57 mice were randomly assigned to one of three treatment groups: (1) 19-day daily normal saline intraperitoneal injections (control; n = 10), (2) single-dose intraperitoneal 120 mg/kg gentamicin (n = 12), and (3) 19-day daily intraperitoneal 120 mg/kg gentamicin (n = 14). Pure-tone testing using auditory brainstem response was performed at frequencies of 6, 8, 12, 20, and 30 kHz. Hearing threshold was determined at each frequency by presenting stimuli from 90 dB to 5 dB using 10 dB decrements. Pure-tone testing was performed at days 1, 35, and 60 +/- 2 days. RESULTS: The results showed that hearing (1) improved between days 35 and 60 (p = .023) and (2) was not significantly different between a single dose versus 19 daily doses of gentamicin (p = .285). CONCLUSION: This study concurs with clinical observations that a single large dose of gentamicin may have ototoxic effects similar to those of multiple doses of gentamicin and that, over time, there is the potential for hearing recovery from gentamicin ototoxicity. PMID- 19128674 TI - Needs assessment of undergraduate education in otolaryngology among family medicine residents. AB - OBJECTIVES: To investigate the adequacy of otolaryngology teaching in undergraduate medical education (UME) and to determine the general level of comfort of family medicine residents in managing and assessing diseases affecting the ears, nose, and throat. DESIGN: Cross-sectional survey. METHODS: A sample of family medicine residents at the University of Western Ontario (N = 68) completed a questionnaire inquiring into their exposure to topics in otolaryngology during their UME. They were also asked to comment on their present comfort with knowledge and skills specific to otolaryngology. RESULTS: Family medicine residents suggest that there is generally little otolaryngology training in UME; 66.7% of respondents who received UME in Canada suggested that they received very little classroom instruction and 75.6% received very little clinical otolaryngology instruction. Residents identified specific otolaryngologic conditions with which they felt particularly uncomfortable. CONCLUSIONS: This study demonstrated that students receive very little exposure to otolaryngology in UME and highlighted specific conditions and procedures that family medicine residents lack confidence in managing and performing. PMID- 19128675 TI - Revision endoscopic sinus surgery: the St. Paul's Sinus Centre experience. AB - OBJECTIVE: To determine the most common findings in patients undergoing revision endoscopic sinus surgery (ESS) presenting to a tertiary rhinology centre. It is our aim that by identifying these findings, the common pitfalls in primary ESS can be avoided to prevent the necessity for revision ESS. METHODS: The findings of 73 cases of revision ESS from July 2006 to March 2007 presenting to the St. Paul's Sinus Centre were recorded and then presented. RESULTS: There are many common findings at revision ESS, including residual uncinate process, persistent septal deviation, nonphysiologic maxillary antrostomies, incomplete ethmoidectomy, and partial or total resection of the middle and superior turbinate, resulting in the formation of the "uniturbinate." CONCLUSION: Many common findings in revision ESS can be avoided with proper primary surgery. PMID- 19128676 TI - Significance of laryngeal edema after partial laryngectomy and radiotherapy in supraglottic cancer. AB - OBJECTIVE: To evaluate the impact of the presence of laryngeal edema on outcome and the other potential prognostic factors in patients with supraglottic larynx carcinoma treated with radiotherapy after partial laryngectomy. DESIGN: A retrospective analysis. SETTING: Cerrahpasa Medical Faculty, Istanbul University. METHODS: Seventy-nine patients with supraglottic carcinoma of the larynx were treated with radiotherapy after partial laryngectomy between January 1980 and July 2003. Neck dissection was not performed in 46 patients. The median follow-up time was 64 months. MAIN OUTCOME MEASURES: The effect of laryngeal edema grade on local control, disease-free, and actuarial survival rates. RESULTS: The 5-year locoregional progression-free and overall survival rates were 86% and 75%, respectively. In univariate analysis, histologically positive neck disease reduced regional (p = .0045) and disease-free survival (p = .01). Patients with edema grade III-IV had lower local control (p = .0004), disease-free (p = .0034), and actuarial survival (p = .0041) rates. In the multivariate analysis, a significant negative association of laryngeal edema with local control (p = .012), disease-free survival (p = .002), and actuarial survival (p = .003) was found. Nodal status was a significant prognostic factor for disease-free survival (p = .027). Grade III-IV laryngeal edema was observed in 17 patients. Owing to laryngeal edema, tracheostomy dependence and total laryngectomy were required in three patients and one patient, respectively. CONCLUSION: Radiotherapy after partial laryngectomy can be performed in patients with poor prognostic factors with reasonable complication rates. However, in the presence of grade III-IV laryngeal edema, tumour recurrence should be suspected, and these patients have to be managed with close follow-up and further evaluation to improve outcome. PMID- 19128677 TI - IIb or not IIb: oncologic role of submuscular recess inclusion in selective neck dissections. AB - OBJECTIVE: Selective neck dissections (SNDs) can yield outcomes that are oncologically similar to radical dissections but with less morbidity. The rate of metastasis to level IIb is not clear, and its dissection involves cranial nerve XI traction and devascularization, causing much morbidity. Our study examined the prevalence and predictors of cancer within the submuscular recess (SMR). METHODS: All SNDs performed by the authors were prospectively included from July 1, 2002, to March 31, 2006. Level IIb was sent as a distinct specimen. RESULTS: One hundred fifty-two dissections were performed. Only 12 (7.9%) were node positive in IIb. The SMR contained diseased nodes in 12.2% of N+ necks and 3.0% of N0 necks (p = .04); 23.1% (3 of 13) of laryngeal (1 of 8) and hypopharyngeal (2 of 5) tumours were node positive in IIb versus 6.4% in the oral cavity (p = .07) and 5.3% in oropharyngeal lesions. CONCLUSIONS: This is one of the largest prospective studies examining the role of level IIb dissection. It suggests that level IIb dissection might be unnecessary, especially in an N0 neck. PMID- 19128678 TI - Effect of 18F-fluorodeoxyglucose positron emission tomography on the management of patients with head and neck squamous cell carcinoma. AB - OBJECTIVE: Positron emission tomography (PET) has become a commonly used imaging modality for initial staging and for surveillance of patients with head and neck cancer. It is reported to be more sensitive and specific than computed tomography and magnetic resonance imaging in the detection of primary and metastatic tumours. The goal was to determine whether the information provided by PET alters the management of patients with head and neck squamous cell carcinoma. MATERIALS AND METHODS: The records of patients who underwent PET as part of the initial staging evaluation or for posttreatment surveillance were reviewed. RESULTS: PET was obtained in 29 patients as part of the initial staging evaluation and in 18 patients for posttreatment surveillance. In some patients, PET provided information beyond the information obtained from the routine workup and evaluation. This additional information altered the management of six patients (13%). Two of these patients were in the initial staging group and four patients were in the surveillance group. The management changes prompted by these results were unnecessary or erroneous in three of these six patients. CONCLUSIONS: 18F Fluorodeoxyglucose (FDG)-PET allows assessment of primary tumours, regional metastases, and distant metastases for the purpose of initial staging and for surveillance in previously treated patients. The information provided by the FDG PET prompted further workup or altered the treatment plan of 13% of the patients. These changes in management were sometimes unnecessary. PMID- 19128679 TI - Differences in Ototoxicity across Species. AB - OBJECTIVE: To illustrate some differences between humans and rodents in the dose effect relationships for two ototoxic drugs. STUDY DESIGN: Controlled animal study using typical research regimens for gentamicin and cisplatin compared with human data from the clinical literature. METHODS: Auditory brainstem response testing was carried out over months in two groups of animals exposed to typical dose regimens for ototoxic drugs. In the first group, 30 guinea pigs received either 3 or 6 mg/kg of cisplatin on alternate days for 5 days (total dose 15 or 30 mg/kg). In the second group, 24 C57 mice received saline or 19 daily doses of gentamicin 120 mg/kg. The findings in rodents were contrasted with human toxicity in the literature. RESULTS: Cisplatin increased click thresholds (32 +/- 27 dB) in guinea pigs. Doses of 15 mg/kg caused less hearing loss than 30 mg/kg, but the higher dose was associated with greater mortality owing to renal insufficiency. These findings are comparable with expectations of similar doses of cisplatin in humans. In contrast, gentamicin produced less hearing loss in mice, although the dose employed was well above the lethal dose for humans. CONCLUSIONS: Ototoxic doses of cisplatin in guinea pigs are similar to those of humans, but C57 mice appear to be highly resistant to gentamicin-induced hearing loss compared to humans. Animal models of ototoxicity need to be considered carefully in translational research. PMID- 19128680 TI - Vestibular evoked myogenic potential eliciting in normal subjects: comparison of four different methods. AB - OBJECTIVE: Vestibular evoked myogenic potential (VEMP) recording is a new method for testing the otolith receptors and vestibulospinal pathways. The aim of this study was to evaluate the characteristics of VEMP using four different techniques to find reasons to prefer one type of recording over the others. MATERIAL AND METHODS: Twenty healthy persons, 10 males and 10 females with ages ranging from 20 to 57 years (mean age 41 years), were enrolled in this study. Eliciting of VEMPs by using monaural or binaural acoustic stimulation and unilateral or bilateral SCM contraction was evaluated; 105 dB NHL acoustic stimulation consisting of 145 dB rarefaction clicks was applied. Latencies of p13, n23, n34, p44 peaks; amplitudes p13-n23 and n34-p44; and interaural amplitude differences (IADs) were assessed. RESULTS: All four methods elicited constant and evident waveforms. The reliability coefficients of amplitudes were high for all four methods and for both waves. However, the higher scores of reliability appeared for the monaural-ipsilateral recording. The results indicated no statistically significant difference between the right and left sides for all four types of VEMP eliciting. No correlation was found between IAD13-23 and IAD34-44 for all four methods. Statistically significant differences were found only for n23 latency among the four methods. CONCLUSIONS: Although no evidence to reject or strongly favour a specific method was found, the monaural-ipsilateral recording was associated with some advantages. PMID- 19128681 TI - 18F-fluorodeoxyglucose positron emission tomography-computed tomography for suspected recurrent papillary thyroid cancer: early experience at Sunnybrook Health Sciences Centre. AB - OBJECTIVES: To report the initial experience with combined 18F-fluorodeoxyglucose positron emission tomography (FDG PET)/computed tomography (CT) imaging for suspected recurrent papillary differentiated thyroid cancer (DTC) at Sunnybrook Health Sciences Centre (SHSC), Toronto. DESIGN: Single institution retrospective study. METHODS: Consecutive patients from SHSC who underwent FDG PET/CT imaging for suspected recurrent DTC over a period of 2.5 years were identified and their charts reviewed. MAIN OUTCOME MEASURE: Qualitative appraisal of FDG PET/CT imaging in suspected recurrent DTC. RESULTS: Sixteen patients (14F, 2M) were identified accounting for 17 FDG PET/CT scans. Three scans (18%) in 3 different patients were reported as suspicious for recurrent disease in the neck (1-3 lesions) and were considered "positive". All were subsequently confirmed pathologically (4-13 positive lymph nodes post operatively). Prior conventional imaging was abnormal in two patients. Two patients had an elevated non-stimulated thyroglobulin (TG) < 10 ng/mL (4.9 and 9.4). The remaining patient had a TG < 0.3 ng/mL but was anti-TG antibody positive (84 IUx10-3/L). With a median follow up of 15 months (range 7-36) there were no false positive or negative scans. Incidental pathology (breast cancer, large bowel polyps) was identified on a further 2/17 scans (12%). CONCLUSIONS: FDG PET/CT imaging is able to detect recurrent DTC in patients with low TG levels. It can complement conventional imaging findings and exclude distant FDG-avid metastases prior to surgery. It may underestimate the number of positive lymph nodes in the neck. Occult pathology may be identified with whole body FDG PET/CT. PMID- 19128682 TI - Evaluation of cochlear function in normal-hearing young adults exposed to MP3 player noise by analyzing transient evoked otoacoustic emissions and distortion products. AB - OBJECTIVE: Exposure to recreational noise may cause injuries to the inner ear, and transient evoked (TEOAEs) and distortion product otoacoustic emissions (DPOAEs) may identify these cochlear alterations. The goal of this study was to evaluate TEOAEs and DPOAEs as a method to diagnose early cochlear alterations in young adults exposed to MP3 player noise. MATERIAL AND METHODS: We performed a prospective study of the cochlear function in normal-hearing MP3 player users by analyzing TEOAE and DPOAE incidence, amplitude, and spectral content. We gathered a sample of 40 ears from patients between 19 and 29 years old (mean age 24.09 years, SD 3.9 years). We compared the results with those of a control group of 232 ears not exposed to MP3 noise from patients aged 18 to 32 years (mean age 23.35 years, SD 2.7 years). Fifty percent of ears were from females and 50% were from males. RESULTS: Subjects who had used MP3 players for most years and for more hours each week exhibited a reduction in TEOAE and DPOAE incidence and amplitudes and an increase in DPOAE thresholds. TEOAEs showed a statistically significant lower incidence and amplitudes for normal-hearing subjects using MP3 players at frequencies of 2000, 3000, and 4000 Hz. DPOAE incidence was lower at 700, 1000, 1500, and 2000 Hz; the amplitudes were lower at frequencies between 1500 and 6000 Hz; and the thresholds were higher for all frequency bands, statistically significant at frequencies from 1500 to 6000 Hz, p < .05. CONCLUSIONS: Cochlear impairment caused by MP3 player noise exposure may be detectable by analyzing TEOAEs and DPOAEs before the impairment becomes clinically apparent. PMID- 19128683 TI - Nasolacrimal duct stenosis in children on the level of the Hassner valve: resolving the stenosis with a polypropylene thread knot. PMID- 19128684 TI - Eagle syndrome: teaching the intraoral surgical approach with a 30 degrees endoscope. PMID- 19128685 TI - Microvascular changes in radiation-induced oral mucositis. AB - BACKGROUND: Mucositis is one of the most debilitating side effects of head and neck cancer therapy and is currently believed to arise from an inflammatory cascade leading to cellular damage. However, no effective treatment has been identified despite extensive attempts with anti-inflammatory medications. OBJECTIVE: To compare real-time microvascular inflammatory changes with oral mucositis levels in patients undergoing radiotherapy or chemoradiotherapy for head and neck tumours. DESIGN: Prospective, longitudinal, cohort, observational study. SETTING: Regional cancer program. METHODS: Twenty patients with head and neck tumours were assessed on a weekly basis throughout the course of radiotherapy. Levels of mucositis were graded objectively using the Oral Mucositis Assessment Scale and subjectively using a patient symptom questionnaire. Video imaging of the sublingual microcirculation was obtained using orthogonal polarized spectral imaging to quantify inflammatory markers such as microcirculatory velocity, white blood cell margination, and extravasation. RESULTS: Despite very high levels of objective and subjective mucositis, inflammatory changes were not present in the microcirculation. CONCLUSIONS: Typical microvascular inflammatory changes are not demonstrated in radiation induced mucositis. These findings contradict the currently proposed mechanism of mucosal damage and may therefore have important implications in the development of novel therapeutic interventions. PMID- 19128686 TI - Head and neck osteosarcoma in adults: the province of alberta experience over 26 years. AB - OBJECTIVE: The goal of this study was to describe the incidence, treatment, and outcomes of osteosarcomas in the head and neck over 26 years in the province of Alberta, Canada. DESIGN: Retrospective analysis of 14 patients identified and treated in Alberta between 1974 and 1999. Overall and disease-specific survival were the most important outcomes of interest. RESULTS: The average age was 43 years. The maxilla was the most common primary site. The majority of the patients received surgical treatment in combination with adjuvant chemotherapy. Disease specific survival was 42% at 2 years and 30.1% at 5 years. Four patients developed osteosarcomas in the field of previous radiation. CONCLUSIONS: Osteosarcoma of the head and neck is very aggressive. In our series, mostly high grade malignancy affected a middle-aged population. Surgical resection is difficult owing to anatomic restraints, and adjuvant chemotherapy should be considered in most patients. The series is notable for the poor outcome, with a 5 year actuarial survival of 30%. This may reflect the lack of a standardized treatment protocol. PMID- 19128687 TI - Conservative management of warthin tumour. AB - BACKGROUND: Warthin tumour (WT) is usually treated by surgery to establish a histologic diagnosis. Conservative management is considered appropriate for patients with significant comorbidities, making anesthesia high risk. Preoperative fine-needle aspiration cytology (FNAC) is useful in diagnosing WT, making conservative management an option. In our institution, patients with an FNAC diagnostic of WT have the option of conservative management. METHOD: Retrospective review of cases of WT over 10 years (1996-2006) to study the demographics of patients treated conservatively and surgically and to establish the accuracy of FNAC. RESULTS: Eighty-six cases of WT were identified during the study period for which FNAC was diagnostic, of which 58 cases (67.4%) were treated conservatively and 28 cases (32.6%) were treated by superficial parotidectomy. A further seven cases of WT were diagnosed on histology as the corresponding FNAC was incorrect in two cases and nondiagnostic in five cases. With regard to WT, FNAC in our unit had a sensitivity of 80% and a specificity of 100%. Females accounted for 54.7% of cases, which is much higher than previously reported and likely reflects the high prevalence of smoking among females in Liverpool. CONCLUSION: Conservative management for WT on the basis of a diagnostic FNAC was employed in 67.4% of cases. This included young patients who preferred to avoid surgery. Conservative management is a viable option if reliable cytologic reporting is available. PMID- 19128688 TI - Profound hearing loss associated with oxycodone-acetaminophen abuse. PMID- 19128689 TI - Head and neck reconstruction with an extended latissimus dorsi pedicled flap: report of three cases. PMID- 19128690 TI - Sneezing-related orbital emphysema and pneumocephalus treated with transnasal endoscopic surgery. PMID- 19128691 TI - Isolated asymmetric palatal palsy owing to Mycoplasma pneumoniae infection. PMID- 19128692 TI - Endoscopic management of malignant inverted papilloma of the nasopharynx. PMID- 19128693 TI - Rare case of paratracheal hydatid cyst. PMID- 19128694 TI - Chronic sinusitis and alpha1-antitrypsin deficiency: potential role for protease in rhinosinusitis? PMID- 19128695 TI - Thyroid cartilage metastasis from the lung. PMID- 19128696 TI - Nodular fasciitis of the maxillofacial region in a pediatric patient. PMID- 19128697 TI - Tenacious nasal polyposis in cystic fibrosis patients subsequent to bilateral lung transplantation. PMID- 19128698 TI - Tumour-to-tumour metastasis from a carcinoma of the breast to a pleomorphic adenoma of the parotid gland. PMID- 19128699 TI - Cross-sectional survey of hearing impairment and ear disease in Uganda. AB - OBJECTIVE: To determine the prevalence and causes of disabling hearing loss in adults and children in Uganda. STUDY DESIGN: Cross-sectional survey of ear disease and hearing impairment. SETTING: A random cluster sample design of the population from the Masindi district of Uganda following the World Health Organization (WHO) guidelines, using a modified version of the WHO Ear Disease Survey Protocol. MAIN OUTCOME MEASURE: The prevalence of disabling hearing impairment using the WHO definitions (excluding 0.5 kHz owing to high background noise levels). RESULTS: In the study, 6041 participants were enrolled and underwent audiometric evaluation and an ear examination. The prevalence of disabling hearing impairment was 11.7% in adults and 10.2% in children. A further 2.3% of children in whom thresholds could not be measured were deemed to have significant hearing loss based on screening questions and/or sound-field stimuli. Correctable causes such as dry perforations, cerumen impaction, and chronic suppurative otitis media resulted in disabling hearing loss in 17% of adult subjects and 41% of children. Preventable hearing loss, such as meningitis and noise-induced hearing loss, was present in a further significant percentage of subjects. CONCLUSIONS: Ear disease and hearing impairment were found to be important health problems in the Ugandan population. Preventable ear disease is a major cause of hearing loss in the population. It is hoped that the findings of this study will draw attention to the problem in Uganda and will lead to proper allocation of resources for the prevention and treatment of hearing loss and ear disease. PMID- 19128700 TI - Retrospective comparison of an endoscopic assisted versus a purely endoscopic approach to sellar tumour resection. AB - INTRODUCTION: The purely endoscopic approach to sellar tumors is thought to offer improved visualization and decreased complications compared to the endoscopic assisted approach. METHODS: This retrospective study was approved by the Research Ethics Board (REB) of McMaster University. Seventy-one consecutive patients who underwent sellar surgery at our institution from January 2003 to October 2005 were reviewed. 40 patients underwent an endoscopic assisted (microscope) approach, while 31 underwent a purely endoscopic approach. Populations were compared for differences in demographics, OR times, outcome measures, and complication rates. RESULTS: The groups were demographically equivalent. The pathology was similar across both groups. Both groups had a reduction of volume and invasion post operatively. There was a trend toward greater reduction in tumour volume and a significant decrease in tumour invasion in the purely endoscopic group. Both groups demonstrated similar improvements in visual fields. The purely endoscopic group had an improvement of 0.33 axes (p<0.05) in pituitary function. Overall, complications were reduced in the purely endoscopic group. A trend toward shorter operative times (128 minutes vs 135 minutes) and length of hospital stay (3.6 days (p<0.05) vs 5.4 days) in the purely endoscopic group. CONCLUSION: Our purely endoscopic group demonstrated a trend, as skill and confidence increased so did the size and invasiveness of the tumours being resected. Overall, the purely endoscopic technique showed improved total tumour resection, improved pituitary function, similar visual field outcomes, less post operative complications, decreased operative times and reduced hospital stay when compared to the endoscopic assisted technique. PMID- 19128701 TI - Meta-analysis: prophylactic drainage and bleeding complications in thyroid surgery. AB - OBJECTIVE: To conduct a comprehensive systematic review and high-quality meta analysis to determine whether prophylactic drain placement reduces adverse bleeding events in thyroid surgery. DATA SOURCES: MEDLINE (OVID and PubMed), CENTRAL, CDSR, ACP Journal Club, DARE, EMBASE, PREMEDLINE, OLDMEDLINE, CINAHL, BIOSIS Previews, LILACS, KOREAMED, SAMED, IndMED, SIGLE, ScienceDirect, and INGENTACONNECT. REVIEW METHODS: Studies for evaluation included all prospective trials assessing the use of drainage in thyroid surgery. We excluded case studies, retrospective studies, reviews, and studies that had a "selective" method of postoperative drainage that was not defined or was based on surgeon preference. Search strategies were broad and based on Cochrane Collaboration search filters. There was no language restriction. Article selection was conducted by two independent reviewers under QUORUM guidelines. RESULTS: Four hundred sixty-two articles were identified by the search strategy used, and 16 articles were included in the final review. Ten studies were randomized controlled trials, with 8 used for quantitative meta-analysis. No study showed a statistically significant benefit or harm with drain use. Meta-analysis of data estimated an odds ratio of 1.47 for reoperation for bleeding and 0.88 for visible hematoma for suction drains versus no drains. The results were not statistically significant, and 95% confidence intervals were wide. CONCLUSION: The literature has insufficient evidence to recommend routine drainage in thyroid surgery. It is possible that drains may increase the risk of reoperation for bleeding, although the data are not statistically significant. If there is a benefit to drainage, absolute risk reductions of bleeding outcomes may not warrant routine use. PMID- 19128702 TI - Penetrating and blast ear trauma: 7-year review of two pediatric practices. AB - OBJECTIVE: To review our experience with ear trauma in children. DESIGN/METHODS: Retrospective review of two practices from 2000 to 2007. SETTING: Pediatric tertiary care hospital. MAIN OUTCOME MEASURES: Micro-otoscopic findings and audiologic data. RESULTS: There were 18 cases of penetrating ear trauma (PET) and 6 cases of blast ear trauma (BET). The average age of the children with PET was 5 years. Fifteen of the 18 cases involved cotton-tipped applicators (CTAs); 8 patients had tympanic membrane perforations from CTA use. Six of the perforations healed spontaneously, and the other two patients were lost to follow-up. For 15 of the 18 PET patients, audiograms were available, and all eventually returned to normal. The average age of the six patients with BET was 11 years. BET caused five tympanic membrane perforations, all of which healed spontaneously. Three patients had audiograms, which were normal. The other three were lost to follow up. None of the patients had vertigo, nystagmus, facial weakness, or cerebrospinal fluid otorrhea; this factored into our nonsurgical approach. CONCLUSIONS: PET and BET in children are underreported. PET usually involves CTA and occurs in younger children than does BET. Urgent surgical intervention is not indicated unless a child presents with neuro-otologic signs or symptoms. CTA avoidance should be taught systematically to the public. PMID- 19128703 TI - Radiofrequency ablation of the tongue base in obstructive sleep apnea: rapid and effective technique using low-temperature radiofrequency molecular disassociation (coblation) for management of retroglossal obstruction. PMID- 19128704 TI - Case Series: raising awareness about Kikuchi-Fujimoto disease among otolaryngologists: is it linked to systemic lupus erythematosus? AB - OBJECTIVES: Kikuchi-Fujimoto disease (KFD) is a rare, benign, self-limiting condition occasionally associated with systemic lupus erythematosus (SLE). Although rising in incidence, nonspecific symptoms make diagnosis difficult. We present a small case series and review the literature to increase awareness of this condition among otolaryngologists. METHODS: Nine cases of KFD were diagnosed between 2003 and 2006 at the McGill University Health Care institution by either excisional or fine-needle biopsy. Clinical and histologic features were reviewed in the context of a comprehensive analysis of the KFD literature. RESULTS: Seven of nine cases are female, with a mean age of 27.2 years (range 18-35 years). Only one patient had systemic SLE predating KFD. Our results are comparable to those in the current literature regarding the association with SLE. CONCLUSIONS: We present the first Canadian small case series and extensive review of the literature to highlight the clinical and pathologic features of this disease and its association with SLE. PMID- 19128705 TI - Small cell carcinoma of the head and neck: a single institution's experience and review of the literature. AB - OBJECTIVES: It is well known that small cell carcinoma (SmCC) arising at extrapulmonary sites leads to a poor prognosis for patients. Owing to the infrequency of primary SmCC of the head and neck, it is very unlikely that any large, controlled study will ever be performed. For this reason, recommendations for the treatment of SmCC arising in the head and neck are based primarily on retrospective data from various small case series. PATIENTS AND METHODS: We report five patients with SmCC of the head and neck seen between 2001 and 2007. In two patients, the SmCCs are in the larynx; in another two patients, in the sinonasal region; and in one patient, in the tonsil. RESULTS: Four patients accepted concurrent chemoradiation therapy (CCRT). Their outcomes are as follows: two patients died, one has adrenal and bone metastasis but is still alive, and one shows no evidence of disease after treatment. One patient refused CCRT and died within 6 months. CONCLUSIONS: The prognosis for patients with primary SmCC of the head and neck is generally poor. Treatment most often involves a combination of radiotherapy and chemotherapy, and hope for improved outcomes hinges principally on the development of improved chemotherapies and other systemic treatments. PMID- 19128706 TI - Microscopic transtracheal repair of migrating tracheoesophageal fistula. AB - OBJECTIVE: The repair of a persistently leaking migrating tracheoesophageal fistula (TEF) represents a particular challenge owing to the low site of the fistula down to the tracheoesophageal septum (TES). A simple microscopic approach to repair a migrating TEF is described. DESIGN: A description of five cases of migrating TEF. The repair technique and surgical outcome are described in detail. SETTING: Tertiary care referral hospital. METHODS: Excision of the fistula tract was done under local anesthesia and microscopic vision using microlaryngoscopic instruments followed by one-layer repair without soft tissue interposition. This technique was used in one patient with a leaking migrating TEF when planned dissection through the TES was abandoned. Subsequently, the technique was employed in four other patients with a similar TEF. MAIN OUTCOME MEASURES: Evidence of complete closure of the fistula was assessed clinically 1 week postoperatively. This was followed by methylene blue and Gastrografin swallowing tests. The methylene blue test was repeated after 6 months to exclude recurrence of the fistula and confirm persistent closure. RESULT: Complete closure of the fistulae was achieved when assessed clinically and by methylene blue and Gastrografin tests. All patients were discharged on a normal diet. Stable closure was confirmed by the methylene blue test after 6 months. The microlaryngoscopic instruments and surgical microscope have greatly facilitated access and dissection of the migrating fistula with minimum soft tissue loss. CONCLUSION: The described technique is simple, relatively safe, and reproducible for closure of a small migrating TEF. It can also be used to repair small, nonmigrating TEF. PMID- 19128707 TI - The better performance status, the better outcome: laryngeal carcinoma treated with definitive radiotherapy. AB - PURPOSE: To evaluate the prognostic factors affecting locoregional control (LRC) and overall survival (OS) of patients with laryngeal carcinoma who were not candidates for surgical treatment due to tumour or host factors but were treated with definitive radiotherapy (RT). PATIENTS AND METHODS: Sixty-three consecutive patients, treated with definitive RT between 1999 and 2005, were retrospectively analyzed. All patients had histologically proven squamous cell carcinomas of the larynx. The median age was 62 years (range 43-83 years). Follow-up ranged from 22 days to 68 months (median 32 months). RESULTS: The LRC rates at 2 and 5 years were 70% and 48%. The 2- and 5-year OS rates were 65% and 40%. No statistically significant relationship was found between World Health Organization performance status score (WHO PS) and age (p=.21), tumour site (p=.42), overall stage (p=.11), T stage (p=.19), and N stage (p=.69). Multivariate analyses showed that a WHO PS score>or=2 (p<.0001) and RT treatment time>or=50 days (p=.0172) significantly decreased LRC. Moreover, a WHO PS score>or=2 (p<.0001), RT treatment time>or=50 days (p=.0138), and RT dose<66 Gy (p=.04) were significantly negative prognostic factors on OS. CONCLUSION: Definitive RT, in patients with early- and more advanced-stage squamous cell carcinoma of the larynx, is an important treatment option. It is clear that patients with good pretreatment PS would get better results from definitive RT. PMID- 19128708 TI - Sudden sensorineural hearing loss: have we got a cure? AB - OBJECTIVE: To determine the efficacy of the shotgun regimen and to assess the various prognostic variables affecting recovery in patients with sudden sensorineural hearing loss (SSNHL). MATERIAL AND METHODS: A prospective study of 42 patients with SSNHL was done. The selected patients received oral prednisolone at a dose of 1 mg/kg body weight tapered over 2 weeks and percutaneous application of 10 mg nitroglycerine for 1 week. RESULTS: There was a clinically significant improvement in hearing in 36.2% of the patients. A significant association was seen between the duration of hearing loss before seeking treatment and the recovery. Younger patients showed better improvement. The severity of hearing loss at presentation had no influence on the recovery. CONCLUSIONS: This regimen was found to be efficacious in the treatment of SSNHL. The single most important factor that influenced the recovery was the duration of hearing loss before seeking treatment. PMID- 19128709 TI - Refractory choanal atresia: what makes a child susceptible? The great Ormond Street Hospital experience. AB - OBJECTIVE: An outcome analysis of factors that predispose patients to require multiple surgical procedures for choanal atresia repair. DESIGN: Retrospective case note review of choanal atresia patients identified from a prospectively collected database at Great Ormond Street Hospital for Children, London, between 1992 and 2005. SETTING: Specialist centre in pediatric otolaryngology. METHODS: All patients underwent atresia correction by a transnasal approach under endoscopic guidance using a 120 degrees Hopkins rod telescope to visualize the posterior choanae from the nasopharynx. The atretic plate was first perforated using urethral sounds. The posterior choanal opening was subsequently enlarged using the microdebrider drill. Portex endotracheal tubes were used as stents in selected cases. RESULTS: Twenty children (9 male, 11 female) were identified who underwent multiple surgical procedures for restenosis following choanal atresia repair during the study period. The number of procedures per patient ranged from 6 to 42. Approximately half of the patients had other associated major anomalies. CONCLUSIONS: Following primary endoscopic transnasal repair, 9.8% of the patients were considered to have refractory choanal atresia, requiring six or more surgical procedures. Male gender, bilateral disease, associated congenital anomalies, low birth weight, and small stent size are potential risk factors for restenosis of choanal atresia. There was no obvious relationship between the duration of stent placement and restenosis. PMID- 19128710 TI - Safety of budesonide in saline sinonasal irrigations in the management of chronic rhinosinusitis with polyposis: lack of significant adrenal suppression. AB - OBJECTIVE: To evaluate the potential for hypothalamic-pituitary-adrenal (HPA) axis suppression by budesonide nasal irrigations in the treatment of refractory chronic rhinosinusitis with polyposis (CRSwP). STUDY DESIGN: Retrospective, descriptive review of patient charts. SETTING: Tertiary care rhinology practice in an academic teaching hospital. PATIENTS: Eighteen adult subjects with CRSwP refractory to conservative medical therapy. METHODS: The charts of consecutive patients identified as being treated with topical budesonide in saline for nasal irrigation from January to October 2006 were reviewed. In all cases, pre- and posttreatment morning cortisol levels had been measured following at least 8 weeks of uninterrupted therapy. In addition, a subset of patients who continued therapy longer than 8 weeks had undergone the more sensitive adrenocorticotropic hormone (ACTH) stimulation test. RESULTS: All pre- and posttreatment morning cortisol levels were within the normal range. For an 8-week treatment period, there was no evidence of HPA axis suppression (p=.4171). For patients who continued treatment beyond 8 weeks, ACTH stimulation did not detect HPA axis suppression. Furthermore, there were no issues with compliance or acceptability, nor were any adverse side effects reported. CONCLUSION: Budesonide in saline sinonasal irrigation for the treatment of refractory CRSwP does not cause HPA axis suppression. The efficacy of this higher dose of steroid delivered locally would benefit from further study. PMID- 19128711 TI - Comparison of rapid rhino and merocel nasal packs in endonasal septal surgery. AB - OBJECTIVE: Our study compared Rapid Rhino (RR; Applied Therapeutic Ltd, Leicestershire, UK) and Merocel (Medtronic Xomed, Jacksonville, FL) nasal packing materials in endonasal septoplasty surgery in terms of postoperative patient comfort and reactionary bleeding. PATIENTS AND METHODS: Fifty-one patients underwent endonasal septoplasty. One nasal cavity was packed with Merocel packs and the RR pack was used for the other side. The pain and nasal fullness levels on each side were studied at 1 and 6 hours postoperatively. Pain level was also studied during the removal of the nasal packs on the second day. These levels were scored on the basis of a visual analogue scale (VAS) between 0 and 10. Reactionary bleeding after nasal pack removal was also recorded. The Wilcoxon test was used for statistical analysis of the VAS scores, and the McNemar test was used for comparison of bleeding levels after pack removal. A p value<.05 was considered to be statistically significant. RESULTS: Minimal bleeding was noted in 15 patients following Merocel pack removal; however, no bleeding was seen following RR pack removal. Comparison of the VAS scores of pain and nasal fullness of each nasal pack at 1 and 6 hours postoperatively showed a statistically significant difference (p<.05). Similarly, comparison of the pain level for pack removal and bleeding after removal on the second postoperative day demonstrated a statistically significant difference (p<.05 and p=.001, respectively). CONCLUSION: RR is more tolerable by patients than Merocel after septoplasty surgery. RR also has some advantages, namely, easy pack removal and less reactionary bleeding. PMID- 19128712 TI - Risk factors for recurrence of benign paroxysmal positional vertigo. AB - OBJECTIVE: To assess the results of treatment for a first episode of benign paroxysmal positional vertigo (BPPV) and risk factors for recurrence. STUDY DESIGN: Retrospective chart review of 148 BPPV patients at a tertiary care referral centre. MATERIALS AND METHODS: The canalith repositioning procedure (CRP) was performed until vertigo and nystagmus were resolved. Using multivariate logistic regression, we estimated the relationship between the rate of BPPV recurrence within 1 year and various risk factors for BPPV. RESULTS: Among the 145 patients after successful manoeuvre or spontaneous resolution, BPPV recurred in 31 patients (21%) within 1 year. Patients with recurrent BPPV in the posterior semicircular canal required more CRP sessions and a longer period. The rate of BPPV recurrence in patients with endolymphatic hydrops (75%) was significantly higher than in those without (odds ratio 17; 95% confidence interval 2.9-101). Both the first episode of BPPV and recurrent BPPV affected the same ear associated with endolymphatic hydrops in all patients with this complication. CONCLUSIONS: Our data suggest that endolymphatic hydrops is a risk factor for recurrence of BPPV and that prevention of recurrent BPPV is important for control of endolymphatic hydrops. PMID- 19128713 TI - High-dose ropivacaine versus bupivacaine for posttonsillectomy pain relief in adults. AB - OBJECTIVE: To compare the efficacy and safety of preincisional high-dose ropivacaine with bupivacaine in relieving posttonsillectomy pain. DESIGN: A prospective, randomized, double blind, placebo-controlled clinical trial. SETTING: University hospital. PATIENTS: Before the incision, 58 subjects were randomly assigned to receive 10 mL of 10 mg/mL ropivacaine hydrochloride with epinephrine (n=19, group 1), 10 mL of 2.5 mg/mL bupivacaine with epinephrine (n=20, group 2), or saline with epinephrine (n=19, group 3) as a placebo. MAIN OUTCOMES MEASURES: Postoperative pain, additional analgesic drug consumption, otalgia, operating time, amount of intraoperative blood loss, and possible complications were assessed. The intensity of pain was scored on a visual analogue scale. The patients were followed up for 10 days after surgery. RESULTS: There were no statistically significant differences between the demographic and operational characteristics of the patient groups. Statistically significant differences were determined between in the mean intensity of constant and swallowing pain values of group 1 and group 2 or 3, at 4, 8, and 12 hours, and on days 1, 2, 3, and 4, postoperatively. However, no statistical differences were determined between the groups on days 5, 7, and 10. A lower amount of additional analgesic drug was consumed by the ropivacaine group than by the bupivacaine group, and by the bupivacaine group than by the control group, and the differences were statistically significant. CONCLUSION: Preincisional infiltration of the tonsils with high-dose ropivacaine markedly decreased the intensity of pain after tonsillectomy when compared with bupivacaine or placebo, especially until postoperative day 4 in adults. PMID- 19128714 TI - Assessment of long- and short-term neurotoxic effects of glass ionomer bone cement by electromyography and histopathologic examination: experimental study. AB - OBJECTIVE: The objective of this study was to investigate the possible neurotoxic effects of bone cement on the peripheral nerves. STUDY DESIGN: Experimental study. SETTING: Teaching and research hospital. METHODS: Twenty New Zealand rabbits were included in this study. The sciatic nerves of both legs of the 10 rabbits were exposed surgically under general anesthesia and closed primarily without any intervention and constituted the control group (group 1). Following surgical exploration, glass ionomer cement (GIC) was applied to the left sciatic nerves of the 10 rabbits for 10 seconds and then aspirated (group 2). GIC material was also applied to the right sciatic nerves of these rabbits but without aspiration (group 3). OUTCOME MEASURES: All rabbits were sacrificed at the end of 8 weeks postoperatively following electromyographic investigation. The sections were stained with hematoxylin-eosin and Immune Olig 2 staining technique for histopathologic examination under light microscopy. RESULTS: There was no statistically significant difference in distal latency, which indicates the conduction speed of the nerve, between all groups by electromyography. Histopathologic examination of all specimens revealed no demyelinization or axonal degeneration, and all had an intact myelin structure. There was no statistically significant difference in inflammation of the specimens between groups. (p>.05). CONCLUSION: GIC has no neurotoxic effects on the nerves in short and long-term applications. PMID- 19128715 TI - Pain and safety in otorhinolaryngologic procedures under local anesthesia. AB - OBJECTIVES: The aim of this study was to determine safety, pain perception, and the factors that may affect pain perception in patients undergoing certain otorhinolaryngologic procedures under local anesthesia with sedation. DESIGN: Prospective clinical study. SETTING: Tertiary care academic medical centre. METHODS: Two hundred thirteen patients operated on under local anesthesia with sedation were included in the study. A visual analogue scale (VAS) and a questionnaire were employed to measure pain perception during surgery and evaluate possible pain-related variables. RESULTS: The mean VAS values were 2.86 (+/-1.84) for the myringoplasty group, 2.54 (+/-1.76) for the otosclerosis surgery group, 1.70 (+/-1.25) for the somnoplasty group, 1.00 (+/-0.94) for the oral biopsy group, 2.15 (+/-1.49) for the inferior turbinate radiofrequency reduction (ITRR) group, 3.47 (+/-2.18) for the septoplasty plus ITRR group, 5.56 (+/-2.01) for the septorhinoplasty group, 4.94 (+/-2.16) for the open technique septoplasty group, and 3.30 (+/-1.87) for the septoplasty group. No variable was shown to affect pain perception when all patients were evaluated in common. When surgical procedures were evaluated individually, the level of acquaintance of patients in the myringoplasty group (p=.007) and operative time for the septoplasty group (p<.001) were found to independently affect pain perception. CONCLUSIONS: Given that septorhinoplasty and open technique septoplasty are found to be more painful, it is advisable to perform these procedures under general anesthesia. To increase the level of acquaintance of patients and to shorten operative time as possible may be helpful for minimizing pain perception. PMID- 19128716 TI - Evaluation of the effects of ethanol on static and dynamic gait. AB - OBJECTIVE: We used two balance assessment devices, computerized dynamic posturography (CDP) and Swaystar transducers to detect subtle balance system deficits in nine subjects who had ingested minimal amounts of alcohol. DESIGN: Nine subjects were evaluated with both modalities before, and repetitively after, ingesting a small amount of alcohol. METHODS: We measured condition 5 (sway referenced platform; eyes closed) on CDP and tandem walking with eyes closed while wearing Swaystar to see if either test could detect a balance deficit. MAIN OUTCOME MEASURES: We measured total sway amplitude with eyes closed in pitch and roll planes during tandem walking with Swaystar, and static balance scores of CDP sensory organization testing condition 5 before and after alcohol ingestion at 20 min intervals. RESULTS: Although there was no detectable deficit measurable by CDP, eight of our nine subjects showed increased dynamic sway as measured by Swaystar, after alcohol ingestion. Total sway was significantly greater (p=.05) after alcohol ingestion. CONCLUSION: It is important to assess dynamic, rather than static, equilibrium as it may have potential in detecting very subtle balance deficits. PMID- 19128717 TI - Increasing the usefulness of tandem walking evaluation. AB - OBJECTIVE: Tandem walking testing is a "standard" clinical technique for assessing balance and gait, but it is not a standardized test. We wished to develop a protocol by which we could measure sway during tandem walking at a patient's preferred cadence and at an altered cadence, which might be more challenging. DESIGN: Fifteen patients with vestibular complaints were evaluated with standard tandem walking testing while wearing the Swaystar belt-mounted accelerometer and were also assessed in the same way with tandem walking at two altered cadences. METHODS: We measured tandem walking sway amplitude and sway velocity with eyes open in both pitch and roll planes at a patient's preferred cadence, at 75% of their preferred cadence, and at 125% of their preferred cadence. MAIN OUTCOME MEASURES: We measured total sway amplitude and sway velocity in pitch and roll planes during tandem walking with eyes open while wearing Swaystar to see if there was any increase in sway at nonpreferred cadences. RESULTS: There was no correlation between preferred cadence and the age of the patient. However, there was a significant increase in both sway amplitude and sway velocity at both of the nonpreferred cadences. CONCLUSION: Disruption of a patient's preferred cadence may present an unnatural task, and this challenge to a patient's innate gait may help detect subtle vestibular disease. PMID- 19128718 TI - Actinomycosis in the etiology of recurrent tonsillitis and obstructive tonsillar hypertrophy: answer from a histopathologic point of view. AB - OBJECTIVE: To investigate the histopathologic profile and clinical presentation of tonsillar disease in the presence of Actinomycetes in children. DESIGN: A qualitative and quantitative histopathologic analysis of the palatine tonsil was performed. SETTING: Tonsillectomy specimens from patients who underwent tonsillectomy or adenotonsillectomy were searched for Actinomycetes. METHODS: Histologic evaluation of the specimens was done on hematoxylin and eosin-stained slides as blinded to patients' clinical category for disease groups. MAIN OUTCOME MEASURES: Four histologic compartments of the tonsil, including the surface epithelium, reticulated crypt epithelium (lymphoepithelium), lymphoid tissue, and interfollicular region, were examined. RESULTS: Actinomycetes was more prevalent in patients with obstructive symptoms treated with adenotonsillectomy. The number of subjectively quantitated total lymphoid follicles and small and medium-sized lymphoid follicles of the palatine tonsil were significantly increased in patients demonstrating "sulphur granules" in their crypts. Highly thick squamous metaplasia of the lymphoepithelium and dilatation of crypts were more prevalent in tonsil tissue revealing Actinomycetes. CONCLUSIONS: The presence of Actinomycetes in tonsillectomy specimens does not indicate active tissue infection. However, the histopathologic outcome may indicate the possible etiologic role of Actinomycetes in the development of prominent lymphoid hyperplasia and hypertrophy and, in turn, obstructive tonsillar hypertrophy. PMID- 19128720 TI - Effect of testing position on dynamic visual acuity. AB - OBJECTIVE: To investigate the effect of altering testing position on dynamic visual acuity (DVA) in vestibulopathic adults. STUDY DESIGN: Quasiexperimental research. SETTING: Outpatient vestibular program. PATIENTS: Twelve vestibulopathic adults with impaired DVA in the sitting position. METHODS: DVA scores were collected during (1) sitting, (2) standing comfortably, and (3) semitandem standing as subjects viewed a vision (E) chart 3.48 m in front of them. DVA was obtained as the head was passively moved by the evaluator at a frequency of 1.5 Hz in both horizontal and vertical directions. A further subgroup of subjects (n=5) was retested after performing the "viewing x1" gaze stability home exercise four times daily (1 minute duration in the horizontal direction while sitting) for 2 months. MAIN OUTCOME MEASURE: Noncomputerized DVA score. RESULTS: The DVA score remained constant independent of testing position as the Friedman test did not reveal any significant differences in the DVA scores in the horizontal or vertical direction. Subjects who improved after performing the viewing x1 exercise in the horizontal direction in the sitting position tended to have the same improvement in DVA in all three positions. CONCLUSIONS: The results of this study show that DVA score is independent of testing position. This is clinically important since it shows that the results of DVA testing in one position are valid for other positions and that subjects can be trained to improve their DVA in a sitting posture while having carry-over effects to other, more challenging postures. PMID- 19128719 TI - Effectiveness of itraconazole in the management of refractory allergic fungal rhinosinusitis. AB - OBJECTIVES: Conventional management of allergic fungal rhinosinusitis (AFRS) after surgery consists of the use of steroids to immunomodulate the body's response to fungi. However, there are many side effects to prolonged steroid use, and some patients are unresponsive to standard treatment. The role of systemic antifungal drugs in AFRS is still largely unknown. This was a pilot study to evaluate the effectiveness of itraconazole, an oral antifungal drug, in the treatment of refractory AFRS. METHOD: Thirty-two patients with AFRS who had had surgery and were refractory to prednisone, steroid, and amphotericin B nasal sprays were treated with itraconazole for at least 3 months. They were evaluated with pre- and posttreatment endoscopic examinations, serum immunoglobulin E (IgE), and the 31-Item Rhinosinusitis Outcome Measure (RSOM-31) questionnaires. Monthly liver function tests were done to monitor for the hepatic side effects of itraconazole. RESULTS: Twelve cases had endoscopic improvement. Fifteen had no difference, and five had a worse endoscopic stage after 3 months. One patient had to stop treatment due to abnormal liver function tests. The mean pre- and posttreatment IgE levels were 581 microg/L and 766 microg/L, respectively. Subjectively, 9 patients (28%) reported a significant improvement, 9 (28%) had moderate improvements, and 14 (44%) reported little or no change. There was no correlation between the subjective and the endoscopic changes. CONCLUSION: Itraconazole may be useful as an adjunct in the management of AFRS. However, more studies, including a prospective randomized clinical trial, are required to determine if itraconazole is effective in the management of AFRS. PMID- 19128721 TI - Effects of topical mitomycin and trimetazidine on myringosclerosis. AB - INTRODUCTION: Myringosclerosis, one of the most common complications of ventilation tube placement, is a kind of tympanosclerosis and is defined as subepithelial hyalinization of the tympanic membrane. There are two arguments in the development of myringosclerosis: inflammation triggering the development of myringosclerosis and free oxygen radicals released during inflammation causing myringosclerosis. OBJECTIVE: The aim of the present study was to explore the effects on the development of myringosclerosis of mitomycin, which has anti inflammatory effects, and trimetazidine, which is believed to inhibit free oxygen radicals when given systemically. MATERIALS AND METHOD: The study was carried out on rabbits. Animals were divided into five groups, with six rabbits in each group: three control groups (paracentesis only, paracentesis+tube placement, and no intervention), a trimetazidine group, and a mitomycin group. Mitomycin (0.4 mg/mL) and trimetazidine (20 mg/mL) were applied topically to the tympanic membrane, and the presence and degree of sclerosis were graded histopathologically after Masson's trichrome staining. RESULTS: In the histopathologic examination, sclerosis that developed in the tympanic membranes of rabbits that had undergone paracentesis or paracentesis plus tube application or received trimetazidine was significantly more extensive than sclerosis in the membranes of unoperated animals and those that had been administered mitomycin. CONCLUSIONS: Paracentesis in rabbits, independent of tube placement, causes sclerosis of the tympanic membrane. Results show that topical use of mitomycin, due to its anti-inflammatory effect, had alleviating effects on myringosclerosis, whereas topical trimetazidine did not. PMID- 19128722 TI - Postoperative pain management with ketorolac in facial plastic surgery patients. AB - OBJECTIVE: To evaluate the benefit of intraoperative locally or intramuscularly injected ketorolac in combination with local anesthetic versus local anesthetic alone on postoperative pain in patients undergoing facial plastic surgery. METHODS: Prospective randomized trial of 140 consecutive patients undergoing facelift, and/or forehead lift at a private facial cosmetic surgery practice by one surgeon. Outcome measurement included the requirement for pain medication (none, morphine, acetaminophen with codeine, or plain acetaminophen) at 1, 12, and 24 hours postoperatively. RESULTS: Ninety-five patients receiving locally injected ketorolac required significantly less postoperative analgesic (p<.05) than both the 20 patients who received intramuscularly injected ketorolac and the 25 patients who received local anesthetic alone at 12 and 24 hours. Those patients receiving no ketorolac required significantly more (p<.05) analgesic 1 hour postoperatively. CONCLUSION: Intraoperative locally injected ketorolac is an effective and safe means of reducing postoperative analgesic requirements. PMID- 19128723 TI - Clinical parameters in obstructive sleep apnea: are there any correlations? AB - OBJECTIVE: To assess the correlation between clinical symptoms and examination findings with severity of obstructive sleep apnea (OSA). DESIGN: Prospective observational study. SETTING: Otorhinolaryngology sleep clinic in a tertiary referral centre. PARTICIPANTS: Consecutive patients referred to the sleep clinic with a suspected diagnosis of OSA. MAIN OUTCOMES MEASURES: Body mass index (BMI), Epworth Sleepiness Scale (ESS) score, tonsil size, modified Mallampati index (MMP), Muller manoeuvre, reflux finding score (RFS), apnea-hypopnea index (AHI), lowest oxygen saturation, and percentage of rapid eye movement sleep. RESULTS: Eighty patients (14 female, 66 male), with a mean age of 42.9 years (range 22 66), were included in the study. The mean BMI was 27.6 (range 20-39). A correlation was found between ESS score and anteroposterior pharyngeal wall collapse at the level of the soft palate during Muller manoeuvre (r=.3, p=.02). BMI was significantly associated with a history of hypertension (r=.3, p=.01), MMP (r=.3, p=.001), and lateral wall collapse at the level of the soft palate during Muller manoeuvre (r=.4, p=.0001). OSA severity correlated significantly with male gender (p=.02), history of hypertension (r=.5, p<.0001), BMI (r=.2, p=.03), MMP (r=.3, p=.003), and upper airway collapse during Muller manoeuvre. CONCLUSIONS: Clinical findings in patients with OSA do correlate with the severity of OSA and polysomnographic findings. The degree of upper airway collapse during Muller manoeuvre does correlate with AHI severity. Accuracy in predicting the presence and severity of OSA allows for greater cost-effectiveness in screening patients prior to polysomnography. PMID- 19128724 TI - Fabrication of a three-dimensional autogenous costal cartilage framework in auricular reconstruction: experience with 250 cases of Asian lobule-type microtia. AB - BACKGROUND: Ear reconstruction is the necessary choice for patients with congenital lobule microtia. The best results are still obtained by autogenous cartilage ear reconstruction. We developed a three-dimensional autogenous costal cartilage framework for auricular reconstruction with the purpose of achieving a new auricle of high fidelity. METHODS: The patient's sixth to eighth rib cartilages were harvested as auricular framework materials, which were divided into three laminates for carving fine anatomic structures. By connecting and fixing, a three-dimensional auricular framework with a concha, helix, antihelix, triangular fossa, and navicular fossa could be used for the auricular reconstruction. RESULTS: The results obtained were considered satisfactory with vivid individual appearances and neat stable structures. Six frameworks were exposed or modified due to hematoma, infection, or cutaneous necrosis. CONCLUSIONS: With a three-dimensional auricular framework, the minute anatomic structures cited above make the external ear silhouette more vivid. PMID- 19128725 TI - [Present and future of FARMACIA HOSPITALARIA]. PMID- 19128726 TI - [Research strategies of the Sociedad Espanola de Farmacia Hospitalaria]. PMID- 19128727 TI - [The impact of different renal function measuring methods on the dosages of meropenem, piperacillin/tazobactam and cefepime in critically ill patients]. AB - OBJECTIVE: Assesment of dosage deviations of three ss-lactam antibiotics eliminated through the kidneys (meropenem, piperacillin/tazobactam and cefepime) by comparison of two prediction formulae, Cockroft-Gault (CG) and Modification of Diet in Renal Disease (MDRD) with 24 h urinary creatinine clearance (CrCl(24h)), as a reference method. METHOD: 125 samples of 61 critically ill patients (each one with CG, MDRD y CrCl(24h) values) were classified in one of the five stages of the National Kidney Foundation (NKF) according to CrCl(24h). Dosage discrepancies for each antibiotic based on CG y MDRD were studied in reference to CrCl(24h) by percentage agreement and weighted kappa. At each of the NKF stages, daily dosage differences (Delta=DosisCG-DosisCrCl(24h); Delta=DosisMDRD DosisCrCl(24h)) and percentage of samples with dosage discrepancies by CG and MDRD in reference to CrCl(24h) were calculated. RESULTS: There were no statistically significant differences between the two prediction formulae in respect to CrCl(24h), achieving good degrees of concordance. Deviation percentages fluctuated between 15.2% and 28% and occurred mainly by underdosing on stages 1 and 2 and by overdosing on stages 4 and 5. CONCLUSIONS: The two renal function prediction formulae can be indistinctly used to optimize the ss-lactam antibiotics dose regimen, CG being the easiest one. PMID- 19128728 TI - [Modification of empirical antimicrobial regimen during the first 72 hours of hospitalisation]. AB - OBJECTIVE: The aims of this study were to determine the empirical antibiotic therapy used in patients admitted to the Emergency Department who were later hospitalised, and to describe the antibiotic changes during their first days of hospitalisation. METHOD: All 14-year-old patients admitted to the Emergency Department who were started on antibiotic therapy and subsequently were hospitalised for at least 72 hours in an in-patient hospital ward, were included in a prospective observational study. Patients underwent daily follow-up during the first three days of hospitalisation. The type of infection, microbiological data and empirical antibiotic therapy and its changes were registered. RESULTS: 225 patients were included in this study. The most frequent types of infection diagnosed were infection of the respiratory airways, pneumonia and skin and soft tissue infection. Amoxicillin-clavulanic acid was the most widely prescribed antibiotic followed by levofloxacin and third generation cephalosporins. Microbiological samples were taken in 80 (36%) patients. Of the 225 antimicrobial regimens started in the Emergency Department, 94 (42%) were changed during the first 72 hours of hospitalisation: 37 (16%) were completely modified, 31 (14%) were discontinued and antibiotics were added or stopped from the existing regimen in 26 cases (12%). Among these 94 patients whose treatment was changed, only in 40 (42%) there was a microbiological result for aiding in the adjustment of the antibiotic therapy. CONCLUSION: The frequency of early changes during inpatient hospitalisation to antimicrobial regimens which were initially prescribed in the Emergency Department is high. Microbiological results were rarely used to guide these changes. PMID- 19128729 TI - Pharmaceutical interventions in metabolic and nutritional follow-up of surgical patients receiving parenteral nutrition. AB - PURPOSE: To assess a control protocol concerning alterations in metabolic follow up parameters in the context of a pharmaceutical care program designed for surgical patients receiving parenteral nutrition, through determination of the impact of pharmaceutical interventions on associated metabolic complications. METHODS: Prospective interventional study of two-months' duration performed in surgical patients receiving parenteral nutrition. The study variables included predefined biochemical parameters within the metabolic-nutritional profile. Four categories were established to classify the degree to which each parameter was altered: a) no alteration (within normal range); b) alteration with no associated complication; c) moderate complication, and d) severe complication. The type of pharmaceutical intervention carried out included a direct intervention on their part or a recommendation. Statistical differences between the mean analytical values before and after the intervention were assessed by parametric and non parametric tests (P< .05). RESULTS: A total of 1055 analytical determinations corresponding to 44 patients were evaluated. Among them, 239 determinations (22.6%) presented some degree of alteration which corresponded to 162 complications. Complication is often defined whit more than one parameter. Ninety three (57.4%) corrective interventions were carried out by direct intervention and 16 (9.9%) by recommendation. The results showed statistically significant differences or significant trend when the purpose of the pharmaceutical direct intervention was to increase albumin, prealbumin, potassium or phosphate levels or to decrease C-reactive protein, glucose or triglycerides. Significant differences or significant trend were not seen when no intervention was performed. CONCLUSION: Despite the fact that the parameters assessed may have been influenced by factors other than the parenteral nutrition treatment received, our findings show that systematic monitoring of specific analytic parameters can be effective for attaining success in nutritional therapy, in terms of improvement in nutritional status and prevention and control of associated complications. PMID- 19128730 TI - [Coauthorship networks and institutional collaboration in Farmacia Hospitalaria]. AB - BACKGROUND: Scientific collaboration is necessary for the advance of science. The purpose of this study is to analyse collaboration between authors and Spanish institutions in scientific studies published in Farmacia Hospitalaria, applying methodology derived from the analysis of social networks. METHODOLOGY: The study identified pairs of authors and institutions co-authoring or co-signing the same works published in Farmacia Hospitalaria between 1998 and 2007, building collaboration networks using the TextToPajek and Networks-PAJEK programs. RESULTS: 448 articles were analysed, showing an average signature/ article index of 4.79. Applying a collaboration threshold of 3 articles, 26 clusters were formed with principal researchers being Jimenez Torres and Perez Ruixo (n=16 co authorships) and Ribas Sala and Codina Jane (n=15). Among the institutions, there was significant collaboration between the Complejo Hospitalario Virgen del Rocio and the Complejo Hospitalario Nuestra Senora del Valme (n=4), both in Seville, and between Hospitals Vall d'Hebron (Barcelona), the Hospital de Navarra, and the Universitat de Barcelona (n=4). DISCUSSION: Analysis of the collaboration networks in Farmacia Hospitalaria has made it possible to identify the groups of authors and institutions in the area, as well as their relationships in terms of research and scientific publications. We propose to analyse the changes in these groups over a period of time, as well as to identify collaboration patterns in other national and international journals. PMID- 19128731 TI - [Drugs treatment of hepatitis B]. AB - OBJECTIVE: To define the role of those drugs available for hepatitis B treatment and analyse current treatment guides prepared by the leading scientific societies in the field. METHODS: Bibliographic searches were carried out in the databases PubMed and EMBASE, using the search word <>, limited by <> plus <>, <> or <>, within the period 1991-2007. RESULTS: Six drugs are currently available: interferon alpha (conventional or pegylated), lamivudine, adefovir, entecavir and telbivudine. In normal practice, pegylated interferon has almost completely displaced the conventional variety. HBeAg+ patients with high ALT levels, low HBV DNA counts and genotypes A and B show the best response to interferon. Lamivudine achieves faster and more potent viral suppression than adefovir; its principal drawback is the resistance that some patients develop. Its role will probably decrease as entecavir and telbivudine become more widespread, as they are associated with less resistance. Adefovir is useful in decompensated patients and/or those resistant to lamivudine. Because of the response rates it obtains, entecavir could be the drug of choice for HBeAG+ patients, particularly those with higher viral loads. For HBeAg- cases, any drug can be used as a first-choice drug. The main difference between the treatment guides lies in the way they define the illness and the serum markers that indicate active replication: viral loads and HBeAG positivity. CONCLUSIONS: All of the drugs are capable of accomplishing short-term biochemical, viral and histological objectives. There is no unanimous opinion on which patients should be treated with which drugs, during what length of time, and what objectives are to be reached. PMID- 19128732 TI - [Atrioventricular block induced by high doses of risperidone]. PMID- 19128733 TI - [Hepatic cytolysis from infliximab]. PMID- 19128734 TI - [Use of antidepressants in patients with generalised anxiety disorder compared with other indications in primary care]. PMID- 19128735 TI - [Transurethral resection syndrome]. PMID- 19128736 TI - [Pharmacy-provided management of medicinal gases]. PMID- 19128738 TI - [Patent ductus arteriosus]. PMID- 19128739 TI - [Viral respiratory tract infections in the first six months of life]. AB - BACKGROUND: Respiratory syncytial virus and Influenza virus infections are known causes of hospital admission in infants. It is less well known the pattern of virus infections in infants under 6 months of age in the outpatient setting. OBJECTIVE: To describe the clinical and epidemiological pattern of community acquired viral respiratory infections in infants under 6 months. PATIENTS AND METHODS: A cohort of infants from the 8 and 9 Madrid Health Districts was followed by telephone calls every two weeks since birth during the epidemic winter season. Clinical and epidemiological data were collected in pre-designed questionnaires. Nasopharyngeal aspirate was obtained in every patient with symptoms compatible with respiratory infection. Diagnosis of the more common virus was made with direct immunofluorescence and nucleic acid amplification test (PCR). RESULTS: Were recruited 316 newborns. The 1,865 phone calls made (median 4 for every child), produced 106 visits, and the illness confirmed in 89 illness. Rhinitis (91%) and cough (69%) were the most common symptoms. Upper respiratory infection was the principal clinical diagnosis (84.5%), and 17 of the 72 samples (23.2%) were positive. Most common viruses were RSV (41.1%) and rhinovirus (35.2%). Of the children visited, 17 out of 106 (16%) (5.3% of the cohort) were admitted to hospital. Diagnoses were febrile syndrome and bronchiolitis. We did not find any epidemiological factor associated with viral respiratory infection in positive cases. CONCLUSIONS: In our population most of the respiratory infections in infants are minor and do not need hospital assistance. Rhinovirus and RSV are the major pathogens. We did not find any epidemiological factor associated with viral respiratory infection. PMID- 19128740 TI - [Lung function changes after haematopoietic stem cell transplantation]. AB - OBJECTIVE: To evaluate lung function abnormalities in children who underwent haematopoietic stem cell transplantation (HSCT) and to compare these abnormalities between autologous and allogenic transplantation. PATIENTS AND METHODS: Prospective observational study from 1996 to 2005. Ninety-three children receiving HSCT, 47 autologous and 46 allogenic, were included. Lung function tests were performed before transplantation and at 2, 6, 12 and 24 months afterwards. The following indices were determined: forced expiratory volume in 1 s (FEV1), FEV1/forced vital capacity (FVC), total lung capacity (TLC), and carbon monoxide diffusing capacity (DLCO). Paired Student's t-test was used for statistical analysis of data. RESULTS: Before HSCT, 6.8% of the children had FEV1<80%, 1% FEV1/FVC<80%, 7.8% TLC<80% and 13.5% DLCO<70%. At 2 months, FEV1/FVC, TLC and DLCO were significantly reduced, when compared to pre transplantation values (p=0.05, 0.011 and p<0.001, respectively). Lung function gradually improved from 6 months post-transplantation, but did not reach pre transplantation values at 24 months. No significant differences were found when comparing allogenic and autologous transplantation, apart from a lower FEV1/FVC value at 6 months (p=0.02) in the first group. CONCLUSIONS: An important proportion of children who undergo HSCT have early pulmonary abnormalities (at 2 and 6 months after transplantation) with partial recovery at 24 months. PMID- 19128741 TI - [Complete isosexual precocious puberty: clinical, pelvic ultrasound and laboratory features]. AB - OBJECTIVE: To determine whether initial presentation varies according to aetiology, whether such differences allow differential diagnosis between idiopathic and organic forms, and whether CNS imaging can be avoided in some patients with central precocious puberty (CPP). PATIENTS AND METHODS: Children referred for evaluation of precocious puberty were evaluated, and the subpopulation of children with CPP was enrolled in this prospective observational study. Clinical, laboratory and ultrasound features of 62 consecutive patients with CPP (5 boys and 57 girls) were recorded. We compared the characteristics of idiopathic (3 boys, 49 girls) and organic (2 boys, 8 girls) CPP. RESULTS: There were no differences in pubertal staging, age at puberty onset (7.0 [5.8-7.5] vs. 7.3 [5.1-8.3] years), bone age/chronological age ratio (1.26 [1.2-1.3] vs. 1.23 [1.1-1.3]), maternal menarche (11.7+/-0.2 vs. 11.7+/-0.6 years) between idiopathic and organic CPP, respectively. Organic CPP patients had a poorer height SD (0.35+/-0.4 vs. 1.6+/-0.1; p<0.01), predicted adult height, growth rate and growth rate SD (0.8+/-0.9 vs. 3.7+/-0.7). Girls with organic CPP had significantly higher oestradiol levels (47.5 [25-68] vs. 27 [14-43] pg/ml) than girls with idiopathic CPP. Pelvic ultrasound at the time of diagnosis revealed the presence of pubertal changes in internal genitalia in 43.9% of girls (37.2% idiopathic versus 62.5% organic CPP subpopulation; p=0.18). CONCLUSIONS: There is a clinical-ultrasound overlap between idiopathic and organic CPP. Imaging remains necessary in all cases of central precocious puberty, and ultrasound data should not be replaced by other diagnostic investigations. PMID- 19128742 TI - [Influence of maternal hypothyroxinaemia during pregnancy on psychomotor development]. AB - INTRODUCTION: The maternal thyroid function during early pregnancy plays a fundamental role in foetal brain development as synthesis of thyroid hormone does not begin until the 20th week of gestation. MATERIAL AND METHODS: Throughout the year 2002, 147 women in their 37th week of pregnancy were enrolled for the study. To evaluate their thyroid function, the serum concentrations of free T4 and of TSH were determined. After birth, the psychomotor development of their children was evaluated with the Mc-Carthy scales. RESULTS: The median value of free T4 was 9.37 pmol/l, being the data obtained from more than half of the sampled women below the hypothyroxinaemia threshold. Children born from mothers with T4 levels below percentile 10 showed a significantly lower score on the general cognitive index than those whose mothers had normal free T4 serum concentrations. A positive correlation was found between the values of maternal free T4 and the general cognitive index (r=0.43; p<0.01). CONCLUSIONS: The concentrations of maternal free T4 are important, not only during the first months of pregnancy, but all along the process to ensure adequate development of the foetal brain. PMID- 19128743 TI - [A study of variability in glycaemia in children and adolescents with diabetes mellitus type 1 on treatment with insulin glargine]. AB - OBJECTIVE: To determine the usefulness of insulin glargine (IG) to reduce hipoglycaemias and hyperglycaemic events in children and adolescents with type 1 diabetes. PATIENTS AND METHODS: In a retrospective/prospective study, 29 patients with a high number of non-severe hypoglycaemias, aged 3-18, and an average HbA1c of 8+/-0.7, received IG once daily plus regular insulin or rapid analogue before meals. Inclusion criteria were: a) previous treatment with NPH insulin; b) diagnosis of type 1 diabetes for at least 1 year before starting IG, and c) >3 blood glucose controls within a day. Incidence of severe and non-severe hypoglycaemic events, hyperglycaemic events, HbA1c values, body mass index, daily insulin dose before and after the institution of glargine therapy, were collected. Additionally, family were asked to complete a diabetes quality of life survey. RESULTS: 1,294+/-411 glycaemias/subject were obtained. Hypoglycaemic episodes were not reduced (5.9% vs 6.2%) and hyperglycaemic events remained unchanged. Fasting blood glucose levels decreased from 195.3+/-36.6 to 162.8+/ 25.8 in all patients (p<0.05) and a tendency a decrease in nocturnal hypoglycaemias was observed. The average HbA1c and total daily insulin doses also remained unchanged (0.8+/-0.2 UI/Kg/day). CONCLUSIONS: Using IG achieves a glycaemic control similar to NPH, with a tendency to decrease the frequency of nocturnal hypoglycaemias and an improvement in fasting glycaemia values. PMID- 19128744 TI - [Systematic review of the efficacy of racecadotril in the treatment of acute diarrhoea]. AB - OBJECTIVE: To estimate, through a systematic review of the literature, the efficacy of racecadotril in the treatment of acute diarrhoea. MATERIAL AND METHODS: Randomised trials carried out in children comparing racecadotril with placebo in terms of diarrhoea recovery, stools output and adverse effects were selected. Electronic databases (Medline, EMBASE, CENTRAL, CINAHL, mRCT, Pascal) and bibliographies of retrieved articles were searched, and the drug developer was contacted. Two authors independently assessed the quality of the retrieved articles and extracted the data. RESULTS: Two small sample size randomised trials (135 and 172 children) of moderate quality were selected. They included children with less than five days diarrhoea and aged between 3 months and 4 years. There was no difference in the proportion of children who recovered by day 5 (RR=0.73, CI 95% 0.29 to 1.81), although the stools volume during the first 48 hours was less in the racecadotril group (SMD=-0.65, CI 95% -0.88 to -0.52). There is no difference in the risk of vomiting (RR=1.16, CI 95% 0.64 to 2.12). CONCLUSION: The proportion of recoveries by the 5th day is the same, although the stool volumes during the first 48 hours are less in the racecadotril treated children. It would be interesting to study the efficacy in a primary care setting assessing the cure rate, the stool volumes and the admission rate to elucidate if there is room for this drug. PMID- 19128745 TI - [Macroamylasaemia in paediatrics]. AB - Macroamylasaemia should be considered in any patient with high plasma amylase, no clinical signs and negative additional investigations for pancreatic or parotid diseases. It is characterised by an increase in serum amylase due to circulating high molecular mass macrocomplexes, most often formed due the binding of the amylase to an immunoglobulin. With a normal renal function, a hyperamylasaemia without an increase in urine amylase suggests the diagnosis, and is confirmed by identifying the macromolecular components. It is an uncommon entity in paediatrics. It has been described as a casual finding associated to abdominal pain and to celiac disease. We report two paediatric cases of macroamylasaemia, and a review of the tests needed for its diagnosis. The better understanding of this biochemical anomaly allows us to differentiate it from other situations associated to hyperamylasaemia, in order to avoid additional invasive explorations and unnecessary treatments. PMID- 19128746 TI - [Evolution of imaging in renal vein thrombosis in the newborn]. AB - We present a newborn patient with renal vein thrombosis with a good clinical and radiological evolution, describing the pathological features obtained by several imaging tools. Doppler ultrasound is the imaging method of choice, even although magnetic resonance gives excellent diagnostic findings it should be reserved for those cases where the Doppler findings are inconclusive. PMID- 19128747 TI - [Genetic diagnostic criteria in cases of mental retardation and development of idiopathic origin]. AB - Different studies show that mental retardation affects 1-3% of the population, and in about 50 % of the cases the aetiology is unknown. The uncertainty on the aetiology, and recurrence, means that prevention of mental retardation can have serious, therapeutic, social, and even economic repercussions. The key is to obtain an accurate diagnosis, proving a clinical hypothesis by the accomplishment of the most suitable genetic tests. Due to the increasing development of the technology in the field of the genetics, and the availability of new tests, this article reviews the criteria established in the practice guidelines from different scientific societies (paediatric, neurological and genetic) with respect to their use in diagnosis and integrates them from the point of view of their use in mental retardation and developmental delay. PMID- 19128749 TI - [Spontaneous haematoma of the umbilical cord]. PMID- 19128748 TI - [First SIBEN clinical consensus: diagnostic and therapeutic approach to patent ductus arteriosus in premature newborns]. AB - OBJECTIVE: To report the process and results of the first neonatal clinical consensus of the Ibero-American region. DESIGN AND METHODS: Two recognized experts in the field (Clyman and Van Overmeire) and 45 neonatologists from 23 countries were invited for active participation and collaboration. We developed 46 questions of clinical-physiological relevance in all aspects of patent ductus arteriosus (PDA). Guidelines for consensus process, literature search and future preparation of educational material and authorship were developed, reviewed and agreed by all. Participants from different countries were distributed in groups, and assigned to interact and work together to answer 3-5 questions, reviewing all global literature and local factors. Answers and summaries were received, collated and reviewed by 2 coordinators and the 2 experts. Participants and experts met in Granada, Spain for 4.5 h (lectures by experts, presentations by groups, discussion, all literature available). RESULTS: 31 neonatologists from 16 countries agreed to participate. Presentations by each group and general discussion were used to develop a consensus regarding: general management, availability of drugs (indomethacin vs. ibuprofen), costs, indications for echo/surgery, etc. Many steps were learnt by all present in a collaborative forum. CONCLUSIONS: This first consensus group of Ibero-American neonatologists SIBEN led to active and collaborative participation of neonatologists of 16 countries, improved education of all participants and ended with consensus development on clinical approaches to PDA. Furthermore, it provides recommendations for clinical care reached by consensus. Additionally, it will serve as a useful foundation for future SIBEN Consensus on other topics and it could become valuable as a model to decrease disparity in care and improve outcomes in this and other regions. PMID- 19128750 TI - [Neck-tongue syndrome. Possible joint hypermobility as an aetiopathogenic process]. PMID- 19128751 TI - [Cerebral venous thrombosis in a chemotherapy patient with dysgerminoma]. PMID- 19128752 TI - [Left Morgagni hernia with hepatic content]. PMID- 19128753 TI - [Stridor in a renal transplant patient]. PMID- 19128754 TI - [Haemorrhagic shock syndrome and encephalopathy (I)]. PMID- 19128756 TI - [Valproic acid induced-idiosyncratic hepatotoxicity]. PMID- 19128755 TI - [Haemorrhagic shock syndrome and encephalopathy (II)]. PMID- 19128757 TI - [Listeria monocytogenes induced-chronic arthritis in an immunoincompetent child]. PMID- 19128758 TI - [Psychomotor delay in children adopted in Russia is not associated with lead]. PMID- 19128759 TI - [Aicardi-Goutieres syndrome. Review of the genetic bases and their mechanisms]. PMID- 19128760 TI - [Anales de Pediatria: new challenges, new structure]. PMID- 19128761 TI - [Improvement of intestinal function in cystic fibrosis patients using probiotics]. AB - INTRODUCTION: In some cases, cystic fibrosis may include intestinal inflammation and bacterial overgrowth. Probiotics are considered as immunomodulatory, anti inflammatory and microbiotic regulator substances. The aim of our study is to determine the prevalence of bacterial overgrowth in cystic fibrosis patients and try to improve the intestinal function with the administration of probiotics. PATIENTS AND METHOD: We examined 20 patients with cystic fibrosis (mean age 10.33, range 5 to 17 years). The expired hydrogen test with a 2 g/kg of 20% dextrose overload was performed on 10 patients. After the test, Lactobacillus rhamnosus LGG 10(11) CFU was administered twice daily for four weeks. Faecal near infrared spectroscopy (FENIR) of water, fat, nitrogen and sugar content in faeces was performed before and after probiotics administration. RESULTS: Five patients (50%) showed bacterial overgrowth. We obtained a positive correlation between the hydrogen test and steatorrhea (R = 0.57) and sugar in faeces (R = 0.52). The FENIR results pre-treatment vs post-treatment were: fat 6.2 g +/- 3.3 g vs. 4.9 g +/- 2.1 g (p < 0.05), sugar 6.7 +/- g 3.6 g vs. 5 g +/- 2.6 g (p < 0.05) and nitrogen 0.87 g +/- 0.27 g vs. 0.91 g +/- 0.14 g (NS) respectively. Thirteen patients (81.25%) had improved stool appearance and intestinal comfort and nine (56.25%) decreased the number of daily stools. CONCLUSIONS: Probiotics improved not only clinical but also biochemical intestinal function in cystic fibrosis patients. These could be given as a regular treatment in this type of patients and in those with bacterial overgrowth. PMID- 19128762 TI - [Efficacy of exclusive enteral feeding as primary therapy for paediatric Crohn's disease]. AB - INTRODUCTION: The primary nutritional therapy (PNT), which consists in the administration of exclusive enteral formula feeds (polymeric, semi-elemental or elemental formula) for a period of no less than 6-8 weeks, has proven to be effective in inducing clinical remission in children with Crohn's disease. The clinical remission does not always include histological remission or cure of the mucosa. Faecal calprotectin is closely correlated with endoscopic and histological findings but is slightly associated with clinical activity scores. PATIENTS AND METHODS: An observational prospective study including all patients under 14 years of age diagnosed with Crohn's disease between January 2002 and October 2007, and who were fed exclusively with polymeric formula (Modulen IBD, Nestle, Vevey, Switzerland) during the onset of the disease. Clinical controls were carried out (weight, height, body mass index [BMI) and the Paediatric Crohn's Disease Activity Index [PCDAI)) and faecal calprotectin was measured at the beginning and at weeks 4 and 8 of treatment. The clinical remission was defined as having a PCDAI less than or equal to 10. Faecal calprotectin values below 50 microg/g faeces were considered as normal. RESULTS: There were 14 patients (9 males), mean age at diagnosis of 10.74 +/- 2.56 years. At week 4, 71 % of patients (10/14) had achieved clinical remission and a decrease in faecal calprotectin levels that was not significant. After 8 weeks, 85 % of our patients were in clinical remission and faecal calprotectin values had declined significantly without reaching normal levels. CONCLUSION: Primary nutritional therapy administered over a period of 8 weeks is capable of inducing clinical remission and improving the degree of inflammation of the intestinal mucosa. PMID- 19128763 TI - [Family impact of rotavirus gastroenteritis in children under two years]. AB - INTRODUCTION: Acute gastroenteritis (AGE) in infants has a significant impact on the quality of life of their parents. MATERIAL AND METHODS: Cross-sectional study on the sociological family impact related to rotavirus AGE in children under 2 years. The study was carried out in 25 hospitals and 5 primary care centres in Spain. Sociodemographic, epidemiological and clinical data were recorded, as well as the symptomatology of AGE and its severity measured by the Clark scale. Stool samples were tested to determine rotavirus positive (RV+) or negative (RV-). The parents were asked to complete a a family impact questionnaire. RESULTS: Stool specimens were tested in 1087 AGE cases (584 RV+ vs 503 RV-). The 99.5 % of parents whose children were RV+ reported more worries vs. the 97.7 % of RV-, and RV+ had a higher importance score (p < 0.05). A higher percentage of RV+ parents and those with a high importance score reported more time dedicated to dehydration treatment (p < 0.05). The 82.5 % vs. 73.9 % had disruption of their household tasks, with more importance scores (p < 0.05). RV+ had a higher percentage and importance score than RV- ones in all aspects of their child's AGE symptoms, except loss of appetite. CONCLUSION: AGE produces important dysfunctional experiences in daily family life. According to parental perceptions, RV+ produces greater worries and dysfunctions in child behaviour. PMID- 19128764 TI - [Imaging studies in the first urinary infection with fever in infants: is voiding cystourethrography necessary?]. AB - INTRODUCTION: The presence of vesicoureteral reflux (VUR) in an infant with urinary tract infection (UTI), does not necessarily lead to an acquired renal injury. Only serious reflux can be a factor in fostering its appearance. OBJECTIVES: To assess whether in infants diagnosed for the first time with a UTI with a fever, a routine initial DMSA can be used as a screening method for detecting severe reflux and replace voiding cystourethrography (VCUG). PATIENTS AND METHODS: We retrospectively studied 162 infants under 2 years old admitted to our hospital due to having a ITU with fever for the first time (92 males and 70 females). In all cases, a renal ultrasound, DMSA and VCUG had been performed a few days after the diagnosis. RESULTS: Of the 162 patients, 62 (38 %) had VUR, of which 56 (90 %) were mild and 6 (10 %) were severe. The DMSA was abnormal in 26/100 patients without VUR (26 %), 12/56 with mild VUR (21 %) and 6/6 of those with severe VUR (100 %). DMSA sensitivity for detecting severe reflux was 100 % and specificity was 76 %. The positive predictive value and negative predictive value was 14 % and 100 % respectively. The positive likelihood ratio was 4.17 and the negative likelihood ratio was 0. CONCLUSIONS: It is unnecessary to carry out VCUG in an infant with an initial UTI, and a negative DMSA. PMID- 19128765 TI - [Temporal and bacteriological comparison of neonatal urinary infections]. AB - OBJECTIVE: To find out the epidemiological and management changes of urinary tract infection over the last years. PATIENTS AND METHODS: Descriptive retrospective review of patients with urinary tract infections (UTIs) admitted to our neonatal unit over an 11 year period. A temporal, microorganism group and voiding cystourethrography (VCUG) group comparison. RESULTS: We studied 106 cases (5 cases/1,000 alive newborns). The most commonly isolated microorganisms were Escherichia coli (81.1%), Enterococcus faecalis (6.6%) and Enterobacter cloacae (4.7%). Gentamicin had an overall sensitivity of 93.2%, followed by cefotaxime 91.3% and amoxicillin-clavulanic acid 89.5%. We did not found temporal differences in microorganisms or sensitivities. We found differences in C reactive protein, nitrate and leucocyte values in E. coli versus non-E. coli UTIs, as well as patterns of antibiotic sensitivity with more resistances by non E. coli microorganisms. No differences were found in patients with normal and pathological VCUG. We found ultrasound and VCGU disturbances in 35.2% and 21.4%, respectively. CONCLUSIONS: UTI is a common problem in our area. The most frequently isolated microorganism is E. coli. Non-E. coli UTIs have more resistances to usual therapies, although these did not show any increase in disease in the imaging tests. We found no increase in resistances throughout the study. PMID- 19128766 TI - [Anaemia and neutropenia in a cohort of non-infected children of HIV-positive mothers]. AB - INTRODUCTION: Mother-to-child HIV transmission is currently around 1% in western countries, due to prevention measures. Antiretroviral drugs do have adverse effects, anaemia and myelosupression caused by AZT being the most observed effects. In the present study, we analyse the prevalence of anaemia and neutropenia in an uninfected children cohort born to HIV-infected women. MATERIAL AND METHODS: We followed up 623 uninfected children belonging to the FIPSE cohort according to standardised protocols. This cohort groups 8 hospitals from Madrid and follows up HIV infected pregnant women and their children. Anaemia and neutropenia were defined according to the ACTG (AIDS Clinical Trails Group) toxicity tables. Children were classified according to prematurity, ethnic origin, birth weight, withdrawal syndrome, in-utero treatment and neonatal prophylaxis. Categorical variables were compared with the chi2 or the Fisher tests. RESULTS: Anaemia was observed in 188 (30.1%) children during follow-up and 161 (25.8%) had anaemia grade 2 or higher. Prematurity (p < 0.001), low birth weight (p = 0.005) and Highly Active Antiretroviral Treatment (HAART) with Protease Inhibitors (p = 0.016) were associated with higher percentages of anaemia in children. Nadir haemoglobin values were reached by 6 weeks of life and anaemia was transient and disappeared by six months of age. Neutropenia was present in 41.9% (261 children) and 22.7% of the children had moderate-severe neutropenia. Prematurity was again associated with neutropenia (p = 0.01) and low birth weigh was associated only with moderate-severe neutropenia (p = 0.023). African infants had a higher percentage of neutropenia than the rest of the children (50% vs. 44%), although the differences were not significant. The type of in-utero treatment did not appear to influence the neutropenia. Neutropenia was still present in 12.5% of infants at 18 months of age. The type of neonatal prophylaxis to prevent mother-to-child transmission (monotherapy, dual therapy or triple therapy) did not influence either cytopenia. CONCLUSION: In our series, the proportion of children with anaemia is high: 30.1% Prematurity, low birth weight and HAART with IP were associated with a higher proportion of anaemia, which was transient and had little clinical relevance. The proportion of children with neutropenia was higher (41.9%) and was associated with prematurity, low birth weight and African origin. The type of neonatal prophylaxis does not seem to influence the development of cytopenias. Persistence of neutropenia (without clinical significance) was observed in a small percentage of the children 12.5%, at 18 months of age. PMID- 19128767 TI - [Analgesia when taking heel-lance blood in the newborn]. AB - OBJECTIVE: To evaluate the pain in healthy newborns requiring blood test by a heel-prick procedure and compare different pain management methods. PATIENTS AND METHOD: We studied 150 term infants, in three randomised groups, from the Maternity Unit of our Hospital for a period of three months. The first group of 50 newborns, received no specific analgesic intervention during blood tests, except our usual nursing intervention ("facilitated tucking"). The second and third group (50 newborns), received non-nutritive sucking-placebo and non nutritive sucking-24% sucrose respectively. RESULTS: In the control group, the average score on the scale of discomfort was 3.92, moderate pain, causing a crying time of 51.72 seconds; the group receiving a non-nutritive sucking placeboscored 2.1, slight pain, 10.68 seconds crying, while the group receiving non-nutritive sucking-24 % sucrose, expressed a level of discomfort of 1.5 points, slight, with an average crying time of 10.70 seconds. The comparative results between the control group and groups of non-nutritive sucking on placebo and 24% sucrose, both showed significant differences in the scores of the scale of discomfort, as well as in the time crying (p < 0.001). The comparative analysis between groups of non-nutritive sucking sucrose and placebo showed no significant differences. CONCLUSIONS: The blood test by heel lance represents a painful procedure of moderate intensity capable of analgesic treatment. A proper nursing method, along with a complement of non-nutritive sucking during extraction, significantly decreases the discomfort and crying, it being unnecessary to consider other analgesics. PMID- 19128768 TI - [Retinopathy of prematurity screening in Spanish neonatal units]. AB - INTRODUCTION: A wide variety of retinopathy of prematurity (ROP) screening criteria exists in the different hospitals. In 1988, the benefits of cryotherapy treatment in severe ROP cases were demonstrated, and since then, some scientific societies have made recommendations on the screening and treatment of ROP. Within the Spanish scientific community, there are no specific recommendations on screening, diagnosis or treatment. OBJECTIVES: To describe the criteria used in the screening of retinopathy in premature newborns by those Spanish hospitals caring for babies with birth weights below 1,500 g. MATERIAL AND METHOD: A cross sectional study was performed based on a 50 question questionnaire on screening criteria and other related issues. This questionnaire was sent to those public and private neonatal units in Spain caring for babies with birth weights below 1,500 grams. The questionnaire was sent and returned between January 2002 and May 2003. RESULTS: Units in all of the 17 autonomous communities and both autonomous cities in territorial Spain (Ceuta and Melilla) responded. There was a 90% response rate. All the participating hospitals perform ROP screening. Only 62.9% of the hospitals follow a written protocol on screening. A large majority of hospitals (79.8%) perform a retinal eye examination on all newborns below 1500 g at birth. Half of the hospitals included in the study, 51.6%, determine the cut off point for performing the screening at 32 weeks of gestational age. Around 73% of the hospitals include the administering of oxygen supplements to premature babies as part of the criteria for screening, regardless of the babies' weight and gestational age. Weight, gestational age and oxygen are the only criteria used in 51% of the units. That all hospitals in Spain should establish some screening criteria was suggested by 90% of the responses. Only 10% use analgesia or sedation, other than topical, when examining the retina. CONCLUSIONS: Even though all the hospitals participating in the study screen for ROP, there is little agreement on which criteria should be followed when screening and on other aspects related to its practice. PMID- 19128769 TI - [Cutis marmorata telangiectatica congenita. Review of 33 cases]. AB - INTRODUCTION: Cutis marmorata telangiectatica congenita (CMTC) is an uncommon congenital vascular malformation characterized by the presence of a localized or generalized pattern, frequently asymmetrical, with reticulated, erythematous or violaceous macules, present at birth or shortly thereafter. PATIENTS AND METHOD: Retrospective study of 33 cases of CMTC diagnosed between 1994 and 2007 in our hospital. Clinical and follow-up data were recorded in all cases. In some patients additional tests were performed. RESULTS: Most of the patients (51.5%) were female. In 87.9% of the cases lesions were observed at birth. All the cases were sporadic. CMTC was localized in 72.7% of the patients, being distributed mainly over the lower limbs. Associated anomalies were noted in 60.6%, with predominance of skin alterations (14 cases). Extracutaneous anomalies were present in 11 patients, most commonly atrophy and hypertrophy in the involved limbs. In none of the cases were there ocular or neurological manifestations. The average follow-up time was 14 months. Improvement of the lesions was observed in 45%, and in one case there was complete resolution. CONCLUSIONS: CMTC is a malformative disease which is frequently associated with other abnormalities. An appropriate assessment and follow-up of these patients is advisable. Performing a thorough medical history, including full physical examination is necessary. If the head is affected, ocular and neurological examination should be performed. The prognosis is generally good, with a tendency to improvement or disappearance in most cases. PMID- 19128770 TI - [Maturity onset diabetes of the young (MODY)]. AB - Hyperglycemia is an increasing cause of consultation in Paediatrics. Diagnosis depends on the context. Under the name of diabetes numerous clinical entities are included. MODY (maturity onset diabetes of the young) is, after type 1 diabetes, the most frequent cause of diabetes in childhood. For its diagnosis, a high degree of suspicion is needed and we have to be aware of the family history. PMID- 19128771 TI - [Childhood-onset diabetes treated with sulphonylureas]. AB - MODY 3 type diabetes belongs to the group of monogenic diabetes and is caused by mutations in the gene for hepatocyte nuclear factor 1-alpha (HNF1-alpha). Although MODY 2 type diabetes is the most frequent form of MODY diabetes in childhood, type 3 is the most frequent in the general population. We report the case of a 12 year old child with basal and post-prandrial hyperglycaemia. No cardinal symptoms of type 1 diabetes mellitus were present. There are numerous cases of diabetes in his family. C-Peptide was 1.13 ng/ml and pancreatic autoimmunity markers were negative. HNF-1alpha gene mutation was found in the patient as well as in his father and sister. Treatment with glibenclamide was started at a dose of 2.5 mg/day in order to reduce the risk of microvascular disease, as this as high in MODY 3 type diabetes as in type 1 diabetes mellitus. Blood glucose returned to normal and glycosylated haemoglobin was maintained between 4.9 and 5.6 %. Side-effects were not observed except some mild hypoglycaemias. PMID- 19128772 TI - [Anogenital warts: a clinical, pathological and virological study]. AB - The objective of this study was to investigate the possible relationship between the presence of anogenital warts (AGW) in children and the sexual abuse as mode of transmission. Our series includes 8 patients with AGW who were treated in our hospital during the year 2007. A complete physical examination was carried out, including colposcopy or anoscopy, and samples were taken for histopathological examination and human papiloma virus (HPV) subtyping. We considered perinatal transmission as a possible route in two cases. Although sexual abuse was definitively confirmed in only one case, we observed some findings in four cases that led us to consider the possibility of sexual abuse. We did not consider the possibility of heteroinoculation or autoinoculation from common warts in any case. Our results have demonstrated the difficulty in assessing with certainty the source of HPV contamination in children with AGW. PMID- 19128773 TI - [Non-invasive neonatal ventilation protocol: care with recommending too low pressures]. PMID- 19128774 TI - [Herpes simplex virus type 1 and peptic ulcer]. PMID- 19128775 TI - [Central-European encephalitis or tick-transmitted encephalitis vaccine in child travellers (response)]. PMID- 19128776 TI - [Familial transmission of community acquired methicillin-resistant Staphylococcus aureus infection]. PMID- 19128777 TI - [Cyproheptadine-induced urinary obstruction]. PMID- 19128778 TI - [Self-assessment of compliance with a cranioencephalic trauma protocol in the emergency department in children over 1 year old]. PMID- 19128779 TI - [Polymalformation syndrome in a neonate: VACTERL association with an extra spleen]. PMID- 19128780 TI - [Primary Cushing's syndrome: controversial aspects]. PMID- 19128781 TI - [Lactic acidosis induced by inhaling high doses of salbutamol]. PMID- 19128782 TI - [Sydenham's chorea: a past still present]. PMID- 19128783 TI - [Chronic angioedema associated with Giardia lamblia]. PMID- 19128784 TI - [Toxic shock as an initial sign of acute lymphoblastic leukaemia]. PMID- 19128785 TI - [Anterior cervical hyperthricosis]. PMID- 19128786 TI - Glimpses of the epigenetic landscape. AB - In this issue of Cell Stem Cell, Cui et al. (2009) provide snapshots of histone modification maps for two different stages of human blood cell development. Revealing chromatin-state maps in model cell types will provide an important framework in which to consider epigenetic controls at molecular and systematic levels. PMID- 19128787 TI - One breath closer to making engineered tissues a clinical reality. AB - Reported recently in Lancet, Macchiarini and colleagues (2008) implanted a living tissue-engineered airway in a female patient. The restoration of the patient's quality of life testifies to this successful translation of benchtop to bedside studies and provides promise for the application of regenerative medicine strategies to other clinical disorders. PMID- 19128788 TI - Hematopoietic stem cell quiescence: yet another role for p53. AB - p53, sometimes referred to as the "guardian of the genome," helps regulate cell cycle arrest, DNA-damage repair, apoptosis, and senescence. Adding to this list, in this issue of Cell Stem Cell, Liu et al. (2009) show that p53 also plays a role in regulating hematopoietic stem cell quiescence. PMID- 19128789 TI - Exposing hidden dimensions of embryonic stem cell cycle control. AB - In a recent issue of Nature Genetics, Wang and colleagues (2008) describe a mechanism for how the mir-290 microRNA cluster regulates the cell cycle of murine embryonic stem cells. A focal point of this regulation is the cyclin-dependent kinase inhibitor p21(Cip1). PMID- 19128790 TI - Wnt-related molecules and signaling pathway equilibrium in hematopoiesis. AB - There is near consensus that Wnt family molecules establish important gradients within niches where hematopoietic stem cells (HSC) reside. We review recent papers suggesting that a delicate balance is required between competing Wnt ligands and corresponding signaling pathways to maintain HSC integrity. Some steps in the transitions from HSC to lymphoid progenitor seem to be partially reversible and under the influence of Wnts. In addition, it has been recently suggested that HSC can oscillate between dormant versus active or lineage-biased states. We speculate that Wnts control a reflux process that may sustain stem cell self-renewal and differentiation potential. PMID- 19128791 TI - p53 regulates hematopoietic stem cell quiescence. AB - The importance of the p53 protein in the cellular response to DNA damage is well known, but its function during steady-state hematopoiesis has not been established. We have defined a critical role of p53 in regulating hematopoietic stem cell quiescence, especially in promoting the enhanced quiescence seen in HSCs that lack the MEF/ELF4 transcription factor. Transcription profiling of HSCs isolated from wild-type and p53 null mice identified Gfi-1 and Necdin as p53 target genes, and using lentiviral vectors to upregulate or knockdown the expression of these genes, we show their importance in regulating HSC quiescence. Establishing the role of p53 (and its target genes) in controlling the cell-cycle entry of HSCs may lead to therapeutic strategies capable of eliminating quiescent cancer (stem) cells. PMID- 19128792 TI - Tissue damage-induced intestinal stem cell division in Drosophila. AB - Stem cell division is essential for tissue integrity during growth, aging, and pathogenic assaults. Adult gastrointestinal tract encounters numerous stimulations, and impaired tissue regeneration may lead to inflammatory diseases and cancer. Intestinal stem cells in adult Drosophila have recently been identified and shown to replenish the various cell types within the midgut. However, it is not known whether these intestinal stem cells can respond to environmental challenges. By feeding dextran sulfate sodium and bleomycin to flies and by expressing apoptotic proteins, we show that Drosophila intestinal stem cells can increase the rate of division in response to tissue damage. Moreover, if tissue damage results in epithelial cell loss, the newly formed enteroblasts can differentiate into mature epithelial cells. By using this newly established system of intestinal stem cell proliferation and tissue regeneration, we find that the insulin receptor signaling pathway is required for intestinal stem cell division. PMID- 19128793 TI - Differential mobilization of subsets of progenitor cells from the bone marrow. AB - G-CSF stimulates mobilization of hematopoietic progenitor cells (HPCs) from bone marrow by disrupting the CXCR4/SDF-1alpha retention axis. We show here that distinct factors and mechanisms regulate the mobilization of endothelial (EPCs) and stromal progenitor cells (SPCs). Pretreatment of mice with VEGF did not disrupt the CXCR4/SDF-1alpha chemokine axis but stimulated entry of HPCs into the cell cycle via VEGFR1, reducing their migratory capacity in vitro and suppressing their mobilization in vivo. In contrast, VEGF pretreatment enhanced EPC mobilization via VEGFR2 in response to CXCR4 antagonism. Furthermore, SPC mobilization was detected when the CXCR4 antagonist was administered to mice pretreated with VEGF, but not G-CSF. Thus, differential mobilization of progenitor cell subsets is dependent upon the cytokine milieu that regulates cell retention and proliferation. These findings may inform studies investigating mechanisms that regulate progenitor cell recruitment in disease and can be exploited to provide efficacious stem cell therapy for tissue regeneration. PMID- 19128794 TI - Transplantation of human embryonic stem cell-derived photoreceptors restores some visual function in Crx-deficient mice. AB - Some of the most common causes of blindness involve the degeneration of photoreceptors in the neural retina; photoreceptor replacement therapy might restore some vision in these individuals. Embryonic stem cells (ESCs) could, in principle, provide a source of photoreceptors to repair the retina. We have previously shown that retinal progenitors can be efficiently derived from human ESCs. We now show that retinal cells derived from human ESCs will migrate into mouse retinas following intraocular injection, settle into the appropriate layers, and express markers for differentiated cells, including both rod and cone photoreceptor cells. After transplantation of the cells into the subretinal space of adult Crx(-/-) mice (a model of Leber's Congenital Amaurosis), the hESC derived retinal cells differentiate into functional photoreceptors and restore light responses to the animals. These results demonstrate that hESCs can, in principle, be used for photoreceptor replacement therapies. PMID- 19128795 TI - Chromatin signatures in multipotent human hematopoietic stem cells indicate the fate of bivalent genes during differentiation. AB - Histone modifications have been implicated in stem cell maintenance and differentiation. We have analyzed genome-wide changes in gene expression and histone modifications during differentiation of multipotent human primary hematopoietic stem cells/progenitor cells (HSCs/HPCs) into erythrocyte precursors. Our data indicate that H3K4me1, H3K9me1, and H3K27me1 associate with enhancers of differentiation genes prior to their activation and correlate with basal expression, suggesting that these monomethylations are involved in the maintenance of activation potential required for differentiation. In addition, although the majority of genes associated with both H3K4me3 and H3K27me3 in HSCs/HPCs become silent and lose H3K4me3 after differentiation, those that lose H3K27me3 and become activated after differentiation are associated with increased levels of H2A.Z, H3K4me1, H3K9me1, H4K20me1, and RNA polymerase II in HSCs/HPCs. Thus, our data suggest that gene expression changes during differentiation are programmed by chromatin modifications present at the HSC/HPC stage and provide a resource for enhancer and promoter identification. PMID- 19128796 TI - Effect of capsaicin-evoked jaw-muscle pain on intramuscular blood-flow. AB - AIM: To investigate effect of capsaicin-evoked masseter-muscle pain on intramuscular blood-flow (BF) at rest and during contractions. METHODS: Eight healthy men (22-31 years) participated. BF was measured with Laser Doppler (Moor Instruments, UK) using a single-fibre probe inserted into the right masseter. Three BF probes were attached to the skin above right and left masseter and the right-middle finger. Subjects performed 30 s isometric contractions at 5%, 15%, and 25% of maximal voluntary contraction. After the contractions, capsaicin (0.1 mL, 100 microg/mL) was injected into the right masseter close to the fibre probe. When the pain sensation had disappeared, the series of submaximal contractions were repeated. BF data were sampled continuously, processed in 10s bins, and analyzed with repeated-measures ANOVAs. RESULTS: Intramuscular BF significantly increased immediately after capsaicin injection (P < 0.050) and rapidly (30 s) decreased to pre-injection values. A significant increase in cutaneous BF above the right masseter was observed (P < 0.050) and lasted for 10 min, while a significant BF decrease in the finger (P < 0.050) was observed. The contractions were associated with increases in intramuscular BF before and after the injection (P < 0.022) and the contraction levels were also associated with increase in intramuscular BF before injection (P=0.008) but not after injection (P = 0.314). CONCLUSIONS: This study demonstrated BF increased by muscle contraction but failed to show effects of contraction levels on BF in a muscle exposed to nociceptive stimuli. Neurogenic inflammation in muscles could possibly be mediated via antidromical effects and local release of vasoactive substances. The decreased BF in the finger could be due to involvement of central regulatory mechanisms. PMID- 19128797 TI - Structure of a polysaccharide from the lipopolysaccharide of Vibrio vulnificus CECT4602 containing 2-acetamido-2,3,6-trideoxy-3-[(S)- and (R)-3 hydroxybutanoylamino]-L-mannose. AB - A polysaccharide was isolated by GPC after mild acid treatment of the lipopolysaccharide of Vibrio vulnificus CECT4602 and found to contain L-Rha, D GlcpNAc and 2-acetamido-2,3,6-trideoxy-3-(3-hydroxybutanoylamino)-L-mannose (L RhaNAc3NHb). GLC analysis of the trifluoroacetylated (S)-2-octyl esters derived by full acid hydrolysis of the polysaccharide showed that approximately 80% of the 3-hydroxybutanoic acid has the S configuration and approximately 20% the R configuration. The following structure of the polysaccharide was established by (1)H and (13)C NMR spectroscopies, including 2D ROESY and (1)H/(13)C HMBC experiments: [carbohydrate sequence see in text]. PMID- 19128798 TI - Stereoselective entry into the D-GalNAc series starting from the D-Gal one: a new access to N-acetyl-D-galactosamine and derivatives thereof. AB - A new stereoselective preparation of N-aceyl-D-galactosamine (1b) starting from the known p-methoxyphenyl 3,4-O-isopropylidene-6-O-(1-methoxy-1-methylethyl)-beta D-galactopyranoside (10) is described using a simple strategy based on (a) epimerization at C-2 of 10 via oxidation-reduction to give the talo derivative 11, (b) amination with configurational inversion at C-2 of 11 via a S(N)2-type reaction on its 2-imidazylate, (c) anomeric deprotection of the p-methoxyphenyl beta-D-galactosamine glycoside 14, (d) complete deprotection. Applying the same protocol to 2,3:5,6:3',4'-tri-O-isopropylidene-6'-O-(1-methoxy-1-methylethyl) lactose dimethyl acetal (4), directly obtained through acetonation of lactose, the disaccharide beta-D-GalNAcp-(1-->4)-D-Glcp (1a) was obtained with complete stereoselectivity in good (40%) overall yield from lactose. PMID- 19128800 TI - Multiple trauma: an ongoing evolution of treatment modalities? PMID- 19128799 TI - Neural computation as a tool to differentiate perceptual from emotional processes: the case of anger superiority effect. AB - Research findings in social and cognitive psychology imply that it is easier to detect angry faces than happy faces in a crowd of neutral faces [Hansen, C. H., & Hansen, R. D. (1988). Finding the face in the crowd - An anger superiority effect. Journal of Personality and Social Psychology, 54(6), 917-924]. This phenomenon has been held to have evolved over phylogenetic development because it was adaptive to quickly and accurately detect a potential threat in the environment. However, across recent studies, a controversy has emerged about the underlying perceptual versus emotional factors responsible for this so-called anger superiority effect [Juth, P., Lundqvist, D., Karlsson, A., & Ohman, A. (2005). Looking for foes and friends: Perceptual and emotional factors when finding a face in the crowd. Emotion, 5(4), 379-395; Purcell, D. G., Stewart, A. L., & Skov, R. B. (1996). It takes a confounded face to pop out of a crowd. Perception, 25(9), 1091-1108]. To tease apart emotional and perceptual processes, we used neural network analyzes of human faces in two different simulations. Results show that a perceptual bias is probably acting against faster and more accurate identification of anger faces compared to happy faces at a purely perceptual level. We suggest that a parsimonious hypothesis related to the simple perceptual properties of the stimuli might explain these behavioral results without reference to evolutionary processes. We discuss the importance of statistical or connectionist analysis for empirical studies that seek to isolate perceptual from emotional factors, but also learned vs. innate factors in the processing of facial expression of emotion. PMID- 19128801 TI - Normal body temperature and the effects of age, sex, ambient temperature and body mass index on normal oral temperature: a prospective, comparative study. AB - BACKGROUND: Body temperature is an indicator of health status. However, thermoregulatory function is thought to decline with aging. OBJECTIVES: To determine normal body temperature and the effects of age, sex, ambient temperature (AT), and body mass index (BMI) on normal oral temperatures (OTs). DESIGN: A prospective four group comparative descriptive design was used to compare four cohorts: young adults in summer, older adults in summer, young adults in winter, and older adults in winter. METHODS: The OT of 519 community dwelling older adults ages 65-95 and 540 younger adults ages 20-64 was compared. The OT was taken with an electronic thermometer between 8 a.m. and 10 a.m. during summer and winter in 2007 in Taipei, Taiwan. RESULTS: There was no difference in mean OT between the <65 and > or =65 groups measured during winter. However, the mean OT of the <65 group was 0.11 degrees C lower than the > or =65 group measured in the summer. Subjects (> or =85 years) had a higher correlation coefficient (r=0.48) between OT and AT than those in the 65-74 year older group (r=0.31) and 75-84 year older group (r=0.23). Moreover this study found that the mean OT of older females was higher than that of older males in both winter and summer cohorts. Finally, multiple regression analysis results indicated AT and sex were predictors of OT while age and BMI were not a significant predictor of OT. These four factors together accounted for 9.4% of the variance in the overall sample (age 20-95), 12.8%, in those 65-95 and 28.2% for those, > or =85 years old. CONCLUSIONS: These findings help to clarify discrepancies in the literature. The OT of those over age 65 and those 20-64 was lower than the accepted 37 degrees C norm. However, "older is colder" does not apply to all older adults. Our findings indicate AT and sex rather than age alone account for temperature variation in older adults with normal baseline temperature. Further investigation is needed to identify potential risk factors of impaired thermoregulation in older adults. PMID- 19128802 TI - Effect of verapamil, trandolapril and their combination on vascular function and structure in essential hypertensive patients. AB - AIM: The aim of the present study is to evaluate the effect of treatment with verapamil, trandolapril and their combination on peripheral microcirculation vasoreactivity. METHODS: Twenty hypertensive patients were randomized to receive oral trandolapril (4 mg oid; TRA) or verapamil (240 mg oid; VER) for 6 months and then the combination of the two drugs for additional 6 months. At baseline, 6 months and 12 months, peripheral microcirculation vasoreactivity was evaluated by forearm blood flow technique (venous plethysmography), as vasodilation to an endothelium-dependent (acetylcholine) and an endothelium-independent stimulus (sodium nitroprusside, SNP); minimal forearm vascular resistances (MFVR) were also evaluated. RESULTS: Blood pressure decreased similarly and progressively in both groups throughout the study period. In VER, 6-month verapamil treatment significantly increased vasodilation to acetylcholine, but not to SNP. The superimposition of trandolapril increased the response to SNP, and less to acetylcholine. In TRA group, 6-month treatment with trandolapril improved the response to SNP, but not to acetylcholine. In this group, the superimposition of verapamil caused a significant improvement in the response to acetylcholine, but not to SNP. At the end of the study, MFVR were significantly reduced in both groups, but to a greater extent in TRA. CONCLUSION: The present study demonstrates that chronic treatment with verapamil ameliorates endothelial function in the forearm microcirculation of essential hypertensive patients, while trandolapril protects microcirculation from structural alterations. The combination of the two drugs is potentially a powerful tool to counteract hypertension-related microvascular dysfunction and damage. PMID- 19128803 TI - Genotyping of alpha-thalassemia deletions using multiplex polymerase chain reactions and gold nanoparticle-filled capillary electrophoresis. AB - A gold nanoparticle-filled capillary electrophoresis method combined with three multiplex polymerase chain reactions (PCRs) was established for simultaneous diagnosis of five common alpha-thalassemia deletions, including the -alpha(3.7) deletion, -alpha(4.2) deletion, Southeast Asian (--(SEA)), Filipino (--(FIL)) and Thai (--(THAI)) deletions. Gold nanoparticles (GNPs) were used as a pseudostationary phase to improve the resolution between DNA fragments in a low viscosity polymer. To achieve the best CE separation, several parameters were evaluated for optimizing the separation conditions, including the capillary coating, the concentrations of polymer sieving matrix, the sizes and concentrations of GNPs, the buffer concentrations, and the pH. The final CE method for separating a 200-base pair (bp) DNA ladder and alpha-thalassemia deletions used a DB-17 capillary, 0.6% poly(ethylene oxide) (PEO) prepared in a mixture of GNP(32nm) solution and glycine buffer (25mM, pH 9.0) (80:20, v/v) as the sieving matrix with 1microM YO-PRO-1 for fluorescence detection; the applied voltage was -10kV (detector at anode side) and the separation temperature was 25 degrees C. Under these optimal conditions, 15 DNA fragments with sizes ranging from 0.2kb to 3.0kb were resolved within 11.5min. The RSDs of migration times were less than 2.81%. A total of 21 patients with alpha-thalassemia deletions were analyzed using this method, and all results showed good agreement with those obtained by gel electrophoresis. PMID- 19128804 TI - Serial displacement chromatofocusing and its applications in multidimensional chromatography and gel electrophoresis: II. Experimental results. AB - Part I of this study investigated the theory and basic characteristics of "serial displacement chromatofocusing" (SDC). In Part II of this study, SDC is applied to two prototype applications which have potential uses in proteomics and related areas involving the analysis of complex analyte mixtures. In the first application, SDC was used as a prefractionation method prior to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) to separate a human prostate cancer cell lysate. It was observed that the resolution achieved in narrow-pI-range 2D PAGE was improved when using SDC prefractionation, so that SDC may be useful as a low-cost, high-speed, and highly scalable alternative to electrophoretic prefractionation methods for 2D-PAGE. The second application involves the use of SDC as the first dimension, and reversed-phase chromatography as the second dimension, to produce a novel, fully automated, two-dimensional high-performance liquid chromatography technique. The method was shown to have performance advantages over one-dimensional reversed-phase chromatography for peptide separations. PMID- 19128805 TI - Serial displacement chromatofocusing and its applications in multidimensional chromatography and gel electrophoresis: I. Theory and general considerations. AB - The technique of "serial displacement chromatofocusing" (SDC) is investigated both theoretically and experimentally with model mixtures of proteins and peptides. The method employs a multistep, retained pH gradient formed using adsorbed buffering species to produce a series of discrete effluent fractions. Each of these fractions may contain several displaced protein bands under conditions of sufficient mass overloading, so that several displacement trains of adjoined bands can be produced in a single chromatographic run. Numerical simulations and experimental results showed selective concentration effects for minor components in a fraction when the feed amount was sufficient large. A computer-aided design method was developed to facilitate the use of the method and was applied to both anion- and cation-exchange column packings. Good agreement was achieved between the designed pH gradients and experimental results. The characteristics of SDC were also explored in terms of its loading capacity, scalability, repeatability, recovery, and differentiation of proteins between their true and apparent isoelectric point values. PMID- 19128806 TI - Using sound to study bubble coalescence. AB - Frothers are surfactants used in flotation to aid generation of small bubbles, an effect attributed to coalescence prevention. Studying coalescence at the moment of bubble creation is a challenge because events occur over a time frame of milliseconds. This communication introduces a novel acoustic technique to study coalescence as bubbles are generated at a capillary. The sound signal was linked to bubble formation and coalescence events using high-speed cinematography. The technique has the resolution to detect events that occur within 1-2 ms. The results show that for common flotation frothers and n-alcohols (C(4)-C(8)) coalescence prevention is not simply related to surface activity. A total stress model is used to give a qualitative explanation to the action observed. Results for salt (sodium chloride) are included for comparison. PMID- 19128807 TI - Phase behavior of aqueous polyion-surfactant ion complex salts: A theoretical analysis. AB - A simple thermodynamic theory is presented for water swollen complex salts formed by ionic surfactant and oppositely charged polyions. The description takes into account, on approximate level, free energy contributions from attractive and repulsive polyion-mediated interactions between the micelles, the mixing of micelles, polyion chains and water, and the hydrophobic effect. Explicit expressions for the chemical potentials of water, polyion, and surfactant ion are derived and used to calculate phase diagrams at various degrees of polymerization and linear charge density of the polyion and for surfactants with 12 and 16 carbons in the tail group. In all calculations the aggregation number is optimized and the phase structure (disordered or fcc) is determined. The effect of varying the charge densities of spheres interacting with a cross-linked polyion network is also investigated. Results from theory are compared with experimental and Monte Carlo simulation data reported in the literature. PMID- 19128808 TI - Surface micellization of poly(2-oxazoline)s based copolymers containing a crystallizable block. AB - The micellization on surfaces of copoly(2-oxazoline) diblock copolymers consisting of a crystallizable poly(2-nonyl-2-oxazoline) (pNonOx) block linked to a poly(2-ethyl-2-oxazoline) (pEtOx) block is investigated. Those micelles are not pre-existing in the initial ethanol solution but are formed during the spin coating process by the evaporation of the solvent inducing the precipitation of the less soluble pNonOx block. The morphology and size of the surface micelles vary according to the pNonOx molar fraction in the copolymers. Reorganization of the micelles and evolution of the surface energies after a thermal annealing are also studied. PMID- 19128809 TI - Phase behavior of semifluorinated catanionic mixtures: head group dependence and spontaneous formation of vesicles. AB - Hexadecyltrimethylammonium bromide (C(16)TAB)-sodium perfluorooctanoate (C(8)FONa) and hexadecylpyridynium bromide (C(16)PyB)-C(8)FONa catanionic semifluorinated mixtures have been studied by conductivity, dynamic light scattering (DLS), cryo-transmission electron microscopy (cryo-TEM) and polarizing microscopy. The regular solution theory, applicable for a limited fluorinated molar ratio, does not predict long-range electrostatic interactions. The results are consistent with the fact that in the hydrogenated-rich region the interaction is attractive in both catanionic mixtures. The systems containing pyridinium headgroups were of the stronger interaction. A transition from micelles was found in both mixtures as a function of fluorinated molar ratio. Special attention was devoted to the effect of the head group in the system properties. The information related with the mean vesicle radius measured by DLS was compared with the vesicle size distribution as well as the elastic properties of the bilayer measured with cryo-TEM. PMID- 19128811 TI - Letter to the editor: Borja et al.'s (2008) "Overview of integrative tools and methods ... worldwide" omits key elements. PMID- 19128810 TI - Effects of selective serotonin reuptake and dual serotonergic-noradrenergic reuptake treatments on memory and mental processing speed in patients with major depressive disorder. AB - Patients with major depressive disorder (MDD) usually suffer from altered cognitive functions of episodic memory, working memory, mental processing speed and motor response. Diverse studies suggest that different antidepressant agents may improve cognitive functions in patients with MDD. The aim of this work is to study the effects of serotonergic reuptake inhibitors (SSRIs) and serotonergic noradrenergic reuptake inhibitors (SNRIs) treatments to improve the performance on memory tasks and mental processing speed in MDD. Seventy-three subjects meeting criteria for major depressive disorder were assessed with the Hamilton depression rating scale and a neuropsychological battery. The subjects were medicated with escitalopram (n=36) or duloxetine (n=37) for 24 weeks. At the end of the trial, the subjects were assessed again with the same neuropsychological battery used prior to the treatment. Both treatments improved importantly the episodic memory and to a lesser extent, working memory, mental processing speed and motor performance. Our results suggest that cognition is partially independent from improvement in clinical symptoms. Both groups achieved remission rates in the HAM-D-17 after 24 weeks of treatment, but SNRI was superior to SSRI at improving episodic and working memory. Our work indicates that the superiority of SNRI over the SSRI at episodic memory improvement is clinically relevant. PMID- 19128812 TI - Effects of mariculture on macrobenthic assemblages in a Western Mediterranean site. AB - The effects of solid organic wastes from a marine fish farm on sediment was tested using macrobenthic fauna as biological indicators. Impact on benthic fauna was evaluated in the vicinity of a fish farm in the Tyrrhenian Sea (Western Mediterranean) between July 2001 and October 2002. Changes in benthic community structure were investigated using multivariate, distributional and univariate analyses (diversity indices, AMBI and M-AMBI). The results showed sharp disturbance of assemblages under the cages and no effects in the area more than 25 m from the cages. Sediment alterations were related to an increase in farmed biomass and its wastes, as well as to low current speed that allowed accumulation of organic matter on the sea floor. It was possible to follow the ecological succession from slightly altered assemblages to heavily polluted ones in the very short period of a single fish fattening cycle (15 months). PMID- 19128813 TI - Caring for children with cerebral malaria: insights gleaned from 20 years on a research ward in Malawi. AB - Clinicians treating patients with severe malaria in endemic areas confront a variety of challenges inherent to resource-poor settings, but it is possible to provide excellent care. The basic requirements, in addition to a thorough clinical examination of the patient, include assessing parasitaemia; determining anaemia (via haematocrit or haemoglobin); estimating blood glucose and lactate concentrations; establishing and maintaining i.v. access; measuring oxygen saturation and providing supplemental oxygen when necessary; grouping, cross matching and transfusing blood. This paper provides practical information on determining the Blantyre Coma Score, collecting cerebrospinal fluid and measuring the opening pressure, and administering controlled volumes of i.v. fluids. Included is a narrative protocol describing the approach to patients with cerebral malaria used on the research ward at the Queen Elizabeth Central Hospital in Blantyre, Malawi. PMID- 19128814 TI - Validity of the Weil-Felix test in the diagnosis of acute rickettsial infections in Sri Lanka. AB - The diagnosis of rickettsial infections in developing countries is based on clinical features and a positive Weil-Felix test (WFT), as tests such as indirect immunofluorescent antibody (IFA) assays are not available for routine use. We estimated the sensitivity of the WFT in Sri Lanka using IFA testing as the gold standard. The WFT demonstrated low sensitivity (33%) in diagnosing acute rickettsial infections and low specificity, with a positive titre of 1:320 seen in 54% of healthy volunteers and 62% of non-rickettsial fever patients. Therefore, the use of the WFT should be discouraged in the diagnosis of acute rickettsial infections. PMID- 19128815 TI - Acoustic wave transmission through piezoelectric structured materials. AB - This paper deals with the transmission of acoustic waves through multilayered piezoelectric materials. It is modeled in an octet formalism via the hybrid matrix of the structure. The theoretical evolution with the angle and frequency of the transmission coefficients of ultrasonic plane waves propagating through a partially depoled PZT plate is compared to finite element calculations showing that both methods are in very good agreement. The model is then used to study a periodic stack of 0.65 PMN-0.35 PT/0.90 PMN-0.10 PT layers. The transmission spectra are interpreted in terms of a dispersive behavior of the critical angles of longitudinal and transverse waves, and band gap structures are analysed. Transmission measurements confirm the theoretical calculations and deliver an experimental validation of the model. PMID- 19128817 TI - Effects of agricultural conditions on the leaching behaviour of veterinary antibiotics in soils. AB - Antibiotics may be released to soils during the application of manure as fertiliser to land. The compounds may subsequently be transported to and contaminate groundwater and surface waters. This paper describes a series of lysimeter-based studies to explore the leaching behaviour of three veterinary antibiotics (sulfachloropyridazine, oxytetracycline and tylosin) under different conditions that could occur in the agricultural environment. The specific objectives were to: (1) explore the influence of slurry amendment and incorporation on leaching; (2) assess the effects of climate on leaching behaviour; and (3) evaluate the predictive capability of a leaching model used in the regulatory assessment of veterinary medicines. Sulfachloropyridazine was detected sporadically in leachate at concentrations up to 0.66 microg L(-1) under typical irrigation conditions and more frequently at concentrations up to 8.5 microg L(-1) under extreme irrigation conditions. Incorporation and timing of rainfall had no effect on leaching behaviour. Oxytetracycline and tylosin were not detected in any leachate samples. These differences in behaviour were explained by the sorption and persistence characteristics of the compounds. Comparison of the experimental measurements with simulations from the leaching model indicated that the model greatly underestimates the transport of antibiotics to groundwater which raises questions over the application of these models in the regulatory risk assessment process. PMID- 19128816 TI - Infection and maturation of monocyte-derived human dendritic cells by human respiratory syncytial virus, human metapneumovirus, and human parainfluenza virus type 3. AB - Human respiratory syncytial virus (HRSV), human metapneumovirus (HMPV), and human parainfluenza virus type 3 (HPIV3) are common, important respiratory pathogens, but HRSV has a substantially greater impact with regard to acute disease, long term effects on airway function, and frequency of re-infection. It has been reported to strongly interfere with the functioning of dendritic cells (DC). We compared HRSV to HMPV and HPIV3 with regard to their effects on human monocyte derived immature DC (IDC). Side-by-side analysis distinguished between common effects versus those specific to individual viruses. The use of GFP-expressing viruses yielded clear identification of robustly infected cells and provided the means to distinguish between direct effects of robust viral gene expression versus bystander effects. All three viruses infected inefficiently based on GFP expression, with considerable donor-to donor-variability. The GFP-negative cells exhibited low, abortive levels of viral RNA synthesis. The three viruses induced low-to-moderate levels of DC maturation and cytokine/chemokine responses, increasing slightly in the order HRSV, HMPV, and HPIV3. Infection at the individual cell level was relatively benign, such that in general GFP-positive cells were neither more nor less able to mature compared to GFP-negative bystanders, and cells were responsive to a secondary treatment with lipopolysaccharide, indicating that the ability to mature was not impaired. However, there was a single exception, namely that HPIV3 down-regulated CD38 expression at the RNA level. Maturation by these viruses was anti-apoptotic. Inefficient infection of IDC and sub-optimal maturation might result in reduced immune responses, but these effects would be common to all three viruses rather than specific to HRSV. PMID- 19128818 TI - Defluoridation of drinking water by combined electrocoagulation: effects of the molar ratio of alkalinity and fluoride to Al(III). AB - The defluoridation efficiency (epsilon(F)) of electrocoagulation (EC) is closely related to the pH level of the F(-)-containing solution. The pH level usually needs to be adjusted by adding acid in order to obtain the highest epsilon(F) for the F(-)-containing groundwater. The use of combined EC (CEC), which is the combination of chemical coagulation with EC, was proposed to remove fluoride from drinking water for the first time in this study. The optimal scheme for the design and operation of CEC were obtained through experiments on the treatment of F(-)-containing groundwater. It was found, with OH(-) being the only alkalinity of the raw water, that the highest efficiency would be obtained when the molar ratio of alkalinity and fluoride to Al(III) (gamma(Alkalinity+F)) was controlled at 3.0. However, when the raw water contained HCO(3)(-) alkalinity, a correction coefficient was needed to correct the concentration of HCO(3)(-) to obtain the optimal defluoridation condition of gamma(Alkalinity+F)=3.0 for CEC. The correction coefficient of HCO(3)(-) concentration was concluded as 0.60 from the experiment. For the practical F(-)-containing groundwater treatment, CEC can achieve similar epsilon(F) as an acid-adding EC process. The consumption of aluminum electrode was decreased in CEC. The energy consumption also declined greatly in CEC, which is less than one third of that in the acid-adding EC process. PMID- 19128819 TI - Gleason grading of prostatic adenocarcinoma with glomeruloid features on needle biopsy. AB - Glomerulations in prostatic adenocarcinoma are characterized by dilated glands containing intraluminal cribriform structures with a single point of attachment, resembling a renal glomerulus. On prostate biopsy, glomerulations are exclusively associated with carcinoma and not associated with benign mimickers. However, the Gleason grading of carcinoma with glomerulations on needle biopsy remains controversial. We prospectively collected 45 prostate needle biopsies containing carcinoma with glomeruloid features from our consult files for a 9-month period and examined the association between glomerulations and the presence of concurrent high-grade carcinoma. Glomerulations were overwhelmingly associated with high-grade cancer on the same core, composed of either Gleason pattern 4 (n = 36, 80% of cases) or Gleason pattern 5 (n = 2, 4% of cases). Only a minority of glomerulations were surrounded exclusively by pattern 3 cancer (n = 7, 16% of cases) on the same core. Most of the cases with surrounding pattern 4 cancer were scored as 3 + 4 = 7 (n = 24, 66%), whereas a smaller fraction were scored as 4 + 3 = 7 (n = 9, 26%), and only a minority were 4 + 4 = 8 (n = 3, 9%). In most cases, glomeruloid change was present on the same core as the highest Gleason score carcinoma of the case. None of the pattern 3 cases and only a minority of the pattern 4 cancers had higher Gleason score carcinoma on additional cores (n = 5, 14%). Glomeruloid structures are a rare but diagnostic feature of prostatic carcinoma on needle biopsy. Our data indicate that glomerulations are overwhelmingly associated with concurrent Gleason pattern 4 or higher-grade carcinoma. In several cases, transition could be seen among small glomerulations, large glomeruloid structures, and cribriform pattern 4 cancer. These data suggest that glomerulations represent an early stage of cribriform pattern 4 cancer and, until follow-up data are available, are best graded as Gleason pattern 4. PMID- 19128820 TI - Effect of freezing method and storage at -20 degrees C and -70 degrees C on prothrombin time, aPTT and plasma fibrinogen levels. AB - Clinical and epidemiological trials often involve central laboratory analyses of coagulation tests, including fibrinogen, which requires freezing of the plasma samples. Although rapid freezing by immersion of sample tubes in liquid nitrogen and storage at -70 degrees C is recommended, plasma samples are often transferred directly to the storage compartments, and stored at -20 degrees C. Prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen using a kinetic fibrinogen assay, PT-derived fibrinogen, and an immunoassay were measured in fresh plasma samples from 16 healthy blood donors. In addition, four sets of aliquots were prepared. Set A was transferred directly to a -20 degrees C storage compartment, set B was first snap-frozen in liquid nitrogen and then transferred to the -20 degrees C compartment. Set C was transferred directly to a -70 degrees C freezer, set D was first snap-frozen in liquid nitrogen and then stored at -70 degrees C. Aliquots were thawed after one, two, three and four months storage and laboratory assays repeated. PT and aPTT were strongly influenced by freezing and storage. In contrast, freezing had little effect on fibrinogen levels. Differences were below three percent for all variants. Changes were smaller for samples stored at -70 degrees C compared to -20 degrees C, and for snap-frozen compared to not snap-frozen samples. Frozen and thawed samples generated slightly higher fibrinogen levels compared to fresh samples. Prothrombin time and aPTT should be measured in fresh samples, since freezing has an inconstant and unpredictable effect on the results. In contrast, freezing and storage has little effect on results of fibrinogen assays. A limitation of the study is that only samples from healthy blood donors were used. Plasma samples with abnormal fibrinogen concentration, or with abnormal concentrations of coagulation factors might behave differently. PMID- 19128821 TI - Human peripheral blood endothelial progenitor cells synthesize and express functionally active tissue factor. AB - INTRODUCTION: Endothelial progenitor cells are circulating cells able to home to sites of vascular damage and to contribute to the revascularization of ischemic areas. We evaluated whether endothelial progenitor cells synthesize tissue factor, a procoagulant protein also involved in angiogenesis. MATERIALS AND METHODS: Endothelial progenitor cells were obtained from the peripheral blood mononuclear fraction of normal donors and cultured in endothelial medium supplemented with specific growth factors. The procoagulant activity expressed by cells disrupted by freeze-thaw cycles was assessed by a one stage clotting assay. Tissue factor mRNA expression was evaluated by RT-PCR. RESULTS: Endothelial progenitor cells do not express procoagulant activity in baseline conditions. However, lipopolysaccharide induces the expression of procoagulant activity. The effect is dose-dependent and reaches statistical significance at 100 ng/mL lipopolysaccharide. Inhibition with an anti-tissue factor antibody and amplification of cDNA with primers based on the tissue factor sequence confirm the identity of this activity with tissue factor. The kinetics of tissue factor expression by endothelial progenitor cells is identical to that of human umbilical vein endothelial cells showing maximal activity within 4 hours, and then decreasing; in contrast, tissue factor expression by mononuclear cells lasts for longer times. Both 5,6-dichloro-beta D-ribofuranosyl-benzimidazole and cycloheximide prevented the expression of procoagulant activity. Stimulation of endothelial progenitor cells with tumor necrosis factor-alpha did not elicit any detectable procoagulant activity. CONCLUSIONS: Endothelial progenitor cells can be stimulated by lipopolysaccharide to synthesize tissue factor. This protein might be involved in thrombotic phenomena and might contribute to endothelial progenitor cells related neovascularization. PMID- 19128822 TI - Acquired activated protein C resistance is associated with IgG antibodies to protein S in patients with systemic lupus erythematosus. AB - The objective of this study was to clarify the roles of anti-phospholipid antibodies (aPLs) in the pathogenesis of acquired activated protein C resistance (APC-R) in patients with systemic lupus erythematosus (SLE). We examined several aPLs levels (lupus anticoagulant, anti-cardiolipin antibodies, anti-beta2 glycoprotein I antibodies, anti-protein C antibodies, and anti-protein S antibodies), the APC-R test, and the factor V Leiden test in 85 SLE patients. Acquired APC-R, which was not found in any patient with the factor V Leiden mutation, was present in 26 (30.6%) of 85 patients, and confirmed that acquired APC-R was a significant risk factor for thromboembolic complications [odd ratio (OR), 3.36; 95% confidence interval (CI), 1.24-9.11]. Multivariate logistic analysis revealed that both LA and anti-PS strongly associated with the presence of APC-R, and that the correlation between anti-PS and APC-R was much stronger (OR, 46.7; 95%CI, 6.99-311) than that between LA and APC-R (OR, 11.3; 95%CI, 2.26 57.0). Furthermore, the mean value of APC sensitivity ratios was significantly lower in SLE patients with anti-PS (mean +/- SD, 1.68 +/- 0.37, p < 0.0001) than in those without anti-PS (2.23 +/- 0.40). These results suggest that acquired APC R is most strongly attributable to functional interference of the APC pathway by anti-PS, which contribute to risk of thromboembolic complications. PMID- 19128823 TI - Protective effect of tetraethyl pyrazine against focal cerebral ischemia/reperfusion injury in rats: therapeutic time window and its mechanism. AB - INTRODUCTION: Tetramethyl pyrazine has been considered an effective agent in treating neurons ischemia/reperfusion injury, but the mechanism of its therapeutic effect remains unclear. This study was to explore the therapeutic time window and mechanism of tetramethyl pyrazine on temporary focal cerebral ischemia/reperfusion injury. MATERIALS AND METHODS: Middle cerebral artery occlusion was conducted in male Sprague-Dawley rats and 20 mg/kg of tetramethyl pyrazine was intraperitoneally injected at different time points. At 72 h after reperfusion, all animals' neurologic deficit scores were evaluated. Cerebrums were removed and cerebral infarction volume was measured. The expression of thioredoxin and thioredoxin reductase mRNA was determined at 6 and 24 h after reperfusion. RESULTS: Cerebral infarction volume and neurological deficit scores were significantly decreased in the group with tetramethyl pyrazine treatment. The expression of thioredoxin-1/thioredoxin-2 and thioredoxin reductase 1/thioredoxin reductase-2 was significantly decreased in rats with ischemia/reperfusion injury, while it was increased by tetramethyl pyrazine administration. CONCLUSIONS: Treatment with tetramethyl pyrazine, within 4 h after reperfusion, protects the brain from ischemic reperfusion injury in rats. The neuroprotective mechanism of tetramethyl pyrazine treatment is, in part, mediated through the upregulation of thioredoxin transcription. PMID- 19128824 TI - Clopidogrel induces an acute hemostatic deficit and increases intra abdominal bleeding in rabbits. AB - PURPOSE: Clopidogrel, a potent antiplatelet drug, increases hemorrhagic adverse events when its use is continued up to five days before cardiac surgery but data are lacking in non-cardiac surgery. We sought to determine the dose of clopidogrel which has a maximal antiplatelet and hemorrhagic effect in a rabbit model of non-cardiac surgery. METHODS: Twenty-four rabbits were divided into three groups according to the dose of clopidogrel administered (5, 10 and 20 mg.kg(-1)). Baseline measurement of platelet aggregation induced with ADP, platelet reactivity index (PRI) of the VASP-phosphorylation assay and hematologic variables were obtained the day before the experiment. Two hours after clopidogrel administration, the same variables were measured, along with intra abdominal bleeding following standardized hepato-splenic lesions. RESULTS: Platelet aggregation was inhibited in a dose-dependent manner: 46%+/-16% with 5 mg.kg(-1) and 93%+/-7% with 20 mg.kg(-1) of clopidogrel. PRI was reduced by 61%+/ 25% with 5 mg.kg(-1) of clopidogrel and by 92%+/-11% and 94%+/-10% with 10 mg.kg( 1) and 20 mg.kg(-1) respectively (p=0.01). Percentage reduction of platelet aggregation was positively correlated with the percentage reduction of PRI (r=0.69; CI(95), 0.40 to 0.86). Bleeding from hepato-splenic lesions was more important in the 10 and 20 mg.kg(-1) groups compared to the 5 mg.kg(-1) group (p<0.05). CONCLUSION: Higher doses of clopidogrel are associated with a more profound inhibition of platelet aggregation and PRI and increased blood losses following standardized hepato-splenic lesions. We conclude that our animal model demonstrates clopidogrel's propensity to increase intra abdominal bleeding after standardized hepato-splenic lesions and may help develop blood sparing strategies for patients undergoing surgery while on clopidogrel. PMID- 19128825 TI - Nervous control of circulation--the role of gasotransmitters, NO, CO, and H2S. AB - The origins and actions of gaseous signaling molecules, nitric oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H(2)S) in the mammalian cardiovascular system have received considerable attention and it is evident that these three "gasotransmitters" perform a variety of homeostatic functions. The origins, actions and disposition of these gasotransmitters in the piscine vasculature are far from resolved. In most fish examined to date, NO or NO donors are generally in vitro and in vivo vasodilators acting via soluble guanylyl cyclase, although there is evidence for NO-mediated vasoconstriction. Injection of sodium nitroprusside into trout causes hypotension that is attributed to a reduction in systemic resistance. Unlike mammals, NO does not appear to have an endothelial origin in fish blood vessels as an endothelial NO synthase has not identified. However, neural NO synthase is prevalent in perivascular nerves and is the most likely source of NO for cardiovascular control in fish. CO is a vasodilator in lamprey and trout vessels, and it, like NO, appears to exert its action, at least in part, via guanylyl cyclase and potassium channel activation. Inhibition of CO production increases resting tone in trout vessels suggestive of tonic CO activity, but little else is known about the origin or control of CO in the fish vasculature. H(2)S is synthesized by fish vessels and its constrictory, dilatory, or even multi-phasic actions, are both species- and vessel-specific. A small component of H(2)S-mediated basal activity may be endothelial in origin, but to a large extent H(2)S affects vascular smooth muscle directly and the mechanisms are unclear. H(2)S injected into the dorsal aorta of unanesthetized trout often produces oscillations in arterial blood pressure suggestive of H(2)S activity in the central nervous system as well as peripheral vasculature. Collectively, these studies hint at significant involvement of the gasotransmitters in piscine cardiovascular function and hopefully provide a variety of avenues for future research. PMID- 19128826 TI - Microtensile bond strength of resin-based composites to Ti-6Al-4V. AB - OBJECTIVE: The purpose of this study was to determine the microtensile bond strength of various resin composite/adhesive systems to alumina particle abraded Ti-6Al-4V substrate after aging for 24h, 10 days, and 30 days in distilled water at 37 degrees C. METHODS: Four laboratory resin composite veneering systems (Gradia, GR; Solidex, SOL; Ceramage, CER; and Sinfony, SF) were bonded to 25 mm diameter machined disks of Ti-6Al-4V with their respective adhesive and methodology, according to the manufacturer's instructions. Microtensile bars of approximate dimensions 1 mm x 1 mm x 6 mm were prepared for each resin composite/adhesive system. After cutting, groups (n=12) from each adhesive system were separated and either stored in water at 37 degrees C for 24h (baseline) or aged for 10 or 30 days prior to loading to failure under tension at a cross head speed of 1.0mm/min. Failure modes were determined by means of scanning electron microscopy (SEM). Statistical analysis was performed through one-way ANOVA and Tukey's test at 95% level of significance. RESULTS: Significant variation in microtensile bond strength was observed for the different systems and aging times. SOL and GR showed the highest mean bond strength values followed by SF and CER at baseline. Aging specimens in water had an adverse effect on bond strength for SOL and CER but not for the SF and GR groups. SIGNIFICANCE: In vitro bond strength of laboratory resin composites to Ti-6Al-4V suggests that strong bonds can be achieved and are stable for certain systems, making them useful as an alternative for esthetic fixed prosthetic restorations. PMID- 19128827 TI - Dental implants press fit phenomena: biomechanical analysis considering bone inelastic response. AB - OBJECTIVES: The paper presents an investigation of the biomechanical effects induced by oral implant insertion because of the press fit action caused by the difference between the major diameter of the implant and that of the drilled hole. The intent is to provide a correlation between bone-implant interaction and heavy stress-strain effects induced in bone tissues. In particular, the analysis allows for the evaluation of the bone mechanical response considering the inelastic range. MATERIALS AND METHODS: Bone material properties are defined, with regard to the elastic phase, according to a procedure that distributes, over the entire volume, the material constants magnitude and orthotropic directions deduced from experimental data. The inelastic phenomena are interpreted by a specific constitutive formulation and parameter identification procedure. The displacement field induced by implant insertion is provided by imposing the geometric configuration of the implant on the assumed drilled surface. RESULTS: Results are reported with regard to the displacement and strain fields caused in the peri-implant region. The range of strains induced in bone confirms that inelastic effects occur and their characterisation is essential for the interpretation of the press fit phenomenon. SIGNIFICANCE: The investigation allows for an accurate evaluation of the short term effects induced by press fit phenomenon in bone tissues which can be of particular interest also in consideration of immediate loading techniques in oral surgery. PMID- 19128828 TI - Thyroid lymphoma: beyond antigen stimulation. PMID- 19128829 TI - Unique cytogenetic features of primary myelodysplastic syndromes in Chinese patients. AB - Myelodysplastic syndromes (MDS) are a heterogeneous group of myeloid neoplasms. Chromosomal abnormalities have been detected in 40-70% patients with primary MDS and are heterogeneous among patients of different races and from different backgrounds. In the current study, 351 Chinese adult patients with primary MDS were retrospectively analyzed for their chromosomal abnormalities by karyotyping. Among the 237 cases (67.5%) of chromosomal abnormalities, 99 were copy number changes alone (41.7%), 70 were structural abnormalities alone (29.5%), and 68 displayed both of these changes (28.8%). Overall, the frequency of -5/5q /del(5)(q13-33) was 5.1% in these Chinese MDS patients, which was lower than that in the MDS patients of western countries (8.7-23.4%), and the incidence of 5q- syndrome was only 0.3% in Chinese MDS patients. On the other hand, the frequencies of trisomy 8 (19.1%) and -20/20q-/del(20)(q11-13) (9.4%) were higher than those in western countries (1.2-7.0% and 2.0-3.5%, respectively). Chromosomal translocations were also detected in 31 cases (13.1%) including 12 rare translocations that have not been reported in MDS patients before. In addition, i(17)(q10) was detected in nine cases (3.8%), of which six cases only had this single abnormality. According to the IPSS chromosomal prognostic classification, the incidence of poor-risk karyotypes increased in the advanced WHO subtypes (p < 0.001). Together, we detected the unique cytogenetic features of chromosomal abnormalities and some rare translocations of MDS among Chinese patients. PMID- 19128830 TI - Circulating blasts or myeloid precursor cells in peripheral blood can predict success of cytogenetic analysis. PMID- 19128831 TI - Y-chromosome loss as the sole karyotypic anomaly with 3'RARalpha submicroscopic deletion in a case of M3r subtype of acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by the presence of a chromosomal rearrangement involving retinoic acid receptor alpha (RARalpha) gene generating the X-RARalpha fusion. We describe here a unique RARalpha gene rearrangement in a patient with M3r subtype of APL. Conventional cytogenetic analysis revealed Y-chromosome loss as the sole karyotypic anomaly. No X-RARalpha fusion was detected by fluorescence in situ hybridization (FISH) using PML/RARalpha dual-color dual-fusion translocation probe set, or RARalpha dual color break apart rearrangement probe or reverse-transcription polymerase chain reaction (RT-PCR). However, FISH using RARalpha dual-color break apart rearrangement probe showed a deletion of the entire 3'-end of one allele of RARalpha gene. To our knowledge, this is the first documented APL with 3'RARalpha submicroscopic deletion which is not associated with X-RARalpha fusion. The molecular consequences of this anomaly remain to be elucidated. PMID- 19128832 TI - Polycyclic aromatic hydrocarbons and effects on marine organisms in the Istanbul Strait. AB - Surficial sediments and mussels (Mytilus galloprovincialis) from the Istanbul Strait and Marmara Sea were analysed for sixteen parent polycyclic aromatic hydrocarbon (PAH) contents by gas chromatography-mass spectrometry (GC-MS) employing isotope dilution technique. Microalgae toxicity testing was applied to sediment elutriates and biological responses in terms of filtration rate and lysosomal stability were measured in mussels. Total PAH concentrations ranged from 2.1 to 3152 ng g(-1) dry wt in sediments and from 43-601 ng g(-1) wet weight in mussels. Molecular indices of phenanthrene/ anthracene, fluoranthene/pyrene and benzo(a)anthracene/chrysene were used to differentiate between pyrolytic and petroleum origin. Results showed that most of the contamination originates from high temperature pyrolytic inputs with some slight contribution of petrogenic PAH. PAH in sediments were frequently lower than the National Oceanic and Atmospheric Administration (NOAA)-ERM (Effects Range Medium) index. Results of sediment elutriate toxicity testing and biomarkers indicate that the cause of negative effects in sediments may result from different classes of pollutants and does not only relate with PAH contamination. Mussels from most of the stations showed both reduced lysosomal membrane stability and filtration rate indicating disturbed health although the two biomarker results did not always complement each other. The effect studies showed that the pollutants in the strait ecosystem have more pronounced effects in the middle parts than those at the Black Sea entrance. PMID- 19128833 TI - L-type Ca2+ channels in mast cells: activation by membrane depolarization and distinct roles in regulating mediator release from store-operated Ca2+ channels. AB - Store-operated Ca(2+) channels (SOCs) are considered to be the principal route of Ca(2+) influx in non-excitable cells. We have previously shown that in mast cells IgE+antigen (Ag) induces a dihydropyridine (DHP)-sensitive Ca(2+) influx independently of Ca(2+) store depletion. Since the DHP receptor is the alpha subunit of L-type Ca(2+) channels (LTCCs), we examined the possible role of LTCCs in mast cell activation. Mast cells exhibited substantial expression of the alpha(1C) (Ca(V)1.2) subunit mRNA and protein on their cell surface. IgE+Ag induced Ca(2+) influx was substantially reduced by the LTCC inhibitor nifedipine, and enhanced by the LTCC activator (S)-BayK8644, whereas these agents had minimal effects on thapsigargin (TG)-induced Ca(2+) influx. These LTCC-modulating agents regulated IgE+Ag-induced cell activation but not TG-induced cell activation. Inhibition of SOCs by 2-aminoethoxydiphenyl borate reduced both degranulation and production of cytokines, including interleukin-13 and tumor necrosis factor alpha, whereas LTCC modulation reciprocally regulated degranulation and cytokine production. IgE+Ag, but not TG, induced substantial plasma membrane depolarization, which stimulated a DHP-sensitive Ca(2+) response. Moreover, IgE+Ag-, but not TG-induced mitochondrial Ca(2+) increase was regulated by LTCC modulators. Finally, gene silencing analyses using small interfering RNA revealed that the alpha(1C) (Ca(V)1.2) LTCC mediated the pharmacological effects of the LTCC-modulating agents. These results demonstrate that mast cells express LTCCs, which becomes activated by membrane depolarization to regulate cytosolic and mitochondrial Ca(2+), thereby controlling mast cell activation in a distinct manner from SOCs. PMID- 19128834 TI - Decrease in RelA phosphorylation by inhibiting protein kinase A induces cell death in NF-kappaB-expressing and drug-resistant tumor cells. AB - The RelA (p65) is a subunit of nuclear transcription factor kappa B (NF-kappaB) and actively participates in expression of NF-kappaB-dependent genes involved in inflammation and tumorigenesis. Hence, the regulation of p65 is an important strategy to regulate those responses. In this study, we provide data that the dichlorophenyl derivative of 1,2,4-thiadiazolidine (known as P(3)-25) induced cell death in NF-kappaB-expressing and doxorubicin-resistant cells. P(3)-25 inhibited NF-kappaB DNA binding activity partially, but inhibited NF-kappaB dependent genes expression completely. It inhibited phosphorylation of Rel A (p65) by inhibiting activity of protein kinase A (PKA). The PKA inhibition was independent of adenylate cyclase activity or cAMP level. The PKA activity decreased due to inhibition of catalytic subunit of PKA. P(3)-25 inhibited almost 80% PKA activity at 100 nM concentration, having an IC(50) at 10.5 nM. P(3)-25 potentiated different chemotherapeutic agents-mediated cell death. Our results suggest that P(3)-25 inhibits PKA activity followed by decreased phosphorylation of p65 and transcriptional activity of NF-kappaB thereby decreasing antiapoptotic proteins resulting in induction of apoptosis in NF-kappaB-expressing and doxorubicin-resistant cells. The study might help to understand the mechanism of P(3)-25-mediated apoptosis and to design it as new chemotherapeutic drug for tumor therapy. PMID- 19128835 TI - Global effects of inorganic arsenic on gene expression profile in human macrophages. AB - Inorganic arsenic, a major environmental contaminant, exerts immunosuppressive effects towards human cells. We previously demonstrated that relevant environmental concentrations of inorganic arsenic altered morphology and functions of human primary macrophages, suggesting interference with macrophage differentiation program. The goal of this study was to determine global effect of low concentrations of arsenic trioxide (As(2)O(3)) on gene expression profile in human primary macrophages, in order to identify molecular targets of inorganic arsenic, especially those relevant of macrophage differentiation process. Using a pan-genomic microarray, we demonstrate that exposure of human blood monocyte derived macrophages to 1microM As(2)O(3) for 72h, a non-cytototoxic concentration, results in up-regulation of 32 genes and repression of 91 genes. Among these genes, 26 are specifically related to differentiation program of human macrophages. Particularly, we validated that As(2)O(3) strongly alters expression of MMP9, MMP12, CCL22, SPON2 and CXCL2 genes, which contribute to major macrophagic functions. Most of these metalloid effects were reversed when As(2)O(3)-treated macrophages were next cultured in arsenic-free medium. We also show that As(2)O(3) similarly regulates expression of this macrophagic gene subset in human alveolar macrophages, the phenotype of which closely resembles that of blood monocyte-derived macrophage. In conclusion, our study demonstrates that environmentally relevant concentrations of As(2)O(3) impair expression of macrophage-specific genes, which fully supports interference of metalloid with differentiation program of human macrophages. PMID- 19128836 TI - Degradation-resistant protein domains limit host cell processing and immune detection of mycobacteria. AB - The Mycobacterium tuberculosis genome reveals a large family of glycine-alanine rich PE-PGRS proteins. Due to similarities with the glycine-alanine rich Epstein Barr nuclear antigen 1, there has been interest in whether PE-PGRS proteins inhibit cellular processing and presentation via the major histocompatibility complex class I pathway. We investigated whether PE-PGRS proteins were resistant to ubiquitin-proteasome-dependent degradation and CD8(+) T cell recognition. Upon transient expression of ubiquitin fusion constructs of either full-length Rv0978c(PE-PGRS) protein or its PE domain in HeLa cells, the former was markedly less susceptible to proteasomal degradation. When peptides of varying glycine and alanine content from different PE-PGRS proteins were fused to the N-terminus of SIINFEKL peptide, the alanine-rich fusions elicited lower interleukin-2 responses in SIINFEKL-specific CD8(+) T cells, with corresponding decrease in lysis of cells presenting such peptides. When CD8(+) T cells from Mycobacterium bovis BCG immunized mice were stimulated with either full-length PE-PGRS protein Rv3812 or its PE domain, the former exhibited a lower level of cytotoxicity against BCG infected autologous macrophages. These results suggest that mycobacterium PE-PGRS proteins have domains that confer resistance to ubiquitin-proteasome-dependent protein degradation, and the bacteria may have an abundance of such proteins to evade immune detection and killing of mycobacterium-infected cells. PMID- 19128837 TI - Genomic sequence encoding diversity segments of the pig TCR delta chain gene demonstrates productivity of highly diversified repertoire. AB - To better understand the function and diversity of gammadelta T cells, we determined the genomic sequence encoding diversity (D) segments of the porcine TCR delta chain and its upstream regions, because pigs and other artiodactyls have relatively high proportions of gammadelta T cells. The revealed sequence contained 28 variable (V) alpha/delta segments, including 4 TRDV1 and at least 6 Ddelta segments, a much higher number than in humans and mice. All 6 of the Ddelta segments that had canonical recombination signal sequences were functionally utilized in expressed TCR delta chain genes. The multiplicity of Ddelta segments enabled the use of more than 3 Ddelta segments in a single functional TCR delta chain. The increased number of TCR delta segments was acquired by the duplication of the germline sequence, which occurred after the divergence of artiodactyls from primates and rodents. These data demonstrate that the pig is able to generate a highly diversified repertoire of TCR delta chain molecules. PMID- 19128839 TI - Separation as a suicide risk factor. AB - BACKGROUND: Marital separation (as distinct from divorce) is rarely researched in the suicidological literature. Studies usually report on the statuses of 'separated' and 'divorced' as a combined category, possibly because demographic registries are not able to identify separation reliably. However, in most countries divorce only happens once the process of separation has settled which, in most cases, occurs a long time after the initial break-up. AIM: It has been hypothesised that separation might carry a far greater risk of suicide than divorce. The present study investigates the impact of separation on suicide risk by taking into account the effects of age and gender. METHODS: The incidence of suicide associated with marital status, age and gender was determined by comparing the Queensland Suicide Register (a large dataset of all suicides in Queensland from 1994 to 2004) with the QLD population through two different census datasets: the Registered Marital Status and the Social Marital Status. These two registries permit the isolation of the variable 'separated' with great reliability. RESULTS: During the examined period, 6062 persons died by suicide in QLD (an average of 551 cases per year), with males outnumbering females by four to one. For both males and females separation created a risk of suicide at least 4 times higher than any other marital status. The risk was particularly high for males aged 15 to 24 (RR 91.62). CONCLUSIONS: This study highlights a great variation in the incidence of suicide by marital status, age and gender, which suggests that these variables should not be studied in isolation. Furthermore, particularly in younger males, separation appears to be strongly associated with the risk of suicide. PMID- 19128838 TI - Inhibition of cancer cell growth by ruthenium(II) cyclopentadienyl derivative complexes with heteroaromatic ligands. AB - Inhibition of the growth of LoVo human colon adenocarcinoma and MiaPaCa pancreatic cancer cell lines by two new organometallic ruthenium(II) complexes of general formula [Ru(eta(5)-C(5)H(5))(PP) L][CF(3)SO(3)], where PP is 1,2 bis(diphenylphosphino)ethane and L is 1,3,5-triazine (Tzn) 1 or PP is 2x triphenylphosphine and L is pyridazine (Pyd) 2 has been investigated. Crystal structures of compounds 1 and 2 were determined by X-ray diffraction studies. Atomic force microscopy (AFM) images suggest different mechanisms of interaction with the plasmid pBR322 DNA; while the mode of binding of compound 1 could be intercalation between base pairs of DNA, compound 2 might be involved in a covalent bond formation with N from the purine base. PMID- 19128840 TI - Does sleep disturbance mediate neuropsychological functioning in older people with depression? AB - BACKGROUND: Depression in older adults is associated with neuropsychological dysfunction, fronto-subcortical brain changes and sleep disturbance. Research suggests that adequate sleep is critical for many aspects of cognition including processing speed, verbal skills and memory. However, the association between sleep disturbance and neuropsychological functioning in depression has not been well evaluated. The current study therefore aimed to investigate these relationships. METHODS: Forty-eight people (mean age=59.6, sd=8.2) meeting DSM-IV criteria for unipolar major depression were included for analysis. Neuropsychological assessment included assessment of processing speed, learning and memory, verbal fluency and executive functions. Early and late insomnia were defined by scores on the Hamilton Depression Rating Scale. RESULTS: While early insomnia was related to depression severity and poorer global cognition, late insomnia was associated with later age of depression onset, depression severity, and poorer scores on tests of verbal fluency and memory. The associations between cognition and late insomnia were not accounted for by depression severity or age of onset of disorder. LIMITATIONS: This study was retrospective in nature, and did not include objective measures of sleep. CONCLUSIONS: This is the first known study to indicate that late insomnia in older people with major depression may be independently and aetiologically linked to neuropsychological performance, particularly verbal fluency and memory. It may also indicate underlying structural and neurochemical changes. Sleep and circadian disturbance may serve as a biomarker for ongoing cognitive decline and may be a potentially modifiable risk factor. PMID- 19128841 TI - Modulation of ovine neutrophil function and apoptosis by standardized extracts of Echinacea angustifolia, Butea frondosa and Curcuma longa. AB - Impaired neutrophil function has been associated with increased infectious diseases in ruminants. Attachment of neutrophils to endothelium and superoxide production is critical features of their immune activity. Once the infection is cleared, programmed cell death ensures the rapid resolution of inflammation. To develop new natural therapeutics for ruminants, standard extracts of Echinacea angustifolia (Polinacea), Butea frondosa and Curcuma longa (Curcuvet) were first evaluated on ovine neutrophil functions. Curcuvet strongly reduced PMA-stimulated adhesion and superoxide production. Polinacea and B. frondosa extract also reduced these functions, but with less efficacy than Curcuvet. We analyzed the effect of extracts on spontaneous apoptosis and gene expression in neutrophils aged in vitro for up to 22h. IL8 is critical for neutrophil recruitment and the immune response; Bcl2-related proteins, Bcl2A1 and Bax, are key regulators of neutrophil fate. Spontaneous apoptosis strongly increased in ovine neutrophils cultured for 22h (T22), accompanied by an upregulation of IL8 and a decreased Bcl2A1:Bax ratio. Curcuvet stimulated spontaneous apoptosis and inhibited IL8 and Bcl2A1 gene expression at T22, whereas Polinacea and B. frondosa extract inhibited spontaneous apoptosis and stimulated IL8 expression at T22. These results suggest that Curcuvet has antiinflammatory activity, whereas Polinacea and B. frondosa have an immunomodulatory action on sheep neutrophils. PMID- 19128842 TI - Temporal aspects of laminar gene expression during the developmental stages of equine laminitis. AB - The results of recent studies indicate that inflammatory responses occurring in the early stages of equine laminitis lead to downstream events that eventually result in failure of the bond between the hoof wall and the distal phalanx. In order to gain further insights into the molecular mechanisms involved in the development of laminitis, an equine-specific cDNA microarray consisting of transcripts for more that 3000 genes was used to assess temporal changes in gene expression in laminar tissues at 1.5, 3 and 12 h after administration of either a laminitis-inducing agent (black walnut heartwood extract; BWHE) or an equal volume of water (control). As early as 1.5 h after BWHE administration, pro inflammatory genes associated with leukocyte activation and emigration, including MCP-3/CCL7, MCP-1/CCL2, IP-10/CXCL10 and ICAM-1 were up-regulated. At both 1.5 and 3h after administration of BWHE, expression of B-cell specific transcripts (e.g., Ig-gamma 3, Ig-gamma 1 and lambda-light chain) were decreased in the laminar tissues. At the onset of Obel grade 1 lameness in horses administered BWHE, other genes involved in inflammatory processes (e.g., serum amyloid A, calgranulin C and NFAT-activation molecule 1), regulation of inflammation (e.g., inter-alpha-trypsin inhibitor, BiP/GRP78 [Ig binding protein], L-plastin, serpin and nexin-1), antioxidant responses (e.g., superoxide dismutase), matrix turnover (e.g., MMP-9 and TIMP-1), and anti-microbial responses (e.g., serotransferrin, beta-defensin-1 and elafin) were up-regulated. These results provide convincing evidence that genes associated with inflammation, activation and extravasation of leukocytes, antimicrobial activities, and destruction of the lamellar basement membrane are induced during the early stages of development of laminitis in response to administration of BWHE. PMID- 19128843 TI - Effects of extremely low frequency electromagnetic fields on transient evoked otoacoustic emissions in rabbits. AB - OBJECTIVE: Humans are continuously exposed to an extremely low frequency (ELF) of electromagnetic fields (EMF), transmitted from the common sources like power stations, electric transmission lines, communication and radio-television signal transmission units. The present study aimed to assess the effects of 5.068 kV/m and 10.182 kV/m electric fields, which refer to the lower and upper intensity limits beyond which hazardous effects can be observed, on the auditory functions of rabbits via transient evoked otoacoustic emission (TEOAE) recordings. METHODS: The study was performed on 20 healthy adult female New Zealand White rabbits randomly divided into two groups and applied either 5.068 kV/m (Group 1) or 10.182 kV/m (Group 2) of electric field for 3h/day for 14 days. TEOAE responses were recorded on day 0 before the exposure (0-BE) and on the 6th (6th-AE) and 14th (14th-AE) days after the exposure (AE). Emission amplitudes at 1.0-4.0 kHz were analyzed. RESULTS: In Groups 1 and 2, the amplitudes separately recorded on the 6th-AE day were not different from the amplitudes recorded on day 0-BE. On the 6th-AE day, the only significant difference was detected in the right ear recordings of Group 1 at the frequency of 1.5 kHz (p=0.007). In Group 1, at 1.5 kHz, the median 6th day AE value (3.8 dB SPL) for the right ear was significantly lower than the median BE value. No significant difference in terms of amplitudes was detected in the comparison of 14th day AE with day 0-BE recordings. In the comparison of the groups for the recordings obtained at all the time points, no statistically significant differences were found. CONCLUSION: It was concluded that the TEOAE decrease at 1.5 kHz of the right ears of Group 1 on the 6th day AE was transient; and on the 14th day AE, no significant decrease was determined in the TEOAEs of both groups. Our results showed that the ELF EMFs have no significant effects on the hearing sensation of rabbits, the cochlear functions of whose were evaluated using TEOAE recordings. PMID- 19128844 TI - Optical coherence tomography after new scoring balloon angioplasty for in-stent restenosis and de novo coronary lesions. AB - The AngioSculpt scoring balloon catheter (AngioScore, Inc., Fremont, California) has recently been developed for percutaneous intervention in coronary and peripheral arteries. This device is composed of two major components, a minimally compliant balloon and three nitinol wore. The three wires encapsulate the low compliant balloon in a spiral configuration. The concept is for the spiral wires to score the lumen surface during balloon expansion. However, the precise mechanisms and efficacy of this scoring technology in humans had not yet to be determined. In this case, both a de novo coronary lesion and an in-stent restenosis lesion were treated with the scoring balloon and were subsequently observed via optical coherence tomography (OCT) with high-resolution images ( approximately 15 microm). OCT clearly demonstrated the effects of this device on plaque and neointimal hyperplasia scoring, as well as its ability to achieve sufficient lumen sizes after coronary artery dilatation. PMID- 19128845 TI - Primary cardiac tumors. AB - Primary cardiac tumors are a rare entity compared to tumors that metastasize to the heart. Patients with such tumors may be asymptomatic. Many cases are found incidentally during evaluation of an unrelated medical condition. It is important for the clinician to have a high index of suspicion when evaluating a patient presenting with signs and systemic symptoms concerning possible malignancy, plus cardiac specific symptoms or complications. These can include new onset dyspnea, congestive heart failure, arrhythmias or murmurs varying with body positions. Imaging, particularly the use of echocardiography, remains the cornerstone of diagnosis, and may be combined with new imaging modalities of cardiac CT and MRI. The aim of this paper is to describe the epidemiology and pathophysiology of the various benign and malignant primary cardiac tumors. PMID- 19128846 TI - Atrial fibrillation and amino-terminal pro-brain natriuretic peptide as independent predictors of prognosis in systolic heart failure. AB - BACKGROUND: Survival of patients with systolic heart failure (HF) may be influenced by the presence of chronic atrial fibrillation (AF) and circulating concentrations of B-type natriuretic peptides. In this study, we sought to assess the prognostic value of chronic AF in comparison to those of amino-terminal pro brain natriuretic peptide (NT-proBNP) plasma levels and of echocardiographic parameters among HF patients of the entire study population and in those with AF. METHODS: Plasma NT-proBNP levels and echocardiography were prospectively assessed in 489 patients with chronic systolic HF (LV ejection fraction or = 8% from 8.4% to 7.8% (p<0.05). CONCLUSIONS: In an unselected, but relatively well-controlled population of type 1 diabetes, a structured, but not very time consuming FIT teaching programme in the outpatient setting improves psychological well-being and metabolic parameters. PMID- 19128853 TI - Study of awareness among pregnant women of the effects of drugs on the fetus and mother in Iran. AB - OBJECTIVE: The aim of the study was to examine the awareness of Iranian pregnant women about the effects of drugs in pregnancy. METHODS: Awareness of 400 women in postnatal and prenatal wards was assessed using self-completion questionnaire, which included demographic information, medication use, and the level of information regarding the safety of drugs during pregnancy and the most susceptible periods in pregnancy. RESULTS: Out of 400 participants from prenatal and postnatal wards of two hospitals, 19.0% used medications other than vitamins and minerals supplements during pregnancy, and 7% believed in the safety of medications for mother and/or fetus during pregnancy. The first and second trimesters were believed to be the most and the least susceptible periods of pregnancy, respectively. Most information on drugs safety in pregnancy was obtained from physicians and health centers. CONCLUSIONS: Present study indicates weaknesses in the awareness of this population and weak role of pharmacists in informing this vulnerable population. This study shows the requirement of training of this group of people in order to enhance the health of our community. PMID- 19128854 TI - Intrathoracic hibernoma: report of two cases. AB - Hibernomas are uncommon benign soft tissue tumours mimicking brown fat. The most common anatomic locations include the neck, axilla, mediastinum, periaortic and perirenal zones. Intrathoracic and in particular pleural locations are exceptional. We report two cases of intrathoracic hibernoma with pleural involvement treated by surgical resection. PMID- 19128855 TI - Prognostic significance of visceral pleural invasion in the forthcoming (seventh) edition of TNM classification for lung cancer. AB - BACKGROUND: The next revision to the TNM classification for lung cancer (the seventh edition) is scheduled to be released in 2009. However, the definition of visceral pleural invasion (VPI), which is a non-size-based T2 descriptor, still lacks in detail, and its validation is not included. METHODS: We analyzed 1046 cases of non-small cell lung cancer (NSCLC) with T1, T2, or T3 diseases from 1990 to 2005, and subclassified into p0-p3 according to the degrees of pleural invasion. Survival analyses were performed using Kaplan-Meier method. Then, all patients were subdivided into nine groups according to tumor size and pleural invasion, and we compared survival differences, primarily focusing on T2a and T2b diseases according to the seventh edition. RESULTS: There was no survival difference between patients with p1 and p2, thus we regarded p1 or p2 as VPI. There was survival difference between two groups, which are expected to be classified as T2b. The behavior of tumors larger than 5cm but 7cm or less with VPI was similar to T3 tumors. CONCLUSIONS: VPI is a poor prognostic factor of NSCLC, and the penetration through the elastic layer of the visceral pleura regardless of its exposure on the pleural surface (pl and p2) should be defined as VPI. This study also indicates that VPI influences T stage dependent on tumor size, and it can be suggested that tumors of larger than 5cm but 7cm or less with VPI should be upgraded to T3 stage. PMID- 19128856 TI - Green tea, black tea consumption and risk of lung cancer: a meta-analysis. AB - Studies investigating the association of green tea and black tea consumption with lung cancer risk have reported inconsistent findings. To provide a quantitative assessment of this association, we conducted a meta-analysis on the topic. Studies were identified by a literature search in PubMed from 1966 to November 2008 and by searching the reference lists of relevant studies. Summary relative risk (RR) estimates and their corresponding 95% confidence intervals (CIs) were calculated based on random-effects model. Our meta-analysis included 22 studies provided data on consumption of green tea or black tea, or both related to lung cancer risk. For green tea, the summary RR indicated a borderline significant association between highest green tea consumption and reduced risk of lung cancer (RR=0.78, 95% CI=0.61-1.00). Furthermore, an increase in green tea consumption of two cups/day was associated with an 18% decreased risk of developing lung cancer (RR=0.82, 95% CI=0.71-0.96). For black tea, no statistically significant association was observe through the meta-analysis (highest versus non/lowest, RR=0.86, 95% CI=0.70-1.05; an increment of two cups/day, RR=0.82, 95% CI=0.65 1.03). In conclusion, our data suggest that high or an increase in consumption of green tea but not black tea may be related to the reduction of lung cancer risk. PMID- 19128857 TI - CGRP-alpha responsiveness of adult rat ventricular cardiomyocytes from normotensive and spontaneously hypertensive rats. AB - Calcitonin gene-related peptide (CGRP)-alpha is expressed in heart ventricles in sensory nerves and cardiomyocytes. It modifies inotropism and induces ischaemic preconditioning. This study investigates the effect of CGRP-alpha on the contractile responsiveness of isolated adult ventricular rat cardiomyocytes and the effect of chronic hypertension on this interaction. Cardiomyocytes were isolated and paced at 0.5-2.0 Hz. Cell shortening was recorded via a line camera with a reading frame of 500 Hz. CGRP-alpha exerted a dual effect on cardiomyocytes with a positive contractile effect at 10nM and a negative contractile effect at 10 pM. CGRP-alpha(8-37), a calcitonin receptor-like receptor (CRLR) antagonist, attenuated the positive contractile effect. H89, a protein kinase A antagonist, converted the positive contractile effect into a negative contractile effect. The negative contractile effect was converted again back to a positive contractile effect in the presence of l-nitro arginine. In cardiomyocytes isolated from spontaneously hypertensive rats (SHR) the mRNA expression of CRLR and the receptor-associated modifier protein (RAMP)-2 were lower. However, on the protein level CLRL was up-regulated, RAMP2 expression remained unchanged, and eNOS expression was down-regulated in these cells. These cells responded with a pure positive contractile response. In Langendorff preparations, CGRP-alpha slightly reduced the rate pressure product in hearts from normotensive rats but it caused an increase in hearts from SHR. In conclusion, it is shown that CGRP-alpha exerts dual effects on cardiomyocytes favouring the negative contractile effect at very low concentrations. This effect is compensated in chronic pressure-overloaded hearts and converted into a positive inotropism. PMID- 19128859 TI - 3D-QSAR studies on the inhibitors of AP-1 and NF-kappaB mediated transcriptional activation. AB - Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on a series of 68 inhibitors of AP-1 and NF-kappaB mediated transcriptional activations. The CoMFA model produced statistically significant results with the cross-validated q(2) of 0.594 and the conventional correlation coefficient r(2) of 0.968. The best CoMSIA model was obtained by the combination use of steric, electrostatic, hydrogen-bond donor and acceptor fields. The corresponding q(2) and r(2) of CoMSIA model were 0.703 and 0.932, respectively. From the cross-validated results, it can be seen that the CoMSIA model has a better predictive ability than CoMFA model due to the importance of the hydrogen bonds for the activity of these inhibitors. The predictive abilities of the two models were further validated by a test set of 15 compounds. The models gave predicted correlation coefficient r(pred)(2) of 0.891 for CoMFA model and 0.810 for CoMSIA model. Based on the above results, we identified the key structural features that may help to design potent inhibitors with improved activities: (1) the NH linker at the position R(4) acts as important hydrogen-bond donor and any group on phenyl or 2-thienyl ring of R(1) substituent decreases inhibitory activity, (2)further structural modification of compound 50 on the phenyl ring of the quinazoline ring considering steric, electrostatic and hydrogen-bond acceptor properties will influence the inhibitory activity. PMID- 19128858 TI - Association of matrix metalloproteinases with MRI indices of brain ischemia and aging. AB - Magnetic resonance imaging (MRI) findings of large white matter hyperintensities (LWMH), decreased brain volume and silent cerebral infarcts (SCI) are subclinical indices of brain ischemia and aging. Although the pathophysiology of these findings remains uncertain, extracellular matrix (ECM) remodeling, a process regulated by matrix metalloproteinases (MMPs) and their inhibitors (TIMPs), may be implicated. We evaluated the cross-sectional relations of circulating MMP-9 and TIMP-1 to these MRI indices in 583 stroke and dementia-free, Framingham Offspring participants (mean age 57 years, 58% women). Using multivariable regression MMP-9 (detectable versus non-detectable) and TIMP-1 (modeled as sex specific quartiles) were related to LWMH (>1S.D. above age-specific mean; yes/no), SCI (yes/no) and total brain volume (ratio of parenchymal to intracranial volume, TCBVr). Mean TCBVr was 0.78 (S.D. 0.03), 13% of subjects had LWMH and 12% had SCI. Detectable MMP-9 was associated with higher prevalence of LWMH (OR 2.09, 95%confidence interval (CI) 1.00-4.37), but not with TCBVr. TIMP-1 was associated with a high prevalence of LWMH (OR for Q4 versus Q1-3: 1.83, 95%CI 1.06-3.18) and with lower mean TCBVr (Q4 associated with 0.17 S.D. units lower value relative to Q1-3; p=0.04). Neither biomarker was associated with SCI. Our findings are preliminary but if confirmed in further studies, suggest a pathophysiological role for the MMP/TIMP pathway in processes of brain ischemia and aging. PMID- 19128860 TI - Comparative chemometric modeling of cytochrome 3A4 inhibitory activity of structurally diverse compounds using stepwise MLR, FA-MLR, PLS, GFA, G/PLS and ANN techniques. AB - Twenty-eight structurally diverse cytochrome 3A4 (CYP3A4) inhibitors have been subjected to quantitative structure-activity relationship (QSAR) studies. The analyses were performed with electronic, spatial, topological, and thermodynamic descriptors calculated using Cerius 2 version 10 software. The statistical tools used were linear [multiple linear regression with factor analysis as preprocessing step (FA-MLR), stepwise MLR, partial least squares (PLS), genetic function algorithm (GFA), genetic PLS (G/PLS)] and non-linear methods [artificial neural network (ANN)]. All the five linear modeling methods indicate the importance of n-octanol/water partition coefficient (logP) along with different topological and electronic parameters. The best model obtained from the training set (stepwise regression) based on highest external predictive R(2) value and lowest RMSEP value also showed good internal predictive power. Other models like FA-MLR, PLS, GFA and G/PLS are also of statistically significant internal and external validation characteristics. The best model [according to r(m)(2) for the test set, as defined by P.P. Roy, K. Roy, QSAR Comb. Sci. 27 (2008) 302-313] obtained from ANN showed a good r(2) value (determination coefficient between observed and predicted values) for the test set compounds, which was superior to those of other statistical models except the stepwise regression derived model. However, based upon the r(m)(2) value (test set), which penalizes a model for large differences between observed and predicted values, the stepwise MLR model was found to be inferior to other methods except PLS. Considering r(m)(2) value for the whole set, the G/PLS derived model appears to be the best predictive model for this data set. For choosing the best predictive model from among comparable models, r(m)(2) for the whole set calculated based on leave-one-out predicted values of the training set and model-derived predicted values for the test set compounds is suggested to be a good criterion. PMID- 19128861 TI - Synthesis and antitumor activity of optically active thiourea and their 2 aminobenzothiazole derivatives: a novel class of anticancer agents. AB - A novel series of optically active 2-aminobenzothiazole derivatives were synthesized by reaction of optically active amine (I) with thiophosgene to obtain optically active isothiocyanates (IIa-h) which on condensation with 4-fluoro-3 chloro aniline (III) yielded various optically active thioureas (IVa-h). Further oxidative cyclisation in the presence of bromine and chloroform yielded title compounds (Va-h). The structures of these compounds were established by IR, (1)H NMR, (13)C NMR, Mass and HRMS. The compounds (IVa-h and Va-h) were evaluated for in vitro cytotoxicity against mouse Ehrlich Ascites Carcinoma (EAC) and two human cancer cell lines (MCF-7 and HeLa). In preliminary MTT [3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide] cytotoxicity studies the optically active thiourea derivatives (IVe, IVf and IVh) were found most effective. In EAC cells the IC(50) values for IVe, IVf, IVh and Vg were found in the range of 10-24 microM, whereas in MCF-7 and HeLa cells the IC(50) values were observed in the range of 15-30 microM and 33-48 microM, respectively. In alkaline comet assay the compounds (IVe and IVf) showed dose-dependent DNA damaging activity. PMID- 19128862 TI - Study of the comparative dynamics of the incorporation of tissue free-water tritium (TFWT) in bulrushes (Typha latifolia) and carp (Cyprinus carpio) in the Almaraz nuclear power plant cooling reservoir. AB - The Almaraz nuclear power plant (Spain) uses the water of Arrocampo reservoir for cooling, and consequently raises the radioactive levels of the aquatic ecosystem of this reservoir. From July 2002 to June 2005, monthly samples of surface water, bulrushes (Typha latifolia) and carp (Cyprinus carpio) were collected from this reservoir. They were analyzed to determine the temporal evolution of the levels of (3)H in surface water and of its transfer from the surface water to free-water in the tissues (TFWT) of the aforementioned two organisms. The tritium levels in the surface water oscillate with a biannual period, with their values in the study period ranging between 53 and 433 Bq/L. The incorporation of tritium to bulrushes and carp was fairly similar, the respective mean concentration factors being 0.74 and 0.8 (unitless, as Bq/L tissue water per Bq/L reservoir water). The temporal evolution of the levels fairly closely followed that observed for the surface water tritium, although detailed analysis showed the dominant periodicity for the bulrushes to be annual. This difference reflects the influence on the incorporation of tritium to bulrushes of diverse environmental and metabolic factors, especially evapotranspiration and the seasonal growth of this plant. PMID- 19128863 TI - Support in the first stage of labour from a female relative: the first step in improving the quality of maternity services. AB - OBJECTIVE: to undertake a substantial descriptive study to assess whether the provision of labour support, in hospitals in Jordan, by a female relative during the first stage of labour affected duration of labour, use of pharmacological pain relief, mode of delivery and the woman's postpartum perception of the birth experience. DESIGN: a non-randomised comparison study. SETTING: maternity ward in Al-Karak Hospital, the main governmental hospital in the south region of Jordan. PARTICIPANTS: a convenience sample of 226 nulliparous women who had a single term fetus, were expecting an uncomplicated vaginal birth and were in spontaneous labour at the time of admission. INTERVENTION: supportive companionship from a female relative with no medical or nursing experience, concentrating on comfort, reassurance and praise. RESULTS: women who had support during labour were significantly less likely than those in the control group to have pharmacological pain relief, and were significantly more likely than those in the control group to report a good birth experience. There were no statistically significant differences between groups in mode of delivery or duration of labour. CONCLUSION: labour support by a female relative is a cost-effective and beneficial practice to apply to intrapartum care in hospitals in developing countries with limited resources, such as Jordan. Labour support is not routine practice and is not permitted in Jordan, as in many other countries. Consideration should be given to change maternity systems to ensure that all women have access to such support. PMID- 19128864 TI - A roadmap to rumination: a review of the definition, assessment, and conceptualization of this multifaceted construct. AB - Rumination has been widely studied and is a crucial component in the study of cognitive vulnerabilities to depression. However, rumination means different things in the context of different theories, and has not been uniformly defined or measured. This article aims to review models of rumination, as well as the various ways in which it is assessed. The models are compared and contrasted with respect to several important dimensions of rumination. Guidelines to consider in the selection of a model and measure of rumination are presented, and suggestions for the conceptualization of rumination are offered. In addition, rumination's relation to other similar constructs is evaluated. Finally, future directions for the study of ruminative phenomena are presented. It is hoped that this article will be a useful guide to those interested in studying the multi-faceted construct of rumination. PMID- 19128865 TI - Determinants of performance failure in the nursing home industry. AB - This study investigates the determinants of performance failure in U.S. nursing homes. The sample consisted of 91,168 surveys from 10,901 facilities included in the Online Survey Certification and Reporting system from 1996 to 2005. Failed performance was defined as termination from the Medicare and Medicaid programs. Determinants of performance failure were identified as core structural change (ownership change), peripheral change (related diversification), prior financial and quality of care performance, size and environmental shock (Medicaid case mix reimbursement and prospective payment system introduction). Additional control variables that could contribute to the likelihood of performance failure were included in a cross-sectional time series generalized estimating equation logistic regression model. Our results support the contention, derived from structural inertia theory, that where in an organization's structure change occurs determines whether it is adaptive or disruptive. In addition, while poor prior financial and quality performance and the introduction of case mix reimbursement increases the risk of failure, larger size is protective, decreasing the likelihood of performance failure. PMID- 19128866 TI - Smoking cessation in England: intentionality, anticipated ease of quitting and advice provision. AB - Smoking prevalence in England continues to reduce but further reduction is increasingly difficult. Cessation policy has successfully targeted those who want to quit but further reduction will need to shift attention to more difficult 'core smoker' populations. Following the established 'stages of change' perspective, this paper considers the characteristics of people who do not intend to quit smoking, anticipate difficulties in quitting and have not received advice about quitting. We deploy multilevel models of data drawn from the Health Survey for England years 2002-2004, and the NHS Primary Care Trust Patient Surveys for 2004 and 2005. It was found that variations in intentionality and anticipated ease of quitting are associated with individual factors such as smoking intensity, parental smoking, age/length of time as a smoker and the nature of the advice-giving consultation. Household composition and household income are also implicated in the intention to quit and anticipated difficulties in quitting. Once individual and household factors are taken into account the only identifiable area-level variation is reduced intentionality towards quitting in rural areas. We conclude by arguing that further gains in smoking cessation must focus on understanding the characteristics of 'hard-to-engage' populations. PMID- 19128867 TI - Incontinence and trauma: sexual violence, female genital cutting and proxy measures of gynecological fistula. AB - Obstetric fistula, characterized by urinary or fecal incontinence via the vagina, has begun to receive attention on the international public health agenda, however less attention has been given to traumatic fistula. Field reports indicate that trauma contributes to the burden of vaginal fistula, especially in regions wrought by civil unrest, however evidence is largely anecdotal or facility-based. This paper specifically examines the co-occurrence of incontinence and two potential sources of trauma: sexual violence and female genital cutting using the most recent Demographic and Health Surveys in Malawi, Rwanda, Uganda and Ethiopia. Multivariate selection models are used to control for sampling differences by country. Results indicate that sexual violence is a significant determinant of incontinence in Rwanda and Malawi, however not in Uganda. Simulations predict that elimination of sexual violence would result in from a 7 to a 40% reduction of the total burden of incontinence. In contrast, no evidence is found that female genital cutting contributes to incontinence and this finding is robust for types of cutting and high risk samples. Results point to the importance of reinforcing prevention programs which seek to address prevention of sexual violence and for the integration of services to better serve women experiencing both sexual violence and incontinence. PMID- 19128868 TI - The socio-economic conditioning of difficulties in adaptation to pregnancy following assisted reproductive techniques. AB - OBJECTIVES: The aim of the study was to analyse the demographic, social and economic conditioning of the adaptation process in women who became pregnant following assisted reproductive techniques. STUDY DESIGN: The analysis was performed with the use of a questionnaire, which was designed to evaluate, among other factors, the attitudes of women towards their pregnancies. The study group included 250 primiparae in whom pregnancy had resulted either from artificial insemination by their husband or a donor or from in vitro fertilisation. RESULTS: Two subgroups were identified. The first consisted of women who were well adapted to pregnancy (n=90, 36%), while the second included those subjects who experienced difficulties in adapting (n=160, 64%). The adaptation of women to pregnancy was significantly different (p<0.05) with regard to age, education and environment, as well as housing and material conditions, and, moreover, with regard to the duration of infertility treatment and the costs incurred. The type of assisted reproductive technique that could have initiated the pregnancy also turned out to be vital. Whether the women were in permanent employment was not significant (p>0.05). CONCLUSIONS: More difficulties in adapting to pregnancy as a result of assisted reproductive techniques were reported by younger women (up to 30 years of age), by those inhabiting a rural environment and by those who had had a better education or who were living in better socio-economic conditions. Moreover, those subjects who benefited from more expensive and more complicated procedures, and thus incurred higher treatment costs, also experienced substantial difficulties. PMID- 19128869 TI - Approximating the distribution of recreational visits from on-site survey data. AB - While convenient and often used, on-site surveys are biased by the fact that users who visit the site more often are proportionately more likely to be sampled. This so-called avidity or size biased sampling results in over estimating the visitation patterns of the average user. This analysis develops a rule of thumb method that may easily be applied by recreation site managers to visitation data collected on-site in order to infer behavior of the average user of the site. The key assumption that drives the derivation is that the visitation data of users is logarithmically distributed. To evaluate the methodology, we analyze several data sets of recreational users assuming that they reflect the populations of users and from these construct hypothetical on-site samples. PMID- 19128870 TI - Measuring the accuracy of agro-environmental indicators. AB - Numerous agro-environmental indicators have been developed by agronomists and ecologists during the last 20 years to assess the environmental impact of farmers' practices, and to monitor effects of agro-environmental policies. The objectives of this paper were (i) to measure the accuracy of a wide range of agro environmental indicators from experimental data and (ii) to discuss the value of different information typically used by these indicators, i.e. information on farmers' practices, and on plant and soil characteristics. Four series of indicators were considered in this paper: indicators of habitat quality for grassland bird species, indicators of risk of disease in oilseed rape crops, indicators of risk of pollution by nitrogen fertilizer, and indicators of weed infestation. Several datasets were used to measure their accuracy in cultivated plots and in grasslands. The sensitivity, specificity, and probability of correctly ranking plots were estimated for each indicator. Our results showed that the indicators had widely varying levels of accuracy. Some show very poor performance and had no discriminatory ability. Other indicators were informative and performed better than random decisions. Among the tested indicators, the best ones were those using information on plant characteristics such as grass height, fraction of diseased flowers, or crop yield. The statistical method applied in this paper could support researchers, farm advisers, and decision makers in comparing various indicators. PMID- 19128871 TI - Leptomeningeal dissemination of low-grade intramedullary gliomas: about one case and review. AB - Leptomeningeal (LM) dissemination arises rarely in patients with low-grade gliomas. Few cases of LM dissemination of spinal cord low-grade gliomas have been reported in children. Here, we report a case of an intramedullary spinal cord low grade glioma occurring in an adult presenting by LM dissemination. A 57-year-old man presented with a history of headaches and visual acuity deterioration. Neurological examination revealed mental confusion, decreasing left limb strength and left positive Babinski sign. Lumbar puncture showed low CSF glucose, high CSF protein and no tumor cells. Spinal MRI revealed an intramedullary spinal lesion with a diffuse leptomeningeal enhancement. A spinal meningeal biopsy showed arachnoiditis with isolated atypical cells which were cytokeratin immunopositive. A differential diagnosis was proposed between arachnoiditis close to an infectious process or close to meningeal carcinomatosis. An intramedullary biopsy was programmed but the patient died 1 month after biopsy. The post-mortem examination showed thickened leptomeninges and an ill-circumscribed intramedullary grey lesion. Histological examination revealed a low-grade glial neoplasm composed of monomorphous spindle "piloid" cells with diffuse dissemination to leptomeninges. Glial fibrillary acidic protein immunostaining showed scattered immunoreactive cells. Clinical and pathological features of this case are unusual. Pathogenesis and prognosis of low-grade gliomas with LM dissemination are poorly understood. This case, like cases occurring in children, cannot easily be classified in the present WHO system of classification of CNS tumors. Spinal low-grade gliomas with LM dissemination may represent a possible distinctive clinico-pathological entity. PMID- 19128872 TI - Posterior reversible encephalopathy syndrome as a complication of acute lupus activity. AB - OBJECTIVES: We aimed to describe the clinical and imaging characteristics; associated risk factors and neurological outcome of posterior reversible encephalopathy syndrome (PRES) in patients with systemic lupus erythematosus (SLE). METHODS: From October 2001 to January 2007, we identified patients with SLE and the criteria for PRES in our institution, which is a tertiary-care referral center for patients with SLE; the patients were evaluated at baseline and followed to determine the clinical outcome. RESULTS: We identified 22 episodes of PRES in 21 patients; 20 (95.2%) were women, mean age of onset was 24.9+/-8.6 years, all patients had high systemic activity (SLEDAI scores from 12 to 39). Acute hypertension was observed in 18 episodes (81.8%), and renal failure in 16 (72.7%); only 3 patients were on cyclophosphamide at the time of the onset of PRES. Persistent neurological deficit was observed in 2 cases; one patient died during the acute episode. CONCLUSIONS: PRES is a central nervous system syndrome that is observed in SLE patients. It was associated mainly to high systemic activity, acute hypertension, and renal failure. Although reversibility is common, residual neurological damage may be observed. PMID- 19128873 TI - Rapid decolorization of azo dye methyl orange in aqueous solution by nanoscale zerovalent iron particles. AB - Azo dyes are recalcitrant and refractory pollutants that constitute a significant burden on the environment. The report here is focused on the decolorization treatment of water soluble azo dye methyl orange (MO) by chemically synthesized nanoscale zerovalent iron (NZVI) particles. Experimental variables such as initial dye concentration, iron dosage, solution pH and temperature were studied systematically. Batch experiments suggest that the decolorization efficiency was enhanced with the increase of NZVI dosage and reaction temperature, but decreased with increasing initial dye concentration and initial solution pH. Further studies indicated that existence of inorganic salt (Na(2)SO(4)) could inhibit the decolorization of MO. Kinetic analyses based on the experimental data elucidated that the decolorization process followed a first order exponential decay kinetics model. The activation energy was determined to be 35.9 kJ/mol. PMID- 19128874 TI - Simultaneous removals of NOx, HC and PM from diesel exhaust emissions by dielectric barrier discharges. AB - The main target of this work is to characterize the abatements of particulate matter (PM), hydrocarbons (HC) and nitrogen oxides (NO(x)) from an actual diesel exhaust using dielectric barrier discharge technology (DBD). The effects of several parameters, such as peak voltage, frequency and engine load, on the contaminant removals have been investigated intensively. The present study shows that for a given frequency, the removals of PM and HC are enhanced with the increase of peak voltage and level off at higher voltage, while in the range of higher voltages a decline of NO(x) removal efficiency is observed. For a given voltage, the maximums of specific energy density (SED) and removal efficiency are attained at resonance point. The increase of peak voltage will result in a significant decrease of energy utilization efficiency of DBD at most engine loads. Alkanes in soluble organic fraction (SOF) are more readily subjected to removals than polycyclic aromatic hydrocarbons (PAHs). PMID- 19128875 TI - Radiation induced degradation of dyes--an overview. AB - Synthetic dyes are a major part of our life. Products ranging from clothes to leather accessories to furniture all depend on extensive use of organic dyes. An unfortunate side effect of extensive use of these chemicals is that huge amounts of these potentially carcinogenic compounds enter our water supplies. Various advanced oxidation processes (AOPs) including the use of high-energy radiation have been developed to degrade these compounds. In this review, dye decoloration and degradation as a result of its exposure to high energy radiation such as gamma radiation and pulsed electron beam are discussed in detail. The role of various transient species such as H, OH and e(aq)(-) are taken into account as reported by various researchers. Literature citations in this area show that e(aq)(-) is very effective in decolorization but is less active in the further degradation of the products formed. The degradation of the dyes is initiated exclusively by OH attack on electron-rich sites of the dye molecules. Additionally, various parameters that affect the efficiency of radiation induced degradation of dyes, such as effect of radiation dose, oxygen, pH, hydrogen peroxide, added ions and dye classes are also reviewed and summarized. Lastly, pilot plant application of radiation for wastewater treatment is briefly discussed. PMID- 19128876 TI - Application of accelerated carbonation with a combination of Na2CO3 and CO2 in cement-based solidification/stabilization of heavy metal-bearing sediment. AB - The efficient remediation of heavy metal-bearing sediment has been one of top priorities of ecosystem protection. Cement-based solidification/stabilization (s/s) is an option for reducing the mobility of heavy metals in the sediment and the subsequent hazard for human beings and animals. This work uses sodium carbonate as an internal carbon source of accelerated carbonation and gaseous CO(2) as an external carbon source to overcome deleterious effects of heavy metals on strength development and improve the effectiveness of s/s of heavy metal-bearing sediment. In addition to the compressive strength and porosity measurements, leaching tests followed the Chinese solid waste extraction procedure for leaching toxicity - sulfuric acid and nitric acid method (HJ/T299 2007), German leaching procedure (DIN38414-S4) and US toxicity characteristic leaching procedures (TCLP) have been conducted. The experimental results indicated that the solidified sediment by accelerated carbonation was capable of reaching all performance criteria for the disposal at a Portland cement dosage of 10 wt.% and a solid/water ratio of 1:1. The concentrations of mercury and other heavy metals in the leachates were below 0.10mg/L and 5mg/L, respectively, complying with Chinese regulatory level (GB5085-2007). Compared to the hydration, accelerated carbonation improved the compressive strength of the solidified sediment by more than 100% and reduced leaching concentrations of heavy metals significantly. It is considered that accelerated carbonation technology with a combination of Na(2)CO(3) and CO(2) may practically apply to cement-based s/s of heavy metal-bearing sediment. PMID- 19128877 TI - The photooxidative destruction of C.I. Basic Yellow 2 using UV/S2O8(2-) process in a rectangular continuous photoreactor. AB - The photooxidative decolorization of C.I. Basic Yellow 2 (BY2), was investigated using UV radiation in the presence of peroxydisulfate (S(2)O(8)(2-)) in a rectangular photoreactor at experimental condition. S(2)O(8)(2-) and UV-light showed negligible effect when they were used independently. Removal efficiency of BY2 was sensitive to the operational parameters such as initial concentrations of S(2)O(8)(2-), BY2, light intensity, flow rate and pH. The conversion ratios of BY2 at the volumetric flow rates of 330, 500 and 650 ml/min were 84%, 79%, 51% in 30 min, respectively. Our results showed that light intensity was a beneficial parameter for dye removal. The results showed that in the presence of S(2)O(8)(2 ), the photooxidation quantum yield obtained was higher than direct photolysis quantum yield, suggesting that photodecay of BY2 was dominated by photooxidation. The electrical energy per order (E(EO)) values for decolorization of BY2 solution was calculated. Results show that applying a desired peroxydisulfate concentration can reduce the E(EO). PMID- 19128878 TI - Activated carbons prepared from wood particleboard wastes: characterisation and phenol adsorption capacities. AB - The problems of valorisation of particleboard wastes on one hand, and contamination of aqueous effluents by phenolic compounds on the other hand, are simultaneously considered in this work. Preparation of activated carbons from a two steps thermo-chemical process, formerly designed for generating combustible gases, is suggested. The resultant carbonaceous residue is activated with steam at 800 degrees C. Depending on the preparation conditions, surface areas within the range 800-1300 m(2)/g are obtained, close to that of a commercial activated carbon (CAC) specially designed for water treatment and used as a reference material. The present work shows that particleboard waste-derived activated carbons (WAC) are efficient adsorbents for the removal of phenol from aqueous solutions, with maximum measured capacities close to 500 mg/g. However, most of times, the adsorption capacities are slightly lower than that of the commercial material in the same conditions, i.e., at equilibrium phenol concentrations below 300 ppm. Given the extremely low cost of activated carbons prepared from particleboard waste, it should not be a problem to use it in somewhat higher amounts than what is required with a more expensive commercial material. Phenol adsorption isotherms at 298 K were correctly fitted by various equations modelling type I and type II isotherms for CAC and WAC, respectively. Phenol adsorption isotherms of type II were justified by a 3-stages adsorption mechanism. PMID- 19128879 TI - Ketamine for chronic non-cancer pain. PMID- 19128880 TI - Mechanisms of analgesia by gabapentin and pregabalin--calcium channel alpha2 delta [Cavalpha2-delta] ligands. PMID- 19128881 TI - Pain free and awake to enjoy it! PMID- 19128882 TI - Radiography for the diagnosis of spirocercosis in apparently healthy dogs, St. Kitts, West Indies. AB - Fifty apparently healthy island dogs presenting to the Ross University School of Veterinary Medicine (RUSVM), St. Kitts, West Indies for neutering were used in this prospective study. Twelve of the dogs (24%) were diagnosed with spirocercosis based on a positive fecal analysis and characteristic lesions seen during esophagoscopy. Routine thoracic survey radiographs revealed changes previously reported with spirocercosis in 10/12 (sensitivity=83%) infected dogs, but in none of the uninfected dogs (38/38; specificity=100%). The most common radiographic changes were an increased fluid density within the caudal dorsal thorax on the lateral view and a widening and/or bulging of the caudal mediastinum on the dorsoventral view. After oral administration of barium sulfate, barium retention or a tortuous esophagus was visible in all infected dogs (12/12; sensitivity 100%) and in one uninfected dog (1/38; specificity 97%). The results show spirocercosis is common on St. Kitts and that radiographs are as dependable as fecal analysis and/or endoscopy in diagnosing the condition. PMID- 19128883 TI - Effect of storage media, temperature, and time on preservation of Cryptosporidium parvum oocysts for PCR analysis. AB - The effect of storage media, temperature, and time on suitability of oocysts for use in subsequent molecular studies was examined. Cryptosporidium parvum oocysts were stored for 3, 6, 9, or 12 months in sterile dH(2)O, 70 or 95% ethanol, (room temperature [RT], 4, -20, and -70 degrees C), 10% formalin (RT and 4 degrees C), PBS, TE buffer, antibiotic-antimycotic (A-A) solution (4, -20 and -70 degrees C), 2% sulphuric acid, 2.5% potassium dichromate (4 degrees C), and gDNA from 10(4) oocysts was extracted in triplicate and subjected to PCR. To determine the effect of storage media on PCR sensitivity, gDNA from 10(4), 10(2), and 10(0) oocysts stored for 15 months in the media listed above at RT or 4 degrees C was also extracted in triplicate and subjected to PCR. At RT, ethanol was suitable for up to 15 months, while gDNA from oocysts stored in dH(2)O amplified inconsistently after 3 months. At 4 degrees C, all tested media except dH(2)O and formalin were suitable for storage of 10(4) oocysts up to 15 months, but only 70% ethanol, A-A solution, 2% sulphuric acid and 2.5% potassium dichromate supported amplification of gDNA from fewer than 100 oocysts. At -20 degrees C, 95% ethanol, PBS, or TE were suitable for up to 9 months, while 70% ethanol and A-A solution were effective up to 12 months, and gDNA from oocysts stored in dH(2)O was inconsistently amplified after 6 months. Storage at -70 degrees C for up to 12 months was effective regardless of media type. Oocysts stored in formalin at RT or 4 degrees C could not be amplified by PCR despite washing prior to gDNA extraction. To maintain gDNA quality suitable for PCR, it is recommended that coccidian oocysts be stored at -70 degrees C in dH(2)O, ethanol, PBS, TE or A-A solution, at 4 degrees C in A-A or ethanol, or at RT in ethanol where refrigerated storage is unavailable. PMID- 19128884 TI - Early neglect and abuse predict diurnal cortisol patterns in adults A study of international adoptees. AB - Neglect and abuse early in life have been associated with increased and decreased cortisol levels, and also with an altered diurnal cortisol slope. In the present study, we investigated the long-term relationship between early maltreatment - at different levels of severity - and basal cortisol secretion in adults adopted as children. A sample of international adoptees was followed from childhood to adulthood. In childhood, adoptive parents had provided information about neglect and abuse prior to adoption. As adults, adoptees collected saliva samples four times a day. The relationship between early maltreatment and cortisol secretion was examined, primarily with multilevel analyses in 623 adoptees. Morning cortisol levels were lower in adoptees whose adoptive parents had reported severe neglect or abuse than in non-neglected or non-abused participants (respective estimates (standard errors (SEs)) and p-values: -0.33 (0.090), p=0.0002 and -0.63 (0.20), p=0.002). Relative to non-neglected adoptees, those who had allegedly experienced severe neglect also had a flatter diurnal slope (estimate (SE) and p value: 0.028 (0.0088), p=0.002). In contrast, relative to non-abused participants, adoptees whose reported abuse was moderately severe had high cortisol levels and a steeper cortisol diurnal slope (respective estimates (SEs) and p-values: 0.29 (0.13), p=0.003 and -0.039 (0.012), p=0.01). Thus, early neglect and abuse appear to have associations with cortisol levels and the diurnal slope, even when children are raised in another environment after their early maltreatment. Our study suggests that the severity of the early maltreatment may be related to the basal cortisol pattern. PMID- 19128885 TI - Serotonin transporter polymorphism predicts waking cortisol in young girls. AB - Major Depressive Disorder (MDD) is one of the most prevalent and costly of all psychiatric disorders. The hypothalamic pituitary adrenal (HPA)-axis, which regulates the hormonal response to stress, has been found to be disrupted in depression. HPA dysregulation may represent an important risk factor for depression. To examine a possible genetic underpinning of this risk factor without the confound of current or lifetime depression, we genotyped 84 never disordered young girls, over a third of whom were at elevated risk for depression, to assess the association between a polymorphism in the promoter region of the serotonin transporter (5-HTT) gene and diurnal variation in HPA axis activity. This 5-HTT-linked polymorphic region (5-HTTLPR) has been previously found to interact with stress to increase risk for depression. We found 5-HTTLPR to be significantly associated with diurnal cortisol levels: girls who were homozygous for the short-allele had higher levels of waking (but not afternoon or evening) cortisol than did their long-allele counterparts. This finding suggests that genetic susceptibility to HPA-axis dysregulation, especially apparent in levels of waking cortisol, is detectable in individuals as young as 9 years of age. PMID- 19128886 TI - The cortisol awakening response, seasonality, stress and arousal: a study of trait and state influences. AB - The aim of this study was to investigate the relationship between the cortisol awakening response (CAR) and two key state variables (morning stress and arousal) and the trait-like variable of seasonality as recent evidence suggests that the CAR is subject to both state and trait influences. The CAR was examined across two consecutive winter days in 50 healthy participants. Participants collected saliva samples in the domestic setting immediately on awakening, then at 15, 30 and 45min post-awakening on the two study days. Concomitant trait and state measures were examined, notably seasonal changeability in mood as a trait, and self-reported stress and arousal as state measures. Although there was correlational stability for measures of the CAR across days, there was a significant difference in the magnitude of the increase in cortisol levels following awakening between the two study days, being greater on the first sampling day. This reduction in the magnitude of cortisol increase was significantly associated with an observed reduction across the 2 days in self reported arousal assessed at 45min following awakening. Participants reported greater arousal (more alert, active, energetic and stimulated, less drowsy, tired and sluggish) on the first study day than the second. Average CAR across days was associated with seasonality score, greater propensity for seasonal changes in mood being associated with smaller average CAR. High seasonality scorers were also more likely as a group to show a strong association between daily changes in state arousal and CAR. This study supports the view that the CAR is, in part, susceptible to short-term changes in state variables, notably perceived arousal, while observing a novel link between CAR and the trait variable of perceived seasonality. Finally a tentative finding suggests the importance of examining for possible interaction between trait and state effects, evidenced by a significantly greater association between state arousal changes and cortisol response changes in those with high (trait) seasonality. PMID- 19128887 TI - Intensified testing for attention-deficit hyperactivity disorder (ADHD) in girls should reduce depression and smoking in adult females and the prevalence of ADHD in the longterm. AB - Attention-deficit hyperactivity disorder (ADHD) is the most common neurobehavioral disorder in youth. About a third to one-half of the affected subjects continue to have symptoms in adulthood. Remarkably, the prevalence numbers published for adult females are higher than for girls. The differences in the epidemiological data between the age groups clearly point to underdiagnosed ADHD in girls. Major depression, the most frequent psychiatric condition worldwide in adulthood, is twice as common in female as in male adults. Anxiety and depression are also among the most common comorbidities in adults with ADHD. Therefore, an undiagnosed ADHD may often underlie the psychopathology in depressive women. Another possibly associated phenomenon is the increased frequency of smoking in adult females. Since nicotine indirectly enhances the intrasynaptic dopamine level which presumably is too low both in ADHD and in depression, smoking might be used as a self-medication in women with untreated ADHD and consecutive depression. Furthermore, smoking during pregnancy is a major risk factor for ADHD in the offspring, so the vicious circle is complete. Depression in mothers of children with ADHD is associated with a higher rate of comorbidity in the children. Improved screening for ADHD in girls and treatment in childhood might thus reduce the rate of depression and smoking in adult females. We hypothesize that earlier identification and interventions might not only improve the lives of millions of girls and women but might also reduce the prevalence rates in future generations or at least moderate the deviant behaviour in this highly heritable disorder in which the development and severity of symptoms and the functional impairment depend to a high degree on epigenetic factors. PMID- 19128888 TI - Erythropoietin as a novel therapeutic agent for atrophic age-related macular degeneration. AB - The purpose of this article is to propose a novel therapeutic approach to the treatment of age-related macular degeneration (ARMD), the leading cause of blindness in the elderly population (over 60 years of age) in developed countries. Although recent advances have been made in the treatment of the neovascular form of ARMD, there is still no effective treatment for the most prevalent atrophic form of ARMD. Although the exact etiology and molecular pathogenesis of the atrophic ARMD are not fully understood, it is believed that oxidative stress and local inflammation play a major role in the pathologic processes and that the disease is triggered by dysfunction in the retinal pigment epithelia, leading to the degeneration of macular photoreceptor cells, followed by irreversible loss of vision. Considering that erythropoietin (EPO) has antioxidant, anti-inflammatory, and neuroprotective properties, we hypothesize that it can be developed as a novel therapeutic agent for the treatment of the atrophic form of ARMD. Future studies are needed to confirm or rule out this hypothesis. If successful, such studies may also help shield the lights on molecular mechanisms of atrophic ARMD. PMID- 19128889 TI - Hurtle cell change: the precursor of the immunologic phase of thyroiditis. PMID- 19128890 TI - Gas bubbles may not be the underlying cause of decompression illness - The at depth endothelial dysfunction hypothesis. AB - Gas formed in tissues and the circulating blood due to decompression is thought to be a significant factor in the progression of decompression illness (DCI). DCI is a potential problem for a growing population of professional and recreational divers. We hypothesise that these gas bubbles are not the causative agent in progression of DCI, rather an exacerbating factor. Endothelial dysfunction caused by a temporary loss of haemostasis due to increased total oxidant status is postulated to be the cause in this at-depth endothelial dysfunction hypothesis. Breathing oxygen at any pressure increases the oxidant status in the circulation causing vasoconstriction; this increase can be prevented by antioxidants, such as Vitamin C, maintaining haemostasis and preventing activation of endothelium, leukocyte recruitment and subsequent localised inflammation. Bubbles have the potential to exacerbate the situation on decompression by damaging the vascular endothelium either through ischemia/reperfusion, physical contact with the endothelium or by an increase in shear stress. Furthermore, this damage may manifest itself in the release of endothelial membrane fragments (microparticles). PMID- 19128891 TI - Combination sonodynamic therapy with immunoadjuvant may be a promising new modality for cancer treatment. AB - Sonodynamic therapy (SDT) is a new cancer therapy basing on photodynamic therapy (PDT). Some chemicals produce free radicals on irradiation with laser (photosensitizers) or ultrasound (sonosensitizers). These active molecules destroy biological tissues, thus producing therapeutic effects. Although PDT has been adopted in clinical cancer therapy especially for superficial cancers, this modality is under continued investigation for improved efficacy and expanded use. For example, PDT-generated tumor cell lysates are effective cancer vaccines; treatment of PDT in conjunction with immunoadjuvant, called "PDT-immunoadjuvant therapy" (PIT), "photoimmunotherapy" or "laser immunotherapy", is considered to be a promising therapeutic interventions for the treatment of cancers. Ultrasound, especially focused ultrasound, can penetrate deeply into tissues and can be focused into a small region of a tumor to activate the cytotoxicity of sonosensitizers. This is a unique advantage in the non-invasive treatment of nonsuperficial tumors when compared to laser light used for PDT. For the similar mechanism of PDT and SDT, we hypothesize that SDT may be exploited for the generation of effective therapeutic cancer vaccines like PDT; and combination SDT with Immunoadjuvant may be a promising systemic treatment modality, not only for superficial cancers but also for deep-seated tumors, which would surpass PIT. PMID- 19128892 TI - Shower emboli and digital necrosis after a single corticosteroid injection for trigger thumb: case report. AB - Local corticosteroid injection into the hand is the preferred initial step in the management of trigger finger owing to its safety and efficacy. We report the case of a patient with shower emboli and digital necrosis after a local corticosteroid injection for the treatment of trigger thumb. Given the patient's subsequent negative hypercoagulability workup, we suspect that the patient's symptoms resulted from the injection of steroids into the princeps pollicis artery, with subsequent retrograde flow into multiple other arteries of the hand. The patient was managed conservatively and ultimately her symptoms resolved. No new areas of digital necrosis have appeared in 8 years of follow-up. PMID- 19128894 TI - Detection and characterization of bovine torovirus from the respiratory tract in Japanese cattle. AB - Bovine torovirus (BToV), a member of the Coronaviridae family, is a causative agent of diarrhea in cattle, but it may also possess tropism for the respiratory tract. However, no surveys concerning with the relation between respiratory symptoms and the detection of BToV have been conducted in wide range. Among 311 nasal samples, BToV gene products were detected in seven samples (rBToV-1 to -7) derived only from calves with respiratory symptoms, suggesting that BToV may be a predisposing factor and/or causative agent for bovine respiratory disease. Regarding the degree of similarity between the spike and hemagglutinin-esterase coding regions, the rBToVs showed over 90.8% similarity with one another and 73.5 99.0% similarity with fecal tract-derived BToVs. rBToV-1, -2, and -3 were identical despite their being collected during different seasons; in comparison, rBToV-4 and -5 were distinct despite the fact that they were collected from the same herd, suggesting the existence of diversity among domestic rBToVs. One animal with a BToV-positive nasal sample also shed the virus in its feces, suggesting dual tropisms for BToV. PMID- 19128893 TI - Longitudinal quantification of Ehrlichia canis in experimental infection with comparison to natural infection. AB - Ehrlichia canis is a major tick-borne bacterial pathogen of dogs. Quantitative real-time PCR was evaluated for the detection of E. canis in naturally (NI) and experimentally infected (EI) dogs. DNA was extracted from blood, spleen and conjunctival swabs of experimentally infected dogs pre- and post-infection (PI), and during doxycycline therapy, and from blood and conjunctivas of naturally infected dogs. The primers and probe were designed to amplify a 93bp fragment of the single copy E. canis 16S rRNA gene with the TaqMan system. All EI dogs were positive for E. canis DNA by 7d PI and developed clinical ehrlichiosis by 9-12d PI. A rapid increase in ehrlichial DNA in EI dogs correlated with the appearance of severe clinical signs of disease. The mean spleen and blood DNA copies significantly increased by more than 10-folds from 7d PI to 10 and 12d PI (p<0.05). E. canis DNA was undetectable in the blood by day 9 post-treatment. Although the spleen was more frequently positive than blood (15/15 specimens vs. 13/15), no significant differences were found between the mean ehrlichial DNA copies in the spleen and blood on each day of examination. In 12 naturally infected dogs, the mean blood DNA copies was similar to the number found in EI 7d PI, but significantly lower than the means of 10 and 12d PI (p<0.0001). Although the conjunctivas of all EI dogs were positive by 12d PI, only 3/5 (60%) NI dogs were positive also by conjunctival PCR. In conclusion, the kinetics of E. canis during acute experimental infection with complete pathogen clearance following doxycyline treatment was demonstrated for the first time by real-time PCR. The value of real-time PCR was shown in NI dogs as well as in EI dogs with spleen and blood sampling more sensitive than non-invasive conjunctival PCR. PMID- 19128895 TI - Experimental reproduction of bluetongue virus serotype 8 clinical disease in calves. AB - Cattle are commonly subclinically infected following natural or experimental infection with bluetongue virus (BTV). The introduction of BTV serotype 8 (BTV-8) in Europe has been characterized by the manifestation of clinical signs in infected cattle. In order to study the pathogenesis of BTV-8 in this host, an animal model able to reproduce the clinical manifestations of the disease is required. In this work, two calves were subcutaneously and intravenously injected with a low passage cell-adapted strain of BTV-8. Both calves showed typical bluetongue clinical signs, including pyrexia, ocular discharge, conjunctivitis, oral mucosal congestion, development of ulcers and necrotic lesions on the lips and tongue, submandibular oedema, coronitis and oedema of the coronet and pastern region. A score was assigned depending on the severity of the lesions and a total clinical score was calculated for each animal daily and at the end of the experiment. Both calves became viraemic 24h post-infection and seroconversion occurred between 7 and 11 days P.I. In this study we present the development of a protocol of infection in calves able to reproduce the severity of the lesions observed with BTV-8 in field conditions. PMID- 19128896 TI - Seroprevalence and risk factors for the presence of ruminant pestiviruses in the Dutch swine population. AB - Swine can be infected with classical swine fever virus (CSFV), as well as ruminant pestiviruses: bovine viral diarrhoea virus (BVDV), and Border disease virus (BDV). Cross-reactions between pestiviruses occur, both regarding protective immunity and in diagnostic tests. The presence of BVDV and BDV in a swine population may thus affect the transmission of CSFV, but also the diagnosis of a CSFV infection. In this study, the seroprevalence against BVDV and BDV in two categories of swine, sows and finishing pigs, in the Netherlands was determined. Furthermore, several risk factors, associated with the presence of swine and ruminants on the same farm or in the immediate surroundings, were evaluated. In sows, the seroprevalence against BVDV was 2.5% on the animal level, and 11.0% on herd level. In finishing pigs these prevalences were 0.42% and 3.2%, respectively. Antibodies against BDV were found in three sows only. Risk factors, associated with a BVDV-seropositive status in breeding pigs, were the presence of cattle on the same premises and a high density of sheep and/or goats herds in a radius of 3km. While BVDV and BDV hardly pose any threat to the swine population themselves, knowledge, and therefore regular monitoring, on the presence of these viruses in the swine population is important with respect to CSF eradication. It will allow for a better interpretation of diagnostic test results, both in terms of possible false positives and false negatives, but may also bring about additional measures or surveillance protocols in times of CSF outbreaks to avoid surprises caused by cross-reactivity with ruminant pestiviruses. PMID- 19128897 TI - Changes in antimicrobial resistance among Salmonella enterica subspecies enterica serovar Pullorum isolates in China from 1962 to 2007. AB - There are few data available for the trends of antimicrobial resistance of Salmonella enterica subspecies enterica serovar Pullorum (S. Pullorum) in China and other parts of the world. Thus, the objective of this study was to evaluate the changes in antimicrobial resistance of S. Pullorum isolated from diseased chickens in China from 1962 to 2007. A total of 450 S. Pullorum isolates were tested for their susceptibility to 17 antimicrobials in a disk diffusion method. 39-95% of the isolates displayed a high level of resistance, particularly against ampicillin, carbenicillin, streptomycin, tetracycline, trimethoprim and sulfafurazole. Isolates exhibited increased resistance to carbenicillin, spectinomycin, trimethoprim, trimethoprim/sulfamethoxazole and nalidixic acid during the study period. Moreover, 56.2% of the isolates exhibited multiple drug resistance (MDR; resistance> or =4 antimicrobials) and showed an increasing trend between 1970-1979 and 2000-2007. Therefore, the results suggest that certain measures, including continued surveillance of antimicrobial resistance and the rational use of antimicrobials, are necessary and important in order to control the rapid increase in antimicrobial resistance in S. Pullorum. PMID- 19128898 TI - Modulations of phenotype and cytokine expression of porcine bone marrow-derived dendritic cells by porcine reproductive and respiratory syndrome virus. AB - Phenotypic and functional property changes of bone marrow-derived immature dendritic cells (BM-imDCs) after porcine reproductive and respiratory syndrome virus (PRRSV) infection have been detailed in a previous report. A down-regulated expression of MHC I molecules along with an up-regulated expression of CD80/86 were observed in BM-imDCs after the exposure to PRRSV. In this study, we further investigate the expression of surface phenotypes of BM-imDCs in relation to their infection status. Exposure of PRRSV to BM-imDCs resulted in a down-regulated expression of MHC I and an up-regulated expression of CD80/86 in infected cells, as demonstrated by significant alterations in both percentage of expressing cells and mean fluorescence intensity (MFI) in PRRSV-positive cells. A significant suppression in MFI of MHC I and an increase in percentage of cells expressing CD80/86 were observed in noninfected bystander cells. We also demonstrated that exposure of BM-imDCs to PRRSV resulted in a significantly increased secretion of IL-1, IL-6, IL-8, IL-10 and IFN-gamma but not IL-12 or TNF-alpha. In addition, the PRRSV infection modulates cytokine expressions of BM-imDCs through their response to microbial pathogen-associated molecular patterns. These results will prove helpful in clarification of the factors that mediate host defense against PRRSV, as well as the possible interaction mechanisms between PRRSV and other microbes in the pathogenesis of PRRSV infection in pigs. PMID- 19128899 TI - Development of a loop-mediated isothermal amplification assay for sensitive and rapid detection of Campylobacter fetus. AB - We developed a sensitive and rapid loop-mediated isothermal amplification (LAMP) assay for detection of Campylobacter fetus. This assay provides simpler and more rapid detection of C. fetus than conventional biochemical and PCR assays. The assay correctly detected 60 C. fetus strains but not 55 non-fetusCampylobacter and 30 non-Campylobacter strains. The sensitivity of the LAMP assay in pure cultures and in a spiked bovine liver specimen was 10-fold more sensitive than that of the conventional PCR assay. The sensitivity of the LAMP assay in a spiked bovine vaginal mucus specimen was similar to that of the conventional PCR assay. The assay was markedly faster, requiring less than 40min for detection of C. fetus in a single colony on blood agar and 80min in spiked bovine specimens from the beginning of DNA extraction to final determination. Our LAMP assay is a simple and practical tool for detection of C. fetus regardless of subspecies. PMID- 19128900 TI - Genetic analysis of antimicrobial resistance in Escherichia coli isolated from diarrheic neonatal calves. AB - This study was carried out to screen and analyze the genetic basis of antimicrobial resistance in Escherichia coli strains isolated from neonatal calf diarrhea in Egypt. A total of 182 isolates of E. coli recovered from 91 diarrheic neonatal calves were analyzed for antimicrobial susceptibilities, the presence of class 1 and class 2 integrons and antimicrobial resistance genes. Nineteen isolates (10.4%) showed multidrug resistance phenotypes and harbored at least three antimicrobial resistance genes. PCR screening detected class 1 integrons in 19 isolates (10.4%) and class 2 integrons in 2 isolates (1.1%). The identified antimicrobial resistance genes within class 1 integrons were dihydrofolate reductase types: dfrA1, dfrA12, dfrA15 and dfrA17, which confer resistance to trimethoprim; aminoglycoside adenyltransferase types: aadA1, aadA2, aadA5, aadA7 and aadA23, which confer resistance to streptomycin and spectinomycin; and aminoglycoside acetyltransferase gene, aac(3)-Id, which confers resistance to gentamicin and sisomicin. Furthermore, many beta-lactamases encoding genes, plasmid-mediated quinolone resistance genes and florfenicol resistance gene were identified in this study. To the best of our knowledge, this is the first report for molecular characterization of antimicrobial resistance in E. coli isolated from diarrheic neonatal calves in Africa. PMID- 19128901 TI - Clinical aspects of induced hypothermia in full term neonates with perinatal asphyxia. AB - Moderate hypothermia is a novel neuroprotective therapy for full term neonates with severe perinatal asphyxia. Although the therapy appears to be safe, admission to a level III neonatal intensive care unit of these patients is justified. Potential complications include hypotension, tube obstruction due to sticky secretions, severe bradycardia, and thrombocytopenia. Furthermore, doses of commonly used drugs such as sedatives, anticonvulsants and antibiotics should be adjusted during hypothermia and on rewarming, and should be monitored carefully. Further studies aiming at optimizing onset, duration, and depth of hypothermia in neonates are necessary. Combination of hypothermia with drugs may further improve neuroprotection in asphyxiated full term neonates. PMID- 19128902 TI - Suppression of testicular function and sexual behavior by vaccination against GnRH (Equity) in the adult stallion. AB - The aim of this study was to evaluate the effects of an anti-GnRH vaccine on testosterone concentration, antibody titer, scrotal width, semen quality and sexual behavior in the stallion. Adverse reactions to the vaccine were also determined. Eight clinically healthy sexually experienced stallions aged between 6 and 15 years from the National Stud in Avenches (Switzerland) were used. Five stallions were immunized 3 times at an interval of 4 and 8 weeks, respectively, with 200 microg of a GnRH-protein conjugate (Equity, CSL Limited, Australia) intramuscularly in the neck and 3 control animals received an equivalent volume of saline solution. Plasma testosterone concentrations and GnRH antibody titers as well as semen quality and libido were determined weekly during 1 year (52 weeks). In addition, scrotal width was measured in all stallions before and 4, 8 as well as 12 months after first vaccination. Our results demonstrate that in 4 stallions plasma testosterone started to decrease after the second vaccination and remained suppressed for at least 6 months whereas one stallion showed no effect. Until the end of the experiment 2 stallions reached prevaccination testosterone values. Antibody titers varied individually in all 5 stallions and reached peak concentrations within 2 weeks after the third vaccination. Scrotal width was significantly (P<0.05) lower in vaccinated than in control stallions 8 months after first vaccination. Semen quality started to decreased after the second vaccination and improved towards the end of the experiment. In 4 stallions libido was clearly reduced after the second immunization but normalized in 2 animals before the end of the study while 2 stallions continued to show poor libido. From our results we conclude that three immunizations with Equity are well tolerated in the stallion and may effectively suppress testosterone secretion and reduce semen quality as well as sexual behavior. The inhibiting activity of Equity on these parameters is individually different and may last for a minimum of 6 months. PMID- 19128904 TI - Vertebral fracture assessment in Moroccan women: prevalence and risk factors. AB - INTRODUCTION: Vertebral fracture assessment (VFA) is a fast, low-radiation technique which produces images that are of sufficient quality to be used to diagnose the presence of vertebral deformity consistent with fracture. OBJECTIVE: To study prevalence and risk factors of vertebral fractures using VFA in asymptomatic Moroccan women. METHODS: The study cohort consists of a population of 328 consecutive women aged over 50 (mean age, weight and BMI of 65+/-6.5 (50 84) years, 72.0+/-12.8 (42-125) and 29.4+/-5.0 (17.1-45.8)kg/m(2), respectively). Lateral VFA images and scans of the lumbar spine and proximal femur were obtained by two technologists using a GE Healthcare Lunar Prodigy densitometer. Vertebral fractures were defined using a combination of Genant semiquantitative (SQ) approach and morphometry. RESULTS: 68% of vertebrae from T4-L4 and 75% from T8-L4 were adequately visualized on VFA. Vertebral fractures (grades 2 or 3) were detected in 25.6% (84/328) of these women. Thirty-two of women with VFA identified fracture (38.0%) had only a single vertebral fracture, while the other 61.9% had two or more. Fractures were most common in the mid-thoracic spine and at the thoraco-lumbar junction. As would be expected, the prevalence of VFA detected fractures increased with age and as BMD declined. Stepwise regression analysis showed that presence of vertebral fracture was mainly related to the spine osteoporotic status, age older than 65, history of peripheral fracture and more than six parities. CONCLUSION: Vertebral fractures are common in asymptomatic Moroccan women and are related to age, low BMD, history of fracture and multiparity. PMID- 19128903 TI - Relations among menopausal symptoms, sleep disturbance and depressive symptoms in midlife. AB - OBJECTIVES: To investigate the relations among hot flashes, other menopausal symptoms, sleep quality and depressive symptoms in midlife women. METHODS: A large population-based cross-sectional study of 639 women (ages 45-54 years) consisting of a questionnaire including the Center for Epidemiologic Studies Depression (CES-D) Scale, demographics, health behaviors, menstrual history, and menopausal symptoms. RESULTS: After controlling for menopausal status, physical activity level, smoking status and current self-reported health status elevated CES-D score is associated with frequent nocturnal hot flashes, frequent trouble sleeping, experiencing hot flashes, nausea, headaches, weakness, visual problems, vaginal discharge, irritability, muscle stiffness, and incontinence. CONCLUSIONS: The present study found significant links between depressive symptoms and several menopausal symptoms including hot flashes, sleep disturbance, irritability, muscle stiffness, and incontinence after controlling for covariates. These findings suggest that a potential mechanism in which bothersome menopausal symptoms may influence depressed mood during the midlife is through sleep disturbance. PMID- 19128905 TI - Dietary quality indices and human health: a review. AB - Indices are composite tools aiming to measure and quantify a variety of clinical conditions, behaviors, attitudes and beliefs that are difficult to be measured quantitatively and accurately. In this review, the methodology used to develop dietary indices and their relationship with health determinants and outcomes is discussed. In brief, the already proposed indices are adequate tools concerning the evaluation of diet quality, but they have shown moderate predictive ability in relation to chronic diseases and health determinants. The aforementioned weaknesses could be attributed to: inappropriate selection of the components (i.e., number or content of dietary information), selection of small number of cut-off points for each component and/or equal contribution of all index items to the calculation of the total score. Nevertheless, dietary indices are important tools to evaluate not only the diet quality, but also the relationship between dietary habits and several health outcomes. PMID- 19128906 TI - A feasibility study on the use of double-sided adhesive coated stubs for sampling of explosive traces from hands. AB - A study was conducted to assess the applicability of double-sided adhesive coated stubs for sampling traces of TNT, RDX and PETN from suspects' hands. The double stage extraction method developed in the previous study for recovery of NG and 2,4 DNT in gunpowder residues sampled by double-sided adhesive coated stubs, was found to be applicable also for TNT, RDX and PETN. These explosives were analyzed using GC/TEA and IMS. PMID- 19128907 TI - Maternal caloric restriction spares fetal brain polyunsaturated fatty acids in Wistar rats. AB - There is increasing interest in the role of developmental programming; however, the impact on fetal oxidative stress and brain fatty acid levels has been relatively unexplored. Recent reports have shown that caloric restriction regimens in adult animals reduce the occurrence of chronic diseases by reducing the oxidative stress and altering the long chain polyunsaturated fatty acids (LCPUFA). The present study examined whether caloric restriction during pregnancy alters oxidative stress and essential fatty acid metabolism in mother and offspring at birth. Pregnant female rats were fed either a standard chow (C, n=7) or were calorie restricted (CR, n=7) by feeding 60% of the intake of the control. Oxidative stress marker (malondialdehyde) and polyunsaturated fatty acid profiles in brain and liver were analyzed in both dams and offspring. Total weight gain during pregnancy was lower (p<0.01) in the CR group as compared to the control group but did not affect the litter size and weight. Brain malondialdehyde levels were lower (p<0.05) in dams from the CR group. There was no change in brain and liver LCPUFA levels in both male and female offspring in the CR group. Most of the polyunsaturated fatty acids were reduced (p<0.05) in plasma and brain in the CR dams. Caloric restriction during pregnancy did not alter LCPUFA metabolism in the offspring suggesting that during maternal caloric restriction mothers own stores are mobilized to provide docosahexaenoic acid and arachidonic acid for fetal brain development. PMID- 19128908 TI - [Resistance to platelet antiaggregants: the cardiologist's point of view]. AB - The concept of resistance to aspirin and clopidogrel, initially described in the laboratory, has currently been reinforced with recent epidemiological clinical data. One of the elements of particular importance for the cardiologist is the possible participation of this resistance in the process of coronary stent thrombosis, a problem which appeared to be solved early in the 1990s with the introduction of thienopyridines. This complication has however become preoccupying again, particularly when occurring late, notably since the widespread use of biologically active coronary endoprostheses. Nevertheless, the debate continues concerning the usefulness of the biological definition of this concept since we still do not have correctly standardized coherent biological tools that can be used in the clinical setting to detect "resistant" patients. Since there is no real therapeutic strategy which should be applied in the event of resistance, there still is little interest in developing screening methods. But the cardiology community can learn from this concept. We should revisit the principles of revascularization within the framework of the rules of good clinical practice, without speculating about the possible therapeutic finality which might develop should such and such a phenomenon occur. PMID- 19128909 TI - Prevalence of nephrogenic systemic fibrosis in renal insufficiency patients: results of the FINEST study. AB - PURPOSE: Nephrogenic systemic fibrosis (NSF) is characterized by widespread tissue fibrosis, mainly affecting the skin. Gadolinium chelates have been implicated in the onset of NSF in patients with renal impairment (RI). The FINEST study (FIbrose Nephrogenique SysTemique) was designed to determine the prevalence of NSF after magnetic resonance imaging (MRI) in French RI patients. MATERIALS AND METHODS: We studied all patients with RI who had at least one MRI examination during a one-year period, with or without gadolinium chelate administration. Data were collected retrospectively from 9 Nephrology Departments in France, and included sex, age, renal function, type of gadolinium administered, and subsequent cutaneous disorders. If a patient presented a cutaneous disorder, a skin biopsy was performed to confirm the diagnostic. RESULTS: The 308 eligible patients had a mean age of 59.9 years, 59% were men, and 54% had stage 5 RI. 75% of those 308 patients received a Gadolinium chelate. Among those patients who received a gadolinium chelate, 76% received gadoterate, 20% gadopentetate, 3% gadodiamide and 1% gadobenate. No cutaneous disorders were recorded after MRI. CONCLUSION: These results confirm that NSF is a rare disease. Based on a reported frequency, approximately 3.5% in patients with glomerular filtration rate <30ml/min/1.73m(2)), some cases should have been observed in our study which included 308 patients. Most patients received gadoterate, a macrocyclic gadolinium chelate for which no case of NSF has been observed worldwide. This suggests that more stable macrocyclic agents may be less likely to induce NSF. PMID- 19128910 TI - Mapping the human brain white matter tracts relative to cortical and deep gray matter using diffusion tensor imaging at high spatial resolution. AB - The mapping of the human brain white matter fiber networks relative to deep subcortical and cortical gray matter requires high spatial resolution which is challenged by the low signal-to-noise ratio. The purpose of this short report was to introduce a whole brain high spatial resolution diffusion tensor imaging (DTI) protocol that enabled for the first time the mapping of corticopontocerebellar, frontostriatal and thalamofrontal fiber pathways in addition to other limbic, commissural, association and projection white matter pathways relative to the segmented deep gray (e.g., caudate nuclei) and the cortical lobes. Our DTI acquisition protocol and analysis strategy provide important template for brain behavior research and for teaching brain mapping and are clinically affordable for patient comfort. PMID- 19128911 TI - Quantification of the HIV-integrase inhibitor raltegravir and detection of its main metabolite in human plasma, dried blood spots and peripheral blood mononuclear cell lysate by means of high-performance liquid chromatography tandem mass spectrometry. AB - For the quantification of the HIV-integrase inhibitor raltegravir in human plasma, dried blood spots and peripheral blood mononuclear cell (PBMC) lysate, an assay was developed and validated, using liquid chromatography coupled with tandem mass spectrometry. The assay also allowed detection, but no quantification due to absence of reference substance, of the main metabolite, raltegravir glucuronide. Raltegravir was extracted from plasma by means of protein precipitation with a mixture of methanol and acetonitrile using only 50microL plasma. Extraction from dried blood spots was performed with a simple one-step extraction with a mixture of methanol, acetonitrile and 0.2M zincsulphate in water (1:1:2, v/v/v) and extraction from cell lysate was performed in 50% methanol in water. Chromatographic separation was performed on a reversed phase C18 column (150mmx2.0mm, particle size 5microm) with a quick stepwise gradient using an acetate buffer (pH 5) and methanol, at a flow rate of 0.25mL/min. The analytical run time was 10min. The triple quadrupole mass spectrometer was operated in the positive ion-mode and multiple reaction monitoring was used for drug quantification. The method was validated over a range of 50-10,000ng/mL in plasma and dried blood spots and a range of 1-500ng/mL in PBMC lysate. Dibenzepine was used as the internal standard. The method was proven to be specific, accurate, precise and robust. Accuracies ranged from 104% to 105% in plasma, from 93% to 105% in dried blood spots and from 82% to 113% in PBMC lysate. Precision over the complete concentration range was less than 6%, 11% and 13% in plasma, dried blood spots and PBMC lysate, respectively. The method is now applied for therapeutic drug monitoring and pharmacological research in HIV infected patients treated with raltegravir. PMID- 19128912 TI - Oligonucleotide chip assay for quantification of gamma ray-induced single strand breaks. AB - An oligonucleotide chip assay was designed for direct quantification of single strand breaks (SSBs) induced by gamma-ray irradiation. The oligonucleotides used were 20-mers, which were short enough to produce only a single strand break within a single oligonucleotide. The two ends of the oligonucleotides were labeled with fluorescein and biotin, respectively. The biotinylated ends of the oligonucleotides were immobilized on a silicon wafer chip treated with (3 aminopropyl)triethoxysilane (APTES), glutaraldehyde, and avidin. The DNA fragments cleaved by gamma-ray irradiation were detected by a laser-induced fluorescence (LIF) detection system. The gamma-ray-induced SSBs were quantified using a calibration curve (fluorescence intensity versus gamma-ray dose) without the need for complicated mathematical calculation based on gel-based separation. The experimentally determined gamma-ray-induced SSBs yield was almost equal to the theoretical value derived from gel electrophoresis of plasmid DNAs and DNA surface coverage. PMID- 19128913 TI - Genetic diversity of genes encoding OKP and LEN beta-lactamases produced by clinical Klebsiella pneumoniae strains in Portugal. AB - Of the 308 clinical Klebsiella pneumoniae strains collected in 21 Portuguese health institutions, 11 encoded for LEN and 9 for OKP enzymes; of these, 15 were new enzymes. Ninety-one percent of LEN and all OKP producer strains were resistant to amoxicillin. We demonstrate that these beta-lactamase were highly diverse. PMID- 19128914 TI - Spontaneous spinal epidural hematoma: a case report and literature review. AB - BACKGROUND: Spontaneous spinal epidural hematoma (SSEH) is a rare cause of spinal cord compression that requires emergency investigation and treatment. Prompt diagnosis is essential to prevent morbidity and mortality. OBJECTIVES: Our goal was to report a case of SSEH, and to review the literature on the topic, looking particularly at the factors influencing post-operative outcome, and the symptoms and signs that would lead one to consider this rare diagnosis. CASE REPORT: A 36 year-old man presented to the Emergency Department with paraplegia and a sensory level at T4. There was no history of trauma and he was not taking any medications. Magnetic resonance imaging revealed a large non-enhancing posterior epidural mass lesion between C7 and upper T4, causing severe cord compression at T1-T3. Post contrast scans revealed no rim enhancement. Blood investigations were unremarkable, with an international normalized ratio of 1.1. He underwent urgent decompression laminectomy within 12 h of symptom onset. Intraoperatively, thick clotted blood and prominent epidural vessels were seen. Histology revealed engorged vessels with hemorrhage and clot in the extravascular tissues. Post operatively, the impaired neurological status remained unchanged. CONCLUSION: SSEH is a rare cause of spinal cord compression that requires prompt diagnosis and surgical intervention to prevent morbidity. Neurological outcome after surgical decompression depends on the severity of preoperative neurological deficits as well as the time between symptom onset and surgery. PMID- 19128915 TI - Deep venous thromboembolism in a triathlete. AB - BACKGROUND: Demanding athletic events can contribute multiple risk factors to the development of deep venous thrombosis (DVT) in athletes. OBJECTIVES: To discuss risk factors for the development of DVT in athletes participating in athletic events. CASE REPORT: We present the case of a young female athlete who had a DVT, complicated by pulmonary embolism, after participating in a half-Ironman triathlon. CONCLUSION: Our patient developed a DVT complicated by pulmonary embolism as a result of many factors relating to participation in a triathlon. Demanding athletic events can contribute different risk factors to the development of DVT in athletes. The diagnosis of DVT should be considered in any athlete presenting with leg pain, especially after a strenuous athletic event. PMID- 19128917 TI - Penile fracture in a patient presenting with groin pain. PMID- 19128918 TI - Sensitivity of screening tests and other questions. PMID- 19128919 TI - Trends in boarding of admitted patients in US Emergency Departments 2003-2005. AB - BACKGROUND: Boarding of admitted patients in the Emergency Department (ED) is common and is associated with poor patient outcomes. OBJECTIVES: We sought to estimate the magnitude of and trends for ED boarding in the US. METHODS: We used the 2003-2005 National Hospital Ambulatory Medical Care Survey to estimate the time patients spent boarding in EDs in the US. We used fixed and imputed times required to evaluate, treat, and decide to admit each patient using the number of medications and diagnostic tests received. We calculated the absolute and relative patient-care hours spent boarding in US EDs over the 3-year period. RESULTS: Total patient-hours spent in US EDs increased from 209 million to 217 million between 2003 and 2005. Overall admission rates decreased between 2003 and 2005 (13.9% in 2003, 12.3% in 2005), whereas intensive care unit admission rates increased (1.3% in 2003, 2.0% in 2005). Mean ED length of stay decreased (5.4 h in 2003, 4.6 h in 2005). The proportion of patient-hours accounted for by ED boarding decreased over the study period (11.3-17.1% in 2003, 5.9-15.3% in 2004, and 2.8-12.0% 2005). CONCLUSIONS: Boarding of admitted patients in the ED accounts for a substantial portion of ED patient-care hours. Overall boarding time decreased over the 3 years. PMID- 19128920 TI - A significant cervical spine fracture: think of the airway. AB - BACKGROUND: Airway compromise is a potential complication of significant cervical spine injury. OBJECTIVES: To alert emergency physicians to be aware of possible airway collapse after serious cervical spine injury. CASE REPORT: We report a case of an 87-year-old man who presented to the emergency department with an unstable cervical spine fracture after a fall. He subsequently developed complete upper airway obstruction from prevertebral soft tissue swelling, requiring a cricothyrotomy after a failed intubation attempt. CONCLUSION: Patients with significant blunt cervical spine trauma can be at high risk for upper airway compromise. PMID- 19128922 TI - Non-invasive pulse CO-oximetery: what is measured? PMID- 19128923 TI - Serum C-reactive protein concentration and the prognosis of ductal adenocarcinoma of the head of pancreas. AB - BACKGROUND: The prognostic role of serum C-reactive protein in pancreatic cancer has received increasing attention; however the confounding effects of biliary obstruction have not been addressed in previous studies. We sought to determine the prognostic importance of serum CRP prior to biliary intervention in the prognosis of pancreatic adenocarcinoma. METHODS: A retrospective case note review of patients diagnosed with pancreatic cancer between 2001 and 2006. Clinical, radiological and biochemical criteria were correlated with overall survival. Patients were divided into: Group 1 who underwent potentially curative resection, and Group 2 with advanced unresectable disease managed non-surgically. RESULTS: In total, 199 patients were included (58 resected). The proportion of patients with biliary obstruction was equal in both groups. Serum CRP and serum bilirubin concentration at presentation were significantly higher among patients in Group 2 compared to Group 1 (P values). On multivariate analysis, advancing age (P=0.012) and raised serum CRP concentration were independently associated with overall survival only in Group 2 patients (P=0.027, 95% CI 0.31-0.93). This association was independent of biliary tract obstruction. CONCLUSION: Raised serum C-reactive protein concentration at the time of presentation of advanced pancreatic cancer carries a poor prognosis independent of biliary tract obstruction. PMID- 19128924 TI - [Severe hyponatraemia secondary to cure of cyclophosphamide (about three cases)]. PMID- 19128925 TI - [Tuberculosis risk among patients with systemic diseases]. AB - The incidence of tuberculosis among patients with systemic rheumatic diseases is much higher than in the general population (the risk is multiplied by 5 to 15 in patients with systemic lupus erythematosus). Reactivation of a latent tuberculosis is frequent, as assessed by the short delay of occurrence after a systemic rheumatic disease has been diagnosed. Besides immunosuppression induced by the underlying disease, the role of glucocorticoids and of immunosuppressive therapy including biotherapies using TNF antagonists must be underlined. Tuberculosis in such patients frequently presents as extrapulmonary or disseminated disease. A screening of tuberculosis is recommended before anti-TNF therapy, and includes previous history questioning, chest X ray, tuberculin skin test with 5 international units of tuberculin. Immunological methods of tuberculosis detection are under evaluation in these patients. If a latent tuberculosis infection is diagnosed, a specific tuberculosis chemoprophylaxis, started at least 3 weeks before initiation of TNF antagonists, has allowed to reduce the occurrence of anti-TNF-associated tuberculosis in patients living in Europe and North America. The screening strategies for tuberculosis should probably be extended in all patients with systemic rheumatic diseases receiving glucocorticoids and/or immunosuppressive therapy. PMID- 19128926 TI - The age-adjusted Charlson Comorbidity Index as an outcome predictor of patients with acute mesenteric ischemia. AB - Acute mesenteric ischemia (AMI) is a catastrophic surgical condition, especially in older patients with multiple comorbidities. The aim of this study was to evaluate the impact of comorbidity on perioperative mortality and overall survival in patients surgically treated for AMI. A series of 186 consecutive patients (106 men and 80 women) who underwent surgery because of AMI in a university tertiary care center between 1990 and 2006 were retrospectively studied. The Charlson Comorbidity Index (CCI) score, unadjusted and adjusted by age, was preoperatively calculated in each patient. Perioperative mortality and overall survival were also recorded. The association between unadjusted and adjusted by age CCI and perioperative mortality and overall survival were analyzed. The mean age of the studied population was 72.1 years (SE +/-13.7 years). Hospital mortality was 64.5%. One-year, 3-year, and 5-year overall estimated survival by the Kaplan-Meier method after surgery for AMI was 26%, 23% and 21%, respectively. Perioperative mortality was not related to the unadjusted preoperative CCI (p = 0.093). Nevertheless, a statistically significant association was found between mortality and preoperative adjusted CCI (p = 0.007). Likewise, CCI unadjusted was almost related to overall survival (p = 0.055), but the values of the categorized CCI adjusted by age showed a statistically significant difference in overall survival (p = 0.012). In multivariate analysis, CCI adjusted by age remained independent prognostic factor of mortality. Comorbidity adjusted by age may play a role as a predictive factor for perioperative mortality and long-term survival in patients operated on for AMI. PMID- 19128927 TI - Systematic review of endovenous laser therapy versus surgery for the treatment of saphenous varicose veins. AB - This systematic review compares the safety and efficacy of endovenous laser therapy (ELT) and surgery involving saphenous ligation and stripping as treatments for varicose veins. Systematic searches of medical bibliographic databases, the Internet and lists of references were conducted in August 2007 and April 2008 to identify relevant primary studies. Inclusion of papers was resolved through application of a predetermined protocol. Information on the safety and effectiveness of ELT and surgery was analyzed. Fifty-nine studies were included, with seven studies directly comparing ELT with surgery. Serious adverse events after ELT or surgery were rare. While occurrence rates of some minor adverse events appeared higher after ELT in collated data, comparative studies commonly favored ELT over surgery. Few differences were apparent between treatments with respect to clinical effectiveness outcomes, although long-term follow-up was lacking. Nonclinical effectiveness outcomes generally favored ELT over surgery in the first 2 months after treatment. ELT appears to be at least as safe as surgery. While ELT offers short-term benefits and appears to be as clinically effective as surgery up to 12 months after treatment, clinical trials with a minimum of 3 years of follow-up are required to establish the enduring effectiveness of ELT. PMID- 19128928 TI - What is the evidence on efficacy of spinal cord stimulation in (subgroups of) patients with critical limb ischemia? AB - The use of spinal cord stimulation (SCS) has been advocated for the management of ischemic pain and the prevention of amputations in patients with inoperable critical limb ischemia (CLI), although data on benefit are conflicting. Several reports described apparently differential treatment effects in subgroups. The purpose of this study was to analyze the data on the efficacy of SCS and to clarify preselection issues. Five randomized trials have been performed with a total number of 332 patients. Primary outcome measures were mortality and limb survival. In the largest multicenter randomized trial (n = 120), which compared SCS treatment and best medical treatment alone in patients with inoperable CLI, we determined the incidence of amputation and its relation to various predefined risk factors. We used Kaplan-Meier and Cox regression analyses to quantify prognostic effects and differential treatment effects. Meta-analysis yielded a relative risk for amputation of 0.79 and a risk difference of -0.07 (p = 0.15). The risk factor analysis clearly showed that patients with ischemic skin lesions (ulcerations or gangrene) had a worse prognosis (i.e., higher risk of amputation) (relative risk 2.30, p = 0.01). We did not observe significant interactions between this prognostic factor (or any other) and the effect of SCS. The analysis did not indicate a subgroup of patients who might specifically be helped by SCS. Meta-analysis including all randomized data shows insufficient evidence for higher efficacy of SCS treatment compared with best medical treatment alone. Although some factors provide prognostic information as to the risk of amputation in patients with CLI, there are no data supporting a more favorable treatment effect in any group. PMID- 19128929 TI - Management of median arcuate ligament syndrome: a new paradigm. AB - Median arcuate ligament (MAL) syndrome is an anatomic and clinical entity characterized by extrinsic compression on the celiac axis, which leads to postprandial epigastric pain, vomiting, and weight loss. Although characterized a few decades ago, the existence of this syndrome is still challenged by several authors. We reviewed the management of MAL syndrome, with special emphasis on the minimally invasive approaches. We also report the first case of successful combination of minimally invasive surgery and endovascular therapy in the treatment of this syndrome. A PubMed search was carried out to identify articles in English from 1963 to 2008 using the keywords "median arcuate ligament syndrome" and "celiac artery compression syndrome." Additional articles were identified by a manual search of the references from the key articles. All clinical and experimental studies that contained material applicable to the topic were considered. Classic treatment of the condition is represented by open MAL release. However, permanent changes in the celiac artery wall lead to poor long term outcomes, and associated complex vascular procedures have been employed. Laparoscopic treatment of MAL syndrome was reported in five cases. All patients had resolution of symptoms, but long-term follow-up is not available. Laparoscopic release of arcuate ligament syndrome with intraoperative duplex ultrasound may be used in patients with symptoms suggestive of the diagnosis. In patients with persistent celiac flow abnormalities noted on duplex ultrasound or postoperative imaging, celiac angioplasty and stenting are advocated. If this option is not available or does not relieve symptoms, vascular reconstruction should be employed. PMID- 19128930 TI - The role of oxidative stress and antioxidant defenses in Buerger disease and atherosclerotic peripheral arterial occlusive disease. AB - The aim of this study was to determine the status and the role of oxidative stress and antioxidant defenses in patients with Buerger disease and atherosclerotic peripheral arterial occlusive disease (PAOD). Seventy-three subjects resembling each other in general characteristics were involved in the study: 21 with lower extremity PAOD (mean age 53.05 +/- 10.8 years, 17 men and four women), 22 with Buerger disease (mean age 38.59 +/- 6.4 years, 19 men and three women), and 30 healthy volunteers (mean age 38.59 +/- 6.4 years, 22 men and eight women). We measured the levels of plasma malondialdehyde (MDA), paraoxonase (PON1), protein carbonyls, arylesterase, nitric oxide (NO), serum oxidized low density lipoprotein (ox-LDL) and MDA, glutathione (GSH), glutathione reductase (GSH-red), glutathione peroxidase (GSH-px), superoxide dismutase (SOD), and catalase (CAT) in erythrocytes. Plasma protein carbonyls, serum ox-LDL, and plasma and erythrocyte MDA were significantly high in the Buerger disease group compared to the PAOD and control groups (p < 0.001). Plasma PON1 levels and GSH and GSH-px levels in erythrocytes in the Buerger disease group were significantly low compared to the PAOD and control groups (p < 0.001). GSH-red, SOD, and CAT levels in erythrocytes in the Buerger disease group were significantly lowcompared to the PAOD group (p < 0.01, p < 0.001, and p < 0.001, respectively). NO levels were significantly lower in the PAOD group compared to the control group (p < 0.05). The balance between oxidative stress and antioxidant capacity is more seriously impaired in Buerger disease than PAOD. PMID- 19128931 TI - General versus vascular surgeon: impact of a vascular fellowship on clinical practice, surgical case load, and lifestyle. AB - An applicant shortage for vascular surgery (VS) residencies exists despite an increase in available training positions created to meet the growing demands for vascular surgeons. After 3 years of practice as an American Board of Surgery (ABS)-certified/board-eligible general surgeon, the author of this study attended an accredited 1-year VS training fellowship and received an ABS certificate of Added Qualifications in VS. The purpose of this review was to investigate the implications completing a vascular fellowship has had on VS procedure patterns, vascular procedure competency, clinical practice, career, and lifestyle with the aim of attracting trainees to the field of VS. The author's operative logs were reviewed retrospectively to summarize vascular procedures performed before and after the vascular fellowship. Statistical analysis was performed comparing the types and volume of vascular procedures before and after the vascular fellowship. Changes in professional career and personal life also were examined. The author performed 401 vascular procedures during 2.8 years as a general surgeon. In the first 3.4 years after the vascular fellowship, vascular procedure volume increased to 1563. The mean number of vascular procedures performed per year increased from 143.2 as a general surgeon to 459.7 as a vascular surgeon. The three major differences in vascular procedures occurring after the vascular fellowship were (1) a threefold increase in the number of vascular procedures performed, (2) a shift from major open to venous and endovascular procedures, and (3) an increase in case complexity. Specializing in VS also has resulted in increased career opportunities, more career satisfaction, a direct financial benefit, and more flexibility for lifestyle and family. Because of these positive changes, the author encourages medical students and residents interested in VS to explore the specialty early, seek vascular surgeons to serve as mentors, and enter one of the new VS pathways as early as possible in their career. PMID- 19128932 TI - Complications during the waiting period for carotid endarterectomy. AB - We conducted a retrospective analysis to determine the number of cerebrovascular complications occurring in the period between initial presentation and operation in patients undergoing carotid endarterectomy. The notes of patients undergoing carotid endarterectomy during a 2-year period were used to quantify delays between initial presentation and surgery and complications occurring in this time period. A group of 49 patients was analyzed. The median time period between referral following initial cerebrovascular event and carotid endarterectomy was 62 days. During this time period 12 (24.5%) ischemic complications occurred including six transient ischemic attacks. Delays occurred at a variety of levels but included a median of 36 days before patients seen in the stroke clinic were referred to a surgeon and a median of 23 days before operation once patients had seen a surgeon. Significant delays still exist between initial presentation and carotid endarterectomy and can lead to further ischemic episodes prior to surgery. PMID- 19128933 TI - Carotid stenting using tapered and nontapered stents: associated neurological complications and restenosis rates. AB - Self-expanding stent design systems for carotid artery stenting (CAS) have morphed from nontapered (NTS) to tapered (TS); however, the impact of this change is unknown. We reviewed the outcomes of CAS with these two broad categories of stents in a single-center retrospective review of 308 CAS procedures from May 2001 to July 2007. Nitinol self-expanding TS or NTS coupled with cerebral embolic protection devices were used to treat extracranial carotid occlusive disease. Data analysis included demographics, procedural records, duplex exams, and conventional arteriography. Mean follow-up was 18 months (range 1-69). Restenosis was defined as >or=80% in-stent carotid artery stenosis by angiography. The mean age of the entire cohort was 71.3 years (75% men, 25% women). Of the 308 cases, 233 were de novo lesions and 75 had a prior ipsilateral carotid endarterectomy (n = 44) or external beam radiation exposure (n = 31). Preprocedure neurological symptoms were present in 30% of patients. TS were used in 156 procedures and NTS in 152 procedures. The 30-day ipsilateral stroke and death rates were 1.3% and 0.3%, respectively. An additional three (1.0%) posterior circulation strokes occurred. There was no statistically significant difference in the 30-day total stroke rates between TS (3.2%, n = 5) and NTS (1.3%, n = 2) (p = 0.5). At midterm follow-up, restenosis or asymptomatic occlusion was detected in eight cases (2.6%). All occurred in arteries treated with NTS, and this was statistically different when compared to arteries treated with TS (p = 0.03). Furthermore, a post-hoc subgroup analysis revealed significant correlation (chi(2) = 0.02) for restenosis in "hostile necks" when separated by TS vs. NTS. Early CAS outcomes between TS and NTS are comparable. In contrast, self-expanding nitinol TS may have a lower incidence of significant restenosis or asymptomatic occlusion when compared to NTS. PMID- 19128934 TI - Impact of chronic kidney disease on outcomes of superficial femoral artery endoluminal interventions. AB - While aggressive endoluminal therapy for superficial femoral artery (SFA) occlusive disease is commonplace, the implications of chronic kidney disease (CKD) on long-term outcomes in this population are unclear. We examined the consequences of endovascular treatment of the SFA in patients with and without varying stages of CKD. A database of patients undergoing endovascular treatment of the SFA between 1986 and 2007 was queried, and two groups were defined: estimated glomerular filtration rate (eGFR) 60 mL/min/1.73 cm(2). Intention-to-treat analysis was performed. Results were standardized to TransAtlantic Inter-Society Consensus (TASC-II) and Society for Vascular Surgery criteria. Kaplan-Meier analyses were performed to assess time-dependent outcomes. Factor analyses were performed using a Cox proportional hazard model for time dependent variables. Data are presented as mean +/- standard deviation where appropriate. There were 525 limbs in 535 patients (68% male, average age 66 +/- 14 years) that underwent endovascular treatment for claudication or chronic critical limb ischemia (51%). Patients with eGFR 60. In patients with critical limb ischemia, there was no difference in patency between those with eGFR 60. Limb salvage was worse in patients with eGFR 60. With respect to limb salvage, six factors were significantly associated with a reduction in rates: presence of tissue loss at presentation (relative risk [RR] = 6.45, p = 0.003), 0 or 1 vessel tibial runoff (RR = 2.56, p < 0.01), progression of distal disease noted in follow-up (RR = 4.62, p < 0.01), embolization at the initial intervention (RR = 2.70, p < 0.05), diabetes mellitus (RR = 3.71, p < 0.01), and a history of congestive heart disease (RR = 2.42, p < 0.01). Notable factors that were not significantly associated included lesion calcification (p = 0.64), TASC C or D lesion categorization (p = 0.99), acute occlusion at initial intervention (p = 0.40), and adjuvant stenting (p = 0.67). CKD does not impact the patency of SFA interventions. Limb salvage in patients with critical ischemia is significantly worse when the eGFR is 20%. Trials that were eventually published had larger mean population sizes (P-value, 0.02). Of the reported sponsors, 87.3% were drug companies. Methodological quality and publication bias are mentioned in half of the reviews and explored in a third. Quality ratings did not have consequences for pooling, because 82.8% was included in the forest plots. CONCLUSIONS: A minority of Cochrane reviews include "unpublished trials" and many of these are eventually published. Truly unpublished studies have poor or unclear methodological quality. Therefore, it may be better to invest in regular updating of reviews, rather than in extensive searching for unpublished data. PMID- 19128940 TI - Non-Cochrane vs. Cochrane reviews were twice as likely to have positive conclusion statements: cross-sectional study. AB - OBJECTIVES: To determine which factors predict favorable results and positive conclusions in systematic reviews (SRs) and to assess the level of agreement between SR results and conclusions. STUDY DESIGN AND SETTING: A sample of 296 English SRs indexed in MEDLINE (November, 2004) was obtained. Two investigators independently categorized SR characteristics, results, and conclusions. Descriptive analyses and logistic regression predicting favorable results (nonstatistically significant and statistically significant positive) and positive conclusions were conducted. The level of concordance between results and conclusions was assessed using a weighted-kappa statistic. RESULTS: Overall, 36.5% of the SRs had favorable results, increasing to 57.7% for Cochrane and 64.3% for non-Cochrane reviews with a meta-analysis of the primary outcome. Non Cochrane reviews with a meta-analysis of the primary outcome were twice as likely to have positive conclusions as Cochrane reviews with such an analysis (P value<0.05). The weighted kappa for agreement between SR results and conclusions was 0.55. It was lower for Cochrane (0.41) vs. non-Cochrane (0.67) reviews. CONCLUSION: SRs including a meta-analysis of the primary outcome may be affected by indirect publication bias in our sample. Differences between the results and conclusions of Cochrane and non-Cochrane reviews were apparent. Further research on publication-related issues of SRs is warranted. PMID- 19128941 TI - Citations to trials of nicotine replacement therapy were biased toward positive results and high-impact-factor journals. AB - OBJECTIVE: To study variations in the number of times trials of nicotine replacement therapy (NRT) were cited, and which characteristics of trials predicted the number of citations and the impact factors of journals in which articles were published. STUDY DESIGN AND SETTING: We used all 105 randomized controlled trials in the Cochrane review of NRT for smoking cessation. We obtained impact factors from the Journal Citation Reports and the number of citations from ISI Web of Knowledge and Google Scholar. RESULTS: Trials were cited from 0 to 632 times (median 23 times). Trials were cited more often when results were statistically significant than when they were not (median=41 vs. 17 times, P<0.001), and when impact factors were higher (10.2 more citations per impact factor point, P<0.001). Patch trials were cited more often than gum trials (median=29 vs. 17 times, P=0.001), and trials funded by the pharmaceutical industry were cited more often than other trials (median=28 vs. 16.5 times, P=0.001). Trials with statistically significant results were published in journals with higher impact factors than trials with nonsignificant results (median impact factor=2.80 vs. 1.81, P=0.011). CONCLUSION: Citations were biased toward trials with positive results and toward trials published in high-impact factor journals. PMID- 19128942 TI - Clinical survey of neurosensory side-effects of mandibular parasymphyseal bone harvesting. AB - The aim of the present survey was to assess neurosensory disturbances and/or tooth-pulp sensitivity losses after mandibular parasymphyseal bone-harvesting procedures. Twenty-eight harvesting areas in 16 patients were surveyed. Mucosal and skin sensitivity of the chin/lower lip, divided into four regions, were determined via Pointed-Blunt and Two-Point-Discrimination Tests. Pulp sensitivity of the mandibular teeth from the left second bicuspid to the right second bicuspid was tested by cold vitality preoperatively and 12 months postoperatively. Teeth were grouped according to sensitivity alterations and distance from the harvesting defects, as measured on CT scans, and statistically significant differences sought. At 12 months, 29% of preoperatively vital cuspids overlying the harvesting defects revealed pulp-sensitivity losses; no patient reported anaesthesia or analgesia; hypoaesthesia was present in 4% (8 sites; 2 patients), hypoalgesia was present in 3% (5 sites; 2 patients) and Two-Point Discrimination Tests yielded pathologic responses in 5% of tested areas (10 sites; 4 patients). Teeth with and without pulp sensitivity changes were statistically indistinguishable regarding distances between root apices or mental foramen and the harvesting defect. The loss of pulp sensitivity in any tooth cannot be predicted simply on the basis of the distance between its apex and the harvesting osteotomy line. PMID- 19128943 TI - Maxillo-mandibular counter-clockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses: part III--pain and dysfunction outcomes. AB - 47 end-stage TMJ patients with high occlusal plane angulation, treated with TMJ custom-fitted total joint prostheses and simultaneous maxillo-mandibular counter clockwise rotation were evaluated for pain and dysfunction presurgery (T1) and at the longest follow-up (T2). Patients subjectively rated their facial pain/headache, TMJ pain, jaw function, diet and disability. Objective functional changes were determined by measuring maximum interincisal opening (MIO) and laterotrusive movements. Patients were divided according to the number of previous failed TMJ surgeries: Group 1 (0-1), Group 2 (2 or more). Significant subjective pain and dysfunction improvements (37-52%) were observed (90% of cases. Among MATR twins with a verified history of epilepsy, 21.5% reported other seizures but not epilepsy and 18.5% of verified Norwegian epilepsy cases reported no history of epilepsy themselves and were identified only through their co-twin. The results of this study indicate that the accuracy of self-reported epilepsy and febrile seizures among those who provided information on health history was high across all populations. However, the relatively large percentage of twins with a verified diagnosis who did not acknowledge epilepsy suggests that the frequency of epilepsy may be under estimated in self-reported samples. PMID- 19128945 TI - Meta-analysis of diffusion tensor imaging studies in schizophrenia. AB - The objective of the study was to identify whether there are consistent regional white matter changes in schizophrenia. A systematic search was conducted for voxel-based diffusion tensor imaging fractional anisotropy studies of patients with schizophrenia (or related disorders) in relation to comparison groups. The authors carried out meta-analysis of the co-ordinates of fractional anisotropy differences. For the meta-analysis they used the Activation Likelihood Estimation (ALE) method hybridized with the rank approach used in Genome Scan Meta-Analysis (GSMA). This system detects three-dimensional conjunctions of co-ordinates from multiple studies and permits the weighting of studies in relation to sample size. Fifteen articles were identified for inclusion in the meta-analysis, including a total of 407 patients with schizophrenia and 383 comparison subjects. The studies reported fractional anisotropy reductions at 112 co-ordinates in schizophrenia and no fractional anisotropy increases. Over all studies, significant reductions were present in two regions: the left frontal deep white matter and the left temporal deep white matter. The first region, in the left frontal lobe, is traversed by white matter tracts interconnecting the frontal lobe, thalamus and cingulate gyrus. The second region, in the temporal lobe, is traversed by white matter tracts interconnecting the frontal lobe, insula, hippocampus-amygdala, temporal and occipital lobe. This suggests that two networks of white matter tracts may be affected in schizophrenia, with the potential for 'disconnection' of the gray matter regions which they link. PMID- 19128946 TI - Efficient analysis of (51)V solid-state MAS NMR spectra using genetic algorithms. AB - A program for iterative fitting procedures to determine the NMR parameters from (51)V solid-state MAS NMR spectra was developed. It contains options to use genetic algorithms and downhill-simplex optimizing procedures to extract the optimal parameter sets, which describe our spectra. As computational kernel the SIMPSON program is employed. Other kernels like SPINEVOLUTION are easily incorporable. The algorithms are checked for their suitability for the present optimization problem and optimal simulation conditions are determined, with the focus on minimal processing time. The procedure leads to a very good agreement between experimental and simulated spectra in a passable period of time. First results for spectra of model compounds for the active site of vanadium haloperoxidases are presented. PMID- 19128947 TI - [Muscular complications of hemophilia]. AB - Haemophilia causes principally bleedings in muscles and joints. Therapy is purely palliative, based on injections of antihemophilic factor. Muscular hematomas represent the most precocious complication and generally happen after a trauma. Though they scarcely threaten the life of the patients, they often may generate an important functional disability if not treated quickly, and vigorously. Treatment should associate injections of the defective clotting factor to physiotherapy and contractions of the involved muscle. When hematomas involve lower limbs, standing up and walking should not be authorized before the end of the flessum or recovering of the statu quo ante. PMID- 19128948 TI - Carbofuran-induced toxicity in rats: protective role of vitamin C. AB - Erythrocytes are prone to oxidative stress due to the presence of hemoglobin and polyunsaturated fatty acids. Oxidative stress (OS) is associated with increased osmotic fragility (OF) of erythrocytes. Organophosphate and organocarbamate pesticides are known to cause OS in erythrocytes. We have investigated the effect of a single sub-acute dose of carbofuran (CF), an organocarbamate pesticide and ameliorating role of vitamin C on OF and OS in erythrocytes of Wistar rats. OF and OS were assessed by determining membrane stability in terms of erythrocyte OF and the activities of free radicals scavenging enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST). We observed a significant alteration in the mean erythrocyte fragility (MEF) at relatively higher NaCl concentration (0.67%) as compared to MEF at 0.55%, 0.58% and 0.56% of NaCl in control, vitamin C- and vitamin C + CF-treated groups, respectively. The activities of CAT and SOD were observed to be elevated by 74.35% and 85.56%, respectively, with significance level of p < or = 0.001, whereas GST activity got significantly (p < or = 0.001) diminished by 46.30% in the erythrocytes of CF treated rats. Vitamin C treatment exhibited marked (p < or = 0.05) prevention of carbofuran-induced oxidative stress as well as erythrocyte osmotic fragility in the Wistar rats. These results suggest that CF treatment induces OF and OS in the erythrocytes of rats, and pretreatment with vitamin C can mitigate these toxic effects. PMID- 19128949 TI - Orientation of the rotator cuff insertion facets on the humerus: comparison between individuals with intact and torn rotator cuffs. AB - The purpose of this work has been to compare the orientation of the rotator cuff insertions on the humerus between individuals with intact and torn rotator cuffs, respectively, to look for anatomical factors involved in the pathogenesis of rotator cuff tears. A total of 60 patients evaluated by arthro-computed tomography (CT) were divided into an intact rotator cuff group (31 patients) and a torn rotator cuff group (29 patients). The orientation angles of the lesser humeral tubercle facet and of the superior, middle and inferior facets of the greater humeral tubercle were measured in the sagittal plane between the horizontal line (defined as perpendicular to the diaphysis long axis) and the line tangent to the facet. The orientation angles of the superior and middle facets of the greater humeral tubercle were measured in the coronal plane. The comparison between the two groups showed a significant difference (p=0.009) concerning the orientation of the middle facet of the greater tubercle in the sagittal plane, with a mean angle of 36.0 degrees (S.D., 6.2 degrees) in the intact cuff group versus 31.1 degrees (S.D.,7.2 degrees) in the torn cuff group. There were no other significant differences in regard to the orientation of the facets in the sagittal and coronal planes. A decrease in the dorsal orientation of the middle facet of the greater tubercle, as we observed in the torn rotator cuff group, may reduce the inferior translatory forces generated by the infraspinatus thereby facilitating impingement of the subacromial structures. These additional anatomical data might augment the body of knowledge on rotator cuff pathology. PMID- 19128950 TI - The in vitro effects of cigarette smoke on fatty acid metabolism are partially counteracted by simvastatin. AB - BACKGROUND: Statins enhance the synthesis of long-chain polyunsaturated fatty acids (LC-PUFAs) from their precursors both in vitro and in vivo. In particular, an increased conversion of linoleic acid (LA) and of alpha-linolenic acid to their derivatives is observed in cultured cells. On the contrary, cigarette smoke (CS) negatively and dose-dependently affects the LC-PUFA production. AIM: To evaluate the effects of CS alone or with simvastatin, on [1-(14)C] LA metabolism in THP-1 cells. RESULTS: CS inhibits LA conversion; after co-incubation, simvastatin nullifies the effects of CS, maintaining LA conversion comparable to controls. However, at the highest CS concentration, simvastatin is unable to counteract the effects of CS. Changes of LA conversion reflect the modulation of desaturase activities by simvastatin and CS. CONCLUSION: CS decreases PUFA conversion and its effects are modulated by the opposite effect of statins. It can be speculated that statin treatments in smoking patients may provide some beneficial effects on PUFA metabolism in addition to lowering cholesterol levels. PMID- 19128952 TI - Upgrading mixed polyolefin waste with magnetic density separation. AB - Polyolefin fractions are often end fractions resulting from the recycling of end of-life consumer products. Polypropylene (PP) and polyethylene (PE) are present in such fractions as a mixture. For instance, the ratio of PP and PE in car scrap is 70:30 on average. However, the grade of the PP and PE should typically be better than 97% to be reused again as a high quality product. Density separation of the different polyolefins can be a solution. A promising separation technique is the inverse magnetic density separator (IMDS). This paper discusses the potential of shredder residue, one of the possible polyolefin's waste stream sources for the IMDS, in detail. Experiments with the separation of polyolefins with an IMDS prototype show both high grade and high recovery. The paper concludes with the economic opportunities of the IMDS in the recycling of polyolefins. PMID- 19128951 TI - Plasma polyunsaturated fatty acids and regional cerebral glucose metabolism in major depression. AB - Deficiencies in polyunsaturated essential fatty acids (PUFA) are implicated in mood disorders, although mechanisms of action and regional specificity in the brain are unknown. We hypothesized that plasma phospholipid PUFA levels are correlated with regionally specific relative cerebral metabolic rates of glucose (rCMRglu). Medication-free depressed subjects (N=29) were studied using [(18)F] fluoro-2-deoxyglucose positron emission tomography. Docosahexaenoic acid (22:6n 3), arachidonic acid (20:4n-6), and eicosapentaenoic acid (20:5n-3) were assessed as a percentage of total phospholipid PUFA (DHA%, AA%, and EPA%, respectively). DHA% and AA% correlated positively with rCMRglu in temporoparietal cortex. In addition, DHA% correlated negatively with rCMRglu in prefrontal cortex and anterior cingulate. No correlations were seen with EPA%. Thus, under conditions of low plasma DHA, rCMRglu was higher in temporoparietal cortex and lower in anterior cingulate/prefrontal cortex. Opposing effects of DHA on these regions is a hypothesis that could be addressed in future prospective studies with n-3 supplementation. This pilot study is the first to demonstrate fatty acid and regionally specific correlations in the brain between plasma PUFA and rCMRglu in humans. PMID- 19128953 TI - Improving the efficiency of a chemotherapy day unit: applying a business approach to oncology. AB - AIM: To improve the efficiency of a hospital-based chemotherapy day unit (CDU). METHODS: The CDU was benchmarked with two other CDUs to identify their attainable performance levels for efficiency, and causes for differences. Furthermore, an in depth analysis using a business approach, called lean thinking, was performed. An integrated set of interventions was implemented, among them a new planning system. The results were evaluated using pre- and post-measurements. RESULTS: We observed 24% growth of treatments and bed utilisation, a 12% increase of staff member productivity and an 81% reduction of overtime. CONCLUSIONS: The used method improved process design and led to increased efficiency and a more timely delivery of care. Thus, the business approaches, which were adapted for healthcare, were successfully applied. The method may serve as an example for other oncology settings with problems concerning waiting times, patient flow or lack of beds. PMID- 19128954 TI - Evaluation of data quality in the cancer registry: principles and methods Part II. Completeness. AB - The completeness of cancer registry data -- the extent to which all of the incident cancers occurring in the population are included in the registry database -- is an extremely important attribute of a cancer registry. Only a high degree of completeness in case-finding procedures will ensure cancer incidence rates and survival proportions are close to their true value. This second instalment of a two-part review of data quality methods at the cancer registry, focuses on the principles and techniques available for estimating completeness, separating methods into those that are semi-quantitative -- in that they give an indication of the degree of completeness relative to other registries or over time, and more quantitative techniques -- those that provide a numerical evaluation of the extent to which all eligible cases have been registered. PMID- 19128956 TI - The abdominoperineal resection itself is associated with an adverse outcome: the European experience based on a pooled analysis of five European randomised clinical trials on rectal cancer. AB - PURPOSE: The aim of this study is to identify factors associated with the decision to perform an abdominoperineal resection (APR) and to assess if these factors or the surgical procedure itself is associated with circumferential resection margin (CRM) involvement, local recurrence (LR), overall survival (OS) and cancer-specific survival (CSS). PATIENTS AND METHODS: The Swedish Rectal Cancer Trial (SRCT), TME trial, CAO/ARO/AIO-94 trial, EORTC 22921 trial and Polish Rectal Cancer Trial (PRCT) were pooled. A propensity score was calculated, which indicated the predicted probability of undergoing an APR given gender, age and distance, and used in the multivariate analyses. RESULTS: An APR procedure was associated with an increased risk of CRM involvement [odd ratio (OR) 2.52, p<0.001], increased LR rate [hazard ratio (HR) 1.53, p=0.001] and decreased CSS rate (HR 1.31, p=0.002), whereas the propensity score was not. CONCLUSION: The results suggest that the APR procedure itself is a significant predictor for non radical resections and increased risk of LR and death due to cancer for patients with advanced rectal cancer. PMID- 19128957 TI - Axillary metastases in breast cancer patients with negative sentinel nodes: a follow-up of 3548 cases. AB - PREMISES: Sentinel node biopsy (SNB) in patients with breast carcinoma accurately predicts the axillary node status. However, in some 4-7% of patients with negative sentinel nodes, the remaining axillary nodes harbour cancer cells. OBJECTIVE: Our purpose was the long-term observation of a large number of patients who did not receive axillary dissection after a negative sentinel node biopsy, in order to evaluate the incidence of overt axillary metastases. METHODS: Patients (3548) treated from 1996 to 2004, with negative sentinel nodes not submitted to axillary dissection, were followed up to 11 years with a median follow-up of 48 months. RESULTS: Three hundred and sixteen unfavourable events occurred among the 3548 patients, 196 of which (5.5%) related to primary breast carcinoma. Thirty one cases of overt axillary metastases were found (0.9%): they received total axillary dissection and 27 of them are at present alive and well. The 5-year overall survival rate of the whole series was 98%. CONCLUSIONS: Patients with negative sentinel node biopsy not submitted to axillary dissection show, at follow-up, a rate of overt axillary metastases lower than expected. PMID- 19128958 TI - Fermentative production of succinic acid from straw hydrolysate by Actinobacillus succinogenes. AB - In this work, straw hydrolysates were used to produce succinic acid by Actinobacillus succinogenes CGMCC1593 for the first time. Results indicated that both glucose and xylose in the straw hydrolysates were utilized in succinic acid production, and the hydrolysates of corn straw was better than that of rice or wheat straw in anaerobic fermentation of succinic acid. However, cell growth and succinic acid production were inhibited when the initial concentration of sugar, which was from corn straw hydrolysate (CSH), was higher than 60 g l(-1). In batch fermentation, 45.5 g l(-1) succinic acid concentration and 80.7% yield were attained after 48 h incubation with 58 g l(-1) of initial sugar from corn straw hydrolysate in a 5-l stirred bioreactor. While in fed-batch fermentation, concentration of succinic acid achieved 53.2 g l(-1) at a rate of 1.21 g l(-1) h( 1) after 44 h of fermentation. Our work suggested that corn straw could be utilized for the economical production of succinic acid by A. succinogenes. PMID- 19128959 TI - Optimization of adsorption conditions of papain on dye affinity membrane using response surface methodology. AB - The adsorption of papain on Reactive Blue 4 dye-ligand affinity membrane was investigated in a batch system. The combined effects of operating parameters such as initial pH, temperature, and initial papain concentration on the adsorption were analyzed using response surface methodology. The optimum adsorption conditions were determined as initial pH 7.05, temperature 39 degrees C, and initial papain concentration 11.0mg/ml. At optimum conditions, the adsorption capacity of dye-ligand affinity membrane for papain was found to be 27.85 mg/g after 120 min adsorption. The papain was purified 34.6-fold in a single step determined by fast protein liquid chromatography. More than 85% of the adsorbed papain was desorbed using 1.0M NaCl at pH 9.0 as the elution agent. The purification process showed that the dye-ligand immobilized composite membrane gave good separation of papain from aqueous solution. PMID- 19128960 TI - Bioethanol production performance of five recombinant strains of laboratory and industrial xylose-fermenting Saccharomyces cerevisiae. AB - In this study, five recombinant Saccharomyces cerevisiae strains were compared for their xylose-fermenting ability. The most efficient xylose-to-ethanol fermentation was found by using the industrial strain MA-R4, in which the genes for xylose reductase and xylitol dehydrogenase from Pichia stipitis along with an endogenous xylulokinase gene were expressed by chromosomal integration of the flocculent yeast strain IR-2. The MA-R4 strain rapidly converted xylose to ethanol with a low xylitol yield. Furthermore, the MA-R4 strain had the highest ethanol production when fermenting not only a mixture of glucose and xylose, but also mixed sugars in the detoxified hydrolysate of wood chips. These results collectively suggest that MA-R4 may be a suitable recombinant strain for further study into large-scale ethanol production from mixed sugars present in lignocellulosic hydrolysates. PMID- 19128955 TI - The EUROCARE-4 database on cancer survival in Europe: data standardisation, quality control and methods of statistical analysis. AB - This paper describes the collection, standardisation and checking of cancer survival data included in the EUROCARE-4 database. Methods for estimating relative survival are also described. Incidence and vital status data on newly diagnosed European cancer cases were received from 93 cancer registries in 23 countries, covering 151,400,000 people (35% of the participating country population). The third revision of the International Classification of Diseases for Oncology was used to specify tumour topography and morphology. Records were extensively checked for consistency and compatibility using multiple routines; flagged records were sent back for correction. An algorithm assigned standardised sequence numbers to multiple cancers. Only first malignant cancers were used to estimate relative survival from registry, year, sex and age-specific life tables. Age-adjusted and Europe-wide survival were also estimated. The database contains 13,814,573 cases diagnosed in 1978-2002; 92% malignant. A negligible proportion of records was excluded for major errors. Of 5,753,934 malignant adult cases diagnosed in 1995-2002, 5.3% were second or later cancers, 2.7% were known from death certificates only and 0.4% were discovered at autopsy. The remaining 5,278,670 cases entered the survival analyses, 90% of these had microscopic confirmation and 1.3% were censored alive after less than five years' follow-up. These indicators suggest satisfactory data quality that has improved since EUROCARE-3. PMID- 19128961 TI - Effect of pre-aeration and inoculum on the start-up of batch thermophilic anaerobic digestion of municipal solid waste. AB - In this study, a short pre-aeration step was investigated as pre-treatment for thermophilic anaerobic digestion of the organic fraction of municipal solid waste (OFMSW). It was found that pre-aeration of 48 h generated enough biological heat to increase the temperature of bulk OFMSW to 60 degrees C. This was sufficient self-heating of the bulk OFMSW for the start-up of thermophilic anaerobic digestion without the need for an external heat source. Pre-aeration also reduced excess easily degradable organic compounds in OFMSW, which were the common cause of acidification during the start-up of the batch system. Careful consideration however must be taken to avoid over aeration as this consumes substrate, which would otherwise be available to methanogens to produce biogas. To accelerate methane production and volatile solids destruction, the anaerobic digestion in this study was operated as a wet process with the anaerobic liquid recycled through the OFMSW. Appropriate anaerobic liquid inoculum was found to be particularly beneficial. It provided high buffer capacity as well as suitable microbial inoculum. As a result, acidification during start-up was kept to a minimum. With volatile fatty acids (VFAs-acetate in particular) and H2 accumulation typical of hydrolysis and fermentation of the easily degradable substrates during start-up, inoculum with high numbers of hydrogenotrophic methanogens was critical to not only maximise CH4 production but also reduce H2 partial pressure in the system to allow VFAs degradation. In a lab-scale bioreactor, the combined pre-aeration and wet thermophilic anaerobic digestion was able to stabilise the OFMSW within a period of only 12 days. The stabilised inert residual material can be used as a soil amendment product. PMID- 19128962 TI - Influences of organic loading disturbances on the performance of anaerobic filter process to treat purified terephthalic acid wastewater. AB - A lab-scale anaerobic filter process was operated for the treatment of purified terephthalic acid (PTA) wastewater, and the influences of organic loading disturbances on the process performance were investigated. After about 15 month operation, the COD removal efficiency was maintained at 79% under the volumetric loading rate of 5.05 kg-COD/m3/d and the hydraulic retention time (HRT) of 50 h. Interestingly, this performance could be further enhanced over 85% by applying a step-increase/decrease of the HRT, which was mainly due to the increased p toluate degradation. In the shock loading tests of four major pollutants (benzoate, acetate, terephthalate and p-toluate), it was found that the overall process performance was adversely affected by all the shock loadings, indicating that the syntrophic microbial consortium involved in the PTA wastewater treatment is highly sensitive to the organic loading disturbances. The complex inhibition effects of the benzoate and acetate on the terephthalate and p-toluate degradations were mainly responsible for this sensitivity. PMID- 19128963 TI - Advanced swine manure treatment and utilization in Brazil. AB - Animal production has changed from subsistence to an industrial model, lowering production costs but giving rise to higher potential environmental impact. When the effluents are not correctly managed, serious pollution events can occur. In Brazil liquid manure is commonly stored in reception pits or covered lagoons (biodigestors), followed by land application as a biofertilizer. In some regions there is an excess of manure due to low soil support capacities, and in these cases new technologies have to be adopted to export or treat the excess effluent. Manure storage time in pits/covered lagoons and new polymers to separate the solid fraction have been studied in Brazil. Treatment technologies, like swine manure treatment systems (SMTS), have been developed from a technical and economical point of view to optimize the processes and give a technological alternative to pork producers increasing production while reducing environmental impact. PMID- 19128964 TI - Biodiesel waste as source of organic carbon for municipal WWTP denitrification. AB - This paper presents the results of experiments to test biodiesel waste (glycerine -g-phase) as an organic carbon source for the removal of nitrate in a WWTP denitrification process. Investigation of g-phase was first centered on g-phase utilization as an external source for denitrification under laboratory conditions and consequently, after positive results from the laboratory investigation, g phase was applied in the denitrification process in the WWTP Vrutky (35,000 PE). This WWTP had insufficient nitrogen removal via denitrification. Denitrification was insufficient due to an influent with a low BOD5/N ratio (1.7:1) entering into the activated sludge tank. Laboratory experiments and calculations showed that, to reach Ntotal concentration under 10 mg l(-1) in effluent, a biodiesel waste dose of 500 kg(COD) d(-1) was necessary. Glycerol phase (g-phase) dosing into the denitrification tank increased denitrification efficiency by 2.0 - 5.0 mg(NO)(3)( N)l(-1) per 100 l of g-phase dose into the denitrification tank. PMID- 19128965 TI - Partial nitrification to nitrite for treating ammonium-rich organic wastewater by immobilized biomass system. AB - This study focused on the characteristics of the partial nitrification and degradation of organics with immobilized biomass beads in the treatment of ammonium-rich organic wastewater. Sodium alginate (SA) was selected as the best entrapment support after comparing partial nitrification rate and adsorption efficiency. The immobilization methods were optimized by an orthogonal experiment. Zeta position and BET surface area were used to explain the adsorption behavior of SA immobilized beads. FT-IR revealed that a SA immobilized biomass bead was not a simply physical mixture of SA and biomass. The porous structure of SA immobilized biomass beads were observed by scanning electron microscopy (SEM), which confirmed the porosity of the beads. According to the experimental data, the effects of pH and temperature on partial nitrification and COD removal were evidently weakened in SA immobilized biomass beads due to the "protective" effect of immobilization, whereas the effects of HRT and DO were enhanced. PMID- 19128966 TI - Carbonic anhydrase inhibitors: the membrane-associated isoform XV is highly inhibited by inorganic anions. AB - The membrane-associated mouse isozyme of carbonic anhydrase XV (mCA XV), has been investigated for its interaction with anion inhibitors. mCA XV is an isoforms possessing a very particular inhibition profile by anions, dissimilar to that of all other mammalian CAs investigated earlier. Many simple inorganic anions (thiocyanate, cyanide, azide, bicarbonate, hydrogen sulfide, bisulfite and sulfate) showed low micromolar inhibition constants against mCA XV (K(I)s of 8.2 10.1 microM), whereas they acted as much weaker (usually millimolar) inhibitors of other isoforms. Halides, nitrate, nitrite, carbonate, sulfamate, sulfamide and phenylboronic/arsonic acid were weaker inhibitors, with inhibition constants in the range of 27.6-288 microM. Our data may be useful for the design of more potent inhibitors of mCA XV (considering various zinc binding groups present in the anions investigated here, e.g., the sulfonate one) and for understanding some physiologic/pharmacologic consequences of mCA XV inhibition by anions such as bicarbonate or sulfate which show quite high affinity for it. PMID- 19128967 TI - Estradiol dimers as a new class of steroid sulfatase reversible inhibitors. AB - A series of estradiol dimers was synthesized or selected from compounds available in our laboratory and tested for inhibition against steroid sulfatase. Dimers linked by their C17 position, compounds 7 and 8, showed inhibitory potency similar (56% and 54% at 1 microM) to that of our best previously reported reversible inhibitor EM-690 (62% at 1 microM). Docking experiment seems to indicate that C17-C17 dimers bind in a similar way to EM-690 whereas C16-O3 and C16-C16 dimers bind in an upside-down position. PMID- 19128968 TI - Vancomycin resistance: modeling backbone variants with D-Ala-D-Ala and D-Ala-D Lac peptides. AB - To seek vancomycin analogs with broader antibacterial activity, effects of backbone modifications for the agylcon 2 on binding with D-Ala-D-Ala- and D-Ala-D Lac-containing peptides were investigated by Monte Carlo/free energy perturbation (MC/FEP) calculations. The experimental trend in binding affinities for 2 with three tripeptides was well reproduced. Possible modifications of the peptide bond between residues 4 and 5 were then considered, specifically for conversion of the OCNH linkage to CH(2)NH(2)(+) (6), FCCH (7), HCCH (8), and HNCO (9). The MC/FEP results did not yield binding improvements for 7, 8, and 9, though the fluorovinyl replacement is relatively benign. The previously reported analog 6 remains as the only variant that exhibits improved affinity for the D-Ala-D-Lac sequence and acceptable affinity for the D-Ala-D-Ala sequence. PMID- 19128969 TI - Aryl sulphonyl amides as potent agonists of the growth hormone secretagogue (ghrelin) receptor. AB - As part of an on-going lead optimisation effort, a cross screening exercise identified an aryl sulphonyl amide hit that was optimised to afford a highly potent series of ghrelin receptor agonists. PMID- 19128971 TI - Discovery of 2-(alpha-methylbenzylamino) pyrazines as potent Type II inhibitors of FMS. AB - A series of 2-(alpha-methylbenzylamino) pyrazines have shown to be potent inhibitors of the FMS tyrosine receptor kinase. Details of SAR studies, modeling and synthesis of compounds within this series are reported. PMID- 19128970 TI - Synthesis and characterization of a peripherally restricted CB1 cannabinoid antagonist, URB447, that reduces feeding and body-weight gain in mice. AB - Cannabinoid CB(1) receptor antagonists reduce body weight in rodents and humans, but their clinical utility as anti-obesity agents is limited by centrally mediated side effects. Here, we describe the first mixed CB(1) antagonist/CB(2) agonist, URB447 ([4-amino-1-(4-chlorobenzyl)-2-methyl-5-phenyl-1H-pyrrol-3 yl](phenyl)methanone), which lowers food intake and body-weight gain in mice without entering the brain or antagonizing central CB(1)-dependent responses. URB447 may provide a useful pharmacological tool for investigating the cannabinoid system, and might serve as a starting point for developing clinically viable CB(1) antagonists devoid of central side effects. PMID- 19128972 TI - Synthesis and biological activities of high affinity taxane-based fluorescent probes. AB - Three fluorescent probes 3a,3b, and 4 have been synthesized through conjugation of fluorescein and difluorescein groups to the 7-OH of C-2 modified paclitaxel and cephalomannine derivatives with very high affinity to microtubules. All these probes exhibited potent tubulin assembly promotion and tumor cell killing activities, thus may be useful as tools for the determination of thermodynamic parameters and exploration of ligand-microtubule interactions. PMID- 19128973 TI - Corrective postural responses evoked by completely unexpected loss of ground support during human walking. AB - Understanding reactive responses to unexpected perturbation is fundamental to research on falls and their prevention. In this study, seven healthy young males walked along a walkway with and without a completely unexpected loss of ground support that was made by removing a wooden board (6.5 cm high) mounted on the walkway. Electromyography (EMG), ground reaction forces, and knee and ankle kinematics were recorded and comprehensively analyzed to investigate the corrective postural response to the perturbation. Three sequential strategies were observed. First, the fastest response was the reflexive muscle activity of the perturbed ankle, which we argue was evoked and enhanced by the absence of the expected somatosensory afferents at the expected heel contact. We also demonstrated that rapid soleus activity partially contributed to absorbing the impact of the actual touchdown. Second, after the touchdown, we argue that the central nervous system may reset the gait rhythm to permit continued walking by delaying the subsequent take-off. As a result, the duration of the total stance phase was identical to that recorded during normal walking. Third, we observed an adaptive locomotion to surmount the hole; both knees were more flexed than normal in order to allow the subject to withdraw the perturbed leg from the hole. PMID- 19128974 TI - Discovery of N-[(1R,2S,5S)-2-{[(5-chloroindol-2-yl)carbonyl]amino}-5 (dimethylcarbamoyl)cyclohexyl]-5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine 2-carboxamide hydrochloride: a novel, potent and orally active direct inhibitor of factor Xa. AB - In the early 1990's, we reported on the low-molecular selective fXa inhibitor DX 9065a having two amidino groups. However, it had poor oral bioavailability due to its strong basic amidino groups. To obtain fXa inhibitors with improved oral bioavailability, we investigated various non-amidino fXa inhibitors and finally discovered cis-1,2-diaminocyclohexane derivative 4c to have potent fXa inhibition, promising anticoagulant activity, and good oral bioavailability, compared with amidino compound DX-9065a. In addition, we will discuss the influence of the third substituent on the cyclohexane ring on anti-fXa activity, anticoagulant activity, PK profile, and lipophilicity. PMID- 19128975 TI - In vitro solubility, stability and permeability of novel quercetin-amino acid conjugates. AB - In order to discover a quercetin prodrug with improved bioavailability, we synthesized nine quercetin-amion acid conjugates and estimated their pharmacokinetic properties including water solubility, stability against chemical or enzymatic hydrolysis, and cell permeability. Among the synthesized quercetin prodrugs, quercetin-glutamic acid conjugate Qu-E (4g/5g) showed remarkable increases in water solubility, stability, and cell permeability compared with quercetin, which warrants further development as a quercetin prodrug. PMID- 19128976 TI - Synthesis and in vitro anti-proliferative activity of beta-elemene monosubstituted derivatives in HeLa cells mediated through arrest of cell cycle at the G1 phase. AB - Beta-elemene monosubstituted amine, ether and rhenium coordinated complex were synthesized. Their structures were characterized by IR, (1)H NMR, (13)C NMR, HRMS or EA. Their IC(50) on HeLa cell lines, cell cycle and protein expression of G(1) phase (Cyclin D(1), Rb, P-Rb) were detected respectively by the method of WST-1, Flow Cytometry and Western Blot. The Results showed that the in vitro anti proliferative activity of beta-elemene monosubstituted amine and Re(CO)(3)-beta elemene derivatives in human cervix epitheloid carcinoma HeLa cells were improved significantly compared with both of ether derivatives and parent beta-elemene. These derivatives could reduce Rb phosphorylation and cyclin D(1) protein expression to arrest the cell cycle at G(1) phase. PMID- 19128977 TI - Synthesis and biological evaluation of curcuminoid pyrazoles as new therapeutic agents in inflammatory bowel disease: effect on matrix metalloproteinases. AB - Seven N-unsubstituted curcuminoid pyrazoles have been synthesized from the corresponding beta-diketones (including curcumin). We evaluated the possibility of curcuminoid pyrazoles regulating the activity of matrix metalloproteinases (MMPs) by human intestinal epithelial cells in vitro. Zymographic analysis revealed that three compounds significantly down-regulated MMP-9 activity on inflammation-induced intestinal epithelial cells, making them original candidates for the treatment of inflammatory bowel disease (IBD). PMID- 19128978 TI - Synthesis of a bis(2,3-dimethylcyclohexyl-dithiocarbamato)-nitrido 99mTc complex: a potential tracer for myocardial imaging. AB - The (99m)TcN(DMCHDTC)(2) complex, where DMCHDTC is 2,3-dimethylcyclohexyl dithiocarbamato, has been synthesized through a ligand-exchange reaction. The two step procedure involved the initial reaction of (99m)TcO(4)(-) with succinic dihydrazide in the presence of stannous chloride as reducing agent and propylenediamine tetraacetic acid as complexant, followed by the addition of 2,3 dimethylcyclohexyl dithiocarbamate. The radiochemical purity of the complex was over 90%, as measured by thin layer chromatography, without any notable decomposition at room temperature over a period of 6h. The partition coefficient and electrophoresis results indicated that this complex was lipophilic and neutral. Biodistribution in mice showed that the complex accumulated in the heart with high uptake and good retention, the heart uptakes being 12.82, 11.37 and 10.64%ID/g at 5, 30 and 60 min post-injection, respectively. The heart/lung, heart/liver and heart/blood ratios of the complex were 1.06, 0.25 and 8.06 at 60 min post-injection, suggesting it has potential for use as a myocardial imaging agent. PMID- 19128979 TI - Selective separation of iron from uranium in quantitative determination of traces of uranium by alpha spectrometry in soil/sediment sample. AB - During this work, controlled redox potential methodology was adopted for the complete separation of traces of uranium from the host matrix of mixed hydroxide of Iron. Precipitates of Fe(+2) and Fe(+3) along with other transuranic elements were obtained from acid leached solution of soil by raising the pH to 9 with 14N ammonia solution. The concentration of the uranium observed in the soil samples was 200-600 ppb, whereas in sediment samples, the concentration range was 61-400 ppb. PMID- 19128980 TI - Evaluation of scatter-to-primary ratio in radiological conditions. AB - In this work, a Monte Carlo code was constructed taking into account all the relevant photon interactions, including the multiple scattering and interference due to the liquid structure to calculate the secondary to primary ratio for large water samples in radiological conditions. The simulation results were compared to former results and some discrepancies were found mainly for low energies where the elastic scattering plays a major role. PMID- 19128981 TI - Palythine-threonine, a major novel mycosporine-like amino acid (MAA) isolated from the hermatypic coral Pocillopora capitata. AB - Using a high-resolution reverse-phase liquid chromatography method we found that the tissues of the hermatypic coral Pocillopora capitata (collected in Santiago Bay, Mexico) contain a high diversity of primary and secondary mycosporine-like amino acids (MAAs) typical of some reef-building coral species: mycosporine glycine, shinorine, porphyra-334, mycosporine-methylamine-serine, mycosporine methylamine-threonine, palythine-serine, palythine and one additional novel predominant MAA, with an absorbance maximum of 320 nm. Here we document the isolation and characterization of this novel MAA from the coral P. capitata. Using low multi-stage mass analyses of deuterated and non deuterated compounds, high-resolution mass analyses (Time of Flight, TOF) and other techniques, this novel compound was characterized as palythine-threonine. Palythine-threonine was also present in high concentrations in the corals Pocillopora eydouxi and Stylophora pistillata indicating a wider distribution of this MAA among reef building corals. From structural considerations we suggest that palythine threonine is formed by decarboxylation of porphyra-334 followed by demethylation of mycosporine-methylamine-threonine. PMID- 19128982 TI - Differences in binding and effector functions between classes of TNF antagonists. AB - There are currently two Food and Drug Administration-approved classes of biologic agents that target tumor necrosis factor-alpha (TNF-alpha): anti-TNF monoclonal antibodies (mAbs) (adalimumab and infliximab), and soluble TNF receptors (etanercept). This study examined the ability of the TNF antagonists to: (1) bind various polymorphic variants of cell surface-expressed Fc receptors (FcgammaRs) and the complement component C1q, and (2) mediate Ab-dependent cellular cytotoxicity (ADCC) and complement-mediated cytotoxicity (CDC) killing of cells expressing membrane-bound TNF (mTNF) in vitro. Both mAbs and the soluble TNF receptor demonstrated low-level binding to the activating receptors FcgammaRI, FcgammaRIIa, and FcgammaRIIIa, and the inhibitory receptor FcgammaRIIb, in the absence of exogenous TNF. However, upon addition of TNF, the mAbs, but not etanercept, showed significantly increased binding, in particular to the FcgammaRII and FcgammaRIII receptors. Infliximab and adalimumab induced ADCC much more potently than etanercept. In the presence of TNF, both mAbs bound C1q in in vitro assays, but etanercept did not bind C1q under any conditions. Infliximab and adalimumab also induced CDC in cells expressing mTNF more potently than etanercept. Differences in the ability to bind ligand and mediate cell death may account for the differences in efficacy and safety of TNF antagonists. PMID- 19128983 TI - Adiponectin upregulates monocytic activin A but systemic levels are not altered in obesity or type 2 diabetes. AB - Adiponectin is an adipocyte-derived protein with atheroprotective and immunoregulatory function. Adiponectin and activin A reduce foam cell formation and adiponectin activates the p38 MAPK pathway that is well described to induce activin A. Therefore, it was analyzed whether adiponectin alters activin A in primary human monocytes. Adiponectin dose- and time-dependently induced activin A in the supernatant, and the maximal amount was observed after 12h of incubation. Adiponectin-stimulated release of activin A was blocked by a p38 MAPK inhibitor. Metformin and pioglitazone are drugs frequently used to treat diabetic patients and metformin slightly reduced monocytic activin A release whereas pioglitazone had no effect. Type 2 diabetes is associated with elevated inflammatory systemic cytokines but activin A serum levels were similar in slim probands, overweight controls and type 2 diabetic patients. Furthermore, activin A did not correlate to systemic adiponectin, body mass index, waist to hip ratio or C-reactive protein. These findings indicate that adiponectin upregulates monocytic activin A release via the p38 MAPK pathway, and this may in part explain the immunoregulatory and antiatherosclerotic effects of this adipokine. PMID- 19128984 TI - Acute renal dysfunction after cardiac surgery with cardiopulmonary bypass is associated with plasmatic IL6 increase. AB - BACKGROUND: Acute renal dysfunction (ARD) is common after cardiac surgery with cardiopulmonary bypass (CPB). CPB results in a sudden systemic inflammatory response. Systemic and local pro-inflammatory cytokines synthesis has been linked with sub-clinical renal injury, especially tubular lesions. Therefore, we sought to assess the systemic synthesis pro-inflammatory cytokines and its association with perioperative ARD after cardiac surgery with CPB. METHODS: Sixty-two patients undergoing cardiac surgery with CPB were prospectively included. Four groups of patients were defined according to blood creatinine increase: no ARD (less than 25% increase), faint ARD (25-50% increase), moderate ARD (50-100% increase), severe ARD (more than 100% increase). RESULTS: Within the 48 post operative hours was ARD observed as no dysfunction (41.9%), faint (32.2%), moderate (16.1%), severe (9.6%). One patient had to undergo a dialysis. Pre operative characteristics were homogenous between the four groups excepted the left ventricle ejection fraction. ARD was associated with a low urinary output with high sodium excretion fraction. Significant increase of IL-6 level occurred when patients underwent a severe ARD despite no significant differences for the CRP and TNF-alpha concentrations. CONCLUSION: Severe acute renal dysfunction after cardiac surgery with CPB is associated with a significant increased IL-6 systemic production. PMID- 19128985 TI - WITHDRAWN: Artesunate inhibits angiogenesis and induces apoptosis in human umbilical vein endothelial cell. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19128986 TI - Radiographic analysis of commonly prescribed scapular exercises. AB - HYPOTHESIS: The performance of scapular exercises in the context of a clinical model of fatigue will result in superior migration of the humeral head and alterations in shoulder kinematics on radiographic analysis. MATERIALS AND METHODS: Six healthy male volunteers underwent a series of radiographs before and immediately following two scapular exercises. To reproduce a practice setting a clinical model of muscle fatigue was utilized. Radiographs were taken at various degrees of elevation. Glenohumeral angle (GA), scapulothoracic angle (SA), and the position of the humeral head on the glenoid (excursion) were measured on each radiograph. Two-tailed paired sample t-test were used to compare pre-fatigue to post fatigue measures. RESULTS: Following fatigue there was a significant increase in excursion at 45, 90, and 135 degrees of elevation indicating superior migration of the humeral head. There was an increase in SA from 0 to 45 degrees and a decrease in GA from 45 to 90 degrees of elevation. From 90 to 135 degrees an increase in GA was observed. DISCUSSION: The amount of superior migration of the humeral head observed following exercise was consistent with previously reported amounts of superior migration in patients with subacromial impingement. The alterations that occurred in shoulder kinematics following exercise may have resulted from parascapular muscle fatigue as well as rotator cuff muscle insufficiency. CONCLUSION: In the context of a clinical model of fatigue, these scapular exercises resulted in superior migration of the humeral head and alterations in shoulder kinematics on radiographic analysis in healthy subjects. LEVEL OF EVIDENCE: Basic science study. PMID- 19128987 TI - Tuberous sclerosis complex with unilateral perisylvian polymicrogyria and contralateral hippocampal sclerosis - a case report. AB - Malformations of cortical development (MCD) encompass a wide spectrum of brain disorders. Although rare, the combination of certain MCD may occur. We report a rare case of combination of three distinct cerebral pathologies: tuberous sclerosis, polymicrogyria and hippocampal sclerosis in a patient with intractable epilepsy. Ictal EEG and ictal SPECT suggested right perisylvian area as a seizure onset zone. However, on MRI multiple potentially epileptogenic lesions were identified. The neurobiological background of the coexistence of different types of malformations of cortical development remains enigmatic. PMID- 19128988 TI - Repair of full-thickness femoral condyle cartilage defects using allogeneic synovial cell-engineered tissue constructs. AB - OBJECTIVE: Synovium-derived stem cells (SDSCs) have proven to be superior in cartilage regeneration compared with other sources of mesenchymal stem cells. We hypothesized that conventionally passaged SDSCs can be engineered in vitro into cartilage tissue constructs and the engineered premature tissue can be implanted to repair allogeneic full-thickness femoral condyle cartilage defects without immune rejection. METHODS: Synovial tissue was harvested from rabbit knee joints. Passage 3 SDSCs were mixed with fibrin glue and seeded into non-woven polyglycolic acid (PGA) mesh. After 1-month incubation with growth factor cocktails, the premature tissue was implanted into rabbit knees to repair osteochondral defects with Collagraft as a bone substitute in the Construct group. Fibrin glue-saturated PGA/Collagraft composites were used as a Scaffold group. The defect was left untreated as an Empty group. RESULTS: SDSCs were engineered in rotating bioreactor systems into premature cartilage, which displayed the expression of sulfated glycosaminoglycan (GAG), collagen II, collagen I, and macrophages. Six months after implantation with premature tissue, cartilage defects were full of smooth hyaline-like cartilage with no detectable collagen I and macrophages but a high expression of collagen II and GAG, which were also integrated with the surrounding native cartilage. The Scaffold and Empty groups were resurfaced with fibrous-like and fibrocartilage tissue, respectively. CONCLUSION: Allogeneic SDSC-based premature tissue constructs are a promising stem cell-based approach for cartilage defects. Although in vitro data suggest that contaminated macrophages affected the quality of SDSC-based premature cartilage, effects of macrophages on in vivo tissue regeneration and integration necessitate further investigation. PMID- 19128989 TI - Prevalence of digital dermatitis in first lactation cows assessed at breeding cattle auctions. AB - The prevalence of digital dermatitis (DD) in first lactation cows (FLCs) presented at 10 breeding cattle auctions was determined in a single auction centre in Austria over a period of 14 months. At each auction, FLCs were selected randomly for claw examination from the monthly auction catalogue. After obtaining owner consent, the hindclaws were examined in a walk-in crush. The type of lesion (M1-M4) and the diameter of DD lesions together with claw horn lesions and the trimming status of the claws were evaluated. From a total of 1110 FLCs registered for the ten auctions, 399 (36%) were randomly chosen for examination and 199 FLCs (17.9%) could be defined. In 63 cows (15.7%), the owners refused consent. A total of 24 FLCs were found to have DD lesions on one or both hindlimbs, resulting in a DD prevalence of 12.1%. The size of lesions ranged from 0.5-3 cm in diameter. No influence of environmental factors (such as season or housing) on the prevalence of DD could be found. In 30/199 (15.1%) of FLCs examined, claw horn lesions such as double sole, white line disease and sole ulcer were observed, but both claw horn and DD lesions were seen in only three cows. Potential buyers at auctions should be aware of both conditions, as the 12.1% prevalence of DD and the 15.1% prevalence of claw horn lesions in FLCs are considerable. Critical evaluation of claw health and DD is essential when purchasing breeding cattle. PMID- 19128990 TI - The dipeptidyl peptidase IV inhibitor NVP-DPP728 reduces plasma glucagon concentration in cats. AB - Glucagon-like peptide-1 (GLP-1) analogues and inhibitors of its degrading enzyme, dipeptidyl peptidase IV (DPPIV), are interesting therapy options in human diabetics because they increase insulin secretion and reduce postprandial glucagon secretion. Given the similar pathophysiology of human type 2 and feline diabetes mellitus, this study investigated whether the DPPIV inhibitor NVP-DPP728 reduces plasma glucagon levels in cats. Intravenous glucose tolerance tests (ivGTT; 0.5 g/kg glucose after 12 h fasting) and a meal response test (test meal of 50% of average daily food intake, offered after 24 h fasting) were performed in healthy experimental cats. NVP-DPP728 (0.5-2.5 mg/kg i.v. or s.c.) significantly reduced glucagon output in all tests and increased insulin output in the ivGTT. Follow-up studies will investigate the potential usefulness as therapy in diabetic cats. PMID- 19128991 TI - Alexander disease: early presence of cerebral MRI criteria. AB - Alexander disease is a rare neurodegenerative disorder. Its most frequent subtype, the infantile form, is characterized by an early onset and a rapid neurological deterioration during the first months of life. Since the publication of cerebral radiological criteria in 2001, the disease has often been recognized by magnetic resonance imaging (MRI) findings. We report the case of a girl who at the age of 3 months presented with partial seizures and a normal neurological examination. MRI revealed the presence of a periventricular rim, extensive frontal white matter abnormalities, abnormalities of the basal ganglia and thalami and contrast enhancement involving optic chiasm, fornix, hypothalamus and mamillary bodies, corresponding to four of the five reported MRI criteria for Alexander disease. Additional MRI abnormalities not described so far were also observed. The diagnosis was confirmed by genetic analysis. This case illustrates that diagnostic MRI abnormalities of Alexander disease may be present at a very young age, long before the appearance of characteristic clinical signs. Early diagnosis by MRI allows prompt counselling of families. PMID- 19128992 TI - Resolution enhancement in MRI of laser polarized 3He by control of diffusion. AB - Diffusion of atoms or molecules in presence of magnetic field gradients not only attenuates the NMR signal but also leads to distortions close to restricting boundaries. This phenomenon is most evident in imaging with laser polarized (LP) noble gases. Diffusion of gases can be manipulated, however, by admixing inert gases of different molecular weight. In this work we analyze the effect of mixing LP-(3)He with SF(6) on the image quality of a phantom consisting of an arrangement of capillaries with different diameters. Admixing buffer gases of higher molecular weight changes the contrast and offers a means to record images with high spatial and time resolution. Additionally we demonstrate how distortions due to edge enhancement can be reduced even for long timed MRI sequences. PMID- 19128993 TI - Nano-emulsions of fluorinated trityl radicals as sensors for EPR oximetry. AB - This article reports the development and evaluation of two nano-emulsions (F45T 03/HFB and F15T-03/PFOB) containing fluorinated trityl radicals dissolved in perfluorocarbons. Preparation with a high-pressure homogenizer conferred sub micronic size to both nano-emulsions. In vitro and in vivo EPR spectroscopy showed that the nano-emulsions had much greater oxygen sensitivity than the hydrophilic trityl, CT-03. In vivo experiments in rodents confirmed the ability of the nano-emulsions to follow the changes in oxygen concentration after induced ischemia. Histological evaluation of the tissue injected with the nano-emulsions revealed some acute toxicity for the F45T-03/HFB nano-emulsion but none for the F15T-03/PFOB nano-emulsion. These new formulations should be considered for further EPR oximetry experiments in pathophysiological situations where subtle changes in tissue oxygenation are expected. PMID- 19128994 TI - Isomerization and electronic relaxation of azobenzene after being excited to higher electronic states. AB - In this work, some critical structures (e.g. stable structure, transition state, local minimum and conical intersection) of azobenzene photoisomerization were optimized by means of ab initio CASSCF calculation. The potential energy surfaces for the CNNC dihedral torsion and CNN bond angle concerted-inversion pathway were mapped to explore the relaxation process of azobenzene (AB) photoisomerization. The results indicate that the rotational mechanism favors the photoisomerization of the S(1)(n,pi*) and S(2)(pi,pi*) trans-AB. The concerted-inversion mechanism may operate in the decay process of S(2)(pi,pi*) or higher state trans-AB. By borrowing the (n,pi*; pi,pi*) and (n(2),pi*(2)) electronic states, trans-AB upon excitation to the higher states can quickly relax to the S(1)(n,pi*) or ground state via the rotation or concerted-inversion pathway. The forming ground-state species with higher vibrational energy from the higher excited states will become the stable trans-isomer through the concerted-inversion pathway. These relaxation processes have been confirmed by the conical intersections calculated by the high level CASSCF method. PMID- 19128995 TI - Dosimetric characteristics of a MOSFET dosimeter for clinical electron beams. AB - The fundamental dosimetric characteristics of commercially available metal oxide semiconductor field effect transistor (MOSFET) detectors were studied for clinical electron beam irradiations. MOSFET showed excellent linearity against doses measured using an ion chamber in the dose range of 20-630cGy. MOSFET reproducibility is better at high doses compared to low doses. The output factors measured with the MOSFET were within +/-3% when compared with those measured with a parallel plate chamber. From 4 to 12MeV, MOSFETs showed a large angular dependence in the tilt directions and less in the axial directions. MOSFETs do not show any dose-rate dependence between 100 and 600MU/min. However, MOSFETs have shown under-response when the dose per pulse of the beam is decreased. No measurable effect in MOSFET response was observed in the temperature range of 23 40 degrees C. The energy dependence of a MOSFET dosimeter was within +/-3.0% for 6-18MeV electron beams and 5.5% for 4MeV ones. This study shows that MOSFET detectors are suitable for dosimetry of electron beams in the energy range of 4 18MeV. PMID- 19128996 TI - Salivary gland transcriptome analysis during Plasmodium infection in malaria vector Anopheles stephensi. AB - BACKGROUND: Understanding the tissue-specific molecular cross-talk mechanism during the mosquito-parasite interaction is of prime importance in the design of new strategies for malaria control. Because mosquito salivary glands are the final destination for the parasite maturation and transmission of vector-borne diseases, identification and characterization of salivary genes and their products are equally important in order to access their effect on the infectivity of the parasite. During the last five years there have been several studies on the sialomes of Anopheles mosquitoes, however very limited information is available on the changes in the salivary gland transcriptome in the presence of Plasmodium, and this information is limited to the mosquito Anopheles gambiae. METHODS: In this study we aimed to explore and identify parasite-induced transcripts from the salivary glands of Anopheles stephensi, using a subtractive hybridization protocol. RESULTS: Ninety-four percent of expressed sequence tags (ESTs) showed close homology to previously known families of mosquito salivary gland secretary proteins, representing the induced expression of alternative splicing and/or additional new members of the protein family. The remaining 6% of ESTs did not yield significant homology to any known proteins in the non redundant database and thus may represent a class of unknown/novel salivary proteins. Primary analysis of the ESTs also revealed identification of several novel immune-related transcripts, including defensin and cecropins, probably involved in counter-activation of the antagonistic defense system. A comprehensive description of each family of proteins has been discussed in relation to the tissue-specific mosquito-parasite interaction. CONCLUSION: This is the first report on the identification of new putative salivary genes, presumably activated during parasite infection. PMID- 19128997 TI - Use of serum antistreptolysin O titers in the microbial diagnosis of orthopedic infections. AB - The utility of serologic tests in the microbial diagnosis of orthopedic infections is unknown. Antistreptolysin O titer determination is inexpensive and accurate in the diagnosis of beta-hemolytic group A, C, and G streptococci. In patients with negative culture results and positive titers, antibiotics might be reduced to the narrowest spectrum, penicillin. PMID- 19128998 TI - [Endometriosis and pelvic pain: epidemiological evidence of the relationship and implications]. AB - The relationship between chronic pelvic pain symptoms and endometriosis is unclear because painful symptoms are frequent in women without this pathology, and because asymptomatic forms of endometriosis exist. Our comprehensive review attempts to clarify the links between the characteristics of lesions and the semiology of chronic pelvic pain symptoms. Based on randomized trials against placebo, endometriosis appears to be responsible for chronic pelvic pain symptoms in more than half of confirmed cases. A causal association between severe dysmenorrhoea and endometriosis is very probable. This association is independent of the macroscopic type of the lesions or their anatomical locations and may be related to recurrent cyclic microbleeding in the implants. Endometriosis-related adhesions may also cause severe dysmenorrhoea. There are histological and physiopathological arguments for the responsibility of deeply infiltrating endometriosis (DIE) in severe chronic pelvic pain symptoms. DIE-related pain may be in relation with compression or infiltration of nerves in the subperitoneal pelvic space by the implants. The painful symptoms caused by DIE present particular characteristics, being specific to involvement of precise anatomical locations (severe deep dyspareunia, painful defecation) or organs (functional urinary tract signs, bowel signs). They can thus be described as "location indicating pain". A precise semiological analysis of the chronic pelvic pain symptoms characteristics is useful for the diagnosis and therapeutic. PMID- 19128999 TI - Sonolysis of levodopa and paracetamol in aqueous solutions. AB - Pharmaceutical products are often present in wastewater treatment effluents, rivers, lakes and, more rarely, in groundwater. The advanced oxidation methods, like ultrasound, find a promising future in the area of wastewater treatment. The aim of this paper is to evaluate the influence of several parameters of the ultrasound process on the degradation of paracetamol, a widely used non-steroidal anti-inflammatory recalcitrant drug found in water and levodopa, the most frequently prescribed drug for the treatment of Parkinson disease. Experiments were carried out at 574, 860 and 1134kHz of ultrasonic frequency with horn-type sonicator and actual power values of 9, 17, 22 and 32W at 20 degrees C. Initial concentrations of 25, 50, 100 and 150mgL(-1) of both products were used. Treatment efficiency was assessed following changes in pharmaceuticals concentration and chemical oxygen demand. The sonochemical degradation of both products follows a pseudo-first-order reaction kinetics. Complete removal of pharmaceuticals was achieved in some cases but some dissolved organic carbon remains in solution showing that long lived intermediates were recalcitrant to ultrasound irradiation. Pollutants conversion and COD removal were found to decrease with increasing the initial solute concentration and decreasing power. The best results were obtained with 574kHz frequency. Investigations using 1 butanol as radical scavenger and H(2)O(2) as promoter revealed that pollutants degradation proceeds principally through radical reactions, although some differences were observed between both molecules. Addition of H(2)O(2) had a positive effect on degradation rate, but the optimum concentration of hydrogen peroxide depends on the pollutant. PMID- 19129000 TI - The Windkessel model revisited: a qualitative analysis of the circulatory system. AB - In this paper, we derive a comprehensive computational model to estimate the arterial pressure and the cardiac output of humans, by refining and adapting the well-established equations of the Windkessel theory. The model inputs are based on patient specific factors such as age, sex, smoking and fitness habits as well as the use of specific drugs. The model's outputs correlate very strongly with physiological observations, with a low error of approximately 5% for the arterial pressure. PMID- 19129001 TI - The influence of plaque composition on underlying arterial wall stress during stent expansion: the case for lesion-specific stents. AB - Intracoronary stent implantation is a mechanical procedure, the success of which depends to a large degree on the mechanical properties of each vessel component involved and the pressure applied to the balloon. Little is known about the influence of plaque composition on arterial overstretching and the subsequent injury to the vessel wall following stenting. An idealised finite element model was developed to investigate the influence of both plaque types (hypercellular, hypocellular and calcified) and stent inflation pressures (9, 12 and 15 atm) on vessel and plaque stresses during the implantation of a balloon expandable coronary stent into an idealised stenosed artery. The plaque type was found to have a significant influence on the stresses induced within the artery during stenting. Higher stresses were predicted in the artery wall for cellular plaques, while the stiffer calcified plaque appeared to play a protective role by reducing the levels of stress within the arterial tissue for a given inflation pressure. Higher pressures can be applied to calcified plaques with a lower risk of arterial vascular injury which may reduce the stimulus for in-stent restenosis. Results also suggest that the risk of plaque rupture, and any subsequent thrombosis due to platelet deposition at the fissure, is greater for calcified plaques with low fracture stresses. PMID- 19129002 TI - The importance of position and path repeatability on force at the knee during six DOF joint motion. AB - Mechanical devices, such as robotic manipulators have been designed to measure joint and ligament function because of their ability to position a diarthrodial joint in six degrees-of-freedom with fidelity. However, the precision and performance of these testing devices vary. Therefore, the objective of this study was to determine the effect of systematic errors in position and path repeatability of two high-payload robotic manipulators (Manipulators 1 and 2) on the resultant forces at the knee. Using a porcine knee, the position and path repeatability of these manipulators were determined during passive flexion extension with a coordinate measuring machine. The position repeatability of Manipulator 1 was 0.3 mm in position and 0.2 degrees in orientation while Manipulator 2 had a better position repeatability of 0.1 mm in position and 0.1 degrees in orientation throughout the range of positions examined. The corresponding variability in the resultant force at the knee for these assigned positions was 32+/-33 N for Manipulator 1 and 4+/-1 N for Manipulator 2. Furthermore, the repeatability of the trajectory of each manipulator while moving between assigned positions (path repeatability) was 0.8 mm for Manipulator 1 while the path repeatability for Manipulator 2 was improved (0.1 mm). These path discrepancies produced variability in the resultant force at the knee of 44+/-24 and 21+/-8 N, respectively, for Manipulators 1 and 2 primarily due to contact between the articular surfaces of the tibia and femur. Therefore, improved position and path repeatability yields lower variability in the resultant forces at the knee. Although position repeatability has been the most common criteria for evaluating biomechanical testing devices, the current study has clearly demonstrated that path repeatability can have an even larger effect on the variability in resultant force at the knee. Consequently, the repeatability of the path followed by the joint throughout its prescribed trajectory is as important as the repeatability of the joint at reaching positions making up its trajectory, particularly when joint contact occurs. PMID- 19129003 TI - Analysis of natural and artificial ultramarine blue pigments using laser induced breakdown and pulsed Raman spectroscopy, statistical analysis and light microscopy. AB - Pulsed laser induced breakdown spectroscopy (LIBS) and Raman spectroscopy were performed using a novel laboratory setup employing the same Nd:YAG laser emission at 532 nm for the analysis of five commercially available pigments collectively known as "ultramarine blue", a sodium silicate material of either mineral origin or an artificially produced glass. LIBS and Raman spectroscopy have provided information regarding the elemental and molecular composition of the samples; additionally, an analytical protocol for the differentiation between natural (lapis lazuli) and artificial ultramarine blue pigments is proposed. In particular LIBS analysis has allowed the discrimination between pigments on the basis of peaks ascribed to calcium. The presence of calcite in the natural blue pigments has been confirmed following Raman spectroscopy in specific areas of the samples, and micro-Raman and optical microscopy have further corroborated the presence of calcite inclusions in the samples of natural origin. Finally multivariate analysis of Laser induced breakdown spectra using principal component analysis (PCA) further enhanced the differentiation between natural and artificial ultramarine blue pigments. PMID- 19129004 TI - A study of the structure of the pindolol based on infrared spectroscopy and natural bond orbital theory. AB - Beta-adrenoceptor-blocking agents (beta-blockers) are on the list of the top selling drugs. Pindolol is a representative of this type of compound, either from the structural point of view, or as reference for comparison of the pharmacokinetic properties of the beta-blockers. A study of the pindolol structure based on infrared spectroscopy and natural bond orbital (NBO) theory is the main aim of the present research. FTIR spectra of the solid pindolol were recorded from 4000 to 400cm(-1), at temperatures between 25 and -170 degrees C. For spectral interpretation, the theoretical vibrational spectra of the conformer present in the solid was obtained at the B3LYP/6-31G* level of theory. NBO analysis of the reference conformer, before and after optimization, was carried out at the same level of theory referred above. Characteristic absorption vibrational bands of the spectra of solid pindolol and of the isolated conformer were identified. Intra- and intermolecular interactions in pindolol were confirmed by the frequency shift of the vibrational modes and by the NBO theory. A detailed molecular picture of pindolol and of its intermolecular interactions was obtained from spectroscopy and NBO theory. The combination of both methods gives a deeper insight into the structure. PMID- 19129005 TI - Experimental investigation on large-area dielectric barrier discharge in atmospheric nitrogen and air assisted by the ultraviolet lamp. AB - In this paper, ultraviolet radiation produced by the ultraviolet lamp is employed to supply pre-ionization for the dielectric barrier discharge in N(2) or air at atmospheric pressure. The effect of the ultraviolet pre-ionization on improving the uniformity of the dielectric barrier discharge is investigated experimentally. The atmospheric pressure glow discharge of the large area (270 mm x 120 mm) is obtained successfully via the ultraviolet pre-ionization in atmospheric DBD in N(2) when the gas gap decrease to 3mm. Based on the emission spectra, the mechanism which ultraviolet pre-ionization improves the uniformity of the dielectric barrier discharge is discussed. PMID- 19129006 TI - Influence of Zn-Cd substitution: spectroscopic and theoretical investigation of 8 hydroxyquinoline complexes. AB - It is now well known that zinc is crucial for the synthesis of nucleic acids and, consequently, for cellular division. However, (67)Zn, the NMR-detectable isotope, is one of the isotopes most poorly studied by NMR. The strategy used for NMR studies is the substitution of Zn by (113)Cd. In this work, we employed (13)C, (113)Cd NMR (CPMAS), X-ray and DFT calculation in order to evaluate the Zn-Cd substitution using 8-hydroxyquinoline like prototype compound. Our results show that there are strong structural and electronics effects are involved in the substitution. PMID- 19129007 TI - Photosensitization of SnO(2)/ZnO semiconductors with zinc-phthalocyanine. AB - Zinc-phthalocyanine with tyrosine substituent (ZnPcTyro) was attached to different nanocrystalline semiconductors (SnO(2) and ZnO) via carboxylic acid group, the interaction of zinc-phthalocyanine with colloidal ZnO and SnO(2) was studied by absorption and fluorescence spectroscopy. The apparent association constant for the association between ZnPcTyro and SnO(2)/ZnO is ranged from (3.7+/-0.2)x10(5)M(-1) to (6.7+/-0.2)x10(4)M(-1) with a degree of association ranged from 85% to 95%. The presence of the tyrosine group in ZnPcTyro affects solubility of the dye and affects the aggregation behavior of the dye. Anchoring of the dye complex to the nanocrystalline semiconductors enables ultrafast injection of electrons from the excited state into the conduction band of semiconductors. Maximum incident photon-to-current conversion of 0.84-1% at 600 nm and photon-to-current conversion efficiency of around 37% and 43% was obtained for ZnPc-sensitized-SnO(2) and ZnPc-sensitized-ZnO. PMID- 19129008 TI - Papular-purpuric gloves and socks syndrome associated with B19V infection in a 6 year-old child. AB - A 6 year-old girl was admitted for evaluation of a fever associated with a petechial rash of 2 days' duration. She was in good general condition with no acute distress. Inspection of the skin revealed an amazing papular and purpuric rash of predominantly acral and symmetrical distribution and sharply demarcated on the ankles. All laboratory tests were found normal. Rash and fever completely resolved in less than 3 days. Serological testing for parvovirus B19 (B19V) antibodies was positive for IgM but negative for IgG. Moreover, B19V DNA was detected in serum with a viral load of 2.24 x 10(8) copies per mL. So we concluded of a paediatric case of popular-purpuric gloves and socks syndrome (PPGSS) associated with B19V infection. PPGSS is an idiosyncratic reaction to viral infection. The syndrome has been associated with several viruses such as HHV6, measles, coxsackie B6, and above all B19V. PPGSS occurs mostly in young adults. It is characterised by a typical papular and purpuric rash with an acral distribution and a sharp demarcation on the wrists and ankles. The rash is often pruritic and can be accompanied by mucosal lesions and/or systemic symptoms such as fever, asthenia and lymphadenopathy. Most of the time, the disease is self limited with a short course and a benign prognosis. A very similar disease has been described in some children. The distinctive clinical characteristics of PPGSS in children should be recognized by paediatrician in particular at the emergency room in order to avoid superfluous explorations. PMID- 19129009 TI - Merkel cell polyomavirus is not detected in prostate cancers, surrounding stroma, or benign prostate controls. AB - BACKGROUND: Merkel cell polyomavirus (MCV or MCPyV) is a recently discovered human polyomavirus that is implicated in the pathogenesis of Merkel cell carcinoma (MCC). Although the transmission route for MCV is not yet known, other polyomaviruses, such as BKV, cause non-malignant pathology in the urinary tract. Like MCC, prostate cancer predominantly affects the elderly. Furthermore, prostate cancers and premalignant precursors exhibit chronic inflammation, which suggests a possible infectious involvement. We therefore examined whether MCV might participate in the pathogenesis of prostate cancer. OBJECTIVE: To determine the presence of MCV RNA in prostate cancer and surrounding stroma or normal prostate tissue. STUDY DESIGN: RNA was extracted from 28 patient-matched cancerous and 28 benign prostate epithelial samples, and six additional cancer adjacent stromal samples. All tissues were laser-capture micro-dissected. DNA and RNA from a sequence-verified MCV-containing MCC tumor served as a positive control. Quantitative reverse-transcription PCR was used to assess the presence or absence of MCV T antigen transcript. RESULTS: No MCV T antigen was detected in prostate carcinomas, patient-matched benign samples, or tumor-adjacent stroma, with appropriate sensitivity of the assay demonstrated by an MCC tumor. CONCLUSIONS: MCV infection appears unlikely to be a significant factor in prostate carcinogenesis and there is no evidence of the prostate serving as a reservoir for MCV. PMID- 19129010 TI - Biomechanical rigidity of an all-polyetheretherketone anterior thoracolumbar spinal reconstruction construct: an in vitro corpectomy model. AB - BACKGROUND CONTEXT: Polyetheretherketone (PEEK) is gaining favor as a spinal implant material for interbody and corpectomy cages as well as stabilizing rods. However, there has been little correlation to a relevant and reproducible clinical model. Biomechanical data on PEEK rod constructs have not been reported. PURPOSE: To quantify the stabilizing effects of PEEK versus titanium (Ti) instrumentation in a thoracolumbar corpectomy model. STUDY DESIGN: Corpectomy and randomized instrumentation with an all-Ti, all-PEEK, and hybrid cage/rod construct were performed on cadaveric spines to assess biomechanical differences. METHODS: Pure unconstrained bending moments were applied to the intact spine and subsequent test constructs in the three physiologic planes using a load control protocol. Motion tracking and analysis were carried out to quantify and compare the range of motion (ROM) between different test constructs in each plane. RESULTS: Flexion ROM did not show significant changes compared with intact, whereas the all-Ti and hybrid construct reduced ROM significantly in extension. Lateral bending was significantly reduced in all the treatment groups. Rotational stability of the construct was significantly compromised by an all-PEEK spinal construct. CONCLUSION: The rigidity of the corpectomy construct increased as the amount of Ti in the construct increased. A hybrid construct incorporating a PEEK corpectomy cage and Ti rods may provide adequate stability for an anterior thoracolumbar reconstruction in the sagittal and coronal planes. An all-PEEK construct may provide adequate stability in the coronal and sagittal planes but may compromise the stability significantly in axial rotation. Consideration should be given for supplemental posterior instrumentation if an all-PEEK construct is used in an anterior thoracolumbar spinal reconstruction procedure. PMID- 19129011 TI - Determination of vitamin K1 in plasma by solid phase extraction and HPLC with fluorescence detection. AB - We describe a procedure for quantification of vitamin K(1) in human plasma by HPLC. Samples, enriched with a vitamin K derivative as internal standard, were deproteinized, purified on polymeric RP-SPE cartridges and injected into HPLC equipped with a post-column on-line zinc metal reactor and a fluorometric detector. Median level in blood donors (n=87) was 1.967 nmol/L (0.93-4.01, 5th 95th percentiles), with a significant correlation between plasma levels and age (r=0.276, p=0.00958) and a lower (not significant) value in women than in men. This method, easy-to-handle and with a high throughput, can be used to identify covert states of vitamin K intake deficiency in patients thus at risk of alterations in blood clotting or bone mineralization. PMID- 19129012 TI - WITHDRAWN: Rational reformulation of long-acting preparations: the development of a novel formulation of prolonged-release lanreotide. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19129013 TI - Peptide modification of polyethersulfone surfaces to improve adipose-derived stem cell adhesion. AB - Polyethersulfone (PES) is a nondegradable, biocompatible, synthetic polymer that is commonly utilized as a membrane material for applications such as hemodialysis, ultrafiltration and bioreactor technology. Various studies have shown surface modification to be a valuable tool in the development of nondegradable materials which promote cell adhesion. Cells of interest include adipose-derived stem cells (ASCs). ASCs are multipotent mesenchymal stem cells that are useful for various regenerative medicine applications. In this study, we hypothesized that PES surfaces modified with a peptide sequence based from fibronectin, such as Arg-Gly-Asp (RGD), Arg-Gly-Asp-Ser and Gly-Arg-Gly-Asp-Ser, would increase ASC adhesion compared to unmodified PES surfaces. The synthetic peptides were covalently bonded to amine-modified PES surfaces using 1-ethyl-3 (dimethylaminopropyl) carbodiimide. The surfaces were characterized using a ninhydrin assay and contact angle measurements. The ninhydrin assay confirmed the presence of amine groups on the surface of peptide-treated PES disks. Advancing water contact angles were analyzed to detect changes in the hydrophilicity of the polymer surfaces, and results indicated our PES membranes had excellent hydrophilicity. The attachment and proliferation of human ASCs was assessed and RGD-treated surfaces resulted in a higher number of attached ASCs after 6 and 48 h, as compared to unmodified PES surfaces. Additionally, varying concentrations of the RGD peptide sequence concentration were examined. These results indicate that PES membranes modified with the RGD peptide sequence can be utilized for enhanced ASC attachment in biomedical applications. PMID- 19129014 TI - Two-stage treatment for diabetic foot: surgical peripheral revascularization and minor amputation in day-surgery admission. AB - Diabetic foot is complex and difficult to treat. More aggressive treatment using peripheral distal by-pass frequently combined to minor amputations has greatly improved limb salvage in most patients. However, diabetes-related amputations are at high risk of non-healing or superinfection, thus requiring a second-step surgical revision treatment more frequently than in non-diabetic patients. Several advanced technologies have been developed to improve the treatment of diabetic foot wounds including Vacuum Assisted Therapy: we present 3 cases of diabetic patients treated with preliminary surgical peripheral revascularization, subsequent minor amputation in combination with Vacuum Assisted Therapy performed in a day-surgery regime. PMID- 19129015 TI - Secondary antireflux surgery. AB - BACKGROUND & AIM: With the introduction of laparoscopy, the number of antireflux surgeries (ARS), and consequently failed fundoplications, had increased. We report the mechanisms of fundoplication failure after ARS, and present our experience in surgical correction of failure. METHODS: Twenty nine patients who had failed fundoplication were reoperated. Pre- and postoperative evaluation of patients included symptom severity score, endoscopy, barium study, esophageal motility and pH metry. RESULTS: The initial procedures were Nissen in 16, Toupet in 10, and Nissen-Rossetti in 3 patients. The causes of failure were transdiaphragmatic migration of fundoplication (n=7), disrupted fundoplication (n=7), tight fundoplication (n=4), slipped fundoplication (n=3), paraesophageal herniation (n=3), tight crural repair (n=3), and migration with disruption (n=2). The secondary ARS performed were Nissen (n=16), Toupet (n=9), paraesophageal hernia repair with crural repair (n=2), widening of crural repair (n=1), and taking down fundoplication (n=1). Per- (n=4) and postoperative (n=5) complications were minor with no mortality. At Follow-up, symptoms were significantly improved. CONCLUSION: Reoperations for failed ARS may be performed safely with excellent results. Proper patient selection and paying attention to some technical details at initial ARS could safe the patient another surgery. PMID- 19129016 TI - The effect of centrifugation on viability of fat grafts: an evaluation with the glucose transport test. AB - BACKGROUND: An up-to-date, simple, but useful technique to evaluate the viability of fat grafts prior to transplant is lacking. The purpose of this study is to introduce the glucose transport test - a new method to evaluate the viability of fat grafts after they are subjected to different centrifugal forces in vitro. METHOD: Fat grafts were harvested from healthy patients who underwent liposuction for body contouring. The glucose transport test was performed to evaluate the viability of fat grafts after centrifugation with different forces (1000-4000 rpm). An MTT assay was also performed with the same experimental protocol for comparison. Routine histological examination was done in all groups to examine possible structural destruction after centrifugation. RESULTS: When compared with the group not subjected to centrifugation, the glucose transport test showed a significant decrease in viability of fat grafts in all of the other four groups (all p<0.001). There was a linear reduction of viability in fat grafts with the increase in centrifugal force (all p<0.03). MTT assay showed similar findings on the viability of fat grafts in all five groups and correlated well with the glucose transport test (r=0.9870). Histology showed significantly distorted and fractured adipocytes when the centrifugal force reached 4000 rpm. CONCLUSION: Our study demonstrates the harmful effect on the viability of fat grafts with an increase in centrifugal force and, for the first time, that the glucose transport test may be an effective and potentially useful method to evaluate the viability of fat grafts in a clinical setting. PMID- 19129017 TI - Determining the presence of bias error using statistical methods. AB - Current techniques in image-guided surgery rely on the use of localizers for the measurement of position in physical space. These measurements are prone to error due to intrinsic properties of the localizer used. The error and thus accuracy of a localizer can be determined using various techniques, many of which assume that the error is isotropic and free of bias. A bias error adds an orientation dependence to the error of measured points. Determination of the presence of a bias error is an important component in the characterization of a localizer's performance. Statistical analysis of localized points on a rigid phantom can be used to detect the presence of a bias error. In this paper, we will examine the use of statistical techniques in the characterization of a series of localizers and how that information is useful in determining localizer efficacy. PMID- 19129018 TI - Facial recognition from volume-rendered magnetic resonance imaging data. AB - Three-dimensional (3-D) reconstructions of computed tomography (CT) and magnetic resonance (MR) brain imaging studies are a routine component of both clinical practice and clinical and translational research. A side effect of such reconstructions is the creation of a potentially recognizable face. The Health Insurance Portability and Accountability Act of 1996 (HIPAA) Privacy Rule requires that individually identifiable health information may not be used for research unless identifiers that may be associated with the health information including "Full face photographic images and other comparable images ..." are removed (de-identification). Thus, a key question is: Are reconstructed facial images comparable to full-face photographs for the purpose of identification? To address this question, MR images were selected from existing research repositories and subjects were asked to pair an MR reconstruction with one of 40 photographs. The chance probability that an observer could match a photograph with its 3-D MR image was 1 in 40 (0.025), and we considered 4 successes out of 40 (4/40, 0.1) to indicate that a subject could identify persons' faces from their 3-D MR images. Forty percent of the subjects were able to successfully match photographs with MR images with success rates higher than the null hypothesis success rate. The Blyth-Still-Casella 95% confidence interval for the 40% success rate was 29%-52%, and the 40% success rate was significantly higher ( P < 0.001) than our null hypothesis success rate of 1 in 10 (0.10). PMID- 19129019 TI - Investigating the interaction between oncogene and tumor suppressor protein. AB - It is known that cancer develops when cells in a part of the body begin to grow out of control. Because cancer cells continue to grow and divide with no order, they never differentiate into the specific tissue, and thus, they are functionally different from normal cells. However, there are some genes that help to prevent cells' malignant behavior, and therefore, are referred to as tumor suppressor genes. Here, we have investigated the structural and functional relationships of p53, oncogene and interleukin 2 (IL2) proteins using the resonant recognition model (RRM), a physico-mathematical approach based on digital signal processing methods. In addition, using the RRM concepts, we have designed the peptide analoges that would exhibit tumor-suppression-like activity and be used in anticancer vaccine development. PMID- 19129020 TI - Enhancing privacy and authorization control scalability in the grid through ontologies. AB - The use of data Grids for sharing relevant data has proven to be successful in many research disciplines. However, the use of these environments when personal data are involved (such as in health) is reduced due to its lack of trust. There are many approaches that provide encrypted storages and key shares to prevent the access from unauthorized users. However, these approaches are additional layers that should be managed along with the authorization policies. We present in this paper a privacy-enhancing technique that uses encryption and relates to the structure of the data and their organizations, providing a natural way to propagate authorization and also a framework that fits with many use cases. The paper describes the architecture and processes, and also shows results obtained in a medical imaging platform. PMID- 19129021 TI - An activity-subspace approach for estimating the integrated input function and relative distribution volume in PET parametric imaging. AB - Dynamic positron emission tomography (PET) imaging technique enables the measurement of neuroreceptor distributions corresponding to anatomic structures, and thus, allows image-wide quantification of physiological and biochemical parameters. Accurate quantification of the concentration of neuroreceptor has been the objective of many research efforts. Compartment modeling is the most widely used approach for receptor binding studies. However, current compartment model-based methods often either require intrusive collection of accurate arterial blood measurements as the input function, or assume the existence of a reference region. To obviate the need for the input function or a reference region, in this paper, we propose to estimate the input function. We propose a novel concept of activity subspace, and estimate the input function by the analysis of the intersection of the activity subspaces. Then, the input function and the distribution volume (DV) parameter are refined and estimated iteratively. Thus, the underlying parametric image of the total DV is obtained. The proposed method is compared with a blind estimation method, iterative quadratic maximum likelihood (IQML) via simulation, and the proposed method outperforms IQML. The proposed method is also evaluated in a brain PET dataset. PMID- 19129022 TI - Support vector machines for automated recognition of obstructive sleep apnea syndrome from ECG recordings. AB - Obstructive sleep apnea syndrome (OSAS) is associated with cardiovascular morbidity as well as excessive daytime sleepiness and poor quality of life. In this study, we apply a machine learning technique [support vector machines (SVMs)] for automated recognition of OSAS types from their nocturnal ECG recordings. A total of 125 sets of nocturnal ECG recordings acquired from normal subjects (OSAS - ) and subjects with OSAS (OSAS +), each of approximately 8 h in duration, were analyzed. Features extracted from successive wavelet coefficient levels after wavelet decomposition of signals due to heart rate variability (HRV) from RR intervals and ECG-derived respiration (EDR) from R waves of QRS amplitudes were used as inputs to the SVMs to recognize OSAS +/- subjects. Using leave-one-out technique, the maximum accuracy of classification for 83 training sets was found to be 100% for SVMs using a subset of selected combination of HRV and EDR features. Independent test results on 42 subjects showed that it correctly recognized 24 out of 26 OSAS + subjects and 15 out of 16 OSAS - subjects (accuracy = 92.85%; Cohen's kappa value of 0.85). For estimating the relative severity of OSAS, the posterior probabilities of SVM outputs were calculated and compared with respective apnea/hypopnea index. These results suggest superior performance of SVMs in OSAS recognition supported by wavelet based features of ECG. The results demonstrate considerable potential in applying SVMs in an ECG-based screening device that can aid a sleep specialist in the initial assessment of patients with suspected OSAS. PMID- 19129023 TI - Extraction of sources of tremor in hand movements of patients with movement disorders. AB - This paper proposes an efficient method to acquire sources of tremor in patients with movement disorders based on blind source separation of convolutive mixtures. The extracted sources indicated neural activities that might be generated in the central nervous system. Four patients with essential tremor were tested in a set of movement tasks. Subjects wore a data glove that measured finger movements of the hand. The experimental data were then fed to a convolutive-mixture model, which revealed sources that imbibed in them the tremor frequency components of 2- 8 Hz. Time--frequency analysis of these sources might be of potential help to clinicians to devise tasks that can manifest visible tremor from patients. PMID- 19129024 TI - Fetal weight estimation using the evolutionary fuzzy support vector regression for low-birth-weight fetuses. AB - Accurate estimation of fetal weight before delivery is of great benefit to limit the potential complication associated with the low-birth-weight infants. Although the regression analysis has been used as a daily clinical means to estimate the fetal weight on the basis of ultrasound measurements, it still lacks enough accuracy for low-birth-weight fetuses. The ineffectiveness is mainly due to the large inter- or intraobserver variability in measurements and the inappropriateness of the regression analysis. A novel method based on the support vector regression (SVR) is proposed to improve the weight estimation accuracy for fetuses of less than 2500 g. Here, fuzzy logic is introduced into SVR (termed FSVR) to limit the contribution of inaccurate training data to the model establishment, and thus, to enhance the robustness of FSVR to noisy data. To guarantee the generalization performance of the FSVR model, the nondominated sorting genetic algorithm (NSGA) is utilized to obtain the optimal parameters for the FSVR, which is referred to as the evolutionary fuzzy support vector regression (EFSVR) model. Compared with regression formulas, back-propagation neural network, and SVR, EFSVR achieves the lowest mean absolute percent error (6.6%) and the highest correlation coefficient (0.902) between the estimated fetal weight and the actual birth weight. The EFSVR model produces significant improvement (1.9%-4.2%) on the accuracy of fetal weight estimation over several widely used formulas. Experiments show the potential of EFSVR in clinical prenatal care. PMID- 19129025 TI - MicroRNAs and cancer-the search begins! AB - For almost three decades, cancer was thought to result from changes in the structure and/or expression of protein coding genes. The discovery of thousands of genes that produce noncoding RNA (ncRNA) transcripts in the past few years suggested that the molecular biology of cancer is much more complex. MicroRNAs (miRNAs), an important group of ncRNAs, have recently been associated with tumorigenesis by acting either as tumor suppressors or oncogenes. Experimental prediction of miRNA genes is a slow process, because of the difficulties of cloning ncRNAs. Complementary to experimental approaches, a number of computational tools trained to recognize features of the biogenesis of miRNAs have significantly aided in the prediction of new miRNA candidates. By narrowing down the search space, computational approaches provide valuable clues as to which are the dominant features that characterize these regulatory units and which genes are their most likely targets. Moreover, through the use of high throughput expression profiling methods, many molecular signatures of miRNA deregulation in human tumors have emerged. In this review, we present an overview of existing computational methods for identifying miRNA genes and assessing their expression levels, and analyze the contribution of such tools toward illuminating the role of miRNAs in cancer. PMID- 19129026 TI - Two-Phase chief complaint mapping to the UMLS metathesaurus in Korean electronic medical records. AB - The task of automatically determining the concepts referred to in chief complaint (CC) data from electronic medical records (EMRs) is an essential component of many EMR applications aimed at biosurveillance for disease outbreaks. Previous approaches that have been used for this concept mapping have mainly relied on term-level matching, whereby the medical terms in the raw text and their synonyms are matched with concepts in a terminology database. These previous approaches, however, have shortcomings that limit their efficacy in CC concept mapping, where the concepts for CC data are often represented by associative terms rather than by synonyms. Therefore, herein we propose a concept mapping scheme based on a two phase matching approach, especially for application to Korean CCs, which uses term-level complete matching in the first phase and concept-level matching based on concept learning in the second phase. The proposed concept-level matching suggests the method to learn all the terms (associative terms as well as synonyms) that represent the concept and predict the most probable concept for a CC based on the learned terms. Experiments on 1204 CCs extracted from 15,618 discharge summaries of Korean EMRs showed that the proposed method gave significantly improved F-measure values compared to the baseline system, with improvements of up to 73.57%. PMID- 19129027 TI - Pervasive access to MRI bias artifact suppression service on a grid. AB - Bias artifact corrupts MRIs in such a way that the image is afflicted by illumination variations. Some of the authors proposed the exponential entropy driven homomorphic unsharp masking ( E(2)D-HUM) algorithm that corrects this artifact without any a priori hypothesis about the tissues or the MRI modality. Moreover, E(2)D-HUM does not care about the body part under examination and does not require any particular training task. People who want to use this algorithm, which is Matlab-based, have to set their own computers in order to execute it. Furthermore, they have to be Matlab-skilled to exploit all the features of the algorithm. In this paper, we propose to make such algorithm available as a service on a grid infrastructure, so that people can use it almost from everywhere, in a pervasive fashion, by means of a suitable user interface running on smartphones. The proposed solution allows physicians to use the E(2)D-HUM algorithm (or any other kind of algorithm, given that it is available as a service on the grid), being it remotely executed somewhere in the grid, and the results are sent back to the user's device. This way, physicians do not need to be aware of how to use Matlab to process their images. The pervasive service provision for medical image enhancement is presented, along with some experimental results obtained using smartphones connected to an existing Globus based grid infrastructure. PMID- 19129028 TI - An Improved Level Set for Liver Segmentation and Perfusion Analysis in MRIs. AB - Determining liver segmentation accurately from MRIs is the primary and crucial step for any automated liver perfusion analysis, which provides important information about the blood supply to the liver. Although implicit contour extraction methods, such as level set methods (LSMs) and active contours, are often used to segment livers, the results are not always satisfactory due to the presence of artifacts and low-gradient response on the liver boundary. In this paper, we propose a multiple-initialization, multiple-step LSM to overcome the leakage and over-segmentation problems. The multiple-initialization curves are first evolved separately using the fast marching methods and LSMs, which are then combined with a convex hull algorithm to obtain a rough liver contour. Finally, the contour is evolved again using global level set smoothing to determine a precise liver boundary. Experimental results on 12 abdominal MRI series showed that the proposed approach obtained better liver segmentation results, so that a refined liver perfusion curve without respiration affection can be obtained by using a modified chamfer matching algorithm and the perfusion curve is evaluated by radiologists. PMID- 19129029 TI - Multivariate analysis of full-term neonatal polysomnographic data. AB - INTRODUCTION: Polysomnography (PSG) is one of the most important noninvasive methods for studying maturation of the child brain. Sleep in infants is significantly different from sleep in adults. This paper addresses the problem of computer analysis of neonatal polygraphic signals. METHODS: We applied methods designed for differentiating three important neonatal behavioral states: quiet sleep, active sleep, and wakefulness. The proportion of these states is a significant indicator of the maturity of the newborn brain in clinical practice. In this study, we used data provided by the Institute for Care of Mother and Child, Prague (12 newborn infants of similar postconceptional age). The data were scored by an experienced physician to four states (wake, quiet sleep, active sleep, movement artifact). For accurate classification, it was necessary to determine the most informative features. We used a method based on power spectral density (PSD) applied to each EEG channel. We also used features derived from electrooculogram (EOG), electromyogram (EMG), ECG, and respiration [pneumogram (PNG)] signals. The most informative feature was the measure of regularity of respiration from the PNG signal. We designed an algorithm for interpreting these characteristics. This algorithm was based on Markov models. RESULTS: The results of automatic detection of sleep states were compared to the "sleep profiles" determined visually. We evaluated both the success rate and the true positive rate of the classification, and statistically significant agreement of the two scorings was found. Two variants, for learning and for testing, were applied, namely learning from the data of all 12 newborns and tenfold cross-validation, and learning from the data of 11 newborns and testing on the data from the 12th newborn. We utilized information obtained from several biological signals (EEG, ECG, PNG, EMG, EOG) for our final classification. We reached the final success rate of 82.5%. The true positive rate was 81.8% and the false positive rate was 6.1%. DISCUSSION: The most important step in the whole process is feature extraction and feature selection. In this process, we used visualization as an additional tool that helped us to decide which features to select. Proper selection of features may significantly influence the success rate of the classification. We made a visual comparison of the computed features with the manual scoring provided by the expert. A hidden Markov model was used for classification. The advantage of this model is that it determines the future behavior of the process by its present state. In this way, it preserves information about temporal development. PMID- 19129031 TI - WADA service: an extension of DICOM WADO service. AB - The Web Access to Digital Imaging and Communication in Medicine (DICOM) Persistent Objects (WADO) service is standardized as the Web extension to DICOM. This paper analyzes the operational specifications of the WADO service and extends its access capability to the whole DICOM hierarchy (patient, study, series, and object). The proposed Web Access to DICOM Archives (WADA) service, as opposed to WADO, also includes an extrainternal query mechanism and support of medical reports submission. A pilot implementation of WADA as software components and their integration into a three-tier architecture are also presented. Advanced security mechanisms are augmented to ensure communication encryption, user identification, and access restriction to data according to user roles. The proposed service is a simple approach, and can be embedded in any system managing medical images and reports. WADA can also be integrated into the Cross-Enterprise Document Sharing-Imaging (XDS-I) standard, which is considered to be the most likely future standard for medical imaging exchange. PMID- 19129032 TI - Autoregressive-model-based missing value estimation for DNA microarray time series data. AB - Missing value estimation is important in DNA microarray data analysis. A number of algorithms have been developed to solve this problem, but they have several limitations. Most existing algorithms are not able to deal with the situation where a particular time point (column) of the data is missing entirely. In this paper, we present an autoregressive-model-based missing value estimation method (ARLSimpute) that takes into account the dynamic property of microarray temporal data and the local similarity structures in the data. ARLSimpute is especially effective for the situation where a particular time point contains many missing values or where the entire time point is missing. Experiment results suggest that our proposed algorithm is an accurate missing value estimator in comparison with other imputation methods on simulated as well as real microarray time series datasets. PMID- 19129030 TI - Development and preliminary validation of heart rate and breathing rate detection using a passive, ballistocardiography-based sleep monitoring system. AB - Techniques such as ballistocardiography (BCG) that can provide noninvasive long term physiological monitoring have gained interest due to a growing recognition of adverse effects from poor sleep and sleep disorders. The noninvasive analysis of physiological signals (NAPS) system is a BCG-based monitoring system developed to measure heart rate, breathing rate, and musculoskeletal movement that shows promise as a general sleep analysis tool. Overnight sleep studies were conducted on 40 healthy subjects during a clinical trial at the University of Virginia. The NAPS system's measures of heart rate and breathing rate were compared to ECG, pulse oximetry, and respiratory inductance plethysmography (RIP). The subjects were split into a training dataset and a validation dataset, maintaining similar demographics in each set. The NAPS system accurately detected heart rate, averaged over the prescribed 30-s epochs, to within less than 2.72 beats per minute of ECG, and accurately detected breathing rate, averaged over the same epochs, to within 2.10 breaths per minute of RIP bands used in polysomnography. PMID- 19129033 TI - A correlation-test-based validation procedure for identified neural networks. AB - In this study, an enhanced correlation-test-based validation procedure is developed to check the quality of identified neural networks in modeling of nonlinear systems. The new computation algorithm up grades the validation power by including a direct correlation test between residuals and delayed outputs that have been quoted indirectly in the most previous approaches. Furthermore, based on the new validation procedure, three guidelines are proposed in this study to help explain the validation results and the statistic properties of the residuals. It is hoped that this study could promote awareness of why the correlation tests are an effective method of validating identified neural networks, and provide examples how to use the tests in user applications. PMID- 19129034 TI - Computational capabilities of graph neural networks. AB - In this paper, we will consider the approximation properties of a recently introduced neural network model called graph neural network (GNN), which can be used to process-structured data inputs, e.g., acyclic graphs, cyclic graphs, and directed or undirected graphs. This class of neural networks implements a function tau(G,n) is an element of IR(m) that maps a graph G and one of its nodes n onto an m-dimensional Euclidean space. We characterize the functions that can be approximated by GNNs, in probability, up to any prescribed degree of precision. This set contains the maps that satisfy a property called preservation of the unfolding equivalence, and includes most of the practically useful functions on graphs; the only known exception is when the input graph contains particular patterns of symmetries when unfolding equivalence may not be preserved. The result can be considered an extension of the universal approximation property established for the classic feedforward neural networks (FNNs). Some experimental examples are used to show the computational capabilities of the proposed model. PMID- 19129035 TI - Robust stability of Cohen-Grossberg neural networks via state transmission matrix. AB - This brief is concerned with the global robust exponential stability of a class of interval Cohen-Grossberg neural networks with both multiple time-varying delays and continuously distributed delays. Some new sufficient robust stability conditions are established in the form of state transmission matrix, which are different from the existing ones. Furthermore, a sufficient condition is also established to guarantee the global stability for this class of Cohen-Grossberg neural networks without uncertainties. Three examples are used to show the effectiveness of the obtained results. PMID- 19129036 TI - Analysis of continuous attractors for 2-D linear threshold neural networks. AB - This brief investigates continuous attractors of the well-developed model in visual cortex, i.e., the linear threshold (LT) neural networks, based on a parameterized 2-D model. On the basis of existing results on nondegenerate equilibria in mathematics, we further discuss degenerate equilibria for such networks and present properties and distributions of the equilibria, which enables us to draw the coexistence conditions of nondegenerate and degenerate equilibria (e.g., singular lines). Our theoretical results provide a useful framework for precise tuning on the network parameters, e.g., the feedbacks and visual inputs. Simulations are also presented to illustrate the theoretical findings. PMID- 19129037 TI - Lattice dynamical wavelet neural networks implemented using particle swarm optimization for spatio-temporal system identification. AB - In this brief, by combining an efficient wavelet representation with a coupled map lattice model, a new family of adaptive wavelet neural networks, called lattice dynamical wavelet neural networks (LDWNNs), is introduced for spatio temporal system identification. A new orthogonal projection pursuit (OPP) method, coupled with a particle swarm optimization (PSO) algorithm, is proposed for augmenting the proposed network. A novel two-stage hybrid training scheme is developed for constructing a parsimonious network model. In the first stage, by applying the OPP algorithm, significant wavelet neurons are adaptively and successively recruited into the network, where adjustable parameters of the associated wavelet neurons are optimized using a particle swarm optimizer. The resultant network model, obtained in the first stage, however, may be redundant. In the second stage, an orthogonal least squares algorithm is then applied to refine and improve the initially trained network by removing redundant wavelet neurons from the network. An example for a real spatio-temporal system identification problem is presented to demonstrate the performance of the proposed new modeling framework. PMID- 19129038 TI - Current advanced statistical issues in clinical research---adaptive designs and translational medicine. PMID- 19129039 TI - A regulatory view of adaptive trial design. AB - Developing a new medicine is an expensive and time-consuming process. Researchers are interested in applying better designs to expedite the approval of potential medicinal products. Adaptive designs, which allow for some types of prospectively planned mid-study change, can improve the efficiency of a trial and maximize the chance of success. Possible design adaptations of clinical trials include sample size re-estimation, change in primary endpoint, interim dropping of treatment arms, change in statistical hypothesis, and change in the primary analysis. In this article, the regulatory considerations of the methodological issues with respect to adaptive design are discussed. Several examples of design adaptation that the Center for Drug Evaluation has encountered during the past 3 years are presented. PMID- 19129040 TI - The significance of adaptive design in r&d in Japan. AB - In the past decade, the number of drug and biological product submissions to the United States Food and Drug Administration has been slowly declining, while biomedical research spending has increased. A similar situation can also be seen in Japan: the number of domestic clinical trials is diminishing, and the cost of conducting a trial is rising. To prevent further decreases in the number of clinical trials, there is a need for an innovative strategy such as an adaptive design in research and development. Integrative celerity research aims to combine critical path and translational research, and seek update and participation in global clinical research. Participation in global studies through an adaptive design raises statistical concerns, which can be dealt with by adapting bridging studies. As a result of the restricted number of patients before approval in the adaptive design, safety issues must be guaranteed. Thus, establishing an effective and strong safety network between medical facilities is crucial. Japans mission is to develop better drugs more efficiently and to investigate new drug methodologies for participation in global/Asian studies. Team work between clinical trial specialists, computer scientists, medical doctors, and statisticians is important for the success of both adaptive design and construction of a safety network between medical facilities in Japanese research and development. PMID- 19129041 TI - Considerations in adapting clinical trial design. AB - The concept of adaptation of trial design during the course of a clinical trial has recently drawn much interest from the pharmaceutical industry. The interest arises partly because statistical decision trees employed to address multiple complex clinical hypotheses within a clinical trial are increasingly complex, and the statistical information generated from learning data prior to designing the trial is often insufficient to provide informative guidance for planning a pivotal trial. While the conventional fixed designs, which usually permit no modification influenced by the internal trial data of key design specifications, often cannot cover the range of complex statistical decision trees that must be prespecified in the study protocol, it seems natural to consider modifications of trial design at some point in the trial. In regulatory practice, some adjustments to study protocols are mostly made known to regulatory agencies in the form of so called protocol amendments. However, such design modifications may demand careful consideration in dealing with any biases that may be caused by the adaptation, and may impede the interpretability of trial results. PMID- 19129042 TI - Utility of adaptive strategy and adaptive design for biomarker-facilitated patient selection in pharmacogenomic or pharmacogenetic clinical development program. AB - In the early to late phases of conventional clinical trials, improvement of disease status at study baseline is the anchor of an effective treatment measured by therapeutic response. These population-based clinical trials do not formally account for disease-associated marker genotype or genome-associated therapeutic response. We discuss alternative study designs in pharmacogenomic or pharmacogenetic clinical trials for genomic or genetic biomarker development, and for formally assessing the clinical utility of genomic or genetic (composite) biomarkers. A two-stage adaptive strategy from completed, ongoing or prospectively planned pharmacogenomic or pharmacogenetic clinical trials is described for development of a genomic or genetic biomarker. We present two types of adaptive design: (1) the genomic biomarker is developed external to the clinical trial, which is designed for treatment effect inference; and (2) first stage data are used to explore a genomic biomarker, but statistical inference of treatment effect in the genomically or genetically defined biomarker subset is only performed at the second stage of the same trial. When the null hypothesis of no treatment effect in all randomized patients and the genomic patient subset are prospectively specified, we compare the statistical power between fixed and adaptive designs. We also compare the two types of adaptive design. Results from simulation studies showed that adaptive design is more powerful than fixed design for those genomic or genetic biomarkers whose clinical utility is predictive of treatment effect. Pursuit of adaptive design gains at least 20% to more than 30% genomic patient subset power when the genomic biomarker status is readily usable at study initiation, in comparison to when it is explored using the first-stage data of the same clinical trial. In exploratory studies, adaptive strategy provides wide flexibility in the process of genomic or genetic biomarker development. In contrast, an adaptive design trial that employs limited flexibility, and is an adequate and well-controlled investigation, has a greater power gain than a fixed design trial, in which the genomic biomarker is capable of predicting treatment effects that pertain only to the prespecified genomic or genetic patient subset. PMID- 19129043 TI - Statistical evaluation of quality performance on genomic composite biomarker classifiers. AB - BACKGROUND/PURPOSE: After completion of the Human Genome Project, genomic composite biomarker classifiers (GCBCs) became available. However, quality performance of GCBCs varies. We propose statistical methods for evaluation of the quality performance of GCBCs on selection of differentially expressed genes, agreement and reproducibility. METHODS: For detection of differentially expressed genes, an interval hypothesis was employed to take into account both biological and statistical significance. The concordance correlation coefficient (CCC) was used to evaluate the agreement of expression levels of technical replicates. The intraclass correlation coefficient (ICC) was suggested to assess the reproducibility between laboratories. RESULTS: A two one-sided test procedure was proposed to test the interval hypothesis. Statistical methods based on the generalized pivotal quantities for CCC and ICC were suggested to test the hypotheses for agreement and reproducibility. Simulation results demonstrated that all three methods could adequately control the type I error rate at the nominal level for assessment of differentially expressed genes, agreement and reproducibility. CONCLUSION: Three appropriate statistical methods were developed for evaluation of quality performance on differentially expressed genes, agreement and reproducibility of GCBCs. PMID- 19129044 TI - Statistical methods for targeted clinical trials under enrichment design. AB - BACKGROUND/PURPOSE: After completion of the Human Genome Project, disease targets at the molecular level can be identified. Treatment for these specific targets can be developed with the individualized treatment of patients becoming a reality. However, the accuracy of diagnostic devices for molecular targets is not perfect and statistical inference for treatment effects of the targeted therapy is biased. We developed statistical methods for an unbiased inference for the targeted therapy in patients who truly have the molecular targets. METHODS: Under the enrichment design, for binary data, we propose using the expectation maximization (EM) algorithm with the bootstrap method, to incorporate the inaccuracy of the diagnostic device for detection of the molecular targets for inference of the treatment effects. A simulation study was conducted to empirically investigate the performance of the proposed estimation and testing procedures. A numerical example illustrates the application of the proposed method. RESULTS: Simulation results demonstrated that the proposed estimation method was unbiased, with adequate precision, and the confidence interval provided satisfactory coverage probability. The proposed testing procedure adequately controlled the size with sufficient power. The numerical example showed that a statistically significant treatment effect could be obtained when the inaccuracy of the diagnostic device was taken into account. CONCLUSION: Our proposed estimation and testing procedures are adequate statistical methods for the inference of the treatment effect for patients who truly have the molecular targets. PMID- 19129045 TI - Statistical tests for one-way/two-way translation in translational medicine. AB - Translational medicine has been defined as bench-to-bedside research, where a basic laboratory discovery becomes applicable to the diagnosis, treatment or prevention of a specific disease, and is brought forth by either a physician/scientist who works at the interface between the research laboratory and patient care, or by a team of basic and clinical science investigators. Statistics plays an important role in translational medicine to ensure that the translational process is accurate and reliable, with statistical assurance. For this purpose, statistical criteria for assessment of one-way and two-way translation are proposed. Under a well established and validated translational model, statistical tests for one-way and two-way translation are discussed. Some discussion on lost in translation is also given. PMID- 19129046 TI - On Two-stage Seamless Adaptive Design in Clinical Trials. AB - In recent years, the use of adaptive design methods in clinical research and development based on accrued data has become very popular because of its efficiency and flexibility in modifying trial and/or statistical procedures of ongoing clinical trials. One of the most commonly considered adaptive designs is probably a two-stage seamless adaptive trial design that combines two separate studies into one single study. In many cases, study endpoints considered in a two stage seamless adaptive design may be similar but different (e.g. a biomarker versus a regular clinical endpoint or the same study endpoint with different treatment durations). In this case, it is important to determine how the data collected from both stages should be combined for the final analysis. It is also of interest to know how the sample size calculation/allocation should be done for achieving the study objectives originally set for the two stages (separate studies). In this article, formulas for sample size calculation/allocation are derived for cases in which the study endpoints are continuous, discrete (e.g. binary responses), and contain time-to-event data assuming that there is a well established relationship between the study endpoints at different stages, and that the study objectives at different stages are the same. In cases in which the study objectives at different stages are different (e.g. dose finding at the first stage and efficacy confirmation at the second stage) and when there is a shift in patient population caused by protocol amendments, the derived test statistics and formulas for sample size calculation and allocation are necessarily modified for controlling the overall type I error at the prespecified level. PMID- 19129047 TI - Statistical methods in translational medicine. AB - This study focuses on strategies and statistical considerations for assessment of translation in language (e.g. translation of case report forms in multinational clinical trials), information (e.g. translation of basic discoveries to the clinic) and technology (e.g. translation of Chinese diagnostic techniques to well established clinical study endpoints) in pharmaceutical/clinical research and development. However, most of our efforts will be directed to statistical considerations for translation in information. Translational medicine has been defined as bench-to-bedside research, where a basic laboratory discovery becomes applicable to the diagnosis, treatment or prevention of a specific disease, and is brought forth by either a physicianscientist who works at the interface between the research laboratory and patient care, or by a team of basic and clinical science investigators. Statistics plays an important role in translational medicine to ensure that the translational process is accurate and reliable with certain statistical assurance. Statistical inference for the applicability of an animal model to a human model is also discussed. Strategies for selection of clinical study endpoints (e.g. absolute changes, relative changes, or responder-defined, based on either absolute or relative change) are reviewed. PMID- 19129048 TI - Translation in different diagnostic procedures---traditional Chinese medicine and Western medicine. AB - Recently, the modernization of traditional Chinese medicines (TCM) for treatment of patients with critical and/or life-threatening diseases has attracted much attention in the pharmaceutical industry. However, there exist essential differences in the evaluation of the efficacy and safety of a TCM as compared with a typical Western medicine (WM), even though they are for the same indication. Therefore, the modernization of a TCM should be based on a scientific evaluation of the safety and effectiveness of the TCM in terms of well established quantitative criteria. We propose a study design to study the calibration and validation of the Chinese diagnostic procedure for evaluation of a TCM, with respect to a well-established clinical endpoint for evaluation of a WM. Statistical validation of such an instrument is essential to have an accurate and reliable clinical assessment of the performance of the TCM. Similar to the validation of a typical quality of life instrument, some validation performance characteristics such as validity, reliability, and ruggedness are considered. In this article, a design for validation of a standard quantitative instrument to be commonly employed for diagnosis of patient function/activity, performance, disease signs and symptoms, and disease status and severity based on Chinese diagnostic practice is proposed. Methods for statistical validation of the standard instrument are derived. More specifically, for validation of the TCM diagnostic instrument, we consider the following validation performance characteristics (parameters): validity (or accuracy), reliability (or precision), and ruggedness (interrater variability). A numerical example is given to illustrate the proposed methods for validation of the Chinese diagnostic procedure. PMID- 19129049 TI - HIV-1 at age 25: some thoughts for Taiwan and China. PMID- 19129050 TI - Waking up the sleepers: HIV latency and reactivation. AB - In a patient infected with HIV-1, the presence of latently infected cells from which the virus can be reactivated and rekindle HIV infection in the patient necessitates lifelong administration of antiretroviral treatment. The biology of HIV latency and viral silencing is now becoming clearer at a molecular and cellular level. However, our understanding of HIV-1 latency in vivo is still inadequate. Attempts to therapeutically reactivate the virus in infected patients have yielded disappointing results. This article reviews the research and clinical findings and discusses current thinking on the subject of HIV latency and reactivation. PMID- 19129051 TI - Cognitive development at age 8 years in very low birth weight children in Taiwan. AB - BACKGROUND/PURPOSE: All children should have some developmental screening periodically throughout childhood, especially those who were born prematurely. There is limited information about the development of children with very low birth weight (VLBW; birth weight < 1500 g) beyond the preschool age in Taiwan. We evaluated intelligence quotient (IQ) and cognitive ability of prematurely born school-aged children in Taiwan. METHODS: This was a multicenter study of VLBW and full-term children born between 1995 and 1997 at four hospitals in northern Taiwan. We used the Wechsler Intelligence Scale for Children, 3rd Edition (WISC III), to assess these children. Demographic data were recorded including maternal and paternal age, education, birth weight, gestational age, and gender. RESULTS: A total of 189 children (130 with VLBW born prematurely and 59 born at full term) were recruited. There were significant differences in performance IQ (PIQ; 90.16 +/- 17.05 vs. 108.51 +/- 15.65, p < 0.001), verbal IQ (VIQ; 97.43 +/- 15.62 vs. 111.78 +/- 13.65, p < 0.001), full-scale IQ (FSIQ; 93.14 +/- 16.33 vs. 111.05 +/- 14.81, p < 0.001), verbal comprehension index score (VCIS; 98.06 +/- 15.53 vs. 112.47 +/- 13.74, p < 0.001), perceptional organization index score (POIS; 92.39 +/- 17.13 vs. 109.42 +/- 14.87, p < 0.001) and freedom from distractibility index score (FDIS; 98.34 +/- 17.71 vs. 110.53 +/- 10.94, p = 0.008). There was no correlation between perinatal outcomes and FSIQ. CONCLUSION: Our results suggest that VLBW children have significantly lower PIQ, VIQ, FSIQ, VCIS, POIS and FDIS at primary school compared with full-term children. PMID- 19129052 TI - Non-psychiatric physicians' knowledge, attitudes and behavior toward depression. AB - BACKGROUND/PURPOSE: Depression is a major health concern, often treated by non psychiatrists. This study assessed self-reported knowledge, attitudes and treatment practices of non-psychiatric physicians in the recognition and management of depression. METHODS: Survey questionnaires were given to non psychiatric physicians who attended a depression training program. We asked physicians about their current clinical practice, knowledge, confidence, attitudes and perceived barriers to care regarding recognition and management of patients with depression. RESULTS: Of 524 eligible non-psychiatric physicians, 375 (72%) completed surveys. The majority of physicians held a strong sense of responsibility for managing depression, although they provided treatment to only a small proportion of depressed patients. Most of them were not confident treating depressed patients, and they reported that incomplete knowledge and training were major barriers that limited their involvement. The patient and organization barriers were not related to reported management, but the physician barriers (lack of skills and knowledge in managing depression) were related to reported rate of treatment. Age, prior depression training, and education were major contributing factors to domains of knowledge, attitude and behavior, in terms of the number of domains involved. Family physician orientation was associated with higher score on knowledge scale, but not with other variables of attitude and behavior. CONCLUSION: Our study suggests that non-psychiatrists may also play a role in the care of depression, but identifying and managing depression can be a challenge to them. Attitudinal barriers, confidence, and knowledge of treatment may compromise the physicians ability to manage depression. Future interventions and educational efforts need to address each of these issues. PMID- 19129053 TI - Changing pattern of ectopic pancreas: 22 years of experience in a medical center. AB - BACKGROUND/PURPOSE: Ectopic pancreas is usually a silent gastrointestinal malformation. It may become clinically evident when complicated by chronic inflammation or by growth. More ileal ectopic pancreas has been found in recent years in our hospital. We report the clinical manifestation of ectopic pancreas over the past 22 years. METHODS: We reviewed the medical records of patients seen between May 1984 and December 2005 at Mackay Memorial Hospital, with a diagnosis of ectopic pancreas, and extracted clinical and histopathology data from the records. RESULTS: A total of 39 patients (18 male, 21 female; mean age, 46 years) were diagnosed with ectopic pancreas. Most patients were aged between 30 and 50 years. Only 15 (38%) had symptoms suggestive of ectopic pancreas. These included abdominal pain (n = 9), upper gastrointestinal bleeding (n = 5), and abdominal distension (n = 2). The diagnosis in the other 24 patients was made incidentally, usually during surgery for other conditions. While lesions in the stomach were more likely to be diagnosed because of symptoms (12 of 13), lesions in the small bowel were almost always diagnosed incidentally. Only one of eight in the duodenum, one of 10 in the jejunum, and one of eight in the ileum, were isolated findings. One case of ectopic pancreas was detected by capsule endoscopy. CONCLUSION: Ectopic pancreas can be found in various parts of the gastrointestinal tract. The high proportion of ileal ectopic pancreas is unexpected and needs further study. PMID- 19129054 TI - Mass screening of suspected febrile patients with remote-sensing infrared thermography: alarm temperature and optimal distance. AB - BACKGROUND/PURPOSE: Detection of fever has become an essential step in identifying patients who may have severe acute respiratory syndrome (SARS) or avian influenza. This study evaluated infrared thermography (IRT) and compared the influence of different imagers, ambient temperature discrepancy, and the distance between the subject and imager. METHODS: IRT-digital infrared thermal imaging (IRT-DITI), thermoguard, and ear drum IRT were used for visitors to Municipal Wang Fang Hospital, Taipei, Taiwan. The McNemar and Chi-squared test, standard Pearson correlation, ANOVA, intraclass correlation coefficient (ICC), and receiver operating characteristic curve (ROC) analysis were used to calculate the alarm temperature for each imager. RESULTS: A total of 1032 subjects were recruited. Different distances and ambient temperature discrepancy had a significant influence on thermoguard, and lateral and frontal view DITI. By ICC analysis, a significant difference was found at 10 m distance between ear drum IRT and thermoguard (r = 0.45), lateral view DITI (r = 0.37), and frontal view DITI (r = 0.44). With ROC analysis, the optimal preset cut-off temperatures for the different imagers were: 36.05 degrees C for thermoguard (area under the curve [AUC], 0.716), 36.25 degrees C for lateral view DITI (AUC, 0.801), and 36.25 degrees C for frontal view DITI (AUC, 0.812). CONCLUSION: The temperature readings obtained by IRT may be used as a proxy for core temperature. An effective IRT system with a strict operating protocol can be rapidly implemented at the entrance of a hospital during SARS or avian influenza epidemics. PMID- 19129055 TI - A 6-year retrospective epidemiologic study of pediatric pneumococcal pneumonia in Taiwan. AB - BACKGROUND/PURPOSE: Community-acquired pneumonia (CAP) is one of the most common diseases during childhood, and its features vary between countries. Death from CAP is rare in industrialized countries, but it is the leading cause of mortality in developing countries. The purpose of this study was to determine the epidemiology and morbidity of pneumococcal CAP (PCAP) in children hospitalized with pneumonia in Taiwan. METHODS: We retrospectively screened the medical records of 12,914 children hospitalized with pneumonia at four hospitals in Taiwan from 2000 to 2005. This included a suspected PCAP group (2350 patients) and a group with likely/definite pneumococcal pneumonia (PP group, 901 patients). RESULTS: From 2000, the frequency of suspected PCAP increased gradually every year, and peaked in 2003 (22.14%); while the proportion of likely/definite PP was highest in 2004 (9.75%). The likely/definite PP group accounted for 6.98% of the hospitalized pneumonia patients from 2000 to 2005. Of children hospitalized with pneumonia, 17.9% of the suspected PCAP group and 21.3% of the PP group were admitted to the intensive care unit (ICU). Twenty-three patients died (14 in the PCAP and 9 in the PP groups) and nine (5 in the PCAP and 4 in the PP groups) were discharged with sequelae. CONCLUSION: Our results indicate that children hospitalized with PCAP are common in Taiwan. About 20% of these children need ICU admissions, and some still have a poor outcome. Effective immunization programs with pneumococcal conjugate vaccine are needed for disease control and prevention. PMID- 19129056 TI - Association between high myopia and progression of visual field loss in primary open-angle glaucoma. AB - BACKGROUND/PURPOSE: Taiwan has a very high prevalence rate of myopia. We retrospectively studied the influence of myopia on the progression of visual field (VF) loss in primary open-angle glaucoma (POAG) patients. METHODS: We studied 515 POAG patients for a minimum follow-up period of 5 years. VF examination was performed with Humphrey perimeter, 30-2 SITA standard program, every 6 months. A point-wise numerical comparison was applied to judge the VF changes. Test points showing more than 1.0 dB of sensitivity loss in mean defect were identified. A location was considered to have progression if it was detected on two consecutive visits. Progression of VF loss was confirmed if three or more test points deteriorated. Multivariate logistic regression was used to evaluate the association between progression of VF loss and various risk factors. RESULTS: There were 262 cases. Progression of VF loss occurred in 57 eyes (21.8%) during the 5-year follow-up period. Logistic regression revealed that the deterioration was associated with older age, higher mean intraocular pressure, larger vertical cup-to-disc ratio, and greater myopic refraction status. The incidence of VF loss progression was 15.1% in the group of eyes with myopia less than -3 D, 10.5% in the group with -3 D to -6 D, 34.4% in the group with -6 D to -9 D, and 38.9% in the group with myopia greater than -9 D. CONCLUSION: POAG patients with myopia greater than -6 D had a greater progression of VF loss. PMID- 19129057 TI - Herpes zoster infection associated with poor peripheral blood hematopoietic stem cell mobilization. AB - The efficacy of peripheral blood hematopoietic stem cell (PBSC) harvest is important for successful autologous transplantation. The impact of viral infection on PBSC mobilization has rarely been reported. Here, we report a patient with relapsed diffuse large B-cell lymphoma who experienced disseminated cutaneous herpes zoster infection during the neutropenic phase of PBSC mobilization. A markedly reduced number of PBSCs was initially harvested (1.72 x 10(6)/kg, 77.2% reduction), followed by a sufficient number (7.55 x 10(6)/kg) during remobilization with the same mobilization regimen when herpes zoster infection had subsided. Because of the temporal association, we suggest that herpes zoster infection is a risk factor for poor PBSC mobilization, and remobilization with the same regimen is feasible. PMID- 19129058 TI - Duodenal metastasis of malignant pleural mesothelioma. AB - Metastatic malignant mesothelioma of the pleura is uncommon at the time of initial diagnosis. The gastrointestinal lumen is rarely found at autopsy in patients with widespread disease. Here, we describe an extremely rare case of isolated duodenal metastasis of sarcomatoid mesothelioma of the pleura in a 73 year-old man, without memory of any direct exposure to asbestos. The possibility of gastrointestinal tract metastasis should be considered in the presence of anemia or positive occult blood test in patients with malignant pleural mesothelioma. PMID- 19129059 TI - X-linked myotubular myopathy with a novel MTM1 mutation in a Taiwanese child. AB - We report a male, preterm newborn infant with X-linked myotubular myopathy, the most severe type of the disease. He presented at birth with generalized hypotonia, difficulty in swallowing, and respiratory distress with frequent episodes of atelectasis. The infant had a long thin face, generalized hypotonia, and arachnodactyly. Diagnosis was based on fetal history, muscle histopathology, electron microscopy and a genetic study. A base pair change was detected in exon 11 of the MTM1 gene: c.1160C>A, which caused an amino acid change, p.S387Y. The father's gene was normal but the mother had the same mutation as her son and was thus a carrier. PMID- 19129060 TI - Surgery, ablation or combination. PMID- 19129062 TI - New and investigational triazole agents for the treatment of invasive fungal infections. AB - The incidence of invasive fungal infections (IFIs) caused by both common and uncommon opportunistic fungi is increasing along with emerging fungal resistance. Since traditional agents (amphotericin B, fluconazole, itraconazole) are limited by an inadequate spectrum of activity, drug resistance or toxicity, there is a great interest in the development of new antifungal agents for treatment of IFIs in high-risk populations. In recent years a number of systemic antifungal drugs have become available and options for treatment of IFIs have expanded. A new generation of triazole agents (voriconazole, posaconazole, isavuconazole, ravuconazole and albaconazole), with a broad spectrum of activity and sufficient improvements in potency to overcome resistance have emerged and represent an alternative to conventional antifungals for the prevention and treatment of IFIs. This article focuses on the microbiology, pharmacology, clinical efficacy and safety of the new antifungal triazole generation. PMID- 19129063 TI - Innate immunity in sepsis pathogenesis and its modulation: new immunomodulatory targets revealed. AB - Sepsis is complex clinical manifestation of an organism's overwhelming and unregulated immune response to infection. Despite more than 25 years of extensive research, sepsis and systemic inflammatory response syndrome (SIRS) remain the major causes of death in hospital intensive care units (ICUs). The mortality rate associated with sepsis varies from 30% to 70%. Up to now no specific treatment for sepsis exists, so in order to lower mortality rates associated with sepsis, it is important to search for better targets for treatment. The understanding of immunopathogenetic mechanisms involved in sepsis development has provided significant advances in the field of innate immunity, which plays an important role in the onset of sepsis and associated mortality. Not only have the pathways involved in sepsis development been defined but also various targets that can be used as an immunomodulatory approach during sepsis treatment. This review explores the role of innate immunity in the development of sepsis and its modulation as a future immunomodulatory approach for sepsis treatment. PMID- 19129064 TI - In vitro combinations of five intravenous antibiotics with dalfopristin quinupristin against Staphylococcus aureus in a 3-dimensional model. AB - We compared the in vitro activity of dalfopristin and quinupristin combined with five intravenous antibiotics in a 3-dimensional model. We tested six strains of Staphylococcus aureus selected with different patterns of resistance to methicillin and erythromycin. Dalfopristin and quinupristin displayed a very synergistic activity against all the strains with a mean 16- or 32-fold decrease of inhibitory concentrations in combination. That synergy was even better against erythromycin-resistant strains. In combination with tigecycline or fosfomycin, the antibacterial activity could be consistently enhanced with the same decrease of inhibitory concentrations. A synergy was also observed, less regularly and at a lower level, with rifampin, gentamicin or vancomycin. Combinations of dalfopristin and quinupristin with tigecycline or fosfomycin could be very interesting in clinical practice because the inhibitory effect could be achieved with very low concentrations of each component, even when erythromycin-resistant strains are concerned. PMID- 19129065 TI - One tube multiplex PCR for simple screening of SCCmec I-V types of methicillin resistant Staphylococcus aureus. AB - In this study, a rapid and simple one-tube multiplex PCR (M-PCR) method was developed for Staphylococcal Cassette Chromosome mec (SCCmec), primary screening. One hundred and eight methicillin-resistant Staphylococcus aureus (MRSA) isolates with SCC mec typing results using the Oliveira and de Lencastre method (Oliveira and de Lencastre. Antimicrob. Agents Chemother. 46:2155-2161, 2002), were evaluated for the efficacy of our method. Typing results were also compared to the recently published PCR protocol by Kondo et al., (Kondo et al., Antimicrob Agents Chemother. 51:264-74, 2007). The one-tube M-PCR method showed comparable efficacy to the other two methods. None of the isolates were classified as non typable in Kondo's and our method. Two formerly assigned SCCmec type I isolates by Oliveira and de Lencastre were determined as SCCmec type IV by Kondo's and the method in this study. In conclusion, the one-tube M-PCR method developed in this study increased the rapidity and simplicity for primary screening SCCmec type I to V MRSA. PMID- 19129066 TI - An update on invasive pneumococcal antibiotic resistance in Turkey, 2008. AB - Cumulative pneumococcal antibiotic resistance profiles in Turkey in 2008, verified from local studies are as follows: penicillin 35%, cefotaxime 2%, ceftriaxone 1%, trimethoprim-sulfamethoxazole 39%, tetracycline 19%, erythromycin 18%, azithromycin 18%, clarithromycin 10%, ofloxacin 13%, clindamycin 9%, chloramphenicol 5%, rifampicin 2%, levofloxacin 2%, moxifloxacin 0%, gemiloxacin 0%, telithromycin 0%, vancomycin 0%. PMID- 19129067 TI - Gram-negative non-fermenting bacteria from food-producing animals are low risk for hospital-acquired infections. AB - The aim of this study was to investigate possible indications of epidemiological relationships between Pseudomonas aeruginosa and Acinetobacter baumannii isolated from food-producing animals and those of clinical origin. Screening for P. aeruginosa and A. baumannii isolates from food-producing animals was carried out on 1381 samples. Susceptibility testing and PCR amplification of resistance genes were determined. Isolate clonal relatedness was established by PFGE. Forty-one P. aeruginosa and 16 A. baumannii were detected. All P. aeruginosa isolates were sensitive to ciprofloxacin, ceftazidime and piperacillin/tazobactam and seven isolates had low-level imipenem resistance. All A. baumannii isolates were sensitive to imipenem, meropenem, ciprofloxacin and piperacillin/tazobactam but were resistant to ceftazidime. The imipenem-resistant P. aeruginosa and ceftazidime-resistant A. baumannii had different PFGE patterns compared to those of human origin. Based on the findings presented here, animal isolates were not multidrug resistant and they do belong to a different pool from those of humans. PMID- 19129068 TI - Klebsiella pneumoniae LO10 producing extended-spectrum beta-lactamase SHV-12 in Saudi Arabia. AB - Klebsiella pneumoniae LO10 was responsible for an outbreak that occurred in the neonatal unit at Security Forces Hospital, Kingdom of Saudi Arabia. Over a period of eight months nine cases of bacteremia resulted in two deaths. Resistance to third generation cephalosporins was transferred from strain LO10 to E. coli by both conjugation and transformation. Sequence determination of the plasmid gene from two transconjugants and one transformant indicated that resistance was carried by a ca.100-kb plasmid encoding beta-lactamase SHV-12. This is the first description of a K. pneumoniae producing a type SHV-12 extended spectrum beta lactamase in Riyadh. Long term exposure to antibiotics, prolonged stay, and heavy use of third generation cephalosporins contributed to the spread of the resistant strain in the unit. Strict infection control measures led to control of the outbreak. PMID- 19129069 TI - Prevalence of type III secretion protein exoenzymes and antimicrobial susceptibility patterns from bloodstream isolates of patients with Pseudomonas aeruginosa bacteremia. AB - The purpose of this study was to determine the prevalence of two type III secretion effector proteins, exoU and exoS from bloodstream isolates of hospitalized patients with Pseudomonas aeruginosa (PSA) bacteremia, to characterize antimicrobial susceptibility patterns, and to compare mortality rates. PSA bloodstream isolates and antibiotic susceptibility profiles were collected from a university-affiliated hospital. ExoS and exoU genes were detected by polymerase chain reaction. Hospital mortality was assessed by medical chart review. 119 of 122 (97.5%) PSA bloodstream isolates contained either the exoS or exoU genes. ExoS was the most prevalent (n=86; 70.5%) followed by exoU (n=31; 25.4%), both genes (n=2; 1.6%) or neither gene (n=3; 2.5%). Isolates containing the exoU gene were significantly more likely to be resistant to cefepime, ceftazidime, piperacillintazobactam, carbapenems, fluoroquinolones, and gentamicin (p<0.05 for all). Mortality was high in patients with PSA bacteremia and did not differ among patients infected with the exoS isolates (n=37; 43%) or exoU isolates (n=11; 35%). One of two type III secretion effector proteins were almost universally present in PSA bloodstream isolates. Isolates containing the exoU gene were more likely to be resistant to multiple antibiotics. PMID- 19129070 TI - Antimicrobial resistance among uropathogens responsible for community-acquired urinary tract infections in an Italian community. AB - This study aimed to establish the pattern of the antimicrobial resistance among the leading uropathogens causing community-acquired UTIs in an area of the region of Apulia, Southern Italy. Twenty-one thousand and two hundred outpatients, 6,893 males and 14,307 females, were enrolled. Urinary isolates were identified by conventional methods and the susceptibility to 18 antimicrobials determined. Recognized uropathogens Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis were isolated from 3175 positive samples, E. coli accounting for 68.04% of positive cultures. Most overall resistance was to ampicillin, while the resistance rate to cephalothin was higher than that of third generation cephalosporins. Although to a different degree, all the bacteria had an overall good susceptibility rate to quinolones as well as to fosfomycin but increased resistance to sulfamethoxazole/trimethoprim. Our results confirm that E. coli is the leading uropathogen and provide information about the antimicrobial susceptibility patterns of the main pathogens causing community-acquired UTIs. These findings should be taken into account to help maintain the safety and efficacy of treatment for community-acquired UTIs. PMID- 19129071 TI - The antitumor activity of meisoindigo against human colorectal cancer HT-29 cells in vitro and in vivo. AB - The study was conducted to examine the antitumor activity of meisoindigo on HT-29 cells in vitro and in vivo. The cytotoxicity of meisoindigo was evaluated by MTT assay. The related genes and proteins were inspected with RT-PCR and western blot assay respectively, and the effects of meisoindigo on the cell cycle were analyzed by flow cytometry. The efficacy of meisoindigo in vivo was evaluated in an HT-29 cell xenograft nude mice model. The results show that meisoindigo effectively inhibits HT-29 cell proliferation (IC(50) 4.3 mmol/L), arrests HT-29 cells in G2/ M phase and induces HT-29 cell apoptosis. The downstream genes and proteins of GSK-3beta(ser(9)) expression level decrease. Meisoindigo significantly inhibits the HT-29 xenograft tumors growth at the dose of 100 mg/kg. The mechanism of meisoindigo activity against HT-29 cells may be related to its inhibition of glycogen synthase kinase-3beta, GSK-3beta(ser(9)) phosphorylation in Wnt signaling pathway. These findings indicate the potential value of meisoindigo for the treatment of colorectal cancer. PMID- 19129072 TI - Role of ABCB1 and ABCC1 gene induction on survival in locally advanced breast cancer. AB - Drug resistance to chemotherapy in patients with locally advanced breast cancer results in a decrease in treatment efficacy and in patient survival. This study aimed to evaluate the impact of ABCB1 and ABCC1 gene induction during chemotherapy on disease-free and overall survival of breast cancer patients. Patients with locally advanced breast cancer were prospectively included. All patients were preoperatively treated with chemotherapy and underwent mastectomy. ABCB1 and ABCC1 gene and protein expressions were evaluated both before and after chemotherapy and investigated as molecular predictive parameters affecting diseasefree and overall survival. ABCB1 and ABCC1 gene expressions were evaluated with RTPCR following RNA isolation from tissue samples. P-glycoprotein and MRP1 in tissues were detected using immunohistochemistry. Twenty-five female patients treated with either doxorubicin or epirubicin were included. Median follow-up time was 36 months during which 11 patients (44%) had recurrence, all of whom died. Mean disease-free survival for patients with and without ABCB1 gene induction was 13 and 55 months (p=0.0004), respectively, whereas overall survival was 21 and 57 months (p=0.0025), respectively. Mean disease-free survival for patients with and without ABCC1 gene induction was 32 and 48 months (p=0.97), respectively, and overall survival was 43 and 49 months (p=0.36), respectively. ABCB1 gene induction decreases disease-free and overall survival in patients with locally advanced breast cancer due to anthracycline resistance. Detecting ABCB1 gene expression during chemotherapy helps to increase the efficacy of drug treatment by choosing the appropriate drugs resulting in prolonged survival. PMID- 19129073 TI - Protective effect of leuprolide on ovarian function in young women treated with adjuvant chemotherapy for early breast cancer: a multicenter phase II study. AB - The aim of this study was to evaluate the protective effect of concomitant leuprolide treatment on ovarian function in young women undergoing adjuvant chemotherapy for early breast cancer. 19 women, median age 36.5 years (range 26 40 years), with operable breast cancer and negative hormonal receptors, received six cycles of FEC 100 regimen as adjuvant chemotherapy and co-treatment with leuprolide. Menstrual resumption was gained in all patients in a median time of 5 months (range 3-8). Follicle-stimulating hormone and estradiol assessment was performed in all patients. The return to pre-menopausal values was achieved within 6 months of the last leuprolide administration. At a median follow-up of 3 years (range 1-5 years), no patient relapsed and four full-term pregnancies were recorded in four women, each of whom delivered a healthy infant. Our data are in agreement with similar experiences and confirm the activity of GnRH therapy in preventing ovarian failure. PMID- 19129074 TI - A phase II study of 5-fluorouracil/leucovorin in combination with paclitaxel and oxaliplatin as first-line treatment for patients with advanced gastric cancer. AB - The objective of this study was to evaluate the efficacy and safety of the POF regimen (biweekly 5-fluorouracil/leucovorin combined with paclitaxel and oxaliplatin) as first-line treatment for advanced gastric cancer (AGC). Twenty seven previously untreated patients with advanced adenocarcinoma of the gastric or gastroesophageal junction were eligible for this study. The chemotherapy regimen consisted of a 3-hour infusion of paclitaxel (135 mg/m(2)) followed by oxaliplatin (85 mg/m(2)) and leucovorin (400 mg/m(2)), administered simultaneously over a 2-hour infusion period, followed by an infusion of 5 fluorouracil (2400 mg/m(2)) over a 46-hour period. Twenty-one patients had measurable lesions: four complete responses, eight partial responses and seven stable diseases. At a median follow-up of 610 days, median survival was 348 days. Frequent grade 3 to 4 toxicities were: neutropenia (29.6%), stomatitis (7.4%), nausea (7.4%), vomiting (7.4%), hepatic dysfunction (3.7%), and fatigue (18.5%). No treatment-related deaths occurred. The POF regimen appears to be efficacious and is well tolerated in patients with AGC. PMID- 19129075 TI - Contribution of ParE mutation and efflux to ciprofloxacin resistance in Pseudomonas aeruginosa clinical isolates. PMID- 19129076 TI - First report of ciprofloxacin resistance among Klebsiella pneumoniae harbouring the qnrA1 gene and producing SHV-5 extended-spectrum beta-lactamase in Scotland. PMID- 19129077 TI - In vitro activity of amphotericin B against Aspergillus terreus isolates from different countries and regions. PMID- 19129078 TI - Susceptibility to voriconazole, fluconazole, and ketoconazole of yeast isolated from patients with hematological malignancies. PMID- 19129079 TI - Tigecycline for treatment of nosocomial-acquired pneumonia possibly caused by multi-drug resistant strains of Stenotrophomonas maltophilia. PMID- 19129080 TI - Clinical evaluation of affirm VP III in the detection and identification of bacterial vaginosis. PMID- 19129081 TI - Early development of lactic acidosis with short term linezolid treatment in a renal recipient. PMID- 19129082 TI - Association between IL4 (-590), ACE (I)/(D), CCR5 (Delta32), CTLA4 (+49) and IL1 RN (VNTR in intron 2) gene polymorphisms and vitiligo. AB - Vitiligo is a common skin disorder characterized by patterned depigmentation, because of a decrease of melanin pigment resulting from apparent melanocyte loss. The aim of this study was to investigate interleukin 4 (IL4), Angiotensin Converting Enzyme (ACE), C-C Chemocine Receptor 5 (CCR5), Cytotoxic T Lymphocyte associated Antigen Receptor 4 (CTLA4) and Interleukin 1 Receptor Antagonist (IL1 RN) gene polymorphisms in 48 Turkish vitiligo patients and 50 healthy controls. Polymorphisms for the genes ACE insertion(I)/deletion(D), CCR5 (Delta32), IL1-RN (VNTR in intron 2) were detected by PCR methods. IL4 (-590) and CTLA4 (+49) gene polymorphisms were typed using PCR-RFLP methods. No significant differences in either the genotype distribution or allele frequencies of IL4, CCR5 and ACE gene polymorphisms were observed. GG genotype and G allele in CTLA4 genes were found to be significantly higher in vitiligo patients compared to the controls. (0.002, 0.000). CTLA4 (AA) and IL1-RN (1/5) genotypes and 5 allele frequency in the IL1 RN gene were found to be significantly lower in vitiligo patients compared to healthy controls (p: 0.014, 0.015, 0.016, respectively). As a conclusion, CTLA4 and IL1-RN genes might play roles in the genetic etiology of vitiligo. PMID- 19129084 TI - Welcome back to Human Genomics. PMID- 19129083 TI - Fucoidan inhibits UVB-induced MMP-1 promoter expression and down regulation of type I procollagen synthesis in human skin fibroblasts. AB - UVB reduces type I procollagen levels and increases matrix metalloproteinases-1 (MMP-1) levels in human skin and plays a major role in the process of photoaging. We previously reported that fucoidan inhibits UVB-induced MMP-1 expression at the protein and mRNA levels in human skin fibroblasts (HS68). Yet, the effects of fucoidan on UVB-induced MMP-1 promoter activity and type I procollagen have not been investigated. In this study, we assessed the effects of fucoidan on the inhibition of MMP-1 promoter activity and on the increase of type I procollagen synthesis in human skin fibroblasts. Fucoidan treatment significantly inhibited MMP-1 promoter activity compared to UVB irradiation alone. Fucoidan treatment also increased type I procollagen mRNA and protein expression in a dose-dependent manner compared to the control. Our data indicate that fucoidan may prevent UVB induced MMP-1 expression and inhibit down-regulation of type I procollagen synthesis. We suggest that fucoidan may be a potential therapeutic agent to prevent and treat skin photoaging. PMID- 19129085 TI - Guest editorial. Genes and lung cancer. PMID- 19129086 TI - Influence of SLCO1B1 and CYP2C8 gene polymorphisms on rosiglitazone pharmacokinetics in healthy volunteers. AB - Polymorphisms in drug transporter genes and/or drug-metabolising enzyme genes may contribute to inter-individual variability in rosiglitazone pharmacokinetics in humans. We sought to determine the joint effects of polymorphisms in the SLCO1B1 drug transporter gene and the cytochrome P450 ( CYP ) 2C8-metabolising enzyme gene on rosiglitazone pharmacokinetics in healthy volunteers. Healthy Caucasian subjects were prospectively enrolled on the basis of SLCO1B1 521 T > C genotype. Additionally, subjects were genotyped for SLCO1B1 -11187 G > A, -10499 A > C and 388 A > G polymorphisms, and the CYP2C8*3 polymorphism. SLCO1B1 haplotypes and diplotypes were computationally assigned. Rosiglitazone plasma concentrations were determined by high-performance liquid chromatography and analysed using non compartmental methods. The study population consisted of 26 subjects, with a mean age of 33 +/- 9 years, and a mean weight of 66.6 +/- 11.7 kg. There were no significant differences in rosiglitazone pharmacokinetic parameters between SLCO1B1 diplotype groups. Subjects with the CYP2C8*1/*3 genotype ( n = 7), however, had significantly lower rosiglitazone area under the plasma concentration-time curve (AUC) and significantly higher rosiglitazone oral clearance, compared with CYP2C8 wild-type homozygotes ( n = 19). Stepwise linear regression analysis revealed that CYP2C8 genotype ( p = 0.006) and weight ( p = 0.022) were significant predictors of rosiglitazone AUC (overall p = 0.002; R 2 = 41.6 per cent). We concluded that polymorphisms in the CYP2C8 drug-metabolising enzyme gene, but not the SLCO1B1 drug transporter gene, significantly influence rosiglitazone disposition in humans. Future studies examining the influence of CYP2C8 genotypes and haplotypes on thiazolidinedione disposition and response in patients with type 2 diabetes are warranted. PMID- 19129087 TI - A high incidence of polymorphic CYP2C19 variants in archival blood samples from Papua New Guinea. AB - There is considerable inter-ethnic variability in the incidence of CYP2C19 genetic poor metabolisers ( var / var ). About 3 per cent of Caucasians are CYP2C19 var/var . By contrast, an extremely high incidence (70 per cent) is observed in the Melanesian island of Vanuatu. The colonisation of the Pacific Islands is believed to have involved migration through Papua New Guinea (PNG), and hence a high incidence may also be expected in this population. The reported incidence in PNG was only 36 per cent, however. PNG is a country of extensive ethnic diversity, and the incidence of the CYP2C19 var/var in other regional populations of PNG is currently not established. In this study, restriction fragment length polymorphism-polymerase chain reaction analysis of archival blood serum samples was used to determine the prevalence of the CYP2C19*2 and *3 variant alleles in three different ethnic and geographically isolated populations of PNG. In the largest population studied (Iruna), the frequency of both variant CYP2C19 alleles was high (0.37 and 0.34, respectively). Specifically, the frequency of the CYP2C19*3 allele was significantly higher than in the PNG (East Sepik) population reported previously (0.34 vs 0.16; p < 0.0001). In the Iruna population, 48.9 per cent of the samples were homozygous variants for CYP2C19*2 or *3 , which although higher was not statistically different from the East Sepik population (36 per cent). The results of this study indicated that other regional populations of PNG also have a relatively high incidence of the CYP2C19 genetic polymorphism compared with Caucasian populations. The high incidence reported in Vanuatu, however, may be due to genetic drift rather than a PNG founder population, as the Vanuatu population is dominated by the CYP2C19*2 allele, with a lower contribution from the *3 allelic variant. PMID- 19129089 TI - Hardy-Weinberg analysis of a large set of published association studies reveals genotyping error and a deficit of heterozygotes across multiple loci. AB - In genetic association studies, deviation from Hardy-Weinberg equilibrium (HWD) can be due to recent admixture or selection at a locus, but is most commonly due to genotyping errors. In addition to its utility for identifying potential genotyping errors in individual studies, here we report that HWD can be useful in detecting the presence, magnitude and direction of genotyping error across multiple studies. If there is a consistent genotyping error at a given locus, larger studies, in general, will show more evidence for HWD than small studies. As a result, for loci prone to genotyping errors, there will be a correlation between HWD and the study sample size. By contrast, in the absence of consistent genotyping errors, there will be a chance distribution of p- values among studies without correlation with sample size. We calculated the evidence for HWD at 17 separate polymorphic loci investigated in 325 published genetic association studies. In the full set of studies, there was a significant correlation between HWD and locus-standardised sample size ( p = 0.001). For 14/17 of the individual loci, there was a positive correlation between extent of HWD and sample size, with the evidence for two loci ( 5-HTTLPR and CTSD ) rising to the level of statistical significance. Among single nucleotide polymorphisms (SNPs), 15/23 studies that deviated significantly from Hardy-Weinberg equilibrium (HWE) did so because of a deficit of heterozygotes. The inbreeding coefficient (F(is)) is a measure of the degree and direction of deviation from HWE. Among studies investigating SNPs, there was a significant correlation between F(is) and HWD ( R = 0.191; p = 0.002), indicating that the greater the deviation from HWE, the greater the deficit of heterozygotes. By contrast, for repeat variants, only one in five studies that deviated significantly from HWE showed a deficit of heterozygotes and there was no significant correlation between F(is) and HWD. These results indicate the presence of HWD across multiple loci, with the magnitude of the deviation varying substantially from locus to locus. For SNPs, HWD tends to be due to a deficit of heterozygotes, indicating that allelic dropout may be the most prevalent genotyping error. PMID- 19129088 TI - The role of aldehyde dehydrogenase-1 (ALDH1A1) polymorphisms in harmful alcohol consumption in a Finnish population. AB - Liver cystolic aldehyde dehydrogenase 1 (ALDH1A1) has been previously associated with both alcohol dependence and alcohol consumption behaviour, and has been implicated in alcohol-induced flushing and alcohol sensitivity in Caucasians. The present study tested for association between ALDH1A1 and alcohol consumption behaviour and susceptibility to problem drinking or alcohol dependence in Finnish cohorts of unrelated male subjects recruited from alcoholism clinical treatment facilities ( n = 104) and from the general population ( n = 201). All participants completed the Alcohol Use Disorder Identification Test (AUDIT) and were genotyped for eight single nucleotide polymorphisms (SNPs) within or flanking ALDH1A1 . To test for association between alcohol consumption behaviour and these polymorphisms, we used generalised linear models and haplotypic analysis. Three SNPs were nominally associated (rs348449, p = 0.043; rs610529, p = 0.013; rs348479, p = 0.025) with the quantitative AUDIT score, which evaluates alcohol consumption behaviour. Two-locus (rs610529-rs2288087) haplotype analysis increased the strength of association with AUDIT score ( p = 0.0015). Additionally, rs348449 is highly associated with problem drinking (allelic odds ratio [OR] 7.87, 95 per cent confidence interval [CI] 1.67-37.01) but due to the low minor allele frequency (0.01 and 0.07 in controls and problem drinkers, respectively), more samples are required to validate this observation. Conversely, rs348479 ( p = 0.019) and rs610529 (allelic OR 0.65, 95 per cent CI 0.43-0.98; genotypic OR 0.32, 95 per cent CI 0.12-0.84) are implicated in alcohol dependence status. This study provides further evidence for a role for ALDH1A1 in alcohol consumption behaviour, including problem drinking and possibly alcohol dependence, in our Finnish population. PMID- 19129090 TI - Genome-wide identification of genetic determinants for the cytotoxicity of perifosine. AB - Perifosine belongs to the class of alkylphospholipid analogues, which act primarily at the cell membrane, thereby targeting signal transduction pathways. In phase I/II clinical trials, perifosine has induced tumour regression and caused disease stabilisation in a variety of tumour types. The genetic determinants responsible for its cytotoxicity have not been comprehensively studied, however. We performed a genome-wide analysis to identify genes whose expression levels or genotypic variation were correlated with the cytotoxicity of perifosine, using public databases on the US National Cancer Institute (NCI)-60 human cancer cell lines. For demonstrating drug specificity, the NCI Standard Agent Database (including 171 drugs acting through a variety of mechanisms) was used as a control. We identified agents with similar cytotoxicity profiles to that of perifosine in compounds used in the NCI drug screen. Furthermore, Gene Ontology and pathway analyses were carried out on genes more likely to be perifosine specific. The results suggested that genes correlated with perifosine cytotoxicity are connected by certain known pathways that lead to the mitogen activated protein kinase signalling pathway and apoptosis. Biological processes such as 'response to stress', 'inflammatory response' and 'ubiquitin cycle' were enriched among these genes. Three single nucleotide polymorphisms (SNPs) located in CACNA2D1 and EXOC4 were found to be correlated with perifosine cytotoxicity. Our results provided a manageable list of genes whose expression levels or genotypic variation were strongly correlated with the cytotoxcity of perifosine. These genes could be targets for further studies using candidate-gene approaches. The results also provided insights into the pharmacodynamics of perifosine. PMID- 19129091 TI - Mitochondrial DNA sequence variation and neurodegeneration. AB - Mitochondria, the powerhouse of the cell, play a critical role in several metabolic processes and apoptotic pathways. Many lines of evidence suggest that mitochondria have a central role in ageing-related neurodegenerative diseases. Moreover, there is a long history of investigations on mitochondria aimed at identifying genetic markers relating to ageing and neurodegenerative diseases. In this review, some of the major neurodegenerative disorders are highlighted and the role of mitochondrial haplogroups in the pathogenetic cascade leading to these diseases is discussed. PMID- 19129092 TI - A survey of genetic simulation software for population and epidemiological studies. AB - A number of programs have been developed for simulating population genetic and genetic epidemiological data conforming to one of three main algorithmic approaches: 'forwards', 'backwards' and 'sideways'. This review aims to make the reader aware of the range of options currently available to them. While no one program emerges as the best choice in all circumstances, we nominate a set of those which currently appear most promising. PMID- 19129095 TI - Biodiversity research sets sail: showcasing the diversity of marine life. AB - The World Congress on Marine Biodiversity was held in the City of Arts and Sciences, Valencia, from 10 to 15 November 2008, showcasing research on all aspects of marine biodiversity from basic taxonomic exploration to innovative conservation strategies and methods to integrate research into environmental policy. PMID- 19129093 TI - Update on the olfactory receptor (OR) gene superfamily. AB - The olfactory receptor gene (OR) superfamily is the largest in the human genome. The superfamily contains 390 putatively functional genes and 465 pseudogenes arranged into 18 gene families and 300 subfamilies. Even members within the same subfamily are often located on different chromosomes. OR genes are located on all autosomes except chromosome 20, plus the X chromosome but not the Y chromosome. The gene:pseudogene ratio is lowest in human, higher in chimpanzee and highest in rat and mouse--most likely reflecting the greater need of olfaction for survival in the rodent than in the human. The OR genes undergo allelic exclusion, each sensory neurone expressing usually only one odourant receptor allele; the mechanism by which this phenomenon is regulated is not yet understood. The nomenclature system (based on evolutionary divergence of genes into families and subfamilies of the OR gene superfamily) has been designed similarly to that originally used for the CYP gene superfamily. PMID- 19129096 TI - Fatigue alters in vivo function within and between limb muscles during locomotion. AB - Muscle fatigue, a reduction in force as a consequence of exercise, is an important factor for any animal that moves, and can result from both peripheral and/or central mechanisms. Although much is known about whole-limb force generation and activation patterns in fatigued muscles under sustained isometric contractions, little is known about the in vivo dynamics of limb muscle function in relation to whole-body fatigue. Here we show that limb kinematics and contractile function in the lateral (LG) and medial (MG) gastrocnemius of helmeted guineafowl (Numida meleagris) are significantly altered following fatiguing exercise at 2ms-1 on an inclined treadmill. The two most significant findings were that the variation in muscle force generation, measured directly from the muscles' tendons, increased significantly with fatigue, and fascicle shortening in the proximal MG, but not the distal MG, decreased significantly with fatigue. We suggest that the former is a potential mechanism for decreased stability associated with fatigue. The region-specific alteration of fascicle behaviour within the MG as a result of fatigue suggests a complex response to fatigue that probably depends on muscle-aponeurosis and tendon architecture not previously explored. These findings highlight the importance of studying the integrative in vivo dynamics of muscle function in response to fatigue. PMID- 19129097 TI - How much do genetic covariances alter the rate of adaptation? AB - Genetically correlated traits do not evolve independently, and the covariances between traits affect the rate at which a population adapts to a specified selection regime. To measure the impact of genetic covariances on the rate of adaptation, we compare the rate fitness increases given the observed G matrix to the expected rate if all the covariances in the G matrix are set to zero. Using data from the literature, we estimate the effect of genetic covariances in real populations. We find no net tendency for covariances to constrain the rate of adaptation, though the quality and heterogeneity of the data limit the certainty of this result. There are some examples in which covariances strongly constrain the rate of adaptation but these are balanced by counter examples in which covariances facilitate the rate of adaptation; in many cases, covariances have little or no effect. We also discuss how our metric can be used to identify traits or suites of traits whose genetic covariances to other traits have a particularly large impact on the rate of adaptation. PMID- 19129098 TI - Sperm morphology and sperm velocity in passerine birds. AB - Sperm velocity is one of the main determinants of the outcome of sperm competition. Since sperm vary considerably in their morphology between and within species, it seems likely that sperm morphology is associated with sperm velocity. Theory predicts that sperm velocity may be increased by enlarged midpiece (energetic component) or flagellum length (kinetic component), or by particular ratios between sperm components, such as between flagellum length and head size. However, such associations have rarely been found in empirical studies. In a comparative framework in passerine birds, we tested these theoretical predictions both across a wide range of species and within a single family, the New World blackbirds (Icteridae). In both study groups, sperm velocity was influenced by sperm morphology in the predicted direction. Consistent with theoretical models, these results show that selection on sperm morphology and velocity are likely to be concomitant evolutionary forces. PMID- 19129099 TI - When does optional participation allow the evolution of cooperation? AB - Altruistic punishment has been shown to invade when rare if individuals are allowed to opt out of cooperative ventures. Individuals that opt out do not contribute to the common enterprise or derive benefits from it. This result is potentially significant because it offers an explanation for the origin of large scale cooperation in one-shot interactions among unrelated individuals. Here, we show that this result is not a general consequence of optional participation in cooperative activities, but depends on special assumptions about cooperative pay offs. We extend the pay-off structure of optional participation models to consider the effects of economies and diseconomies of scale in public-goods production, rival and non-rival consumption of goods, and different orderings of the pay-offs of freeriding and opting out. This more general model highlights the kinds of pay-offs for which optional participation favours cooperation, and those in which it does not. PMID- 19129100 TI - Mothers produce less aggressive sons with altered immunity when there is a threat of disease during pregnancy. AB - Maternal experience before and during pregnancy is known to play a key role in offspring development. However, the influence of social cues about disease in the maternal environment has not been explored. We indirectly exposed pregnant mice to infected neighbours by housing them next to non-contagious conspecifics infected with Babesia microti. We examined the effect of this indirect immunological exposure on both the females and their adult offspring. Exposed females had higher levels of serum corticosterone and increased kidney growth compared with those with uninfected neighbours. These exposed females subsequently produced offspring that as adults showed an accelerated immune response to B. microti and less aggression in social groups. We suggest that ambient information regarding disease is used adaptively to maximize offspring survival and reproductive success in a challenging environment. Our results shed light on the impact of social information and maternal effects on life histories, and have important consequences for our understanding of epidemiology and individual disease susceptibility in humans and other animals. They also lead us to question the suitability of some laboratory housing conditions during experimental procedures, which may impact negatively upon both animal welfare and the validity of animal science. PMID- 19129101 TI - Divergent trends in anadromous salmonid populations in Norwegian and Scottish rivers. AB - The Atlantic salmon (Salmo salar) is a charismatic anadromous fish of high conservation and economic value. Concerns have been expressed regarding the long term viability of fisheries throughout the species's distributional range because of abundance variations that cannot currently be explained or predicted. Here, we analyse long-term catch data obtained over a wide geographical range and across a range of spatial subscales to understand more fully the factors that drive population abundance. We use rod catch data from 84 Norwegian rivers over 125 years (1876-2000) and 48 Scottish rivers over 51 years (1952-2002). The temporal correlation in catches is very long-term, with trends persisting over several decades. The spatial correlation is relatively short-range, indicating strong local-scale effects on catch. Furthermore, Scottish salmon populations exhibit recent negative trends in contrast to some more positive trends in Norway- especially in the north. PMID- 19129102 TI - Why are (the best) women so good at chess? Participation rates and gender differences in intellectual domains. AB - A popular explanation for the small number of women at the top level of intellectually demanding activities from chess to science appeals to biological differences in the intellectual abilities of men and women. An alternative explanation is that the extreme values in a large sample are likely to be greater than those in a small one. Although the performance of the 100 best German male chess players is better than that of the 100 best German women, we show that 96 per cent of the observed difference would be expected given the much greater number of men who play chess. There is little left for biological or cultural explanations to account for. In science, where there are many more male than female participants, this statistical sampling explanation, rather than differences in intellectual ability, may also be the main reason why women are under-represented at the top end. PMID- 19129103 TI - An exceptionally well-preserved Eocene dolichopodid fly eye: function and evolutionary significance. AB - The exceptionally preserved eyes of an Eocene dolichopodid fly contained in Baltic amber show remarkable detail, including features at micrometre and submicrometre levels. Based on this material, we establish that it is likely that the neural superposition compound eye existed as far back as 45 Ma. The ommatidia have an open rhabdom with a trapezoidal arrangement of seven rhabdomeres. Such a structure is uniquely characteristic of the neural superposition compound eye of present-day flies. Optical analysis reveals that the fossil eyes had a sophisticated and efficient optical system. PMID- 19129104 TI - Liparid and macrourid fishes of the hadal zone: in situ observations of activity and feeding behaviour. AB - Using baited camera landers, the first images of living fishes were recorded in the hadal zone (6000-11000 m) in the Pacific Ocean. The widespread abyssal macrourid Coryphaenoides yaquinae was observed at a new depth record of approximately 7000 m in the Japan Trench. Two endemic species of liparid were observed at similar depths: Pseudoliparis amblystomopsis in the Japan Trench and Notoliparis kermadecensis in the Kermadec Trench. From these observations, we have documented swimming and feeding behaviour of these species and derived the first estimates of hadal fish abundance. The liparids intercepted bait within 100 200 min but were observed to preferentially feed on scavenging amphipods. Notoliparis kermadecensis act as top predators in the hadal food web, exhibiting up to nine suction-feeding events per minute. Both species showed distinctive swimming gaits: P. amblystomopsis (mean length 22.5 cm) displayed a mean tail beat frequency of 0.47 Hz and mean caudal:pectoral frequency ratio of 0.76, whereas N. kermadecensis (mean length 31.5 cm) displayed respective values of 1.04 and 2.08 Hz. Despite living at extreme depths, these endemic liparids exhibit similar activity levels compared with shallow-water liparids. PMID- 19129105 TI - Parasite manipulation of brain monoamines in California killifish (Fundulus parvipinnis) by the trematode Euhaplorchis californiensis. AB - California killifish (Fundulus parvipinnis) infected with the brain-encysting trematode Euhaplorchis californiensis display conspicuous swimming behaviours rendering them more susceptible to predation by avian final hosts. Heavily infected killifish grow and reproduce normally, despite having thousands of cysts inside their braincases. This suggests that E. californiensis affects only specific locomotory behaviours. We hypothesised that changes in the serotonin and dopamine metabolism, essential for controlling locomotion and arousal may underlie this behaviour modification. We employed micropunch dissection and HPLC to analyse monoamine and monoamine metabolite concentrations in the brain regions of uninfected and experimentally infected fish. The parasites exerted density dependent changes in monoaminergic activity distinct from those exhibited by fish subjected to stress. Specifically, E. californiensis inhibited a normally occurring, stress-induced elevation of serotonergic metabolism in the raphae nuclei. This effect was particularly evident in the experimentally infected fish, whose low-density infections were concentrated on the brainstem. Furthermore, high E. californiensis density was associated with increased dopaminergic activity in the hypothalamus and decreased serotonergic activity in the hippocampus. In conclusion, the altered monoaminergic metabolism may explain behavioural differences leading to increased predation of the infected killifish by their final host predators. PMID- 19129106 TI - Environmental heterogeneity, genotype-by-environment interactions and the reliability of sexual traits as indicators of mate quality. AB - Exaggerated sexual displays are often supposed to indicate the indirect benefits females may receive from sexual reproduction with displaying males, but empirical evidence for positive relationships between the genetic quality and sexual trait quality is scant. The explanation for this might lie in the fact that mixing of reproductive individuals whose development has been influenced by genotype-by environment interactions (GEIs) can blur the relationship between the individual male genetic quality and phenotype as perceived by females. Strong GEIs can generate an ecological crossover, where different genotypes are superior in environments that are separated either in space or time. Here, we use a stochastic simulation model to show that even a weak GEI, which does not generate an obvious ecological crossover, can neutralize or even reverse the relationship between genetic quality and sexual trait size in the presence of environmental heterogeneity during development. Our model highlights the importance of developmental selection in evolution of traits and allows us to predict the situations in which sexual displays might not be reliable indicators of genetic quality. PMID- 19129108 TI - Food-density-dependent inefficiency in animals with a gut as a stabilizing mechanism in trophic dynamics. AB - Animals with a gut, when confronted with food supplied ad libitum, can elevate their ingestion rates and inefficiently use the material they ingest. As a consequence, assimilation efficiency (AE) declines, resulting in food-density dependent inefficiency (f-DDI). A model describing these processes shows that f DDI can dampen the consequences of oscillations in food abundance that may occur in response to external stochastic (e.g. climatic) forcing both with respect to production and timing. This response is illustrated with a simple planktonic food chain of a phytoplankter and two consumers. The assumption of a fixed gut transit time, consistent with the traditional model descriptions of a fixed AE, produces predator-prey oscillations. By contrast, simulations using a model showing f-DDI (behaving in accordance with the experimental data) cushion not only the impact of such oscillations but also the effects of the removal of intermediate grazers in the food chain. The operation of f-DDI affects other trophic interactions through changes in the nutrient regeneration and the voiding rates. The extent to which f-DDI operates in nature needs valuation, followed by the appropriate construction of consumer-based ecosystem models. PMID- 19129109 TI - A bizarre Cretaceous theropod dinosaur from Patagonia and the evolution of Gondwanan dromaeosaurids. AB - Fossils of a predatory dinosaur provide novel information about the evolution of unenlagiines, a poorly known group of dromaeosaurid theropods from Gondwana. The new dinosaur is the largest dromaeosaurid yet discovered in the Southern Hemisphere and depicts bizarre cranial and postcranial features. Its long and low snout bears numerous, small-sized conical teeth, a condition resembling spinosaurid theropods. Its short forearms depart from the characteristically long armed condition of all dromaeosaurids and their close avian relatives. The new discovery amplifies the range of morphological disparity among unenlagiines, demonstrating that by the end of the Cretaceous this clade included large, short armed forms alongside crow-sized, long-armed, possibly flying representatives. The new dinosaur is the youngest record of dromaeosaurids from Gondwana and represents a previously unrecognized lineage of large predators in Late Cretaceous dinosaur faunas mainly dominated by abelisaurid theropods. PMID- 19129110 TI - Extreme convergence in stick insect evolution: phylogenetic placement of the Lord Howe Island tree lobster. AB - The 'tree lobsters' are an enigmatic group of robust, ground-dwelling stick insects (order Phasmatodea) from the subfamily Eurycanthinae, distributed in New Guinea, New Caledonia and associated islands. Its most famous member is the Lord Howe Island stick insect Dryococelus australis (Montrouzier), which was believed to have become extinct but was rediscovered in 2001 and is considered to be one of the rarest insects in the world. To resolve the evolutionary position of Dryococelus, we constructed a phylogeny from approximately 2.4 kb of mitochondrial and nuclear sequence data from representatives of all major phasmatodean lineages. Our data placed Dryococelus and the New Caledonian tree lobsters outside the New Guinean Eurycanthinae as members of an unrelated Australasian stick insect clade, the Lanceocercata. These results suggest a convergent origin of the 'tree lobster' body form. Our reanalysis of tree lobster characters provides additional support for our hypothesis of convergent evolution. We conclude that the phenotypic traits leading to the traditional classification are convergent adaptations to ground-living behaviour. Our molecular dating analyses indicate an ancient divergence (more than 22 Myr ago) between Dryococelus and its Australian relatives. Hence, Dryococelus represents a long-standing separate evolutionary lineage within the stick insects and must be regarded as a key taxon to protect with respect to phasmatodean diversity. PMID- 19129111 TI - Earthworm genomes, genes and proteins: the (re)discovery of Darwin's worms. AB - Small incremental biological change, winnowed by natural selection over geological time scales to produce large consequences, was Darwin's singular insight that revolutionized the life sciences. His publications after 1859, including the 'earthworm book', were all written to amplify and support the evolutionary theory presented in the Origin. Darwin was unable to provide a physical basis for the inheritance of favoured traits because of the absence of genetic knowledge that much later led to the 'modern synthesis'. Mistaken though he was in advocating systemic 'gemmules' as agents of inheritance, Darwin was perceptive in seeking to underpin his core vision with concrete factors that both determine the nature of a trait in one generation and convey it to subsequent generations. This brief review evaluates the molecular genetic literature on earthworms published during the last decade, and casts light on the specific aspects of earthworm evolutionary biology that more or less engaged Darwin: (i) biogeography, (ii) species diversity, (iii) local adaptations and (iv) sensitivity. We predict that the current understanding will deepen with the announcement of a draft earthworm genome in Darwin's bicentenary year, 2009. Subsequently, the earthworm may be elevated from the status of a soil sentinel to that elusive entity, an ecologically relevant genetic model organism. PMID- 19129112 TI - Problems detecting density-dependent diversification on phylogenies. PMID- 19129113 TI - The relationship between social behaviour and habitat familiarity in African elephants (Loxodonta africana). AB - Social associations with conspecifics can expedite animals' acclimation to novel environments. However, the benefits gained from sociality may change as the habitat becomes familiar. Furthermore, the particular individuals with whom animals associate upon arrival at a new place, familiar conspecifics or knowledgeable unfamiliar residents, may influence the type of information they acquire about their new home. To examine animals' social dynamics in novel habitats, we studied the social behaviour of African elephants (Loxodonta africana) translocated into a novel environment. We found that the translocated elephants' association with conspecifics decreased over time supporting our hypothesis that sociality provides added benefits in novel environments. In addition, we found a positive correlation between body condition and social association, suggesting that elephants gain direct benefits from sociality. Furthermore, the translocated elephants associated significantly less than expected with the local residents and more than expected with familiar, but not necessarily genetically related, translocated elephants. The social segregation between the translocated and resident elephants declined over time, suggesting that elephants can integrate into an existing social setting. Knowledge of the relationship between sociality and habitat familiarity is highly important in our constantly changing world to both conservation practice and our understanding of animals' behaviour in novel environments. PMID- 19129114 TI - Major histocompatibility complex (MHC) heterozygote superiority to natural multi parasite infections in the water vole (Arvicola terrestris). AB - The fundamental role of the major histocompatibility complex (MHC) in immune recognition has led to a general consensus that the characteristically high levels of functional polymorphism at MHC genes is maintained by balancing selection operating through host-parasite coevolution. However, the actual mechanism by which selection operates is unclear. Two hypotheses have been proposed: overdominance (or heterozygote superiority) and negative frequency dependent selection. Evidence for these hypotheses was evaluated by examining MHC parasite relationships in an island population of water voles (Arvicola terrestris). Generalized linear mixed models were used to examine whether individual variation at an MHC class II DRB locus explained variation in the individual burdens of five different parasites. MHC genotype explained a significant amount of variation in the burden of gamasid mites, fleas (Megabothris walkeri) and nymphs of sheep ticks (Ixodes ricinus). Additionally, MHC heterozygotes were simultaneously co-infected by fewer parasite types than homozygotes. In each case where an MHC-dependent effect on parasite burden was resolved, the heterozygote genotype was associated with fewer parasites, and the heterozygote outperformed each homozygote in two of three cases, suggesting an overall superiority against parasitism for MHC heterozygote genotypes. This is the first demonstration of MHC heterozygote superiority against multiple parasites in a natural population, a mechanism that could help maintain high levels of functional MHC genetic diversity in natural populations. PMID- 19129116 TI - Survival costs of reproduction vary with age in North American red squirrels. AB - The costs of reproduction are expected to be higher under unfavourable conditions, so that breeding in years of low food supply should have important costs. In addition, the costs of reproduction may be contingent on the age of individuals, and young growing and old senescent individuals should suffer higher costs than the prime-age ones. We tested these predictions by investigating the costs of reproduction as a function of food availability and age in female North American red squirrels using the long-term data on survival and reproduction. We found that the costs of reproduction were independent of food supply, and we did not detect any trade-off between the current and future reproduction. We also did not detect any survival cost of reproduction for the prime-age females, but found evidence for survival costs in yearlings and old (6 years or above) females with successfully breeding individuals having a lower chance of survival compared with unsuccessful or non-breeding ones. These results supported our prediction that the costs of reproduction depended on the age of female red squirrels and were higher in young growing and old senescent individuals. Our study also indicated that, in contrast to large herbivores, heterogeneity in individual quality and viability selection in red squirrels do not affect the study of trade-offs and of the age variation in life-history traits. PMID- 19129115 TI - Sex-specific variation in the emphasis, inducibility and timing of the post mating immune response in Drosophila melanogaster. AB - Ecological immunology attempts to explain variation in immune function. Much of this work makes predictions about how potential hosts should invest in overall immunity. However, this 'overall' perspective under-emphasizes other critical aspects, such as the specificity, inducibility and timing of an immune response. Here, we investigate these aspects by examining gene regulation across several immune system components in both male and female Drosophila melanogaster prior to and after mating. To elucidate potentially important temporal dynamics, we also assayed several genes over time. We found that males and females emphasized different components of their immune system, however overall investment was similar. Specifically, the sexes emphasized different gene paralogues within major gene families, and males tended to invest more in gram-negative defence. By contrast, the inducibility of the immune response was both transient (lasting approx. 24 hours) and equal between the sexes. Furthermore, mating tended to induce humoral gene upregulation, while cell-mediated genes were unaffected. Within the humoral system, gram-negative bacterial defence genes exhibited a greater inducibility than those associated with fungal or gram-positive bacterial defence. Our results suggest that variation in the effectiveness of the immune response between the sexes may be driven by differences in emphasis rather than overall investment. PMID- 19129117 TI - Oxygen hypothesis of polar gigantism not supported by performance of Antarctic pycnogonids in hypoxia. AB - Compared to temperate and tropical relatives, some high-latitude marine species are large-bodied, a phenomenon known as polar gigantism. A leading hypothesis on the physiological basis of gigantism posits that, in polar water, high oxygen availability coupled to low metabolic rates relieves constraints on oxygen transport and allows the evolution of large body size. Here, we test the oxygen hypothesis using Antarctic pycnogonids, which have been evolving in very cold conditions (-1.8-0 degrees C) for several million years and contain spectacular examples of gigantism. Pycnogonids from 12 species, spanning three orders of magnitude in body mass, were collected from McMurdo Sound, Antarctica. Individual sea spiders were forced into activity and their performance was measured at different experimental levels of dissolved oxygen (DO). The oxygen hypothesis predicts that, all else being equal, large pycnogonids should perform disproportionately poorly in hypoxia, an outcome that would appear as a statistically significant interaction between body size and oxygen level. In fact, although we found large effects of DO on performance, and substantial interspecific variability in oxygen sensitivity, there was no evidence for sizexDO interactions. These data do not support the oxygen hypothesis of Antarctic pycnogonid gigantism and suggest that explanations must be sought in other ecological or evolutionary processes. PMID- 19129118 TI - Cnidarian internal stinging mechanism. AB - Stinging mechanisms generally deliver venomous compounds to external targets. However, nematocysts, the microscopic stinging organelles that are common to all members of the phylum Cnidaria, occur and act in both external and internal tissue structures. This is the first report of such an internal piercing mechanism. This mechanism identifies prey items within the body cavity of the sea anemone and actively injects them with cytolytic venom compounds. Internal tissues isolated from sea anemones caused the degradation of live Artemia salina nauplii in vitro. When examined, the nauplii were found to be pierced by discharged nematocysts. This phenomenon is suggested to aid digestive phagocytic processes in a predator otherwise lacking the means to masticate its prey. PMID- 19129121 TI - Oscillations in continuous culture populations of Streptococcus pneumoniae: population dynamics and the evolution of clonal suicide. AB - Agents that kill or induce suicide in the organisms that produce them or other individuals of the same genotype are intriguing puzzles for ecologists and evolutionary biologists. When those organisms are pathogenic bacteria, these suicidal toxins have the added appeal as candidates for the development of narrow spectrum antibiotics to kill the pathogens that produce them.We show that when clinical as well as laboratory strains of Streptococcus pneumoniae are maintained in continuous culture (chemostats), their densities oscillate by as much as five orders of magnitude with an apparently constant period. This dynamic, which is unanticipated for single clones of bacteria in chemostats, can be attributed to population-wide die-offs and recoveries. Using a combination of mathematical models and experiments with S. pneumoniae, we present evidence that these die offs can be attributed to the autocatalytic production of a toxin that lyses or induces autolysis in members of the clone that produces it. This toxin, which our evidence indicates is a protein, appears to be novel; S. pneumoniae genetic constructs knocked out for lytA and other genes coding for known candidates for this agent oscillate in chemostat culture. Since this toxin lyses different strains of S. pneumoniae as well as other closely related species of Streptococcus, we propose that its ecological role is as an allelopathic agent. Using a mathematical model, we explore the conditions under which toxins that kill members of the same clone that produces them can prevent established populations from invasion by different strains of the same or other species. We postulate that the production of the toxin observed here as well as other bacteria-produced toxins that kill members of the same genotype, 'clonal suicide', evolved and are maintained to prevent colonization of established populations by different strains of the same and closely related species. PMID- 19129122 TI - Honest sexual signalling mediated by parasite and testosterone effects on oxidative balance. AB - Extravagant ornaments evolved to advertise their bearers' quality, the honesty of the signal being ensured by the cost paid to produce or maintain it. The oxidation handicap hypothesis (OHH) proposes that a main cost of testosterone dependent ornamentation is oxidative stress, a condition whereby the production of reactive oxygen and nitrogen species (ROS/RNS) overwhelms the capacity of antioxidant defences. ROS/RNS are unstable, very reactive by-products of normal metabolic processes that can cause extensive damage to key biomolecules (cellular proteins, lipids and DNA). Oxidative stress has been implicated in the aetiology of many diseases and could link ornamentation and genetic variation in fitness related traits. We tested the OHH in a free-living bird, the red grouse. We show that elevated testosterone enhanced ornamentation and increased circulating antioxidant levels, but caused oxidative damage. Males with smaller ornaments suffered more oxidative damage than those with larger ornaments when forced to increase testosterone levels, consistent with a handicap mechanism. Parasites depleted antioxidant defences, caused oxidative damage and reduced ornament expression. Oxidative damage extent and the ability of males to increase antioxidant defences also explained the impacts of testosterone and parasites on ornamentation within treatment groups. Because oxidative stress is intimately linked to immune function, parasite resistance and fitness, it provides a reliable currency in the trade-off between individual health and ornamentation. The costs induced by oxidative stress can apply to a wide range of signals, which are testosterone-dependent or coloured by pigments with antioxidant properties. PMID- 19129123 TI - Distance from Africa, not climate, explains within-population phenotypic diversity in humans. AB - The relative importance of ancient demography and climate in determining worldwide patterns of human within-population phenotypic diversity is still open to debate. Several morphometric traits have been argued to be under selection by climatic factors, but it is unclear whether climate affects the global decline in morphological diversity with increasing geographical distance from sub-Saharan Africa. Using a large database of male and female skull measurements, we apply an explicit framework to quantify the relative role of climate and distance from Africa. We show that distance from sub-Saharan Africa is the sole determinant of human within-population phenotypic diversity, while climate plays no role. By selecting the most informative set of traits, it was possible to explain over half of the worldwide variation in phenotypic diversity. These results mirror those previously obtained for genetic markers and show that 'bones and molecules' are in perfect agreement for humans. PMID- 19129124 TI - Cryptic preference for MHC-dissimilar females in male red junglefowl, Gallus gallus. AB - An increasing number of studies test the idea that females increase offspring fitness by biasing fertilization in favour of genetically compatible partners; however, few have investigated or controlled for corresponding preferences in males. Here, we experimentally test whether male red junglefowl, Gallus gallus, prefer genetically compatible females, measured by similarity at the major histocompatibility complex (MHC), a key gene complex in vertebrate immune function. Theory predicts that because some degree of MHC heterozygosity favours viability, individuals should prefer partners that carry MHC alleles different from their own. While male fowl showed no preference when simultaneously presented with an MHC-similar and an MHC-dissimilar female, they showed a 'cryptic' preference, by allocating more sperm to the most MHC-dissimilar of two sequentially presented females. These results provide the first experimental evidence that males might respond to the MHC similarity of a female through differential ejaculate expenditure. By revealing that cryptic male behaviours may bias fertilization success in favour of genetically compatible partners, this study demonstrates the need to experimentally disentangle male and female effects when studying preferences for genetically compatible partners. PMID- 19129125 TI - Voice pitch alters mate-choice-relevant perception in hunter-gatherers. AB - In humans, voice pitch is thought to be a cue of underlying quality and an important criterion for mate choice, but data from non-Western cultures have not been provided. Here we test attributions to and preferences for voices with raised and lowered pitch in hunter-gatherers. Using a forced-choice playback experiment, we found that both men and women viewed lower pitched voices in the opposite sex as being better at acquiring resources (e.g. hunting and gathering). While men preferred higher pitched women's voices as marriage partners, women showed no overall preference for voice pitch in men. However, women who were currently breastfeeding had stronger preferences for higher pitched male voices whereas women not currently breastfeeding preferred lower pitched voices. As testosterone is considered a costly signal associated with dominance, heritable immunity to infection and low paternal investment, women's preferences potentially reflect a trade-off between securing good genes and paternal investment. Men's preferences for higher pitched female voices are probably due to an evolved preference for markers of fecundity, reflected in voice pitch. PMID- 19129126 TI - Echolocating bats emit a highly directional sonar sound beam in the field. AB - Bats use echolocation or biosonar to navigate and find prey at night. They emit short ultrasonic calls and listen for reflected echoes. The beam width of the calls is central to the function of the sonar, but directionality of echolocation calls has never been measured from bats flying in the wild. We used a microphone array to record sounds and determine horizontal directionality for echolocation calls of the trawling Daubenton's bat, Myotis daubentonii, flying over a pond in its natural habitat. Myotis daubentonii emitted highly directional calls in the field. Directionality increased with frequency. At 40kHz half-amplitude angle was 25 degrees , decreasing to 14 degrees at 75kHz. In the laboratory, M. daubentonii emitted less intense and less directional calls. At 55kHz half-amplitude angle was 40 degrees in the laboratory versus 20 degrees in the field. The relationship between frequency and directionality can be explained by the simple piston model. The model also suggests that the increase in the emitted intensity in the field is caused by the increased directionality, focusing sound energy in the forward direction. The bat may increase directionality by opening the mouth wider to emit a louder, narrower beam in the wild. PMID- 19129127 TI - Effects of influenza A virus infection on migrating mallard ducks. AB - The natural reservoir of influenza A virus is waterfowl, particularly dabbling ducks (genus Anas). Although it has long been assumed that waterfowl are asymptomatic carriers of the virus, a recent study found that low-pathogenic avian influenza (LPAI) infection in Bewick's swans (Cygnus columbianus bewickii) negatively affected stopover time, body mass and feeding behaviour. In the present study, we investigated whether LPAI infection incurred ecological or physiological costs to migratory mallards (Anas platyrhynchos) in terms of body mass loss and staging time, and whether such costs could influence the likelihood for long-distance dispersal of the avian influenza virus by individual ducks. During the autumn migrations of 2002-2007, we collected faecal samples (n=10918) and biometric data from mallards captured and banded at Ottenby, a major staging site in a flyway connecting breeding and wintering areas of European waterfowl. Body mass was significantly lower in infected ducks than in uninfected ducks (mean difference almost 20 g over all groups), and the amount of virus shed by infected juveniles was negatively correlated with body mass. There was no general effect of infection on staging time, except for juveniles in September, in which birds that shed fewer viruses stayed shorter than birds that shed more viruses. LPAI infection did not affect speed or distance of subsequent migration. The data from recaptured individuals showed that the maximum duration of infection was on average 8.3 days (s.e. 0.5), with a mean minimum duration of virus shedding of only 3.1 days (s.e. 0.1). Shedding time decreased during the season, suggesting that mallards acquire transient immunity for LPAI infection. In conclusion, deteriorated body mass following infection was detected, but it remains to be seen whether this has more long-term fitness effects. The short virus shedding time suggests that individual mallards are less likely to spread the virus at continental or intercontinental scales. PMID- 19129128 TI - Symbiotic bacteria enable insect to use a nutritionally inadequate diet. AB - Animals generally require a dietary supply of various nutrients (vitamins, essential amino acids, etc.) because their biosynthetic capabilities are limited. The capacity of aphids to use plant phloem sap, with low essential amino acid content, has been attributed to their symbiotic bacteria, Buchnera aphidicola, which can synthesize these nutrients; but this has not been demonstrated empirically. We demonstrate here that phloem sap obtained from the severed stylets of pea aphids Acyrthosiphon pisum feeding on Vicia faba plants generally provided inadequate amounts of at least one essential amino acid to support aphid growth. Complementary analyses using aphids reared on chemically defined diets with each amino acid individually omitted revealed that the capacity of the symbiotic bacterium B. aphidicola to synthesize essential amino acids exceeded the dietary deficit of all phloem amino acids except methionine. It is proposed that this shortfall of methionine was met by aphid usage of the non-protein amino acid 5-methylmethionine in the phloem sap. This study provides the first quantitative demonstration that bacterial symbiosis can meet the nutritional demand of plant-reared aphids. It shows how symbiosis with micro-organisms has enabled this group of animals to escape from the constraint of requiring a balanced dietary supply of amino acids. PMID- 19129129 TI - A female songbird out-sings male conspecifics during simulated territorial intrusions. AB - While birdsong is a model system for animal communication studies, our knowledge is derived primarily from the study of only one sex and is therefore incomplete. The study of song in a role-reversed species would provide a unique opportunity to study selective pressures and mechanisms specific to females, and to test the robustness of current theories in an empirically novel manner. We investigated function of female song in stripe-headed sparrows (Aimophila r. ruficauda), a Neotropical, duetting passerine, and found that during simulated territorial intrusions by a female, male or duetting pair, females: (i) sang more than males to same-sex and duet playback, (ii) played a leading singing role in all contexts, and (iii) showed a longer term song response than males. These results suggest that females sing competitively against other females, and that intrasexual selection may be greater among females than among males. This is the first songbird study to show a stronger vocal role in territory defence for females than males. Stripe-headed sparrows are group-living cooperative breeders, and preliminary data suggest that polyandry and/or resource defence may explain strong female singing behaviour. Stripe-headed sparrows may be a useful study species for expanding our knowledge of vocal communication in female animals. PMID- 19129130 TI - Diversification trajectories and evolutionary life-history traits in early sharks and batoids. AB - Different interpretations on the timing of early diversification and radiation of modern sharks and batoids (Neoselachii) in the Earth's history exist and are related to discrepancies in taxonomic and phylogenetic interpretations favouring a Late Triassic or earliest Jurassic diversification and subsequent radiation event, respectively. Sampling standardization based on pooled taxonomic occurrences made it possible to overcome the problem of a much richer neoselachian record in the Late Jurassic than earlier on. The standardized pattern of genus richness is one of low and fairly constant diversity in the Late Triassic and earliest Jurassic with a steep rise in the Toarcian (ca 180Myr ago), representing the maximum diversification rate in the Jurassic towards a Middle and Late Jurassic plateau. The major Toarcian diversification agrees with the conclusions based on phylogenetic analyses, but is in conflict with older interpretations based on raw data. Early Jurassic expansion of neoselachians was opportunistic in the aftermath of the end-Triassic mass extinction and the reasons for their rapid diversification and radiation probably include small body size, short lifespans and oviparity, enabling faster ecological reorganizations and innovations in body plans for adapting to changing environmental conditions. PMID- 19129131 TI - Pollinator experience, neophobia and the evolution of flowering time. AB - Environmental changes, such as current climate warming, can exert directional selection on reproductive phenology. In plants, evolution of earlier flowering requires that the individuals bearing genes for early flowering successfully reproduce; for non-selfing, zoophilous species, this means that early flowering individuals must be visited by pollinators. In a laboratory experiment with artificial flowers, we presented captive bumble-bees (Bombus impatiens) with flower arrays representing stages in the phenological progression of a two species plant community: Bees that had been foraging on flowers of one colour were confronted with increasing numbers of flowers of a second colour. Early flowering individuals of the second 'species' were significantly under-visited, because bees avoided unfamiliar flowers, particularly when these were rare. We incorporated these aspects of bee foraging behaviour (neophobia and positive frequency dependence) in a simulation model of flowering-time evolution for a plant population experiencing selection against late flowering. Unlike simple frequency dependence, a lag in pollinator visitation prevented the plant population from responding to selection and led to declines in population size. Pollinator behaviour thus has the potential to constrain evolutionary adjustments of flowering phenology. PMID- 19129132 TI - What does testosterone do for red deer males? AB - Testosterone has been proposed to have a dual effect, enhancing sexual traits while depressing parasite resistance in males. Here, we test this hypothesis in red deer, examining males from captive populations during the whole annual cycle and males from natural populations during the breeding season. We first explored the effects of body size, age and sampling date on testosterone to avoid confounding effects. Our results show that in captive populations seasonal changes in testosterone levels were mirrored by changes in testes size, and that during the rut there was a strong correlation between both. In natural populations, males with higher testosterone levels had larger testes, improved sperm quality, smaller burr diameter, stronger antlers, higher haematocrit levels, and increased nematode parasite load. By contrast, no significant relationship was found between testosterone and spleen size or tick parasite load. We conclude that testosterone (i) improves males' reproductive investment and physical stamina, (ii) improves antler strength but reduces burr diameter, and (iii) imposes a cost in terms of depressed parasite resistance. PMID- 19129133 TI - Wholes and subparts in visual processing of human agency. AB - The human visual system is remarkably sensitive to stimuli conveying actions, for example the fighting action between two agents. A central unresolved question is whether each agent is processed as a whole in one stage, or as subparts (e.g. limbs) that are assembled into an agent at a later stage. We measured the perceptual impact of perturbing an agent either by scrambling individual limbs while leaving the relationship between limbs unaffected or conversely by scrambling the relationship between limbs while leaving individual limbs unaffected. Our measurements differed for the two conditions, providing conclusive evidence against a one-stage model. The results were instead consistent with a two-stage processing pathway: an early bottom-up stage where local motion signals are integrated to reconstruct individual limbs (arms and legs), and a subsequent top-down stage where limbs are combined to represent whole agents. PMID- 19129134 TI - The optimal coyness game. AB - In many animal species, females will benefit if they can secure their mate's help in raising their young. It has been suggested that they can achieve this by being coy (i.e. reluctant to mate) when courted, because this gives them time to assess a prospective mate's helpfulness and hence allows them to reject non-helpful males. According to this view, coyness should (i) reflect a trade-off between information gain and time lost on the part of the female, and (ii) be subject to an evolutionary feedback between optimal female coyness and male helping behaviour. Previous theory has considered each of these aspects in isolation. By contrast, here we present a comprehensive game theory model of this situation, leading to qualitatively new insights. We predict that a high degree of coyness should be associated with a high encounter rate during mate search, with an intermediate rate of information gain during mate inspection and with an intermediate dependence of reproduction on male help. Strongly biased sex ratios, however, preclude coyness. Due to the mutual feedback between coyness and helpfulness in our model, alternatively stable evolutionary outcomes (with or without coyness) are possible under broad conditions. We also discuss alternative interpretations of coyness. PMID- 19129135 TI - No energetic cost of anthropogenic disturbance in a songbird. AB - Anthropogenic or natural disturbances can have a significant impact on wild animals. Therefore, understanding when, how and what type of human and natural events disturb animals is a central problem in wildlife conservation. However, it can be difficult to identify which particular environmental stressor affects an individual most. We use heart rate telemetry to quantify the energy expenditure associated with different types of human-mediated and natural disturbances in a breeding passerine, the white-eyed vireo (Vireo griseus). We fitted 0.5g heart rate transmitters to 14 male vireos and continuously recorded heart rate and activity for two days and three nights on a military installation. We calibrated heart rate to energy expenditure for five additional males using an open-flow, push-through respirometry system showing that heart rate predicted 74 per cent of energy expenditure. We conducted standardized disturbance trials in the field to experimentally simulate a natural stressor (predator presence) and two anthropogenic stressors. Although birds initially showed behavioural and heart rate reactions to some disturbances, we could not detect an overall increase in energy expenditure during 1- or 4-hours disturbances. Similarly, overall activity rates were unaltered between control and experimental periods, and birds continued to perform parental duties despite the experimental disturbances. We suggest that vireos quickly determined that disturbances were non-threatening and thus showed no (costly) physiological response. We hypothesize that the lack of a significant response to disturbance in vireos is adaptive and may be representative of animals with fast life histories (e.g. short lifespan, high reproductive output) so as to maximize energy allocation to reproduction. Conversely, we predict that energetic cost of human-mediated disturbances will be significant in slow-living animals. PMID- 19129136 TI - Contact heterogeneity in deer mice: implications for Sin Nombre virus transmission. AB - Heterogeneities within disease hosts suggest that not all individuals have the same probability of transmitting disease or becoming infected. This heterogeneity is thought to be due to dissimilarity in susceptibility and exposure among hosts. As such, it has been proposed that many host-pathogen systems follow the general pattern whereby a small fraction of the population accounts for a large fraction of the pathogen transmission. This disparity in transmission dynamics is often referred to as '20/80 Rule', i.e. approximately 20 per cent of the hosts are responsible for 80 per cent of pathogen transmission. We investigated the role of heterogeneity in contact rates among potential hosts of a directly transmitted pathogen by examining Sin Nombre virus (SNV) in deer mice (Peromyscus maniculatus). Using foraging arenas and powder marking, we documented contacts between wild deer mice in Great Basin Desert, central Utah. Our findings demonstrated heterogeneity among deer mice, both in frequency and in duration of contacts with other deer mice. Contact dynamics appear to follow the general pattern that a minority of the population accounts for a majority of the contacts. We found that 20 per cent of individuals in the population were responsible for roughly 80 per cent of the contacts observed. Larger-bodied individuals appear to be the functional group with the greatest SNV transmission potential. Contrary to our predictions, transmission potential was not influenced by breeding condition or sex. PMID- 19129137 TI - Reproductive competition in the bumble-bee Bombus terrestris: do workers advertise sterility? AB - Reproductive competition in social insects is generally mediated through specific fertility pheromones. By analysing Dufour's gland secretion in queens and workers of Bombus terrestris under varying social conditions, we demonstrate here that the volatile constituents of the secretion exhibit a context-dependent composition. The secretion of egg-laying queens is composed of a series of aliphatic hydrocarbons (alkanes and alkenes), while that of sterile workers contains in addition octyl esters, dominated by octyl hexadecanoate and octyl oleate. These esters disappear in workers with developed ovaries, whether queenright (QR) or queenless (QL), rendering their secretion queen-like. This constitutes an unusual case in which the sterile caste, rather than the fertile one, possesses extra components. Individually isolated (socially deprived) workers developed ovaries successfully, but failed to oviposit, and still possessed the octyl esters. Thus, whereas social interactions are not needed in order to develop ovaries, they appear essential for oviposition and compositional changes in Dufour's gland secretion (ester disappearance). The apparent link between high ester levels and an inability to lay eggs lends credence to the hypothesis that these esters signal functional sterility. We hypothesize that by producing a sterility-specific secretion, workers signal that 'I am out of the competition', and therefore are not attacked, either by the queen or by the reproductive workers. This enables proper colony function and brood care, in particular sexual brood, even under the chaotic conditions of the competition phase. PMID- 19129138 TI - Manipulating grooming by decreasing ectoparasite load causes unpredicted changes in antagonism. AB - It is thought that allogrooming is practised strategically in order to establish, maintain and reinforce social bonds between group members, exchanging one altruistic behaviour for a different form of reciprocated benefit at a later date. Correlational evidence supports this, but evidence of causality is lacking. We reduced parasite loads in eight meerkat Suricata suricatta groups, generating a substantial decrease in grooming. Contrary to the predictions, overall antagonism did not increase. However, within group networks, grooming increased towards individuals who increased their antagonism. This was restricted to antagonism focused on social position, rather than access to physical resources. The treatment also increased an alternative placatory behaviour: unprompted submissions. Following treatment, individuals performed higher rates of guarding and marking behaviours, suggesting that they were stressed. A reduction in opportunity to mediate stress through grooming may explain local rises in antagonism and corresponding increases in placatory behaviours. We suggest that meerkats use allogrooming (and submissions) as a facultative response to antagonism, rather than a pre-emptive strategy to avert it by establishing a network of associations, as has been suggested for primates. PMID- 19129139 TI - Calling in the gap: competition or cooperation in littermates' begging behaviour? AB - Offspring are frequently raised alongside their siblings and are provisioned early in life by adults. Adult provisioning is stimulated by offspring begging, but it is unclear how each offspring should beg, given the begging behaviour of their siblings. It has previously been suggested that siblings may compete directly through begging for a fixed level of provisioning, or that siblings may cooperate in their begging in order to jointly elevate the level of provisioning by adults. We studied the begging behaviour of meerkat Suricata suricatta pups, explored how it changed as the begging behaviour of their littermates altered, and asked how the adults responded to group-level changes in begging. We found conflicting evidence for classic models of competitive and cooperative begging. Pups reared in larger litters begged at higher rates, yet experimentally increasing begging levels within groups caused individual begging rates to decrease. Pups decreased begging rates when close to other begging pups, and pups spaced further apart were fed more. Adults increased their overall level of provisioning as group levels of begging increased, but per capita provisioning decreased. Adults preferred to provision speakers playing back recordings of two pups begging alternately to recordings of the same two pups begging simultaneously. Therefore, we suggest that meerkat pups avoid some of the costs of direct competition imposed by an escalation of begging as other pups beg, by begging in gaps between the bouts of others or avoiding littermates. Such behaviour is also preferred by provisioning adults, thus providing additional benefits to the pups. PMID- 19129140 TI - Development of the five primary podia from the coeloms of a sea star larva: homology with the echinoid echinoderms and other deuterostomes. AB - Confocal laser scanning microscopy of larvae of the asteroid Parvulastra exigua was used to investigate the development of the five primary podia from the coeloms in the echinoderm phylum in an approach to the problem of morphological homology in the deuterostome phyla. The development is shown from an early brachiolaria larval stage to a pre-settlement late brachiolaria larval stage. In the early brachiolaria larva, a single enterocoele connected to the archenteron has formed into two lateral coeloms and an anterior coelom. The primary podia form from the coelomic regions on the left side of the brachiolaria larva, while on the right the coelomic regions connect with the exterior through the pore canal and hydropore. The anterior coelom forms the coelom of the brachia. Homology between the primary podia of the asteroid and the echinoid classes of echinoderms is described and extended to coeloms of other deuterostome phyla. PMID- 19129141 TI - Lophotrochozoa internal phylogeny: new insights from an up-to-date analysis of nuclear ribosomal genes. AB - Resolving the relationships among animal phyla is a key biological problem that remains to be solved. Morphology is unable to determine the relationships among most phyla and although molecular data have unveiled a new evolutionary scenario, they have their own limitations. Nuclear ribosomal genes (18S and 28S rDNA) have been used effectively for many years. However, they are considered of limited use for resolving deep divergences such as the origin of the bilaterians, due to certain drawbacks such as the long-branch attraction (LBA) problem. Here, we attempt to overcome these pitfalls by combining several methods suggested in previous studies and routinely used in contemporary standard phylogenetic analyses but that have not yet been applied to any bilaterian phylogeny based on these genes. The methods used include maximum likelihood and Bayesian inference, the application of models with rate heterogeneity across sites, wide taxon sampling and compartmentalized analyses for each problematic clade. The results obtained show that the combination of the above-mentioned methodologies minimizes the LBA effect, and a new Lophotrochozoa phylogeny emerges. Also, the Acoela and Nemertodermatida are confirmed with maximum support as the first branching bilaterians. Ribosomal RNA genes are thus a reliable source for the study of deep divergences in the metazoan tree, provided that the data are treated carefully. PMID- 19129142 TI - Individual experience and evolutionary history of predation affect expression of heritable variation in fish personality and morphology. AB - Predation plays a central role in evolutionary processes, but little is known about how predators affect the expression of heritable variation, restricting our ability to predict evolutionary effects of predation. We reared families of three spined stickleback Gasterosteus aculeatus from two populations-one with a history of fish predation (predator sympatric) and one without (predator naive)-and experimentally manipulated experience of predators during ontogeny. For a suite of ecologically relevant behavioural ('personality') and morphological traits, we then estimated two key variance components, additive genetic variance (VA) and residual variance (VR), that jointly shape narrow-sense heritability (h2=VA/(VA+VR)). Both population and treatment differentially affected VA versus VR, hence h2, but only for certain traits. The predator-naive population generally had lower VA and h2 values than the predator-sympatric population for personality behaviours, but not morphological traits. Values of VR and h2 were increased for some, but decreased for other personality traits in the predator exposed treatment. For some personality traits, VA and h2 values were affected by treatment in the predator-naive population, but not in the predator-sympatric population, implying that the latter harboured less genetic variation for behavioural plasticity. Replication and experimental manipulation of predation regime are now needed to confirm that these population differences were related to variation in predator-induced selection. Cross-environment genetic correlations (rA) were tight for most traits, suggesting that predator-induced selection can affect the evolution of the same trait expressed in the absence of predators. The treatment effects on variance components imply that predators can affect evolution, not only by acting directly as selective agents, but also by influencing the expression of heritable variation. PMID- 19129143 TI - Significant effects of Pgi genotype and body reserves on lifespan in the Glanville fritillary butterfly. AB - Individuals with a particular variant of the gene phosphoglucose isomerase (Pgi) have been shown to have superior dispersal capacity and fecundity in the Glanville fritillary butterfly (Melitaea cinxia), raising questions about the mechanisms that maintain polymorphism in this gene in the field. Here, we investigate how variation in the Pgi genotype affects female and male life history under controlled conditions. The most striking effect is the longer lifespan of genotypes with high dispersal capacity, especially in non-reproducing females. Butterflies use body reserves for somatic maintenance and reproduction, but different resources (in thorax versus abdomen) are used under dissimilar conditions, with some interactions with the Pgi genotype. These results indicate life-history trade-offs that involve resource allocation and genotypexenvironment interactions, and these trade-offs are likely to contribute to the maintenance of Pgi polymorphism in the natural populations. PMID- 19129144 TI - Facialmetric similarities mediate mate choice: sexual imprinting on opposite-sex parents. PMID- 19129145 TI - [Travel thrombosis, 2008]. AB - In Hungary, the first studies on travel thrombosis were published at the beginning of the 2000s. In this paper recent investigational results of this special type of venous thrombosis have been reviewed. Travel thrombosis is a subgroup of sitting thromboses. It is a consequence of prolonged sitting which is common of ground transportation and air travel. More and more computer-linked sitting thromboses have been observed as well. Long-haul air travel related venous thrombosis is a multifactorial disease. Possible contributory risk factors are in connection with the milieu of the cabin. Various investigations evaluated the effect of immobilization and hypobaric hypoxia on thrombin generation and fibrinolysis. The studies differed much in participants' characteristics, duration and type of exposure and statistical analysis, so the results are contradictory. Personal, traveller-related risk factors may be regarded as triggers. The presently available evidences do not permit to assess the exact actual risk. For healthy young passengers there seem to be low risk. However, passengers suffering from predisposing factors for venous thromboembolism can be exposed to serious hazards, if they fly more than 5000 km or travel more than 8 hours. Proper safety measures are summarized on the basis of recent international recommendations. PMID- 19129146 TI - [Characteristics of the main groups of human host-defensive peptides]. AB - In humans the three main groups of antimicrobial peptides are the defensins, the cathelicidins and the histatins. They differ widely in their biochemical properties and in the spectrum of their antimicrobial activities. For quite a while they were regarded only as new-type antimicrobial agents. Recent studies revealed, however, that functions of these peptides extend far beyond their antimicrobial activities. They were shown to be implicated in a remarkably broad range of other - likewise host defence related - biological processes. They proved to be important components of the innate immune system. Furthermore it was also shown that they interact with various receptors on the immature dendritic cells and lymphocytes resulting in the activation of adaptive immune responses, in which they were shown to play further immunomodulatory roles, too. Pertaining LL-37 it has even been proposed that it has more potent immunomodulatory activities than antimicrobial function. Human alpha-defensins were shown to be active across species in mice and to have immunoadjuvant effects. Recently numerous papers have been reported on studies providing abundant evidences that several diseases in humans are characterized by impairment in the function of these small host defensive peptides. The recognition of the multifunctional role of these peptides further raised the interest of the pharmaceutical industry toward them. PMID- 19129147 TI - [Detection and molecular epidemiology of respiratory syncytial virus type A and B strains in childhood respiratory infections in Hungary]. AB - Human respiratory syncytial virus (hRSV) is one of the major causes of respiratory infection of infants and children worldwide. The molecular epidemiology of hRSV is unknown in Hungary. AIMS: Our aims were the molecular detection and genetic analysis of hRSV from childhood respiratory infections in Hungary. MATERIALS AND METHODS: Nasopharyngeal aspirates were collected from children under the age of 10 years with acute respiratory infections provided by the Pediatric Department of the Hospital for Chest Diseases in Mosdos. Samples were taken from 15 October to 15 May in seasons of 2005/2006 and 2006/2007. The clinical and epidemiological data were collected prospectively. The amplification of the surface fusion glycoprotein (F) and the attachment glycoprotein (G) genes of viral RNA was made by RT-PCR method. PCR-products were sequenced and analyzed by phylogenetic analysis. RESULTS: Nasopharyngeal aspirates of 104 children were examined out of which 23 (22.1%) samples - 16 males (69.6%) and 7 females (30.4%) - (first season: 1/49, 2%; second season: 22/55, 40%) contained hRSV. The hRSV infections were taking place from December to March. The average age was 2.1 years (1 month to 8 years). The leading symptoms were dropping nose, fever, cough and wheezing. Thirty-nine point one percent of the hRSV infected children had underlying disease. Based upon the F region 22 (96%), viruses genetically belonged to type A and 1 (4%) was classified as type B hRSV. Based upon the G region, out of the 11 type A viruses 8 (72.7%) belonged to group GA5 and 3 (27.3%) to group GA2. Viral nucleotide sequence was identical in several cases. CONCLUSIONS: To our knowledge, this is the first report on molecular detection and genetic analysis of the two types (A and B) of hRSV of children under the age of 10 with respiratory infections in Hungary. In winter and spring hRSV is an important cause of childhood respiratory infections, particularly in infants, often requiring hospitalization. PMID- 19129148 TI - [Surgical treatment of acute type-B aortic dissection associated with cocaine use]. AB - Cocaine abuse is on a rise in Hungary as well. It is known that cocaine users have a higher risk developing cardiovascular complications, for example aortic dissection. Almost all patients in Hungary suffering from type B aortic dissection are referred to our department for treatment. AIM: We introduce the case of a regular cocaine user, who suffered an acute type B aortic dissection and was treated surgically. To our best knowledge this is the first similar case in our country to be published. METHOD: Case presentation. RESULTS: We performed a successful operation: acute thoracoabdominal aortic refenestration, no complication was detected. The patient is doing well three months after the procedure, returned to his regular activities, he is normotensively receiving medical treatment, and he gave up cocaine. CONCLUSIONS: Thoracoabdominal aortic refenestration can save the life of patients presenting with acute type B dissection. Good long-term result depends on adequate hypertension control and cocaine abstinence. As the frequency of cocaine abuse increases in Hungary, similar cases may be more often encountered. PMID- 19129149 TI - [Erectile dysfunctions]. PMID- 19129153 TI - Lung cancer deaths from indoor radon and the cost effectiveness and potential of policies to reduce them. AB - OBJECTIVE: To determine the number of deaths from lung cancer related to radon in the home and to explore the cost effectiveness of alternative policies to control indoor radon and their potential to reduce lung cancer mortality. DESIGN: Cost effectiveness analysis. SETTING: United Kingdom. DATA SOURCES: Epidemiological data on risks from indoor radon and from smoking, vital statistics on deaths from lung cancer, survey information on effectiveness and costs of radon prevention and remediation. MAIN OUTCOME MEASURES: Estimated number of deaths from lung cancer related to indoor radon, lifetime risks of death from lung cancer before and after various potential interventions to control radon, the cost per quality adjusted life year (QALY) gained from different policies for control of radon, and the potential of those policies to reduce lung cancer mortality. RESULTS: The mean radon concentration in UK homes is 21 becquerels per cubic metre (Bq/m(3)). Each year around 1100 deaths from lung cancer (3.3% of all deaths from lung cancer) are related to radon in the home. Over 85% of these arise from radon concentrations below 100 Bq/m(3) and most are caused jointly by radon and active smoking. Current policy requiring basic measures to prevent radon in new homes in selected areas is highly cost effective, and such measures would remain cost effective if extended to the entire UK, with a cost per QALY gained of pound11,400 ( euro12 200; $16,913). Current policy identifying and remediating existing homes with high radon levels is, however, neither cost effective (cost per QALY gained pound36,800) nor effective in reducing lung cancer mortality. CONCLUSIONS: Policies requiring basic preventive measures against radon in all new homes throughout the UK would be cost effective and could complement existing policies to reduce smoking. Policies involving remedial work on existing homes with high radon levels cannot prevent most radon related deaths, as these are caused by moderate exposure in many homes. These conclusions are likely to apply to most developed countries, many with higher mean radon concentrations than the UK. PMID- 19129154 TI - Indoor radon and deaths from lung cancer. PMID- 19129155 TI - Natural frequencies help older adults and people with low numeracy to evaluate medical screening tests. AB - BACKGROUND: Understanding information about medical screening tests often requires estimating positive predictive values (i.e., posterior probabilities), which is a notoriously difficult task. Previous studies have shown that representation of information in terms of natural frequencies (i.e., counts of occurrences that preserve base rates) facilitates judgments of positive predictive values. The objective of this study was to investigate whether natural frequencies facilitate accurate estimates in elderly people and whether performance depends on numeracy skills. Elderly people are more often than younger people required to use such information to make informed choices regarding medical procedures (e.g., screenings). METHOD: This was an experimental study in which information about 2 medical screening tests was presented either as conditional probabilities or natural frequencies. Participants were 47 older adults (62-77 years of age; average numeracy score 8.6) and 115 younger adults (18-35 years of age; average numeracy score 10.3). RESULTS: When the screening information was presented in terms of conditional probabilities, only 15% of the younger adults and 18% of the older adults provided accurate estimates in at least 1 of the tasks. When information was presented in terms of natural frequencies, 55% of the younger adults and 58% of the elderly participants gave correct estimates. This effect occurred without explicit training. Furthermore, participants with higher numeracy scores performed better in the estimation tasks than those with lower numeracy scores. CONCLUSIONS: Natural frequencies help elderly and young patients-including those with lower numeracy skills-to understand positive predictive values of medical screening tests. PMID- 19129156 TI - A hybrid cohort individual sampling natural history model of age-related macular degeneration: assessing the cost-effectiveness of screening using probabilistic calibration. AB - BACKGROUND: Age-related macular degeneration (AMD) is a leading cause of visual impairment and blindness. It is likely that treatment of AMD at earlier stages is more effective than later treatment; thus, screening for AMD should be considered. The aim of this study was to develop a natural history model of AMD to estimate the cost-effectiveness of screening. METHODS: A hybrid cohort/individual sampling decision analytic model was developed. Primary data sets, expert elicitation, and data from the literature were used to populate the model. To incorporate joint parameter uncertainty, and to populate unobservable parameters, an innovative form of probabilistic calibration was applied to a range of output parameters. RESULTS: In the reference case, annual screening from age 60 y is the most cost-effective option, although this is subject to high levels of uncertainty. Alternative, age-specific utility values show that screening is predicted to be less cost-effective, assuming interventions that reduce progression to wet AMD moderately improve the cost-effectiveness of screening, whereas the addition of anti-vascular endothelial growth factor therapy for juxtafoveal or subfoveal wet AMD lesions improves the cost effectiveness of screening significantly. CONCLUSIONS: The extent of the uncertainty around the mean results, and the additional resources and possible reorganization of services required to implement screening, indicate that it may be preferable to reduce the level of uncertainty before implementing screening for AMD. Initial actions may be best targeted at assessing how routine data may be used to describe clinical presentation, a screening pilot study, and a secondary costing study. PMID- 19129157 TI - Cost-effectiveness of insuring the uninsured: the case of Korean American children. AB - BACKGROUND: More than one-quarter of Korean American children are uninsured, and many are eligible for children's health insurance programs. The objective of this study is to evaluate the effectiveness and cost-effectiveness of different school based health insurance strategies to provide coverage to uninsured Korean American children. METHOD: The authors used a quasi-experimental nonequivalent control group design, conducted from July to December 2005 in Los Angeles, California. The subjects were in 3 groups/schools (n = 1181). Parents received a variation of outreach methods (i.e., information sheets, school site presentations, automated telephone messages, personal telephone calls) and application assistance (i.e., telephone helpline, on-site assistance). The authors used bivariate and multivariable analysis to assess effectiveness. Cost effectiveness was performed using a 3-stage model and Monte Carlo simulation. RESULTS: Of the uninsured in the intensive intervention group, 41% applied for insurance compared with 13% of the control group (P = 0:002). This success was due to personal telephone calls. Of the uninsured in the intensive intervention group, 23% enrolled in insurance compared with 10% of the control group (P = 0:048). The most common reason reported by parents for nonenrollment despite assistance was failure to mail in the application. Despite the small increase in insurance enrollment from the intensive intervention strategy compared with the control group, it is cost-effective. CONCLUSIONS: Personal telephone calls are effective in increasing outreach for children's health insurance application. However, more research is needed to investigate why parents fail to mail in the application. Ultimately, insuring more children will result in cost-effective improvements in quality-adjusted life years. PMID- 19129158 TI - Intrinsic non-symbiotic and truncated haemoglobins and heterologous Vitreoscilla haemoglobin expression in plants. AB - To date, haemoglobins (Hbs) have been shown to exist in all kingdoms of life. The least studied and understood groups are plant non-symbiotic haemoglobins (nsHbs) and the recently found plant truncated Hbs (trHbs). From a biotechnological point of view, the best characterized and almost exclusively applied Hb is the bacterial Vitreoscilla haemoglobin (VHb). In this review, the present state of knowledge of structural features and ligand binding kinetics of plant nsHbs and trHbs and their proposed roles as oxygen carriers, oxygen sensors, and for oxygen storage, in nitric oxide (NO) detoxification, and in peroxidase activity are described. Furthermore, in order to predict the functioning of plant Hbs, their characteristics will be compared with those of the better known bacterial globins. In this context, the effects of heterologous applications of VHb on plants are reviewed. Finally, the challenging future of plant Hb research is discussed. PMID- 19129159 TI - Changes in the effective gravitational field strength affect the state of phosphorylation of stress-related proteins in callus cultures of Arabidopsis thaliana. AB - In a recent study it was shown that callus cell cultures of Arabidopsis thaliana respond to changes in gravitational field strengths by changes in protein expression. Using ESI-MS/MS for proteins with differential abundance after separation by 2D-PAGE, 28 spots which changed reproducibly and significantly in amount (P <0.05) after 2 h of hypergravity (18 up-regulated, 10 down-regulated) could be identified. The corresponding proteins were largely involved in stress responses, including the detoxification of reactive oxygen species (ROS). In the present study, these investigations are extended to phosphorylated proteins. For this purpose, callus cell cultures of Arabidopsis thaliana were exposed to hypergravity (8 g) and simulated weightlessness (random positioning; RP) for up to 30 min, a period of time which yielded the most reliable data. The first changes, however, were visible as early as 10 min after the start of treatment. In comparison to 1 g controls, exposure to hypergravity resulted in 18 protein spots, and random positioning in 25, respectively, with increased/decreased signal intensity by at least 2-fold (P <0.05). Only one spot (alanine aminotransferase) responded the same way under both treatments. After 30 min of RP, four spots appeared, which could not be detected in control samples. Among the protein spots altered in phosphorylation, it was possible to identify 24 from those responding to random positioning and 12 which responded to 8 g. These 12 proteins (8 g) are partly (5 out of 12) the same as those changed in expression after exposure to 2 h of hypergravity. The respective proteins are involved in scavenging and detoxification of ROS (32%), primary metabolism (20.5%), general signalling (14.7%), protein translation and proteolysis (14.7%), and ion homeostasis (8.8%). Together with our recent data on protein expression, it is assumed that changes in gravitational fields induce the production of ROS. Our data further indicate that responses toward RP are more by post-translational protein modulation (most changes in the degree of phosphorylation occur under RP treatment) than by protein expression (hypergravity). PMID- 19129160 TI - Expression profiling of ascorbic acid-related genes during tomato fruit development and ripening and in response to stress conditions. AB - L-ascorbate (the reduced form of vitamin C) participates in diverse biological processes including pathogen defence mechanisms, and the modulation of plant growth and morphology, and also acts as an enzyme cofactor and redox status indicator. One of its chief biological functions is as an antioxidant. L ascorbate intake has been implicated in the prevention/alleviation of varied human ailments and diseases including cancer. To study the regulation of accumulation of this important nutraceutical in fruit, the expression of 24 tomato (Solanum lycopersicon) genes involved in the biosynthesis, oxidation, and recycling of L-ascorbate during the development and ripening of fruit have been characterized. Taken together with L-ascorbate abundance data, the results show distinct changes in the expression profiles for these genes, implicating them in nodal regulatory roles during the process of L-ascorbate accumulation in tomato fruit. The expression of these genes was further studied in the context of abiotic and post-harvest stress, including the effects of heat, cold, wounding, oxygen supply, and ethylene. Important aspects of the hypoxic and post-anoxic response in tomato fruit are discussed. The data suggest that L-galactose-1 phosphate phosphatase could play an important role in regulating ascorbic acid accumulation during tomato fruit development and ripening. PMID- 19129162 TI - Allantoate amidohydrolase transcript expression is independent of drought tolerance in soybean. AB - Drought is a limiting factor for N(2) fixation in soybean [Glycine max (L.) Merr.] thereby resulting in reduced biomass accumulation and yield. Drought sensitive genotypes accumulate ureides, a product of N(2) fixation, during drought stress; however, drought-tolerant genotypes have lower shoot ureide concentrations, which appear to alleviate drought stress on N(2) fixation. A key enzyme involved in ureide breakdown in shoots is allantoate amidohydrolase (AAH). It is hypothesized that AAH gene expression in soybean determines shoot ureide concentrations during water-deficit stress and is responsible for the differential sensitivities of the N(2)-fixation response to drought among soybean genotypes. The objectives were to examine the relationship between AAH transcript levels and shoot ureide concentration and drought tolerance. Drought-tolerant (Jackson) and drought-sensitive (Williams) genotypes were subjected to three water-availability treatments: well-watered control, moderate water-deficit stress, and severe water-deficit stress. Shoot ureide concentrations were examined, in addition to gene expression of AAH and DREB2, a gene expressed during water-deficit stress. As expected, DREB2 expression was detected only during severe water-deficit stress, and shoot ureide concentrations were greatest in the drought-sensitive genotype relative to the drought-tolerant genotype during water-deficit stress. However, expression of AAH transcripts was similar among water treatments and genotypes, indicating that AAH mRNA was not closely associated with drought tolerance. Ureide concentrations in shoots were weakly associated with AAH mRNA levels. These results indicate that AAH expression is probably not associated with the increased ureide catabolism observed in drought tolerant genotypes, such as Jackson. Further study of AAH at the post translational and enzymatic levels is warranted in order to dissect the potential role of this gene in drought tolerance. PMID- 19129161 TI - Tobacco Arp3 is localized to actin-nucleating sites in vivo. AB - The polarity of actin is a central determinant of intracellular transport in plant cells. To visualize actin polarity in living plant cells, the tobacco homologue of the actin-related protein 3 (ARP3) was cloned and a fusion with the red fluorescent protein (RFP) was generated. Upon transient expression of these fusions in the tobacco cell line BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2), punctate structures were observed near the nuclear envelope and in the cortical plasma. These dots could be shown to decorate actin filaments by expressing RFP ARP3 in a marker line, where actin was tagged by GFP (green fluorescent protein) FABD (fimbrin actin-binding domain 2). When actin filaments were disrupted by latrunculin B or by prolonged cold treatment, and subsequently allowed to recover, the actin filaments reformed from the RFP-ARP3 structures, that therefore represented actin nucleation sites. The intracellular distribution of these sites was followed during the formation of pluricellular files, and it was observed that the density of RFP-ARP3 increased in the apex of the polarized, terminal cells of a file, whereas it was equally distributed in the central cells of a file. These findings are interpreted in terms of position-dependent differences of actin organization. PMID- 19129163 TI - Structural insights into glycoside hydrolase family 32 and 68 enzymes: functional implications. AB - Glycoside hydrolases (GH) have been shown to play unique roles in various biological processes like the biosynthesis of glycans, cell wall metabolism, plant defence, signalling, and the mobilization of storage reserves. To date, GH are divided into more than 100 families based upon their overall structure. GH32 and GH68 are combined in clan GH-J, not only harbouring typical hydrolases but also non-Leloir type transferases (fructosyltransferases), involved in fructan biosynthesis. This review summarizes the recent structure-function research progress on plant GH32 enzymes, and highlights the similarities and differences compared with the microbial GH32 and GH68 enzymes. A profound analysis of ligand bound structures and site-directed mutagenesis experiments identified key residues in substrate (or inhibitor) binding and recognition. In particular, sucrose can bind as inhibitor in Cichorium intybus 1-FEH IIa, whereas it binds as substrate in Bacillus subtilis levansucrase and Arabidopsis thaliana cell wall invertase (AtcwINV1). In plant GH32, a single residue, the equivalent of Asp239 in AtcwINV1, appears to be important for sucrose stabilization in the active site and essential in determining sucrose donor specificity. PMID- 19129164 TI - A 64 kDa sucrose binding protein is membrane-associated and tonoplast-localized in developing mung bean seeds. AB - Sucrose binding proteins (SBPs) were predicted to be membrane-associated, but have been shown to localize in the lumen of protein storage vacuoles of various seeds. In this study, a new 64 kDa SBP has been identified from developing mung bean (Vigna radiata) seeds (here termed VrSBP1) via MS/MS analysis and N-terminal amino acid sequencing analysis and specific antibodies were generated using purified VrSBP1 proteins. Western blot analysis with the new VrSBP1 antibodies showed that, similar to most seed storage proteins, VrSBP1 proteins accumulated during seed development and were subsequently mobilized once the mung bean seeds germinated. Immunogold electron microscope (EM) studies on ultra-thin sections of high-pressure freezing/frozen substituted developing mung bean cotyledons demonstrated that VrSBP1 was localized specifically to the tonoplast of the protein storage vacuole and to the limiting membrane of a novel putative prevacuolar compartment. Biochemical and subcellular fractionation studies further demonstrated that VrSBP1 proteins were membrane-associated in developing mung beans, consistent with their tonoplast localization. This study thus shows convincing evidence of tonoplast-localization of a plant SBP for its future functional characterization and provides a model of studying non-integral membrane proteins associated with the tonoplasts in plant cells. PMID- 19129165 TI - Gene expression studies in kiwifruit and gene over-expression in Arabidopsis indicates that GDP-L-galactose guanyltransferase is a major control point of vitamin C biosynthesis. AB - Vitamin C (L-ascorbic acid, AsA) is an essential metabolite for plants and animals. Kiwifruit (Actinidia spp.) are a rich dietary source of AsA for humans. To understand AsA biosynthesis in kiwifruit, AsA levels and the relative expression of genes putatively involved in AsA biosynthesis, regeneration, and transport were correlated by quantitative polymerase chain reaction in leaves and during fruit development in four kiwifruit genotypes (three species; A. eriantha, A. chinensis, and A. deliciosa). During fruit development, fruit AsA concentration peaked between 4 and 6 weeks after anthesis with A. eriantha having 3-16-fold higher AsA than other genotypes. The rise in AsA concentration typically occurred close to the peak in expression of the L-galactose pathway biosynthetic genes, particularly the GDP-L-galactose guanyltransferase gene. The high concentration of AsA found in the fruit of A. eriantha is probably due to higher expression of the GDP-mannose-3',5'-epimerase and GDP-L-galactose guanyltransferase genes. Over-expression of the kiwifruit GDP-L-galactose guanyltransferase gene in Arabidopsis resulted in up to a 4-fold increase in AsA, while up to a 7-fold increase in AsA was observed in transient expression studies where both GDP-L-galactose guanyltransferase and GDP-mannose-3',5'-epimerase genes were co-expressed. These studies show the importance of GDP-L-galactose guanyltransferase as a rate-limiting step to AsA, and demonstrate how AsA can be significantly increased in plants. PMID- 19129166 TI - Architectural phenotypes in the transparent testa mutants of Arabidopsis thaliana. AB - Flavonoids are low molecular weight secondary plant metabolites with a myriad of functions. As flavonoids affect auxin transport (an important growth-controlling hormone) and are biologically active in eukaryotes, flavonoid mutants were expected to have undescribed architectural phenotypes. The Arabidopsis thaliana transparent testa (tt) mutants are compromised in the enzymatic steps or transcriptional regulators affecting flavonoid synthesis. tt mutant seedlings were grown on hard-slanted agar (a stress condition), under varying light conditions, and in soil to examine the resulting growth patterns. These tt mutants revealed a wide variety of architectural phenotypes in root and aerial tissues. Mutants with increased inflorescences, siliques, and lateral root density or reduced stature are traits that could affect plant yield or performance under certain environmental conditions. The regulatory genes affected in architectural traits may provide useful molecular targets for examination in other plants. PMID- 19129167 TI - Occurrence, phylogeny, structure, and function of catalases and peroxidases in cyanobacteria. AB - Cyanobacteria have evolved approximately 3x10(9) years ago from ancient phototrophic microorganisms that already lived on our planet Earth. By opening the era of an aerobic, oxygen-containing biosphere, they are the true pacemakers of geological and biological evolution. Cyanobacteria must have been among the first organisms to elaborate mechanisms for the detoxification of partially reduced oxygen species including (hydrogen) peroxide. Since there is still an suprising lack of knowledge on the type, role, and mechanism(s) of peroxide degrading enzymes in these bacteria, all 44 fully or partially sequenced genomes for haem and non-haem catalases and peroxidases have been critically analysed based on well known structure-function relationships of the corresponding oxidoreductases. It is demonstrated that H(2)O(2)-dismutating enzymes are mainly represented by bifunctional (haem) catalase-peroxidases and (binuclear) manganese catalases, with the latter being almost exclusively found in diazotrophic species. Several strains even lack a gene that encodes an enzyme with catalase activity. Two groups of peroxidases are found. Genes encoding putative (primordial) haem peroxidases (with homology to corresponding mammalian enzymes) and vanadium-containing iodoperoxidases are found only in a few species, whereas genes encoding peroxiredoxins (1-Cys, 2-Cys, type II, and Q-type) are ubiquitous in cyanobacteria. In addition, approximately 70% contain NADPH-dependent glutathione peroxidase-like proteins. The occurrence and phylogeny of these enzymes is discussed, as well as the present knowledge of their physiological role(s). PMID- 19129168 TI - A green fluorescent protein fused to rice prolamin forms protein body-like structures in transgenic rice. AB - Prolamins, a group of rice (Oryza sativa) seed storage proteins, are synthesized on the rough endoplasmic reticulum (ER) and deposited in ER-derived type I protein bodies (PB-Is) in rice endosperm cells. The accumulation mechanism of prolamins, which do not possess the well-known ER retention signal, remains unclear. In order to elucidate whether the accumulation of prolamin in the ER requires seed-specific factors, the subcellular localization of the constitutively expressed green fluorescent protein fused to prolamin (prolamin GFP) was examined in seeds, leaves, and roots of transgenic rice plants. The prolamin-GFP fusion proteins accumulated not only in the seeds but also in the leaves and roots. Microscopic observation of GFP fluorescence and immunocytochemical analysis revealed that prolamin-GFP fusion proteins specifically accumulated in PB-Is in the endosperm, whereas they were deposited in the electron-dense structures in the leaves and roots. The ER chaperone BiP was detected in the structures in the leaves and roots. The results show that the aggregation of prolamin-GFP fusion proteins does not depend on the tissues, suggesting that the prolamin-GFP fusion proteins accumulate in the ER by forming into aggregates. The findings bear out the importance of the assembly of prolamin molecules and the interaction of prolamin with BiP in the formation of ER-derived PBs. PMID- 19129169 TI - Post-veraison sunlight exposure induces MYB-mediated transcriptional regulation of anthocyanin and flavonol synthesis in berry skins of Vitis vinifera. AB - Anthocyanins, flavan-3-ols, and flavonols are the three major classes of flavonoid compounds found in grape berry tissues. Several viticultural practices increase flavonoid content in the fruit, but the underlying genetic mechanisms responsible for these changes have not been completely deciphered. The impact of post-veraison sunlight exposure on anthocyanin and flavonol accumulation in grape berry skin and its relation to the expression of different transcriptional regulators known to be involved in flavonoid synthesis was studied. Treatments consisting of removing or moving aside the basal leaves which shade berry clusters were applied. Shading did not affect sugar accumulation or gene expression of HEXOSE TRANSPORTER 1, although in the leaf removal treatment, these events were retarded during the first weeks of ripening. Flavonols were the most drastically reduced flavonoids following shading and leaf removal treatments, related to the reduced expression of FLAVONOL SYNTHASE 4 and its putative transcriptional regulator MYB12. Anthocyanin accumulation and the expression of CHS2, LDOX, OMT, UFGT, MYBA1, and MYB5a genes were also affected. Other regulatory genes were less affected or not affected at all by these treatments. Non-transcriptional control mechanisms for flavonoid synthesis are also suggested, especially during the initial stages of ripening. Although berries from the leaf removal treatment received more light than shaded fruits, malvidin 3-glucoside and total flavonol content was reduced compared with the treatment without leaf removal. This work reveals that flavonol-related gene expression responds rapidly to field changes in light levels, as shown by the treatment in which shaded fruits were exposed to light in the late stages of ripening. Taken together, this study establishes MYB-specific responsiveness for the effect of sun exposure and sugar transport on flavonoid synthesis. PMID- 19129170 TI - The nature and efficiency of the word reading strategies of orally raised deaf students. AB - The main objective of this study was to unveil similarities and differences in the word reading strategies of orally raised individuals with prelingual deafness and hearing individuals. Relevant data were gathered by a computerized research paradigm asking participants to make rapid same/different judgments for words. There were three distinct study conditions: (a) a visual condition manipulating the visual-perceptional properties of the target word pairs, (b) a phonological condition manipulating their phonological properties, and (c) a control condition. Participants were 31 high school and postgraduate students with prelingual deafness and 59 hearing students (the control group). Analysis of response latencies and accuracy in the three study conditions suggests that the word reading strategies the groups relied upon to process the stimulus materials were of the same nature. Evidence further suggests that prelingual deafness does not undermine the efficiency with which readers use these strategies. To gain a broader understanding of the obtained evidence, participants' performance in the word processing experiment was correlated with their phonemic awareness-the hypothesized hallmark of proficient word reading-and their reading comprehension skills. Findings are discussed with reference to a reading theory that assigns phonology a central role in proficient word reading. PMID- 19129171 TI - Suppression of GFAP toxicity by alphaB-crystallin in mouse models of Alexander disease. AB - Alexander disease (AxD) is a primary disorder of astrocytes caused by dominant mutations in the gene for glial fibrillary acidic protein (GFAP). These mutations lead to protein aggregation and formation of Rosenthal fibers, complex astrocytic inclusions that contain GFAP, vimentin, plectin, ubiquitin, Hsp27 and alphaB crystallin. The small heat shock protein alphaB-crystallin (Cryab) regulates GFAP assembly, and elevation of Cryab is a consistent feature of AxD; however, its role in Rosenthal fibers and AxD pathology is not known. Here, we show in AxD mouse models that loss of Cryab results in increased mortality, whereas elevation of Cryab rescues animals from terminal seizures. When mice with Rosenthal fibers induced by over-expression of GFAP are crossed into a Cryab-null background, over half die at 1 month of age. Restoration of Cryab expression through the GFAP promoter reverses this outcome, showing the effect is astrocyte-specific. Conversely, in mice engineered to express both AxD-associated mutations and elevated GFAP, which despite natural induction of Cryab also die at 1 month, transgenic over-expression of Cryab results in a markedly reduced CNS stress response, restores expression of the glutamate transporter Glt1 (EAAT2) and protects these animals from death. In its most common form, AxD is a devastating neurodegenerative disease, with early onset, characterized by seizures, spasticity and developmental delays, ultimately leading to death. Cryab plays a critical role in tempering AxD pathology and should be investigated as a therapeutic target for this and other diseases with astropathology. PMID- 19129173 TI - Incomplete penetrance and phenotypic variability characterize Gdf6-attributable oculo-skeletal phenotypes. AB - Proteins of the bone morphogenetic protein (BMP) family are known to have a role in ocular and skeletal development; however, because of their widespread expression and functional redundancy, less progress has been made identifying the roles of individual BMPs in human disease. We identified seven heterozygous mutations in growth differentiation factor 6 (GDF6), a member of the BMP family, in patients with both ocular and vertebral anomalies, characterized their effects with a SOX9-reporter assay and western analysis, and demonstrated comparable phenotypes in model organisms with reduced Gdf6 function. We observed a spectrum of ocular and skeletal anomalies in morphant zebrafish, the latter encompassing defective tail formation and altered expression of somite markers noggin1 and noggin2. Gdf6(+/-) mice exhibited variable ocular phenotypes compatible with phenotypes observed in patients and zebrafish. Key differences evident between patients and animal models included pleiotropic effects, variable expressivity and incomplete penetrance. These data establish the important role of this determinant in ocular and vertebral development, demonstrate the complex genetic inheritance of these phenotypes, and further understanding of BMP function and its contributions to human disease. PMID- 19129172 TI - The loss of the snoRNP chaperone Nopp140 from Cajal bodies of patient fibroblasts correlates with the severity of spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a common autosomal recessive neurodegenerative disease caused by reduced survival motor neuron (SMN) levels. The assembly machinery containing SMN is implicated in the biogenesis of the spliceosomal small nuclear ribonucleoproteins (snRNPs). SMN is present in both the cytoplasm and nucleus, where it transiently accumulates in subnuclear domains named Cajal bodies (CBs) and functions in the maturation of snRNPs and small nucleolar (sno)RNPs. The impact of lowering SMN levels on the composition of CBs in SMA cells is still not completely understood. Here, we analyse the CB composition in immortalized and primary fibroblasts from SMA patients. We show that the U snRNA export factors PHAX and chromosome region maintenance 1 and the box C/D snoRNP core protein fibrillarin concentrate in CBs from SMA cells, whereas the box H/ACA core proteins GAR1 and NAP57/dyskerin show reduced CB localization. Remarkably, the functional deficiency in SMA cells is associated with decreased localization of the snoRNP chaperone Nopp140 in CBs that correlates with disease severity. Indeed, RNA interference knockdown experiments in control fibroblasts demonstrate that SMN is required for accumulation of Nopp140 in CBs. Conversely, overexpression of SMN in SMA cells restores the CB localization of Nopp140, whereas SMN mutants found in SMA patients are defective in promoting the association of Nopp140 with CBs. Taken together, we demonstrate that only a subset of CB functions (as indicated by the association of representative factors) are impaired in SMA cells and, importantly, we identify the decrease of Nopp140 localization in CBs as a phenotypic marker for SMA. PMID- 19129174 TI - Evaluation of imputation-based association in and around the integrin-alpha-M (ITGAM) gene and replication of robust association between a non-synonymous functional variant within ITGAM and systemic lupus erythematosus (SLE). AB - We recently identified a novel non-synonymous variant, rs1143679, at exon 3 of the ITGAM gene associated with systemic lupus erythematosus (SLE) susceptibility in European-Americans (EAs) and African-Americans. Using genome-wide association approach, three other studies also independently reported an association between SLE susceptibility and ITGAM or ITGAM-ITGAX region. The primary objectives of this study are to assess whether single or multiple causal variants from the same gene or any nearby gene(s) are involved in SLE susceptibility and to confirm a robust ITGAM association across nine independent data sets (n = 8211). First, we confirmed our previously reported association of rs1143679 (risk allele 'A') with SLE in EAs (P = 1.0 x 10(-8)) and Hispanic-Americans (P = 2.9 x 10(-5)). Secondly, using a comprehensive imputation-based association test, we found that ITGAM is one of the major non-human leukocyte antigen susceptibility genes for SLE, and the strongest association for EA is the same coding variant rs1143679 (log(10)Bayes factor=20, P = 6.17 x 10(-24)). Thirdly, we determined the robustness of rs1143679 association with SLE across three additional case-control samples, including UK (P = 6.2 x 10(-8)), Colombian (P = 3.6 x 10(-7)), Mexican (P = 0.002), as well as two independent sets of trios from UK (P(TDT) = 1.4 x 10( 5)) and Mexico (P(TDT) = 0.015). A meta-analysis combing all independent data sets greatly reinforces the association (P(meta) = 7.1 x 10(-50), odds ratio = 1.83, 95% confidence interval = 1.69-1.98, n = 10 046). However, this ITGAM association was not observed in the Korean or Japanese samples, in which rs1143679 is monomorphic for the non-risk allele (G). Taken together along with our earlier findings, these results demonstrate that the coding variant, rs1143679, best explains the ITGAM-SLE association, especially in European- and African-derived populations, but not in Asian populations. PMID- 19129175 TI - Calcium signaling in brain mitochondria: interplay of malate aspartate NADH shuttle and calcium uniporter/mitochondrial dehydrogenase pathways. AB - Ca2+ signaling in mitochondria has been mainly attributed to Ca2+ entry to the matrix through the Ca2+ uniporter and activation of mitochondrial matrix dehydrogenases. However, mitochondria can also sense increases in cytosolic Ca2+ through a mechanism that involves the aspartate-glutamate carriers, extramitochondrial Ca2+ activation of the NADH malate-aspartate shuttle (MAS). Both pathways are linked through the shared substrate alpha-ketoglutarate (alphaKG). Here we have studied the interplay between the two pathways under conditions of Ca2+ activation. We show that alphaKG becomes limiting when Ca2+ enters in brain or heart mitochondria, but not liver mitochondria, resulting in a drop in alphaKG efflux through the oxoglutarate carrier and in a drop in MAS activity. Inhibition of alphaKG efflux and MAS activity by matrix Ca2+ in brain mitochondria was fully reversible upon Ca2+ efflux. Because of their differences in cytosolic calcium concentration requirements, the MAS and Ca2+ uniporter mitochondrial dehydrogenase pathways are probably sequentially activated during a Ca2+ transient, and the inhibition of MAS at the center of the transient may provide an explanation for part of the increase in lactate observed in the stimulated brain in vivo. PMID- 19129177 TI - A conserved hydrophobic tetrad near the C terminus of the secretory Na+-K+-2Cl- cotransporter (NKCC1) is required for its correct intracellular processing. AB - Little is known about the intracellular folding and trafficking of integral membrane proteins. Here we identify a hydrophobic amino acid tetrad (ILLV) close to the C terminus of the secretory Na+-K+-2Cl- cotransporter (NKCC1) that is important for the proper intracellular processing of this protein. This tetrad appears in a C-terminal sequence pattern that is conserved across species in a number of members of the NKCC1 gene family (slc12) of electroneutral salt transporters. We studied the effects of various mutations of these amino acids on NKCC1 transiently transfected into HEK-293 cells. Our results show that mutation of two of these residues to alanine leads to a >50% reduction in expression and complex glycosylation levels and that multiple mutations to alanine have cumulative effects. By contrast, scrambling of these amino acids, or mutation of other nearby conserved C-terminal residues, has little effect on these parameters. Mutation of ILLV to AAAA reduces complex glycosylation of NKCC1 by approximately 90% and results in a protein that does not form stable dimers and is retained in the endoplasmic reticulum in a highly aggregated state. Our results are consistent with the hypothesis that mutation of the hydrophobic tetrad ILLV to AAAA leads to the ab initio misfolding and concomitant aggregation of this NKCC1 mutant, resulting in its retention in the endoplasmic reticulum. PMID- 19129176 TI - Solution structure of the NaV1.2 C-terminal EF-hand domain. AB - Voltage-gated sodium channels initiate the rapid upstroke of action potentials in many excitable tissues. Mutations within intracellular C-terminal sequences of specific channels underlie a diverse set of channelopathies, including cardiac arrhythmias and epilepsy syndromes. The three-dimensional structure of the C terminal residues 1777-1882 of the human NaV1.2 voltage-gated sodium channel has been determined in solution by NMR spectroscopy at pH 7.4 and 290.5 K. The ordered structure extends from residues Leu-1790 to Glu-1868 and is composed of four alpha-helices separated by two short anti-parallel beta-strands; a less well defined helical region extends from residue Ser-1869 to Arg-1882, and a disordered N-terminal region encompasses residues 1777-1789. Although the structure has the overall architecture of a paired EF-hand domain, the NaV1.2 C terminal domain does not bind Ca2+ through the canonical EF-hand loops, as evidenced by monitoring 1H,15N chemical shifts during aCa2+ titration. Backbone chemical shift resonance assignments and Ca2+ titration also were performed for the NaV1.5 (1773-1878) isoform, demonstrating similar secondary structure architecture and the absence of Ca2+ binding by the EF-hand loops. Clinically significant mutations identified in the C-terminal region of NaV1 sodium channels cluster in the helix I-IV interface and the helix II-III interhelical segment or in helices III and IV of the NaV1.2 (1777-1882) structure. PMID- 19129178 TI - Phospholipid transfer protein Sec14 is required for trafficking from endosomes and regulates distinct trans-Golgi export pathways. AB - A protein known to regulate both lipid metabolism and vesicular transport is the phosphatidylcholine/phosphatidylinositol transfer protein Sec14 of Saccharomyces cerevisiae. Sec14 is thought to globally affect secretion from the trans-Golgi. The results from a synthetic genetic array screen for genes whose inactivation impaired growth of cells with a temperature-sensitive SEC14 allele implied Sec14 regulates transport into and out of the Golgi. This prompted us to examine the role of Sec14 in various vesicular transport pathways. We determined that Sec14 function was required for the route followed by Bgl2, whereas trafficking of other secreted proteins, including Hsp150, Cts1, Scw4, Scw10, Exg1, Cis3, and Ygp1, still occurred, indicating Sec14 regulates specific trans-Golgi export pathways. Upon diminution of Sec14 function, the v-SNARE Snc1 accumulated in endosomes and the trans-Golgi. Its accumulation in endosomes is consistent with Sec14 being required for transport from endosomes to the trans-Golgi. Sec14 was also required for trafficking of Ste3 and the lipophilic dye FM4-64 from the plasma membrane to the vacuole at the level of the endosome. The combined genetic and cell biology data are consistent with regulation of endosome trafficking being a major role for Sec14. We further determined that lipid ligand occupancy differentially regulates Sec14 functions. PMID- 19129179 TI - Anionic phospholipids lose their procoagulant properties when incorporated into high density lipoproteins. AB - Blood coagulation involves a series of enzymatic protein complexes that assemble on the surface of anionic phospholipid. To investigate whether apolipoproteins affect coagulation reactions, they were included during the preparation of anionic phospholipid vesicles using a detergent solubilization-dialysis method. Apolipoprotein components of high density lipoproteins, especially apolipoprotein A-I, had a pronounced anticoagulant effect. The anionic phospholipids lost their procoagulant effect when the vesicle preparation method was performed in the presence of apolipoprotein A-I. The anionic phospholipid-apolipoprotein A-I particles were 8-10 nm in diameter and contained around 60-80 phospholipid molecules, depending on the phospholipid composition. The phospholipids of these particles were unable to support the activation of prothrombin by factor Xa in the presence of factor Va and unable to support binding of factor Va, whereas binding of prothrombin and factor Xa were efficient. Phospholipid transfer protein was shown to mediate transfer of phospholipids from liposomes to apolipoprotein A-I-containing reconstituted high density lipoprotein. In addition, serum was also shown to neutralize the procoagulant effect of anionic liposomes and to efficiently mediate transfer of phospholipids from liposomes to either apolipoprotein A-I- or apolipoprotein B-containing particles. In conclusion, apolipoprotein A-I was found to neutralize the procoagulant properties of anionic phospholipids by arranging the phospholipids in surface areas that are too small to accommodate the prothrombinase complex. This anionic phospholipid scavenger function may be an important mechanism to control the exposure of such phospholipids to circulating blood and thereby prevent inappropriate stimulation of blood coagulation. PMID- 19129181 TI - Platelet factor 4 impairs the anticoagulant activity of activated protein C. AB - Platelet factor 4 (PF4) is an abundant platelet alpha-granule chemokine released following platelet activation. PF4 interacts with thrombomodulin and the gamma carboxyglutamic acid (Gla) domain of protein C, thereby enhancing activated protein C (APC) generation by the thrombin-thrombomodulin complex. However, the protein C Gla domain not only mediates protein C activation in vivo, but also plays a critical role in modulating the diverse functional properties of APC once generated. In this study we demonstrate that PF4 significantly inhibits APC anti coagulant activity. PF4 inhibited both protein S-dependent APC anticoagulant function in plasma and protein S-dependent factor Va (FVa) proteolysis 3- to 5 fold, demonstrating that PF4 impairs protein S cofactor enhancement of APC anticoagulant function. Using recombinant factor Va variants FVa-R506Q/R679Q and FVa-R306Q/R679Q, PF4 was shown to impair APC proteolysis of FVa at position Arg(306) by 3-fold both in the presence and absence of protein S. These data suggest that PF4 contributes to the poorly understood APC resistance phenotype associated with activated platelets. Finally, despite PF4 binding to the APC Gla domain, we show that APC in the presence of PF4 retains its ability to initiate PAR-1-mediated cytoprotective signaling. In summary, we propose that PF4 acts as a critical regulator of APC generation, but also differentially targets APC toward cytoprotective, rather than anticoagulant function at sites of vascular injury with concurrent platelet activation. PMID- 19129180 TI - Crystal structure of Streptococcus pyogenes sortase A: implications for sortase mechanism. AB - Sortases are a family of Gram-positive bacterial transpeptidases that anchor secreted proteins to bacterial cell surfaces. These include many proteins that play critical roles in the virulence of Gram-positive bacterial pathogens such that sortases are attractive targets for development of novel antimicrobial agents. All Gram-positive pathogens express a "housekeeping" sortase that recognizes the majority of secreted proteins containing an LPXTG wall-sorting motif and covalently attaches these to bacterial cell wall peptidoglycan. Many Gram-positive pathogens also express additional sortases that link a small number of proteins, often with variant wall-sorting motifs, to either other surface proteins or peptidoglycan. To better understand the mechanisms of catalysis and substrate recognition by the housekeeping sortase produced by the important human pathogen Streptococcus pyogenes, the crystal structure of this protein has been solved and its transpeptidase activity established in vitro. The structure reveals a novel arrangement of key catalytic residues in the active site of a sortase, the first that is consistent with kinetic analysis. The structure also provides a complete description of residue positions surrounding the active site, overcoming the limitation of localized disorder in previous structures of sortase A-type proteins. Modification of the active site Cys through oxidation to its sulfenic acid form or by an alkylating reagent supports a role for a reactive thiol/thiolate in the catalytic mechanism. These new insights into sortase structure and function could have important consequences for inhibitor design. PMID- 19129182 TI - Genetic suppressors and recovery of repressed biochemical memory. PMID- 19129183 TI - The Kaposi's Sarcoma-associated Herpesvirus-encoded vIRF-3 Inhibits Cellular IRF 5. AB - Kaposi's sarcoma-associated herpesvirus encodes four genes with homology to the family of interferon regulatory factors (IRFs). At least one of these viral IRFs, vIRF-3, is expressed in latently Kaposi's sarcoma-associated herpesvirus-infected primary effusion lymphoma (PEL) cells and is essential for the survival of PEL cells. We now report that vIRF-3 interacts with cellular IRF-5, thereby inhibiting binding of IRF-5 to interferon-responsive promoter elements. Consequently, vIRF-3 blocked IRF-5-mediated promoter activation. A central double helix motif present in vIRF-3 was sufficient to abrogate both DNA binding and transcriptional transactivation by IRF-5. Upon DNA damage or activation of the interferon or Toll-like receptor pathways, cytoplasmic IRF-5 has been reported to be translocated to the nucleus, which results in induction of both p53 independent apoptosis and p21-mediated cell cycle arrest. We report here that IRF 5 is present in the nuclei of PEL cells without interferon stimulation. Silencing of vIRF-3 expression in PEL cells was accompanied by increased sensitivity to interferon-mediated apoptosis and up-regulation of IRF-5 target genes. In addition, vIRF-3 antagonized IRF-5-mediated activation of the p21 promoter. The data presented here indicate that vIRF-3 contributes to immune evasion and sustained proliferation of PEL cells by releasing IRF-5 from transcription complexes. PMID- 19129184 TI - Epidermal growth factor-like repeats of thrombospondins activate phospholipase Cgamma and increase epithelial cell migration through indirect epidermal growth factor receptor activation. AB - Thrombospondin (TSP) 1 is a trimeric multidomain protein that contains motifs that recognize distinct host cell receptors coupled to multiple signaling pathways. Selected TSP1-induced cellular responses are tyrosine kinase-dependent, and TSP1 contains epidermal growth factor (EGF)-like repeats. Specific receptor interactions or functions for the EGF-like repeats have not been identified. We asked whether one or more biological responses to TSP1 might be explained through EGF receptor (EGFR) activation. In A431 cells, TSP1 increased autophosphorylation of Tyr-1068 of EGFR in a dose- and time-dependent manner. The ability of TSP1 to activate EGFR was replicated by the tandem EGF-like repeats as a recombinant protein. The three EGF-like repeats alone produced a high level of Tyr-1068 phosphorylation. EGF-like repeats from TSP2 and TSP4 also activated EGFR. Tyr 1068 phosphorylation was less when individual EGF-like repeats were tested or flanking sequences were added to the three EGF-like repeats. TSP1 and its EGF like repeats also increased phosphorylation of EGFR Tyr-845, Tyr-992, Tyr-1045, Tyr-1086, and Tyr-1173, activated phospholipase Cgamma, and increased cell migration. No evidence was found for binding of the EGF-like repeats to EGFR. Instead, EGFR activation in response to TSP1 or its EGF-like repeats required matrix metalloprotease activity, including activity of matrix metalloprotease 9. Access to the ligand-binding portion of the EGFR ectodomain was also required. These findings suggest release of an endogenous EGFR ligand in response to ligation of a second unknown receptor by the TSPs. PMID- 19129185 TI - Biochemical and structural analysis of bacterial O-antigen chain length regulator proteins reveals a conserved quaternary structure. AB - Lipopolysaccharide (LPS) is a major component of the Gram-negative outer membrane and is an important virulence determinant. The O-antigen polysaccharide of the LPS molecule provides protection from host defenses, and the length of O-antigen chains plays a pivotal role. In the Wzy-dependent O-antigen biosynthesis pathway, the integral inner membrane protein Wzz determines the O-antigen chain length. How these proteins function is currently unknown, but the hypothesis includes activities such as a "molecular ruler" or a "molecular stopwatch," and other possibilities may exist. Wzz homologs are membrane proteins with two transmembrane helices that flank a large periplasmic domain. Recent x-ray crystallographic studies of the periplasmic portions of Wzz proteins found multiple oligomeric forms, with quaternary structures favoring the "molecular ruler" interpretation. Here, we have studied full-length Wzz proteins with the transmembrane portions embedded in lipid membranes. Using electron microscopy and image analysis we find a unique hexameric state rather than differing oligomeric forms. The data suggest that in vivo Wzz proteins determine O-antigen chain length via subtle structure-function relationships at the level of primary, secondary, or tertiary structure within the context of a hexameric complex. PMID- 19129186 TI - The secondary multidrug/proton antiporter MdfA tolerates displacements of an essential negatively charged side chain. AB - The largest family of solute transporters includes ion motive force-driven secondary transporters. Several well characterized solute-specific transport systems in this group have at least one irreplaceable acidic residue that plays a critical role in energy coupling during transport. Previous studies have established the importance of acidic residues in substrate recognition by major facilitator superfamily secondary multidrug transporters, but their role in the transport mechanism remained unknown. We have been investigating the involvement of acidic residues in the mechanism of MdfA, an Escherichia coli secondary multidrug/proton antiporter. We demonstrated that no single negatively charged side chain plays an irreplaceable role in MdfA. Accordingly, we hypothesized that MdfA might be able to utilize at least two acidic residues alternatively. In this study, we present evidence that indeed, unlike solute-specific secondary transporters, MdfA tolerates displacements of an essential negative charge to various locations in the putative drug translocation pathway. The results suggest that MdfA utilizes a proton translocation strategy that is less sensitive to perturbations in the geometry of the proton-binding site, further illustrating the exceptional structural promiscuity of multidrug transporters. PMID- 19129187 TI - An iron-sulfur cluster is essential for the binding of broken DNA by AddAB-type helicase-nucleases. AB - The bacterial helicase-nuclease complex AddAB converts double-stranded DNA breaks into substrates for RecA-dependent recombinational repair. Here we show that the AddB subunit contains a novel class of nuclease domain distinguished by the presence of an iron-sulfur cluster. The cluster is coordinated by an unusual arrangement of cysteine residues that originate from both sides of the AddB nuclease, forming an "iron staple" that is required for the local structural integrity of this domain. Disruption of the iron-sulfur cluster by mutagenesis eliminates the ability of AddAB to bind to duplex DNA ends without affecting the single-stranded DNA-dependent ATPase activity. Sequence analysis suggests that a related iron staple nuclease domain is present in the eukaryotic DNA replication/repair factor Dna2, where it is also associated with a DNA helicase motor. PMID- 19129189 TI - The Drosophila melanogaster Gene cg4930 Encodes a High Affinity Inhibitor for Endonuclease G. AB - Endonuclease G (EndoG) is a mitochondrial enzyme believed to be released during apoptosis to participate in the degradation of nuclear DNA. This paper describes a Drosophila protein, EndoGI, which inhibits EndoG specifically. EndoG and EndoGI associate with subpicomolar affinity, forming a 2:1 complex in which dimeric EndoG is bound by two tandemly repeated homologous domains of monomeric EndoGI. Binding appears to involve the active site of EndoG. EndoGI is present in the cell nucleus at micromolar concentrations. Upon induction of apoptosis, levels of the inhibitor appear to be reduced, and it is relocalized to the cytoplasm. EndoGI, encoded by the predicted open reading frame cg4930, is expressed throughout Drosophila development. Flies homozygous for a hypomorphic EndoGI mutation have a strongly reduced viability, which is modulated by genetic background and diet. We propose that EndoGI protects the cell against low levels of EndoG outside mitochondria. PMID- 19129188 TI - Occupancy and functional architecture of the pigment binding sites of photosystem II antenna complex Lhcb5. AB - Lhcb5 is an antenna protein that is highly conserved in plants and green algae. It is part of the inner layer of photosystem II antenna system retained in high light acclimated plants. To study the structure-function relation and the role of individual pigments in this complex, we (i) "knocked out" each of the chromophores bound to multiple (nine total) chlorophyll sites and (ii) exchanged the xanthophylls bound to the three xanthophyll sites. The occupancy and associated energy of the pigment binding sites were determined. The role of the individual pigments in protein folding, stability, energy transfer, and dissipation was studied in vitro. The results indicate that lutein has a primary role in the folding and stability of the complex, whereas violaxanthin and zeaxanthin have a negative effect on folding yield and stability, respectively. The data showed a distinct function for the L1 and L2 carotenoid binding sites, the former preferentially involved in gathering the excitation energy to chlorophyll a (Chl a), whereas the latter modulates the concentration of chlorophyll singlet excited states dependent on the xanthophylls bound to it, likely via an interaction with Chl-603. Our results also underscored the role of zeaxanthin and lutein in quenching the excitation energy, whereas violaxanthin was shown to be very effective in energy transfer. The characteristics of the isolated proteins were consistent with the observed role of Lhcb5 in vivo in catalyzing fluorescence quenching upon zeaxanthin binding. PMID- 19129190 TI - The Listeria monocytogenes sortase-B recognizes varied amino acids at position 2 of the sorting motif. AB - Sortases are bacterial enzymes that anchor surface proteins covalently to the peptidoglycan upon cleavage of a motif located at their C-terminal end. Motifs recognized by sortases of the class-B (SrtB) are defined by the consensus sequence NP(Q/K)(T/S)(N/G/S)(D/A). Evidence supporting this consensus is limited to IsdC of Staphylococcus aureus and Bacillus anthracis, cleaved at motifs NPQTN and NPKTG, respectively. In Listeria monocytogenes, StrB has two substrates, Lmo2185 and Lmo2186, containing NAKTN and NKVTN (or the overlapping sequence NPKSS) as putative sorting motifs. Some of these motifs do not match the consensus, because they lack either proline (P) at position 2 or glutamine/lysine (Q/K) at position 3. Here, we identified NPKSS as a sorting motif of Lmo2186 by monitoring anchoring to peptidoglycan of chimeras lacking each of its two predicted motifs. Motif-swapping experiments confirmed that NPKSS, but not NKVTN, could replace NAKTN for anchoring of an Lmo2185 chimera. Residue substitutions in the NPKSS sequence revealed the essentiality of proline at position 2 for recognition of this particular motif. Lysine at position 3 was however dispensable. Deletion of NAKTN, on the other hand, abrogated SrtB-mediated anchoring of the Lmo2185 chimera. NAKTN, therefore, represents an exception to the rule of a conserved proline in position 2 of the sorting motif. Taken together, our data indicate that proline is not absolutely required for substrate recognition by sortases of the class-B. In addition, they prove the capacity of a single sortase, as SrtB of L. monocytogenes, to recognize varied amino acids at position 2 of the sorting motif. PMID- 19129192 TI - Beta-sheet-dependent dimerization is essential for the stability of NhaA Na+/H+ antiporter. AB - A structural model of the NhaA dimer showed that a beta-hairpin of each monomer combines to form a beta-sheet at the periplasmic side of the membrane. By Cys scanning the entire beta-hairpin and testing each Cys replacement for functionality and intermolecular cross-linking, we found that Gln47 and Arg49 are critical for the NhaA dimer and that K57C causes an acidic shift of 1 pH unit to the pH dependence of NhaA. Comparing the growth of the NhaA variants with the previously isolated beta-hairpin deleted mutant (Delta(P45-N58)) and the wild type validated that NhaA dimers have an advantage over monomers in growth under extreme stress conditions and unraveled that during this growth the apparent Km for Na+ of Delta(P45-N58) was increased 50-fold as compared with the wild type. Remarkably, the effect of the extreme stress on the NhaA variants is reversible. Testing the temperature stability (4-55 degrees C) of the NhaA variants in dodecyl maltoside micells showed that the mutants impaired in dimerization were much less temperature-stable than the wild type. We suggest that NhaA dimers are crucial for the stability of the antiporter under extreme stress conditions. PMID- 19129191 TI - The 48-kDa alternative translation isoform of PP2A:B56epsilon is required for Wnt signaling during midbrain-hindbrain boundary formation. AB - Alternative translation is an underappreciated post-transcriptional regulation mechanism. Although only a small number of genes are found to be alternatively translated, most genes undergoing alternative translation play important roles in tumorigenesis and development. Protein phosphatase 2A (PP2A) is involved in many cellular events during tumorigenesis and development. The specificity, localization, and activity of PP2A are regulated by B regulatory subunits. B56epsilon, a member of the B56 regulatory subunit family, is involved in multiple signaling pathways and regulates a number of developmental processes. Here we report that B56epsilon is alternatively translated, leading to the production of a full-length form and a shorter isoform that lacks the N-terminal 76 amino acid residues of the full-length form. Alternative translation of B56epsilon occurs through a cap-dependent mechanism. We provide evidence that the shorter isoform is required for Wnt signaling and regulates the midbrain/hindbrain boundary formation during Xenopus embryonic development. This demonstrates that the shorter isoform of B56epsilon has important biological functions. Furthermore, we show that the N-terminal sequence of B56epsilon, which is not present in the shorter isoform, contains a nuclear localization signal, whereas the C terminus of B56epsilon contains a nuclear export signal. The shorter isoform, which lacks the N-terminal nuclear localization signal, is restricted to the cytoplasm. In contrast, the full-length form can be localized to the nucleus in a cell type-specific manner. The finding that B56epsilon is alternatively translated adds a new level of regulation to PP2A holoenzymes. PMID- 19129193 TI - Identification of adjacent binding sites for the YY1 and E4BP4 transcription factors in the ovine PrP (Prion) gene promoter. AB - The PrP gene encodes the cellular isoform of the prion protein (PrP(c)) which has been shown to be crucial to the development of transmissible spongiform encephalopathies (TSEs). PrP knock-out mice, which do not express endogenous PrP(c), exhibit resistance to TSE disease. The regulation of PrP gene expression represents, therefore, a crucial factor in the development of TSEs. Two sequence motifs in the PrP promoter (positions -287 to -263 from transcriptional start) were previously reported as being highly conserved, and it was suggested that they represent binding sites for as yet unidentified transcription factors. To test this hypothesis, binding of nuclear proteins was analyzed by electrophoretic mobility shift assays using ovine or murine cells and tissues with radiolabeled DNA probes containing the conserved motif sequences. Specific binding was observed to both motifs, and polymorphic variants of these motifs exhibited differential binding. Two proteins bound to these motifs were identified as the Yin Yang 1 (YY1) (motif 1) and E4BP4 (motif 2) transcription factors. Functional promoter analysis of four different promoter variants revealed that motif 1 (YY1) was associated with inhibitory activity in the context of the PrP promoter, whereas motif 2 (E4BP4) was linked to a slight enhancing activity. This represents the first demonstration of binding of nuclear factors to two highly conserved DNA sequence motifs within mammalian PrP promoters. The action of these factors on the PrP promoter is haplotype-specific, leading us to propose that the prion protein expression pattern and, with it, the distribution of TSE infectivity may be associated with PrP promoter genotype. PMID- 19129194 TI - In Vitro and in Vivo Characterization of Molecular Interactions between Calmodulin, Ezrin/Radixin/Moesin, and L-selectin. AB - L-selectin is a cell adhesion molecule that tethers leukocytes to the luminal walls of venules during inflammation and enables them to roll under the force of blood flow. Clustering of L-selectin during rolling is thought to promote outside in signals that lead to integrin activation and chemokine receptor expression, ultimately contributing to leukocyte arrest. Several studies have underscored the importance of the L-selectin cytoplasmic tail in functionally regulating adhesion and signaling. Interestingly, the L-selectin tail comprises only 17 amino acids, and yet it is thought to bind simultaneously to several proteins. For example, constitutive association of calmodulin (CaM) and ezrin/radixin/moesin (ERM) to L selectin confers resistance to proteolysis and microvillar positioning, respectively. In this report we found that recombinant purified CaM and ERM bound non-competitively to the same tail of L-selectin. Furthermore, molecular modeling supported the possibility that CaM, L-selectin, and moesin could form a heterotrimeric complex. Finally, using fluorescence lifetime imaging microscopy to measure fluorescence resonance energy transfer, it was shown that CaM, L selectin, and ERM could interact simultaneously in vivo. Moreover, L-selectin clustering promoted CaM/ERM interaction in cis (i.e. derived from neighboring L selectin tails). These results highlight a novel intracellular event that occurs as a consequence of L-selectin clustering, which could participate in transducing signals that promote the transition from rolling to arrest. PMID- 19129195 TI - A novel protein acts as a negative regulator of prophenoloxidase activation and melanization in the freshwater crayfish Pacifastacus leniusculus. AB - Melanization is an important immune component of the innate immune system of invertebrates and is vital for defense as well as for wound healing. In most invertebrates melanin synthesis is achieved by the prophenoloxidase-activating system, a proteolytic cascade similar to vertebrate complement. Even though melanin formation is necessary for host defense in crustaceans and insects, the process needs to be tightly regulated because of the hazard to the animal of unwanted production of quinone intermediates and melanization in places where it is not suitable. In the present study we have identified a new melanization inhibition protein (MIP) from the hemolymph of the crayfish, Pacifastacus leniusculus. Crayfish MIP has a similar function as the insect MIP molecule we recently discovered in the beetle Tenebrio molitor but interestingly has a completely different sequence. Crayfish MIP as well as Tenebrio MIP do not affect phenoloxidase activity in itself but instead interfere with the melanization reaction from quinone compounds to melanin. Importantly, crayfish MIP in contrast to Tenebrio MIP contains a fibrinogen-like domain, most similar to the substrate recognition domain of vertebrate l-ficolins. Surprisingly, an Asp-rich region similar to that found in ficolins that is likely to be involved in Ca2+ binding is present in crayfish MIP. However, crayfish MIP did not show any hemagglutinating activity as is common for the vertebrate ficolins. A mutant form of MIP with a deletion lacking four Asp amino acids from the Asp-rich region lost most of its activity, implicating that this part of the protein is involved in regulating the prophenoloxidase activating cascade. Overall, a new negative regulator of melanization was identified in freshwater crayfish that shows interesting parallels with proteins (i.e. ficolins) involved in vertebrate immune response. PMID- 19129196 TI - HTLV-1 Tax is a critical lipid raft modulator that hijacks IkappaB kinases to the microdomains for persistent activation of NF-kappaB. AB - Upon T cell activation, IkappaB kinases (IKKs) are transiently recruited to the plasma membrane-associated lipid raft microdomains for activation of NF-kappaB in promoting T cell proliferation. Retroviral Tax proteins from human T cell leukemia virus type 1 and type 2 (HTLV-1 and -2) are capable of activating IKK, yet only HTLV-1 infection causes T cell leukemia, which correlates with persistent activation of NF-kappaB induced by Tax1. Here, we show that the Tax proteins exhibit differential modes of IKK activation. The subunits of IKK are constitutively present in lipid rafts in activated forms in HTLV-1-infected T cells that express Tax. Disruption of lipid rafts impairs IkappaB kinase activation by Tax1. We also show that the cytoplasmic Tax1 protein persistently resides in the Golgi-associated lipid raft microdomains. Tax1 directs lipid raft translocation of IKK through selective interaction with IKKgamma and accordingly, depletion of IKKgamma impairs Tax1-directed lipid raft recruitment of IKKalpha and IKKbeta. In contrast, Tax2 activates NF-kappaB in a manner independent of lipid raft recruitment of IKK. These findings indicate that Tax1 actively recruits IKK to the lipid raft microdomains for persistent activation of NF kappaB, thereby contributing to HTLV-1 oncogenesis. PMID- 19129197 TI - Cooperativity of Mus81.Mms4 with Rad54 in the resolution of recombination and replication intermediates. AB - The Saccharomyces cerevisiae Mus81.Mms4 protein complex, a DNA structure-specific endonuclease, helps preserve genomic integrity by resolving pathological DNA structures that arise from damaged or aborted replication forks and may also play a role in the resolution of DNA intermediates arising through homologous recombination. Previous yeast two-hybrid studies have found an interaction of the Mus81 protein with Rad54, a Swi2/Snf2-like factor that serves multiple roles in homologous recombination processes. However, the functional significance of this novel interaction remains unknown. Here, using highly purified S. cerevisiae proteins, we show that Rad54 strongly stimulates the Mus81.Mms4 nuclease activity on a broad range of DNA substrates. This nuclease enhancement does not require ATP binding nor its hydrolysis by Rad54. We present evidence that Rad54 acts by targeting the Mus81.Mms4 complex to its DNA substrates. In addition, we demonstrate that the Rad54-mediated enhancement of the Mus81.Mms4 (Eme1) nuclease function is evolutionarily conserved. We propose that Mus81.Mms4 together with Rad54 efficiently process perturbed replication forks to promote recovery and may constitute an alternative mechanism to the resolution/dissolution of the recombination intermediates by Sgs1.Top3. These findings provide functional insights into the biological importance of the higher order complex of Mus81.Mms4 or its orthologue with Rad54. PMID- 19129198 TI - Preparation and properties of asymmetric vesicles that mimic cell membranes: effect upon lipid raft formation and transmembrane helix orientation. AB - A methyl-beta-cyclodextrin-induced lipid exchange technique was devised to prepare small unilamellar vesicles with stable asymmetric lipid compositions. Asymmetric vesicles that mimic biological membranes were prepared with sphingomyelin (SM) or SM mixed with 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) as the predominant lipids in the outer leaflet and dioleoylphosphatidylcholine (DOPC), POPC, 1-palmitoyl-2-oleoyl-phosphatidyl-L serine (POPS), or POPS mixed with 1-palmitoyl-2-oleoyl-phosphatidylethanolamine (POPE) in the inner leaflet. Fluorescence-based assays were developed to confirm lipid asymmetry. Cholesterol was introduced into these vesicles using a second methyl-beta-cyclodextrin exchange step. In asymmetric vesicles composed of SM outside, DOPC inside (SMo/DOPCi) or SM outside, 2:1 mol:mol POPE:POPS inside (SMo/2:1 POPE:POPSi) the outer leaflet SM formed an ordered state with a thermal stability similar to that in pure SM vesicles and significantly greater than that in symmetric vesicles with the same overall lipid composition. Analogous behavior was observed in vesicles containing cholesterol. This shows that an asymmetric lipid distribution like that in eukaryotic plasma membranes can be conducive to ordered domain (raft) formation. Furthermore asymmetric vesicles containing approximately 25 mol % cholesterol formed ordered domains more thermally stable than those in asymmetric vesicles lacking cholesterol, showing that the crucial ability of cholesterol to stabilize ordered domain formation is likely to contribute to ordered domain formation in cell membranes. Additional studies demonstrated that hydrophobic helix orientation is affected by lipid asymmetry with asymmetry favoring formation of the transmembrane configuration. The ability to form asymmetric vesicles represents an important improvement in model membrane studies and should find many applications in the future. PMID- 19129199 TI - Functional protein delivery into neurons using polymeric nanoparticles. AB - An efficient route for delivering specific proteins and peptides into neurons could greatly accelerate the development of therapies for various diseases, especially those involving intracellular defects such as Parkinson disease. Here we report the novel use of polybutylcyanoacrylate nanoparticles for delivery of intact, functional proteins into neurons and neuronal cell lines. Uptake of these particles is primarily dependent on endocytosis via the low density lipoprotein receptor. The nanoparticles are rapidly turned over and display minimal toxicity to cultured neurons. Delivery of three different functional cargo proteins is demonstrated. When primary neuronal cultures are treated with recombinant Escherichia coli beta-galactosidase as nanoparticle cargo, persistent enzyme activity is measured beyond the period of nanoparticle degradation. Delivery of the small GTPase rhoG induces neurite outgrowth and differentiation in PC12 cells. Finally, a monoclonal antibody directed against synuclein is capable of interacting with endogenous alpha-synuclein in cultured neurons following delivery via nanoparticles. Polybutylcyanoacrylate nanoparticles are thus useful for intracellular protein delivery in vitro and have potential as carriers of therapeutic proteins for treatment of neuronal disorders in vivo. PMID- 19129201 TI - Prolonged mechanical noise restores tactile sense in diabetic neuropathic patients. AB - Acute application of stochastic resonance (SR), defined as a subsensory level of mechanical noise presented directly to sensory neurons, improves the vibration and tactile perception in diabetic patients with mild to moderate neuropathy. This study examined the effect of 1 hour of continuous SR stimulation on sensory nerve function. Twenty diabetic patients were studied. The effect of stimulation was measured at 2 time points, at the beginning and after 60 minutes of continual SR stimulation. This effect was measured using the vibration perception threshold (VPT) at the big toe under 2 conditions: a null (no SR) condition and active SR, defined as mechanical noise below the subject's own threshold of perception. The measurements under null and active conditions were done randomly and the examiner was blinded regarding the type of condition. Immediately after SR application, the VPT with SR in null condition was similar to baseline (32.2 +/- 13.1, P = nonsignificant) but was significantly lower during active SR (27.4 +/- 11.9) compared with both baseline (P = .018) and off position (P = .045). The 60 minutes VPT with active SR (28.7 +/- 11.1) reached significance comparing the baseline when one outlier was removed from the analysis (P = .031). It may be concluded that SR for a continuous 60-minute period can sustain the VPT improvement in diabetic patients with moderate to severe neuropathy. These results permit the conclusion that there is no short-term adaptation to the stimulation signal. Long-term application of this technique, perhaps in the form of a continually vibrating shoe insert, or insole, may result in sustained improvement of nerve function. PMID- 19129200 TI - Homology with vesicle fusion mediator syntaxin-1a predicts determinants of epimorphin/syntaxin-2 function in mammary epithelial morphogenesis. AB - We have shown that branching morphogenesis of mammary ductal structures requires the action of the morphogen epimorphin/syntaxin-2. Epimorphin, originally identified as an extracellular molecule, is identical to syntaxin-2, an intracellular molecule that is a member of the extensively investigated syntaxin family of proteins that mediate vesicle trafficking. We show here that, although epimorphin/syntaxin-2 is highly homologous to syntaxin-1a, only epimorphin/syntaxin-2 can stimulate mammary branching morphogenesis. We construct a homology model of epimorphin/syntaxin-2 based on the published structure of syntaxin-1a, and we use this model to identify the structural motif responsible for the morphogenic activity. We identify four residues located within the cleft between helices B and C that differ between syntaxin-1a and epimorphin/syntaxin 2; through site-directed mutagenesis of these four amino acids, we confer the properties of epimorphin for cell adhesion, gene activation, and branching morphogenesis onto the inactive syntaxin-1a template. These results provide a dramatic demonstration of the use of structural information about one molecule to define a functional motif of a second molecule that is related at the sequence level but highly divergent functionally. PMID- 19129202 TI - Cell proliferation induction: healing chronic wounds through low-energy pulsed radiofrequency. AB - Chronic lower extremity wounds are a major source of morbidity and health care expenditure across the world. In the last decade, numerous advanced modalities have become available that can help expedite wound healing when standard wound care modalities are unsuccessful. In this case report, the authors present their early experience with one such modality that uses pulsed radiofrequency energy to initiate cell proliferation induction in chronic lower extremity wounds of different etiologies that had failed to respond to standard therapy. PMID- 19129203 TI - Estimation of patients' organ doses and conceptus doses from selected X-ray examinations in two Nigeria X-ray centres. AB - In this study, organ and conceptus doses of patients undergoing chest, abdomen and skull radiograph examinations at two Nigeria X-ray centres, Niger State General Hospital (NGH) and Two-Tees (TTX), are reported. Air kerma was measured, and entrance surface dose (ESD) and half-value layer estimated for each set of tube potential (kV(p)), focus to skin distance and current-time product (mAs) used for each of the patients included in this study. Results show that the mean air kerma in the two centres are similar for the three projections considered in this study. Organ doses ranged from <0.01 to 2.18 mGy in NGH and from <0.01 to 1.29 mGy in TTX for examinations of the abdomen, from <0.01 to 0.20 mGy in NGH and from <0.01 to 0.13 mGy in TTX for examinations of the skull and from <0.01 to 3.90 mGy in NGH and from <0.01 to 1.96 mGy in TTX for examinations of the chest. Generally, no significant difference is seen between the organ doses of male and female patients. In NGH, organ doses are generally greater than those from TTX for the three examinations. The mean ESDs for examinations of the chest postero anterior, abdomen antero-posterior (AP) and skull AP are, respectively, 5.37, 6.28 and 4.24 mGy in NGH, and 5.82, 5.33 and 4.76 mGy in TTX. The ESDs reported in this study, except for examinations of the chest, are generally lower than comparable values published in the literature. Conceptus doses were also estimated for female patients using normalised published conceptus dose data for abdomen examinations. The estimated conceptus doses were >1 mGy even when the conceptus was located 12 cm below the surface of the abdomen. PMID- 19129204 TI - Fenestrated grafts or debranching procedures for complex abdominal aortic aneurysms. AB - Over the past 15 years, endovascular aneurysm repair of abdominal aortic aneurysms has become widely accepted as a means of treating aneurysms located in the infrarenal portion of the aorta. It has been estimated that 30% to 40% of patients with abdominal aortic aneurysms are not candidates for endovascular repair using the current commercially available devices. The primary limitation has been unfavorable anatomy most often associated with the proximal aortic neck. Although the morbidity and mortality of open pararenal or suprarenal aneurysms has improved, many patients will not tolerate open surgery. Therefore, other techniques need to be employed. This article looks at 2 other techniques to treat complex pararenal, juxtarenal, or thoracoabdominal aneurysms, one being the use of fenestrated devices and the other being visceral artery debranching followed by endovascular grafting. Multiple series are reviewed, and the outcomes are analyzed. PMID- 19129205 TI - A phase III randomized, double-blind, placebo-controlled trial of gabapentin in the management of hot flashes in men (N00CB). AB - INTRODUCTION: Hot flashes represent a significant problem in men undergoing androgen deprivation therapy. MATERIALS AND METHODS: Via a prospective, double blind, placebo-controlled clinical trial, men with hot flashes, on a stable androgen deprivation therapy program for prostate cancer, received a placebo or gabapentin at target doses of 300, 600, or 900 mg/day. Hot flash frequencies and severities were recorded daily during a baseline week and for 4 weeks while the patients took the study medication. RESULTS: In the 214 eligible patients who began the study drug on this trial, comparing the fourth treatment week to the baseline week, mean hot flash scores decreased in the placebo group by 4.1 units and in the three increasing dose gabapentin groups by, 3.2, 4.6, and 7.0 units. Comparing the three combined gabapentin arms to the placebo arm did not result in significant hot flash differences. Wilcoxon rank-sum P values for change in hot flash scores and frequencies after 4 weeks of treatment were 0.10 and 0.02, comparing the highest dose gabapentin arm to the placebo arm, respectively. The gabapentin was well tolerated in this trial. CONCLUSION: These results support that gabapentin decreases hot flashes, to a moderate degree, in men with androgen ablation-related vasomotor dysfunction. PMID- 19129206 TI - DichroCalc--circular and linear dichroism online. AB - MOTIVATION: Circular dichroism (CD) is widely used in studies of protein folding. The CD spectrum of a protein can be estimated from its structure alone, using the well-established matrix method. In the last decade, a related spectroscopy, linear dichroism (LD), has been increasingly applied to study the orientation of proteins in solution. However, matrix method calculations of LD spectra have not been presented before. DichroCalc makes both CD and LD calculations available in an easy-to-use fashion. RESULTS: DichroCalc can be used without registration and calculates CD and LD spectra using a variety of matrix method parameters. PDB files can be uploaded as input or retrieved via their PDB code and a Perl-based parser is offered for easy handling of PDB files. AVAILABILITY: http://comp.chem.nottingham.ac.uk/dichrocalc and http://comp.chem.nottingham.ac.uk/parsepdb. PMID- 19129207 TI - A novel method for large tree visualization. AB - SUMMARY: Many genomic and proteomic analyses generate as a result a tree of genes or proteins. These trees are often large (containing tens of thousands of nodes and edges), and need a visualization tool to fully display all the information contained in the tree. Clustering analysis can be performed on these trees to obtain clusters of proteins, and we need an efficient way to visualize the clustering results. We present a novel tree visualization tool to help with such analyses. AVAILABILITY: http://www2.renci.org/~jeff/software/bin/win32/ProteinVis 2.1.6-win32.zip. PMID- 19129208 TI - PanCGH: a genotype-calling algorithm for pangenome CGH data. AB - MOTIVATION: Pangenome arrays contain DNA oligomers targeting several sequenced reference genomes from the same species. In microbiology, these can be employed to investigate the often high genetic variability within a species by comparative genome hybridization (CGH). The biological interpretation of pangenome CGH data depends on the ability to compare strains at a functional level, particularly by comparing the presence or absence of orthologous genes. Due to the high genetic variability, available genotype-calling algorithms can not be applied to pangenome CGH data. RESULTS: We have developed the algorithm PanCGH that incorporates orthology information about genes to predict the presence or absence of orthologous genes in a query organism using CGH arrays that target the genomes of sequenced representatives of a group of microorganisms. PanCGH was tested and applied in the analysis of genetic diversity among 39 Lactococcus lactis strains from three different subspecies (lactis.cremoris, hordniae) and isolated from two different niches (dairy and plant). Clustering of these strains using the presence/absence data of gene orthologs revealed a clear separation between different subspecies and reflected the niche of the strains. PMID- 19129209 TI - bioDBnet: the biological database network. AB - SUMMARY: bioDBnet is an online web resource that provides interconnected access to many types of biological databases. It has integrated many of the most commonly used biological databases and in its current state has 153 database identifiers (nodes) covering all aspects of biology including genes, proteins, pathways and other biological concepts. bioDBnet offers various ways to work with these databases including conversions, extensive database reports, custom navigation and has various tools to enhance the quality of the results. Importantly, the access to bioDBnet is updated regularly, providing access to the most recent releases of each individual database. AVAILABILITY: http://biodbnet.abcc.ncifcrf.gov. PMID- 19129210 TI - Biodiversity informatics: automated approaches for documenting global biodiversity patterns and processes. AB - MOTIVATION: Data about biodiversity have been scattered in different formats in natural history collections, survey reports and the literature. A central challenge for the biodiversity informatics community is to provide the means to share and rapidly synthesize these data and the knowledge they provide us to build an easily accessible, unified global map of biodiversity. Such a map would provide raw and summary data and information on biodiversity and its change across the world at multiple scales. RESULTS: We discuss a series of steps required to create a unified global map of biodiversity. These steps include: building biodiversity repositories; creating scalable species distribution maps; creating flexible, user-programmable pipelines which enable biodiversity assessment; and integrating phylogenetic approaches into biodiversity assessment. We show two case studies that combine phyloinformatic and biodiversity informatic approaches to document large scale biodiversity patterns. The first case study uses data available from the Barcode of Life initiative in order to make species conservation assessment of North American birds taking into account evolutionary uniqueness. The second case study uses full genomes of influenza A available from Genbank to provide an auto-updating documentation of the evolution and geographic spread of these viruses. AVAILABILITY: Both the website for tracking evolution and spread of influenza A and the website for applying phyloinformatics analysis to Barcode of Life data are available as outcomes of case studies (http://biodiversity.colorado.edu). PMID- 19129211 TI - SCARF: maximizing next-generation EST assemblies for evolutionary and population genomic analyses. AB - SUMMARY: Scaffolded and Corrected Assembly of Roche 454 (SCARF) is a next generation sequence assembly tool for evolutionary genomics that is designed especially for assembling 454 EST sequences against high-quality reference sequences from related species. The program was created to knit together 454 contigs that do not assemble during traditional de novo assembly, using a reference sequence library to orient the 454 sequences. AVAILABILITY: SCARF is freely available at http://msbarker.com/software.htm, and is released under the open source GPLv3 license (http://www.opensource.org/licenses/gpl-3.0.html. PMID- 19129212 TI - Prequips--an extensible software platform for integration, visualization and analysis of LC-MS/MS proteomics data. AB - SUMMARY: We describe an integrative software platform, Prequips, for comparative proteomics-based systems biology analysis that: (i) integrates all information generated from mass spectrometry (MS)-based proteomics as well as from basic proteomics data analysis tools, (ii) visualizes such information for various proteomic analyses via graphical interfaces and (iii) links peptide and protein abundances to external tools often used in systems biology studies. AVAILABILITY: http://prequips.sourceforge.net PMID- 19129213 TI - Increased Nrf2 activation in livers from Keap1-knockdown mice increases expression of cytoprotective genes that detoxify electrophiles more than those that detoxify reactive oxygen species. AB - Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor critical for protection against electrophilic and oxidative stress. In a recently engineered mouse with knockdown of kelch-like ECH associated protein 1 (Keap1-kd mice), the cytosolic repressor of Nrf2, there is a 55% decrease in Keap1 mRNA and a 200% increase in Nrf2 protein in liver. Experiments with Nrf2-null mice have demonstrated the effects of a lack of Nrf2. However, little is known about the biological effects of more Nrf2 activation. Accordingly, the hepatic phenotype of Keap1-kd mice, as well as the hepatic mRNA expression of cytoprotective genes were compared among wild-type, Nrf2-null, and Keap1-kd mice. Three distinct patterns of hepatic gene expression were identified among wild-type, Nrf2-null, and Keap1-kd mice. The first pattern encompassed genes that were lower in Nrf2 null mice and considerably higher in Keap1-kd mice than wild-type mice, which included genes mainly responsible for the detoxification and elimination of electrophiles, such as NAD(P)H:quinone oxidoreductase 1 and glutathione-S transferases (Gst), and multidrug resistance-associated proteins. The second pattern encompassed genes that were lower in Nrf2-null mice but not increased in Keap1-kd mice, and included genes, such as epoxide hydrolase-1, UDP glucuronosyltransferases, aldehyde dehydrogenases, as well as genes important in the detoxification of reactive oxygen species, such as superoxide dismutase 1 and 2, catalase, and peroxiredoxin 1. The third pattern encompassed genes that were not different among wild-type, Nrf2-null, and Keap1-kd mice and included genes such as glutathione peroxidase, microsomal Gsts, and uptake transporters. In conclusion, the present study suggests that increased activation of hepatic Nrf2 is more important for the detoxification and elimination of electrophiles than reactive oxygen species. PMID- 19129214 TI - Genome-wide searching with base-pairing kernel functions for noncoding RNAs: computational and expression analysis of snoRNA families in Caenorhabditis elegans. AB - Despite the accumulating research on noncoding RNAs (ncRNAs), it is likely that we are seeing only the tip of the iceberg regarding our understanding of the functions and the regulatory roles served by ncRNAs in cellular metabolism, pathogenesis and host-pathogen interactions. Therefore, more powerful computational and experimental tools for analyzing ncRNAs need to be developed. To this end, we propose novel kernel functions, called base-pairing profile local alignment (BPLA) kernels, for analyzing functional ncRNA sequences using support vector machines (SVMs). We extend the local alignment kernels for amino acid sequences in order to handle RNA sequences by using STRAL's; scoring function, which takes into account sequence similarities as well as upstream and downstream base-pairing probabilities, thus enabling us to model secondary structures of RNA sequences. As a test of the performance of BPLA kernels, we applied our kernels to the problem of discriminating members of an RNA family from nonmembers using SVMs. The results indicated that the discrimination ability of our kernels is stronger than that of other existing methods. Furthermore, we demonstrated the applicability of our kernels to the problem of genome-wide search of snoRNA families in the Caenorhabditis elegans genome, and confirmed that the expression is valid in 14 out of 48 of our predicted candidates by using qRT-PCR. Finally, highly expressed six candidates were identified as the original target regions by DNA sequencing. PMID- 19129215 TI - Transcriptional control by adenovirus E1A conserved region 3 via p300/CBP. AB - The human adenovirus type 5 (HAdV-5) E1A 13S oncoprotein is a potent regulator of gene expression and is used extensively as a model for transcriptional activation. It possesses two independent transcriptional activation domains located in the N-terminus/conserved region (CR) 1 and CR3. The protein acetyltransferase p300 was previously identified by its association with the N terminus/CR1 portion of E1A and this association is required for oncogenic transformation by E1A. We report here that transcriptional activation by 13S E1A is inhibited by co-expression of sub-stoichiometric amounts of the smaller 12S E1A isoform, which lacks CR3. Transcriptional inhibition by E1A 12S maps to the N terminus and correlates with the ability to bind p300/CBP, suggesting that E1A 12S is sequestering this limiting factor from 13S E1A. This is supported by the observation that the repressive effect of E1A 12S is reversed by expression of exogenous p300 or CBP, but not by a CBP mutant lacking actyltransferase activity. Furthermore, we show that transcriptional activation by 13S E1A is greatly reduced by siRNA knockdown of p300 and that CR3 binds p300 independently of the well-characterized N-terminal/CR1-binding site. Importantly, CR3 is also required to recruit p300 to the adenovirus E4 promoter during infection. These results identify a new functionally significant interaction between E1A CR3 and the p300/CBP acetyltransferases, expanding our understanding of the mechanism by which this potent transcriptional activator functions. PMID- 19129216 TI - A versatile non-radioactive assay for DNA methyltransferase activity and DNA binding. AB - We present a simple, non-radioactive assay for DNA methyltransferase activity and DNA binding. As most proteins are studied as GFP fusions in living cells, we used a GFP binding nanobody coupled to agarose beads (GFP nanotrap) for rapid one-step purification. Immobilized GFP fusion proteins were subsequently incubated with different fluorescently labeled DNA substrates. The absolute amounts and molar ratios of GFP fusion proteins and bound DNA substrates were determined by fluorescence spectroscopy. In addition to specific DNA binding of GFP fusion proteins, the enzymatic activity of DNA methyltransferases can also be determined by using suicide DNA substrates. These substrates contain the mechanism-based inhibitor 5-aza-dC and lead to irreversible covalent complex formation. We obtained covalent complexes with mammalian DNA methyltransferase 1 (Dnmt1), which were resistant to competition with non-labeled canonical DNA substrates, allowing differentiation between methyltransferase activity and DNA binding. By comparison, the Dnmt1(C1229W) catalytic site mutant showed DNA-binding activity, but no irreversible covalent complex formation. With this assay, we could also confirm the preference of Dnmt1 for hemimethylated CpG sequences. The rapid optical read-out in a multi-well format and the possibility to test several different substrates in direct competition allow rapid characterization of sequence-specific binding and enzymatic activity. PMID- 19129217 TI - Identification of an inter-transcription factor regulatory network in human hepatoma cells by Matrix RNAi. AB - Transcriptional regulation by transcriptional regulatory factors (TRFs) of their target TRF genes is central to the control of gene expression. To study a static multi-tiered inter-TRF regulatory network in the human hepatoma cells, we have applied a Matrix RNAi approach in which siRNA knockdown and quantitative RT-PCR are used in combination on the same set of TRFs to determine their interdependencies. This approach focusing on several liver-enriched TRF families, each of which consists of structurally homologous members, revealed many significant regulatory relationships. These include the cross-talks between hepatocyte nuclear factors (HNFs) and the other TRF groups such as CCAAT/enhancer binding proteins (CEBPs), retinoic acid receptors (RARs), retinoid receptors (RXRs) and RAR-related orphan receptors (RORs), which play key regulatory functions in human hepatocytes and liver. In addition, various multi-component regulatory motifs, which make up the complex inter-TRF regulatory network, were identified. A large part of the regulatory edges identified by the Matrix RNAi approach could be confirmed by chromatin immunoprecipitation. The resultant significant edges enabled us to depict the inter-TRF TRN forming an apparent regulatory hierarchy of (FOXA1, RXRA) --> TCF1 --> (HNF4A, ONECUT1) --> (RORC, CEBPA) as the main streamline. PMID- 19129218 TI - The Torso signaling pathway modulates a dual transcriptional switch to regulate tailless expression. AB - The Torso (Tor) signaling pathway activates tailless (tll) expression by relieving tll repression. None of the repressors identified so far, such as Capicuo, Groucho and Tramtrack69 (Ttk69), bind to the tor response element (tor RE) or fully elucidate tll repression. In this study, an expanded tll expression pattern was shown in embryos with reduced heat shock factor (hsf) and Trithorax like (Trl) activities. The GAGA factor, GAF encoded by Trl, bound weakly to the tor-RE, and this binding was enhanced by both Hsf and Ttk69. A similar extent of expansion of tll expression was observed in embryos with simultaneous knockdown of hsf, Trl and ttk69 activities, and in embryos with constitutively active Tor. Hsf is a substrate of mitogen-activated protein kinase and S378 is the major phosphorylation site. Phosphorylation converts Hsf from a repressor to an activator that works with GAF to activate tll expression. In conclusion, the GAF/Hsf/Ttk69 complex binding to the tor-RE remodels local chromatin structure to repress tll expression and the Tor signaling pathway activate tll expression by modulating a dual transcriptional switch. PMID- 19129219 TI - A ChIP-chip approach reveals a novel role for transcription factor IRF1 in the DNA damage response. AB - IRF1 is a transcription factor that regulates key processes in the immune system and in tumour suppression. To gain further insight into IRF1's role in these processes, we searched for new target genes by performing chromatin immunoprecipitation coupled to a CpG island microarray (ChIP-chip). Using this approach we identified 202 new IRF1-binding sites with high confidence. Functional categorization of the target genes revealed a surprising cadre of new roles that can be linked to IRF1. One of the major functional categories was the DNA damage response pathway. In order to further validate our findings, we show that IRF1 can regulate the mRNA expression of a number of the DNA damage response genes in our list. In particular, we demonstrate that the mRNA and protein levels of the DNA repair protein BRIP1 [Fanconi anemia gene J (FANC J)] are upregulated after IRF1 over-expression. We also demonstrate that knockdown of IRF1 by siRNA results in loss of BRIP1 expression, abrogation of BRIP1 foci after DNA interstrand crosslink (ICL) damage and hypersensitivity to the DNA crosslinking agent, melphalan; a characteristic phenotype of FANC J cells. Taken together, our data provides a more complete understanding of the regulatory networks controlled by IRF1 and reveals a novel role for IRF1 in regulating the ICL DNA damage response. PMID- 19129220 TI - Oxidation of a single active site suffices for the functional inactivation of the dimeric Bacillus subtilis OhrR repressor in vitro. AB - Bacillus subtilis OhrR is a dimeric repressor that senses organic peroxides and regulates the expression of the OhrA peroxiredoxin. Derepression results from oxidation of an active site cysteine which ultimately results in formation of a mixed disulfide with a low molecular weight thiol, a cyclic sulfenamide, or overoxidation to the sulfinic or sulfonic acids. We expressed a single-chain OhrR (scOhrR) in which the two monomers were connected by a short amino-acid linker. scOhrR variants containing only one active site cysteine were fully functional as repressors and still responded, albeit with reduced efficacy, to organic peroxides in vivo. Biochemical analyses indicate that oxidation at a single active site is sufficient for derepression regardless of the fate of the active site cysteine. scOhrR with only one active site cysteine in the amino-terminal domain is inactivated at rates comparable to wild-type whereas when the active site is in the carboxyl-terminal domain the protein is inactivated much more slowly. The incomplete derepression noted for single active site variants of scOhrR in vivo is consistent with the hypothesis that protein reduction regenerates active repressor and that, in the cell, oxidation of the second active site may also contribute to derepression. PMID- 19129221 TI - The G140S mutation in HIV integrases from raltegravir-resistant patients rescues catalytic defect due to the resistance Q148H mutation. AB - Raltegravir (MK-0518) is the first integrase (IN) inhibitor to be approved by the US FDA and is currently used in clinical treatment of viruses resistant to other antiretroviral compounds. Virological failure of Raltegravir treatment is associated with mutations in the IN gene following two main distinct genetic pathways involving either the N155 or Q148 residue. Importantly, in most cases, an additional mutation at the position G140 is associated with the Q148 pathway. Here, we investigated the viral DNA kinetics for mutants identified in Raltegravir-resistant patients. We found that (i) integration is impaired for Q148H when compared with the wild-type, G140S and G140S/Q148H mutants; and (ii) the N155H and G140S mutations confer lower levels of resistance than the Q148H mutation. We also characterized the corresponding recombinant INs properties. Enzymatic performances closely parallel ex vivo studies. The Q148H mutation 'freezes' IN into a catalytically inactive state. By contrast, the conformational transition converting the inactive form into an active form is rescued by the G140S/Q148H double mutation. In conclusion, the Q148H mutation is responsible for resistance to Raltegravir whereas the G140S mutation increases viral fitness in the G140S/Q148H context. Altogether, these results account for the predominance of G140S/Q148H mutants in clinical trials using Raltegravir. PMID- 19129222 TI - Identification of pregnane-X receptor target genes and coactivator and corepressor binding to promoter elements in human hepatocytes. AB - Chromatin immunoprecipitation (ChIP) studies were conducted in human hepatocytes treated with rifampicin in order to identify new pregnane-X receptor (PXR) target genes. Genes, both previously known to be involved and not known to be involved in drug disposition, with PXR response elements (PXREs) located upstream, within or downstream from their potentially associated genes, were identified. Validation experiments identified several new drug disposition genes with PXR binding sites. Of these, only CYP4F12 demonstrated increased binding in the presence of rifampicin. The role of PXR in the basal and inductive response of CYP4F12 was confirmed in hepatocytes in which PXR was silenced. We also assessed the association of PXR-coactivators and -corepressors with known and newly identified PXREs. Both PXR and the steroid receptor coactivator (SRC-1) were found to bind to PXREs in the absence of rifampicin, although binding was stronger after rifampicin treatment. We observed promoter-dependent patterns with respect to the binding of various coactivators and corepressors involved in the regulation of CYP4F12, CYP3A4, CYP2B6, UGT1A1 and P-glycoprotein. In conclusion, our findings indicate that PXR is involved in the regulation of CYP4F12 and that PXR along with SRC1 binds to a broad range of promoters but that many of these are not inducible by rifampicin. PMID- 19129223 TI - An insulator element 3' to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells. AB - Regulation of expression of the CFTR gene is poorly understood. Elements within the basal promoter of the gene do not fully explain CFTR expression patterns, suggesting that cis-regulatory elements are located elsewhere, either within the locus or in adjacent chromatin. We previously mapped DNase I hypersensitive sites (DHS) in 400 kb spanning the CFTR locus including a cluster of sites close to the 3'-end of the gene. Here we focus on a DHS at +6.8 kb from the CFTR translation end-point to evaluate its potential role in regulating expression of the gene. This DHS, which encompasses a consensus CTCF-binding site, was evident in primary human epididymis cells that express abundant CFTR mRNA. We show by DNase I footprinting and electophoretic mobility shift assays that the cis-regulatory element within this DHS binds CTCF in vitro. We further demonstrate that the element functions as an enhancer blocker in a well-established in vivo assay, and by using chromatin immunoprecipitation that it recruits CTCF in vivo. Moreover, we reveal that in primary epididymis cells, the +6.8 kb DHS interacts closely with the CFTR promoter, suggesting that the CFTR locus exists in a looped conformation, characteristic of an active chromatin hub. PMID- 19129224 TI - Specificity of the ribosomal A site for aminoacyl-tRNAs. AB - Although some experiments suggest that the ribosome displays specificity for the identity of the esterified amino acid of its aminoacyl-tRNA substrate, a study measuring dissociation rates of several misacylated tRNAs containing the GAC anticodon from the A site showed little indication for such specificity. In this article, an expanded set of misacylated tRNAs and two 2'-deoxynucleotide substituted mRNAs are used to demonstrate the presence of a lower threshold in k(off) values for aa-tRNA binding to the A site. When a tRNA binds sufficiently well to reach this threshold, additional stabilizing effects due to the esterified amino acid or changes in tRNA sequence are not observed. However, specificity for different amino acid side chains and the tRNA body is observed when tRNA binding is sufficiently weaker than this threshold. We propose that uniform aa-tRNA binding to the A site may be a consequence of a conformational change in the ribosome, induced by the presence of the appropriate combination of contributions from the anticodon, amino acid and tRNA body. PMID- 19129225 TI - Structural and biochemical characterization of MepR, a multidrug binding transcription regulator of the Staphylococcus aureus multidrug efflux pump MepA. AB - MepR is a multidrug binding transcription regulator that represses expression of the Staphylococcus aureus multidrug efflux pump gene, mepA, as well as its own gene. MepR is induced by multiple cationic toxins, which are also substrates of MepA. In order to understand the gene regulatory and drug-binding mechanisms of MepR, we carried out biochemical, in vivo and structural studies. The 2.40 A resolution structure of drug-free MepR reveals the most open MarR family protein conformation to date, which will require a huge conformational change to bind cognate DNA. DNA-binding data show that MepR uses a dual regulatory binding mode as the repressor binds the mepA operator as a dimer of dimers, but binds the mepR operator as a single dimer. Alignment of the six half sites reveals the consensus MepR binding site, 5'-GTTAGAT-3'. 'Drug' binding studies show that MepR binds to ethidium and DAPI with comparable affinities (K(d) = 2.6 and 4.5 microM, respectively), but with significantly lower affinity to the larger rhodamine 6G (K(d) = 62.6 microM). Mapping clinically relevant or in vitro selected MepR mutants onto the MepR structure suggests that their defective repressor phenotypes are due to structural and allosteric defects. PMID- 19129226 TI - Insights into the pre-initiation events of bacteriophage phi 6 RNA-dependent RNA polymerase: towards the assembly of a productive binary complex. AB - The RNA-dependent RNA polymerase (RdRP) of double-stranded RNA (dsRNA) viruses performs both RNA replication and transcription. In order to initiate RNA polymerization, viral RdRPs must be able to interact with the incoming 3' terminus of the template and position it, so that a productive binary complex is formed. Structural studies have revealed that RdRPs of dsRNA viruses that lack helicases have electrostatically charged areas on the polymerase surface, which might facilitate such interactions. In this study, structure-based mutagenesis, enzymatic assays and molecular mapping of bacteriophage phi 6 RdRP and its RNA were used to elucidate the roles of the negatively charged plough area on the polymerase surface, of the rim of the template tunnel and of the template specificity pocket that is key in the formation of the productive RNA-polymerase binary complex. The positively charged rim of the template tunnel has a significant role in the engagement of highly structured ssRNA molecules, whereas specific interactions further down in the template tunnel promote ssRNA entry to the catalytic site. Hence, we show that by aiding the formation of a stable binary complex with optimized RNA templates, the overall polymerization activity of the phi 6 RdRP can be greatly enhanced. PMID- 19129227 TI - Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction. AB - Here we have developed a sensitive DNA amplified detection method based on isothermal strand-displacement polymerization reaction. This method takes advantage of both the hybridization property of DNA and the strand-displacement property of polymerase. Importantly, we demonstrate that our method produces a circular polymerization reaction activated by the target, which essentially allows it to self-detect. Functionally, this DNA system consists of a hairpin fluorescence probe, a short primer and polymerase. Upon recognition and hybridization with the target ssDNA, the stem of the hairpin probe is opened, after which the opened probe anneals with the primer and triggers the polymerization reaction. During this process of the polymerization reaction, a complementary DNA is synthesized and the hybridized target is displaced. Finally, the displaced target recognizes and hybridizes with another probe, triggering the next round of polymerization reaction, reaching a target detection limit of 6.4 x 10(-15) M. PMID- 19129228 TI - Telomerase activity is associated with an increase in DNA methylation at the proximal subtelomere and a reduction in telomeric transcription. AB - Tumours and immortalized cells avoid telomere attrition by using either the ribonucleoprotein enzyme telomerase or a recombination-based alternative lengthening of telomeres (ALT) mechanism. Available evidence from mice suggests that the epigenetic state of the telomere may influence the mechanism of telomere maintenance, but this has not been directly tested in human cancer. Here we investigated cytosine methylation directly adjacent to the telomere as a marker of the telomere's epigenetic state in a panel of human cell lines. We find that while ALT cells show highly heterogeneous patterns of subtelomeric methylation, subtelomeric regions in telomerase-positive cells invariably show denser methylation than normal cells, being almost completely methylated. When compared to matched normal and ALT cells, telomerase-positive cells also exhibit reduced levels of the telomeric repeat-containing-RNA (TERRA), whose transcription originates in the subtelomere. Our results are consistent with the notion that TERRA may inhibit telomerase: the heavy cytosine methylation we observe in telomerase-positive cells may reflect selection for TERRA silencing in order to facilitate telomerase activity at the telomere. These data suggest that the epigenetic differences between telomerase-positive and ALT cells may underlie the mechanism of telomere maintenance in human tumorigenesis and highlight the broad reaching consequences of epigenetic dysregulation in cancer. PMID- 19129229 TI - Telomere length measurement by a novel monochrome multiplex quantitative PCR method. AB - The current quantitative polymerase chain reaction (QPCR) assay of telomere length measures telomere (T) signals in experimental DNA samples in one set of reaction wells, and single copy gene (S) signals in separate wells, in comparison to a reference DNA, to yield relative T/S ratios that are proportional to average telomere length. Multiplexing this assay is desirable, because variation in the amount of DNA pipetted would no longer contribute to variation in T/S, since T and S would be collected within each reaction, from the same input DNA. Multiplexing also increases throughput and lowers costs, since half as many reactions are needed. Here, we present the first multiplexed QPCR method for telomere length measurement. Remarkably, a single fluorescent DNA-intercalating dye is sufficient in this system, because T signals can be collected in early cycles, before S signals rise above baseline, and S signals can be collected at a temperature that fully melts the telomere product, sending its signal to baseline. The correlation of T/S ratios with Terminal Restriction Fragment (TRF) lengths measured by Southern blot was stronger with this monochrome multiplex QPCR method (R(2) = 0.844) than with our original singleplex method (R(2) = 0.677). Multiplex T/S results from independent runs on different days were highly reproducible (R(2) = 0.91). PMID- 19129230 TI - Molecular characterization of Mybbp1a as a co-repressor on the Period2 promoter. AB - The circadian clock comprises transcriptional feedback loops of clock genes. Cryptochromes are essential components of the negative feedback loop in mammals as they inhibit CLOCK-BMAL1-mediated transcription. We purified mouse CRY1 (mCRY1) protein complexes from Sarcoma 180 cells to determine their roles in circadian gene expression and discovered that Myb-binding protein 1a (Mybbp1a) interacts with mCRY1. Mybbp1a regulates various transcription factors, but its role in circadian gene expression is unknown. We found that Mybbp1a functions as a co-repressor of Per2 expression and repressed Per2 promoter activity in reporter assays. Chromatin immunoprecipitation (ChIP) assays revealed endogenous Mybbp1a binding to the Per2 promoter that temporally matched that of mCRY1. Furthermore, Mybbp1a binding to the Per2 promoter correlated with the start of the down-regulation of Per2 expression and with the dimethylation of histone H3 Lys9, to which it could also bind. These findings suggest that Mybbp1a and mCRY1 can form complexes on the Per2 promoter that function as negative regulators of Per2 expression. PMID- 19129231 TI - The N-terminal PIN domain of the exosome subunit Rrp44 harbors endonuclease activity and tethers Rrp44 to the yeast core exosome. AB - Nuclear and cytoplasmic forms of the yeast exosome share 10 components, of which only Rrp44/Dis3 is believed to possess 3' exonuclease activity. We report that expression only of Rrp44 lacking 3'-exonuclease activity (Rrp44-exo) supports growth in S288c-related strains (BY4741). In BY4741, rrp44-exo was synthetic lethal with loss of the cytoplasmic 5'-exonuclease Xrn1, indicating block of mRNA turnover, but not with loss of the nuclear 3'-exonuclease Rrp6. The RNA processing phenotype of rrp44-exo was milder than that seen on Rrp44 depletion, indicating that Rrp44-exo retains important functions. Recombinant Rrp44 was shown to possess manganese-dependent endonuclease activity in vitro that was abolished by four point mutations in the putative metal binding residues of its N terminal PIN domain. Rrp44 lacking both exonuclease and endonuclease activity failed to support growth in strains depleted of endogenous Rrp44. Strains expressing Rrp44-exo and Rrp44-endo-exo exhibited different RNA processing patterns in vivo suggesting Rrp44-dependent endonucleolytic cleavages in the 5' ETS and ITS2 regions of the pre-rRNA. Finally, the N-terminal PIN domain was shown to be necessary and sufficient for association with the core exosome, indicating its dual function as a nuclease and structural element. PMID- 19129232 TI - Positioning of subdomain IIId and apical loop of domain II of the hepatitis C IRES on the human 40S ribosome. AB - The 5'-untranslated region of the hepatitis C virus (HCV) RNA contains a highly structured motif called IRES (Internal Ribosome Entry Site) responsible for the cap-independent initiation of the viral RNA translation. At first, the IRES binds to the 40S subunit without any initiation factors so that the initiation AUG codon falls into the P site. Here using an original site-directed cross-linking strategy, we identified 40S subunit components neighboring subdomain IIId, which is critical for HCV IRES binding to the subunit, and apical loop of domain II, which was suggested to contact the 40S subunit from data on cryo-electron microscopy of ribosomal complexes containing the HCV IRES. HCV IRES derivatives that bear a photoactivatable group at nucleotide A275 or at G263 in subdomain IIId cross-link to ribosomal proteins S3a, S14 and S16, and HCV IRES derivatized at the C83 in the apex of domain II cross-link to proteins S14 and S16. PMID- 19129233 TI - Mechanism of DNA flexibility enhancement by HMGB proteins. AB - The mechanism by which sequence non-specific DNA-binding proteins enhance DNA flexibility is studied by examining complexes of double-stranded DNA with the high mobility group type B proteins HMGB2 (Box A) and HMGB1 (Box A+B) using atomic force microscopy. DNA end-to-end distances and local DNA bend angle distributions are analyzed for protein complexes deposited on a mica surface. For HMGB2 (Box A) binding we find a mean induced DNA bend angle of 78 degrees, with a standard error of 1.3 degrees and a SD of 23 degrees, while HMGB1 (Box A+B) binding gives a mean bend angle of 67 degrees, with a standard error of 1.3 degrees and a SD of 21 degrees. These results are consistent with analysis of the observed global persistence length changes derived from end-to-end distance measurements, and with results of DNA-stretching experiments. The moderately broad distributions of bend angles induced by both proteins are inconsistent with either a static kink model, or a purely flexible hinge model for DNA distortion by protein binding. Therefore, the mechanism by which HMGB proteins enhance the flexibility of DNA must differ from that of the Escherichia coli HU protein, which in previous studies showed a flat angle distribution consistent with a flexible hinge model. PMID- 19129234 TI - Cisplatin induces loop structures and condensation of single DNA molecules. AB - Structural properties of single lambda DNA treated with anti-cancer drug cisplatin were studied with magnetic tweezers and AFM. Under the effect of low concentration cisplatin, the DNA became more flexible, with the persistence length decreased significantly from approximately 52 to 15 nm. At a high drug concentration, a DNA condensation phenomenon was observed. Based on experimental results from both single-molecule and AFM studies, we propose a model to explain this kind of DNA condensation by cisplatin: first, di-adducts induce local distortions of DNA. Next, micro-loops of approximately 20 nm appear through distant crosslinks. Then, large aggregates are formed through further crosslinks. Finally, DNA is condensed into a compact globule. Experiments with Pt(dach)Cl(2) indicate that oxaliplatin may modify the DNA structures in the same way as cisplatin. The observed loop structure formation of DNA may be an important feature of the effect of platinum anti-cancer drugs that are analogous to cisplatin in structure. PMID- 19129235 TI - A role for monoubiquitinated FANCD2 at telomeres in ALT cells. AB - Both Fanconi anemia (FA) and telomere dysfunction are associated with chromosome instability and an increased risk of cancer. Because of these similarities, we have investigated whether there is a relationship between the FA protein, FANCD2 and telomeres. We find that FANCD2 nuclear foci colocalize with telomeres and PML bodies in immortalized telomerase-negative cells. These cells maintain telomeres by alternative lengthening of telomeres (ALT). In contrast, FANCD2 does not colocalize with telomeres or PML bodies in cells which express telomerase. Using a siRNA approach we find that FANCA and FANCL, which are components of the FA nuclear core complex, regulate FANCD2 monoubiquitination and the telomeric localization of FANCD2 in ALT cells. Transient depletion of FANCD2, or FANCA, results in a dramatic loss of detectable telomeres in ALT cells but not in telomerase-expressing cells. Furthermore, telomere loss following depletion of these proteins in ALT cells is associated with decreased homologous recombination between telomeres (T-SCE). Thus, the FA pathway has a novel function in ALT telomere maintenance related to DNA repair. ALT telomere maintenance is therefore one mechanism by which monoubiquitinated FANCD2 may promote genetic stability. PMID- 19129236 TI - Gene expression modulation is associated with gene amplification, supernumerary chromosomes and chromosome loss in antimony-resistant Leishmania infantum. AB - Antimonials remain the first line drug against the protozoan parasite Leishmania but their efficacy is threatened by resistance. We carried out a RNA expression profiling analysis comparing an antimony-sensitive and -resistant (Sb2000.1) strain of Leishmania infantum using whole-genome 70-mer oligonucleotide microarrays. Several genes were differentially expressed between the two strains, several of which were found to be physically linked in the genome. MRPA, an ATP binding cassette (ABC) gene known to be involved in antimony resistance, was overexpressed in the antimony-resistant mutant along with three other tandemly linked genes on chromosome 23. This four gene locus was flanked by 1.4 kb repeated sequences from which an extrachromosomal circular amplicon was generated in the resistant cells. Interestingly, gene expression modulation of entire chromosomes occurred in the antimony-resistant mutant. Southern blots analyses and comparative genomic hybridizations revealed that this was either due to the presence of supernumerary chromosomes or to the loss of one chromosome. Leishmania parasites with haploid chromosomes were viable. Changes in copy number for some of these chromosomes were confirmed in another antimony-resistant strain. Selection of a partial revertant line correlated antimomy resistance levels and the copy number of aneuploid chromosomes, suggesting a putative link between aneuploidy and drug resistance in Leishmania. PMID- 19129237 TI - RNA structure prediction from evolutionary patterns of nucleotide composition. AB - Structural elements in RNA molecules have a distinct nucleotide composition, which changes gradually over evolutionary time. We discovered certain features of these compositional patterns that are shared between all RNA families. Based on this information, we developed a structure prediction method that evaluates candidate structures for a set of homologous RNAs on their ability to reproduce the patterns exhibited by biological structures. The method is named SPuNC for 'Structure Prediction using Nucleotide Composition'. In a performance test on a diverse set of RNA families we demonstrate that the SPuNC algorithm succeeds in selecting the most realistic structures in an ensemble. The average accuracy of top-scoring structures is significantly higher than the average accuracy of all ensemble members (improvements of more than 20% observed). In addition, a consensus structure that includes the most reliable base pairs gleaned from a set of top-scoring structures is generally more accurate than a consensus derived from the full structural ensemble. Our method achieves better accuracy than existing methods on several RNA families, including novel riboswitches and ribozymes. The results clearly show that nucleotide composition can be used to reveal the quality of RNA structures and thus the presented technique should be added to the set of prediction tools. PMID- 19129239 TI - Occupational eye injuries: a continuing problem. AB - BACKGROUND: Ocular trauma is a worldwide cause of visual morbidity, a significant proportion of which occurs in the workplace. This is largely preventable with the use of protective eyewear and strict compliance. AIM: To analyse the type of occupational eye injuries that occur and to document the use of eye protection in patients presenting to a UK district general hospital. METHODS: A pilot retrospective case note analysis of all ocular injuries seen in one calendar month was performed. A prospective survey of consecutive occupational ocular injuries presenting to this district general accident and emergency (A&E) department over a 2-month period was then carried out. Demographics, aetiology, eye protection usage and clinical details were recorded and analysed. RESULTS: Of all eye patients attending this A&E, 31% (87/283) were due to occupational eye injuries. Of 55 prospectively reviewed patients with occupational eye injuries, the majority had minor injuries. Of the cases where eye protection was recorded, 56% (18/32) were not wearing any protection and 44% (14/32) wore eye protection at the time of injury. CONCLUSIONS: Occupational eye injuries are a commonly seen ocular complaint in the A&E department. Provision of appropriate eyewear protection and worker education is required to minimize the incidence of ocular injury in the workplace. PMID- 19129238 TI - Consumption of SC45647 and sucralose by rats selectively bred for high and low saccharin intake. AB - Mammals' affinity for sweet tastes exists alongside dramatic variation among species and individuals in responses to sweeteners. The present paper focused on consumption by Occidental High- (HiS) and Low-Saccharin (LoS)-consuming rats in 23-h 2-bottle tests of 2 sweeteners for which few data from rats are available: SC45647 and sucralose. Every HiS and LoS rat preferred SC45647 to water at every concentration, with HiS rats consuming it more avidly. Most HiS rats preferred sucralose to water at one or more concentrations; some HiS rats and most LoS rats avoided sucralose at every concentration. However, both HiS and LoS rats preferred a sucralose-maltodextrin mixture (Splenda) to water; thus, Splenda's "bulking" ingredient maltodextrin transforms highly variable responses to sucralose into a relatively homogeneous preference for the product. Implications for the study of variation in sweet taste are discussed. PMID- 19129240 TI - Attitudes to job turnover among Finnish anaesthetists. AB - BACKGROUND: Structural changes have led to higher workload and more frequent conflicts among hospital staff, which in turn has been shown to be associated with increased employee turnover. AIMS: To study the willingness of anaesthetists to change their employment and factors associated with it. Work-related, individual and family-related factors were investigated as potential influences on such willingness. METHOD: A postal questionnaire was sent to all working Finnish anaesthetists (N = 550). RESULTS: The response rate was 60%; 175 (53% of responders) were men. Of the respondents, 31% were willing to consider changing to another physician's job and 43% to a profession other than medicine. The most important correlates for these views were conflicts with superiors (odds ratio 6.1; 95% confidence interval 2.1-17.7) and co-workers (4.2; 1.4-12.2), low job control (2.6; 1.4-4.9), a sense of organizational injustice (2.4; 1.3-4.6), stress (6.5; 2.6-16.3) and job dissatisfaction (4.6; 2.4-8.8). CONCLUSIONS: The establishment of respect, trust and genuine dialogue between co-workers and superiors is needed to minimize the risk of loss of members of this occupational group. PMID- 19129241 TI - Autosomal dominant Alport syndrome: molecular analysis of the COL4A4 gene and clinical outcome. AB - BACKGROUND: Alport syndrome is a clinically and genetically heterogeneous nephropathy characterized by glomerular basement membrane lesions often associated with hearing loss and ocular anomalies. While the X-linked and the autosomal recessive forms are well known, the autosomal dominant form is not well acknowledged. METHODS: We have clinically investigated 38 patients with a diagnosis of autosomal dominant Alport syndrome belonging to eight different families. The analysis of the COL4A4 gene was performed by denaturing high performance liquid chromatography and automated DNA sequencing. RESULTS: In our cohort of patients, only 24.3% (9/37) reached end-stage renal disease, at the mean age of 51.2 years. Four patients had hearing loss (13.3%) and none ocular changes. Molecular analysis revealed eight novel private COL4A4 gene mutations: three frameshift, three missense and two splice-site mutations. CONCLUSIONS: These data indicate autosomal dominant Alport syndrome as a disease with a low risk of ocular and hearing anomalies but with a significant risk to develop renal failure although at an older age than the X-linked form. We were unable to demonstrate a genotype-phenotype correlation. Altogether, these data make difficult the differential diagnosis with the benign familial haematuria due to heterozygous mutations of COL4A4 and COL4A3, especially in young patients, and with the X-linked form of Alport syndrome in families where only females are affected. A correct diagnosis and prognosis is based on a comprehensive clinical investigation in as many family members as possible associated with a broadly formal genetic analysis of the pedigree. PMID- 19129242 TI - LPA1 receptors mediate stimulation, whereas LPA2 receptors mediate inhibition, of migration of pancreatic cancer cells in response to lysophosphatidic acid and malignant ascites. AB - Malignant ascites from pancreatic cancer patients has been reported to stimulate migration of pancreatic cancer cells through lysophosphatidic acid (LPA) and LPA(1) receptors. Indeed, ascites- and LPA-induced migration was inhibited by Ki16425, an LPA(1) and LPA(3) antagonist, in Panc-1 cells. Unexpectedly, however, in the presence of Ki16425, ascites and LPA inhibited cell migration in response to epidermal growth factor (EGF). The inhibitory migratory response to ascites and LPA was also observed in the cells treated with pertussis toxin (PTX), a G(i) protein inhibitor, and attenuated by a small interfering RNA (siRNA) specific to the LPA(2) receptor. The inhibitory LPA action was reversed by the regulators of G-protein signaling domain of p115RhoGEF, dominant-negative RhoA or C3 toxin. Indeed, LPA activated RhoA, which was attenuated by the siRNA against the LPA(2) receptor. Moreover, LP-105, an LPA(2) agonist, also inhibited EGF-induced migration in the PTX-treated cells. A similar inhibitory migration response through LPA(2) receptors was also observed in YAPC-PD, BxPC-3, CFPAC-1 and PK-1 pancreatic cancer cell lines. LPA also inhibited the invasion of Panc-1 cells in the PTX-treated cells in the in vitro Matrigel invasion assay. We conclude that LPA(2) receptors are coupled to the G(12/13) protein/Rho-signaling pathway, leading to the inhibition of EGF-induced migration and invasion of pancreatic cancer cells. PMID- 19129243 TI - Heparin-induced thrombocytopenia with thrombosis syndrome managed with plasmapheresis. AB - Heparin-induced thrombocytopenia with thrombosis syndrome is an antibody-mediated disorder that has a high mortality in cardiac surgical patients in spite of early diagnosis and management with direct thrombin inhibitors. Plasmapheresis, an extracorporeal technique that has been designed for the removal of large molecular weight substances from the plasma, can remove the offending antibodies from these desperately ill patients. We describe a case of a postoperative cardiac surgery patient with heparin-induced thrombocytopenia with thrombosis syndrome and multi-system failure who was dependent upon a left ventricular assist device. He was treated successfully with plasmapheresis with recovery of his platelet count from 25,000/microl to over 200,000/microl, along with multi organ recovery. This patient survived because of plasmapheresis. Removing the antibodies to the heparin-platelet factor four complex with plasmapheresis is an effective strategy to treat these patients. We believe that the use of plasmapheresis as a bail-out procedure in these often desperately ill post operative cardiac surgical patients who have heparin-induced thrombocytopenia with thrombosis syndrome could be lifesaving. PMID- 19129244 TI - Adverse influence of female gender on outcomes after coronary bypass surgery: a propensity matched analysis. AB - Female patients have been shown to have a different risk profile and increased adverse events after coronary bypass grafting (CAB). The objective was to create a risk profile based on gender and look at outcomes in propensity matched groups with similar factors differing by gender. From May 2001 to December 2006, 976 patients underwent isolated CAB. Univariable analysis created a risk profile for female and male patients. Multivariable logistic regression was used to develop independent predictors of mortality. Propensity matching for the most predictive variables of adverse was used to create matched sets of 269 male and female patients to analyze outcomes independent of these variables. Of the 976 patients 31.7% were female. Of the 19 preoperative risk factors analyzed, diabetes, hypertension, older age, higher body mass index (BMI), African-American race were more predictive of female gender. Males had more smokers and acute myocardial infarction within seven days before surgery. Independent predictors of mortality were female gender (P=0.01), diabetes (P=0.02), increased age (P=0.02), acute MI <7 days (P=0.003). Propensity matching (for smoking, diabetes, hypertension, MI <7 days, age, BMI and race) still showed increased in-hospital complications and mortality for female patients. Female patients undergoing CAB have a different risk profile and have a higher incidence of adverse outcomes including death, which are not mitigated by careful matching with male patients. PMID- 19129245 TI - Highly fucosylated N-glycan ligands for mannan-binding protein expressed specifically on CD26 (DPPVI) isolated from a human colorectal carcinoma cell line, SW1116. AB - The serum mannan-binding protein (MBP) is a host defense C-type lectin specific for mannose, N-acetylglucosamine, and fucose residues, and exhibits growth inhibitory activity toward human colorectal carcinoma cells. The MBP-ligand oligosaccharides (MLO) isolated from a human colorectal carcinoma cell line, SW1116, are large, multiantennary N-glycans with highly fucosylated polylactosamine-type structures having Le(b)-Le(a) or tandem repeats of the Le(a) structure at their nonreducing ends. In this study, we isolated the major MBP ligand glycoproteins from SW1116 cell lysates with an MBP column and identified them as CD26/dipeptidyl peptidase IV (DPPIV) (110 kDa) and CD98 heavy chain (CD98hc)/4F2hc (82 kDa). Glycosidase digestion revealed that CD26 contained such complex-type N-glycans that appear to mediate the MBP binding. MALDI-MS of the N glycans released from CD26 by PNGase F demonstrated conclusively that CD26 is the major MLO-carrying protein. More interestingly, a comparison of the N-glycans released from the MBP-binding and non-MBP-binding glycopeptides suggested that complex-type N-glycans carrying a minimum of 4 Le(a)/Le(b) epitopes arranged either as multimeric tandem repeats or terminal epitopes on multiantennary structures are critically important for the high affinity binding to MBP. Analysis of the N-glycan attachment sites demonstrated that the high affinity MLO was expressed preferentially at some N-glycosylation sites, but this site preference was not so stringent. Finally, hypothetical 3D models of tandem repeats of the Le(a) epitope and the MBP-Lewis oligosaccharide complex were presented. PMID- 19129246 TI - Hetero-oligomeric interactions between early glycosyltransferases of the dolichol cycle. AB - N-Linked glycosylation begins with the formation of a dolichol-linked oligosaccharide in the endoplasmic reticulum (ER). The first two steps of this pathway lead to the formation of GlcNAc(2)-PP-dolichol, whose synthesis is sequentially catalyzed by the Alg7p GlcNAc phosphotransferase and by the dimeric Alg13p/Alg14p UDP-GlcNAc transferase on the cytosolic face of the endoplasmic reticulum. Here, we show that the Alg7p, Alg13p, and Alg14p glycosyltransferases form a functional multienzyme complex. Coimmunoprecipitation and gel filtration assays demonstrate that the Alg7p/Alg13p/Alg14p complex is a hexamer with a native molecular weight of approximately 200 kDa and an Alg7p:Alg13:Alg14p stoichiometry of 1:1:1. These results highlight and extend the striking parallels that exist between these eukaryotic UDP-GlcNAc transferases and their bacterial MraY and MurG homologs that catalyze the first two steps of the lipid-linked peptidoglycan precursor. In addition to their preferred substrate and lipid acceptors, these enzymes are similar in their structure, chemistry, temporal, and spatial organization. These similarities point to an evolutionary link between the early steps of N-linked glycosylation and those of peptidoglycan synthesis. PMID- 19129247 TI - Development of valuable yeast strains using a novel mutagenesis technique for the effective production of therapeutic glycoproteins. AB - Yeast cells producing mammalian-type N-linked oligosaccharide show severe growth defects and the decreased protein productivity because of the disruption of yeast specific glycosyltransferases. This decreased protein productivity in engineered yeast strains is an obstacle to the development of efficient glycoprotein production in yeast. For economic and effective synthesis of such therapeutic glycoproteins in yeast, the development of appropriate strains is highly desirable. We applied a novel mutagenesis technique that utilized the proofreading-deficient DNA polymerase delta variant encoded by the pol3-01 gene of Saccharomyces cerevisiae or the cdc6-1 gene of Schizosaccharomyces pombe to the engineered S. cerevisiae TIY20 strain and S. pombe KT97 strain, respectively. TIY20, which is deficient in the outer chain of mannan due to the disruption of three genes (och1Delta, mnn1 Delta, mnn4 Delta), and KT97, which is an och1 disruptant, are impractical as hosts for the production of therapeutic glycoproteins since they show a temperature-sensitive (ts) phenotype, a growth defect phenotype, and decreased protein productivity. We successfully isolated YAB mutants that alleviated the growth defect of the TIY20 strain. Surprisingly, these mutants generally secreted foreign proteins better than the wild-type strain. Furthermore, we successfully isolated YPAB mutants that alleviated the growth defect of the KT97 strain, too. The development of these new mutants by the combination of genetic engineering of yeast and this mutagenesis technique are major breakthroughs for the production of therapeutic glycoproteins in engineered yeast cells. PMID- 19129248 TI - Classical cardiovascular risk factors and all-cause mortality in rural Cameroon. AB - BACKGROUND: High levels of cardiovascular risk factors have been reported in rural Africa. How these translate into major outcomes remain unknown. AIM: To assess the association between selected risk factors and all-cause mortality in rural Cameroon. DESIGN: Nine-year's prospective cohort study. METHODS: The 9 year's vital status was ascertained for 350 participants screened for cardiovascular risk factors in Bafut, Cameroon in 1998. Cox models were used to compute the hazard ratio (HR) and 95% confidence interval (CI) for selected risk factors. RESULTS: Vital status was available for all participants, except 22 (6.3%) who were excluded from analyses. At baseline, compared with women, men had significantly higher waist-to-hip ratio, were more likely to be ex- or current smokers and alcohol consumers (all P < or = 0.008). The total duration of follow up was 2771 person-years. This duration was longer for women (P = 0.04). During follow-up 47 deaths were recorded, 31 (66%) in men (P = 0.023). In multivariate Cox analyses, age, male gender, current smoking, systolic blood pressure and fasting capillary glucose were significant predictors of total mortality during follow-up. CONCLUSION: Gender, smoking, fasting capillary glucose, blood pressures and age are potential determinants of overall death in rural Cameroon. More elaborated cohort studies are needed to refine these conclusions and monitor the progression of these populations through epidemiological transition stages. PMID- 19129249 TI - Renal function is associated with arterial stiffness and predicts outcome in patients with coronary artery disease. AB - BACKGROUND: Arterial stiffness (AS) is associated, and a predictor of, outcome in patients with cardiovascular and renal disease. AIM: In this study, we estimated glomerular filtration rate (eGFR) and measured indices of AS in patients with suspected coronary artery disease (CAD), and assessed their predictive value on outcome. DESIGN: Prospective cohort study. METHODS: AS was measured using pulse wave velocity (PWV) and pulse wave analysis in patients with no known renal disease who had recently undergone coronary angiography. Renal function was assessed using serum creatinine concentration [creat](sr) and eGFR (Cockcroft & Gault, C&G). The primary endpoint was a combination of hospitalization due to cardiovascular disease and all-cause mortality. RESULTS: Two hundred eighty-four subjects (210 men, 74 women, mean age 62 years) were followed-up for a mean of 1.5 years. PWV was negatively associated with eGFR (r(2) = 0.09, P < 0.001), even in patients with an eGFR > or =60 ml/min/m(2) (r(2) = 0.04, P < 0.01). PWV was determined by age, heart rate, systolic blood pressure, body mass index and [creat](sr) (r(2) = 0.38, P < 0.001). A lower eGFR (P < 0.01), PWV above the median (P < 0.05) and degree of CAD (P < 0.001) predicted a shorter time to the primary endpoint. eGFR and degree of CAD remained independent determinants of outcomes (P < 0.01), even in patients with normal renal function (P < 0.01). CONCLUSION: This study suggests that even minor reductions in eGFR, within the normal range, are an additional independent risk marker in patients with CAD. PMID- 19129250 TI - Alcohol--it's more than the liver. PMID- 19129251 TI - Aicardi-Goutieres syndrome. AB - INTRODUCTION: Aicardi-Goutieres syndrome (AGS) is an autosomal recessive encephalopathy characterized by acquired microcephaly, cerebral calcifications, leukodystrophy, cerebral atrophy and cerebrospinal fluid findings of chronic lymphocytosis and raised interferon-alpha (INF-alpha). The main extraneurological symptoms are chilblain-like skin lesions, usually on the fingers, toes and ears. SOURCES OF DATA: This review is based on a search of the published literature on AGS from 1984 onwards (particularly the most recent papers) and on knowledge and experience gained through the authors' work with the International Aicardi Goutieres Syndrome Association (IAGSA). AREAS OF AGREEMENT: It is accepted that AGS can be mistaken for a congenital infection and that the diagnostic significance of its cardinal signs (raised INF-alpha levels, basal ganglia calcifications) is different in different stages of the disease. Currently, we know of four genes that, if mutated, can give rise to AGS, but at least one other gene is believed to exist. These genes are involved in the DNA damage response, a defect of which could provoke an inappropriate innate immune response, triggering increased secretion of INF-alpha, ultimately responsible for the main features of the disease. AREAS OF CONTROVERSY: The natural history of AGS has not yet been definitively described given the lack of extensive, long-term neuroradiological follow-up studies. Furthermore, it is not yet clearly understood how the innate immune system is activated, what triggers the onset of the disease or why it tends to 'burn out' after several months. Immunosuppressive therapy in the active stage of the disease does not seem to produce any real change in the clinical course, but more data are needed. GROWING POINTS AND AREAS TIMELY FOR DEVELOPING RESEARCH: Current studies aim to clarify the molecular mechanisms underlying the pathogenesis of AGS and to establish the exact pathway by which retained nucleic acids activate the immune system. This knowledge could allow the development of therapeutic strategies. PMID- 19129252 TI - Chronic blockade of 20-HETE synthesis reduces polycystic kidney disease in an orthologous rat model of ARPKD. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE) has been implicated as a potential mediator in epithelial cell proliferation and cyst formation in polycystic kidney disease (PKD). In the present study, we studied the effects of chronic blockade of 20-HETE synthesis in an orthologous rodent model of autosomal recessive polycystic kidney disease (ARPKD), the PCK rat. RT-PCR analysis indicated that the expression of CYP4A1, CYP4A2, CYP4A3, and CYP4A8 mRNA was increased two- to fourfold in cystic PCK compared with noncystic Sprague-Dawley rat kidneys. Daily administration of a 20-HETE synthesis inhibitor, HET-0016 (10 mg x kg(-1) x day( 1) ip) for 4-7 wk significantly reduced kidney size by 24% from 4.95 +/- 0.19 g in vehicle-treated PCK rats to 3.76 +/- 0.15 g (n = 4). Collecting tubule morphometric cystic indices were reduced in HET-0016-treated PCK rats (2.1 +/- 0.2; n = 4) compared with vehicle-treated PCK rats (4.4 +/- 0.1; n = 4). The cellular mechanism by which 20-HETE may play a role in cyst formation has not been well characterized, but there was a significantly lower (P < 0.05) level of intracellular cAMP and decreased phosphorylation (activation) of ERK1/2 protein in PCK rat kidneys (n = 3) treated with HET-0016 . These studies indicate a potential role of 20-HETE in cyst formation in the orthologous rodent PCK model of ARPKD. PMID- 19129253 TI - Rho-kinase inhibition reduces pressure-mediated autoregulatory adjustments in afferent arteriolar diameter. AB - Preglomerular resistance is regulated by calcium influx- and mobilization dependent mechanisms; however, the role of Rho-kinase in calcium sensitization in the intact kidney has not been carefully examined. Experiments were performed to test the hypothesis that Rho-kinase inhibition blunts pressure-mediated afferent arteriolar autoregulatory behavior and vasoconstrictor responses evoked by angiotensin II and P2X1 receptor activation. Rat kidneys were studied in vitro using the blood-perfused juxtamedullary nephron technique. Autoregulatory behavior was assessed before and during Rho-kinase inhibition with Y-27632 (1.0 microM; n = 5). Control diameter averaged 14.3 +/- 0.8 microm and increased to 18.1 +/- 0.9 microm (P < 0.05) during Y-27632 treatment. In the continued presence of Y-27632, reducing perfusion pressure to 65 mmHg slightly increased diameter to 18.7 +/- 1.0 microm. Subsequent pressure increases to 130 and 160 mmHg yielded afferent arteriolar diameters of 17.5 +/- 0.8 and 16.6 +/- 0.6 microm (P < 0.05). This 11% decline in diameter is significantly smaller than the 40% decrease obtained in untreated kidneys. The inhibitory effects of Y-27632 on autoregulatory behavior were concentration dependent. Angiotensin II responses were blunted by Y-27632. Angiotensin II (1.0 nM) reduced afferent diameter by 17 +/- 1% in untreated arterioles and by 6 +/- 2% during exposure to Y-27632. The P2X1 receptor agonist, alpha, beta-methylene ATP, reduced afferent arteriolar diameter by 8 +/- 1% but this response was eliminated during exposure to Y-27632. Western blot analysis confirms expression of the Rho-kinase signaling pathway. Thus, Rho-kinase may be important in pressure-mediated autoregulatory adjustments in preglomerular resistance and responsiveness to angiotensin II and autoregulatory P2X1 receptor agonists. PMID- 19129254 TI - Basolateral expression of the ammonia transporter family member Rh C glycoprotein in the mouse kidney. AB - Ammonia metabolism and transport are critical for acid-base homeostasis. The ammonia transporter family member Rh C glycoprotein (Rhcg) is expressed in distal renal tubular segments, and its expression is regulated in parallel with renal ammonia metabolism. However, there are inconsistencies in its reported subcellular distribution, with both apical and basolateral Rhcg reported in rat and human kidney and only apical expression in mouse kidney. Because the membrane location of Rhcg is critical for understanding its physiological role, we reassessed mouse Rhcg localization using refined immunolocalization methods. Two antibodies directed against different Rhcg-specific epitopes identified both apical and basolateral Rhcg immunolabel in mouse kidney. Immunogold electron microscopy both confirmed basolateral plasma membrane Rhcg expression and showed that apical immunolabel represented expression in both the apical plasma membrane and in subapical cytoplasmic vesicles. Immunoblots and Northern blots identified similar bands in Balb/c and C57BL/6 kidneys, suggesting basolateral Rhcg may result from alternative trafficking. Basolateral Rhcg intensity was strain dependent, with less basolateral Rhcg expression in the Balb/c mouse compared with the C57BL/6 mouse. In mice with collecting duct-specific Rhcg gene deletion, generated using Cre-loxP techniques, neither apical nor basolateral Rhcg immunolabel was identified in the collecting duct, confirming that basolateral Rhcg was the product of the same gene product as apical Rhcg. Although basolateral Rhcg expression differed between C57BL/6 and Balb/c mice, Rh B glycoprotein, which is exclusively basolateral, was expressed at similar levels in the two strains. We conclude that Rhcg is present in both the apical and basolateral plasma membrane in the mouse kidney, where it is likely to contribute to renal ammonia metabolism. PMID- 19129255 TI - Macrophages are not the source of injurious interleukin-18 in ischemic acute kidney injury in mice. AB - We previously reported in ischemic acute kidney injury (AKI) in mice that caspase 1-mediated production of interleukin-18 (IL-18) is pathogenic and that macrophage depletion by liposome-encapsulated clodronate (LEC) is protective. Therefore, our aim was to determine whether macrophages are a source of IL-18 in ischemic AKI in mice. On immunofluorescence staining of the outer stripe of outer medulla, the number of macrophages double stained for CD11b and IL-18 was significantly increased in AKI and significantly decreased by LEC. Adoptive transfer of RAW 264.7 cells, a mouse macrophage line that constitutively expresses IL-18 mRNA, reversed the functional protection against AKI in both LEC-treated wild-type and caspase-1 -/- mice. To test whether IL-18 in macrophages is necessary to cause AKI, we adoptively transferred macrophages in which IL-18 was inhibited. Peritoneal macrophages isolated from wild-type mice, IL-18 binding protein transgenic (IL-18 BP Tg) mice, and IL-18 -/- mice were used. IL-18 BP Tg mice overexpress human IL-18 BP and exhibit decreased biological activity of IL-18. Adoptive transfer of peritoneal macrophages from wild-type as well as IL-18 BP Tg and IL-18 -/- mice reversed the functional protection against AKI in LEC-treated mice. In summary, adoptive transfer of RAW cells, that constitutively express IL 18, reverses the functional protection in macrophage-depleted wild-type and caspase-1 -/- mice with AKI. However, adoptive transfer of peritoneal macrophages in which IL-18 function was inhibited also reverses the functional protection in macrophage-depleted mice. In conclusion, IL-18 from adoptive transfer of macrophages is not sufficient to cause ischemic AKI. PMID- 19129256 TI - TGF-beta upregulation drives tertiary lymphoid organ formation and kidney dysfunction in calcineurin A-alpha heterozygous mice. AB - Calcineurin is an important intracellular signaling molecule which can be inhibited by cyclosporin resulting in immune suppression and nephrotoxicity. Previously, we reported that homozygous loss of the alpha isoform of calcineurin impairs kidney development and function and mimics many features of cyclosporin nephrotoxicity. However, early lethality of null mice prevented further study of renal changes. Alternatively, we examined aged heterozygous (CnAalpha(+/-)) mice. In addition to renal dysfunction and inflammation, we find that CnAalpha(+/-) mice spontaneously develop tertiary lymphoid aggregates in the kidney, small intestine, liver, and lung. Lymphoid aggregates contain both T cells and B cells and exhibited organization suggestive of tertiary lymphoid organs (TLOs). Kidney function and TLO formation were highly correlated suggesting that this process may contribute to nephrotoxicity. Consistent with previous findings, transforming growth factor (TGF)-beta is significantly increased in CnAalpha(+/-) mice. Neutralization of TGF-beta attenuated TLO formation and improved kidney function. In conclusion, we report that haploinsufficiency of CnAalpha causes uregulation of TGF-beta which contributes to chronic inflammation and formation of TLOs. While the process that leads to TLOs formation in transplant allografts is unknown, TLOs are associated with poor clinical prognosis. This study suggests that calcineurin inhibition itself may lead to TLO formation and that TGF-beta may be a novel therapeutic target. PMID- 19129257 TI - Regulation of PTH mRNA stability by the calcimimetic R568 and the phosphorus binder lanthanum carbonate in CKD. AB - Secondary hyperparathyroidism is characterized by increased parathyroid hormone (PTH) mRNA stability that leads to increased PTH mRNA and serum PTH levels. PTH gene expression is reduced by the calcimimetic R568 and the oral phosphorus binder lanthanum carbonate (La). Changes in PTH mRNA stability are regulated by the binding of trans-acting stabilizing and destabilizing factors to a defined cis element in the PTH mRNA 3'-untranslated region (UTR). Adenosine-uridine (AU) binding factor 1 (AUF1) is a PTH mRNA-stabilizing protein, and K-homology splicing regulatory protein (KSRP) is a destabilizing protein that targets mRNAs, including PTH mRNA, to degradation by the ribonuclease complex exosome. We now show that KSRP-PTH mRNA binding is decreased in parathyroids from rats with adenine-induced chronic kidney disease (CKD) where PTH mRNA is more stable. KSRP PTH mRNA binding is increased by treatment with both R568 and La, correlating with decreased PTH gene expression. In vitro degradation assays using transcripts for PTH mRNA and rat parathyroid extracts reproduce the differences in mRNA stability in vivo. Accordingly, PTH mRNA is destabilized in vitro by parathyroid extracts from CKD rats treated with R568 or La compared with parathyroid extracts from untreated CKD rats. This destabilizing effect of R568 and La is dependent on KSRP and the PTH mRNA 3'-UTR. Therefore, the calcimimetic R568 and correction of serum phosphorus by La determine PTH mRNA stability through KSRP-mediated recruitment of a degradation complex to the PTH mRNA, thereby decreasing PTH expression. PMID- 19129258 TI - Glomerular 20-HETE, EETs, and TGF-beta1 in diabetic nephropathy. AB - The early stage of diabetic nephropathy (DN) is linked to proteinuria. Transforming growth factor (TGF)-beta1 increases glomerular permeability to albumin (P(alb)), whereas 20-HETE and EETs reduce P(alb). To investigate the impact of hyperglycemia and hyperlipidemia on 20-HETE, EETs, and TGF-beta1 in the glomeruli, rats were divided into four groups: ND rats were fed a normal diet, HF rats were fed a high-fat diet, STZ rats were treated with 35 mg/kg of streptozotocin, and HF/STZ rats were fed a HF diet and treated with STZ. After 10 wk on these regimens, blood glucose, urinary albumin, serum cholesterol, serum triglyceride levels, and the kidney-to-body weight ratio were significantly elevated in STZ and HF/STZ rats compared with HF and ND rats. STZ and HF/STZ rats had histopathologic changes and abnormal renal hemodynamics. Expression of glomerular CYP4A, enzymes for 20-HETE production, was significantly decreased in STZ rats, whereas expression of glomerular CYP2C and CYP2J, enzymes for EETs production, was significantly decreased in both STZ and HF/STZ rats. Moreover, glomerular TGF-beta1 levels were significantly greater in STZ and HF/STZ rats than in HF and ND rats. Five-week treatment of STZ rats with clofibrate induced glomerular CYP4A expression and 20-HETE production, but reduced glomerular TGF beta1 and urinary protein excretion. These results demonstrate that hyperglycemia increases TGF-beta1 but decreases 20-HETE and EETs production in the glomeruli, changes that may be important in causing glomerular damage in the early stage of DN. PMID- 19129259 TI - Glomerular endothelial cell fenestrations: an integral component of the glomerular filtration barrier. AB - Glomerular endothelial cell (GEnC) fenestrations are analogous to podocyte filtration slits, but their important contribution to the glomerular filtration barrier has not received corresponding attention. GEnC fenestrations are transcytoplasmic holes, specialized for their unique role as a prerequisite for filtration across the glomerular capillary wall. Glomerular filtration rate is dependent on the fractional area of the fenestrations and, through the glycocalyx they contain, GEnC fenestrations are important in restriction of protein passage. Hence, dysregulation of GEnC fenestrations may be associated with both renal failure and proteinuria, and the pathophysiological importance of GEnC fenestrations is well characterized in conditions such as preeclampsia. Recent evidence suggests a wider significance in repair of glomerular injury and in common, yet serious, conditions, including diabetic nephropathy. Study of endothelial cell fenestrations is challenging because of limited availability of suitable in vitro models and by the requirement for electron microscopy to image these sub-100-nm structures. However, extensive evidence, from glomerular development in rodents to in vitro studies in human GEnC, points to vascular endothelial growth factor (VEGF) as a key inducer of fenestrations. In systemic endothelial fenestrations, the intracellular pathways through which VEGF acts to induce fenestrations include a key role for the fenestral diaphragm protein plasmalemmal vesicle-associated protein-1 (PV-1). The role of PV-1 in GEnC is less clear, not least because of controversy over existence of GEnC fenestral diaphragms. In this article, the structure-function relationships of GEnC fenestrations will be evaluated in depth, their role in health and disease explored, and the outlook for future study and therapeutic implications of these peculiar structures will be approached. PMID- 19129260 TI - Hypertension in mice lacking the CXCR3 chemokine receptor. AB - The CXC chemokine receptor 3 (CXCR3) has been linked to autoimmune and inflammatory disease, allograft rejection, and ischemic nephropathy. CXCR3 is expressed on endothelial and smooth muscle cells. Although a recent study posited that antagonizing of CXCR3 function may reduce atherosclerosis, the role of CXCR3 in controlling physiological vascular functions remains unclear. This study demonstrates that disruption of CXCR3 leads to elevated mean arterial pressures in anesthetized and conscious mice, respectively. Stimulation of isolated resistance vessels with various vasoconstrictors showed increased contractibility in CXCR3-/- mice in response to angiotensin II (ANG II) and a decreased vasodilatation in response to acetylcholine (ACh). The increased contractibility was related to higher ANG II type 1 receptor (AT1R) expression, whereas the decreased vasodilatation was related to lower M3-ACh receptor expression in the mesenteric arteries of CXCR3-/- mice compared with wild-type mice. The vasodilatatory response to ACh could be antagonized by the nonselective ACh receptor antagonist atropine and the selective M3 receptor antagonist 4-DAMP, but not by M1, M2, and M4 receptor antagonists. Additionally, EMSA studies revealed that transcription factors SP-1 and EGR-1 interact as a complex with the murine AT1R promoter region. Furthermore, we could show increased expression of SP-1 in CXCR3-/- mice indicating an imbalanced SP-1 and EGR-1 complex formation which causes increased AT1R expression and hypertension. The data indicate that CXCR3 receptor is important in vascular contractility and hypertension, possibly through upregulated AT1R expression. PMID- 19129261 TI - Tubule and cyst formation: lightning strikes twice. PMID- 19129262 TI - Does behaviour modification affect post-stroke risk factor control? Three-year follow-up of a randomized controlled trial. AB - OBJECTIVE: Little is known about the long-term effectiveness after stroke of interventions for behaviour modification and ensuring concordance with therapies. We describe a follow-up study of a previous randomized controlled trial of a brief period of behaviour modification. The aim of this study was to determine outcomes three years after the initial intervention. DESIGN: Survivors of the original cohort were contacted and asked to attend for follow-up interview, within a geriatric day hospital. This study was carried out in the Geriatric Day Hospital at Stobhill Hospital, Balornock Road, Glasgow. INTERVENTIONS: Details of risk factor control, including blood pressure, cholesterol levels and diabetic control, were assessed. Questionnaires used in the initial study were repeated including the Geriatric Depression Scale score, Euroqol Perceived Health Status and Stroke Services Satisfaction Questionnaire. MAIN MEASURES: Primary outcome was collective risk factor control. Clinical outcomes including recurrent cerebrovascular events, medication persistence and perceived health status were also recorded. RESULTS: Mean length of follow-up was 3.6 years (SD 0.43). Of the 205 patients enrolled in the initial study, 102 patients attended for repeat interview(49 intervention/53 control). There were no significant differences in the percentage of controlled risk factors between groups (intervention 51.7% versus control 55.9%, P = 0.53). Similarities were observed in the number of recurrent clinical events and medication persistence between groups. No overall difference was observed in perceived health status, satisfaction with care or depression scores. CONCLUSIONS: Brief intervention with respect to behaviour modification and risk factor control does not appear to have any long-term benefit. These results must be cautiously interpreted in light of the small study number and further research is required. PMID- 19129263 TI - The effects of two different auditory stimuli on functional arm movement in persons with Parkinson's disease: a dual-task paradigm. AB - OBJECTIVE: To examine, in a dual-task paradigm, the effect of auditory stimuli on people with Parkinson's disease. DESIGN: A counterbalanced repeated-measures design. SETTING: A motor control laboratory in a university setting. SUBJECTS: Twenty individuals with Parkinson's disease. EXPERIMENTAL CONDITIONS: Each participant did two experiments (marching music experiment and weather forecast experiment). In each experiment, the participant performed an upper extremity functional task as the primary task and listened to an auditory stimulus (marching music or weather forecast) as the concurrent task. Each experiment had three conditions: listening to the auditory stimulus, ignoring the auditory stimulus and no auditory stimulus. MAIN MEASURES: Kinematic variables of arm movement, including movement time, peak velocity, deceleration time and number of movement units. RESULTS: We found that performances of the participants were similar across the three conditions for the marching music experiment, but were significantly different for the weather forecast experiment. The comparison of condition effects between the two experiments indicated that the effect of weather forecast was (marginally) significantly greater than that of marching music. CONCLUSIONS: The results suggest that the type of auditory stimulus is important to the degree of interference with upper extremity performance in people with Parkinson's disease. Auditory stimuli that require semantic processing (e.g. weather forecast) may distract attention from the primary task, and thus cause a decline in performance. PMID- 19129264 TI - Classification of manual abilities in children with cerebral palsy under 5 years of age: how reliable is the Manual Ability Classification System? AB - OBJECTIVE: To assess the interobserver reliability of the Manual Ability Classification System (MACS) in young children (age 1-5 years) with cerebral palsy. DESIGN: Interobserver reliability study. SETTING: A cross-sectional study of a hospital-based population of children with cerebral palsy. SUBJECTS: Thirty children, 18 boys and 12 girls between 1 and 5 years of age (mean age 2.5 years +/- 14.2 SD, Gross Motor Function Classification System level I-IV). MEASURES: the children were classified by means of the MACS by two independent observers. Interobserver reliability was analysed using Cohen's kappa. RESULTS: Overall interobserver reliability of the MACS for children aged 1-5 years was moderate, with a linear weighted kappa (kappa) of 0.62 (95% confidence interval (CI) 0.49 0.76). According to the generally accepted categories of agreement, reliability was moderate for children under 2 years of age (kappa = 0.55), and good for children between 2 and 5 years of age (kappa = 0.67). CONCLUSION: Classification of manual ability of young children with cerebral palsy is possible between 2 and 5 years of age. For children younger than 2 years old, it should be done with caution. Further development of the MACS for children under 5 years of age is recommended with an emphasis on age-appropriate descriptions of manual abilities. PMID- 19129265 TI - Commentary: C-reactive protein and risk prediction--moving beyond associations to assessing predictive utility and clinical usefulness. PMID- 19129266 TI - Rotating shift work and the metabolic syndrome: a prospective study. AB - BACKGROUND: Several studies have documented on the elevated cardiovascular risk among shift workers. In order to further explore this relation, we aimed at assessing the association between rotating shift work and the incidence of the metabolic syndrome (MetS). METHODS: In this population-based prospective study, 1529 employees from several large Belgian companies were followed for a median observation period of 6.6 years with respect to the onset of the MetS and its separate components. RESULTS: At baseline, 309 men (20.2%) were rotating shift workers. The MetS incidence rate in these shift workers (60.6 per 1000 person years) was increased in comparison with day workers (37.2 per 1000 person-years) with an odds ratio (95% CI) of 1.77 (1.34-2.32). Multivariate adjustment for potential lifestyle and work-related confounders did only marginally affect the strength of the association. The risk for the development of MetS gradually increased independently with accumulated years of shift work. Rotating shift work not only had an impact on MetS as a cluster of conditions but on each of its individual components as well. CONCLUSIONS: Hence, prospective evidence was found that rotating shift work increases the risk for developing the MetS over a period of 6 years. PMID- 19129267 TI - Commentary: trailblazing a research agenda at the interface of pediatrics and genomic discovery--a commentary on the psychological aspects of genomics and child health. AB - Unprecedented advances in human genome science are underway with potential to benefit public health. For example, it is estimated that within a decade, geneticists and epidemiologists will complete a catalog of the majority of genes associated with common chronic diseases. Such rapid advances create possibilities, if not the mandate, for translational research in how best to apply these and other anticipated discoveries for both individual and population health benefit. Driving these discoveries are rapid advances in infrastructure (e.g., the International HapMap Project to catalog human genetic variation; http://www.hapmap.org), analytical methods, and technology. This expansion in capabilities quickly has taken us from a genetics paradigm-where the influence of individual genes on health outcomes is paramount, to a genomics paradigm-where the complex influence of individual genes is considered in concert with each other and with environmental exposures on health outcomes. We discuss these and similar groundbreaking discoveries with an eye toward understanding their importance to child health and human development, and the role of behavioral science research conducted at the interface of pediatrics and genomic discovery. PMID- 19129268 TI - Problem solving and maternal distress at the time of a child's diagnosis of cancer in two-parent versus lone-parent households. AB - OBJECTIVE: To examine negative affectivity and problem-solving abilities for lone mothers and those who are married/partnered subsequent to a child's diagnosis with cancer. METHODS: Negative affectivity and problem-solving strategies were assessed for 464 mothers (87 lone and 377 married/partnered) within 2-16 weeks of their child's diagnosis with cancer. RESULTS: The two groups of mothers did not differ significantly on measures of perceived posttraumatic stress or problem solving; lone mothers reported significantly more symptoms of depression. This difference was no longer significant when maternal education was taken into account. CONCLUSIONS: Negative affectivity and problem-solving abilities were similar for lone mothers and those that are married/partnered shortly after their child has been diagnosed with cancer. Findings are discussed within the context of contemporary strategies to assess marital status as proxy variable for various underlying constructs. PMID- 19129269 TI - Multiple aorto-right cavitary fistula: a rare complication of prosthetic valvular endocarditis in intravenous drug users. AB - Aorto-cavitary fistula is an infrequent complication in infective endocarditis as well as multiple fistula, being the most rare among intracardiac fistulas. The prognosis of infective endocarditis worsens if there are fistulas associated, exceeding the mortality over 40% despite aggressive surgical treatment. Although transoesophageal echocardiography offers a better sensitivity and specificity, sometimes transthoracic echocardiography is enough to obtain these infrequent pictures that we present. PMID- 19129270 TI - Nitric oxide production by the alveolar compartment of the lungs in cirrhotic patients. AB - In cirrhotic patients, alveolar nitric oxide (NO) concentration is increased. This may be secondary to increased output of NO produced by the alveoli (V'(A,NO)) and/or to decreased lung transfer factor of NO. In advanced liver cirrhosis, NO produced by the alveoli may play a role in abnormalities of pulmonary haemodynamics and gas exchanges. In cirrhotic patients, we aimed to measure V'(A,NO) and to compare V'(A,NO) with pulmonary haemodynamics and gas exchange parameters. Measurements were performed in 22 healthy controls and in 29 cirrhotic patients, of whom eight had hepatopulmonary syndrome. Exhaled NO concentrations were measured at multiple expiratory flow rates to derive alveolar NO concentration. V'(A,NO) was the product of alveolar NO concentration by single breath lung transfer factor for NO. V'(A,NO) was increased in patients (median (range) 260 (177-341) nL x min(-1)) compared with controls (79 (60-90), p<0.0001). Alveolar-arterial oxygen tension difference failed to correlate with V'(A,NO). However, cardiac index correlated positively and systemic vascular resistance correlated negatively with V'(A,NO) (r = 0.56, p = 0.001 and r = 0.52, p = 0.004, respectively). In cirrhotic patients, NO was produced in excess by the alveolar compartment of the lungs. Alveolar NO production was associated with hyperdynamic circulatory syndrome but not with arterial oxygenation impairment. PMID- 19129271 TI - Converting venous acid-base and oxygen status to arterial in patients with lung disease. AB - The aim of the present study was to evaluate a method for calculating arterial values of pH, carbon dioxide tension (P(CO(2))) and oxygen tension (P(O(2))) from peripheral venous values. In total, 40 patients were studied. Arterial and peripheral venous blood were sampled at a department of respiratory diseases. Arterial values were calculated from venous, and measured and calculated values of arterial pH, P(CO(2)) and P(O(2)) were compared. Measured and calculated values of pH and P(CO(2)) correlated well, with the difference between them having a very small bias and standard deviation (pH -0.001+/-0.013, P(CO(2)) 0.09+/-0.28 kPa) within those considered acceptable for laboratory equipment and clinical practice. All but four patients had peripheral oxygen saturation (S(p,O(2))) or=20% in seven (33%) patients, fulfilling the commonly used BALF criterion for DAH. There was a trend toward a positive correlation between the percentage of haemosiderin laden macrophages in BALF and parenchymal haemorrhage assessed semiquantitatively by histopathological analysis. Patients with >or=20% haemosiderin-laden macrophages in BALF showed a significantly increased mortality rate (p = 0.047) compared to those with <20%. In patients with an acute onset of diffuse lung infiltrates and respiratory distress, >or=20% haemosiderin-laden macrophages in BALF can occur with DAD, and is not necessarily diagnostic of DAH. The finding of >or=20% haemosiderin-laden macrophages in BALF is associated with a worse prognosis in patients with DAD. PMID- 19129276 TI - Asthma, airway inflammation and epithelial damage in swimmers and cold-air athletes. AB - Endurance athletes show an increased prevalence of airway hyperresponsiveness. The aim of the present study was to evaluate the long-term effects of training on airway responsiveness, inflammation and epithelial damage in swimmers and cold air athletes. In total, 64 elite athletes (32 swimmers and 32 cold-air athletes), 32 mild asthmatic subjects and 32 healthy controls underwent allergy skin prick testing, methacholine challenge and induced sputum analysis. Overall, 69% of swimmers and 28% of cold-air athletes had airway hyperresponsiveness. Sputum neutrophil count correlated with the number of training hours per week in both swimmers and cold-air athletes. Eosinophil counts were higher in swimmers than in healthy subjects, although they were lower than in asthmatic subjects, and correlated with airway hyperresponsiveness in swimmers only. The eosinophil count in cold-air athletes was similar to that in healthy subjects. Bronchial epithelial cell count was not correlated with airway hyperresponsiveness but was significantly increased in swimmers, compared with healthy and asthmatic controls. In conclusion, the present authors observed significant airway inflammation only in competitive athletes with airway hyperresponsiveness. However, the majority of elite athletes showed evidence of bronchial epithelial damage that could possibly contribute to the development of airway hyperresponsiveness. PMID- 19129277 TI - Efficacy of confronting smokers with airflow limitation for smoking cessation. AB - The objective of the present study was to test whether confronting smokers with previously undetected chronic obstructive pulmonary disease (COPD) increases the rate of smoking cessation. In total, 296 smokers with no prior diagnosis of COPD were detected with mild-to-moderate airflow limitation by means of spirometry and randomly allocated to: confrontational counselling by a nurse with nortriptyline for smoking cessation (experimental group); regular counselling by a nurse with nortriptyline (control group 1); or "care as usual" for smoking cessation by the general practitioner (control group 2). Only the experimental group was confronted with their abnormal spirometry (mean forced expiratory volume in one second (FEV(1)) post-bronchodilator 80.5% predicted, mean FEV(1)/forced vital capacity post-bronchodilator 62.5%). There was no difference in cotinine validated prolonged abstinence rate between the experimental group (11.2%) and control group 1 (11.6%) from week 5-52 (odds ratio (OR) 0.96, 95% confidence interval (CI) 0.43-2.18). The abstinence rate was approximately twice as high in the experimental group compared with control group 2 (5.9%), but this difference was not statistically significant (OR 2.02, 95% CI 0.63-6.46). The present study did not provide evidence that the confrontational approach increases the rate of long-term abstinence from smoking compared with an equally intensive treatment in which smokers were not confronted with spirometry. The high failure rates (> or =88%) highlight the need for treating tobacco addiction as a chronic relapsing disorder. PMID- 19129278 TI - Anti-inflammatory effects of salmeterol/fluticasone, tiotropium/fluticasone or tiotropium in COPD. AB - The anti-inflammatory effects of salmeterol/fluticasone (SFP), tiotropium/fluticasone (Tio+FP) and tiotropium (Tio) alone were investigated on the inflammatory cells and mediators in sputum induced from chronic obstructive pulmonary disease patients. Subjects were either newly diagnosed or had not taken any medication for 3 months prior to the study. Subjects (n = 99) were randomised (not double blinded) and received either SFP (100/1,000 microg daily), Tio+FP (18/1,000 microg daily) or Tio (18 microg daily) for 12 weeks. Induced sputum and serum C-reactive protein (CRP) were analysed prior to and at the end of treatment. The results showed that treatment with SFP caused a significant reduction in interleukin (IL)-8 and matrix metalloprotease (MMP)-9 in induced sputum, compared with treatment with Tio alone. There were no treatment differences between the SFP and Tio+FP groups in decreasing IL-8 and MMP-9 levels. The reduction in IL-8 showed significant association with the reduction in MMP-9. All treatment groups failed to significantly reduce the numbers of total cells, neutrophils, macrophages and eosinophils in induced sputum; in addition, there were no treatment differences in terms of improvement of forced expiratory volume in one second, forced vital capacity, CRP or quality of life between the three groups. The anti-inflammatory effects of salmeterol/fluticasone probably contribute to the clinical benefits seen in chronic obstructive pulmonary disease patients. PMID- 19129279 TI - Abnormal mitochondrial function in locomotor and respiratory muscles of COPD patients. AB - Several cellular and molecular alterations have been described in skeletal and respiratory muscles of patients with chronic obstructive pulmonary disease (COPD), but information on potential abnormalities of mitochondrial function is scarce. The aim of the present study was to investigate mitochondrial function in the vastus lateralis (VL) and external intercostalis (EI) of COPD patients. Biopsies from VL and EI were obtained during surgery for lung cancer in 13 patients with mild to moderate COPD (age 68+/-6 yrs, forced expiratory volume in one second (FEV(1)) 66+/-15% predicted) and 19 control subjects (age 67+/-9 yrs, FEV(1) 95+/-18% pred). State 3 and 4 mitochondrial oxygen consumption (V'(O(2),m)), ATP synthesis, citrate synthase, cytochrome oxidase (COX) and complex I-III activities, as well as reactive oxygen species (ROS) production, were determined. In COPD patients, in both muscles, COX activity (VL: COPD 3.0+/ 0.8 versus control 2.0+/-0.8; EI: 3.7+/-1.6 versus 2.4+/-0.9 micromol min(-1) mg( 1)) and ROS production (VL: 1,643+/-290 versus 1,285+/-468; EI: 1,033+/-210 versus 848+/-288 arbitrary units) were increased, whereas state 3 V'(O(2),m) was reduced (VL: 2.9+/-0.3 versus 3.6+/-0.4; EI: 3.6+/-0.3 versus 4.1+/-0.4 mmol min( 1) kg(-1)). Skeletal muscle mitochondria of patients with chronic obstructive pulmonary disease show electron transport chain blockade and excessive production of reactive oxygen species. The concurrent involvement of both vastus lateralis and external intercostalis suggests a systemic (rather than a local) mechanism(s) already occurring in relatively early stages (Global Initiative for Chronic Obstructive Lung Disease stage II) of the disease. PMID- 19129280 TI - Effect of formoterol with or without budesonide in repeated low-dose allergen challenge. AB - The use of combination therapy in mild asthma is debated. The current authors evaluated the effects of formoterol alone and a formoterol/budesonide combination inhaler on asthma deterioration induced by repeated low-dose allergen exposure. In total, 15 subjects with intermittent allergic asthma inhaled low doses of allergen on seven consecutive weekdays in a three-period, crossover, double blind, double-dummy comparison between formoterol 4.5 microg Turbuhaler, budesonide 160 microg/formoterol 4.5 microg Turbuhaler and placebo, each taken as two puffs 30 min after allergen dosing. The outcome variables were: provocative dose of methacholine causing a 20% fall in forced expiratory volume in one second (PD(20)), exhaled nitric oxide fraction (F(eNO)), sputum eosinophils and prostaglandin D(2), and diary card recordings of symptoms (on a scale of 0-10), short-acting beta(2)-agonist use and evening forced expiratory volume in one second (FEV(1)). With placebo treatment, allergen exposure caused significant increases in airway hyperresponsiveness (geometric mean (coefficient of variation) PD(20): 397 (98) microg before versus 168 (82) microg after), F(eNO) (mean+/-sd 46+/-31 ppb before versus 73+/-46 ppb after) and asthma symptom score (mean+/-sd 0.39+/-0.55 before versus 0.68+/-0.67 after). Budesonide/formoterol abolished these changes and significantly improved baseline FEV(1). Formoterol alone, while providing symptom relief, was no better than placebo in protecting against the allergen-induced increase in airway inflammation. Signs of deteriorating asthma, provoked by low-dose allergen, are prevented by short-term use of budesonide/formoterol but not by temporary use of formoterol alone. PMID- 19129281 TI - Pleural plaques and asbestosis: dose- and time-response relationships based on HRCT data. AB - Most studies on asbestos-related diseases are based on chest radiographs, and dose-response relationships are still controversial. The aim of this study was to describe the most relevant parameters of asbestos exposure linked to pleural plaques and asbestosis diagnosed by high-resolution computed tomography (HRCT). A large screening programme including systematic HRCT examinations was organised from 2003 to 2005 in France for formerly asbestos-exposed workers. The time since first exposure (TSFE), level, duration and cumulative exposure to asbestos were used in adjusted unconditional logistic regression to model the relationships of the two diseases. Analysis of a population of 5,545 subjects demonstrated that TSFE (p<0.0001) and cumulative exposure (p = 0.02) (or level, depending on the models used), were independently associated with the frequency of pleural plaques. Only cumulative exposure (p<0.0001) or level of exposure (p = 0.02) were significantly associated with asbestosis. All trend tests were significant for these parameters. To date and to our knowledge, this study is the largest programme based on HRCT for the screening of asbestos-exposed subjects. Both time response and dose-response relationships were demonstrated for pleural plaques, while only dose-response relationships were demonstrated for asbestosis. These parameters must be included in the definition of high-risk populations for HRCT based screening programmes. PMID- 19129282 TI - Characterisation of severe obliterative bronchiolitis in rheumatoid arthritis. AB - The characteristics of patients with rheumatoid arthritis (RA) who develop obliterative bronchiolitis characterised by severe airflow obstruction have been hitherto poorly investigated. A retrospective study of 25 patients with RA and functional evidence of obliterative bronchiolitis (forced expiratory volume in one second (FEV(1))/forced vital capacity (FVC) <50% and/or residual volume (RV)/total lung capacity (TLC) >140% predicted) was conducted. Patients (mean+/ SD age 64+/-11 yrs) included 17 never-smokers and eight ex-smokers (10.5+/-5.4 pack-yrs). The diagnosis of RA preceded respiratory symptoms in 88% of cases. Dyspnoea on exertion was present in all patients and bronchorrhea in 44%. High resolution computed tomography findings included: bronchial wall thickening (96%), bronchiectasis (40%), mosaic pattern (40%), centrilobular emphysema (56%), and reticular and/or ground-glass opacities (32%). Pulmonary function tests showed: FEV(1) 41+/-12% pred, FEV(1)/FVC 49+/-14%, FVC 70+/-20% pred, RV 148+/ 68% pred and RV/TLC 142+/-34% pred. Lung biopsy, available in nine patients, demonstrated constrictive, follicular and mixed bronchiolitis. Patients were followed for 48.2+/-49 months. Treatment was poorly effective. Chronic respiratory failure occurred in 40% of patients, and four patients died. Obliterative bronchiolitis associated with rheumatoid arthritis is a severe and under-recognised condition leading to respiratory failure and death in a high proportion of patients. PMID- 19129283 TI - Different respiratory phenotypes are associated with isocyanate exposure in spray painters. AB - Associations have been observed between exposure to isocyanates, consisting mainly of oligomers, and respiratory symptoms and isocyanate specific sensitisation in spray painters. The aim of the present study was to assess associations between isocyanate exposure and more objective respiratory effect measures such as bronchial hyperresponsiveness (BHR), baseline spirometry and exhaled nitric oxide (eNO) in a subset of spray painters. Methacholine challenge and eNO measurements were performed in 229 workers. Questionnaires and blood samples were obtained. Specific immunoglobulin (Ig)E and IgG to hexamethylene di isocyanate were assessed in serum using various assays. Personal exposure was estimated by combining personal task-based inhalatory exposure measurements and time-activity information. Workers with higher isocyanate exposure were more often hyperresponsive (prevalence ratio comparing the 75th versus 25th percentile of exposure 1.8). In addition, significant exposure-related decreased forced expiratory volume in one second (FEV(1)), FEV(1)/forced vital capacity ratio and flow-volume parameters independent of BHR were found. BHR was more prevalent among sensitised workers. This was statistically significant for only IgG ImmunoCAP (Phadia, Uppsala, Sweden) positive workers. eNO was not associated with exposure although slightly elevated eNO levels in specific IgG positive subjects were found. The current study provides evidence that exposure to isocyanate oligomers is related to asthma with bronchial hyperresponsiveness as a hallmark, but also shows independent chronic obstructive respiratory effects resulting from isocyanate exposure. PMID- 19129284 TI - Quality control of DL,CO instruments in global clinical trials. AB - High inter- and intra-laboratory variability exists for the single-breath diffusing capacity of the lung for carbon monoxide (D(L,CO)) test. To detect small changes in diffusing capacity in multicentre clinical trials, accurate measurements are essential. The present study assessed whether regular D(L,CO) simulator testing maintained or improved instrument accuracy and reduced variability in multicentre trials. The 125 pulmonary function testing laboratories that participated in clinical trials for AIR(R) Inhaled Insulin validated and monitored the accuracy of their D(L,CO) measuring devices using a D(L,CO) simulator, which creates known target values for any device. Devices measuring a simulated D(L,CO) different from target by >3 mL.min-1.mmHg(-1) failed testing and were serviced. Device accuracy was assessed over time and with respect to differences in several variables. Initially, 31 (25%) laboratories had a D(L,CO) device that failed simulator testing. After fixing or replacing devices, 124 (99%) laboratories had passing devices. The percentage of failed tests significantly decreased over time. Differences in geographical region, device type, breath-hold time, temperature and pressure were not associated with meaningful differences in D(L,CO) device accuracy. Regular diffusing capacity of the lung for carbon monoxide simulator testing allows pulmonary function testing laboratories to maintain the accuracy of their diffusing capacity measurements, leading to reduced variability across laboratories in multicentre clinical trials. PMID- 19129285 TI - Prevalence and Risk Factors of Allergies in Turkey (PARFAIT): results of a multicentre cross-sectional study in adults. AB - The Prevalence and Risk Factors of Allergies in Turkey (PARFAIT) study was planned to evaluate the prevalence of and risk factors for asthma and allergic diseases in Turkey. The present analysis used data from 25,843 parents of primary school children, obtained from a cross-sectional questionnaire-based study. A total of 25,843 questionnaires from 14 centres were evaluated. In rural areas, the prevalences asthma, wheezing, allergic rhinitis and eczema in males were: 8.5% (95% confidence interval (CI) 7.9-9.1%), 13.5% (95% CI 12.8-14.2%), 17.5% (95% CI 16.7-18.2%) and 10.8% (95% CI 10.2-11.4%), respectively; and in females were: 11.2% (95% CI 10.9-11.8%), 14.7% (95% CI 14.3-15.1%), 21.2% (95% CI 20.4 22.0%) and 13.1% (95% CI 12.4-13.8%), respectively. In urban areas, the corresponding prevalences in males were: 6.2% (95% CI 5.8-6.6%), 10.8% (95% CI 10.3-11.3%), 11.7% (95% CI 11.4-12.0%) and 6.6% (95% CI 6.2-7.0%), respectively; and in females were: 7.5 % (95% CI 7.9-7.1%), 12.0% (95% CI 11.7-12.3%), 17.0% (95% CI 16.4-17.6%) and 7.3% (95% CI 6.9-7.7%), respectively. Having an atopic first-degree relative or any other atopic diseases had significant effects on the prevalence of allergic diseases. Housing conditions, such as living in a shanty type house, visible moulds at home and use of wood or biomass as heating or cooking material were associated with one or more allergic diseases. Although genetic susceptibility is strongly associated, country- and population-based environmental factors may contribute to increased prevalence rates of allergic diseases. PMID- 19129286 TI - Cigarette smoke extract reduces VEGF in primary human airway epithelial cells. AB - Reduced vascular endothelial growth factor (VEGF) has been reported in bronchoalveolar lavage fluid and lungs of severe emphysema patients. Airway epithelial cells (AEC) are exposed to various environmental insults like cigarette smoke and bacterial infections, but their direct effect on VEGF production in well-differentiated primary human AEC remains unclear. The current authors determined the effect of cigarette smoke extract (CSE) alone and in combination with Mycoplasma pneumoniae (Mp) on VEGF production in well differentiated primary normal human bronchial epithelial (NHBE) and small airway epithelial cells (SAEC) in air-liquid interface cultures. Secretion and expression of VEGF were determined by ELISA and real-time RT-PCR, respectively. Cell growth, apoptosis, extracellular signal-regulated kinase (ERK)1/2 and protein kinase (PK)C signalling pathways were evaluated to further dissect VEGF regulation under CSE treatment. CSE significantly reduced VEGF secretion in NHBE and SAEC. In SAEC, Mp alone significantly increased the VEGF, while the presence of CSE attenuated Mp-induced VEGF production. While ERK inhibitor reduced VEGF secretion only in NHBE, a PKC inhibitor significantly decreased VEGF secretion in both NHBE and SAEC. In conclusion, direct cigarette smoke extract exposure significantly reduced vascular endothelial growth factor production in well differentiated primary human airway epithelial cells, in part through modifying extracellular signal-regulated kinase 1/2 and protein kinase C signalling pathways. PMID- 19129287 TI - Indeterminate results of QuantiFERON TB-2G test performed in routine clinical practice. AB - The present authors assessed risk factors that can promote indeterminate results of QuantiFERON TB-2G (QFT-2G; Cellestis Ltd, Carnegie, Australia) tests. The subjects were 704 patients with suspected tuberculosis (TB) and latent TB infection between January 2005 and December 2007. The QFT-2G test and the tuberculin skin test (TST) were performed for all subjects. If the results of the QFT-2G test were indeterminate, the test was repeated within 1 month. In total, 72 (10.2%) patients showed indeterminate results on the QFT-2G test. Indeterminate results were due to positive control failure in 68 (88.9%) patients and negative control failure in four patients. The results of the TST were negative for 64 patients showing indeterminate results, the remaining eight patients showed a positive response to the TST. Indeterminate results were significantly associated with elderly and immunocompromised patients. Lymphocytopaenia and hypoalbuminaemia were significantly associated with indeterminate laboratory findings. When the QFT-2G test was repeated for all patients showing indeterminate results, 12 (16.7%) patients demonstrated determinate results on the subsequent test. Indeterminate results of the QuantiFERON TB-2G test under routine clinical practice are not infrequent. When scoring QuantiFERON TB-2G test results for elderly and immunocompromised patients, one must be careful because the possibility of obtaining determinate results may be low even if the test is repeated. PMID- 19129288 TI - High prevalence of vertebral deformities in COPD patients: relationship to disease severity. AB - Bone mineral density decreases with advancing chronic obstructive pulmonary disease (COPD) severity, but it is not known whether this is reflected in higher fracture rates. The present authors wanted to compare the prevalence of vertebral deformities in COPD patients with those in a population-based reference group to determine whether the number of deformities was related to the severity of COPD and how far the use of oral corticosteroids (OCS) influenced the prevalence of deformities. In the present cross-sectional study of 465 COPD patients and 462 controls, vertebral deformities were found in 31% of the COPD patients and 18% of the controls. In subjects who had never or sporadically used OCS, deformities were found in 29% of the COPD patients and 17% of the controls. In females, the average number of vertebral deformities was almost two-fold when COPD severity increased from Global Initiative of Chronic Obstructive Lung Disease stage II to III. In males, the use of OCS had a small but significant influence. Prevalence of vertebral deformities was significantly higher in chronic obstructive pulmonary disease patients than in the controls. In females, the average number of deformities was related to chronic obstructive pulmonary disease severity even after adjustment for other known risk factors. The difference between patients and controls remained significant even in those who never or sporadically used oral corticosteroids. PMID- 19129289 TI - Proactive integrated care improves quality of life in patients with COPD. AB - Self-management strategies improve a variety of health-related outcomes for patients with chronic obstructive pulmonary disease (COPD). These strategies, however, are primarily designed to improve chronic disease management and have not focused on early detection and early treatment of exacerbations. In COPD, the majority of exacerbations go unreported and treatment is frequently delayed, resulting in worsened outcomes. Therefore, a randomised clinical trial was designed to determine whether integration of self-management education with proactive remote disease monitoring would improve health-related outcomes. A total of 40 Global Initiative for Chronic Obstructive Lung Disease stage 3 or 4 COPD patients were randomised to receive proactive integrated care (PIC) or usual care (UC) over a 3-month period. The primary and secondary outcomes were change in quality of life, measured by the St George's Respiratory Questionnaire (SGRQ), and change in healthcare costs. PIC dramatically improved SGRQ by 10.3 units, compared to 0.6 units in the UC group. Healthcare costs declined in the PIC group by US$1,401, compared with an increase of US$1,709 in the UC group, but this was not statistically significant. PIC uncovered nine exacerbations, seven of which were unreported. Therefore, proactive integrated care has the potential to improve outcomes in chronic obstructive pulmonary disease patients through effects of self-management, as well as early detection and treatment of exacerbations. PMID- 19129290 TI - Inspiratory duty cycle responses to flow limitation predict nocturnal hypoventilation. AB - Upper airway obstruction (UAO) can elicit neuromuscular responses that mitigate and/or compensate for the obstruction. It was hypothesised that flow-limited breathing elicits specific timing responses that can preserve ventilation due to increases in inspiratory duty cycle rather than respiratory rate. By altering nasal pressure during non-rapid eye movement (non-REM) sleep, similar degrees of UAO were induced in healthy males and females (n = 10 each). Inspiratory duty cycle, respiratory rate and minute ventilation were determined for each degree of UAO during non-REM sleep and compared with the baseline nonflow-limited condition. A dose-dependent increase in the inspiratory duty cycle and respiratory rate was observed in response to increasing severity of UAO. Increases in the inspiratory duty cycle, but not respiratory rate, helped to acutely maintain ventilation. Heterogeneity in these responses was associated with variable degrees of ventilatory compensation, allowing for the segregation of individuals at risk for hypoventilation during periods of inspiratory airflow limitation. Upper airway obstruction constitutes a unique load on the respiratory system. The inspiratory duty cycle, but not the respiratory rate, determine the individual's ability to compensate for inspiratory airflow limitation during sleep, and may represent a quantitative phenotype for obstructive sleep apnoea susceptibility. PMID- 19129291 TI - Effect of sildenafil on acrolein-induced airway inflammation and mucus production in rats. AB - Airway inflammation with mucus overproduction is a distinguishing pathophysiological feature of many chronic respiratory diseases. Phosphodiesterase (PDE) inhibitors have shown anti-inflammatory properties. In the present study, the effect of sildenafil, a potent inhibitor of PDE5 that selectively degrades cyclic guanosine 3',5'-monophosphate (cGMP), on acrolein induced inflammation and mucus production in rat airways was examined. Rats were exposed to acrolein for 14 and 28 days. Sildenafil or distilled saline was administered intragastrically prior to acrolein exposure. Bronchoalveolar lavage fluid (BALF) was acquired for cell count and the detection of pro-inflammatory cytokine levels. Lung tissue was examined for cGMP content, nitric oxide (NO) metabolite levels, histopathological lesion scores, goblet cell metaplasia and mucin production. The results suggested that sildenafil pretreatment reversed the significant decline of cGMP content in rat lungs induced by acrolein exposure, and suppressed the increase of lung NO metabolites, the BALF leukocyte influx and pro-inflammatory cytokine release. Moreover, sildenafil pretreatment reduced acrolein-induced Muc5ac mucin synthesis at both mRNA and protein levels, and attenuated airway inflammation, as well as epithelial hyperplasia and metaplasia. In conclusion, sildenafil could attenuate airway inflammation and mucus production in the rat model, possibly through the nitric oxide/cyclic guanosine 3',5'-monophosphate pathway, and, thus, might have a therapeutic potential for chronic airway diseases. PMID- 19129292 TI - First acute haemodynamic study of soluble guanylate cyclase stimulator riociguat in pulmonary hypertension. AB - Pulmonary hypertension (PH) is associated with impaired production of the vasodilator nitric oxide (NO). Riociguat (BAY 63-2521; Bayer Healthcare AG, Wuppertal, Germany) acts directly on soluble guanylate cyclase, stimulating the enzyme and increasing sensitivity to low NO levels. The present study evaluates riociguat safety, tolerability and efficacy in patients with moderate-to-severe PH (pulmonary arterial hypertension, distal chronic thromboembolic PH or PH with mild to moderate interstitial lung disease). The optimal tolerated dose was identified by incremental dosing in four patients with PH; pharmacodynamic and pharmacokinetic parameters were assessed following single-dose administration (2.5 mg or 1 mg) in 10 and five patients with PH, respectively. All subjects (n = 19) were analysed for safety and tolerability. Riociguat had a favourable safety profile at single doses < or =2.5 mg. It significantly improved pulmonary haemodynamic parameters and cardiac index in patients with PH in a dose-dependent manner, to a greater extent than inhaled NO. Although riociguat also had significant systemic effects and showed no pulmonary selectivity, mean systolic blood pressure remained >110 mmHg. The present report is the first to describe the use of riociguat in patients with pulmonary hypertension. The drug was well tolerated and superior to nitric oxide in efficacy and duration. Riociguat, therefore, has potential as a novel therapy for pulmonary hypertension and warrants further investigation. PMID- 19129293 TI - Compliance in sleep apnoea therapy: influence of home care support and pressure mode. AB - Continuous positive airway pressure (CPAP) is an effective treatment for obstructive sleep apnoea syndrome (OSAS) but therapy adherence is often low. The hypothesis that CPAP-adherence and clinical outcomes can be improved by either using an autoadjusting-CPAP (APAP) device or an intensive support was tested. A controlled parallel group study was performed with 100 newly diagnosed OSAS patients, randomised into 4 groups (n = 25 each): standard or intensive support plus either APAP or CPAP. Intensive support included education and monthly home visits for 6 months. Clinical outcome was monitored by polysomnography at CPAP initiation and, after 3 and 9 months, compliance data were downloaded from the CPAP devices. After 9 months, intensively supported patients returned for follow up in 88 versus 68% in the standard-support-group. Daily usage (mean+/-sem 5.7+/ 0.2 for intensive support versus 4.6+/-0.4 h for standard support), percentage of days used (80.4+/-2.8 versus 57.0+/-5.9%) and proportion of individual sleep time (80.6+/-3.2 versus 64.9+/-6.2%) were also higher. There was no significant difference between APAP or CPAP, (daily usage 5.2+/-0.4 versus 5.1+/-0.3 h, percentage of days 67.9+/-5.0 versus 69.2+/-4.9%, proportion of sleep time 72.5+/ 5.0% versus 72.1+/-5.2%, for APAP and CPAP) but retention rate was higher with CPAP. In summary, intensive support after continuous positive airway pressure initiation, rather than the application of autoadjusting-continuous positive airway pressure, increased therapy adherence. PMID- 19129294 TI - Bitter-responsive gustatory neurons in the rat parabrachial nucleus. AB - Bitterness is a distinctive taste sensation, but central coding for this quality remains enigmatic. Although some receptor cells and peripheral fibers are selectively responsive to bitter ligands, central bitter responses are most typical in broadly tuned neurons. Recently we reported more specifically tuned bitter-best cells (B-best) in the nucleus of the solitary tract (NST). Most had glossopharyngeal receptive fields and few projected to the parabrachial nucleus (PBN), suggesting a role in reflexes. To determine their potential contribution to other functions, the present study investigated whether B-best neurons occur further centrally. Responses from 90 PBN neurons were recorded from anesthetized rats. Stimulation with four bitter tastants (quinine, denatonium, propylthiouracil, cycloheximide) and sweet, umami, salty, and sour ligands revealed a substantial proportion of B-best cells (22%). Receptive fields for B best NST neurons were overwhelmingly foliate in origin, but in PBN, about half received foliate and nasoincisor duct input. Despite convergence, most B-best PBN neurons were as selectively tuned as their medullary counterparts and response profiles were reliable. Regardless of intensity, cycloheximide did not activate broadly tuned acid/sodium (AN) neurons but did elicit robust responses in B-best cells. However, stronger quinine activated AN neurons and concentrated electrolytes stimulated B-best cells, suggesting that B-best neurons might contribute to higher-order functions such as taste quality coding but work in conjunction with other cell types to unambiguously signal bitter-tasting ligands. In this ensemble, B-best neurons would help discriminate sour from bitter stimuli, whereas AN neurons might be more important in differentiating ionic from nonionic bitter stimuli. PMID- 19129295 TI - A simple quantitative method for analyzing electrographic status epilepticus in rats. AB - Electrographic status epilepticus (ESE) is a medical emergency consisting of repetitive seizures and may result in death or severe brain damage. Epilepsy can develop following ESE. The properties of ESE (e.g., duration and intensity) are variable, as are the effects of putative therapeutic treatments. Therefore a straightforward method to quantify different components of ESE would be beneficial for both researchers and clinicians. A frequency range close to the gamma band was selected for extraction of seizure-related activity from the EEG. This filtering strategy reduced motion artifacts and other noise sources in the electrophysiological recordings, thus increasing the signal-to-noise ratio of the EEG spike activity. EEG spiking was quantified using an energy operator and modeled by an eighth-order polynomial. In a benzodiazepine-resistant rat model of pilocarpine-induced ESE, the efficacy of various pharmaceutical agents at suppressing ESE was analyzed with this and other methods on data collected for < or =24 h after ESE induction. This approach allows for the objective, quantitative, and rapid assessment of the effects of both short- and long-lasting pharmacological manipulations on ESE and other forms of prolonged repetitive electrical activity. PMID- 19129296 TI - The vestibulo-auricular reflex. AB - The mammalian orienting response to sounds consists of a gaze shift that can be a combination of head and eye movements. In animals with mobile pinnae, the ears also move. During head movements, vision is stabilized by compensatory rotations of the eyeball within the head because of the vestibulo-ocular reflex (VOR). While studying the gaze shifts made by cats to sounds, a previously uncharacterized compensatory movement was discovered. The pinnae exhibited short latency, goal-directed movements that reached their target while the head was still moving. The pinnae maintained a fixed position in space by counter-rotating on the head with an equal but opposite velocity to the head movement. We call these compensatory ear movements the vestibulo-auricular reflex (VAR) because they shared many kinematic characteristics with the VOR. Control experiments ruled out efference copy of head position signals and acoustic tracking (audiokinetic) of the source as the cause of the response. The VAR may serve to stabilize the auditory world during head movements. PMID- 19129297 TI - Bayesian correction for attenuation of correlation in multi-trial spike count data. AB - When correlation is measured in the presence of noise, its value is decreased. In single-neuron recording experiments, for example, the correlation of selectivity indices in a pair of tasks may be assessed across neurons, but, because the number of trials is limited, the measured index values for each neuron will be noisy. This attenuates the correlation. A correction for such attenuation was proposed by Spearman more than 100 yr ago, and more recent work has shown how confidence intervals may be constructed to supplement the correction. In this paper, we propose an alternative Bayesian correction. A simulation study shows that this approach can be far superior to Spearman's, both in accuracy of the correction and in coverage of the resulting confidence intervals. We demonstrate the usefulness of this technology by applying it to a set of data obtained from the frontal cortex of a macaque monkey while performing serial order and variable reward saccade tasks. There the correction results in a substantial increase in the correlation across neurons in the two tasks. PMID- 19129299 TI - Imaging investigation of postmenopausal bleeding. PMID- 19129300 TI - An elderly man who suddenly lost consciousness. PMID- 19129303 TI - US healthcare spending slows but continues to outpace total economic growth. PMID- 19129298 TI - Data-driven significance estimation for precise spike correlation. AB - The mechanisms underlying neuronal coding and, in particular, the role of temporal spike coordination are hotly debated. However, this debate is often confounded by an implicit discussion about the use of appropriate analysis methods. To avoid incorrect interpretation of data, the analysis of simultaneous spike trains for precise spike correlation needs to be properly adjusted to the features of the experimental spike trains. In particular, nonstationarity of the firing of individual neurons in time or across trials, a spike train structure deviating from Poisson, or a co-occurrence of such features in parallel spike trains are potent generators of false positives. Problems can be avoided by including these features in the null hypothesis of the significance test. In this context, the use of surrogate data becomes increasingly important, because the complexity of the data typically prevents analytical solutions. This review provides an overview of the potential obstacles in the correlation analysis of parallel spike data and possible routes to overcome them. The discussion is illustrated at every stage of the argument by referring to a specific analysis tool (the Unitary Events method). The conclusions, however, are of a general nature and hold for other analysis techniques. Thorough testing and calibration of analysis tools and the impact of potentially erroneous preprocessing stages are emphasized. PMID- 19129305 TI - Indian doctors demand better second line treatment for patients with drug resistant tuberculosis. PMID- 19129306 TI - Readmissions as a result of adverse drug reactions in older people. PMID- 19129307 TI - Comorbidity and repeat admission to hospital for adverse drug reactions in older adults: retrospective cohort study. AB - OBJECTIVES: To identify factors that predict repeat admission to hospital for adverse drug reactions (ADRs) in older adults. DESIGN: Population based retrospective cohort study. SETTING: All public and private hospitals in Western Australia. PARTICIPANTS: 28 548 patients aged >or=60 years with an admission for an ADR during 1980-2000 followed for three years using the Western Australian data linkage system. RESULTS: 5056 (17.7%) patients had a repeat admission for an ADR. Repeat ADRs were associated with sex (hazard ratio 1.08, 95% confidence interval 1.02 to 1.15, for men), first admission in 1995-9 (2.34, 2.00 to 2.73), length of hospital stay (1.11, 1.05 to 1.18, for stays >or=14 days), and Charlson comorbidity index (1.71, 1.46 to 1.99, for score >or=7); 60% of comorbidities were recorded and taken into account in analysis. In contrast, advancing age had no effect on repeat ADRs. Comorbid congestive cardiac failure (1.56, 1.43 to 1.71), peripheral vascular disease (1.27, 1.09 to 1.48), chronic pulmonary disease (1.61, 1.45 to 1.79), rheumatological disease (1.65, 1.41 to 1.92), mild liver disease (1.48, 1.05 to 2.07), moderate to severe liver disease (1.85, 1.18 to 2.92), moderate diabetes (1.18, 1.07 to 1.30), diabetes with chronic complications (1.91, 1.65 to 2.22), renal disease (1.93, 1.71 to 2.17), any malignancy including lymphoma and leukaemia (1.87, 1.68 to 2.09), and metastatic solid tumours (2.25, 1.92 to 2.64) were strong predictive factors. Comorbidities requiring continuing care predicted a reduced likelihood of repeat hospital admissions for ADRs (cerebrovascular disease 0.85, 0.73 to 0.98; dementia 0.62, 0.49 to 0.78; paraplegia 0.73, 0.59 to 0.89). CONCLUSIONS: Comorbidity, but not advancing age, predicts repeat admission for ADRs in older adults, especially those with comorbidities often managed in the community. Awareness of these predictors can help clinicians to identify which older adults are at greater risk of admission for ADRs and, therefore, who might benefit from closer monitoring. PMID- 19129308 TI - Short-form Tai Chi improves standing balance of people with chronic stroke. AB - BACKGROUND AND OBJECTIVE: Our previous findings showed that 4 weeks of intensive Tai Chi practice improved standing balance in healthy seniors. This study set out to investigate whether Tai Chi could improve standing balance in subjects with chronic stroke. METHODS: One hundred thirty-six subjects >6 months after stroke were randomly assigned to a control group (n = 62) practicing general exercises or a Tai Chi group (n = 74) for 12 weeks of training. Each week, 1 hour of group practice was supplemented by 3 hours of self-practice. We used a short-form of Tai Chi consisting of 12 forms that require whole-body movements to be performed in a continuous sequence and demands concentration. A blinded assessor examined subjects at baseline, 6 weeks (mid-program), 12 weeks (end-program), and 18 weeks (follow-up). The 3 outcome measures were (1) dynamic standing balance evaluated by the center of gravity (COG) excursion during self-initiated body leaning in 4 directions, (2) standing equilibrium evaluated in sensory challenged conditions, and (3) functional mobility assessed by Timed-up-and-go score. Mixed model repeated-measures analysis of variance was used to examine between-group differences. RESULTS: When compared with the controls, the Tai Chi group showed greater COG excursion amplitude in leaning forward, backward, and toward the affected and nonaffected sides (P < .05), as well as faster reaction time in moving the COG toward the nonaffected side (P = .014) in the end-program and follow-up assessments. The Tai Chi group also demonstrated better reliance on vestibular integration for balance control at end-program (P = .038). However, neither group improved significantly in Timed-up-and-go scores. CONCLUSIONS: Twelve weeks of short-form Tai Chi produced specific standing balance improvements in people with chronic stroke that outlasted training for 6 weeks. PMID- 19129309 TI - Sex offender treatment: consumer satisfaction and engagement in therapy. AB - Convicted sex offenders attending an outpatient treatment program in Connecticut were surveyed about their experiences in therapy, their perceived importance of treatment content, their satisfaction with the help they receive, and their engagement in therapeutic services. There were strong correlations between perceived importance of content items and satisfaction with services. A robust correlation was also found between engagement and satisfaction. Clients rated accountability and victim empathy as the most important components of treatment. Other popular content areas were thinking errors, relapse prevention concepts, uncovering motivations to offend, and controlling deviant arousal. Most sex offenders valued the peer support and confrontation offered by group therapy. Though reduced recidivism is clearly the crucial measure of treatment success, clients who are engaged in the treatment process and develop healthy interpersonal skills by participating in therapy may be less likely to engage in abusive behavior. Implications for practitioners are discussed. PMID- 19129310 TI - Sorbent augmented hemodialysis systems: are we there yet? AB - Recent publications have reintroduced the concept of using sorbent systems to augment the efficiency of the dialysis process, either by making stationary or compact wearable devices to regenerate dialysis fluid or to target larger molecules for removal by direct blood or plasma contact with sorbent particles. Many of the inherent problems associated with older sorbents have been overcome by designing sorbents with improved biocompatibility and potential for removing molecules beyond the limits of conventional dialysis membranes. One system is approved for use in acute renal failure in the United States, but other devices are not approved for use in humans and continue to be tested in animals and humans. A prototype wearable sorbent device under investigation is not yet able to meet acceptable small molecular weight solute removal, and the other sorbent devices that possess the ability to remove unconventional uremic toxins have not been studied sufficiently in dialysis patients to define their role as augmentation devices. That there is a renewal of interest in sorbents in augmentation of dialysis points to the dissatisfaction with current dialysis technology. PMID- 19129311 TI - Age and comorbidities are effect modifiers of gender disparities in renal transplantation. AB - Women have less access to kidney transplantation than men, but the contributions of age and comorbidity to this disparity are largely unknown. We conducted a national cohort study of 563,197 patients with first-onset ESRD between 2000 and 2005. We used multivariate generalized linear models to evaluate both access to transplantation (ATT), defined as either registration for the deceased-donor waiting list or receiving a live-donor transplant, and survival benefit from transplantation, defined as the relative rate of survival after transplantation compared with the rate of survival on dialysis. We compared relative risks (RRs) between women and men, stratified by age categories and the presence of common comorbidities. Overall, women had 11% less ATT than men. When the model was stratified by age, 18- to 45-yr-old women had equivalent ATT to men (RR 1.01), but with increasing age, ATT for women declined dramatically, reaching a RR of 0.41 for those who were older than 75 yr, despite equivalent survival benefits from transplantation between men and women in all age subgroups. Furthermore, ATT for women with comorbidities was lower than that for men with the same comorbidities, again despite similar survival benefits from transplantation. This study suggests that there is no disparity in ATT for women in general but rather a marked disparity in ATT for older women and women with comorbidities. These disparities exist despite similar survival benefits from transplantation for men and women regardless of age or comorbidities. PMID- 19129312 TI - Divergent role of donor dendritic cells in rejection versus tolerance of allografts. AB - Little is known about heart tissue/donor dendritic cells, which play a key role in mounting alloimmune responses. In this report, we focus on three primary features of donor dendritic cells: their generation, their trafficking after transplantation, and their role in regulating tolerance versus rejection. Using transgenic mice as donors of heart allografts enabled us to monitor trafficking of donor dendritic cells after transplantation. Donor dendritic cells rapidly migrated into secondary lymphoid tissues within 3 h of transplantation. We found that the chemokine receptor CX3CR1 regulates the generation of heart tissue dendritic cells constitutively. Compared with wild-type hearts, CX3CR1(-/-) hearts contained fewer dendritic cells, and heart allografts from CX3CR1(-/-) donors survived significantly longer without immunosuppression. Unexpectedly, though, co-stimulatory blockade with anti-CD154 or CTLA4-Ig induced long-term survival for wild-type heart allografts but not for CX3CR1(-/-) heart allografts. Increasing the dendritic cell frequency in CX3CR1(-/-) hearts by treatment with Flt3L restored the anti-CD154-induced prolongation of CX3CR1(-/-) heart allograft survival. Compared with wild-type donors, depleting transgenic donors of dendritic cells before heart transplantation also markedly worsened chronic rejection under anti-CD154 treatment. These data indicate the importance of the CX3CR1 pathway in the generation of heart tissue dendritic cells and the divergent role of tissue/dendritic cells in rejection versus tolerance. PMID- 19129313 TI - Developmental origins of renal disease: should nephron protection begin at birth? PMID- 19129315 TI - Egg on the table. PMID- 19129316 TI - Must health literacy be a prerequisite for kidney transplantation? PMID- 19129318 TI - Cinacalcet use patterns and effect on laboratory values and other medications in a large dialysis organization, 2004 through 2006. AB - BACKGROUND AND OBJECTIVES: Cinacalcet was introduced in mid-2004 to treat secondary hyperparathyroidism in dialysis patients. We aimed to characterize adult patients who received cinacalcet prescriptions and to determine (1) dosage titration and effects on laboratory values, active intravenous vitamin D use, and phosphate binder prescriptions and (2) percentage who achieved National Kidney Foundation Kidney Disease Outcomes Quality Initiative targets for serum parathyroid hormone, calcium, and phosphorus and experienced biochemical adverse effects. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This observational study evaluated 45,487 prevalent patients from a dialysis organization database linked with the Centers for Medicare and Medicaid Services End-Stage Renal Disease database. Patient characteristics, laboratory values (albumin, parathyroid hormone, calcium, phosphorus), intravenous vitamin D, and oral medication (cinacalcet, phosphate binders) prescriptions were evaluated for cinacalcet patients. RESULTS: By June 2006, almost 32% of patients had received cinacalcet prescriptions. Mean baseline corrected calcium was 9.8 mg/dl and phosphorus was 6.3 mg/dl, and median parathyroid hormone was 577 pg/ml, versus 9.5 mg/dl, 5.3 mg/dl, and 215 pg/ml, respectively, for noncinacalcet patients. Patients with cinacalcet prescriptions for > or =6 mo had corrected calcium reduced by 4.2%, phosphorus by 7.0%, and parathyroid hormone by 29.9% by 12 mo. More cinacalcet patients attained Kidney Disease Outcomes Quality Initiative targets with less hyperparathyroidism, hypercalcemia, and hyperphosphatemia but more hypoparathyroidism and hypocalcemia. Over 12 mo, vitamin D use and use consistency increased, phosphate binder dosages increased, and mean cinacalcet daily dosage reached 55 mg. CONCLUSIONS: Patients with cinacalcet prescriptions exhibited more severe hyperparathyroidism and hyperphosphatemia than noncinacalcet patients. Positive effects were less dramatic than in Phase III clinical trials, possibly as a result of modest, slow dosage titration. PMID- 19129319 TI - John P. Merrill: the father of nephrology as a specialty. PMID- 19129320 TI - Hepatitis C infection and chronic renal diseases. AB - More than 170 million people worldwide are chronically infected with the hepatitis C virus (HCV), which is responsible for over 1 million deaths resulting from cirrhosis and liver cancers. Extrahepatic manifestations are also relevant and include mixed cryoglobulinemia, lymphoproliferative disorders, and kidney disease. HCV infection is both a cause and a complication of chronic kidney disease, occurring largely in the context of mixed cryoglobulinemia. This infection also represents a major medical and epidemiologic challenge in patients with end-stage renal disease on renal replacement therapy with dialysis or transplantation. In these settings the presence of HCV correlates with higher rates of patient mortality than in HCV-negative subjects on dialysis or undergoing kidney transplant. The major concern is the lack of safe and effective drugs to treat HCV-infected patients with chronic kidney disease. Unfortunately, there are no large-scale clinical trials in this population, especially those receiving renal replacement therapy, so that strong evidence for treatment recommendations is scant. This review article provides the readers with the most recent insights on HCV infection both as cause and complication of chronic kidney disease, discusses pitfalls and limitations of current therapies, and reports on preliminary experience with novel therapeutic agents, as well as directions for future research. PMID- 19129322 TI - The Malaga schizophrenia case-register (RESMA): overview of methodology and patient cohort. AB - BACKGROUND: Little information has become available after psychiatric reforms regarding outcomes of persons with schizophrenia and related disorders cared for in community-based mental health facilities. AIMS: The aim of this study was to determine the consequences of psychiatric services in the users of mental health services in Malaga. METHOD: We describe the cohort and methods involved in the Schizophrenia Case Register (RESMA) in Malaga, Spain. All cases (n = 1,022) were users of public mental health services provided in the catchment area over one year. The majority were male (65%), single (68%), living with their original family (50%), with primary education (41%) and living on disability benefits (52%). RESULTS: Concerning use of services, the majority had out-patient contacts (89%). RESULT: s show a substantial overlap in the use of different services during the study period. CONCLUSION: The Malaga Schizophrenia Case Register provides sociodemographic, clinical and service use information for a large sample of patients with schizophrenia or related disorders. Results obtained from the cohort studied will be instrumental for the follow-up and evaluation of the mental health care reform. PMID- 19129323 TI - Public beliefs about the treatment of schizophrenia and depression in Brazil. AB - BACKGROUND: Little is known about public beliefs concerning the treatment of mental disorders in developing countries. AIMS: To assess preferences on sources of help and treatment and the perception of prognosis for schizophrenia and depression. METHOD: A household survey was carried out in 2002 with a probabilistic sample of 1000 individuals, residents of Sao Paulo city, Brazil, with ages ranging from 18-65. Vignettes in colloquial language describing individuals with schizophrenia and depression (according to DSM-IV and ICD-10) were presented, together with a structured questionnaire with questions about the vignette. RESULTS: Results indicated the same general tendencies for schizophrenia and depression. The public considered the help of lay people and psychologists the most useful. Medical professionals are viewed less favorably. The treatments and activities most recommended are psychotherapy and general activities, such as physical activities and attending church services. Medical treatments are seen as more harmful than helpful. The more important determinants were: identification as mental illness, religion, sex and years of school completed. Prognosis with treatment for both disorders is viewed as favorable. CONCLUSIONS: These results may indicate that the public need to receive more information on the full range of treatments options, especially those related to medical treatment. PMID- 19129321 TI - Assessment of myeloperoxidase and oxidative alpha1-antitrypsin in patients on hemodialysis. AB - BACKGROUND AND OBJECTIVES: The present study assesses the effects of the oxidative stress marker, myeloperoxidase (MPO), and the possible MPO-related oxidative stress marker, oxidative alpha(1)-antitrypsin (oxAT), on carotid intima media thickness (CIMT) and protein-energy wasting (PEW) in patients on hemodialysis (HD). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Blood samples were obtained from 383 patients before HD to measure WBC count, serum albumin, lipids, high-sensitivity C-reactive protein (CRP), alpha(1)-antitrypsin (AT), interleukin-6, oxidative LDL-C, MPO, and oxAT. We assessed both CIMT and the geriatric nutritional risk index (GNRI) in this cross-sectional competitive study. RESULTS: Levels of MPO and oxAT correlated. Myeloperoxidase was associated with max-CIMT, and oxAT correlated with max-CIMT and GNRI. Multivariate linear regression models showed that MPO and oxAT were independent predictors of increasing max-CIMT, whereas oxAT, but not MPO, independently correlated with GNRI. In four combined MPO and oxAT groups classified according to median values, a multinomial logistic regression model showed that high MPO together with high oxAT was independently associated with increased max-CIMT. Moreover, the OR for max-CIMT with positive PEW and high MPO was significantly increased in the four groups with combined MPO and PEW. CONCLUSIONS: High MPO with high oxAT and high MPO with PEW seem to contribute to plaque formation in patients on HD, whereas elevated MPO or oxAT alone might not predict increasing CIMT. In contrast, a high oxAT value seems to be an independent predictor of PEW in patients on HD. PMID- 19129324 TI - Extent and determinants of burden of care in Indian families: a comparison between obsessive-compulsive disorder and schizophrenia. AB - BACKGROUND: Limited data is available for family burden in anxiety disorders such as obsessive-compulsive disorder (OCD), particularly from the developing world where families are the primary source of support for patients. AIM: To compare family burden in patients with OCD and schizophrenia in an Indian setting and to examine the influence of caregivers' demographics, patients' illness severity and disability on family burden. METHOD: This comparative cross-sectional study assessed family burden in key relatives of patients with OCD (n = 50) and schizophrenia (n = 30) respectively. RESULTS: Indian families experience significant degrees of burden in the care of their relatives with OCD and schizophrenia. Key relatives' demographic characteristics did not influence burden severity. Illness severity and patients' disability had a direct positive relationship with perceived family burden. CONCLUSION: Indian families of patients with OCD experience burden comparable to that of families of patients with schizophrenia. There is a need to develop local needs-based support programmes for families of patients with psychiatric disorders in India. PMID- 19129325 TI - Brief multi-disciplinary treatment for torture survivors in Nepal: a naturalistic comparative study. AB - BACKGROUND: Little is known about the effectiveness of treatment for torture survivors in low-income settings. Multi-disciplinary treatment is an often used approach for this target group. AIMS: This study was aimed at examining the effectiveness of brief multi-disciplinary treatment for torture survivors in Nepal. METHODS: A naturalistic comparative design with help-seeking torture survivors and internally displaced persons assigned to a treatment and a comparison group respectively ( n = 192; treatment group n = 111, comparison group n = 81), with baseline measurements on psychiatric symptomatology, disability, and functioning and a five-month follow-up (n = 107; treatment group n = 62; comparison group n = 45), was employed. Intervention consisted of brief psychosocial services, minimal medical services and/or legal assistance. RESULTS: Study groups were generally comparable and non-completers did not significantly differ from completers. The treatment group improved more than the comparison group on somatic symptoms, subjective well-being, disability and functioning, with mostly moderate effect sizes. CONCLUSION: Treatment was moderately effective, with regards to reducing the nonspecific mental health consequences of torture, but disability scores remained high. For clients presenting with more severe mental health problems, other treatments that are realistic in the resource-poor Nepali context need to be sought. PMID- 19129326 TI - Disorders of functions and disorders of capacity in relation to sick leave in mental disorders. AB - BACKGROUND: With the publication of the International Classification of Functioning, Disability and Health (ICF), the description of functional health as opposed to symptoms of illness has gained in importance. AIMS: To study the interrelation between disorders of functions, capacity and participation in mental disorders. METHOD: Two hundred and thirteen psychosomatic inpatients were assessed with the Mini-ICF-Rating for Mental Disorders (Mini-ICF-P), the Symptom Checklist (SCL-90-R) and an interview on sick leave. RESULTS: The highest scores in the Mini-ICF-P were found for 'flexibility' (M = 1.64, SD = 0.94), 'structuring of tasks' (M = 1.12, SD = 1.05) and 'endurance' (M = 1.12, SD = 1.02), the lowest for 'self maintenance' (M = 0.19, SD = 0.44), 'mobility' (M = 0.43, SD = 0.85) and 'adherence to regulations' (M = 0.74, SD = 0.98). The correlation between Mini-ICF-P and SCL-90-R GSI is r = 0.51 (p < 0.001), between sick leave and GSI is r = -0.05 and between sick leave and Mini-ICF-P r = 0.34 ( p < 0.001). CONCLUSION: Disorders of capacity are more closely related to disorders of participation/sick leave than disorders of functions. PMID- 19129327 TI - Long-term outcome of leucotomy on behaviour of people with schizophrenia. AB - BACKGROUND: Prefrontal leucotomy was widely used from the late 1930s to early 1950s as a treatment for disorders involving obsessive agitation. Comparatively few studies of the enduring behavioural effects of such surgery exist, while data on mortality and cognition have been better reported. AIMS: We contrast the psychosocial functioning of older individuals with schizophrenia who had undergone prefrontal leucotomy with two groups of their peers who had not undergone such surgery. METHOD: A total of 87 individuals (one female) with a mean age of 70.3 years (SD = 6.84) were evaluated twice 25 months apart using a standardized rating scale. Twenty of the residents, all with schizophrenia, had undergone prefrontal leucotomy approximately 45 years previously. All diagnoses of schizophrenia were confirmed by multiple psychiatrists using DSM-III criteria at the time of the ratings, which were completed by two care staff who knew the residents well. RESULTS: Repeated measures comparisons with schizophrenia and non schizophrenia patient groups showed no significant differences between the leucotomy and unoperated comparison groups on four of the five Multidimensional Observation Scale for Elderly Subjects (MOSES) scales. CONCLUSIONS: These results are consistent with reports of compromised function among individuals who had undergone leucotomy and contrast with some reports of positive changes in behaviour. PMID- 19129328 TI - Effects of safety behaviour on the maintenance of anxiety and negative belief social anxiety disorder. AB - BACKGROUND: Safety behaviour plays an important role in the maintenance of social anxiety disorder (SAD). SAD patients engage in various safety behaviours in social situations in order to decrease the risk of negative evaluations from others. AIMS: The present study examined the effect of safety behaviour on the maintenance of anxiety and negative belief in SAD by using Structural Equation Modelling (SEM). METHODS: Participants were a healthy group (442) and a SAD group (46) who met the SAD criteria for DSM-IV and who had high scores of SAD symptoms. In the assumed maintenance model, independence variables were safety and avoidance behaviour and dependence variables were anxiety and negative belief. RESULTS: This result showed that the SAD group significantly has more high scores than the healthy group in all scales of anxiety, negative belief and avoidance behaviour, expect for safety behaviour. The result of the multiple-group procedure indicated that safety behaviour contributes more strongly to anxiety and negative belief in the SAD group than in the healthy group. CONCLUSIONS: It is speculated that the SAD group have a stronger link between safety behaviour and negative belief than the healthy group, whereas frequency of the use of safety behaviour is equivalent between two groups. These results support the findings of previous studies. PMID- 19129329 TI - The Camberwell elderly mentally ill and their needs for services. AB - BACKGROUND: Previous studies on the elderly mentally ill (graduates) have been undertaken in mental hospital settings and on populations being resettled from hospitals. This paper aims to assess the characteristics and service needs of an epidemiological sample of elderly mentally ill. AIMS: The aim of this study was to identify the characteristics, problems, service utilization and needs of a sample of elderly patients with functional psychosis in a defined epidemiological area. METHOD: Data collected by PRiSM on psychotic patients who lived in two districts of Maudsley Hospital's catchment area were analyzed using the characteristics, problems and the needs for mental health services of those patients who were over the age of 64. These patients were compared with younger patients using the same data. RESULTS: The elderly patients differed significantly in their characteristics and problems from the younger mentally ill persons. The needs assessment procedure (Camberwell Assessment of Needs, CAN) was less sensitive to physical and psychiatric needs of the elderly as it did not reflect the differences between the two age groups. CONCLUSIONS: The lower rate of schizophrenia in the elderly mentally ill compared to the younger community patients and asylum mentally ill is discussed. The explanation may lie in the natural history of the disorder or more plausibly in the implementation of ;community care policy'. The paper concluded that a needs assessment procedure specifically designed to assess the needs of the elderly is required. PMID- 19129331 TI - The Journal of Health Psychology enters its 14th year of publication. AB - The Journal of Health Psychology enters its 14th year of publication with a few new developments to bring to libraries', authors' and readers' attention. These include changes to the Editorial Board, a new web-based manuscript submission portal and an announcement of a new global special issue on Psychology and Poverty Reduction. Lastly, the content of the current issue is introduced. PMID- 19129332 TI - The impact of marital conflict on health and health care utilization in older couples. AB - This study tested the hypothesis that marital distress has a significant effect on health care utilization for older couples. Results from 536 intact couples in long term marriages showed men in dissatisfying marriages are more likely to utilize health care services; women in satisfying marriages are more likely to use health care services. There were no significant partner effects of marital quality on health problems or health care utilization. The association between husbands' marital quality and wives' health care utilization approached significance. Implications for future research and clinical practice, including gender differences, are discussed. PMID- 19129333 TI - Coping, goal adjustment, and positive and negative affect in definitive infertility. AB - The relationships between coping strategies, goal adjustment and positive and negative affect were studied in 83 definitive involuntary childless people. Self report questionnaires were filled out. The findings suggested that positive ways to handle the childlessness were related to positive affect, while negative ways to deal with the childlessness were related to negative affect. Cognitive coping strategies seemed to have a stronger influence on affect than the behavioral coping strategies. Adjusting the goal to have children seemed to be a fruitful way to cope. These findings suggested that intervention programs should pay attention to both cognitive coping strategies and goal adjustment. PMID- 19129334 TI - The relationship between depression and body dissatisfaction across pregnancy and the postpartum: a prospective study. AB - The overall aim of this study was to examine the relationship between depression and body dissatisfaction across pregnancy and the first 12 months postpartum. During pregnancy, women's (N = 116) perceived attractiveness and strength/fitness remained stable, while feeling fat and salience of weight/shape decreased in late pregnancy. During the postpartum, feeling fat and salience of weight/shape increased. Depression and body dissatisfaction scores were correlated with each other concurrently and across multiple time points. However, in baseline controlled prospective analyses, only a model of greater depression late in pregnancy predicting body dissatisfaction at six weeks postpartum and feeling fat throughout the postpartum was supported. PMID- 19129335 TI - A Q-methodological investigation into the meanings of cigarette consumption. AB - This Q-methodological study identified shared subjective explanations of smoking among non-smokers, current smokers and ex-smokers, to consider whether some representations were protective or facilitated quitting. Four factors were identified: named independent addiction; independent non-addiction; anti-smoking; and social addiction. The first two factors were dominated by current and ex smokers, and the last two by non-smokers. Differences emerged on the use of the 'addiction' concept, the use of smoking as a tool for affect management, the role of image manipulation and the general positive and negative perceptions of smoking. The functional use of the different shared smoking representations is discussed. PMID- 19129336 TI - The role of religious activity and spirituality in the health and well-being of older adults. AB - Older adults completed questionnaires of religiosity, spirituality and health, as well as of the potential mediators of healthy behaviors and social support. Church membership related to potential mediators and positive health; given membership, frequency of attendance contributed less to health. Existential well being was strongly related to all outcome health measures. Regression analyses indicated that spiritual wellbeing and prayer contributed to the prediction of psychological wellbeing, subjective well-being, physical symptoms and depression, even when the contributions of age, gender, healthy behaviors and social support were included. Healthy behaviors and social support operated only as partial mediators of the existential-health effects. PMID- 19129337 TI - Psychosocial predictors of dental anxiety. AB - Building on earlier research on psychosocial predictors of dental anxiety, this study examined the predictive ability of several demographic and personality variables. To accomplish this goal, a combined sample of 89 dental patients and 55 college students completed a self-report survey. Results indicated that greater age, female gender, lower income, greater satisfaction with life, greater thought suppression, and a stronger internal health locus of control predicted higher levels of dental anxiety. Possible strategies for reducing both thought suppression and exaggerated levels of internal health locus of control were discussed. PMID- 19129338 TI - Serum cytokines, mood and sleep after a qigong program: is qigong an effective psychobiological tool? AB - Qigong is an ancient Chinese psychosomatic exercise that integrates movement, breathing and meditation into a single multifaceted practice. The present study was designed to assess the effects of qigong practice on serum cytokines, mood and subjective sleep quality. Experimental participants underwent a qigong training program for one month. Blood samples for the quantification of TNF-alpha and IFN-gamma, and several instruments to assess anxiety and depression symptoms as well as SSQ, were obtained before and after the program. Our findings revealed that while the practice of qigong for one month did not alter serum cytokines, it enhanced psychological well-being, including sleep duration. PMID- 19129339 TI - Validation of the Symptoms of Illness Checklist (SIC) as a tool for health psychology research. AB - We developed the Symptoms of Illness Checklist (SIC) to study psychological influences on physical symptoms of illness. A total of 520 participants completed the SIC and, in some samples, the Salient Stressor Impact Questionnaire, Perceived Stress Scale, Daily Hassles and Uplifts Scale, Derogatis Stress Profile, Life Experiences Survey, and the Symptoms CheckList-90-R. The SIC's test retest, internal reliability, and validity verified by physician ratings, were very good. SIC correlations with diverse stress measures were less inflated than those of other instruments indicating the SIC is a reliable and valid tool to study psychological influences on physical illness. PMID- 19129340 TI - HIV/STI risk communication: the effects of scenario-based risk information and frequency-based risk information on perceived susceptibility to chlamydia and HIV. AB - Health risk judgments are not merely based on risk statistics but also on the ease with which hypothetical events are imagined. We explored the effects of scenario information as opposed to frequency information on susceptibility perceptions regarding Chlamydia and HIV. Results showed that participants felt more susceptible to Chlamydia after reading frequency information. Scenario information only seemed to affect feelings of susceptibility in participants with no intimate relationship. No effects on perceived susceptibility for HIV were found. Results are discussed in terms of severity of the disease and defensive reactions. PMID- 19129341 TI - Spiritual striving, acceptance coping, and depressive symptoms among adults living with HIV/AIDS. AB - We prospectively examined the effects of spiritual striving, social support, and acceptance coping on changes in depressive symptoms among adults living with HIV/AIDS. Participants were 180 culturally diverse adults with HIV/AIDS. Participants completed measures of spiritual striving, social support, coping styles, and depressive symptoms at baseline, three-month follow-up, and six-month follow-up. A path model showed that spiritual striving had direct and indirect inverse effects on changes in depressive symptoms. The relationship between spiritual striving and depressive symptoms was partially mediated by acceptance coping, but not by social support. Results suggest that people living with HIV/AIDS who strive for spiritual growth in the context of their illness experience less negative affect. PMID- 19129342 TI - Social representations of female orgasm. AB - This study examines women's social representations of female orgasm. Fifty semi structured interviews were conducted with British women. The data were thematically analysed and compared with the content of female orgasm-related writing in two women's magazines over a 30-year period. The results indicate that orgasm is deemed the goal of sex with emphasis on its physiological dimension. However, the women and the magazines graft onto this scientifically driven representation the importance of relational and emotive aspects of orgasm. For the women, particularly those who experience themselves as having problems with orgasm, the scientifically driven representations induce feelings of failure, but are also resisted. The findings highlight the role played by the social context in women's subjective experience of their sexual health. PMID- 19129343 TI - Contextual influences on obstetric morbidity and related care seeking behaviour in Thailand. AB - This study examines the association of individual and community-level characteristics with obstetric morbidity and care seeking behaviour in Thailand. Community characteristics have been recognized to shape individual health, but there have been only a few studies explaining their influence on obstetric morbidity. Multi-level logistic regression analysis among 930 women, living in 86 villages indicated that not the community social and health infrastructure but community impoverishment was associated with the likelihood of reporting the morbidity and seeking appropriate care. The findings suggest that community factors should be taken into consideration while implementing programmes to improve maternal health. PMID- 19129344 TI - Binge-drinking in Britain as a social and cultural phenomenon: the development of a grounded theoretical model. AB - Techniques and procedures of Strauss and Corbin's grounded theory were used to develop a conditional matrix on binge-drinking in Britain, reflecting data of 32 participants, aged between 22 and 58. The core category was 'binge-drinking as a social and cultural phenomenon'. Results show the value of binge-drinking as an enjoyable way of socializing and counter-balancing the demands of daily hassles and routines. Intervening and contextual factors indicate fluctuations in binge drinking behaviour and the social and cultural constraints that may operate, such as 'no drink-driving', 'not when the next day is a work day' and 'not on one's own'. PMID- 19129345 TI - The role of cost and response-efficacy in persuasiveness of health recommendations. AB - The persuasiveness of a health recommendation, among other things, is a function of the cost of engaging in the recommended behavior--such as money, time, effort, and discomfort--and the response-efficacy, defined as the likelihood that adherence to the recommendation would lead to the desired goal. This research investigates how cost and response-efficacy combine when influencing persuasion. Several theories of health behavior view cost and response-efficacy as having independent effects on persuasion, that is, a weighted additive impact. This research posits, and finds empirical support for the idea that cost and efficacy combine in a multiplicative fashion to influence persuasion, and suggests a structural modification to the traditional models of the relationship between cost, response-efficacy, and persuasion. PMID- 19129346 TI - Bald is beautiful?: the psychosocial impact of alopecia areata. AB - This study reviewed previous studies to explore the psychosocial impact of having alopecia areata (AA). Nineteen primary studies, representing a total of 1271 participants from nine different countries are described and the psychosocial impact of AA is considered. Upon review of the evidence, it is apparent that AA causes dramatic and devastating emotions in patients, which can negatively impact their self-esteem, body image, and/or self-confidence. The negative impact of AA may be heightened for female patients due to the societal pressure to be attractive. Treatment for alopecia patients should offer psychological support to foster increased self-esteem and adaptation to their disorder. PMID- 19129347 TI - Physicians' knowledge, attitudes and professional use of RCTs and meta-analyses: a cross-sectional survey. AB - BACKGROUND: Familiarity with Randomized Control Trials (RCTs) and meta-analyses is essential to practice Evidence Based Medicine (EBM). The aims of this study were to describe knowledge, attitudes and professional behavior of physicians towards methods to evaluate the efficacy of health interventions, particularly meta-analysis and to find out their possible associated factors. METHODS: A cross sectional survey was carried out on a random sample of Italian physicians through a self-administered questionnaire. RESULTS: A response rate of 70.1% was achieved (654 questionnaires). Despite satisfactory knowledge and substantial positive attitudes, Italian physicians have not integrated the use of RCTs and meta analyses into their practice to a large extent, because they infrequently read and use RCTs and meta-analyses to make decisions in clinical practice. There is a high correlation between knowledge, attitudes and professional use of RCTs and meta-analyses. Moreover, the results of our multivariate analysis show that the probability of an appropriate professional use, that is higher for doctors who know the English language, have internet access and dedicate a proper amount of time to continuing medical education, increases significantly with a previous exposure to meta-analysis during graduate/post-graduate training (OR 2.25, 95% CI 1.44-3.52), and with the attendance of post-graduate courses about EBM (OR 1.75, 95% CI 1.09-2.82). Finally, Italian physicians demonstrate a high level of interest in further training. CONCLUSIONS: The association between the EBM educational background of doctors and the appropriate professional use of RCTs and meta-analyses suggest that EBM training may promote a more evidence-based practice among physicians. PMID- 19129348 TI - High incidence of severe ischaemic complications in patients with giant cell arteritis irrespective of platelet count and size, and platelet inhibition. AB - OBJECTIVE: Vision loss and ischaemic stroke are feared complications in GCA. We investigated how platelet count and size and platelet inhibition with ASA relate to ischaemic complications in patients with GCA. METHODS: Charts of patients with GCA were retrospectively analysed. Jaw claudication, amaurosis fugax, blurred vision, ischaemic stroke and permanent visual loss were classified as 'ischaemic events'; ischaemic stroke and permanent visual loss were sub-grouped as 'severe ischaemic events'. The incidence of ischaemia and the association to the pre defined covariates age, fever, ESR, platelet count and size and ASA treatment were assessed. RESULTS: Eighty-five patients (mean age 73 yrs, 60% women, 78% biopsy-proven) were included in the analysis. Of the 85 patients, 62 (73%) presented with ischaemic events, 29/85 patients (34%) with severe ischaemic events. At the time of diagnosis 22/85 patients (26%) were treated with ASA. Of these 22 patients, 15 (68%) presented with ischaemic events, 7/22 patients (32%) with severe ischaemic events. In multivariate analysis, neither platelet count nor size or ASA treatment were significantly associated with ischaemic or severe ischaemic events. CONCLUSIONS: The incidence of severe ischaemic events in patients with GCA was high, irrespective of platelet count and size and established ASA treatment. PMID- 19129349 TI - Femoral head osteonecrosis can be caused by disruption of the systemic immune response via the toll-like receptor 4 signalling pathway. AB - OBJECTIVES: Osteonecrosis of the femoral head is observed in patients treated with steroids. However, the pathogenesis of femoral head osteonecrosis remains unclear. We established a rat model with femoral head osteonecrosis by injecting lipopolysaccharide (LPS) and steroid, and assessed the consequences of this on femoral head histology, the systemic immune response and lipid synthesis. METHODS: Male Wistar rats were injected intravenously on days 0 and 1 with 2 mg/kg LPS and intramuscularly with 20 mg/kg methylprednisolone on days 3, 4 and 5. The animals were sacrificed 1, 2, 3 or 4 weeks after the last methylprednisolone injection. Histopathological and biochemical analyses were performed every week. RESULTS: Osteonecrosis of the femoral head was observed in the rats. The plasma triglyceride concentrations had decreased significantly by weeks 2 and 3. The total plasma cholesterol concentrations had increased significantly by week 1 but then decreased significantly by week 4. The plasma concentrations of IL-1beta, IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma and TNF alpha had increased significantly by week 1. These cytokines can all be induced by toll-like receptor 4 (TLR4) signalling. CONCLUSIONS: LPS and methylprednisolone induced osteonecrosis of the femoral head in rats and this was associated with a disruption of the innate immune system and lipid synthesis. These findings suggest that the TLR4 signalling pathway plays an important role in the pathogenesis of femoral head osteonecrosis. PMID- 19129350 TI - Ultrasound colour Doppler measurements in a single joint as measure of disease activity in patients with rheumatoid arthritis--assessment of concurrent validity. AB - OBJECTIVE: Colour Doppler ultrasound (CDU) displays blood flow in the tissues and is able to detect hyperaemia. Because hyperaemia is part of the inflammatory response, the amount of CDU activity in the inflamed synovium may be used to quantify the inflammatory activity in RA. It has never been investigated if the amount of CDU activity in a single joint can be used to quantify disease activity in RA. METHODS: A total of 109 patients with RA and affection of the wrist joint underwent a standardized CDU examination assessing three positions in their most affected wrist at start up in biological treatment. On the same day the following measures of disease activity were collected: assessment of the number of tender and swollen joints, CRP, ESR and 28-joint disease activity score (DAS28). The amount of CDU activity was quantified by measuring the percentage of colour in the synovium--the colour fraction (CF). Correlation between CF and other measures of disease activity was calculated. RESULTS: There was a significant correlation between CF and DAS28 (r = 0.29; P < 0.001), swollen joint count (r = 0.35; P < 0.001), CRP (r = 0.5; P < 0.001) and ESR (r = 0.5; P < 0.001). No other significant correlations were found. CONCLUSION: A standardized ultrasound examination of a single affected wrist joint in patients with RA may be used as a measure of disease activity. More studies are needed to identify the number of joints needed to examine by CDU to obtain the best validity of Doppler measurements. PMID- 19129351 TI - The National Osteoporosis Guideline Group's new guidelines: what is new? PMID- 19129352 TI - Age- and gender-specific normative data of grip and pinch strength in a healthy adult Swiss population. AB - Assessment of hand strength is used in a wide range of clinical settings especially during treatment of diseases affecting the function of the hand. This investigation aimed to determine age- and gender-specific reference values for grip and pinch strength in a normal Swiss population with special regard to old and very old subjects as well as to different levels of occupational demand. Hand strength data were collected using a Jamar dynamometer and a pinch gauge with standard testing position, protocol and instructions. Analysis of the data from 1023 tested subjects between 18 and 96 years revealed a curvilinear relationship of grip and pinch strength to age, a correlation to height, weight and significant differences between occupational groups. Hand strength values differed significantly from those of other populations, confirming the thesis that applying normative data internationally is questionable. Age- and gender specific reference values for grip and pinch strength are presented. PMID- 19129353 TI - An ultrasonographic and anatomical study of carpal tunnel, with special emphasis on the safe zones in percutaneous release. AB - We examined 40 wrists of 12 embalmed and eight fresh cadavers and defined the relative position of the flexor retinaculum to the neurovascular structure, ultrasonographic markers and safe zones by ultrasonography and anatomical dissection. Both longitudinal and transverse ultrasonographic sections clearly depicted the flexor retinaculum, neurovascular bundles, median nerve, flexor tendons and bony boundaries of the underlying joints. Topographic measurement showed [i] good correlation between the actual extent of the flexor retinaculum and the ultrasonographically determined distance between bony landmarks in all hands, and [ii] the widths and lengths of well-defined safe zones. A comparison study confirmed the accuracy of ultrasonography. We conclude that these ultrasonographic landmarks can locate the flexor retinaculum and facilitate safe and complete carpal tunnel release with open or minimally invasive techniques. PMID- 19129354 TI - Use of nerve conduction studies and the pressure-specified sensory device in the diagnosis of carpal tunnel syndrome. AB - Sixty-nine patients with signs of carpal tunnel syndrome (CTS) underwent nerve conduction studies (NCS) and testing with the Pressure-Specified Sensory Device (PSSD). A total of 102 tests were performed (28 bilateral). Twenty patients underwent a carpal tunnel release and were retested after 4 to 6 months. The Symptom Severity Score (SSS) was calculated before and after surgery. A control group of 20 hands in 10 asymptomatic volunteers underwent identical testing. The NCS sensitivity was 87% with a specificity of 90% whereas the PSSD sensitivity was 81% with a specificity of 65%. The combined sensitivity of the two tests was 93%. In the operative group the SSS improved from a mean of 3.34 pre-operatively to 1.95 postoperatively. The NCS improved in 19/21 hands whereas the PSSD improved in 16/19 hands. The non-invasive SSS and PSSD can increase the diagnostic yield in CTS, especially when the NCS are normal. PMID- 19129355 TI - Single injection digital block: is a transthecal injection necessary? AB - Three different methods of injection to obtain digital block anaesthesia were performed on 15 healthy volunteers to evaluate the success and extent of anaesthesia. We found that the traditional transthecal injection technique was inaccurate and the injected agent mainly flowed into the subcutaneous space, and did not remain within the sheath. The deep transthecal single injection kept the anaesthetic agent within the flexor tendon sheath, as intended. The duration of anaesthesia and the area anaesthetised by the subcutaneous injection and the traditional transthecal injection was similar and satisfactory. However, the anaesthetic area after the deep transthecal injection was significantly smaller than that of the other two techniques (P<0.01). A transthecal digital block offers no advantage over a simple subcutaneous digital block. PMID- 19129356 TI - The lateral digital flap for Dupuytren's fasciectomy at the proximal interphalangeal joint--a study of 84 consecutive patients. AB - Fasciectomy is the most common operation performed for Dupuytren's disease. However, as the joint contracture increases, issues of skin deficiency following release become significant. For severe or recurrent disease dermafasciectomy is advised, but a digital transposition flap provides a viable alternative without the need for a skin graft for less-severe disease with moderate skin deficiency. A retrospective review identified 84 patients who had undergone this operation. The flap used was proximally based on the midlateral aspect of the finger allowing direct closure of the donor site. Ninety hands with 134 digits were operated on and 83% of the patients had a full correction. About 70% remained fully corrected at 1 year. All flaps were performed for proximal interphalangeal joint involvement. The mean pre-operative contracture was 34 degrees, corrected postoperatively to 5 degrees. The recurrence rate was 9%. Two percent of the patients had local infection but no flap necrosis was seen. A lateral digital transposition flap is a very stable operation for Dupuytren's disease with moderate skin deficiency and our patients achieved favourable results when compared to other studies that used local flaps combined with skin graft. PMID- 19129357 TI - The effect of rotational malalignment on X-rays of the wrist. AB - This study evaluates the effect of rotational malposition of the wrist on measured radiographic parameters of the distal radius and carpus. Ten wrists from five healthy volunteers were imaged in varying degrees of rotation. The wrists were placed in a custom, calibrated jig and images were taken in 5 degrees increments to a maximum pronation and supination of 20 degrees . The results demonstrate the following: a steady decrease in the scapholunate and radioscaphoid angles with extremes of supination; an increase in palmar tilt with supination and a decrease in palmar tilt with pronation. The palmar tilt varied from -4 degrees to 15 degrees and the scapholunate angle ranged from 48 degrees to 29 degrees at the extremes of rotation. These results demonstrate how malrotation in X-ray positioning can have a significant effect on the apparent alignment of the distal radius and carpal bones. Measurements taken from poor X rays typically fall outside the normal range and could adversely effect treatment decisions. PMID- 19129358 TI - The clinical outcome of scaphoid fracture malunion at 1 year. AB - The aim of this study was to assess the effect of malunion of scaphoid fractures on the clinical outcome at 1 year. Forty-two consecutive patients with united scaphoid waist fractures which had been treated non-operatively underwent longitudinal CT scans to confirm union and assess malunion at 12 to 18 weeks after injury. A blind clinical assessment was made and the Patient Evaluation Measure (PEM) and DASH questionnaires were completed by all the patients 1 year after injury. The group consisted of 38 men and four women with a mean age of 31 years at the time of injury. Correlation analysis revealed no significant relationships between any of the outcome measures (range of motion, grip strength and PEM and DASH scores) and any of the three measures of malunion (height-to length ratio, the dorsal cortical angle and the lateral intra-scaphoid angle). PMID- 19129359 TI - The "open book" flap: a heterodigital cross-finger skin flap and adipofascial flap for coverage of a circumferential soft tissue defect of a digit. AB - A case of circumferential digital skin loss with exposed tendons from the proximal phalanx to the distal interphalangeal joint is presented. This was treated with a two-layer heterodigital cross-finger ("open book") flap from the adjacent digit, utilising a skin-only cross-finger flap to cover the palmar defect and an adipofascial flap to cover the dorsal defect. PMID- 19129360 TI - Sagittal rotational malunions of the distal radius: the role of pure derotational osteotomy. AB - Sagittal rotational malunion after distal radius fractures was identified in eight patients by the presence of a "hinge" point on the volar cortex on the lateral radiograph, and the ulnar head being shorter than the anterior lip of the radius on the posterior-anterior radiograph. The surgical correction consisted of preplating the distal fragment with a volar locking plate before an osteotomy through the "hinge" point, and correcting the dorsal tilt of the distal fragment. Any dorsal defect was filled with cancellous bone graft from the olecranon. Pain, range of motion and grip all improved. Disabilities of arm, shoulder and hand score changed from 54 to six. Dorsal sagittal tilt improved by 26 degrees , from 23 degrees to +3 degrees . Ulnar variance improved by 3 mm, from +1.5 to -1.5 mm, becoming identical to the opposite side. A pure derotational osteotomy corrected the apparent shortening of the radius and restored the volar tilt. PMID- 19129361 TI - Forces transmitted along human flexor tendons--the effect of extending the fingers against the resistance provided by rubber bands. AB - We wished to test the hypothesis that postoperative extension of repaired flexor tendons against rubber bands will reduce the stress on the repairs, and therefore the risk of rupture. During 24 routine carpal tunnel decompression operations the force in flexor tendons was measured using a load cell. The patients flexed and extended their fingers with and without a rubber band providing resistance to extension. We found no statistically significant difference between the force measured in the tendon with or without the presence of rubber bands. To conclude, we have shown that if the application of rubber band dynamic splintage after flexor tendon repair has any advantage, it is not by reducing the forces transmitted along the tendon during resisted extension or by aiding flexion. PMID- 19129362 TI - Four-corner arthrodesis using the Quad memory staple. AB - Range of motion, pain, consolidation and complications were evaluated for nine patients who underwent four-corner arthrodesis using the Quad Memory Staple (QMS) at a mean follow-up of 44 months. The mean pre-operative range of motion was 50 degrees extension, 62 degrees flexion, 9 degrees radial deviation and 24 degrees ulnar deviation. The postoperative range of motion was similar to previous studies at 32 degrees extension, 31 degrees flexion, 15 degrees radial deviation and 20 degrees ulnar deviation. The grip strength was 28 kg pre-operatively and 26 kg postoperatively. The mean pain score improved from 41 to 23 and the Disabilities of Arm, Shoulder, and Hand (DASH) score from 24 to 20. Non-union, haematoma and wound infection were not seen and eventually all four-corner fusions were consolidated. The main advantages of the QMS are its compressive property and the simple fixation technique. It gives good stability, enables early rehabilitation and avoids the risks of pin fixation methods. PMID- 19129363 TI - Two rare cases of association of thumb hypoplasia and polydactyly of the homolateral foot. AB - This paper presents two cases of radial deficiency and thumb aplasia associated with polydactyly of the homolateral foot and a supernumerary toe resembling a thumb. To our knowledge, this association of malformations has not been described previously. PMID- 19129365 TI - Palmar fixation of dorsally displaced distal radius fractures using locking plates with Smartlock locking screws. AB - We performed a prospective cohort study on a consecutive series of 35 unstable, dorsally displaced distal radius fractures, which were treated with palmar locking plates and SmartLock locking screws. There were 17 men and 18 women with a mean age of 44 years who were reviewed at a mean follow-up of 10 months. All the fractures healed at a mean time of 7 weeks. Radiographs did not show any change in alignment from the initial postoperative period until fracture union. The average Mayo wrist score was 90 after fracture union. One patient had an early loss of fracture reduction and one patient developed complex regional pain syndrome. There was no difference in the quality of life before and after surgery. Palmar plate fixation using SmartLock locking screws is effective and safe in stabilising unstable dorsally displaced distal radius fracture and produces good radiological and functional outcomes. PMID- 19129364 TI - The "transverse intraosseous loop technique" (TILT) to re-insert flexor tendons in zone 1. AB - Flexor tendon divisions in Zone 1 need to be re-inserted to the distal phalanx. This paper describes the Transverse Intraosseous Loop Technique (TILT) of re inserting the profundus tendon to the distal phalanx in which an internally placed suture is looped through the bone and tendon interface. It provides a strong repair, which permits the desired tensioning and can be performed in children. Ten of 12 patients' re-insertions using this technique achieved full recovery while two developed fixed flexion contractures. PMID- 19129366 TI - The deep (motor) branch of the ulnar nerve: a detailed examination of its course and the clinical significance of its damage. AB - The deep (motor) branch of the ulnar nerve is not frequently seen during surgery, but is vulnerable to penetrating injury and may be injured during surgery. Its anatomy is poorly described and its three-dimensional configuration is poorly appreciated. In this study, dissections of ten cadaveric specimens have been performed to demonstrate the branching pattern of the nerve and its skeletal relations have been clarified by superimposing images on radiographs. The deep branch passes downwards (anatomical posterior) through a hypothenar fibro muscular tunnel and courses radially on the surface of the interossei. It has four major divisions that are consistent and readily identifiable. Twenty branches were documented supplying hypothenar muscles, medial two lumbricals, interossei, adductor pollicis, all the carpometacarpal and the ring and little metacarpophalangeal joints, and opponens pollicis in 40% and flexor pollicis brevis in 20% of subjects. PMID- 19129367 TI - Metabolomics and its application for non-invasive embryo assessment in IVF. AB - Morphology and cleavage rate remain the mainstay of embryo assessment. However, a number of additional technologies for this application are under investigation. These include the measurement of glucose, lactate, pyruvate or amino acid levels in the embryo culture media, assessment of oxygen consumption by the embryo, genomic and proteomic profiling, and most recently, analytical examination of the embryonic metabolome. As the number of assisted reproduction cycles increases worldwide, improvements in the ability to quickly and non-invasively identify the best embryos for transfer remain a critical goal for reproductive medicine. Recent studies suggest that metabolomic profiling of embryo culture media using optical and non-optical spectroscopies may provide a useful adjunct to the current embryo assessment strategies and provide insight into the phenotype of embryos with increasing reproductive potential. PMID- 19129368 TI - Meiotic maturation failure induced by DICER1 deficiency is derived from primary oocyte ooplasm. AB - RNA interference (RNAi) has diverse functions across cellular processes, including a role in the development of the mammalian oocyte. Mouse primary oocytes deficient in the key RNAi enzyme DICER1 exhibit pronounced defects in chromosome congression and spindle formation during meiotic maturation. The cause of this meiotic maturation failure is unknown. In this study, observations of chromosomes and spindle microtubules during prometaphase in DICER1-deficient oocytes indicate that chromosome congression and spindle formation are overtly normal. Spindle breakdown and chromosome displacement occur after the metaphase plate has formed, during the metaphase to anaphase transition. We hypothesised that this defect could be attributed to either RNAi-mediated regulation of nuclear factors, such as the regulation of centromere chromatin assembly, or the regulation of mRNA expression within the cytoplasm. By transplanting germinal vesicles between DICER1-deficient and wild-type primary oocytes, we show that, unexpectedly, the meiotic failure is not caused by a deficiency derived from the germinal vesicle component. Instead, we reveal that the ooplasm of primary oocytes contains DICER1-dependent factors that are crucial for chromosome segregation and meiotic maturation. PMID- 19129369 TI - Cellular proliferation and vascularization in ovine fetal ovaries: effects of undernutrition and selenium in maternal diet. AB - Sheep were fed a maintenance (M) diet with adequate (A) Se or high (H) Se concentration from 21 days before breeding to day 135 of pregnancy. From day 50 to day 135 of pregnancy (tissue collection day), a portion of the ewes from ASe and HSe groups were fed restricted (R; 60% of M) diet. Fetal ovarian sections were stained for: 1) the presence of proliferating cell nuclear antigen (a marker of proliferating cells) to determine the proportion of proliferating primordial follicles, or the labeling index (LI; percentage of proliferating cells) for primordial, primary, secondary and antral follicles, stromal tissues, and blood vessels; 2) factor VIII (a marker of endothelial cells) or 3) a presence of apoptotic cells/bodies. The number of proliferating primordial follicles and the LI of primordial follicles was decreased by R and/or HSe diets. The LI was similar for theca and granulosa cells, and for secondary or antral follicles, but was greater in secondary and antral than in primordial and primary follicles. R diet and/or Se affected the LI in all follicle types, in stromal tissues and blood vessels. A dense network of blood vessels was detected in the areas containing secondary to antral follicles, medulla, and hilus, but areas containing primordial follicles were poorly vascularized. The number of apoptotic cells was minimal. These results demonstrate that nutrient restriction and/or Se level in the maternal diet affected cellular proliferation in follicles, blood vessels, and stromal tissues in fetal ovaries. Thus, plane of nutrition and Se in the maternal diet may impact fetal ovarian development and function. PMID- 19129370 TI - Effect of dietary-induced changes in plasma insulin concentrations during the early post partum period on pregnancy rate in dairy cows. AB - Dietary stimulation of insulin in post partum dairy cows has been found to enhance ovarian follicle development but to impair oocyte developmental competence. The objective of this study was to test the hypothesis that pregnancy rate would be improved by feeding a diet to stimulate higher insulin (H) until cows resumed ovarian cyclic activity after parturition, and then feeding a diet to lower insulin (L) during the mating period. Each diet was fed to 30 post partum dairy cows until their first rise in milk progesterone, when 15 cows in each group were transferred to the other diet (treatments HL and LH) and 15 cows in each group remained on their original diet (treatments HH and LL) until 120 days post partum. Treatments did not affect dry matter intake, milk yield and metabolisable energy balance. Plasma insulin concentration was elevated in cows fed on H compared with cows fed on L. Treatment did not affect days to first progesterone rise, first oestrus or first insemination. At 120 days post partum, 27% of cows on each of treatments HH, LL and LH were pregnant, but 60% of cows on treatment HL were pregnant (P=0.021). These findings support the concept that physiological relationships between insulin and the reproductive system vary according to stage of the reproductive cycle, and suggest that pregnancy rate can be enhanced by a two-diet strategy tailored to optimise responses before and after the first post partum ovulation. PMID- 19129371 TI - Nuclear factor kappaB pathway and interleukin-6 are affected in eutopic endometrium of women with endometriosis. AB - In order to investigate the role of the nuclear factor kappaB (NFKB) pathway on gene expression in the eutopic endometrium in endometriosis, and in particular of interleukin-6 (IL6), we evaluated RELA, IkappaB kinase (CHUK), NFKBIA and IL6 expressions and NFKB DNA binding in eutopic endometrium from women with endometriosis. Eutopic endometrium was obtained from 37 women with endometriosis and 42 fertile women during laparoscopy. We analysed RELA, CHUK, NFKBIA and IL6 mRNA levels (RT-PCR); RELA, CHUK and NFKBIA proteins and p-NFKBIA/NFKBIA ratio (western blot); and NFKB binding (DNA shift assay) and IL6 concentration (ELISA) in endometrial explants. Our results indicate that mRNA and cytoplasmic proteins of RELA and CHUK exhibit constant levels in normal endometrium during the menstrual cycle. A dramatic increase (P<0.05) in NFKBIA mRNA expression, RELA nuclear presence and the mRNA and the protein of IL6 during late secretory phase was also observed in this tissue. By contrast, in eutopic endometrium from endometriosis patients, a decrease (P<0.05) in IL6 mRNA and protein (61%), NFKBIA mRNA (46%), p-NFKBIA/NFKBIA ratio (42%), RELA nuclear stromal (68%) and CHUK (48%) proteins were found exclusively during the late secretory phase compared with normal endometrium. In conclusion, the canonical activation of NFKB pathway is deregulated and may have reduced transcriptional function affecting NFKBIA and IL6 expression, genes related local proinflammatory processes. These molecular alterations observed during the late secretory phase in eutopic endometrium from endometriosis patients constitute a NFKB system dysfunction, suggesting that NFKB could be an important factor in endometriosis aetiology. PMID- 19129372 TI - Fully human monoclonal antibodies antagonizing the glucagon receptor improve glucose homeostasis in mice and monkeys. AB - Antagonizing the glucagon signaling pathway represents an attractive therapeutic approach for reducing excess hepatic glucose production in patients with type 2 diabetes. Despite extensive efforts, there is currently no human therapeutic that directly inhibits the glucagon/glucagon receptor pathway. We undertook a novel approach by generating high-affinity human monoclonal antibodies (mAbs) to the human glucagon receptor (GCGR) that display potent antagonistic activity in vitro and in vivo. A single injection of a lead antibody, mAb B, at 3 mg/kg, normalized blood glucose levels in ob/ob mice for 8 days. In addition, a single injection of mAb B dose-dependently lowered fasting blood glucose levels without inducing hypoglycemia and improved glucose tolerance in normal C57BL/6 mice. In normal cynomolgus monkeys, a single injection improved glucose tolerance while increasing glucagon and active glucagon-like peptide-1 levels. Thus, the anti GCGR mAb could represent an effective new therapeutic for the treatment of type 2 diabetes. PMID- 19129373 TI - Neurons in the paraventricular nucleus of the hypothalamus inhibit sympathetic outflow to brown adipose tissue. AB - The paraventricular nucleus of the hypothalamus (PVH) plays an important role in energy homeostasis, regulating neuroendocrine, behavioral, and autonomic functions. However, the role of the PVH in regulating thermogenesis and energy expenditure in brown adipose tissue (BAT) is unclear. The present study investigated the effect of activating neurons within the PVH on BAT thermogenesis. In urethane- and chloralose-anesthetized, artificially ventilated rats maintained at a core body temperature of 37.0-38.0 degrees C, microinjection of N-methyl-d-aspartate (NMDA, 12 pmol in 60 nl) in the PVH did not increase BAT sympathetic nerve activity (SNA) or BAT thermogenesis. In contrast, the increase in BAT SNA evoked by body cooling was completely reversed by microinjection of NMDA in the PVH. Additionally, the increases in BAT SNA evoked by body cooling, by microinjection of prostaglandin E(2) (170 pmol in 60 nl) in the medial preoptic area or by microinjection of bicuculline (30 pmol in 60 nl) in the dorsomedial hypothalamus were completely reversed by microinjection of bicuculline (30 pmol in 60 nl) in the PVH. Although the increases in BAT SNA and thermogenesis evoked by microinjection of NMDA (12 pmol in 60 nl) in the raphe pallidus (RPa) was markedly attenuated following microinjection of bicuculline (30 pmol) in the PVH, the increases in BAT SNA and thermogenesis evoked by microinjection of bicuculline (30 pmol in 60 nl) in the RPa were unaffected by microinjection of bicuculline in the PVH. These results demonstrate that disinhibition of neurons in the PVH inhibits BAT SNA likely via activation of a GABAergic input to BAT sympathetic premotor neurons in the RPa. PMID- 19129374 TI - Maternal stress and development of atherosclerosis in the adult apolipoprotein E deficient mouse offspring. AB - Stress is a risk factor for cardiovascular disease, such as atherosclerosis. Stress during pregnancy (maternal stress) may have long-term consequences for the health of the offspring. However, it is not known whether maternal stress affects the offspring's predisposition to develop atherosclerosis. Atherosclerosis is often related to vascular endothelial dysfunction. We hypothesized that maternal stress affects vascular endothelial function and accelerates development of atherosclerosis in offspring of apolipoprotein E-deficient mice, a model commonly used for atherosclerosis research. Stress was induced by restraining dams in small cylinders for five consecutive days (2 h/day) beginning on gestational day 8 +/- 0.5. Vascular function and development of atherosclerosis in the aorta were determined in male and female offspring at 11-15 wk of age (with early lesions) and at 22-26 wk of age (with established lesions). Endothelium-dependent vasorelaxation was determined using methacholine (0.0001-10 micromol/l) in the absence or presence of the nitric oxide synthase inhibitor N(omega)-nitro-l arginine methyl ester hydrochloride (l-NAME; 100 micromol/l). Male offspring (11 15 wk old) from stressed dams were less dependent on nitric oxide for relaxation compared with controls (l-NAME inhibition: 38 +/- 10 vs. 69 +/- 6%, P < 0.05). Atherosclerotic lesion area was larger in male and female 25- to 26-wk-old offspring from stressed dams compared with corresponding controls [median (interquartile range): males: 6.8 (5.4-7.7) vs. 5.1 (4.4-5.5), P < 0.05, females: 10.0 (8.9-10.9) vs. 7.0 (4.7-8.7), P < 0.05]. In conclusion, maternal stress renders the apolipoprotein E-deficient offspring more susceptible to develop atherosclerosis. PMID- 19129375 TI - Role of gut-brain axis in persistent abnormal feeding behavior in mice following eradication of Helicobacter pylori infection. AB - Bacterial infection can trigger the development of functional GI disease. Here, we investigate the role of the gut-brain axis in gastric dysfunction during and after chronic H. pylori infection. Control and chronically H. pylori-infected Balb/c mice were studied before and 2 mo after bacterial eradication. Gastric motility and emptying were investigated using videofluoroscopy image analysis. Gastric mechanical viscerosensitivity was assessed by cardioautonomic responses to distension. Feeding patterns were recorded by a computer-assisted system. Plasma leptin, ghrelin, and CCK levels were measured using ELISA. IL-1beta, TNF alpha, proopiomelanocortin (POMC), and neuropeptide Y mRNAs were assessed by in situ hybridizations on frozen brain sections. Gastric inflammation was assessed by histology and immunohistochemistry. As shown previously, H. pylori-infected mice ate more frequently than controls but consumed less food per bout, maintaining normal body weight. Abnormal feeding behavior was accompanied by elevated plasma ghrelin and postprandial CCK, higher TNF-alpha (median eminence), and lower POMC (arcuate nucleus) mRNA. Infected mice displayed delayed gastric emptying and visceral hypersensitivity. Eradication therapy normalized gastric emptying and improved gastric sensitivity but had no effect on eating behavior. This was accompanied by persistently increased TNF-alpha in the brain and gastric CD3(+) T-cell counts. In conclusion, chronic H. pylori infection in mice alters gastric emptying and mechanosensitivity, which improve after bacterial eradication. A feeding pattern reminiscent of early satiety persists after H. pylori eradication and is accompanied by increased TNF-alpha in the brain. The results support a role for altered gut-brain pathways in the maintenance of postinfective gut dysfunction. PMID- 19129376 TI - Heritable pathologic cardiac hypertrophy in adulthood is preceded by neonatal cardiac growth restriction. AB - The identification of genetic factors influencing cardiac growth independently of increased load is crucial to an understanding of the molecular and cellular basis of pathological cardiac hypertrophy. The central aim of this investigation was to determine how pathological hypertrophy in the adult can be linked with disturbances in cardiomyocyte growth and viability in early neonatal development. The hypertrophic heart rat (HHR) model is derived from the spontaneously hypertensive rat and exhibits marked cardiac hypertrophy, in the absence of a pressure load at maturity. Hearts were harvested from male HHR, and control strain normal heart rats (NHR), at different stages of postnatal development [neonatal (P2), 4 wk, 6 wk, 8 wk, 12 wk, 20 wk]. Isolated neonatal cardiomyocytes were prepared to evaluate cell size, number, and binucleation. At postnatal day 2, HHR hearts were considerably smaller than control NHR (4.3 +/- 0.2 vs. 5.0 +/- 0.1 mg/g, P < 0.05). Cardiac growth restriction in the neonatal HHR was associated with reduced myocyte size (length and width) and an increased proportion of binucleated cardiomyocytes. Furthermore, the number of cardiomyocytes isolated from HHR neonatal hearts was significantly less ( approximately 29%) than NHR. We also observe that growth stress in the neonate is associated with accentuated PI3K and suppressed MAPK activation, although these signaling pathways are normalized in the adult heart exhibiting established hypertrophy. Thus, using the HHR model, we identified novel molecular and cellular mechanisms involving premature exit from the cell cycle, reduced cardiomyocyte endowment, and dysregulated trophic signaling during early development, which are implicated in the etiology of heritable cardiac hypertrophy in the adult. PMID- 19129377 TI - Effects of intracerebroventricular and intra-accumbens melanin-concentrating hormone agonism on food intake and energy expenditure. AB - The brain melanin-concentrating hormone (MCH) system represents an anabolic system involved in energy balance regulation through influences exerted on the homeostatic and nonhomeostatic controls of food intake and energy expenditure. The present study was designed to further delineate the effect of the MCH system on energy balance regulation by assessing the actions of the MCH receptor 1 (MCHR1) agonism on both food intake and energy expenditure after intracerebroventricular (third ventricle) and intra-nucleus-accumbens-shell (intraNAcSH) injections of a MCHR1 agonist. Total energy expenditure and substrate oxidation were assessed following injections in male Wistar rats using indirect calorimetry. Food intake was also measured. Pair-fed groups were added to evaluate changes in thermogenesis that would occur regardless of the meal size and its thermogenic response. Using such experimental conditions, we were able to demonstrate that acute MCH agonism in the brain, besides its orexigenic effect, induced a noticeable change in the utilization of the main metabolic fuels. In pair-fed animals, MCH significantly reduced lipid oxidation when it was injected in the third ventricle. Such an effect was not observed following the injection of MCH in the NAcSH, where MCH nonetheless strongly stimulated appetite. The present results further delineate the influence of MCH on energy expenditure and substrate oxidation while confirming the key role of the NAcSH in the effects of the MCH system on food intake. PMID- 19129378 TI - Toward understanding the neurobiology of social attachment: role of estrogen receptors in the medial amygdala. PMID- 19129379 TI - Microglial VEGF receptor response is an integral chemotactic component in Alzheimer's disease pathology. AB - We hypothesize that microglial chemotactic responses to amyloid-beta peptide (Abeta(1-42)) serve as an early and integral component of inflammatory response in Alzheimer's disease (AD) brain. This study reports a receptor for vascular endothelial growth factor (VEGF), termed VEGF-1 (Flt-1), subserves microglial chemotactic responses induced by Abeta(1-42) stimulation, in vivo and in vitro. Expression of Flt-1 was significantly increased in tissue obtained from AD patients [compared with tissue from nondemented (ND) individuals], in Abeta(1-42) injected rat hippocampus, and in peptide-stimulated human microglia. Single and double immunohistochemical staining demonstrated marked immunoreactivity, for both Flt-1 and its ligand VEGF, in association with microglia and Abeta deposits in AD, but not ND, brain tissue. Functionally, treatment with anti-Flt-1 antibody was highly effective in inhibiting microglial mobility and chemotactic responses measured in vitro using a transwell migration assay. In vivo, transplanted enhanced green fluorescent protein (EGFP)-labeled microglia exhibited Flt-1 dependent chemotaxis induced by peptide injection with anti-Flt-1 effective in blocking migration of cells. Importantly, anti-Flt-1 reduction of microglial mobility was neuroprotective in peptide-injected hippocampus and associated with a significant increase in numbers of viable hippocampal neurons. The results of this study suggest critical functional roles for Flt-1 in mediating microglial chemotactic inflammatory responses which contribute to pathological conditions in AD brain. PMID- 19129380 TI - Increased phospholipase A2 activity and inflammatory response but decreased nerve growth factor expression in the olfactory bulbectomized rat model of depression: effects of chronic ethyl-eicosapentaenoate treatment. AB - An increased inflammatory response and deficient synthesis of neurotrophic factors (NTFs) may contribute to the etiology of depression. However, the interrelationship between inflammation and NTFs is unknown. Recently, ethyl eicosapentaenoate (EPA) has been used to treat depression. The mechanism by which EPA benefits depression is also unclear. Using the olfactory bulbectomized (OB) rat model of depression, this study evaluated two pathways from bulbectomy to the induction of depression-like changes (the inflammation-hypothalamic-pituitary adrenal axis-stress response pathway and inflammation-nerve growth factor-memory pathway) and the effect of EPA on these pathways. When compared with sham operated rats fed a control diet, significantly increased locomotor and rearing activities in an "open field," impaired memory in the Morris water maze, increased expression of corticotrophin-releasing factor (CRF), and increased secretion of corticosterone were found in OB rats. mRNA expression of nerve growth factor (NGF) was significantly lower in the hippocampus, and phospholipase A2 (PLA2) was higher in the hypothalamus; this change was associated with increased interleukin-1beta (IL-1beta) and prostaglandin E2 (PGE2) in the serum and brain. EPA treatments normalized these behavioral impairments and reduced CRF expression and corticosterone secretion. EPA also reduced serum concentrations of IL-1beta and PGE2, but reversed NGF reduction. Similar to the effects of EPA, the anti-inflammatory drug celecoxib significantly reduced blood PGE2, IL-1beta, and corticosterone concentrations and increased NGF expression in OB rats. Furthermore, anti-NGF treatment blocked EPA effects on behavior. These results suggest that an interaction exists between inflammation and NGF in the depression model. EPA may improve depression via its anti-inflammation properties and the upregulation of NGF. PMID- 19129381 TI - Matrix-dependent local retention of secretory vesicle cargo in cortical neurons. AB - Neurons secrete many diffusible signals from synaptic and other secretory vesicles. We characterized secretion of guidance cues, neuropeptides, neurotrophins, and proteases from single secretory vesicles using pHluorin-tagged cargo in cortical neurons. Stimulation triggered transient and persistent fusion events. Transient events represented full release followed by cargo diffusion or incomplete release followed by vesicle retrieval, as previously observed in neuroendocrine cells. Unexpectedly, we also observed that certain cargo, such as Semaphorin 3A (Sema3A), was delivered at the cell surface as stable deposits. Stable deposits and transient events were observed for single cargo and both were SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) and calcium dependent. The ratio between stable and transient events did not depend on cargo size, subcellular localization (synaptic vs extrasynaptic secretion), or the presence of the extracellular matrix. Instead, the ratio is cargo specific and depends on an interaction with the vesicle matrix through a basic domain in the cargo protein. Inhibition of this interaction through deletion of the basic domain in Sema3A abolished stable deposits and rendered all events transient. Strikingly, cargo favoring transient release was stably deposited after corelease with cargo favoring stable deposit. These data argue against cargo diffusion after exocytosis as a general principle. Instead, the vesicle matrix retains secreted signals, probably for focal signaling at the cell surface. PMID- 19129382 TI - Oxytocin makes a face in memory familiar. AB - Social recognition is the basis of all social interactions. Here, we show that, in humans, the evolutionarily highly conserved neuropeptide oxytocin, after intranasal administration, specifically improves recognition memory for faces, but not for nonsocial stimuli. With increased oxytocin levels, previously presented faces were more correctly assessed as "known," whereas the ability of recollecting faces was unchanged. This pattern speaks for an immediate and selective effect of the peptide strengthening neuronal systems of social memory. PMID- 19129383 TI - Personality predicts the brain's response to viewing appetizing foods: the neural basis of a risk factor for overeating. AB - Eating is not only triggered by hunger but also by the sight of foods. Viewing appetizing foods alone can induce food craving and eating, although there is considerable variation in this "external food sensitivity" (EFS). Because increased EFS is associated with overeating, identifying its neural correlates is important for understanding the current epidemic of obesity. Animal research has identified the ventral striatum, amygdala, hypothalamus, medial prefrontal and premotor cortices as key interacting structures for feeding. However, it is unclear whether a similar network exists in humans and how it is affected by EFS. Using functional magnetic resonance imaging, we showed that viewing appetizing compared with bland foods produced changes in connectivity among the human ventral striatum, amygdala, anterior cingulate and premotor cortex that were strongly correlated with EFS. Differences in the dynamic interactions within the human appetitive network in response to pictures of appetizing foods may determine an individual's risk of obesity. PMID- 19129384 TI - Bulbar acetylcholine enhances neural and perceptual odor discrimination. AB - Experimental and modeling data suggest that the circuitry of the main olfactory bulb (OB) plays a critical role in olfactory discrimination. Processing of such information arises from the interaction between OB output neurons local interneurons, as well as interactions between the OB network and centrifugal inputs. Cholinergic input to the OB in particular has been hypothesized to regulate mitral cell odorants receptive fields (ORFs) and behavioral discrimination of similar odorants. We recorded from individual mitral cells in the OB in anesthetized rats to determine the degree of overlap in ORFs of individual mitral cells after exposure to odorant stimuli. Increasing the efficacy of the cholinergic neurotransmission in the OB by addition of the anticholinesterase drug neostigmine (20 mM) sharpened the ORF responses of mitral cells. Furthermore, coaddition of either the nicotinic antagonist methyllycaconitine citrate hydrate (MLA) (20 mM) or muscarinic antagonist scopolamine (40 mM) together with neostigmine (20 mM) attenuated the neostigmine dependent sharpening of ORFs. These electrophysiological findings are predictive of accompanying behavioral experiments in which cholinergic modulation was manipulated by direct infusion of neostigmine, MLA, and scopolamine into the OB during olfactory behavioral tasks. Increasing the efficacy of cholinergic action in the OB increased perceptual discrimination of odorants in these experiments, whereas blockade of nicotinic or muscarinic receptors decreased perceptual discrimination. These experiments show that behavioral discrimination is modulated in a manner predicted by the changes in mitral cell ORFs by cholinergic drugs. These results together present a first direct comparison between neural and perceptual effects of a bulbar neuromodulator. PMID- 19129385 TI - Relating structure to function: Heschl's gyrus and acoustic processing. AB - The way in which normal variations in human neuroanatomy relate to brain function remains largely uninvestigated. This study addresses the question by relating anatomical measurements of Heschl's gyrus (HG), the structure containing human primary auditory cortex, to how this region processes temporal and spectral acoustic information. In this study, subjects' right and left HG were identified and manually indicated on anatomical magnetic resonance imaging scans. Volumes of gray matter, white matter, and total gyrus were recorded, and asymmetry indices were calculated. Additionally, cortical auditory activity in response to noise stimuli varying orthogonally in temporal and spectral dimensions was assessed and related to the volumetric measurements. A high degree of anatomical variability was seen, consistent with other reports in the literature. The auditory cortical responses showed the expected leftward lateralization to varying rates of stimulus change and rightward lateralization of increasing spectral information. An explicit link between auditory structure and function is then established, in which anatomical variability of auditory cortex is shown to relate to individual differences in the way that cortex processes acoustic information. Specifically, larger volumes of left HG were associated with larger extents of rate-related cortex on the left, and larger volumes of right HG related to larger extents of spectral-related cortex on the right. This finding is discussed in relation to known microanatomical asymmetries of HG, including increased myelination of its fibers, and implications for language learning are considered. PMID- 19129386 TI - Laminar specificity of functional input to distinct types of inhibitory cortical neurons. AB - Despite the presence of numerous inhibitory cell types, laminar excitatory input has only been characterized for limited identified types, and it is unknown whether there are differences between cell types in their laminar sources of inhibitory input. In the present study, we characterized sources of local input to nine distinct types of layer 2/3 inhibitory neurons in living slices of mouse somatosensory cortex. Whole-cell recordings from identified cell types, facilitated by use of transgenic mice expressing green fluorescent protein in limited inhibitory neuron populations, were combined with laser scanning photostimulation. We found that each inhibitory cell type received distinct excitatory and inhibitory laminar input patterns. Excitatory inputs could be grouped into three categories. All inhibitory cell types received strong excitation from layer 2/3, and for calretinin (CR)-positive Martinotti cells and burst-spiking interneurons, this was their dominant source of excitatory input. Three other cell types, including fast-spiking basket cells, CR-negative Martinotti cells, and bipolar interneurons, also received strong excitatory input from layer 4. The remaining four inhibitory cell types, including chandelier cells, neurogliaform cells, irregular spiking basket cells, and regular spiking presumptive basket cells, received strong excitatory input from layer 5A and not layer 4. Laminar sources of inhibitory input varied between cell types and could not be predicted from the sources of excitatory input. Thus, there are cell-type specific differences in laminar sources of both excitation and inhibition, and complementary input patterns from layer 4 versus layer 5A suggest cell type differences in their relationships to lemniscal versus paralemniscal pathways. PMID- 19129387 TI - Sensory input enhances synaptogenesis of adult-born neurons. AB - The adult mammalian brain maintains a prominent stem cell niche in the subventricular zone supplying new neurons to the olfactory bulb. We examined the dynamics of synaptogenesis by imaging the formation and elimination of clusters of a postsynaptic marker (PSD95), genetically targeted to adult-born neurons. We imaged in vivo adult-born periglomerular neurons (PGNs) during two phases of development, immaturity and maturity. Immature PGNs showed high levels of PSD95 puncta dynamics during 12-72 h intervals. Mature PGNs were more stable compared with immature PGNs but still remained dynamic, suggesting that synaptogenesis persists long after these neurons integrated into the network. By combining intrinsic signal and two photon imaging we followed PSD95 puncta in sensory enriched glomeruli. Sensory input upregulated the development of adult-born PGNs only in enriched glomeruli. Our data provide evidence for an activity-based mechanism that enhances synaptogenesis of adult-born PGNs during their initial phases of development. PMID- 19129388 TI - Prelude to and resolution of an error: EEG phase synchrony reveals cognitive control dynamics during action monitoring. AB - Error-related activity in the medial prefrontal cortex (mPFC) is thought to work in conjunction with lateral prefrontal cortex (lPFC) as a part of an action monitoring network, where errors signal the need for increased cognitive control. The neural mechanism by which this mPFC-lPFC interaction occurs remains unknown. We hypothesized that transient synchronous oscillations in the theta range reflect a mechanism by which these structures interact. To test this hypothesis, we extracted oscillatory phase and power from current-source-density-transformed electroencephalographic data recorded during a Flanker task. Theta power in the mPFC was diminished on the trial preceding an error and increased immediately after an error, consistent with predictions of an action-monitoring system. These power dynamics appeared to take place over a response-related background of oscillatory theta phase coherence. Theta phase synchronization between FCz (mPFC) and F5/6 (lPFC) sites was robustly increased during error trials. The degree of mPFC-lPFC oscillatory synchronization predicted the degree of mPFC power on error trials, and both of these dynamics predicted the degree of posterror reaction time slowing. Oscillatory dynamics in the theta band may in part underlie a mechanism of communication between networks involved in action monitoring and cognitive control. PMID- 19129389 TI - Cone contacts, mosaics, and territories of bipolar cells in the mouse retina. AB - We report a quantitative analysis of the different bipolar cell types of the mouse retina. They were identified in wild-type mice by specific antibodies or in transgenic mouse lines by specific expression of green fluorescent protein or Clomeleon. The bipolar cell densities, their cone contacts, their dendritic coverage, and their axonal tiling were measured in retinal whole mounts. The results show that each and all cones are contacted by at least one member of any given type of bipolar cell (not considering genuine blue cones). Consequently, each cone feeds its light signals into a minimum of 10 different bipolar cells. Parallel processing of an image projected onto the retina, therefore, starts at the first synapse of the retina, the cone pedicle. The quantitative analysis suggests that our proposed catalog of 11 cone bipolar cells and one rod bipolar cell is complete, and all major bipolar cell types of the mouse retina appear to have been discovered. PMID- 19129390 TI - Loss of modifier of cell adhesion reveals a pathway leading to axonal degeneration. AB - Axonal dysfunction is the major phenotypic change in many neurodegenerative diseases, but the processes underlying this impairment are not clear. Modifier of cell adhesion (MOCA) is a presenilin binding protein that functions as a guanine nucleotide exchange factor for Rac1. The loss of MOCA in mice leads to axonal degeneration and causes sensorimotor impairments by decreasing cofilin phosphorylation and altering its upstream signaling partners LIM kinase and p21 activated kinase, an enzyme directly downstream of Rac1. The dystrophic axons found in MOCA-deficient mice are associated with abnormal aggregates of neurofilament protein, the disorganization of the axonal cytoskeleton, and the accumulation of autophagic vacuoles and polyubiquitinated proteins. Furthermore, MOCA deficiency causes an alteration in the actin cytoskeleton and the formation of cofilin-containing rod-like structures. The dystrophic axons show functional abnormalities, including impaired axonal transport. These findings demonstrate that MOCA is required for maintaining the functional integrity of axons and define a model for the steps leading to axonal degeneration. PMID- 19129391 TI - Distinct memory signatures in the hippocampus: intentional States distinguish match and mismatch enhancement signals. AB - Incoming events that match or mismatch stored representations are thought to influence the ability of the hippocampus to switch between memory encoding and retrieval modes. Electrophysiological work has dissociated match and mismatch signals in the monkey perirhinal cortex, where match signals were selective for matches to goal states, whereas mismatch signals were not modulated by intention (Miller and Desimone, 1994). To investigate whether the theoretically important relational match and mismatch signals in the hippocampus are modulated by goal states, we fully crossed whether a probe stimulus relationally matched or mismatched a previously perceived image or goal state. Subjects performed two working memory tasks in which they either responded "yes" to probes that were identical to the previous sample scene or, after performing a relational manipulation of the scene, responded "yes" only to a probe that was identical to this perceptually novel image. Using functional magnetic resonance imaging, we found evidence for relational match enhancements bilaterally in the hippocampus that were selective for matches between the probe stimulus and goal state, but were not modulated by whether that goal was perceptually novel. Moreover, we found evidence for a complementary hippocampal mismatch enhancement that was triggered by stimuli containing salient perceptual manipulations. Our results provided evidence for parallel memory signatures in the hippocampus: a controlled match signal that can detect matches to internally generated goal states and an automatic mismatch signal that can identify unpredicted perceptual novelty. PMID- 19129393 TI - TRPV1 is activated by both acidic and basic pH. AB - Maintaining physiological pH is required for survival, and exposure to alkaline chemicals such as ammonia (smelling salts) elicits severe pain and inflammation through unknown mechanisms. TRPV1, the capsaicin receptor, is an integrator of noxious stimuli including heat and extracellular acidic pH. Here, we report that ammonia activates TRPV1, TRPA1 (another polymodal nocisensor), and other unknown receptor(s) expressed in sensory neurons. Ammonia and intracellular alkalization activate TRPV1 through a mechanism that involves a cytoplasmic histidine residue, not used by other TRPV1 agonists such as heat, capsaicin or low pH. Our studies show that TRPV1 detects both acidic and basic deviations from homeostatic pH. PMID- 19129392 TI - Intracerebral dendritic cells critically modulate encephalitogenic versus regulatory immune responses in the CNS. AB - Dendritic cells (DCs) appear in higher numbers within the CNS as a consequence of inflammation associated with autoimmune disorders, such as multiple sclerosis, but the contribution of these cells to the outcome of disease is not yet clear. Here, we show that stimulatory or tolerogenic functional states of intracerebral DCs regulate the systemic activation of neuroantigen-specific T cells, the recruitment of these cells into the CNS and the onset and progression of experimental autoimmune encephalomyelitis (EAE). Intracerebral microinjection of stimulatory DCs exacerbated the onset and clinical course of EAE, accompanied with an early T-cell infiltration and a decreased proportion of regulatory FoxP3 expressing cells in the brain. In contrast, the intracerebral microinjection of DCs modified by tumor necrosis factor alpha induced their tolerogenic functional state and delayed or prevented EAE onset. This triggered the generation of interleukin 10 (IL-10)-producing neuroantigen-specific lymphocytes in the periphery and restricted IL-17 production in the CNS. Our findings suggest that DCs are a rate-limiting factor for neuroinflammation. PMID- 19129394 TI - Persistent pain is dependent on spinal mitochondrial antioxidant levels. AB - Reactive oxygen species (ROS) scavengers have been shown to relieve persistent pain; however, the mechanism is not clearly understood. Superoxide produced from mitochondrial oxidative phosphorylation is considered the major source of ROS in neurons during excitation where mitochondrial superoxide levels are normally controlled by superoxide dismutase (SOD-2). The present study hypothesizes that capsaicin-induced secondary hyperalgesia is a consequence of superoxide build-up in spinal dorsal horn neurons and SOD-2 is a major determinant. To test this hypothesis, the spinal levels of SOD-2 activity, inactivated SOD-2 proteins, and mitochondrial superoxide were measured and correlated to the levels of capsaicin induced secondary hyperalgesia in mice with and without SOD-2 manipulations. The data suggest that superoxide accumulation is a culprit in the abnormal sensory processing in the spinal cord in capsaicin-induced secondary hyperalgesia. Our studies also support the notion that SOD-2 nitration is a critical mechanism that maintains elevated superoxide levels in the spinal cord after capsaicin treatment. Finally, our findings suggest a therapeutic potential for the manipulation of spinal SOD-2 activity in pain conditions. PMID- 19129395 TI - Dynamic changes in brain activity during prism adaptation. AB - Prism adaptation does not only induce short-term sensorimotor plasticity, but also longer-term reorganization in the neural representation of space. We used event-related fMRI to study dynamic changes in brain activity during both early and prolonged exposure to visual prisms. Participants performed a pointing task before, during, and after prism exposure. Measures of trial-by-trial pointing errors and corrections allowed parametric analyses of brain activity as a function of performance. We show that during the earliest phase of prism exposure, anterior intraparietal sulcus was primarily implicated in error detection, whereas parieto-occipital sulcus was implicated in error correction. Cerebellum activity showed progressive increases during prism exposure, in accordance with a key role for spatial realignment. This time course further suggests that the cerebellum might promote neural changes in superior temporal cortex, which was selectively activated during the later phase of prism exposure and could mediate the effects of prism adaptation on cognitive spatial representations. PMID- 19129397 TI - Modular control of pointing beyond arm's length. AB - Hand reaching and bipedal equilibrium are two important functions of the human motor behavior. However, how the brain plans goal-oriented actions combining target reaching with equilibrium regulation is not yet clearly understood. An important question is whether postural control and reaching are integrated in one single module or controlled separately. Here, we show that postural control and reaching motor commands are processed by means of a modular and flexible organization. Principal component and correlation analyses between pairs of angles were used to extract global and local coupling during a whole-body pointing beyond arm's length. A low-dimensional organization of the redundant kinematic chain allowing simultaneous target reaching and regulation of the center of mass (CoM) displacement in extrinsic space emerged from the first analysis. In follow-up experiments, both the CoM and finger trajectories were constrained by asking participants to reach from a reduced base of support with or without knee flexion, or by moving the endpoint along a predefined trajectory (straight or semicircular trajectories). Whereas joint covaried during free conditions and under equilibrium restrictions, it was decomposed in two task dependent and task-independent modules, corresponding to a dissociation of arm versus legs, trunk, and head coordination, respectively, under imposed finger path conditions. A numerical simulation supported the idea that both postural and focal subtasks are basically integrated into the same motor command and that the CNS is able to combine or to separate the movement into autonomous functional synergies according to the task requirements. PMID- 19129396 TI - Role of dorsomedial hypothalamic neuropeptide Y in modulating food intake and energy balance. AB - Previous studies have suggested that neuropeptide Y (NPY) in the dorsomedial hypothalamus (DMH) serves as an important signaling peptide in the regulation of energy balance. To elucidate such actions, we used the adenoassociated virus (AAV) system to alter Npy gene expression in the DMH and examined the effects of these alterations on food intake and energy balance as well as explored its downstream signaling pathway. We found that AAV-mediated overexpression of NPY in the DMH of lean rats increased food intake and body weight, and exacerbated high fat diet-induced obesity. Knockdown of NPY expression in the DMH via AAV-mediated RNA interference ameliorated the hyperphagia, obesity, and diabetes of Otsuka Long-Evans Tokushima Fatty (OLETF) rats. NPY knockdown in the DMH produced a nocturnal and meal size-specific feeding effect. Moreover, we found that knockdown of DMH NPY expression in intact rats reduced NPY content in the nucleus of the solitary tract (NTS) and the dorsal motor nucleus of the vagus and affected within-meal satiation. DMH NPY knockdown increased the feeding inhibitory and NTS c-Fos responses to peripheral administration of cholecystokinin. Together, these results indicate that DMH NPY plays an important role in modulating food intake and energy balance and its dysregulation causes disordered energy balance leading to obesity. PMID- 19129398 TI - Homeostatic control of sensory output in basal vomeronasal neurons: activity dependent expression of ether-a-go-go-related gene potassium channels. AB - Conspecific chemosensory communication controls a broad range of social and sexual behaviors. In most mammals, social chemosignals are predominantly detected by sensory neurons of a specialized olfactory subsystem, the vomeronasal organ (VNO). The behavioral relevance of social chemosignaling puts high demands on the accuracy and dynamic range of the underlying transduction mechanisms. However, the physiological concepts implemented to ensure faithful transmission of social information remain widely unknown. Here, we show that sensory neurons in the basal layer of the mouse VNO dynamically control their input-output relationship by activity-dependent regulation of K(+) channel gene expression. Using large scale expression profiling, immunochemistry, and electrophysiology, we provide molecular and functional evidence for a role of ether-a-go-go-related gene (ERG) K(+) channels as key determinants of cellular excitability. Our findings indicate that an increase in ERG channel expression extends the dynamic range of the stimulus-response function in basal vomeronasal sensory neurons. This novel mechanism of homeostatic plasticity in the periphery of the accessory olfactory system is ideally suited to adjust VNO neurons to a target output range in a layer-specific and use-dependent manner. PMID- 19129399 TI - Neurokinin 1 receptors regulate morphine-induced endocytosis and desensitization of mu-opioid receptors in CNS neurons. AB - mu-Opioid receptors (MORs) are G-protein-coupled receptors (GPCRs) that mediate the physiological effects of endogenous opioid neuropeptides and opiate drugs such as morphine. MORs are coexpressed with neurokinin 1 receptors (NK1Rs) in several regions of the CNS that control opioid dependence and reward. NK1R activation affects opioid reward specifically, however, and the cellular basis for this specificity is unknown. We found that ligand-induced activation of NK1Rs produces a cell-autonomous and nonreciprocal inhibition of MOR endocytosis induced by diverse opioids. Studies using epitope-tagged receptors expressed in cultured striatal neurons and a neuroblastoma cell model indicated that this heterologous regulation is mediated by NK1R-dependent sequestration of arrestins on endosome membranes. First, endocytic inhibition mediated by wild-type NK1Rs was overcome in cells overexpressing beta-arrestin2, a major arrestin isoform expressed in striatum. Second, NK1R activation promoted sequestration of beta arrestin2 on endosomes, whereas MOR activation did not. Third, heterologous inhibition of MOR endocytosis was prevented by mutational disruption of beta arrestin2 sequestration by NK1Rs. NK1R-mediated regulation of MOR trafficking was associated with reduced opioid-induced desensitization of adenylyl cyclase signaling in striatal neurons. Furthermore, heterologous regulation of MOR trafficking was observed in both amygdala and locus ceruleus neurons that naturally coexpress these receptors. These results identify a cell-autonomous mechanism that may underlie the highly specific effects of NK1R on opioid signaling and suggest, more generally, that receptor-specific trafficking of arrestins may represent a fundamental mechanism for coordinating distinct GPCR mediated signals at the level of individual CNS neurons. PMID- 19129400 TI - Lipocalin-2 is an autocrine mediator of reactive astrocytosis. AB - Astrocytes, the most abundant glial cell type in the brain, provide metabolic and trophic support to neurons and modulate synaptic activity. In response to a brain injury, astrocytes proliferate and become hypertrophic with an increased expression of intermediate filament proteins. This process is collectively referred to as reactive astrocytosis. Lipocalin 2 (lcn2) is a member of the lipocalin family that binds to small hydrophobic molecules. We propose that lcn2 is an autocrine mediator of reactive astrocytosis based on the multiple roles of lcn2 in the regulation of cell death, morphology, and migration of astrocytes. lcn2 expression and secretion increased after inflammatory stimulation in cultured astrocytes. Forced expression of lcn2 or treatment with LCN2 protein increased the sensitivity of astrocytes to cytotoxic stimuli. Iron and BIM (Bcl-2 interacting mediator of cell death) proteins were involved in the cytotoxic sensitization process. LCN2 protein induced upregulation of glial fibrillary acidic protein (GFAP), cell migration, and morphological changes similar to characteristic phenotypic changes termed reactive astrocytosis. The lcn2-induced phenotypic changes of astrocytes occurred through a Rho-ROCK (Rho kinase)-GFAP pathway, which was positively regulated by nitric oxide and cGMP. In zebrafishes, forced expression of rat lcn2 gene increased the number and thickness of cellular processes in GFAP-expressing radial glia cells, suggesting that lcn2 expression in glia cells plays an important role in vivo. Our results suggest that lcn2 acts in an autocrine manner to induce cell death sensitization and morphological changes in astrocytes under inflammatory conditions and that these phenotypic changes may be the basis of reactive astrocytosis in vivo. PMID- 19129401 TI - Characteristics of responses of Golgi cells and mossy fibers to eye saccades and saccadic adaptation recorded from the posterior vermis of the cerebellum. AB - The anatomical organization of the granular layer of the cerebellum suggests an important function for Golgi cells (GC) in the pathway conveying mossy fiber (MF) afferents to Purkinje cells. Based on such anatomic observations, early proposals have attributed a role in "gain control" for GCs, a function disputed by recent investigations, which assert that GCs instead contribute to oscillatory mechanisms. However, conclusive physiological evidence based on studies of cerebellum-dependent behavior supporting/dismissing the gain control proposition has been lacking as of yet. We addressed the possible function of this interneuron by recording the activity of a large number of both MFs and GCs during saccadic eye movements from the same cortical area of the monkey cerebellum, namely the oculomotor vermis (OMV). Our cellular identification conformed to previously established criteria, mainly to juxtacellular labeling studies correlating physiological parameters with cell morphology. Response patterns of both MFs and GCs were highly heterogeneous. MF discharges correlated linearly with eye saccade metrics and timing, showing directional preference and precise direction tuning. In contrast, GC discharges did not correlate strongly with the metrics or direction of movement. Their discharge properties were also unaffected by motor learning during saccadic adaptation. The OMV therefore receives a barrage of information about eye movements from different oculomotor areas over the MF pathway, which is not reflected in GCs. The unspecificity of GCs has important implications for the intricacies of neuronal processing in the granular layer, clearly discrediting their involvement in gain control and instead suggesting a more secluded role for these interneurons. PMID- 19129402 TI - Astrocyte-derived MCP-1 mediates neuroprotective effects of noradrenaline. AB - The neurotransmitter noradrenaline (NA) can provide neuroprotection against insults including inflammatory stimuli and excitotoxicity, which may involve paracrine effects of neighboring glial cells. Astrocytes express and secrete a variety of inflammatory and anti-inflammatory molecules; however, the effects of NA on astrocyte chemokine expression have not been well characterized. In primary astrocytes, NA increased expression of chemokine CCL2 (MCP-1) at the mRNA and protein levels. NA increased activation of an MCP-1 promoter driving luciferase expression, which was replicated by beta-adrenergic receptor agonists and a cAMP analog, and blocked by a specific beta2-adrenergic receptor antagonist. In primary neurons, addition of MCP-1 reduced NMDA-dependent glutamate release as well as glutamate-dependent Ca(2+) entry. Similarly, conditioned media from NA treated astrocytes reduced glutamate release, an effect that was blocked by neutralizing antibody to MCP-1, whereas MCP-1 dose-dependently reduced neuronal damage attributable to NMDA or to glutamate. MCP-1 significantly reduced lactate dehydrogenase release from neurons after oxygen-glucose deprivation (OGD) and prevented the loss of ATP levels that occurred after OGD or treatment with glutamate. Incubation of neurons with astrocytes separated by a membrane to prevent physical contact showed that NA induced astrocyte release of sufficient MCP-1 to reduce neuronal damage attributable to OGD. These findings indicate that the neuroprotective effects of NA are mediated, at least in part, by induction and release of astrocyte MCP-1. PMID- 19129403 TI - State-, timing-, and pattern-dependent neuromodulation of synaptic strength by a serotonergic interneuron. AB - Here we report that a serotonergic neuron evokes two distinct neuromodulatory actions with different state, timing, and firing pattern dependencies. These neuromodulatory actions may have important behavioral functions. In the mollusc, Tritonia diomedea, EPSCs evoked by ventral swim interneuron B (VSI) exhibited intrinsic plasticity; after a spike train, EPSC amplitude increased from a basal state to a potentiated state, which usually lasted >10 min. While the synapse was in a potentiated state, stimulation of a serotonergic dorsal swim interneuron (DSI) decreased VSI synaptic strength, returning it to a basal state. The extent of the DSI-evoked decrement was strongly correlated with the magnitude of the homosynaptic potentiation. This synaptic reset, or depotentiation, by DSI was blocked by the serotonin receptor antagonist methysergide and mimicked by a serotonin puff. In contrast to this state-dependent neuromodulatory action, we found that a previously described DSI-evoked transient enhancement of VSI synaptic strength was state-independent, producing the same multiplicative increase in EPSC amplitude regardless of whether the synapse was in a potentiated or basal state. These two actions also differed in their dependencies on the firing pattern of DSI and VSI action potentials. Results suggest that state independent synaptic enhancement by DSI may play a short-term role during a swim motor pattern, whereas state-dependent actions may have longer-lasting consequences, resetting VSI synaptic strength after a swim bout. Thus, differences in two neuromodulatory actions at one synapse may allow a serotonergic neuron to play distinct roles at different stages of a motor pattern. PMID- 19129404 TI - Simultaneous silencing of Npy and Dbh expression in hindbrain A1/C1 catecholamine cells suppresses glucoprivic feeding. AB - Previous data have strongly implicated hindbrain catecholamine/neuropeptide Y (NPY) coexpressing neurons as key mediators of the glucoprivic feeding response. Catecholamine/NPY cell bodies are concentrated in the A1 and caudal C1 cell cluster (A1/C1) in the ventrolateral medulla, a region highly sensitive to glucoprivic challenge. To further investigate the importance of this catecholamine subpopulation in glucoregulation, we used small interfering RNA (siRNA) technology to produce a targeted gene knockdown of NPY and dopamine-beta hydroxylase (DBH), a catecholamine biosynthetic enzyme. Unilateral injection of NPY siRNA and DBH siRNA (0.02 nmol each) both significantly inhibited expression of the targeted genes up to 2 d, as revealed by real-time PCR, and reduced protein expression up to 8 d, as revealed by immunohistochemistry, compared with the control nontargeting siRNA (ntRNA) side. Subsequently, targeted siRNA or control ntRNA was injected bilaterally into A1/C1 and responses to 2-deoxy-D glucose (2DG; 200 mg/kg)-induced glucoprivation were tested 3-7 d later. Silencing of either Npy or Dbh alone did not reduce glucoprivic feeding or hyperglycemic responses, compared with responses of ntRNA-injected controls. In contrast, simultaneous silencing of both Npy and Dbh reduced 2DG-induced feeding by 61%. Neither the hyperglycemic response to 2DG nor feeding elicited by mercaptoacetate (68 mg/kg)-induced blockade of fatty acid oxidation ("lipoprivic feeding") was reduced by simultaneous silencing of these two genes. These results suggest that catecholamines and NPY act conjointly to control glucoprivic feeding and that the crucial NPY/catecholamine coexpressing neurons are concentrated in the A1/C1 cell group. PMID- 19129405 TI - Reelin stabilizes the actin cytoskeleton of neuronal processes by inducing n cofilin phosphorylation at serine3. AB - The extracellular matrix protein Reelin, secreted by Cajal-Retzius cells in the marginal zone of the cortex, controls the radial migration of cortical neurons. Reelin signaling involves the lipoprotein receptors apolipoprotein E receptor 2 (ApoER2) and very low density lipoprotein receptor (VLDLR), the adapter protein Disabled1 (Dab1), and phosphatidylinositol-3-kinase (PI3K). Eventually, Reelin signaling acts on the cytoskeleton; however, these effects on cytoskeletal organization have remained elusive. In Reelin-deficient mutant mice, most cortical neurons are unable to migrate to their destinations, suggesting a role for Reelin signaling in the dynamic cytoskeletal reorganization that is required for neurons to migrate. Here, we show that Reelin signaling leads to serine3 phosphorylation of n-cofilin, an actin-depolymerizing protein that promotes the disassembly of F-actin. Phosphorylation at serine3 renders n-cofilin unable to depolymerize F-actin, thereby stabilizing the cytoskeleton. We provide evidence for ApoER2, Dab1, Src family kinases (SFKs), and PI3K to be involved in n-cofilin serine3 phosphorylation. Phosphorylation of n-cofilin takes place in the leading processes of migrating neurons as they approach the Reelin-containing marginal zone. Immunostaining for phospho-cofilin in dissociated reeler neurons is significantly increased after incubation in Reelin-containing medium compared with control medium. In a stripe choice assay, neuronal processes are stable on Reelin-coated stripes but grow on control stripes by forming lamellipodia. These novel findings suggest that Reelin-induced stabilization of neuronal processes anchors them to the marginal zone which appears to be required for the directional migration process. PMID- 19129406 TI - Isolation and characterization of Bartonella quintana from the parotid gland of an immunocompetent man. AB - We describe a case of the isolation of Bartonella quintana from the parotid gland of an apparently healthy man. Pathological examination showed intraparotid granulomatous abscessual lymphadenitis. Diagnosis was made on the basis of high titers of immunoglobulin G (IgG) and IgM antibodies and of culture isolation of a causative agent from parotid aspirate. PMID- 19129407 TI - Unreliability of results of PCR detection of Helicobacter pylori in clinical or environmental samples. AB - The aim of this study was to compare published Helicobacter pylori primer pairs for their ability to reliably detect H. pylori in gastric biopsy specimens and salivary samples. Detection limits of the 26 PCR primer pairs previously described for detection of H. pylori DNA in clinical samples were determined. Sensitivity and specificity were determined using primers with detection limits of <100 CFU/ml using 50 H. pylori-positive and -negative (by concordance by culture and histology) coded gastric biopsy specimens. These results were then confirmed with gastric biopsy specimens and saliva from patients with confirmed H. pylori status. Five of the twenty-six previously reported primer pairs (HP64 f/HP64-r, HP1/HP2, EHC-U/EHC-L, VAG-F/VAG-R, and ICT37/ICT38) had detection limits of <100 CFU/ml in the presence of gastric tissue. None had 100% specificity or sensitivity; all produced false-positive results. The HP64-f/HP64 r for ureA and HP1/HP2 for 16S rRNA individually had sensitivities and specificities of >90% with gastric biopsy specimens. No combinations of primer pairs improved the results. Using these five primer pairs, 54% of the positive saliva samples were determined to be false positive; both the HP64-f/HP64-r and the HP1/HP2 sets produced false positives with saliva. We conclude that clinicians should not rely on results using current PCR primers alone to decide the H. pylori status of an individual patient or as a basis for treatment decisions. The results of studies based on PCR identification of H. pylori in environmental samples should be viewed with caution. Possibly, specific primers sets can be identified based on the presence of multiple putative virulence factor genes. PMID- 19129408 TI - Evaluation of an automated nucleic acid extractor for hepatitis C virus load quantification. AB - The increasing use of molecular methods strongly motivates clinical laboratories to introduce automated nucleic acid extractors. We compared the easyMAG (bioMerieux) with a manual extraction method for hepatitis C virus (HCV) load quantification (RealTime HCV; Abbott). Both methods were comparable, and, therefore, the easyMAG is suitable to be implemented in our laboratory for the management of HCV-infected patients. PMID- 19129409 TI - Susceptibility of Vibrio parahaemolyticus to tris-dependent DNA degradation during pulsed-field gel electrophoresis. PMID- 19129410 TI - Comparison of viral isolation and multiplex real-time reverse transcription-PCR for confirmation of respiratory syncytial virus and influenza virus detection by antigen immunoassays. AB - We evaluated the Prodesse ProFlu-1 real-time reverse transcription-PCR multiplex assay with the SmartCycler instrument for the detection of human respiratory syncytial virus (RSV) and influenza A and B viruses in comparison to conventional cell culture and antigen immunoassays with the BD Directigen A+B and Binax NOW RSV assays over two successive respiratory virus seasons. Ninety-two percent of the 361 specimens tested were nasopharyngeal aspirates obtained from individual patients, of which 119 were positive for RSV and 59 were positive for influenza virus. The median age of the patients whose specimens were positive for RSV and influenza virus were 6.3 months and 42.4 years, respectively. The specificity of all of the methods tested was >or=99%, and the individual sensitivities of NOW RSV, RSV culture, Directigen A+B, influenza virus culture, and the Proflu-1 PCR for influenza/RSV were 82% (95% confidence interval [CI], 73 to 88), 57% (95% CI, 44 to 69), 59% (95% CI, 44 to 72), 54% (95% CI, 38 to 69), and 98% (95% CI, 93 to 100)/95% (95% CI, 85 to 99), respectively. In a clinical setting where viral isolation is performed to confirm rapid antigen immunoassay results for these common respiratory viruses, one-step real-time reverse transcriptase PCR testing can be a more sensitive and timely confirmatory method. PMID- 19129412 TI - Needle-to-incubator transport time: logistic factors influencing transport time for blood culture specimens. AB - The maximum recommended transport time for blood cultures is 4 h [L. S. Garcia (ed.), 2007 Update: Clinical Microbiology Procedures Handbook, 2nd ed., 2007]. In a previous study, we found that the average transport time was 10 h. In this cohort study, we measured transport times for blood cultures in a larger sample and identified predictors for transport times. A total of 4,322 blood cultures from 1,313 patients were included. The median transport time was 3.5 h, with 47% of cultures exceeding the recommended 4 h. Off-site location and type of clinical specialty were the most important predictors of long transport times. Cultures collected during weekend days or on wards at the largest distances from the laboratory were also associated with long transport times. PMID- 19129411 TI - Development of fully automated determination of marker-specific immunoglobulin G (IgG) avidity based on the avidity competition assay format: application for Abbott Architect cytomegalovirus and Toxo IgG Avidity assays. AB - Determination of the avidity of immunoglobulin G (IgG) directed against a specific marker has become an established diagnostic tool for identifying or excluding acute infections with pathogens. A novel assay format termed AVIcomp (avidity competition based on mass action) circumventing the conventional chaotropic format has been developed for determination of the avidity of marker specific IgG in patient specimens. Its applications for cytomegalovirus (CMV) and Toxoplasma gondii are presented. Specific high-avidity IgG from the patient specimen is selectively blocked using a soluble antigen in a sample pretreatment reagent, and the amount of remaining specific low-avidity IgG is determined relative to that in an untreated control. The comparison of the conventional chaotropic format, represented by the Radim CMV IgG Avidity assay, and the newly developed AVIcomp method, as exemplified by the Architect CMV IgG Avidity assay, on blood drawn within 4 months after seroconversion revealed a sensitivity of 100% (97.3% by an alternative calculation) for the AVIcomp format versus 87.5% (75.7% by an alternative calculation) for the chaotropic avidity assay. The specificity on 312 CMV IgG reactive and CMV IgM nonreactive specimens from pregnant women was 100% for the AVIcomp assay and 99.7% for the conventional avidity assay. The Architect Toxo IgG Avidity assay showed an agreement of 97.2% with the bioMerieux Vidas Toxo IgG Avidity Assay employing chaotropic reagents. These performance data suggest that the AVIcomp format shows superior sensitivity and equivalent specificity for the determination of IgG avidity to assays based on the chaotropic method and that the AVIcomp format may also be applicable to other disease states. PMID- 19129413 TI - Melting curve analysis for rapid detection of topoisomerase gene mutations in Haemophilus influenzae. AB - We established a real-time PCR assay with melting curve analysis to rapidly genotype quinolone resistance-determining regions (QRDRs) of gyrase A and topoisomerase IV genes in Haemophilus influenzae. This assay is a useful tool for the detection of fluoroquinolone resistance and for the early detection of preexisting QRDR mutations. PMID- 19129414 TI - Multicenter evaluation of the Cepheid Xpert methicillin-resistant Staphylococcus aureus (MRSA) test as a rapid screening method for detection of MRSA in nares. AB - The first U.S. multicenter clinical trial to assess the performance of the Cepheid Xpert MRSA assay (Xpert MRSA) was conducted. The assay is a qualitative test designed for the rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) directly from nares swabs. This novel test combines integrated nucleic acid extraction and automated real-time PCR for the detection of a MRSA specific signature sequence. A total of 1,077 nares specimens were collected from seven geographically distinct health care sites across the United States with prevalence rates ranging from 5.2% to 44%. Nares specimens were tested by (i) the Xpert MRSA assay, (ii) direct culture on CHROMagar MRSA medium (direct CM culture), and (iii) broth-enriched culture (Trypticase soy broth with 6.5% sodium chloride) followed by plating onto CHROMagar MRSA medium (broth-enriched CM culture). When direct CM culture was designated the reference method, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA assay were 94.3%, 93.2%, 73.0%, and 98.8%, respectively. When broth-enriched CM culture was used as the reference method, the clinical sensitivity, specificity, PPV, and NPV of the Xpert MRSA assay were 86.3%, 94.9%, 80.5%, and 96.6%, respectively. The BD GeneOhm MRSA (BDGO) assay was performed as a comparative molecular method. No statistical performance differences were observed between the Xpert MRSA and BDGO assays when they were compared to culture methods. From this large-scale, multicenter clinical comparison, we conclude that the Xpert MRSA assay is a simple, rapid, and accurate method for performing active surveillance for MRSA in a variety of health care populations. PMID- 19129415 TI - Phloem loading strategies in three plant species that transport sugar alcohols. AB - Many plants translocate sugar alcohols in the phloem. However, the mechanism(s) of sugar alcohol loading in the minor veins of leaves are debated. We characterized the loading strategies of two species that transport sorbitol (Plantago major and apple [Malus domestica]), and one that transports mannitol (Asarina scandens). Plasmodesmata are abundant at all interfaces in the minor vein phloem of apple, and in one of two types of phloem in the minor veins of A. scandens. Few plasmodesmata are present in the minor veins of P. major. Apple differs from the other two species in that sugar alcohol and sucrose (Suc) are present in much higher concentrations in leaves. Apple leaf tissue exposed to exogenous [(14)C]sorbitol, [(14)C]Suc, or (14)CO(2) did not accumulate radiolabel in the minor veins, as determined by macroautoradiography. P. major minor veins accumulated radiolabel from [(14)C]Suc, [(14)C]sorbitol, and (14)CO(2). A. scandens minor veins accumulated (14)C from [(14)C]Suc and (14)CO(2), but not from [(14)C]mannitol. We conclude that the movement of sugar alcohol from the mesophyll into the phloem in apple and A. scandens is symplastic and passive, but in P. major it involves an apoplastic step and is energized. We also suggest that apple leaves transport sorbitol in high concentrations to avoid the feedback limitation of photosynthesis that would result from driving passive movement of solute into the phloem with high levels of Suc alone. The loading pathways and the mechanisms by which hydrostatic pressure is maintained in the minor vein phloem of these species are discussed. PMID- 19129416 TI - System potentials, a novel electrical long-distance apoplastic signal in plants, induced by wounding. AB - Systemic signaling was investigated in both a dicot (Vicia faba) and a monocot (Hordeum vulgare) plant. Stimuli were applied to one leaf (S-leaf), and apoplastic responses were monitored on a distant leaf (target; T-leaf) with microelectrodes positioned in substomatal cavities of open stomata. Leaves that had been injured by cutting and to which a variety of cations were subsequently added caused voltage transients at the T-leaf, which are neither action potentials nor variation potentials: with respect to the cell interior, the initial polarity of these voltage transients is hyperpolarizing; they do not obey the all-or-none rule but depend on both the concentration and the type of substance added and propagate at 5 to 10 cm min(-1). This response is thought to be due to the stimulation of the plasma membrane H(+)-ATPase, a notion supported by the action of fusicoccin, which also causes such voltage transients to appear on the T-leaf, whereas orthovanadate prevents their propagation. Moreover, apoplastic ion flux analysis reveals that, in contrast to action or variation potentials, all of the investigated ion movements (Ca(2+), K(+), H(+), and Cl(-)) occur after the voltage change begins. We suggest that these wound-induced "system potentials" represent a new type of electrical long-distance signaling in higher plants. PMID- 19129417 TI - Postillumination isoprene emission: in vivo measurements of dimethylallyldiphosphate pool size and isoprene synthase kinetics in aspen leaves. AB - The control of foliar isoprene emission is shared between the activity of isoprene synthase, the terminal enzyme catalyzing isoprene formation from dimethylallyldiphosphate (DMADP), and the pool size of DMADP. Due to limited in vivo information of isoprene synthase kinetic characteristics and DMADP pool sizes, the relative importance of these controls is under debate. In this study, the phenomenon of postillumination isoprene release was employed to develop an in vivo method for estimation of the DMADP pool size and to determine isoprene synthase kinetic characteristics in hybrid aspen (Populus tremula x Populus tremuloides) leaves. The method is based on observations that after switching off the light, isoprene emission continues for 250 to 300 s and that the integral of the postillumination isoprene emission is strongly correlated with the isoprene emission rate before leaf darkening, thus quantitatively estimating the DMADP pool size associated with leaf isoprene emission. In vitro estimates demonstrated that overall leaf DMADP pool was very large, almost an order of magnitude larger than the in vivo pool. Yet, the difference between total DMADP pools in light and in darkness (light-dependent DMADP pool) was tightly correlated with the in vivo estimates of the DMADP pool size that is responsible for isoprene emission. Variation in in vivo DMADP pool size was obtained by varying light intensity and atmospheric CO(2) and O(2) concentrations. From these experiments, the in vivo kinetic constants of isoprene synthase were determined. In vivo isoprene synthase kinetic characteristics suggested that isoprene synthase mainly operates under substrate limitation and that short-term light, CO(2), and O(2) dependencies of isoprene emission result from variation in DMADP pool size rather than from modifications in isoprene synthase activity. PMID- 19129418 TI - The proteome of seed development in the model legume Lotus japonicus. AB - We have characterized the development of seeds in the model legume Lotus japonicus. Like soybean (Glycine max) and pea (Pisum sativum), Lotus develops straight seed pods and each pod contains approximately 20 seeds that reach maturity within 40 days. Histological sections show the characteristic three developmental phases of legume seeds and the presence of embryo, endosperm, and seed coat in desiccated seeds. Furthermore, protein, oil, starch, phytic acid, and ash contents were determined, and this indicates that the composition of mature Lotus seed is more similar to soybean than to pea. In a first attempt to determine the seed proteome, both a two-dimensional polyacrylamide gel electrophoresis approach and a gel-based liquid chromatography-mass spectrometry approach were used. Globulins were analyzed by two-dimensional polyacrylamide gel electrophoresis, and five legumins, LLP1 to LLP5, and two convicilins, LCP1 and LCP2, were identified by matrix-assisted laser desorption ionization quadrupole/time-of-flight mass spectrometry. For two distinct developmental phases, seed filling and desiccation, a gel-based liquid chromatography-mass spectrometry approach was used, and 665 and 181 unique proteins corresponding to gene accession numbers were identified for the two phases, respectively. All of the proteome data, including the experimental data and mass spectrometry spectra peaks, were collected in a database that is available to the scientific community via a Web interface (http://www.cbs.dtu.dk/cgi-bin/lotus/db.cgi). This database establishes the basis for relating physiology, biochemistry, and regulation of seed development in Lotus. Together with a new Web interface (http://bioinfoserver.rsbs.anu.edu.au/utils/PathExpress4legumes/) collecting all protein identifications for Lotus, Medicago, and soybean seed proteomes, this database is a valuable resource for comparative seed proteomics and pathway analysis within and beyond the legume family. PMID- 19129419 TI - Cell culture-induced gradual and frequent epigenetic reprogramming of invertedly repeated tobacco transgene epialleles. AB - Using a two-component transgene system involving two epiallelic variants of the invertedly repeated transgenes in locus 1 (Lo1) and a homologous single-copy transgene locus 2 (Lo2), we have studied the stability of the methylation patterns and trans-silencing interactions in cell culture and regenerated tobacco (Nicotiana tabacum) plants. The posttranscriptionally silenced (PTGS) epiallele of the Lo1 trans-silences and trans-methylates the target Lo2 in a hybrid (Lo1/Lo2 line), while its transcriptionally silenced variant (Lo1E) does not. This pattern was stable over several generations in plants. However, in early Lo1E/Lo2 callus, decreased transgene expression and partial loss of Lo1E promoter methylation compared with leaf tissue in the parental plant were observed. Analysis of small RNA species and coding region methylation suggested that the transgenes were silenced by a PTGS mechanism. The Lo1/Lo2 line remained silenced, but the nonmethylated Lo1 promoter acquired partial methylation in later callus stages. These data indicate that a cell culture process has brought both epialleles to a similar epigenetic ground. Bisulfite sequencing of the 35S promoter within the Lo1 silencer revealed molecules with no, intermediate, and high levels of methylation, demonstrating, to our knowledge for the first time, cell-to-cell methylation diversity of callus. Regenerated plants showed high interindividual but low intraindividual epigenetic variability, indicating that the callus-induced epiallelic variants were transmitted to plants and became fixed. We propose that epigenetic changes associated with dedifferentiation might influence regulatory pathways mediated by trans-PTGS processes. PMID- 19129420 TI - Compromised virus-induced gene silencing in RDR6-deficient plants. AB - RNA silencing in plants serves as a potent antiviral defense mechanism through the action of small interfering RNAs (siRNAs), which direct RNA degradation. siRNAs can be derived directly from the viral genome or via the action of host encoded RNA-dependent RNA polymerases (RDRs). Plant genomes encode multiple RDRs, and it has been demonstrated that plants defective for RDR6 hyperaccumulate several classes of virus. In this study, we compared the effectiveness of virus induced gene silencing (VIGS) and RNA-directed DNA methylation (RdDM) in wild type and RDR6-deficient Nicotiana benthamiana plants. For the potexvirus Potato virus X (PVX) and the potyvirus Plum pox virus (PPV), the efficiency of both VIGS and RdDM were compromised in RDR6-defective plants despite accumulating high levels of viral siRNAs similar to infection of wild-type plants. The reduced efficiency of VIGS and RdDM was unrelated to the size class of siRNA produced and, at least for PVX, was not dependent on the presence of the virus-encoded silencing suppressor protein, 25K. We suggest that primary siRNAs produced from PVX and PPV in the absence of RDR6 may not be good effectors of silencing and that RDR6 is required to produce secondary siRNAs that drive a more effective antiviral response. PMID- 19129421 TI - Constitutive repression and activation of auxin signaling in Arabidopsis. AB - Aux/IAA proteins are proposed to be transcriptional repressors that play a crucial role in auxin signaling by interacting with auxin response factors and repressing early/primary auxin response gene expression. In assays with transfected protoplasts, this repression was previously shown to occur when auxin concentrations in a cell are low, and derepression/activation was observed when auxin concentrations are elevated. Here we show that a stabilized version of the Arabidopsis (Arabidopsis thaliana) IAA17 repressor, when expressed constitutively or in a specific cell type in Arabidopsis plants, confers phenotypes similar to plants with decreased auxin levels. In contrast, a stabilized version of IAA17 that was converted to a transcriptional activator confers phenotypes similar to plants with increased auxin levels, when expressed under the same conditions in Arabidopsis plants. Free auxin levels were unchanged compared to control (DR5:beta-glucuronidase), however, in the seedlings expressing the IAA17 repressor and activator. These results together with our previous results carried out in transfected protoplasts suggest that the hormone auxin can be bypassed to regulate auxin signaling in a cell-autonomous manner in plants. PMID- 19129422 TI - Rapid conversion of persistent atrial fibrillation to sinus rhythm by intravenous AZD7009. AB - This randomized, double-blind trial compared cardioversion rates between AZD7009 infusion (15-minute 3.25 mg/min, 15-minute 4.4 mg/min, or 30-minute 3.25 mg/min) and placebo infusion (15 or 30 minutes) in patients with atrial fibrillation (AF) scheduled for DC cardioversion. One hundred sixty-eight patients were randomized, 167 received study treatment, and 159 were included in perprotocol analyses. The mean duration of current AF episode was 47 days (range, 0.8-92). In the AZD7009 30-minute 3.25 mg/min group, 21 of 42 patients converted within 90 minutes, compared with 7 of 39, 7 of 36, and 0 of 42 patients in the 15-minute 3.25 mg/min, 15-minute 4.4 mg/min, and combined placebo groups, respectively. Patients not converted within 90 minutes underwent DC cardioversion. In patients with AF episodes .05). These results suggest that a potential therapy for doxorubicin-induced cardiotoxicity involves targeting the endogenous cannabinoid system. PMID- 19129424 TI - Poststreptococcal reactive arthritis in children: a retrospective study. AB - The duration of treatment and appropriate guidelines for antibiotic prophylaxis for children with poststreptococcal reactive arthritis (PSRA) have not been determined. The authors performed a retrospective chart review of 40 children with PSRA and examined their clinical features at initial evaluation and at 6, 12, and 24 months. At baseline, 18% (n = 7) had a finding noted on the echocardiogram. Although most patients developed cardiac findings early on in the course of their disease, 2 patients with a normal baseline echocardiogram may have developed findings after 12 months of follow-up. The mean duration of prophylaxis was 22 months. During the follow-up period, there was improvement in Physician's Global Assessment, number of patients with arthralgia, tender and swollen joints, erythrocyte sedimentation rate, anti-streptolysin O, and anti DNAse B antibody titers. The authors conclude that marked improvement in clinical features and laboratory values was seen over time. Patients may benefit with long term cardiac follow-up. PMID- 19129425 TI - Acute pancreatitis--complicating hepatitis E virus infection in a 7-year-old boy with glucose 6 phosphate dehydrogenase deficiency. AB - The association of viral hepatitis and acute pancreatitis is well described in the literature. Most of the cases occur in conjunction with fulminant hepatitis A and hepatitis B virus infections. The recent literature reports increasing number of cases of this complication secondary to hepatitis E virus (HEV) infection, mostly in young adults in regions endemic for the virus. Till date, to the authors' knowledge, there are 14 well-documented cases of HEV-associated acute pancreatitis in the literature. This study reports on a 7-year-old boy from India deficient in glucose 6 phosphate dehydrogenase (G6PD) with moderately severe pancreatitis, manifesting during the course of nonfulminant acute HEV infection. He developed extremely high serum bilirubin levels, probably attributed to the concomitant viral infection and his G6PD status. He recovered completely with conservative therapy. The present child is the youngest ever reported case till date with this complication secondary to HEV infection. PMID- 19129426 TI - An acyl-ghrelin-specific neutralizing antibody inhibits the acute ghrelin mediated orexigenic effects in mice. AB - Ghrelin is a 28-amino acid peptide secreted mainly by the stomach. Acyl-ghrelin, which binds to and activates the growth hormone secretagogue receptor type 1a (GHS-R1a), is considered to be the active form for its orexigenic effects. It has been demonstrated that peripheral administration of ghrelin stimulates food intake and adiposity in rodents and humans. Accordingly, different approaches to antagonize ghrelin/GHS-R1a signaling have been pursued for the treatment of obesity. In the present study, we generated and characterized high-affinity anti acyl ghrelin-specific monoclonal antibodies (mAbs). In vitro, the lead mAb (33A) displayed specific binding to acyl-ghrelin, with an estimated K(d) value < 100 pM. In recombinant receptor cell-based assays, 33A dose-dependently inhibited the ghrelin-mediated calcium signal, with an IC(50) of approximately 3.5 nM. In vivo, ghrelin dose-dependently stimulated food intake in mice, and this effect was fully blocked by a single injection of 33A. In a 4-week chronic study, 33A was shown to effectively bind to endogenous acyl-ghrelin; however, long-term administration of 33A did not affect food intake or body weight gain in a mouse model of diet-induced obesity. Our results indicate that peripheral neutralization of ghrelin can suppress appetite stimulated by a transient surge in ghrelin levels. The lack of long-term effects on body weight control by 33A suggests that compensatory mechanisms may contribute to the regulation of energy balance. PMID- 19129428 TI - Herbal medicines for menopausal symptoms. AB - Many women are now using herbal medicines to try to relieve menopausal symptoms such as hot flushes and night sweats, in light of recent evidence suggesting that hormone replacement therapy (HRT) may increase the likelihood of breast cancer, ovarian cancer, venous thromboembolism, heart attacks and stroke. or example, one survey has suggested that around 40% of women in the UK have used complementary and alternative treatments for their menopausal symptoms.7 Here we review the efficacy and safety of herbal medicines for the relief of such symptoms. PMID- 19129429 TI - Minocycline for acne - an update. AB - Minocycline is an oral tetracycline that, unlike some other drugs in its class, is a once-daily treatment and need not be taken on an empty stomach.1 Such potential advantages together with preferential use in secondary care helped to establish minocycline as the oral tetracycline of choice for acne.2,3 However, concerns over the safety of minocycline and the lack of therapeutic advantage over other tetracyclines have challenged this view.1,4,5 Here we consider how trends in prescribing of minocycline have changed in the UK in recent years. PMID- 19129430 TI - TPMT testing before azathioprine therapy? AB - Azathioprine has been in use for decades as an immunosuppressant treatment for various autoimmune diseases. It is a prodrug of mercaptopurine, a substance that is subsequently metabolised by several alternative pathways, one of which involves the enzyme thiopurine methyltransferase (TPMT). Some people have deficiency of TPMT because of genetic mutations. This has been widely said to occur in around 3 in 1,000 individuals;1 however, studies in recent years have suggested a prevalence of up to 6 in 1,000.2,3 These people are at great risk of developing severe, potentially life-threatening bone marrow toxicity when treated with conventional doses of azathioprine or mercaptopurine. It is possible to test patients for TPMT activity before starting treatment with these drugs. Here we review the evidence about such testing, and discuss whether it should be used for patients being considered for azathioprine therapy. PMID- 19129431 TI - Perceptions of trained leaders on improving the public health impact of three arthritis foundation programs. AB - The Arthritis Foundation (AF) offers effective community-based programs to help manage arthritis, including aquatic, exercise, and self-help programs. Trained leaders can facilitate the adoption, maintenance, and reach of these programs and thus the impact on public health. This study identifies reasons for becoming AF aquatic, exercise, and/or self-help program leaders, AF program reach, and adoption and maintenance challenges encountered by individuals after being trained. Researchers interviewed by telephone 72 participants who attended an AF leader training workshop. Participants reported various reasons for becoming program leaders (e.g., a wish to help others). AF programs were mainly adopted and maintained in urban communities and in fitness/ health clubs, medical centers, or senior centers. Aquatics programs were the most frequently offered, and all programs had low reach (with a mean number of participants of 14.41, 12.50, and 11.00 for aquatic programs, exercise programs, and self-help programs, respectively. Challenges to adopting and maintaining programs include the time of year (e.g., winter, holidays) and lack of a facility to offer the program. PMID- 19129432 TI - Be a Fit Kid: nutrition and physical activity for the fourth grade. AB - This article describes the inclusion of Be a Fit Kid in the fourth-grade curriculum. Be a Fit Kid is a fitness-emphasized physical activity and heart healthy nutrition education program for elementary school children. Five parent education lessons were offered and nutrition workbooks were distributed to parents. Following the 10-week intervention, significant improvements in fitness, body fat, nutrition knowledge, dietary habits, and levels of lipids and lipoproteins were observed in the intervention group compared with baseline levels. Changes in fitness, body fat, and nutrition knowledge were significant compared with the control group. These findings suggest that comprehensive physical activity and nutrition programs included in the school curriculum may be effective for improving cardiovascular health and reducing future risk for lifestyle-related diseases. PMID- 19129433 TI - Developing methods to compare low-education community-based and university-based survey teams. AB - We compared the quality of low-education community-based survey teams to college educated graduate students. Our approach was to develop methods, conduct a pilot survey, and report lessons. Community and university teams conducted surveys from non-overlapping random samples of addresses at a public housing development in Boston, Massachusetts. The two types of teams make a similar number of attempts (122 and 124, respectively), and there was no statistically significant difference between the teams in terms of response rate or amount of missing data. Similarly, there was no statistically significant difference in refusal rate or in responses to questions in the survey. There was, however, evidence that the community teams used data tracking forms improperly. This study suggests that it is possible to study the relative quality of community and university-based teams in terms of data collection. The findings also suggest that the two types of teams may be roughly comparable. PMID- 19129434 TI - Using the chronic care model to address tobacco in health care delivery organizations: a pilot experience in Washington state. AB - This article describes a Washington State-based Systems Change Pilot Project in which the chronic care model and the model for improvement were used as tools to promote tobacco cessation-related changes within a health care system. Three diverse sites participated in the pilot. Site teams tailored plan-do-study-act tests to site circumstances, addressing current resources and barriers to implementing change. Teams tested system changes that incorporated tobacco use documentation into the routine health services provided. Findings from this pilot suggest that (a) even simple changes with minimal disruption of services can make a difference in improving documentation of tobacco use status; (b) changes to routine practices of health organizations may not be sustainable if ongoing quality assurance mechanisms are not developed; and (c) systems implemented for other disease states within the same organization or patient population are not instinctively applied to tobacco, because of a multitude of factors. PMID- 19129435 TI - Delivering culturally sensitive health messages: the process of adapting brochures for grandparents raising grandchildren in Hawai'i. AB - The efficacy of programs to reduce health disparities depends on their ability to deliver messages in a culturally sensitive manner. This article describes the process of designing a series of brochures for grandparents raising grandchildren. National source material on topics important to grandparents (self care, service use, addiction, and grandchildren's difficult behaviors) was put into draft brochures and pilot tested in two focus groups drawn from Native Hawaiian Asian and Pacific Islander populations. Elements of surface and deep levels directed the form and content of the final brochures. On a surface level, these brochures reflect local culture through pictures and language. On a deep level, which integrates cultural beliefs and practices, they reflect the importance of indirect communication and harmonious relationships. The final brochures have been received favorably in the community. The process of adapting educational material with attention to surface and deep levels can serve as a guide for other health promotion materials. PMID- 19129436 TI - Health promotion and illness demotion at prostate cancer support groups. AB - Although health promotion programs can positively influence health practices, men typically react to symptoms, rather than maintain their health, and are more likely to deny than discuss illness-related issues. Prostate cancer support groups (PCSGs) provide an intriguing exception to these practices, in that men routinely discuss ordinarily private illness experiences and engage with self health. This article draws on individual interview data from 52 men, and participant observations conducted at the meetings of 15 groups in British Columbia, Canada to provide insights to how groups simultaneously facilitate health promotion and illness demotion. The study findings reveal how an environment conducive to men's talk was established to normalize prostate cancer and promote the individual and collective health of group members. From a gendered perspective, men both disrupted and embodied dominant ideals of masculinity in how they engaged with their health at PCSGs. PMID- 19129437 TI - WALK Community Grants Scheme: lessons learned in developing and administering a health promotion microgrants program. AB - The Women's Active Living Kits (WALK) Community Grant Scheme was a key component of a federally funded Australian initiative aimed at increasing local capacity to promote and engage priority women's groups in health-related physical activity. Under the program, community groups and organizations were provided with the opportunity to apply and receive small grants to support the development of women's walking groups with the aim of increasing physical activity participation levels in women, supporting innovative community ideas for increasing women's physical activity by improving social structures and environments, or both. This article describes the development and administration of the WALK Community Grant Scheme, outlines challenges and barriers encountered throughout the grant program process, and provides practical insights for replicating this initiative. PMID- 19129438 TI - Herpes simplex virus UL12.5 targets mitochondria through a mitochondrial localization sequence proximal to the N terminus. AB - The herpes simplex virus type 1 (HSV-1) gene UL12 encodes a conserved alkaline DNase with orthologues in all herpesviruses. The HSV-1 UL12 gene gives rise to two separately promoted 3' coterminal mRNAs which encode distinct but related proteins: full-length UL12 and UL12.5, an amino-terminally truncated form that initiates at UL12 codon 127. Full-length UL12 localizes to the nucleus where it promotes the generation of mature viral genomes from larger precursors. In contrast, UL12.5 is predominantly mitochondrial and acts to trigger degradation of the mitochondrial genome early during infection. We examined the basis for these very different subcellular localization patterns. We confirmed an earlier report that the amino-terminal region of full-length UL12 is required for nuclear localization and provide evidence that multiple nuclear localization determinants are present in this region. In addition, we demonstrate that mitochondrial localization of UL12.5 relies largely on sequences located between UL12 residues 185 and 245 (UL12.5 residues 59 to 119). This region contains a sequence that resembles a typical mitochondrial matrix localization signal, and mutations that reduce the positive charge of this element severely impaired mitochondrial localization. Consistent with matrix localization, UL12.5 displayed a detergent extraction profile indistinguishable from that of the matrix protein cyclophilin D. Mitochondrial DNA depletion required the exonuclease activity of UL12.5, consistent with the idea that UL12.5 located within the matrix acts directly to destroy the mitochondrial genome. These results clarify how two highly related viral proteins are targeted to different subcellular locations with distinct functional consequences. PMID- 19129439 TI - Borna disease virus requires cholesterol in both cellular membrane and viral envelope for efficient cell entry. AB - Borna disease virus (BDV), the prototypic member of the family Bornaviridae within the order Mononegavirales, provides an important model for the investigation of viral persistence within the central nervous system (CNS) and of associated brain disorders. BDV is highly neurotropic and enters its target cell via receptor-mediated endocytosis, a process mediated by the virus surface glycoprotein (G), but the cellular factors and pathways determining BDV cell tropism within the CNS remain mostly unknown. Cholesterol has been shown to influence viral infections via its effects on different viral processes, including replication, budding, and cell entry. In this work, we show that cell entry, but not replication and gene expression, of BDV was drastically inhibited by depletion of cellular cholesterol levels. BDV G-mediated attachment to BDV susceptible cells was cholesterol independent, but G localized to lipid rafts (LR) at the plasma membrane. LR structure and function critically depend on cholesterol, and hence, compromised structural integrity and function of LR caused by cholesterol depletion likely inhibited the initial stages of BDV cell internalization. Furthermore, we also show that viral-envelope cholesterol is required for BDV infectivity. PMID- 19129440 TI - Partial protection of Simian immunodeficiency virus (SIV)-infected rhesus monkeys against superinfection with a heterologous SIV isolate. AB - Although there is increasing evidence that individuals already infected with human immunodeficiency virus type 1 (HIV-1) can be infected with a heterologous strain of the virus, the extent of protection against superinfection conferred by the first infection and the biologic consequences of superinfection are not well understood. We explored these questions in the simian immunodeficiency virus (SIV)/rhesus monkey model of HIV-1/AIDS. We infected cohorts of rhesus monkeys with either SIVmac251 or SIVsmE660 and then exposed animals to the reciprocal virus through intrarectal inoculations. Employing a quantitative real-time PCR assay, we determined the replication kinetics of the two strains of virus for 20 weeks. We found that primary infection with a replication-competent virus did not protect against acquisition of infection by a heterologous virus but did confer relative control of the superinfecting virus. In animals that became superinfected, there was a reduction in peak replication and rapid control of the second virus. The relative susceptibility to superinfection was not correlated with CD4(+) T-cell count, CD4(+) memory T-cell subsets, cytokine production by virus-specific CD8(+) or CD4(+) cells, or neutralizing antibodies at the time of exposure to the second virus. Although there were transient increases in viral loads of the primary virus and a modest decline in CD4(+) T-cell counts after superinfection, there was no evidence of disease acceleration. These findings indicate that an immunodeficiency virus infection confers partial protection against a second immunodeficiency virus infection, but this protection may be mediated by mechanisms other than classical adaptive immune responses. PMID- 19129441 TI - Identifying sites bound by Epstein-Barr virus nuclear antigen 1 (EBNA1) in the human genome: defining a position-weighted matrix to predict sites bound by EBNA1 in viral genomes. AB - We identified binding sites for Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) in the human genome using chromatin immunoprecipitation and microarrays. The sequences for these newly identified sites were used to generate a position weighted matrix (PWM) for EBNA1's DNA-binding sites. This PWM helped identify additional DNA-binding sites for EBNA1 in the genomes of EBV, Kaposi's sarcoma associated herpesvirus, and cercopithecine herpesvirus 15 (CeHV-15) (also called herpesvirus papio 15). In particular, a homologue of the Rep* locus in EBV was predicted in the genome of CeHV-15, which is notable because Rep* of EBV was not predicted by the previously developed consensus sequence for EBNA1's binding DNA. The Rep* of CeHV-15 functions as an origin of DNA synthesis in the EBV-positive cell line Raji; this finding thus builds on a set of DNA-binding sites for EBNA1 predicted in silico. PMID- 19129442 TI - Inhibition of the ubiquitin-proteasome system prevents vaccinia virus DNA replication and expression of intermediate and late genes. AB - The ubiquitin-proteasome system has a central role in the degradation of intracellular proteins and regulates a variety of functions. Viruses belonging to several different families utilize or modulate the system for their advantage. Here we showed that the proteasome inhibitors MG132 and epoxomicin blocked a postentry step in vaccinia virus (VACV) replication. When proteasome inhibitors were added after virus attachment, early gene expression was prolonged and the expression of intermediate and late genes was almost undetectable. By varying the time of the removal and addition of MG132, the adverse effect of the proteasome inhibitors was narrowly focused on events occurring 2 to 4 h after infection, the time of the onset of viral DNA synthesis. Further analyses confirmed that genome replication was inhibited by both MG132 and epoxomicin, which would account for the effect on intermediate and late gene expression. The virus-induced replication of a transfected plasmid was also inhibited, indicating that the block was not at the step of viral DNA uncoating. UBEI-41, an inhibitor of the ubiquitin-activating enzyme E1, also prevented late gene expression, supporting the role of the ubiquitin-proteasome system in VACV replication. Neither the overexpression of ubiquitin nor the addition of an autophagy inhibitor was able to counter the inhibitory effects of MG132. Further studies of the role of the ubiquitin-proteasome system for VACV replication may provide new insights into virus-host interactions and suggest potential antipoxviral drugs. PMID- 19129444 TI - Evolution of the Sabin vaccine into pathogenic derivatives without appreciable changes in antigenic properties: need for improvement of current poliovirus surveillance. AB - The Sabin oral polio vaccine (OPV) may evolve into pathogenic viruses, causing sporadic cases and outbreaks of poliomyelitis. Such vaccine-derived polioviruses (VDPV) generally exhibit altered antigenicity. The current paradigm to distinguish VDPV from OPV and wild polioviruses is to characterize primarily those poliovirus isolates that demonstrate deviations from OPV in antigenic and genetic intratypic differentiation (ITD) tests. Here we report on two independent cases of poliomyelitis caused by VDPVs with "Sabin-like" properties in several ITD assays. The results suggest the existence of diverse pathways of OPV evolution and necessitate improvement of poliovirus surveillance, which currently potentially misses this class of VDPV. PMID- 19129443 TI - A basic patch on alpha-adaptin is required for binding of human immunodeficiency virus type 1 Nef and cooperative assembly of a CD4-Nef-AP-2 complex. AB - A critical function of the human immunodeficiency virus type 1 Nef protein is the downregulation of CD4 from the surfaces of infected cells. Nef is believed to act by linking the cytosolic tail of CD4 to the endocytic machinery, thereby increasing the rate of CD4 internalization. In support of this model, weak binary interactions between CD4, Nef, and the endocytic adaptor complex, AP-2, have been reported. In particular, dileucine and diacidic motifs in the C-terminal flexible loop of Nef have been shown to mediate binding to a combination of the alpha and sigma2 subunits of AP-2. Here, we report the identification of a potential binding site for the Nef diacidic motif on alpha-adaptin. This site comprises two basic residues, lysine-297 and arginine-340, on the alpha-adaptin trunk domain. The mutation of these residues specifically inhibits the ability of Nef to bind AP-2 and downregulate CD4. We also present evidence that the diacidic motif on Nef and the basic patch on alpha-adaptin are both required for the cooperative assembly of a CD4-Nef-AP-2 complex. This cooperativity explains how Nef is able to efficiently downregulate CD4 despite weak binary interactions between components of the tripartite complex. PMID- 19129445 TI - Sindbis virus-based measles DNA vaccines protect cotton rats against respiratory measles: relevance of antibodies, mucosal and systemic antibody-secreting cells, memory B cells, and Th1-type cytokines as correlates of immunity. AB - Measles remains an important cause of pediatric morbidity and mortality in developing countries, especially among infants who are too young to receive the current licensed live attenuated measles vaccine. We developed two Sindbis virus DNA vaccines encoding the measles virus hemagglutinin (pMSIN-H) and fusion proteins (pMSINH-FdU) and examined their immunogenicities and protective efficacies when administered alone or followed by the live measles virus vaccine in cotton rats. Neutralizing antibodies, mucosal and systemic antibody-secreting cells, memory B cells, and gamma interferon-secreting T cells developed after priming and increased after boosting. pMSIN-H priming conferred 100% protection against pulmonary measles, whereas pMSINH-FdU protected only in conjunction with the live measles virus vaccine boost. PMID- 19129446 TI - Generation of herpesvirus entry mediator (HVEM)-restricted herpes simplex virus type 1 mutant viruses: resistance of HVEM-expressing cells and identification of mutations that rescue nectin-1 recognition. AB - Both initial infection and cell-to-cell spread by herpes simplex virus type 1 (HSV-1) require the interaction of the viral glycoprotein D (gD) with an entry receptor on the cell surface. The two major HSV entry receptors, herpesvirus entry mediator (HVEM) and nectin-1, mediate infection independently but are coexpressed on a variety of cells. To determine if both receptors are active in these instances, we have established mutant viruses that are selectively impaired for recognition of one or the other receptor. In plaque assays, these viruses showed approximately 1,000-fold selectivity for the matched receptor over the mismatched receptor. Separate assays showed that each virus is impaired for both infection and spread through the mismatched receptor. We tested several human tumor cell lines for susceptibility to these viruses and observed that HT29 colon carcinoma cells are susceptible to infection by nectin-1-restricted virus but are highly resistant to HVEM-restricted virus infection, despite readily detectable HVEM expression on the cell surface. HVEM cDNA isolated from HT29 cells rendered HSV-resistant cells permissive for infection by the HVEM-restricted virus, suggesting that HT29 cells lack a cofactor for HVEM-mediated infection or express an HVEM-specific inhibitory factor. Passaging of HVEM-restricted virus on nectin 1-expressing cells yielded a set of gD missense mutations that each restored functional recognition of nectin-1. These mutations identify residues that likely play a role in shaping the nectin-1 binding site of gD. Our findings illustrate the utility of these receptor-restricted viruses in studying the early events in HSV infection. PMID- 19129447 TI - Modified vaccinia virus ankara triggers chemotaxis of monocytes and early respiratory immigration of leukocytes by induction of CCL2 expression. AB - Orthopoxviruses commonly enter into humans and animals via the respiratory tract. Herein, we show that immigration of leukocytes into the lung is triggered via intranasal infection of mice with modified vaccinia virus Ankara (MVA) and not with the vaccinia virus (VACV) Elstree, Wyeth, or Western Reserve (WR) strain. Immigrating cells were identified as monocytes, neutrophils, and CD4(+) lymphocytes by flow cytometry and could be detected 24 h and 48 h postinfection. Using an in vitro chemotaxis assay, we confirmed that infection with MVA induces the expression of a soluble chemotactic factor for monocytes, identified as CCL2 (monocyte chemotactic protein-1 [MCP-1]). In contrast to infection with several other VACV strains, MVA induced the expression of CCL2, CCL3, CCL4, and CXCL10 in the human monocytic cell line THP-1 as well as in primary human monocytes. Thus, MVA, and not the VACV Elstree, Wyeth, or WR strain, consistently triggered the expression of a panel of chemokines, including CCL2, in the murine lung, correlating considerably with the immigration of leukocytes. Using CCL2-deficient mice, we demonstrate that CCL2 plays a key role in MVA-triggered respiratory immigration of leukocytes. Moreover, UV irradiation of MVA prevented CCL2 expression in vitro and in vivo as well as respiratory immigration of leukocytes, demonstrating the requirement for an activated molecular viral life cycle. We propose that MVA-triggered chemokine expression causes early immigration of leukocytes to the site of infection, a feature that is important for rapid immunization and its safety and efficiency as a viral vector. PMID- 19129449 TI - An HLA-A2-restricted T-cell epitope mapped to the BNLF2a immune evasion protein of Epstein-Barr virus that inhibits TAP. AB - The early lytic cycle protein of Epstein-Barr virus (EBV), BNLF2a, has recently been shown to play a critical role in immune evasion by inhibiting the peptide transporter associated with antigen processing (TAP), thereby blocking antigen specific CD8(+) T-cell recognition of many lytic cycle antigens. Surprisingly, we now show that a peptide ((50)VLFGLLCLL(58)) from the hydrophobic C-terminal region of this small (60-amino-acid) EBV protein is efficiently presented by the common class I allele HLA-A2 for recognition by cytotoxic T lymphocytes. The mechanism for this unexpected finding was revealed by experiments showing that this epitope is processed and presented independently of TAP. PMID- 19129448 TI - High specific infectivity of plasma virus from the pre-ramp-up and ramp-up stages of acute simian immunodeficiency virus infection. AB - To define the ratio of simian immunodeficiency virus (SIV) RNA molecules to infectious virions in plasma, a ramp-up-stage plasma pool was made from the earliest viral RNA (vRNA)-positive plasma samples (collected approximately 7 days after inoculation) from seven macaques, and a set-point-stage plasma pool was made from plasma samples collected 10 to 16 weeks after peak viremia from seven macaques; vRNA levels in these plasma pools were determined, and serial 10-fold dilutions containing 1 to 1,500 vRNA copies/ml were made. Intravenous (i.v.) inoculation of a 1-ml aliquot of diluted ramp-up-stage plasma containing 20 vRNA copies infected 2 of 2 rhesus macaques, while for the set-point-stage plasma, i.v. inoculation with 1,500 vRNA copies was needed to transmit infection. Further, when the heat-inactivated set-point-stage plasma pool was mixed with ramp-up-stage virions, infection of inoculated macaques was blocked. Notably, 2 of 2 animals inoculated with 85 ml of a pre-ramp-up plasma pool containing <3 SIV RNA copies/ml developed SIV infections characterized by high levels of viral replication, demonstrating that "vRNA-negative" plasma collected from macaques in the pre-ramp-up stage is infectious. Furthermore, there is a high ratio of infectious virions to total virions in ramp-up-stage plasma (between 1:1 and 1:10) and a lower ratio in set-point-stage plasma (between 1:75 and 1:750). Heat inactivated chronic-stage plasma can "neutralize" the highly infectious ramp-up stage virions. These findings have implications for the understanding of the natural history of SIV and human immunodeficiency virus infection and transmission. PMID- 19129450 TI - Internal initiation stimulates production of p8 minicore, a member of a newly discovered family of hepatitis C virus core protein isoforms. AB - The hepatitis C virus (HCV) core gene is more conserved at the nucleic acid level than is necessary to preserve the sequence of the core protein, suggesting that it contains information for additional functions. We used a battery of anticore antibodies to test the hypothesis that the core gene directs the synthesis of core protein isoforms. Infectious viruses, replicons, and RNA transcripts expressed a p8 minicore containing the C-terminal portion of the p21 core protein and lacking the N-terminal portion. An interferon resistance mutation, U271A, which creates an AUG at codon 91, upregulated p8 expression in Con1 replicons, suggesting that p8 is produced by an internal initiation event and that 91-AUG is the preferred, but not the required, initiation codon. Synthesis of p8 was independent of p21, as shown by the abundant production of p8 from transcripts containing an UAG stop codon that blocked p21 production. Three infectious viruses, JFH-1 (2a core), J6/JFH (2a core), and H77/JFH (1a core), and a bicistronic construct, Bi-H77/JFH, all expressed both p8 and larger isoforms. The family of minicores ranges in size from 8 to 14 kDa. All lack the N-terminal portion of the p21 core. In conclusion, the core gene contains an internal signal that stimulates the initiation of protein synthesis at or near codon 91, leading to the production of p8. Infectious viruses of both genotype 1 and 2 HCV express a family of larger isoforms, in addition to p8. Minicores lack significant portions of the RNA binding domain of p21 core. Studies are under way to determine their functions. PMID- 19129451 TI - Reovirus FAST protein transmembrane domains function in a modular, primary sequence-independent manner to mediate cell-cell membrane fusion. AB - The FAST proteins are a unique family of virus-encoded cell-cell membrane fusion proteins. In the absence of a cleavable N-terminal signal peptide, a single-pass transmembrane domain (TMD) functions as a reverse signal-anchor to direct the FAST proteins into the plasma membrane in an N(exo)/C(cyt) topology. There is little information available on the role of the FAST protein TMD in the cell-cell membrane fusion reaction. We show that in the absence of conservation in the length or primary amino acid sequence, the p14 TMD can be functionally exchanged with the TMDs of the p10 and p15 FAST proteins. This is not the case for chimeric p14 proteins containing the TMDs of two different enveloped viral fusion proteins or a cellular membrane protein; such chimeric proteins were defective for both pore formation and syncytiogenesis. TMD structural features that are conserved within members of the FAST protein family presumably play direct roles in the fusion reaction. Molecular modeling suggests that the funnel-shaped architecture of the FAST protein TMDs may represent such a conserved structural and functional motif. Interestingly, although heterologous TMDs exert diverse influences on the trafficking of the p14 FAST protein, these TMDs are capable of functioning as reverse signal-anchor sequences to direct p14 into lipid rafts in the correct membrane topology. The FAST protein TMDs are therefore not primary determinants of type III protein topology, but they do play a direct, sequence-independent role in the membrane fusion reaction. PMID- 19129452 TI - E4orf1 limits the oncolytic potential of the E1B-55K deletion mutant adenovirus. AB - Clinical trials have shown oncolytic adenoviruses to be tumor selective with minimal toxicity toward normal tissue. The virus ONYX-015, in which the gene encoding the early region 1B 55-kDa (E1B-55K) protein is deleted, has been most effective when used in combination with either chemotherapy or radiation therapy. Therefore, improving the oncolytic nature of tumor-selective adenoviruses remains an important objective for improving this form of cancer therapy. Cells infected during the G(1) phase of the cell cycle with the E1B-55K deletion mutant virus exhibit a reduced rate of viral late protein synthesis, produce fewer viral progeny, and are less efficiently killed than cells infected during the S phase. Here we demonstrate that the G(1) restriction imposed on the E1B-55K deletion mutant virus is due to the viral oncogene encoded by open reading frame 1 of early region 4 (E4orf1). E4orf1 has been reported to signal through the phosphatidylinositol 3'-kinase pathway leading to the activation of Akt, mTOR, and p70 S6K. Evidence presented here shows that E4orf1 may also induce the phosphorylation of Akt and p70 S6K in a manner that depends on Rac1 and its guanine nucleotide exchange factor Tiam1. Accordingly, agents that have been reported to disrupt the Tiam1-Rac1 interaction or to prevent phosphorylation of the ribosomal S6 kinase partially alleviated the E4orf1 restriction to late viral protein synthesis and enhanced tumor cell killing by the E1B-55K mutant virus. These results demonstrate that E4orf1 limits the oncolytic nature of a conditionally replicating adenovirus such as ONYX-015. The therapeutic value of similar oncolytic adenoviruses may be improved by abrogating E4orf1 function. PMID- 19129453 TI - Mitogen-activated protein kinase-activated kinase RSK2 plays a role in innate immune responses to influenza virus infection. AB - Viral infections induce signaling pathways in mammalian cells that stimulate innate immune responses and affect cellular processes, such as apoptosis, mitosis, and differentiation. Here, we report that the ribosomal protein S6 kinase alpha 3 (RSK2), which is activated through the "classical" mitogen activated protein kinase pathway, plays a role in innate immune responses to influenza virus infection. RSK2 functions in the regulation of cell growth and differentiation but was not known to play a role in the cellular antiviral response. We have found that knockdown of RSK2 enhanced viral polymerase activity and growth of influenza viruses. Influenza virus infection stimulates NK-kappaB- and beta interferon-dependent promoters. This stimulation was reduced in RSK2 knockdown cells, suggesting that RSK2 executes its effect through innate immune response pathways. Furthermore, RSK2 knockdown suppressed influenza virus-induced phosphorylation of the double-stranded RNA-activated protein kinase PKR, a known antiviral protein. These findings establish a role for RSK2 in the cellular antiviral response. PMID- 19129454 TI - P64, a novel major virion DNA-binding protein potentially involved in condensing the Spodoptera frugiperda Ascovirus 1a genome. AB - We recently identified 21 structural proteins in the virion of Spodoptera frugiperda ascovirus 1a (SfAV1a), a virus with a large, double-stranded DNA genome of 157 kbp, which attacks species of the lepidopteran family Noctuidae. The two most abundant virion proteins were the major capsid protein and a novel protein (P64) of 64 kDa that contained two distinct domains not known previously to occur together. The amino-terminal half of P64 (residues 1 to 263) contained four repeats (a recently recognized motif with an unknown function) of a virus specific two-cysteine adaptor. Adjoined to this, the carboxy-terminal half of P64 (residues 279 to 455) contained 14 copies of a highly basic, tandemly repeated motif rich in arginine and serine, having an 11- to 13-amino-acid consensus sequence, SPSQRRSTS(V/K)(A/S)RR, yielding a predicted isoelectric point of 12.2 for this protein. In the present study, we demonstrate by Southwestern analysis that SfAV1a P64 was the only virion structural protein that bound DNA. Additional electrophoretic mobility shift assays showed that P64 bound SfAV1a as well as non SfAV1a DNA. Furthermore, we show through immunogold labeling of ultrathin sections that P64 is a component of virogenic stroma and appears to be progressively incorporated into the SfAV1a DNA core during virion assembly. As no other virion structural protein bound DNA and no basic DNA-binding proteins of lower mass are encoded by the SfAV1a genome or were identified by proteomic analysis, our results suggest that P64's function is to condense the large genome of this virus and assist in packaging this genome into its virion. PMID- 19129455 TI - Differentially stimulated CD4+ T cells display altered human immunodeficiency virus infection kinetics: implications for the efficacy of antiviral agents. AB - The activation state of CD4(+) T cells plays a crucial role in the establishment of a productive human immunodeficiency virus infection. Here, we show that T cells stimulated for 1 day demonstrated delayed kinetics of viral reverse transcription and integration compared to cells stimulated for 2 days prior to infection. As a result, the efficiency of reverse transcription and integration inhibitors differs in these differentially stimulated cells. These studies increase our understanding of how T cells support viral replication and provide insight regarding the efficiency of antiretroviral therapy in lymphoid compartments. PMID- 19129456 TI - Abrogation of the postmitotic checkpoint contributes to polyploidization in human papillomavirus E7-expressing cells. AB - High-risk types of human papillomavirus (HPV) are considered the major causative agents of cervical carcinoma. The transforming ability of HPV resides in the E6 and E7 oncogenes, yet the pathway to transformation is not well understood. Cells expressing the oncogene E7 from high-risk HPVs have a high incidence of polyploidy, which has been shown to occur as an early event in cervical carcinogenesis and predisposes the cells to aneuploidy. The mechanism through which E7 contributes to polyploidy is not known. It has been hypothesized that E7 induces polyploidy in response to mitotic stress by abrogating the mitotic spindle assembly checkpoint. It was also proposed that E7 may stimulate rereplication to induce polyploidy. We have tested these hypotheses by using human epithelial cells in which E7 expression induces a significant amount of polyploidy. We find that E7-expressing cells undergo normal mitoses with an intact spindle assembly checkpoint and that they are able to complete cytokinesis. Our results also exclude DNA rereplication as a major mechanism of polyploidization in E7-expressing cells upon microtubule disruption. Instead, we have shown that while normal cells arrest at the postmitotic checkpoint after adaptation to the spindle assembly checkpoint, E7-expressing cells replicate their DNA and propagate as polyploid cells. Thus, abrogation of the postmitotic checkpoint leads to polyploidy formation in E7-expressing human epithelial cells. Our results suggest that downregulation of pRb is important for E7 to induce polyploidy and abrogation of the postmitotic checkpoint. PMID- 19129457 TI - Determinants flanking the CD4 binding loop modulate macrophage tropism of human immunodeficiency virus type 1 R5 envelopes. AB - Human immunodeficiency virus type 1 R5 viruses vary extensively in phenotype. Thus, R5 envelopes (env) in the brain tissue of individuals with neurological complications are frequently highly macrophage-tropic. Macrophage tropism correlates with the capacity of the envelope to exploit low CD4 levels for infection. In addition, the presence of an asparagine at residue 283 within the CD4 binding site has been associated with brain-derived envelopes, increased env CD4 affinity, and enhanced macrophage tropism. Here, we identify additional envelope determinants of R5 macrophage tropism. We compared highly macrophage tropic (B33) and non-macrophage-tropic (LN40) envelopes from brain and lymph node specimens of one individual. We first examined the role of residue 283 in macrophage tropism. Introduction of N283 into LN40 (T283N) conferred efficient macrophage infectivity. In contrast, substitution of N283 for the more conserved threonine in B33 had little effect on macrophage infection. Thus, B33 carried determinants for macrophage tropism that were independent of N283. We prepared chimeric B33/LN40 envelopes and used site-directed mutagenesis to identify additional determinants. The determinants of macrophage tropism that were identified included residues on the CD4 binding loop flanks that were proximal to CD4 contact residues and residues in the V3 loop. The same residues affected sensitivity to CD4-immunoglobulin G inhibition, consistent with an altered env CD4 affinity. We predict that these determinants alter exposure of CD4 contact residues. Moreover, the CD4 binding loop flanks are variable and may contribute to a general mechanism for protecting proximal CD4 contact residues from neutralizing antibodies. Our results have relevance for env-based vaccines that will need to expose critical CD4 contact residues to the immune system. PMID- 19129458 TI - A systems biology approach to identify the combination effects of human herpesvirus 8 genes on NF-kappaB activation. AB - Human herpesvirus 8 (HHV-8) is the etiologic agent of Kaposi's sarcoma and primary effusion lymphoma. Activation of the cellular transcription factor nuclear factor-kappa B (NF-kappaB) is essential for latent persistence of HHV-8, survival of HHV-8-infected cells, and disease progression. We used reverse transfected cell microarrays (RTCM) as an unbiased systems biology approach to systematically analyze the effects of HHV-8 genes on the NF-kappaB signaling pathway. All HHV-8 genes individually (n = 86) and, additionally, all K and latent genes in pairwise combinations (n = 231) were investigated. Statistical analyses of more than 14,000 transfections identified ORF75 as a novel and confirmed K13 as a known HHV-8 activator of NF-kappaB. K13 and ORF75 showed cooperative NF-kappaB activation. Small interfering RNA-mediated knockdown of ORF75 expression demonstrated that this gene contributes significantly to NF kappaB activation in HHV-8-infected cells. Furthermore, our approach confirmed K10.5 as an NF-kappaB inhibitor and newly identified K1 as an inhibitor of both K13- and ORF75-mediated NF-kappaB activation. All results obtained with RTCM were confirmed with classical transfection experiments. Our work describes the first successful application of RTCM for the systematic analysis of pathofunctions of genes of an infectious agent. With this approach, ORF75 and K1 were identified as novel HHV-8 regulatory molecules on the NF-kappaB signal transduction pathway. The genes identified may be involved in fine-tuning of the balance between latency and lytic replication, since this depends critically on the state of NF kappaB activity. PMID- 19129459 TI - Comparisons of CD8+ T cells specific for human immunodeficiency virus, hepatitis C virus, and cytomegalovirus reveal differences in frequency, immunodominance, phenotype, and interleukin-2 responsiveness. AB - To better understand the components of an effective immune response to human immunodeficiency virus (HIV), the CD8(+) T-cell responses to HIV, hepatitis C virus (HCV), and cytomegalovirus (CMV) were compared with regard to frequency, immunodominance, phenotype, and interleukin-2 (IL-2) responsiveness. Responses were examined in rare patients exhibiting durable immune-mediated control over HIV, termed long-term nonprogressors (LTNP) or elite controllers, and patients with progressive HIV infection (progressors). The magnitude of the virus-specific CD8(+) T-cell response targeting HIV, CMV, and HCV was not significantly different between LTNP and progressors, even though their capacity to proliferate to HIV antigens was preserved only in LTNP. In contrast to HIV-specific CD8(+) T cell responses of LTNP, HLA B5701-restricted responses within CMV pp65 were rare and did not dominate the total CMV-specific response. Virus-specific CD8(+) T cells were predominantly CD27(+)45RO(+) for HIV and CD27(-)45RA(+) for CMV; however, these phenotypes were highly variable and heavily influenced by the degree of viremia. Although IL-2 induced significant expansions of CMV-specific CD8(+) T cells in LTNP and progressors by increasing both the numbers of cells entering the proliferating pool and the number of divisions, the proliferative capacity of a significant proportion of HIV-specific CD8(+) T cells was not restored with exogenous IL-2. These results suggest that immunodominance by HLA B5701-restricted cells is specific to HIV infection in LTNP and is not a feature of responses to other chronic viral infections. They also suggest that poor responsiveness to IL-2 is a property of HIV-specific CD8(+) T cells of progressors that is not shared with responses to other viruses over which immunologic control is maintained. PMID- 19129460 TI - Papillomavirus E2 proteins and the host BRD4 protein associate with transcriptionally active cellular chromatin. AB - The interaction of papillomavirus E2 proteins with cellular Brd4 protein is important for transcriptional regulation of viral genes and partitioning of viral genomes. Bovine papillomavirus type 1 (BPV-1) E2 binds cellular chromatin in complex with Brd4 in both mitotic and interphase cells. To identify specific sites of E2 interaction on cellular chromatin, a genome-wide chromatin immunoprecipitation-on-chip analysis was carried out using human promoter sequences. Both E2 and Brd4 were found bound to most transcriptionally active promoters in C33A cells. These promoters were also bound by RNA polymerase II and were modified by histone H3 acetylation and K4 trimethylation, all indicators of active transcription. E2 binding strongly correlated with Brd4 and RNA polymerase II occupancy and H3K4me3 modification at all human promoters, indicating that E2 bound to active promoters. E2 binding did not correlate with the presence of consensus E2 binding sites in the promoters. Furthermore, the mRNA levels of E2 bound cellular genes were not significantly changed by E2 expression. Thus, the papillomavirus E2 proteins bind to transcriptionally active cellular genes but do not change their activity. We propose that this may be a way for the virus to ensure that the viral genome is retained in transcriptionally active regions of the nucleus to escape silencing. Therefore, E2-mediated tethering of viral genomes to host chromatin has multiple roles: to partition the viral genome to daughter cells, to ensure that the genomes are retained in the nucleus, and to make certain that the genomes are retained in functionally active nuclear domains. PMID- 19129461 TI - REDD2 is enriched in skeletal muscle and inhibits mTOR signaling in response to leucine and stretch. AB - The protein kinase mammalian target of rapamycin (mTOR) is well established as a key regulator of skeletal muscle size. In this study, we determined that the stress responsive gene REDD2 (regulated in development and DNA damage responses 2) is a negative regulator of mTOR signaling and is expressed predominantly in skeletal muscle. Overexpression of REDD2 in muscle cells significantly inhibited basal mTOR signaling and diminished the response of mTOR to leucine addition or mechanical stretch. The inhibitory function of REDD2 on mTOR signaling seems to be mediated downstream or independent of Akt signaling and upstream of Rheb (Ras homolog enriched in brain). Knock down of tuberous sclerosis complex 2 (TSC2) using small interfering (si)RNA potently activated mTOR signaling and was sufficient to rescue REDD2 inhibition of mTOR activity, suggesting that REDD2 functions by modulating TSC2 function. Immunoprecipitation assays demonstrated that REDD2 does not directly interact with either TSC1 or TSC2. However, we found that REDD2 forms a complex with 14-3-3 protein and that increasing expression of REDD2 acts to competitively dissociate TSC2 from 14-3-3 and inhibits mTOR signaling. These findings demonstrate that REDD2 is a skeletal muscle specific inhibitory modulator of mTOR signaling and identify TSC2 and 14-3-3 as key molecular links between REDD2 and mTOR function. PMID- 19129462 TI - Regulation of connexin gene expression during skeletal muscle regeneration in the adult rat. AB - In the adult skeletal muscle, various kinds of trauma promote proliferation of satellite cells that differentiate into myoblasts forming new myofibers or to repair the damaged one. The aim of present work was to perform a comparative spatial and temporal analysis of connexin (Cx) 37, Cx39, Cx40, Cx43, and Cx45 expression in the adult regenerating skeletal muscle in response to crush injury. Within 24 h from injury, Cx37 expression was upregulated in the endothelial cells of blood vessels, and, 5 days after injury, Cx37-expressing cells were found inside the area of lesion and formed clusters generating new blood vessels with endothelial cells expressing Cx37. Three days after injury, Cx39 mRNA was selectively expressed in myogenin-positive cells, forming rows of closely apposed cell nuclei fusing in myotubes. Cx40 mRNA-labeled cells were observed within 24 h from injury in the endothelium of blood vessels, and, 5 days after lesion, Cx40 labeled cells were found inside the area of lesion-forming rows of myogenin positive, closely apposed cells coexpressing Cx39. Within 24 h from lesion, both Cx43 and Cx45 mRNAs were upregulated in individual cells, and some of them were positive for M-cadherin. Three days after injury, a large number of both Cx43 and Cx45 mRNA-labeled and myogenin-positive cells were found inside the area of lesion. Taken together, these results show that at least four Cxs, out of five expressed in regenerating skeletal muscle, can be differentially involved in communication of myogenic cells during the process of cell proliferation, aggregation, and fusion to form new myotubes or to repair damaged myofibers. PMID- 19129463 TI - Mechanisms of pH-gradient driven transport mediated by organic anion polypeptide transporters. AB - Organic anion transporting polypeptides (humans OATPs, rodents Oatps) are expressed in most mammalian tissues and mediate cellular uptake of a wide variety of amphipathic organic compounds such as bile salts, steroid conjugates, oligopeptides, and a large list of drugs, probably by acting as anion exchangers. In the present study we aimed to investigate the role of the extracellular pH on the transport activity of nine human and four rat OATPs/Oatps. Furthermore, we aimed to test the concept that OATP/Oatp transport activity is accompanied by extrusion of bicarbonate. By using amphibian Xenopus laevis oocytes expressing OATPs/Oatps and mammalian cell lines stably transfected with OATPs/Oatps, we could demonstrate that in all OATPs/Oatps investigated, with the exception of OATP1C1, a low extracellular pH stimulated transport activity. This stimulation was accompanied by an increased substrate affinity as evidenced by lower apparent Michaelis-Menten constant values. OATP1C1 is lacking a highly conserved histidine in the third transmembrane domain, which was shown by site-directed mutagenesis to be critically involved in the pH dependency of OATPs/Oatps. Using online intracellular pH measurements in OATP/Oatp-transfected Chinese Hamster Ovary (CHO)-K1 cells, we could demonstrate the presence of a 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid-sensitive chloride/bicarbonate exchanger in CHO-K1 cells and that OATP/Oatp-mediated substrate transport is paralleled by bicarbonate efflux. We conclude that the pH dependency of OATPs/Oatps may lead to a stimulation of substrate transport in an acidic microenvironment and that the OATP/Oatp-mediated substrate transport into cells is generally compensated or accompanied by bicarbonate efflux. PMID- 19129464 TI - Myostatin regulates fiber-type composition of skeletal muscle by regulating MEF2 and MyoD gene expression. AB - Myostatin (Mstn) is a secreted growth factor belonging to the tranforming growth factor (TGF)-beta superfamily. Inactivation of murine Mstn by gene targeting, or natural mutation of bovine or human Mstn, induces the double muscling (DM) phenotype. In DM cattle, Mstn deficiency increases fast glycolytic (type IIB) fiber formation in the biceps femoris (BF) muscle. Using Mstn null ((-/-)) mice, we suggest a possible mechanism behind Mstn-mediated fiber-type diversity. Histological analysis revealed increased type IIB fibers with a concomitant decrease in type IIA and type I fibers in the Mstn(-/-) tibialis anterior and BF muscle. Functional electrical stimulation of Mstn(-/-) BF revealed increased fatigue susceptibility, supporting increased type IIB fiber content. Given the role of myocyte enhancer factor 2 (MEF2) in oxidative type I fiber formation, MEF2 levels in Mstn(-/-) tissue were quantified. Results revealed reduced MEF2C protein in Mstn(-/-) muscle and myoblast nuclear extracts. Reduced MEF2-DNA complex was also observed in electrophoretic mobility-shift assay using Mstn(-/-) nuclear extracts. Furthermore, reduced expression of MEF2 downstream target genes MLC1F and calcineurin were found in Mstn(-/-) muscle. Conversely, Mstn addition was sufficient to directly upregulate MLC promoter-enhancer activity in cultured myoblasts. Since high MyoD levels are seen in fast fibers, we analyzed MyoD levels in the muscle. In contrast to MEF2C, MyoD levels were increased in Mstn(-/ ) muscle. Together, these results suggest that while Mstn positively regulates MEF2C levels, it negatively regulates MyoD expression in muscle. We propose that Mstn could regulate fiber-type composition by regulating the expression of MEF2C and MyoD during myogenesis. PMID- 19129465 TI - TRPC6 mutations associated with focal segmental glomerulosclerosis cause constitutive activation of NFAT-dependent transcription. AB - Mutations in the canonical transient receptor potential channel TRPC6 lead to an autosomal dominant form of human kidney disease characterized histologically by focal and segmental glomerulosclerosis. Several of these mutations enhance the amplitude and duration of the channel current. However, the effect of these mutations on the downstream target of TRPC6, the nuclear factor of activated T cell (NFAT) transcription factors, has not been previously examined. Here we demonstrate that all three TRPC6 mutations previously shown to enhance channel activity lead to enhanced basal NFAT-mediated transcription in several cell lines, including cultured podocytes. These effects are dependent on channel activity and are dominant when mutants are coexpressed with wild-type TRPC6. While TRPC6 mutants do not demonstrate an increase in basal channel currents, a subset of cells expressing the R895C and E897K mutants have elevated basal calcium levels as measured by Fura-2 imaging. Activation of NFAT by TRPC6 mutants is blocked by inhibitors of calcineurin, calmodulin-dependent kinase II, and phosphatidylinositol 3-kinase. PP2 partially inhibits NFAT activation by mutant TRPC6 independently of Src, Yes, or Fyn. Differences in channel glycosylation and surface expression do not explain the ability of mutants to enhance NFAT activation. Taken together, these results identify the activation of the calcineurin-NFAT pathway as a potential mediator of focal segmental glomerulosclerosis. PMID- 19129466 TI - Comparison of intracellular cytokine flow cytometry and an enzyme immunoassay for evaluation of cellular immune response to active tuberculosis. AB - A prospective cross-sectional blinded study of 28 patients (21 male and 7 female patients; mean age, 44 years) with suspected active tuberculosis (TB) attending a TB and chest clinic is described. Blood was taken for immune cell enumeration, a whole-blood enzyme-linked immunosorbent assay (ELISA) for the detection of gamma interferon (IFN-gamma) by the QuantiFERON-TB Gold (QFT-G) assay, and intracellular cytokine flow cytometry (ICC) analysis; and sputum was simultaneously taken for bacteriological culture for Mycobacterium tuberculosis. Twelve healthy subjects were included as controls. The performance characteristics of the QFT-G and ICC assays for the detection of active TB were compared. Among the patients with active TB, we found (i) normal to slightly elevated peripheral CD4(+) and CD8(+) T-cell counts but a significant reduction in the number of NK cells; (ii) CD4(+) T cells were the major cell type producing IFN-gamma, a type 1 cytokine; (iii) small percentages of CD8(+) T cells were also primed for IFN-gamma production; (iv) the production of interleukin-4 (IL-4), a type 2 cytokine, was not prominent; and (v) the sensitivity and the specificity of the QFT-G assay were 88.2% and 18%, respectively, and those of the ICC assay were 94.1% and 36.4%, respectively. The specificities of the blood tests were likely underestimated due to cross-reaction to a non-M. tuberculosis mycobacterial infection and the lack of a confirmatory test that could be used to diagnose latent M. tuberculosis infection. Flow cytometry accurately locates the pool of immunological effector cells responsible for cytokine production during active TB. The ICC assay is an additional useful tool for the diagnosis of active TB. PMID- 19129467 TI - Relationship between human immunodeficiency virus type 1 coinfection, anemia, and levels and function of antibodies to variant surface antigens in pregnancy associated malaria. AB - Human immunodeficiency virus type 1 (HIV-1) coinfection decreases antibodies to variant surface antigens implicated in pregnancy-associated malaria (VSA-PAM) caused by Plasmodium falciparum. The effect of HIV-1 on antibody functions that may protect mothers from pregnancy-associated malaria is unknown. Sera from multigravid pregnant women with malaria and HIV-1 coinfection (n=58) or malaria alone (n=29) and from HIV-1-infected (n=102) or -uninfected (n=54) multigravidae without malaria were analyzed for anti-VSA-PAM antibodies by flow cytometry, the ability to inhibit adhesion to chondroitin sulfate A, or to opsonize CS2-infected erythrocytes for phagocytosis by THP-1 cells. In women with malaria, anti-VSA-PAM levels correlated better with opsonic activity (r=0.60) than with adhesion blocking activity (r=0.33). In univariate analysis, HIV-1 coinfection was associated with lower opsonic activity but not adhesion-blocking activity or anti VSA-PAM levels. Malaria-infected women with anemia (hemoglobin levels of <11.0 g/dl) had lower opsonic activity than nonanemic women (P=0.007) independent of HIV-1 status. By multivariate analysis, in malaria-infected women, anemia (but not HIV status) was associated with opsonic activity. In women without malaria, opsonic activity was not associated with either anemia or HIV-1 status. In multigravid pregnant women with malaria, impaired serum opsonic activity may contribute to anemia and possibly to the decreased immunity to pregnancy associated malaria associated with HIV-1. PMID- 19129468 TI - Humoral immune responses of white-tailed deer (Odocoileus virginianus) to Mycobacterium bovis BCG vaccination and experimental challenge with M. bovis. AB - Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple M. bovis antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (n=5), orally in liquid form (n=5), and subcutaneously (n=6) were evaluated over time after vaccination and after experimental challenge with virulent M. bovis and were compared to the responses by unvaccinated deer (n=6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple M. bovis antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials. PMID- 19129469 TI - Serological diagnosis of human herpes simplex virus type 1 and 2 infections by luciferase immunoprecipitation system assay. AB - Highly quantitative and high-throughput serological tests for evaluation of humoral responses to herpes simplex virus 1 (HSV-1) and HSV-2 are not available. The efficacy of luciferase immunoprecipitation system (LIPS) assays for antibody profiling and serologic diagnosis of HSV-1 and HSV-2 infection was investigated using a panel of five recombinant HSV antigens. Plasma samples from subjects seropositive for HSV-1 and/or HSV-2 or seronegative for HSV-1 and HSV-2 that had previously been analyzed by Western blotting and the Focus Plexus immunoassay were evaluated. The LIPS test measuring anti-gG1 antibody titers was 96% sensitive and 96% specific for detecting HSV-1 infection, compared with the Focus immunoassay, and was 92% sensitive and 96% specific, compared with Western blotting. The results for the anti-gG2 LIPS test for HSV-2 precisely matched those for Western blotting, with 100% sensitivity and 100% specificity, and showed robust antibody titers in all the HSV-2-infected samples that were over 1,000 times higher than those in HSV-2-negative or HSV-1-positive samples. Antibodies to three additional HSV-2 proteins, gB, gD, and ICP8, were detected in many of the HSV-1- and/or HSV-2-infected plasma samples and showed preferentially higher immunoreactivity in HSV-2-infected plasma. The titers of antibodies to these three HSV-2 antigens also significantly correlated with each other (R=0.75 to 0.81; P<0.0001). These studies indicate that the robust anti-gG1 and anti-gG2 antibody responses detected by LIPS assays are useful for HSV-1 and HSV-2 detection and suggest that profiling of antibody responses to a panel of HSV proteins may be useful for characterizing individual humoral responses to infection and for monitoring responses to vaccines. PMID- 19129470 TI - Simultaneous detection of Haemophilus influenzae type b polysaccharide-specific antibodies and Neisseria meningitidis serogroup A, C, Y, and W-135 polysaccharide specific antibodies in a fluorescent-bead-based multiplex immunoassay. AB - We expanded the meningococcal serogroup A, C, Y, and W-135 multiplex immunoassay (MIA) to simultaneously detect immunoglobulin type G antibodies directed toward Haemophilus influenzae type b polysaccharide (HibPS). The monoplex HibPS assay was compared to a HibPS-specific competitive enzyme-linked immunosorbent assay and showed a good correlation (R=0.96). Furthermore, no cross-reactivity between HibPS and the four meningococcal serogroups was detected. This pentaplex meningococcal Hib MIA is a useful tool to investigate serological responses toward different childhood PS vaccines. PMID- 19129471 TI - Toward diagnosing Leishmania infantum infection in asymptomatic dogs in an area where leishmaniasis is endemic. AB - The most frequently used diagnostic methods were compared in a longitudinal survey with Leishmania infantum-infected asymptomatic dogs from an area of Italy where leishmaniasis is endemic. In February and March 2005, 845 asymptomatic dogs were tested by an immunofluorescence antibody test (IFAT), a dipstick assay (DS), and an enzyme-linked immunosorbent assay (ELISA) for L. infantum and by IFAT for Ehrlichia canis. Dogs seronegative for L. infantum were further parasitologically evaluated by microscopic examination of lymph node tissues and PCR of skin samples. A total of 204 animals both serologically and parasitologically negative for L. infantum at the first sampling were enrolled in the trial and were further examined for canine leishmaniasis (CanL) and canine monocytic ehrlichiosis in November 2005 (i.e., the end of the first sandfly season) and March 2006 and 2007 (1- and 2-year follow-ups, respectively). At the initial screening, the overall rates of L. infantum seroprevalence were 9.5% by IFAT, 17.1% by ELISA, and 9.8% by DS and the overall rate of E. canis seroprevalence was 15%. The rates of concordance between the results of IFAT and DS were almost equal, whereas the rate of concordance between the results of IFAT and DS and those of the ELISA was lower. The results of the annual incidence of Leishmania infection were variable, depending on the test employed, with the highest values registered for PCR (i.e., 5.7% and 11.4% at the 1- and 2-year follow-ups, respectively), followed by ELISA, IFAT, and DS. Over the 2 years of observation, 55 animals (i.e., 26.9%) became positive for L. infantum by one or more diagnostic tests at different follow-up times, with 12.7% showing clinical signs related to CanL, while the remaining 87.3% were asymptomatic. A diagnostic scheme for assessment of the L. infantum infection status in asymptomatic dogs is suggested. PMID- 19129472 TI - Bacterial particle endocytosis by epithelial cells is selective and enhanced by tumor necrosis factor receptor ligands. AB - Bacterial pathogens use virulence strategies to invade epithelial barriers, but active processes of epithelial cells may also contribute to the endocytosis of microbial particles. To focus on the latter, we studied the uptake of fixed and fluorescently labeled bacterial particles in intestinal and bronchoepithelial cell cultures and found it to be enhanced in Caco-2BBe and NCI-H292 cells after treatment with tumor necrosis factor alpha and an agonist antibody against the lymphotoxin beta receptor. Confocal fluorescence microscopy, flow cytometry, and transmission electron microscopy revealed that Staphylococcus aureus and Yersinia enterocolitica were readily endocytosed, although there was scant uptake of Shigella sonnei, Salmonella enterica serovar Typhimurium, and Klebsiella pneumoniae particles. Endocytosed Staphylococcus was often associated with cytoplasmic claudin-4 vesicles; this was not found for Yersinia, suggesting that cytokine treatment upregulated two distinct endocytosis pathways. Interestingly, when Staphylococcus and Yersinia were coincubated with epithelial monolayers, the cells were unlikely to take up Yersinia unless they had also endocytosed large numbers of Staphylococcus particles, although the two bacteria were apparently processed in distinct compartments. Cytokine treatment induced an upregulation and redistribution of beta1 integrin to the apical surface of NCI-H292 cells; consistent with this effect, treatment with anti-beta1 integrin antibody blocked uptake of both Yersinia and Staphylococcus in NCI-H292 and Caco-2BBe cells. Our results suggest that capture of bacterial particles by mucosal epithelial cells is selective and that different endocytic mechanisms are enhanced by proinflammatory cytokines. PMID- 19129473 TI - Detection of anthrax toxin by an ultrasensitive immunoassay using europium nanoparticles. AB - We developed a europium nanoparticle-based immunoassay (ENIA) for the sensitive detection of anthrax protective antigen (PA). The ENIA exhibited a linear dose dependent pattern within the detection range of 0.01 to 100 ng/ml and was approximately 100-fold more sensitive than enzyme-linked immunosorbent assay (ELISA). False-positive results were not observed with serum samples from healthy adults, mouse plasma without PA, or plasma samples collected from mice injected with anthrax lethal factor or edema factor alone. For the detection of plasma samples spiked with PA, the detection sensitivities for ENIA and ELISA were 100% (11/11 samples) and 36.4% (4/11 samples), respectively. The assay exhibited a linear but qualitative correlation between the PA injected and the PA detected in murine blood (r=0.97731; P<0.0001). Anthrax PA was also detected in the circulation of mice infected with spores from a toxigenic Sterne-like strain of Bacillus anthracis, but only in the later stages of infection. These results indicate that the universal labeling technology based on europium nanoparticles and its application may provide a rapid and sensitive testing platform for clinical diagnosis and laboratory research. PMID- 19129474 TI - Cu, Zn superoxide dismutase and NADP(H) homeostasis are required for tolerance of endoplasmic reticulum stress in Saccharomyces cerevisiae. AB - Genome-wide screening for sensitivity to chronic endoplasmic reticulum (ER) stress induced by dithiothreitol and tunicamycin (TM) identified mutants deleted for Cu, Zn superoxide dismutase (SOD) function (SOD1, CCS1) or affected in NADPH generation via the pentose phosphate pathway (TKL1, RPE1). TM-induced ER stress led to an increase in cellular superoxide accumulation and an increase in SOD1 expression and Sod1p activity. Prior adaptation of the hac1 mutant deficient in the unfolded protein response (UPR) to the superoxide-generating agent paraquat reduced cell death under ER stress. Overexpression of the ER oxidoreductase Ero1p known to generate hydrogen peroxide in vitro, did not lead to increased superoxide levels in cells subjected to ER stress. The mutants lacking SOD1, TKL1, or RPE1 exhibited decreased UPR induction under ER stress. Sensitivity of the sod1 mutant to ER stress and decreased UPR induction was partially rescued by overexpression of TKL1 encoding transketolase. These data indicate an important role for SOD and cellular NADP(H) in cell survival during ER stress, and it is proposed that accumulation of superoxide affects NADP(H) homeostasis, leading to reduced UPR induction during ER stress. PMID- 19129475 TI - Heme oxygenase-1 contributes to an alternative macrophage activation profile induced by apoptotic cell supernatants. AB - Apoptotic cells (AC) are rapidly engulfed by professional phagocytes such as macrophages to avoid secondary necrosis and thus inflammation. Recognition of AC polarizes macrophages toward an anti-inflammatory phenotype, which shows homology to an alternatively activated M2 macrophage. However, mechanistic details provoking these phenotype alterations are incompletely understood. Here, we demonstrate a biphasic up-regulation of heme oxygenase-1 (HO-1), a protein that bears an antiapoptotic as well as an anti-inflammatory potential, in primary human macrophages, which were exposed to the supernatant of AC. Although the first phase of HO-1 induction at 6 h was accomplished by AC-derived sphingosine-1 phosphate (S1P) acting via S1P receptor 1, the second wave of HO-1 induction at 24 h was attributed to autocrine signaling of vascular endothelial growth factor A (VEGFA), whose expression and release were facilitated by S1P. Whereas VEGFA release from macrophages was signal transducer and activator of transcription (STAT) 1-dependent, vascular endothelial growth factor itself triggered STAT1/STAT3 heterodimer formation, which bound to and activated the HO-1 promoter. Knockdown of HO-1 proved its relevance in facilitating enhanced expression of the antiapoptotic proteins Bcl-2 and Bcl-X(L), as well as the anti inflammatory adenosine receptor A(2A). These findings suggest that HO-1, which is induced by AC-derived S1P, is critically involved in macrophage polarization toward an M2 phenotype. PMID- 19129476 TI - Oxysterol binding protein-related Protein 9 (ORP9) is a cholesterol transfer protein that regulates Golgi structure and function. AB - Oxysterol-binding protein (OSBP) and OSBP-related proteins (ORPs) constitute a large gene family that differentially localize to organellar membranes, reflecting a functional role in sterol signaling and/or transport. OSBP partitions between the endoplasmic reticulum (ER) and Golgi apparatus where it imparts sterol-dependent regulation of ceramide transport and sphingomyelin synthesis. ORP9L also is localized to the ER-Golgi, but its role in secretion and lipid transport is unknown. Here we demonstrate that ORP9L partitioning between the trans-Golgi/trans-Golgi network (TGN), and the ER is mediated by a phosphatidylinositol 4-phosphate (PI-4P)-specific PH domain and VAMP-associated protein (VAP), respectively. In vitro, both OSBP and ORP9L mediated PI-4P dependent cholesterol transport between liposomes, suggesting their primary in vivo function is sterol transfer between the Golgi and ER. Depletion of ORP9L by RNAi caused Golgi fragmentation, inhibition of vesicular somatitus virus glycoprotein transport from the ER and accumulation of cholesterol in endosomes/lysosomes. Complete cessation of protein transport and cell growth inhibition was achieved by inducible overexpression of ORP9S, a dominant negative variant lacking the PH domain. We conclude that ORP9 maintains the integrity of the early secretory pathway by mediating transport of sterols between the ER and trans-Golgi/TGN. PMID- 19129477 TI - Heterotrimerization of heat-shock factors 1 and 2 provides a transcriptional switch in response to distinct stimuli. AB - Organisms respond to circumstances threatening the cellular protein homeostasis by activation of heat-shock transcription factors (HSFs), which play important roles in stress resistance, development, and longevity. Of the four HSFs in vertebrates (HSF1-4), HSF1 is activated by stress, whereas HSF2 lacks intrinsic stress responsiveness. The mechanism by which HSF2 is recruited to stress inducible promoters and how HSF2 is activated is not known. However, changes in the HSF2 expression occur, coinciding with the functions of HSF2 in development. Here, we demonstrate that HSF1 and HSF2 form heterotrimers when bound to satellite III DNA in nuclear stress bodies, subnuclear structures in which HSF1 induces transcription. By depleting HSF2, we show that HSF1-HSF2 heterotrimerization is a mechanism regulating transcription. Upon stress, HSF2 DNA binding is HSF1 dependent. Intriguingly, when the elevated expression of HSF2 during development is mimicked, HSF2 binds to DNA and becomes transcriptionally competent. HSF2 activation leads to activation of also HSF1, revealing a functional interdependency that is mediated through the conserved trimerization domains of these factors. We propose that heterotrimerization of HSF1 and HSF2 integrates transcriptional activation in response to distinct stress and developmental stimuli. PMID- 19129478 TI - A fluorescently tagged C-terminal fragment of p47phox detects NADPH oxidase dynamics during phagocytosis. AB - The assembly of cytosolic p47(phox) and p67(phox) with flavocytochrome b(558) at the membrane is crucial for activating the leukocyte NADPH oxidase that generates superoxide for microbial killing. p47(phox) and p67(phox) are linked via a high affinity, tail-to-tail interaction involving a proline-rich region (PRR) and a C terminal SH3 domain (SH3b), respectively, in their C-termini. This interaction mediates p67(phox) translocation in neutrophils, but is not required for oxidase activity in model systems. Here we examined phagocytosis-induced NADPH oxidase assembly, showing the sequential recruitment of YFP-tagged p67(phox) to the phagosomal cup, and, after phagosome internalization, a probe for PI(3)P followed by a YFP-tagged fragment derived from the p47(phox) PRR. This fragment was recruited in a flavocytochrome b(558)-dependent, p67(phox)-specific, and PI(3)P independent manner. These findings indicate that p47PRR fragment probes the status of the p67(phox) SH3b domain and suggest that the p47(phox)/p67(phox) tail to-tail interaction is disrupted after oxidase assembly such that the p67(phox) SH3b domain becomes accessible. Superoxide generation was sustained within phagosomes, indicating that this change does not correlate with loss of enzyme activity. This study defines a sequence of events during phagocytosis-induced NADPH oxidase assembly and provides experimental evidence that intermolecular interactions within this complex are dynamic and modulated after assembly on phagosomes. PMID- 19129479 TI - Biochemical analyses of human IST1 and its function in cytokinesis. AB - The newly described yeast endosomal sorting complexes required for transport (ESCRT) protein increased sodium tolerance-1 (Ist1p) binds the late-acting ESCRT proteins Did2p/charged MVB protein (CHMP) 1 and Vps4p and exhibits synthetic vacuolar protein sorting defects when combined with mutations in the Vta1p/LIP5 Vps60p/CHMP5 complex. Here, we report that human IST1 also functions in the ESCRT pathway and is required for efficient abscission during HeLa cell cytokinesis. IST1 binding interactions with VPS4, CHMP1, LIP5, and ESCRT-I were characterized, and the IST1-VPS4 interaction was investigated in detail. Mutational and NMR spectroscopic studies revealed that the IST1 terminus contains two distinct MIT interacting motifs (MIM1 and MIM2) that wrap around and bind in different groves of the MIT helical bundle. IST1, CHMP1, and VPS4 were recruited to the midbodies of dividing cells, and depleting either IST1 or CHMP1 proteins blocked VPS4 recruitment and abscission. In contrast, IST1 depletion did not inhibit human immunodeficiency virus-1 budding. Thus, IST1 and CHMP1 act together to recruit and modulate specific VPS4 activities required during the final stages of cell division. PMID- 19129480 TI - Essential role of hIST1 in cytokinesis. AB - The last steps of multivesicular body (MVB) formation, human immunodeficiency virus (HIV)-1 budding and cytokinesis require a functional endosomal sorting complex required for transport (ESCRT) machinery to facilitate topologically equivalent membrane fission events. Increased sodium tolerance (IST) 1, a new positive modulator of the ESCRT pathway, has been described recently, but an essential function of this highly conserved protein has not been identified. Here, we describe the previously uncharacterized KIAA0174 as the human homologue of IST1 (hIST1), and we report its conserved interaction with VPS4, CHMP1A/B, and LIP5. We also identify a microtubule interacting and transport (MIT) domain interacting motif (MIM) in hIST1 that is necessary for its interaction with VPS4, LIP5 and other MIT domain-containing proteins, namely, MITD1, AMSH, UBPY, and Spastin. Importantly, hIST1 is essential for cytokinesis in mammalian cells but not for HIV-1 budding, thus providing a novel mechanism of functional diversification of the ESCRT machinery. Last, we show that the hIST1 MIM activity is essential for cytokinesis, suggesting possible mechanisms to explain the role of hIST1 in the last step of mammalian cell division. PMID- 19129481 TI - Centrosome/spindle pole-associated protein regulates cytokinesis via promoting the recruitment of MyoGEF to the central spindle. AB - Cooperative communications between the central spindle and the contractile ring are critical for the spatial and temporal regulation of cytokinesis. Here we report that MyoGEF, a guanine nucleotide exchange factor that localizes to the central spindle and cleavage furrow, interacts with centrosome/spindle pole associated protein (CSPP), which is concentrated at the spindle pole and central spindle during mitosis and cytokinesis. Both in vitro and in vivo pulldown assays show that MyoGEF interacts with CSPP. The C-terminus of MyoGEF and N-terminus of CSPP are required for their interaction. Immunofluorescence analysis indicates that MyoGEF and CSPP colocalize at the central spindle. Depletion of CSPP or MyoGEF by RNA-interference (RNAi) not only causes defects in mitosis and cytokinesis, such as metaphase arrest and furrow regression, but also mislocalization of nonmuscle myosin II with a phosphorylated myosin regulatory light chain (p-MRLC). Importantly, CSPP depletion by RNAi interferes with MyoGEF localization at the central spindle. Finally, MyoGEF interacts with ECT2, and RNAi-mediated depletion of MyoGEF leads to mislocalization of ECT2 and RhoA during cytokinesis. Therefore, we propose that CSPP interacts with and recruits MyoGEF to the central spindle, where MyoGEF contributes to the spatiotemporal regulation of cytokinesis. PMID- 19129482 TI - Role of atopic status in Toll-like receptor (TLR)7- and TLR9-mediated activation of human eosinophils. AB - Viral respiratory infections are increasingly implicated in allergic exacerbations. The mechanisms behind this are not known, but a virus-induced activation of eosinophils through TLRs could be involved. Herein, we investigated the expression and function of TLR7 and TLR9 in purified eosinophils from peripheral blood and assessed their role in allergic airway inflammation. Eosinophils expressed TLR7 and TLR9 proteins. Stimulation with the cognate ligands R-837 and CpG was found to prolong survival, up-regulate expression of CD11b and conversely down-regulate L-selectin expression, increase expression of the activation marker CD69, facilitate the chemotactic migration, and enhance IL 8 secretion by eosinophils. Also, CpG induced release of eosinophil-derived neurotoxin (EDN), and R-837 failed to do so. Analogously, eosinophils activated by CpG, but not R-837, promoted airway epithelial cell death and cytokine release. Priming with the allergic mediators histamine, IL-4, and most prominently IL-5, augmented the TLR-induced IL-8 and EDN secretion, revealing an ability to sensitize eosinophils for TLR7 and TLR9 activation. Moreover, the TLR responses of eosinophils were higher in allergic as compared with healthy subjects, manifested by an increase in IL-8 and EDN release. Correspondingly, allergic subjects displayed an elevated serum level of IL-5, suggesting increased IL-5-mediated priming. This study shows that activation via TLR7 and TLR9 affects several eosinophil functions and that the atopic status of the donor and the presence of a Th2-like cytokine milieu affect the outcome of the response. Thus, eosinophil activation via TLR7 and TLR9 might engender a link between viral infection and allergic exacerbations. PMID- 19129484 TI - Histone deacetylase inhibition improves differentiation of dendritic cells from leukemic blasts of patients with TEL/AML1-positive acute lymphoblastic leukemia. AB - Histone deacetylase inhibitors (HdI) could potentially improve the differentiation of leukemic dendritic cells (DC). Therefore, bone marrow samples from 100 children with acute lymphoblastic leukemia (ALL) were cultured in the cytokines TNF-alpha, GM-CSF, c-kit ligand, and fetal liver tyrosine kinase 3 ligand, with or without IL-3 and -4 and after administration of HdI valproic acid (VAL), suberoylanilide hydroxamic acid (SAHA), isobutyramid, or trichostatin A. Among the tested samples, 25 were positive for the chromosomal translocation t(12;21), encoding the fusion gene translocation ETS-like leukemia/acute myeloid leukemia 1 (TEL/AML1). SAHA increased CD83 expression of TEL/AML1-positive blasts in conditions without ILs, and SAHA and VAL increased the number of CD86(+)80(-) cells in the presence of ILs. VAL and isobutyramid supported the allostimulatory capacities of TEL/AML1-positive, leukemic DC; VAL and SAHA reduced those of TEL/AML1-negative DC. Cytotoxic T cells sensitized with leukemic DC produced more IFN-gamma and TNF-alpha upon presentation of the TEL/AML1 peptide. They also induced the cytotoxic lysis of nondifferentiated blasts, which was enhanced when TEL/AML1-positive DC had developed after addition of VAL or SAHA. Therefore, the use of HdI in the differentiation of leukemic DC from patients with TEL/AML1 positive ALL is recommended. PMID- 19129483 TI - Activation of nucleotide oligomerization domain 2 exacerbates a murine model of proteoglycan-induced arthritis. AB - In addition to its role in innate immunity, nucleotide oligomerization domain 2 (NOD2) has been shown to play a suppressive role in models of colitis. Notably, mutations in NOD2 cause the inherited granulomatous disease of the joints called Blau syndrome, thereby linking NOD2 with joint disease as well. However, the role of NOD2 in joint inflammation has not been clarified. We demonstrate here that NOD2 is functional within the mouse joint and promotes inflammation, as locally or systemically administered muramyl dipeptide (MDP; the NOD2 agonist) resulted in significant joint inflammation that was abolished in NOD2-deficient mice. We then sought to investigate the role of NOD2 in a mouse model of inflammatory arthritis dependent on adaptive immunity using TCR-transgenic mice whose T cells recognized the dominant epitope of proteoglycan (PG). Mice immunized with PG in the presence of MDP developed a more severe inflammatory arthritis and histopathology within the joints. Antigen-specific activation of splenocytes was enhanced by MDP with respect to IFN-gamma production, which would be consistent with the Th1-mediated disease in vivo. Intriguingly, NOD2 deficiency did not alter the PG-induced arthritis, indicating that NOD2 does not play an essential role in this model of joint disease when it is not activated by MDP. In conclusion, we demonstrate that in a model of inflammatory arthritis dependent on T and B cell priming, NOD2 activation potentiates disease. However, the absence of NOD2 does not alter the course of inflammatory arthritis, in contrast to models of intestinal inflammation. PMID- 19129485 TI - Superantigen-mediated differentiation of bovine monocytes into dendritic cells. AB - Although many effects of staphylococcal superantigens (SAg) on T cells are well established, less is known about their effects on APC. In this study, bovine PBMC were stimulated with a low dose of staphylococcal enterotoxin C1 (SEC1). The phenotype of adherent cells (Ac) derived from bovine PBMC cultured with SEC1 [SEC1-stimulated Ac (sAc)] for 192 h was CD14(-), CD68(-), CD163(-), dendritic cell (DC)-specific ICAM-3-grabbing nonintegrin(+), MHC class II (MHC II)(high), CD11a(low), CD11b(high), CD11c(high), and CD1b(high), suggesting these cells were dendritic cells (DC). SEC1 also induced transcription of the CXCL1, -2, and -3 family, CXCL6, CCL2, and CCL5 genes in sAc, which increased rapidly but returned to basal levels by 48 h. In contrast, increased transcription of CCL3, CCL8, and CXCL12, responsible for mononuclear cell migration and chronic inflammation, was sustained. In vitro cell migration assays showed vigorous migration of granulocytes, followed by migration of mononuclear cells. The autologous MLR showed that sAc induced a dose-dependent proliferation of CD4(+) T cells and an even stronger proliferation of CD8(+) T cells. This effect was inhibited or reduced by pretreatment with mAb to CD11b, MHC II, or MHC II plus CD18. These results indicate that stimulation of bovine PBMC by SAg induces differentiation of monocytes into DC. PMID- 19129486 TI - Differential impact of the CD45 juxtamembrane wedge on central and peripheral T cell receptor responses. AB - The cooperative activity of protein tyrosine kinases and phosphatases plays a central role in regulation of T cell receptor (TCR) signal strength. Perturbing this balance, and thus the threshold for TCR signals, has profound impacts on T cell development and function. We previously generated mice containing a point mutation in the juxtamembrane wedge of the receptor-like protein tyrosine phosphatase CD45. Demonstrating the critical negative regulatory function of the wedge, the CD45 E613R (WEDGE) mutation led to a lymphoproliferative disorder (LPD) and a lupus-like autoimmune syndrome. Using genetic, cellular, and biochemical approaches, we now demonstrate that the CD45 wedge influences T cell development and function. Consistent with increased TCR signal strength, WEDGE mice have augmented positive selection and enhanced sensitivity to the CD4 mediated disease experimental autoimmune encephalitis (EAE). These correspond with hyperresponsive calcium and pERK responses to TCR stimulation in thymocytes, but surprisingly, not in peripheral T cells, where these responses are actually depressed. Together, the data support a role for the CD45 wedge in regulation of T cell responses in vivo and suggest that its effects depend on cellular context. PMID- 19129487 TI - Photosynthetic metabolism of C3 plants shows highly cooperative regulation under changing environments: a systems biological analysis. AB - We studied the robustness of photosynthetic metabolism in the chloroplasts of C(3) plants under drought stress and at high CO(2) concentration conditions by using a method called Minimization of Metabolic Adjustment Dynamic Flux Balance Analysis (M_DFBA). Photosynthetic metabolism in the chloroplasts of C(3) plants applies highly cooperative regulation to minimize the fluctuation of metabolite concentration profiles in the face of transient perturbations. Our work suggests that highly cooperative regulation assures the robustness of the biological system and that there is closer cooperation under perturbation conditions than under normal conditions. This results in minimizing fluctuations in the profiles of metabolite concentrations, which is the key to maintaining a system's function. Our methods help in understanding such phenomena and the mechanisms of robustness for complex metabolic networks in dynamic processes. PMID- 19129488 TI - Nkx2-5 transactivates the Ets-related protein 71 gene and specifies an endothelial/endocardial fate in the developing embryo. AB - Recent studies support the existence of a common progenitor for the cardiac and endothelial cell lineages, but the underlying transcriptional networks responsible for specification of these cell fates remain unclear. Here we demonstrated that Ets-related protein 71 (Etsrp71), a newly discovered ETS family transcription factor, was a novel downstream target of the homeodomain protein, Nkx2-5. Using genetic mouse models and molecular biological techniques, we demonstrated that Nkx2-5 binds to an evolutionarily conserved Nkx2-5 response element in the Etsrp71 promoter and induces the Etsrp71 gene expression in vitro and in vivo. Etsrp71 was transiently expressed in the endocardium/endothelium of the developing embryo (E7.75-E9.5) and was extinguished during the latter stages of development. Using a gene disruption strategy, we found that Etsrp71 mutant embryos lacked endocardial/endothelial lineages and were nonviable. Moreover, using transgenic technologies and transcriptional and chromatin immunoprecipitation (ChIP) assays, we further established that Tie2 is a direct downstream target of Etsrp71. Collectively, our results uncover a novel functional role for Nkx2-5 and define a transcriptional network that specifies an endocardial/endothelial fate in the developing heart and embryo. PMID- 19129490 TI - Spectral methods in machine learning and new strategies for very large datasets. AB - Spectral methods are of fundamental importance in statistics and machine learning, because they underlie algorithms from classical principal components analysis to more recent approaches that exploit manifold structure. In most cases, the core technical problem can be reduced to computing a low-rank approximation to a positive-definite kernel. For the growing number of applications dealing with very large or high-dimensional datasets, however, the optimal approximation afforded by an exact spectral decomposition is too costly, because its complexity scales as the cube of either the number of training examples or their dimensionality. Motivated by such applications, we present here 2 new algorithms for the approximation of positive-semidefinite kernels, together with error bounds that improve on results in the literature. We approach this problem by seeking to determine, in an efficient manner, the most informative subset of our data relative to the kernel approximation task at hand. This leads to two new strategies based on the Nystrom method that are directly applicable to massive datasets. The first of these-based on sampling-leads to a randomized algorithm whereupon the kernel induces a probability distribution on its set of partitions, whereas the latter approach-based on sorting-provides for the selection of a partition in a deterministic way. We detail their numerical implementation and provide simulation results for a variety of representative problems in statistical data analysis, each of which demonstrates the improved performance of our approach relative to existing methods. PMID- 19129489 TI - Deadly competition between sibling bacterial colonies. AB - Bacteria can secrete a wide array of antibacterial compounds when competing with other bacteria for the same resources. Some of these compounds, such as bacteriocins, can affect bacteria of similar or closely related strains. In some cases, these secretions have been found to kill sibling cells that belong to the same colony. Here, we present experimental observations of competition between 2 sibling colonies of Paenibacillus dendritiformis grown on a low-nutrient agar gel. We find that neighboring colonies (growing from droplet inoculation) mutually inhibit growth through secretions that become lethal if the level exceeds a well-defined threshold. In contrast, within a single colony developing from a droplet inoculation, no growth inhibition is observed. However, growth inhibition and cell death are observed if material extracted from the agar between 2 growing colonies is introduced outside a growing single colony. To interpret the observations, we devised a simple mathematical model for the secretion of an antibacterial compound. Simulations of this model illustrate how secretions from neighboring colonies can be deadly, whereas secretions from a single colony growing from a droplet are not. PMID- 19129491 TI - Identification of Rhodococcus fascians cytokinins and their modus operandi to reshape the plant. AB - Decades ago, the importance of cytokinins (CKs) during Rhodococcus fascians pathology had been acknowledged, and an isopentenyltransferase gene had been characterized in the fas operon of the linear virulence plasmid, but hitherto, no specific CK(s) could be associated with virulence. We show that the CK receptors AHK3 and AHK4 of Arabidopsis thaliana are essential for symptom development, and that the CK perception machinery is induced upon infection, underlining its central role in the symptomatology. Three classical CKs [isopentenyladenine, trans-zeatin, and cis-zeatin (cZ)] and their 2-methylthio (2MeS)-derivatives were identified by CK profiling of both the pathogenic R. fascians strain D188 and its nonpathogenic derivative D188-5. However, the much higher CK levels in strain D188 suggest that the linear plasmid is responsible for the virulence-associated production. All R. fascians CKs were recognized by AHK3 and AHK4, and, although they individually provoked typical CK responses in several bioassays, the mixture of bacterial CKs exhibited clear synergistic effects. The cis- and 2MeS derivatives were poor substrates of the apoplastic CK oxidase/dehydrogenase enzymes and the latter were not cytotoxic at high concentrations. Consequently, the accumulating 2MeScZ (and cZ) in infected Arabidopsis tissue contribute to the continuous stimulation of tissue proliferation. Based on these results, we postulate that the R. fascians pathology is based on the local and persistent secretion of an array of CKs. PMID- 19129492 TI - Specific transbilayer translocation of dolichol-linked oligosaccharides by an endoplasmic reticulum flippase. AB - The oligosaccharide donor for protein N-glycosylation, Glc(3)Man(9)GlcNAc(2)-PP dolichol, is synthesized via a multistep pathway that starts on the cytoplasmic face of the endoplasmic reticulum (ER) and ends in the lumen where the glycosylation reaction occurs. This necessitates transbilayer translocation or flipping of the lipid intermediate Man(5)GlcNAc(2)-PP-dolichol (M5-DLO) across the ER membrane. The mechanism by which M5-DLO-or any other lipid-is flipped across the ER is unknown, except that specific transport proteins or flippases are required. We recently demonstrated M5-DLO flipping activity in proteoliposomes reconstituted from detergent-solubilized ER membrane proteins and showed that it was ATP-independent and required a trypsin-sensitive protein that sedimented at approximately 4S. By using an activity-enriched fraction devoid of glycerophospholipid flippase activity, we now report that M5-DLO is rapidly flipped in the reconstituted system with a time constant tau <2 min, whereas its triantennary structural isomer is flipped slowly with tau >200 min. DLOs larger than M5-DLO are also poorly translocated, with tau ranging from approximately 10 min to >200 min. We conclude that (i) the number and arrangement of mannoses in the DLO glycan has a profound effect on the ability of the DLO to be translocated by the flippase, (ii) glycan size per se does not dictate whether a DLO will be flipped, and (iii) the flippase is highly specific for M5-DLO. Our results suggest a simple structural model for the interaction between the DLO head group and the flippase. PMID- 19129493 TI - A G-protein gamma subunit mimic is a general antagonist of prion propagation in Saccharomyces cerevisiae. AB - The Gpg1 protein is a Ggamma subunit mimic implicated in the G-protein glucose signaling pathway in Saccharomyces cerevisiae, and its function is largely unknown. Here we report that Gpg1 blocks the maintenance of [PSI(+)], an aggregated prion form of the translation termination factor Sup35. Although the GPG1 gene is normally not expressed, over-expression of GPG1 inhibits propagation of not only [PSI(+)] but also [PIN(+)], [URE3] prions, and the toxic polyglutamine aggregate in S. cerevisiae. Over-expression of Gpg1 does not affect expression and activity of Hsp104, a protein-remodeling factor required for prion propagation, showing that Gpg1 does not target Hsp104 directly. Nevertheless, prion elimination by Gpg1 is weakened by over-expression of Hsp104. Importantly, Gpg1 protein is prone to self-aggregate and transiently colocalized with Sup35NM prion aggregates when expressed in [PSI(+)] cells. Genetic selection and characterization of loss-of-activity gpg1 mutations revealed that multiple mutations on the hydrophobic one-side surface of predicted alpha-helices of the Gpg1 protein hampered the activity. Prion elimination by Gpg1 is unaffected in the gpa2Delta and gpb1Delta strains lacking the supposed physiological G-protein partners of Gpg1. These findings suggest a general inhibitory interaction of the Gpg1 protein with other transmissible and nontransmissible amyloids, resulting in prion elimination. Assuming the ability of Gpg1 to form G-protein heterotrimeric complexes, Gpg1 is likely to play a versatile function of reversing the prion state and modulating the G-protein signaling pathway. PMID- 19129494 TI - Wnt/beta-catenin signaling is required for CNS, but not non-CNS, angiogenesis. AB - Despite the importance of CNS blood vessels, the molecular mechanisms that regulate CNS angiogenesis and blood-brain barrier (BBB) formation are largely unknown. Here we analyze the role of Wnt/beta-catenin signaling in regulating the formation of CNS blood vessels. First, through the analysis of TOP-Gal Wnt reporter mice, we identify that canonical Wnt/beta-catenin signaling is specifically activated in CNS, but not non-CNS, blood vessels during development. This activation correlates with the expression of different Wnt ligands by neural progenitor cells in distinct locations throughout the CNS, including Wnt7a and Wnt7b in ventral regions and Wnt1, Wnt3, Wnt3a, and Wnt4 in dorsal regions. Blockade of Wnt/beta-catenin signaling in vivo specifically disrupts CNS, but not non-CNS, angiogenesis. These defects include reduction in vessel number, loss of capillary beds, and the formation of hemorrhagic vascular malformations that remain adherent to the meninges. Furthermore, we demonstrate that Wnt/beta catenin signaling regulates the expression of the BBB-specific glucose transporter glut-1. Taken together these experiments reveal an essential role for Wnt/beta-catenin signaling in driving CNS-specific angiogenesis and provide molecular evidence that angiogenesis and BBB formation are in part linked. PMID- 19129495 TI - Nonequilibrium scale selection mechanism for columnar jointing. AB - Crack patterns in laboratory experiments on thick samples of drying cornstarch are geometrically similar to columnar joints in cooling lava found at geological sites such as the Giant's Causeway. We present measurements of the crack spacing from both laboratory and geological investigations of columnar jointing, and show how these data can be collapsed onto a single master scaling curve. This is due to the underlying mathematical similarity between theories for the cracking of solids induced by differential drying or by cooling. We use this theory to give a simple quantitative explanation of how these geometrically similar crack patterns arise from a single dynamical law rooted in the nonequilibrium nature of the phenomena. We also give scaling relations for the characteristic crack spacing in other limits consistent with our experiments and observations, and discuss the implications of our results for the control of crack patterns in thin and thick solid films. PMID- 19129496 TI - Rb/Cdk2/Cdk4 triple mutant mice elicit an alternative mechanism for regulation of the G1/S transition. AB - The G(1)/S-phase transition is a well-toned switch in the mammalian cell cycle. Cdk2, Cdk4, and the rate-limiting tumor suppressor retinoblastoma protein (Rb) have been studied in separate animal models, but interactions between the kinases and Rb in vivo have yet to be investigated. To further dissect the regulation of the G(1) to S-phase progression, we generated Cdk2(-/-)Cdk4(-/-)Rb(-/-) (TKO) mutant mice. TKO mice died at midgestation with major defects in the circulatory systems and displayed combined phenotypes of Rb(-/-) and Cdk2(-/-)Cdk4(-/-) mutants. However, TKO mouse embryonic fibroblasts were not only resistant to senescence and became immortal but displayed enhanced S-phase entry and proliferation rates similar to wild type. These effects were more remarkable in hypoxic compared with normoxic conditions. Interestingly, depletion of the pocket proteins by HPV-E7 or p107/p130 shRNA in the absence of Cdk2/Cdk4 elicited a mechanism for the G(1)/S regulation with increased levels of p27(Kip1) binding to Cdk1/cyclin E complexes. Our work indicates that the G(1)/S transition can be controlled in different ways depending on the situation, resembling a regulatory network. PMID- 19129497 TI - Importance of matrix habitats in maintaining biological diversity. PMID- 19129498 TI - Biodegradable dendritic positron-emitting nanoprobes for the noninvasive imaging of angiogenesis. AB - A biodegradable positron-emitting dendritic nanoprobe targeted at alpha(v)beta(3) integrin, a biological marker known to modulate angiogenesis, was developed for the noninvasive imaging of angiogenesis. The nanoprobe has a modular multivalent core-shell architecture consisting of a biodegradable heterobifunctional dendritic core chemoselectively functionalized with heterobifunctional polyethylene oxide (PEO) chains that form a protective shell, which imparts biological stealth and dictates the pharmacokinetics. Each of the 8 branches of the dendritic core was functionalized for labeling with radiohalogens. Placement of radioactive moieties at the core was designed to prevent in vivo dehalogenation, a potential problem for radiohalogens in imaging and therapy. Targeting peptides of cyclic arginine-glycine-aspartic acid (RGD) motifs were installed at the terminal ends of the PEO chains to enhance their accessibility to alpha(v)beta(3) integrin receptors. This nanoscale design enabled a 50-fold enhancement of the binding affinity to alpha(v)beta(3) integrin receptors with respect to the monovalent RGD peptide alone, from 10.40 nM to 0.18 nM IC(50). Cell-based assays of the (125)I-labeled dendritic nanoprobes using alpha(v)beta(3)-positive cells showed a 6-fold increase in alpha(v)beta(3) receptor-mediated endocytosis of the targeted nanoprobe compared with the nontargeted nanoprobe, whereas alpha(v)beta(3)-negative cells showed no enhancement of cell uptake over time. In vivo biodistribution studies of (76)Br labeled dendritic nanoprobes showed excellent bioavailability for the targeted and nontargeted nanoprobes. In vivo studies in a murine hindlimb ischemia model for angiogenesis revealed high specific accumulation of (76)Br-labeled dendritic nanoprobes targeted at alpha(v)beta(3) integrins in angiogenic muscles, allowing highly selective imaging of this critically important process. PMID- 19129499 TI - Chemical magnetoreception in birds: the radical pair mechanism. AB - Migratory birds travel vast distances each year, finding their way by various means, including a remarkable ability to perceive the Earth's magnetic field. Although it has been known for 40 years that birds possess a magnetic compass, avian magnetoreception is poorly understood at all levels from the primary biophysical detection events, signal transduction pathways and neurophysiology, to the processing of information in the brain. It has been proposed that the primary detector is a specialized ocular photoreceptor that plays host to magnetically sensitive photochemical reactions having radical pairs as fleeting intermediates. Here, we present a physical chemist's perspective on the "radical pair mechanism" of compass magnetoreception in birds. We outline the essential chemical requirements for detecting the direction of an Earth-strength approximately 50 microT magnetic field and comment on the likelihood that these might be satisfied in a biologically plausible receptor. Our survey concludes with a discussion of cryptochrome, the photoactive protein that has been put forward as the magnetoreceptor molecule. PMID- 19129500 TI - Trimorphic stepping stones pave the way to fungal virulence. PMID- 19129501 TI - Toward a genetics of cancer resistance. AB - Two of three humans never get cancer. Even the majority of heavy smokers remain cancer free. Is this a matter of chance, or are there cancer-resistant genotypes? Based on the evidence discussed, it would appear that evolution has favored a limited number of relatively common resistance genes that may nip incipient cancerous foci in the bud, i.e., to stop them at their inception. It is further suggested that resistance genes may act at the level of tissue organization in a dominant fashion. PMID- 19129503 TI - Evidence against rapid reversal of antiplatelet medications in acute intracerebral hemorrhage. PMID- 19129502 TI - Artificial ligand binding within the HIF2alpha PAS-B domain of the HIF2 transcription factor. AB - The hypoxia-inducible factor (HIF) basic helix-loop-helix Per-aryl hydrocarbon receptor nuclear translocator (ARNT)-Sim (bHLH-PAS) transcription factors are master regulators of the conserved molecular mechanism by which metazoans sense and respond to reductions in local oxygen concentrations. In humans, HIF is critically important for the sustained growth and metastasis of solid tumors. Here, we describe crystal structures of the heterodimer formed by the C-terminal PAS domains from the HIF2alpha and ARNT subunits of the HIF2 transcription factor, both in the absence and presence of an artificial ligand. Unexpectedly, the HIF2alpha PAS-B domain contains a large internal cavity that accommodates ligands identified from a small-molecule screen. Binding one of these ligands to HIF2alpha PAS-B modulates the affinity of the HIF2alpha:ARNT PAS-B heterodimer in vitro. Given the essential role of PAS domains in forming active HIF heterodimers, these results suggest a presently uncharacterized ligand-mediated mechanism for regulating HIF2 activity in endogenous and clinical settings. PMID- 19129504 TI - A new sensitive imaging biomarker for Parkinson disease? PMID- 19129505 TI - PML-IRIS in patients with HIV infection: clinical manifestations and treatment with steroids. AB - BACKGROUND: Progressive multifocal leukoencephalopathy (PML) is an opportunistic infection that develops in immunosuppressed patients with HIV infection. Paradoxically, some of these patients may develop PML during combined antiretroviral therapy in the setting of immune reconstitution. We describe the types of PML in relation to immune reconstitution inflammatory syndrome (IRIS) and the effects of steroid use in these patients. METHODS: We performed a retrospective review of the literature (1998 to 2007) and of all HIV-infected patients diagnosed with PML-IRIS at Johns Hopkins Hospital (2004 to 2007). We recorded information on clinical features, microbiologic and virological analysis, neuroimaging, pathology, treatment, and outcome. RESULTS: Of 54 patients with PML-IRIS, 36 developed PML and IRIS simultaneously (PML-s-IRIS) and 18 had worsening of preexisting PML (PML-d-IRIS) after the initiation of combined antiretroviral therapy. PML-IRIS developed between 1 week and 26 months after initiation of antiretroviral therapy. PML-d-IRIS patients developed IRIS earlier, had higher lesion loads on MRI of the brain, had shorter durations of survival, and had higher mortality rate compared to PML-s-IRIS patients. Twelve patients received treatment with steroids, of which five died and seven showed good neurologic recovery. Patients who survived had received steroids early after IRIS diagnosis for longer durations and had contrast enhancement on IRIS neuroimaging. CONCLUSIONS: Immune reconstitution following initiation of combined antiretroviral therapy may lead to activation of an inflammatory response to detectable or latent JC virus infection. Early and prolonged treatment with steroids may be useful in these patients but requires further investigation. PMID- 19129506 TI - Prior antiplatelet use does not affect hemorrhage growth or outcome after ICH. AB - OBJECTIVE: To examine whether antiplatelet medication use at onset of intracerebral hemorrhage (ICH) is associated with hemorrhage growth and outcome after spontaneous ICH using a large, prospectively collected database from a recent clinical trial. METHODS: The Cerebral Hemorrhage and NXY-059 Treatment trial was a randomized, placebo-controlled trial of NXY-059 after spontaneous ICH. We analyzed patients in the placebo arm, and correlated antiplatelet medication use at the time of ICH with initial ICH volumes, ICH growth in the first 72 hours, and modified Rankin Score at 90 days. Patients on oral anticoagulation were excluded. RESULTS: There were 282 patients included in this analysis, including 70 (24.8%) who were taking antiplatelet medications at ICH onset. Use of antiplatelet medications at ICH onset had no association with the volume of ICH at presentation, growth of ICH at 72 hours, initial edema volume, or edema growth. In multivariable analysis, there was no association of use of antiplatelet medications with any hemorrhage expansion (relative risk [RR] 0.85 [upper limit of confidence interval (UCI) 1.03], p = 0.16), hemorrhage expansion greater than 33% (RR 0.77 [UCI 1.18], p = 0.32), or clinical outcome at 90 days (odds ratio 0.67, 95% confidence interval 0.39-1.14, p = 0.14). CONCLUSIONS: Use of antiplatelet medications at intracerebral hemorrhage (ICH) onset is not associated with increased hemorrhage volumes, hemorrhage expansion, or clinical outcome at 90 days. These findings suggest that attempts to reverse antiplatelet medications after ICH may not be warranted. PMID- 19129507 TI - High-resolution diffusion tensor imaging in the substantia nigra of de novo Parkinson disease. AB - BACKGROUND: In the midbrain of patients with Parkinson disease (PD), there is a selective loss of dopaminergic neurons in the ventrolateral and caudal substantia nigra (SN). In a mouse model of PD, investigators have administered 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) and found that measures derived using diffusion tensor imaging (DTI) were correlated with the number of dopamine neurons lost following intoxication. METHODS: Twenty-eight subjects (14 with early stage, untreated PD and 14 age- and gender-matched controls) were studied with a high-resolution DTI protocol at 3 Tesla using an eight-channel phase array coil and parallel imaging to study specific segments of degeneration in the SN. Regions of interest were drawn in the rostral, middle, and caudal SN by two blinded and independent raters. RESULTS: Fractional anisotropy (FA) was reduced in the SN of subjects with PD compared with controls (p < 0.001). Post hoc analysis identified that reduced FA for patients with PD was greater in the caudal compared with the rostral region of interest (p < 0.00001). A receiver operator characteristic analysis in the caudal SN revealed that sensitivity and specificity were 100% for distinguishing patients with PD from healthy subjects. Findings were consistent across both raters. CONCLUSIONS: These findings provide evidence that high resolution diffusion tensor imaging in the substantia nigra distinguishes early stage, de novo patients with Parkinson disease (PD) from healthy individuals on a patient by patient basis and has the potential to serve as a noninvasive early biomarker for PD. PMID- 19129508 TI - Reduction of mouse egg surface integrin alpha9 subunit (ITGA9) reduces the egg's ability to support sperm-egg binding and fusion. AB - The involvement of egg integrins in mammalian sperm-egg interactions has been controversial, with data from integrin inhibitor studies contrasting with evidence from knockouts showing that specific integrin subunits are not essential for fertility. An alpha(4)/alpha(9) (ITGA4/ITGA9) integrin subfamily member has been implicated in fertilization but not extensively examined, so we tested the following three hypotheses: 1) an ITGA4/ITGA9 integrin participates in sperm-egg interactions, 2) short-term acute knockdown by RNA interference of integrin subunits would result in a fertilization phenotype differing from that of chronic depletion via knockout, and 3) detection of a fertilization phenotype is sensitive to in vitro fertilization (IVF) assay conditions. We show that mouse and human eggs express the alpha(9) integrin subunit (ITGA9). RNA interference mediated knockdown resulted in reduced levels of Itga9 mRNA and surface protein in mouse eggs. RNA interference attempts to knockdown ITGA9's likely beta partner, beta(1) (ITGB1), resulted in reduced Itgb1 mRNA but no reduction in ITGB1 surface protein. Therefore, studies using a function-blocking anti-ITGB1 antibody tested the hypothesis that ITGB1 participates in gamete interactions. Analyses of sperm-egg interactions with Itga9-knockdown eggs and anti-ITGB1 antibody-treated eggs in IVF assays using specific sperm:egg ratios revealed the following: 1) a reduction, but not complete loss, of sperm-egg binding and fusion was observed and 2) the reduction of sperm-egg binding and fusion was not detected in inseminations with high sperm:egg ratios. These data demonstrate that ITGA9 and ITGB1 participate in sperm-egg interactions but clearly are not the only molecules involved. This also shows that careful design of IVF parameters allows detection of deficiencies in gamete interactions. PMID- 19129510 TI - Comprehensive analysis of reproductive ADAMs: relationship of ADAM4 and ADAM6 with an ADAM complex required for fertilization in mice. AB - A Disintegrin And Metalloprotease (ADAM) family members expressed in male reproductive tissues are divided phylogenetically into three major groups. In the present study, we analyzed six ADAMs in one of the groups (ADAMs 4, 6, 24, 26, 29, and 30) of which function is largely unknown. Our results showed that most of the ADAMs undergo unique processing during sperm maturation and are located at the surface of sperm head. We found that the levels of ADAM4 and ADAM6 are dramatically reduced in Adam2 and Adam3 knockout sperm defective in various fertilization processes. We observed premature processing of ADAM4 in the Adam3 null mice. Furthermore, we obtained a result showing complex formation of ADAM6 with ADAM2 and ADAM3 in testis. Taken together, these results disclose involvement of ADAM4 and ADAM6 in a reproductive ADAM system that functions in fertilization. PMID- 19129509 TI - CDC14B acts through FZR1 (CDH1) to prevent meiotic maturation of mouse oocytes. AB - Meiotic maturation in oocytes is a prolonged process that is unique because of cell cycle arrests at prophase of meiosis I (MI) and at metaphase of meiosis II (MII). Fluctuations in cyclin-dependent kinase 1 (CDK1/CDC2A) activity govern meiotic progression, yet little is known about how these fluctuations are achieved. CDC14 is a highly conserved dual-specificity phosphatase that counteracts the function of proteins phosphorylated by CDK. Mammals contain two CDC14 homologs, CDC14A and CDC14B. We report that CDC14B localizes with the meiotic spindle in mouse oocytes, and (unlike somatic cells) it does not localize in the nucleolus. Oocytes that overexpress CDC14B are significantly delayed in resuming meiosis and fail to progress to MII, whereas oocytes depleted of CDC14B spontaneously resume meiosis under conditions that normally inhibit meiotic resumption. Depletion of FZR1 (CDH1), a regulatory subunit of the anaphase promoting complex/cyclosome that targets cyclin B1 (CCNB1) for ubiquitin-mediated proteolysis, partially restores normal timing of meiotic resumption in oocytes with excess CDC14B. These studies also reveal that experimentally altering CDC14B levels generates eggs with abnormal spindles and with chromosome alignment perturbations. Our data indicate that CDC14B is a negative regulator of meiotic resumption and may regulate MI in mouse oocytes. PMID- 19129511 TI - Gene birth, death, and divergence: the different scenarios of reproduction related gene evolution. AB - Reproductive genes are known to evolve more rapidly than genes expressed in other organs. In this paper we present an overview and bring some new data on the evolutionary study of reproduction-related genes by integrating phylogeny with gene genomic localization. We focus on the gene evolutionary processes of gene birth, death, and divergence. We show that phylogenetic gene birth is confirmed by gene location in genomes, which definitively localized the "place of birth" of new genes (such as Obox and KHDC1/DPPA5/ECAT1/OOEP gene families). By finding their "place of death" in genomes, it also demonstrates that ZP genes TGM4 and OVGP1 have been lost in certain species during vertebrate evolution. Moreover, in the case of gene divergence, comparison of gene locations across different genomes establishes orthologous relationships that are weakly supported by the phylogenetic tree. Specifically, genomic localization demonstrates that the fish and bird mtnr1c (Mel1C) receptor is orthologous to mammalian GPR50, and that ungulate genomes contain new seminal vesicle-specific BSP genes that are not present in other species. Overall, the phylogenomic approach to gene evolution presented in this paper offers more insight into gene function, such as species specific duplications for speciation, changes in gene expression due to gene divergence, and functional loss by gene death. PMID- 19129512 TI - Androgens upregulate cyp19a1b (aromatase B) gene expression in the brain of zebrafish (Danio rerio) through estrogen receptors. AB - The brain of teleosts is known for its strong aromatase expression, exhibiting unique features compared with other vertebrates. Among these features is the high sensitivity of aromatase B (the product of cyp19a1b) to estrogens. This effect involves the binding of estrogen receptors on an estrogen-responsive element (ERE) of the cyp19a1b promoter. Given the presence of potential androgen responsive elements (AREs) on this promoter, in vivo and in vitro effects of androgens were studied. Using immunohistochemistry and quantitative PCR on zebrafish embryos, we found that cyp19a1b is upregulated by testosterone, an aromatizable androgen, and by 5alpha-dihydrotestosterone (DHT), a nonaromatizable androgen, suggesting a potential androgenic regulation of cyp19a1b through androgen receptors (ARs). To assess a putative direct regulation of the cyp19a1b gene by ARs, we transfected U251MG cells with zebrafish AR together with a luciferase reporter gene driven by 3000 bp of the proximal cyp19a1b promoter containing the ERE and potential AREs. Interestingly, although zebrafish AR activated luciferase reporter genes controlled by AREs, they failed to induce the cyp19a1b-luciferase construct. These data suggest that the androgenic regulation of cyp19a1b does not involve AR. We further showed that regulation of the cyp19a1b gene by testosterone is, in fact, due to aromatization, whereas the effect of DHT involves conversion into 5alpha-androstane-3beta,17beta-diol (betadiol), a metabolite of DHT with known estrogenic activity. The blockage of the androgen regulation of cyp19a1b expression using antiestrogens further confirmed the involvement of estrogen receptors in mediating these effects. PMID- 19129513 TI - Dynamic nuclear organization of constitutive heterochromatin during fetal male germ cell development in mice. AB - In mice, male germ cells enter mitotic arrest beginning at 13.5 days postcoitum (dpc), and remain suspended in the G(0)/G(1) cell cycle stage until after birth. During this period, male germ cells undergo extensive epigenetic reprogramming, which is essential for their subsequent function as male gametes. A global reorganization and spatial clustering of constitutive heterochromatin has been implicated in epigenetic plasticity during cellular differentiation. Here, we have studied the dynamics of heterochromatin in fetal (12.5-19.5 dpc) and neonatal (4 days postpartum) male germ cells. We monitored constitutive heterochromatin-specific markers, and observed changes in the association of histone H3 trimethylation of lysine 9 (H3K9me3), binding of heterochromatin protein 1, and patterns of 4',6-diamino-2-phenylindole staining in pericentric regions of chromosomes, along with a coincident loss of chromocenters in fetal prospermatogonia during mitotic arrest. We also observed a transient loss of H3K9me3 associated with major and minor satellite repeat sequences, plus inactivation of histone methyltransferases (Suv39h1 and Suv39h2), and transient activation of histone demethylase (Jmjd2b) in these same cells. These epigenetic changes were correlated with relocation of centromeric regions toward the nuclear periphery in prospermatogonia during mitotic arrest. Taken together, these results show that constitutive heterochromatin undergoes dramatic reorganization during prespermatogenesis. We suggest that these dynamic changes in heterochromatin contribute to normal epigenetic reprogramming of the paternal genome in fetal prospermatogonia suspended in the G(0)/G(1) stage, and that this also represents an epigenomic state that is particularly amenable to reprogramming. PMID- 19129514 TI - Metabolic and mitochondrial dysfunction in early mouse embryos following maternal dietary protein intervention. AB - Dietary supply of nutrients, both periconception and during pregnancy, influence the growth and development of the fetus and offspring and their health into adult life. Despite the importance of research efforts surrounding the developmental origins of health and disease hypothesis, the biological mechanisms involved remain elusive. Mitochondria are of major importance in the oocyte and early embryo, particularly as a source of ATP generation, and perturbations in their function have been related to reduced embryo quality. The present study examined embryo development following periconception exposure of females to a high-protein diet (HPD) or a low-protein diet (LPD) relative to a medium-protein diet (MPD; control), and we hypothesized that perturbed mitochondrial metabolism in the mouse embryo may be responsible for the impaired embryo and fetal development reported by others. Although the rate of development to the blastocyst stage did not differ between diets, both the HPD and LPD reduced the number of inner cell mass cells in the blastocyst-stage embryo. Furthermore, mitochondrial membrane potential was reduced and mitochondrial calcium levels increased in the 2-cell embryo. Embryos from HPD females had elevated levels of reactive oxygen species and ADP concentrations, indicative of metabolic stress and, potentially, the uncoupling of oxidative phosphorylation, whereas embryos from LPD females had reduced mitochondrial clustering around the nucleus, suggestive of an overall quietening of metabolism. Thus, although periconception dietary supply of different levels of protein is permissive of development, mitochondrial metabolism is altered in the early embryo, and the nature of the perturbation differs between HPD and LPD exposure. PMID- 19129515 TI - The unfolded protein response contributes to preimplantation mouse embryo death in the DDK syndrome. AB - DDK syndrome is the polar-lethal embryonic death that occurs at the morula blastocyst transition when female mice of the DDK strain are mated with males from many other inbred strains (so-called alien males). Embryonic death is caused by incompatibility between a DDK oocyte factor and an alien male gene, both of which map to the Om locus on mouse chromosome 11. We compared global transcription patterns of DDK x DDK embryos (high viability) and DDK x C57BL/6 embryos (low viability) at the morula stage, approximately 24 h before any morphological manifestations of DDK syndrome are observed. Of the transcripts that are differentially more abundant in the DDK x C57BL/6 embryos, many are the products of genes induced by the "unfolded protein response." We confirmed by quantitative RT-PCR that a number of genes in this pathway are upregulated in the DDK x C57BL/6 embryos. Immunostaining of the endoplasmic reticulum (ER) marker BIP/GRP78 (immunoglobin-binding protein/glucose-regulated protein of 78 kDa), official symbol HSPA5, heat shock protein 5 revealed an accompanying abnormal HSPA5 accumulation and ER structure in the DDK x C57BL/6 embryos. Immunostaining for HERPUD1 (homocysteine-inducible, ER stress-inducible, ubiquitin-like domain member 1) and ATF4 (activating transcription factor 4) also revealed accumulation of these stress-response products. Our results indicate that the unfolded protein response is induced in embryos destined to die of DDK syndrome and that the embryonic death observed is associated with inability to resolve the associated ER stress. PMID- 19129516 TI - Survival role of locally produced acetylcholine in the bovine corpus luteum. AB - The present study was conducted to explore the source of acetylcholine (ACH) in the corpus luteum (CL) and to test our hypothesis of an antiapoptotic role of ACH in the bovine CL and, further, to investigate whether nerve growth factor (NGF), insulin-like growth factor 1 (IGF1), and transforming growth factor beta1 (TGFB1) influence the expression of choline acetyltransferase (CHAT), the biosynthetic enzyme of ACH, in cultured bovine luteal cells. Protein expression and immunolocalization of CHAT were carried out at different stages throughout the luteal phase and in cultured luteal and endothelial cells. ACH was measured in luteal tissue at the different luteal stages and in luteal cells cultured for 8 and 24 h. Cell viability and TUNEL assays were performed on cultured midluteal cells treated with or without tumor necrosis factor alpha (TNF)/interferon gamma (IFNG) in the presence of ACH and its muscarinic (atropine) and nicotinic (mecamylamine) receptor antagonists. The CL was devoid of cholinergic nerve fibers. CHAT immunostaining was evident in luteal, endothelial, and stromal cells in luteal tissue sections and in cultured luteal and endothelial cells. CHAT protein was expressed throughout the cycle without any significant changes. ACH concentration in luteal tissue was not changed during the luteal stages but increased over time and with increased cell numbers in luteal cell cultures. ACH increased cell viability and prevented cell death induced by TNF/IFNG. Atropine significantly attenuated ACH action, whereas mecamylamine had no effect. TNF/IFNG treatment downregulated CHAT expression, whereas NGF, IGF1, and TGFB1 upregulated CHAT expression, in cultured luteal cells. The overall findings strongly suggest a nonneural source and antiapoptotic role of ACH in the bovine CL. Locally produced ACH appears to be regulated by NGF, IGF1, and TGFB1. PMID- 19129517 TI - Correlates of hospitalization for alcohol-using methadone-maintained persons with physical health problems. AB - This cross-sectional study (n = 190) examined correlates of hospitalization for physical health problems among methadone maintenance therapy (MMT) clients with a history of alcohol abuse. The study was derived from baseline data collected for a longitudinal trial assessing the effect of motivational interviewing among alcohol-abusing adults undergoing MMT. The sample included clients who were 18-55 years of age, abusing alcohol, and receiving MMT from five large methadone maintenance clinics in the Los Angeles area. A structured questionnaire was used to collect the data. Correlates of hospitalization in logistic regression analysis included lack of social support, recent victimization, age of first alcohol use, chronic severe pain in the previous 6 months, not having children, and ethnicity. Identification of hospitalization risk factors among alcohol abusing MMT clients is a first step to developing risk-reducing interventions designed to lower hospitalization rates in this population. PMID- 19129518 TI - Recruitment of three generations of African American women into genetics research. AB - Successful outcomes for studies on health disparities depend on recruitment of research participants. Obtaining willing participants, protecting their rights, and acknowledging their contribution to research is as important as seeking answers to the study phenomena. Recruiting research participants can be an arduous process for investigators. Although literature has published participant recruitment methods, investigators sometimes underestimate the time and intensity required to attract eligible participants into research studies. This article reports on methods used to recruit 42 African American generational triads (grandmothers, mothers, and granddaughters) into a hypertension genetics study, the lessons learned, and suggestions for successful recruitment. PMID- 19129519 TI - Cultural Competency in Baccalaureate Nursing Education. AB - This article presents the work of the American Association of Colleges of Nursing Advisory Committee in developing an integrated curriculum for cultural competence in baccalaureate nursing education. Five competencies for Cultural Competency in Baccalaureate Nursing Education were developed with supporting rationale. Suggestions for developing and implementing curricula for cultural competency, teaching content, specific integrative learning strategies, methods for evaluating nursing students' cultural competence and recommendations for effective implementation of the integrated curriculum are presented. PMID- 19129521 TI - Images in clinical medicine. Medication-induced gingival hypertrophy. PMID- 19129522 TI - Money and the changing culture of medicine. PMID- 19129520 TI - Deletion of IKZF1 and prognosis in acute lymphoblastic leukemia. AB - BACKGROUND: Despite best current therapy, up to 20% of pediatric patients with acute lymphoblastic leukemia (ALL) have a relapse. Recent genomewide analyses have identified a high frequency of DNA copy-number abnormalities in ALL, but the prognostic implications of these abnormalities have not been defined. METHODS: We studied a cohort of 221 children with high-risk B-cell-progenitor ALL with the use of single-nucleotide-polymorphism microarrays, transcriptional profiling, and resequencing of samples obtained at diagnosis. Children with known very-high-risk ALL subtypes (i.e., BCR-ABL1-positive ALL, hypodiploid ALL, and ALL in infants) were excluded from this cohort. A copy-number abnormality was identified as a predictor of poor outcome, and it was then tested in an independent validation cohort of 258 patients with B-cell-progenitor ALL. RESULTS: More than 50 recurring copy-number abnormalities were identified, most commonly involving genes that encode regulators of B-cell development (in 66.8% of patients in the original cohort); PAX5 was involved in 31.7% and IKZF1 in 28.6% of patients. Using copy-number abnormalities, we identified a predictor of poor outcome that was validated in the independent validation cohort. This predictor was strongly associated with alteration of IKZF1, a gene that encodes the lymphoid transcription factor IKAROS. The gene-expression signature of the group of patients with a poor outcome revealed increased expression of hematopoietic stem cell genes and reduced expression of B-cell-lineage genes, and it was similar to the signature of BCR-ABL1-positive ALL, another high-risk subtype of ALL with a high frequency of IKZF1 deletion. CONCLUSIONS: Genetic alteration of IKZF1 is associated with a very poor outcome in B-cell-progenitor ALL. PMID- 19129523 TI - The Neurontin legacy--marketing through misinformation and manipulation. PMID- 19129524 TI - Toward the elimination of schistosomiasis. PMID- 19129526 TI - A strategy to control transmission of Schistosoma japonicum in China. AB - BACKGROUND: Schistosoma japonicum causes an infection involving humans, livestock, and snails and is a significant cause of morbidity in China. METHODS: We evaluated a comprehensive control strategy in two intervention villages and two control villages along Poyang Lake in the southeastern province of Jiangxi, where annual synchronous chemotherapy is routinely used. New interventions, implemented from 2005 through 2007, included removing cattle from snail-infested grasslands, providing farmers with mechanized farm equipment, improving sanitation by supplying tap water and building lavatories and latrines, providing boats with fecal-matter containers, and implementing an intensive health education program. During the intervention period, we observed changes in S. japonicum infection in humans, measured the rate of infection in snails, and tested the infectivity of lake water in mice. RESULTS: After three transmission seasons, the rate of infection in humans decreased to less than 1.0% in the intervention villages, from 11.3% to 0.7% in one village and from 4.0% to 0.9% in the other (P<0.001 for both comparisons). The rate of infection in humans in control villages fluctuated but remained at baseline levels. In intervention villages, the percentage of sampling sites with infected snails decreased from 2.2% to 0.1% in one grassland area and from 0.3% to no infection in the other (P<0.001 for both comparisons). The rate of infection in mice after exposure to lake water decreased from 79% to no infection (P<0.001). CONCLUSIONS: A comprehensive control strategy based on interventions to reduce the rate of transmission of S. japonicum infection from cattle and humans to snails was highly effective. These interventions have been adopted as the national strategy to control schistosomiasis in China. PMID- 19129525 TI - Timing of elective repeat cesarean delivery at term and neonatal outcomes. AB - BACKGROUND: Because of increased rates of respiratory complications, elective cesarean delivery is discouraged before 39 weeks of gestation unless there is evidence of fetal lung maturity. We assessed associations between elective cesarean delivery at term (37 weeks of gestation or longer) but before 39 weeks of gestation and neonatal outcomes. METHODS: We studied a cohort of consecutive patients undergoing repeat cesarean sections performed at 19 centers of the Eunice Kennedy Shriver National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network from 1999 through 2002. Women with viable singleton pregnancies delivered electively (i.e., before the onset of labor and without any recognized indications for delivery before 39 weeks of gestation) were included. The primary outcome was the composite of neonatal death and any of several adverse events, including respiratory complications, treated hypoglycemia, newborn sepsis, and admission to the neonatal intensive care unit (ICU). RESULTS: Of 24,077 repeat cesarean deliveries at term, 13,258 were performed electively; of these, 35.8% were performed before 39 completed weeks of gestation (6.3% at 37 weeks and 29.5% at 38 weeks) and 49.1% at 39 weeks of gestation. One neonatal death occurred. As compared with births at 39 weeks, births at 37 weeks and at 38 weeks were associated with an increased risk of the primary outcome (adjusted odds ratio for births at 37 weeks, 2.1; 95% confidence interval [CI], 1.7 to 2.5; adjusted odds ratio for births at 38 weeks, 1.5; 95% CI, 1.3 to 1.7; P for trend <0.001). The rates of adverse respiratory outcomes, mechanical ventilation, newborn sepsis, hypoglycemia, admission to the neonatal ICU, and hospitalization for 5 days or more were increased by a factor of 1.8 to 4.2 for births at 37 weeks and 1.3 to 2.1 for births at 38 weeks. CONCLUSIONS: Elective repeat cesarean delivery before 39 weeks of gestation is common and is associated with respiratory and other adverse neonatal outcomes. PMID- 19129528 TI - Mortality attributable to smoking in China. AB - BACKGROUND: Smoking is a risk factor for many diseases and has been increasingly prevalent in economically developing regions of the world. We aimed to estimate the number of deaths attributable to smoking in China. METHODS: We conducted a large, prospective cohort study in a nationally representative sample of 169,871 Chinese adults who were 40 years of age or older. Investigators for the China National Hypertension Survey collected data on smoking and other risk factors at a baseline examination in 1991 using a standard protocol. Follow-up evaluation was conducted in 1999 and 2000, with a response rate of 93.4%. We used multivariable-adjusted relative risk, prevalence of smoking, mortality, and population size in each age group, stratified according to sex, to calculate the number of deaths attributable to smoking in 2005. RESULTS: There was a significant, dose-response association between pack-years smoked and death from any cause in both men and women after adjustment for multiple risk factors (P<0.001 for trend). We estimated that in 2005, a total of 673,000 deaths (95% confidence interval [CI], 564,700 to 781,400) were attributable to smoking in China: 538,200 (95% CI, 455,800 to 620,600) among men and 134,800 (95% CI, 108,900 to 160,800) among women. The leading causes of smoking-related deaths were as follows: cancer, 268,200 (95% CI, 214,500 to 321,900); cardiovascular disease, 146,200 (95% CI, 79,200 to 213,100); and respiratory disease, 66,800 (95% CI, 20,300 to 113,300). CONCLUSIONS: Our study documents that smoking is a major risk factor for mortality in China. Continued strengthening of national programs and initiatives for smoking prevention and cessation is needed to reduce smoking-related deaths in China. PMID- 19129529 TI - Clinical practice. Vitiligo. PMID- 19129527 TI - Arterial blood gases and oxygen content in climbers on Mount Everest. AB - BACKGROUND: The level of environmental hypobaric hypoxia that affects climbers at the summit of Mount Everest (8848 m [29,029 ft]) is close to the limit of tolerance by humans. We performed direct field measurements of arterial blood gases in climbers breathing ambient air on Mount Everest. METHODS: We obtained samples of arterial blood from 10 climbers during their ascent to and descent from the summit of Mount Everest. The partial pressures of arterial oxygen (PaO(2)) and carbon dioxide (PaCO(2)), pH, and hemoglobin and lactate concentrations were measured. The arterial oxygen saturation (SaO(2)), bicarbonate concentration, base excess, and alveolar-arterial oxygen difference were calculated. RESULTS: PaO(2) fell with increasing altitude, whereas SaO(2) was relatively stable. The hemoglobin concentration increased such that the oxygen content of arterial blood was maintained at or above sea-level values until the climbers reached an elevation of 7100 m (23,294 ft). In four samples taken at 8400 m (27,559 ft)--at which altitude the barometric pressure was 272 mm Hg (36.3 kPa)--the mean PaO(2) in subjects breathing ambient air was 24.6 mm Hg (3.28 kPa), with a range of 19.1 to 29.5 mm Hg (2.55 to 3.93 kPa). The mean PaCO(2) was 13.3 mm Hg (1.77 kPa), with a range of 10.3 to 15.7 mm Hg (1.37 to 2.09 kPa). At 8400 m, the mean arterial oxygen content was 26% lower than it was at 7100 m (145.8 ml per liter as compared with 197.1 ml per liter). The mean calculated alveolar-arterial oxygen difference was 5.4 mm Hg (0.72 kPa). CONCLUSIONS: The elevated alveolar-arterial oxygen difference that is seen in subjects who are in conditions of extreme hypoxia may represent a degree of subclinical high-altitude pulmonary edema or a functional limitation in pulmonary diffusion. PMID- 19129530 TI - Images in clinical medicine. Diagnosis of uterine rupture on CT. PMID- 19129531 TI - Case records of the Massachusetts General Hospital. Case 1-2009. A 57-year-old man with progressive cognitive decline. PMID- 19129532 TI - Making small risks even smaller. PMID- 19129533 TI - Tiotropium in chronic obstructive pulmonary disease. PMID- 19129534 TI - Defibrillators in heart failure and quality of life. PMID- 19129535 TI - Magnesium sulfate for the prevention of cerebral palsy. PMID- 19129536 TI - Continuous glucose monitoring and type 1 diabetes. PMID- 19129537 TI - Monoclonal antibody therapy and non-Hodgkin's lymphoma. PMID- 19129538 TI - Elimination of antibodies to recombinant enzyme in Pompe's disease. PMID- 19129539 TI - Glycemic control in patients with insulinoma treated with everolimus. PMID- 19129540 TI - Inhibition of monocyte-derived inflammatory cytokines by IL-25 occurs via p38 Map kinase-dependent induction of Socs-3. AB - IL-25, a member of the IL-17 cytokine family, is known to enhance Th2-like responses associated with increased serum levels of IgE, IgG1, IgA, blood eosinophilia, and eosinophilic infiltrates in various tissues. However, IL-25 also abrogates inflammatory responses driven by Th17 cells. However, the cell types that respond to IL-25 and the mechanisms by which IL-25 differentially regulates immune reactions are not well explored. To identify potential targets of IL-25, we initially examined IL-25 receptor (IL-25R) in human peripheral blood cells. IL-25R was predominantly expressed by CD14(+) cells. We next assessed the functional role of IL-25 in modulating the response of CD14(+) cells to various inflammatory signals. CD14(+) cells responded to IL-25 by down-regulating the synthesis of inflammatory cytokines induced by toll-like receptor (TLR) ligands and inflammatory cytokines. Inhibition of cytokine response by IL-25 occurred via a p38 Map kinase-driven Socs-3-dependent mechanism. In vivo, IL-25 inhibited monocyte-derived cytokines and protected against LPS-induced lethal endotoxemia in mice. These data indicate that IL-25 is a negative regulator of monocyte proinflammatory cytokine responses, which may have therapeutic implications. PMID- 19129541 TI - Mesenchymal stem/progenitor cells promote the reconstitution of exogenous hematopoietic stem cells in Fancg-/- mice in vivo. AB - Fanconi anemia (FA) is a heterogeneous genetic disorder characterized by bone marrow failure and complex congenital anomalies. Although mutations in FA genes result in a characteristic phenotype in the hematopoietic stem/progenitor cells (HSPCs), little is known about the consequences of a nonfunctional FA pathway in other stem/progenitor cell compartments. Given the intense functional interactions between HSPCs and the mesenchymal microenvironment, we investigated the FA pathway on the cellular functions of murine mesenchymal stem/progenitor cells (MSPCs) and their interactions with HSPCs in vitro and in vivo. Here, we show that loss of the murine homologue of FANCG (Fancg) results in a defect in MSPC proliferation and in their ability to support the adhesion and engraftment of murine syngeneic HSPCs in vitro or in vivo. Transplantation of wild-type (WT) but not Fancg(-/-) MSPCs into the tibiae of Fancg(-/-) recipient mice enhances the HSPC engraftment kinetics, the BM cellularity, and the number of progenitors per tibia of WT HSPCs injected into lethally irradiated Fancg(-/-) recipients. Collectively, these data show that FA proteins are required in the BM microenvironment to maintain normal hematopoiesis and provide genetic and quantitative evidence that adoptive transfer of WT MSPCs enhances hematopoietic stem cell engraftment. PMID- 19129542 TI - The genomic architecture of segmental duplications and associated copy number variants in dogs. AB - Structural variation is an important and abundant source of genetic and phenotypic variation. Here we describe the first systematic and genome-wide analysis of segmental duplications and associated copy number variants (CNVs) in the modern domesticated dog, Canis familiaris, which exhibits considerable morphological, physiological, and behavioral variation. Through computational analyses of the publicly available canine reference sequence, we estimate that segmental duplications comprise approximately 4.21% of the canine genome. Segmental duplications overlap 841 genes and are significantly enriched for specific biological functions such as immunity and defense and KRAB box transcription factors. We designed high-density tiling arrays spanning all predicted segmental duplications and performed aCGH in a panel of 17 breeds and a gray wolf. In total, we identified 3583 CNVs, approximately 68% of which were found in two or more samples that map to 678 unique regions. CNVs span 429 genes that are involved in a wide variety of biological processes such as olfaction, immunity, and gene regulation. Our results provide insight into mechanisms of canine genome evolution and generate a valuable resource for future evolutionary and phenotypic studies. PMID- 19129544 TI - Biblical framings of and responses to spousal violence in the narratives of abused christian women. AB - Through narrative, the authors examine the coping activities of three Christian women to learn how they used religion to organize actions intended to end spouse abuse. Findings suggest an apparent creative attempt to stop violation. Respondents formed coping strategies from biblical archetypes that resembled their oppression and clued a method to its end. However, their responses encouraged ownership of spousal change and impeded removal of violation. Given these confines, the authors address why religion became an important resource. A discussion on legal and cultural views surrounding abused women's responses and the contextual factors that limit but do not preclude acts of subversion is given. PMID- 19129545 TI - The relationships between social goals, skills, and strategies and their effect on aggressive behavior among adolescents. AB - It is widely accepted that conflict-related goals, skills, and strategies are linked. Yet it is rarely explored how these factors relate to each other and how they jointly promote or inhibit aggressive behaviors. The aim of this study is to provide answers to these questions. Data were derived from a structured questionnaire administered to 660 male and female adolescents of an average age of 14.99 years from two urban schools in northern Israel. Findings show that goals, skills, and strategies that promote or inhibit violence are positively interrelated. Furthermore, negative association was found between violence promoting and inhibiting goals, skills, and strategies. Gender differences were also analyzed. It has been found that boys display aggressive behavior more frequently then girls. Findings also show that the rate of violence promoting goals, skills, and strategies is higher among boys than among girls, whereas that of violence inhibiting ones are higher among girls than among boys. Yet when controlling the effects of goals, skills, and strategies, girls demonstrate aggressive behavior more frequently than boys. These research findings are discussed and conceptualized within the theoretical framework of social adjustment. PMID- 19129546 TI - An itemized approach to assessing behavioral and psychological symptoms in dementia. AB - In an effort to obtain more detailed clinical information regarding behavioral and psychology symptoms in dementia, we submitted an existing, comprehensive measure of behavioral and psychology symptoms in dementia (Neuropsychiatric Inventory) to an alternate itemized scoring system. One hundred twenty-four caregivers of patients with dementia (mean Mini-Mental State Examination=22.6) rated the frequency of individual symptoms across all domains of the measure. Internal reliability and factor structures for all domains were analyzed to assess the stability of this scoring approach. Internal consistency alphas for each domain ranged from .57 to .91. Alpha reliability for the total inventory was .96. Results indicate an itemized approach to assessing behavioral and psychological symptoms in dementia among patients with mild-to-moderate dementia can be reliable, has the power to capture multiple features of neuropsychiatric symptoms, and can produce a rich neurobehavioral profile adding valuable information to the diagnosis and treatment of these patients. PMID- 19129543 TI - Cell-type selective chromatin remodeling defines the active subset of FOXA1-bound enhancers. AB - Selective activity of a specific set of enhancers defines tissue-specific gene transcription. The pioneer factor FOXA1 has been shown to induce functional enhancer competency through chromatin openings. We have previously found that FOXA1 is recruited to thousands of regions across the genome of a given cell type. Here, we monitored the chromatin structure at FOXA1 binding sites on a chromosome-wide scale using formaldehyde assisted isolation of regulatory elements (FAIRE). Surprisingly, we find that a significant fraction of FOXA1 bound sites have a relatively closed chromatin conformation linked to a shift of the epigenetic signature toward repressive histone marks. Importantly, these sites are not correlated with gene expression in a given cell type suggesting that FOXA1 is required, but not sufficient, for the functional activity of bound enhancers. Interestingly, we find that a significant proportion of the inactive FOXA1-bound regulatory sites in one cell type are actually functional in another cellular context. We found that at least half of the FOXA1 binding sites from a given cell type are shared with another cell lineage. Mechanisms that restrict the activity of shared FOXA1-bound enhancers likely play a significant role in defining the cell-type-specific functions of FOXA1. PMID- 19129548 TI - [Third round: modern psychiatry: has it ever existed in Hungary?]. PMID- 19129547 TI - Quetiapine effective in treatment of inappropriate sexual behavior of lewy body disease with predominant frontal lobe signs. AB - Dementia of Lewy body disease is the second most common degenerative cause of dementia after Alzheimer's disease, among all the dementias. The core features are a progressive dementia, fluctuations in cognitive functions, visual hallucinations, and spontaneous parkinsonism. Rapid eye movement sleep behavior disorder, severe neuroleptic sensitivity, and low dopamine transporter uptake in basal ganglia are other suggestive features. Behavioral abnormalities are commonly present in the form of aggressive behavior, irritability, and uninhibited behaviors. These are mostly seen in the advanced stages of dementia. However, inappropriate sexual behavior is uncommonly seen in such cases. Three types of inappropriate sexual behaviors commonly found in cases of dementia are sex talks, sexual acts, and implied sexual acts. Such inappropriate sexual behaviors have not been described adequately in dementia of Lewy body disease. We report inappropriate sexual behaviors in a case of dementia of Lewy body disease, which improved rapidly after treatment with quetiapine. PMID- 19129549 TI - [Neuropsychological impairments in adult attention deficit hyperactivity disorder: a literature review]. AB - INTRODUCTION: Attention Deficit Hyperactivity Disorder (ADHD) is one of the most frequently diagnosed childhood psychiatric disorders. Based on various empirical estimates, symptoms of ADHD persist into adulthood in about 30-50% of the childhood cases. METHOD: The objective of the present investigation was to conduct a systematic review of neuropsychological studies to characterize executive and attentional performance in subjects with adult ADHD and to examine whether these neuropsychological tools are useful to diagnose the disorder. 29 relevant publications were identified from 1993 to 2007 via a comprehensive research in Medline and Pubmed databases. RESULTS: The neuropsychological characteristics of adults with ADHD are comparable to those described in the pediatric population. Results indicate that in ADHD the frontal lobe neurocognitive functions, such as attention, planning, behaviour inhibition and problem solving, are the ones primarily impaired in the disorder. DISCUSSION: A better understanding of the nature and extent of the impairment of the cognitive functions involved in the disorder would offer opportunities for the development of disease-specific cognitive therapies and psychopharmacological therapies for patients with ADHD. Furthermore, a finer specification of the neurocognitive profile in adult ADHD might be of great importance as a sensitive marker for the efficacy of pharmaceutical agents. PMID- 19129550 TI - [Gambling and pathological gambling in Hungary]. AB - Gambling and pathological gambling are typically out of focus research area in Hungary. Besides the little evidence available in this field, it can be stated that both society and policy decision makers ignore the problems regarding this phenomenon. Gambling is rather considered to be a segment of the market than a behavior with possible harmful consequences or a mental and behavior disorder . The author reviews current knowledge on gambling and pathological gambling, including market data, research results and treatment options and highlights the most important steps which would be necessary in order to develop this research area. PMID- 19129551 TI - [P300 in mild cognitive impairment and in dementia]. AB - OBJECTIVES: The late component of the event-related potential (ERP), P300 is a marker for cognitive brain functions. The objective of this study was to examine P300 parameters in dementia and in mild cognitive impairment (MCI), to examine whether there is a correlation between the changes in P300 parameters and the type and severity of dementia, and to determine P300 abnormalities in MCI in relation to the presence/absence of CT/MRI abnormalities. METHOD: Auditory event related potentials, P300 were recorded in 28 MCI patients (14 of them with normal CT/MRI findings and 10 subjects with mild ventricular enlargement). In another group of patients, 31 demented patients were examined, of whom 17 patients were diagnosed with Alzheimer's dementia (AD), and 14 patients with vascular dementia (VD). Forty healthy volunteers served as the control group. RESULTS: Mean P300 latency was significantly increased for both demented patient-groups. We found that prolongation of P300 latency was correlated with the severity of dementia. Mean P300 amplitude was significantly decreased in both groups of demented patients. In MCI, the mean latency of P300 was significantly longer among patients with mild cerebral atrophy compared to control volunteers and no significant changes were found for MCI patients with normal CT/MRI findings. Mean P300 amplitude was decreased only in demented patients. CONCLUSIONS: The prolongation of P300 latency was significant among patients with both vascular and Alzheimer's dementia, and also among MCI patients with mild cerebral atrophy. The severity of dementia is positively correlated with P300 latency; however, this prolongation is independent of the type of dementia. The structural brain changes in MCI are related to P300 latency prolongation and thus may indicate an increased risk for developing dementia in MCI patients. PMID- 19129552 TI - [Screening childhood behavior problems using short questionnaires I.: the Hungarian version of the Strengths and Difficulties Questionnaire]. AB - INTRODUCTION: The Strengths and Difficulties Questionnaire (SDQ) is a short questionnaire suitable for screening childhood behavior problems. Completing the questionnaire requires 5 minutes of parents' or teachers' time. The scales of the original version showed good agreement with relevant scales of the much longer Child Behavior Checklist (CBCL). In this study, we report the use of the Hungarian version of the SDQ and results of a comparison between the scales of the SDQ and the CBCL. METHODS: Questionnaire data were collected from the parents of 156 six-year-old children from a community sample. Of the 156 children, 89 were participants of the longitudinal Budapest Infant-Parent Study (BIPS). RESULTS: Internal consistency of the Hungarian SDQ scales were moderate satisfactory (0.43-0.70), correlations between the relevant SDQ and CBCL scales were as expected (0.41-0.65), showing that the short questionnaire was equally suitable for the detection of problems. In our six-year-old age group, the mean of the total problem scores (11.0) was much higher than the means measured in other Western European countries and North-America. However, in developing countries, such as Brazil or China, levels were higher and closer to the Hungarian mean score. CONCLUSIONS: These initial Hungarian data confirm international experience with the SDQ, i.e., psychometric indices and the distribution of scale scores across the sexes are consistent with the results of foreign studies. On the other hand, a cautious generalization of our results indicates a higher level of problems in Hungary than in Western Europe and other developed countries. We think that following further collection of normative data the Hungarian version of the SDQ, which will take only a few minutes to complete, will be suitable for assessing the mental health of children and adolescents, and for the quick screening of problematic cases. PMID- 19129553 TI - [The birth of biological therapy in Hungary: the story of Laszlo Meduna's first patient receiving convulsive therapy]. AB - Convulsive treatment was introduced to psychiatry by Laszlo Meduna, a Hungarian neuropsychiatrist. In his autobiography, Meduna gave a detailed description of his first patient who underwent convulsive therapy. According to Meduna's recollections, this patient was L. Zoltan, who after 4 years of fluctuating catatonic stupor received several sessions of camphor-induced convulsive therapy resulting in full remission and discharge from the institution. In this communication, the authors reconstruct L Zoltan's case history from the original case notes, which were recovered from the Archives of the National Institute of Psychiatry and Neurology, Budapest. The case notes show that L. Zoltan received courses of convulsive treatment between 1934 and 1937, first induced with camphor and then with cardiazol. After the first course of treatment the catatonic stupor was resolved and the psychotic symptoms subsided. However, this incomplete remission lasted only for a few months and was followed by a relapse. Despite further courses of convulsive therapy, L. Zoltan never became symptom free again, could never be discharged and died in the institute in 1945. The authors attempt to explain possible reasons for the discrepancies found between Meduna's account and L. Zoltan's case notes. PMID- 19129554 TI - [Knowledge about and attitudes toward suicide in the Hungarian general population and in the helping professions]. AB - In the framework of an international suicide prevention program, we examined suicide-related attitudes and knowledge using the questionnaire constructed by Hubbard-McIntosh in the Hungarian population (N=4238) and among 231 helping professional. Our results show that proper information and false assumptions as well as attitudes toward suicide (refusal or acceptance of the right to commit suicide in general and exceptional situations in life) are closely related to several characteristics of the individual's socio-demographic position, to geographical location, to depression, and to the personal involvement in suicide related events. In line with our expectations, the level of knowledge of helping professionals is higher than that of the general population. Compared to the general population, a significantly higher proportion of helping professionals deny the right to commit suicide; however, in relation to a particular situation of life - e.g. terminal illness - their opinion is similar to that of the general population. PMID- 19129556 TI - Comparative clinical and transcriptomal profiles of breast cancer between French and South Mediterranean patients show minor but significative biological differences. AB - BACKGROUND: In Western countries, breast cancer incidence and mortality are higher than in Mediterranean countries. These differences have been ascribed to environmental factors but also to late-stage diagnostic and biological specific characteristics. PATIENTS AND METHODS: Between September 2002 and September 2005, we collected clinical data by phone counselling 180 French and Mediterranean breast cancer patients and performed microarray experiments. RESULTS: Characteristics of breast cancer in patients from Lebanon, Tunisia and Morocco were more aggressive (more SBR grade III and positive node invasion) and patients were 10 years younger at diagnosis. Sixteen differentially expressed genes such as MMP9, VEGF, PHB1, BRCA1, TFAP2C, GJA1 and TFF1 were also found. Additionally, an up-regulation of cytokeratins KRT8 and KRT18 may indicate a luminal B subtype in "South" (Lebanon, Tunisia and Morocco) tumors while "North" (France) tumors may more frequently be luminal A type. CONCLUSION: This study allowed the identification of specific clinical and transcriptomic parameters in patients from South Mediterranean countries. PMID- 19129555 TI - Secondary structure at a hot spot for DNA methylation in DNA from human breast cancers. AB - The VNTR at c-Ha-ras resides in a hotspot for DNA methylation on chromosome 11 in human tumors, where it is flanked by two MspI restriction sites. We have investigated the nature of the MspI site polymorphism at the c-Ha-ras VNTR observed in variety of tumors including breast cancer.We find that the MspI site 5' to the VNTR is present in a Non-B DNA structure with single-strand character that renders it accessible to bisulfite modification under native conditions, while the MspI site 3' to the VNTR appears to reside in a normal B-form structure that is inaccessible to bisulfite. The non-B DNA structure accounts for the observed polymorphism since MspI cannot cleave single-stranded DNA and control experiments show that the MspI sites were neither mutated nor abnormally methylated. Southern blotting showed that structural polymorphism was present in tumor DNA and tumor adjacent normal tissue DNA but absent from lymphocyte DNA from the same patients. We conclude that the non-B DNA structural polymorphism detected in human tumors near the c-Ha-ras VNTR is a self-perpetuating epigenetic mark that manifests itself spontaneously during breast carcinogenesis in a methylation hot spot. PMID- 19129557 TI - Tissue microarrays of human tumor xenografts: characterization of proteins involved in migration and angiogenesis for applications in the development of targeted anticancer agents. AB - As new target-directed anticancer agents emerge, preclinical efficacy studies need to integrate target-driven model systems. This approach to drug development requires rapid and reliable characterization of the new targets in established tumor models, such as xenografts and cell lines. Here, we have applied tissue microarray technology to patient-derived, re-growable human tumor xenografts. We have profiled the expression of five proteins involved in cell migration and/or angiogenesis: vascular endothelial growth factor (VEGF), matrix metalloproteinase 1 (MMP1), protease activated receptor (PAR1), cathepsin B, and beta1 integrin in a panel of over 150 tumors and compared their expression levels to available patient outcome data. For each protein, several target overexpressing xenografts were identified. They represent a subset of tumor models prone to respond to specific inhibitors and are available for future preclinical efficacy trials. In a "proof of concept" experiment, we have employed tissue microarrays to select in vivo models for therapy and for the analysis of molecular changes occurring after treatment with the anti-VEGF antibody HuMV833 and gemcitabine. Whereas the less angiogenic pancreatic cancer PAXF736 model proved to be resistant, the highly vascularized PAXF546 xenograft responded to therapy. Parallel analysis of arrayed biopsies from the different treatment groups revealed a down-regulation of Ki-67 and VEGF, an altered tissue morphology, and a decreased vessel density. Our results demonstrate the multiple advantages of xenograft tissue microarrays for preclinical drug development. PMID- 19129558 TI - Improved effector function of leukemia-specific T-lymphocyte clones trained with AML-derived dendritic cells. AB - Recently it was shown that myeloid leukemic cells can be induced to differentiate into leukemia-derived dendritic cells (DCleu), regaining the stimulatory capacity of professional DCs while presenting the leukemic antigen repertoire. But so far, the induced antileukemic T-cell responses have varied in specificity and efficacy, or have even mediated opposite effects. In an attempt to further characterize the DC/DCleu induced T-cell response pattern, immunoscope spectratyping, a novel and powerful tool to detect T-cell receptor (TCR) rearrangements was used in combination with functional flow cytometry and non radioactive fluorolysis assays. Human leucocyte antigen (HLA) matched donor T cells were repeatedly stimulated, either with leukemic blasts (French-American British, FAB M4eo) or the corresponding blast-derived DCs. Functional comparison revealed no significant difference in their T-cell stimulatory capacity, while the DC/DCleu fraction favored T-cells with a higher lytic activity, comprising a higher proportion of T-memory CD45R0+ cells. Stimulation with blasts and DC/DCleu induced a similar TCR restriction pattern, while stimulation with DC/DCleu favored the CD4 T-cell subset and seemed to cause a higher grade of restriction. In conclusion, a combined strategy using spectratyping with functional tests might not only provide useful information about the specificity and efficacy of the induced T-cell response, but also pave the way to gain effective T-cell clones for therapeutic use. PMID- 19129560 TI - Probiotics--the health friendly gut bacteria. PMID- 19129559 TI - Genomic changes of the 55 kDa subunit of DNA polymerase epsilon in human breast cancer. AB - BACKGROUND: DNA polymerases (Pols) represent potential candidates for cancer genes because of their central functions in DNA metabolism. Defects of some DNA Pols have shown cancer associations, but data on DNA polymerase (Pol) epsilon is limited. MATERIALS AND METHODS: Twenty-four human breast cancer DNA samples and four control DNA samples were examined for possible mutation in the entire coding region of the 55 kDa small subunit of the human DNA Pol epsilon gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of the DNA and sequence analysis. In addition, 20 control DNAs were studied with PCR-SSCP for the end of intron 18 and exon 19 region. RESULTS: An AATT deletion was found at one location in intron 18 in 2 out of the 24 breast cancer cases (8%), but in none of the control cases. In addition, a single base transition was found in the cancer DNAs in intron 14, but the same changes were also found in the control DNAs, suggesting polymorphism. CONCLUSION: Specific changes might occur in the 55 kDa small subunit DNA sequence of DNA Pol epsilon in breast cancer. The deletion at the region of intron-exon junction may not affect the protein code, but could potentially influence splicing efficiency and expression levels, possibly impairing the function of Pol epsilon DNA. PMID- 19129561 TI - Management issues among children living with HIV looking ahead. PMID- 19129562 TI - Smoking among youth in East Timor-Leste: what are the dynamics? PMID- 19129563 TI - Prevalence and correlates of current cigarette smoking among adolescents in East Timor-Leste. AB - OBJECTIVES: To determine the prevalence and correlates for current cigarette smoking. DESIGN: Secondary analysis of the East Timor-Leste Global Youth Tobacco Survey conducted in 2006. SETTING: Public and private schools registered with the Ministry of Education. PARTICIPANTS: A two-stage cluster sample of 1790 students in Grades 7 to 9. Schools were selected with probability proportional to enrolment size, and classes were randomly selected in each school. All students in selected classes were eligible to participate in the survey. The school and student response rates were 96.0% and 84.5%, respectively. MAIN OUTCOME MEASURE: Prevalence of current cigarette smoking. RESULTS: Out of 1790 adolescents, 52.1% were of ages less than 15 years, 51.8% were males, 42.8% reported having some pocket money in a month, and 72.7% had at least a parent who was a smoker. Prevalence of current cigarette smoking was 40.3%. Current smokers also reported having bought cigarettes from peddlers (32.4%), someone bought for them (16.7%), got from someone older (13.7%), borrowed (13.3%), and stole (3.4%). Males were more likely to be smokers than females (59.0% versus 19.3%). Factors positively associated with current smoking were: parental smoking; closest friend smoking; amount of pocket money; and exposure to anti-tobacco messages. CONCLUSIONS: East Timor has one of the highest prevalence of cigarette smoking among adolescents. The fact that exposure to anti-tobacco messages was associated with being a smoker may be evidence suggesting that anti-tobacco messages, especially from tobacco-related industry, may have unintended consequences. PMID- 19129564 TI - Viral pathogens of acute lower respiratory tract infection in China. AB - OBJECTIVES: To document the viral etiology of acute lower respiratory tract infection (ALRIs) in Chinese children. SETTING: Children Hospital, Zhejiang University, China. STUDY DESIGN: Cross-sectional. PARTICIPANTS: 34885 children with ALRI between January 2001 to December 2006. METHODS: Nasopharyngeal aspirates were collected from all subjects. Respiratory syncytial virus (RSV), adenovirus (ADV), type 1 to 3 parainfluenza viruses (PIV), and type A and B influenza virus (Flu) were detected by direct immunofluorescence. RESULTS: Viruses were identified in 32.3% cases, including RSV (23.6%), PIV 3 (4.3%), Flu A (2.0%), ADV (1.7%), PIV I (0.6%), Flu B (0.2%) and PIV 2 (0.1%). RSV and PIV 3 predominated in younger children while Flu A and Flu B predominated in older children (P<0.001, respectively). PIV 1 was more prevalent in children aged 1 to 3 years. The peak frequency of RSV, PIV 3 and Flu A were in early spring, June to August, and August and September, respectively. Flu B had a peak in the winter and spring. Adenovirus infections occurred in all seasons with a relatively constant frequency. CONCLUSIONS: Viruses are an important cause of ALRIs in Chinese children constituting 1/3 of total cases. RSV is the most common pathogen. PMID- 19129565 TI - Fetal alcohol spectrum disorder. AB - BACKGROUND: Maternal alcohol use during pregnancy leads to fetal alcohol spectrum disorder (FASD) in their children. FASD is characterized by typical facial features, growth retardation, intellectual dysfunction and behavioral problems. JUSTIFICATION: Alcohol is neurotoxic to the brain during the developmental stage. Behavioral problems in children with FASD start at an early age and progress to adulthood. It is an important preventable cause of intellectual dysfunction and behavioral problems. This article reviews current prevalence, clinical features, pathogenesis and differential diagnosis of FASD. It also highlights the need for physicians to be aware of this condition. SEARCH STRATEGY: Articles were searched on the internet using fetal alcohol syndrome, fetal alcohol spectrum disorders, women and alcohol. Following links were used to locate journals; EBSCO, OVID, Science Direct, PubMed and NIAAA. MAIN CONCLUSIONS: Alcohol consumption during pregnancy can lead to a spectrum of deficits. Though physical features are essential to make the diagnosis of FAS, it is important to note that neurocognitive and behavioural deficits can be present in the absence of physical features (alcohol related neurodevelopmental disorder or ARND). Because there is no known safe amount of alcohol consumption during pregnancy, abstinence from alcohol for women who are pregnant or planning a pregnancy must be strongly advised. PMID- 19129567 TI - Hypertonic saline nebulization for bronchiolitis. PMID- 19129568 TI - Age profile of neonatal deaths. AB - Neonatal survival has emerged as the key to further reduction in child mortality. Distribution of neonatal deaths in first week of life in the community is poorly understood. In a cross sectional survey covering a population of 13,16,681, information was collected retrospectively for one-year reference period on 30,473 births, 1,521 neonatal deaths and 2,218 infant deaths from five rural sites in India. Of all neonatal deaths, 39.3% occurred on first day of life, and 56.8 % during the first three days. The study highlights importance of first three days as the most hazardous phase in life and provides evidence base for postnatal care guidelines. PMID- 19129569 TI - Intraarticular triamcinolone in juvenile idiopathic arthritis. AB - Thirty seven children with juvenile idiopathic arthritis (JIA) who were treated with one or more intra-articular triamcinolone acetonide (TA) injections were evaluated. Ninety five joints were injected with a total number of 125 injections. Complete remission of the joint inflammation lasting at least 6 months was obtained in 62 of 95 injections (65%). Treatment of the joint contractures was successful in 35 of 51 joints (69%). In patients with oligoarthritis, 21 of 26 injected joints (81%) were in full remission at six months. The 6-month remission was significantly lower in the other subtypes of JIA (P<0.01), 41 of 69 (59%) injected joints, when compared to oligoarticular patients. Intra-articular TA injection is an effective and safe therapy for inflammatory joint disease in JIA, particularly in the oligoarticular form. PMID- 19129570 TI - Acute disseminated encephalomyelitis presenting as acute psychotic disorder. AB - A 5 year old boy and a 10 year old girl presented with acute onset of psychotic disorder, which occurred one week after an upper respiratory infection. MRI images of brain were consistent with the diagnosis of acute disseminated encephalomyelitis (ADEM) in both cases. ADEM is one of the differential diagnoses to be considered when acute psychotic disorder occurs during childhood. PMID- 19129571 TI - Acute hemorrhagic edema of infancy. AB - We describe an 18 month old girl with acute hemorrhagic edema of infancy, with palpable erythemaous purpuric rash on face, upper and lower limbs and ear lobules associated with edema and leukocytoclastic vasculitis. There were no systemic complications. PMID- 19129572 TI - Primary amebic meningoencephalitis. AB - We report an 8 month-old infant with primary amebic meningoencephalitis (PAME) due to Naegleria fowleri. The child was treated with amphotericin B, chloramphenicol and rifampicin for 3 weeks. PAME is an almost universally fatal condition with very few survivors till date. Our patient was one of the rare survivors who recovered after treatment and was discharged without any residual neurological deficit. PMID- 19129573 TI - Kleine-Levin syndrome. PMID- 19129574 TI - Disseminated cysticercosis in an infant. PMID- 19129575 TI - Misuse of corticosteroids in infants of rural Tamilnadu. PMID- 19129587 TI - New vanadium(IV) and titanium(IV) oxyfluorotellurates(IV): V2Te2O7F2 and TiTeO3F2. AB - As part of a continuing study of oxyfluorotellurates(IV), materials likely to present interesting nonlinear optical properties, two new phases, titanium(IV) tellurium(IV) trioxide difluoride, TiTeO(3)F(2), and divanadium(IV) ditellurium(IV) heptaoxide difluoride, V(2)Te(2)O(7)F(2), have been characterized and present, respectively, titanium and vanadium in the tetravalent state. The TiTeO(3)F(2) structure is based on linear double rows of TiO(3)F(3) polyhedra sharing vertices. These rows are connected to adjacent rows via two vertices of Te(2)O(5) bipolyhedra. The Te, Ti, one F and two O atoms are on general positions, with one O and F statistically occupying the same site with half occupancy for each anion. One O and one F occupy sites with .m. symmetry. The V(2)Te(2)O(7)F(2) structure consists of zigzag chains of VO(4)F(2) octahedra alternately sharing O-O and F-F edges. These chains are connected via Te(2)O(5) bipolyhedra, forming independent mixed layers. The Te, V, one F and three O atoms are on general positions while one O atom occupies a site of ?overline{1} symmetry. In both phases, the electronic lone pair E of the Te(IV) atom is stereochemically active. A full O/F anionic ordering is observed in V(2)Te(2)O(7)F(2), but in TiTeO(3)F(2) one of the six anionic sites is occupied by half oxygen and half fluorine, all the others being strictly ordered. These compounds represent new members of a growing family of oxyfluorotellurates(IV), including the recently characterized members of formula MTeO(3)F, M being a trivalent cation. As was true for the previous members, they are characterized by an unusually high thermal and chemical stability in relation to the absence of direct Te-F bonds. PMID- 19129588 TI - Redetermination of bis[mu(3)-1,3,5-triamino-1,3,5-trideoxy-cis-inositolato(3 )]tribismuth(III) trichloride hexahydrate. AB - The crystal structure of the title compound, [Bi(3)(C(6)H(12)N(3)O(3))(2)]Cl(3).6H(2)O, which was described in the space group R3 [Hegetschweiler, Ghisletta & Gramlich (1993). Inorg. Chem. 32, 2699-2704], has been redetermined in the revised space group R32 as suggested by Marsh [Acta Cryst. (2002), B58, 893-899]. Accordingly, the significant difference in the Bi-N bond distances of 2.43 (2) and 2.71 (1) A, as noted in the previous study, proved to be an artifact. As a consequence, the [Bi(3)(H(-3)taci)(2)]Cl(6/3) entity (taci is 1,3,5-triamino-1,3,5-trideoxy-cis-inositol) adopts D(3) symmetry and the three Bi atoms lie on C(2) axes with equal Bi-N bond distances of 2.636 (3) A. PMID- 19129589 TI - Manganese(II) and cobalt(II) complexes of 1,4-bis(diphenylphosphinoyl)butane. AB - The title complexes, catena-poly[[[diaquadiethanolmanganese(II)]-mu-1,4 bis(diphenylphosphinoyl)butane-kappa(2)O:O'] dinitrate 1,4 bis(diphenylphosphinoyl)butane solvate], {[Mn(C(2)H(6)O)(2)(C(28)H(28)O(2)P(2))(H(2)O)(2)](NO(3))(2).C(28)H(28)O(2)P(2)}(n , (I), and catena-poly[[[diaquadiethanolcobalt(II)]-mu-1,4 bis(diphenylphosphinoyl)butane-kappa(2)O:O'] dinitrate 1,4 bis(diphenylphosphinoyl)butane solvate], {[Co(C(2)H(6)O)(2)(C(28)H(28)O(2)P(2))(H(2)O)(2)](NO(3))(2).C(28)H(28)O(2)P(2)}(n , (II), are isostructural and centrosymmetric, with the M(II) ions at centres of inversion. The coordination geometry is octahedral, with each metal ion coordinated by two trans ethanol molecules, two trans water molecules and two bridging 1,4-bis(diphenylphosphinoyl)butane ligands which link the coordination centres to form one-dimensional polymeric chains. Parallel chains are linked by hydrogen bonds to uncoordinated 1,4-bis(diphenylphosphinoyl)butane molecules, which are bisected by a centre of inversion. Further hydrogen bonds, weak C-H...O interactions to nitrate anions, and weak C-H...pi interactions serve to stabilize the structure. This study reports a development of the coordination chemistry of bis(diphenylphosphinoyl)alkanes, with the first reported structures of complexes of the first-row transition metals with 1,4-bis(diphenylphosphinoyl)butane. PMID- 19129590 TI - Bis[S-6-(2,2:6',2''-terpyridin-4'-yloxy)hexyl thioacetate]manganese(II) bis(hexafluorophosphate). AB - The structure of a manganese(II) complex of terpyridine functionalized with acetylsulfanyl-terminated hexyloxy chains, [Mn(C(23)H(25)N(3)O(2)S)(2)](PF(6))(2), is described. This type of complex is of interest in the study of single-molecule transport properties in open-shell systems. The manganese coordination environment is distorted octahedral but, importantly, with no larger deviations from the idealized geometry than those observed for other metal-terpyridine complexes. The Mn-N bond lengths range from 2.192 (2) to 2.272 (3) A. The title compound crystallizes with the cation and anions all on general positions, with the hexafluorophosphate anions exhibiting orientational disorder. When compared with other bis-terpyridine complexes, this structure demonstrates that manganese(II) is no more prone to undergo low symmetry distortions than systems with ligand field stabilization energy contributions. PMID- 19129591 TI - A double-layered zinc(II) coordination polymer with the ligand 3,5 bis(carboxylatomethoxy)benzoate. AB - In the title compound, poly[hexaaquabis[mu(4)-3,5 bis(carboxylatomethoxy)benzoato]trizinc(II)], [Zn(3)(C(11)H(7)O(8))(2)(H(2)O)(6)](n), there are two crystallographically distinct Zn(II) cations which are bridged by polycarboxylate ligands in a mu(4) bridging mode. A pair of ligands bridges adjacent Zn atoms to give centrosymmetric dimetal building blocks which act as four-connected nodes to be further interlinked into a two-dimensional double-layered framework with (4,4) topology. Other Zn atoms, lying on inversion centres, occupy the cavities of this topological structure. This submission shows a versatile polycarboxylate ligand with rigid and flexible functional groups, the co-operation and complementarity of which would meet the coordination requirements of a variety of topological structures. PMID- 19129592 TI - (Di-tert-butylmethylphosphane)(eta2-di-tert butylphosphanylphosphinidene)(triphenylphosphane)platinum(0). AB - The crystal structure of the title compound, [Pt(C(8)H(18)P(2))(C(9)H(21)P)(C(18)H(15)P)] or [(Ph(3)P)((t)Bu(2)PMe)Pt(eta(2) (t)Bu(2)PP)], contains four molecules in the asymmetric unit with slightly different conformations. The P-P distances in the (t)Bu(2)PP ligands are similar for all four molecules [2.0661 (13)-2.0678 (13) A] and indicate a multiple character of the P-P bond in the (t)Bu(2)PP ligand. Molecules of the asymmetric unit can be assembled into a tetrahedron that fulfils the requirements for a rhombic disphenoid. The coordination of the Pt atom in all four molecules is square planar, with r.m.s. deviations from the PtP(4) planes in the range 0.03 0.05 A. All planes of the PtP(4) groups are approximately parallel to the ab plane of the unit cell. The structure represents an unusual unsymmetrical platinum phosphinidene derivative. PMID- 19129593 TI - Two isomorphous cobalt(II) complexes: poly[[diaqua-mu-2,5-dicarboxybenzene-1,4 dicarboxylato-mu-1,2-di-4-pyridylethene-cobalt(II)] 1,2-di-4-pyridylethene solvate] and the 1,2-di-4-pyridylethane analogue. AB - The two isomorphous title structures, formulated as {[Co(C(10)H(4)O(8))(C(12)H(10)N(2))(H(2)O)(2)].C(12)H(10)N(2)}(n), (I), and {[Co(C(10)H(4)O(8))(C(12)H(12)N(2))(H(2)O)(2)].C(12)H(12)N(2)}(n), (II), respectively, are reported. They crystallize in the space group P?overline{1} with only one formula unit in the asymmetric unit, so that the organic ligands lie about inversion centres and the Co atom lies on an inversion centre. The Co atoms are octahedrally coordinated by a carboxylate O atom from 2,5 dicarboxybenzene-1,4-dicarboxylate (H(2)btc), one N atom from 1,2-di-4 pyridylethene (L) in (I) or from 1,2-di-4-pyridylethane (L) in (II), and one coordinated water molecule, plus their inversion-related species. This particular coordination results in a two-dimensional array, with an elemental unit in the shape of a parallelogram having the Co(II) cations at the corners, linked in one direction by L bridges and in the opposite direction by H(2)btc groups. The L solvent molecules act as pillars between parallel planes, linking them by strong hydrogen bonds where the H atoms lie midway between the formal donor/acceptor atoms in a ;shared' mode. Comparison is made with structures presenting the same structural motif, strongly suggesting that the two-dimensional arrangement reported here might be a very stable robust building block for molecular engineering purposes. PMID- 19129594 TI - Two transition metal coordination polymers of the 7,7,8,8 tetracyanoquinodimethane dianion (TCNQ2-). AB - Each of the two novel title transition metal coordination polymers, namely catena poly[[bis{[tris(2-pyridylmethyl)amine]cobalt(II)}-mu(4)-7,7,8,8 tetracyanoquinodimethanide(2-)] bis[7,7,8,8-tetracyanoquinodimethanide(1-)] methanol disolvate], {[Co(2)(C(12)H(4)N(4))(C(18)H(18)N(4))(2)](C(12)H(4)N(4))(2).2CH(3)OH}(n), (I), and catena-poly[[[[tris(2-pyridylmethyl)amine]iron(II)]-mu(2)-7,7,8,8 tetracyanoquinodimethanide(2-)] methanol solvate], {[Fe(C(12)H(4)N(4))(C(18)H(18)N(4))].CH(3)OH}(n), (II), contains eta(4)-TPA and cis-bridging TCNQ(2-) ligands [TPA is tris(2-pyridylmethyl)amine and TCNQ is 7,7,8,8-tetracyanoquinodimethane], but the two compounds adopt entirely different structural motifs. Compound (I) consists of a ribbon coordination polymer featuring mu(4)-TCNQ(2-) radical anion ligands bridging four different octahedral Co(II) centers. Each formula unit of the polymer is flanked by two uncoordinated TCNQ(-) anions and two methanol solvent molecules. All three TCNQ anions have crystallographic inversion symmetry. In (II), the 2(1) symmetry operator generates a one-dimensional zigzag chain of octahedral Fe(II) centers with mu(2) TCNQ(2-) bridges. A methanol solvent molecule forms hydrogen bonds to one of the terminal N atoms of the bridging TCNQ(2-) dianion. To the best of our knowledge, these are the first examples of one-dimensional coordination polymers forming from cis coordination of two TCNQ ligands to octahedral metal centers. PMID- 19129595 TI - A new polymorph of poly[bis(mu2-perchlorato-kappa2O:O')(2,2':6',2''-terpyridine kappa3N,N',N'')lead(II)] with a greatly extended chain repeat distance. AB - In the title compound, [Pb(ClO(4))(2)(C(15)H(11)N(3))](n), two molecules occupy general positions while the third lies on a crystallographic twofold axis, giving a total of two and a half molecules per asymmetric unit. Each metal centre is coordinated equatorially by three 2,2':6',2''-terpyridine (terpy) N-donor atoms and axially by two perchlorate O-donor atoms. The distorted pentagonal bipyramidal geometry is completed by two equatorial O-donor atoms from two perchlorate anions which bridge to two different adjacent metal centres. The coordination about each metal centre is very similar to that seen at the unique Pb(II) centre in the previously published polymorph [Engelhardt, Harrowfield, Miyamae, Patrick, Skelton, Soudi & White (1996). Aust. J. Chem. 49, 1135-1146], but the new polymorph differs from it by the insertion on each side of an existing [bis(perchlorato)(terpy)lead(II)] molecule of two additional such units. Pairs of asymmetrically bridging perchlorate anions link irregularly spaced Pb(II) centres into undulating chains parallel to [201] which exhibit a repeat distance of 26.280 (4) A. The significance of this new polymorph lies in the fact that, while it is chemically identical to the known polymorph, it is structurally distinct from it. PMID- 19129596 TI - Probing the supramolecular interaction synthons of 1-benzofuran-2,3-dicarboxylic acid in its monoanionic form. AB - 1-Benzofuran-2,3-dicarboxylic acid (C(10)H(6)O(5)) is a dicarboxylic acid ligand which can readily engage in organometallic complexes with various metal ions. This ligand is characterized by an intramolecular hydrogen bond between the two carboxyl residues, and, as a monoanionic species, readily forms supramolecular adducts with different organic and inorganic cations. These are a 1:1 adduct with the dimethylammonium cation, namely dimethylammonium 3-carboxy-1-benzofuran-2 carboxylate, C(2)H(8)N(+).C(10)H(5)O(5)(-), (I), a 2:1 complex with Cu(2+) ions in which four neutral imidazole molecules also coordinate the metal atom, namely bis(3-carboxy-1-benzofuran-2-carboxylato-kappaO(3))tetrakis(1H-imidazole kappaN(3))copper(II), [Cu(C(10)H(5)O(5))(2)(C(3)H(4)N(2))(4)], (II), and a 4:1 adduct with [La(H(2)O)(7)](3+) ions, namely heptaaquabis(3-carboxy-1-benzofuran-2 carboxylato-kappaO(3))lanthanum 3-carboxy-1-benzofuran-2-carboxylate 1-benzofuran 2,3-dicarboxylic acid solvate tetrahydrate, [La(C(10)H(5)O(5))(2)(H(2)O)(7)](C(10)H(5)O(5)).C(10)H(6)O(5).4H(2)O, (III). In the crystal structure, complex (II) resides on inversion centres, while complex (III) resides on axes of twofold rotation. The crystal packing in all three structures reveals pi-pi stacking interactions between the planar aromatic benzofuran residues, as well as hydrogen bonding between the components. The significance of this study lies in the first crystallographic characterization of the title framework, which consistently exhibits the presence of an intramolecular hydrogen bond and a consequent monoanionic-only nature. It shows further that the anion can coordinate readily to metal cations as a ligand, as well as acting as a monovalent counter-ion. Finally, the aromaticity of the flat benzofuran residue provides an additional supramolecular synthon that directs and facilitates the crystal packing of compounds (I)-(III). PMID- 19129597 TI - A three-dimensional pyrazine-2,5-dicarboxylate Cd(II) coordination framework with new (4,4,4)-connected three-nodal topology. AB - Poly[(mu(4)-pyrazine-2,5-dicarboxylato)cadmium(II)], [Cd(C(6)H(2)N(2)O(4))](n) or [Cd(pzdc)](n) (pzdc is the pyrazine-2,5-dicarboxylate dianion), has been synthesized hydrothermally. The asymmetric unit consists of a Cd(II) atom and two independent halves of pzdc ligands that can be expanded via inversion through the centres of the ligands so that each ligand binds to four Cd(II) atoms with the same binding mode using six donor atoms. The Cd(II) centre is in a distorted octahedral coordination geometry with four O- and two N-atom donors from four pzdc ligands. The infinite linkage of the metal atoms and ligands forms a three dimensional framework with a rectangular channel which is so narrow that there is no measurable void space in the overall structure. This coordination polymer represents the first example of (4,4,4)-connected three-nodal framework. PMID- 19129598 TI - Poly[[di-mu-aqua-tetraaquadi-mu-hydroxido-bis(mu3-3-nitrophthalato)tricopper(II)] dihydrate]. AB - The novel title complex, {[Cu(3)(C(8)H(3)NO(6))(2)(OH)(2)(H(2)O)(6)].2H(2)O}(n), has a one-dimensional polymeric double chain structure where the three Cu atoms are linked by mu(2)-OH and mu(2)-H(2)O groups, and these trinuclear centres are bridged by two 3-nitrophthalate ligands. The asymmetric unit contains one and a half crystallographically independent Cu atoms (one lying on a centre of inversion), both coordinated by six O atoms and exhibiting distorted octahedral coordination geometries, but with different coordination environments. Each 3 nitrophthalate ligand connects to three Cu atoms through two O atoms of one carboxylate group and one O atom of the nitro group. The remaining carboxylate group is free and is involved in intrachain hydrogen bonds, reinforcing the chain linkage. PMID- 19129599 TI - Poly[[diaqua-mu-4,4'-bipyridine-dinitratodi-mu-L-tyrosinato-dicopper(II)]: a chiral two-dimensional coordination polymer. AB - The title compound, [Cu(2)(C(9)H(10)NO(3))(2)(NO(3))(2)(C(10)H(8)N(2))(H(2)O)(2)](n), contains Cu(II) atoms and L-tyrosinate (L-tyr) and 4,4'-bipyridine (4,4'-bipy) ligands in a 2:2:1 ratio. Each Cu atom is coordinated by one amino N atom and two carboxylate O atoms from two L-tyr ligands, one N atom from a 4,4'-bipy ligand, a monodentate nitrate ion and a water molecule in an elongated octahedral geometry. Adjacent Cu atoms are bridged by the bidentate carboxylate groups into a chain. These chains are further linked by the bridging 4,4'-bipy ligands, forming an undulated chiral two-dimensional sheet. O-H...O and N-H...O hydrogen bonds connect the sheets in the [100] direction. This study offers useful information for the engineering of chiral coordination polymers with amino acids and 4,4'-bipy ligands by considering the ratios of the metal ion and organic components. PMID- 19129600 TI - 1,1'-Dimethoxy-3,3'-dimethyl-3,3'-(hexane-1,6-diyl)bis(triazen-2-ium-2-olate): a nitric oxide donor. AB - The title compound, C(10)H(24)N(6)O(4), is the most stable type of nitric oxide (NO) donor among the broad category of discrete N-diazeniumdiolates (NO adducts of nucleophilic small molecule amines). Sitting astride a crystallographic inversion center, the molecule contains a symmetric dimethylhexane-1,6-diamine structure bearing two planar O(2)-methylated N-diazeniumdiolate functional groups [N(O)=NOMe]. These two groups are parallel to each other and have the potential to release four molecules of NO. The methylated diazeniumdiolate substituent removes the negative charge from the typical N(O)=NO(-) group, thereby increasing the stability of the diazeniumdiolate structure. The crystal was nonmerohedrally twinned by a 180 degrees rotation about the real [101] axis. This is the first N based bis-diazeniumdiolate compound with a flexible aliphatic main unit to have its structure analyzed and this work demonstrates the utility of stabilizing the N-diazeniumdiolate functional group by methylation. PMID- 19129601 TI - 2-Amino-4-chloro-5-formyl-6-[methyl(2-methylphenyl)amino]pyrimidine and 2-amino-4 chloro-5-formyl-6-[(2-methoxyphenyl)methylamino]pyrimidine are isostructural and form hydrogen-bonded sheets of R2(2)8 and R6(6)32 rings. AB - 2-Amino-4-chloro-5-formyl-6-[methyl(2-methylphenyl)amino]pyrimidine, C(13)H(13)ClN(4)O, (I), and 2-amino-4-chloro-5-formyl-6-[(2 methoxyphenyl)methylamino]pyrimidine, C(13)H(13)ClN(4)O(2), (II), are isostructural and essentially isomorphous. Although the pyrimidine rings in each compound are planar, the ring-substituent atoms show significant displacements from this plane, and the bond distances provide evidence for polarization of the electronic structures. In each compound, a combination of N-H...N and N-H...O hydrogen bonds links the molecules into sheets built from centrosymmetric R(2)(2)(8) and R(6)(6)(32) rings. The significance of this study lies in its observation of the isostructural nature of (I) and (II), and in the comparison of their crystal and molecular structures with those of analogous compounds. PMID- 19129602 TI - (2S,3R,4S,5R)-Diethyl 2-(10,10-dimethyl-3,3-dioxo-3lambda(6)-thia-4 azatricyclo[5.2.1.0(1,5)]decan-4-ylcarbonyl)-5-phenylpyrrolidine-3,4 dicarboxylate: a novel isomorphous-by-addition compound. AB - The title compound, C(27)H(36)N(2)O(7)S, (I), is isomorphous by addition with the dimethyl ester analogue [Garner, Dogan, Youngs, Kennedy, Protasiewicz & Zaniewski (2001). Tetrahedron, 57, 71-85], (II), by replacing two methyl ester H atoms with two methyl groups. With the exception of the conformation of one of the ester groups, the molecules are almost superimposable. Likewise, apart from a slightly larger c axis in (I), few differences in the cell packing of (I) and (II) are found, with both dominated by the same C-H...O hydrogen bonds. Full synthetic and spectroscopic details of (I) are given. The molecular synthesis is important as an example of chiral auxiliary-assisted 1,3-dipolar cycloaddition of an azomethine ylid. PMID- 19129603 TI - 2,3-Dihydro-1,3-benzothiazol-2-iminium hydrogen oxydiacetate: a combined structural and theoretical study. AB - In the title compound, C(7)H(7)N(2)S(+).C(4)H(5)O(5)(-), the ions are connected by N-H...O hydrogen bonds. The hydrogen oxydiacetate residues are linked together by O-H...O hydrogen bonds disordered about centres of inversion into hydrogen bonded ribbon layers crosslinked by weak C-H...O and stacking interactions. The cation exists mainly in the 2,3-dihydro-1,3-benzothiazol-2-iminium form, with a small participation of the 2-aminobenzothiazolium form, based on the structural data and quantum mechanical calculations. This study provides structural insights relevant to the biochemical activity of benzothiazole molecules. PMID- 19129604 TI - Two polymorphs of morpholin-4-ium 2-(5-methyl-1H-1,2,4-triazol-3 ylsulfanyl)acetate. AB - Two polymorphs of the title organic salt (a very effective medicinal preparation with the commercial name thiotriazoline), C(4)H(10)NO(+).C(5)H(6)N(3)O(2)S(-), were obtained. The cations and anions are connected by hydrogen bonds and extend into two-dimensional networks. The main packing motifs are an R(4)(4)(12) cluster in the monoclinic form and a chain in the orthorhombic form. PMID- 19129605 TI - Two similar dibenzo cyclic ethers with dissimilar conformations. AB - Two dibenzo cyclic ether compounds, 6,12-dibromodibenzo[d,i]-1,2,3,6,7,8 hexahydro-1,3-dioxecin (systematic name: 8,16-dibromo-2,4 dioxatricyclo[12.4.0.0(5,10)]octadeca-5,7,9,14,16,18-hexaene), C(16)H(14)Br(2)O(2), (II), and 8,14-dibromodibenzo[f,k]-1,5-dioxa 1,2,3,4,5,8,9,10-octahydrocyclododecene (systematic name: 7,19-dibromo-11,15 dioxatricyclo[14.4.0.0(5,10)]icosa-5,7,9,16,18,20-hexaene), C(18)H(18)Br(2)O(2), (III), were prepared as scaffolding for phosphate-anion receptors. In both compounds, the two aromatic rings are linked by three methylene units ortho to the oxygen substituent of each ring. The only difference between the two compounds is the number of methylene units linking the two ether O atoms. The dibenzo cyclic ether with an ether linkage of one methylene unit adopts a chair like conformation, where the two aromatic rings are parallel to each other. On the other hand, the dibenzo cyclic ether with an oxygen linkage of three methylene units adopts a bowl-like conformation. The latter scaffold configuration is the only structure of the two that would allow for the placement of convergent functional groups necessary for the establishment of an anion selective binding pocket. PMID- 19129606 TI - Hydrogen-bonded assemblies of 5,10,15,20-tetrakis(4-hydroxyphenyl)porphyrin with dimethylformamide, dimethylacetamide and water. AB - The title free base porphyrin compound forms hydrogen-bonded adducts with N,N dimethylformamide, C(44)H(30)N(4)O(4).4C(3)H(7)NO, (I), a mixture of N,N dimethylformamide and water, C(44)H(30)N(4)O(4).4C(3)H(7)NO.H(2)O, (II), and a mixture of N,N-dimethylacetamide and water, C(44)H(30)N(4)O(4).6C(3)H(7)NO.2H(2)O, (III). Total solvation of the four hydroxy functions of the porphyrin molecules characterizes all three compounds, thus preventing its supramolecular association into extended network architectures. In (I), the asymmetric unit consist of two five-component adduct species, while in (III), the nine-component entities reside on centres of inversion. This report provides the first structural characterizations of the free base tetra(hydroxyphenyl)porphyrin. It also demonstrates that the presence of strong Lewis bases, such as dimethylformamide or dimethylacetamide, in the crystallization mixture prevents direct supramolecular networking of the porphyrin ligands via O-H...O-H hydrogen bonds, due to their competing O H...N(base) interaction with the hydroxy functions. The crystal packing of compounds (I)-(III) resembles that of other hydrogen-bonding-assisted tetraarylporphyrin clathrates. PMID- 19129607 TI - The zwitterion of 4-nitro-2-{(E)-[2-(piperidin-1-yl)ethyl]iminomethyl}phenol. AB - The title Schiff base compound, 4-nitro-1-oxo-2-{(E)-[2-(piperidin-1 yl)ethyl]iminiomethyl}cyclohexadienide, C(14)H(19)N(3)O(3), exists as a zwitterion, with the H atom of the phenol group being transferred to the imine N atom. The C=O, C(Ar)-C(Ar) and C-N bond lengths are in agreement with the oxocyclohexadienide-iminium zwitterionic form. The iminium H atom is engaged in a strong intramolecular hydrogen bond with the O atom of the keto group (N(+) H...O) to form an S(6) motif. Soft C-H...O interactions in the ac plane lead to the development of hydrogen-bonded tapes, which are pi-stacked through the oxocyclohexadienide ring and iminium group. The significance of this study is in providing crystallographic evidence, supported by NMR and IR data, of the predominance of the oxocyclohexadienide-iminium zwitterion form over the noncharged canonical form in the title Schiff base. PMID- 19129608 TI - Early use of ERCP in acute biliary pancreatitis with(out) jaundice: an unjaundiced view. PMID- 19129609 TI - Icam-1 and acute pancreatitis complicated by acute lung injury. AB - One of the most common complications of acute pancreatitis is acute lung injury, during which intercellular adhesion molecule-1 (ICAM-1) plays an important role by participating in leukocyte adhesion and activation as well as by inducing the "cascade effect" of inflammatory mediators, pulmonary microcirculation dysfunction and even acute respiratory distress syndrome, multiple organ failure or death. Although it is generally believed that the modulatory mechanism of ICAM 1 during this process is associated with the activation of nuclear transcription factor kappa B which is mediated by IL-1, IL-6, IL-18 and oxygen free radical, etc., further studies are still required to clarify it. Since the upregulation of ICAM-1 expression in the lung during acute lung injury is one of main pathogeneses, the early detection of the ICAM-1 expression level may contribute to the prevention and treatment of acute lung injury. Moreover, reducing pulmonary ICAM-1 expression levels through treatment with anti-ICAM-1 monoclonal antibody (aICAM-1) and antagonists of the neurokinin 1 receptor, etc., should have a positive effect on protecting the lungs during acute pancreatitis. This review aims to further clarify the relationship between ICAM-1 and acute pancreatitis complicated by acute lung injury, and therefore provides a theoretical basis for the formulation of corresponding therapeutic measures in clinical practice for acute pancreatitis. PMID- 19129610 TI - Administration of anti-Reg I and anti-PAPII antibodies worsens pancreatitis. AB - CONTEXT: The regeneration protein family (Reg), which includes Reg I and PAPII, is expressed in pancreas acinar cells, and increases in acute pancreatitis. We have demonstrated that Reg gene knockdown worsens severity of acute pancreatitis in the rat and hypothesize that the proteins offer a protective effect in this disease. OBJECTIVE: We investigated the ability of anti-Reg and anti-PAP antibody to neutralize pancreatic Reg protein and affect pancreatitis severity. INTERVENTION: Pancreatitis was induced in rats by retrograde ductal injection of 4% sodium taurocholate. ANIMALS: Eighty-four rats: 48 with induced pancreatitis, 30 sham operated, and 6 normal animals. SETTING: Intraductal anti-Reg I and/or anti-PAPII antibody was administered at induced pancreatitis and sham operated subgroups of 6 rats each. MAIN OUTCOME MEASURE: Serum and pancreata were harvested 24 and/or 48 hours later and assessed for pancreatitis severity by pancreatic wet weight, serum C-reactive protein (CRP), amylase, PAPII levels, and histopathology. RESULTS: Animals induced with pancreatitis with administration of anti-Reg/PAP antibodies had significantly higher wet weights compared with taurocholate and histopathological analysis revealed that anti-Reg/PAP treated animals had worse tissue inflammation and necrosis compared with controls. Serum CRP, amylase, and Reg levels did not significantly differ between experimental and sham control groups. CONCLUSIONS: Administration of anti-Reg/PAP antibody worsened taurocholate-induced organ specific pancreatitis. These data suggest that the Reg family of proteins is protective in acute pancreatitis. PMID- 19129611 TI - Relationship of postoperative complications from preoperative biliary stents after pancreaticoduodenectomy. A new cohort analysis and meta-analysis of modern studies. AB - CONTEXT: Debate still continues as to the effects of preoperative biliary stents on postoperative complications after pancreaticoduodenectomy. Some studies have documented increased wound infection rates, while others have not. The importance of this issue rests on whether these postoperative complications are detrimental enough to not recommend preoperative chemoradiation in the treatment of pancreatic cancer. OBJECTIVE: This study is in two parts: 1) a retrospective review of patients who underwent pancreaticoduodenectomy at Henry Ford Hospital; and 2) a meta-analysis of published studies on the effects of preoperative biliary stents. METHODS: In the retrospective portion, all patients who underwent pancreaticoduodenectomy from January 1st, 1997 through December 31st, 2006 were included in the study. MAIN OUTCOME MEASURES: Data gathered included gender, age, pathologic diagnosis, use of preoperative biliary stent (either ERCP or PTC), all postoperative complications, and in-hospital mortality. In the meta-analysis portion, all studies published from 1990 with either a randomized or quasi randomized allocation of patients were included. Endpoints analysis were peri operative mortality, wound infection rate, intra-abdominal abscess rate, and overall morbidity rate. RESULTS: In the retrospective portion, 181 patients were studied, with 123 (68.0%) of these having preoperative biliary stents. Patients with and without stents had no significant difference in wound infection rate (19.5% vs. 17.2%, respectively), intra-abdominal abscess rate (16.3% vs. 22.4%), any postoperative complication (50.4% vs. 51.7%) and in-hospital death (2.4% vs. 1.7%). Fifteen studies were included in the meta-analysis. There was variation in both the definitions of complications as well as the incidence of all postoperative endpoints among the studies. For peri-operative mortality and wound infection rate, the relative difference favored the no stent group by 0.5% (95% confidence interval: -0.4% to 1.4%) and 5.8% (95% confidence interval: 3.6% to 8.0%), respectively. For intra-abdominal abscess and overall morbidity rate, the relative difference favored the stent group by 2.0% (95% confidence interval: 0.3% to 4.3%) and 0.06% (95% confidence interval -3.8% to 3.9%), respectively. CONCLUSION: Although the use of a preoperative biliary stent increases the postoperative wound infection rate by about 5%, there is no overwhelming evidence that it either promotes or protects from the other complications. As there was variation in the definitions used in these studies, a more uniformed system of complication reporting is required. PMID- 19129612 TI - Variance in elective surgery for chronic pancreatitis. AB - CONTEXT: Evidence to guide selection of optimal surgical treatment for patients with painful chronic pancreatitis is limited. Baseline assessment data are limited and thus patients in different centres may be presenting at different stages of their illness. OBJECTIVE: This study undertakes a systematic overview of reports of elective surgical intervention in chronic pancreatitis with particular reference to reporting of quality of life and baseline assessment and relation between disease and type of procedure. METHODS: A computerised search of the PubMed, Embase and Cochrane databases was undertaken for the period January 1997 to March 2007 yielding 46 manuscripts providing data on 4,626 patients undergoing elective surgery for chronic pancreatitis. The median number of patients per study was 71 (range: 4-484). The median period for recruitment of patients was 10 years (range: 2-36 years). RESULTS: An externally validated quality of life questionnaire is reported in 8 (17.4%) of 46 manuscripts covering 441 (9.5%) of 4,626 patients. Formal comparison of pre-operative and post operative pain scores was provided in 15 (32.6%) of manuscripts. Only seven (15.2%) reports provide a formal rationale or indication for selection of the type of elective surgical procedure for a stated disease variant and these papers cover 481 (10.4%) patients. CONCLUSION: In conclusion, this study demonstrates that there is a lack of standardization between units of the criteria for operative intervention in painful chronic pancreatitis. At a minimum, formal quality of life testing using a validated system should be undertaken in all patients prior to elective surgery for painful chronic pancreatitis. PMID- 19129613 TI - Focal or diffuse "fullness" of the pancreas on CT. Usually benign, but EUS plus/minus FNA is warranted to identify malignancy. AB - CONTEXT: The role of EUS to evaluate subtle radiographic abnormalities of the pancreas is not well defined. OBJECTIVE: To assess the yield of EUS+/-FNA for focal or diffuse pancreatic enlargement/fullness seen on abdominal CT scan in the absence of discrete mass lesions. DESIGN: Retrospective database review. SETTING: Tertiary referral center. PATIENTS AND INTERVENTIONS: Six hundred and 91 pancreatic EUS exams were reviewed. Sixty-nine met inclusion criteria of having been performed for focal enlargement or fullness of the pancreas. Known chronic pancreatitis, pancreatic calcifications, acute pancreatitis, discrete mass on imaging, pancreatic duct dilation (greater than 4 mm) and obstructive jaundice were excluded. MAIN OUTCOME MEASUREMENT: Rate of malignancy found by EUS+/-FNA. RESULTS: FNA was performed in 19/69 (27.5%) with 4 new diagnoses of pancreatic adenocarcinoma, one metastatic renal cell carcinoma, one metastatic colon cancer, one chronic pancreatitis and 12 benign results. Eight patients had discrete mass lesions on EUS; two were cystic. All malignant diagnoses had a discrete solid mass on EUS. CONCLUSIONS: Pancreatic enlargement/fullness is often a benign finding related to anatomic variation, but was related to malignancy in 8.7% of our patients (6/69). EUS should be strongly considered as the next step in the evaluation of patients with focal enlargement of the pancreas when clinical suspicion of malignancy exists. PMID- 19129615 TI - Hemosuccus pancreaticus due to primary splenic artery aneurysm: a diagnostic and therapeutic challenge. AB - CONTEXT: Hemosuccus pancreaticus is a rare cause of upper chronic and intermittent gastrointestinal hemorrhage which cannot be easily detected by endoscopy. It is usually due to the rupture of a visceral aneurysm into the main pancreatic duct; splenic artery pseudoaneurysm associated with chronic pancreatitis represents the leading cause of this condition. The diagnosis is based on direct visualization of the hemorrhage through the main pancreatic duct at angiography. Given its rarity, difficulties in determining the source of bleeding can result in delayed treatment. CASE REPORT: We present a rare case of true splenic artery aneurysm fistulized in the main pancreatic duct and misdiagnosed as a bleeding pancreatic pseudocyst on preoperative examination which included CT and MRCP. CONCLUSIONS: Our experience confirms that the diagnosis of bleeding from the main pancreatic duct is very difficult. It requires careful and repeated evaluation by a team of specialists. PMID- 19129614 TI - Management guidelines for gallstone pancreatitis. Are the targets achievable? AB - CONTEXT: Current management of gallstone pancreatitis in a university hospital. OBJECTIVE: Comparison of current management of gallstone pancreatitis with recommendations in national guidelines. SETTING: Tertiary care centre in Scotland. SUBJECTS: One-hundred consecutive patients admitted with gallstone pancreatitis. METHODS: All patients that presented with gallstone pancreatitis over a 4-year period were audited retrospectively. Data were collated for radiological diagnosis within 48 hours, ERCP within 72 hours, CT at 6-10 days, and use of high-dependency or intensive therapy units in severe gallstone pancreatitis, and definitive treatment of gallstone pancreatitis within 2 weeks as recommended in national guidelines. RESULTS: Forty-six patients had severe gallstone pancreatitis and 54 patients mild pancreatitis. Etiology was established within 48 hours in 92 patients. Six (13.0%) out of the patients with severe gallstone pancreatitis were managed in a high dependency unit. Fifteen (32.6%) patients with severe gallstone pancreatitis underwent CT within 6-10 days of admission. Four (8.7%) of the 46 patients with severe gallstone pancreatitis had urgent ERCP (less than 72 hours). Overall 22/100 patients unsuitable for surgery underwent endoscopic sphincterotomy as definitive treatment. Seventy eight patients had surgery, with 40 (51.3%) of these patients undergoing an index admission cholecystectomy, and 38 (48.7%) patients were discharged for interval cholecystectomy. Overall 81 patients with gallstone pancreatitis had definitive therapy during the index to same admission (cholecystectomy or sphincterotomy). Two (5.3%) patients were readmitted whilst awaiting interval cholecystectomy: one with acute cholecystitis and one with acute pancreatitis. There were no mortalities in this cohort. CONCLUSION: This study has highlighted difficulties in implementation of national guidelines, as the use of critical care, timing of ERCP and CT, and definitive treatment prior to discharge did not concur with national targets for gallstone pancreatitis. PMID- 19129616 TI - Ischemic acute necrotizing pancreatitis in a marathon runner. AB - CONTEXT: Acute pancreatitis due to pancreatic ischemia is a rare condition. CASE REPORT: In this case report we describe a 57-year-old male who developed an acute necrotizing pancreatitis after running a marathon and visiting a sauna the same evening, with an inadequate fluid and food consumption during both events. CONCLUSIONS: Pancreatic ischemia imposed by mechanical and physical stress and dehydration can induce the development of acute pancreatitis. Separately, these factors are rare causes of ischemic acute pancreatitis. But when combined, as in this particular case, the risk of an acute necrotizing pancreatitis cannot be neglected. PMID- 19129617 TI - Diffuse pancreatic serous cystadenoma associated with neuroendocrine carcinoma: a case report and review of literature. AB - CONTEXT: Diffuse serous cystadenomas of the pancreas are extremely rare, with only 8 cases reported previously, and have been associated with neuroendocrine tumors in only two patients. Some have been seen in von Hippel-Lindau disease. The management of these tumors poses a challenge due to their rarity and uncertain malignant potential. We report a case of diffuse serous cystadenoma associated with neuroendocrine carcinoma in a 35-year-old woman. CASE REPORT: A 35-year-old woman with mild abdominal pain was diagnosed as having a cystic pancreatic mass on ultrasonography. On contrast-enhanced CT scan, MRI and MRCP imaging, a spongy lesion was found to replace the entire pancreas, and was diagnosed as diffuse serous cystadenoma. Serum biochemistry for amylase, lipase, CA 19-9 and CEA was normal. Screening for retinal and CNS lesions was also unremarkable. A total pancreatectomy was performed, and the patient recovered well. Histopathological examination of the specimen revealed microcysts and macrocysts replacing the entire pancreas, the largest being 3.5 cm. The cysts were lined with a single layer of cuboidal to flattened cells. An endocrine tumor abutting the cystic component was found, having neoplastic cells in a trabecular pattern. Metastasis of the neuroendocrine component was seen in the adherent lymph nodes. A diagnosis of diffuse serous cystadenoma associated with neuroendocrine carcinoma was made. CONCLUSION: Diffuse serous cystadenomas of the pancreas are extremely rare tumors. In young patients, they may harbour associated malignancy, and may be the first presentation of von Hippel-Lindau disease. Aggressive surgical resection with long-term follow-up may be worthwhile in this group of patients. PMID- 19129618 TI - Pancreatic arteriovenous malformation: a case report of hemodynamic and three dimensional morphological analysis using multi-detector row computed tomography and post-processing methods. AB - CONTEXT: Although rare, a pancreatic arteriovenous malformation can have serious consequences. A diagnosis of arteriovenous malformation requires evidence of aberrant communication between the arterial and the venous systems. This report describes a case where the use of multi-detector row CT and specific post processing methods provided a diagnosis of arteriovenous malformation. This minimally invasive diagnostic approach resulted in a clear, precise and comprehensive visual representation of the pancreatic arteriovenous malformation. CASE REPORT: A 60-year-old man with right hypochondriac pain presented with a mass in the head of the pancreas. The hypochondriac pain resolved spontaneously and physical examination revealed no abnormal findings. A multi-detector row CT study was performed. The data obtained in the arterial phase demonstrated a high contrast mass in the head of the pancreas and early enhancement of the portal vein. A maximum intensity projection method clarified the aberrant vascular communication. Changes in Hounsfield numbers were observed using a multi-planar reformation method. A volume-rendering method was used to create a 3D model which demonstrated the spatial relationship between the aberrant vascular communication and the surrounding tissue. An annual follow-up study using this technique showed no significant alteration. CONCLUSIONS: Multi-detector row CT with specific post processing methods is a useful diagnostic tool for pancreatic arteriovenous malformation. PMID- 19129619 TI - Isolated tuberculosis of the pancreas: a case report. AB - CONTEXT: Pancreatic tuberculosis is a rare entity. Only a few cases have been reported in the medical literature. We hereby describe a case of pancreatic tuberculosis in an immunocompromized individual. CASE REPORT: A fifty-year-old African-American gentleman with history of HIV non-compliant on anti-retroviral therapy presented with epigastric pain for five weeks duration. CT scan of abdomen showed large necrotic node on the posterior aspect of the head of pancreas and multiple cystic masses adjacent to the pancreas. Acid fast bacilli were found on staining of CT guided biopsy of the node. Cultures grew Mycobacterium tuberculosis. Anti-tubercular therapy was initiated and resulted in gradual resolution of symptoms. CONCLUSION: Pancreatic tuberculosis is rare and is frequently confused with pancreatic cancer on clinical presentation as well as on imaging studies. Since it is a curable disease, accurate diagnosis is paramount CT or ultrasound guided biopsy is cornerstone of diagnosis. Endoscopic ultrasound is now increasingly being used for obtaining tissue for diagnosis. Anti-tubercular therapy is curative in majority of the cases. PMID- 19129620 TI - The relationship of body underweight to complication risks in acute pancreatitis. PMID- 19129621 TI - Depression and pancreatic cancer: a poorly understood link. PMID- 19129623 TI - Identification of a gene cluster of polyether antibiotic lasalocid from Streptomyces lasaliensis. AB - Elucidation of enzymatic polyether formation is a long-standing controversial issue in organic chemistry. To address this intriguing issue, identifying the actual substrate for epoxidation and sequential cyclization is essential. We selected the representative polyether ionophore, lasalocid, which has been proposed to undergo no modification at the late stage of biosynthesis. Cloning and a sequence analysis revealed seven polyketide synthase (PKS) genes, epoxidase and epoxide hydrolase genes for sequential ether formation, and several putative genes for supplying ethylmalonyl-CoA. Based on bioinformatic data, we propose the lasalocid biosynthetic pathway which involves characteristic aromatic ring formation and sequential cyclic ether formation. The finding of a thioesterase domain at the C-terminal of the seventh PKS indicates that intriguing oxidative cascade cyclization would occur after cleavage of the polyketide intermediate from PKS. Based on this observation, we have recently reported the enzymatic transformation of a bisepoxide intermediate to lasalocid with the recombinant epoxide hydrolase, Lsd19. PMID- 19129624 TI - Surugapyrroles A and B, two new N-hydroxypyrroles, as DPPH radical-scavengers from Streptomyces sp. USF-6280 strain. AB - Two new N-hydroxypyrroles, surugapyrroles A and B, were isolated as DPPH radical scavengers from a culture broth of Streptomyces sp. USF-6280 strain. The structures of the two compounds were elucidated by spectroscopic evidences and derivatization. Their DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging activity was as high as that of butylhydroxytoluene (BHT): surugapyrrole A, ED(50) 50.3 microM; surugapyrrole B, ED(50) 75.8 microM. PMID- 19129625 TI - New sesquiterpenoid from the mushroom Sparassis crispa. AB - A new sesquiterpenoid (1) was isolated from the mushroom, Sparassis crispa, along with a known sesquiterpenoid (2). The structure of 1 was determined as (3R(*), 3aS(*), 4S(*), 8aR(*))-3-(1'-hydroxy-1'-methylethyl)-5,8a-dimethyldecahydroazulen 4-ol by the combination of NMR and ESI-MS analyses. PMID- 19129626 TI - Purification and characterization of the first archaeal glutamate decarboxylase from Pyrococcus horikoshii. AB - Glutamate decarboxylase (GAD) from the archaeon Pyrococcus horikoshii was successfully expressed and purified, with the aim of developing a hyperthermostable GAD for industrial applications. Its biochemical properties were different from those reported for other GADs. The enzyme had broad substrate specificity, and its optimum pH and temperature were pH 8.0 and > 97 degrees C. PMID- 19129627 TI - Sleeping Beauty-mediated suicide gene therapy of hepatocellular carcinoma. AB - The aim of this study was to use gene therapy via the Sleeping Beauty (SB) system to increase telomerase promoter activity to target hepatocellular carcinoma (HCC). In previous studies, we identified selective and increased expression of luciferase and suicide genes controlled by the hTERT (human telomerase reverse transcriptase) promoter and the SV40 enhancer in telomerase-positive cancer cell lines. Because telomerase is activated in about 80% of HCCs, it is likely that increasing the activity of the telomerase promoter with a suicide gene will effectively eradicate HCCs. We found that the telomerase promoter mediated SB system can efficiently insert transgene into HCC genomes. Also, telomerase promoter activity was increased using a SB vector expressing suicide gene HSV-TK (herpes simplex virus thymidine kinase) controlled by the hTERT promoter and a SV40 enhancer for the induction of telomerase-positive cancer-specific cell death. HCC cell lines transfected with pT.hTp.HSV-tk.Con with active helper plasmid and ganciclovir (GCV) significantly inhibited cancer cell growth. These results indicate that Sleeping Beauty transposon mediated suicide gene expression can be used in HCC-targeted cancer gene therapy. PMID- 19129628 TI - Predominant occurrence of truncated complex type N-glycans among glycoproteins in mature red tomato. AB - In this study, we analyzed the structures of complex type N-glycans linked to glycoproteins in mature red tomato to determine the relative ratio of high mannose type structure and complex type structure. Structural analysis of pyridylaminated N-glycans revealed that the truncated plant complex type structure accounted for about 70% of total conjugated N-glycans, while the high mannose type structure accounted for about 22%. PMID- 19129629 TI - Synthesis of novel conjugates of tetraoxane endoperoxide with bis(quaternary ammonium salts). AB - Novel water-soluble conjugates of 1,2,4,5-tetraoxane bis(quaternary ammonium salts) were synthesized in a relatively stable crystalline form via four steps starting from methyltrioxorhenium-catalyzed endo-peroxidation of ethyl 4 oxocyclohexanecarboxylate with hydrogen peroxide in hexafluoro-2-propanol. The assay for the in vitro toxicity of water-soluble tetraoxanes 5a-5d to malaria parasites indicate that they were inactive against the Plasmodium falciparum FCR 3 strain. PMID- 19129630 TI - An approach to strain improvement and enhanced production of clavulanic acid in Streptomyces clavuligerus. AB - To develop a strategy for improved production of clavulanic acid (CA), we investigated the effect of using oils on cell growth and CA production during the fermentation of Streptomyces clavuligerus NRRL 3585. In this analysis, triolein, whose fatty acid is oleic acid only, was the best oil source for CA production, but free fatty acids generated from the hydrolysis of oils in a culture broth negatively impacted CA production and cell growth. Hence, we screened for mutants that were resistant to high concentrations of oleic acid. From this screen we identified a mutant S. clavuligerus, OL13, that had a minimum inhibitory concentration (MIC) to oleic acid of 2.1 g/l, much higher than that of S. clavuligerus NRRL 3585, at 0.4 g/l. Not only was cell growth improved, but maximum CA production, at 1,950 mg/l, was approximately 2.0-fold higher than that of the parent strain. PMID- 19129631 TI - Identification and application of novel autonomously replicating sequences (ARSs) for promoter-cloning and co-transformation in Candida utilis. AB - In order to develop practical recombinant DNA techniques in the industrially important yeast Candida utilis, at least six plasmids harboring autonomously replicating sequences (ARSs) were isolated from a C. utilis genomic library. Two ARSs were subjected to detailed analysis. Sequences of 1.9 and 1.8 kb were found to be necessary to exert ARS activity in a plasmid as assessed by transformation efficiency and mitotic stability. Both fragments were found to be rich in AT content (69.5% and 70.8% respectively), and to contain an 11-bp ARS consensus sequences (10 and 13 motifs with one base difference respectively). Using the ARS containing plasmid as a promoter-cloning vector, several DNA fragments having promoter activities were cloned and characterized. Co-transformation of C. utilis with an integrating DNA fragment and a replicating plasmid yielded plasmid-free transformants harboring the fragment integrated into the C. utilis genome. PMID- 19129632 TI - WeGAS: a web-based microbial genome annotation system. AB - We have developed WeGAS, a Web based microbial Genome Annotation System, which provides features that include gene prediction, homology search, promoter/motif analysis, genome browsing, gene ontology analysis based on the COGs and GO, and metabolic pathway analysis with web-based interfaces. Most raw data and intermediate data from genome projects can be managed with the WeGAS database system, and analysis results, including information on each gene and final genome maps, are provided by its visualization modules. Especially, a pie-view browser displaying circular maps of contigs and a COG-GO combination browser are very helpful for an overview of projects. Major public microbial genome databases can be imported, searched, and browsed through the WeGAS modules. WeGAS is freely accessible via web site http://ns.smallsoft.co.kr:8051. PMID- 19129633 TI - KIBRA Co-localizes with protein kinase Mzeta (PKMzeta) in the mouse hippocampus. AB - KIBRA is a WW domain-containing protein that can bind to protein kinase Czeta (PKCzeta). The SNP of the ninth intron of the KIBRA gene is associated with human episodic memory performance. Protein kinase Mzeta (PKMzeta), a brain-specific variant of PKCzeta, plays important roles in memory formation. Here we examined the interaction of KIBRA and PKMzeta in the adult mouse brain. Immunoprecipitation using newly-raised anti-KIBRA antibody revealed the interaction between KIBRA and PKMzeta in the brain. KIBRA was co-localized with PKMzeta in a single cultured neuron. Distribution analysis by immunohistochemistry and in situ hybridization indicated that KIBRA was highly localized with PKMzeta in the hippocampal CA1, CA3, and dentate gyrus. These results suggest that KIBRA functions in memory performance via interaction with PKMzeta. PMID- 19129634 TI - alpha-mannosidase involved in turnover of plant complex type N-glycans in tomato (Lycopersicum esculentum) fruits. AB - In this study, we purified and characterized an alpha-mannosidase to homogeneity from mature red tomato fruits. Purified alpha-mannosidase (alpha-Man LE-1) gave two separate bands, of molecular masses of 70 kDa (L-subunit) and 47 kDa (S subunit), on SDS-PAGE under non-reducing and reducing conditions. On the other hand, the molecular weight was estimated to be 230 kDa by gel filtration, indicating that alpha-Man LE-1 functions in a tetrameric structure in plant cells. The N-terminal sequence of the L-subunit and the S-subunit were determined to be L-Y-M-V-Y-M-T-K-Q-G- and X-X-L-E-Q/K-S-F-S-Y-Y respectively. When pyridylaminated N-glycans were used as substrates, alpha-Man LE-1 showed optimum activity at about pH 6 and at 40 degrees C, and the activity was completely inhibited by both swainsonine and 1-deoxy-mannojirimycin. alpha-Man LE-1 hydrolyzed the alpha-mannosidic linkages from both high-mannose type and plant complex type N-glycan, but preferred a truncated plant complex type structure to high-mannose type N-glycans bearing alpha1-2 mannosyl residues. PMID- 19129635 TI - Inhibition of pathogen adhesion to host cells by polysaccharides from Panax ginseng. AB - PG-F2 and PG-HMW from Panax ginseng are pectin-type polysaccharides and PG-HMW might be an arabinogalactan. They demonstrated strong anti-adhesive activities against oral and skin pathogens to host cell lines in a dose-dependent manner from 0.1 to 2.0 mg/ml. While enzymatic hydrolysis caused complete loss of anti adhesive activities, partial hydrolysis produced oligosaccharides with anti adhesive properties. PG-F2 and PG-HMW might have a selective anti-adhesive effect against certain pathogenic bacteria without adverse effects on commensal bacteria. PMID- 19129636 TI - Solubilization, purification, and properties of membrane-bound D-glucono-delta lactone hydrolase from Gluconobacter oxydans. AB - Membrane-bound glucono-delta-lactonase (MGL) was purified to homogeneity from the membrane fraction of Gluconobacter oxydans IFO 3244. After solubilization with 1 M CaCl2, MGL was purified in the presence of Ca2+ and detergent. A single band corresponding to 60 kDa appeared in SDS-PAGE. The molecular weight of MGL was judged to be 120 k. Differently from cytoplasmic lactonases, MGL showed optimum pH in an acidic range of 5-5.5. It was highly sensitive to metal-chelating agents such as EDTA, and the lost MGL activity was restored to the original level by the addition of divalent cations such as Ca2+ or Mg2+. The purified MGL was strictly dependent on Ca2+ and underwent rapid denaturing precipitation on Ca2+ depletion even in the presence of detergent. This communication can be the first one dealing with the solubilization, purification and properties of MGL. PMID- 19129637 TI - Structural analysis of a bioactive polysaccharide, PISP1, from the medicinal mushroom Phellinus igniarius. AB - PISP1, a heteropolysaccharide isolated from fruiting bodies of Phellinus igniarius by hot aqueous extraction and purified by DEAE-Sepharose anion-exchange and gel filtration chromatography, is composed of fucose, galactose, mannose, and 3-O-Me-galactose in a ratio of 1:2:1:2. Methylation, monosaccharide analysis, and NMR studies (1H NMR, 13C NMR, COSY-45 degrees , TOCSY, ROESY, HSQC, and HMBC) revealed that PISP1 had a backbone consisting of 1,6-disubstituted-3-O-Me-alpha-D galactopyranosyl residue, 1,3,6-trisubstituted-alpha-D-manopyranosyl residue, 1,4 disubstituted-alpha-D-galactopyranosyl residue, and 1,2-disubstituted-alpha-D galactopyranosyl residue, and had a 1-substituted-alpha-L-fucopyranosyl terminal attached to O-3 of a manopyranosyl residue. Preliminary bioactivity tests conducted in vitro revealed that PISP1 stimulated the proliferation of mouse spleen lymphocytes. PMID- 19129638 TI - Hypocholesterolemic effect of peanut skin and its fractions: a case record of rats fed on a high-cholesterol diet. AB - Peanut skin (PS) is characterized by almost exclusively consisting of polyphenols and fiber. We fractionated PS into a water-soluble fraction (WSF) and water insoluble fraction (WIF), and further fractionated WSF into a soluble dietary fiber fraction (DF) and dietary fiber-free, water-soluble fraction (DFF-WSF). Male Sprague-Dawley rats were fed on high-cholesterol diets supplemented with PS and its fractions. PS, WSF, and DFF-WSF decreased the serum lipid and cholesterol levels and increased those in feces. This effect was probably due to the polyphenols that inhibited intestinal cholesterol absorption. PMID- 19129639 TI - Antimicrobial activity of stereoisomers of morinols a and B, tetrahydropyran sesquineolignans. AB - The antimicrobial activity of all stereoisomers of morinols A and B was tested. All stereoisomers of morinols A and B showed antifungal activity against Alternaria alternata, especially (-)-morinol B which showed the strongest activity. The natural component, (+)-morinol A, and unnatural stereoisomer, (7S,7'S,8R,8'R)-morinol B, showed antibacterial activity against the gram positive bacteria, Bacillus subtilis and Listeria denitrificans. PMID- 19129640 TI - Isolation and absolute stereochemistry of optically active sydonic acid from Glonium sp. (Hysteriales, Ascomycota). AB - Optically active sydonic acid (1) was isolated for the first time from a culture broth of Glonium sp. The absolute stereochemistry was established to be (S) by comparing the circular dichroism (CD) spectrum with that of (+)-curcutetraol after conversion into (+)-sydonol (2). PMID- 19129641 TI - Discovery of novel 2',3',4'-trihydroxy-2-phenylacetophenone derivatives as anti gram-positive antibacterial agents. AB - A number of 2',3',4'-trihydroxy-2-phenylacetophenone derivatives were synthesized and examined for growth inhibition of several kinds of bacteria. 2',3',4' Trihydroxy-2-phenylacetophenone itself exhibited no antibacterial activity, but some of its derivatives showed various antibacterial activities depending on functional groups introduced on the 2-phenyl ring. Eighteen out of 24 compounds synthesized in this study appeared to possess antibacterial activities against at least two Gram-positive strains of Bacillus subtilis and Staphylococcus aureus, 2 (biphenyl-4-yl)-2',3',4'-trihydroxyacetophenone being the most active with LC(50) of 5.8 muM and 5.6 muM respectively. However, none of the synthesized compounds exhibited inhibitory effects on Gram-negative strains, such as Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, and Salmonella enterica, suggesting that anti-Gram-positive specificity of the antibacterial compounds. PMID- 19129642 TI - Long-term survival of Escherichia coli lacking the HipBA toxin-antitoxin system during prolonged cultivation. AB - We designed and constructed six major toxin-antitoxin disruptants (DeltachpBIK, DeltadinJ-yafQ, DeltahipBA, DeltamazEF, DeltarelBE, and DeltayefM-yoeB) of Escherichia coli K-12 W3110. On prolonged cultivation of these disruptants with minimal M9 medium, the DeltahipBA cells exhibited a significantly longer life span than that of the other disruptants and of wild-type cells, as analyzed with a LIVE/DEAD BacLight kit (Invitrogen, Carlsbad, CA) in combination with flow cytometry analysis. The gene expression level of hipA in the wild-type cells was highest at the stationary phase of 40 h. The DeltahipBA cells showed higher macromolecular synthesis activity than the wild-type cells at the stationary phase. Stationary phase cells of DeltahipBA and the wild-type strain showed a significantly extended life span under anaerobic conditions. Furthermore, the DeltahipBA cells showed higher resistance to H(2)O(2) than the wild type. These results suggest that HipBA induces cell death with oxidative stress during prolonged cultivation. This is the first report that an E. coli toxin-antitoxin (TA) system affects frequency of survival during the long-term stationary phase. PMID- 19129643 TI - Identification and characterization of two alternative sigma factors of RNA polymerase in the deep-sea piezophilic bacterium Shewanella violacea, strain DSS12. AB - Two genes for alternative sigma factors, sigma(E2) and sigma(E3), classified in the extracytoplasmic function sigma family for RNA polymerases, were identified in the deep-sea piezophilic bacterium Shewanella violacea DSS12. Amino acid alignments revealed that the domains for transcriptional functions were comparatively conserved compared with Escherichia coli sigma(E) in both proteins. Core-binding analysis suggested that both proteins function as sigma factors. PMID- 19129644 TI - Cloning and characterization of a beta-1,4-mannanase 5C possessing a family 27 carbohydrate-binding module from a marine bacterium, Vibrio sp. strain MA-138. AB - The beta-1,4-mannanase 5C gene (man5C) of Vibrio sp. strain MA-138 was cloned and expressed in Escherichia coli. The man5C gene consisted of 2,010 bp nucleotides encoding a protein of 669 amino acids with a predicted molecular weight of 76,309. beta-1,4-Mannanase (Man5C) is a modular enzyme composed of a catalytic module belonging to glycoside hydrolase family 5, a linker region, and a putative carbohydrate-binding module (CBM) belonging to family 27. Recombinant Man5C exhibited maximal activity at 50 degrees C at pH 7.0, and it had a K(m) of 0.6 mg ml(-1) and a V(max) of 556.2 micromol min(-1) mumol(-1) for glucomannan. Binding studies revealed that the C-terminal putative CBM27 had the ability to bind soluble beta-mannans and contributed to increasing the rate of depolymerization by binding to the polymeric substrate. Man5C of Vibrio sp. MA-138 is the first non-extremophile enzyme to be identified as a beta-mannanase possessing CBM27. PMID- 19129645 TI - Effect of growth temperature and growth phase on the lipid composition of the archaeal membrane from Thermococcus kodakaraensis. AB - Archaea have unique membrane lipids typified by ether linkages of the glycerol-to isoprenoid chains with sn-2,3 stereochemistry that runs against the naturally occurring sn-1,2 stereochemistry of the glycerophospholipids of Bacteria and Eukarya. Membrane lipids were extracted and analyzed from the hyperthermophilic archaeon, Thermococcus kodakaraensis, cultivated at various temperatures. At all growth temperatures examined, both the diphytanylglycerol diether (archaeol, C(20)) and diphytanyldiglycerol tetraether (caldarchaeol, C(40)) were identified as saturated forms, and no other lipids could be identified. The ratio of caldarchaeol to archaeol increased with increasing growth temperature, particularly at 93 degrees C. A larger amount of archaeol was detected from cells in the logarithmic phase than from those in the stationary phase at all temperatures examined. These results indicate that T. kodakaraensis modulated the membrane lipid composition depending on both the growth phase and the growth temperature, and suggest that the membrane fluidity to environmental change was maintained by altering the length of the hydrocarbon chains, and not by side chain saturation such as double-bond hydrogenation nor by such a modification as cyclopentane ring formation. PMID- 19129646 TI - Soluble fibers inhibit carotenoid micellization in vitro and uptake by Caco-2 cells. AB - We evaluated the effects of soluble fibers on beta-carotene and lutein micellization during simulated digestion in vitro, and on carotenoid uptake from mixed micelles by Caco-2 cells. Medium- and high-viscosity alginates and pectins inhibited carotenoid micellization and cellular uptake relative to the fiber-free control. Alginates, carboxy-methylcelluloses, and methylcelluloses inhibited beta carotene uptake mainly by increasing medium viscosity, but pectins might inhibit carotenoid uptake by additional mechanisms. PMID- 19129647 TI - Carbohydrate binding specificity of recombinant human macrophage beta-glucan receptor dectin-1. AB - Human macrophage dectin-1, a type II transmembrane beta-glucan receptor, was expressed as a fusion protein with an N-terminal hexahistidine tag and glutathione S-transferase in an Escherichia coli cell-free translation system, and assayed for binding specificity. Recombinant dectin-1 specifically bound to some beta-glucans, but not to other carbohydrates. The beta-glucan binding of recombinant dectin-1 was inhibited by laminarin, a soluble beta-glucan, and by laminarioligosaccharides, but not by other carbohydrates. These results suggest that recombinant human dectin-1 can be used as a useful probe in identifying ligands in humans and tonic foods due to its strict binding specificity. PMID- 19129648 TI - Analysis of extracellular proteins of Aspergillus oryzae grown on soy sauce koji. AB - Aspergillus oryzae AS 3.951 is widely used in Chinese soy sauce manufacture, but little is known about the profiles of the extracellular proteins from the culture of soybean koji. In this study, we carried out MALDI-TOF/TOF MS analysis of extracellular proteins during koji culture. Besides well-known proteins (TAA and Oryzin), a variety of aminopeptidase and proteases were identical at the proteome level. This suggests that A. oryzae AS 3.951 has a powerful capacity to digest soybean protein. PMID- 19129649 TI - Pyrolysis GC/MS and IR spectroscopy in chitin analysis of molluscan shells. AB - Chitin is an insoluble component in the shells of several molluscan species. It is thought to play important roles, in biomineralization and shell structure. To date, however, reports are scarce and sometimes contradictory, and suffer from methodological problems. Only in a single cephalopod species has the chitin been identified as beta-chitin. We present data on chitin occurrence in 22 species of shell-bearing Mollusca (Conchifera) and Polyplacophora, including the first evidence for scaphopods, based on pyrolysis gas chromatography, mass spectrometry (GC-MS), and infrared spectroscopy (IR). Pyrolysis GC-MS detected chitin in every tested member of the Conchifera. IR spectroscopy before and after chitinase treatment revealed at least three distinct patterns of peak changes. The contents of the insoluble shell organics included not only chitin and proteins, but also insoluble polysaccharides, e.g., glucan. We conclude that chitin was present in the last common ancestor of the Conchifera and that its abundance in the shell matrix depends on the differentiation of the shell. PMID- 19129650 TI - Response to hypoxia, reduction of electron acceptors, and subsequent survival by filamentous fungi. AB - Filamentous fungi usually inhabit normoxic environments by utilizing oxygen as a substrate for respiration and for the biosynthesis of some essential cellular components. This review examines the metabolic mechanisms used by filamentous fungi under oxygen-limited (hypoxic) conditions. Denitrification is one mechanism through which Fusarium oxysporum and other fungi reduce nitrate or nitrite to nitrous oxide, generating nitric oxide as a reaction intermediate. The involvement of cytochrome P450nor as a nitric oxide reductase is a unique feature of fungal denitrification, as opposed to cytochrome bc-type nitric oxide reductase, which is unique to the bacterial mechanism. Ammonia fermentation is the mechanism through which nitrate is reduced to ammonium, and it allows fungal growth under hypoxic conditions. Studies of the model filamentous fungus Aspergillus nidulans have revealed that niaD and niiA encoding NAD(P)H-dependent nitrate and nitrite reductases are essential for producing ammonia. Since niaD and niiA have been identified as genes for nitrate utilization by the fungus, ammonia fermentation and nitrate utilization mechanisms probably share a nitrate reducing mechanism. I also discuss recent progress in studies of the hypoxic response of A. nidulans. PMID- 19129651 TI - Isolation and molecular cloning of a major wheat allergen, Tri a Bd 27K. AB - Tri a Bd 27K is the predominant allergen in wheat. In the present study, this allergen was purified to homogeneity from wheat flour. The N-terminal amino acid sequences of the purified allergen and the peptides obtained by its digestion, with trypsin were determined, and the allergen was shown to be a glycoprotein with an Asn-linked sugar moiety containing fucose residues. A cDNA encoding the allergen was obtained by polymerase chain reaction (PCR). The cDNA codes for a protein of 203 amino acid residues, with a molecular mass of 22,803 Da, that has two tentative sites glycosylated at Asn residues. Homology analysis suggested that the allergen might belong to a family of gamma-interferon-inducible thiol reductases. The cDNA was expressed as a fusion protein with glutathione S transferase in Escherichia coli. However, unlike the allergen purified from wheat, recombinant Tri a Bd 27K was not immunoblotted with IgE antibodies in the serum of a wheat-sensitive patient. PMID- 19129652 TI - Induction of differentiation in k562 cell line by nicotinic acid-related compounds. AB - Nicotinic acid and nicotinamide belong to the water-soluble vitamins, and they have many physiological and pharmacological functions in various organisms. In this study, we investigated the differentiation-inducing ability of nicotinic acid-related compounds in chronic myelogenous leukemia K562 cell line. Proliferation of K562 leukemia cells was inhibited by several nicotinic acid related compounds. Hemoglobin content was increased by nicotinic acid and by isonicotinic acid. Isonicotinic acid increased gamma-globin mRNA expression as much as sodium butyrate did. The nuclei of nicotinic acid and of isonicotinic acid-treated cells decreased in size and the chromatin became more condensed. It was verified that nicotinic acid and isonicotinic acid induced erythroid differentiation in K562 cells. Expression of glycophorin A was increased by sodium butyrate. In contrast, it was decreased by nicotinic acid and by isonicotinic acid, suggesting that these compounds differentiate K562 to erythrocytes through different pathways than sodium butyrate does. Our data perhaps provide useful information as to the mechanisms of cell differentiation. PMID- 19129654 TI - Cloning and transcript analysis of multiple genes encoding the glycoside hydrolase family 6 enzyme from Coprinopsis cinerea. AB - We searched the genome database of the basidiomycete Coprinopsis cinerea (Coprinus cinereus) and found five genes encoding the glycoside hydrolase family 6 (GH6) enzyme, CcCel6A, CcCel6B, CcCel6C, CcCel6D, and CcCel6E, designated in order of increasing locus number (CC1G_01107.1, CC1G_04166.1, CC1G_08276.1, CC1G_08277.1, and CC1G_10605.1). The amino acid sequence of CcCel6A suggests a two-domain structure consisting of an N-terminal family 1 carbohydrate-binding module (CBM1) and a GH6 catalytic domain, while the other genes lack CBM1. The transcripts of CcCel6A were observed at the active growth stage in cellulose culture, whereas they were absent from glucose culture. Cellobiose strongly induced transcription of CcCel6A. On the other hand, transcripts of CcCel6B, -D, and -E were detected in both glucose and cellulose cultures, and transcription of them was induced weakly by cellobiose. The transcript level of CcCel6C was not influenced by glucose or cellobiose. PMID- 19129653 TI - Phosphocholine-containing glycosyl inositol-phosphoceramides from Trichoderma viride induce defense responses in cultured rice cells. AB - We isolated two major zwitterionic glycosphingolipids (ZGLs) from the phytopathogenic filamentous fungus Trichoderma viride. Structural analyses showed that the ZGLs (designated Tv-ZGL2 and Tv-ZGL3) were the same as the glycosphingolipids ZGL2 and ZGL4 from Acremonium sp., which are described in our previous paper. ZGLs have the following structure: Man(alpha1-6)GlcN(alpha1-2)Ins P-Cer (Tv-ZGL2) and phosphocholine (PC)-->6Man(alpha1-6)GlcN(alpha1-2)Ins-P-Cer (Tv-ZGL3). To determine whether these ZGLs have functional roles in plant-fungus interaction, we tested to determine whether they would induce defense responses in cultured rice cells. We found that T. viride's ZGLs elicited expression of the PAL and PBZ1 genes, both of which are associated with pathogen resistance. Tv ZGL2 induced cell death at a moderate rate. Tv-ZGL3, which contains a PC moiety, induced a high level of cell death in rice cells. PMID- 19129655 TI - Stabilization of glycoprotein liquid formulation using arginine: a study with lactoferrin as a model protein. AB - The formulation of new biotherapeutics without human serum albumin (HSA) could decrease the potential risk of blood-transmitted diseases and those caused by infectious viruses and other pathogens. In the present study, arginine was examined as a potential alternative to HAS, and bovine lactoferrin (bLf) was used as a representative model glycoprotein since bLf has potential immunomodulatory and antiviral activity. The optimal formulation for the mixture was determined to be 10 mM arginine, 15% (w/v) trehalose, and 0.02% (v/v) Tween 80, using a statistical analysis program, Minitab. Analyses were performed using reverse phase high-performance liquid chromatography (HPLC) and SDS-PAGE. The blf HSA free formulations lost only 12-20% of blf compared with 46% for control (without additives) after 28 d of storage. Based on long-term stability studies, the HSA free formulation developed in this study had a stronger effect on the stability of bLf (1.4-fold) than HSA formulation under various storage conditions over 6 months. PMID- 19129656 TI - Crystallization and structural analysis of cytochrome c(6) from the diatom Phaeodactylum tricornutum at 1.5 A resolution. AB - We determined for the first time the crystal structure of diatom cytochrome c(6) from Phaeodactylum tricornutum at 1.5 A resolution. The overall structure of the protein was classified as a class I c-type cytochrome. The physicochemical properties of the protein were examined by denaturation with guanidine hydrochloride and urea, and compared with those of other algal cytochrome c(6). PMID- 19129657 TI - beta-glucosidase catalyzed syntheses of pyridoxine glycosides. AB - Enzymatic syntheses of pyridoxine glycosides were carried out in di-isopropyl ether organic medium using beta-glucosidase isolated from sweet almond. Optimum conditions determined for the reaction with D-glucose were 40% (w/w D-glucose) beta-glucosidase at 0.18 mM (1.8 ml) of pH 5 acetate buffer over a 72 h incubation period. Of 11 carbohydrates employed, beta-glucosidase gave 7-O-(alpha D-glucopyranosyl)pyridoxine 5a, 7-O-(beta-D-glucopyranosyl)pyridoxine 5b, 6-O (alpha-D-glucopyranosyl)pyridoxine 5c, 7-O-(alpha-D-galactopyranosyl)pyridoxine 6a, 7-O-(beta-D-galactopyranosyl)pyridoxine 6b, 6-O-(alpha-D galactopyranosyl)pyridoxine 6c, 7-O-(alpha-D-mannopyranosyl)pyridoxine 7a, 7-O (beta-D-mannopyranosyl)pyridoxine 7b, and 6-O-(alpha-D-mannopyranosyl)pyridoxine 7c in yields ranging from 23 to 40%. PMID- 19129658 TI - Vacuolar sorting behaviors of 11S globulins in plant cells. AB - Plant seed cells amass storage proteins that are synthesized on the endoplasmic reticulumn (ER) and then transported to protein storage vacuoles (PSVs). Many dicotyledonous seeds contain 11S globulin (11S) as a major storage protein. We investigated the accumulation behaviors of pea and pumpkin 11S during seed maturation and compared them with soybean 11S biogenesis (Mori et al., 2004). The accumulation of pea 11S in seeds was very similar to that of soybean 11S at all the development stages we examined, whereas pumpkin 11S condensed in the ER. The determinant of accumulation behavior might be the surface hydrophobicity of 11S. Further, we examined the accumulation of 11Ss in tobacco BY-2 cells to analyze behavior in the same environment. 11Ss expressed in BY2 cells were all observed in precursor form (pro11S). Pro11S with high surface hydrophobicity might be transported to vacuoles in a multivesicular body-mediated pathway when the expression level remains low. PMID- 19129659 TI - The Arabidopsis G-protein beta-subunit is required for defense response against Agrobacterium tumefaciens. AB - Typical early pathogen-associated molecular pattern (PAMP) responses include the generation of reactive oxygen species (ROS) and MAP kinase (MAPK) activation, but little is known about the molecular mechanisms that link receptor activation to intracellular signal transduction. In this study, we found that in agb1-2 (AGB1 null mutation) mutants, ROS production triggered by flg22 or elf18 was significantly reduced and that elf18-stimulated PAMP-triggered immunity (PTI) against Agrobacterium tumefaciens was impaired. Thus AGB1 appears to integrate PAMP perception into downstream ROS production, and also to transmit the EF-Tu signal to the defense response, leading to reduced transformation by A. tumefaciens. PMID- 19129660 TI - Prenyl alcohol production by expression of exogenous isopentenyl diphosphate isomerase and farnesyl diphosphate synthase genes in Escherichia coli. AB - Isopentenyl diphosphate isomerase (idi) and farnesyl diphosphate synthase (ispA) genes were overexpressed in Escherichia coli. The resulting transformant showed 6.8-fold higher production of farnesol (389 microg/l). In a similar manner, overexpression of idi and mutated ispA led to high production of geranylgeraniol (128 microg/l). PMID- 19129661 TI - Induction of flowering by inducers of systemic acquired resistance in the Lemna plant. AB - In plants, salicylic acid induces both systemic acquired resistance (SAR) and flowering. We found that flower induction in the Lemna species was promoted by reactive oxygen species-generating conditions and SAR inducers. The present paper suggests that SAR in plants and flowering in Lemna share common inducers. PMID- 19129662 TI - Alternative processing of proproteins in Aspergilli kexB gene disruptants under hyperosmotic conditions. AB - Disruption of the kexB gene encoding a subtilisin-like processing protease in Aspergillus oryzae and Aspergillus nidulans led to remarkable morphological defects, and these phenotypes were suppressed under hyperosmotic conditions. In this study, we investigated to determine whether non-KexB proteases might complement the in vivo function of KexB in the two Aspergillus kexB disruptants. Neither overexpression of opsA or opsB encoding A. oryzae aspartyl proteases homologous to yeast yapsins (YPS1/2) suppressed the kexB mutation, although yapsins are multicopy suppressors for the yeast kex2 mutation. A. nidulans and A. oryzae kexB disruptants grown under hyperosmotic conditions processed a recombinant fusion protein carrying a synthetic dibasic processing site (Lys-Arg) although the disruptants grown under normal growth conditions did not cleave the site. These results suggest that the two Aspergilli have other potential processing proteases that are induced and/or activated under hyperosmotic conditions and consequently complement, at least in part, the in vivo function of KexB. PMID- 19129663 TI - Extracellular enzymes produced by marine eukaryotes, thraustochytrids. AB - Extracellular enzymes produced by six strains of thraustochytrids, Thraustochytrium, Schizochytrium, and Aurantiochytrium, were investigated. These strains produced 5 to 8 kinds of the extracellular enzymes, depending on the species. Only the genus Thraustochytrium produced amylase. When insoluble cellulose was used as substrate, cellulase was not detected in the six strains of thraustochytrids. This study indicates that marine eukaryotes, thraustochytrids, produced a wide variety of extracellular enzymes. PMID- 19129664 TI - The Effect of Secoisolariciresinol on 3T3-L1 Adipocytes and the Relationship between Molecular Structure and Activity. AB - As we have reported, flaxseed lignan, (+)-secoisolariciresinol (SECO), (-)-SECO, and meso-SECO were stereoselectively synthesized and their biological functions were evaluated. In the present study, we focused on the effects of SECOs on the regulation of 3T3-L1 adipocytes, and identified the structure-activity relationships. Optically active SECO and meso-SECO were tested for their effects on lipid metabolism in 3T3-L1 adipocytes. (-)-SECO accelerated adiponectin production of 3T3-L1 adipocytes. On the other hand, (+)- and meso-SECO suppressed the production of adiponectin. In addition, triglyceride (TG) accumulation in 3T3 L1 adipocytes was significantly suppressed by all three SECOs tested here, as was 17beta-estradiol, when the SECOs were added to the medium during induction of 3T3 L1 preadipocytes to adipocytes. Especially, (-)-SECO strongly reduced TG accumulation. It is well-known that SECO has estrogen-like activity. Hence the estrogen-like activity of each SECO compound was assessed. Only (-)-SECO had estrogen-like activity. PMID- 19129665 TI - Gene expression profile in the anterior regeneration of the earthworm using expressed sequence tags. AB - In order to gain insight into the gene expression profiles associated with anterior regeneration of the earthworm, Perionyx excavatus, we analyzed 1,159 expressed sequence tags (ESTs) derived from cDNA library early anterior regenerated tissue. Among the 1,159 ESTs analyzed, 622 (53.7%) ESTs showed significant similarity to known genes and represented 338 genes, of which 233 ESTs were singletons and 105 ESTs manifested as two or more ESTs. While 663 ESTs (57.2%) were sequenced only once, 308 ESTs (26.6%) appeared 2 to 5 times, and 188 ESTs (16.2%) were sequenced more than 5 times. A total of 803 genes were categorized into 15 groups according to their biological functions. Among 1,159 ESTs sequenced, we found several gene encoding signaling molecules, such as Notch and Distal-less. The ESTs used in this study should provide a resource for future research in earthworm regeneration. PMID- 19129666 TI - Changes in thiamin contents in porcine muscles and liver during growth. AB - Changes in the thiamin contents in three types of porcine muscle and porcine liver during growth were investigated. The muscular thiamin content was lower at the newborn stage than at fetal stage, and increased after the weaning period. The liver thiamin content, however, remained unchanged from the fetal stage to 5 months old. The changes in thiamin contents were different between Landrace and Meishan pigs. PMID- 19129667 TI - Purification and characterization of a major collagenase from Streptomyces parvulus. AB - A major collagenase was purified about 96-fold from a crude enzyme sample of Streptomyces parvulus by chromatography on Q-Sepharose, Sephacryl S-200, and butyl-Toyopearl. The purified enzyme showed a relative molecular mass of approximately 52,000 on SDS-PAGE and a pH optimum at about 9.0, and was strongly inhibited by metal-chelating agents. It also cleaved 4-phenylazobenzyloxycarbonyl Pro-Leu-Gly-Pro-D-Arg specifically at the Leu-Gly bond, with a K(m) value of 0.60 mM at pH 9.0 at 37 degrees C. Based on the amino acid sequences of the N-terminal region and internal tryptic peptides, the corresponding gene was cloned. The DNA sequence of the cloned gene indicated that the enzyme is produced as an 864 residue precursor protein with a 408-residue prepro sequence followed by a 456 residue mature enzyme moiety. The enzyme is most homologous with the collagenase from S. coelicolor, the identity being 73%, and it is thought to be a member of the Vibrio collagenase subfamily. PMID- 19129668 TI - Identification of Japanese and chinese green tea cultivars by using simple sequence repeat markers to encourage proper labeling. AB - To identify commercial Japanese monovarietal green tea and imported green tea samples, leading Japanese cultivars were fingerprinted by using six simple sequence repeat markers analyzed by a capillary sequencer. Two well-authenticated imported Chinese monovarietal green tea samples were also fingerprinted by the same markers, one of which, was Fuyun, was a clonally propagated cultivar, and the other, Jiukengzhong, was seed-propagated. At least three markers used in this study identified 16 leading Japanese cultivars and Fuyun. Although Jiukengzhong was a mixed population with diverse genotypes, some individuals had a unique allele in one simple sequence repeat marker that was not detected in the 16 leading Japanese cultivars, an additional 39 cultivars, and Fuyun. This allele was effective as a detection marker for Jiukengzhong. These results support the use of simple sequence repeat markers for the identification of Japanese monovarietal green tea and also of imported green tea made from foreign cultivars. PMID- 19129669 TI - Efficiency of sodium hypochlorite and calcinated calcium in killing Escherichia coli O157:H7, Salmonella spp., and Staphylococcus aureus attached to freshly shredded cabbage. AB - The effects of the disinfectants NaClO and calcinated calcium on the food-borne pathogens Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella spp. attached to shredded cabbage leaves were examined. After these bacteria were attached to shredded leaves for 1 h, the leaves were treated with NaClO and/or calcinated calcium. About 2.6-log and 3.5-log reductions of E. coli O157 were achieved by treatment with NaClO (100 ppm, pH 6.0, 10 min) and calcinated calcium (0.1%, 20 min), respectively. The combination of 100 ppm NaClO and 0.1% calcinated calcium resulted in a 3- to 4-log reduction in the pathogen populations without apparent deteriorative effects. The bacterial numbers in the treated cabbage did not increase during storage at 4 degrees C. However, sensory evaluation including appearance and flavor indicated that the quality of the treated cabbage declined during storage. In conclusion, the combination of NaClO and calcinated calcium was useful in treatment before eating. PMID- 19129670 TI - Antitumor effects of a combination of interferon-alpha and sorafenib on human renal carcinoma cell lines. AB - To support the role of interferon (IFN)-alpha and sorafenib combination therapy against renal cell carcinoma (RCC), the effects of IFN-alpha and sorafenib on tumor growth, vascular endothelial growth factor (VEGF) production, and phosphorylation levels of extracellular signal-regulated kinase (ERK) and mitogen activated protein/ERK kinase (MEK) were examined using several cultured RCC cell lines (ACHN, Caki-1, Caki-2, SMKT-R1, SMKT-R2, SMKT-R3 and SMKT-R4). IFN-alpha or sorafenib alone inhibited the proliferation of all the cell lines except Caki-2, while combined treatment with the two agents showed enhanced inhibitory effects compared to treatment with each agent alone. VEGF production was inhibited by IFN alpha alone in ACHN and SMKT-R2 cells and by sorafenib alone in ACHN, Caki-1, SMKT-R1 and SMKT-R2 cells. However, sorafenib increased VEGF production by Caki-2 cells. Interestingly, combined treatment with the two agents suppressed VEGF production by SMKT-R1 and SMKT-R2 cells more strongly than IFN-alpha or sorafenib alone. Although phosphorylated ERK (p-ERK) was increased after 30 min of treatment with IFN-alpha alone, no difference was observed between control and IFN-alpha-treated cells after 2 h. Sorafenib decreased p-ERK in ACHN, Caki-1, SMKT-R1 and SMKT-R2 cells, but increased p-ERK in Caki-2, SMKT-R3 and SMKT-R4 cells, after 2 h. Combined treatment with IFN-alpha and sorafenib decreased p-ERK compared to treatment with each agent alone in all cell lines except Caki-2. However, IFN-alpha did not inhibit the p-ERK increase induced by sorafenib in Caki-2 cells. Phosphorylated MEK showed similar patterns to p-ERK after the various treatments. In conclusion, combined treatment with IFN-alpha and sorafenib suppressed cell proliferation and VEGF production more strongly than treatment with each agent alone in several RCC cell lines. PMID- 19129671 TI - The possible involvement of tensin2 in the expression and extension of nephrin by glomerular podocytes in mice. AB - The prevalence of chronic kidney disease (CKD) is increasing worldwide and proteinuria is a critical prognostic indicator of CKD. Nephrin is produced by podocytes and functions as a slit barrier for inhibition of proteinuria. Nephrin expression is frequently decreased in CKD patients. Nevertheless, the mechanism by which nephrin declines during CKD-related pathological states remains to be determined. Using tensin2-deficient mice (ICGN/Oa strain), we provide evidence that tensin2 is important for glomerular nephrin expression in vivo. In heterozygous mice with a single mutated tensin2 allele, nephrin expression was maintained, while albuminuria was not observed. In contrast, nephrin expression was impaired, especially in the central zones of glomeruli of homozygous mice (with double mutated tensin2 alleles), even at one week after birth. In homozygous mice, extension of synaptopodin, a key actin-associated protein, was also suppressed in the central zone of glomerular tufts. Consistent with the loss of nephrin and synaptopodin expression, severe albuminuria was detected in homozygous ICGN/Oa mice. Therefore, we suggested that tensin2 is involved in expression and extension of nephrin, while tensin2 deficiency may result in proteinuria, associated with the loss of slit integrity. PMID- 19129672 TI - Topographic study on nerve-associated lymphatic vessels in the murine craniofacial region by immunohistochemistry and electron microscopy. AB - The distribution and fine structure of lymphatic vessels associated with nerves was studied by immunohistochemistry in the murine craniofacial region. The tissue sections and blocks were immunostained for LYVE-1, protein gene product 9.5, CD34 and aquaporin-1 to demonstrate the lymphatic vessels, nerves, blood vessels and water channel protein, respectively. Transmission electron microscopic examination was also performed to investigate the relationship between the lymphatics and nerves. In the nasal area, the lymphatics were found in dura mater on the cribriform plate and beneath the nasal mucosa, this supposedly supplying the cerebrospinal fluid drainage route along the olfactory nerves. The proximal portions of the cranial nerves were equipped with the lymphatics in the epineurium. In the distal portions of the nerves, the lymphatics were distributed in close proximity of the perineural sheath, and thus might contribute to maintenance of microenvironment suitable for the nerves by an absorptive activity of the lymphatic endothelial cells. The present findings suggest that the lymphatic system associated with the cranial nerves provides the pathway for transport of cerebrospinal fluid, tissue fluid, and free cells involved in immune response and tumor metastasis in the craniofacial region. PMID- 19129673 TI - Histochemical demonstration of a monocarboxylate transporter in the mouse perineurium with special reference to GLUT1. AB - Peripheral nerves express GLUT1 in both endoneurial blood vessels and the perineurium and utilize glucose as a major energy substrate, as does the brain. However, under conditions of a reduced utilization of glucose, the brain is dependent upon monocarboxylates such as ketone bodies and lactate, being accompanied by an elevated expression of a monocarboxylate transporter (MCT1) in the blood-brain barrier. The present immunohistochemical study aimed to examine the expression of MCT1 in the peripheral nerves of mice. MCT1 immunoreactivity was found in the perineurial sheath and colocalized with GLUT1, while the endoneurial blood vessels expressed GLUT1 only. An intense expression of MCT1 in the perineurium was confirmed by Western blot and in situ hybridization analyses. Ultrastructurally, the MCT1 and GLUT1 immunoreactivities in the thick perineurium showed an intensity gradient decreasing towards the innermost layer. In neonates, the MCT1 immunoreactivity in the perineurium was intense, while the GLUT1 immunoreactivity was faint or absent. These findings suggest that peripheral nerves depend on monocarboxylates as a major energy source and that MCT1 in the perineurium is responsible for the supply of monocarboxylates to nerve fibers and Schwann cells. PMID- 19129674 TI - Regulation of volume-regulated outwardly rectifying anion channels by phosphatidylinositol 3,4,5-trisphosphate in mouse ventricular cells. AB - Volume-regulated outwardly rectifying anion channel (VRAC) plays an important role in cell-volume regulation in many types of cells. Little is known about the regulation of VRAC by phosphatidylinositides (PIs), which include phosphatidylinositol 3,4,5-trisphosphate (PIP3) and phosphatidylinositol 4,5 bisphosphate (PIP2). We examined the effect of PIs on the VRAC current activated in hypotonic solution in mouse ventricular cells. VRAC current was inhibited strongly by intracellular application of LY294002 (a phosphatidylinositol 3 kinase (PI3K) inhibitor) or anti-PIP3 antibody (PIP3-Ab), and less strongly by anti-PIP2 antibody (PIP2-Ab). LY294002 inhibited regulatory volume decrease in hypotonically swollen cells, which was in parallel with the VRAC inhibition. Intracellular PIP3 or PIP2 influenced neither the basal background current in isotonic solution nor the VRAC current in hypotonic solution. However, PIP3, but not PIP2, restored the VRAC current suppressed by LY294002 or PIP2-Ab. These results suggest that the activation of VRAC current requires the presence of intracellular PIP3, that PI3K-mediated increase in PIP3 level is sufficient to fully activate VRAC current, and that PIP3 alone without osmotic stimulation cannot induce VRAC current. We propose that VRAC in mouse ventricular cells is regulated by PIP3 and/or its down stream signaling pathways. PMID- 19129675 TI - The facial massage reduced anxiety and negative mood status, and increased sympathetic nervous activity. AB - The aim of this study was to clarify the effects of 45 min of facial massage on the activity of autonomic nervous system, anxiety and mood in 32 healthy women. Autonomic nervous activity was assessed by heart rate variability (HRV) with spectral analysis. In the spectral analysis of HRV, we evaluated the high frequency components (HF) and the low- to high-frequency ratio (LF/HF ratio), reflecting parasympathetic nervous activity and sympathetic nervous activity, respectively. The State Trait Anxiety Inventory (STAI) and the Profile of Mood Status (POMS) were administered to evaluate psychological status. The score of STAI and negative scale of POMS were significantly reduced following the massage, and only the LF/HF ratio was significantly enhanced after the massage. It was concluded that the facial massage might refresh the subjects by reducing their psychological distress and activating the sympathetic nervous system. PMID- 19129676 TI - Suppressive action of pituitary adenylate cyclase activating polypeptide (PACAP) on proliferation of immature mouse Leydig cell line TM3 cells. AB - We aimed in the current study to understand the participation of PACAP in stage specific Leydig and Sertoli cell functions. For this purpose, clonal cell lines TM3 (Leydig) and TM4 (Sertoli) cells, derived from the testis of immature BALB/c mice, were used. PACAP-specific receptors were detected in TM3 cells, but not in TM4 cells, which were characterized as PAC1 (type I) receptors. Stimulation of cAMP accumulation and testosterone secretion were observed in TM3 cells during 1 2 h treatment with PACAP38 (10(-10)-10(-7) M) or PACAP27 (10(-11)-10(-7) M). After around 10 h treatment with 10(-11)-10(-7) M PACAP38 or PACAP27, proliferation of TM3 cells was suppressed in time- and dose-dependent manners, which was confirmed by real-time cell electronic sensing (RT-CES) system and phase-contrast microscopy. At 6 h after the addition of PACAP38, the percent cell population in G(2)/M phases increased significantly, while that in S phase showed significant decrease with little change in G(0)/G(1) phases. The results revealed that PACAP exerts, in addition to early stimulatory effect on cAMP formation steroidogenesis, sustained suppressive effect on cell proliferation in TM3 cells by controlling progression of the cell cycle. The suppressive action of PACAP on proliferation in TM3 cells supports the stage-specific participation of the peptide in differentiation of immature mouse Leydig cells. PMID- 19129677 TI - Fucoidan inhibits parainfluenza virus type 2 infection to LLCMK2 cells. AB - The effects of fucoidan and L-fucose, a fundamental major component of fucoidan, on the growth of human parainfluenza virus type 2 (hPIV-2) in LLCMK(2) cells were investigated. Fucoidan inhibited cell fusion and hemadsorption, but L-fucose only partly inhibited both. Virus RNA was not detected in the hPIV-2 infected cells cultured with fucoidan. However, L-fucose did not inhibit virus RNA synthesis. Indirect immunofluorescence study showed that virus protein synthesis was inhibited by fucoidan, but not by L-fucose. Furthermore, using a recombinant, green fluorescence protein-expressing hPIV-2, it was found that virus entry was inhibited by fucoidan, but not by L-fucose. These results suggested that fucoidan inhibited virus adsorption to the surface of the cells by binding to the cell surface and prevented infection, indicating that the sulfated polysaccharide form was important for the inhibition by fucoidan. PMID- 19129678 TI - Complications in the catheter ablation of atrial fibrillation: incidence and management. AB - Radiofrequency catheter ablation is widely performed as an effective treatment for recurrent, drug-resistant atrial fibrillation (AF). Recently, various types of ablation strategies, such as extensive encircling pulmonary vein (PV) isolation, linear ablation and complex fractionated electrogram-guided ablation, have been used to achieve a high efficacy for all types of AF. However, several complications (thromboembolic events, PV stenosis and atrio-esophageal fistula etc) with an incidence of 3.9-6%, have been reported. It is important to understand the incidence, prevention and management of complications in order to develop safe ablation strategies. PMID- 19129679 TI - Peroxisome proliferator-activated receptor gamma and cardiovascular diseases. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptor superfamily and form heterodimers with retinoid X receptor. Three PPAR isoforms have been isolated and termed alpha, beta (or delta) and gamma. Although PPARgamma is expressed predominantly in adipose tissue and associated with adipocyte differentiation and glucose homeostasis, PPARgamma is also present in a variety of cell types. Synthetic antidiabetic thiazolidinediones (TZDs) are well known as ligands and activators for PPARgamma. After it was reported that activation of PPARgamma suppressed production of pro-inflammatory cytokines in activated macrophages, medical interest in PPARgamma has grown and there has been a huge research effort. PPARgamma is currently known to be implicated in various human chronic diseases such as diabetes mellitus, atherosclerosis, rheumatoid arthritis, inflammatory bowel disease, and Alzheimer's disease. Many studies suggest that TZDs not only ameliorate insulin sensitivity, but also have pleiotropic effects on many tissues and cell types. Although activation of PPARgamma seems to have beneficial effects on cardiovascular diseases, the mechanisms by which PPARgamma ligands prevent their development are not fully understood. Recent data about the actions and its mechanisms of PPARgamma dependent pathway in cardiovascular diseases are discussed here. PMID- 19129680 TI - A receptor-independent effect of estrone sulfate on the HERG channel. AB - We recently reported that physiological concentrations of 17beta-estradiol partially down-regulate cardiac rapidly-activating delayed rectifier K(+) currents (hERG currents) independently of estrogen-receptor signaling. To determine if other estrogens have the same effect as that of 17beta-estradiol, we investigated receptor-independent effects of estrone, estrone 3-sulfate, and estriol on hERG currents in patch-clamped estrogen-negative HEK293 cells. Only estrone 3-sulfate partially suppressed hERG currents in a receptor-independent manner by modifying the gating. The concentration-dependence of estrone 3-sulfate revealed that physiological levels of circulating estrone 3-sulfate can modulate hERG currents to the maximal extent in both women and men at any age. PMID- 19129681 TI - Nicotine-induced neurogenic relaxation in the mouse colon: changes with dextran sodium sulfate-induced colitis. AB - Nicotine has been shown to reduce both tone and muscular activity in the human colon by releasing nitric oxide (NO) from nerves. To our knowledge, however, the effect of nicotine on mouse colon has not been elucidated, and the response in tissue from ulcerative colitis (UC) has not been investigated. We examined nicotine-induced responses in colon from control mice and mice with dextran sodium sulfate (DSS)-induced UC. In controls, bath application of nicotine caused a transient relaxation in longitudinal preparations from the transverse and distal colons but not from the rectum. The response was observed in the presence of bethanechol, abolished by treatment with tetrodotoxin and hexamethonium, and mediated partially (>50%) by the NO pathway. In longitudinal preparations of the distal colon from DSS-treated mice, spontaneous contractions decreased markedly, and nicotine caused contraction without relaxation in half of the preparations tested. Nicotine-induced relaxation in the presence of bethanechol was significantly decreased in the DSS-treated distal colon without changing bethanechol-induced contractions. These data suggest that 1) responses to nicotine differ dependent on colon regions, 2) DSS treatment predominantly caused nicotine-sensitive neurogenic changes in distal colon, and 3) DSS treatment may reverse the direction of nicotine-evoked responses in the colon, in mice. PMID- 19129682 TI - Possible mechanisms underlying statin-induced skeletal muscle toxicity in L6 fibroblasts and in rats. AB - 3-Hydroxy-3-methylglutaryl CoA reductase inhibitors (statins) are safe and well tolerated therapeutic drugs. However, they occasionally induce myotoxicity such as myopathy and rhabdomyolysis. Here, we investigated the mechanism of statin induced myotoxicity in L6 fibroblasts and in rats in vivo. L6 fibroblasts were differentiated and then treated with pravastatin, simvastatin, or fluvastatin for 72 h. Hydrophobic simvastatin and fluvastatin decreased cell viability in a dose dependent manner via apoptosis characterized by typical nuclear fragmentation and condensation and caspase-3 activation. Both hydrophobic statins transferred RhoA localization from the cell membrane to the cytosol. These changes induced by both hydrophobic statins were completely abolished by the co-application of geranylgeranylpyrophosphate (GGPP). Y27632, a Rho-kinase inhibitor, mimicked the hydrophobic statin-induced apoptosis. Hydrophilic pravastatin did not affect the viability of the cells. Fluvastatin was continuously infused (2.08 mg/kg at an infusion rate of 0.5 mL/h) into the right internal jugular vein of the rats in vivo for 72 h. Fluvastatin infusion significantly elevated the plasma CPK level and transferred RhoA localization in the skeletal muscle from the cell membrane to the cytosol. In conclusion, RhoA dysfunction due to loss of lipid modification with GGPP is involved in the mechanisms of statin-induced skeletal muscle toxicity. PMID- 19129683 TI - Malignancy grade-dependent expression of K+-channel subtypes in human prostate cancer. AB - K(+) channels are key molecules in the progression of several cancer types and considered to be potential targets for cancer therapy. We examined the gene expressions of voltage-gated (K(v)), Ca(2+)-activated (K(Ca)), and two-pore domain (K(2P)) K(+)-channel subtypes in needle-biopsy samples of human prostate cancer (PCa) by real-time PCR and compared them with those in PCa epithelial cell lines. The expression of K(v)1.3, K(Ca)1.1, K(Ca)3.1, and K(2P)1 markedly increased in the PCa group with Gleason score of 5 - 6 (GS5-6) but significantly decreased in the GS8-9 group. This malignancy grade-dependent K(+)-channel expression pattern may provide a convenient marker to understand PCa progression level. PMID- 19129684 TI - Radiographic alveolar bone loss in patients undergoing periodontal maintenance. AB - The aim of the present cross-sectional investigation was to evaluate percentage of bone loss in patients who had been one year under periodontal maintenance at the Department of Periodontology, Faculty of Dentistry, University of Sao Paulo by radiographic analysis. Complete sets of periapical radiographs provided data regarding percentage of alveolar bone loss, which was correlated with arches, tooth group and proximal sites. The sample consisted of 27 men and 53 women ranging in age from 16 to 85 years (mean: 48.3 years). A total of 1,120 periapical radiographs (1,970 teeth) were digitized and analyzed with the Image Tool software (University of Texas Health Science Center). Bone loss was defined as when the distance between the cemento-enamel junction and the alveolar bone crest was greater than 2 mm. Two examiners (p<0.0001) performed radiographic measurements of bone loss. The Greenhouse-Geisser normality test and a univariate analysis of variance were used for statistical analysis. Mean bone loss was 20.60% (+/-12.12). The highest level of bone loss was observed on the distal surface and in the upper arch, as well as in the upper incisors and molars. PMID- 19129685 TI - Inhibitory effect of cranberry polyphenol on cariogenic bacteria. AB - The purpose of this study was to investigate the effect of cranberry polyphenol fraction on mutans streptococci. Hydrophobicity is an important factor in the adherence of bacteria to the tooth surface. We found that cranberry polyphenol fraction significantly decreased the hydrophobicity of Streptococcus sobrinus 6715, Streptococcus mutans MT8148R and JC2 in a dose-dependent manner (p<0.05). Biofilm formation by S. sobrinus 6715 and S. mutans MT8148R was inhibited by 100 microg/ml cranberry polyphenol fraction (p<0.01). When dosage was increased to 500 microg/ml, biofilm formation by S. mutans JC2 was significantly inhibited (p<0.05). Addition of 500 microg/ml cranberry polyphenol fraction to medium inhibited growth of S. mutans MT8148R compared with the control (p<0.05). PMID- 19129686 TI - Static MR images for diagnosis of swallowing. AB - Evaluation of swallowing has been made possible by cine-Magnetic resonance (MR) imaging with high time resolution. However, the spatial resolution in cine-MR imaging remains inadequate for the detection of anatomical structures. Therefore, it is necessary to refer to static MR images in conjunction with cine-MR imaging. The aim of this study was to determine which MR parameters were appropriate for static imaging of the anatomical structures involved in swallowing. MR imaging was carried out, and T1-weighted, T2-weighted and proton-density-weighted MR images were obtained in the sagittal plane in 5 healthy volunteers. Each image was evaluated for anatomic landmark clarity by 3 oral radiologists. The anatomic landmarks selected were the lip, tip of tongue, center of tongue, tongue base, soft palate and epiglottis. Differences in clarity among 3 imaging modalities were evaluated. A 3-point score rating system was used. The results showed that lower TE sequences, i.e., either T1-weighted or proton-density-weighted images, were the most suitable for use in conjunction with cine-MR imaging in diagnosing swallowing disorders. PMID- 19129687 TI - In vitro-evaluation of secondary caries formation around restoration. AB - The objective of this in vitro study was to evaluate demineralization around restorations. Class V preparations were made on the buccal and lingual surfaces of each tooth. TPH (Group 1), Fuji II LC (Group 2), Tetric (Group 3), Dyract (Group 4), GS 80 (Group 5) and Chelon Fil (Group 6) were randomly placed in equal numbers of teeth. The teeth were submitted to a pH-cycling model associated with a thermocycling model. Sections were made and the specimens were examined for the presence of demineralization under polarized light microscopy. Demineralization was significantly reduced with Chelon Fil (Group 6). Furthermore, a similar inhibitory effect on the development of demineralization was observed in Groups 2, 4 and 5. PMID- 19129688 TI - JAK2V617F-positive essential thrombocythemia and multiple myeloma with IGH/CCND1 gene translocation coexist, but originate from separate clones. AB - Overlapping of essential thrombocythemia (ET) and multiple myeloma (MM) has been extremely rare. Our report concerns a case with concomitant ET and MM, where JAK2V617F was present in non-myeloma peripheral blood leukocytes and bone marrow (BM) hematopoietic cells, but not in BM-derived CD138-positive myeloma cells. In contrast, double-color fluorescence in situ hybridization analysis showed that BM derived CD138-positive myeloma cells possessed the gene translocation between the immunoglobulin heavy chain gene and the cyclin D1 gene, which was not involved in non-myeloma hematopoietic cells. This is the first case with concomitant ET and MM in which the 2 hematologic neoplasms were shown to have originated from separate malignant clones at hierarchically different differentiation levels resulting from independent acquisition of different molecular aberrations. Among the 10 reported cases, including ours, ET preceded MM in 8 cases, but MM never preceded ET. We suggest that MM clones may have greater proliferative potency compared with ET clones, and that the treatment modification from ET to MM did not seem to exacerbate ET in most reported cases, perhaps because of the suppression of the ET clone by both the cytotoxic effect of anti-myeloma therapy and the clonal repression by MM progression. PMID- 19129689 TI - Transfusion-associated iron overload as an adverse risk factor for transplantation outcome in patients undergoing reduced-intensity stem cell transplantation for myeloid malignancies. AB - Transfusion-associated iron overload could be an important risk factor in myeloablative hematopoietic stem cell transplantation. However, few studies have evaluated the effect of iron overload in reduced-intensity stem cell transplantation (RIST). We evaluated 38 patients with myeloid malignancies, 16 with and 22 without iron overload, who received RIST. We used pretransplant serum ferritin as a marker of iron overload. There was a positive correlation between the number of transfused packed red blood cells and pretransplant serum ferritin levels (21.5 units and 1,578.7 microg/l in the iron overload group vs. 12 units and 739.3 microg/l in the iron non-overload group; p <0.01). Engraftment day and chimerism analysis were not affected by iron overload (p = 0.71 and 0.47, respectively). There were no differences in treatment-related mortality (p = 0.94), veno-occlusive disease (p = 0.99), acute and chronic graft versus host disease (p = 0.58 and 0.99, respectively) according to iron overload. There was a significant difference in disease-free and overall survival (35.8 and 27% in the iron overload group vs. 80.6 and 54.6% in the iron non-overload group; p = 0.01 and 0.03, respectively). We conclude that transfusion-associated iron overload is an adverse risk factor in RIST for myeloid malignancies. The clinical outcomes according to iron overload in RIST were different in myeloablative hematopoietic stem cell transplantation. A serial follow-up of serum ferritin level and judicious iron chelation therapy will be needed to manage the side effect of iron overload in RIST and improve transplantation outcomes. PMID- 19129690 TI - Heparin-induced thrombocytopenia and phlegmasia cerulea dolens of the upper limb successfully treated with fondaparinux. PMID- 19129691 TI - Genetic inversions among hemophilia A patients from Macedonia and Bulgaria. PMID- 19129692 TI - Hydroxyurea and anagrelide combination therapy in patients with chronic myeloproliferative diseases resistant or intolerant to monotherapy. PMID- 19129693 TI - Angiotensin II upregulates RAGE expression on podocytes: role of AT2 receptors. AB - BACKGROUND: Advanced glycation end products (AGEs) play an important role in diabetic nephropathy. The receptor for AGEs, called RAGE, is present on podocytes. We investigated whether angiotensin II (ANG II) modulates RAGE expression on cultured differentiated podocytes. RESULTS: Cultured podocytes expressed AT1 and AT2 receptors. Surprisingly, ANG II induced RAGE mRNA and protein expression through AT2 receptors. ANG II had no influence on proliferation or protein content of podocytes. The increase in RAGE expression depended on stimulated transcriptional activity. Using various mutant reporter constructs of the RAGE promoter region, it was shown that a NF-kappaB binding site at -1519 was essential for ANG II-induced transcriptional activity. Preincubation with ANG II increased the expression of tumor necrosis factor-alpha mRNA and protein expression induced by AGE, indicating that the ANG II-mediated upregulation of RAGE has functional consequences. AGE-BSA was incorporated into cells as measured by Western blots for N epsilon-carboxymethyllysine, but ANG II did not influence this process. ANG II in the absence or presence of AGE-BSA did not induce apoptosis of podocytes. CONCLUSION: Our study revealed aninteraction between the renin-angiotensin system and the AGE/RAGE axis in podocytes. Since intraglomerular ANG II levels are increased in diabetic nephropathy, this interaction may have pathophysiological consequences for podocyte injury and inflammation associated with the development of diabetic nephropathy. PMID- 19129694 TI - Indoxyl sulfate inhibits nitric oxide production and cell viability by inducing oxidative stress in vascular endothelial cells. AB - BACKGROUND/AIM: Cardiovascular disease is a major cause of mortality in chronic kidney disease patients. Oxidative stress and nitric oxide (NO) deficiency play an important role in vascular endothelial cell dysfunction in chronic kidney disease. To determine if the uremic toxin indoxyl sulfate (IS) induces oxidative stress and inhibits NO production and cell viability in human umbilical vein endothelial cells (HUVEC). METHODS: The production of reactive oxygen species (ROS), superoxide, NO and peroxynitrite was measured using a fluorescence microplate reader. The expression of NADPH oxidases (Nox4, Nox2) was analyzed by quantitative reverse transcription-polymerase chain reaction. Cell viability was examined by 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate assay. RESULTS: IS induced ROS generation in HUVEC. An inhibitor of NADPH oxidase showed an inhibitory effect on IS-induced ROS production. However, the inhibitors of xanthine oxidase, mitochondrial electron transport and NO synthase did not show any significant effect on IS-induced ROS production. Antioxidants such as vitamin E, N-acetyl-L-cysteine and vitamin C inhibited IS induced ROS production. IS induced the expression of Nox4 mRNA and the production of superoxide and peroxynitrite in HUVEC. IS inhibited NO production in HUVEC. IS inhibited cell viability, and antioxidants preserve the inhibitory effect of IS on cell viability. CONCLUSIONS: IS inhibits NO production and cell viability by inducing ROS through induction of Nox4 in HUVEC. PMID- 19129695 TI - Fos expression in the rat vestibular nucleus complex after plugging the three semicircular canals. AB - HYPOTHESIS: Differential compensation after unilateral semicircular canal plugging (UCP) and unilateral labyrinthectomy (UL) may involve different expression patterns of fos mRNA and Fos protein in the vestibular nucleus complex. BACKGROUND: The expression of Fos protein in the vestibular nuclei underwent spatiotemporal changes related to the compensation process following surgical UL and chemical UL. These changes were temporally related to the appearance and development of vestibular compensation in the rat. METHODS: Spatiotemporal changes in both fos mRNA and protein levels in the rat brain after UCP were observed. The Fos expression pattern after UCP was compared to that after UL. RESULTS: Greater fos mRNA and Fos protein expressions were detected in bilateral medial vestibular nuclei (MVNs) after UL than after UCP. The changes were relatively long-lived after UL compared to those following UCP. CONCLUSION: Differences in the expression pattern of fos mRNA and Fos protein in bilateral MVNs between UCP and UL may explain differences in compensation after UCP and UL. PMID- 19129696 TI - A normative study of early prelingual auditory development. AB - The normal trajectory of early prelingual auditory development from birth to 24 months of age was characterized in a sample of 120 normal-hearing infants and toddlers of Mandarin-Chinese-speaking parents. The Infant-Toddler Meaningful Auditory Integration Scale (IT-MAIS) was administered to parents as a structured interview during routine pediatric health examinations. Developmental trajectories for the overall IT-MAIS score, as well as for scores on the subscales that assess behavioral evidence of spontaneous detection of and responsiveness to sound and of spontaneous recognition and discrimination of sound, were represented by logarithmic regression functions. On average, these regression functions characterized over 80% of the age-related variance in each scale. The developmental trajectories revealed that by 16 months the average infant exhibited spontaneous detection of and responsiveness to sound at all times, while spontaneous recognition and discrimination of sound was seen at all times after 26 months. The trajectory for the overall IT-MAIS scale, which combines the two subscales, reached ceiling at 22 months. The overall trajectory for Chinese infants closely matches the trajectory for normal-hearing infants of Arabic- and Hebrew-speaking parents in Israel. The Chinese IT-MAIS also exhibits similar internal consistency and item reliability to the German, Polish, and British English IT-MAIS scales. These similarities seen across culturally and linguistically diverse populations suggest the early prelingual auditory development follows the same, or a very similar course, in all infants. PMID- 19129697 TI - Audiometric characteristics of USH2a patients. AB - Temporal processing, frequency discrimination and frequency resolution, three basic mechanisms involved in speech perception, were studied in 11 USH2a patients from 10 Dutch families. Measurements included loudness scaling, gap detection, determination of auditory filter shapes and difference limen for frequency. The results were compared to values obtained from listeners with normal hearing, DFNA8/12 patients, DFNA13 patients and unselected patients with presbyacusis. Overall, the results for USH2a were most similar to those obtained in patients with sensorineural hearing loss caused by hair cell defects. To our knowledge, this is the first study that presents results on audiometric measurements combined with psychophysical measurements in a group of USH2a patients. PMID- 19129698 TI - Existence of possible functional interaction between the saccule and the posterior semicircular canal in humans: an evaluation using VEMPs. AB - The purpose of this study was to determine the effects of stimulation of the vertical semicircular canals on vestibular evoked myogenic potentials (VEMPs). VEMPs were recorded in 14 subjects seated with their heads and necks tilted 120 degrees forward on the interaural axis. The head was rotated 45 degrees to the left or right from the sagittal plane; an angular acceleration around the earth's vertical axis was then provided. When the posterior semicircular canal (PSCC) of the recording side was excited, the rotation was defined as 'ipsilateral rotation', and the opposite rotation was defined as 'contralateral rotation'. The background muscle activity-corrected p13-n23 amplitude (CA) in the ipsilateral rotation was significantly smaller than CAs in the static state and the contralateral rotation. Functional interaction between the saccule and the PSCC could be detected. PMID- 19129699 TI - Differential expression of matrix metalloproteinases and tissue inhibitors and extracellular matrix remodeling in aortic regurgitant hearts. AB - OBJECTIVES: Myocardial fibrosis in experimental aortic regurgitation (AR) features abnormal fibronectin with normal collagen content, but the relevant degradative processes have not been assessed. METHODS: To elucidate these degradative processes, mRNA (Northern) and protein levels (Western) of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), as well as MMP activity (zymography), were measured in cardiac fibroblasts (CF) from New Zealand white rabbits with experimental AR paired with normals (NL). Collagen and fibronectin were quantified by immunohistochemical staining. RESULTS: In AR CF versus NL CF, MMP-2 and -14 mRNA and protein were increased (both p < 0.005), while TIMPs 1-3 were slightly decreased (p < 0.05-0.005; TIMP-4 undetectable). Gelatinase activity in AR CF was 1.7 times that in NL CF (p < 0.005); fibronectinase activity was unaffected. The Jun N-terminal kinase (JNK) inhibitor SP600125 suppressed MMP-2 protein (0.4-fold, p < 0.05) and mRNA (0.7-fold, p < 0.005) in AR CF; MMP-2 levels in NL CF were unaffected. AR MMP-9 mRNA, protein and activity were low and indistinguishable from NL. In left ventricular tissue, fibronectin was increased 1.9-fold (AR vs. NL, p < 0.05). Total AR collagen was indistinguishable from NL, but the collagen III to collagen I isoform ratio decreased (0.4-fold, p < 0.05). CONCLUSIONS: Collagen is relatively deficient in AR fibrosis, due at least in part to upregulated MMPs and downregulated TIMPs; fibronectinase is unaltered. JNK-dependent regulation may stimulate both MMP-2 and fibronectin expression in AR, providing a potential therapeutic target. PMID- 19129701 TI - Does a reliable decline in Mini Mental State Examination total score predict dementia? Diagnostic accuracy of two reliable change indices. AB - BACKGROUND/AIM: Norms for change in Mini Mental State Examination (MMSE) total score suggest that only a decline of at least 2-4 points indicates a reliable change. However, it is unknown whether change norms (Reliable Change Indices, RCIs) of the MMSE total score are suitable to predict future dementia. METHODS: 554 elderly individuals aged 75 and over without dementia at the first 2 visits were tested with the MMSE at a maximum of 6 visits with 1.5-year intervals. Two different RCIs for change in MMSE score (first to second visit) were computed - one RCI which corrects for practice and one RCI which corrects for regression to the mean. The main outcome measure was the diagnosis of dementia. RESULTS: During the study, 88 persons developed dementia. RCIs were significantly associated with future dementia diagnosis. The best cutoff for raw change in MMSE total score to predict dementia was -1 point (sensitivity = 48%, specificity = 67%, relative risk = 1.6). With the RCI + regression to the mean, the diagnostic accuracy was moderate (sensitivity = 61%, specificity = 72%, relative risk = 3.2). CONCLUSION: A change in MMSE total score is significantly associated with future dementia, but the diagnostic accuracy for dementia prediction is rather low. PMID- 19129700 TI - Serum lipids are related to Alzheimer's pathology in nursing home residents. AB - BACKGROUND: Studies of associations between serum lipids and Alzheimer's disease (AD) or other dementias in the elderly show conflicting results, perhaps due to misclassification of the various dementias. METHODS: For 358 nursing home residents, serum lipids were studied at admission and diagnoses established at autopsy. We used defined neuropathological criteria to distinguish the presence of AD and to avoid errors of clinical dementia assessment. RESULTS: Residents with any AD pathology, as compared to those without AD pathology, had higher mean serum total cholesterol (TC; 200.4 vs. 185.9 mg/dl; p = 0.02) and higher mean low density lipoprotein cholesterol (LDL; 124.5 vs. 111.5 mg/dl; p = 0.03). Further, mean TC, LDL and high-density lipoprotein cholesterol levels all increased progressively with increasing pathological certainty of AD (p for trend = 0.001, 0.02 and 0.02). CONCLUSIONS: TC and LDL were significantly related to pathologically defined AD. If serum lipids have a role in the pathogenesis of AD, interventions may modify the course of disease. PMID- 19129702 TI - Lectin UEA-I-binding proteins are specifically increased in the squamous epithelium of patients with Barrett's esophagus. AB - BACKGROUND: Chronic gastritis and esophagitis are associated with changes in mucosal glycosylation patterns. Lectins represent a group of specific carbohydrate binding proteins that can be used as sensitive tools for the analysis of glycosylation patterns. AIM: To investigate the binding patterns of lectins Ulex europaeus agglutinin-I (UEA-I), Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA) and peanut agglutinin (PNA) at the gastroesophageal junction in nonerosive (NERD), erosive reflux disease (ERD) and Barrett's esophagus (BE). METHODS: One hundred and twenty-two patients (female n = 53; male n = 69; controls n = 28; NERD n = 36; ERD n = 24 and BE n = 34) were included in this study. The binding patterns of lectins were examined immunohistochemically at the squamocolumnar junction, in squamous epithelium and columnar-lined epithelium. Staining patterns of lectins were semiquantitatively evaluated using an immunohistochemical score and the data were analyzed using non parametric tests. RESULTS: BE, as compared to the controls, was associated with specific lectin-binding patterns: lectin binding of UEA-I and DBA were significantly decreased at the superficial (p = 0.012; p = 0.00036, respectively) and at the deep glandular body (p = 0.045; p = 0.055, respectively). Comparisons of lectin-staining scores between GERD and BE revealed significant increases of UEA-I in both the stratum superficiale (p = 0.0155) and stratum spinosum (p = 0.0048) of SE in patients with BE. Notable, this change was specific for patients with BE, while no difference was observed between patients with GERD and controls. CONCLUSION: We found two major types of lectin-binding patterns. First, lectin-binding characteristics associated with GERD in general, and second, lectin-binding patterns which were specific for BE. Lectin UEA-I-binding proteins were specifically increased in the squamous epithelium of patients with BE. Thus, UEA-I may serve as a potential marker for BE, especially in patients with short segment BE. PMID- 19129703 TI - Parkinson's disease with and without REM sleep behaviour disorder: are there any clinical differences? AB - Rapid eye movement sleep behaviour disorder (RBD) may serve as a useful indicator to approach Parkinson's disease (PD); however, PD patients do not always exhibit RBD. We wondered whether the presence of RBD would be reflected in the expansion of PD lesions and represent the same PD entity. We examined the clinical differences between PD with and without RBD and studied the frequency of RBD-like symptoms (RBD-s) and clinical differences in 150 PD patients, including 81 patients (54.0%) who satisfied the International Classification of Sleep Disorders, Revised, minimum clinical criteria for RBD. RBD-s preceding the appearance of parkinsonism were found in 44.4% of patients. Statistically, the presence of RBD-s was associated with ages above 65 years, male gender, constipation, dopa-induced dyskinesia and 'sleep attack', with odds ratios of 3.709, 2.469, 2.184, 5.046 and 6.562, respectively. No differences were found between the 2 groups with regard to symptoms at PD onset, disease duration, Hoehn Yahr stage, hallucination, dementia, wearing-off, orthostatic hypotension, cerebral blood flow and antiparkinsonism drugs. In the early stage, RBD and autonomic system dysfunction are important factors in the progression of PD. PMID- 19129704 TI - A prospective study of alterations in balance among patients with Parkinson's Disease. Protocol of the postural evaluation. AB - The aim of this study is to determine whether the balance problems experienced by Parkinson's disease (PD) patients may in part be due to dysfunctional processing of vestibular information, and to search for factors that may help predict the risk of falls. We evaluated the balance of 45 idiopathic PD patients and 20 healthy subjects by means of computerized dynamic posturography using sensory organization tests (SOT), rhythmic weight shift (RWS) tests and limits of stability (LOS) tests; and by the timed up-and-go (TUG) test. PD patients had poorer scores in the SOT than controls for overall balance and vestibular and visual inputs. They also performed worse in RWS and LOS tests, and were slower in performing the TUG test. Hoehn-Yahr stage did not correlate with vestibular input. Balance impairment in PD patients involves deteriorated processing of vestibular input, but this deterioration is independent of disease progression. Falls are related to PD patients' reduced limits of stability. PMID- 19129705 TI - Switching to second-line therapies in interferon-beta-treated relapsing-remitting multiple sclerosis patients. AB - BACKGROUND: Interferon-beta (IFNB) therapies are the most widely used as first line intervention in the treatment of relapsing-remitting (RR) multiple sclerosis (MS). Despite long-term experience, however, the definition and prediction of response remain controversial. AIM: The objective of this study was to assess the long-term validity of the main clinical definitions of response applied after 1 and 2 years of IFNB therapy in a cohort of RRMS patients followed up for at least 5 years. METHODS: We tested these different definitions against a 'hard' parameter of treatment failure, represented by the need to suspend IFNB and switch to an intravenous immunosuppressive (IVIS) treatment, using Kaplan-Meier and Cox survival analyses. RESULTS: Out of 147 RRMS patients treated with IFNB therapy and followed up for 7.8 +/- 2.1 years, 26 (18%) were switched to an IVIS therapy. On the whole, disability progression as indicated using the Expanded Disability Status Scale (EDSS) and a higher number of relapses in the first 2 years of therapy were related to long-term treatment failure. CONCLUSION: Our study highlights the role of disability and high relapse rate in the first 2 years of treatment in predicting long-term response and the switching to second line therapies. PMID- 19129706 TI - Broca's aphasiacs. AB - After Gall, Bouillaud and Auburtin had localized the function of language to the frontal lobes in the early 19th century, Paul Broca's famous patient, M. Leborgne (known as 'Tan'), was described to the Anthropological Society of Paris and his case was published in the Bulletin de la Societe Anatomique, in 1861. Broca relied on the uncut brain for his clinicopathological inferences. A few months later, his second case, M. Lelong, yielded similar pathological details and confirmed Broca's localization of language. The subsequent controversies with Dax and Pierre Marie are summarized. More recent imaging of the brains of Lelong and Leborgne has partly vindicated Broca's controversial conclusions. Most papers on Broca's work contain only brief, derivative references to his 1861 paper; the actual contents, translated into English, are reproduced here. PMID- 19129707 TI - Craniorachischisis totalis: a case report and review of the literature. AB - OBJECTIVE: To discuss a fetus with craniorachischisis diagnosed antenatally and to review the literature. METHOD: Case report. RESULTS: Craniorachischisis was detected on ultrasound scan in a fetus at gestational week 13. Pregnancy was terminated and diagnosis was verified postnatally. CONCLUSION: Craniorachischisis is a rare and severe form of neural tube defects (NTDs). The majority of currently known cases of mouse craniorachischisis have been found to result from disturbance of a single molecular signaling cascade, called planar cell polarity pathway (PCP). The mutant genes that have been causative in disturbance of PCP in mouse models have been examined in human malformations but none of them have so far been implicated in human craniorachischisis. To date, no other genes except the gene encoding 5,10-methylenetetrahydrofolate reductase have been specifically implicated in predisposition to NTDs. We suggest that other PCP genes should be considered as candidates for a role in the etiology of human NTDs. Further investigations are therefore necessary. PMID- 19129708 TI - Percutaneous laser ablation of sacrococcygeal teratoma in a hydropic fetus with severe heart failure--too late for a surgical procedure? AB - Sacrococcygeal teratoma (SCT) is the commonest solid fetal tumor. Perinatal prognosis is usually favorable, but sometimes it can be complicated by fetal hydrops being responsible for high risk of mortality. Fetal therapy in such cases has so far not been established. We report a case with a giant solid SCT associated with fetal hydrops and severe heart failure. 2D- and 3D-Doppler ultrasonography revealed great vessels originated from the medial sacral artery. Percutaneous laser ablation of these vessels was performed at 24 weeks of gestation. During the procedure, severe anemia was also diagnosed (hemoglobin 4.3 g/dl). Two days later, the fetus died and pathological examination revealed local tumor necrosis and blood hemorrhage inside the mass. We suggest that in such cases, fetal surgery may not be enough, being too late, and perhaps fetal clinical therapy for anemia and heart failure could be the best option at a gestational age of less than 28 weeks. PMID- 19129709 TI - Aerodynamics of the pseudo-glottis. AB - OBJECTIVE: The aim of this work is to study the hitherto unclear aerodynamic parameters of the pseudo-glottis following total laryngectomy. These parameters include airflow rate, sub-pseudo-glottic pressure (SubPsG), efficiency and resistance, as well as sound pressure level (SPL). PATIENTS AND METHODS: Eighteen male patients who have undergone total laryngectomy, with an age range from 54 to 72 years, were investigated in this study. All tested patients were fluent esophageal 'voice' speakers utilizing tracheo-esophageal prosthesis. The airflow rate, SubPsG and SPL were measured. RESULTS: The results showed that the mean value of the airflow rate was 53 ml/s, the SubPsG pressure was 13 cm H(2)O, while the SPL was 66 dB. The normative data obtained from the true glottis in healthy age-matched subjects are 89 ml/s, 7.9 cm H(2)O and 70 dB, respectively. Other aerodynamic indices were calculated and compared to the data obtained from the true glottis. CONCLUSION: Such a comparison of the pseudo-glottic aerodynamic data to the data of the true glottis gives an insight into the mechanism of action of the pseudo-glottis. The data obtained suggests possible clinical applications in pseudo-voice training. PMID- 19129710 TI - Insulin resistance markers in children. AB - The prevalence of obesity among children and adolescents is progressively increasing around the world. One of the important consequences of obesity is the development of insulin resistance (IR). This condition has a multifactorial pathogenesis and is associated with cardiovascular risk, diabetes, hypertension, polycystic-ovary syndrome and a shorter lifespan. IR during childhood may be diagnosed by physical examination or there may be clues in the histories of the patient and his/her family. When IR is suspected, tests on a blood sample (which are more reliable) are recommended. Most of the biochemical markers have been well defined in adults, but appropriate reference data for children are still lacking. Here we discuss the usefulness of various currently known biochemical markers to evaluate insulin sensitivity (homeostatic model assessment, the quantitative insulin sensitivity check index, the oral glucose tolerance test, Matsuda method and the whole-body insulin resistance index), hormones (leptin, adiponectin, resistin, glucocorticoids, the insulin-like growth factor-1-binding protein/growth hormone axis, ghrelin, sex hormone-binding globulin and retinol binding protein-4) and inflammatory markers (C-reactive protein, IL-6, intercellular adhesion molecule-1, vascular adhesion molecule-1 and E-selectin), which can be used in the diagnosis of IR in children. PMID- 19129711 TI - Inactivating mutations of luteinizing hormone beta-subunit or luteinizing hormone receptor cause oligo-amenorrhea and infertility in women. AB - Women harbouring inactivating mutations in luteinizing hormone (LH) beta subunit (LHB) or LH receptor (LHCGR) genes have similar clinical manifestations characterized by female external genitalia, spontaneous breast and pubic hair development at puberty, and normal or late menarche followed by oligo-amenorrhea and infertility. Oestradiol and progesterone levels are normal for the early to midfollicular phase, but do not reach ovulatory or luteal phase levels, confirming lack of ovulation. Notably, serum LH levels are low in patients with LHB mutations and high in those with LHCGR mutations, whereas follicle stimulating hormone levels are normal or only slightly increased. Pelvic ultrasound has demonstrated a small or normal uterus and normal or enlarged ovaries with cysts. Women with LHB mutations may be treated with hCG (human chorionic gonadotropin) or LH, whereas those with mutations in LHCGR are resistant. Lhb and Lhcgr knockout female mice are close phenocopies of the respective human mutations, and confirm that early follicular development, low levels of oestrogen production and theca cell development are independent of LH action, which is necessary for ovulation. Although inactivating mutations in LHB and LHCGR are rare in comparison to other genetic and non-genetic causes of hypogonadism, they should be considered in the differential diagnosis of oligo amenorrhea and infertility. PMID- 19129712 TI - Triglycerides and alanine aminotransferase as screening markers for suspected fatty liver disease in obese children and adolescents. AB - BACKGROUND/AIMS: Metabolic syndrome (MS) and fatty liver disease (FLD) are on the rise. The association between these conditions in Brazilian youth is analyzed. METHODS: 354 subjects (11.2+/-3.1 years) were evaluated. FLD was suspected by ultrasound and computed tomography; weight and MS by BMI z-score and NCEP-ATPIII respectively. RESULTS: Subjects were classified as: group 1 with suspected FLD and group 2 without and group 2 as 'a' (overweight/obese) and 'b' (normal weight). Comparing group 1 with 2a and 2b, differences in age (p=0.016; p=0.075), triglycerides (TG) (p=0.021; p=0.002), insulin (p=0.652; p=0.015) and homeostasis model assessment method of IR (HOMA-IR) (p=0.737; p=0.003) were found. Group 2a was divided into low/high alanine aminotransferase (ALT). A decrease in waist circumference and TG was found going from those with suspicion of FLD to obese with high and low ALT. Insulin and HOMA-IR in group 1 and high ALT were similar. Gender (OR 6.6; CI 1.9-22.5; p=0.025), age (OR 1.3; CI 1.1-1.6; p=0.006), TG (OR 10.4; CI 3.1-34.4; p=0.005) were associated with suspected FLD. For every 10 U/l increase in ALT, there was a 4-fold greater chance of probable FLD (OR 4.01; CI 2.06-9.40; p<0.001). CONCLUSION: Measurements of ALT and TG should be considered as screening for suspected FLD in overweight/obese youth. PMID- 19129713 TI - Expression and activity of 11beta-hydroxysteroid dehydrogenase type 1 enzyme in subcutaneous and visceral adipose tissue of prepubertal children. AB - BACKGROUND: Glucocorticoid excess promotes visceral obesity and cardiovascular disease. Ligand availability to the glucocorticoid receptor is controlled by isoforms of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) which converts endogenous cortisone to active cortisol. AIM: To evaluate the expression and activity of 11beta-HSD1 in subcutaneous adipose tissue (SC) and visceral adipose tissue (VAT) in prepubertal children with normal weight. METHODS: Fourteen patients (11 female/3 male) with a mean age of 6.9+/-0.9 years and a body mass index (BMI) of 17.4+/-0.61 underwent elective open abdominal surgery. RESULTS: Expression of 11beta-HSD1 mRNA in SC and VAT was similar (0.8+/-0.15 vs. 0.61+/ 0.12 AU). The activity of this enzyme in SC was significantly lower compared to VAT (1.42+/-0.39 vs. 2.79+/-0.61 ng cortisol/g tissue/24 h, p<0.05). In addition, we observed a significant direct correlation with the expression of 11beta-HSD1 in VAT adipose tissue with the patient's BMI (r=0.825, p=0.002). CONCLUSIONS: This correlation together with the increased activity of this enzyme in visceral adipose tissue might contribute to decreased hepatic insulin sensitivity due to increased portal cortisol when BMI increases. These observations appear to be particularly important in children born with low birth weight who develop rapid early weight gain. PMID- 19129714 TI - Inhibin A production after gonadotropin stimulus: a new method to detect ovarian tissue in ovotesticular disorder of sex development. AB - BACKGROUND/AIMS: While laboratory methods for the detection of testicular tissue are well standardized, currently there is no available test to demonstrate the presence of ovarian tissue. We evaluated the effectiveness of gonadal stimulation with luteinizing hormone (LH)/follicle-stimulating hormone (FSH) for the detection of ovarian tissue in patients with disorders of sex development (DSD). METHODS: Ten patients with congenital adrenal hyperplasia (CAH) as ovarian positive controls, 10 with cryptorchidism (ovarian-negative controls), 13 patients with DSD of no defined etiology and 7 patients with ovotesticular DSD (true hermaphroditism, TH) were included in the study. They underwent a daily injection of both LH and FSH on 3 consecutive days. LH, FSH, estradiol, testosterone and inhibin A were measured before treatment, 24 h after the 1st dose and 24 h after the 3rd dose. RESULTS: Estradiol increased in all CAH and TH patients, with a median value of 155.1 and 92.6 pg/ml, respectively, after the 3rd injection. Inhibin A also increased in all CAH and TH patients, with a median value of 70.4 and 32.2 pg/ml, respectively, after the 3rd injection. There was no change in these hormones in the other groups. CONCLUSION: The LH/FSH stimulation test might be a useful method to detect the presence of ovarian tissue. PMID- 19129716 TI - Autosomal dominant neurohypophyseal diabetes insipidus in two families. Molecular analysis of the vasopressin-neurophysin II gene and functional studies of three missense mutations. AB - BACKGROUND: Autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is a rare disease with symptoms of polydipsia, polyuria and dehydration caused by arginine vasopressin deficiency. Disease onset is within infancy or adolescence. A variety of disease-causing mutations of the arginine vasopressin neurophysin II gene (AVP) on chromosome 20p13 have been described. METHODS: Two Polish families with adFNDI were screened for mutations. Processing of wild-type (WT) and mutant AVP was monitored using immunocytochemical methods in stably transfected Neuro2A cells. AVP secretion into the cell culture supernatant was investigated with an enzyme immunoassay. RESULTS: In the first family a heterozygous p.G96D mutation was identified. Some patients additionally carried a novel heterozygous mutation p.A159T. The second family presented with a heterozygous mutation p.C98G. Confocal laser microscopy unveiled accumulation of p.G96D and p.C98G prohormones in the cellular bodies, whereas WT and p.A159T prohormones and/or processed products were located in the tips of cellular processes. Reduced levels of AVP in supernatant culture medium of p.G96D and p.C98G transfected cells in comparison to p.A159T and WT cells were found. CONCLUSIONS: We conclude that the p.G96D and p.C98G mutations cause adFNDI in the two reported families. The sequence variant p.A159T does not seem to have disease causing effects. PMID- 19129715 TI - Three novel IGFALS gene mutations resulting in total ALS and severe circulating IGF-I/IGFBP-3 deficiency in children of different ethnic origins. AB - BACKGROUND/AIMS: To date, four mutations in the IGFALS gene have been reported. We now describe two children of different ethnic background with total acid labile subunit (ALS) and severe circulating IGF-I/IGFBP-3 deficiencies resulting from three novel mutations in the IGFALS gene. PATIENTS/METHODS: Serum and DNA of patients were analyzed. RESULTS: Case 1 is a 12-year-old boy of Mayan origin. Case 2 is a 5-year-old girl of Jewish/Eastern European (Polish, Russian, Austrian Hungarian)/Icelandic/European (French, English) ancestry. The reported cases had moderate short stature (-2.91 and -2.14 SDS, respectively), nondetectable serum ALS and extremely low serum concentrations of IGF-I and IGFBP-3. Case 1 harbored a novel homozygous 1308_1316 dup9 mutation in a highly conserved leucine-rich repeat (LRR) 17 motif of exon 2, representing an in-frame insertion of 3 amino acids, LEL. Case 2 harbored a novel heterozygous C60S/L244F mutation in exon 2, located within a highly conserved LRR 1 and LRR 9, respectively. CONCLUSIONS: The identification of additional novel IGFALS mutations, resulting in severe IGF I/IGFBP-3 and ALS deficiencies, supports IGFALS as a candidate gene of the GH/IGF system, implicated in the pathogenesis of primary IGF deficiency, and represents an important part of its differential diagnosis. PMID- 19129717 TI - X-linked adrenal hypoplasia congenita caused by a novel intronic mutation of the DAX-1 gene. AB - OBJECTIVE: X-linked adrenal hypoplasia congenita (AHC) associated with hypogonadotropic hypogonadism (HHG) is caused by mutations of the DAX-1 gene. A novel intronic mutation of the gene and the results of in vitro expression analysis are reported. PATIENT AND METHODS: The patient is a 13-year-old boy who presented with severe dehydration and salt loss in the neonatal period, when he lacked cutaneous pigmentation and elevation of plasma ACTH level. He had been diagnosed and treated as adrenal hypoplasia. The DAX-1 gene was analyzed by direct DNA sequencing in the patient and his parents. In vitro expression analysis was applied to confirm the consequent missplicing of mRNA caused by the mutation. RESULTS: The substitution of guanine for cytosine at the splice donor site (IVS1 + 1G>C) was observed in our patient and his mother. Expression analysis suggested that this mutation may hinder splicing of the adjacent intron and cause alternative activation of three cryptic splice donor sites within exon 1 leading to the aberrant splicing of mRNA and production of truncated proteins. CONCLUSION: To our knowledge, this is the first case report of AHC associated with HHG caused by an intronic mutation of the DAX-1 gene. The clinical course of our patient may suggest that the onset of mineralocorticoid deficiency can precede that of glucocorticoid deficiency in some patients with AHC, and thus appear to suffer from aldosterone synthase deficiency at the onset of the disease. PMID- 19129719 TI - Effects of changes in alveolar ventilation on isoflurane arterial blood concentration and its uptake into the human body. AB - We investigated whether minute alveolar ventilation affects isoflurane concentration in arterial blood and uptake of isoflurane into the body. Thirty female patients scheduled to undergo elective gynecological surgery were randomly assigned to one of three groups: i.e. hyperventilation, normal ventilation and hypoventilation. Inspiratory (CIiso) and end-tidal (CEiso) concentrations of isoflurane were measured by infrared analysis, and arterial blood isoflurane concentration (Aiso) was analyzed by gas chromatography. Cardiac index was measured by Doppler ultrasonography. The body uptake of isoflurane was determined by multiplying alveolar ventilation by the gradient of CIiso-CEiso. Aiso was highest in the hyperventilation group (significant), followed by the normal ventilation and hypoventilation groups, during the 40-min study. During the first 10 min of the study, the slope of the Aiso-over-time curve was highest in the hyperventilation group, followed by the normal ventilation group and the hypoventilation group. During the second half of the study (20-40 min), the slope Aiso-over-time curve did not differ among the three groups. Changes in ventilation affected the concentration of isoflurane in arterial blood but did not significantly alter the uptake of it during the last 20 min of the study. The change of alveolar ventilation altered the speed of functional residual capacity wash-in by isoflurane, which was the integral factor influencing Aiso and body uptake of isoflurane. PMID- 19129718 TI - Correlation of Fc(gamma)RIIa (CD32) Polymorphism and IgG Antibody Subclasses in Hemolytic Disease of Newborn. AB - ABO-HDN is a common disease of newborn in China and currently there is no satisfactory method to predict it in the antepartum period. It has been reported that Fc(gamma)RIIa (CD32) genotype is associated with both infectious diseases induced by bacteria and parasitemia. There is a relationship between IgG subclass and RH-HDN. To study the pathogenesis of ABO-HDN and to find reliable method to diagnose ABO-HDN, we investigated the polymorphism of Fc(gamma)RIIa (CD32) and distribution of IgG subclass in infants with ABO-HDN and their mothers by polymerase chain reaction or ELISA assay. We observed that the frequency of HH131 genotype is lower in infants with ABO-HDN than in controls (p < 0.01), while the frequency of HR131 genotype is higher in ABO-HDN infants than that in controls (p < 0.01). The genotype HR131 and concentrations of IgG1 and IgG3 are significantly correlated with ABO-HDN. Our results demonstrated, for the first time, that there is a correlation between ABO-HDN and CD32, and different IgG subclass distribution. Our study may contribute to the development of an early diagnostic method for HDN. PMID- 19129727 TI - Endotoxinemia-induced changes in coagulation as measured by rotation thrombelastometry technique and conventional laboratory tests: results of a pilot study on pigs. AB - Modified rotation thrombelastometry (ROTEM) is widely used in near-patient assessment of hemostasis, but data on functional consequences initiated by acute endotoxinemia are rare. To test the hypothesis that the ROTEM technique allows detection of endotoxinemia-induced changes in hemostasis, we conducted a pilot study on pigs. Fifteen healthy pigs were anesthetized and instrumented for invasive hemodynamic monitoring. Several coagulation tests and the ROTEM assay were performed at baseline and 60 min after administration of a bolus of 200 microg of Escherichia coli lipopolysaccharide followed by a continuous infusion of 0.1 microg/kg per min. After induction of acute endotoxinemia, clot formation time increased (P = 0.001), and alpha angle (P = 0.001) and maximum clot firmness decreased significantly (P = 0.001) in intrinsically and extrinsically activated ROTEM assays. Moreover, fibrinogen/fibrin polymerization showed significantly lower values during endotoxinemia (P = 0.001), and coagulation time shortened for the intrinsically activated assay (P = 0.017). Simultaneously, a significant decrease in platelet count (P = 0.001), fibrinogen (P = 0.001), antithrombin and protein C (P = 0.001) was registered, whereas results of standard coagulation tests and D-dimers showed no significant changes although thrombin-antithrombin complex increased (P = 0.001). Wilcoxon Z score analysis showed that changes in ROTEM variables were comparable to changes in antithrombin, protein C, platelet count, white blood cells and fibrinogen concentrations. The ROTEM assays were able to reflect endotoxinemia-dependent changes in the hemostatic system in pigs early by showing not only activation but also signs of consumption, whereas results of routine coagulation tests remained unchanged. PMID- 19129726 TI - Heritability of plasma concentrations of activated protein C in a Spanish population. AB - The protein C anticoagulant pathway plays a crucial role in the regulation of fibrin formation. Protein C is activated on the surface of endothelial cells by the thrombin-thrombomodulin complex with the stimulation of the endothelial protein C receptor. The levels of circulating activated protein C reflect in-vivo protein C activation, and a low level of activated protein C is a risk factor for venous thromboembolism. The objective of the study was to assess the relative contributions of genetic and environmental factors to the variation in the levels of activated protein C and protein C. Blood samples were collected from 126 individuals belonging to 19 Spanish families, and heritability and common household effect were estimated for protein C, activated protein C and its complexes with protein C and alpha1-antitrypsin. In addition, we calculated the genetic correlation between protein C and activated protein C phenotypes. Although all phenotypes showed significant heritability, activated protein C phenotype resulted in a very high heritability of 83%, which clearly shows that this phenotype is strongly influenced by the action of gene(s). Furthermore, the bivariant analyses of protein C and activated protein C phenotypes indicate that there is a high genetic correlation between them (0.74). Nevertheless, this correlation is counteracted by a negative environmental correlation (-0.54) resulting in a phenotypic correlation of 0.35. The presence of such strong genetic effects suggests that it will be possible to localize the loci that influence this phenotype and determine the contribution to the risk of thrombosis. PMID- 19129728 TI - The embolic storm: dramatic peripheral complications of left ventricular thrombosis after myocardial infarction. AB - Left ventricular thrombus is a common complication after acute myocardial infarction. Although anticoagulant treatment has an established role in reducing the rate of thrombus formation and embolic phenomena during acute myocardial infarction, prior studies showed that left ventricular thrombus and systemic embolization may develop irrespective of adequate anticoagulant therapy. We present a descriptive case of a patient with left ventricular thrombus who experienced dramatic peripheral embolic events soon after the onset of anterior myocardial infarction despite full anticoagulation, and we discuss possible pathogenesis. PMID- 19129731 TI - Short-term weather variability in Chicago and hospitalizations for Kawasaki disease. AB - BACKGROUND: : Kawasaki disease exhibits a distinct seasonality, and short-term changes in weather may affect its occurrence. METHODS: : To investigate the effects of weather variability on the occurrence of this syndrome, we conducted a time-between-events analysis of consecutive admissions for Kawasaki disease to a large pediatric hospital in Chicago. We used gamma regression to model the times between admissions. This is a novel application of gamma regression to model the time between admissions as a function of subject-specific covariates. RESULTS: : We recorded 723 admissions in the 18-year (1986-2003) study period, of which 700 had complete data for analysis. Admissions for Kawasaki disease in Chicago were seasonal: The mean time between admissions was 34% shorter (relative time = 0.66, 95% confidence interval 0.54-0.81) from January-March than from July-September. In 1998, we recorded a larger number of admissions for Kawasaki disease (n = 65) than in other years (mean n = 37). January-March months of 1998 were warmer by a mean of 3 degrees C (1.5 degrees C-4.4 degrees C) and the mean time between admissions was 48% shorter (relative time = 0.52, 0.36-0.75) than in equivalent periods of other study years. CONCLUSIONS: : Our findings show that atypical changes in weather affect the occurrence of Kawasaki disease and are compatible with a link to an infectious trigger. The analysis of interevent times using gamma regression is an alternative to Poisson regression in modeling a time series of sparse daily counts. PMID- 19129732 TI - Pranlukast attenuates ischemia-like injury in endothelial cells via inhibiting reactive oxygen species production and nuclear factor-kappaB activation. AB - The anti-inflammatory effects of pranlukast, an antagonist of cysteinyl leukotriene receptor 1, may be rendered not only by antileukotriene activity but also by other pharmacological activities. Previous studies indicate that pranlukast reduces ischemic tissue injury partially through decreasing vascular permeability, but its effect on ischemic injury in endothelial cells is not known. Thus, in this study, we investigated the effect of pranlukast on ischemia like injury induced by oxygen-glucose deprivation (OGD) in EA.hy926 cells, a human endothelial cell line, and the possible mechanisms. We found that cell viability was reduced, lactate dehydrogenase release was increased 4-8 hours after OGD, and necrosis was induced 8 hours after OGD. Production of reactive oxygen species (ROS) increased by 211%, 176%, and 128%, respectively, 0.5, 1, and 2 hours after OGD. Nuclear factor-kappaB (NF-kappaB) was translocated to the nuclei 4-8 hours after OGD. Pranlukast ameliorated the reduced viability, the increased lactate dehydrogenase release, and necrosis after OGD. It also reduced ROS production and inhibited NF-kappaB nuclear translocation after OGD. The ROS scavenger, edaravone, inhibited OGD-induced nuclear translocation of NF-kappaB as well. Edaravone and pyrrolidine dithiocarbamate (a specific NF-kappaB inhibitor) protected endothelial cells from the OGD-induced injury. However, zileuton, a 5 lipoxygenase inhibitor, did not affect the cell injury, ROS production, and NF kappaB nuclear translocation after OGD. The exogenous leukotriene D4 did not induce cell injury, ROS production, and NF-kappaB translocation. Thus, we conclude that pranlukast protects endothelial cells from ischemia-like injury via decreasing ROS production and inhibiting NF-kappaB activation, which is leukotriene independent. PMID- 19129730 TI - Fine particulate matter air pollution, proximity to traffic, and aortic atherosclerosis. AB - BACKGROUND: The initiation and acceleration of atherosclerosis is hypothesized as a physiologic mechanism underlying associations between air pollution and cardiovascular effects. Despite toxicologic evidence, epidemiologic data are limited. METHODS: In this cross-sectional analysis we investigated exposure to fine particulate matter (PM2.5) and residential proximity to major roads in relation to abdominal aortic calcification, a sensitive indicator of systemic atherosclerosis. Aortic calcification was measured by computed tomography among 1147 persons, in 5 US metropolitan areas, enrolled in the Multi-Ethnic Study of Atherosclerosis. The presence and quantity of aortic calcification were modeled using relative risk regression and linear regression, respectively, with adjustment for potential confounders. RESULTS: We observed a slightly elevated risk of aortic calcification (RR = 1.06; 95% confidence interval = 0.96-1.16) with a 10 microg/m contrast in PM2.5. The PM2.5-associated risk of aortic calcification was stronger among participants with long-term residence near a PM2.5 monitor (RR = 1.11; 1.00-1.24) and among participants not recently employed outside the home (RR = 1.10; 1.00-1.22). PM2.5 was not associated with an increase in the quantity of aortic calcification (Agatston score) and no roadway proximity effects were noted. There was indication of PM2.5 effect modification by lipid-lowering medication use, with greater effects among users, and PM2.5 associations were observed most consistently among Hispanics. CONCLUSIONS: Although we did not find persuasive associations across our full study population, associations were stronger among participants with less exposure misclassification. These findings support the hypothesis of a relationship between particulate air pollution and systemic atherosclerosis. PMID- 19129733 TI - Nitric oxide and catalase-sensitive relaxation by scutellarin in the mouse thoracic aorta. AB - The vascular activity of scutellarin (SCU), a flavonoid isolated from a Chinese traditional medicinal plant, was investigated in isolated thoracic aortic rings of mice. SCU-induced dose-dependent relaxation of phenylephrine (1 microM) stimulated contractions. This relaxation was reduced by endothelium removal, significantly reduced by both the nitric oxide synthase inhibitor (Nomega-nitro-L arginine methylester, 300 microM) and slightly limited by the soluble guanylyl cyclase inhibitor (1 H-[1,2,4] oxidazolol [4,3-a] quinoxalin-1-one, 100 microM). The catalase inhibitor (3-amino-1,2,4-triazole, 50 mM) augmented the constriction and blocked the lowest SCU concentration relaxation, whereas catalase addition was without effect. Preincubation with 300 and 1000 microM SCU significantly suppressed the contractile dose-response to phenylephrine, causing both a significant rise in half maximal effective concentration and a decrease in the maximal developed force. Western blot analysis showed that SCU inhibition of contraction was independent of reductions in myosin light chain phosphorylation. These results suggested that SCU relaxation was predominantly endothelium dependent and likely involved the catalase-sensitive nitric oxide synthase signaling pathway, without loss of myosin phosphorylation. The potential clinical use of SCU may prove to be effective in increasing vasoreactivity, independently of smooth muscle contractile activity that is mediated by the 20-kDa myosin light chain phosphorylation. PMID- 19129734 TI - Varespladib (A-002), a secretory phospholipase A2 inhibitor, reduces atherosclerosis and aneurysm formation in ApoE-/- mice. AB - The family of secretory phospholipase A2 (sPLA2) enzymes has been associated with inflammatory diseases and tissue injury including atherosclerosis. A-001 is a novel inhibitor of sPLA2 enzymes discovered by structure-based drug design, and A 002 is the orally bioavailable prodrug currently in clinical development. A-001 inhibited human and mouse sPLA2 group IIA, V, and X enzymes with IC50 values in the low nM range. A-002 (1 mg/kg) led to high serum levels of A-001 and inhibited PLA2 activity in transgenic mice overexpressing human sPLA2 group IIA in C57BL/6J background. In addition, the effects of A-002 on atherosclerosis in 2 ApoE mouse models were evaluated using en face analysis. (1) In a high-fat diet model, A-002 (30 and 90 mg/kg twice a day for 16 weeks) reduced aortic atherosclerosis by 50% (P < 0.05). Plasma total cholesterol was decreased (P < 0.05) by 1 month and remained lowered throughout the study. (2) In an accelerated atherosclerosis model, with angiotensin II-induced aortic lesions and aneurysms, A-002 (30 mg/kg twice a day) reduced aortic atherosclerosis by approximately 40% (P < 0.05) and attenuated aneurysm formation (P = 0.0096). Thus, A-002 was effective at significantly decreasing total cholesterol, atherogenesis, and aneurysm formation in these 2 ApoE mouse models. PMID- 19129735 TI - Sera from patients with diabetes do not alter the effect of mammalian target of rapamycin inhibition on smooth muscle cell proliferation. AB - Clinical studies of drug-eluting stents delivering the mammalian target of rapamycin (mTOR) inhibitor, rapamycin (Sirolimus), have demonstrated a reduced efficacy for these devices in patients with diabetes, which suggests that the mTOR pathway may cease to be dominant in mediating the vascular response to injury under diabetic conditions. We hypothesized that changes in serum composition accompanying diabetes may reduce the role of mTOR in mediating the vascular response to injury. We measured the ability of a median dose of rapamycin (10 nM) to inhibit the proliferation of human coronary artery smooth muscle cells (huCASMCs) stimulated with serum obtained from donors with diabetes (n = 14) and without diabetes (n = 16). In an additional analysis, we compared the effects of rapamycin on huCASMCs stimulated with the serum of donors with metabolic syndrome (n = 15) versus those without (n = 7). There was no difference in the effect of rapamycin on huCASMC proliferation after stimulation with serum from either donors with diabetes or donors with metabolic syndrome compared with the respective controls. We conclude that the changes in the serum composition common to diabetes and metabolic syndrome are insufficient to diminish the role of mTOR in the progression of cardiovascular disease. PMID- 19129736 TI - Hemodynamic and cardiac neurotransmitter-releasing effects in conscious dogs of attention- and wake-promoting agents: a comparison of d-amphetamine, atomoxetine, modafinil, and a novel quinazolinone H3 inverse agonist. AB - Conscious coronary sinus-cannulated dogs were used to assess the hemodynamic effects and local cardiac norepinephrine (NE) and histamine (HA) release of 4 mechanistically diverse agents either clinically approved or representing a potential novel mechanism for the promotion of wakefulness or attention. Dosing regimens were based on reported or concurrently determined wake-promoting activities in canine models. The central nervous system stimulant, d-amphetamine [0.1 mg x kg(-1) x 10 min intravenous (IV)], significantly elevated mean arterial pressure (+30%) and increased coronary sinus and peripheral venous NE concentrations, indicative of cardiac neurotransmitter release. The selective NE reuptake inhibitor atomoxetine (2.0 mg x kg(-1) x 10 min(-1) IV) and modafinil (30.0 mg x kg(-1) x 10 min(-1) IV) also significantly elevated mean arterial pressure (+15% and +30%, respectively), but with no effect on coronary sinus or peripheral NE concentration, suggesting central mechanisms underlying the hemodynamic effects. The preclinical demonstrations of pressor effects with d amphetamine, atomoxetine, and modafinil are consistent with clinically reported hemodynamic effects with these agents. The quinazolinone HA receptor subtype H3 inverse agonist 5r (0.3 mg x kg(-1) x 10 min(-1) IV) displayed no effect on hemodynamics or on coronary sinus or peripheral NE and HA concentrations. These data suggest the potential for therapeutic effect with the latter mechanism in the absence of peripheral cardiac neurotransmitter release or obvious changes in cardiovascular function. PMID- 19129737 TI - Eplerenone decreases inflammatory foci in spontaneously hypertensive rat hearts with minimal effects on blood pressure. AB - The blood pressure (BP)-lowering effects of mineralocorticoid receptor (MR) antagonists in salt-sensitive rat models of hypertension are well understood. However, studies in salt-independent models have yielded mixed results, and therefore, we measured the hemodynamic effects of MR blockade in spontaneously hypertensive rats. We treated spontaneously hypertensive rats for 8 weeks with 30 300 mg.kg.d eplerenone or 20 mg.kg.d losartan and monitored BP using radiotelemetry and performed histopathological analyses of the hearts. Eplerenone, in contrast to losartan, caused only a small reduction in systolic BP at the highest dose tested. Both reduced left ventricular wall thickness, although eplerenone was less effective than losartan. Only losartan decreased heart weight. We observed foci of cardiomyopathy characterized by combinations of infiltrating monocytes, necrotic myocytes, and interstitial fibrosis in hearts of control animals. The number of foci seemed to be decreased in hearts of losartan- and eplerenone-treated animals. In a second study, using quantitative histomorphometry, the number of foci was significantly reduced by 20 mg.kg.d losartan (by 68%) or by 300 mg.kg.d eplerenone (by 50%). Our data support the hypothesis that a direct BP-independent effect on the progression of cardiomyopathy in the heart may be one basis for the cardiac protection afforded by MR antagonism. PMID- 19129738 TI - Excessive hypertension and end-organ damage in a transgenic mouse line carrying the rat angiotensinogen gene. AB - The renin-angiotensin system plays an important role in the etiology of cardiovascular diseases. Three transgenic mouse lines overexpressing rat angiotensinogen (rAOGEN) were generated. The aim of our study was to characterize the originally undescribed second transgenic line TGM(rAOGEN)102. The transgene tissue distribution and expression of brain natriuretic peptide and collagen type III were investigated by ribonuclease protection assay. Catheter measurements of blood pressure and cardiac function were performed in anesthetized mice. End organ fibrosis was further assessed by van Gieson staining. In line TGM(rAOGEN)102, the rAOGEN transgene was mainly expressed in liver and brain but could also be detected in hearts, kidneys, and lungs. Transgenic mice developed excessive chronic hypertension compared with their wild-type littermates. The rise of blood pressure was paralleled by cardiac hypertrophy, impaired cardiac function, and increased expression of brain natriuretic peptide. Pronounced fibrosis was detected in the hearts, lungs, and kidneys of transgenic mice. Our data indicate that overexpression of rAOGEN in mice leads to excessive hypertension, cardiac hypertrophy, impaired heart function, and pronounced fibrosis. Thus, this line TGM(rAOGEN)102 provides a new model to study hypertension-mediated end-organ damage and to evaluate new antihypertensive or cardioprotective drugs. PMID- 19129739 TI - Does endothelium-derived hyperpolarizing factor play a role in endothelium dependent component of electrical field stimulation-induced vasorelaxation of rat mesenteric arterial rings? AB - Electrical field stimulation (EFS)-induced, nonadrenergic, noncholinergic vasodilation was investigated in rat mesenteric arterial rings. Tetrodotoxin (10( 6) M), capsaicin (10(-5) M), or L-NAME (10(-4) M) failed to change the EFS induced relaxations, whereas they were increased with indomethacin (10(-5) M). Removal of the endothelium caused approximately 20% reduction in the maximum response, whereas precontraction with 40 mM KCI abolished the relaxations at all frequencies. Iberiotoxin (3 x 10(-7) M) attenuated the relaxations in endothelium intact tissues but blocked completely those in endothelium-denuded arteries. Combination of TRAM-34 (10(-5) M) with apamin (5 x 10(-7) M) and single administrations of NiCI2 (5 x 10(-4) M), ruthenium red (3 x 10(-5) M), and 18[alpha]-glycyrrhetinic acid (10(-4) M) significantly reduced the responses only in endothelium-intact tissues. These data indicate that in rat mesenteric arteries, EFS leads to vasodilation through both endothelium-dependent and endothelium-independent mechanisms. The major component of the relaxation is endothelium independent and seems to occur via BK(Ca) channels, whereas endothelium-dependent component is likely to be mediated by endothelium-derived hyperpolarizing factor rather than nitric oxide, prostacyclin, or a neural substance. We propose that Ca2+ entry into endothelial cells via nonspecific cation channels in response to EFS induces hyperpolarization by activating endothelial IK(Ca) and SK(Ca) channels, which is spread to the smooth muscle via gap junctions to produce relaxation. PMID- 19129740 TI - Effects of acetylsalicylic acid on increase of fibrin network porosity and the consequent upregulation of fibrinolysis. AB - Our earlier study in vivo showed that a lower dose of acetylsalicylic acid (ASA) brought greater enhancement in fibrin gel permeability (Ks) than a higher dose. To assess whether this finding related to modifications of fibrinogen clotting property by ASA, purified fibrinogen was incubated with ASA and/or salicylic acid (SA). The fibrinogen product was examined. Fibrinogen "clotting time" was not affected. Shortening of fibrin clot "lysis time" paralleled the increase of fibrin network porosity demonstrated by measurements of liquid permeability (Ks), fibrin fiber thickness, and 3-dimensional microscopic image, in a low ASA concentration-dependent way. Ks levels were not altered by SA alone but significantly decreased in samples treated by both where the concentrations were low for ASA and high for SA. In conclusion, ASA at the concentrations used did not influence the rate of fibrinogen gelation by thrombin. However, assembly of fibrin monomers was most probably altered, leading to enhancement of fibrin fiber thickness. A looser network was constructed by the thicker fibrin fibers, which benefits fibrinolysis. According to the known mechanism that SA interferes with ASA in preventing acetylation of platelet's proteins, an explanation for the low ASA concentration-dependent effects on fibrin network structure may be that fewer molecules of SA-the hydrolytic product of ASA-are generated from lower doses of ASA, which block acetylation of fibrinogen to a smaller extent and thus more significantly impair fibrin formation. PMID- 19129741 TI - Gene transfer to mouse heart and skeletal muscles using a minicircle expressing human vascular endothelial growth factor. AB - BACKGROUND: : Gene transfer to heart muscle is a promising modality to treat ischemic heart disease. However, current vectors are inefficient and need to be improved. A novel vector system that shows great promise is the minicircle (MC) vector being smaller than conventional plasmid vectors and devoid of bacterial sequences. AIMS: : To study gene transfer of MC DNA, expressing the human vascular endothelial growth factor (hVEGF), to mouse heart and skeletal muscles and to compare it with one of the efficient plasmids used in cardiovascular trials, the phVEGF165 containing the same expression cassette as the MC. RESULTS: : The MC and the phVEGF165 plasmid show similar expression patterns both in vitro and in mouse heart and skeletal muscle studies in vivo on molar basis (equal expression in heart 24 hours, 0.9 fold lower expression from MC in heart and 1.9 fold higher in skeletal muscle at 7 days), whereas on weight basis the MC construct was more efficient in skeletal muscle (5.6 fold higher expression, P < 0.05), and at least as efficient in heart (1.6 fold higher expression). CONCLUSIONS: : The gene expression is similar in the 2 vector systems, so the smaller size and the fact that the MC construct is devoid of bacterial sequences and antibiotics resistance gene make the MC vector an attractive alternative for nonviral gene therapy. PMID- 19129742 TI - Effects of ivabradine and metoprolol on cardiac angiogenesis and endothelial dysfunction in rats with heart failure. AB - Myocardial infarction (MI)-induced remodeling is associated with disturbed myocardial perfusion through vascular changes, such as reduced capillary density and endothelial dysfunction. Heart rate reduction (HRR) initiated immediately after MI stimulates angiogenesis and attenuates left ventricular dysfunction. We aimed to investigate the effects of long-term HRR on cardiac angiogenesis and endothelial function in a rat model of post-MI heart failure. Rats received early or late ivabradine or metoprolol for 12 or 9 weeks, respectively, and compared with untreated MI and sham animals 12 weeks after MI. Heart rate was measured in the conscious rat. MI resulted in an increased heart weight to body weight ratio, a decline in capillary density and a marked reduction in acetylcholine-induced relaxation. Early and late HRR by either ivabradine or metoprolol significantly increased capillary to myocyte ratio. Moreover, this ratio was significantly correlated to heart rate (r = -0.324 and P = 0.036). Neither early nor late chronic HRR prevented endothelial dysfunction, except a moderate improvement in late MI ivabradine group. In MI rats, HRR either by ivabradine or metoprolol treatment increases cardiac angiogenesis. Late HRR strategy was comparable to early HRR, suggesting that the beneficial effects are independent of the time of onset of therapy after MI. PMID- 19129743 TI - Experimental hyperlipidemia causes an increase in the electrocardiographic changes associated with amiodarone. AB - To assess the influence of hyperlipidemia (HL) on amiodarone (AM) effect in the heart, rats were pretreated with either 1 g/kg poloxamer 407 (to induce HL) or saline intraperitoneally. At approximately 36 hours afterward, rats were given AM HCl (25, 50, and 100 mg.kg.d) or saline intravenously through implanted venous cannulas for 5 days. Under anesthesia, electrocardiogram (ECG) was recorded using subcutaneous electrodes and blood samples were withdrawn at baseline and 12 hours after the first, middle, and last doses. At the end of the study, heart tissues were collected. Specimens were analyzed for AM and desethylamiodarone. HL by itself did not alter the ECG. Compared with baseline, the end of study prolongation of QTc and PR intervals were significantly (P < 0.05) higher in all AM-treated HL rats. AM plasma and heart concentrations in HL rats after the last dose were significantly (P < 0.05) higher than in normolipidemic rats. Similar to AM, in HL rats, plasma desethylamiodarone after the last dose was significantly higher than in normolipidemic rats. The cholesterol to triglyceride plasma ratio was linearly related to QT interval and plasma and heart AM concentrations. HL increased the ECG effects of AM by increasing heart concentrations. PMID- 19129744 TI - Racial/ethnic differences in the birth prevalence of spina bifida - United States, 1995-2005. AB - In 1992, the U.S. Public Health Service recommended that all women of childbearing age consume 400 microg of folic acid daily to help prevent pregnancies affected by neural tube defects (NTDs) such as spina bifida. Subsequently, the Food and Drug Administration mandated adding folic acid to all enriched cereal grain products by January 1998. During October 1998--December 1999, the birth prevalence of spina bifida in the United States decreased 22.9% compared with 1995--1996; however, by 2003--2004, no further decrease had been observed. Notably, the prevalence of NTD-affected pregnancies remained higher among Hispanic women than among women in other racial/ethnic populations. To update previously reported data and assess racial/ethnic differences, CDC analyzed birth certificate data for four periods during 1995--2005. This report summarizes the results of that analysis, which indicated that from the early postfortification period, 1999--2000, to the most recent period of analysis, 2003 -2005, the prevalence of spina bifida declined 6.9%, from 2.04 to 1.90 per 10,000 live births (prevalence ratio [PR] = 0.93; 95% confidence interval [CI] = 0.87- 1.00). Among infants with non-Hispanic black mothers, prevalence fell 19.8%, from 2.17 to 1.74 per 10,000 live births (PR = 0.80; CI = 0.67--0.96), while prevalence among infants with non-Hispanic white and Hispanic mothers remained nearly constant. Additional public health efforts targeting women with known risk factors (e.g., obesity and certain genetic factors) likely are needed to further reduce the prevalence of spina bifida in the United States. PMID- 19129745 TI - Investigation of patients treated by an HIV-infected cardiothoracic surgeon- Israel, 2007. AB - Transmission of human immunodeficiency virus (HIV) from an infected health-care worker to patients is rare, with the greatest potential for occurrence during exposure-prone, invasive surgical procedures in which the blood of the health care worker might come into contact with patients' blood or mucous membranes. When a surgeon is discovered to have HIV infection, a decision must be made about notification of patients, but only limited data are available to guide decision making. Such notifications generally are decided upon on a case-by-case basis, taking into account such factors as the nature of the procedures performed, the infection-control knowledge and practices of the infected surgeon, the presumed likelihood of transmission, and available resources. This report describes the case of a cardiothoracic surgeon in Israel specializing in open-heart procedures (coronary artery bypass grafting and valve surgery) who was found to be HIV positive in January 2007 during evaluation for fever of recent onset. The duration of infection was unknown. A lookback investigation of patients operated on by the infected surgeon during the preceding 10 years was conducted under the auspices of the Israel Ministry of Health to determine whether any surgeon-to patient HIV transmission had occurred. Of 1,669 patients identified, 545 (33%) underwent serologic testing for HIV antibody. All results were negative. A Ministry-appointed panel of experts delineated conditions under which the surgeon could resume work. The results of this investigation add to previously published data indicating a low risk for provider-to-patient HIV transmission. PMID- 19129746 TI - Changes in tobacco use among youths aged 13-15 years - Panama, 2002 and 2008. AB - Tobacco use is the single most preventable cause of death in the world today, and the majority of smokers begin using tobacco products before age 18 years. However, before the late 1990s, few countries had reliable data on youth tobacco use. In 1999, the World Health Organization (WHO), CDC, and the Canadian Public Health Association developed the Global Youth Tobacco Survey (GYTS) to help countries monitor youth tobacco use. At the same time, WHO initiated the Framework Convention on Tobacco Control (WHO FCTC), the first international public health treaty on tobacco control. Panama ratified WHO FCTC in 2004 and enacted two key antitobacco regulations in 2005 and 2008. To evaluate progress toward attaining tobacco control goals in Panama, Panama's Ministry of Health, CDC, and WHO compared results from GYTS surveys conducted in Panama in 2002 and 2008. This report summarizes the results of that comparison, which revealed substantial decreases from 2002 to 2008 in youth current cigarette smoking (13.2% versus 4.3%), current use of tobacco products other than cigarettes (9.8% versus 5.8%), and likely initiation of smoking by never smokers (13.8% versus 10.0%). In addition, factors influencing tobacco use showed substantial decreases, including 1) exposure to secondhand smoke (SHS) at home and in public places, 2) best friends smoking, 3) protobacco advertising in newspapers and magazines, and 4) having an object with a tobacco company logo on it. These results suggest that comprehensive regulations in Panama helped reduce tobacco use among adolescents and further gains are possible. PMID- 19129747 TI - Pro32Thr polymorphism of inosine triphosphate pyrophosphatase gene predicts efficacy of low-dose azathioprine for patients with systemic lupus erythematosus. AB - We evaluated the relationship between the efficacy of low-dose azathioprine (AZA) therapy and the inosine triphosphate pyrophosphatase (ITPA) 94C>A (Pro32Thr) polymorphism in patients with systemic lupus erythematosus (SLE). We performed a multiple regression analysis to assess the influence of various factors on the reduction in SLE disease activity index (SLEDAI) scores. The ITPA 94C>A polymorphism had the highest correlation with the change in SLEDAI score (r = 0.354, P = 0.006). PMID- 19129748 TI - Dapagliflozin, a novel SGLT2 inhibitor, induces dose-dependent glucosuria in healthy subjects. AB - Dapagliflozin selectively inhibits renal glucose reabsorption by inhibiting sodium-glucose cotransporter-2 (SGLT2). It was developed as an insulin independent treatment approach for type 2 diabetes mellitus (T2DM). The safety, tolerability, pharmacokinetics, and pharmacodynamics of the drug were evaluated in single-ascending-dose (SAD; 2.5-500 mg) and multiple-ascending-dose (MAD; 2.5 100 mg daily for 14 days) studies in healthy subjects. Dapagliflozin exhibited dose-proportional plasma concentrations with a half-life of approximately 17 h. The amount of glucosuria was also dose-dependent. Cumulative amounts of glucose excreted on day 1, relating to doses from 2.5-100 mg (MAD), ranged from 18 to 62 g; day 14 values were comparable to day 1 values, with no apparent changes in glycemic parameters. Doses of approximately 20-50 mg provided close-to-maximal SGLT2 inhibition for at least 24 h. Dapagliflozin demonstrates pharmacokinetic (PK) characteristics and dose-dependent glucosuria that are sustained over 24 h, which indicates that it is suitable for administration in once-daily doses and suggests that further investigation of its efficacy in T2DM patients is warranted. PMID- 19129750 TI - Monitoring for hepatotoxicity: what is the predictive value of liver "function" tests? AB - Drug-induced liver injury (DILI) is a major reason drugs fail during development or are withdrawn from the market. The ability to predict, detect, and avoid DILI through appropriate patient selection and effective monitoring has proved to be an elusive goal. Many approved drugs have labeling recommendations for serum enzyme monitoring intended to detect and prevent hepatotoxicity, but such monitoring is often seen as inconvenient, uncomfortable, costly, and inefficient by both patients and doctors, and thus monitoring recommendations are poorly followed, if at all. This review considers whether monitoring works to prevent DILI, whether monitoring recommendations are derived from data or opinions, and whether any better alternatives exist. PMID- 19129749 TI - Dapagliflozin, a novel, selective SGLT2 inhibitor, improved glycemic control over 2 weeks in patients with type 2 diabetes mellitus. AB - Dapagliflozin, administered to patients in once-daily oral doses, is a sodium glucose cotransporter 2 (SGLT2) inhibitor that blocks the reabsorption of glucose from urine into the blood. This 14-day study randomized patients with type 2 diabetes mellitus (T2DM) to four treatment groups receiving daily oral doses of 5 , 25-, or 100-mg doses of dapagliflozin or placebo, in order to evaluate glucosuria and glycemic parameters. Significant reductions in fasting serum glucose (FSG) were observed on day 2 with 100 mg dapagliflozin (-9.3%, P < 0.001), and dose-dependent reductions were observed on day 13 with the 5-mg ( 11.7%; P < 0.05), 25-mg (-13.3%; P < 0.05), and 100-mg (-21.8%; P < 0.0001) doses as compared with placebo. Significant improvements in oral glucose tolerance test (OGTT) were observed with all doses on days 2 and 13 (P < 0.001 as compared with placebo). On day 14, urine glucose values were 36.6, 70.1, and 69.9 g/day for the 5-, 25-, and 100-mg doses (as compared with no change for placebo), which were slightly lower than those on day 1. This was attributed to the decrease in filtered glucose load following improved glycemic control. Dapagliflozin produced dose-dependent increases in glucosuria and clinically meaningful changes in glycemic parameters in T2DM patients. PMID- 19129751 TI - Assessment of MAO-B occupancy in the brain with PET and [11C]-L-deprenyl-D2: a dose-finding study with a novel MAO-B inhibitor, EVT 301. AB - Inhibition of monoamine oxidase type B (MAO-B) activity in the brain is a putative strategy for the treatment of Alzheimer's disease (AD). We performed a dose-selection and validation study of a novel, reversible MAO-B inhibitor, EVT 301. Sixteen healthy volunteers received selegiline (10 mg) or EVT 301 (25, 75, or 150 mg) daily for 7-8 days, and four subjects with AD received 75 mg of EVT 301. MAO-B occupancy in the brain was assessed using positron emission tomography (PET) with [11C]-L-deprenyl-D2. EVT 301 was found to dose-dependently occupy MAO B in the human brain, with occupancy ranging from 58-78% at a dose of 25 mg to 73 90% at a dose of 150 mg. The corresponding occupancy after selegiline was 77-92%. Determination of MAO-B inhibition in blood platelets underestimated the actual brain occupancy achieved with EVT 301. A daily EVT 301 dose of 75 or 150 mg appears suitable for clinical efficacy studies in patients with AD. PMID- 19129752 TI - Probiotic E. coli treatment mediates antimicrobial human beta-defensin synthesis and fecal excretion in humans. AB - Inducible epithelial human beta-defensins (hBD) play an important role in intestinal barrier function. In vitro studies showed that clinically effective probiotics induce antimicrobial hBD-2. Here, we aimed to assess the in vivo effect in healthy volunteers and also addressed how defensins affect probiotic survival. Symbioflor 2 containing one strain of several viable genotypes of Escherichia coli was administered to 23 healthy individuals. After 3 weeks, fecal hBD-2 peptide was increased in 78% (mean 3.7-fold; P<0.0001). Interestingly, the fecal hBD-2 peptide was still elevated 9 weeks after treatment (P=0.008). In vitro studies revealed that this effect was mediated by only one out of three tested E. coli genotypes and comparable to probiotic E. coli Nissle 1917 (10- to 15-fold). Functional assays showed that all tested bacteria were similarly killed by defensins allowing to speculate about a suicidal character of this effect. Defensin induction seems to be a common and important mechanism of probiotic treatment. PMID- 19129753 TI - Crossing barriers: infections of the lung and the gut. AB - Although known as respiratory pathogens, severe acute respiratory syndrome (SARS) and its sister coronaviruses frequently cause enteric symptoms. In addition, other classically non-enteric viruses (such as HIV and influenza) may also have enteric effects that are crucial in their pathogeneses. These effects can be due to direct infection of the gut mucosa, but can also be because of decreased antibacterial defenses, increased mucosal permeability, bacterial translocation, and systemic leak of endotoxin. PMID- 19129754 TI - Steady-state migrating intestinal dendritic cells induce potent inflammatory responses in naive CD4+ T cells. AB - Steady-state dendritic cells (DCs) migrating in the lymph from the intestine induce tolerance to harmless intestinal antigens, preventing inflammatory responses. To determine if such DCs are inherently tolerogenic we collected intestinal lymph DCs (L-DCs) by cannulation of the thoracic duct of rats after mesenteric lymphadenectomy, and examined their capacity to activate naive CD4+ lymphocytes in an allogeneic mixed leucocyte reaction. L-DCs stimulated strong proliferative responses, induced secretion of inflammatory cytokines including interferon-gamma, and induced FoxP3-positive lymphocytes to divide. To determine if the activated CD4+ T cells had been tolerized, they were rested and restimulated with irradiated splenocytes. The restimulated CD4+ T cells again proliferated and secreted inflammatory cytokines. These data demonstrate that the DCs, which migrate from the intestine in the steady state, are paradoxically able to induce strong inflammatory responses from naive T cells, despite their role in the maintenance of oral tolerance. PMID- 19129755 TI - Suppression of airway inflammation by a natural acute infection of the intestinal epithelium. AB - Although chronic intestinal helminth infections may suppress allergen-induced airway pathology by inducing a combination of modified T-helper (Th) 2 and immunosuppressive cytokines, a similar capacity of natural acute intestinal infections has remained untested, despite their global prevalence. Here, we show that allergic airway phenotypes including eosinophilia, eotaxin mRNA, and Th2 cytokines are significantly suppressed in animals that were infected by and that have cleared the intestinal parasite Eimeria vermiformis. Unlike in helminth infected animals, regulation requires temporal coincidence of infection with sensitization; depends on interferon-gamma; and is not associated with an enhanced antigen-specific immunoglobulin G1 response. Moreover, regulation was effective following allergen sensitization in different anatomical sites, and in young and adult mice. These data highlight a transient anatomical dissemination of "functional immunologic dominance" following infection of the gut mucosa. They strongly support the hypothesis that airway allergies are naturally suppressed by both acute and chronic mucosal pathogens, but by different mechanisms. PMID- 19129756 TI - A genital tract peptide epitope vaccine targeting TLR-2 efficiently induces local and systemic CD8+ T cells and protects against herpes simplex virus type 2 challenge. AB - The next generation of needle-free mucosal vaccines is being rationally designed according to rules that govern the way in which the epitopes are recognized by and stimulate the genital mucosal immune system. We hypothesized that synthetic peptide epitopes extended with an agonist of Toll-like receptor 2 (TLR-2), that are abundantly expressed by dendritic and epithelial cells of the vaginal mucosa, would lead to induction of protective immunity against genital herpes. To test this hypothesis, we intravaginally (IVAG) immunized wild-type B6, TLR-2 (TLR2(-/ )) or myeloid differentiation factor 88 deficient (MyD88(-/-)) mice with a herpes simplex virus type 2 (HSV-2) CD8+ T-cell peptide epitope extended by a palmitic acid moiety (a TLR-2 agonist). IVAG delivery of the lipopeptide generated HSV-2 specific memory CD8+ cytotoxic T cells both locally in the genital tract draining lymph nodes and systemically in the spleen. Moreover, lipopeptide-immunized TLR2( /-) and MyD88(-/-) mice developed significantly less HSV-specific CD8+ T-cell response, earlier death, faster disease progression, and higher vaginal HSV-2 titers compared to lipopeptide-immunized wild-type B6 mice. IVAG immunization with self-adjuvanting lipid-tailed peptides appears to be a novel mucosal vaccine approach, which has attractive practical and immunological features. PMID- 19129757 TI - The roles of CCR6 in migration of Th17 cells and regulation of effector T-cell balance in the gut. AB - Migration and trafficking receptors of Th17 cells to mucosal tissues have been unclear. We report that Th17 cells preferentially migrate to the intestine and associated lymphoid tissues, and CCR6 is the homing receptor important for Th17 cell migration to certain tissue microenvironments of the intestine such as Peyer's patches and other sites where its ligand CCL20 is expressed. We found the cytokine transforming growth factor-beta1 is required for CCR6 expression whereas IL-2 suppresses it. CCR6-deficient Th17 cells aberrantly migrate to different compartments of the intestine. Surprisingly, administration of CCR6-deficient Th17 cells into severe combined immunodeficiency (SCID) mice led to excessive intestinal inflammation with increased Th1 but decreased Th17 cells and FoxP3(+) T cells. In addition, CCR6 deficiency led to aberrantly widespread effector T cells in the inflamed intestine of the SCID mice. We conclude that CCR6 regulates Th17 cell migration to the gut and effector T-cell balance/distribution in inflamed intestine. PMID- 19129759 TI - At the crossroads: mucosal immunology of the larynx. AB - The larynx sits at the crossroads between gastrointestinal and respiratory tracts. Besides its intrinsic importance in breathing, swallowing and voice production, the larynx is also exposed to unique immunological challenges. Given the propensity of chronic inflammatory conditions such as chronic laryngitis, which affects up to 20% of Western populations, it is surprising that our understanding of the immunology of this organ remains relatively limited. Recent work on the immunological architecture of the laryngeal mucosa, and its changes that result from external challenges and inflammatory conditions, provided valuable insight into the fascinating immunology of this organ. The lessons learnt from these investigations may go beyond devising improved therapy for chronic laryngeal inflammation. Establishing whether and how the laryngeal mucosa may be involved in the modulation of wider mucosal responses may provide novel routes to the treatment of inflammatory diseases of the respiratory and alimentary tracts such as asthma and inflammatory bowel disease. PMID- 19129760 TI - TRPCs, GPCRs and the Bayliss effect. PMID- 19129761 TI - One for all? A viral protein supplants the mRNA cap-binding complex. PMID- 19129762 TI - Runx1 is required for the endothelial to haematopoietic cell transition but not thereafter. AB - Haematopoietic stem cells (HSCs) are the founder cells of the adult haematopoietic system, and thus knowledge of the molecular program directing their generation during development is important for regenerative haematopoietic strategies. Runx1 is a pivotal transcription factor required for HSC generation in the vascular regions of the mouse conceptus-the aorta, vitelline and umbilical arteries, yolk sac and placenta. It is thought that HSCs emerge from vascular endothelial cells through the formation of intra-arterial clusters and that Runx1 functions during the transition from 'haemogenic endothelium' to HSCs. Here we show by conditional deletion that Runx1 activity in vascular-endothelial-cadherin positive endothelial cells is indeed essential for intra-arterial cluster, haematopoietic progenitor and HSC formation in mice. In contrast, Runx1 is not required in cells expressing Vav1, one of the first pan-haematopoietic genes expressed in HSCs. Collectively these data show that Runx1 function is essential in endothelial cells for haematopoietic progenitor and HSC formation from the vasculature, but its requirement ends once or before Vav is expressed. PMID- 19129758 TI - Pulmonary fibrosis: pathogenesis, etiology and regulation. AB - Pulmonary fibrosis and architectural remodeling of tissues can severely disrupt lung function, often with fatal consequences. The etiology of pulmonary fibrotic diseases is varied, with an array of triggers including allergens, chemicals, radiation and environmental particles. However, the cause of one of the most common pulmonary fibrotic conditions, idiopathic pulmonary fibrosis (IPF), is still unclear. This review examines common mechanisms of pulmonary wound-healing responses following lung injury, and highlights the pathogenesis of some of the most widespread pulmonary fibrotic diseases. A three phase model of wound repair is reviewed that includes; (1) injury; (2) inflammation; and (3) repair. In most pulmonary fibrotic conditions dysregulation at one or more of these phases has been reported. Chronic inflammation can lead to an imbalance in the production of chemokines, cytokines, growth factors, and disrupt cellular recruitment. These changes coupled with excessive pro-fibrotic IL-13 and/or TGFbeta1 production can turn a well-controlled healing response into a pathogenic fibrotic response. Endogenous regulatory mechanisms are discussed including novel areas of therapeutic intervention. Restoring homeostasis to these dysregulated healing responses, or simply neutralizing the key pro-fibrotic mediators may prevent or slow the progression of pulmonary fibrosis. PMID- 19129764 TI - New Year's resolutions. PMID- 19129765 TI - Challenges anticipated in removal of stem cell restrictions. PMID- 19129763 TI - Intersubunit coordination in a homomeric ring ATPase. AB - Homomeric ring ATPases perform many vital and varied tasks in the cell, ranging from chromosome segregation to protein degradation. Here we report the direct observation of the intersubunit coordination and step size of such a ring ATPase, the double-stranded-DNA packaging motor in the bacteriophage phi29. Using high resolution optical tweezers, we find that packaging occurs in increments of 10 base pairs (bp). Statistical analysis of the preceding dwell times reveals that multiple ATPs bind during each dwell, and application of high force reveals that these 10-bp increments are composed of four 2.5-bp steps. These results indicate that the hydrolysis cycles of the individual subunits are highly coordinated by means of a mechanism novel for ring ATPases. Furthermore, a step size that is a non-integer number of base pairs demands new models for motor-DNA interactions. PMID- 19129766 TI - A pipe dream becomes reality. PMID- 19129767 TI - Religious leaders weigh in on current and future embryo research. PMID- 19129768 TI - Report details scientific self-censoring. PMID- 19129769 TI - HPV researchers set sights on preventing skin cancers. PMID- 19129771 TI - Straight talk with... Mervyn Turner. Interview by Prashant Nair. PMID- 19129772 TI - Mended armor. PMID- 19129774 TI - Moving neurons back into place. PMID- 19129775 TI - Mesenchymal stem cells combat sepsis. PMID- 19129776 TI - Gamma-secretase inhibitors: Notch so bad. PMID- 19129777 TI - Fixing a failed vaccine. PMID- 19129778 TI - Cancer stem cells are everywhere. PMID- 19129779 TI - Rescuing a failing heart: putting on the squeeze. PMID- 19129780 TI - Rescuing a failing heart: think globally, treat locally. PMID- 19129784 TI - Food for thought. PMID- 19129785 TI - The ketogenic diet, four score and seven years later. AB - This Practice Point commentary discusses the findings of Neal et al.'s randomized controlled trial of the ketogenic diet in children with refractory epilepsies. The authors showed that the ketogenic diet was superior to continuation of medical treatment in reducing seizure frequency in this patient population. On the basis of these and other results I argue three points, the first being supported by the literature and the others being personal opinion. First, the ketogenic diet should be considered for the treatment of children with drug resistant epilepsy and not only as a last resort. Second, the allocation of resources to support professionals in administering the ketogenic diet is justifiable. Third, we must develop alternative strategies to randomized controlled trials if we wish to obtain timely information on effective treatment strategies for specific pediatric epilepsy syndromes. PMID- 19129786 TI - Does prolonged skin temperature measurement improve the diagnosis of complex regional pain syndrome? AB - This Practice Point provides commentary on a prospective study by Krumova et al. that explored the diagnostic utility of long-term temperature measurements in patients with upper extremity complex regional pain syndrome (CRPS). The objectives of the group were twofold: to evaluate vascular abnormalities in CRPS by comparing real-time thermal variations experienced in everyday circumstances over prolonged time frames in affected and nonaffected hands, and to develop a practical approach for differentiating CRPS from other painful conditions. Measurement of skin temperature dynamics differentiated between CRPS and arm pain secondary to other etiologies with a sensitivity of 73% and a specificity of 67%. Although the technique Krumova and colleagues used is more practical than those previously described, it is still too onerous for patients and physicians to routinely employ. We anticipate that improved identification of pain mechanisms will translate into better treatment outcomes, but this hypothesis remains to be tested. PMID- 19129787 TI - Folic acid supplementation for women with epilepsy who might become pregnant. AB - Low folate levels are associated with an increased risk of spontaneous abortion and congenital malformations. Up to half of all pregnancies are unplanned, so official guidelines in a number of countries, including the US, recommend folic acid supplementation for all women who might become pregnant, before they actually start planning to have children. Children born to women who are taking antiepileptic medications, many of which antagonize folate action, are at an increased risk of birth defects. This article highlights a study by Pittschieler et al. that has suggested that supplementation with folic acid might be particularly important in women with epilepsy who are taking antiepileptic medications--especially valproate. However, the sample size in the study was too small to determine the optimum dose or to detect whether folic acid protects against birth defects. Nonetheless, it is clear that all women of reproductive age, especially those taking certain antiepileptic medications, should take folic acid supplements. PMID- 19129788 TI - Mirror neurons and their clinical relevance. AB - One of the most exciting events in neurosciences over the past few years has been the discovery of a mechanism that unifies action perception and action execution. The essence of this 'mirror' mechanism is as follows: whenever individuals observe an action being done by someone else, a set of neurons that code for that action is activated in the observers' motor system. Since the observers are aware of the outcome of their motor acts, they also understand what the other individual is doing without the need for intermediate cognitive mediation. In this Review, after discussing the most pertinent data concerning the mirror mechanism, we examine the clinical relevance of this mechanism. We first discuss the relationship between mirror mechanism impairment and some core symptoms of autism. We then outline the theoretical principles of neurorehabilitation strategies based on the mirror mechanism. We conclude by examining the relationship between the mirror mechanism and some features of the environmental dependency syndromes. PMID- 19129790 TI - A case of maternal herpes simplex virus encephalitis during late pregnancy. AB - BACKGROUND: A pregnant 25-year-old woman at 32 weeks' gestation was admitted to an emergency unit after her husband had found her drowsy and with her tongue bitten. The day before admission, the patient had developed a fever of 39 degrees C, was suffering from headaches, was nauseated and had vomited. On admission, she had anterograde and retrograde amnesia, but no somatic neurological deficits were detected. INVESTIGATIONS: Routine laboratory testing, lumbar puncture, cerebrospinal fluid analysis, routine bacteriology, brain MRI, and polymerase chain reaction testing for neurotropic viruses including herpes simplex virus types 1 and 2. DIAGNOSIS: Maternal herpes simplex virus type 1 encephalitis. MANAGEMENT: Antiviral and anticonvulsive therapy, supportive treatment, and cesarean section. PMID- 19129789 TI - Infant pain management: a developmental neurobiological approach. AB - Infant pain is a clinical reality. Effective pain management in infants requires a specialist approach--analgesic protocols that have been designed for older children cannot simply be scaled down for CNS pain pathways and analgesic targets that are in a state of developmental transition. Here, we discuss the particular challenges that are presented by an immature CNS for the detection and treatment of pain. We show how the application of neurophysiological and neuropharmacological approaches can help to overcome the problems inherent in measuring and treating pain in infants, and how research data in these areas can be used to devise age-appropriate methods of assessing pain as well as strategies for pain relief. The evidence that untreated pain in infancy results in long-term adverse consequences is presented, thereby emphasizing the need for a longer term view of infant pain management. PMID- 19129792 TI - Autonomic function after spinal cord lesion: animal guidelines and international standards. PMID- 19129791 TI - Making a commitment: cell lineage allocation and axis patterning in the early mouse embryo. AB - Genetic studies have identified the key signalling pathways and developmentally regulated transcription factors that govern cell lineage allocation and axis patterning in the early mammalian embryo. Recent advances have uncovered details of the molecular circuits that tightly control cell growth and differentiation in the mammalian embryo from the blastocyst stage, through the establishment of initial anterior-posterior polarity, to gastrulation, when the germ cells are set aside and the three primary germ layers are specified. Relevant studies in lower vertebrates indicate the conservation and divergence of regulatory mechanisms for cell lineage allocation and axis patterning. PMID- 19129793 TI - Effect of bisphosphonates on vascular calcification and bone metabolism in experimental renal failure. AB - Although it is known that bisphosphonates prevent medial vascular calcification in vivo, their mechanism of action remains unknown and, in particular, whether they act directly on the blood vessels or indirectly through inhibition of bone resorption. To determine this, we studied the effects of two bisphosphonates on calcification of rat aortas in vitro and on in vivo aortic calcification and bone metabolism in rats with renal failure. We produced vascular calcification in rats with adenine-induced renal failure fed a high-phosphate diet. Daily treatment with either etidronate or pamidronate prevented aortic calcification, with the latter being 100-fold more potent. Both aortic calcification and bone formation were reduced in parallel; however, bone resorption was not significantly affected. In all uremic rats, aortic calcium content correlated with bone formation but not with bone resorption. Bisphosphonates also inhibited calcification of rat aortas in culture and arrested further calcification of precalcified vessels but did not reverse their calcification. Expression of osteogenic factors or calcification inhibitors was not altered by etidronate in vitro. Hence, these studies show that bisphosphonates can directly inhibit uremic vascular calcification independent of bone resorption. The correlation between inhibition of aortic calcification and bone mineralization is consistent with a common mechanism such as the prevention of hydroxyapatite formation and suggests that bisphosphonates may not be able to prevent vascular calcification without inhibiting bone formation in uremic rats. PMID- 19129794 TI - Phosphate and the parathyroid. AB - The phosphate (Pi) retention in patients with chronic kidney disease leads to secondary hyperparathyroidism (2HPT). 2HPT is the physiological response of the parathyroid not only to Pi retention but also to decreased synthesis of 1,25(OH)(2) vitamin D, and the attendant hypocalcemia. 2HPT is characterized by increased PTH synthesis, secretion, and parathyroid cell proliferation. Extracellular fluid (ECF) Ca(2+) is recognized by the parathyroid calcium receptor and a small decrease in the ECF Ca(2+) results in relaxation of the calcium receptor and allows the unrestrained secretion and synthesis of PTH and in the longer term, parathyroid cell proliferation. Both 1,25(OH)(2) vitamin D and fibroblast growth factor 23 inhibit PTH gene expression and secretion. Secondary hyperparathyroidism can initially be controlled by a single therapeutic intervention, such as a Pi-restricted diet, a calcimimetic, or an active vitamin D analog. In this review we discuss the mechanisms whereby Pi regulates the parathyroid. Pi has a direct effect on the parathyroid which requires intact parathyroid tissue architecture. The effect of Pi, as of Ca(2+), on PTH gene expression is post-transcriptional and involves the regulated interaction of parathyroid cytosolic proteins to a defined cis acting sequence in the PTH mRNA. Changes in serum Ca(2+) or Pi regulate the activity of trans acting interacting proteins in the parathyroid, which alters their binding to a defined 26 nucleotide cis acting instability sequence in the PTH mRNA 3'-untranslated region. The trans factors are either stabilizing or destabilizing factors and their regulated binding to the PTH cis acting element determines the PTH mRNA half-life. The responses of the parathyroid to changes in serum Pi are now being revealed but the sensing mechanisms remain a mystery. PMID- 19129796 TI - Cesarean risk after successful external cephalic version: a matched, retrospective analysis. AB - OBJECTIVE: To determine the odds of cesarean, operative vaginal delivery and vaginal birth after cesarean after successful external cephalic version (ECV) compared with singleton pregnancies eligible for a trial of labor. STUDY DESIGN: A matched case-control study was performed using the Memorial Care OBStat Database from 1 January 1998 to 31 July 2006. We identified 197 participants who underwent a successful ECV (study group) and compared them with the next two women presenting for labor management, matched for parity, gestational age, delivery history (previous cesarean delivery) and type of labor (spontaneous or induced). RESULT: There was no significant difference in the adjusted matched odds of cesarean delivery between the study group and control group overall (16.8 vs 11.9%; odds ratio (OR) 1.70; 95% confidence interval (CI) 0.98 to 2.97), even when subanalyzed according to parity. There was also no significant difference in adjusted matched odds of operative vaginal delivery for the study group and control group, 15.9 vs 8.9% (OR 1.06; 95% CI 0.32 to 3.51). Among patients with a prior cesarean, those who underwent successful ECV had a cesarean delivery rate of 11.1% compared with 16.7% in the matched control group (OR 0.59; 95% CI 0.47 to 7.43). CONCLUSION: Cesarean delivery and operative vaginal delivery rates following successful ECV are not increased in our data set compared with matched controls, even in patients with a prior cesarean delivery. This information may be useful when counseling patients who are contemplating an ECV attempt due to non-cephalic presentation at term. PMID- 19129800 TI - Culture clash. PMID- 19129795 TI - Compartmentalization of neutrophils in the kidney and lung following acute ischemic kidney injury. AB - During renal ischemia-reperfusion, local and distant tissue injury is caused by an influx of neutrophils into the affected tissues. Here we measured the kinetics of margination and transmigration of neutrophils in vivo in the kidney and lungs following renal ischemia-reperfusion. After bilateral renal injury, kidney neutrophil content increased threefold at 24 h. The neutrophils were found primarily in the interstitium and to a lesser degree marginated to the vascular endothelium. These interstitial neutrophils had significantly lower levels of intracellular IFN-gamma, IL-4, IL-6, and IL-10 a tendency for decreased amounts of IL-4 and TNF-alpha compared to the marginated neutrophils. Localization of the neutrophils to the kidney interstitium was confirmed by high resolution microscopy and these sites of transmigration were directly associated with areas of increased vascular permeability. Activation of the adenosine 2A receptor significantly decreased both kidney neutrophil transmigration by about half and vascular permeability by about a third. After unilateral renal ischemia reperfusion, the unclipped kidney and lungs did not accumulate interstitial neutrophils or have increased vascular permeability despite a marked increase of neutrophil margination in the lungs. Our findings suggest there is a sequential recruitment and transmigration of neutrophils from the vasculature into the kidney interstitium at the site of tissue injury following renal ischemia reperfusion. PMID- 19129801 TI - Data for the masses. PMID- 19129802 TI - Cuba's biotech boom. PMID- 19129809 TI - Journal club. A crystallographer takes a jaunt into immunology. PMID- 19129814 TI - Biotechs feel the pain. PMID- 19129815 TI - European boost for particle therapy. PMID- 19129823 TI - Ocean research: The lab at the bottom of the sea. PMID- 19129824 TI - Forestry: A green investment. PMID- 19129825 TI - 'Subspecies' and 'race' should not be used as synonyms. PMID- 19129826 TI - Western prosperity is based on resources that are running out. PMID- 19129827 TI - Replacement of animals in research will never be possible. PMID- 19129828 TI - Being human: love: neuroscience reveals all. PMID- 19129834 TI - Organic chemistry: Molecular diversity by design. PMID- 19129836 TI - Immunology: Barrier to electrical storms. PMID- 19129837 TI - Quantum physics: Quantum force turns repulsive. PMID- 19129838 TI - Protein synthesis: Errors rectified in retrospect. PMID- 19129839 TI - Astrophysics: Magnetic bond. PMID- 19129840 TI - Structural biology: Anticancer drug target pictured. PMID- 19129841 TI - Molecular biology: A taste of umami. PMID- 19129842 TI - Energy flux determines magnetic field strength of planets and stars. AB - The magnetic fields of Earth and Jupiter, along with those of rapidly rotating, low-mass stars, are generated by convection-driven dynamos that may operate similarly (the slowly rotating Sun generates its field through a different dynamo mechanism). The field strengths of planets and stars vary over three orders of magnitude, but the critical factor causing that variation has hitherto been unclear. Here we report an extension of a scaling law derived from geodynamo models to rapidly rotating stars that have strong density stratification. The unifying principle in the scaling law is that the energy flux available for generating the magnetic field sets the field strength. Our scaling law fits the observed field strengths of Earth, Jupiter, young contracting stars and rapidly rotating low-mass stars, despite vast differences in the physical conditions of the objects. We predict that the field strengths of rapidly rotating brown dwarfs and massive extrasolar planets are high enough to make them observable. PMID- 19129843 TI - Measured long-range repulsive Casimir-Lifshitz forces. AB - Quantum fluctuations create intermolecular forces that pervade macroscopic bodies. At molecular separations of a few nanometres or less, these interactions are the familiar van der Waals forces. However, as recognized in the theories of Casimir, Polder and Lifshitz, at larger distances and between macroscopic condensed media they reveal retardation effects associated with the finite speed of light. Although these long-range forces exist within all matter, only attractive interactions have so far been measured between material bodies. Here we show experimentally that, in accord with theoretical prediction, the sign of the force can be changed from attractive to repulsive by suitable choice of interacting materials immersed in a fluid. The measured repulsive interaction is found to be weaker than the attractive. However, in both cases the magnitude of the force increases with decreasing surface separation. Repulsive Casimir Lifshitz forces could allow quantum levitation of objects in a fluid and lead to a new class of switchable nanoscale devices with ultra-low static friction. PMID- 19129844 TI - Electrically pumped photonic-crystal terahertz lasers controlled by boundary conditions. AB - Semiconductor lasers based on two-dimensional photonic crystals generally rely on an optically pumped central area, surrounded by un-pumped, and therefore absorbing, regions. This ideal configuration is lost when photonic-crystal lasers are electrically pumped, which is practically more attractive as an external laser source is not required. In this case, in order to avoid lateral spreading of the electrical current, the device active area must be physically defined by appropriate semiconductor processing. This creates an abrupt change in the complex dielectric constant at the device boundaries, especially in the case of lasers operating in the far-infrared, where the large emission wavelengths impose device thicknesses of several micrometres. Here we show that such abrupt boundary conditions can dramatically influence the operation of electrically pumped photonic-crystal lasers. By demonstrating a general technique to implement reflecting or absorbing boundaries, we produce evidence that whispering-gallery like modes or true photonic-crystal states can be alternatively excited. We illustrate the power of this technique by fabricating photonic-crystal terahertz (THz) semiconductor lasers, where the photonic crystal is implemented via the sole patterning of the device top metallization. Single-mode laser action is obtained in the 2.55-2.88 THz range, and the emission far field exhibits a small angular divergence, thus providing a solution for the quasi-total lack of directionality typical of THz semiconductor lasers based on metal-metal waveguides. PMID- 19129845 TI - Early formation of evolved asteroidal crust. AB - Mechanisms for the formation of crust on planetary bodies remain poorly understood. It is generally accepted that Earth's andesitic continental crust is the product of plate tectonics, whereas the Moon acquired its feldspar-rich crust by way of plagioclase flotation in a magma ocean. Basaltic meteorites provide evidence that, like the terrestrial planets, some asteroids generated crust and underwent large-scale differentiation processes. Until now, however, no evolved felsic asteroidal crust has been sampled or observed. Here we report age and compositional data for the newly discovered, paired and differentiated meteorites Graves Nunatak (GRA) 06128 and GRA 06129. These meteorites are feldspar-rich, with andesite bulk compositions. Their age of 4.52 +/- 0.06 Gyr demonstrates formation early in Solar System history. The isotopic and elemental compositions, degree of metamorphic re-equilibration and sulphide-rich nature of the meteorites are most consistent with an origin as partial melts from a volatile-rich, oxidized asteroid. GRA 06128 and 06129 are the result of a newly recognized style of evolved crust formation, bearing witness to incomplete differentiation of their parent asteroid and to previously unrecognized diversity of early-formed materials in the Solar System. PMID- 19129846 TI - Diversity dynamics of marine planktonic diatoms across the Cenozoic. AB - Diatoms are the dominant group of phytoplankton in the modern ocean. They account for approximately 40% of oceanic primary productivity and over 50% of organic carbon burial in marine sediments. Owing to their role as a biological carbon pump and effects on atmospheric CO(2) levels, there is great interest in elucidating factors that influenced the rapid rise in diatom diversity during the past 40 million years. Two biotic controls on diversification have been proposed to explain this diversity increase: (1) geochemical coupling between terrestrial grasslands and marine ecosystems through the global silicon cycle; and (2) competitive displacement of other phytoplankton lineages. However, these hypotheses have not been tested using sampling-standardized fossil data. Here we show that reconstructions of species diversity in marine phytoplankton reject these proposed controls and suggest a new pattern for oceanic diatom diversity across the Cenozoic. Peak species diversity in marine planktonic diatoms occurred at the Eocene-Oligocene boundary and was followed by a pronounced decline, from which diversity has not recovered. Although the roles of abiotic and biotic drivers of diversification remain unclear, major features of oceanic diatom evolution are decoupled from both grassland expansion and competition among phytoplankton groups. PMID- 19129847 TI - Structural basis for androgen specificity and oestrogen synthesis in human aromatase. AB - Aromatase cytochrome P450 is the only enzyme in vertebrates known to catalyse the biosynthesis of all oestrogens from androgens. Aromatase inhibitors therefore constitute a frontline therapy for oestrogen-dependent breast cancer. In a three step process, each step requiring 1 mol of O(2), 1 mol of NADPH, and coupling with its redox partner cytochrome P450 reductase, aromatase converts androstenedione, testosterone and 16alpha-hydroxytestosterone to oestrone, 17beta oestradiol and 17beta,16alpha-oestriol, respectively. The first two steps are C19 methyl hydroxylation steps, and the third involves the aromatization of the steroid A-ring, unique to aromatase. Whereas most P450s are not highly substrate selective, it is the hallmark androgenic specificity that sets aromatase apart. The structure of this enzyme of the endoplasmic reticulum membrane has remained unknown for decades, hindering elucidation of the biochemical mechanism. Here we present the crystal structure of human placental aromatase, the only natural mammalian, full-length P450 and P450 in hormone biosynthetic pathways to be crystallized so far. Unlike the active sites of many microsomal P450s that metabolize drugs and xenobiotics, aromatase has an androgen-specific cleft that binds the androstenedione molecule snugly. Hydrophobic and polar residues exquisitely complement the steroid backbone. The locations of catalytically important residues shed light on the reaction mechanism. The relative juxtaposition of the hydrophobic amino-terminal region and the opening to the catalytic cleft shows why membrane anchoring is necessary for the lipophilic substrates to gain access to the active site. The molecular basis for the enzyme's androgenic specificity and unique catalytic mechanism can be used for developing next-generation aromatase inhibitors. PMID- 19129849 TI - Differential expression of SMAD3 transcripts is not regulated by cis-acting genetic elements but has a gender specificity. AB - As a key component of the transforming growth factor-beta (TGF-beta) pathway, SMAD3 plays an essential role in development and maintenance of self-tolerance. Furthermore, a recent study based on gene-expression profiling in donors of allogeneic hematopoietic cell grafts revealed that the level of expression of several components of the TGF-beta pathway can predict the occurrence of graft versus-host disease (GVHD) in recipients. The gene with the best GVHD predictive accuracy was SMAD3: no recipients suffered from GVHD when their donor cells expressed high levels of SMAD3 transcripts. The present study had two specific aims: to validate differential expression of SMAD3 transcripts in an independent and larger cohort of subjects and to determine whether interindividual differences were dictated by cis-acting genetic elements. In a cohort of 397 subjects, we found that SMAD3 transcript levels varied over a sixfold range. Analyses of SMAD3 single nucleotide polymorphisms and of SMAD3 promoter methylation patterns provide compelling evidence that interindividual differences in SMAD3 transcript levels do not result from in-cis genetic variations. Of note, part of the variance in SMAD3 expression was gender related as women expressed lower levels of SMAD3 transcripts than men. PMID- 19129850 TI - Molecular subtypes of systemic sclerosis in association with anti-centromere antibodies and digital ulcers. AB - The objective of this study was to identify molecular profiles that may distinguish clinical subtypes in systemic sclerosis (SSc). Large-scale gene expression profiling was performed on peripheral blood (PB) from 12 SSc patients and 6 healthy individuals. Significance analysis of microarrays, two-way hierarchical cluster analysis and PANTHER (Protein ANalysis THrough Evolutionary Relationships) ontology classification were used to analyze the data. Quantitative PCR was applied for validation in a cohort of 43 SSc patients. The results show that the expression of genes involved in immune defense, cell cycle and signal transduction was significantly elevated in PB of SSc patients (n=12) compared with healthy individuals (n=6). SSc patients could be stratified into subgroups based on differential expression of genes induced by type I interferon (IFN) and genes involved in antimicrobial (AM) activity. Differential expression of type I IFN or AM signature genes was validated and extended in an independent cohort of 31 patients by quantitative PCR. Low expression of IFN response genes was associated with the presence of anti-centromere antibodies, whereas increased expression was associated with the appearance of digital ulcers. In conclusion, patients with SSc can be classified on the basis of differential expression of immune defense genes. Differences in the activity of the type I IFN response program stratify patients into two clinically relevant subgroups. PMID- 19129851 TI - The impact of cell re-entry into the primary lymphoid organs on lymphocyte repertoire and functionality. PMID- 19129852 TI - Peripheral T cells in the thymus: have they just lost their way or do they do something? AB - In young adult mice, the thymus produces about a million newly formed T cells every day that colonize peripheral lymphoid tissues. Mostly regarded as a primary lymphoid organ only, the relationship between the thymus and peripheral lymphoid organs is considered unidirectional. However, this perception has been challenged by reports showing that peripheral lymphocytes, mostly T cells, can migrate back into the thymus. The presence of recirculating T cells in the thymus is rather incongruous and raises the question: is the presence of 'peripheral' T cells in the thymus superfluous or do these cells fulfill some relevant physiologic functions? There is now evidence that cells of the hematopoietic lineage, including T cells, can play an active role during thymocyte selection, a role generally considered the exclusive property of thymic epithelial cells and dendritic cells. Although, on a per cell basis, peripheral T cells in the thymus may be less efficient than thymus epithelial cells or dendritic cells at thymocyte positive and negative selection, they may nevertheless contribute to selection by influencing the selectable TCR repertoire and post-selection T cell functionality. Here, peripheral lymphocytes re-entering the thymus may be envisioned as Trojan horses as these cells may introduce antigens necessary for both positive and negative selection of T cells. PMID- 19129854 TI - Hypertension and obstructive sleep apnoea syndrome: current perspectives. AB - Obstructive sleep apnoea syndrome (OSAS), due to the collapse of the upper airways, is a common but still underestimated condition. The 'dose-response' type relationship between OSAS and hypertension (HT) has now been clearly proven. There are multiple mechanisms explaining this relationship, the main one being an increase in sympathetic activity during the apnoeas. HT associated with OSAS has several characteristics: high prevalence, diastolic and nocturnal predominance, and frequent non-dipper status. Furthermore, as OSAS is found in the majority of subjects with refractory HT, it should be systematically investigated in this situation. HT associated with OSAS should be tested for by means of a clinical blood pressure (BP) measurement, to which 24-h ambulatory BP monitoring (ABPM) is often added due to the fact that BP anomalies are frequently present at night. HT during OSAS is frequently associated with metabolic anomalies (for example, obesity, dyslipidaemia and insulin resistance), therefore explaining the high prevalence of metabolic syndrome in this population. The reference treatment for OSAS-nasal continuous positive airway pressure (nCPAP)-seems to be able to lower the BP of hypertensive patients, especially if the HT is severe, untreated or refractory. Moreover, the BP response to nCPAP depends on the severity of the OSAS, in particular the scale of the nocturnal desaturations, and on patient tolerance of the treatment. Optimal treatment for HT associated with OSAS has not been evidenced. Antihypertensive drugs do not change the respiratory parameters during OSAS. PMID- 19129855 TI - Home and office blood pressure in children and adolescents: the role of obesity. The Arsakeion School Study. AB - Obesity is related to office blood pressure (OBP). Important discrepancies exist between OBP and home blood pressure (HBP), providing complementary information for the management of hypertension. The association between obesity and HBP has not been investigated in children. The evidence on the role of obesity in the predominance of systolic blood pressure (SBP) over diastolic (DBP) in paediatric hypertension is limited. A total of 778 healthy subjects aged 6-18 years were recruited in this study. OBP and HBP were measured using electronic devices validated in children. Anthropometric measurements were measured and expressed as z-scores for height or age. Among all indices of obesity (z-scores), body mass index (BMI) showed the best association with BP. The effect of obesity (BMI) was more pronounced on: (i) SBP than DBP and (ii) H-SBP than O-SBP (O-SBP: r2=0.09, O DBP: r2=0.05, H-SBP: r2=0.12, H-DBP: r2=0.06). The prevalence of systolic hypertension was higher than that of diastolic hypertension. This difference was significant only in office readings and independent from obesity (normal weight: 6.3% systolic hypertension versus 1.2% diastolic; obese: 37.9% versus 6.9%, P<0.05 for both). These data imply that in children and adolescents the z-score of BMI is the most appropriate index of the association between BP and obesity. It also suggests that obesity is probably more closely associated with home than office BP. Finally, although obesity appears to affect SBP more than DBP, these results suggest that the predominance of systolic hypertension in children and adolescents might not be only related to obesity but also to the measurement setting (office). PMID- 19129856 TI - Frequency of renal artery stenosis and variants of renal vascularization in hypertensive patients: analysis of 1550 angiographies in one centre. AB - Renal artery stenosis (RAS) is an important cause of arterial hypertension and chronic kidney disease. The aims of our study were to assess the prevalence of RAS and to examine the frequency of variants of renal vasculature, that is, multiple and/or accessory renal arteries in hypertensive patients referred to renal angiography. We evaluated retrospectively 1554 arteriographies of hypertensive patients. Angiograms were evaluated to find RAS, significant RAS (>60% stenosis of the lumen), radiological signs of atherosclerosis, aneurysms of the renal arteries or aorta and variants of kidney vascularization. The frequency of RAS including occlusions was 15.1% (21.3% of them were significant and suitable for revascularization). Variants of renal arterial vascularization were found in 26.5% of patients (multiple renal arteries-11.2% and accessory renal arteries-15.3%). Significant RAS was found more frequently in patients older than 60 years-OR 4.76 (2.08-10.86). Coronary artery disease, history of myocardial infarction or stroke significantly increased the chance of RAS detection. The frequency of renal accessory arteries was lower in patients older than 60 years and in patients with the radiological signs of atherosclerosis. Results of this study indicate that haemodynamically important RAS is found more frequently in hypertensive patients older than 60 years. Symptomatic atherosclerotic disease found in the peripheral and/or coronary arteries and diabetes mellitus increases the chance of RAS detection. Decreased occurrence of renal accessory arteries was found in hypertensive patients with radiological signs of atherosclerosis. PMID- 19129857 TI - Clinical usefulness of the second peak of radial systolic blood pressure for estimation of aortic systolic blood pressure. AB - Central aortic blood pressure (BP), obtained from radial arterial waveform using the transfer function method (TFM), has been shown to have prognostic value independently of brachial BP. In this study, the relationship between peripheral systolic BP (SBP) and aortic SBP was evaluated. We further investigated whether TFM-derived aortic SBP can be estimated by information obtained from the radial waveform. The radial waveform was analysed to obtain the first peak of radial SBP (SBP1), second peak of radial SBP (SBP2), radial augmentation index (AI) (radial (SBP2-DBP)/(SBP1-DBP) x 100 and aortic SBP and AI using TFM in 233 subjects in the supine position. Measurements were repeated after changing position to the prone position. The constructed equation was validated in 149 community residents with different backgrounds. Radial SBP2 was closer to TFM-derived aortic SBP compared with brachial SBP. TFM-derived aortic SBP was approximated by the equation: aortic SBP=18.9-radial SBP2-0.03 x HR-0.214 x radial AI (r2=0.992). The equation was also applicable to predicting aortic SBP in the prone position as well as in different populations (mean difference between predicted aortic SBP and TFM-derived aortic SBP: -0.01+/-1.34 and 1.05+/-1.47 mm Hg, respectively). Radial arterial waveform analysis can be used for estimation of TFM-derived aortic SBP. PMID- 19129858 TI - Progress and prospects: the design and production of plasmid vectors. AB - Plasmid DNA (pDNA) expression vectors are fundamental to all forms of non-viral gene transfer. In this review, we discuss principles of pDNA design and production including the impact of bacterially derived sequences on transgene expression and minicircle approaches to minimize their effects. The impact of inclusion of DNA elements such as scaffold matrix attachment regions (S/MARs), transcription factor (TF)-binding sites and tissue-specific promoters are described. The benefits of eliminating CG dinucleotides (CpGs) from the pDNA are also considered. PMID- 19129859 TI - Identification of the murine firefly luciferase-specific CD8 T-cell epitopes. AB - In vivo bioluminescence imaging of reporter enzymes has proven to be a uniquely powerful tool that allows the study of the biology of viral and nonviral gene transfer agents. Cost-effective, noninvasive, longitudinal gene transfer studies in individual animals yield important information, which can influence the design of subsequent preclinical studies. The broad and expanding use of luciferase transgenes, specifically firefly luciferase, has prompted the study of luciferase specific T-cell activation following in vivo gene transfer. Herein, we report the mapping of the dominant T cell epitope in C57BL/6 mice (LMYRFEEEL) and the mapping of the dominant and minor T-cell epitopes in BALB/c mice (GFQSMYTFV and VPFHHGFGM, VALPHRTAC, respectively). These CD8 T-cell epitopes can be used to monitor cellular responses in vivo as well as be important tools in studies designed to suppress transgene-specific T cells. PMID- 19129860 TI - An adenoviral vector-based mucosal vaccine is effective in protection against botulism. AB - A replication-incompetent adenoviral vector encoding the heavy chain C-fragment (H(C)50) of botulinum neurotoxin type C (BoNT/C) was evaluated as a mucosal vaccine against botulism in a mouse model. Single intranasal inoculation of the adenoviral vector elicited a high level of H(C)50-specific IgG, IgG1 and IgG2a in sera and IgA in mucosal secretions as early as 2 weeks after vaccination. The antigen-specific serum antibodies were maintained at a high level at least until the 27th week. Immune sera showed high potency in neutralizing BoNT/C as indicated by in vitro toxin neutralization assay. The mice receiving single dose of 2 x 10(7) p.f.u. (plaque-forming unit) of adenoviral vector were completely protected against challenge with up to 10(4) x MLD(50) of BoNT/C. The protective immunity showed vaccine dose dependence from 10(5) to 2 x 10(7) p.f.u. of adenoviral vector. In addition, animals receiving single intranasal dose of 2 x 10(7) p.f.u. adenoviral vector could be protected against 100 x MLD(50) 27 weeks after vaccination. Animals with preexisting immunity to adenovirus could also be vaccinated intranasally and protected against lethal challenge with BoNT/C. These results suggest that the adenoviral vector is a highly effective gene-based mucosal vaccine against botulism. PMID- 19129861 TI - Gene delivery to human adult and embryonic cell-derived stem cells using biodegradable nanoparticulate polymeric vectors. AB - Gene delivery to stem cells holds great potential for tissue regeneration and delivery of therapeutic proteins. The major barrier is the lack of safe and efficient delivery methods. Here, we report enhanced gene delivery systems for human stem cells using biodegradable polymeric vectors. A library of poly (beta amino esters) end-modified derivatives was developed and optimized for high transfection efficiency and low cytotoxicity for three human stem cell lines including human mesenchymal stem cells (hMSCs), human adipose-derived stem cells (hADSCs) and human embryonic stem cell-derived cells (hESCds). In the presence of 10% serum, leading end-modified C32 polymeric vectors exhibited significantly high transfection efficiency in hMSCs (27+/-2%), hADSCs (24+/-3%) and hESCds (56+/-11%), with high cell viability (87-97%) achieved in all cell types. Our results show that poly(beta-amino esters) as a class, and end-modified versions of C32 in particular, are efficient polymeric vectors for gene delivery to both adult and embryonic-derived stem cells. PMID- 19129862 TI - Grazing rates and functional diversity of uncultured heterotrophic flagellates. AB - Aquatic assemblages of heterotrophic protists are very diverse and formed primarily by organisms that remain uncultured. Thus, a critical issue is assigning a functional role to this unknown biota. Here we measured grazing rates of uncultured protists in natural assemblages (detected by fluorescent in situ hybridization (FISH)), and investigated their prey preference over several bacterial tracers in short-term ingestion experiments. These included fluorescently labeled bacteria (FLB) and two strains of the Roseobacter lineage and the family Flavobacteriaceae, of various cell sizes, which were offered alive and detected by catalyzed reporter deposition-FISH after the ingestion. We obtained grazing rates of the globally distributed and uncultured marine stramenopiles groups 4 and 1 (MAST-4 and MAST-1C) flagellates. Using FLB, the grazing rate of MAST-4 was somewhat lower than whole community rates, consistent with its small size. MAST-4 preferred live bacteria, and clearance rates with these tracers were up to 2 nl per predator per h. On the other hand, grazing rates of MAST-1C differed strongly depending on the tracer prey used, and these differences could not be explained by cell viability. Highest rates were obtained using FLB whereas the flavobacteria strain was hardly ingested. Possible explanations would be that the small flavobacteria cells were outside the effective size range of edible prey, or that MAST-1C selects against this particular strain. Our original dual FISH protocol applied to grazing experiments reveals important functional differences between distinct uncultured protists and offers the possibility to disentangle the complexity of microbial food webs. PMID- 19129863 TI - Viral lysis of Phaeocystis pouchetii: implications for algal population dynamics and heterotrophic C, N and P cycling. AB - A model ecosystem with two autotrophic flagellates, Phaeocystis pouchetii and Rhodomonas salina, a virus specific to P. pouchetii (PpV) and bacteria and heterotrophic nanoflagellates was used to investigate effects of viral lysis on algal population dynamics and heterotrophic nitrogen and phosphorus mineralization. Lysis of P. pouchetii by PpV had strong positive effects on bacterial and HNF abundance, and the mass balance of C, N and P suggested an efficient transfer of organic material from P. pouchetii to bacterial and HNF biomass through viral lysis. At the same time, the degradation of P. pouchetii lysates was associated with significant regeneration of inorganic N and P resulting in 148 microg N l(-1) and 7 microg P l(-1), corresponding to 78% and 26% of lysate N and P being mineralized to NH(4)(+) and PO(4)(3-), respectively. These results showed that the turnover of viral lysates in the microbial food web was associated with significant N and P mineralization, supporting the current view that viral lysates can be an important source of inorganic nutrients in marine systems. In the presence of R. salina, the generated NH(4)(+) supported 11% of the observed R. salina growth. Regrowth of virus-resistant P. pouchetii following cell lysis was observed in long-term incubations (150 days), and possibly influenced by nutrient availability and competition from R. salina. The observed impact of viral activity on autotrophic and heterotrophic processes provides direct experimental evidence for virus-driven nutrient generation and emphasizes the potential importance of the viral activity in supporting marine primary production. PMID- 19129865 TI - Quantifying expression of Geobacter spp. oxidative stress genes in pure culture and during in situ uranium bioremediation. AB - As part of an effort to diagnose the physiological status of Geobacter species during in situ bioremediation of uranium-contaminated groundwater, transcript levels for two genes potentially associated with oxidative stress, cydA and sodA, were quantified throughout a bioremediation field study in Rifle, CO, USA. Despite the accumulation of Fe(II) in the groundwater, which is inconsistent with the presence of dissolved oxygen, both genes were highly expressed during the bioremediation process. Therefore, the response to oxidative stress was further evaluated with Geobacter uraniireducens, an isolate from the Rifle site. When G. uraniireducens cultured with fumarate as the electron acceptor was exposed to 5% oxygen for 8 h, there was a significant increase in cydA and sodA transcripts as well as other genes associated with oxygen respiration or oxidative stress. Oxygen-exposed cells had lower transcript abundance for genes associated with anaerobic respiration, metabolism and motility. Short-term oxygen exposure had little impact on cydA transcript levels, as more than 1 h was required for increases to levels comparable to the subsurface. Abundance of cydA and sodA transcripts for the isolate G. sulfurreducens were always higher in cells cultured with Fe(III) compared with fumarate as an electron acceptor, even when fumarate-grown cells were exposed to oxygen, and Fe(III)-grown cells were grown anaerobically. These results suggest that the apparently high Geobacter cydA and sodA expression during bioremediation cannot necessarily be attributed to oxidative stress and demonstrate that diagnosis of the metabolic status of subsurface microorganisms through transcript analysis should be coupled with appropriate geochemical analyses. PMID- 19129867 TI - Surface-emitting circular DFB, disk-, and ring-Bragg resonator lasers with chirped gratings. II: nonuniform pumping and far-field patterns. AB - This is a continuation of our previous work [Opt. Express 16, 9155 (2008)]. In this paper we investigate the effect of nonuniform pumping on the modal properties of surface-emitting chirped circular grating lasers. By numerically solving the coupled-mode equations and matching the boundaries we compare and discuss the threshold pump levels and frequency detuning factors for three pumping profiles: uniform, Gaussian, and annular. Depending on the overlap of the pumping and modal profiles, Gaussian pumping results in the lowest threshold pump levels except for the fundamental mode of ring Bragg resonator laser, and annular pumping provides larger threshold discrimination between the fundamental and first-order modes of circular DFB and ring Bragg resonator lasers, which is favorable for single-mode operation in these lasers. We also study the far-field patterns of the fundamental modes of circular DFB, disk-, and ring- Bragg resonator lasers. Circular DFB and ring Bragg resonator lasers have the first order dominating peak, while disk Bragg resonator laser exhibits the zeroth-order dominating peak. PMID- 19129868 TI - Dispersion encoded full range frequency domain optical coherence tomography. AB - We propose an iterative algorithm that exploits the dispersion mismatch between reference and sample arm in frequency-domain optical coherence tomography (FD OCT) to effectively cancel complex conjugate mirror terms in individual A-scans and thereby generate full range tomograms. The resulting scheme, termed dispersion encoded full range (DEFR) OCT, allows distinguishing real structures from complex conjugate mirror artifacts. Even though DEFR-OCT has higher post processing complexity than conventional FD-OCT, acquisition speed is not compromised since no additional A-scans need to be measured, thereby rendering this technique robust against phase fluctuations. The algorithm uses numerical dispersion compensation and exhibits similar resolution as standard processing. The residual leakage of mirror terms is further reduced by incorporating additional knowledge such as the power spectrum of the light source. The suppression ratio of mirror signals is more than 50 dB and thus comparable to complex FD-OCT techniques which use multiple A-scans. PMID- 19129869 TI - Optical arbitrary waveform characterization via dual-quadrature spectral interferometry. AB - We introduce the use of dual-quadrature spectral interferometry for amplitude and phase characterization of 100% duty factor optical arbitrary waveforms generated via spectral line-by-line pulse shaping. We demonstrate this technique for measurement of optical arbitrary waveforms composed of approximately 30 spectral lines from a 10 GHz frequency comb with 1.4 micros data acquisition time at an average power level of 10 microwatts. We then demonstrate coherent spectral phase measurements of pulses strongly dispersed by propagation over 50 km of optical fiber. PMID- 19129870 TI - Electromagnetic fields and transmission properties in tapered hollow metallic waveguides. AB - We analyze the electromagnetic spatital distributions and address an important issue of the transmission properties of spherical transverse-electric (TE) and transverse-magnetic (TM) eigenmodes within a tapered hollow metallic waveguide in detail. Explicit analytical expressions for the spatital distributions of electromagnetic field components, attenuation constant, phase constant and wave impedance are derived. Accurate eigenvalues obtained numerically are used to study the dependences of the transmission properties on the taper angle, the mode as well as the length of the waveguide. It is shown that all modes run continuously from a propagating through a transition to an evanescent region and the value of the attenuation increases as the distance from the cone vertex and the cone angle decrease. A strict distinction between pure propagating and pure evanescent modes cannot be achieved. One mode after the other reaches cutoff in the tapered hollow metallic waveguide as the distance from the cone vertex desreases. PMID- 19129871 TI - Periodic metallo-dielectric structure in diamond. AB - Intense ultrashort light pulses induce three dimensional localized phase transformation of diamond. Photoinduced amorphous structures have electrical conducting properties of a maximum of 64 S/m based on a localized transition from sp(3) to sp(2) in diamond. The laser parameters of fluence and scanning speed affect the resultant electrical conductivities due to recrystallization and multi filamentation phenomena. We demonstrate that the laser-processed diamond with the periodic cylinder arrays have the characteristic transmission properties in terahertz region, which are good agreement with theoretical calculations. The fabricated periodic structures act as metallo-dielectric photonic crystal. PMID- 19129872 TI - Measuring temporal speckle correlations at ultrafast x-ray sources. AB - We present a new method to extract the intermediate scattering function from series of coherent diffraction patterns taken with 2D detectors. Our approach is based on analyzing speckle patterns in terms of photon statistics. We show that the information obtained is equivalent to the conventional technique of calculating the intensity autocorrelation function. Our approach represents a route for correlation spectroscopy on ultrafast timescales at X-ray free-electron laser sources. PMID- 19129873 TI - Generation of carrier-envelope-phase-stable 2-cycle 740-microJ pulses at 2.1 microm carrier wavelength. AB - We produce carrier-envelope-phase-stable 15.7-fs (2-cycle) 740-microJ pulses at the 2.1-microm carrier wavelength, from a three-stage optical parametric chirped pulse amplifier system, pumped by an optically synchronized 49-ps 11-mJ Nd:YLF laser. A novel seed pulse spectral shaping method is used to ascertain the true amplified seed energy and the parametric superfluorescence levels. PMID- 19129874 TI - A simple perturbative analysis for fast design of an electrically pumped micro disk laser. AB - A perturbative analysis is proposed to estimate optical losses for electrically pumped micro-disk lasers. The optical field interaction with the electrical contacts and the optimization of their implementation is investigated. Our model shows a good agreement with 3D Finite Difference Time Domain (FDTD) computation and can be used for designing contacts for thin micro-disks, with a considerably reduced calculation time. We also demonstrate that losses induced by the contacts can be exploited to select the optical mode of a micro-laser. PMID- 19129864 TI - A comprehensive survey of soil acidobacterial diversity using pyrosequencing and clone library analyses. AB - Acidobacteria are ubiquitous and abundant members of soil bacterial communities. However, an ecological understanding of this important phylum has remained elusive because its members have been difficult to culture and few molecular investigations have focused exclusively on this group. We generated an unprecedented number of acidobacterial DNA sequence data using pyrosequencing and clone libraries (39,707 and 1787 sequences, respectively) to characterize the relative abundance, diversity and composition of acidobacterial communities across a range of soil types. To gain insight into the ecological characteristics of acidobacterial taxa, we investigated the large-scale biogeographic patterns exhibited by acidobacterial communities, and related soil and site characteristics to acidobacterial community assemblage patterns. The 87 soils analyzed by pyrosequencing contained more than 8600 unique acidobacterial phylotypes (at the 97% sequence similarity level). One phylotype belonging to Acidobacteria subgroup 1, but not closely related to any cultured representatives, was particularly abundant, accounting for 7.4% of bacterial sequences and 17.6% of acidobacterial sequences, on average, across the soils. The abundance of Acidobacteria relative to other bacterial taxa was highly variable across the soils examined, but correlated strongly with soil pH (R= 0.80, P<0.001). Soil pH was also the best predictor of acidobacterial community composition, regardless of how the communities were characterized, and the relative abundances of the dominant Acidobacteria subgroups were readily predictable. Acidobacterial communities were more phylogenetically clustered as soil pH departed from neutrality, suggesting that pH is an effective habitat filter, restricting community membership to progressively more narrowly defined lineages as pH deviates from neutrality. PMID- 19129866 TI - Bacterial diversity and White Plague Disease-associated community changes in the Caribbean coral Montastraea faveolata. AB - Increasing evidence confirms the crucial role bacteria and archaea play within the coral holobiont, that is, the coral host and its associated microbial community. The bacterial component constitutes a community of high diversity, which appears to change in structure in response to disease events. In this study, we highlight the limitation of 16S rRNA gene (16S rDNA) clone library sequencing as the sole method to comprehensively describe coral-associated communities. This limitation was addressed by combining a high-density 16S rRNA gene microarray with, clone library sequencing as a novel approach to study bacterial communities in healthy versus diseased corals. We determined an increase in diversity as well as a significant shift in community structure in Montastraea faveolata colonies displaying phenotypic signs of White Plague Disease type II (WPD-II). An accumulation of species that belong to families that include known coral pathogens (Alteromonadaceae, Vibrionaceae), bacteria previously isolated from diseased, stressed or injured marine invertebrates (for example, Rhodobacteraceae), and other species (for example, Campylobacteraceae) was observed. Some of these species were also present in healthy tissue samples, but the putative primary pathogen, Aurantimonas corallicida, was not detected in any sample by either method. Although an ecological succession of bacteria during disease progression after causation by a primary agent represents a possible explanation for our observations, we also discuss the possibility that a disease of yet to be determined etiology may have affected M. faveolata colonies and resulted in (or be a result of) an increase in opportunistic pathogens. PMID- 19129876 TI - Cascaded optical parametric oscillations generating tunable terahertz waves in periodically poled lithium niobate crystals. AB - We present a continuous-wave (cw) singly-resonant optical parametric oscillator (SROPO) based on MgO-doped periodically poled lithium niobate (PPLN) delivering single-frequency idler output from 2.33 to 5.32 microm. In this system, we observe additional spectral components that have been attributed to stimulated Raman lines in other studies. However, we are able to assign them unambiguously to cascaded optical parametric processes. The tunable forward and backward idler waves generated by these additional phase-matched oscillations have frequencies that are tunable around 3.5 and 1.5 THz, respectively. PMID- 19129875 TI - Nanofabricated upconversion nanoparticles for photodynamic therapy. AB - We present a novel process for the production of three-layer Composite Nanoparticles (CNPs) in the size range 100-300 nm with an up-converting phosphor interior, a coating of porphyrin photosensitizer, and a biocompatible PEG outer layer to prevent clearance by the reticuloendothelial system. We show that these CNPs produce millimolar amounts of singlet oxygen at NIR intensities far less than other two-photon techniques. PMID- 19129877 TI - Speckle suppression in laser display using several partially coherent beams. AB - An optical scheme for speckle suppression using two or three partially coherent beams in a projection system is proposed. Diffractive optical elements (DOE) placed in the intermediate image plane create several beams carrying the image to a screen. Transparent plates of different thicknesses are placed in the Fourier plane of the projective lens and used for beam decorrelation. The coherence matrix algorithm for speckle suppression is used to calculate the speckle contrast ratio. It is shown that for a small decorrelation length and using the same maximum thickness of the transparent plates, two partially coherent beams would provide better suppression than three beams with different diffraction orders. However, for a large decorrelation length, the three beam setup provides better speckle suppression for all three colors examined with a suppression coefficient close to theoretical limits. Verification of speckle suppression using three-beam decorrelation is reported. PMID- 19129878 TI - Study on the decay mechanisms of surface plasmon coupling features with a light emitter through time-resolved simulations. AB - The transient behaviors of the dipole coupling with surface plasmon (SP) features in an Ag/dielectric-interface grating structure in order to understand the characteristics of those dipole-coupling features are demonstrated. In particular, the major decay mechanisms of those coupling features can be identified. For comparison, the time-resolved behaviors of the resonant surface plasmon polariton (SPP) coupling feature on a flat interface are also illustrated. Among the three major grating-induced SP-dipole coupling features, two of them are identified to be localized surface plasmons (LSPs). The third one is a grating-assisted SPP, which shows two decay components, corresponding to the first stage of SPP in-plane propagation and the second stage of coupling system decay. In all the dipole coupling features, metal dissipation can dominate the energy relaxation process, depending on the assumption of damping factor. All the dissipation rates are proportional to the assumed damping factor in the Drude model of the metal. The dissipation rates of the LSP and resonant SPP features are about the same as the damping rate, implying their local electron oscillation natures. The dissipation rate of the grating-assisted SSP feature is consistent with theoretical calculation. In the LSP features under study, dielectric-side emission is prominent. The coupled energy in the grating-assisted SPP feature can be efficiently stored in the coupling system due to its low emission efficiency and effective energy confinement through grating diffraction. PMID- 19129879 TI - Tailoring the excitation of localized surface plasmon-polariton resonances by focusing radially-polarized beams. AB - We study the interaction of focused radially-polarized light with metal nanospheres. By expanding the electromagnetic field in terms of multipoles, we gain insight on the excitation of localized surface plasmon-polariton resonances in the nanoparticle. We show that focused radially-polarized beams offer more opportunities than a focused plane wave or a Gaussian beam for tuning the near- and far-field system response. These results find applications in nano-optics, optical tweezers, and optical data storage. PMID- 19129880 TI - Single-shot interferometric approach to background free broadband coherent anti Stokes Raman scattering spectroscopy. AB - We introduce a single-shot interferometric approach to suppress the nonresonant background (NRB) contribution to a broadband coherent anti-Stokes Raman scattering (CARS) spectrum; this single-shot approach is conducive to rapid imaging. A pulse shaper prepares a narrowband pulse with two spectral components of differing phase. When the CARS fields generated by these two out-of-phase components are optically mixed, NRB signal is greatly reduced while a resonant CARS signal remains with minimal attenuation. We discuss and demonstrate two model schemes for the interfering pulse components: (1) two pulses with different bandwidths and the same center frequency (ps-fs scheme) and (2) two pulses with the same bandwidth and shifted center frequencies (ps-ps scheme). In both schemes, only the resonant signal from the "3-color" CARS mechanism survives. The resonant signal from "2-color" CARS mechanism vanishes along with the NRB. We discuss optimization conditions for signal intensity and shape of resonant CARS peaks. Experimental CARS spectra of c-hexane and benzonitrile demonstrate feasibility of these approaches. PMID- 19129882 TI - Chip-scale hybrid optical sensing systems using digital signal processing. AB - We propose a novel hybrid optical sensing system for standalone, chip-scale sensing applications. The hybrid optical sensing system detects any spectral shift of the microresonator sensor output by estimating the effective refractive index using maximum likelihood estimation. The performance evaluation of the proposed hybrid sensing system in the Gaussian-noise dominant environment shows excellent estimation accuracy. This innovative approach allows fully functional integrated hybrid sensing systems, offering great potential in various chip-scale sensing applications. PMID- 19129881 TI - Comparison of birefringent electric split-ring resonator and meanderline structures as quarter-wave plates at terahertz frequencies. AB - We have fabricated a quarter-wave plate from a single layer of birefringent electric split-ring resonators (ELC). For comparison, an appropriately scaled double layer meanderline structure was fabricated. At the design frequency of 639 GHz, the ELC structure achieves 99.9% circular polarization while the meanderline achieves 99.6%. The me-anderline displays a larger bandwidth of operation, attaining over 99% circular polarization from 615 - 743 GHz, while the ELC achieves 99% from 626 - 660 GHz. However, both are broad enough for use with CW sources making ELCs a more attractive choice due to the ease of fabrication. Both samples are free standing with a total thickness of 70 microm for the meanderline structure and a mere 20 microm for the ELC highlighting the large degree of birefringence exhibited with metamaterial structures. PMID- 19129883 TI - Electroluminescent devices based on amorphous SiN/Si quantum dots/amorphous SiN sandwiched structures. AB - A single layer of dense Si quantum dots with average size of 4 nm sandwiched in amorphous SiN layers was prepared by laser crystallization of ultrathin amorphous Si film followed by subsequently thermal annealing. The electroluminescent diodes were fabricated by evaporating Al electrodes on back sides of p-Si substrates and the top surface of samples. Room temperature electroluminescence can be detected with applying the negative voltage around 10V on the top gate electrode and the luminescent intensity is increased with increasing the applied voltage. It was found that the integrated luminescent intensity is linearly proportional to the injection current which suggested the intensity depends on the concentrations of injected carriers after Fowler-Nordheim tunneling through amorphous SiN barriers. The influence of the amorphous SiN with different band gap on the device performance was also discussed briefly. PMID- 19129884 TI - Demonstration of a cavity coupler based on a resonant waveguide grating. AB - Thermal noise in multilayer optical coatings may not only limit the sensitivity of future gravitational wave detectors in their most sensitive frequency band but is also a major impediment for experiments that aim to reach the standard quantum limit or to cool mechanical systems to their quantum ground state. Here, we present the experimental realization and characterization of a cavity coupler, which is based on a surface relief guided ode resonant grating. Since the required thickness of the dielectric coating is dramatically decreased compared to conventional mirrors, it is expected to provide low mechanical loss and, thus, low thermal noise. The cavity coupler was incorporated into a Fabry-Perot resonator together with a conventional high quality mirror. The finesse of this cavity was measured to be F = 657, which corresponds to a coupler reflectivity of R = 99.08 %. PMID- 19129885 TI - Stopping light by an air waveguide with anisotropic metamaterial cladding. AB - We present a detailed study of oscillating modes in a slab waveguide with air core and anisotropic metamaterial cladding. It is shown that, under specific dielectric configurations, slow and even stopped electromagnetic wave can be supported by such an air waveguide. We propose a linearly tapped waveguide structure that could lead the propagating light to a complete standstill. Both the theoretical analysis and the proposed waveguide have been validated by full wave simulation based on finite-difference time-domain method. PMID- 19129886 TI - Investigation of the thermally induced laser beam distortion associated with vacuum compressor gratings in high energy and high average power femtosecond laser systems. AB - We report successful compensation of the thermally induced laser beam distortion associated with high energy 110 mJ and high average power femtosecond laser system of 11 Watts operated with vacuum compressor gratings. To enhance laser based light source brightness requires development of laser systems with higher energy and higher average power. Managing the high thermal loading on vacuum optical components is a key issue in the implementation of this approach. To our knowledge this is the first time that such thermal induced distortions on the vacuum compressor gratings are characterized and compensated. PMID- 19129887 TI - Plasmon-enhanced emission from optically-doped MOS light sources. AB - We evaluate the spontaneous emission rate (Purcell) enhancement for optically doped metal-dielectric-semiconductor light-emitting structures by considering the behavior of a semiclassical oscillating point dipole placed within the dielectric layer. For a Ag-SiO(2)-Si structure containing emitters at the center of a 20-nm thick SiO(2) layer, spontaneous emission rate enhancements of 40 to 60 can be reached in the wavelength range of 600 to 1800 nm, far away from the surface plasmon resonance; similar enhancements are also possible if Al is used instead of Ag. For dipoles contained in the thin oxide layer of a Ag-SiO(2)-Si-SiO(2) structure, the emission exhibits strong preferential coupling to a single well defined Si waveguide mode. This work suggests a means of designing a new class of power-efficient, high-modulation-speed, CMOS-compatible optical sources that take full advantage of the excellent electrical properties and plasmon-enhanced op cal properties afforded by MOS devices. PMID- 19129888 TI - Imaging scatterometry of butterfly wing scales. AB - We describe an imaging scatterometer allowing hemispherical reflectance measurements as a function of the angle of incidence. The heart of the scatterometer is an ellipsoidal reflector, which compresses the hemispherical reflection into a cone-shaped beam that can be imaged by a normal optical system. The instrument's performance is illustrated by measurements of the scattering profiles of the blue-iridescent dorsal wing scales of the nymphalid Morpho aega and the matte-green ventral wing scales of the lycaenid Callophrys rubi. PMID- 19129889 TI - Multi-wavelength laser generation with Bismuthbased Erbium-doped fiber. AB - A multi-wavelength laser comb is demonstrated using a nonlinear effect in a backward pumped Bismuth-based Erbium-doped fiber (Bi-EDF) for the first time. It uses a ring cavity resonator scheme containing a 215 cm long highly nonlinear Bi EDF, optical isolators, polarisation controller and 10 dB output coupler. The laser generates more than 10 lines of optical comb with a line spacing of approximately 0.41 nm at 1615.5 nm region using 146 mW of 1480 nm pump power. PMID- 19129890 TI - Non-thermal desorption/ablation of molecular solids induced by ultra-short soft x ray pulses. AB - We report the first observation of single-shot soft x-ray laser induced desorption occurring below the ablation threshold in a thin layer of poly (methyl methacrylate)--PMMA. Irradiated by the focused beam from the Free-electron LASer in Hamburg (FLASH) at 21.7 nm, the samples have been investigated by atomic-force microscope (AFM) enabling the visualization of mild surface modifications caused by the desorption. A model describing non-thermal desorption and ablation has been developed and used to analyze single-shot imprints in PMMA. An intermediate regime of materials removal has been found, confirming model predictions. We also report below-threshold multiple-shot desorption of PMMA induced by high-order harmonics (HOH) at 32 nm. Short-time exposure imprints provide sufficient information about transverse beam profile in HOH's tight focus whereas long-time exposed PMMA exhibits radiation-initiated surface ardening making the beam profile measurement infeasible. PMID- 19129891 TI - Nanodroplet real-time PCR system with laser assisted heating. AB - We report the successful application of low-power (approximately 30 mW) laser radiation as an optical heating source for high-speed real-time polymerase chain reaction (PCR) amplification of DNA in nanoliter droplets dispersed in an oil phase. Light provides the heating, temperature measurement, and Taqman real-time readout in nanoliter droplets on a disposable plastic substrate. A selective heating scheme using an infrared laser appears ideal for driving PCR because it heats only the droplet, not the oil or plastic substrate, providing fast heating and completing the 40 cycles of PCR in 370 seconds. No microheaters or microfluidic circuitry were deposited on the substrate, and PCR was performed in one droplet without affecting neighboring droplets. The assay performance was quantitative and its amplification efficiency was comparable to that of a commercial instrument. PMID- 19129892 TI - Low bending loss metal waveguide embedded in a free-standing multilayered polymer film. AB - Very low vertical bending loss is demonstrated in a flexible metal waveguide. The waveguide consists of an 8 nm-thick and 68 mm-long Ag strip embedded in a free standing multilayered low-loss polymer film. The polymer film is composed of a 10 microm-thick inner cladding with a refractive index of 1.524, and a pair of 20 microm-thick outer claddings which both have a refractive index of 1.514, resulting in a total thickness of 50 microm. The measured vertical bending loss is lower than 0.3 dB/180 masculine at a wavelength of 1310 nm for the bending radii down to 2 mm. PMID- 19129893 TI - 3-Dimensional heat analysis in short-length Er3+/Yb3+ co-doped phosphate fiber laser with upconversion. AB - A 3-dimenstional (3D) heat flow model of laser diode (LD) pumped short-length Er(3+)/Yb(3+) heavily co-doped phosphate fiber laser that includes the energy transfer upconversion (ETU) effects has been developed. The fully 3D analytical solution with the consideration of longitudinal heat flow has been given to describe the temperature distribution in phosphate fiber. The calculated results show that both ETU processes and longitudinal heat conduction have a great influence on the fiber laser performance. Finally, we have validated the analytical expression by measuring the temperature distribution of an end-pumped short-length Er(3+)/Yb(3+) co-doped fiber laser, which was placed into the copper tube. PMID- 19129894 TI - High-resolution broadband (>100 cm-1) infrared heterodyne spectro-radiometry using an external cavity quantum cascade laser. AB - Broadband thermal infrared heterodyne spectro-radiometry using an external cavity quantum cascade laser as a tunable local oscillator has been performed over a frequency range of more than 100 cm(-1) at a central frequency of 1190 cm(-1). Heterodyne spectro-radiometry is demonstrated for two local oscillator tuning modes: broadband tuning for transmission and emission spectroscopy of broadband absorbers (Freon 12), and broadband frequency selection in combination with fine continuous frequency tuning for high-resolution (0.021 cm(-1)) transmission spectroscopy (N(2)O). In each case concentration retrievals are performed and analyzed. The spectroradiometer noise level is demonstrated to be twenty two and eight times the fundamental shot-noise limit in the two scanning modes respectively. PMID- 19129895 TI - Design and analysis of two-dimensional high-index-contrast grating surface emitting lasers. AB - The use of a two-dimensional (2D) high-index-contrast grating (HCG) with square periodic lattice is proposed to realize surface-emitting lasers. This is possible because the use of 2D HCG, in which multiple resonant leaky modes are excited by the 2 orthogonal directions of the grating, causes the high reflective zone to be split into two regions. Hence, a dip of the reflectivity is formed to support the excitation of a resonant cavity-mode inside the 2D HCG. With suitable design on the dimensions of the 2D HCGs, Q factor as high as 1032 can be achieved. PMID- 19129896 TI - Optical diffraction tomography for high resolution live cell imaging. AB - We report the experimental implementation of optical diffraction tomography for quantitative 3D mapping of refractive index in live biological cells. Using a heterodyne Mach-Zehnder interferometer, we record complex field images of light transmitted through a sample with varying directions of illumination. To quantitatively reconstruct the 3D map of complex refractive index in live cells, we apply optical diffraction tomography based on the Rytov approximation. In this way, the effect of diffraction is taken into account in the reconstruction process and diffraction-free high resolution 3D images are obtained throughout the entire sample volume. The quantitative refractive index map can potentially serve as an intrinsic assay to provide the molecular concentrations without the addition of exogenous agents and also to provide a method for studying the light scattering properties of single cells. PMID- 19129897 TI - Isolation and one-way effects in diffraction on dielectric gratings with plasmonic inserts. AB - Diffraction of plane waves on dielectric gratings with planar plasmonic inserts is studied with the emphasis put on the anomalous selectivity of diffraction orders. It is shown that some formally propagating orders can be suppressed within a wide frequency range. The effect of suppression is more general than the isolation effect observed earlier in zero-permittivity and (near-)zero-index slabs and sensitive to the frequency dependent peculiarities of the field distribution within the plasmonic layer. It is required that the real part of the permittivity of this layer is positive less than unity. The wideband features of the suppression effect, i.e., one-way transmission and diffraction-free reflection are demonstrated. Narrowband selectivity effects are also studied. The structures suggested can be used for extending the potential of technologies that are based on multibeam operation and field transformation. PMID- 19129898 TI - Surface Plasmon Resonance sensor showing enhanced sensitivity for CO2 detection in the mid-infrared range. AB - We present the first optical sensor based on Surface Plasmon Resonance (SPR) operating in the mid-infrared range. The experimental setup is based on a Kretschmann geometry with Ti/Au layers deposited on a CaF(2) prism where light excitation is provided by a Quantum Cascade Laser (QCL) source. Evidence of SPR is presented and the sensing capability of the system is demonstrated by using CO(2) and N(2) mixtures as test samples. Due to the absorption of CO(2) at this wavelength, it is shown that the sensitivity of this configuration is five times higher than a similar SPR sensor operating in the visible range of the spectrum. PMID- 19129899 TI - Electro-optical modulator in a polymerinfiltrated silicon slotted photonic crystal waveguide heterostructure resonator. AB - We present a novel concept of a compact, ultra fast electro-optic modulator, based on photonic crystal resonator structures that can be realized in two dimensional photonic crystal slabs of silicon as core material employing a nonlinear optical polymer as infiltration and cladding material. The novel concept is to combine a photonic crystal heterostructure cavity with a slotted defect waveguide. The photonic crystal lattice can be used as a distributed electrode for the application of a modulation signal. An electrical contact is hence provided while the optical wave is kept isolated from the lossy metal electrodes. Thereby, well known disadvantages of segmented electrode designs such as excessive scattering are avoided. The optical field enhancement in the slotted region increases the nonlinear interaction with an external electric field resulting in an envisaged switching voltage of approximately 1 V at modulation speeds up to 100 GHz. PMID- 19129900 TI - Instabilities and fractal basins of attraction in optical system optimization. AB - Many optical design programs use various forms of the damped least-squares method for local optimization. In this paper, we show that damped least-squares algorithms, with maximized computational speed, can create sensitivity with respect to changes in initial conditions. In such cases, starting points, which are very close to each other, lead to different local minima after optimization. Computations of the fractal capacity dimension show that sets of these starting points, which lead to the same minimum (the basins of attraction for that minimum), have a fractal structure. Introducing more damping makes the optimization process stable. PMID- 19129901 TI - Near field detector for integrated surface plasmon resonance biosensor applications. AB - Integrated surface plasmon resonance biosensors promise to enable compact and portable biosensing at high sensitivities. To replace the far field detector traditionally used to detect surface plasmons we integrate a near field detector below a functionalized gold film. The evanescent field of a surface plasmon at the aqueous-gold interface is converted into photocurrent by a thin film organic heterojunction diode. We demonstrate that use of the near field detector is equivalent to the traditional far field measurement of reflectivity. The sensor is stable and reversible in an aqueous environment for periods of 6 hrs. For specific binding of neutravidin, the detection limit is 4 microg/cm(2). The sensitivity can be improved by reducing surface roughness of the gold layers and optimization of the device design. From simulations, we predict a maximum sensitivity that is two times lower than a comparable conventional SPR biosensor. PMID- 19129902 TI - Electromagnetic wave propagation in a Ag nanoparticle-based plasmonic power divider. AB - In this paper a new silver (Ag) nanoparticle-based structure is presented which shows potential as a device for front end applications, in nano-interconnects or power dividers. A novel oxide bar ensures waveguiding and control of the signal strength with promising results. The structure is simulated by the two dimensional finite difference time domain (FDTD) method considering TM polarization and the Drude model. The effect of different wavelengths, material loss, gaps and particle sizes on the overall performance is discussed. It is found that the maximum signal strength remains along the Ag metallic nanoparticles and can be guided to targeted end points. PMID- 19129903 TI - Slow and fast light in optical fibers using acoustooptic coupling between two co propagating modes. AB - We demonstrate numerically that acoustooptic interaction between two co propagating modes in an optical fiber can be utilized to obtain optical delays. Both positive and negative delays of several pulse lengths can be obtained. Based on the simulations we consider relevant experimental parameters. PMID- 19129904 TI - Measured luminance and visual appearance of multi-color persistent phosphors. AB - In the past decade, several persistent phosphors were intensively studied, such as CaAl(2)O(4):Eu,Nd (blue) and SrAl(2)O(4):Eu,Dy (green). Yellow-to-red emitting persistent phosphors have also been identified. All these phosphors are normally evaluated by measuring their afterglow as a function of time in photometric units. However, the eye sensitivity curve shifts from photopic vision to scotopic vision upon decreasing light intensity, with a corresponding decrease in red sensitivity. Performance of red-emitting persistent phosphors thus cannot be described with usual photometry. An alternative description is presented, based on recent vision research in the intermediate (mesopic) region and using a unified system of photometry. It turns out that this effect seriously compromises the development of red-emitting persistent phosphors. This is illustrated on a number of commercially available persistent phosphors. PMID- 19129905 TI - Self-imaging in periodic dielectric waveguides. AB - Self-imaging phenomena in periodic dielectric waveguides has been predicted and investigated based on multimode interference effect by using the plane wave expansion method and the finite-difference timedomain method. Asymmetric and symmetric interferences were discussed and respective imaging positions were calculated. As examples of application, a demultiplexer and a filter with ultracompact and simple structures were designed and demonstrated theoretically for optical communication wavelengths. PMID- 19129906 TI - Marijuana Treatment Entries Did Not Decrease After Aggressive Arrest Policies Were Implemented in New York City. AB - In the mid-late 1990s Mayor Rudy Giuliani and Police Chief William Bratton focused on arresting and detaining people for crimes that contributed to a lower "quality-of-life" in New York City. This aggressive arrest policy (AAP) resulted in a record growth in marijuana arrests. In 1992, the number of marijuana arrests was around 5,000. By 2000, the arrest rate hit an all-time high of about 60,000 (the large majority of which were for misdemeanor arrests in both years). Through a triangulation of data sources, including the Uniform Crime Reports and the Treatment Episode Data Set from 1992 to 2003, and other published accounts, this paper shows that entries into treatment for marijuana dramatically increased in New York City at the same time as misdemeanor and felony arrests for marijuana also rose. While it is unclear if these arrests caused the treatment increase (vis-a-vis criminal justice referral programs), the presence of these two phenomena show that policy regimes of increased treatment and increased law enforcement actions can co-exist. The oft-heard phrase "treatment versus law enforcement" may represent a false dichotomy in drug policy analysis. PMID- 19129907 TI - Formal Alder-ene reaction of a bicyclo[1.1.0]butane in the synthesis of the tricyclic quaternary ammonium core of daphniglaucins. AB - A tricyclic substructure of the tetracyclic nitrogen core of the daphniglaucins was formed by an oxidative activation of the allyl side chain of a bicyclo[1.1.0]butylmethylamine, a spontaneous intramolecular formal Alder-ene reaction, and a selective cyclization of a triol intermediate. PMID- 19129908 TI - Modeling thalamocortical cell: impact of ca channel distribution and cell geometry on firing pattern. AB - The influence of calcium channel distribution and geometry of the thalamocortical cell upon its tonic firing and the low threshold spike (LTS) generation was studied in a 3-compartment model, which represents soma, proximal and distal dendrites as well as in multi-compartment model using the morphology of a real reconstructed neuron. Using an uniform distribution of Ca(2+) channels, we determined the minimal number of low threshold voltage-activated calcium channels and their permeability required for the onset of LTS in response to a hyperpolarizing current pulse. In the 3-compartment model, we found that the channel distribution influences the firing pattern only in the range of 3% below the threshold value of total T-channel density. In the multi-compartmental model, the LTS could be generated by only 64% of unequally distributed T-channels compared to the minimal number of equally distributed T-channels. For a given channel density and injected current, the tonic firing frequency was found to be inversely proportional to the size of the cell. However, when the Ca(2+) channel density was elevated in soma or proximal dendrites, then the amplitude of LTS response and burst spike frequencies were determined by the ratio of total to threshold number of T-channels in the cell for a specific geometry. PMID- 19129909 TI - Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells. AB - Astrocytes play an important role in cell-cell signaling in the mammalian central nervous system. The ability of astrocytes to communicate with surrounding cells through gap-junctional coupling or signaling via the release of transmitters makes characterization of these cells difficult in vitro and even more so in vivo. To simplify the complexity of common in vitro systems, introduced by intercellular communication between astrocytes, we developed a novel cell culturing method, in which purified rat visual cortical astrocytes were grown in spatially defined cell-adhesion wells which we termed micropits. We showed that astrocytes cultured in micropit regions were viable and exhibited similar characteristics of Ca(2+) dynamics and astrocytic marker expression to those of cells cultured in non-micropit regions. Examination of intracellular Ca(2+) oscillations in solitary astrocytes cultured in micropits revealed less variable oscillations than those of non-micropit grouped astrocytes, which were in contact with their neighbors. Solitary cells in micropit regions can undergo ATP-mediated astrocyte-microglia signaling, demonstrating that this culturing method can also be used to investigate glial-glial interactions in a spatially well-defined microenvironment. PMID- 19129910 TI - Rapid effects of marine reserves via larval dispersal. AB - Marine reserves have been advocated worldwide as conservation and fishery management tools. It is argued that they can protect ecosystems and also benefit fisheries via density-dependent spillover of adults and enhanced larval dispersal into fishing areas. However, while evidence has shown that marine reserves can meet conservation targets, their effects on fisheries are less understood. In particular, the basic question of if and over what temporal and spatial scales reserves can benefit fished populations via larval dispersal remains unanswered. We tested predictions of a larval transport model for a marine reserve network in the Gulf of California, Mexico, via field oceanography and repeated density counts of recently settled juvenile commercial mollusks before and after reserve establishment. We show that local retention of larvae within a reserve network can take place with enhanced, but spatially-explicit, recruitment to local fisheries. Enhancement occurred rapidly (2 yrs), with up to a three-fold increase in density of juveniles found in fished areas at the downstream edge of the reserve network, but other fishing areas within the network were unaffected. These findings were consistent with our model predictions. Our findings underscore the potential benefits of protecting larval sources and show that enhancement in recruitment can be manifested rapidly. However, benefits can be markedly variable within a local seascape. Hence, effects of marine reserve networks, positive or negative, may be overlooked when only focusing on overall responses and not considering finer spatially-explicit responses within a reserve network and its adjacent fishing grounds. Our results therefore call for future research on marine reserves that addresses this variability in order to help frame appropriate scenarios for the spatial management scales of interest. PMID- 19129912 TI - Adaptation of the spore discharge mechanism in the basidiomycota. AB - BACKGROUND: Spore discharge in the majority of the 30,000 described species of Basidiomycota is powered by the rapid motion of a fluid droplet, called Buller's drop, over the spore surface. In basidiomycete yeasts, and phytopathogenic rusts and smuts, spores are discharged directly into the airflow around the fungal colony. Maximum discharge distances of 1-2 mm have been reported for these fungi. In mushroom-forming species, however, spores are propelled over much shorter ranges. In gilled mushrooms, for example, discharge distances of <0.1 mm ensure that spores do not collide with opposing gill surfaces. The way in which the range of the mechanism is controlled has not been studied previously. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we report high-speed video analysis of spore discharge in selected basidiomycetes ranging from yeasts to wood-decay fungi with poroid fruiting bodies. Analysis of these video data and mathematical modeling show that discharge distance is determined by both spore size and the size of the Buller's drop. Furthermore, because the size of Buller's drop is controlled by spore shape, these experiments suggest that seemingly minor changes in spore morphology exert major effects upon discharge distance. CONCLUSIONS/SIGNIFICANCE: This biomechanical analysis of spore discharge mechanisms in mushroom-forming fungi and their relatives is the first of its kind and provides a novel view of the incredible variety of spore morphology that has been catalogued by traditional taxonomists for more than 200 years. Rather than representing non-selected variations in micromorphology, the new experiments show that changes in spore architecture have adaptive significance because they control the distance that the spores are shot through air. For this reason, evolutionary modifications to fruiting body architecture, including changes in gill separation and tube diameter in mushrooms, must be tightly linked to alterations in spore morphology. PMID- 19129911 TI - Reproductive intentions and outcomes among women on antiretroviral therapy in rural Uganda: a prospective cohort study. AB - BACKGROUND: Antiretroviral therapy (ART) may influence the biological, social and behavioral determinants of pregnancy in HIV-infected women. However, there are limited longitudinal data on the reproductive intentions and outcomes among women on ART in Africa. METHODOLOGY /PRINCIPAL FINDINGS: Using a prospective cohort design, we analyzed trends in desire for children and predictors of pregnancy among a cohort of 733 HIV-infected women in rural Uganda who initiated ART between May 2003 and May 2004 and were followed up in their homes until June 2006. Women answered in-depth social and behavioral questionnaires administered every quarter in year 1 after initiating ART, and every 6 to 12 months thereafter. Use of family planning methods was assessed at 18 and 24 months after starting ART. We tested for non-constant pregnancy incidence by using a shape parameter test from the Weibull distribution. We modeled repeated measurements of all variables related to the women's desire for children over time using a generalized estimating equation (GEE) extension to the logistic regression model. Risk factors for pregnancy were examined using Cox proportional hazards model. 711 women eligible for the study were followed-up for a median time of 2.4 years after starting ART. During this time, less than 7% of women reported wanting more children at any time point yet 120 (16.9%) women experienced 140 pregnancies and pregnancy incidence increased from 3.46 per 100 women-years (WY) in the first quarter to 9.5 per 100 WY at 24 months (p<0.0001). This was paralleled by an increase in the proportion of women reporting sexual activity in the past 3 months, from 24.4% at baseline to 32.5% over 24 months of follow-up (p = 0.001). Only 14% of women used permanent or semi-permanent family planning methods by their second year on ART. In the multivariate model, younger age (HR = 2.71 per 10-year decrease, 95% CI: 2.95-3.78), having a BMI>18.5 (HR = 1.09, CI: 1.01 1.18) and not having used condoms consistently in the last 3 months (HR = 1.79, CI: 1.02-3.13) were independently associated with pregnancy. CONCLUSION/SIGNIFICANCE: Women on ART and their partners should be consistently counseled on the effects of ART in restoring fertility, and offered regularly free and comprehensive family planning services as part of their standard package of care. PMID- 19129913 TI - The intracellular localization of ID2 expression has a predictive value in non small cell lung cancer. AB - BACKGROUND: ID2 is a member of a subclass of transcription regulators belonging to the general bHLH (basic-helix-loop-helix) family of transcription factors. In normal cells, ID2 is responsible for regulating the balance between proliferation and differentiation. More recent studies have demonstrated that ID2 is involved in tumor progression in several cancer types such as prostate or breast. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we investigated, for the first time, the relationship between the expression of ID2 in non-small cell lung cancer (NSCLC) patients and the clinicopathological features and prognosis of these patients. Immunohistochemistry was performed on tissue microarrays, which included 62 NSCLC tumors. In malignant tissues, ID2 expression has been detected in both the nuclear and cytoplasmic compartments, but we have demonstrated that only nuclear expression of ID2 is inversely correlated with the differentiation grade of the tumor (p = 0.007). Interestingly, among patients with poorly differentiated tumors, high nuclear expression of ID2 was an independent and unfavorable prognostic factor for survival (p = 0.036). CONCLUSIONS: These results suggest that ID2 could be involved in tumor dedifferentiation processes of NSCLC, and could be used as prognostic marker for patients with poorly differentiated tumors. PMID- 19129914 TI - Loss of ribosomal protein L11 affects zebrafish embryonic development through a p53-dependent apoptotic response. AB - Ribosome is responsible for protein synthesis in all organisms and ribosomal proteins (RPs) play important roles in the formation of a functional ribosome. L11 was recently shown to regulate p53 activity through a direct binding with MDM2 and abrogating the MDM2-induced p53 degradation in response to ribosomal stress. However, the studies were performed in cell lines and the significance of this tumor suppressor function of L11 has yet to be explored in animal models. To investigate the effects of the deletion of L11 and its physiological relevance to p53 activity, we knocked down the rpl11 gene in zebrafish and analyzed the p53 response. Contrary to the cell line-based results, our data indicate that an L11 deficiency in a model organism activates the p53 pathway. The L11-deficient embryos (morphants) displayed developmental abnormalities primarily in the brain, leading to embryonic lethality within 6-7 days post fertilization. Extensive apoptosis was observed in the head region of the morphants, thus correlating the morphological defects with apparent cell death. A decrease in total abundance of genes involved in neural patterning of the brain was observed in the morphants, suggesting a reduction in neural progenitor cells. Upregulation of the genes involved in the p53 pathway were observed in the morphants. Simultaneous knockdown of the p53 gene rescued the developmental defects and apoptosis in the morphants. These results suggest that ribosomal dysfunction due to the loss of L11 activates a p53-dependent checkpoint response to prevent improper embryonic development. PMID- 19129915 TI - Molecular time-course and the metabolic basis of entry into dauer in Caenorhabditis elegans. AB - When Caenorhabditis elegans senses dauer pheromone (daumone), signaling inadequate growth conditions, it enters the dauer state, which is capable of long term survival. However, the molecular pathway of dauer entry in C. elegans has remained elusive. To systematically monitor changes in gene expression in dauer paths, we used a DNA microarray containing 22,625 gene probes corresponding to 22,150 unique genes from C. elegans. We employed two different paths: direct exposure to daumone (Path 1) and normal growth media plus liquid culture (Path 2). Our data reveal that entry into dauer is accomplished through the multi-step process, which appears to be compartmentalized in time and according to metabolic flux. That is, a time-course of dauer entry in Path 1 shows that dauer larvae formation begins at post-embryonic stage S4 (48 h) and is complete at S6 (72 h). Our results also suggest the presence of a unique adaptive metabolic control mechanism that requires both stage-specific expression of specific genes and tight regulation of different modes of fuel metabolite utilization to sustain the energy balance in the context of prolonged survival under adverse growth conditions. It is apparent that worms entering dauer stage may rely heavily on carbohydrate-based energy reserves, whereas dauer larvae utilize fat or glyoxylate cycle-based energy sources. We created a comprehensive web-based dauer metabolic database for C. elegans (www.DauerDB.org) that makes it possible to search any gene and compare its relative expression at a specific stage, or evaluate overall patterns of gene expression in both paths. This database can be accessed by the research community and could be widely applicable to other related nematodes as a molecular atlas. PMID- 19129916 TI - Autophagy and exosomes in the aged retinal pigment epithelium: possible relevance to drusen formation and age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a major cause of loss of central vision in the elderly. The formation of drusen, an extracellular, amorphous deposit of material on Bruch's membrane in the macula of the retina, occurs early in the course of the disease. Although some of the molecular components of drusen are known, there is no understanding of the cell biology that leads to the formation of drusen. We have previously demonstrated increased mitochondrial DNA (mtDNA) damage and decreased DNA repair enzyme capabilities in the rodent RPE/choroid with age. In this study, we found that drusen in AMD donor eyes contain markers for autophagy and exosomes. Furthermore, these markers are also found in the region of Bruch's membrane in old mice. By in vitro modeling increased mtDNA damage induced by rotenone, an inhibitor of mitochondrial complex I, in the RPE, we found that the phagocytic activity was not altered but that there were: 1) increased autophagic markers, 2) decreased lysosomal activity, 3) increased exocytotic activity and 4) release of chemoattractants. Exosomes released by the stressed RPE are coated with complement and can bind complement factor H, mutations of which are associated with AMD. We speculate that increased autophagy and the release of intracellular proteins via exosomes by the aged RPE may contribute to the formation of drusen. Molecular and cellular changes in the old RPE may underlie susceptibility to genetic mutations that are found in AMD patients and may be associated with the pathogenesis of AMD in the elderly. PMID- 19129917 TI - A trouble shared is a trouble halved: social context and status affect pain in mouse dyads. AB - In mice behavioral response to pain is modulated by social status. Recently, social context also has been shown to affect pain sensitivity. In our study, we aimed to investigate the effects of interaction between status and social context in dyads of outbred CD-1 male mice in which the dominance/submission relationship was stable. Mice were assessed for pain response in a formalin (1% concentration) test either alone (individually tested-IT), or in pairs of dominant and subordinate mice. In the latter condition, they could be either both injected (BI) or only one injected (OI) with formalin. We observed a remarkable influence of social context on behavioral response to painful stimuli regardless of the social status of the mice. In the absence of differences between OI and IT conditions, BI mice exhibited half as much Paw-licking behavior than OI group. As expected, subordinates were hypoalgesic in response to the early phase of the formalin effects compared to dominants. Clear cut-differences in coping strategies of dominants and subordinates appeared. The former were more active, whereas the latter were more passive. Finally, analysis of behavior of the non injected subjects (the observers) in the OI dyads revealed that dominant observers were more often involved in Self-grooming behavior upon observation of their subordinate partner in pain. This was not the case for subordinate mice observing the pain response of their dominant partner. In contrast, subordinate observers Stared at the dominant significantly more frequently compared to observer dominants in other dyads. The observation of a cagemate in pain significantly affected the observer's behavior. Additionally, the quality of observer's response was also modulated by the dominance/submission relationship. PMID- 19129918 TI - Synergistic apoptosis induction in leukemic cells by the phosphatase inhibitor salubrinal and proteasome inhibitors. AB - BACKGROUND: Cells adapt to endoplasmic reticulum (ER)-stress by arresting global protein synthesis while simultaneously activating specific transcription factors and their downstream targets. These processes are mediated in part by the phosphorylation-dependent inactivation of the translation initiation factor eIF2alpha. Following restoration of homeostasis protein synthesis is resumed when the serine/threonine-protein phosphatase PP1 dephosphorylates and reactivates eIF2alpha. Proteasome inhibitors, used to treat multiple myeloma patients evoke ER-stress and apoptosis by blocking the ER-associated degradation of misfolded proteins (ERAD), however, the role of eIF2alpha phosphorylation in leukemic cells under conditions of proteasome inhibitor-mediated ER stress is currently unclear. METHODOLOGY AND PRINCIPAL FINDINGS: Bcr-Abl-positive and negative leukemic cell lines were used to investigate the functional implications of PP1-related phosphatase activities on eIF2alpha phosphorylation in proteasome inhibitor mediated ER stress and apoptosis. Rather unexpectedly, salubrinal, a recently identified PP1 inhibitor capable to protect against ER stress in various model systems, strongly synergized with proteasome inhibitors to augment apoptotic death of different leukemic cell lines. Salubrinal treatment did not affect the phosphorlyation status of eIF2alpha. Furthermore, the proapoptotic effect of salubrinal occurred independently from the chemical nature of the proteasome inhibitor, was recapitulated by a second unrelated phosphatase inhibitor and was unaffected by overexpression of a dominant negative eIF2alpha S51A variant that can not be phosphorylated. Salubrinal further aggravated ER-stress and proteotoxicity inflicted by the proteasome inhibitors on the leukemic cells since characteristic ER stress responses, such as ATF4 and CHOP synthesis, XBP1 splicing, activation of MAP kinases and eventually apoptosis were efficiently abrogated by the translational inhibitor cycloheximide. CONCLUSIONS: Although PP1 activity does not play a major role in regulating the ER stress response in leukemic cells, phosphatase signaling nevertheless significantly limits proteasome inhibitor-mediated ER-stress and apoptosis. Inclusion of specific phosphatase inhibitors might therefore represent an option to improve current proteasome inhibitor-based treatment modalities for hematological cancers. PMID- 19129920 TI - Response to Intervention with Older Students with Reading Difficulties. AB - Addressing the literacy needs of secondary school students involves efforts to raise the achievement levels of all students and to address specifically the needs of struggling readers. One approach to this problem is to consider the application of a Response to Intervention (RTI) model with older students. We describe an approach to enhanced literacy instruction for middle school students that includes the essential components of any RTI model: universal screening, progress monitoring, and multi-tiered instructional service delivery. We use screening and progress-monitoring tools specifically tied to state accountability tests and a multi-tiered instructional framework that addresses the literacy needs of all middle school students, including struggling readers. Presently a large-scale, multi-site randomized trial is under way to evaluate the feasibility and effectiveness of this RTI model for middle school students. PMID- 19129919 TI - Resurrection of a bull by cloning from organs frozen without cryoprotectant in a 80 degrees c freezer for a decade. AB - Frozen animal tissues without cryoprotectant have been thought to be inappropriate for use as a nuclear donor for somatic cell nuclear transfer (SCNT). We report the cloning of a bull using cells retrieved from testicles that had been taken from a dead animal and frozen without cryoprotectant in a -80 degrees C freezer for 10 years. We obtained live cells from defrosted pieces of the spermatic cords of frozen testicles. The cells proliferated actively in culture and were apparently normal. We transferred 16 SCNT embryos from these cells into 16 synchronized recipient animals. We obtained five pregnancies and four cloned calves developed to term. Our results indicate that complete genome sets are maintained in mammalian organs even after long-term frozen-storage without cryoprotectant, and that live clones can be produced from the recovered cells. PMID- 19129921 TI - Cell Interactome: Good Neighbors or Bad Neighbors. AB - We report an interesting finding that cellular distribution and interactions between neighboring cells modulate protein expression and localization. MCF7 breast cancer cells, which express ERb, were cultured in estrogen free medium. ERb exhibited a unique migration pattern that was dependent on cell density and proximity. This findings attest to a novel mechanism other than ligand-receptor one. Since cells were grown in conditioned culture media, the only factor influencing protein distribution is cell-cell proximity (cell-cell interactions). Thus, hormonal receptors may function independently from their ligand. in addition, physical intercellular interactions may function as a non-molecular factor inducing gene expression and activation. PMID- 19129922 TI - Out of the shadows: CXC chemokines in promoting aberrant lung cancer angiogenesis. PMID- 19129923 TI - SLM Microscopy: Scanless Two-Photon Imaging and Photostimulation with Spatial Light Modulators. AB - Laser microscopy has generally poor temporal resolution, caused by the serial scanning of each pixel. This is a significant problem for imaging or optically manipulating neural circuits, since neuronal activity is fast. To help surmount this limitation, we have developed a "scanless" microscope that does not contain mechanically moving parts. This microscope uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern. This allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision. To demonstrate the usefulness of this microscope, we perform two-photon uncaging of glutamate to activate dendritic spines and cortical neurons in brain slices. We also use it to carry out fast (60 Hz) two-photon calcium imaging of action potentials in neuronal populations. Thus, SLM microscopy appears to be a powerful tool for imaging and optically manipulating neurons and neuronal circuits. Moreover, the use of SLMs expands the flexibility of laser microscopy, as it can substitute traditional simple fixed lenses with any calculated lens function. PMID- 19129924 TI - PyMOOSE: Interoperable Scripting in Python for MOOSE. AB - Python is emerging as a common scripting language for simulators. This opens up many possibilities for interoperability in the form of analysis, interfaces, and communications between simulators. We report the integration of Python scripting with the Multi-scale Object Oriented Simulation Environment (MOOSE). MOOSE is a general-purpose simulation system for compartmental neuronal models and for models of signaling pathways based on chemical kinetics. We show how the Python scripting version of MOOSE, PyMOOSE, combines the power of a compiled simulator with the versatility and ease of use of Python. We illustrate this by using Python numerical libraries to analyze MOOSE output online, and by developing a GUI in Python/Qt for a MOOSE simulation. Finally, we build and run a composite neuronal/signaling model that uses both the NEURON and MOOSE numerical engines, and Python as a bridge between the two. Thus PyMOOSE has a high degree of interoperability with analysis routines, with graphical toolkits, and with other simulators. PMID- 19129925 TI - The Demographic Faces of the Elderly. AB - Much of the world is aging rapidly. Both the number and proportion of people aged 65 years and older are increasing, although at different rates in different parts of the world. The number of older adults has risen more than threefold since 1950, from approximately 130 million to 419 million in 2000, with the elderly share of the population increasing from 4 percent to 7 percent during that period. In the United States, those aged 65 and older currently make up about 13 percent of the population. The US Census Bureau (2004) projects that in 25 years this proportion will exceed 20 percent. Over the next 50 years the United States will undergo a profound transformation, becoming a mature nation in which one citizen in five is 65 or older. (Now, one person in eight is that old.) The dramatic increases to come in the older population will exert powerful pressures on health care delivery systems, on programs such as Social Security, Medicare, Medicaid, and Supplemental Security Income that provide financial support, and on social institutions such as the family that provide instrumental, financial, and emotional support for the elderly. PMID- 19129926 TI - Community-based free prostate cancer screening program. AB - BACKGROUND: The objective of this study was to demonstrate both the feasibility and importance of a bilingual community-based educational program that offers free prostate cancer screening to Hispanic and black men in a high-risk community. METHODS: The East Harlem Partnership for Cancer Awareness (EHPCA) brought together an academic medical center, public hospital, and two community health centers to offer monthly free prostate cancer screening programs in neighborhood community health centers. The programs included an educational component in Spanish and English, digital rectal examinations (DRE), and blood testing for prostate-specific antigen (PSA). RESULTS: Participant evaluations of the program demonstrated that the sessions were informative and helpful as well as successful in increasing knowledge about prostate cancer. Almost 90% of the participants chose to undergo a digital examination and PSA test; 4 of 150 participants were diagnosed as having prostate cancer. CONCLUSION: Offering free prostate screening is a successful method of reaching men who might otherwise not be tested. We found that men were willing to be tested and follow-up with care, as needed. PMID- 19129927 TI - A phase I/II study of a MUC1 peptide pulsed autologous dendritic cell vaccine as adjuvant therapy in patients with resected pancreatic and biliary tumors. AB - Pancreatic and biliary cancers are relatively resistant to chemotherapy and radiation and may therefore provide an opportunity for testing the potential of immunotherapy. MUC1 is an epithelial cell glycoprotein that is highly overexpressed and aberrantly glycosylated in many adenocarcinomas, including pancreatic tumors, providing a tumor specific antigen and target. We performed a Phase I/II clinical trial of a MUC1 peptide-loaded DC vaccine in 12 pancreatic and biliary cancer patients following resection of their primary tumors. The primary endpoints were vaccine toxicity and immunogenicity and the secondary endpoint was clinical outcome. The vaccine was well tolerated and no toxicity was observed. Three patients had pre-existing MUC1 antibody responses that remained stable post vaccination. MUC1-specific T cell responses were difficult to evaluate due to increases in activity of all CD8 and CD4 T cells following each vaccination. Prior to vaccination, patients entered onto this trial had a significantly higher percentage of FoxP3+CD4+ T cells compared to age matched healthy controls. The percentage of these cells also increased transiently following each injection, returning to baseline or below before the next injection. Vaccinated patients have been followed for over four years and four of the twelve patients are alive, all without evidence of recurrence. Study of the immune parameters in long-term survivors several years after vaccination may yield the sought after immune correlates of clinical responses that analysis of immune responses shortly after vaccination has not revealed. PMID- 19129928 TI - BrainMaps.org - Interactive High-Resolution Digital Brain Atlases and Virtual Microscopy. AB - BrainMaps.org is an interactive high-resolution digital brain atlas and virtual microscope that is based on over 20 million megapixels of scanned images of serial sections of both primate and non-primate brains and that is integrated with a high-speed database for querying and retrieving data about brain structure and function over the internet. Complete brain datasets for various species, including Homo sapiens, Macaca mulatta, Chlorocebus aethiops, Felis catus, Mus musculus, Rattus norvegicus, and Tyto alba, are accessible online. The methods and tools we describe are useful for both research and teaching, and can be replicated by labs seeking to increase accessibility and sharing of neuroanatomical data. These tools offer the possibility of visualizing and exploring completely digitized sections of brains at a sub-neuronal level, and can facilitate large-scale connectional tracing, histochemical and stereological analyses. PMID- 19129929 TI - THE INCIDENCE OF SUDDEN CARDIAC DEATH IN AUSTRIA. AB - The aim of the study was to assess the time structure (chronome) of sudden cardiac death (SCD) in Austria. The daily incidence of SCD (ICD-10 I46.1) in Austria was obtained for the 4-year span from Jan 2002 to Dec 2005. Data were available separately for men and women. This data series was analyzed by linear nonlinear rhythmometry. The major feature is the detection of a cis-half-year that is validated nonlinearly, the estimated period of the cis-half-year is 0.408 year (95% CI: 0.389, 0.426). It is concluded that the chronobiological analysis of sudden cardiac death in Austria showed the variability of total incidence with the period of a cis-half-year. PMID- 19129930 TI - The Benefits from Marriage and Religion in the United States: A Comparative Analysis. AB - America is a religious nation. The vast majority of Americans, when asked, profess a belief in God and affirm that religion is at least "fairly important" in their lives (Myers 2000: 285); about 60 percent of the population report membership in a religious organization and 45 percent state that they attend religious services at least monthly (Sherkat and Ellison 1999). Most American adults are currently married and almost all will marry at some time in their lives. About two-thirds of children live with their married (biological or adoptive) parents ( U.S. Census Bureau 2001). And marriage and a happy family life are almost universal goals for young adults. PMID- 19129931 TI - Trajectory to a first episode of psychosis: a qualitative research study with families. AB - AIM: The trajectory in psychotic disorders which leads from a relatively normal premorbid state in young people to a first episode of psychosis is only partly understood. Qualitative research methods can be used to begin to elucidate the temporal unfolding of symptoms leading to a first episode of psychosis, and its impact on families. METHODS: We conducted open-ended interviews with family members of 13 patients with recent onset non-affective psychotic disorders, which focused on changes observed, effects on the family, explanatory models, help seeking patterns and future expectations. Standard data analytic methods employed for qualitative research were used. RESULTS: Narratives by family members were remarkably similar. First, social withdrawal and mood symptoms developed in previously normal children; these changes were typically ascribed to drugs or stress, or to the 'storminess' of adolescence. Coping strategies by family members included prayer and reasoning/persuasion with the young person, and family initially sought help from friends and religious leaders. Entry into the mental health system was then catalysed by the emergence of overt symptoms, such as 'hearing voices', or violent or bizarre behaviour. Family members perceived inpatient hospitalization as traumatic or difficult, and had diminished expectations for the future. CONCLUSIONS: Understanding families' explanatory models for symptoms and behavioural changes, and their related patterns of help seeking, may be useful for understanding evolution of psychosis and for the design of early intervention programmes. Dissatisfaction with hospitalization supports the mandate to improve systems of care for recent-onset psychosis patients, including destigmatization and a focus on recovery. PMID- 19129932 TI - Domain-Specific and Domain-General Learning Factors are Expressed in Genetically Heterogeneous CD-1 mice. AB - It has been established that both domain-specific (e.g. spatial) as well as domain-general (general intelligence) factors influence human cognition. However, the separation of these processes has rarely been attempted in studies using laboratory animals. Previously, we have found that the performances of outbred mice across a wide range of learning tasks correlate in such a way that a single factor can explain 30- 44% of the variance between animals. This general learning factor is in some ways qualitatively and quantitatively analogous to general intelligence in humans. The complete structure of cognition in mice, however, has not been explored due to the limited sample sizes of our previous analyses. Here we report a combined analysis from 241 CD-1 mice tested in five primary learning tasks, and a subset of mice tested in two additional learning tasks. At least two (possibly three) of the seven learning tasks placed explicit demands on spatial and/or hippocampus-dependent processing abilities. Consistent with previous findings, we report a robust general factor influencing learning in mice that accounted for 38% of the variance across tasks. In addition, a domain-specific factor was found to account for performance on that subset of tasks that shared a dependence on hippocampal and/or spatial processing. These results provide further evidence for a general learning/cognitive factor in genetically heterogeneous mice. Furthermore (and similar to human cognitive performance), these results suggest a hierarchical structure to cognitive processes in this genetically heterogeneous species. PMID- 19129933 TI - Quantitative Orientation-Independent Differential Interference Contrast (DIC) Microscopy Coupled with Orientation-Independent Polarization Microscopy. PMID- 19129935 TI - Analyses of Markov decision process structure regarding the possible strategic use of interacting memory systems. AB - Behavioral tasks are often used to study the different memory systems present in humans and animals. Such tasks are usually designed to isolate and measure some aspect of a single memory system. However, it is not necessarily clear that any given task actually does isolate a system or that the strategy used by a subject in the experiment is the one desired by the experimenter. We have previously shown that when tasks are written mathematically as a form of partially observable Markov decision processes, the structure of the tasks provide information regarding the possible utility of certain memory systems. These previous analyses dealt with the disambiguation problem: given a specific ambiguous observation of the environment, is there information provided by a given memory strategy that can disambiguate that observation to allow a correct decision? Here we extend this approach to cases where multiple memory systems can be strategically combined in different ways. Specifically, we analyze the disambiguation arising from three ways by which episodic-like memory retrieval might be cued (by another episodic-like memory, by a semantic association, or by working memory for some earlier observation). We also consider the disambiguation arising from holding earlier working memories, episodic-like memories or semantic associations in working memory. From these analyses we can begin to develop a quantitative hierarchy among memory systems in which stimulus-response memories and semantic associations provide no disambiguation while the episodic memory system provides the most flexible disambiguation, with working memory at an intermediate level. PMID- 19129936 TI - Structural plasticity controlled by calcium based correlation detection. helias@bccn.uni-freiburg.de. AB - Hebbian learning in cortical networks during development and adulthood relies on the presence of a mechanism to detect correlation between the presynaptic and the postsynaptic spiking activity. Recently, the calcium concentration in spines was experimentally shown to be a correlation sensitive signal with the necessary properties: it is confined to the spine volume, it depends on the relative timing of pre- and postsynaptic action potentials, and it is independent of the spine's location along the dendrite. NMDA receptors are a candidate mediator for the correlation dependent calcium signal. Here, we present a quantitative model of correlation detection in synapses based on the calcium influx through NMDA receptors under realistic conditions of irregular pre- and postsynaptic spiking activity with pairwise correlation. Our analytical framework captures the interaction of the learning rule and the correlation dynamics of the neurons. We find that a simple thresholding mechanism can act as a sensitive and reliable correlation detector at physiological firing rates. Furthermore, the mechanism is sensitive to correlation among afferent synapses by cooperation and competition. In our model this mechanism controls synapse formation and elimination. We explain how synapse elimination leads to firing rate homeostasis and show that the connectivity structure is shaped by the correlations between neighboring inputs. PMID- 19129937 TI - Calcium, synaptic plasticity and intrinsic homeostasis in purkinje neuron models. AB - We recently reproduced the complex electrical activity of a Purkinje cell (PC) with very different combinations of ionic channel maximum conductances, suggesting that a large parameter space is available to homeostatic mechanisms. It has been hypothesized that cytoplasmic calcium concentrations control the homeostatic activity sensors. This raises many questions for PCs since in these neurons calcium plays an important role in the induction of synaptic plasticity. To address this question, we generated 148 new PC models. In these models the somatic membrane voltages are stable, but the somatic calcium dynamics are very variable, in agreement with experimental results. Conversely, the calcium signal in spiny dendrites shows only small variability. We demonstrate that this localized control of calcium conductances preserves the induction of long-term depression for all models. We conclude that calcium is unlikely to be the sole activity-sensor in this cell but that there is a strong relationship between activity homeostasis and synaptic plasticity. PMID- 19129938 TI - Local-Circuit Phenotypes of Layer 5 Neurons in Motor-Frontal Cortex of YFP-H Mice. AB - Layer 5 pyramidal neurons comprise an important but heterogeneous group of cortical projection neurons. In motor-frontal cortex, these neurons are centrally involved in the cortical control of movement. Recent studies indicate that local excitatory networks in mouse motor-frontal cortex are dominated by descending pathways from layer 2/3 to 5. However, those pathways were identified in experiments involving unlabeled neurons in wild type mice. Here, to explore the possibility of class-specific connectivity in this descending pathway, we mapped the local sources of excitatory synaptic input to a genetically labeled population of cortical neurons: YFP-positive layer 5 neurons of YFP-H mice. We found, first, that in motor cortex, YFP-positive neurons were distributed in a double blade, consistent with the idea of layer 5B having greater thickness in frontal neocortex. Second, whereas unlabeled neurons in upper layer 5 received their strongest inputs from layer 2, YFP-positive neurons in the upper blade received prominent layer 3 inputs. Third, YFP-positive neurons exhibited distinct electrophysiological properties, including low spike frequency adaptation, as reported previously. Our results with this genetically labeled neuronal population indicate the presence of distinct local-circuit phenotypes among layer 5 pyramidal neurons in mouse motor-frontal cortex, and present a paradigm for investigating local circuit organization in other genetically labeled populations of cortical neurons. PMID- 19129939 TI - Category specificity in early perception: face and word n170 responses differ in both lateralization and habituation properties. AB - N170 event-related potential (ERP) responses to both faces and visual words raises questions about category specific processing mechanisms during early perception and their neural basis. Topographic differences across word and face N170s suggests a form of category specific processing in early perception - the word N170 is consistently left-lateralized, while less consistent evidence supports a right-lateralization for the face N170. Additionally, the face N170 shows a reduction in amplitude across consecutive individual faces, a form of habituation that might differ across studies thereby helping to explain inconsistencies in lateralization. This effect remains unexplored for visual words. The current study directly contrasts N170 responses to words and faces within the same subjects, examining both category-level habituation and lateralization effects. ERP responses to a series of different faces and words were collected under two contexts: blocks that alternated faces and words vs. pure blocks of a single category designed to induce category-level habituation. Global and occipito-temporal measures of N170 amplitude demonstrated an interaction between category (words, faces) and block context (alternating categories, same category). N170 amplitude demonstrated class-level habituation for faces but not words. Furthermore, the pure block context diminished the right lateralization of the face N170, pointing to class-level habituation as a factor that might drive inconsistencies in findings of right-lateralization across different paradigms. No analogous effect for the word N170 was found, suggesting category specificity for this form of habituation. Taken together, topographic and habituation effects suggest distinct forms of perceptual processing drive the face N170 and the visual word form N170. PMID- 19129940 TI - Impact and cost-effectiveness of culture for diagnosis of tuberculosis in HIV infected Brazilian adults. AB - BACKGROUND: Culture of Mycobacterium tuberculosis currently represents the closest "gold standard" for diagnosis of tuberculosis (TB), but operational data are scant on the impact and cost-effectiveness of TB culture for human immunodeficiency (HIV-) infected individuals in resource-limited settings. METHODOLOGY/PRINCIPAL FINDINGS: We recorded costs, laboratory results, and dates of initiating TB therapy in a centralized TB culture program for HIV-infected patients in Rio de Janeiro, Brazil, constructing a decision-analysis model to estimate the incremental cost-effectiveness of TB culture from the perspective of a public-sector TB control program. Of 217 TB suspects presenting between January 2006 and March 2008, 33 (15%) had culture-confirmed active tuberculosis; 23 (70%) were smear-negative. Among smear-negative, culture-positive patients, 6 (26%) began TB therapy before culture results were available, 11 (48%) began TB therapy after culture result availability, and 6 (26%) did not begin TB therapy within 180 days of presentation. The cost per negative culture was US$17.52 (solid media)-$23.50 (liquid media). Per 1,000 TB suspects and compared with smear alone, TB culture with solid media would avert an estimated eight TB deaths (95% simulation interval [SI]: 4, 15) and 37 disability-adjusted life years (DALYs) (95% SI: 13, 76), at a cost of $36 (95% SI: $25, $50) per TB suspect or $962 (95% SI: $469, $2642) per DALY averted. Replacing solid media with automated liquid culture would avert one further death (95% SI: -1, 4) and eight DALYs (95% SI: 4, 23) at $2751 per DALY (95% SI: $680, dominated). The cost-effectiveness of TB culture was more sensitive to characteristics of the existing TB diagnostic system than to the accuracy or cost of TB culture. CONCLUSIONS/SIGNIFICANCE: TB culture is potentially effective and cost-effective for HIV-positive patients in resource-constrained settings. Reliable transmission of culture results to patients and integration with existing systems are essential. PMID- 19129941 TI - A novel and critical role for Oct4 as a regulator of the maternal-embryonic transition. AB - BACKGROUND: Compared to the emerging embryonic stem cell (ESC) gene network, little is known about the dynamic gene network that directs reprogramming in the early embryo. We hypothesized that Oct4, an ESC pluripotency regulator that is also highly expressed at the 1- to 2-cell stages in embryos, may be a critical regulator of the earliest gene network in the embryo. METHODOLOGY/PRINCIPAL FINDINGS: Using antisense morpholino oligonucleotide (MO)-mediated gene knockdown, we show that Oct4 is required for development prior to the blastocyst stage. Specifically, Oct4 has a novel and critical role in regulating genes that encode transcriptional and post-transcriptional regulators as early as the 2-cell stage. Our data suggest that the key function of Oct4 may be to switch the developmental program from one that is predominantly regulated by post transcriptional control to one that depends on the transcriptional network. Further, we propose to rank candidate genes quantitatively based on the inter embryo variation in their differential expression in response to Oct4 knockdown. Of over 30 genes analyzed according to this proposed paradigm, Rest and Mta2, both of which have established pluripotency functions in ESCs, were found to be the most tightly regulated by Oct4 at the 2-cell stage. CONCLUSIONS/SIGNIFICANCE: We show that the Oct4-regulated gene set at the 1- to 2-cell stages of early embryo development is large and distinct from its established network in ESCs. Further, our experimental approach can be applied to dissect the gene regulatory network of Oct4 and other pluripotency regulators to deconstruct the dynamic developmental program in the early embryo. PMID- 19129944 TI - Abstracts from the 1st Canadian Respiratory Conference, June 19-21, 2008, Montreal, Quebec, Canada. PMID- 19129945 TI - An overview of hepatocellular carcinoma study by omics-based methods. AB - Hepatocellular carcinoma (HCC) is one of the most deadly malignancies worldwide. Scientists have been studying the molecular mechanism of HCC for years, but the understanding of it remains incomplete and scattered across the literature at different molecular levels. Chromosomal aberrations, epigenetic abnormality and changes of gene expression have been reported in HCC. High-throughput omics technologies have been widely applied, aiming at the discovery of candidate biomarkers for cancer staging, prediction of recurrence and prognosis, and treatment selection. Large amounts of data on genetic and epigenetic abnormalities, gene expression profiles, microRNA expression profiles and proteomics have been accumulating, and bioinformatics is playing a more and more important role. In this paper, we review the current omics-based studies on HCC at the levels of genomics, transcriptomics and proteomics. Integrating observations from multiple aspects is an essential step toward the systematic understanding of the disease. PMID- 19129946 TI - Protein engineering, expression, and activity of a novel fusion protein possessing keratinocyte growth factor 2 and fibronectin. AB - Growth factor-induced proliferation and differentiation often require adhesion of cells to the extracellular matrix proteins such as fibronectin (FN). In this study, we aimed to investigate the effect of protein engineering of the keratinocyte growth factor 2 (KGF2) fused to the FN on the mitogenic activity of KGF2. The fusion protein (KGF2-FN10), which was expressed in Escherichia coli, showed significantly enhanced mitogenic activity of KGF2 on human keratinocytes. Moreover, KGF2-FN10 fusion protein showed significantly increased activity to differentiate keratinocytes from native KGF2. In conclusion, these results suggest that KGF2-FN10 fusion protein has certain advantages over native KGF2 and may offer a novel strategy to potentiate the therapeutic effect of KGF2. PMID- 19129942 TI - Asthma worsenings: approaches to prevention and management from the Asthma Worsenings Working Group. AB - Most asthma patients prescribed maintenance asthma therapies still experience periods of asthma worsenings characterized by daytime or night-time symptoms, or an increased need for rescue medication. In fact, these episodes are highly prevalent even in patients with well-controlled disease. Published literature suggests that asthma worsenings likely represent a window of opportunity during which patients could intervene early to prevent exacerbations or further deterioration of asthma symptoms. However, current evidence suggests that most patients fail to respond or to self-manage appropriately during these periods.To address the issue of asthma worsenings, an interdisciplinary committee of respirologists, allergists, family physicians, pharmacists and certified asthma educators from across Canada developed a practical definition of asthma worsenings and provided approaches to the prevention and management of these episodes based on current literature. To date, combination inhaled corticosteroid/long-acting beta-agonist therapy, particularly single inhaler maintenance and reliever therapy, appears to be an effective strategy for preventing asthma worsenings and exacerbations. Addressing the potential barriers to appropriate patient self-management of asthma worsenings, such as failure to adequately identify and respond to worsenings, low expectations for controlling asthma, low health literacy and poor patient-health care professional communication, are also critical to the successful prevention and management of these episodes. Finally, an interdisciplinary team approach involving patients and their families, certified asthma educators, primary care physicians, pharmacists and specialists is likely to have the greatest impact on the identification, prevention and management of asthma worsenings. PMID- 19129947 TI - Functional and structural analysis of two fibrinogen-activating enzymes isolated from the venoms of Crotalus durissus terrificus and Crotalus durissus collilineatus. AB - Fibrinogen-activating enzymes, widely distributed in Crotalidae and Viperidae venoms, are single-chain glycosylated serine proteases that display high macromolecular selectivity and are often referred to as thrombin-like enzymes (TLEs). TLEs serve as structural models to extend our understanding of the structure-function relationships of blood coagulation factors, have been clinically used for the treatment of thrombotic diseases, and are used as tools in clinical assays. The combination of gel filtration and ion-exchange chromatography proved to be successful in obtaining milligram quantities of pure samples of TLEs from the venoms of Crotalus durissus terrificus (white venom) and Crotalus durissus collilineatus (yellow venom). Functional characterization indicates that both enzymes preferentially degrade the Bb chain of bovine fibrinogen and possess edema-inducing and coagulant activities. However, the TLE from C. d. collilineatus venom shows twofold higher coagulant activity with a minimum coagulant dose (MCD) of 0.6 microg/microl, whereas the enzyme isolated from C. d. terrificus indicated an MCD of 1.5 microg/microl. Molecular modeling of gyroxin and structural comparisons with other highly conserved snake venom serine proteases, underlines the key role played by the surface charge distribution and the double insertion in the 174-surface loop in macromolecular substrate recognition by TLEs. PMID- 19129948 TI - The optional long 5'-untranslated region of human ACAT1 mRNAs impairs the production of ACAT1 protein by promoting its mRNA decay. AB - We have previously reported that human ACAT1 mRNAs produce the 50 kDa protein using the AUG(11397-1399) initiation codon, and also a minor 56 kDa isoform using the upstream in-frame GGC(1274-1276) initiation codon. The GGC(1274-1276) codon is located at the optional long 5'-untranslated region (5'-UTR, nt 1-1396) of the mRNAs. The DNA sequences corresponding to this 5'-UTR are located in two different chromosomes, 7 and 1. In the current work, we report that the optional long 5'-UTR significantly impairs the production of human ACAT1 protein initiated from the AUG(1397-1399)codon, mainly by promoting its mRNA decay. The western blot analyses indicated that the optional long 5'-UTR potently impaired the production of different proteins initiated from the AUG(1397-1399) codon, meaning that this impairing effect was not influenced by the 3'-UTR or the coding sequence of ACAT1 mRNA. The results of reverse transcription-quantitative polymerase chain reaction demonstrated that this 5'- UTR dramatically reduced the contents of its linked mRNAs. Analyses of the protein to mRNA ratios showed that this 5'-UTR mainly decreased its mRNA stability rather than altering its translational efficiency. We next performed the plasmid transfection experiments and used actinomycin D to inhibit transcription. The results showed that this 5' UTR promoted its mRNA decay. Additional transfection and nucleofection experiments using RNAs prepared in vitro illustrated that, in both the cytoplasm and the nucleus of cells, the optional long 5'-UTR-linked mRNAs decayed faster than those without the link. Overall, our study brings new insight to the regulation of the human ACAT1 gene expression at the post-transcription level. PMID- 19129949 TI - Doppel-induced cytotoxicity in human neuronal SH-SY5Y cells is antagonized by the prion protein. AB - Doppel (Dpl) is a prion (PrP)-like protein due to the structural and biochemical similarities; however, the natural functions of Dpl and PrP remain unclear. In this study, a 531-bp human PRND gene sequence encoding Dpl protein was amplified from human peripheral blood leucocytes. Full-length and various truncated human Dpl and PrP proteins were expressed and purified from Escherichia coli. Supplement of the full-length Dpl onto human neuroblastoma cell SH-SY5Y induced remarkable cytotoxicity, and the region responsible for its cytotoxicity was mapped at the middle segment of Dpl [amino acids (aa) 81-122]. Interestingly, Dpl induced cytotoxicity was antagonized by the presence of fulllength wild-type PrP. Analysis on fragments of PrP mutants showed that the N-terminal fragment (aa 23- 90) of PrP was responsible for the protective activity. A truncated PrP (PrPdelta32-121) with similar secondary structure as Dpl induced Dpl-like cytotoxicity on SHSY5Y cells. Furthermore, binding of copper ion could enhance the antagonizing effect of PrP on Dpl-induced cytotoxicity. Apoptosis assays revealed that cytotoxicity induced by Dpl occurred through an apoptotic mechanism. These results suggested that the function of Dpl is antagonistic to PrP rather than synergistic. PMID- 19129950 TI - Inactivation of LARS2, located at the commonly deleted region 3p21.3, by both epigenetic and genetic mechanisms in nasopharyngeal carcinoma. AB - Allelic loss of chromosome 3p, including the 3p21.3 region, is found in 95-100% of primary nasopharyngeal carcinoma (NPC) biopsies, suggesting that this region should harbor some tumor suppressor genes (TSGs) closely related to NPC development. Several TSGs located at 3p21.3, such as RASSF1A, LTF and BLU, have been demonstrated to be involved in NPC development. LARS2 (leucyl-tRNA synthetase 2, mitochondrial) is another gene located in the chromosome 3 common eliminated region-1 (C3CER1) at 3p21.3. In this study, we focussed on the epigenetic and genetic alterations of LARS2 in NPC. The mRNA expression of LARS2 was detected in 36 NPC and 8 chronic nasopharyngitis (NP) tissues by semi quantitative reverse transcription-polymerase chain reaction (RT-PCR) and real time RT-PCR. Subsequently, the mutation, allelic loss, and methylation status of LARS2 were analysed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), homozygous deletion (HD) analysis and methylation specific polymerase chain reaction in primary NPC tissues. No expression or downregulation of LARS2 was observed in 78% of primary NPC tissues. No mutations, assessed by PCR-SSCP and DNA sequencing, were found in the promoter region and exon 1 of LARS2 in NPC tissues, whereas HD was detected in 28% of NPC specimens at the LARS2 locus. In addition, hypermethylation of LARS2 was found in 64% of NPC samples but only in 12.5% of NP biopsies. Our data indicate that inactivation of LARS2 by both genetic and epigenetic mechanisms may be a common and important event in the carcinogenesis of NPC. PMID- 19129951 TI - PEA3 activates VEGF transcription in T47D and SKBR3 breast cancer cells. AB - Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis and a prognostic factor for many tumors, including those of endocrine-responsive tissues such as the breast and uterus. In this study, we found that overexpression of PEA3 could increase VEGF mRNA levels and VEGF promoter activity in human T47D and SKBR3 breast cancer cells. Chromatin immunoprecipitation assay demonstrated that PEA3 could bind to the VEGF promoter in the cells transfected with PEA3 expression vector. PEA3 small interfering RNA attenuated VEGF promoter activity and the binding of PEA3 to the VEGF promoter in T47D and SKBR3 cells. These results indicated that PEA3 could activate VEGF promoter transcription. PMID- 19129952 TI - Isolation and characterization of carboxypeptidase III from germinating triticale grains. AB - Carboxypeptidase III from germinating triticale grains was purified 434.2-fold with a six-step procedure including: homogenization, ammonium sulfate precipitation, cation-exchange chromatography on CM-cellulose, gel filtration chromatography on Sephadex G-150, cation-exchange chromatography on SP8HR column (HPLC), and affinity chromatography on CABSSepharose 4B. Triticale carboxypeptidase III is a monomer with a molecular weight of 45 kDa, which optimally hydrolyzes peptides at temperature 30-50 degrees C and pH 4.6. N-CBZ Ala-Phe, N-CBZ-Ala-Leu, and N-CBZ-Ala-Met are hydrolyzed at the highest rates. Amino acids with aromatic or large aliphatic side chains are preferred in position P1', whereas the presence of these types of groups in position P1 of the substrate results in a lower rate of hydrolysis. Peptides containing glutamic acid in positions P1 are poor substrates for the enzyme. This phenomenon suggests the hydrophobic substrate- binding sites S1 and S1'. The active site contains serine since diisopropylfluorophosphate and phenylmethanesulfonyl fluoride reduce the activity by 89.9% and 81.5%, respectively. Moreover, the activity of triticale carboxypeptidase III is reduced by mercury ions and organomercurial compounds, which suggests the presence of a sulfhydryl group adjacent to the active site of the enzyme. Identification of purified enzyme by mass spectrometry method demonstrated that the enzyme is a homolog of barley carboxypeptidase III. PMID- 19129953 TI - Cloning, expression, and protective immunity in mice of a gene encoding the diagnostic antigen P-29 of Echinococcus granulosus. AB - Taeniid tapeworm Echinococcus granulosus is the causative agent of Echinococcosis, an important zoonosis with worldwide distribution. In this study, a diagnostic antigen P-29 was cloned from E. granulosus and expressed in Escherichia coli. Sequence analysis showed that EgP-29 contains 717-bp open reading frame and encodes a protein of 238 amino acid residues with a predicted molecular weight of 27.1 kDa. The recombinant EgP-29 (rEgP-29) could be recognized with antimice sera in Western blotting. The specific antibody was detected by enzyme-linked immunosorbent assay. Mice vaccinated with rEgP-29 and challenged intraperitoneally with E. granulosus protoscoleces revealed significant protective immunity of 96.6% (P<0.05), compared with the control group. Thus, rEgP-29 protein is a promising candidate for an effective vaccine to prevent secondary echinococcosis. PMID- 19129954 TI - Effects of methotrexate on the developments of heart and vessel in zebrafish. AB - Methotrexate (MTX), an antagonist of folic acid, can inhibit dihydrofolate reductase (DHFR) which is of great importance in the synthesis of tetrahydrofolic acid and embryonic development. In this study, we found that after being exposed to 1.5 mM MTX at 6-10 hours post-fertilization, zebrafish embryos fail to form normal cardiovascular system. In MTX-treated embryos, the morphological development of ventricle and atrium was disrupted, the cardiac twist was abnormal, the heart rate and ventricular shortening fraction were reduced, and the vascular development was disrupted. We also found that either microinjection with dhfr-gfp mRNA or treatment with folinic acid calcium salt pentahydrate (CF) could cause improved development in the heart and vessels in MTX-treated embryos, which proved that MTX induced the malformations by inhibiting DHFR. The transcript levels of genes such as hand2, mef2a, mef2c, and flk-1 were reduced in MTXtreated embryos. Compared with the MTX-treated group, the transcript levels of hand2, mef2a, mef2c, and flk-1 were increased in the MTX 1 dhfr-gfp mRNA injected group and in the MTX 1 CF group. Our results indicated that the disrupted development of the heart and vessels in MTX-treated embryos is related to the reduced transcript levels of hand2, mef2a, mef2c, and flk-1. PMID- 19129955 TI - Food for thought ... On food safety testing. PMID- 19129956 TI - [Dignity or integrity - does the genetic modification of animals require new concepts in animal ethics?]. AB - Animal genetic engineering seems to point at a normative gap beyond pathocentric welfare theories in animal ethics. Recently developed approaches aim to bridge this gap by means of new normative criteria such as animal dignity and animal integrity. The following comparison of dignity and integrity in the context of animal ethics shows that the dignity concept faces serious problems because of its necessarily anthroporelational character and the different functions of contingent and inherent dignity within ethical reasoning. Unlike animal dignity the concept of animal integrity could prove to be a useful enhancement for pathocentric approaches. PMID- 19129957 TI - [The metaphysical dimension of animal ethics]. AB - Utilitarian ethics recognises animals as moral objects, but it does not attribute an absolute value to human or non-human individuals. Animal ethics according to Regan defines the non-human individual as an inherent value, but concedes that humans should be given precedence over animals if a situation involves a decision between life and death. Such life and death decisions relate to the fundamental structures of biological nature. To individuals these fundamental structures (the paradox of life and death) will necessarily appear absurd. The metaphysical dimension of animal ethics tries to shed light on the connections between life and death, body and mind that underly ethical discussions and searches for alternatives to the natural organisation of life. PMID- 19129958 TI - [Against the pariah's moral. Shopenhauer on the ethics of vivisection]. AB - In this essay I investigate Schopenhauer's position on the ethics of animal experiments. I argue that both his analysis of the dangers involved in performing vivisection on a wide scale and the guidelines he suggested in order to limit this cruel practice remain theoretically relevant in the context of today's debate on animal rights. Schopenhauer responded to the rapid increase in the use of animals at universities by calling for a philosophical system able to address the problems of animal ethics and treating vivisection as an important moral issue. This essay is divided into three sections: first, I reconstruct the historical background of Schopenhauer's reflections on vivisection; then I present the philosophical basis of Schopenhauer's guidelines for an ethical treatment of animals in university laboratories; finally I underline the modernity of Schopenhauer's position, focussing on a few specific examples (such as the problem of performing vivisection on apes). PMID- 19129960 TI - Predicting the outcome of shunt surgery in normal pressure hydrocephalus. PMID- 19129961 TI - Adverse radiation effects after Gamma Knife Surgery in relation to dose and volume. AB - INTRODUCTION: The relationship between target volume and adverse radiation effects (AREs) at low prescription doses requires elucidation. The development of AREs in three series of patients treated in the Gamma Knife is analysed in relation to prescription dose and target volume. MATERIALS AND METHODS: There were three groups. In group 1, there were of 275 patients with meningiomas; in group 2, 132 patients with vestibular schwannomas; and in group 3, 107 patients with arteriovenous malformations (AVMs). The minimum follow-up for each group was more than 24 months. All patients were followed up at six monthly intervals. The patients with tumours received a prescription dose of 12 Gy, which was varied to protect normal structures but not in relation to tumour volume per se. The desired AVM prescription dose was 25 Gy, but this was also reduced to protect normal structures and to keep the total dose within certain pre-defined limits. All AREs refer to intra-parenchymal increased perilesional T2 signal on MR irrespective of clinical correlation. RESULTS: There was no relationship between tumour volume and the development of ARE in the tumour groups. There was a highly significant relationship between target volume and the development of ARE for the AVMs with their much higher dose. Radiation-induced clinical trigeminal and facial nerve deficits with both vestibular schwannomas and meningiomas were always associated with an increased T2 signal in the neighbouring brainstem parenchyma. CONCLUSIONS: The relationship between target volume and the risk of adverse radiation effects may not apply with lower prescription doses. Individual radiosensitivity may explain why a minority suffer AREs unrelated to target volume. It is possible that radiation-induced brainstem parenchymal damage with concomitant cranial nerve deficits may be commoner after radiosurgery than is usually thought. If tumour control with lower doses is adequate, radiosurgery could be safely considered for larger targets associated with a high risk from microsurgery. PMID- 19129962 TI - Use of a thrombin-gelatin haemostatic matrix in endoscopic endonasal extended approaches: technical note. AB - OBJECTIVE: The management of intradural bleeding during extended endoscopic endonasal surgery is challenging. This technical note describes the use of a biological haemostatic agent which could be useful when other established strategies and materials are not effective. MATERIALS AND METHODS: From January 2004 to January 2008, 65 consecutive patients underwent extended endoscopic endonasal interventions. Of these, 29 procedures required the application of a thrombin-gelatin haemostatic matrix. We reviewed the patients' operative records to determine the source, type of bleeding and haemostatic strategy. RESULTS: We analysed bleedings from the venous sinuses, arteries, tumour bed and internal carotid artery and observed complete haemostasis immediately after application of FloSeal. The matrix was useful for both oozing and focal haemorrhage and effective even for high-flow bleeding. Only bleeding from an internal carotid artery tear required a second application. CONCLUSION: The thrombin-gelatin matrix could represent a valuable tool when other haemostatic strategies are ineffective or suboptimal. It is safe and biocompatible when compared with haemostatic agents currently in use. PMID- 19129963 TI - Risk factors for complications during intracranial electrode recording in presurgical evaluation of drug resistant partial epilepsy. AB - BACKGROUND: Intracranial electrode monitoring is still required in epilepsy surgery; however, it is associated with significant morbidity. OBJECTIVE: To identify risk factors associated with complications during invasive intracranial EEG monitoring. MATERIALS AND METHODS: Retrospective study of all patients undergoing invasive monitoring at Westmead between 1988-2004. From detailed chart reviews, the following variables were recorded: duration of intracranial monitoring, the site of grid implantation, number of grids and electrodes, seizure frequency, postoperative complications and seizure outcome. RESULTS: Seventy-one patients (median age: 24 years) underwent subdural electrode implantation; 62% had extratemporal lobe epilepsy and 46% were non-lesional. Of the 58 monitored patients who had cortical resections, 45 had good seizure outcomes. Complications related to subdural electrode implantation included transient complications requiring no treatment (12.7%), transient complications requiring treatment (9.9%) and two deaths (2.8%). Specific complications included subdural haemorrhage, transient neurological deficit, infarction and osteomyelitis. The two deaths occurred within 48 h of implantation were related to raised intracranial pressure (one venous infarction, one unexplained). Complications were associated with maximal size of grid (p < 0.001), greater number of electrodes (p < 0.001), electrode density per cortical surface implanted (p < 0.001), right central surface implantation (p = 0.003) and left central surface implantation (p = 0.013). Multiple logistic regression identified larger size grids and right central surface implantation as independent predictors of complications. CONCLUSION: There are significant complications during intracranial EEG evaluations but the majority of these are transient. We found a relationship between the size of the electrode arrays and the incidence of complications. The results of this study have been used to modify our implantation and monitoring protocols. PMID- 19129965 TI - Cross-informant correlations on social competence and behavioral problems in Sami and Norwegian preadolescents. AB - In a community-based birth cohort from Arctic Norway, correlations between parents and teachers on child competence and behavioral problems were determined for Sami and Norwegian 11-12 year-olds, using as instruments the child behavior checklist (CBCL), teacher report form (TRF), and the impact supplement of the extended strength and difficulties questionnaire (SDQ). Parent-teacher correlations on child behavioral problems were generally high in the Norwegian group, but low in the Sami group. Cross-cultural differences in cross-informant correlations were highest regarding externalizing and attention problems. Parent teacher correlations on total impact of child difficulties also differed between the ethnic groups. Once again, a lower correlation was found for the Sami children. The discrepancy between parents' and teachers' perception of problems that needed attention was highest for the Sami, and lowest for the Norwegians. The Sami parents reported fewer perceived difficulties and less impact of problems than did the Norwegian parents. In contrast, no ethnic differences emerged for teachers' ratings. The paper discusses how cultural norms might influence the reports of child problems. It demonstrates the importance of combining parent and teacher reports of child behavior problems in minority and indigenous children, who often live under different cultural norms in home and school contexts. PMID- 19129964 TI - Phenomenology, socio-demographic factors and outcome upon discharge of manic and mixed episodes in hospitalized adolescents: a chart review. AB - BACKGROUND: The existence of bipolar disorder type I (BD-I) during adolescence is now clearly established whereas there are still some controversies on BD-II and BD-NOS diagnosis, mainly in Europe (O'Dowd in Br Med J 29, 2006). Little is known on the phenomenology and potential short-term prognosis factors of bipolar episodes in this age population. In particular, very few studies examine this issue on inpatients in the European context of free access to care. OBJECTIVE: To describe the phenomenology of acute manic and mixed episodes in hospitalized adolescents and to analyse potential predictive factors associated with clinical improvement at discharge and length of hospitalization. METHODS: A total of 80 subjects, aged 12-20 years, consecutively hospitalized for a manic or mixed episode. Socio-demographic and clinical data were extracted by reviewing patients' charts. We used a multivariate analysis to evaluate short-term outcome predictors. RESULTS: The sample was characterized by severe impairment, high rates of psychotic features (N = 50, 62.5%), a long duration of stay (mean 80.4 days), and an overall good improvement (86% very much or much improved). Thirty three (41.3 %) patients had a history of depressive episodes, 13 (16.3%) had manic or brief psychotic episodes but only 3 (3.7%) had a history of attention deficit/hyperactivity disorders. More manic episodes than mixed episodes were identified in subjects with mental retardation (MR) and in subjects from migrant and/or low socio-economic families. Overall severity and female gender predicted better improvement in GAF scores. Poor insight and the existence of psychotic features predicted longer duration of stay. CONCLUSION: These results suggest that severe manic and mixed episodes in adolescents with BD-I need prolonged inpatient care to improve and that socio-cultural factors and MR should be examined more closely in youth with BD. PMID- 19129966 TI - Agreement between youth-reported and parent-reported psychopathology in a referred sample. AB - OBJECTIVE: The study examined parent-youth agreement regarding reports on psychopathology among adolescents suffering from psychiatric disorders. METHOD: A total of 1,718 patients between the age of 11 and 18, as well as their parents, were assessed using the child behavior checklist (CBCL), and the youth self report (YSR). RESULTS: Poor to low agreement between parent- and adolescent reported problem behavior on the internalizing scale, the total problem scale and moderate agreement concerning the externalizing scale of the CBCL and the YSR were found. Independent from the amount of psychiatric diagnoses, adolescents reported significantly less behavioral problems than their parents. Concerning externalizing problems, parent-youth disagreement was stronger for patients suffering from comorbid psychiatric disorders, than for adolescents displaying only one psychiatric disorder. CONCLUSION: In clinically referred children, parents are likely to emphasize the severity of the difficulties, whereas adolescents' under-report symptoms. PMID- 19129967 TI - Competitive exclusion of Cyanobacterial species in the Great Salt Lake. AB - The Great Salt Lake is separated into different salinity regimes by rail and vehicular causeways. Cyanobacterial distributions map salinity, with Aphanothece halophytica proliferating in the highly saline northern arm (27% saline), while Nodularia spumigena occurs in the less saline south (6-10%). We sought to test if cyanobacterial species abundant in the north are competitively excluded from the south, and if southern species are excluded by the high salinity of the north. Autoclaved samples from the north and south sides of each causeway were inoculated with water from each area. Aphanothece, Oscillatoria, Phormidium, and Nodularia were identified in the culture flasks using comparative differential interference contrast, fluorescence, and scanning electron microscopy. Aphanothece halophytica occurred in all inocula, but is suppressed in the presence of Nodularia spumigena. N. spumigena was found only in inocula from the less saline waters in the south, and apparently cannot survive the extremely hypersaline waters of the northern arm. These data suggest that both biotic and abiotic factors influence cyanobacterial distributions in the Great Salt Lake. PMID- 19129968 TI - Clonal diversity and geographic structure in Pleurochaete squarrosa (Pottiaceae): different sampling scale approach. AB - This paper describes our investigation of genetic variation and clonal structure of the Mediterranean moss Pleurochaete squarrosa (Brid.) Lindb. (Pottiaceae), using inter-simple sequence repeat (ISSR) molecular markers and trnL(UAA) (intron of plastid gene for Leu tRNA) sequence, choosing different sampling strategies and scales on 16 European populations. Moreover, the intercontinental distribution of two trnL haplotypes, previously found over a large area and including 24 populations in three continents, was also investigated. Despite the prevalent asexual reproduction, P. squarrosa shows a high level of genetic diversity. Some site features seem to affect the clonal structure at the local scale, influencing the relocation of detached fragments and the level of intermingling, but they do not substantially affect genetic diversity. The peculiar vegetative reproduction coupled with somatic mutation could partly account for the genetic variation detected. Genetic distances highlight geographic isolation and limited gene flow among populations. We found only two trnL haplotypes in Europe due to length polymorphism, but, over an intercontinental scale, only non-delete trnL was found in Africa and the USA. ISSR analysis within each population detected a higher genetic distance between the samples with different trnL haplotypes, suggesting the presence of two different genetic lineages within this species, geographically overlapping in the Mediterranean Basin. PMID- 19129969 TI - Ophthalmoplegic migraine in a 15-year-old Ethiopian: case report and literature review. AB - The International Headache Society (IHS) defines ophthalmoplegic migraine (OM) as recurrent attacks of headache with migrainous characteristics, associated with paresis of one or more ocular cranial nerves (commonly the third cranial nerve), and in the absence of any demonstrable intracranial lesion other than MRI changes within the affected nerve. According to the IHS criteria, it is diagnosed when at least two attacks with migraine-like headaches are accompanied with, or followed within 4 days of onset by, paresis of one or more of the third, fourth or sixth cranial nerves. Parasellar, orbital fissure and posterior fossa lesions should be ruled out by appropriate investigations. It is unlikely that OM is a variant of migraine, since the headache often lasts for a week or more and there is a latent period of up to 4 days from the onset of headache to the onset of ophthalmoplegia. Furthermore, in some cases MRI shows gadolinium uptake in the cisternal part of the affected cranial nerve and this suggests that the condition may be a recurrent demyelinating neuropathy. In general, patients demonstrated a: (1) prolonged time for symptom resolution to occur (median time 3 weeks); (2) tendency for recurrent episodes to have more severe and persistent nerve involvement; (3) evidence of permanent neurological sequelae with recurrent episodes (30% of patients); (4) rapid improvement and shortened duration with corticosteroid therapy and; (5) transient, reversible MRI contrast enhancement of the affected cranial nerve (86% of patients). Different pathogenetic mechanisms, which include compressive, ischemic and inflammatory, have been suggested for OM. Here, a 15-year-old Ethiopian with recurrent attacks of headache and third nerve palsy is presented. The subsequent discussion focuses on current evidences with regard to the clinical characteristics, possible pathogenetic mechanisms and treatment. Finally, a brief discussion of the situation in Africa will be presented. PMID- 19129970 TI - The bugs that came in from the cold: molecular adaptations to low temperatures in insects. AB - The widespread distribution of insects over many ecological niches is a testimony to their evolutionary success. The colonization of environments at high latitudes or altitudes required the evolution of biochemical strategies that reduced the impact of cold or freezing stress. This review focuses on our current interests in some of the genes and proteins involved in low temperature survival in insects. Although the most widespread form of protection is the synthesis of low molecular weight polyol cryoprotectants, proteins with intrinsic protective properties, such as the thermal hysteresis or antifreeze proteins are also important. These have been cloned and characterized in certain moths and beetles. Molecular techniques allowing the isolation of genes differentially regulated by low temperatures have revealed that heat shock proteins, cold stress proteins, membrane protectants, as well as ice nucleators and other less well characterized proteins likely also play a role in cold hardiness. PMID- 19129971 TI - A phase I study of an oral simulated FOLFOX with high dose capecitabine. AB - BACKGROUND: A phase I study of high-dose capecitabine given over 2 days, along with oxaliplatin, bolus 5FU and leucovorin (LV), was designed to simulate FOLFOX6 without the need for infusional 5FU. METHODS: Schedule A included oxaliplatin 100 mg/m(2), 5FU 400 mg/m(2), and LV 20 mg/m(2) (all given IV on days 1 and 15, 28 day cycle). Capecitabine was administered orally every 8 h x 6 doses, days 1 and 15. Schedule B excluded 5FU and LV, maintaining oxaliplatin and capecitabine. Pharmacokinetics were performed for capecitabine for 6 patients on each schedule. RESULTS: 36 patients were treated. The dose-limiting toxicities seen included nausea, dehydration, fatigue, hypotension and confusion. Minimal palmar-plantar erythrodysesthesia was seen. Myelosuppression was common, but not a dose limiting toxicity. The pharmacokinetic parameters for capecitabine were unaltered. CONCLUSION: Using capecitabine to mimic FOLFOX6 is feasible and well tolerated with a toxicity profile that differs from standard 14-day capecitabine dosing, with less palmar-plantar erythrodysesthesia. The phase II dose for capecitabine in combination with oxaliplatin, 5FU, and LV is 1,500 mg/m(2)/dose or 2,250 mg/m(2)/dose in the absence of bolus 5FU/LV. PMID- 19129973 TI - Anti-proliferative effect of the gastrin-release peptide receptor antagonist RC 3095 plus temozolomide in experimental glioblastoma models. AB - Malignant gliomas have a dismal prognosis despite multi-modality treatments like neurosurgical resection, radiation therapy and chemotherapy. Evidence has indicated that gastrin-releasing peptide (GRP) and its receptor (GRPR) play a role in the development of a variety of cancers including gliomas. In the present study, we investigated the effects of RC-3095, a selective GRPR antagonist, alone or in combination with temozolomide (TMZ), a DNA alkylating agent, in in vitro and in vivo experimental rat C6 glioma models. Cellular proliferation was significantly reduced by all treatments with the combined administration of TMZ and RC-3095 being the most effective treatment. In in vivo experiments, the control group displayed the largest tumors (52 +/- 15.5 mm(3)), whereas RC-3095 reduced the tumor size, with the most significant effect at the dose of 0.3 mg/kg (21 +/- 9.7 mm(3)). The combined therapy produced further reduction in tumor size (10 +/- 7.5 mm(3)). Our results show that the combination of RC-3095 with TMZ produced an important reduction in in vitro and in vivo glioma growth therefore making RC-3095 a candidate drug to potentiate the effects of the DNA alkylating agent TMZ in the treatment of glioma. PMID- 19129974 TI - Manual small incision cataract surgery in a United Kingdom university teaching hospital setting. AB - PURPOSE: To assess the complication rates and visual outcomes of manual small incision cataract surgery (MSICS) in a university teaching hospital setting. METHOD: Retrospective audit encompassing a 3-year period looking at the visual outcomes, and intraoperative and postoperative complications of planned MSICS performed on brunescent cataracts. RESULTS: MSICS was performed in 55 eyes of 49 patients. Intraoperatively, zonular dehiscence without vitreous loss occurred in one eye (1.8%), and vitreous loss occurred in one eye (1.8%). Postoperative complications occurred in seven eyes (12.7%). The most common complication was raised intraocular pressure (three eyes, 5.5%). None developed endophthalmitis or dropped nucleus. Mean follow-up duration was 10.0 weeks. In 49 eyes (89.1%), vision improved postoperatively. Sixty five percent achieved a best-corrected vision of 6/12 or better. The mean postoperative astigmatism was significantly higher than preoperatively (1.40D vs. 0.99D, p = 0.02). CONCLUSION: MSICS is safe and effective for dense and brunescent cataracts in a UK university hospital setting. However, it was associated with a statistically significant increase in astigmatism postoperatively. PMID- 19129975 TI - An audit of hospital based outpatient infusions and a pilot program of community based monoclonal antibody infusions. AB - INTRODUCTION: Infliximab, a chimeric monoclonal antibody to tumour necrosis factor alpha, is administered as an intravenous infusion requiring a costly hospital day case or inpatient admission. METHODS: An audit of all current therapies given by intravenous infusions in an outpatient setting in St Vincent's University Hospital (SVUH) was undertaken. Furthermore, in conjunction with TCP homecare, we established in a general practise health clinic, the first Irish community infusion centre for the administration of infliximab in August 2006. RESULTS: All outpatient departments indicated that they would favour a centralized hospital infusion unit. There were no adverse events and the mean global satisfaction improved in the community infliximab infusion pilot programme of seven patients. CONCLUSION: This study suggests efficiencies in providing centralized infusion facilities, while the community based infusion of infliximab is feasible and safe in this small cohort and identifies the community infusion unit as a viable and cost efficient alternative for administration of infliximab. PMID- 19129976 TI - Suboptimal control of blood pressure at hospital discharge in stroke patients. PMID- 19129977 TI - Potential impact of enhanced practice efficiency on endoscopy waiting times. AB - BACKGROUND: With the growing demand on endoscopy services, optimising practice efficiency has assumed increasing importance. Prior research has identified practice changes, which increase the efficiency in endoscopy. In this study, the potential impact of these practice changes on the current and projected future endoscopy waiting times at our institution was assessed. METHODS: The annual volume of endoscopic procedures performed at a major teaching hospital and the annual procedure demand from 2000 to 2007 were reviewed. Procedure demand and waiting times were projected until 2012. The impact of three practice changes, which have been shown to increase efficiency was assessed: 1. routinely obtaining i.v. access and consent in patients prior to endoscopy (approach 1); 2. routinely obtaining i.v. access and consent, and sedating the patient prior to endoscopy (approach 2); 3. utilizing a two-room per endoscopist model (approach 3). RESULTS: There has been a significant increase in annual procedure volume (36%) and annual procedure demand (69%) from 2000 to 2007. Annual waiting times for routine procedures have lengthened, from 6 weeks (2000) to 22 weeks (2007). Assuming continued linear growth in demand up to 2012, the projected waiting times will continue to rise reaching 40 weeks in 2012. Routinely obtaining i.v. access/consent prior to procedure (approach 1) would shorten the average routine waiting times so that 8 weeks (recommended HSE maximum) would not be exceeded until early 2006; routinely obtaining i.v. access/consent and sedating patient prior to procedure (approach 2) would shorten the average routine waiting time so that 8 weeks would not be exceeded until 2008; utilising two rooms per endoscopist (approach 3) would shorten the average routine waiting time so that 8 weeks would not be exceeded until early 2012. CONCLUSIONS: Maintaining timely access to endoscopic services is becoming more challenging in the face of growing demand. Modifications in routine clinical practice can significantly impact procedure waiting times. In an era where economic aspects of medical care are becoming increasingly important and where there is growing focus on waiting times as a measure of clinical performance, these findings underscore the importance of providing clinical care in the most efficient manner possible. PMID- 19129978 TI - An examination of the prevalence of IDF- and ATPIII-defined metabolic syndrome in an Irish screening population. AB - OBJECTIVES: To investigate the prevalence of ATPIII- and IDF-defined metabolic syndrome (MetS) in an Irish screening population and to determine the calculated cardiovascular risk for each group. DESIGN: A total of 1,716 subjects were enrolled over a 12-month period. MAIN OUTCOMES: The ATPIII-defined prevalence of MetS in this population was 13.2%. Using IDF criteria, 21.4% of subjects were identified as having the MetS. Correlation between the two definitions was high; however, IDF criteria identified an additional 9.5% (n = 164) of the population as having MetS, which ATPIII criteria failed to recognise. CONCLUSION: We noted a higher prevalence of MetS in the studied population when defined by IDF criteria. However, those identified by IDF and not by ATPIII definition did not have a higher cardiovascular risk score by either Framingham or European Score than those without MetS. Thus, application of the ATPIII definition of MetS, may be the more practical. PMID- 19129979 TI - Tinnitus as an unusual presentation of Schneiderian papillomatosis. AB - INTRODUCTION: Primary Schneiderian papillomatosis of the middle ear and mastoid cavity is extremely rare. It is frequently associated with intermittent unilateral otorrhoea and mass in the middle ear and mastoid cavity. METHODS: Case presentation, symptoms, diagnostic criteria, management and literature review are discussed. CONCLUSION: Schneiderian papillomatosis is an important differential diagnosis of mass in the middle ear and mastoid cavity, and tinnitus as a presenting symptom has not been reported before. Primary radical treatment is essential in preventing tumour recurrence. PMID- 19129980 TI - Importance of full evaluation in patients who complain of neck pain. AB - INTRODUCTION: Although neck pain is common in patients with chronic rheumatic disorders, any change in the quality or severity of pain should be further investigated. MATERIALS AND METHODS: We report two patients who recently presented with increasing neck pain on a background of chronic inflammatory arthritis. RESULTS: Both patients were found to have suffered fractures of the cervical spine. In each case, the bones appeared osteopenic and reduced bone density is likely to have made the patients susceptible to fracture. CONCLUSION: This report highlights the need to comprehensively evaluate patients with chronic rheumatic disorders who note a significant change in their neck symptoms. PMID- 19129981 TI - Ectopic pituitary adenoma presenting as midline nasopharyngeal mass. AB - INTRODUCTION: Ectopic pituitary adenomas are extremely rare. We report a case of ectopic pituitary adenoma in the midline of the nasopharynx. This adenoma probably arose from the pharyngeal remnant of Rathke's pouch. METHODS: We discuss a case of a lady who presented to our unit with 2 months history of dryness and sensation of lump in her throat and a long standing history of hypothyroidism. Examination of nasopharynx revealed a smooth and fluctuant midline mass. CT scan of nose and paranasal sinuses confirmed the midline mass with small defect communicating with the sphenoid sinus. An initial diagnosis of Thornwaldt's cyst was made and she underwent upper aerodigestive tract endoscopy and marsupialization of the mass. Histopathological examination revealed ectopic pituitary adenoma. CONCLUSION: Ectopic pituitary adenoma is an important differential diagnosis for a midline nasopharyngeal mass. It is recommended that prior to surgical resection of midline nasopharyngeal mass biopsy is taken and MRI is performed. PMID- 19129982 TI - Analysis of 35 inorganic elements in teeth in relation to caries formation. AB - As caries is a most common affliction of teeth, which are predominantly composed of inorganic elements, it was considered worthwhile to determine their elemental composition and to find if it bears any relation to caries formation. For this purpose, 35 inorganic elements were determined in 15 non-carious and an equal number of carious whole teeth after their dissolution in nitric acid. The results showed that out of 35 elements, strontium was the only element found to be present in significantly lower amount in carious teeth, thus strongly suggesting that its deficiency may help form caries. The decrease in Ca, Mg, Ti, P, Li, Be, V, Ni, Zn, Nb, Ag, Cd, Sb, Ba, La , W, Pb, and Bi and slight increase in K, Cr, Cu, and As was not statistically significant. However, statistically significantly greater amount of boron, manganese, molybdenum, and fluorine in the carious teeth indicates towards their possible role in predisposing or causing dental caries. PMID- 19129983 TI - Discrimination of bullet types using analysis of lead isotopes deposited in gunshot entry wounds. AB - In order to discriminate bullet types used in firearms, of which the victims died, the authors investigated lead isotope ratios in gunshot entry wounds from nine lead (unjacketed) bullets, 15 semi-jacketed bullets, and 14 full-jacketed bullets by inductively coupled plasma-mass spectrometry. It was found that the lead isotope ratio of 207/206 in gunshot entry wounds was the highest with lead bullets, and it decreased in order from full-jacketed to semi-jacketed bullets. Lead isotope ratios of 208/206 or 208/207 to 207/206 at the gunshot entry wound were able to discriminate semi-jacketed bullets from lead and full-jacketed ones, but it was difficult to discriminate between lead and full-jacketed bullets. However, a combination of element and lead isotope ratio analyses in gunshot entry wounds enabled discrimination between lead, semi-jacketed, and full jacketed bullets. PMID- 19129984 TI - Phagocytosis, oxidative burst, and produced reactive species are affected by iron deficiency anemia and anemia of chronic diseases in elderly. AB - Iron and oxidative stress have a regulatory interplay. During the oxidative burst, phagocytic cells produce free radicals such as hypochlorous acid (HOCl). Nevertheless, scarce studies evaluated the effect of either iron deficiency anemia (IDA) or anemia of chronic disease (ACD) on phagocyte function in the elderly. The aim of the present study was to determine the oxidative burst, phagocytosis, and nitric oxide (*NO) and HOCl, reactive species produced by monocytes and neutrophils in elderly with ACD or IDA. Soluble transferrin receptor, serum ferritin, and soluble transferrin receptor/log ferritin (TfR-F) index determined the iron status. The study was constituted of 39 patients aged over 60 (28 women and 11 men) recruited from the Brazilian Public Health System. Oxidative burst fluorescence intensity per neutrophil in IDA group and HOCl generation in both ACD and IDA groups were found to be lower (p < 0.05). The percentages of neutrophils and monocytes expressing phagocytosis in ACD group were found to be higher (p < 0.05). There was an overproduction of *NO from monocytes, whereas the fundamental generation of HOCl appeared to be lower. Phagocytosis, oxidative burst, and *NO and HOCl production are involved in iron metabolism regulation in elderly patients with ACD and IDA. PMID- 19129985 TI - Potential ecological risk assessment of heavy metal pollution in sediments of the Yangtze River within the Wanzhou section, China. AB - A potential ecological risk assessment was conducted based on the analyzing results of the typical pollutants (Pb, Cu, Cr, Cd, and Zn) contents in the sediments of Yangtze River within the Wanzhou section using the index number techniques of single factor and Hakanson method for the quality status of the sediments. The results indicted that cadmium (Cd) had the largest pollution index and was the main pollution factor among the metals. The ecological risk sequence of the metals was Cd > Zn >Pb > Cu >Cr, while the sequence of the potential ecological risk posed by the metals was Cd > Pb > Cu> Zn >Cr. The index range of potential ecological risk was from 101.39 to 184.31, and the average index of potential ecological risk factors (RI) was 152.35. The Yangtze River within the Wanzhou section has a middle potential ecological risk. PMID- 19129986 TI - Revertant somatic mosaicism in the Wiskott-Aldrich syndrome. AB - Up to 11% of patients affected with Wiskott-Aldrich syndrome (WAS) have presented with somatic mosaicism due to spontaneous in vivo reversion to normal of the original mutation or second-site compensatory mutations that restored production of the WAS gene product. The reasons underlying the high prevalence of this phenomenon in WAS are unclear and may include strong selective advantage of revertant cells over mutated populations, abnormally high general mutation rate and/or increased susceptibility of specific WAS gene sequences to DNA polymerase errors. Additional studies in human samples and in vitro/in vivo models of the disease will likely yield further insights into the mechanisms responsible for the occurrence of revertant mosaicism in WAS and elucidate additional biological characteristics of the WAS gene and protein. PMID- 19129987 TI - Plasticobezoar-another new entity for Rapunzel syndrome. AB - Rapunzel syndrome- a form of trichobezoar occurs when a gastric trichobezoar extends beyond the pylorus into the bowel. We report two cases of Rapunzel syndrome, one of them caused by ingestion of plastic material, so as to name it as a 'Plasticobezoar'. To the best of our knowledge no Rapunzel syndrome due to this material has been reported so far. PMID- 19129989 TI - Antipyretic effect of ketoprofen. AB - OBJECTIVE: The aim of this study was to investigate the efficacy and side effect profile of ketoprofen as well as compliance with respect to the taste of the drug and compare these parameters with those of acetaminophen and ibuprofen. METHODS: A total of 301 patients between 1-14 years of age who applied to emergency rooms of three medical centers with the complaint of fever that required antipyretic therapy were included in the study. Fever was measured with the aid of a tympanic thermometer (Braun Kronberg 6014) and followed for 4-6 hours. The measurement was repeated at 30, 60, 120 minutes, and again 4-6 hours after the initial assessment. RESULTS: The mean age of the patients was 47.8+/-41.1 months. The patients randomly received 15 mg/kg/dose of acetaminophen (n=112 group 1), 0.5 mg/kg/dose of ketoprofen (n=105, group 2), or 10 mg/kg/dose of ibuprofen (n=84, group 3). Fever was 38.4+/-0.7 degrees C, 38.4+/-0.7 degrees C, and 38.5+/-0.5 degrees C at 30 minutes; 38.0+/-0.7 degrees C, 37.9+/-0.7 degrees C, and 38.0+/ 0.6 degrees C at 60 minutes (p>0.05), 37.7+/-0.6 degrees C, 37.6+/-0.7 degrees C, and 37.7+/-0.5 degrees C at 120 minutes (p>0.05); 37.5+/-0.7 degrees C, 37.3+/ 0.6 degrees C, and 37.4+/-0.6 degrees C at 4-6 hours after admission (p>0.05). The fever was significantly lower at 30, 60, and 120 minutes in all group s (p<0.05). Early vomiting after medication (<6 hours) was observed in 3.8%, 13.5%, and 9.6% whereas late vomiting (6-48 hours) occurred in 1.3%, 2.7%, and 5.8% respectively (p>0.05). Bad taste was expressed by 5.1%, 12.2%, and 5.8% early (<6 hours), and 3.9%, 8.1%, and 3.8% late (6-48 hours) (p>0.05). There were no differences between age groups for antipyretic effect, taste and adverse effect in three drugs (p>0.05). CONCLUSION: All three drugs were similar in terms of efficacy, adverse effects, and compliance within 48 hours of therapy. These results suggest that ketoprofen may be used for antipyresis as an alternative to acetaminophen and ibuprofen. PMID- 19129990 TI - Pulmonary actinomycosis. AB - Pulmonary actinomycosis is rarely reported in pediatric age. An 11-year-old girl with history of two-month back pain was admitted to our hospital. On physical examination respiratory sounds were diminished on the left upper lung. Chest radiograph revealed a mass in the left upper lobe. Computed tomography showed solitary lesion (5.6 x 4.5 cm in size) in the left upper lobe. We could not rule out the possibility of malignant thoracic tumor. The patient underwent surgery. Histological examination of the resected tissue revealed, numerous sulfur granules, characteristic of Actinomyces, surrounded by purulent exudates, which are consistent with actinomycosis. She was treated with penicillin G. The patient responded well to penicillin therapy and the lesions regressed completely. She remained well throughout the three-year follow-up. PMID- 19129992 TI - Management strategies in foreign-body aspiration. AB - OBJECTIVE: The aim of this study is to outline a management algorithm to ensure effective teamwork in decreasing morbidity and mortality in pediatric Foreign Body Aspirations (FBA). Furthermore, the role of flexible bronchoscopy when compared to rigid bronchoscopy in FBA was evaluated. METHODS: Charts of patients with suspected FBA from October 1999 to September 2006 were reviewed and data with regards to the history, presenting symptoms, diagnostics and therapeutic tactics, was collected. RESULTS: A total of 77 children with suspicion of FBA were managed in the 7 year period. Bronchoscopies were performed in 63 patients and in 26 foreign-bodies (FB) were found and extracted. At referral, 53 patients did not present acute respiratory symptoms, but had a positive history of FBA, and in 13 FB were found. Despite negative chest x-rays in 55 patients, FB were found in 8. Rigid bronchoscopy was performed in 53 and flexible in 10 patients. In 3 out of 10 patients who had undergone flexible bronchoscopy a FB was identified, the extraction of which was performed using a rigid bronchoscope. CONCLUSION: Clinical and radiological findings in children with typical history of suspected FBA are not enough to confirm the presence of FB. Successful management with an extremely low rate of morbidity and no mortality was observed using the algorithm used at our center. Flexible bronchoscopy reduces the chances of airway tract injury; however a rigid bronchoscope is necessary for FB removal. PMID- 19129991 TI - MRI findings in Joubert syndrome. AB - Joubert syndrome is a very rare autosomal recessive disorder with only 200 cases reported worldwide.Here we report 4 cases of this rare disorder with MRI findings. PMID- 19129993 TI - Massive bone marrow involvement by clear cell variant of rhabdomyosarcoma. AB - A 16-year-old boy with generalized myalgia and petechial hemorrhage was found to have a diffuse infiltrative disease in his bone marrow (BM). The BM aspirate contained a dense population of vacuolated blast-like cells. The BM biopsy displayed compact sheets of small round cells with clear cytoplasm, reminiscent of Ewing sarcoma. Immunostains were not diagnostically conclusive while transmission electron microscopy on the BM cells demonstrated a clear skeletal muscle differentiation. The morphologic findings led to a tentative designation of metastatic embryonal rhabdomyosarcoma (RMS). It was not until cytogenetic analysis revealed the specific translocation t(2;13)(q35;q14) did the alveolar RMS finally get confirmed. Despite an exhaustive search by imaging studies, a primary tumor was not detected. This case illustrates that the massive BM involvement by atypical alveolar RMS of unknown origin may pose serious diagnostic challenges. Multidisciplinary studies are required to reach a definitive diagnosis. PMID- 19129995 TI - Is there an association with constitutional structural chromosomal abnormalities and hematologic neoplastic process? A short review. AB - The occasional observation of constitutional chromosomal abnormalities in patients with a malignant disease has led to a number of studies on their potential role in cancer development. Investigations of families with hereditary cancers and constitutional chromosomal abnormalities have been key observations leading to the molecular identification of specific genes implicated in tumorigenesis. Large studies have been reported on the incidence of constitutional chromosomal aberrations in patients with hematologic malignancies, but they could not confirm an increased risk for hematologic malignancy among carriers of structural chromosomal changes. However, it is of particular interest that constitutional structural aberrations with breakpoints similar to leukemia associated specific breakpoints have been reported in patients with hematologic malignancies. Because of insufficient data, it remains still unclear if these aberrations represent random events or are associated with malignancy. There has been a substantial discussion about mechanisms involved in constitutional structural chromosomal changes in the literature. The documentation of more patients with constitutional structural chromosomal changes could be of major importance. Most importantly, the molecular investigation of chromosomal regions involved in rearrangements could give useful information on the genetic events underlying constitutional anomalies, contributing to isolation of genes important in the development of the neoplastic process. Regarding constitutional anomalies in patients with hematologic disorders, a survey of the cytogenetic data of our cytogenetics unit is herein also presented. PMID- 19129996 TI - Acute inferior myocardial infarction in a patient with severe haemophilia A disease. PMID- 19129997 TI - Association between p53 codon 72 genetic polymorphism and tobacco use and lung cancer risk. AB - Lung cancer (LCa) is the leading cause of death by cancer in men. Genetic and environmental factors play a synergistic role in its etiology. We explore in 111 lung cancer cases and 133 unrelated noncancer controls the gene-environment interaction (G x E) between p53cd72 polymorphism variants and smoking and the effect on LCa risk in two kinds of case-control designs. We assessed the interaction odds ratio (IOR) using an adjusted unconditional logistic model. We found a significant and positive interaction association between Pro* allele carriers and smoking habits in both case-control and case-only designs: IOR = 3.90 (95% confidence interval [CI] = 1.10-13.81) and 3.05 (95% CI = 1.63-5.72), respectively. These exploratory results suggest a synergistic effect of the smoking habit and the susceptibility of the Pro allele on lung cancer risk compared with each risk factor alone. PMID- 19129999 TI - Cognitive physiology of locomotion. PMID- 19130000 TI - Epidemiology of neurovascular damage. PMID- 19129998 TI - Olmesartan/Amlodipine: combination therapy for the treatment of hypertension [corrected]. AB - Hypertension is a highly prevalent disease and one of the most important modifiable risk factors for cardiovascular disease. Hypertension remains the leading cause of mortality and the third largest cause of disability in both developed and developing countries. Although recent guidelines and advisory statements are recommending lower thresholds and goals for antihypertensive treatment, approximately two thirds of patients do not achieve the goals. In the United States only 36.8% of hypertensive patients achieve the goal of <140/90 mmHg. Poor adherence to antihypertensive medication regimens contributes to the practice-outcome gap. In most hypertensive patients it is difficult or impossible to control blood pressure with one drug, thus current guidelines have recommended the use of combination therapy as first-line treatment, or early in the management of hypertension. Blocking two or more blood pressure regulatory systems provides a more effective and more physiologic reduction in blood pressure. Fixed-dose combinations offer many advantages over free-drug combinations, such as convenience of use, fewer adverse events, and greater antihypertensive potency. Similar to other combinations, fixed-dose combination tablets containing the dihydropyridine calcium channel blocker amlodipine and the angiotensin receptor blocker olmesartan bring together two distinct and complementary mechanisms of action, resulting in improved blood pressure control and potential for improved target organ protection relative to either class of agent alone. PMID- 19130001 TI - Neuroradiological studies in neurorehabilitation. PMID- 19130002 TI - SPECT and rehabilitation: which role? PMID- 19130003 TI - The predicting value of TCD in neurorehabilitation. PMID- 19130004 TI - Theoretically-driven approaches to motor learning. PMID- 19130005 TI - Arm function after stroke: outcomes, measures and recent therapeutical approaches. PMID- 19130006 TI - Rehabilitation and territory: organizational models. PMID- 19130008 TI - Neuroprotective strategies in acute ischemic stroke. PMID- 19130007 TI - Stroke and depression: clinical features and treatment. AB - CONCLUSION: Cerebrovascular diseases are associated with a high incidence of depressive disorder, but despite this high level of comorbidity, depression appears to go largely unrecognized and untreated. This problem may have serious consequences as depressive disorder worsens the prognosis eitherquoad vitam orquoad valetudinem, increases medical costs, and delays the return to work or to a normal social functioning. If previous treatments with traditional antidepressants such as TCAs were difficult in these patients because of the known cardiovascular and anticholinergic side effects, new antide-pressants (such as SSRIs, noradrenergic and specific serotonine antidepressants (NaSSAs), noradrenergic reuptake inhibitors (NARIs) may offer therapeutic advantages as they have little or no effect on cardiac conduction, only transient or no effect on orthostatic hypotension, and no effect on cognitive performances. PMID- 19130009 TI - Post-stroke seizures. PMID- 19130010 TI - Pragmatic disorders of verbal communication. AB - CONCLUSION: Verbal communication impairments frequently follow damage to the right cerebral hemisphere. These deficits are usually underestimated because RHD patients are not routinely referred to speech therapists. The nature of pragmatic disorders in verbal communication in RHD patients still remains to be clarified. In particular, it is still unclear whether these impairments are language specific or attention-specific (e.g. an attentional deficit with respect to verbal cues only, which hinders the ability to go beyond the literal meaning of sentences), or should to be interpreted as an expression of a wider impairment in processing complex materials. Further research is needed to properly quantify the incidence of verbal communication disorders in RHD patients and to understand how pragmatic competence is organized in the brain. PMID- 19130012 TI - Mechanisms of functional recovery following acute brain damage. PMID- 19130011 TI - Aphasia therapy. AB - CONCLUSION: From the point of view of rehabilitation, two groups of aphasic patients can be recognized. They differ in the severity of the aphasic disorder, in the type of evaluation that can and should be utilized to plan and monitor therapy, and in the therapeutic intervention itself. The therapeutic intervention obviously differs not only between the two groups of patients but also from patient to patient, since no two patients show exactly the same disorder. What I refer to here is a deeper difference, a difference that lies in the rationale underlying the choice of the different implementations. In the first group, the rationale originates in identification of the functional damage, based on an explicit model of the normal function and the choice of coherent and theoretically based exercises; in the second group, no cognitive analysis of the disorder is possible, the theoretical rationale (analysis of conversation) is coarse, and the therapeutic intervention is grossly similar from patient to patient. PMID- 19130013 TI - Disorders of space representation. PMID- 19130014 TI - The implementation of the helsingborg declaration on stroke management. PMID- 19130015 TI - The problem of rehabilitation and prevention in Italy. PMID- 19130016 TI - Ischemic stroke: acute therapy and principles of rehabilitation. PMID- 19130017 TI - Organization of stroke rehabilitation services. PMID- 19130018 TI - Pathophysiological approach to stroke therapy. PMID- 19130019 TI - Assessment of quality rehabilitation. PMID- 19130020 TI - Secondary prevention in patients with cerebrovascular ischaemic disease. PMID- 19130021 TI - Rehabilitation and secondary prevention in patients with cardiovascular diseases. PMID- 19130022 TI - Poster presentations. PMID- 19130023 TI - Guidelines on stroke management. PMID- 19130024 TI - Immune-mediated CNS damage. AB - Multiple sclerosis (MS) is a demyelinating autoimmune disease. However, the persisting neurological deficits in MS patients result from acute axonal injury and chronic neurodegeneration, which are both triggered by the autoreactive immune response. Innate immunity, mainly mediated by activated microglial cells and invading macrophages, appears to contribute to chronic neurodegeneration. Activated microglia produce several reactive oxygen species and proinflammatory cytokines which affect neuronal function, integrity and survival. Adaptive immunity, particularly in cytotoxic CD8+ T cells, participates in acute demyelination and axonal injury by directly attacking oligodendrocytes and possibly neurons as well. Understanding the mechanisms of immune-mediated neuronal damage might help to design novel therapy strategies for MS. PMID- 19130025 TI - Prospects for antigen-specific tolerance based therapies for the treatment of multiple sclerosis. AB - A primary focus in autoimmunity is the breakdown of central and peripheral tolerance resulting in the survival and eventual activation of autoreactive T cells. As CD4(+) T cells are key contributors to the underlying pathogenic mechanisms responsible for onset and progression of most autoimmune diseases, they are a logical target for therapeutic strategies. One method for restoring self-tolerance is to exploit the endogenous regulatory mechanisms that govern CD4(+) T cell activation. In this review, we discuss tolerance strategies with the common goal of inducing antigen (Ag)-specific tolerance. Emphasis is given to the use of peptide-specific tolerance strategies, focusing on ethylene carbodiimide (ECDI)-peptide-coupled cells (Ag-SP) and nonmitogenic anti-CD3, which specifically target the T cell receptor (TCR) in the absence of costimulatory signals. These approaches induce a TCR signal of insufficient strength to cause CD4(+) T cell activation and instead lead to functional T cell anergy/deletion and activation of Ag-specific induced regulatory T cells (iTregs) while avoiding generalized long-term immunosuppression. PMID- 19130027 TI - [Skin and eyes]. AB - Numerous diseases affect both skin and eyes due to similar ontogenetic origin. The eye is the second most common site of melanoma after the skin. The eyelids are predisposed for development of toxic and allergic dermatitis as the skin in this region is four times thinner than the other facial skin. The differential diagnosis must include atopic and seborrhoeic eyelid dermatitis. Atopic and vernal keratoconjunctivitis are associated with atopic eczema. Various immunobullous disorders involve the conjunctiva with varying severity. Side effects of dermatologic treatments with glucocorticoids, antimalarials, psoralens, retinoids, or tetracyclines may involve the eye. PMID- 19130026 TI - Potential triggers of MS. AB - MS is an immune mediated disease of the central nervous system (CNS) characterized by demyelination, axonal damage and neurologic disability. The primary cause of this CNS disease remains elusive. Here we will address our current understanding of the role of viruses as potential environmental triggers for MS. Virus infections can act peripherally (outside the CNS) or within the CNS. The association of viral infections with demyelinating disease, in both animals and humans, will be discussed, as will the potential contributions of peripheral infection with Torque Teno virus, infection outside of and/or within the CNS with Epstein-Barr virus and infection within the CNS with Human Herpesvirus 6 to MS. An experimental animal model, Theiler's murine encephalomyelitis virus infection of susceptible strains of mice is an example of viral infections of the CNS as a prerequisite for demyelination. Finally, the proposition that multiple virus infections are required, which first prime the immune system and then trigger the disease, as a model where infections outside of the CNS lead to inflammatory changes within the CNS, for the development of a MS-like disease is explored. PMID- 19130028 TI - Olfactory host finding, intermediate memory and its potential ecological adaptation in Nasonia vitripennis. AB - Associative learning of host-associated chemical cues was studied in Nasonia vitripennis, a parasitoid of fly pupae in nests of hole-nesting birds. When females encountered a fly pupa and performed one sequence of host recognition behaviour including drilling the ovipositor into the host in the presence of the artificial odour furfurylheptanoate (FFH), they were afterwards arrested by FFH in olfactometer experiments. The response vanished after 4 days and could be blocked after 3 days by feeding wasps with ethacrynic acid prior and after the training. This indicates the formation of an intermediate form of memory by one host experience in N. vitripennis. Interestingly, the trained wasps avoided odours that were not present during the host encounter, although naive wasps did not react to these odours. This unique behaviour probably causes wasps to focus during host searching on those chemical cues they have experienced in the host environment. Studies in nests of hole-nesting birds revealed that about 30% of all nests contained only one fly pupa, and laboratory studies showed that N. vitripennis females are able to parasitise around 100 fly pupae in their life. It is discussed that under these conditions, the formation of a non-permanent intermediate memory for host-associated odours after one host encounter is adaptive to avoid costs involved with formation and maintenance of memory for misleading cues. The demonstration of associative olfactory learning in N. vitripennis, the first parasitoid species with sequenced genome, opens the gate to study molecular mechanisms of memory formation and its ecological adaptation in parasitoids. PMID- 19130029 TI - Structure of genetic diversity in the two major gene pools of common bean (Phaseolus vulgaris L., Fabaceae). AB - Domesticated materials with well-known wild relatives provide an experimental system to reveal how human selection during cultivation affects genetic composition and adaptation to novel environments. In this paper, our goal was to elucidate how two geographically distinct domestication events modified the structure and level of genetic diversity in common bean. Specifically, we analyzed the genome-wide genetic composition at 26, mostly unlinked microsatellite loci in 349 accessions of wild and domesticated common bean from the Andean and Mesoamerican gene pools. Using a model-based approach, implemented in the software STRUCTURE, we identified nine wild or domesticated populations in common bean, including four of Andean and four of Mesoamerican origins. The ninth population was the putative wild ancestor of the species, which was classified as a Mesoamerican population. A neighbor-joining analysis and a principal coordinate analysis confirmed genetic relationships among accessions and populations observed with the STRUCTURE analysis. Geographic and genetic distances in wild populations were congruent with the exception of a few putative hybrids identified in this study, suggesting a predominant effect of isolation by distance. Domesticated common bean populations possessed lower genetic diversity, higher F(ST), and generally higher linkage disequilibrium (LD) than wild populations in both gene pools; their geographic distributions were less correlated with genetic distance, probably reflecting seed-based gene flow after domestication. The LD was reduced when analyzed in separate Andean and Mesoamerican germplasm samples. The Andean domesticated race Nueva Granada had the highest F(ST) value and widest geographic distribution compared to other domesticated races, suggesting a very recent origin or a selection event, presumably associated with a determinate growth habit, which predominates in this race. PMID- 19130030 TI - Use of selection with recurrent backcrossing and QTL mapping to identify loci contributing to southern leaf blight resistance in a highly resistant maize line. AB - B73 is a historically important maize line with excellent yield potential but high susceptibility to the foliar disease southern leaf blight (SLB). NC292 and NC330 are B73 near-isogenic lines (NILs) that are highly resistant to SLB. They were derived by repeated backcrossing of an elite source of SLB resistance (NC250P) to B73, with selection for SLB resistance among and within backcross families. The goal of this paper was to characterize the loci responsible for the increased SLB resistance of NC292 and NC330 and to determine how many of the SLB disease resistance quantitative trait loci (dQTL) were selected for in the development of NC292 and NC330. Genomic regions that differentiated NC292 and NC330 from B73 and which may contribute to NC292 and NC330s enhanced SLB resistance were identified. Ten NC250P-derived introgressions were identified in both the NC292 and NC330 genomes of which eight were shared between genomes. dQTL were mapped in two F(2:3) populations derived from lines very closely related to the original parents of NC292 and NC330--(B73rhm1 x NC250A and NC250A x B73). Nine SLB dQTL were mapped in the combined populations using combined SLB disease data over all locations (SLB AllLocs). Of these, four dQTL precisely colocalized with NC250P introgressions in bins 2.05-2.06, 3.03, 6.01, and 9.02 and three were identified near NC250P introgressions in bins 1.09, 5.05-5.06, and 10.03. Therefore the breeding program used to develop NC292 and NC330 was highly effective in selecting for multiple SLB resistance alleles. PMID- 19130031 TI - A major QTL conferring crown rot resistance in barley and its association with plant height. AB - Crown rot (CR) is one of the most destructive diseases of barley and wheat. Fusarium species causing CR survive in crop residue and a growing acceptance of stubble retention practices has exacerbated disease severity and yield loss. Growing resistant cultivars has long been recognised as the most effective way to reduce CR damage but these are not available in barley. In a routine screening of germplasm, a barley landrace from China gave the best CR resistance among the genotypes tested. Using a doubled haploid population derived from this landrace crossed to Franklin, we demonstrate that the CR resistance of TX9425 was conditioned by a major QTL. The QTL, designated as Qcrs.cpi-3H, was mapped near the centromere on the long arm of chromosome 3H. Its effect is highly significant, accounting for up to 63.3% of the phenotypic variation with a LOD value of 14.8. The location of Qcrs.cpi-3H was coincident with a major QTL conferring plant height (PH) and the effect of PH on CR reaction was also highly significant. When the effect of PH was accounted for by covariance analysis, the Qcrs.cpi-3H QTL remained highly significant, accounting for over 40% of the phenotypic variation. The existence of such a major QTL implies that breeding barley cultivars with enhanced CR resistance should be feasible. PMID- 19130032 TI - cDNA cloning, expression levels and gene mapping of photosynthetic and non photosynthetic ferredoxin genes in sunflower (Helianthus annuus L.). AB - Fatty acid desaturation in plastids and chloroplasts depends on the electron donor activity of ferredoxins. Using degenerate oligonucleotides designed from known photosynthetic and heterotrophic plant ferredoxin sequences, two full length ferredoxin cDNAs were cloned from sunflower (Helianthus annuus L.) leaves and developing seeds, HaFd1 and HaFd2, homologous to photosynthetic and non photosynthetic ferredoxins, respectively. Based on these cDNAs, the respective genomic sequences were obtained and the presence of DNA polymorphisms was investigated. Complete sequencing of the HaFd1 and HaFd2 genes in different lines indicated the presence of two haplotypes for HaFd2 and their alignment showed that sequence polymorphisms are restricted to the 5'-NTR intron. In addition, specific DNA markers for the HaFd1 and HaFd2 genes were developed that enabled the genes to be mapped. Accordingly, the HaFd1 locus maps to linkage group 10 of the public sunflower map, while the HaFd2 locus maps to linkage group 11. Both ferredoxins display different spatial-temporal patterns of expression. While HaFd2 is expressed at similar levels in all tissues tested (leaves, stem, roots, cotyledons and developing seeds), HaFd1 is more strongly expressed in green tissues than in all the other tissues tested. Both photosynthetic- and heterotrophic-ferredoxins are present in sunflower seeds and may contribute to fatty acid desaturation during oil accumulation. Nevertheless, the levels of HaFd2 expression during seed formation are distinct in lines that only varied in the HaFd2 haplotypes they expressed. PMID- 19130033 TI - Genetic loci associated with stem elongation and winter dormancy release in wheat. AB - In winter wheat (Triticum aestivum L.), the stem begins to elongate after the vernalization requirement is satisfied during winter and when favorable temperature and photoperiod conditions are attained in spring. In this study, we precisely measured elongation of the first extended internode on 96 recombinant inbred lines of a population that was generated from a cross between two winter wheat cultivars, Jagger (early stem elongation) and 2174 (late stem elongation). We mapped a major locus for stem elongation to the region where VRN-A1 resides in chromosome 5A. Visible assessment of winter dormancy release was concomitantly associated with this locus. VRN1 was previously cloned based on variation in vernalization requirement between spring wheat carrying a dominant Vrn-1 allele and winter wheat carrying a recessive vrn-1 allele. Both of two winter wheat cultivars in this study carry a recessive vrn-A1 allele; therefore, our results suggest that either VRN-A1 might invoke a new regulatory mechanism or a new gene residing close to VRN-A1 plays a regulatory role in winter wheat development. Phenotypic expression of the vrn-A1a allele of Jagger was more sensitive to the year of measurement of stem elongation than that of the vrn-A1b allele of 2174. In addition to QSte.osu.5A, several loci were also found to have minor effects on initial stem elongation of winter wheat. Seventeen of nineteen locally adapted cultivars in the southern Great Plaints contained the vrn-A1b allele. Hence, breeders in this area have inadvertently selected this allele, contributing to later stem elongation and more conducive developmental patterns for grain production. PMID- 19130034 TI - Mapping QTL for drought stress-induced premature senescence and maturity in cowpea [Vigna unguiculata (L.) Walp.]. AB - Cowpea is an important crop for subsistence farmers in arid regions of Africa, Asia, and South America. Efforts to develop cultivars with improved productivity under drought conditions are constrained by lack of molecular markers associated with drought tolerance. Here, we report the mapping of 12 quantitative trait loci (QTL) associated with seedling drought tolerance and maturity in a cowpea recombinant inbred (RIL) population. One hundred and twenty-seven F(8) RILs developed from a cross between IT93K503-1 and CB46 were screened with 62 EcoR1 and Mse1 primer combinations to generate 306 amplified fragment length polymorphisms for use in genetic linkage mapping. The same population was phenotyped for maintenance of stem greenness (stg) and recovery dry weight (rdw) after drought stress in six greenhouse experiments. In field experiments conducted over 3 years, visual ratings and dry weights were used to phenotype drought stress-induced premature senescence in the RIL population. Kruskall Wallis and multiple-QTL model mapping analysis were used to identify QTL associated with drought response phenotypes. Observed QTL were highly reproducible between stg and rdw under greenhouse conditions. Field studies confirmed all ten drought-response QTL observed under greenhouse conditions. Regions harboring drought-related QTL were observed on linkage groups 1, 2, 3, 5, 6, 7, 9, and 10 accounting for between 4.7 and 24.2% of the phenotypic variance (R(2)). Further, two QTL for maturity (R(2) = 14.4-28.9% and R(2) = 11.7-25.2%) mapped on linkage groups 7 and 8 separately from drought-related QTL. These results provide a platform for identification of genetic determinants of seedling drought tolerance in cowpea. PMID- 19130035 TI - Not for the eyes only: PAX6 and glucose metabolism. PMID- 19130036 TI - Antidepressant use before and after initiation of diabetes mellitus treatment. AB - AIMS/HYPOTHESIS: Although current literature suggests an association between diabetes and depression, the direction of the association is unclear. We examined the temporal association between diabetes and depression by studying antidepressant and benzodiazepine use around the initiation of diabetes treatment. METHODS: From a pharmacy registry database we selected 49,593 diabetic patients and a random sample of non-diabetic individuals (n = 154,441), all >40 years old. Antidepressant and benzodiazepine use was calculated for the 7 years before and 7 years after the index date. The index date in diabetes patients was defined as the date of initiation of diabetes medication. A random index date was assigned to non-diabetic individuals. Time-specific incidence rate ratios of antidepressant and benzodiazepine use were calculated for intervals of 1 year, 3 months and 1 month. RESULTS: Antidepressant and benzodiazepine use was increased 2 months before and 3 months after the initiation of diabetes treatment compared with non-diabetic individuals. The strongest increase in incidence of antidepressant and benzodiazepine use was seen in the month after initiation of diabetes treatment with incidence rate ratios of 2.4 (95% CI 2.0-3.0) and 3.4 (95% CI 3.0-3.8) respectively, after adjustment for age, sex and Chronic Disease Score. CONCLUSIONS/INTERPRETATION: The increased incidence of antidepressant and benzodiazepine use may be a consequence of the burden of disease, of starting with diabetes medication or of being diagnosed with diabetes. Our findings could also reflect earlier detection by their physician. PMID- 19130038 TI - Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system. AB - AIMS/HYPOTHESIS: Loss of the trophic support provided by surrounding non endocrine pancreatic cell populations underlies the decline in beta cell mass and insulin secretory function observed in human islets following isolation and culture. This study sought to determine whether restoration of regulatory influences mediated by ductal epithelial cells promotes sustained beta cell function in vitro. METHODS: Human islets were isolated according to existing protocols. Ductal epithelial cells were harvested from the exocrine tissue remaining after islet isolation, expanded in monolayer culture and characterised using fluorescence immunocytochemistry. The two cell types were co-cultured under conventional static culture conditions or within a rotational cell culture system. The effect of co-culture on islet structural integrity, beta cell mass and insulin secretory capacity was observed for 10 days following isolation. RESULTS: Human islets maintained under conventional culture conditions exhibited a characteristic loss in structural integrity and functional viability as indicated by a diminution of glucose responsiveness. By contrast, co-culture of islets with ductal epithelial cells led to preserved islet morphology and sustained beta cell function, most evident in co-cultures held within the rotational cell culture system, which showed a significantly (p < 0.05) greater insulin secretory response to elevated glucose compared with control islets. Similarly, insulin/protein ratio data suggested that the presence of ductal epithelial cells is beneficial for the maintenance of beta cell mass. CONCLUSIONS/INTERPRETATION: The data indicate a supportive role for ductal epithelial cells in islet viability. Further characterisation of the regulatory influences may lead to novel strategies to improve long-term beta cell function both in vitro and following islet transplantation. PMID- 19130039 TI - Continuous relationships between non-diabetic hyperglycaemia and both cardiovascular disease and all-cause mortality: the Australian Diabetes, Obesity, and Lifestyle (AusDiab) study. AB - AIMS/HYPOTHESIS: Hyperglycaemia is a risk factor for cardiovascular disease (CVD) and all-cause mortality in individuals without diabetes. We investigated: (1) whether the risk of all-cause and CVD mortality extended continuously throughout the range of fasting plasma glucose (FPG), 2 h plasma glucose (2hPG) and HbA(1c) values; and (2) the ability of these measures to improve risk prediction for mortality. METHODS: Data on 10,026 people aged >or=25 years without diagnosed diabetes were obtained from the population-based Australian Diabetes, Obesity and Lifestyle study. Between 1999 and 2000, FPG, 2hPG and HbA(1c) were assessed and all-cause (332 deaths) and CVD (88 deaths) mortality were obtained after 7 years. RESULTS: Both 2hPG and HbA(1c) exhibited linear relationships with all-cause and CVD mortality, whereas FPG showed J-shaped relationships. The adjusted HR (95% CI) for all-cause mortality per SD increase was 1.2 (1.1-1.3) for 2hPG and 1.1 (1.0-1.2) for HbA(1c). The HR for FPG <5.1 mmol/l (per SD decrease) was 2.0 (1.3 3.0); for FPG >or=5.1 mmol/l (per SD increase) the HR was 1.1 (1.0-1.2). Corresponding HRs for CVD mortality were 1.2 (1.0-1.4), 1.2 (1.0-1.3), 4.0 (2.1 7.6) and 1.3 (1.1-1.4). The discriminative ability of each measure was similar; no measure substantially improved individual risk identification over traditional risk factors. CONCLUSIONS/INTERPRETATION: In individuals without diagnosed diabetes, 2hPG and FPG, but not HbA(1c) were significant predictors of all-cause mortality, whereas all measures were significant predictors of CVD mortality. However, these glucose measures did not substantially improve individual risk identification. PMID- 19130040 TI - Childhood BMI trajectories and the risk of developing young adult-onset diabetes. AB - AIMS/HYPOTHESIS: The aim of this study was to examine the effects of childhood BMI growth dynamics on the risk of developing young adult-onset type 1 and type 2 diabetes. METHODS: Finnish national healthcare registers were used to identify individuals with diabetes diagnosed between 1992 and 1996 at 15-39 years of age. Non-diabetic control participants were chosen from the National Population Registry. Anthropometric measurements were obtained from the original child welfare clinic records. Only the case-control pairs with sufficient growth data recorded were included in the analyses (218/1,388 for type 1 diabetes [16%] and 64/1,121 for type 2 diabetes [6%]). Two developmental stages in BMI growth (the points of infancy maximum BMI and the BMI rebound) were examined, and conditional logistic regression was applied to the variables of interest. RESULTS: The risk for type 1 diabetes increased 1.19-fold per 1 kg/m(2) rise in the infancy maximum BMI (p = 0.02). In addition, there was a 1.77-fold increase in the risk for type 2 diabetes per 1 kg/m(2) rise in the level of BMI at the BMI rebound (p = 0.04). Higher values of BMI at these points corresponded to a larger BMI gain from birth to that developmental stage. Age at the infancy maximum BMI or age at the BMI rebound did not affect the risk for either type of diabetes. CONCLUSIONS/INTERPRETATION: The BMI gain in infancy among individuals who subsequently developed young adult-onset type 1 diabetes was faster than that of those who remained healthy. The excess BMI gain in individuals who developed young adult-onset type 2 diabetes could already be seen during early childhood. PMID- 19130042 TI - [Dynamic instrumentation techniques in early-onset scoliosis]. AB - Similar to the situation in idiopathic scoliosis, the decision for operative or conservative treatment is based on the progression of the curve. Apart from age and Cobb angle, the rib-vertebral angle difference and rotation of the vertebrae are decisive for the prognosis. Surgery is indicated if, despite consistent brace treatment, a progression of >10 degrees or a Cobb angle of more than 45 degrees is found. Two different dorsal dynamic systems are presently used. With the growing rod technique, one or two rods are fixed to the spine cranially and caudally and are repeatedly"lengthened." Surgery is mandatory at least every 6 months. The Orthobiom system follows a new approach in which a rigid fixation is done in the middle of the curve, while the ends of the rods are linked to flexible connectors that can slide while the child grows. No long-time results are presently available for either system, so no general recommendations can be given. PMID- 19130043 TI - [Complication management after unsuccessful operative shoulder stabilization]. AB - Operative repair of shoulder instability may fail because of multiple causes: a constitutional predisposition, a new trauma, incorrect diagnosis, inadequate operative techniques or inappropriate rehabilitation can be involved. The key to successful revision surgery is a thorough analysis of errors of the primary repair and the revision also has to deal with the decisive pathological factors. The management of revision surgery after failed surgery for patients with instability has to focus on the decisive pathological factors and has to take a higher complication rate and lower success rate than primary repairs into account.In agreement with the literature a retrospective investigation of 61 open revision surgeries after an average follow up of more 4 years showed recurrent dislocations in 6 patients (9.8%). One of these patients had an adequate trauma and a seizure as the cause of dislocation. After thorough examination three patients revealed signs of a primarily overlooked connective tissue disorder. PMID- 19130044 TI - Point-of-care testing of proteins. AB - Point-of-care testing (POCT) is a fast developing area in clinical diagnostics that is considered to be one of the main driving forces for the future in vitro diagnostic market. POCT means decentralized testing at the site of patient care. The most important POCT devices are handheld blood glucose sensors. In some of these sensors, after the application of less than 1 microl whole blood, the results are displayed in less than 10 s. For protein determination, the most commonly used devices are based on lateral flow technology. Although these devices are convenient to use, the results are often only qualitative or semiquantitative. The review will illuminate some of the current methods employed in POCT for proteins and will discuss the outlook for techniques (e.g., electrochemical immunosensors) that could have a great impact on future POCT of proteins. PMID- 19130041 TI - Anti-inflammatory effect of lifestyle changes in the Finnish Diabetes Prevention Study. AB - AIMS/HYPOTHESIS: Subclinical inflammation confers an increased risk of type 2 diabetes, cardiovascular disease, neurodegenerative disorders and other age related chronic diseases. Physical activity and diet can attenuate systemic immune activation, but it is not known which individual components of a comprehensive lifestyle intervention are most effective in targeting subclinical inflammation. METHODS: We used data from the baseline examination and the 1 year follow-up of a subsample of 406 of 522 participants of the Finnish Diabetes Prevention Study (DPS) to estimate the effect of individual components of lifestyle intervention on C-reactive protein (CRP) and IL-6 levels, which represent the best characterised proinflammatory risk factors for type 2 diabetes. Changes in metabolic markers, dietary patterns and exercise were analysed to determine which were most strongly associated with the anti inflammatory effect of lifestyle changes. RESULTS: Lifestyle intervention reduced circulating levels of CRP (p < 0.001) and IL-6 (p = 0.060). Increases in fibre intake and moderate to vigorous leisure time physical activity (LTPA), but not total LTPA, predicted decreases in CRP and/or IL-6 and remained associated even after adjustment for baseline BMI or changes in BMI during the first year of the study. Changes in carbohydrate or fat intake were either weakly or not linked to reductions in CRP and IL-6. CONCLUSIONS/INTERPRETATION: The present study assessed the individual effects of dietary and physical activity measures on low grade inflammation in individuals at high cardiometabolic risk. Our results underline the importance of moderate to vigorous LTPA and a diet rich in natural fibre, and this should be emphasised in lifestyle recommendations. PMID- 19130045 TI - Operation and application of a new time-of-flight e-gas secondary neutral mass spectrometer (ToF-SNMS). AB - The low-pressure rf plasma of a secondary neutral mass spectrometer (e-gas SNMS) was connected with a time-of-flight (ToF) mass spectrometer for the first time. As opposed to ToF-SIMS in e-gas SNMS, the primary ion pulse cannot be used for triggering the flight time measurement. Therefore, an extraction pulse is used which at a defined time loads an ion package from the beam of the post-ionised particles into the ToF spectrometer. The newly developed ToF-SNMS system is described, and first experimental results are presented. PMID- 19130047 TI - Persistent left superior vena cava. PMID- 19130046 TI - Ureteric guidewire damage by Holmium:YAG laser: preliminary results. AB - Typically, guidewires are regularly used to provide access or act as a guide during laser lithotripsy. This may result in the tip of the fibre being in close proximity to the guidewire during the firing period and consequently, this could result in accidental damage to the guidewires during the procedure. To replicate this scenario, an experimental model was designed enabling accurate and reproducible positioning of the laser fibre relative to the guidewire. The guidewire was exposed to the laser energy for a range of typical settings used in the clinic. The results demonstrate that the guidewire is susceptible to laser energy damage, especially in close proximity to the fibre. PMID- 19130048 TI - Location of the ovaries in girls from newborn to 18 years of age: reconsidering ovarian shielding. AB - BACKGROUND: Guidelines for ovarian shielding are to place a lead shield in the midline of the abdomen at the level of the umbilicus. However, the ovaries are routinely seen at other locations on all types of imaging examinations. OBJECTIVE: To determine the position of the ovaries in girls, newborn to 18 years of age, in order to assess efficacy of ovarian shield placement. MATERIALS AND METHODS: We identified 336 girls who underwent lumbar spine and pelvic MRI. Images were reviewed noting the position of the ovaries relative to anatomic landmarks: symphysis pubis, iliac crest and anterior superior iliac spine (ASIS). In 174 girls a total of 307 ovaries were visualized. The girls were divided into three age groups and analyzed together. Right and left ovaries were analyzed together. The mean, 95% confidence interval (CI), standard deviations and range were calculated. RESULTS: The ovaries lay at or below the iliac crest (the level of the umbilicus), most often just medial to the ASIS and above the pubic symphysis in girls of all ages. CONCLUSION: Current methods of shielding only the midline of the pelvis for the purpose of reducing radiation dose to the ovaries during radiographic imaging are ineffective given that the ovaries are almost always positioned laterally in the pelvis. Therefore current shielding techniques should be changed; lead ovarian shields should be placed in a lateral position or even abandoned if relevant anatomy will be obscured. PMID- 19130049 TI - Edible oil degradation by using yeast coculture of Rhodotorula pacifica ST3411 and Cryptococcus laurentii ST3412. AB - To develop a microbial treatment of edible oil-contaminated wastewater, microorganisms capable of rapidly degrading edible oil were screened. The screening study yielded a yeast coculture comprising Rhodotorula pacifica strain ST3411 and Cryptococcus laurentii strain ST3412. The coculture was able to degrade efficiently even at low contents of nitrogen ([NH(4)-N]=240 mg/L) and phosphorus sources ([PO(4)-P]=90 mg/L). The 24-h degradation rate of 3,000 ppm mixed oils (salad oil/lard/beef tallow, 1:1 w/w) at 20 degrees C was 39.8%+/-9.9% (means+/-standard deviations of eight replicates). The highest degradation rate was observed at 20 degrees C and pH 8. In a scaled-up experiment, the salad oil was rapidly degraded by the coculture from 671+/-52.0 to 143+/-96.7 ppm in 24 h, and the degradation rate was 79.4%+/-13.8% (means+/-standard deviations of three replicates). In addition, a repetitive degradation was observed with the cell growth by only pH adjustment without addition of the cells. PMID- 19130050 TI - Antibiotic resistance genes in water environment. AB - The use of antibiotics may accelerate the development of antibiotic resistance genes (ARGs) and bacteria which shade health risks to humans and animals. The emerging of ARGs in the water environment is becoming an increasing worldwide concern. Hundreds of various ARGs encoding resistance to a broad range of antibiotics have been found in microorganisms distributed not only in hospital wastewaters and animal production wastewaters, but also in sewage, wastewater treatment plants, surface water, groundwater, and even in drinking water. This review summarizes recently published information on the types, distributions, and horizontal transfer of ARGs in various aquatic environments, as well as the molecular methods used to detect environmental ARGs, including specific and multiplex PCR (polymerase chain reaction), real-time PCR, DNA sequencing, and hybridization based techniques. PMID- 19130051 TI - New protocol for the rapid quantification of exopolysaccharides in continuous culture systems of acidophilic bioleaching bacteria. AB - In this study, we investigate exopolysaccharide production by a bacterial consortium during the bioleaching of a cobaltiferrous pyrite. Whereas comparable studies have looked at exopolysaccharide production in batch systems, this study focuses on a continuous system comprising a series of four stirred bioreactors and reveals the difficulties in quantifying biomolecules in complex media such as bioleached samples. We also adapted the phenol/sulphuric acid method to take into account iron interference, thus establishing a new protocol for sugar quantification in bioleached samples characterised by low pH (1.4) and high iron concentration (2 g l(-1)). This allows sugar analysis without any prior sample preparation step; only a small amount of sample is needed (0.5 ml) and sample preparation is limited to a single filtration step. We found that free exopolysaccharides represented more than 80% of the total sugars in the bioreactors, probably because stirring creates abrasive conditions and detaches sugars bound to pyrite or bacteria and that they were produced mainly in the first two reactors where bioleaching activity was greatest. However, we could not establish any direct link between the measured exopolysaccharide concentration and bioleaching activity. Exopolysaccharides could have another role (protection against stress) in addition to that in bacterial attachment. PMID- 19130052 TI - Enhanced transformation of malachite green by laccase of Ganoderma lucidum in the presence of natural phenolic compounds. AB - In this study, we investigated the efficacy of phenolic extract of wheat bran and lignin-related phenolic compounds as natural redox mediators on laccase-mediated transformation of malachite green (MG) using purified laccase from the white-rot fungus Ganoderma lucidum. G. lucidum laccase was able to decolorize 40.7% MG dye (at 25 mg l(-1)) after 24 h of incubation. Whereas, the addition of phenolic extract of wheat bran enhanced the decolorization significantly (p<0.001) by two- to threefold than that of purified laccase alone. Among various natural phenolic compounds, acetovanillone, p-coumaric acid, ferulic acid, syringaldehyde, and vanillin were the most efficient mediators, as effective as the synthetic mediator 1-hydroxybenzotriazole. Characterization of MG transformation products by HPLC, UV-Vis, and liquid chromatography-mass spectrometry-electrospray ionization analysis revealed that N-demethylation was the key mechanism of decolorization of MG by laccase. Growth inhibition test based on mycelial growth inhibition of white rot fungus Phanerochaete chrysosporium revealed that treatment with laccase plus natural mediators effectively reduced the growth inhibitory levels of MG than that of untreated one. Among all the tested compounds, syringaldehyde showed the highest enhanced decolorization, as a consequence reduced growth inhibition was observed in syringaldehyde-treated samples. The results of the present study revealed that the natural phenolic compounds could alternatively be used as potential redox mediators for effective laccase-mediated decolorization of MG. PMID- 19130053 TI - Myeloablative radioimmunotherapy in conditioning prior to haematological stem cell transplantation: closing the gap between benefit and toxicity? AB - High-dose radio-/chemotherapy in the context of autologous and allogeneic haematopoietic stem cell transplantation is a double-edged sword. The requirement for dose intensification is linked to an increase in toxicity to noninvolved organs. Particularly for older patients and patients with comorbidities, efficient but toxicity-reduced schemes are needed. Myeloablative radioimmunotherapy is a targeted, internal radiotherapy that uses radiolabelled monoclonal antibodies (mAb) with affinity to the bone marrow. It involves the administration of high radiation doses (up to 30 Gy) to the bone marrow and spleen but without exposing radiosensitive organs to doses higher than 1-7 Gy. Added to conventional or intensity-reduced conditioning, myeloablative radioimmunotherapy may achieve a pronounced antileukaemic effect with tolerable toxicities. A rational and individual design of the ideal nuclide-antibody combination optimizes therapy. The anti-CD33, anti-CD45 and anti-CD66 mAbs appear to be ideal tracers so far. The beta-emitter (90)Y is coupled by DTPA and is the best nuclide for myeloablation. Approval trials for DTPA anti-CD66 mAb are underway in Europe, and in the near future these therapies may become applicable in practice. This review gives an overview of current myeloablative conditioning radioimmunotherapy. We discuss the selection of the optimal radioimmunoconjugate and discuss how radioimmunotherapy might be optimized in the future by individualization of therapy protocols. We also highlight the potential advantages of combination therapies. PMID- 19130054 TI - Multimodality molecular imaging--from target description to clinical studies. AB - This highlight lecture was presented at the closing session of the Annual Congress of the European Association of Nuclear Medicine (EANM) in Munich on 15 October 2008. The Congress was a great success: there were more than 4,000 participants, and 1,597 abstracts were submitted. Of these, 1,387 were accepted for oral or poster presentation, with a rejection rate of 14%. In this article a choice was made from 100 of the 500 lectures which received the highest scores by the scientific review panel. This article outlines the major findings and trends at the EANM 2008, and is only a brief summary of the large number of outstanding abstracts presented. Among the great number of oral and poster presentations covering nearly all fields of nuclear medicine some headlines have to be defined highlighting the development of nuclear medicine in the 21st century. This review focuses on the increasing impact of molecular and multimodality imaging in the field of nuclear medicine. In addition, the question may be asked as to whether the whole spectrum of nuclear medicine is nothing other than molecular imaging and therapy. Furthermore, molecular imaging will and has to go ahead to multimodality imaging. In view of this background the review was structured according to the single steps of molecular imaging, i.e. from target description to clinical studies. The following topics are addressed: targets, radiochemistry and radiopharmacy, devices and computer science, animals and preclinical evaluations, and patients and clinical evaluations. PMID- 19130055 TI - Selecting the optimal image segmentation strategy in the era of multitracer multimodality imaging: a critical step for image-guided radiation therapy. PMID- 19130056 TI - Identification of prognostic factors for chronicity in patients with low back pain: a review of screening instruments. AB - Low back pain (LBP) is currently the most prevalent and costly musculoskeletal problem in modern societies. Screening instruments for the identification of prognostic factors in LBP may help to identify patients with an unfavourable outcome. In this systematic review screening instruments published between 1970 and 2007 were identified by a literature search. Nine different instruments were analysed and their different items grouped into ten structures. Finally, the predictive effectiveness of these structures was examined for the dependent variables including "work status", "functional limitation", and "pain". The strongest predictors for "work status" were psychosocial and occupational structures, whereas for "functional limitation" and "pain" psychological structures were dominating. Psychological and occupational factors show a high reliability for the prognosis of patients with LBP. Screening instruments for the identification of prognostic factors in patients with LBP should include these factors as a minimum core set. PMID- 19130057 TI - Activation of ER stress and inhibition of EGFR N-glycosylation by tunicamycin enhances susceptibility of human non-small cell lung cancer cells to erlotinib. AB - PURPOSE: The epidermal growth factor receptor (EGFR), an N-glycosylated transmembrane protein, is the target of erlotinib, an orally bioavailable agent approved for treatment of patients with non-small cell lung cancer (NSCLC). In this study, we examined whether inhibition of EGFR N-glycosylation and stimulation of endoplasmic reticulum (ER) stress by tunicamycin enhances erlotinib-induced growth inhibition in NSCLC cell lines. METHODS: We examined the effects of tunicamycin and erlotinib on cytotoxicity of erlotinib-sensitive and resistant NSCLC cell lines, as well its effects on apoptotic pathways and on EGFR activation and subcellular localization. RESULTS: A minimally cytotoxic concentration of tunicamycin (1 microM) resulted in approximatey 2.6-2.9 fold and approximatey 6.8-13.5 fold increase in erlotinib-induced antiproliferative effects in sensitive (H322 and H358) and resistant cell lines (A549 and H1650), respectively. We found that tunicamycin generated an aglycosylated form of 130 kDa EGFR. Tunicamycin additionally affected EGFR activation and subcellular localization. Interestingly, the combination of tunicamycin and erlotinib caused more inhibitory effect on EGFR phosphorylation than that of erlotinib alone. Moreover, the combination induced apoptosis in H1650 cells through induction of CHOP expression, activation of caspase-12 and caspase-3, cleavage of PARP and bak, and down-regulation of anti-apoptotic proteins bcl-xL and survivin. CONCLUSIONS: Overall, our data demonstrate that tunicamycin significantly enhances the susceptibility of lung cancer cells to erlotinib, particularly sensitizing resistant cell lines to erlotinib, and that such sensitization may be associated with activation of the ER stress pathway and with inhibition of EGFR N glycosylation. PMID- 19130058 TI - [Pathoanatomical preparation and reporting of specimens from precancerous lesions and carcinomas of the vulva]. AB - On the basis of varying morphology and pathogenesis, two types of vulvar intraepithelial neoplasias (VIN) have been defined: the common type (approximately 98%), classic VIN, is characterised by strong association to high risk HPV infection (up to 90%), occurrence at younger age (median age 30-40 years) and multifocality. The differentiated (or simplex) type is rare (1%-2%) and is associated with older age (median age 65 years) and p53 alterations. It is usually diagnosed in combination with vulvar (keratinizing) squamous cell carcinoma. The classification currently preferred by the WHO in which VIN are classified into VIN 1-3 is to be replaced due to new data and according to a proposal by the International Society for the Study of Vulvovaginal Diseases (ISSVD) which eliminates VIN 1 and combines VIN 2 and 3 to VIN of common or, depending on histopathology, differentiated type. Prognostically relevant factors in vulvar cancer include stage of disease, inguinal lymph node involvement, size of metastatic deposits and presence of extracapsular extension, depth of invasion and distance of the tumor from resection margins. Tumor grade and the presence of lymphovascular space involvement are controversially discussed. PMID- 19130059 TI - Patterns of pulmonary function in smoking and nonsmoking patients with progressive systemic sclerosis. AB - Pulmonary involvement is a prominent feature in systemic sclerosis and a significant cause of morbidity and mortality. A restrictive ventilatory defect is typical and static lung volumes are usually reduced in patients with ILDs. The possibility of obstruction of small airways in progressive systemic sclerosis (SSc) has been suggested by widespread bronchiolectasis and peribronchial fibrosis noted at necropsy. A total of 46 patients with a diagnosis of SSc were retrospectively included in this study. Patients were classified according to their smoking status (never smokers, n = 34 and ex or current smokers, n = 12). Patients were also compared on the basis of the presence or not of an obstructive pattern on spirometry. The purpose of this study was to establish if SSc patients who are smokers have a different pattern of pulmonary function involvement. Our hypothesis was that smoking habit was not the only cause of air trapping and that the existence of small airway involvement secondary to SSc itself cannot be excluded. PMID- 19130060 TI - Morphological, physiological and behavioural evaluation of a 'Mice in Space' housing system. AB - Environmental conditions likely affect physiology and behaviour of mice used for life sciences research on Earth or in Space. Here, we analysed the effects of cage confinement on the weightbearing musculoskeletal system, behaviour and stress of wild-type mice (C57BL/6JRj, 30 g b.wt., total n = 24) housed for 25 days in a prototypical ground-based and fully automated life support habitat device called "Mice in Space" (MIS). Compared with control housing (individually ventilated cages) the MIS mice revealed no significant changes in soleus muscle size and myofiber distribution (type I vs. II) and quality of bone (3-D microarchitecture and mineralisation of calvaria, spine and femur) determined by confocal and micro-computed tomography. Corticosterone metabolism measured non invasively (faeces) monitored elevated adrenocortical activity at only start of the MIS cage confinement (day 1). Behavioural tests (i.e., grip strength, rotarod, L/D box, elevated plus-maze, open field, aggressiveness) performed subsequently revealed only minor changes in motor performance (MIS vs. controls). The MIS habitat will not, on its own, produce major effects that could confound interpretation of data induced by microgravity exposure during spaceflight. Our results may be even more helpful in developing multidisciplinary protocols with adequate scenarios addressing molecular to systems levels using mice of various genetic phenotypes in many laboratories. PMID- 19130061 TI - Quality of life: urinary bladder augmentation or substitution in children. AB - PURPOSE: Bladder augmentation and substitution has been assumed to improve health related quality of life in patients with urinary incontinence. This study was performed to elicit an evidence base for or against the above hypothesis. METHODS: Between 1988 and 2006, 67 bladder augmentations and 7 bladder substitutions were performed at our institute. Inclusion criteria for the cross sectional study were a postoperative period of more than 1 year and an age of at least 10 years at the time of operation. A multimodality treatment-specific questionnaire (comprising 38 questions) was designed and sent to 61 patients. Quality of life was investigated in all patients and between the groups of patients with meningomyelocele (Group A) versus bladder exstrophy (Group B), patients, who are catheterizing themselves via urethra (Group C) versus stoma (Group D) and patients who are using (Group E) versus not using wheelchair (Group F) following the surgery. For the statistical analysis Students t test, Wilcoxon signed rank test and correlation analysis were used. RESULTS: A significant overall improvement was found in patients quality of life following this surgery (P < 0.05). Ninety percent of patients would prefer again bladder augmentation or substitution to their previous state. Patients with meningomyelocele are changing pads or diapers more frequently than exstrophy patients because of their bowel problems postoperatively. Quality of life improved better in patients performing CIC via stoma than in patients who perform it via their native urethra (P < 0.05). Outcomes were independent of patients age and of the post-augmentation time to assessment (P < 0.05). CONCLUSIONS: Bladder augmentation or substitution significantly improved the health-related quality of life in children and young adolescents taking part in the study. The authors are planning a prospective long term follow-up of the patients (longitudinal study) to validate the results. PMID- 19130063 TI - PET-CT in detection of meningeal metastasis in neuroblastoma. AB - Neoplastic meningitis is being recognized with increasing frequency in patients with cancer: the common causes being adenocarcinomas originating from the lung, stomach, breast, ovary, malignant melanoma, leukemia, lymphoma, Ewings sarcoma, rhabdomyosarcoma, retinoblastoma and primary CNS malignancies. Meningeal metastases, though rare can be seen in advanced stages of neuroblastoma. Recognition of meningeal metastases is crucial for successful diagnosis and prompt treatment of these patients. Here, we present two patients of neuroblastoma in whom positron emission tomography-computed tomography (PET-CT) examination resulted in detection of meningeal metastases at diagnosis; thus, emphasizing the need of inclusion of brain imaging in PET-CT protocol in all cases of advanced neuroblastoma. PMID- 19130062 TI - Review of the evidence on the management of blunt renal trauma in pediatric patients. AB - Due to the size and location within the pediatric patient, the kidneys are susceptible to injury from blunt trauma. While it is clear that the goal of management of blunt renal trauma in children is renal preservation, the methods of achieving this goal have not been well established in the current literature. Therefore, we have set out to summarize and clarify the current published information on the management strategies for blunt renal trauma in children. While there is extensive literature available, it consists mostly of retrospective series documenting widely varied management styles. The purpose of this review is to display the current information available and delineate the role for future studies that may allow us to develop consistent management strategies of pediatric patients, who have sustained blunt renal trauma, in a safe and cost-effective manner. PMID- 19130064 TI - An oblique anastomosis has more linear length than a transverse anastomosis of a tubular structure in oesophageal anastomosis. AB - INTRODUCTION: Oesophageal stricture continues to be a challenging paediatric surgical problem. This study aimed to compare the results of oblique and transverse anastomosis to prevent anastomotic stricture that occurred following oesophageal repairs. MATERIALS AND METHODS: Twelve New Zealand rabbits were divided equally into two groups: oblique (O) and transverse (T). A 1-cm-long tract of the cervical oesophagus was resected through a cervical incision in both groups. Anastomosis was performed in both groups. Oesophagographic analysis was carried out on postoperative day 7 and the animals were fed orally on the same day on the condition that there was no oesophageal leakage. The rabbits were killed to measure diameters of the oesophageal lumen and bursting pressure (BP) in the anastomosis region 8 weeks later. RESULTS: The diameters of the oesophageal anastomosis lines (3.9 +/- 0.10 mm) in the O group were significantly greater than those (1.9 +/- 0.09 mm) in the T group (P < 0.05). The values of BP (189 +/- 10 mmHg) in the O group were also significantly higher than those (116 +/- 4 mmHg) in the T group (P < 0.05). CONCLUSIONS: Our results suggested that oblique anastomosis is a better surgical procedure for preventing oesophageal stricture, as shown by the increased diameters of oesophageal anastomosis lines and BP. PMID- 19130065 TI - Comparison of the Bishop score, body mass index and transvaginal cervical length in predicting the success of labor induction. AB - PURPOSE: To evaluate the role of ultrasonographic and various maternal and fetal parameters in predicting successful labor induction. METHODS: Body mass index, cervical length, dilatation, effacement, Bishop score, parity, maternal age and birth weight were evaluated in 189 singleton pregnant women at 37-42 weeks of gestation and having induction of labor. All underwent induction of labor with oxytocin. Body mass index was calculated using the formula weight (kg)/height(2) (m), cervical measurement was performed by transvaginal ultrasonography and Bishop score was determined by digital examination of cervix. RESULTS: Logistic regression analysis indicated that the cervical length and body mass index were independent variables in determining the risk of cesarean section (OR = 1.206, P = 0.000, CI 95% = 1.117-1.303; OR = 1.223, P = 0.007, CI 95% = 1.058-1.414 respectively). In multiple linear regression analysis, the effect of cervical length and body mass index on induction delivery interval was found to be statistically significant (t = 5.738, P = 0.000; t = 2.680, P = 0.009, respectively). ROC curve showed that the best parameter in predicting the risk of cesarean section was cervical length and that cervical length and body mass index were better parameters compared to the Bishop score (the areas under the curve are 0.819, 0.701 and 0.416, respectively). CONCLUSIONS: Body mass index and transvaginal cervical length were better predictors compared to the Bishop score in determining the success of labor induction. PMID- 19130066 TI - Surgical treatment of intractable postpartum hemorrhage and changing trends in modern obstetric perspective. AB - PURPOSE: To update the prevalence and risk factors of intractable postpartum hemorrhage (IPH) and evaluate the effectiveness of surgical treatment modalities. METHODS: Between January 2002 and January 2008, IPH diagnosis was made in 86 cases. They were evaluated retrospectively. RESULTS: Placental implantation abnormalities were the leading cause, responsible for 45.6% of cases. Organ preserving surgery (OPS) methods were utilized in 47 cases, with a success rate of 76.6%. Among these cases, 11 were proceeded to hysterectomy. Hysterectomy was performed in 45 cases as definitive treatment. CONCLUSIONS: Compression sutures were quite effective in controlling hemorrhage due to placenta accreta and previa. For women who are hemodynamically stable and desirous for future fertility, OPS modalities should be applied first. Shorter operating time, hospital stay and less blood transfusion were seen with subtotal type hysterectomy, so in case of nonbleeding lower uterine segment, subtotal type should be performed first. PMID- 19130067 TI - Primary ovarian adenomyoma in a woman with endometrial polyp: a case report and review of the literature. AB - BACKGROUND: An adenomyoma presenting outside the uterus is an extremely rare entity and only three cases of primary ovarian adenomyoma have been reported up to date. CASE REPORT: We report the fourth case of ovarian adenomyoma in a 45 year-old woman with an endometrial polyp. Transvaginal ultrasonography revealed a solitary endometrial polyp with an enlarged left ovary which appeared heterogenous with isoechoic and mildly hyperechoic pattern. CONCLUSION: Total abdominal hysterectomy and bilateral salpingo-oopherectomy was performed and histologic examination revealed an adenomyoma arising primarily in the ovary. PMID- 19130068 TI - Schwannoma of the tongue: two case reports and review of the literature. AB - The aim of this study was to describe clinicopathologic and radiographic features of two cases of schwannoma involving the oral tongue and to review the literature of this unusual clinical entity. Case reports with review of the pathologic, radiologic and clinical data for two patients with schwannoma of the tongue are reported. Review of the literature of case reports of schwannomas (neurilemmomas) of the tongue from 1955 to 2006 with analysis of the patient's age, gender, presenting symptom(s), tumor size, and surgical approach was undertaken. The two patients in our series presented with painless swelling of the tongue. Transoral excision was performed and pathologic examination confirmed the diagnosis of schwannoma in both the cases. A total of 126 cases of schwannoma of the tongue have been reported in the English literature over the past 51 years. Schwannomas of the tongue typically present in the third decade of life (33%), display no gender predilection (52.8% female; 47.2% male) and often present as a painless mass (69.6%). Schwannomas are likely to elicit distressing symptoms when they occur in the posterior one-third of the tongue (63.2 vs. 13.5%) or approach 3 cm in greatest dimension (33.0 vs. 18.2 mm). The vast majority of cases have been treated with transoral excision (94.8%). Recurrence after surgical excision has not been reported. Schwannoma of the tongue is a relatively rare tumor of the head and neck. Transoral resection allows for removal of this tumor in a manner that precludes recurrence, avoids causing morbidity of tongue function, and remains the standard approach for the treatment of the vast majority of these tumors. PMID- 19130069 TI - Functional effects of repeated pressure loads upon the tympanic membrane: mechanical stiffness measurements after simulated habitual sniffing. AB - In experimental studies it was found that otitis media causes stiffness loss in the tympanic membrane, possible precursors to retraction pockets and cholesteatoma. Besides otitis media habitual sniffing behaviour is associated with the development of retractions. The present study aims to test the hypothesis that repeated sniffing manoeuvre may cause not only structural, epithelial tympanic membrane changes presumed to be possible precursors to retractions, but also tympanic membrane stiffness loss, another possible mediator for the development of retractions. An experimental model with a pressure chamber was used to mimic the pressure conditions for the tympanic membrane in habitual sniffers' ears. The stiffness properties of twelve Mongolian gerbil tympanic membranes were measured with moire interferometry after varying time up to 12 days with repeated pressure loading. Three days later, lower overall displacement were obtained in two ears; after 7-12 days the displacement readings were normal. This study with maximum of 12 days of pressure loading did not verify the hypothesis that habitual "sniffing" impairs the stiffness of the tympanic membrane. PMID- 19130070 TI - Detection of Helicobacter pylori in adenotonsiller tissue specimens by rapid urease test and polymerase chain reaction. AB - In recent studies, there have been many arguments concerning Helicobacter pylori being reservoir in adenotonsillar tissue. In this study, our objective was to detect whether adenoid and/or tonsillar tissue of patients diagnosed with chronic adenotonsillitis was a reservoir for H. pylori. This study was performed with 47 patients with the diagnosis of chronic tonsillitits and adenoid hypertrophy. Helicobacter pylori was searched by rapid urease test (RUT) and polymerase chain reaction (PCR). Presence of H. pylori glmM gene (formerly named as ureC gene) was tested using ureC and ureC2 primers. Fifty-five specimens used in the study were made up of 35 adenoid and 20 tonsil tissues. Rapid urease test was positive in three (5.5%) specimens. Helicobacter pylori was not detected in any of the patients by PCR. Further studies are needed to clarify the possible role of H. pylori in upper aerodigestive tract diseases such adenotonsillitis. PMID- 19130071 TI - Cervical sympathetic chain schwannoma masquerading as a carotid body tumour with a postoperative complication of first-bite syndrome. AB - Carotid body tumours (CBT) are the most common tumours at the carotid bifurcation. Widening of the bifurcation is usually demonstrated on conventional angiography. This sign may also be produced by a schwannoma of the cervical sympathetic plexus. A 45-year-old patient presented with a neck mass. Investigations included contrast-enhanced CT, MRI and magnetic resonance arteriography with contrast enhancement. Radiologically, the mass was considered to be a CBT due to vascular enhancement and splaying of the internal and external carotid arteries. Intraoperatively, it was determined to be a cervical sympathetic chain schwannoma (CSCS). The patient had a postoperative complication of first-bite syndrome (FBS).Although rare, CSCS should be considered in the differential diagnosis for tumours at the carotid bifurcation. Damage to the sympathetic innervation to the parotid gland can result in severe postoperative pain characterised by FBS and should be considered in all patients undergoing surgery involving the parapharyngeal space. PMID- 19130072 TI - Endoscopic supraglottic laryngectomy: a proposal for a classification by the working committee on nomenclature, European Laryngological Society. AB - In July 1999, the European Laryngological Society (ELS) has accepted a proposal for the classification of different laryngeal endoscopic cordectomies. This is actually a common classification system used as a tool for surgical training, documentation and comparison of results. The same harmonization work is deemed necessary for the treatment of supraglottic lesions. The ELS is proposing a classification of the different laryngeal endoscopic supraglottic partial laryngectomies. This classification comprises four types of supraglottic laryngectomies: Type I, limited excision of small size superficial lesions of the free edge of the epiglottis, the ary-epiglottic fold, the arytenoid, or the ventricular fold or any other part of the supraglottis; Type II, medial supraglottic laryngectomy without resection of the pre-epiglottic space, suitable for T1 lesions of either the suprahyoid or the infrahyoid laryngeal surface of the epiglottis (Type IIa, superior hemi-epiglottectomy or Type IIb, total epiglottectomy, respectively); Type III, medial supraglottic laryngectomy with resection of the pre-epiglottic space, suitable for T1-T2 tumors of the infrahyoid endolaryngeal epiglottis without (Type IIIa) or with (Type IIIb) extension to the ventricular fold, necessitating its excision; finally, Type IV, lateral supraglottic laryngectomy, suitable for tumors of the threefolds' region, which may include the ventricular fold (Type IVa) or the arytenoid (Type IVb), when involved. As in the cases of endoscopic cordectomies, these operations are similarly classified according to the surgical approach used and the degree of resection completed in order to facilitate their use in daily clinical practice. PMID- 19130073 TI - Scleral punch method with topical mitomycin C for safe revision of failed deep sclerectomy in nanophthalmic uveal effusion syndrome. AB - PURPOSE: To describe a novel surgical technique for revision of deep sclerectomy surgery in nanophthalmic uveal effusion syndrome. METHODS: A 46-year-old woman with recessively inherited nanophthalmos and chronic uveal effusions underwent a revision of previous failed deep sclerectomy surgery. In the presence of scar tissue, revision of sclerectomies was greatly facilitated by the use of a Kelly scleral punch (Katena Products, Inc., Denville, NJ, USA). To inhibit further scarring, mitomycin-C (MMC) was applied topically in each quadrant posterior to the equator. RESULTS: The scleral punch allowed outward scleral dissection from the suprachoroidal space, and brisk enlargement of sclerectomies in all quadrants without perforation of choroidal tissue. Post-operative conjunctival wound healing appeared unimpeded by the posteriorly placed MMC sponges. Near-total resolution of the effusions had occurred by 3 months. CONCLUSIONS: The scleral punch provides a safe and effective method for constructing and revising full thickness sclerectomies in the nanophthalmic eye, minimizing the risk of choroidal trauma. The use of topical MMC may reduce the risk of late failure of trans-scleral outflow due to fibrosis. PMID- 19130074 TI - Pattern of deoxy[Hb+Mb] during ramp cycle exercise: influence of aerobic fitness status. AB - During ramp exercise the deoxy[Hb + Mb] pattern follows a sigmoid model [f(x) = f (0) + A/(1 + exp(-(-c +dx))], indicating a non-linear muscle blood flow (Qm)/oxygen uptake (VO25)-relationship. We hypothesised that in trained cyclists the sigmoid would display a rightward shift, due to an increased oxidative capacity and/or higher percentage of slow-twitch fibres. A total of 10 cyclists and 11 physically active students (PA students) performed a relative ramp exercise (+/-12 min) and a ramp25-exercise (25 W min(-1)). Deoxy[Hb + Mb] was measured at the M. Vastus Lateralis by NIRS, normalized to the total amplitude of the response and expressed as a function of absolute and relative (%peakP) work rate. The work rate corresponding to c/d (i.e.50% of the amplitude of the deoxy[Hb + Mb] response) was the only parameter of the sigmoid that differed significantly between cyclists (57.9 +/- 4.4% and 60.1 +/- 4.1%peakP in the relative and ramp25, respectively) and PAstudents (49.6 +/- 4.2% and 48.2 +/- 5.1%peakP, respectively), indicating a rightward shift of the sigmoid in the cyclists. These results suggest a change in the time course of C((a-v))O(2) as a function of aerobic fitness status. PMID- 19130075 TI - Functional assembly and purinergic activation of bestrophins. AB - Proteins of the bestrophin family produce Ca(2+)-activated Cl(-) currents and regulate voltage-gated Ca(2+) channels. Bestrophin 1 was first identified in the retinal pigment epithelium. Four human paralogs (hBest1-hBest4) exist, and for some bestrophins, dimeric and heterotetrameric structures have been proposed. Here, we demonstrate that hBest1-hBest4 induce Cl(-) conductances of different amplitudes when expressed in HEK293 cells and when activated through purinergic stimulation. hBest1 mutants that are known to cause autosomal dominant macular dystrophy (Best disease) did not produce a Cl(-) current. Bestrophins were colocalized and showed molecular and functional interaction in HEK293 cells, overexpressing hBest1 and hBest2 or hBest4. Interaction was confirmed in airway epithelial cells coexpressing endogenous bestrophins. A fraction of hBest2 and hBest4 was expressed in the membrane, while most of hBest1 was found in the endoplasmic reticulum. Nevertheless, hBest1 has a clear role for the adenosine triphosphate (ATP; or uridine triphosphate)-induced Cl(-) current in both HEK293 and Calu-3 cells. Since native epithelial tissues typically express several bestrophin paralogs, these proteins may exist as heterooligomeric structures. PMID- 19130076 TI - Satellite cell ablation attenuates short-term fast-to-slow fibre type transformations in rat fast-twitch skeletal muscle. AB - The purpose of this time-course study was to determine whether satellite cell ablation within rat tibialis anterior (TA) muscles exposed to short-term chronic low-frequency stimulation (CLFS) would limit fast-to-slow fibre type transformations. Satellite cells of the left TA were ablated by exposure to gamma irradiation before 1, 2, 5 or 10 days of CLFS and 1 week later where required. Control groups received only CLFS or a sham operation. Continuous infusion of 5 bromo-2'-deoxyuridine revealed that CLFS first induced an increase in satellite cell proliferation at 1 day, up to a maximum at 10 days over control (mean +/- SEM, 5.7 +/- 0.7 and 20.4 +/- 1.0 versus 1.5 +/- 0.2 mm(-2), respectively, P < 0.007) that was abolished by gamma-irradiation. Myosin heavy chain mRNA, immunohistochemical and sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses revealed CLFS-induced fast-to-slow fibre type transformation began at 5 days and continued at 10 days; in those muscles that were also exposed to gamma irradiation, attenuation occurred within the fast fibre population, and the final fast-twitch to slow-twitch adaptation did not occur. These findings indicate satellite cells play active and obligatory roles early on in the time course during skeletal muscle fibre type adaptations to CLFS. PMID- 19130077 TI - Regulation of the cellulose synthase-like gene family by light in the maize mesocotyl. AB - The cellulose synthase-like (ZmCSL) gene family of maize was annotated and its expression studied in the maize mesocotyl. A total of 28 full-length CSL genes and another 13 partial sequences were annotated; four are predicted to be pseudogenes. Maize has all of the CSL subfamilies that are present in rice, but the CSLC subfamily is expanded from 6 in rice to 12 in maize, and the CSLH subfamily might be reduced from 3 to 1. Unlike rice, maize has a gene in the CSLG subfamily, based on its sequence similarity to two genes annotated as CSLG in poplar. Light regulation of glycan synthase enzyme activities and CSL gene expression were analyzed in the mesocotyl. A Golgi-localized glucan synthase activity is reduced by ~50% 12 h after exposure to light. beta-1,4-Mannan synthase activity is reduced even more strongly (>85%), whereas beta-1,4-xylan synthase, callose synthase, and latent IDPase activity respond only slightly, if at all, to light. At least 17 of the CSL genes (42%) are expressed in the mesocotyl, of which four are up-regulated at least twofold, seven are down regulated at least twofold, and six are not affected by light. The results contribute to our understanding of the structure of the CSL gene family in an important food and biofuel plant, show that a large percentage of the CSL genes are expressed in the specialized tissues of the mesocotyl, and demonstrate that members of the CSL gene family are differentially subject to photobiological regulation. PMID- 19130078 TI - Overexpression of Thellungiella halophila H(+)-PPase (TsVP) in cotton enhances drought stress resistance of plants. AB - An H(+)-PPase gene, TsVP from Thellungiella halophila, was transferred into two cotton (Gossypium hirsutum) varieties (Lumianyan19 and Lumianyan 21) and southern and northern blotting analysis showed the foreign gene was integrated into the cotton genome and expressed. The measurement of isolated vacuolar membrane vesicles demonstrated that the transgenic plants had higher V-H(+)-PPase activity compared with wild-type plants (WT). Overexpressing TsVP in cotton improved shoot and root growth, and transgenic plants were much more resistant to osmotic/drought stress than the WT. Under drought stress conditions, transgenic plants had higher chlorophyll content, improved photosynthesis, higher relative water content of leaves and less cell membrane damage than WT. We ascribe these properties to improved root development and the lower solute potential resulting from higher solute content such as soluble sugars and free amino acids in the transgenic plants. In this study, the average seed cotton yields of transgenic plants from Lumianyan 19 and Lumianyan 21 were significantly increased compared with those of WT after exposing to drought stress for 21 days at flowering stage. The average seed cotton yields were 51 and 40% higher than in their WT counterparts, respectively. This study benefits efforts to improve cotton yields in arid and semiarid regions. PMID- 19130080 TI - Implementation intentions about nonfocal event-based prospective memory tasks. AB - Implementation intentions are detailed and systematic plans that are developed during intention formation. We compared two different implementation intentions to standard event-based prospective memory instructions using three different kinds of intentions. Two of these intentions involved nonfocal cues whereas the remaining intention was about specific, focal cues. Implementation intentions dramatically increased detection performance for the nonfocal intentions. Because the exact cues could not be specified during intention formation, we argue that cue salience and that strengthening the cue to target action association are not very viable mechanisms to explain all instances of the beneficial consequences of forming implementation intentions. PMID- 19130079 TI - Insight into the salt tolerance factors of a wild halophytic rice, Porteresia coarctata: a physiological and proteomic approach. AB - Salinity poses a serious threat to yield performance of cultivated rice in South Asian countries. To understand the mechanism of salt-tolerance of the wild halophytic rice, Porteresia coarctata in contrast to the salt-sensitive domesticated rice Oryza sativa, we have compared P. coarctata with the domesticated O. sativa rice varieties under salinity stress with respect to several physiological parameters and changes in leaf protein expression. P. coarctata showed a better growth performance and biomass under salinity stress. Relative water content was conserved in Porteresia during stress and sodium ion accumulation in leaves was comparatively lesser. Scanning electron microscopy revealed presence of two types of salt hairs on two leaf surfaces, each showing a different behaviour under stress. High salt stress for prolonged period also revealed accumulation of extruded NaCl crystals on leaf surface. Changes induced in leaf proteins were studied by two-dimensional gel electrophoresis and subsequent quantitative image analysis. Out of more than 700 protein spots reproducibly detected and analyzed, 60% spots showed significant changes under salinity. Many proteins showed steady patterns of up- or downregulation in response to salinity stress. Twenty protein spots were analyzed by MALDI-TOF, leading to identification of 16 proteins involved in osmolyte synthesis, photosystem functioning, RubisCO activation, cell wall synthesis and chaperone functions. We hypothesize that some of these proteins confer a physiological advantage on Porteresia under salinity, and suggest a pattern of salt tolerance strategies operative in salt-marsh grasses. In addition, such proteins may turn out to be potential targets for recombinant cloning and introgression in salt sensitive plants. PMID- 19130081 TI - Changes in BAER wave amplitudes in relation to total serum bilirubin level in term neonates. AB - INTRODUCTION: Whether the severity of bilirubin neurotoxicity is closely related to the level of total serum bilirubin (TSB) remains to be determined. MATERIALS AND METHODS: We studied the amplitudes of brainstem auditory evoked response (BAER) components in 83 term neonates with TSB >10 mg/dL to detect any differences in bilirubin ototoxic effect on the amplitudes between different levels of TSB. RESULTS AND DISCUSSION: Compared to age-matched normal controls, the amplitudes of BAER waves III and V were reduced significantly (P<0.01 and 0.001). The V/I and V/III amplitude ratios were also decreased significantly (P<0.001 and 0.01). Although all amplitudes tended to be lower at higher TSB levels than at lower levels, none of the amplitudes correlated significantly with the level of TSB. Neither the V/I amplitude ratio nor the V/III amplitude ratio correlated with the TSB. No significant differences were found in any BAER wave amplitudes among the TSB levels 11-15, 16-20 and >20 mg/dL. In the comparison of amplitude data between any two of the three TSB levels, only wave V amplitude showed significant difference between TSB levels 11-15 and >20 mg/dL (P < 0.05). CONCLUSION: BAER wave amplitudes were significantly reduced in neonates with hyperbilirubinemia. However, there was no close correlation between the degree of amplitude reduction and the level of TSB. These results indicate that bilirubin toxicity to the neonatal brain is not closely related to the level of TSB. PMID- 19130082 TI - Mucopolysaccharidosis VI: the Italian experience. AB - The current paper describes the natural history and management of mucopolysaccharidosis VI (MPS VI) in all patients currently diagnosed with the disease in Italy. Nine patients (5.5-14.4 years) were included in the data review in March 2008. Gestational and perinatal data were normal for all patients. Median age at diagnosis was 1.9 years. During the course of the disease, all patients developed coarsened facial features, short stature, heart valve disease, eye problems, musculoskeletal problems, hepatosplenomegaly and neurological abnormalities. All patients received rhASB enzyme replacement therapy (ERT) and showed improvement or stabilisation in clinical manifestations after onset of therapy. The most frequently reported improvements were increased joint mobility and reduced hepatosplenomegaly. No relevant safety issues of ERT were reported. In conclusion, patients in Italy with MPS VI are diagnosed early in life. All patients have access to ERT and appear to benefit from this therapy. PMID- 19130083 TI - Myositis ossificans circumscripta: a paediatric case and review of the literature. AB - Myositis ossificans circumscripta (MOC), characterised by non-neoplastic heterotopic bone formation in soft tissue and skeletal muscle, is rare in children. At onset, it is difficult to distinguish MOC from a musculoskeletal infection or neoplasm, particularly in absence of trauma, and a biopsy is frequently required. We describe an 11-year-old boy with MOC in his thigh. At admission, minor local trauma was referred, and clinical examination revealed a tender and painful mass in the left thigh. Hypothesising a periarticular infection, a wide-spectrum antibiotic and an anti-inflammatory drug were given. The magnetic resonance imaging revealed an enlarged mass with marked enhancement of the lesion and a central, irregular non-enhanced area due to necrosis. After incisional biopsy, the histopathological examination found immature osteoblasts inside striated muscle fibres, as well as proliferating fibroblasts, which are all compatible with MOC, thereby ruling out infection or malignancy. The CT scan showed calcified deposits arranged in a ring, thus confirming MOC; a radiological follow-up was suggested to detect the lesion's maturation. Three months after diagnosis, surgical excision was performed. The post-operative period was uneventful. Six months later, the child was in good condition, with no signs of recurrence. In the literature, 57 paediatric cases have been described; most of these had an acute course, required excision and then had a favourable evolution. Our case study confirms the good prognosis of MOC and underlines how this benign condition should be considered in children presenting a tender and painful soft tissue swelling. PMID- 19130084 TI - Validation of a Western Blot for the detection of anti-Trichinella spp. antibodies in domestic pigs. AB - Trichinellosis is a zoonotic disease in humans caused by Trichinella spp. According to international regulations and guidelines, serological surveillance can be used to demonstrate the absence of Trichinella spp. in a defined domestic pig population. Most enzyme-linked immunosorbent assay (ELISA) tests presently available do not yield 100% specificity, and therefore, a complementary test is needed to confirm the diagnosis of any initial ELISA seropositivity. The goal of the present study was to evaluate the sensitivity and specificity of a Western Blot assay based on somatic Trichinella spiralis muscle stage (L1) antigen using Bayesian modeling techniques. A total of 295 meat juice and serum samples from pigs negative for Trichinella larvae by artificial digestion, including 74 potentially cross-reactive sera of pigs with other nematode infections, and 93 meat juice samples from pigs infected with Trichinella larvae were included in the study. The diagnostic sensitivity and specificity of the Western Blot were ranged from 95.8% to 96.0% and from 99.5% to 99.6%, respectively. A sensitivity analysis showed that the model outcomes were hardly influenced by changes in the prior distributions, providing a high confidence in the outcomes of the models. This validation study demonstrated that the Western Blot is a suitable method to confirm samples that reacted positively in an initial ELISA. PMID- 19130085 TI - Finding advanced third-stage larvae of Gnathostoma turgidum Stossich, 1902 in Mexico from natural and experimental host and contributions to the life cycle description. AB - In order to clarify the role of Gnathostoma turgidum as an etiological agent involved in human gnathostomiasis in Mexico, establish the taxonomic identity of the advanced third-stage larvae (AdvL(3)), and contribute to the knowledge of its life cycle, experimental host infections, examination of potential natural hosts, and morphological comparisons were carried out. Examination of ten species of potential hosts at San Pedro las Playas and Tres Palos Lagoon in Guerrero state, Mexico revealed that two (Kinosternon integrum and Rana zweifeli) were infected by 15 AdvL(3) of G. turgidum. A specific identity was obtained comparing these larvae with those recovered from hosts experimentally infected. The AdvL(3) measured 1.6 mm in length, with two cervical papillae (both in 12th row) and an excretory pore on the 19th row. The average of cephalic hooklets, from first to fourth row, was 30.8, 34.0, 36.7, and 39.6, respectively. This is the first record of AdvL(3) of G. turgidum in America, and it represents a significant contribution for the understanding of the life cycle of this species. PMID- 19130086 TI - Further description of Cruzia tentaculata (Rudolphi, 1819) Travassos, 1917 (Nematoda: Cruzidae) by light and scanning electron microscopy. AB - Species of Cruzia are parasites of the large intestine of marsupials, reptiles, amphibians, and mammalians. Cruzia tentaculata specimens were collected from the large intestine of Didelphis marsupialis (Mammalia: Didelphidae) from Colombia (new geographical record) and from Brazil and analyzed by light and scanning electron microscopy. The morphology of males and females by light microscopy corroborated most of the previous description and the ultrastructure by scanning electron microscopy evidence: the topography of the cuticle, deirids, amphids, phasmids in both sexes, a pair of papillae near the vulva opening, and the number and location of male caudal papillae, adding new features for species identification only observed by this technique. PMID- 19130087 TI - Proteins of the Plasmodium falciparum two transmembrane Maurer's cleft protein family, PfMC-2TM, and the 130 kDa Maurer's cleft protein define different domains of the infected erythrocyte intramembranous network. AB - Plasmodium falciparum Maurer's clefts participate in the transport of macromolecules within the cytoplasm, including the transport of virulence proteins to the erythrocyte membrane surface. We identified a family of genes PfMC-2TM encoding transmembrane proteins located within the intramembranous network of the infected erythrocyte using monoclonal antibody SP1C1. The distribution of the PfMC-2TM protein family within domains of the network was investigated by colocalization and confocal microscopy studies using monoclonal antibody SP1C1 specific for PFMC-2TM and monoclonal antibody SP1A6 specific for the130 kDa Maurer's cleft protein. Peptide-specific antibodies were prepared against six peptides from different domains of PfMC-2TM and used with the Mabs, as well as known antibodies specific to Maurer's clefts proteins (ring-expressed protein and membrane-associated histidine-rich protein 1), the erythrocyte membrane protein 1 (PfEMP-1), and serine-rich antigen in colocalization studies. We show that PfMC-2TM is located in the Maurer's clefts throughout the intracellular blood stage, and immunoelectron microscopy shows domains of PfMC 2TM localized in the parasitophorous vacuole and parasitophorous vacuole membrane. The distribution of the 130 kDa Maurer's cleft protein changes from within the parasite to the clefts during intracellular development as the parasite matures from young trophozoite to segmented schizont. PMID- 19130088 TI - Identification of proteins that interact with catalytically active calcium dependent protein kinases from Arabidopsis. AB - Calcium-dependent protein kinases (CDPKs) are essential sensor-transducers of calcium signaling pathways in plants. Functional characterization of CDPKs is of great interest because they play important roles during growth, development, and in response to a wide range of environmental stimuli. The Arabidopsis genome encodes 34 CDPKs, but very few substrates of these enzymes have been identified. In this study, we exploited the unique characteristics of CDPKs to develop an efficient approach for the discovery of CDPK-interacting proteins. High throughput, semi-automated yeast two-hybrid interaction screens with two different cDNA libraries each containing 18 million prey clones were performed using catalytically impaired and constitutively active AtCPK4 and AtCPK11 variants as baits. The use of the constitutively active versions of the CPK baits improved the recovery of positive interacting proteins relative to the wild type kinase. Titration of interaction strength by growth under increasing concentrations of 3-aminotriazole (3-AT), a histidine analog and competitive inhibitor of the His3 gene product, confirmed these results. Possible mechanisms for this observed improvement are discussed. The reproducibility of this approach was assessed by the overlap of several interacting proteins of AtCPK4 and AtCPK11 and the recovery of several putative substrates and indicated that yeast two hybrid screens using constitutively active and/or catalytically impaired forms of CDPK provides a useful tool to identify potential substrates of the CDPK family and potentially the entire protein kinase superfamily. PMID- 19130089 TI - Genetic effects in the leukotriene biosynthesis pathway and association with atherosclerosis. AB - Leukotrienes are arachidonic acid derivatives long known for their inflammatory properties and their involvement with a number of human diseases, most particularly asthma. Recently, leukotriene-based inflammation has also been shown to play an important role in atherosclerosis: ALOX5AP and LTA4H, both genes in the leukotriene biosynthesis pathway, have individually been shown to be associated with various cardiovascular disease (CVD) phenotypes. To assess the role of the leukotriene pathway in CVD pathogenesis, we performed genetic association studies of ALOX5AP and LTA4H in a family based study of early onset coronary artery disease (EOCAD) (GENECARD, 1,101 families) and in a non-familial dataset of EOCAD (CATHGEN, 656 cases and 405 controls). We found weak to moderate association between single nucleotide polymorphisms (SNPs) in ALOX5AP and LTA4H with EOCAD. The previously reported four-SNP haplotype (HapA) in ALOX5AP showed association with EOCAD in CATHGEN (P = 0.02), while controlling for age, race and CVD risk factors. HapK, the previously reported ten-SNP haplotype in LTA4H was associated with EOCAD in CATHGEN (P = 0.04). Another previously reported four-SNP haplotype in ALOX5AP (HapB) was not significant in our sample (P = 0.39). The overall lack of (or weak) association of single SNPs as compared with the haplotype results demonstrates the need for analyzing multiple SNPs within each gene in such studies. Interestingly, we detected an association of SNPs in ALOX5 (P < 0.05), the target of ALOX5AP, with CVD. Using a pathway-based approach, we also detected statistical evidence for interactions among ALOX5, ALOX5AP and LTA4H using RNA expression data from a collection of freshly harvested human aortas with varying degrees of atherosclerosis. The GENECARD families did not demonstrate evidence for linkage or association with ALOX5, ALOX5AP or LTA4H. Our results support a modest role for the leukotriene pathway in atherosclerosis pathogenesis, reveal important genomic interactions within the pathway, and suggest the importance of using pathway-based modeling for evaluating the genomics of atherosclerosis susceptibility. PMID- 19130091 TI - Biotreatment of phenol-contaminated wastewater in a spiral packed-bed bioreactor. AB - A spiral packed-bed bioreactor inoculated with microorganisms obtained from activated sludge was used to conduct a feasibility study for phenol removal. The reactor was operated continuously at various phenol loadings ranging from 53 to 201.4 g m(-3) h(-1), and at different hydraulic retention times (HRT) in the range of 20-180 min to estimate the performance of the device. The results indicated that phenol removal efficiency ranging from 82.9 to 100% can be reached when the reactor is operated at an HRT of 1 h and a phenol loading of less than 111.9 g m(-3) h(-1). At an influent phenol concentration of 201.4 g m(-3), the removal efficiency increased from 18.6 to 76.9% with an increase in the HRT (20 120 min). For treatment of phenol in the reactor, the maximum biodegradation rate (V (m)) was 1.82 mg l(-1) min(-1); the half-saturation constant (K (s)), 34.95 mg l(-1). PMID- 19130090 TI - Analysis of the MTHFD1 promoter and risk of neural tube defects. AB - Genetic variants in MTHFD1 (5,10-methylenetetrahydrofolate dehydrogenase/5,10 methenyltetrahydrofolate cyclohydrolase/ 10-formyltetrahydrofolate synthetase), an important folate metabolic enzyme, are associated with a number of common diseases, including neural tube defects (NTDs). This study investigates the promoter of the human MTHFD1 gene in a bid to understand how this gene is controlled and regulated. Following a combination of in silico and molecular approaches, we report that MTHFD1 expression is controlled by a TATA-less, Initiator-less promoter and transcription is initiated at multiple start sites over a 126 bp region. We confirmed the presence of three database polymorphisms (dbSNP) by direct sequencing of the upstream region (rs1076991 C > T, rs8010584 G > A, rs4243628 G > T), with a fourth (dbSNP rs746488 A > T) not found to be polymorphic in our population and no novel polymorphisms identified. We demonstrate that a common SNP rs1076991 C > T within the window of transcriptional initiation exerts a significant effect on promoter activity in vitro. We investigated this SNP as a potential risk factor for NTDs in a large homogenous Irish population and determined that it is not an independent risk factor, but, it does increase both case (chi (2) = 11.06, P = 0.001) and maternal (chi (2) = 6.68, P = 0.01) risk when allele frequencies were analysed in combination with the previously identified disease-associated p.R653Q (c.1958 G > A; dbSNP rs2236225) polymorphism. These results provide the first insight into how MTHFD1 is regulated and further emphasise its importance during embryonic development. PMID- 19130092 TI - Enhanced fructooligosaccharides and inulinase production by a Xanthomonas campestris pv. phaseoli KM 24 mutant. AB - Xanthomonas campestris pv phaseoli produced an extracellular endoinulinase (9.24 +/- 0.03 U mL(-1)) in an optimized medium comprising of 3% sucrose and 2.5% tryptone. X. campestris pv. phaseoli was further subjected to ethylmethanesulfonate mutagenesis and the resulting mutant, X. campestris pv. phaseoli KM 24 demonstrated inulinase production of 22.09 +/- 0.03 U mL(-1) after 18 h, which was 2.4-fold higher than that of the wild type. Inulinase production by this mutant was scaled up using sucrose as a carbon source in a 5-L fermenter yielding maximum volumetric (21,865 U L(-1) h(-1)) and specific (119,025 U g(-1) h(-1)) productivities of inulinase after 18 h with an inulinase/invertase ratio of 2.6. A maximum FOS production of 11.9 g L(-1) h(-1) and specific productivity of 72 g g(-1) h(-1) FOS from inulin were observed in a fermenter, when the mutant was grown on medium containing 3% inulin and 2.5% tryptone. The detection of mono and oligosaccharides in inulin hydrolysates by TLC analysis indicated the presence of an endoinulinase. This mutant has potential for large-scale production of inulinase and fructooligosaccharides. PMID- 19130093 TI - Does dexmedetomidine reduce secondary damage after spinal cord injury? An experimental study. AB - The aim of this experimental study was to investigate the possible protective effect of dexmedetomidine (DEX) on traumatic spinal cord injury (SCI). Twenty-two New Zealand rabbits were divided into three groups: sham (no drug or operation, n = 6), Control [SCI + single dose of 1 mL saline intraperitoneally (i.p), after trauma; n = 8] and DEX (SCI + 1 microg/kg dexmedetomidine in 1 mL, i.p, after trauma, n = 8). Laminectomy was performed at T10 and balloon angioplasty catheter was applied extradurally. Four and 24 h after surgery, rabbits were evaluated by an independent observer according to the Tarlov scoring system. Blood, cerebrospinal fluid (CSF), tissue samples from spinal cord were taken for biochemical and histopathological evaluations. After 4 h of SCI, all animals in control or DEX treated groups became paraparesic. On the other hand, 24 h after SCI, partial improvements were observed in both control and DEX treated groups. Traumatic SCI leads to increase in the lipid peroxidation and decreases enzymatic or nonenzymatic endogenous antioxidative defense systems. Again, SCI leads to apoptosis in spinal cord. DEX treatment slightly prevented lipid peroxidation and augmented endogenous antioxidative defense systems in CSF or spinal cord tissue, but failed to prevent apoptosis or neurodeficit after traumatic SCI. Therefore, it could be suggested that treatment with dexmedetomidine does not produce beneficial results in SCI. PMID- 19130094 TI - Two-level contiguous cervical disc disease treated with peek cages packed with demineralized bone matrix: results of 3-year follow-up. AB - Interbody cages are widely used instruments for cervical fusion operations. Long term follow-up studies are needed to clarify if these devices are dependable. In this prospective study, 79 patients (42 women and 37 men) with a mean age of 51 years operated between January 2000 and December 2005 for treatment of degenerative cervical disc disease and spondylosis associated with radiculopathy or myelopathy were evaluated. Patients underwent two-level contiguous anterior cervical discectomy and fusion operations with standard anterior Smith-Robinson approach. To achieve fusion PEEK cages packed with demineralized bone matrix mixed with autologous blood were used. Clinical outcome was evaluated with Odom's criteria and results were evaluated as 'excellent', 'good', 'fair' and 'poor'. Spinal curves, mobility and fusion status were assessed with anterior-posterior and lateral (neutral, flexion and extension) radiographs obtained before surgery and at 3, 12, 24 and 36 months postoperatively. The Ishihara curvature index (ICI) was used for spinal curve evaluation. Lateral dynamic (flexion and extension) radiographs at postoperative 12th month revealed the fusion status classified as 1A, 1B, 2A and 2B. The radiological outcomes were classified as 'non-fusion' when 2B healing was observed, and as 'fusion' when 1A, 1B or 2A healing was observed at the levels subjected to surgery. According to Odom's criteria, clinical outcomes were classified as 'excellent' or 'good' in 69 patients (success rate: 87.3%). Eight patients were graded as 'fair' and two as 'poor'. Preoperative mean ICI was 10.4+/-3.72 and postoperative mean ICI was 10.1+/-3.14. The difference was statistically insignificant (P>0.05); therefore, preoperative lordosis was said to be preserved at final follow-up. Final fusion rate (Types 1A, 1B, and 2A) was 91.7% (145/158 levels). Radiological imaging showed no cage failure or dislodgement and reoperation due to non-fusion was not needed. PMID- 19130095 TI - Dynamical asymmetries in idiopathic scoliosis during forward and lateral initiation step. AB - Adolescent idiopathic scoliosis (AIS) is characterized by morphological trunk modifications acting on body mass distribution. Some specific biomechanical strategies during postural regulation have been reported. Given that spinal deformity is three-dimensional, some strategy analysis resulting from different stepping directions should lead to a better understanding of the dynamic adaptation of behaviour. The aim of this study is to identify dynamic strategies of AIS patients stepping in lateral and forward directions. Ten AIS patients with a right thoracic curve and 15 controlled volunteers have been tested. Ground reaction forces (GRF) have been recorded for right-limb stepping and for left limb stepping associated to forward and lateral directions. Force amplitudes, corresponding occurrences, impulses of stepping phases and an asymmetry index have been computed. Asymmetry and variability increased in the AIS group, compared to the control group, whatever the stepping direction is. Asymmetry for AIS patients systematically provides an increased left initiation GRF compared to a right initiation. Nevertheless, for both groups, lateral initiation shows the largest asymmetry index reported for a forward initiation. More precisely, adaptive dynamic strategies for the AIS group have been characterized by an asymmetry between right and left limbs for lateral and forward initiation. These results can be explained by the influence of scoliosis pathology on dynamic movements due to spinal deformity. A right thoracic curve leads to an extra weight on the limb, which needs to be moved; consequently, stepping initiation with the right limb was more challenging for patients than stepping with the left limb. For the AIS group, the observed variability can also depend on the ontogenesis of adaptive strategies. Lateral step initiation has to be considered as the most relevant paradigm to study scoliosis and may also serve as a clinical basis for treatment to analyse the dynamic postural control and asymmetry strategies of the scoliosis patient. PMID- 19130096 TI - The role of spinal concave-convex biases in the progression of idiopathic scoliosis. AB - Inadequate understanding of risk factors involved in the progression of idiopathic scoliosis restrains initial treatment to observation until the deformity shows signs of significant aggravation. The purpose of this analysis is to explore whether the concave-convex biases associated with scoliosis (local degeneration of the intervertebral discs, nucleus migration, and local increase in trabecular bone-mineral density of vertebral bodies) may be identified as progressive risk factors. Finite element models of a 26 degrees right thoracic scoliotic spine were constructed based on experimental and clinical observations that included growth dynamics governed by mechanical stimulus. Stress distribution over the vertebral growth plates, progression of Cobb angles, and vertebral wedging were explored in models with and without the biases of concave convex properties. The inclusion of the bias of concave-convex properties within the model both augmented the asymmetrical loading of the vertebral growth plates by up to 37% and further amplified the progression of Cobb angles and vertebral wedging by as much as 5.9 degrees and 0.8 degrees, respectively. Concave-convex biases are factors that influence the progression of scoliotic curves. Quantifying these parameters in a patient with scoliosis may further provide a better clinical assessment of the risk of progression. PMID- 19130097 TI - Lateral radiological evaluation of transarticular screw placement in the lower cervical spine. AB - This study assessed the ideal district of lateral radiograph in evaluation of transarticular screw placement in the lower cervical spine. To assess the ideal zone of lateral radiographs in determining the safe or hazardous locations of the screw tips during transarticular screw implantation in the lower cervical spine. Transarticular screw in the lower cervical spine had been used as an alternative technique to achieve posterior cervical spine stability. Injury to the spinal nerves caused by transarticular screws which are too long must be identified quickly to minimize the neurologic complication. No previous radiological study regarding evaluation of the transarticular screw placement using lateral radiographs has been reported. Twelve cervical spines were removed from embalmed cadavers. Four transarticular screw placements with Dalcanto's technique under direct visualization, including placement of the screw tip staying the ventral cortex, 2, 4 and 6 mm over-penetration of the ventral cortex, were performed on each specimen. Following each placement, a lateral radiograph was taken. Each vertebral body was divided vertically into four equal zones, and another equal zone posterior to the posterior border of the vertebral body was defined as Zone pre-1. The numbers of screw tips seen in each zone were quantified for each placement. Partitions of chi (2) method was used to evaluate the ideal zone on lateral radiograph for transarticular screw insertion. At C34 and C45, no significant difference was found between Zone pre-1 and Zone 1 (chi(2) = 0.18, P > 0.50), while there was significant difference between Zones 1 and 2 (chi(2) = 73.6, P < 0.005), as well as Zones 2 and 3 (chi(2) = 13.2, P < 0.005). At C56 and C67, No significant difference was found between Zones 2 and 3 (chi(2) = 0.25, P > 0.50), while there was significant difference between Zone pre-1 and Zone 1 (chi(2) = 66.2, P < 0.005), as well as Zones 1 and 2 (chi(2) = 10.5, P < 0.005). Ideal screw tip positions on lateral radiograph for transarticular screw by Dalcanto's technique should be in Zone 1 at C34 and C45, in Zone pre-1 at C56 and C67. If the screw tip was in Zones 3 and 4, the safe rate will be decreased significantly and it might be too deep and be dangerous. PMID- 19130098 TI - Scoliosis and spine involvement in fibrous dysplasia of bone. AB - Few studies focused on the prevalence of scoliosis and involvement of the spine in patients with fibrous dysplasia (FD) of bone. We examined for FD involvement of the spine and scoliosis in 56 patients affected by FD of bone. Fifty patients were part of a cohort reported in a multicentric study on FD promoted by European Pediatric Orthopedic Society (EPOS) in 1999, and six were new patients. There were 30 females and 26 males (mean age 12.5 years; range 1-42 years). Twenty three had monostotic FD, 9 polyostotic FD, and 24 McCune-Albright Syndrome (MAS). Scoliosis was observed in 11 cases of polyostotic FD and MAS (33.3%). In seven of the patients with scoliosis (63.3%) spine was involved by FD lesional tissue. FD lesions involved the thoracic or lumbar spine in all patients but one, where cervical spine was also affected. A correlation between scoliosis and either spinal (p<0.01) or pelvic lesions (p<0.05) and pelvic obliquity (p<0.01) was observed. Three of the 11 patients showed familiarity for scoliosis but in 2 of them spine was involved by FD. Scoliosis and spine involvement were never detected in monostotic FD. This study indicates that in FD patients with polyostotic disease (1) the prevalences of FD involvement of the spine and scoliosis are high enough to include spine in the clinico-radiographic survey of these patients, and (2) the involvement of the spine and pelvis by FD lesions and pelvic obliquity are important determinants in the occurrence of scoliosis. PMID- 19130099 TI - Electrostatic potential maps of damaged DNA studied by image analysis tools. 8 Oxoguanine and abasic site lesions. AB - Changes of electrostatic potential around the DNA molecule resulting from chemical modifications of nucleotides may play a role in enzymatic recognition of damaged sites. The electrostatic potential around the DNA fragments containing either the intact guanine-cytosine pair or 8-oxoguanine-cytosine or the guanine abasic site was projected on a cylindrical surface around the double helix. The 2D maps of EP of intact and damaged DNA fragments were compared using image analysis methods. Occurrence of abasic site and 8-oxoguanine lesions were found to be reflected in the EP maps. In the case of the 8-oxoguanine lesion, the two phosphate groups and countercations of the damaged strand are moved away from the lesion in opposite directions, whereas they are moved in the same direction in the case of the abasic site lesion. The characteristic features of 8-oxoguanine lesion might be identified in the major groove, whereas the features of abasic site lesion the minor groove. PMID- 19130100 TI - Adsorption into the MFI zeolite of aromatic molecule of biological relevance. Investigations by Monte Carlo simulations. AB - Adsorption of paracresol and water into the silicalite-1 (MFI) zeolite has been investigated using canonical and grand-canonical Monte Carlo simulations. The most stable sites of adsorption of paracresol are found to be located at the channel intersections. Grand-canonical simulations have shown that at low loading, water molecules adsorb preferably at the vicinity of paracresol molecules, whereas they are also located in the sinusoidal channels as the loading increases. In order to explain the experimental adsorption isotherm observed for the coadsorption of water and paracresol in the MFI zeolite we propose a new concept of apparent adsorption enthalpy that varies with the concentration of the solution. The mathematical expression for the apparent enthalpy is introduced in an adsorption isotherm model. We shall refer to this theoretical isotherm as a non-langmuirian isotherm. The non-linear expression for the apparent adsorption enthalpy accounts for a variable accessibility of the sites of adsorption with respect to the concentration of the solution. PMID- 19130101 TI - Cross-fostering diminishes song discrimination in zebra finches (Taeniopygia guttata). AB - Song-production, -discrimination, and -preferences in oscine birds are dually influenced by species identity and the ontogenetic environment. The cross fostering of a model species for recognition research, the zebra finch (Taeniopygia guttata) into heterospecific nests of the Bengalese finch (Lonchura striata vars. domestica) allows an exploration of the sensory limits of early development and the effects of species-specific acoustic cues upon song discrimination in adulthood. To quantify the song preferences of female and male normal-reared (control) and Bengalese finch fostered zebra finches, we recorded multiple behavioral measures, including spatial proximity, vocalization rates and response latency, during sequential song-playback choice-trials using both tutor species' songs and the songs of two other ecologically relevant Australian species, the owl finch (Taeniopygia bichenovii) and the star finch (Neochmia ruficauda). Response strength was variable between the different measures, but no differences were detected within the specific behavioral responses towards the song playbacks of the two sexes. Control subjects strongly preferred their own species' songs while Bengalese-fostered zebra finches exhibited reduced song discrimination between con-, tutor-, and heterospecific songs. Overall behavioral responsiveness was also modulated by social ontogeny. These results indicate a difference in the strength of preference for song that is dependent on the species identity of the rearing environment in oscine birds and illustrate the role of multiple behavioral measures and ecologically relevant stimulus species selection in behavioral research using zebra finches. PMID- 19130102 TI - The effect of development and individual differences in pointing comprehension of dogs. AB - In spite of the rather different procedures actually used in comparative studies to test the ability of different species to rely on the human pointing gesture, there is no debate on the high performance of dogs in such tasks. Very little is known, however, on the course through which they acquire this ability or the probable factors influencing the process. Important developmental questions have remained unsolved and also some methodological concerns should be addressed before we can convincingly argue for one interpretation or another. In this study we tested 180 dogs of different age (from 2 months to adults) to investigate their performance in the human distal momentary pointing gesture. The results, analyzed at both the group and the individual levels, showed no difference in the performance according to age, indicating that in dogs the comprehension of the human pointing may require only very limited and rapid early learning to fully develop. Interestingly, neither the keeping conditions nor the time spent in active interaction with the owner, and not even some special (agility) training for using human visual cues, had significant effect on the success and explained individual differences. The performance of the dogs was rather stable over time: during the 20 trials within a session and even when subsamples of different age were repeatedly tested. Considering that in spite of the general success at the group level, more than half of the dogs were not successful at the individual level, we revealed alternative "decision-making rules" other than following the pointing gesture of the experimenter. PMID- 19130103 TI - Frequency of paradoxical effect with caspofungin in Candida albicans. PMID- 19130104 TI - Detection of Ureaplasma biovars and polymerase chain reaction-based subtyping of Ureaplasma parvum in women with or without symptoms of genital infections. AB - Ureaplasma parvum colonises human mucosal surfaces, primarily in the urogenital and respiratory tracts, causing a wide spectrum of diseases, from non-gonococcal urethritis to pneumonitis in immunocompromised hosts. Although the basis for these diverse clinical outcomes is not yet understood, it has been suggested that only certain strains of these micro-organisms are disease-associated. The aim of this study was to determine the distribution of Ureaplasma biovars and U. parvum serovars and to estimate their possible association with age, absence of lactobacilli, clinical symptoms and antibiotic resistance. DNA was extracted by endocervical, vaginal and urethral samples obtained from 158 women positive for U. urealyticum by culture and were biotyped by polymerase chain reaction (PCR) targeting the multiple-banded gene. Parvo biovar (biovar 1) was found in 136 (86%) and T960 biovar (biovar 2) in 22 (14%) patients. Among the different serovars of U. parvum, we found that serovar 3/14 was present maximally in the 21 25-year-old age group, while T960 biovar was distributed with quite similar frequency in women of 26-30 and >40 years of age. In this study, U. parvum serovar 3/14 and T960 biovar were found to be significantly associated with symptomatic patients and a loss of lactobacilli, while, on the contrary, U. parvum serovar 6 was significantly correlated with asymptomatic women and normal vaginal flora. The most active antibiotic for the majority of Ureaplasma isolates was tetracycline. These preliminary data show the possibility of distinguishing between the more or less virulent strains of Ureaplasma, with important consequences for therapeutic treatment. PMID- 19130105 TI - Genetic diversity of methicillin-resistant Staphylococcus aureus in a tertiary hospital in The Netherlands between 2002 and 2006. AB - The aim of this study was to investigate the methicillin-resistant Staphylococcus aureus (MRSA) clones isolated in a Dutch university hospital, situated near the borders of Belgium and Germany, between 2002 and 2006. MRSA strains (n = 175) were characterized using spa and SCCmec typing. The presence of Panton Valentine leukocidin (PVL) was determined. Between 2002 and 2005, ST5-MRSA-IV was predominant, and the spa type of ST5-MRSA-IV changed from t002 to t447. ST5-MRSA I, ST5-MRSA-II, ST228-MRSA-I, and ST247-MRSA-I were also observed in this period. From 2004, the MRSA genetic background became more diverse, and in 2006, ST5-MRSA IV was only sporadically observed. From 2005, ST5-MRSA-II, ST8-MRSA-IV, ST22-MRSA IV, and ST45-MRSA-IV were increasingly observed. Several other MRSA clones, such as ST239-MRSA-III, were found sporadically. Four PVL-positive MRSA isolates were observed, associated with ST80-MRSA-IV and ST8-MRSA-IV. ST5-MRSA-I, ST5-MRSA-II, ST5-MRSA-IV, and ST228-MRSA-I have not been described previously in The Netherlands. PMID- 19130106 TI - Molecular epidemiology of Echovirus 6 in Greece. AB - The objective was to investigate the genetic relationships among Echovirus 6 (E6) strains circulating in Greece and to compare them with the respective strains from other geographic regions. Cerebrospinal fluid samples collected during the period 2006-2007 from 84 patients with aseptic meningitis or encephalitis were tested for a probable enteroviral infection. Two RT-PCRs amplifying overlapping regions of the VP1 gene were performed, while isolation procedures were applied in one third of cases. All PCR products were sequenced, and further phylogenetic analysis was performed for E6 strains. Enteroviruses were detected in 27 out of 84 cases (32.14%) and E6 was the predominant serotype (11 out of 27, 40.74%). Three distinct clades of Greek E6 sequences were seen in the phylogenetic tree: sequences of the present study were placed in clades A and B, while sequences of a former study in Greece were clustered in clade C. Sequences of clades A and C presented high genetic homology (>95%) with sequences from other countries, while sequences of clade B were unique, differing by more than 15% from all known E6 sequences. The most prevalent enterovirus in Greece during the period 2006-2007 was E6, and was associated with aseptic meningitis. A high degree of heterogeneity was observed among Greek E6 strains. PMID- 19130107 TI - The role of polysaccharide intercellular adhesin (PIA) in Staphylococcus epidermidis adhesion to host tissues and subsequent antibiotic tolerance. AB - The aim of this study was to determine the role of polysaccharide intercellular adhesin (PIA) in Staphylococcus epidermidis adhesion to host tissues and subsequent antibiotic tolerance. The adherence of S. epidermidis 1457 and the mutant defective in PIA production (1457-M10) to urinary epithelium and endothelium was estimated by colony counting. Minimum bactericidal concentration and mean reduction of cellular activity (XTT) following antibiotic exposure was determined for planktonic and adhered bacteria. S. epidermidis 1457 adhered to a greater extent to both cells than the mutant strain. The adhered strains had a significantly higher antimicrobial tolerance than their planktonic counterparts. The mutant strain was, in general, the most susceptible to the antibiotics assayed. In conclusion, PIA may influence S. epidermidis adherence to host tissues and their antimicrobial susceptibility. Initial adhesion may be the main step for the acquisition of resistance in S. epidermidis. PMID- 19130108 TI - The ERG in drug development: translation from animal models to human. PMID- 19130110 TI - Uptake and incorporation of pinolenic acid reduces n-6 polyunsaturated fatty acid and downstream prostaglandin formation in murine macrophage. AB - Many reports have shown the beneficial effects of consumption of pine seeds and pine seed oil. However, few studies have examined the biological effect of pinolenic acid (PNA; 5,9,12-18:3), the main fatty acid in pine seed oil. In this study, using murine macrophage RAW264.7 cells as a model, we examined the effect of PNA on polyunsaturated fatty acid (PUFA) metabolism, prostaglandin (PG) biosynthesis and cyclooxygenase-2 (COX-2) expression. Results showed that PNA was readily taken up, incorporated and elongated to form eicosatrienoic acid (ETrA, 7,11,14-20:3) in macrophage cells. A small portion of this elongated metabolite was further elongated to form 9,13,16-22:3. The degree of incorporation of PNA and its metabolites into cellular phospholipids varied with the length of incubation time and the concentration of PNA in the medium. Incubation of PNA also modified the fatty acid profile of phospholipids: the levels of 18- and 20 carbon PUFA were significantly decreased, whereas those of 22-carbon fatty acids increased. This finding suggests that PNA enhances the elongation of 20-carbon fatty acids to 22-carbon fatty acids. The syntheses of PGE(1) from dihomo-gamma linolenic acid (DGLA, 8,11,14-20:4) and PGE(2) from arachidonic acid (ARA, 5,8,11,14-20:4) were also suppressed by the presence of PNA and its metabolite. As the expression of COX-2 was not suppressed, the inhibitory effect of PNA on PG activity was attributed in part to substrate competition between the PNA metabolite (i.e., 7,11,14-20:3) and DGLA (or ARA). PMID- 19130109 TI - Influence of commercial dietary oils on lipid composition and testosterone production in interstitial cells isolated from rat testis. AB - The aim of this study was to examine the influence of dietary fat on lipid composition, as well as on the steroidogenic function of interstitial cells isolated from Wistar rats that had been fed semi-synthetic diets containing four different commercial oils (S soybean, O olive, C coconut, and G grape seed). Steroidogenic enzyme activities, lipid composition, and androgen production were measured in testicular interstitial cells. Lipid analysis included measurement of the contents of major lipid subclasses (neutral lipids, polar lipids, free and esterified cholesterol), as well as principal polar and neutral lipid fatty acyl compositions. Significant differences in lipid composition were observed among the groups, most of them reflecting the specific fatty acyl composition of the diet tested. Testosterone concentration was higher in O and C groups compared with S or G. In agreement with this observation, the activity of both key enzymes involved in testosterone biosynthesis (3-beta-HSD and 17-beta-HSD) was higher in O and C groups with significant differences between them (O > C). A significant negative correlation was found between cellular testosterone production and cellular cholesterol ester content. Additionally, testosterone concentration directly correlated with cholesterol levels. We conclude that dietary oils qualitatively and quantitatively modified the lipid composition of interstitial cells, producing either a direct or indirect regulatory effect on testicular steroidogenic function. PMID- 19130111 TI - Macrobrachium borellii hepatopancreas contains a mitochondrial glycerol-3 phosphate acyltransferase which initiates triacylglycerol biosynthesis. AB - Mammals express four isoforms of glycerol-3-phosphate acyltransferase (GPAT). The mitochondrial isoform GPAT1 may have been the acyltransferase that appeared first in evolution. The hepatopancreas of the crustacean Macrobrachium borellii has a high capacity for triacylglycerol (TAG) biosynthesis and storage. In order to understand the mechanism of glycerolipid biosynthesis in M. borellii, we investigated its hepatopancreas GPAT activity. In hepatopancreas mitochondria, we identified a GPAT activity with characteristics similar to those of mammalian GPAT1. The activity was resistant to inactivation by SH-reactive N ethylmaleimide, it was activated by polymyxin-B, and its preferred substrate was palmitoyl-CoA. The reaction products were similar to those of mammalian GPAT1. A 70-kDa protein band immunoreacted with an anti-rat liver GPAT1 antibody. Surprisingly, we did not detect high GPAT specific activity in hepatopancreas microsomes. GPAT activity in microsomes was consistent with mitochondrial contamination, and its properties were similar to those of the mitochondrial activity. In microsomes, TAG synthesis was not dependent on the presence of glycerol-3 phosphate as a substrate, and the addition of monoacylglycerol as a substrate increased TAG synthesis 2-fold. We conclude that in M. borellii the de novo triacylglycerol biosynthetic pathway can be completed in the mitochondria. In contrast, TAG synthesis in the ER may function via the monoacylglycerol pathway. PMID- 19130112 TI - A theoretical framework for beta-glucan degradation during barley malting. AB - During malting, barley germinates and produces hydrolytic enzymes that de structure the endosperm, making the grains soft and friable. This process starts close to the embryo and spreads throughout the whole grain. It is leaded by the degradation of cell walls, which are mainly constituted of beta-glucans. Fast and extended breakdown of beta-glucans occurs by means of an expanding reaction front driven by beta-glucanase, and appears to follow pseudo-first-order kinetics. Endosperm permeabilization to macromolecules is closely linked to the dismantling of cell walls, thus that access to beta-glucans by beta-glucanase itself is limited. It is shown that the kinetics of beta-glucan degradation during malting are consequent to this condition, and can be explained according to an anomalous evolution of the reverse quasi-steady-state approximation (rQSSA) for enzymatic reactions. In fact, kinetics based on the rQSSA include a transient phase wherein fast substrate depletion is indeed of pseudo-first-order. In the germinating barley, the conditions in which the physical modification of the endosperm occurs are shown to be suitable for the fast transient to persist in dynamic equilibrium while it progressively expands throughout the grain, depleting most beta-glucans and, then, establishing the overall kinetics of beta-glucan breakdown. PMID- 19130113 TI - Biochemical responses and accumulation properties of long-chain perfluorinated compounds (PFOS/PFDA/PFOA) in juvenile chickens (Gallus gallus). AB - One-day-old male chickens were exposed via oral gavage to mixtures of perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), and perfluorodecanoate (PFDA) at either a low dose (0.1 mg/kg body weight [b.w.]) or a high dose (1.0 mg/kg b.w.), or a saline/ethanol vehicle control, three times a week for 3 weeks. After 3 weeks of exposure, half of the chicks were sacrificed and the other half were allowed to depurate for a further 3 weeks. No dose dependent statistically significant differences in body/organ weights were observed among treatment and control groups after 3 weeks of exposure or after three 3 of depuration. Neither 15 histological nor 14 measured plasma biochemical parameters were significantly different in chicks from the exposed groups and vehicle controls. PFOS, PFDA, and PFOA concentrations in blood/liver/kidney samples were measured throughout the exposure and depuration periods at different time intervals. PFOS and PFDA accumulated at much higher concentrations than PFOA during the experimental periods. Interestingly, PFOS and PFDA accumulation patterns in the blood were similar during the exposure and depuration periods. The half-lives for each PFC at the 0.1 and 1.0 mg/kg doses were, respectively, approximately 15 and 17 days for PFOS, 11 and 16 days for PFDA, and 3.9 and 3.9 days for PFOA. PFDA accumulation in organs was greater than or similar to that of PFOS: the liver was the main target during exposure and the blood was the main reservoir during depuration. These results indicate that exposure to a 1.0-mg mixture of PFOS/PFDA/PFOA/kg b.w. has no adverse effect on juvenile chickens. PMID- 19130115 TI - Percutaneous radiologic gastrostomy: a modified Chiba-needle puncture technique with single gastropexy. AB - BACKGROUND: To evaluate the feasibility, safety, and effectiveness of percutaneous radiologic gastrostomy (PRG) with a modified Chiba-needle puncture technique with the use of a single gastropexy in the same puncture tract. METHODS: In 80 patients with dysphagia, the stomach was punctured with a 21-gauge Chiba-needle, which was exchanged for a 6-Fr Neff catheter. Eleven patients with a high-lying stomach and four patients with surgically altered gastric morphology were included. A Cope suture anchor was then deployed into stomach lumens through the Neff catheter to achieve gastropexy. Then, following serial dilation, a 14-Fr diameter locking loop catheter was inserted. RESULTS: All 80 patients successfully underwent PRG. Only a single puncture attempt was required for 78 patients (98%) and two attempts for two patients each. The average procedure time was 5 min 38 s. Three patients (3.8%) had major complications such as hemorrhage, deep stomal infection, and worsening pneumoperitoneum requiring tube removal. Procedure-related mortality and 30-day mortality was 0% and 5%, respectively. CONCLUSIONS: PRG with the modified Chiba-needle puncture technique with use of single gastropexy in the same puncture tract was feasible, safe, and effective, especially in patients with high-lying or surgically altered stomachs. PMID- 19130116 TI - Prostate MR imaging at 3T using a phased-arrayed coil in predicting locally recurrent prostate cancer after radiation therapy: preliminary experience. AB - The purpose of this study was to retrospectively assess the diagnostic performance of diffusion-weighted imaging (DWI) and dynamic contrast-enhanced imaging (DCEI) at 3T in predicting locally recurrent prostate cancer after radiation therapy. Twenty-four patients with a rising prostate-specific antigen level after treatment with radiation therapy underwent prostate MR imaging at 3T, followed by transrectal ultrasound-guided biopsy. MRI findings and biopsy results were correlated in six prostate sectors of both peripheral zones. Two radiologists in consensus reviewed the MR images and rated the likelihood of recurrent cancer on a 5-point scale. Out of the 144 prostate sectors, 37 (26%) sectors were positive for cancer in ten patients. For predicting locally recurrent cancer, the sensitivity and specificity of DWI, DCEI, and combined DCEI and DWI were higher than those for T2-weighted imaging (T2WI). The accuracy of DWI, DCEI and combined DCEI and DWI was greater than that of T2WI. A significantly greater Az was determined for combined DCEI and DWI (Az = 0.863, P < 0.05) as compared with T2WI, DCEI, and DWI. For predicting locally recurrent prostate cancer after radiation therapy, our preliminary results suggest that the use of either DWI or DCEI is superior to the use of T2WI. PMID- 19130117 TI - Whole body imaging in the abdominal cancer patient: pitfalls of PET-CT. AB - Proper interpretation of PET-CT images requires knowledge of the normal physiological distribution of the tracer, frequently encountered physiological variants, and benign pathological causes of FDG uptake that can be confused with a malignant neoplasm. In addition, not all malignant processes are associated with avid tracer uptake. A basic knowledge of the technique of image acquisition is also required to avoid pitfalls such as misregistration of anatomical and scintigraphic data. This article reviews these potential pitfalls as they apply to the abdomen and pelvis of patients with cancer. PMID- 19130118 TI - What is a surgical complication? PMID- 19130119 TI - The increasing use of liver transplantation surgical techniques in the management of renal and nonrenal cancer with or without inferior vena cava tumor thrombus. PMID- 19130120 TI - 3D stereoscopic visualization of fenestrated stent grafts. AB - The purpose of this study was to present a technique of stereoscopic visualization in the evaluation of patients with abdominal aortic aneurysm treated with fenestrated stent grafts compared with conventional 2D visualizations. Two patients with abdominal aortic aneurysm undergoing fenestrated stent grafting were selected for inclusion in the study. Conventional 2D views including axial, multiplanar reformation, maximum-intensity projection, and volume rendering and 3D stereoscopic visualizations were assessed by two experienced reviewers independently with regard to the treatment outcomes of fenestrated repair. Interobserver agreement was assessed with Kendall's W statistic. Multiplanar reformation and maximum-intensity projection visualizations were scored the highest in the evaluation of parameters related to the fenestrated stent grafting, while 3D stereoscopic visualization was scored as valuable in the evaluation of appearance (any distortions) of the fenestrated stent. Volume rendering was found to play a limited role in the follow-up of fenestrated stent grafting. 3D stereoscopic visualization adds additional information that assists endovascular specialists to identify any distortions of the fenestrated stents when compared with 2D visualizations. PMID- 19130121 TI - Combined retrograde-antegrade external iliac artery occlusion recanalization through collaterals: description of a new technique. PMID- 19130122 TI - Dual-energy CT angiography in peripheral arterial occlusive disease. AB - We sought to study the accuracy of dual-energy computed tomographic angiography (DE-CTA) for the assessment of symptomatic peripheral arterial occlusive disease of the lower extremity by using the dual-energy bone removal technique compared with a commercially available conventional bone removal tool. Twenty patients underwent selective digital subtraction angiography and DE-CTA of the pelvis and lower extremities. CTA data were postprocessed with two different applications: conventional bone removal and dual-energy bone removal. All data were reconstructed and evaluated as 3D maximum-intensity projections. Time requirements for reconstruction were documented. Sensitivity, specificity, accuracy, and concordance of DE-CTA regarding degree of stenosis and vessel wall calcification were calculated. A total of 359 vascular segments were analyzed. Compared with digital subtraction angiography, sensitivity, specificity, and accuracy, respectively, of CTA was 97.2%, 94.1%, and 94.7% by the dual-energy bone removal technique. The conventional bone removal tool delivered a sensitivity of 77.1%, a specificity of 70.7%, and an accuracy of 72.0%. Best results for both postprocessing methods were achieved in the vascular segments of the upper leg. In severely calcified segments, sensitivity, specificity, and accuracy stayed above 90% by the dual-energy bone removal technique, whereas the conventional bone removal technique showed a substantial decrease of sensitivity, specificity, and accuracy. DE-CTA is a feasible and accurate diagnostic method in the assessment of symptomatic peripheral arterial occlusive disease. Results obtained by DE-CTA are superior to the conventional bone removal technique and less dependent on vessel wall calcifications. PMID- 19130123 TI - Delayed development of pneumothorax after pulmonary radiofrequency ablation. AB - Acute pneumothorax is a frequent complication after percutaneous pulmonary radiofrequency (RF) ablation. In this study we present three cases showing delayed development of pneumothorax after pulmonary RF ablation in 34 patients. Our purpose is to draw attention to this delayed complication and to propose a possible approach to avoid this major complication. These three cases occurred subsequent to 44 CT-guided pulmonary RF ablation procedures (6.8%) using either internally cooled or multitined expandable RF electrodes. In two patients, the pneumothorax, being initially absent at the end of the intervention, developed without symptoms. One of these patients required chest drain placement 32 h after RF ablation, and in the second patient therapy remained conservative. In the third patient, a slight pneumothorax at the end of the intervention gradually increased and led into tension pneumothorax 5 days after ablation procedure. Underlying bronchopleural fistula along the coagulated former electrode track was diagnosed in two patients. In conclusion, delayed development of pneumothorax after pulmonary RF ablation can occur and is probably due to underlying bronchopleural fistula, potentially leading to tension pneumothorax. Patients and interventionalists should be prepared for delayed onset of this complication, and extensive track ablation following pulmonary RF ablation should be avoided. PMID- 19130124 TI - Magnetic resonance angiography: current status in the planning and follow-up of endovascular treatment in lower-limb arterial disease. AB - Magnetic resonance angiography (MRA) has become an established imaging modality in the management of lower-limb arterial disease, with emerging roles in treatment planning and follow-up. Contrast-enhanced MRA is now the most widely used technique with clinically acceptable results in the majority of patients. Difficulties in imaging and image interpretation are recognised in certain subgroups, including patients with critical limb ischaemia as well as patients with stents. Although newer contrast agents and refined imaging protocols may offer some solutions to these problems, this optimism is balanced by concerns about the toxicity of certain gadolinium chelates. Further development of interventional MRA remains one of the most significant challenges in the development of magnetic resonance imaging-guided peripheral vascular intervention. The status of MRA in managing patients with lower-limb arterial disease in current clinical practice is reviewed. PMID- 19130125 TI - Exploring the antagonistic producer Streptomyces MSI051: implications of polyketide synthase gene type II and a ubiquitous defense enzyme phospholipase A2 in the host sponge Dendrilla nigra. AB - To explore the functional role of the antagonistic producer strain Streptomyces dendra sp. nov. MSI051 in the host sponge Dendrilla nigra, hypothetical factors including the antagonistic potential of MSI051 against biofilm bacteria and a ubiquitous defense enzyme phospholipase A2 (PLA2) in host sponge as well as in bacterial symbiont MSI051 were determined. The host sponge D. nigra and associated bacterial symbiont MSI051 contained high levels of PLA2. The host sponge showed PLA2 activity to the extent of 1032 U/L, with a specific activity of 2021 U/g, and strain MSI051 showed similar activity. The findings of the present study suggest that PLA2 in the sponge-associated bacteria might have an integrated functional role in the host defense system of marine sponges. This report may be the first on the role of PLA2 activity in sponge-associated bacteria. Isolate MSI051 was a potential antagonistic producer which showed a broad spectrum of antibacterial activity. Polyketide synthase gene type II in MSI051 ultimately evidenced the antagonistic potential. Antimicrobial activity was found to be positively skewed toward biofilm bacteria. This implies a functional role of MSI051 in the protection of host sponge against fouling processes. PMID- 19130126 TI - A mouse model for characterization of gastrointestinal colonization rates among environmental Aeromonas isolates. AB - The colonization rates of 10 different environmental Aeromonas isolates were determined using a novel mouse-streptomycin pretreatment method. As demonstrated, alterations to the colon flora of mice pretreated with streptomycin allowed transient colonization by bacterial species normally excluded by host competition. A novel procedure is described for determining the colonization abilities of Aeromonas isolates under these conditions. The colonization rates of A. salmonicida, A. encheleia, and A. allosaccharophila were either negative or occurred randomly at low levels with respect to concentrations of the dosage consumed by the animals. In contrast, A. hydrophila, A. veronii biovar sobria, and A. caviae exhibited relatively high rates of mouse colon tissue colonization. PMID- 19130127 TI - Characterization of two novel cry8 genes from Bacillus thuringiensis strain BT185. AB - Two novel cry8-type genes, cry8Ea1 and cry8Fa1, obtained from a Holotrichia parallela-specific Bacillus thuringiensis strain, BT185, were characterized. Findings showed that cry8Ea1 and cry8Fa1 encoded polypeptides of 1164 and 1174 amino acid residues, respectively. The deduced amino acid sequences of both Cry8Ea1 and Cry8Fa1 polypeptides are the most similar to that of Cry8Ba1. Eight conserved blocks (blocks 1-8) exist in Cry8Ea1 and Cry8Fa1 polypeptides compared with known Cry proteins. Cry8Ea1 and the Cry8Fa1 toxins could form spheric crystals when they were expressed in the acrystalliferous mutant strain HD73(-). The spores and crystals from the recombinant strain containing cry8Ea1 were toxic to Holotrichia parallela, with an LC(50) of 0.0875 x 10(8) colony-forming units (CFU)/g. However, Cry8Fa1 expressed in the recombinant strain was not toxic to H. parallela, Anomala corpulenta, or H. oblita. PMID- 19130128 TI - A new type of DNA polymorphism identified in the species-specific DNA region originating from the Candida albicans mitochondrial genome. AB - Mitochondrial DNA of the pathogenic yeast Candida albicans contains a species specific DNA region, designated EO3, within the duplicated region in its stem-and loop structure, as reported in our previous paper. We have revealed that the EO3 region exhibits a DNA-size polymorphism, yielding three fragment length types: L, M, and S. This polymorphism is due to the presence (or absence) of two small nucleotide sequences of ~50 bp (designated a and b) within the EO3 region: type L possesses both a and b, type M possesses a alone, and type S possesses neither a nor b. Here we have identified a new type of EO3, which possesses b but not a, designated type M-II. Restriction fragment analyses with BglII revealed accurate discrimination of EO3 into four types: L, M-I, M-II, and S. Moreover, we show that these small nucleotide segments a and b were encompassed by the characteristic nucleotide sequence with a common inverted repeat structure. In the present report we propose a revised version of EO3 typing, suggesting epidemiological usefulness as an alternative tool for species-specific detection of C. albicans, and discuss the potentiality of EO3 for generating more variations of DNA polymorphism. PMID- 19130129 TI - Canine RD3 mutation establishes rod-cone dysplasia type 2 (rcd2) as ortholog of human and murine rd3. AB - Rod-cone dysplasia type 2 (rcd2) is an autosomal recessive disorder that segregates in collie dogs. Linkage disequilibrium and meiotic linkage mapping were combined to take advantage of population structure within this breed and to fine map rcd2 to a 230-kb candidate region that included the gene C1orf36 responsible for human and murine rd3, and within which all affected dogs were homozygous for one haplotype. In one of three identified canine retinal RD3 splice variants, an insertion was found that cosegregates with rcd2 and is predicted to alter the last 61 codons of the normal open reading frame and further extend the open reading frame. Thus, combined meiotic linkage and LD mapping within a single canine breed can yield critical reduction of the disease interval when appropriate advantage is taken of within-breed population structure. This should permit a similar approach to tackle other hereditary traits that segregate in single closed populations. PMID- 19130130 TI - Neuroplasticity and swallowing. AB - Recent research has suggested that the central nervous system controlling swallowing can undergo experience-dependent plasticity. Moreover, swallowing neuroplastic change may be associated with behavioural modulation. This article presents research evidence suggesting that nonbehavioural and behavioural interventions, as well as injury, can induce swallowing neuroplasticity. These studies indicate that while swallowing and limb neuroplasticity share certain features, certain principles of swallowing neuroplasticity may be distinct. Thus, an understanding of swallowing neuroplasticity is necessary in terms of explaining and predicting the (1) behavioural effects of injury to the swallowing nervous system and (2) effects of swallowing interventions applied in rehabilitation. PMID- 19130132 TI - A motion compounding technique for speckle reduction in ultrasound images. AB - The quality of ultrasound images is usually influenced by speckle noise and the temporal decorrelation of the speckle patterns. To reduce the speckle noise, compounding techniques have been widely applied. Partially correlated images scanned on the same subject cross-section are combined to generate a compound image with improved image quality. However, the compounding technique might introduce image blurring if the transducer or the target moves too fast. This blurring effect becomes especially critical when assessing tissue deformation in clinical motion examinations. In this paper, an ultrasound motion compounding system is proposed to improve the quality of ultrasound motion sequences. The proposed motion compounding technique uses a hierarchical adaptive feature weighted motion estimation method to realign the frames before compounding. Each frame is first registered and warped to the reference frame before being compounded to reduce the speckle noise. Experimental results showed that the motion could be assessed accurately and better visualization could be achieved for the compound images, with improved signal-to-noise and contrast-to-noise ratios. PMID- 19130133 TI - Surgical treatments and prognoses of patients with combined hepatocellular carcinoma and cholangiocarcinoma. AB - BACKGROUND: Combined hepatocellular carcinoma and cholangiocarcinoma is a very rare form of primary liver cancer containing components of both tumor types. We evaluated the effectiveness of surgical treatment and factors related to survival and recurrence. PATIENTS AND METHODS: Of the 2427 patients who underwent hepatectomy or liver transplantation because of a primary hepatic malignancy from January 1989 to July 2006 at the Asan Medical Center, Seoul, Korea, 29 had hepatocellular carcinoma and cholangiocarcinoma as a single mixed or transitional tumor. Their medical records were retrospectively reviewed. RESULTS: Disease-free survival rates at 6 months, 1 year, and 3 years were 51.1%, 38.3%, and 25.6%, respectively. Univariate analysis showed that CA 19-9 above 37 U/ml was predictive of low overall survival (P= .03) and that TNM stage was significantly associated with disease-free survival (P= .04). CONCLUSIONS: Patients with combined hepatocellular carcinoma and cholangiocarcinoma had poor postoperative survival rates. High CA 19-9 level was associated with poorer survival, suggesting that the cholangiocarcinoma portion may be a major determining factor for patient prognosis. Aggressive surgical treatment, including lymph node dissection, may improve survival in patients suspected of or diagnosed with these tumors. PMID- 19130134 TI - Evaluation of 47 soft tissue sarcoma resection specimens after isolated limb perfusion with TNF-alpha and melphalan: histologically characterized improved margins correlate with absence of recurrences. AB - BACKGROUND: Isolated limb perfusion (TM-ILP) is an effective limb-sparing treatment for primarily nonresectable soft tissue sarcomas (STS). Surgical margins of STS after ILP were yet not systematically studied. METHODS: In 47 patients with nonresectable STS, TM-ILP with subsequent tumor resection was performed. Surgical margins were systematically analyzed by light microscopy using the TNM and the Enneking classification. Furthermore, margins were analyzed for tumor regression in terms of improved resectability. Results were correlated with clinical and pathological parameters. RESULTS: Of 47 STS, 44 were classified as high-grade (93.6%) with a median tumor size of 10.0 cm. Primary limb-salvage rate was 85.1%. According to TNM resection margins were complete in 70.2% (R0) and incomplete in 29.8% (R1=21.3%, R2=8.5%). According to Enneking, 27.7% intralesional, 42.6% marginal, 21.3% wide, 2.1% radical, and 6.4% unclassifiable margins were found. Prior surgery and/or radiotherapy significantly decreased margin quality. Ten patients with incomplete resection (three intralesional, seven marginal) had no viable tumor at the plane of dissection, which was designated as "improved margins." Whereas those patients remained relapse free, five patients with viable tumor (not improved margins) at the resection margin had local recurrences. Poor margins were associated with local and distant recurrences and limited disease-specific survival. CONCLUSION: TM-ILP is effective for achieving limb salvage. Histopathology of surgical margins demonstrates cases with so-called "improved margins" after TM-ILP, which are related to a better outcome even in intralesionally resected tumors. Improvement of margins should be further evaluated as a potential relevant prognostic parameter. PMID- 19130135 TI - 5,000 parathyroid operations without frozen section or PTH assays: measuring individual parathyroid gland hormone production in real time. AB - BACKGROUND: Determining the physiologic activity (hormone production) of individual parathyroid glands can provide tremendous guidance during parathyroidectomy. METHODS: A 6.5-year prospective study of 5,000 patients with sporadic, non-multiple endocrime neoplasia (MEN) primary hyperparathyroidism who underwent surgery without frozen section or parathyroid hormone (PTH) assays was conducted. Patients who had a frozen section were not included; the removal of parathyroid glands was determined solely by the physiologic activity of each gland as determined by contained radioactivity. All operations were within 2.25 hours of sestamibi scanning. Ex vivo measurements of parathyroid glands, thyroid nodules, lymph nodes, thymus, and fat were obtained from all patients, constituting >32,000 specimens. All patients had at least two parathyroid glands evaluated; 59% had four glands evaluated. Ratios were compared with histology and preoperative/postoperative labs. RESULTS: Parathyroid glands occur in three distinct groups according to their hormone production, indicating the type of pathology present. Adenomas (n=5,120) contained 57+/-38% of background radioactivity; hyperplastic glands (n=640) contained 16+/-4%; and normal glands (n=9,400) contained 4+/-0.1% (all p<0.00001). Fat and lymph nodes were always less than normal glands (p<0.005). There was no overlap between different tissue types in any individual (p<0.001). Contained radioactivity was a better predictor of cure than histology (p<0.0001). The average operative time was 19.4 minutes with (99.9%) discharged within 5 hours. The initial cure rate was 99.23% with missed contralateral second adenomas causing all failures (subsequently cured). CONCLUSIONS: Measures of sequestered radioactivity is an extremely accurate estimate of individual parathyroid gland hormone production allowing near 100% distinction between normal, hyperplasic, and adenomatous glands as well as distinguishing parathyroids from other neck tissues (fat, lymph nodes, thyroid). This instantaneous measurement is sufficient to determine which glands should be removed and which should remain in situ, while eliminating frozen sections and PTH assays in nearly all patients undergoing parathyroid surgery. This insight allows the operation to progress very rapidly. PMID- 19130136 TI - Liver cell adenoma: a multicenter analysis of risk factors for rupture and malignancy. AB - BACKGROUND: Liver cell adenoma (LCA) is a benign hepatic tumor with poorly characterized risk for spontaneous rupture and malignant transformation. METHODS: Records from five tertiary hepatobiliary centers were reviewed for all patients treated for LCA from 1997 to 2006. Clinicopathological data were collected and analyzed, and factors that were associated with rupture and/or malignant transformation were assessed by using multivariable logistic regression. RESULTS: A total of 124 patients were analyzed, of which 8 (6.5%) were men; 119 patients underwent resection, and 5 patients had embolic therapy only. Mean patient age was 39+/-11 years, and 55% had history of hormone use. Rupture occurred in 31 (25%) cases. Ruptured tumors were larger (10.5+/-4.5 cm vs. 7.2+/-4.8 cm; p=0.001), and no tumor <5 cm ruptured. Patients with ruptured LCAs were more likely to require preoperative blood transfusion (32% vs. 9%, p=0.006), preoperative embolization (16% vs. 1%, p=0.021), and major (>or=3 segments) hepatic resection (65% vs. 32%, p=0.003). By multivariate analysis, increasing tumor size (odds ratio (OR), 7.8; 95% confidence interval (CI), 2.2-26.3; p<0.01) and recent (within 6 months) hormone use (OR, 4.5; 95% CI, 1.5-13.3; p<0.01) remained independently associated with risk of rupture. Five cases (4%) had evidence of underlying malignancy, but none had LCA <8 cm in diameter. CONCLUSION: In this multicenter analysis of patients with LCAs, risk of rupture correlated with increasing tumor size and recent hormone use. Rupture is associated with greater need for preoperative blood transfusion and major hepatic resection. These data suggest that patients with asymptomatic LCAs approaching 4 cm and those requiring hormonal therapy should undergo surgical therapy. PMID- 19130137 TI - Solitary dermal melanoma: beginning or end of the metastatic process? AB - BACKGROUND: Solitary dermal melanoma (SDM) is confined to the dermal and/or subcutaneous tissue without an epidermal component. It is unclear whether this lesion is a subtype of primary melanoma or distant cutaneous metastasis from an unknown primary. We evaluated our large experience to determine the prognosis and optimal management of SDM. METHODS: Our melanoma referral center's database of prospectively acquired records was used for identification and clinicopathologic analysis of patients presenting with SDM between 1971 and 2005. RESULTS: Of 12,817 database patients seen during a 34-year period, 101 (0.8%) had SDM. Of 92 patients free of distant metastasis on initial presentation, 55 (60%) were observed and 37 (40%) underwent surgical nodal staging: regional metastases were identified in 7 (19%). Nodal recurrence occurred in 1 of 30 patients (3.3%) with histopathology-negative nodes compared with 13 of 55 patients (24%) who underwent nodal observation instead of nodal staging. Thus, 21 of 92 patients (23%) had nodal metastasis identified during surgical nodal staging or postoperative nodal observation. At a median follow-up of 68 months, estimated 5-year overall survival rate was 73% for 71 patients with localized disease versus 67% for 21 patients with regional disease (P=0.25) versus 22% for 9 patients with distant disease (P=0.009, regional versus distant disease). CONCLUSIONS: SDM resembles intermediate-thickness primary cutaneous melanoma with respect to prognostic characteristics and clinical evolution, but its rate of distant metastasis justifies radiographic staging and its high rate of regional node metastasis justifies wide excision and sentinel node biopsy. PMID- 19130138 TI - Long-term survival and disease recurrence following portal vein embolisation prior to major hepatectomy for colorectal metastases. AB - BACKGROUND: Portal vein embolisation (PVE) can be used to increase the remnant liver parenchyma volume before major hepatectomy but may stimulate tumour growth. The pattern of disease recurrence and long-term survival has not been adequately addressed. METHODS: Over a period of 7 years 36 patients underwent preoperative PVE before resection of four or more liver segments for colorectal cancer (CRC) liver metastases. PVE was performed when the future liver remnant (FLR) assessed by magnetic resonance imaging (MRI) scan volumetry was less than 30%. Disease free and overall survival was compared with a control group (65 patients) undergoing extended right/right hepatectomy for CRC metastases without PVE during the same time period. RESULTS: PVE was successful in all patients. PVE increased the median FLR volume by 37% [295 ml (22%) to 404 ml (32%), p < 0.0001]. 61% of patients undergoing PVE proceeded to liver resection (n = 22). Twelve patients (33%) developed disease progression following PVE. The 5-year survival after liver resection with PVE was 25%, compared with 50% without PVE. The 5-year disease-free survival was 30% post PVE and 50% without PVE. CONCLUSION: We conclude that PVE significantly increases the future liver remnant. Only two thirds of patients proceed to resection because of disease progression. Long-term survival is less than in patients who do not require PVE. The effect of PVE on tumour growth requires investigation. PMID- 19130139 TI - Blood neutrophil-to-lymphocyte ratio predicts survival in patients with colorectal liver metastases treated with systemic chemotherapy. AB - BACKGROUND: Whether neutrophil-to-lymphocyte ratio (NLR) predicts survival of patients with colorectal liver metastases (CLM) treated with systemic chemotherapy remains unclear. METHODS: Clinicopathologic data were reviewed for patients with CLM treated with chemotherapy and resection (n=200) or chemotherapy only (n=90). Univariate and multivariate analyses for prognostic factors were performed. In the resection group, whether chemotherapy normalizes high NLR and the effect of NLR normalization on survival were evaluated. RESULTS: In the resection group, patients with preoperative NLR>5 had a worse 5-year survival rate than patients with NLR 5 was the only independent preoperative predictor of worse survival (P=0.016; hazard ratio [HR]=2.22; 95% confidence interval [95% CI], 1.16-4.25). In the nonresection group, patients with prechemotherapy NLR>5 had a worse 3-year survival rate than patients with NLR 5 was the only independent predictor of worse survival (P=0.001; HR = 2.91; 95% CI, 1.54-5.50). In the resection group, chemotherapy normalized high NLR in 17 of 25 patients, and these 17 patients had better survival than the 8 patients with high NLR both before chemotherapy and before surgery (P=0.021). CONCLUSION: NLR independently predicts survival in patients with CLM treated with chemotherapy followed by resection or chemotherapy only. When chemotherapy normalizes high NLR, improved survival is expected. PMID- 19130140 TI - Meta-analysis of the clamp-crushing technique for transection of the parenchyma in elective hepatic resection: back to where we started? AB - BACKGROUND: After introduction of the clamp-crushing technique in the 1970s, various devices have been developed for transection of the liver with the aim of further reducing blood loss and improving the outcome of patients who undergo hepatic resection. We performed a meta-analysis to quantitatively compare the clamp-crushing technique to any subsequently introduced transection technique with respect to patients' perioperative outcome. METHODS: A systematic literature search was conducted to identify randomized controlled trials comparing the clamp crushing technique to any alternative method of hepatic transection. Relative risks (RR) were calculated for each outcome and reported along with their 95% confidence intervals (95% CI). Meta-analyses were stratified for the various types of transection techniques compared with the clamp-crushing technique and were carried out by a random effects model. RESULTS: Seven randomized controlled trials with a total of 554 patients were included in final analyses. Analyses of overall morbidity (RR .89; 95% CI, .63-1.25), biliary leakage (RR 1.03; 95% CI, .50-2.13), transfusion rates (RR .69; 95% CI, .31-1.51), and mortality RR (.20; 95% CI, .02-1.65) revealed no difference between the clamp-crushing and alternative transection techniques. None of the identified studies demonstrated a clinically important benefit of an alternative transection method in terms of blood loss, parenchymal injury, transection time, and hospital stay. CONCLUSIONS: This meta-analysis does not indicate a benefit of any alternative transection technique on patients' perioperative outcome compared with the clamp-crushing technique. The clamp-crushing technique remains the reference technique for transection of the parenchyma in elective hepatic resection. PMID- 19130141 TI - In vitro chemoresistance testing in well-differentiated carcinoid tumors. AB - BACKGROUND: Well-differentiated, "typical" carcinoid tumors traditionally have a very poor response to chemotherapy. We hypothesized that tumor specimens from well-differentiated carcinoid tumors would be highly resistant to the effects of chemotherapy when tested against a variety of antineoplastic agents in vitro. METHODS: Ninety-eight typical carcinoid specimens were surgically harvested, cultured, and tested against antineoplastics in vitro. (3)H-Thymidine incorporation was used to assess the percentage of cell-growth inhibition (PCI) of tested specimens. PCI was used to determine if specimens had extreme drug resistance (EDR), intermediate drug resistance (IDR), or low drug resistance (LDR) to each reagent against which they were tested. RESULTS: Seventy specimens generated results. Each was tested with an average of six drugs. The mean proportions of drugs classified as LDR, IDR, and EDR were 0.48 (range 0-1), 0.34 (range 0-1), and 0.18 (range 0-0.80), respectively. The mean numbers of drugs per specimen exhibiting LDR, IDR, and EDR chemoresistance were 2.7, 2.1, and 1.2, respectively. 57 of 70 specimens (81%) had LDR to at least two drugs. 5-FU had the highest frequency of low chemoresistance at 69%, followed by doxorubicin at 67%. Low in vitro resistance to chemotherapeutics was prevalent among typical carcinoids, while EDR was comparatively infrequent. CONCLUSIONS: This implies that there may be less clinical chemoresistance and more chemosensitivity among typical carcinoid tumors than clinical trials have previously revealed. These findings warrant additional investigations assessing the response of carcinoid tumors to assay-guided chemotherapy regimens. PMID- 19130142 TI - PET imaging studies in rhesus monkey with the cannabinoid-1 (CB1) receptor ligand [11C]CB-119. AB - PURPOSE: The in vitro and in vivo evaluation of the selective, high affinity (human CB1 IC(50) 0.49 nM) inverse agonist CB1R tracer [(11)C]CB-119, a close analog of the previously disclosed [(18)F]MK-9470, was undertaken. PROCEDURES: [(11)C]CB-119 was synthesized with high specific activity by alkylation of a phenolic precursor with [(11)C]methyl iodide. In vitro autoradiographic studies using rhesus brain slices were carried out using [(3)H]CB-119, and in vivo imaging studies were carried out using [(11)C]CB-119 in rhesus monkeys under baseline and blocked conditions. RESULTS: Autoradiographic studies in rhesus brain showed the expected distribution pattern for CB1R with highest binding in the cerebral cortex, cerebellum, caudate/putamen, globus pallidus, substantia nigra, and hippocampus. Lower binding was seen in the posterior hypothalamus, ventral tegmental area, and periventricular gray area, and the lowest binding was in the thalamic nuclei. The binding of [(3)H]CB-119 was fully blocked by the addition of 10 microM CB-119. Rhesus positron emission tomography imaging studies showed very good brain uptake and a distribution pattern consistent with that seen in the autoradiographic studies. The kinetics of tracer uptake was slow. The brain uptake was blocked by pretreatment with taranabant, a CB1R inverse agonist. The specific signal (total/nonspecific) in rhesus putamen at 90 min was approximately 6:1. CONCLUSIONS: [(11)C]CB-119 is a suitable tracer for imaging central CB1 receptors. PMID- 19130144 TI - The applicability of ordinary least squares to consistently short distances between taxa in phylogenetic tree construction and the normal distribution test consequences. AB - Short phylogenetic distances between taxa occur, for example, in studies on ribosomal RNA-genes with slow substitution rates. For consistently short distances, it is proved that in the completely singular limit of the covariance matrix ordinary least squares (OLS) estimates are minimum variance or best linear unbiased (BLU) estimates of phylogenetic tree branch lengths. Although OLS estimates are in this situation equal to generalized least squares (GLS) estimates, the GLS chi-square likelihood ratio test will be inapplicable as it is associated with zero degrees of freedom. Consequently, an OLS normal distribution test or an analogous bootstrap approach will provide optimal branch length tests of significance for consistently short phylogenetic distances. As the asymptotic covariances between branch lengths will be equal to zero, it follows that the product rule can be used in tree evaluation to calculate an approximate simultaneous confidence probability that all interior branches are positive. PMID- 19130143 TI - A molecular imaging paradigm to rapidly profile response to angiogenesis-directed therapy in small animals. AB - PURPOSE: The development of novel angiogenesis-directed therapeutics is hampered by the lack of non-invasive imaging metrics capable of assessing treatment response. We report the development and validation of a novel molecular imaging paradigm to rapidly assess response to angiogenesis-directed therapeutics in preclinical animal models. PROCEDURES: A monoclonal antibody-based optical imaging probe targeting vascular endothelial growth factor receptor-2 (VEGFR2) expression was synthesized and evaluated in vitro and in vivo via multispectral fluorescence imaging. RESULTS: The optical imaging agent demonstrated specificity for the target receptor in cultured endothelial cells and in vivo. The agent exhibited significant accumulation within 4T1 xenograft tumors. Mice bearing 4T1 xenografts and treated with sunitinib exhibited both tumor growth arrest and decreased accumulation of NIR800-alphaVEGFR2ab compared to untreated cohorts (p = 0.0021). CONCLUSIONS: Molecular imaging of VEGFR2 expression is a promising non invasive biomarker for assessing angiogenesis and evaluating the efficacy of angiogenesis-directed therapies. PMID- 19130145 TI - A traveling wave model for invasion by precursor and differentiated cells. AB - We develop and investigate a continuum model for invasion of a domain by cells that migrate, proliferate and differentiate. The model is applicable to neural crest cell invasion in the developing enteric nervous system, but is presented in general terms and is of broader applicability. Two cell populations are identified and modeled explicitly; a population of precursor cells that migrate and proliferate, and a population of differentiated cells derived from the precursors which have impaired migration and proliferation. The equation describing the precursor cells is based on Fisher's equation with the addition of a carrying-capacity limited differentiation term. Two variations of the proliferation term are considered and compared. For most parameter values, the model admits a traveling wave solution for each population, both traveling at the same speed. The traveling wave solutions are investigated using perturbation analysis, phase plane methods, and numerical techniques. Analytical and numerical results suggest the existence of two wavespeed selection regimes. Regions of the parameter space are characterized according to existence, shape, and speed of traveling wave solutions. Our observations may be used in conjunction with experimental results to identify key parameters determining the invasion speed for a particular biological system. Furthermore, our results may assist experimentalists in identifying the resource that is limiting proliferation of precursor cells. PMID- 19130146 TI - Osteoid osteoma of the scaphoid presenting with radiocarpal arthritis: a case report. AB - A case of osteoid osteoma of the scaphoid presenting as painful monoarticular arthritis is presented. Degenerative arthritis, associated with osteoid osteoma of the carpus, has not been described. The implications for treatment are discussed. PMID- 19130147 TI - Challenges in the management of positive fecal occult blood tests. AB - BACKGROUND: Many patients with a positive fecal occult blood test (FOBT) do not undergo follow-up evaluations. OBJECTIVE: To identify the rate of follow-up colonoscopy following a positive FOBT and determine underlying reasons for lack of follow-up. DESIGN: It is a retrospective chart review. PARTICIPANTS: The subject group consisted of 1,041 adults with positive FOBTs within a large physician group practice from 2004 to 2006. MEASUREMENTS: We collected data on reasons for ordering FOBT, presence of prior colonoscopy, completed evaluations, and results of follow-up tests. We fit a multivariable logistic regression model to identify predictors of undergoing follow-up colonoscopy. RESULTS: Most positive FOBTs were ordered for routine colorectal cancer screening (76%), or evaluation of anemia (13%) or rectal bleeding (7%). Colonoscopy was completed in 62% of cases, with one-third of these procedures identifying a colorectal adenoma (29%) or cancer (4%). Factors associated with higher rates of follow-up colonoscopy included obtaining the FOBT for routine colorectal screening (odds ratio (OR) 1.59, 95% confidence interval (CI) 1.11-2.29) and consultation with gastroenterology (OR 1.99, 95% CI 1.46-2.72). Patients were less likely to undergo colonoscopy if they were older than 80 years old (OR 0.54, 95% CI 0.31 0.92), younger than 50 years old (OR 0.44, 95% CI 0.28-0.70), uninsured (OR 0.50, 95% CI 0.27-0.93), or had undergone colonoscopy within the prior five years (OR 0.32, 95% CI 0.23-0.44). CONCLUSIONS: Clinical decisions and patient factors available at the time of ordering an FOBT impact performance of colonoscopy. Targeting physicians' understanding of the use of this test may improve follow-up and reduce inappropriate use of this test. PMID- 19130148 TI - Primary care physicians' use of an electronic medical record system: a cognitive task analysis. AB - OBJECTIVE: To describe physicians' patterns of using an Electronic Medical Record (EMR) system; to reveal the underlying cognitive elements involved in EMR use, possible resulting errors, and influences on patient-doctor communication; to gain insight into the role of expertise in incorporating EMRs into clinical practice in general and communicative behavior in particular. DESIGN: Cognitive task analysis using semi-structured interviews and field observations. PARTICIPANTS: Twenty-five primary care physicians from the northern district of the largest health maintenance organization (HMO) in Israel. RESULTS: The comprehensiveness, organization, and readability of data in the EMR system reduced physicians' need to recall information from memory and the difficulty of reading handwriting. Physicians perceived EMR use as reducing the cognitive load associated with clinical tasks. Automaticity of EMR use contributed to efficiency, but sometimes resulted in errors, such as the selection of incorrect medication or the input of data into the wrong patient's chart. EMR use interfered with patient-doctor communication. The main strategy for overcoming this problem involved separating EMR use from time spent communicating with patients. Computer mastery and enhanced physicians' communication skills also helped. CONCLUSIONS: There is a fine balance between the benefits and risks of EMR use. Automaticity, especially in combination with interruptions, emerged as the main cognitive factor contributing to errors. EMR use had a negative influence on communication, a problem that can be partially addressed by improving the spatial organization of physicians' offices and by enhancing physicians' computer and communication skills. PMID- 19130150 TI - Defining burnout as a dichotomous variable. PMID- 19130149 TI - Physician job satisfaction and quality of care among hospital employed physicians in Japan. AB - BACKGROUND: Physician job satisfaction is reportedly associated with interpersonal quality of care, such as patient satisfaction, but its association with technical quality of care, as determined by whether patients are offered recommended services, is unknown. OBJECTIVE: We explored whether the job satisfaction of hospital-employed physicians in Japan is associated with the technical quality of care, with an emphasis on process qualities as measured by quality indicators. DESIGN: Cross-sectional study linking data from physician surveys with data abstracted from outpatient charts. PARTICIPANTS: A total of 53 physicians working at 13 hospitals in Japan participated. Medical records covering 568 patients were reviewed. MEASUREMENTS: Disease-specific indicators related to the care of patients with hypertension, type 2 diabetes, and asthma, as well as disease-independent measures of the process of care were abstracted. We analyzed the association between the quality of care score for individual physicians, which is defined as the percentage of quality indicators satisfied among the total for which their patients were eligible, and physician job satisfaction, which was measured by a validated scale. RESULTS: No statistically significant association between physician job satisfaction and quality of care was observed. A 1-standard deviation (SD) increment in the physician job satisfaction scale was associated with an increase of only 0.3% for overall quality (P = 0.85), -3.0% for hypertension (P = 0.22), 2.5% for type 2 diabetes (P = 0.44), 8.0% for asthma (P = 0.21), and -0.4% for cross-cutting care (P = 0.76). CONCLUSION: Contrary to the positive association reported between physician job satisfaction and high quality of interpersonal care, no association was seen between physician job satisfaction and the technical quality of care. PMID- 19130151 TI - Systematic review of minimally invasive pancreatic resection. AB - BACKGROUND: Pancreatic resection is associated with a significant morbidity. Efforts to reduce hospital stay and enhance recovery have seen the introduction of minimally invasive surgical techniques. This article reviews the current published literature on the safety and efficacy of minimally invasive surgery of the pancreas. METHODS: An electronic search of the PubMed and Embase databases was performed from 1996 to May 2008 to identify all relevant publications; studies meeting predefined inclusion criteria were retrieved and analyzed using a standardized protocol. Data on the safety and efficacy of minimally invasive surgery of the pancreas were recorded and analyzed. RESULTS: Of 565 abstracts reviewed, 39 studies were identified as eligible for inclusion. There were 37 case series and two case control studies. Compared with open pancreatic surgery, minimally invasive pancreatic resection is similar in terms of morbidity and mortality. Blood loss and length of stay are decreased. CONCLUSIONS: Laparoscopic distal pancreatic resection and enucleation of insulinoma appear to be safe procedures with reduced hospital stay, though morbidity remains significant. The evidence for laparoscopic pancreaticoduodenectomy is in its infancy, but the authors feel it is unlikely that many centers will achieve sufficient case load to make the introduction of minimally invasive resection feasible. PMID- 19130152 TI - Intestinal afferent nerve sensitivity is increased during the initial development of postoperative ileus in mice. AB - INTRODUCTION: Neuronal reflex inhibition of gastrointestinal motility is a key mechanism in the development of postoperative ileus (POI). The aim of our study was to determine whether intestinal afferent nerve fibers are sensitized during the first hours after surgery contributing to this mechanism. METHODS: Under enflurane anesthesia, C57BL/6 mice underwent laparotomy followed by sham treatment or standardized small bowel manipulation to induce POI. After 1, 3, or 9 h, extracellular multi-unit mesenteric afferent nerve recordings were performed in vitro from 2 cm segments of jejunum (subgroups n = 6) superfused with Kreb's buffer (32 degrees C, gassed with O(2)/CO(2) mixture). Segments were cannulated to monitor luminal pressure and intestinal motility. Afferent impulses as response to bradykinin (0.5 microM) and to mechanical ramp distension of the intestinal lumen from 0 to 80 cmH(2)O were recorded. RESULTS: At 1 h, amplitudes of intestinal contractions were 0.8 +/- 0.2 cmH(2)O after induction of POI and 5.0 +/- 0.8 cmH(2)O in sham controls (mean +/- SEM; p < 0.01). A similar difference was observed for segments harvested at 3 and 9 h. Afferent firing to serosal bradykinin was increased at 1, 3, and 9 h in POI segments compared to sham controls (p < 0.05 at 1 h, p < 0.01 at 3 and 9 h). During distension with high pressures, afferent firing rate was increased at 1 and 3 h in segments after induction of POI compared to sham controls. Nine hours postoperatively, contracted and dilated segments were observed during POI that were investigated separately. While afferent firing in dilated segments was increased to 176 +/- 16 imp s(-1) at 80 cmH(2)O luminal distension (p < 0.01), it was 46 +/- 5 imp s(-1) in contracted segments (p < 0.001) compared to 77 +/- 4 imp s(-1) in sham controls. CONCLUSIONS: Afferent firing to bradykinin and high threshold distension is augmented in the early phase of POI. As these stimuli are known to sensitize predominantly spinal afferents, this mechanism may contribute to reflex inhibition of intestinal motility during POI. PMID- 19130153 TI - The incidental asymptomatic pancreatic lesion: nuisance or threat? AB - INTRODUCTION: Although asymptomatic pancreatic lesions (APLs) are being discovered incidentally with increasing frequency, their true significance remains uncertain. Treatment decisions pivot off concerns for malignancy but at times might be excessive. To understand better the role of surgery, we scrutinized a spectrum of APLs as they presented to our surgical practice over defined periods. METHODS: All incidentally identified APLs that were operated upon during the past 5 years were clinically and pathologically annotated. Among features evaluated were method/reason for detection, location, morphology, interventions, and pathology. For the past 2 years, since our adoption of the Sendai guidelines for cystic lesions, we scrutinized our approach to all patients presenting with APLs, operated upon or not. RESULTS: Over 5 years, APLs were identified most frequently during evaluation of: genitourinary/renal (16%), asymptomatic rise in liver function tests (LFTs; 13%), screening/surveillance (7%), and chest pain (6%). APLs occurred throughout the pancreas (body/tail 63%; head/uncinate 37%) with 48% being solid. One hundred ten operations were performed with no operative mortality including 89 resections (distal 57; Whipple 32) and 21 other procedures. Morbidity was equivalent or better than those cases performed for symptomatic lesions during the same time frame. During these 5 years, APLs accounted for 23% of all pancreatic resections we performed. In all, 22 different diagnoses emerged including non-malignant intraductal papillary mucinous neoplasm (IPMN; 17%), serous cystadenoma (14%), and neuroendocrine tumors (13%), while 6% of patients had >1 distinct pathology and 12% had no actual pancreatic lesion at all. Invasive malignancy was present 17% of the time, while carcinoma in situ or metastases was identified in an additional eight patients. Thus, the overall malignancy rate for APLs equals 24% and these patients were substantially older (68 vs 58 years; p = 0.003). An asymptomatic rise in LFTs correlated significantly (p = 0.009) with malignancy. Furthermore, premalignant pathology was found an additional 47% of the time. Seven patients ultimately chose an operation over continued observation for radiographic changes (mean 2.6 years), but none had cancer. In the last 2 years, we have evaluated 132 new patients with APLs, representing 47% of total referrals for pancreatic conditions. Nearly half were operated upon, with a 3:2 ratio of solid to cystic lesions. This differs significantly (p = 0.037) from the previous 3 years (2:3 ratio), reflecting tolerance for cysts <3 cm and side-branch IPMN. Surgery was undertaken more often when a solid APL was encountered (74%) than for cysts (32%). Some solid APLs were actually unresectable cancers. Due to anxiety, two patients requested an operation over continued observation, and neither had cancer. CONCLUSION: APLs occur commonly, are often solid, and reflect a spectrum of diagnoses. Sendai guidelines are not transferable to solid masses but have safely refined management of cysts. An asymptomatic rise in LFTs cannot be overlooked nor should a patient or doctor's anxiety, given the prevalence of cancer in APLs. PMID- 19130155 TI - The ileal pouch anal anastomosis: to divert or not to divert? The case for diversion. AB - The ileal pouch anal anastomosis is a high-risk anastomosis with a high rate of complications. Because of this proximal diversion has often been done when performing this operation. Recently the practice of proximal diversion has been questioned, noting the high rate of complications associated with the stoma and stomal reversal. However, the cumulative data does suggest that an undiverted ileal pouch anal anastomosis has a higher leak rate and is associated with increased pelvic sepsis. Because of the severity of this complication proximal diversion is still advocated in the majority of cases. PMID- 19130154 TI - Management of ERCP-related perforations: outcomes of single institution in Korea. AB - INTRODUCTION: The aim of this study was to analyze clinicoradiologic findings and treatment outcomes of patients with endoscopic retrograde cholangiopancreatography (ERCP)-related perforations. Between May 2003 and November 2007, 2,247 ERCP procedures with or without sphincterotomy were performed at Ajou University Medical Center, Suwon, Korea, and 20 perforations (0.89%) were identified. DISCUSSION: We retrospectively reviewed medical and surgical records of each patient. Of 18 patients, 11 patients (61.1%) underwent nonsurgical management, and seven patients (38.9%) received surgical management. There were no significant differences in age, gender, and laboratory findings between two groups (P > 0.05). The hospital stay was significantly longer in the operative group than that of the conservative group (P < 0.05, respectively). The most common cause of perforation was sphincterotomy (n = 8) in the conservative group whereas scope itself (n = 6) in operative group, showing a significant difference between the two groups (P < 0.05). The retroperitoneal air was most common findings in eight patients (72.7%) of the conservative group, while six (85.7%) patients of the operative group presented with intraperitoneal air, displaying a significant difference in location of air between the two groups (P < 0.05). Most of sphincterotomy-related perforations were managed nonsurgically. However, the scope-related perforations were usually large and required immediate surgery. Moreover, the delayed operation resulted in a longer hospital stay and high morbidity. Therefore, the selective early surgical intervention is suggested when scope-related perforations are discovered. PMID- 19130157 TI - The drop toe sign: an indicator of neurologic impairment in congenital clubfoot. AB - Nine patients presenting during infancy were identified with clubfeet and absent anterior and lateral compartment functions. We considered these to be neurogenic clubfeet. All patients had the drop toe sign: resting posture of the toes in plantarflexion and absent active dorsiflexion movement after plantar stimulation of the foot. Two patients (three feet) underwent exploration of the peroneal nerve, which revealed anatomic abnormalities. Six patients required more casts than typical for initial correction of deformity; all but two had Achilles tenotomy. Four relapsed despite full-time bracing and eventually needed intraarticular surgery to achieve a plantigrade foot. Idiopathic absent peroneal nerve function is not a well-described entity in the clubfoot literature. All babies with clubfoot should be examined for the drop toe sign. When noted, the feet will likely be more difficult to correct initially, may need early Achilles tendon lengthening, will likely need permanent bracing, are likely to relapse and need intraarticular surgery, and may need multiple surgeries to remain plantigrade throughout growth. LEVEL OF EVIDENCE: Level IV, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19130158 TI - Ponseti treatment for idiopathic clubfoot: minimum 5-year followup. AB - Ponseti clubfoot treatment has become more popular during the last decade. We reviewed the medical records of 74 consecutive infants (117 club feet) who underwent Ponseti treatment. Minimum followup was 5 years (mean, 6.3 years; range, 5-9 years). We studied age at presentation, previous treatment, the initial severity score of the Pirani scoring system, number of casts, need for Achilles tenotomy or other surgical procedures, and brace use. We measured final ankle motion and parents' perception of outcome. Late presentation and previous non-Ponseti treatment were associated with lower initial severity score, fewer casts, and less need for tenotomy. Forty-four percent of patients had poor brace use. We observed better brace use (75%) in babies who presented late for treatment. Good brace use predicted less need for extensive surgical procedures. Twenty-four (32%) babies underwent additional surgical procedures other than tenotomy, including 21% who underwent tibialis anterior tendon transfer. At followup, 89% of feet had adequate dorsiflexion (5 degrees or greater). Parents indicated high satisfaction with the treatment results. Ankle motion was not associated with parents' satisfaction. The Ponseti method is effective, even if treatment starts late or begins after failure at other centers. Brace use influenced the success of treatment. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19130159 TI - Acetabular morphology: implications for joint-preserving surgery. AB - Appropriate anatomic concepts for surgery to treat femoroacetabular impingement require a precise appreciation of the native acetabular anatomy. We therefore determined (1) the spatial acetabular rim profile, (2) the topography of the articular lunate surface, and (3) the 3-D relationships of the acetabular opening plane comparing 66 bony acetabula from 33 pelves in female and male pelves. The acetabular rim profile had a constant and regular wave-like outline without gender differences. Three prominences anterosuperiorly, anteroinferiorly and posteroinferiorly extended just above hemispheric level. Two depressions were below hemispheric level, of 9 degrees at the anterior wall and of 21 degrees along the posterosuperior wall. In 94% of all acetabula, the deepest extent of the articular surface was within 30 degrees of the anterosuperior acetabular sector. In 99% of men and in 91% of women, the depth of the articular surface was at least 55 degrees along almost half of the upper acetabular cup. The articular surface was smaller in women than in men. The acetabular opening plane was orientated in 21 degrees +/- 5 degrees for version, 48 degrees +/- 4 degrees for inclination and 19 degrees +/- 6 degrees for acetabular tilt with no gender differences. We defined tilt as forward rotation of the entire acetabular cup around its central axis; because of interindividual variability of acetabular tilt, descriptions of acetabular lesions during surgery, CT scanning and MRI should be defined and recorded in relation to the acetabular notch. Acetabular tilt and pelvic tilt should be separately identified. We believe this information important for surgeons performing rim trimming in FAI surgery or performing acetabular osteotomies. PMID- 19130160 TI - Clinical presentation of patients with symptomatic anterior hip impingement. AB - Femoroacetabular impingement (FAI) is considered a cause of labrochondral disease and secondary osteoarthritis. Nevertheless, the clinical syndrome associated with FAI is not fully characterized. We determined the clinical history, functional status, activity status, and physical examination findings that characterize FAI. We prospectively evaluated 51 patients (52 hips) with symptomatic FAI. Evaluation of the clinical history, physical exam, and previous treatments was performed. Patients completed demographic and validated hip questionnaires (Baecke et al., SF-12, Modified Harris hip, and UCLA activity score). The average patient age was 35 years and 57% were male. Symptom onset was commonly insidious (65%) and activity-related. Pain occurred predominantly in the groin (83%). The mean time from symptom onset to definitive diagnosis was 3.1 years. Patients were evaluated by an average 4.2 healthcare providers prior to diagnosis and inaccurate diagnoses were common. Thirteen percent had unsuccessful surgery at another anatomic site. On exam, 88% of the hips were painful with the anterior impingement test. Hip flexion and internal rotation in flexion were limited to an average 97 degrees and 9 degrees, respectively. The patients were relatively active, yet demonstrated restrictions of function and overall health. These data may facilitate diagnosis of this disorder. PMID- 19130161 TI - Limitations of structural allograft in revision total knee arthroplasty. AB - Management of large bone defects in total knee arthroplasty (TKA) usually has involved modular prostheses with metal augments, structural allografts, and megaprostheses. We retrospectively reviewed the outcome of treatment of major bone defects for 74 patients (79 knees) who had revision TKAs with structural allografts; nine patients were lost to followup before 5 years, leaving 65 patients (70 knees, or 88%) followed for a minimum of 5 years or until revision or death. Medical records, radiographs, patient surveys, and correspondence were used for all data. Sixteen patients (22.8%) had failed reconstructions and underwent additional revision surgery; eight of the 16 were secondary to allograft failure, three were secondary to failure of a component not supported by allograft, and five were secondary to infection. In patients not requiring revision surgery, the Knee Society score improved from 49 preoperatively to 87 postoperatively. We observed revision-free survival of 80.7% (95% confidence interval, 71.7-90.8) at 5 years and 75.9% (95% confidence interval, 65.6-87.8) at 10 years. Our data support the selective use of structural allograft for large cavitary defects encountered during TKA. However, the rates of complications and reoperations suggest efforts to improve results or develop more durable alternative methods are warranted for these challenging reconstructions. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19130162 TI - Cemented versus uncemented hemiarthroplasty for displaced femoral neck fractures. AB - Hemiarthroplasty is the most commonly used treatment for displaced femoral neck fractures in the elderly. There is limited evidence in the literature of improved functional outcome with cemented implants, although serious cement-related complications have been reported. We performed a randomized, controlled trial in patients 70 years and older comparing a cemented implant (112 hips) with an uncemented, hydroxyapatite-coated implant (108 hips), both with a bipolar head. The mean Harris hip score showed equivalence between the groups, with 70.9 in the cemented group and 72.1 in the uncemented group after 3 months (mean difference, 1.2) and 78.9 and 79.8 after 12 months (mean difference, 0.9). In the uncemented group, the mean duration of surgery was 12.4 minutes shorter and the mean intraoperative blood loss was 89 mL less. The Barthel Index and EQ-5D scores did not show any differences between the groups. The rates of complications and mortality were similar between groups. Both arthroplasties may be used with good results after displaced femoral neck fractures. PMID- 19130163 TI - Does regional anesthesia improve outcome after total knee arthroplasty? AB - Total knee arthroplasty (TKA) is amenable to various regional anesthesia techniques that may improve patient outcome. We sought to answer whether regional anesthesia decreased mortality, cardiovascular morbidity, deep venous thrombosis and pulmonary embolism, blood loss, duration of surgery, pain, opioid-related adverse effects, cognitive defects, and length of stay. We also questioned whether regional anesthesia improved rehabilitation. To do so, we performed a systematic review of the contemporary literature comparing general anesthesia and/or systemic analgesia with regional anesthesia and/or regional analgesia for TKA. To reflect contemporary surgical and anesthetic practice, only randomized, controlled trials from 1990 onward were included. We identified 28 studies involving 1538 patients. There was insufficient evidence from randomized, controlled trials alone to conclude if anesthetic technique influenced mortality, cardiovascular morbidity other than postoperative hypotension, or the incidence of deep venous thrombosis and pulmonary embolism when using thromboprophylaxis. Our review suggests there was no difference in perioperative blood loss or duration of surgery in patients who received general anesthesia versus regional anesthesia. Compared with general anesthesia and/or systemic analgesia, regional anesthesia and/or analgesia reduced postoperative pain, morphine consumption, and opioid-related adverse effects. Length of stay may be reduced and rehabilitation facilitated for patients undergoing regional anesthesia and analgesia for TKA. PMID- 19130164 TI - Animal models to study thyroid hormone action in cerebellum. AB - Thyroid hormone plays a crucial role in the development and functional maintenance of the central nervous system including the cerebellum. To study the molecular mechanisms of thyroid hormone action, various animal models have been used. These are classified: (1) congenital hypothyroid animals due to thyroid gland dysgenesis or thyroid dyshormonogenesis, (2) thyroid hormone receptor (TR) gene-mutated animals, and (3) thyroid hormone transport or metabolism-modified animals. TR is a ligand-activated transcription factor. In the presence of ligand, it activates transcription of target gene, whereas it represses the transcription without ligand. Thus, phenotype of TR-knockout mouse is different from that of hypothyroid animal (low thyroid hormone level), in which unliganded TR actively represses the transcription. On the other hand, human patient harboring mutant TR expresses different phenotypes depending on the function of mutated TR. To mimic this phenotype, other animal models are generated. In addition, recent human studies have shown that thyroid hormone transporters such as monocarboxylate transporter (MCT) 8 may play an important role in thyroid hormone-mediated brain development. However, MCT8 knockout mouse show different phenotypes from a human patient. This article introduces representative animal models currently used to study various aspects of thyroid hormone, particularly to study the involvement of the thyroid hormone system on the development and functional maintenance of the cerebellum. PMID- 19130165 TI - Myocardial perfusion SPECT in the diagnosis of apical hypertrophic cardiomyopathy. AB - BACKGROUND: Apical hypertrophy cardiomyopathy (ACM) is a rare condition characterized by asymmetric myocardial hypertrophy of the apex of the left ventricle. When two-dimensional echocardiography is limited by a poor acoustic window, patients are often referred for MRI. Our hypothesis is that a cheaper and more widely available diagnostic modality like myocardial perfusion single photon emission computed tomography (SPECT) may be helpful in the diagnosis of ACM. OBJECTIVE: The purpose of this prospective study was to define the characteristics of rest and stress SPECT studies in patients with known ACM, and whether SPECT may be helpful in the diagnosis of ACM. METHODS: Adult patients with ACM were enrolled in the study. Diagnosis was made with 2-D echo. A rest and exercise or dipyridamole stress SPECT study was performed in all patients with Tc 99m sestamibi. RESULTS: We enrolled 20 patients (mean age 60 +/- 16 years), 9 were female, with ACM. SPECT at rest revealed in 15 patients (75%) an increased apical tracer uptake, a spade-like deformity of the left ventricular chamber, and the "Solar Polar" map pattern consistent with ACM. The sensitivity, specificity, positive predictive value, and negative predictive value of SPECT for detecting ACM were 75%, 100%, 100%, and 80%, respectively. CONCLUSION: Three-fourths of adult patients with ACM showed, on myocardial perfusion SPECT, characteristic findings which were not seen in age-matched control subjects, such as a significant increased apical tracer uptake, a spade-like deformity of the left ventricle, and the "Solar Polar" map. Nuclear physicians should be aware of these SPECT findings because many ACM patients may first end up in the nuclear labs due to their markedly abnormal ECG for exclusion of obstructive coronary artery disease. PMID- 19130166 TI - Genetic analysis and pathogenicity of betanodavirus isolated from wild redspotted grouper Epinephelus akaara with clinical signs. AB - Diseased wild redspotted grouper Epinephelus akaara were collected from Seto Inland Sea, Ehime Prefecture, in August 2002. Fish showed erratic swimming behavior and inflation of the swim bladder. The fish brains were positive for nodavirus in both RT-PCR and nested PCR. The sequence of the nested PCR product (177 nt) was closely related to that of a known betanodavirus, redspotted grouper nervous necrosis virus. When juvenile sevenband grouper E. septemfasciatus were challenged intravitreously with virus, abnormal swimming behavior and high mortality were observed. This is the first report on viral nervous necrosis in a wild population of redspotted grouper with clinical signs. PMID- 19130167 TI - Restoration of potato virus X coat protein capacity for assembly with RNA after His-tag removal. AB - We found that the fusion of hexahistidine (H)6 tag to the potato virus X (PVX) coat protein (CP) abolished its ability to be assembled with viral RNA into helical nucleoprotein virus-like particles (VLPs). Instead, irregular agglomerates were produced upon incubation of PVX RNA with (H)6-tagged PVX CP. A factor Xa recognition site, IEGR, was inserted upstream of the CP coding sequence. Removal of the (H)6 tag from PVX CP by Xa protease restored its ability to bind RNA and to assemble VLPs. In addition to the canonical IEGR site, the factor Xa protease was found to cleave PVX CP at a second (non-consensus) site, AVTRGR, located close to the C-terminus of PVX CP. The latter cleavage did not affect reassembly of the PVX RNA and CP into VLPs. PMID- 19130168 TI - Intracellular localization of influenza C virus NS2 protein (NEP) in infected cells and its incorporation into virions. AB - RNA segment 7 of influenza C virus encodes two non-structural (NS) proteins, NS1 and NS2. The influenza C virus NS2 protein has been proposed to possess nuclear export activity like that of influenza A and B virus NS2 proteins (NEP). In the present study, we investigated the kinetics and localization of the NS2 protein in influenza C virus-infected cells, and analysed whether NS2 is present in virions. Immunofluorescent staining analysis of the infected cells indicated that NS2 was localized in the nucleus immediately after synthesis and predominantly in the cytoplasm in the later stages of infection. Confocal microscopy revealed that a part of the NS2 protein was colocalized with nucleoprotein NP/vRNP in the cytoplasm and on the cell membrane in the late stages of infection. The NS2 protein was detected in influenza C virions purified by gradient centrifugations and/or affinity chromatography. Trypsin treatment demonstrated that the NS2 protein was present inside the viral envelope. Furthermore, glycerol gradient analysis of detergent-solubilized virions revealed that the NS2 protein cosedimented with vRNPs. These data suggest that the influenza C virus NS2 protein is incorporated into virions, where it associates with vRNP. PMID- 19130169 TI - Pathological lesions and viral localization of influenza A (H5N1) virus in experimentally infected Chinese rhesus macaques: implications for pathogenesis and viral transmission. AB - Chinese rhesus macaques infected with influenza virus A/Tiger/Harbin/01/2002 (H5N1) developed acute interstitial pneumonia with diffuse alveolar damage. The results of virus isolation, reverse transcriptase polymerase chain reaction, immunohistochemistry, and in situ hybridization showed that the lung was the major target organ of the H5N1 virus infection. No virus was detected in the extrapulmonary organs. The results of immunohistochemistry and in situ hybridization also showed that pneumocytes and macrophages of the lower airway, not the ciliary epithelium of the trachea and bronchi, were the chief target cells in the lung tissue of the infected Chinese rhesus macaque. Our data indicate that the Chinese rhesus macaque is suitable as a new primate model for H5N1 virus research, especially for the study of H5N1 virus transmission. The predilection of the H5N1 virus to infect the lower airway suggests that the failure of the virus to attach to the ciliary epithelium of the trachea and bronchi may be a limiting factor in human-to-human transmissibility of the H5N1 virus. PMID- 19130170 TI - Glutamine, arginine, and leucine signaling in the intestine. AB - Glutamine and leucine are abundant constituents of plant and animal proteins, whereas the content of arginine in foods and physiological fluids varies greatly. Besides their role in protein synthesis, these three amino acids individually activate signaling pathway to promote protein synthesis and possibly inhibit autophagy-mediated protein degradation in intestinal epithelial cells. In addition, glutamine and arginine stimulate the mitogen-activated protein kinase and mammalian target of rapamycin (mTOR)/p70 (s6) kinase pathways, respectively, to enhance mucosal cell migration and restitution. Moreover, through the nitric oxide-dependent cGMP signaling cascade, arginine regulates multiple physiological events in the intestine that are beneficial for cell homeostasis and survival. Available evidence from both in vitro and in vivo animal studies shows that glutamine and arginine promote cell proliferation and exert differential cytoprotective effects in response to nutrient deprivation, oxidative injury, stress, and immunological challenge. Additionally, when nitric oxide is available, leucine increases the migration of intestinal cells. Therefore, through cellular signaling mechanisms, arginine, glutamine, and leucine play crucial roles in intestinal growth, integrity, and function. PMID- 19130171 TI - Diffuse large B-cell lymphoma with florid hemophagocytosis. PMID- 19130172 TI - Correlation between characteristics of unrelated bone marrow donor and cell density of total nucleated cell in bone marrow harvest. AB - The relationship between the features of bone marrow donor and the quality of marrow harvest has been unclear because most of bone marrow registries have multiple collection centers with somewhat different harvest procedures. We are able to address this issue for Tzu Chi General Hospital is the only collection center affiliated with Tzu Chi Taiwan Bone Marrow Registry. Between November 1997 and March 2002, data of 286 healthy unrelated donors was analyzed to correlate with the cell density of total nucleated cell in bone marrow harvests. The harvest procedure was standardized by single-hole harvest needle under general anesthesia. The operation staffs were restricted within the members of Oncology Hematology division. The results showed that the cell density of bone marrow harvest was positively correlated with donor body weight and peripheral white blood cell count P = 0.0475, P < 0.0001, but negatively correlated with the total volume of bone marrow harvest P < 0.0001. We recommend that if multiple human leukocyte antigen-matched donors are available, donor with higher body weight and/or higher white blood cell count be selected for allogeneic bone marrow transplantation. PMID- 19130173 TI - Successful treatment with liposomal doxorubicin for widespread Kaposi's sarcoma and human herpesvirus-8 related severe hemophagocytic syndrome in a patient with acquired immunodeficiency syndrome. AB - Hemophagocytic syndrome (HPS) sometimes occurres in patients with acquired immunodeficiency syndrome (AIDS). Human herpesvirus-8 (HHV-8)/Kaposi's sarcoma (KS)-associated herpesvirus has so far been recognized as a trigger of HPS in immunosuppressed subject. We describe a 39-year-old man with AIDS who had widespread mucocutaneous and pulmonary KS and severe HPS. No opportunistic infections or neoplasias were detected except for KS. HHV-8-DNA could be detected in this patient by polymerase chain reaction (PCR) in the serum. Clinical symptoms and cytopenia originating from HPS were reduced by pulse therapy of corticosteroid, antibiotics, and virucides, but recurred with dose reduction of the steroid. Mucocutaneous tumors, edema, and dyspnea had progressed rapidly at this time. Liposomal doxorubicin was given and showed marked effects on both mucocutaneous and plural tumors. HPS also subsided and the serum HHV-8 DNA level markedly decreased after initial treatment with liposomal doxorubicin. HHV-8 clearance with liposomal doxorubicin has recently been reported. Liposomal doxorubicin suppressed not only the widespread KS tumors, but also HHV-8 viremia resulting in decreased HPS in this patient. PMID- 19130174 TI - 25 years of the antiphospholipid syndrome: lights and shadows on the primary prophylaxis of asymptomatic patients with confirmed positivity of antiphospholipid antibodies. PMID- 19130176 TI - Contemporary issues on clopidogrel therapy. AB - In this paper, data from available studies regarding some contemporary issues on clopidogrel therapy are analyzed. In particular, the following clinical questions have been considered and addressed: (a) Is early clopidogrel treatment needed in patients with acute coronary syndromes treated medically or undergoing percutaneous coronary intervention (PCI)? (b) What is the optimal clopidogrel loading dose in patients undergoing PCI? (c) Is pre-treatment with clopidogrel before PCI needed, or can clopidogrel loading be given in the catheter laboratory before intervention, but after coronary anatomy is known? (d) What is the optimal clopidogrel strategy in patients on chronic clopidogrel therapy undergoing PCI? (e) Does the degree of clopidogrel response influence clinical outcome in patients undergoing PCI? PMID- 19130177 TI - Anticoagulation for portal vein thrombosis in cirrhotic patients should be always considered. PMID- 19130178 TI - Serum IL-6 levels are associated with significant coronary stenosis in cardiovascularly asymptomatic inner-city black adults in the US. AB - OBJECTIVES AND DESIGN: The objective of this study was to explore whether increased levels of inflammatory cytokines are associated with the risk of clinically silent coronary artery disease. SUBJECTS: Three-hundred-fifty-six black adults aged 25-54 residing in inner city of Baltimore, Maryland, United States were included in this study. METHODS: Sociodemographics were assessed as were lipid profiles, IL-6, tumor necrosis factor-alpha (TNF-alpha), soluble intercellular adhesion molecule-1 (sICAM-1), and high-sensitivity C-reactive protein (hs-CRP) levels. Computed tomography (CT) coronary angiography was performed. RESULTS: Coronary calcification was identified in 22.5 % participants and 14 % had significant (>or=50 %) coronary stenosis. Multiple logistic regression analyses suggested that IL-6 levels were independently associated with the presence of coronary calcification and significant coronary stenosis, while TNF-alpha, sICAM-1 and hs-CRP levels were not. CONCLUSIONS: This study underscores a critical role for IL-6 in atherosclerosis and suggests that IL-6 may be a marker for significant coronary stenosis in cardiovascularly asymptomatic individuals. PMID- 19130179 TI - Characterization of the yellow fever mosquito sterol carrier protein-2 like 3 gene and ligand-bound protein structure. AB - The sterol carrier protein-2 like 3 gene (AeSCP-2L3), a new member of the SCP-2 protein family, is identified from the yellow fever mosquito, Aedes aegypti. The predicted molecular weight of AeSCP-2L3 is 13.4 kDa with a calculated pI of 4.98. AeSCP-2L3 transcription occurs in the larval feeding stages and the mRNA levels decrease in pupae and adults. The highest levels of AeSCP-2L3 gene expression are found in the body wall, and possibly originated in the fat body. This is the first report of a mosquito SCP-2-like protein with prominent expression in tissue other than the midgut. The X-ray protein crystal structure of AeSCP-2L3 reveals a bound C16 fatty acid whose acyl tail penetrates deeply into a hydrophobic cavity. Interestingly, the ligand-binding cavity is slightly larger than previously described for AeSCP-2 (Dyer et al. J Biol Chem 278:39085-39091, 2003) and AeSCP 2L2 (Dyer et al. J Lipid Res M700460-JLR200, 2007). There are also an additional 10 amino acids in SCP-2L3 that are not present in other characterized mosquito SCP-2s forming an extended loop between beta 3 and beta 4. Otherwise, the protein backbone is exceedingly similar to other SCP-2 and SCP-2-like proteins. In contrast to this observed high structural homology of members in the mosquito SCP2 family, the amino acid sequence identity between the members is less than 30%. The results from structural analysis imply that there have been evolutionary constraints that favor the SCP-2 C(alpha) backbone fold while the specificity of ligand binding can be altered. PMID- 19130180 TI - Hypertriglyceridemia: phenomics and genomics. AB - Hypertriglyceridemia is a common complex metabolic trait that is associated with increased atherosclerosis risk, presence of the metabolic syndrome and, with extreme elevation, increased risk of pancreatitis. Hierarchical cluster analysis using clinical and biochemical features of the Frederickson hyperlipoproteinemia types can generate hypotheses for molecular genetic studies. High throughput resequencing of individuals at the extremes of plasma triglyceride concentration has shown that both rare genetic variants with large effects and common genetic variants with moderate effects explain a relatively large proportion of variation. Very recent progress using high-density sets of genome-wide markers have identified additional genetic determinants of plasma triglyceride concentrations, albeit within largely normolipidemic subjects and with small effect sizes. Phenomic evaluation of patients with hypertriglyceridemia might help to clarify genotype-phenotype correlations and responses to interventions. PMID- 19130181 TI - Improvement of cerebral function by anti-amyloid precursor protein antibody infusion after traumatic brain injury in rats. AB - We previously demonstrated the increased amyloid precursor protein (APP) immunoreactivity around the site of damage after traumatic brain injury (TBI). However, the function of APP after TBI has not been evaluated. In this study, we investigated the effects of direct infusion of an anti-APP antibody into the damaged brain region on cerebral function and morphological changes following TBI in rats. Three days after TBI, there were many TUNEL-positive neurons and astrocytes around the damaged region and a significantly greater number of TUNEL positive cells in the PBS group compared with the anti-APP group found. Seven days after TBI, there were significantly a greater number of large glial fibrillary acidic protein-positive cells, long elongated projections, and microtubule-associated protein-2-positive cells around the damaged region in the anti-APP group compared with the PBS group found. Seven days after TBI, the region of brain damage was significantly smaller and the time to arrival at a platform was significantly shorter in the anti-APP group compared with the PBS group. Furthermore, after TBI in the anti-APP group, the time to arrival at the platform recovered to that observed in uninjured sham operation group rats. These data suggest that the overproduction of APP after TBI inhibits astrocyte activity and reduces neural cell survival around the damaged brain region, which speculatively may be related to the induction of Alzheimer disease-type dementia after TBI. PMID- 19130183 TI - Lipid-activated transcription factors control bile acid glucuronidation. AB - Bile acids subserve important physiological functions in the control of cholesterol homeostasis. Indeed, hepatic bile acid synthesis and biliary excretion constitute the main route for cholesterol removal from the human body. On the other hand, bile acids serve as natural detergents for the intestinal absorption of dietary cholesterol. However, due to their detergent properties, bile acids are inherently cytotoxic, and their cellular level may be tightly controlled to avoid pathological situations such as cholestasis. Recent investigations have illustrated the crucial roles that a series of ligand activated transcription factors has in the control of hepatic bile acids synthesis, transport and metabolism. Thus, the lipid-activated nuclear receptors, farnesoid X-receptor (FXR), liver X-receptor (LXR), pregnane X-receptor (PXR) and peroxisome proliferator-activated receptor alpha (PPAR alpha), modulate the expression and activity of genes controlling bile acid homeostasis in the liver. Several members of the UDP-glucuronosyltransferase (UGT) enzymes family are among the bile acid metabolizing enzymes regulated by these receptors. UGTs catalyze glucuronidation, a major phase II metabolic reaction, which converts hydrophobic bile acids into polar and urinary excretable metabolites. This article summarizes our recent observations on the regulation of bile acid conjugating UGTs upon pharmacological activation of lipid-activated receptors, with a particular interest for the role of PPAR alpha and LXRalpha in controlling human UGT1A3 expression. PMID- 19130185 TI - Implications of global and local mobility in amorphous sucrose and trehalose as determined by differential scanning calorimetry. AB - PURPOSE: To investigate the local and global mobility in amorphous sucrose and trehalose and their potential implications on physical stability. METHODS: Amorphous sucrose was prepared by lyophilization while amorphous trehalose was prepared by dehydration of trehalose dihydrate. The variation in the effective activation energy of alpha-relaxation through glass transition has been determined by applying an isoconversional method. Beta-relaxations were detected as shallow peaks, at temperatures below the glass transition temperature, caused by annealing glassy samples at different temperatures and subsequently heating at different rates in a differential scanning calorimeter. The effect of heating rate on the beta-relaxation peak temperature formed the basis for the calculation of the activation energy. RESULTS: alpha-Relaxations in glassy trehalose were characterized by larger activation energy barrier compared to sucrose, attributable to a more compact molecular structure of trehalose. The effect of temperature on viscous flow was greater in trehalose which can have implications on lyophile collapse. The size of the cooperatively rearranging regions was about the same for sucrose and trehalose suggesting similar dynamic heterogeneity at their respective glass transition temperatures. The activation energy of beta relaxations increased with annealing temperature due to increasing cooperative motions and the increase was larger in sucrose. The temperature at which beta relaxation was detected for a given annealing time was much less in sucrose implying that progression of local motions to cooperative motions occurred at lower temperatures in sucrose. CONCLUSIONS: Trehalose, having a lower free volume in the glassy state due to a more tightly packed molecular structure, is characterized by larger activation energies of alpha-relaxation and experiences a greater effect of temperature on the reduction in the activation energy barrier for viscous flow. The pronounced increase in cooperative motions in sucrose upon annealing at temperatures below (T(g) -50) suggest that even a small excursion in temperature could result in a significant increase in mobility. PMID- 19130182 TI - Circulatory lipid transport: lipoprotein assembly and function from an evolutionary perspective. AB - Circulatory transport of neutral lipids (fat) in animals relies on members of the large lipid transfer protein (LLTP) superfamily, including mammalian apolipoprotein B (apoB) and insect apolipophorin II/I (apoLp-II/I). Latter proteins, which constitute the structural basis for the assembly of various lipoproteins, acquire lipids through microsomal triglyceride transfer protein (MTP)--another LLTP family member--and bind them by means of amphipathic structures. Comparative research reveals that LLTPs have evolved from the earliest animals and additionally highlights the structural and functional adaptations in these lipid carriers. For instance, in contrast to mammalian apoB, the insect apoB homologue, apoLp-II/I, is post-translationally cleaved by a furin, resulting in their appearance of two non-exchangeable apolipoproteins in the insect low-density lipoprotein (LDL) homologue, high-density lipophorin (HDLp). An important difference between mammalian and insect lipoproteins relates to the mechanism of lipid delivery. Whereas in mammals, endocytic uptake of lipoprotein particles, mediated via members of the LDL receptor (LDLR) family, results in their degradation in lysosomes, the insect HDLp was shown to act as a reusable lipid shuttle which is capable of reloading lipid. Although the recent identification of a lipophorin receptor (LpR), a homologue of LDLR, reveals that endocytic uptake of HDLp may constitute an additional mechanism of lipid delivery, the endocytosed lipoprotein appears to be recycled in a transferrin like manner. Binding studies indicate that the HDLp-LpR complex, in contrast to the LDL-LDLR complex, is resistant to dissociation at endosomal pH as well as by treatment with EDTA mimicking the drop in Ca(2+) concentration in the endosome. This remarkable stability of the ligand-receptor complex may provide a crucial key to the recycling mechanism. Based on the binding and dissociation capacities of mutant and hybrid receptors, the specific binding interaction of the ligand binding domain of the receptor with HDLp was characterized. These structural similarities and functional adaptations of the lipid transport systems operative in mammals and insects are discussed from an evolutionary perspective. PMID- 19130186 TI - Comparing models for perfluorooctanoic acid pharmacokinetics using Bayesian analysis. AB - Selecting the appropriate pharmacokinetic (PK) model given the available data is investigated for perfluorooctanoic acid (PFOA), which has been widely analyzed with an empirical, one-compartment model. This research examined the results of experiments [Kemper R. A., DuPont Haskell Laboratories, USEPA Administrative Record AR-226.1499 (2003)] that administered single oral or iv doses of PFOA to adult male and female rats. PFOA concentration was observed over time; in plasma for some animals and in fecal and urinary excretion for others. There were four rats per dose group, for a total of 36 males and 36 females. Assuming that the PK parameters for each individual within a gender were drawn from the same, biologically varying population, plasma and excretion data were jointly analyzed using a hierarchical framework to separate uncertainty due to measurement error from actual biological variability. Bayesian analysis using Markov Chain Monte Carlo (MCMC) provides tools to perform such an analysis as well as quantitative diagnostics to evaluate and discriminate between models. Starting from a one compartment PK model with separate clearances to urine and feces, the model was incrementally expanded using Bayesian measures to assess if the expansion was supported by the data. PFOA excretion is sexually dimorphic in rats; male rats have bi-phasic elimination that is roughly 40 times slower than that of the females, which appear to have a single elimination phase. The male and female data were analyzed separately, keeping only the parameters describing the measurement process in common. For male rats, including excretion data initially decreased certainty in the one-compartment parameter estimates compared to an analysis using plasma data only. Allowing a third, unspecified clearance improved agreement and increased certainty when all the data was used, however a significant amount of eliminated PFOA was estimated to be missing from the excretion data. Adding an additional PK compartment reduced the unaccounted-for elimination to amounts comparable to the cage wash. For both sexes, an MCMC estimate of the appropriateness of a model for a given data type, the Deviance Information Criterion, indicated that this two-compartment model was better suited to describing PFOA PK. The median estimate was 142.1 +/- 37.6 ml/kg for the volume of the primary compartment and 1.24 +/- 1.1 ml/kg/h for the clearances of male rats and 166.4 +/- 46.8 ml/kg and 30.3 +/- 13.2 ml/kg/h, respectively for female rats. The estimates for the second compartment differed greatly with gender-volume 311.8 +/- 453.9 ml/kg with clearance 3.2 +/- 6.2 for males and 1400 +/- 2507.5 ml/kg and 4.3 +/- 2.2 ml/kg/h for females. The median estimated clearance was 12 +/- 6% to feces and 85 +/- 7% to urine for male rats and 8 +/- 6% and 77 +/- 9% for female rats. We conclude that the available data may support more models for PFOA PK beyond two-compartments and that the methods employed here will be generally useful for more complicated, including PBPK, models. PMID- 19130187 TI - Drug-drug interaction predictions with PBPK models and optimal multiresponse sampling time designs: application to midazolam and a phase I compound. Part 2: clinical trial results. AB - PURPOSE: To compare results of population PK analyses obtained with a full empirical design (FD) and an optimal sparse design (MD) in a Drug-Drug Interaction (DDI) study aiming to evaluate the potential CYP3A4 inhibitory effect of a drug in development, SX, on a reference substrate, midazolam (MDZ). Secondary aim was to evaluate the interaction of SX on MDZ in the in vivo study. Methods To compare designs, real data were analysed by population PK modelling technique using either FD or MD with NONMEM FOCEI for SX and with NONMEM FOCEI and MONOLIX SAEM for MDZ. When applicable a Wald test was performed to compare model parameter estimates, such as apparent clearance (CL/F), across designs. To conclude on the potential interaction of SX on MDZ PK, a Student paired test was applied to compare the individual PK parameters (i.e. log(AUC) and log(C(max))) obtained either by a non-compartmental approach (NCA) using FD or from empirical Bayes estimates (EBE) obtained after fitting the model separately on each treatment group using either FD or MD. RESULTS: For SX, whatever the design, CL/F was well estimated and no statistical differences were found between CL/F estimated values obtained with FD (CL/F = 8.2 l/h) and MD (CL/F = 8.2 l/h). For MDZ, only MONOLIX was able to estimate CL/F and to provide its standard error of estimation with MD. With MONOLIX, whatever the design and the administration setting, MDZ CL/F was well estimated and there were no statistical differences between CL/F estimated values obtained with FD (72 l/h and 40 l/h for MDZ alone and for MDZ with SX, respectively) and MD (77 l/h and 45 l/h for MDZ alone and for MDZ with SX, respectively). Whatever the approach, NCA or population PK modelling, and for the latter approach, whatever the design, MD or FD, comparison tests showed that there was a statistical difference (P < 0.0001) between individual MDZ log(AUC) obtained after MDZ administration alone and co administered with SX. Regarding C(max), there was a statistical difference (P < 0.05) between individual MDZ log(C(max)) obtained under the 2 administration settings in all cases, except with the sparse design with MONOLIX. However, the effect on C(max) was small. Finally, SX was shown to be a moderate CYP3A4 inhibitor, which at therapeutic doses increased MDZ exposure by a factor of 2 in average and almost did not affect the C(max). CONCLUSION: The optimal sparse design enabled the estimation of CL/F of a CYP3A4 substrate and inhibitor when co administered together and to show the interaction leading to the same conclusion as the full empirical design. PMID- 19130189 TI - Fabrication and spectroscopic characterization of Langmuir-Blodgett films with luminescent rare earth complexes of long chain double functional ligands mono-L phthalate (L = hexadecyl, octadecyl and eicosyl). AB - In this paper, some novel long chain amphiphillic monoester molecules were designed to afford double functions: film-formation ability and luminescent sensitization ability. Subsequently organized molecular films of rare earth complexes with these functional ligands formulated as ML2NO3 were fabricated by the Langmuir-Blodgett film (LB) technology, where RE denotes rare earth ions Eu3+, Tb3+ and Dy3+; L denotes the long chain carboxylic ligands monohexadecyl phthalate (16-Phth), monooctadecyl phthalate (18-Phth) and monoeicosyl phthalate (20-Phth). The average molecular area was obtained according to the pi-A isotherms. The layer structure of the LB films was demonstrated by low-angle X ray diffraction and the average layer spacing was determined from the Bragg equation. UV absorption intensity increases linearly with the number of LB films layers, which indicates that the LB films are homogeneously deposited. The fluorescence spectra of these LB films were quite different from those of their solid complexes. It reveals that the long chain ester ligands are suitable for the excited states of Tb3+ and Dy3+ in the LB films as well as in the solid complexes, but not match with the europium ion in the LB films. PMID- 19130188 TI - Drug-drug interaction predictions with PBPK models and optimal multiresponse sampling time designs: application to midazolam and a phase I compound. Part 1: comparison of uniresponse and multiresponse designs using PopDes. AB - PURPOSE: To determine the optimal sampling time design of a drug-drug interaction (DDI) study for the estimation of apparent clearances (CL/F) of two co administered drugs (SX, a phase I compound, potentially a CYP3A4 inhibitor, and MDZ, a reference CYP3A4 substrate) without any in vivo data using physiologically based pharmacokinetic (PBPK) predictions, population PK modelling and multiresponse optimal design. METHODS: PBPK models were developed with AcslXtreme using only in vitro data to simulate PK profiles of both drugs when they were co administered. Then, using simulated data, population PK models were developed with NONMEM and optimal sampling times were determined by optimizing the determinant of the population Fisher information matrix with PopDes using either two uniresponse designs (UD) or a multiresponse design (MD) with joint sampling times for both drugs. Finally, the D-optimal sampling time designs were evaluated by simulation and re-estimation with NONMEM by computing the relative root mean squared error (RMSE) and empirical relative standard errors (RSE) of CL/F. RESULTS: There were four and five optimal sampling times (=nine different sampling times) in the UDs for SX and MDZ, respectively, whereas there were only five sampling times in the MD. Whatever design and compound, CL/F was well estimated (RSE < 20% for MDZ and <25% for SX) and expected RSEs from PopDes were in the same range as empirical RSEs. Moreover, there was no bias in CL/F estimation. Since MD required only five sampling times compared to the two UDs, D optimal sampling times of the MD were included into a full empirical design for the proposed clinical trial. A joint paper compares the designs with real data. CONCLUSION: This global approach including PBPK simulations, population PK modelling and multiresponse optimal design allowed, without any in vivo data, the design of a clinical trial, using sparse sampling, capable of estimating CL/F of the CYP3A4 substrate and potential inhibitor when co-administered together. PMID- 19130190 TI - Trends in lung cancer death rates in Belgium and The Netherlands: a systematic analysis of temporal patterns. AB - Belgium and The Netherlands have fairly similar smoking prevalence patterns, but distinct tobacco control policies. It is our aim to use lung cancer death rates, especially among the youngest age groups (30-39 years), as indicators of past smoking behavioral patterns to evaluate recent tobacco control efforts in both countries. Lung cancer mortality rates from 1954 to 1997 and from 1950 to 2000 were investigated in Belgium and The Netherlands, respectively, using the joinpoint regression modeling technique (log-linear Poisson models) to calculate annual percent change in death rate. In the most recent period (1984-2000) overall male lung cancer death rates have been declining at a faster rate in The Netherlands than in Belgium. In contrast, overall female lung cancer death rates (between 1950 and 2000) have been increasing at a faster rate in The Netherlands than in Belgium. Since 1988, however, APCs in death rates among Dutch females have begun to level off. Interestingly, during this same period, a significant annual decline of 7.7% among the youngest Dutch women (30-39 years) has been observed. Tobacco use prevention and interventions seem to have an impact on smoking prevalence, especially among younger age groups. In The Netherlands, where aggressive anti-tobacco campaigns were introduced a few years earlier than in Belgium, male lung cancer mortality rates have been declining more rapidly, and female lung cancer mortality rates have begun to level off. PMID- 19130191 TI - Methodological complexities and the use of GIS in conducting a community needs assessment of a large U.S. municipality. AB - Reliable assessment of health and social needs within a community ensures that local input drives strategic planning and programmatic decisions. Results are used to determine the priorities and focus of local public health departments and to support the activities of health and social service agencies. We utilized a geographic information system in the planning, administration, and analysis of a recent community needs assessment to ensure a reliable and randomly distributed sample of a diverse urban population and to allow for small geographic area analysis of disease prevalence data. Collection of location data at the time of the 3,361 face-to-face assessment interviews allowed for the association of spatial data with measures of health conditions and behaviors. Results of the assessment indicated a need for public health programs designed to address emerging public health concerns in the community. Incorporating a spatial component in analyses of morbidity and needs assessment data allows for efficient integration of demographic and socio-economic census data and permits ad hoc analyses of varied and changing geographic strata. PMID- 19130192 TI - Prediction of catalytic residues using the variation of stereochemical properties. AB - In this paper, we investigate a simple protein sequence conservation measure which takes amino acid similarity into account. Instead of grouping 20 amino acids into disjoint sets in previous methods, we consider ten overlapping classes. The method is based on the assumption that a column in a multiple sequence alignment is evolved from an identical column in the evolutionary history. Two ten-dimensional vectors are constructed for each position to denote frequencies of ten classes in a column and the corresponding hypothetical identical column. Then the cosine function of the angle between these two vectors is considered as a measure of divergence of stereochemical properties at this position. This divergence, combining with other conservation scores, is used as conservation measure of the column. Finally, we evaluate our methods by identifying catalytic sites, using rank analysis criterion and receiver operator characteristic analysis criterion. PMID- 19130193 TI - Single nucleotide polymorphism that accompanies a missense mutation (Gln488His) impedes the dimerization of Hsp90. AB - A single nucleotide polymorphism (SNP) that causes a missense mutation of highly conserved Gln488 to His of the alpha isoform of the 90-kDa heat shock protein (Hsp90alpha) molecular chaperone is observed in Caucasians. The mutated Hsp90alpha severely reduced the growth of yeast cells. To investigate this molecular mechanism, we examined the domain-domain interactions of human Hsp90alpha by using bacterial 2-hybrid system. Hsp90alpha was expressed as a full length form, N-terminal domain (residues 1-400), or middle (residues 401-617) plus C-terminal (residues 618-732) domains (MC domain/amino acids 401-732). The Gln488His substitution in MC domain did not affect the intra-molecular interaction with N-terminal domain, whereas the dimeric interaction-mediated by the inter-molecular interaction between MC domains was decreased to 32%. Gln488Ala caused a similar change, whereas Gln488Thr, which exceptionally occurs in mitochondrial Hsp90 paralog, fully maintained the dimeric interaction. Therefore, the SNP causing Gln488His mutation could abrogate the Hsp90 function due to reduced dimerization. PMID- 19130194 TI - Molecular dynamics study of the structure, flexibility and dynamics of thermostable l1 lipase at high temperatures. AB - Molecular Dynamics (MD) simulations have been used to understand how protein structure, dynamics, and flexibility are affected by adaptation to high temperature for several years. We report here the results of the high temperature MD simulations of Bacillus stearothermophilus L1 (L1 lipase). We found that the N terminal moiety of the enzyme showed a high flexibility and dynamics during high temperature simulations which preceded and followed by clear structural changes in two specific regions; the small domain and the main catalytic domain or core domain of the enzyme. These two domains interact with each other through a Zn(2+) binding coordination with Asp-61 and Asp-238 from the core domain and His-81 and His-87 from the small domain. Interestingly, the His-81 and His-87 were among the highly fluctuated and mobile residues at high temperatures. The results appear to suggest that tight interactions of Zn(2+)-binding coordination with specified residues became weak at high temperature which suggests the contribution of this region to the thermostability of the enzyme. PMID- 19130195 TI - An anionic porphyrin binds beta-lactoglobulin A at a superficial site rich in lysine residues. AB - Binding of small ligands to globular proteins remains a major research topic in biophysics. We have studied the binding of several photoactive dyes to beta lactoglobulin (BLG), as a model to investigate the photoinduced effects of porphyrins on proteins. A combination of optical spectroscopies (fluorescence, circular dichroism) and molecular docking simulations were used to estimate the pH-dependence of the binding parameters and the docking location for meso tetrakis(sulfonatophenyl)-porphyrin (TPPS). We have observed that the binding of TPPS is not modulated by the pH-mediated conformational transition of the protein (i.e., Tanford transition). Binding of TPPS appears to occur with some degree of negative cooperativity. Moreover, TPPS remains bound even upon partial denaturation of the protein. These results are consistent with a superficial binding site at a location removed from the aperture of the interior beta-barrel. Binding occurs through electrostatic interactions between the negative SO(3) (-) groups of TPPS and positively charged Lys and Arg residues. This is the first study that explores the interaction of an anionic porphyrin with BLGA in a pH range that spans across the Tanford transition. Establishing the location of the binding site will enable us to explain the photoinduced conformational effects mediated by TPPS on BLG. PMID- 19130196 TI - Hepatitis C virus infection and the brain. AB - There is growing evidence that hepatitis C virus (HCV)-infection may affect the brain. About half of the HCV-infected patients complain of chronic fatigue irrespective of their stage of liver disease or virus replication rate. Even after successful antiviral therapy fatigue persists in about one third of the patients. Many patients, in addition, report of deficits in attention, concentration and memory, some also of depression. Psychometric testing revealed deficits in attention and verbal learning ability as characteristic for HCV afflicted patients with normal liver function. Magnetic resonance spectroscopic studies showed alterations of the cerebral choline, N-acetyl-aspartate, and creatine content in the basal ganglia, white matter and frontal cortex, respectively. Recently, pathologic cerebral serotonin and dopamine transporter binding and regional alterations of the cerebral glucose utilisation compatible with alterations of the dopaminergic attentional system were observed. Several studies detected HCV in brain samples or cerebro-spinal fluid. Interestingly, viral sequences in the brain often differed from those in the liver, but were closely related to those found in lymphoid tissue. Therefore, the Trojan horse hypothesis emerged: HCV-infected mononuclear blood cells enter the brain, enabling the virus to reside within the brain (probably in microglia) and to infect brain cells, especially astrocytes. PMID- 19130197 TI - Control of oscillation periods and phase durations in half-center central pattern generators: a comparative mechanistic analysis. AB - Central pattern generators (CPGs) consisting of interacting groups of neurons drive a variety of repetitive, rhythmic behaviors in invertebrates and vertebrates, such as arise in locomotion, respiration, mastication, scratching, and so on. These CPGs are able to generate rhythmic activity in the absence of afferent feedback or rhythmic inputs. However, functionally relevant CPGs must adaptively respond to changing demands, manifested as changes in oscillation period or in relative phase durations in response to variations in non-patterned inputs or drives. Although many half-center CPG models, composed of symmetric units linked by reciprocal inhibition yet varying in their intrinsic cellular properties, have been proposed, the precise oscillatory mechanisms operating in most biological CPGs remain unknown. Using numerical simulations and phase-plane analysis, we comparatively investigated how the intrinsic cellular features incorporated in different CPG models, such as subthreshold activation based on a slowly inactivating persistent sodium current, adaptation based on slowly activating calcium-dependent potassium current, or post-inhibitory rebound excitation, can contribute to the control of oscillation period and phase durations in response to changes in excitatory external drive to one or both half centers. Our analysis shows that both the sensitivity of oscillation period to alterations of excitatory drive and the degree to which the duration of each phase can be separately controlled depend strongly on the intrinsic cellular mechanisms involved in rhythm generation and phase transitions. In particular, the CPG formed from units incorporating a slowly inactivating persistent sodium current shows the greatest range of oscillation periods and the greatest degree of independence in phase duration control by asymmetric inputs. These results are explained based on geometric analysis of the phase plane structures corresponding to the dynamics for each CPG type, which in particular helps pinpoint the roles of escape and release from synaptic inhibition in the effects we find. PMID- 19130198 TI - Pc4, a putative movement protein of Rice stripe virus, interacts with a type I DnaJ protein and a small Hsp of rice. AB - Rice stripe virus (RSV) infects rice and causes great yield reduction in some Asian countries. In this study, rice cDNA library was screened by a Gal4-based yeast two-hybrid system using pc4, a putative movement protein of RSV, as the bait. A number of positive colonies were identified and sequence analysis revealed that they might correspond to ten independent proteins. Two of them were selected and further characterized. The two proteins were a J protein and a small Hsp, respectively. Interactions between Pc4 and the two proteins were confirmed using coimmunoprecipitation. Implications of the findings that pc4 interacted with two chaperone proteins were discussed. PMID- 19130199 TI - Complete genome analysis and virulence characteristics of the Louisiana West Nile virus strain LSU-AR01. AB - West Nile virus (WNV) is a member of the Flaviriridae family, which can cause significant morbidity and mortality in birds, horses, and humans. The WNV-LSU AR01 strain was isolated from a dead blue jay in Louisiana in 2001. Phylogenetic analysis using 75 full WNV genomes revealed that the LSU-AR01 strain belongs to a distinct subclade among the North American strains. The LSU-AR01 strain differed from the NY-99 prototypic strain by 26 nucleotides causing six amino acid changes. An asparagine-to-lysine change was located immediately proximal to a known CD8(+)T cell epitope in NS4B, while a glutamine-to-lysine change was located within a predicted CD8(+)T cell epitope in NS5. The LSU-AR01 strain caused pronounced neuronal necrosis, perivascular cuffing and gliosis in comparison to the NY-99-infected mice. These results suggest that the previously identified Connecticut strains may contain highly neurovirulent strains such as the LSU-AR01 that have spread in North America. PMID- 19130200 TI - Transcriptional suppression of IE180 and TK promoters by the EP0 of pseudorabies virus strains Ea and Fa. AB - In our transient expression assays, the IE180 and TK promoters were markedly suppressed by the EP0 of the pseudorabies virus strains Ea and Fa (EP0/Ea, EP0/Fa). This is in contrast with the transactivating activity of EP0 from strain YS-81 (EP0/YS-81) in previous studies. Amino acid sequence alignment revealed several mutations in both EP0/Ea and EP0/Fa compared with EP0/YS-81. Most remarkable is a two-amino acid substitution next to the RING finger domain which was considered to be important for the transactivating activity of EP0 in previous studies. To address the effect of the two-amino acid substitution on the function of EP0, reverse mutants of EP0/Ea and EP0/Fa were generated. The subsequent expression assays indicated that this substitution was at least in part responsible for the effect on the regulatory activity of EP0. Our data suggested that EP0 may regulate the expression of the same genes in different pseudorabies virus strains differently. PMID- 19130201 TI - Genetic variation in the long terminal repeat associated with the transition of Chinese equine infectious anemia virus from virulence to avirulence. AB - A highly virulent strain of equine infectious anemia virus (EIAV) lost its fatal virulence but retained the desired antigens during serial passage over 130 generations in leukocytes in vitro. We compared the long terminal repeat (LTR) sequences of the different generations and found that three stable genetic variations occurred in the transcriptional start site, the initial base of TAR, and the pre-mRNA cleavage site at the R-U5 boundary, respectively. These three mutations happened at the inflexion of virus pathogenicity loss; therefore, the function of these mutations needs to be further addressed. PMID- 19130202 TI - Molecular analysis of US10, S3, and US2 in duck enteritis virus. AB - A 4554-bp fragment was amplified from the DEV C-KCE vaccine strain by single oligonucleotide nested polymerase chain reaction with partially known sequences for the DEV US1 and US10 genes. Three open reading frames containing the genes encoding US10, S3, and US2 were predicted using the Editseq program (DNAStar). The S3 and US2 genes have the same transcription orientation but are oriented head-to-head with respect to US10. The promoters and polyadenylation signals were predicted. Two poly A sequences were predicted in S3, but none were predicted in US2. These results provide partial sequence of US region for the physical map of the DEV genome. Phylogenetic analysis suggests that the DEV C-KCE strain is more closely related to Mardivirus in the alphaherpesvirus subfamily of the Herpesviridae. PMID- 19130204 TI - Sexual narcissism and the perpetration of sexual aggression. AB - Despite indirect evidence linking narcissism to sexual aggression, studies directly examining this relationship have yielded inconsistent results. Likely contributing to such inconsistencies, prior research has used global measures of narcissism not sensitive to whether the components of narcissism are activated in sexual versus non-sexual domains. The current research avoided such problems by using a measure of sexual narcissism to predict sexual aggression. In a sample of 299 men and women, Study 1 validated the Sexual Narcissism Scale, a new sexuality research instrument with four subscales-Sexual Exploitation, Sexual Entitlement, Low Sexual Empathy, and Sexual Skill. Then, in a sample of 378 men, Study 2 demonstrated that sexual narcissism was associated with reports of the frequency of sexual aggression, three specific types of sexual aggression (unwanted sexual contact, sexual coercion, and attempted/completed rape), and the likelihood of future sexual aggression. Notably, global narcissism was unrelated to all indices of sexual aggression when sexual narcissism was controlled. That sexual narcissism outperformed global assessments of narcissism to account for variance in sexual aggression suggests that future research may benefit by examining whether sexual narcissism and other sexual-situation-specific measurements of personality can similarly provide a more valid test of the association between personality and other sexual behaviors and outcomes (e.g., contraceptive use, infidelity, sexual satisfaction). PMID- 19130205 TI - Sex differences in mental rotation and line angle judgments are positively associated with gender equality and economic development across 53 nations. AB - Mental rotation and line angle judgment performance were assessed in more than 90,000 women and 111,000 men from 53 nations. In all nations, men's mean performance exceeded women's on these two visuospatial tasks. Gender equality (as assessed by United Nations indices) and economic development (as assessed by per capita income and life expectancy) were significantly associated, across nations, with larger sex differences, contrary to the predictions of social role theory. For both men and women, across nations, gender equality and economic development were significantly associated with better performance on the two visuospatial tasks. However, these associations were stronger for the mental rotation task than for the line angle judgment task, and they were stronger for men than for women. Results were discussed in terms of evolutionary, social role, and stereotype threat theories of sex differences. PMID- 19130206 TI - Does physical abuse, sexual abuse, or neglect in childhood increase the likelihood of same-sex sexual relationships and cohabitation? A prospective 30 year follow-up. AB - Existing cross-sectional research suggests associations between physical and sexual abuse in childhood and same-sex sexual orientation in adulthood. This study prospectively examined whether abuse and/or neglect in childhood were associated with increased likelihood of same-sex partnerships in adulthood. The sample included physically abused (N = 85), sexually abused (N = 72), and neglected (N = 429) children (ages 0-11) with documented cases during 1967-1971 who were matched with non-maltreated children (N = 415) and followed into adulthood. At approximately age 40, participants (483 women and 461 men) were asked about romantic cohabitation and sexual partners, in the context of in person interviews covering a range of topics. Group (abuse/neglect versus control) differences were assessed with cross-tabulations and logistic regression. A total of 8% of the overall sample reported any same-sex relationship (cohabitation or sexual partners). Childhood physical abuse and neglect were not significantly associated with same-sex cohabitation or sexual partners. Individuals with documented histories of childhood sexual abuse were significantly more likely than controls to report ever having had same-sex sexual partners (OR = 2.81, 95% CI = 1.16-6.80, p < or = .05); however, only men with histories of childhood sexual abuse were significantly more likely than controls to report same-sex sexual partners (OR = 6.75, 95% CI = 1.53-29.86, p < or = .01). These prospective findings provide tentative evidence of a link between childhood sexual abuse and same-sex sexual partnerships among men, although further research is needed to explore this relationship and to examine potential underlying mechanisms. PMID- 19130207 TI - Predictors and consequences of sexual "hookups" among college students: a short term prospective study. AB - "Hookups," sexual interactions between partners who do not expect a romantic commitment, are believed to be common among adolescents and young adults. Most existing research is cross-sectional and has not investigated the antecedents or consequences of hookups. To our knowledge, this study provides the first prospective investigation of the hypothesized predictors of penetrative sex hookups (i.e., oral, vaginal, and anal sex) and the first exploration of the short-term mental health consequences of hookups. A total of 140 first-semester college students (109 females, 31 males) completed an anonymous survey early in their first semester; the survey assessed 18 potential predictors of hooking up that were identified from theory and past research. At the end of their first semester, students again responded to a survey and provided data on their oral and vaginal sex hookup behavior (occurrence and number of partners), distress, and self-esteem. Baseline and follow-up data were linked using unique codes that protected participants' anonymity. Pre-college hookup patterns, peak intoxication level, and situational triggers for hookups were consistent predictors of oral and vaginal sex hookup behavior (and number of hookup partners) in the first semester of college. Penetrative sex hookups increased psychological distress for females, but not for males. Implications for education and intervention as well as suggestions for future research are discussed. PMID- 19130208 TI - Menstrual cycle phase effects on memory and Stroop task performance. AB - The present study examined differences in Stroop and memory task performances modulated by gonadal steroid hormones during the menstrual cycle in women. Thirty women with regular menstrual cycles performed a logical memory task (Wechsler Memory Scale) and the Stroop task. The results showed a significant difference in Stroop task performance between low and high levels of estradiol and progesterone during the menstrual cycle, but there was no significant difference in memory performance between the two phases, nor was there any significant mood change that might have influenced cognitive performance. These findings suggest that sex related hormone modulation selectively affects cognitive functions depending on the type of task and low level secretion of estradiol appears to contribute to reducing the level of attention that relates to the prefrontal cortex. PMID- 19130209 TI - A genome wide detection of quantitative trait loci on pig maternal infanticide behavior in a large scale White Duroc x Erhualian resource population. AB - Maternal behavior around parturition is important to piglet survival. An extreme form of failure of maternal behavior, also called maternal infanticide, often occurs in some sows. This is defined as an active attack to piglets using the jaws, resulting in serious or fatal bite wounds within 24 h of birth. It leads to considerable economic losses to the pig industry and severe problems in pig welfare. In this study, maternal behaviors from 5 h before to 24 h after parturition were recorded in detail on 288 White Duroc x Erhualian intercross F(2) sows over their three continuous farrowings. In the F(2) population 12.8% gilts showed maternal infanticide in their first litter, while the incidences of maternal infanticide at their second and third farrowing reduced to 7.5% and 4.5%, respectively. All F(2) sows were genotyped for 194 microsatellite markers spanning the whole pig genome. A whole genome linkage analysis was performed using the non-parametric linkage test by SimWalk2 software. The results identified that seven chromosome regions on SSC2, SSC6, SSC14, SSC15 and SSCX were significantly linked with maternal infanticide (P < 0.05). The quantitative trait loci (QTL) on SSC2 and SSCX achieved P < 0.01 significance level. The most promising QTLs, however, were detected on X chromosome where three peaks of negative logarithm of P-value located at marker SW980, SW2456 and SW1608. QTLs on SSC2 and SSCX from this experiment were consistent with published results from the Western commercial lines. PMID- 19130210 TI - Why a special section about community psychology and politics. PMID- 19130211 TI - Community action and research as citizenship construction. AB - Social change, well-being and liberation have been intertwined in community research and action, as much as being used as political common-places. In this paper, it is argued that community research and action can have a political character. The epistemological premises shared by both the community and political spheres are discussed, and concepts are defined. The way community psychological action can produce changes is illustrated by an experience carried out with children from poverty sectors of Caracas, Venezuela, intended to construct citizenship by developing awareness about values, civic rights and duties, while experiencing the advantages of participation, peaceful negotiation and solidarity, during art classes. Topics regarding citizenship values and ethics were painted and discussed in a participatory way. Results about how the children developed consciousness about those topics, relating them to their lives, are presented. PMID- 19130212 TI - Cultural predictors of physical and mental health status among Mexican American women: a mediation model. AB - This study, using secondary data analysis, examined a mediation model of acculturation and ethnic pride as predictors of physical and mental health outcomes in a sample of 561 Mexican American women. Factors postulated as mediators were family support and religiosity. Systematic across-group comparison analyses were conducted to examine sources of differences in the mediation model between immigrant and non-immigrant women. The results partially supported the hypothesized mediation model, indicating that family support, but not religiosity, was a significant mediator in the relationship between ethnic pride and mental health problems. In addition, as differences between immigrant and non immigrant women were observed only in the variables means, but not in the factor loadings or regression paths, the model tested may capture a common psychosocial process that affects these women and their health outcomes. Overall, this study offers important implications for future research and the design of intervention programs for Mexican American women. PMID- 19130213 TI - Income volatility and psychological depression. AB - Income volatility appears to be increasing especially among lower income workers. Such volatility may reflect the ongoing shift of economic risk from employers to employees as marked by decreasing job security and employer-provided benefits. This study tests whether absolute volatility or downward volatility in income predict depression controlling for prior depression. A sample (n = 4,493) from the National Longitudinal Survey of Youth (NLSY79) with depression (CESD) measured at age 40 and prior depression measured eight to 10 years earlier was utilized. Downward volatility (frequency of income loss) was positively associated with depression; adjusting for downward volatility and other covariates, absolute volatility was negatively associated with depression. An interaction indicated a positive association between downward volatility and depression only when absolute volatility was high. These findings apply to respondents in a narrow age range (30 s) and the results warrant replication to identify the mediators linking absolute volatility and income loss to depression. PMID- 19130214 TI - Globalization, neo-liberalism and community psychology. AB - A longitudinal analysis (1984-2005) of media language in Norway is presented, demonstrating how the current globalized capitalist market ideology is now permeating this long-established Scandinavian welfare state. This ideological shift carries powerful implications for community psychology, as traditional welfare state values of equal services based on a universalistic principle are set aside, and social and material inequalities are increasingly accepted. The methodology developed in the present study may serve as a "barometer of community changes", to borrow a metaphor used by Sarason (2000). PMID- 19130215 TI - 17beta-estradiol induces transthyretin expression in murine choroid plexus via an oestrogen receptor dependent pathway. AB - Oestrogen protects against AD by multiple mechanisms, including the enhancement of Abeta clearance. Transthyretin (TTR) is a homotetrameric protein mainly synthesized by the liver and choroid plexus (CP) of the brain that sequesters the amyloid beta (Abeta) peptide. In this study we examined the effects of 17beta estradiol (E2) on TTR protein and mRNA levels, in primary cultures of rat CP epithelial cells (CPEC) by Western blot and Real Time PCR, respectively. Moreover, the localization of oestrogen receptors alpha (ERalpha) and beta (ERbeta) in response to E2 treatment was analysed by confocal microscopy in these cells. The expression of TTR, ERalpha and ERbeta was also compared in the CP of castrated female mice treated with E2 to vehicle-treated animals by Real Time PCR. TTR concentration in the CSF of all these animals was measured by radioimmunoassay. E2 treatment induced TTR transcription and increased TTR protein content in CPEC. Pre-treatment with ICI 182,780 (ICI) abrogated E2 induced TTR expression suggesting that, TTR is up-regulated via an ER-dependent pathway. Confocal microscopy demonstrated extranuclear ERalpha and ERbeta localization in untreated CPEC. Upon E2 treatment, translocation of ERalpha to the nucleus occurred, while ERbeta remained in the cytosol. These data was concurrent with the up-regulation of TTR expression detected in the CP of castrated female mice subjected to E2 treatment. Our results highlight the importance of E2 on the regulation of TTR, which may participate in the oestrogen induced decrease in Abeta levels and deposition described in the literature. PMID- 19130216 TI - Gene expression profiling of human neural progenitor cells following the serum induced astrocyte differentiation. AB - Neural stem cells (NSC) with self-renewal and multipotent properties could provide an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. However, the majority of transplanted NSC and neural progenitor cells (NPC) differentiate into astrocytes in vivo under pathological environments in the central nervous system, which potentially cause reactive gliosis. Because the serum is a potent inducer of astrocyte differentiation of rodent NPC in culture, we studied the effect of the serum on gene expression profile of cultured human NPC to identify the gene signature of astrocyte differentiation of human NPC. Human NPC spheres maintained in the serum free culture medium were exposed to 10% fetal bovine serum (FBS) for 72 h, and processed for analyzing on a Whole Human Genome Microarray of 41,000 genes, and the microarray data were validated by real-time RT-PCR. The serum elevated the levels of expression of 45 genes, including ID1, ID2, ID3, CTGF, TGFA, METRN, GFAP, CRYAB and CSPG3, whereas it reduced the expression of 23 genes, such as DLL1, DLL3, PDGFRA, SOX4, CSPG4, GAS1 and HES5. Thus, the serum-induced astrocyte differentiation of human NPC is characterized by a counteraction of ID family genes on Delta family genes. Coimmunoprecipitation analysis identified ID1 as a direct binding partner of a proneural basic helix-loop-helix (bHLH) transcription factor MASH1. Luciferase assay indicated that activation of the DLL1 promoter by MASH1 was counteracted by ID1. Bone morphogenetic protein 4 (BMP4) elevated the levels of ID1 and GFAP expression in NPC under the serum-free culture conditions. Because the serum contains BMP4, these results suggest that the serum factor(s), most probably BMP4, induces astrocyte differentiation by upregulating the expression of ID family genes that repress the proneural bHLH protein-mediated Delta expression in human NPC. PMID- 19130217 TI - De-differentiation response of cultured astrocytes to injury induced by scratch or conditioned culture medium of scratch-insulted astrocytes. AB - Our previous reports indicated that astrocytes (ASTs) in injured adult rat spinal cord underwent a process of de-differentiation, and may acquire the potential of neural stem cells (NSCs). However, the AST de-differentiation and transitional rejuvenation process following injury is still largely unclear. The aim of the present study was to determine whether injured in vitro ASTs can re-enter the multipotential-like stem cell pool and regain NSC characteristics, and to further understand the mechanism of AST de-differentiation. We used an in vitro scratch wound model to evoke astrocytic response to mechanical injury. GFAP and nestin double-labeled indirect immunofluorescence were carried out to characterize these scratched cells at various periods. Western-blot analysis was used to determine the changes of GFAP and nestin expression following injury. Furthermore, the rate of proliferation was determined by immunocytochemical detection of BrdU incorporating cells. These scratch-wound ASTs were cultured with stem cells medium to explore their ability to generate neurospheres and examine the self renewal and multi-potency of such neurospheres. Moreover, scratched AST culture supernatant as conditioned cultured medium (ACM) was used to investigate if some diffusible factors derived from injured ASTs could induce de-differentiation of AST. The results showed: (1) the nestin positivity first appeared in GFAP positive cells at the edge of the scratch, subsequently, disseminated into un insulted zone. The expression of nestin in AST was increased with longer culture, while that of GFAP was decreased. Furthermore, these nestin-immunoreactive ASTs could generate neurospheres, which showed self-renewal and could be differentiated into neurons, ASTs and oligodendrocytes. (2) Scratched ASTs culture supernatant can induce astrocytic proliferation and de-differentiation. These results reveal that the in vitro injured ASTs can de-differentiate into nestin-positive stem/precursor cells, the process of de-differentiation may arise from direct injury or some diffusible factors released from injured ASTs. PMID- 19130218 TI - Cost-utility analysis for advanced breast cancer therapy in Germany: results of the fulvestrant sequencing model. AB - Therapy decisions in advanced breast cancer (ABC) increasingly require assessment not only of treatment efficacy but also of cost-effectiveness. To this end, we performed a cost-utility analysis by comparing treatment sequences including/omitting fulvestrant in a hypothetical population of hormone receptor positive (HR+) postmenopausal women with ABC. The analysis was performed from the German health care perspective. Using a first-order sequential Markov model, expected costs and utilities were calculated over a time horizon of 10 years for cohorts of patients with HR+ ABC, previously treated for at least 5 years using adjuvant endocrine therapies. Utilities were primarily quantified in terms of quality adjusted life years (QALY). "Base-case" estimates of state transition rates, resource utilization, and other model parameters were derived from published evidence and expert assessment. The impacts of uncertainties in all key model parameters were evaluated by sensitivity analysis. Costs and benefits were discounted at 3% annually. Including second-line fulvestrant in the treatment sequence led to greater estimated health gains (0.021 QALY) and cost savings of 564 euros ($745, 380 pounds) per patient, i.e. the fulvestrant-containing sequence was "dominant". The prediction of a cost savings was robust with respect to variations in all key parameters. The probability of acceptable cost effectiveness for the fulvestrant sequence was 72% at a willingness to pay (WTP) of 30,000 euros/QALY ($39,621/QALY, 20,198 pounds/QALY); the probability was even higher at lower WTP and substantially exceeded 50% for any realistic WTP. In a representative population of women with HR+ advanced breast cancer, inclusion of fulvestrant in the treatment sequence provides a cost-effective alternative from the German health care perspective. A high probability of cost-effectiveness is maintained under variations in all key parameters. The results reflect a tendency for patients receiving fulvestrant at an early stage to maintain high quality of life for a longer interval. PMID- 19130219 TI - Risk factors for brain relapse in HER2-positive metastatic breast cancer patients. AB - Brain relapse is a common occurrence in HER2-positive breast cancer patients. However, the factors determining the risk of brain metastasis in these patients remain to be established. The aim of this study was to assess the impact of particular clinical and pathological factors on the risk of brain relapse in HER2 positive advanced breast cancer patients. The study group included 264 consecutive HER-2 positive metastatic breast cancer patients, most of whom (210; 80%) were administered trastuzumab, usually in combination with chemotherapy. Time from the diagnosis to distant relapse ranged from 0 to 142 months (median 16 months). The most common dominant site of metastatic disease was viscera (80%), followed by soft tissue (11%) and bones (10%). After a median follow-up of 3.1 years, the symptomatic brain relapse occurred in 103 patients (39%). Median time from treatment dissemination to brain relapse was 15 months (range, 0-81 months), and the cumulative 1-year, 3-year and 5-year risk of brain relapse was 17, 42 and 55%, respectively. The average annual risk of brain relapse for surviving patients during consecutive 7 years of follow-up was 10.0% (95% CI, 6.6-13.5%). In the univariate analysis the only variable significantly related to the increased risk of brain relapse was time from initial diagnosis to distant relapse shorter than 2 years (HR = 1.55, 95% CI, 1.03-2.33, P = 0.034). Patients with dominant site of disease in soft tissue or bones tended to have lower risk of relapse (HR = 0.54 and 0.62; P = 0.098 and 0.203, respectively) compared to patients with visceral metastases. Treatment with trastuzumab was not associated with reduced risk of brain relapse (HR = 0.91, 95% CI, 0.47-1.77, P = 0.78). In the multivariate analysis, time from initial diagnosis to distant relapse shorter than 2 years remained the only significant variable related to increased risk of brain relapse (adjusted HR = 1.62, 95% CI, 1.07-2.44; P = 0.022). HER2-positive breast cancer patients remain at high and continuous risk of brain relapse for a prolonged period of time after diagnosis of disease dissemination. Short time from initial diagnosis to distant relapse is related to increased risk of brain relapse. Molecular predictors are sorely needed to better characterize patients with high probability of early brain relapse. PMID- 19130220 TI - Generic script share and the price of brand-name drugs: the role of consumer choice. AB - Pharmaceutical expenditures have grown rapidly in recent decades, and now total nearly 10% of health care costs. Generic drug utilization has risen substantially alongside, from 19% of scripts in 1984 to 47% in 2001, thus tempering expenditure growth through significant direct dollar savings. However, generic drugs may lead to indirect savings as well if their use reduces the average price of those brand name drugs that are still purchased. Prior work indicates that brand-name producers do not lower their prices in the face of generic competition, and our study confirms that finding. However, prior work is silent on how the mix of consumer choices between generic and brand-name drugs might affect the average price of those brand-name drugs that are purchased. We use a nationally representative panel of data on drug utilization and costs for the years 1996 2001 to examine how the share of an individual's prescriptions filled by generics (generic script share) affects his average out-of-pocket cost for brand-name drugs, and the net cost paid by the insurer. Our principal finding is that a higher generic script share lowers average brand-name prices to consumers, presumably because consumers are more likely to substitute generics when brand name drugs would cost them more. This effect is substantial: a 10% increase in the consumer's generic script share is associated with a 15.6% decline in the average price paid for brand-name drugs by consumers. This implies that the potential cost savings to consumers from generic substitution are far greater than prior work suggests. In contrast, the percentage reduction in average brand costs to health plans is far smaller, and statistically insignificant. PMID- 19130222 TI - Transitioning clients from assertive community treatment to traditional mental health services. AB - The original assertive community treatment (ACT) model included time-unlimited treatment. However, resource restrictions and subsequent research call into question the feasibility and necessity of time-unlimited services. This study compares the outcomes of 48 individuals who successfully transitioned from ACT to less intensive Community Mental Health Center (CMHC) services to 19 persons who required return to ACT or did not remain in CMHC treatment. There were no statistically significant differences in demographics, diagnoses, and other treatment factors between the two groups. PMID- 19130221 TI - Insulin action on polyunsaturated phosphatidic acid formation in rat brain: an "in vitro" model with synaptic endings from cerebral cortex and hippocampus. AB - The highly efficient formation of phosphatidic acid from exogenous 1-stearoyl-2 arachidonoyl-sn-glycerol (SAG) in rat brain synaptic nerve endings (synaptosomes) from cerebral cortex and hippocampus is reported. Phosphatidic acid synthesized from SAG or 1,2-dipalmitoyl-sn-glycerol (DPG) was 17.5 or 2.5 times higher, respectively, than from endogenous synaptosomal diacylglycerides. Insulin increased diacylglycerol kinase (DAGK) action on endogenous substrate in synaptic terminals from hippocampus and cerebral cortex by 199 and 97%, respectively. Insulin preferentially increased SAG phosphorylation from hippocampal membranes. In CC synaptosomes insulin increased phosphatidic acid (PA) synthesis from SAG by 100% with respect to controls. Genistein (a tyrosine kinase inhibitor) inhibited this stimulatory insulin effect. Okadaic acid or cyclosporine, used as Ser/Threo protein phosphatase inhibitors, failed to increase insulin effect on PA formation. GTP gamma S and particularly NaF were potent stimulators of PA formation from polyunsaturated diacylglycerol but failed to increase this phosphorylation when added after 5 min of insulin exposure. GTP gamma S and NaF increased phosphatidylinositol 4,5 bisphosphate (PIP2) labeling with respect to controls when SAG was present. On the contrary, they decreased polyphosphoinositide labeling with respect to controls in the presence of DPG. Our results indicate that a DAGK type 3 (DAGKepsilon) which preferentially, but not selectively, utilizes 1-acyl-2-arachidonoyl-sn-glycerol and which could be associated with polyphosphoinositide resynthesis, participates in synaptic insulin signaling. GTP gamma S and NaF appear to be G protein activators related to insulin and the insulin receptor, both affecting the signaling mechanism that augments phosphatidic acid formation. PMID- 19130223 TI - Noninvasive monitoring of transient cardiac changes with impedance cardiography. AB - Impedance cardiography (IC) has been used to monitor the pumping function of the heart in terms of stroke volume or cardiac output. We explored the extent to which IC can be used for monitoring transient effects of cardiac alterations on a beat-to-beat basis. A four-spot-electrode configuration was used in the present design. The spatial arrangement of the four electrodes was carefully selected to obtain an optimal result. A constant current source provided a 4 mA, 20 kHz sinusoidal excitation current. Analog signal processing was developed to demodulate and amplify the resultant impedance information, eliminate low frequency motion artifacts and incidental breathing artifacts. Experiments were performed on normal healthy volunteers. Impedance waveforms were recorded from each subject beginning from a supine position then moving to an upright position. Breath holding was maintained throughout the procedure. The impedance waveform, Z, its first derivative dZ/dt, the DC component, Z(0), and electrocardiogram were recorded simultaneously and continuously. The results showed a significant increase in the relative impedance levels from supine to transient state and then decreased during the upright position. Additionally, the supine position had a higher (dZ/dt)(max) than sitting resulting in a reduction in stroke volume. The increased impedance from supine to sitting verified the physiological findings obtained by others. The results of this investigation illustrate the ability of impedance cardiography to effectively monitor the dynamic cardiovascular function under transient conditions. PMID- 19130225 TI - An exploratory analysis of healthcare costs and utilization of pediatric patients with Crohn's disease. AB - OBJECTIVE: To evaluate the healthcare costs and resource utilization associated with pediatric Crohn's disease (PCD) from a payer perspective. METHODS: A retrospective analysis was conducted using claims from 1 January 2003 through 31 December 2006 from the HealthCare Partners database. Patients were younger than 18 years of age, had a new diagnosis of PCD, and continuous health plan eligibility 6 months before and 12 months after the disease index date (the date of the first claim with a PCD diagnosis). For comparison, a non-PCD cohort was matched on age, sex, and birthday (within 30 days). RESULTS: Data from 30 patients with PCD and 10,864 non-PCD controls were included. The total cost per member per month (PMPM) for the PCD cohort was $2,547 compared with $101 for the non-PCD cohort. Inpatient admissions accounted for the largest portion (54%) of the total cost PMPM for PCD patients. There were 500 admissions per thousand members per year (PTMPY) for the PCD cohort and 11 admissions PTMPY for the non PCD cohort. The average lengths of stay were 7.6 and 4.4 days for the PCD cohort and the non-PCD cohort, respectively, and the inpatient costs PMPM were $1,409 and $18, respectively. Costs and resource utilization were also higher for PCD patients treated with systemic therapies. CONCLUSION: PCD was associated with higher costs and resource utilization, compared with non-PCD controls, primarily driven by inpatient stays. Treating PCD appropriately before the disease progresses to a level requiring hospitalization may help reduce the costs associated with this disease. PMID- 19130224 TI - Gastroprotective action of Cochinchina momordica seed extract is mediated by activation of CGRP and inhibition of cPLA(2)/5-LOX pathway. AB - Cochinchina momordica seed extract (SKMS10), which is composed of the major compounds momordica saponins, has been evaluated for its gastroprotective effects in rat models of acute gastric mucosal damage. Ethanol and water immersion restraint stress (WRS) induced gastric damage, including hemorrhages and edema, was significantly attenuated by pretreatment with SK-MS10. In addition, SK-MS10 reduced increases of mucosal myeloperoxidase (MPO), IL-1beta, and TNFalpha levels and the expression of cPLA(2), and 5-LOX induced by ethanol or WRS. SK-MS10 also increased hexosamine, adherent mucus, and the expression of MUC5AC. Furthermore, SK-MS10 enhanced the mucosal expression of the CGRP gene and its serum levels.N(G)-methyl L-arginine (L-NMMA) or capsaicin desensitization reversed the SK-MS10-induced gastroprotection effect. These results suggest that SK-MS10 is a gastroprotective agent against acute gastric mucosal damage by suppressing proinflammatory cytokines, downregulating cPLA(2), 5-LOX, and increasing the synthesis of mucus. Furthermore, CGRP-NO pathway was found to play an important role in these gastroprotective effects of SK-MS10. PMID- 19130227 TI - Analysis of MEG background activity in Alzheimer's disease using nonlinear methods and ANFIS. AB - This study was designed to analyze the magnetoencephalogram (MEG) background activity from 20 patients with probable Alzheimer's disease (AD) and 21 control subjects by using two nonlinear methods: sample entropy (SampEn), and Lempel-Ziv complexity (LZC). The former quantifies the signal regularity, and the latter is a complexity measure. The signals were acquired with a 148-channel whole-head magnetometer placed in a magnetically shielded room. Our results show that MEG recordings are less complex and more regular in patients with AD than in control subjects. Significant differences between both groups were found in 16 MEG channels with SampEn and in 134 with LZC (p < 0.01, Student's t test with Bonferroni's correction). Using receiver operating characteristic curves with a leave-one-out cross-validation procedure, accuracies of 70.73 and 78.05% were reached with SampEn and LZC, respectively. Additionally, we wanted to assess whether both nonlinear methods and an adaptive-network-based fuzzy interference system (ANFIS) could improve AD diagnosis. With this classifier, an accuracy of 85.37% was achieved. Our findings suggest the usefulness of our methodology to increase our insight into AD. PMID- 19130228 TI - Effect of low-frequency pulsatile flow on expression of osteoblastic genes by bone marrow stromal cells. AB - Perfusion culture of osteoprogenitor cells is a promising means to form a bone like extracellular matrix for tissue engineering applications, but the mechanism by which hydrodynamic shear stimulates expression of bone extracellular matrix (ECM) proteins is not understood. Osteoblasts are mechanosensitive and respond differently to steady and pulsatile flow. Therefore, to probe the effect of flow, bone marrow stromal cells (BMSCs)--cultured under osteogenic conditions--were exposed to steady or pulsatile flow at frequencies of 0.015, 0.044, or 0.074 Hz. Following 24 h of stimulus, cells were cultured statically for an additional 13 days and then analyzed for the expression of bone ECM proteins collagen 1alpha1 (Col1alpha1), osteopontin, osteocalcin (OC), and bone sialoprotein (BSP). All mRNA levels were elevated by flow, but OC and BSP were enhanced modestly with pulsatile flow. To determine if these effects were related to gene induction during flow, BMSCs were again exposed to steady or pulsatile flow for 24 h, but then analyzed immediately for expression of growth and differentiation factors bone morphogenetic proteins (BMP)-2, -4, and -7, transforming growth factor (TGF) beta1, and vascular endothelial growth factor-A. All growth and differentiation factors were significantly elevated by flow, except BMP-4 which was suppressed. In addition, expression of BMP-2 and -7 were enhanced and TGF-beta1 suppressed by pulsatile flow relative to steady flow. These results demonstrate that pulsatile flow modulates expression of BMP-2, -7, and TGF-beta1 and suggest that enhanced expression of bone ECM proteins by pulsatile flow may be mediated through the induction of BMP-2 and -7. PMID- 19130226 TI - Inflammatory bowel disease: a patient's and caregiver's perspective. AB - The purpose of this study was to conduct a survey examining the impact of inflammatory bowel disease (IBD) on patients' and their caregivers' daily activities. Questionnaires were distributed to patients registered in the APDI (Portuguese Association for IBD) database and their respective caregivers in 2007. Of 422 patient respondents, 251 had Crohn's disease (CD) and 171 had ulcerative colitis (UC), with the majority of patients being women (58.1%) and aged over 40 years (37.4%). The number of disease flares experienced by IBD patients was slightly higher for patients with CD than for patients with UC (2.64 vs. 2.34), and surgery was more often required in CD patients as compared to UC patients (42.4 vs. 7%). Sixty percent (60%) of patients reported having no problems with mobility, daily activities, or personal hygiene; however, over half of all patients experienced some pain and anxiety. Adult patients and children and adolescents respectively experienced time off work or school due to their disease but caregivers were not affected in this regard. The caregivers life (N=324) was affected by anxiety, with the major concern reported as the risk of the patient developing cancer. Both IBD patients and caregivers thought that the provision of information on new drugs and contact time with a doctor would have the biggest impact on improving care. The symptoms and complications of IBD have a considerable impact on the lives of patients and their caregivers, and several actions could be taken to improve their care. PMID- 19130229 TI - MRI-based CFD analysis of flow in a human left ventricle: methodology and application to a healthy heart. AB - A three-dimensional computational fluid dynamics (CFD) method has been developed to simulate the flow in a pumping left ventricle. The proposed method uses magnetic resonance imaging (MRI) technology to provide a patient specific, time dependent geometry of the ventricle to be simulated. Standard clinical imaging procedures were used in this study. A two-dimensional time-dependent orifice representation of the heart valves was used. The location and size of the valves is estimated based on additional long axis images through the valves. A semi automatic grid generator was created to generate the calculation grid. Since the time resolution of the MR scans does not fit the requirements of the CFD calculations a third order bezier approximation scheme was developed to realize a smooth wall boundary and grid movement. The calculation was performed by a Navier Stokes solver using the arbitrary Lagrange-Euler (ALE) formulation. Results show that during diastole, blood flow through the mitral valve forms an asymmetric jet, leading to an asymmetric development of the initial vortex ring. These flow features are in reasonable agreement with in vivo measurements but also show an extremely high sensitivity to the boundary conditions imposed at the inflow. Changes in the atrial representation severely alter the resulting flow field. These shortcomings will have to be addressed in further studies, possibly by inclusion of the real atrial geometry, and imply additional requirements for the clinical imaging processes. PMID- 19130230 TI - Cell death in hematological tumors. AB - Evasion of apoptosis is a hallmark of human cancers, for example in hematological malignancies. Apoptosis is an intrinsic cell death program that is crucial to maintain tissue homeostasis, for example in the hematopoietic system where there is a high turnover rate of cells. As a result, a decrease in the rate of apoptosis besides an increase in proliferation favors tumorigenesis as well as tumor progression. Further, the anti-leukemic action of current treatment approaches, including chemo-, radio- or immunotherapy, critically relies on intact cell death programs in cancer cells. Therefore, defects in apoptosis pathways are frequently associated with the resistance to anticancer therapies. In recent years, the identification and characterization of the molecules and pathways that are involved in the regulation and execution of cell death in leukemia and lymphoma cells have set the ground for the development of novel diagnostic tools and molecular therapeutics targeting apoptosis pathways in hematological malignancies. PMID- 19130231 TI - Cell death: protein misfolding and neurodegenerative diseases. AB - Several chronic neurodegenerative disorders manifest deposits of misfolded or aggregated proteins. Genetic mutations are the root cause for protein misfolding in rare families, but the majority of patients have sporadic forms possibly related to environmental factors. In some cases, the ubiquitin-proteasome system or molecular chaperones can prevent accumulation of aberrantly folded proteins. Recent studies suggest that generation of excessive nitric oxide (NO) and reactive oxygen species (ROS), in part due to overactivity of the NMDA-subtype of glutamate receptor, can mediate protein misfolding in the absence of genetic predisposition. S-Nitrosylation, or covalent reaction of NO with specific protein thiol groups, represents one mechanism contributing to NO-induced protein misfolding and neurotoxicity. Here, we present evidence suggesting that NO contributes to protein misfolding via S-nitrosylating protein-disulfide isomerase or the E3 ubiquitin ligase parkin. We discuss how memantine/NitroMemantine can inhibit excessive NMDA receptor activity to ameliorate NO production, protein misfolding, and neurodegeneration. PMID- 19130232 TI - Cell death in allergic diseases. AB - Apoptosis, the most common form of cell death, is a key mechanism in the build up and maintenance of both innate and adaptive immunity. Central to the apoptotic process is a family of intracellular cysteine proteases with aspartate specificity, called caspases. Caspases are counter-regulated by multiple anti apoptotic molecules, and the expression of the latter in leukocytes is largely dependent on survival factors. Therefore, the physiologic rates of apoptosis change under pathologic conditions. For instance, in inflammation, the expression of survival factors is usually elevated, resulting in increased cell survival and consequently in the accumulation of the involved immune cells. In many allergic diseases, eosinophil apoptosis is delayed contributing to both blood and tissue eosinophilia. Besides eosinophils, apoptosis of other leukocytes is also frequently prevented or delayed during allergic inflammatory processes. In contrast to inflammatory cells, accelerated cell death is often observed in epithelial cells, a mechanism, which amplifies or at least maintains allergic inflammation. In conclusion, deregulated cell death is a common phenomenon of allergic diseases that likely plays an important role in their pathogenesis. Whether the apoptosis is too little or too much depends on the cell type. In this review, we discuss the regulation of the lifespan of the participating leukocytes in allergic inflammatory responses. PMID- 19130233 TI - Dietary polyunsaturated fatty acids as inducers of apoptosis: implications for cancer. AB - It has recently become clear the role played by alterations in apoptosis during the development of several chronic diseases (i.e. inflammatory, neurodegenerative and neoplastic pathologies). For this reason, the research for possible therapeutic strategies involving the modulation of the apoptotic pathways has attracted considerable interest in the past few years. In particular, it has been shown that apoptosis may be induced or inhibited by a variety of nutritional compounds providing health benefits. The aim of this review is to examine the ability of different dietary polyunsaturated fatty acids (PUFAs) to induce apoptosis, especially in the cancer field. The molecular effects of different PUFAs found in dairy products, meat, fish, vegetable seeds and oils, and known to affect the incidence and progression of cancer and other chronic diseases, will be analyzed. To this aim, our effort will concentrate in critically reviewing the published works concerning the effects of: (a) the n-6 PUFAs gamma-linolenic acid, arachidonic acid, and conjugated linoleic acid; (b) the n-3 PUFAs eicosapentaenoic acid and docosahexaenoic acid on the apoptotic process. We will also pay attention to the recent findings regarding the possible role of PUFAs as regulators of the endoplasmic reticulum stress-pathway of apoptosis. PMID- 19130234 TI - Pan-caspase inhibition suppresses polyethylene particle-induced osteolysis. AB - Particle-induced osteolysis is a major cause of aseptic loosening after total joint replacement. Earlier studies demonstrated apoptotic macrophages, giant cells, fibroblasts and T-lymphocytes in capsules and interface membranes of patients with aseptic hip implant loosening. The aim of the current study was to determine in a murine calvarial model of wear particle-induced osteolysis whether inhibition of apoptosis using the pan-caspase inhibitor BOC-D-FMK reduces aseptic loosening. Healthy 12-week-old male C57BL/6J mice were treated with UHMWPE particles and received a daily peritoneal injection of BOK-D-FMK, respectively only buffer at a dose of 3 mg/kg of body weight for 12 days until sacrifice. Bone resorption was measured by histomorphometry, micro CT (computed tomography) and TRAP-5b serum analysis. Apoptosis was measured using caspase-3 cleaved staining. The results demonstrated that UHMWPE particles induced stronger apoptotic reactions in macrophages and osteoblasts and increased bone resorption in non specifically treated mice, whereas peritoneal application of BOC-D-FMK significantly counteracted these adverse particle-related effects. We think that in particle-induced osteolysis apoptosis is pathologically increased, and that failure to reduce the quantity of apoptotic bodies leads to an up-regulation of proinflammatory cytokines, which may be responsible for the induction of osteolysis. We showed for the first time in vivo that a reduction in apoptosis leads to a significant reduction in particle-induced osteolysis. Clinically, the apoptotic cascade could become an interesting novel therapeutic target to modulate particle-induced osteolysis. PMID- 19130235 TI - The cardioprotective effect of postconditioning is mediated by ARC through inhibiting mitochondrial apoptotic pathway. AB - OBJECTIVE: Postconditioning protects the heart against ischemia/reperfusion injury by inhibiting cardiomyocyte apoptosis. However, the molecular mechanism by which postconditioning suppresses apoptosis remains to be fully understood. Apoptosis repressor with caspase recruitment domain (ARC) has been demonstrated to possess the ability to protect cardiomyocytes from apoptosis induced by ischemia/reperfusion. It is not yet clear as to whether ARC contributes to the inhibitory effect of postconditioning against cardiomyocyte apoptosis. METHODS: The cultured cardiomyocytes from 1-day old male Sprague-Dawley rats were exposed to 3 h hypoxia followed by 3 h of reoxygenation. Cells were postconditioned by three cycles each of 5 min reoxygenation and 5 min hypoxia before 3 h of reoxygenation. RESULTS: Hypoxia/reoxygenation led to a decrease of endogenous ARC protein levels. In contrast, postconditioning could block the reduction of endogenous ARC protein levels. Interestingly, inhibition of endogenous ARC expression by ARC antisense oligodeoxynucleotides reduced the inhibitory effect of postconditioning against apoptosis. Furthermore, our data showed that postconditioning suppressed the loss of mitochondrial membrane potential, Bax activation and the release of mitochondrial cytochrome c to cytosol. However, these inhibitory effects of postconditioning disappeared upon knockdown of endogenous ARC. CONCLUSION: Our data for the first time demonstrate that ARC plays an essential role in mediating the cardioprotective effect of postconditioning against apoptosis initiated by the mitochondrial pathway. PMID- 19130236 TI - Sodium selenite induces apoptosis by ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction in human acute promyelocytic leukemia NB4 cells. AB - INTRODUCTION: In this study, we delineated the apoptotic signaling pathways activated by sodium selenite in NB4 cells. MATERIALS AND METHODS: NB4 cells were treated with 20 microM sodium selenite for different times. The activation of caspases and ER stress markers, ROS levels, mitochondrial membrane potential and cell apoptosis induced by sodium selenite were analyzed by immunoblotting analysis, DCF fluorescence and flow cytometric respectively. siRNA was used to detect the effect of GADD153 on selenite-induced cell apoptosis. CONCLUSIONS: Sodium selenite-induced reactive oxygen species generation is an early event that triggers endoplasmic reticulum stress mitochondrial apoptotic pathways in NB4 cells. PMID- 19130237 TI - Sodium butyrate sensitizes human colon adenocarcinoma COLO 205 cells to both intrinsic and TNF-alpha-dependent extrinsic apoptosis. AB - Overexpression of cFLIP protein seems to be critical in the antiapoptotic mechanism of immune escape of human COLO 205 colon adenocarcinoma cells. Actually, cFLIP appears to inhibit the death receptor ligand-mediated cell death. Application of the metabolic inhibitor sodium butyrate (NaBt), short-chain volatile fatty acid, sensitized COLO 205 cells to TNF-alpha-mediated apoptosis. Western-blot analysis revealed that the susceptibility of human COLO 205 cells to apoptogenic stimuli resulted from time-dependent reduction in cFLIP and simultaneous up-regulation of TNF-R1 protein levels. Additionally, the combined TNF-alpha and NaBt treatment caused cleavage of Bid and caspase-9 activation, as well as cytochrome c release from mitochondria. Thus, the evidence of this study indicates that NaBt facilitates the death receptor signal evoked by TNF-alpha. Moreover, NaBt alone initiated intrinsic apoptosis, that in turn was abolished by intracellular BCL-2 delivery. It confirms the involvement of mitochondria in the proapoptotic activity of NaBt. The activation of mitochondrial pathway was substantiated by up-regulated expression of BAK with concomitant reduction of antiapoptotic BCL-x(L), XIAP and survivin proteins. These findings suggest that NaBt could represent a good candidate for the new therapeutic strategy aimed to improve chemo- and immunotherapy of colon cancer. PMID- 19130238 TI - Self-loading and cell culture in one layer microfluidic devices. AB - We report on a simple method for self loading and culture of mammalian cells in microfluidic multi-chambers for high throughput screening. The device was obtained by using one layer soft lithography with polydimethylsiloxane (PDMS) and thermal bonding on a glass slide. Self loading of cell suspension could be possible after degassing of the PDMS device for 30 min. Both cell loading efficiency and cell proliferation behaviors have been analyzed with triangle chambers of different sizes, all connected to the main flow channels with small entrances. We found that the number of cells loaded into the micro-chamber increased with the side length of the triangle, showing well size dependence and that self loading at a single cell level was possible for small chambers. For large chambers, the cell area density after loading and proliferation is however quite heterogeneous. For demonstration, HeLa cell growth behavior has been followed for 11 days until the total area of the largest chambers was fully filled. PMID- 19130239 TI - Sample preparation module for bacterial lysis and isolation of DNA from human urine. AB - Silica impregnated polymer monolithic columns may provide a simple method for lysing and extracting DNA from bacteria inside of microfluidic chips. Here we use Escherichia coli as a test organism for a point of care thermoplastic microfluidic module designed to take in a urine sample, mix it with lysis buffer, and perform a hybrid chemical/mechanical lysis and solid phase extraction of nucleic acids from the sample. To demonstrate proof-of-concept, we doped human hematuric urine samples with E. coli at concentrations ranging from 10(1)-10(5) colony-forming units/mL (CFU/mL) to simulate patient samples. We then performed on-chip lysis and DNA extraction. The bacterial DNA was amplified using real-time PCR demonstrating lysis and isolation down to 10(1) CFU/mL. Results were comparable to a commercial kit at higher concentrations and performed better at recovering DNA at lower concentrations. PMID- 19130240 TI - Antibody immobilization on to polystyrene substrate--on-chip immunoassay for horse IgG based on fluorescence. AB - A simple microfluidic immunoassay card was developed based on polystyrene (PS) substrate for the detection of horse IgG, an inexpensive model analyte using fluorescence microscope. The primary antibody was captured onto the PS based on covalent bonding via a self-assembled monolayer (SAM) of thiol to pattern the surface chemistry on a gold-coated PS. The immunosensor chip layers were fabricated from sheets by CO(2) laser ablation. The functionalized PS surfaces after each step were characterized by contact angle measurement, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). After the antibody-antigen interaction as a sandwich immunoassay with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody, the intensity of fluorescence was measured on-chip to determine the concentration of the target analyte. The present immunosensor chip showed a linear response range for horse IgG between 1 microg/ml and 80 microg/ml (r = 0.971, n = 3). The detection limit was found to be 0.71 microg/ml. The developed microfluidic system can be extended for various applications including medical diagnostics, microarray detection and observing protein-protein interactions. PMID- 19130241 TI - Self-standing aligned fiber scaffold fabrication by two photon photopolymerization. AB - Development of materials and fabrication techniques lead the growth of three dimensional cell culture matrices in biomedical engineering. In this work, we present a method for fabricating self-standing fiber scaffolds by two-photon polymerization induced by a femtosecond laser. The aligned fibers are 330 microm long with a diameter of 6-9 microm. Depending on the pitch of the aligned fibers, various cell morphologies are distinguished via three-dimensional images. Furthermore, the morphologies of fibroblast cells (NIH-3T3) and epithelial cells (MDCK) on the fiber scaffolds are studied to show the effect of high curvature (3 4.5 microm radii) on cell morphology. NIH-3T3 cells that contain straight pattern of actin microfilament bundles are extended and partly wrap single fibers or tend to reside between fibers. On the other hand, MDCK cells that contain circular pattern of actin microfilament bundles cover the fiber peripheral surface exhibiting high aspect ratio elongation. These results indicate that cell morphology on fiber scaffolds is influenced by the pattern of actin microfilament bundles. PMID- 19130242 TI - Patterned delivery and expression of gene constructs into zebrafish embryos using microfabricated interfaces. AB - We demonstrate a method which uses simple microfabrication and microfluidics to produce custom, shaped electroporators for the patterned delivery of foreign molecules into developing embryos. We show how these electroporators can be used to 'draw' two-dimensional patterns of tracer molecules, DNA and mRNA into the yolk and cells of zebrafish embryos (Danio rerio) at different stages of development. We demonstrate the successful delivery of patterns of Trypan Blue (normal dye), Texas Red (fluorescent dye), GFP-expressing DNA plasmids and GFP expressing mRNA constructs into both chorionated and dechorionated embryos. Both DNA and mRNA were expressed in the desired patterns subsequent to delivery. Square pulses of 10-20 V (0.20-0.40 kV/cm), 50-100 ms width were sufficient to create transient pores and introduce compounds from the late blastula period (3 hpf) to early pharyngula period (24 hpf) embryos. Using 24 hpf dechorionated embryos, we achieved a high survival of 91.3% and 89%, and a delivery efficiency of 38% and 50% for GFP-DNA and GFP-mRNA respectively. Lastly, we demonstrate the simultaneous delivery of different compounds into the developing embryo. PMID- 19130243 TI - Single particle adsorbing transfer system. AB - Here we present a novel approach for horizontal transfer of single particles after laser microdissection. The developed technique is a single particle adsorbing system for highly selective and gentle horizontal transfer of microdissected fixed and living material. As mediated via low-pressure technology, the transfer process can be precisely controlled, thus facilitating horizontal particle transfer of any isolated material, e.g. tissue material, single cells or chromosomes, in addition to precise positioning for sample release. This collection method allows one to predefine target positions and enables material transfer without contamination to any planar microchip device. This contamination free transfer is indispensable for novel lab-on-a-chip systems performing nanoscale polymerase chain reaction analyses. Using virtual reaction chamber microdevices, small amounts of microdissected material--as little as one single cell--can be directly transmitted and immediately used for single cell analysis. PMID- 19130245 TI - Human health and the environment: in harmony or in conflict? AB - Health policy frameworks usually construe environmental protection and human health as harmonious values. Policies that protect the environment, such as pollution control and pesticide regulation, also benefit human health. In recent years, however, it has become apparent that promoting human health sometimes undermines environmental protection. Some actions, policies, or technologies that reduce human morbidity, mortality, and disease can have detrimental effects on the environment. Since human health and environmental protection are sometimes at odds, political leaders, citizens, and government officials need a way to mediate and resolve conflicts between these values. Unfortunately, few approaches to applied bioethics have the conceptual tools to do accomplish this task. Theories of health care ethics have little to say about the environment, and theories of environmental ethics don't say much about human health. In this essay, I defend an approach to ethical decision-making that gives policy-makers some tools for balancing promotion of human health and protection of the environment. PMID- 19130244 TI - Integrated microfluidic devices for combinatorial cell-based assays. AB - The development of miniaturized cell culture platforms for performing parallel cultures and combinatorial assays is important in cell biology from the single cell level to the system level. In this paper we developed an integrated microfluidic cell-culture platform, Cell-microChip (Cell-microChip), for parallel analyses of the effects of microenvironmental cues (i.e., culture scaffolds) on different mammalian cells and their cellular responses to external stimuli. As a model study, we demonstrated the ability of culturing and assaying several mammalian cells, such as NIH 3T3 fibroblast, B16 melanoma and HeLa cell lines, in a parallel way. For functional assays, first we tested drug-induced apoptotic responses from different cell lines. As a second functional assay, we performed "on-chip" transfection of a reporter gene encoding an enhanced green fluorescent protein (EGFP) followed by live-cell imaging of transcriptional activation of cyclooxygenase 2 (Cox-2) expression. Collectively, our Cell-microChip approach demonstrated the capability to carry out parallel operations and the potential to further integrate advanced functions and applications in the broader space of combinatorial chemistry and biology. PMID- 19130246 TI - The concept of negotiation in shared decision making. AB - In central definitions of shared decision-making within medical consultations we find the concept of negotiation used to describe the interaction between patient and professional in case of conflict. It has been noted that the concept of negotiation is far from clear in this context and in other contexts it is used both in terms of rational deliberation and bargaining. The articles explores whether rational deliberation or bargaining accurately describes the negotiation in shared decision-making and finds that it fails to do so on both descriptive and normative grounds. At the end some notes on further analysis is given and it is suggested that the interaction is more accurately described in terms of an internal balancing of values like patient best interest, patient autonomy and patient adherence by the professional that is accepted by the patient. PMID- 19130247 TI - Trust but verify: the interactive effects of trust and autonomy preferences on health outcomes. AB - Patients' trust in their physicians improves their health outcomes because of better compliance, more disclosure, stronger placebo effect, and more physicians' trustworthy behaviors. Patients' autonomy may also impact on health outcomes and is increasingly being emphasized in health care. However, despite the critical role of trust and autonomy, patients that naively trust their physicians may become overly dependent and lack the motivation to participate in medical care. In this article, we argue that increased trust does not necessarily imply decreased autonomy. Furthermore, patients with high levels of trust and autonomy preferences are most likely to have the best health outcomes. We propose a framework for understanding simultaneous trust and autonomy preferences and for recognizing their interactive effects on health outcomes in the dynamic medical encounter. This framework argues that policy makers and health care providers should make efforts to foster not only patients' trust but also their preferences for autonomy and thus gain the best position for achieving health-related goals. PMID- 19130249 TI - Exploring the utility of an estimation procedure to reveal drug use among arrestees: implications for service delivery. AB - One of the most persistent questions plaguing researchers and service providers is how to best estimate the extent of targeted behaviors in relevant populations. One problem of particular importance is the prevalence of drug use in justice involved populations. Data have been collected through such methods as self report and analysis of biological specimens, although both have notable limitations when used alone. As a means of drawing on the strengths of both methods, such data can be used in a confirmatory manner or, alternatively, may be summed to estimate prevalence. However, this latter approach is not without difficulty as different sources lack substantial agreement. The focus of this study is to employ a methodology that utilizes multiple data sources and adjusts for nonreporting from either source. Compared to more commonly employed techniques, the results indicate that the alternative method yields higher estimates of marijuana and cocaine use among a sample of arrestees. These findings, in turn, suggest that current behavioral health interventions and policies may be based on underestimates of drug use. PMID- 19130248 TI - Craniopharyngioma: historical notes. AB - For centuries tumors have been described which today with some confidence can be diagnosed as craniopharyngiomas. It was not until 1904 Erdheim described what he called 'hypophysial duct tumors', and which Cushing later gave the name craniopharyngioma. The first operation for this neoplasm took place in 1909. It soon became evident that the outcome of surgery was rather dismal with high mortality and morbidity rates-even after corticosteroid therapy became available around 1950. Radiotherapy was introduced and later refined as radiosurgery. This paper presents a short survey of the accumulation of knowledge of craniopharyngiomas. PMID- 19130250 TI - Character analysis in cladistics: abstraction, reification, and the search for objectivity. AB - The dangers of character reification for cladistic inference are explored. The identification and analysis of characters always involves theory-laden abstraction--there is no theory-free "view from nowhere." Given theory-ladenness, and given a real world with actual objects and processes, how can we separate robustly real biological characters from uncritically reified characters? One way to avoid reification is through the employment of objectivity criteria that give us good methods for identifying robust primary homology statements. I identify six such criteria and explore each with examples. Ultimately, it is important to minimize character reification, because poor character analysis leads to dismal cladograms, even when proper phylogenetic analysis is employed. Given the deep and systemic problems associated with character reification, it is ironic that philosophers have focused almost entirely on phylogenetic analysis and neglected character analysis. PMID- 19130251 TI - Deep-brain stimulation for depression. PMID- 19130252 TI - "Oh, that's a really hard question": Australian findings on ethical reflection in an accident and emergency ward. PMID- 19130253 TI - Psychiatric ethics consultation in the light of DSM-V. PMID- 19130254 TI - Concussions, professional sports, and conflicts of interest: why the national football league's current policies are bad for its (players') health. PMID- 19130255 TI - Iron acquisition functions expressed by the human pathogen Acinetobacter baumannii. AB - Acinetobacter baumannii is a gram-negative bacterium that causes serious infections in compromised patients. More recently, it has emerged as the causative agent of severe infections in military personnel wounded in Iraq and Afghanistan. This pathogen grows under a wide range of conditions including iron limiting conditions imposed by natural and synthetic iron chelators. Initial studies using the type strain 19606 showed that the iron proficiency of this pathogen depends on the expression of the acinetobactin-mediated iron acquisition system. More recently, we have observed that hemin but not human hemoglobin serves as an iron source when 19606 isogenic derivatives affected in acinetobactin transport and biosynthesis were cultured under iron-limiting conditions. This finding is in agreement with the observation that the genome of the strain 17978 has a gene cluster coding for putative hemin-acquisition functions, which include genes coding for putative hemin utilization functions and a TonBExbBD energy transducing system. This system restored enterobactin biosynthesis in an E. coli ExbBD deficient strain but not when introduced into a TonB mutant. PCR and Southern blot analyses showed that this hemin-utilization gene cluster is also present in the 19606 strain. Analysis of the 17978 genome also showed that this strain harbors genes required for acinetobactin synthesis and transport as well as a gene cluster that could code for additional iron acquisition functions. This hypothesis is in agreement with the fact that the inactivation of the basD acinetobactin biosynthetic gene did not affect the growth of A. baumannii 17978 cells under iron-chelated conditions. Interestingly, this second iron uptake gene cluster is flanked by perfect inverted repeats and includes transposase genes that are expressed transcriptionally. Also interesting is the observation that this additional cluster could not be detected in the type strain 19606, an observation that suggests some significant differences in the iron uptake capacity between these two A. baumannii strains. Transposome mutagenesis of the strain 19606 resulted in the isolation of a derivative unable to grow under iron-chelated conditions. Gene mapping and protein analysis together with complementation assays showed that a protein related to SecA, which is a component of the Sec protein secretion system in a wide range of bacteria, is needed at least for the production of the BauA acinetobactin outer membrane receptor. Furthermore, this derivative was unable to use hemin as an iron source under limiting conditions. Taken together, these results indicate that A. baumannii expresses siderophore-mediated and hemin acquisition functions, although different isolates differ in their iron acquisition capacity. Unexpectedly, the ability of this pathogen to acquire iron depends on the expression of a SecA protein secretion function, which has not been associated with iron acquisition in bacteria. PMID- 19130256 TI - BIOMETALS 2008 (Santiago de Compostela). Preface. PMID- 19130257 TI - Quantitative imaging of metals in tissues. AB - Metals and other trace elements play an important role in many physiological processes in all biological systems. Characterization of precise metal concentrations, their spatial distribution, and chemical speciation in individual cells and cell compartments will provide much needed information to explore the metallome in health and disease. Synchrotron-based X-ray fluorescent microscopy (SXRF) is the ideal tool to quantitatively measure trace elements with high sensitivity at high resolution. SXRF is based on the intrinsic fluorescent properties of each element and is therefore element specific. Recent advances in synchrotron technology and optimization of sample preparation have made it possible to image metals in mammalian tissue with submicron resolution. In combination with correlative methods, SXRF can now, for example, determine the amount and oxidation state of trace elements in intra-cellular compartments and identify cell-specific changes in the metal ion content during development or disease progression. PMID- 19130258 TI - Sideromycins: tools and antibiotics. AB - Sideromycins are antibiotics covalently linked to siderophores. They are actively transported into gram-positive and gram-negative bacteria. Energy-coupled transport across the outer membrane and the cytoplasmic membrane strongly increases their antibiotic efficiency; their minimal inhibitory concentration is at least 100-fold lower than that of antibiotics that enter cells by diffusion. This is particularly relevant for gram-negative bacteria because the outer membrane, which usually forms a permeability barrier, in this case actively contributes to the uptake of sideromycins. Sideromycin-resistant mutants can be used to identify siderophore transport systems since the mutations are usually in transport genes. Two sideromycins, albomycin and salmycin, are discussed here. Albomycin, a derivative of ferrichrome with a bound thioribosyl-pyrimidine moiety, inhibts seryl-t-RNA synthetase. Salmycin, a ferrioxamine derivative with a bound aminodisaccharide, presumably inhibts protein synthesis. Crystal structures of albomycin bound to the outer membrane transporter FhuA and the periplasmic binding protein FhuD have been determined. Albomycin and salmycin have been used to characterize the transport systems of Escherichia coli and Streptococcus pneumoniae and of Staphylococcus aureus, respectively. The in vivo efficacy of albomycin and salmycin has been examined in a mouse model using Yersinia enterocolitica, S. pneumoniae, and S. aureus infections. Albomycin is effective in clearing infections, whereas salmycin is too unstable to lead to a large reduction in bacterial numbers. The recovery rate of albomycin-resistant mutants is lower than that of the wild-type, which suggests a reduced fitness of the mutants. Albomycin could be a useful antibiotic provided sufficient quantities can be isolated from streptomycetes or synthesized chemically. PMID- 19130259 TI - Reduction of molybdate by sulfate-reducing bacteria. AB - Molybdate is an essential trace element required by biological systems including the anaerobic sulfate-reducing bacteria (SRB); however, detrimental consequences may occur if molybdate is present in high concentrations in the environment. While molybdate is a structural analog of sulfate and inhibits sulfate respiration of SRB, little information is available concerning the effect of molybdate on pure cultures. We followed the growth of Desulfovibrio gigas ATCC 19364, Desulfovibrio vulgaris Hildenborough, Desulfovibrio desulfuricans DSM 642, and D. desulfuricans DSM 27774 in media containing sub-lethal levels of molybdate and observed a red-brown color in the culture fluid. Spectral analysis of the culture fluid revealed absorption peaks at 467, 395 and 314 nm and this color is proposed to be a molybdate-sulfide complex. Reduction of molybdate with the formation of molybdate disulfide occurs in the periplasm D. gigas and D. desulfuricans DSM 642. From these results we suggest that the occurrence of poorly crystalline Mo-sulfides in black shale may be a result from SRB reduction and selective enrichment of Mo in paleo-seawater. PMID- 19130260 TI - Iron acquisition by Pseudomonas aeruginosa in the lungs of patients with cystic fibrosis. AB - The bacterium Pseudomonas aeruginosa is commonly isolated from the general environment and also infects the lungs of patients with cystic fibrosis (CF). Iron in mammals is not freely available to infecting pathogens although significant amounts of extracellular iron are available in the sputum that occurs in the lungs of CF patients. P. aeruginosa has a large number of systems to acquire this essential nutrient and many of these systems have been characterised in the laboratory. However, which iron acquisition systems are active in CF is not well understood. Here we review recent research that sheds light on how P. aeruginosa obtains iron in the lungs of CF patients. PMID- 19130261 TI - Microbial responses to environmental arsenic. AB - Microorganisms have evolved dynamic mechanisms for facing the toxicity of arsenic in the environment. In this sense, arsenic speciation and mobility is also affected by the microbial metabolism that participates in the biogeochemical cycle of the element. The ars operon constitutes the most ubiquitous and important scheme of arsenic tolerance in bacteria. This system mediates the extrusion of arsenite out of the cells. There are also other microbial activities that alter the chemical characteristics of arsenic: some strains are able to oxidize arsenite or reduce arsenate as part of their respiratory processes. These type of microorganisms require membrane associated proteins that transfer electrons from or to arsenic (AoxAB and ArrAB, respectively). Other enzymatic transformations, such as methylation-demethylation reactions, exchange inorganic arsenic into organic forms contributing to its complex environmental turnover. This short review highlights recent studies in ecology, biochemistry and molecular biology of these processes in bacteria, and also provides some examples of genetic engineering for enhanced arsenic accumulation based on phytochelatins or metallothionein-like proteins. PMID- 19130262 TI - Molecular and genetic characterization of the TonB2-cluster TtpC protein in pathogenic vibrios. AB - TtpC is a fourth required protein in the TonB2 energy transduction system in Vibrio anguillarum. TtpC is necessary for iron transport mediated by the TonB2 system and is highly conserved in all pathogenic vibrio species studied to date as well as several marine organisms. We show here that the TtpC proteins from selected pathogenic vibrio species can function with the TonB2 system of V. anguillarum to allow iron transport mediated by a chimeric TonB2 system where the native ExbB2, ExbD2 and TonB2 function with an episomally expressed TtpC in trans from a different species. The discovery that inter-species complementation occurs can be used to identify the functional regions of the TtpC proteins and will lead to an investigation of the mechanism of interaction between the TtpC protein and other members of the TonB2 system. PMID- 19130263 TI - Iron uptake regulation in Pseudomonas aeruginosa. AB - The Pseudomonas genus belongs to the gamma division of Proteobacteria and many species produce the characteristic yellow-green siderophore pyoverdine, and often a second siderophore, of lower affinity for iron. These bacteria are known for their ability to colonize different ecological niches and for their versatile metabolism. It is therefore not surprising that they are endowed with the capacity to take up exogenous xenosiderophores via different TonB-dependent receptors. Uptake of iron is controlled by the central regulator Fur, and via extracytoplasmic sigma factors or other types of regulators (two-component systems, AraC regulators). In this review the Fur regulon (experimentally proven and/or predicted) of P. aeruginosa will be presented. An interesting feature revealed by this analysis of Fur-regulated genes is the overlap between the iron and the sulfur regulons as well with the quorum sensing system. PMID- 19130264 TI - Temporal signaling and differential expression of Bordetella iron transport systems: the role of ferrimones and positive regulators. AB - The bacterial respiratory pathogens Bordetella pertussis and Bordetella bronchiseptica employ multiple alternative iron acquisition pathways to adapt to changes in the mammalian host environment during infection. The alcaligin, enterobactin, and heme utilization pathways are differentially expressed in response to the cognate iron source availability by a mechanism involving substrate-inducible positive regulators. As inducers, the iron sources function as chemical signals termed ferrimones. Ferrimone-sensing allows the pathogen to adapt and exploit early and late events in the infection process. PMID- 19130265 TI - Genetics and environmental regulation of Shigella iron transport systems. AB - Shigella spp. have transport systems for both ferric and ferrous iron. The iron can be taken up as free iron or complexed to a variety of carriers. All Shigella species have both the Feo and Sit systems for acquisition of ferrous iron, and all have at least one siderophore-mediated system for transport of ferric iron. Several of the transport systems, including Sit, Iuc/IutA (aerobactin synthesis and transport), Fec (ferric di-citrate uptake), and Shu (heme transport) are encoded within pathogenicity islands. The presence and the genomic locations of these islands vary considerably among the Shigella species, and even between isolates of the same species. The expression of the iron transport systems is influenced by the concentration of iron and by environmental conditions including the level of oxygen. ArcA and FNR regulate iron transport gene expression as a function of oxygen tension, with the sit and iuc promoters being highly expressed in aerobic conditions, while the feo ferrous iron transporter promoter is most active under anaerobic conditions. The effects of oxygen are also seen in infection of cultured cells by Shigella flexneri; the Sit and Iuc systems support plaque formation under aerobic conditions, whereas Feo allows plaque formation anaerobically. PMID- 19130266 TI - Interacting signals in the control of hepcidin expression. AB - The amount of iron in the plasma is determined by the regulated release of iron from most body cells, but macrophages, intestinal enterocytes and hepatocytes play a particularly important role in this process. This cellular iron efflux is modulated by the liver-derived peptide hepcidin, and this peptide is now regarded as the central regulator of body iron homeostasis. Hepcidin expression is influenced by systemic stimuli such as iron stores, the rate of erythropoiesis, inflammation, hypoxia and oxidative stress. These stimuli control hepcidin levels by acting through hepatocyte cell surface proteins including HFE, transferrin receptor 2, hemojuvelin, TMPRSS6 and the IL-6R. The surface proteins activate various cell signal transduction pathways, including the BMP-SMAD, JAK-STAT and HIF1 pathways, to alter transcription of HAMP, the gene which encodes hepcidin. It is becoming increasingly apparent that various stimuli can signal through multiple pathways to regulate hepcidin expression, and the interplay between positive and negative stimuli is critical in determining the net hepcidin level. The BMP-SMAD pathway appears to be particularly important and disruption of this pathway will abrogate the response of hepcidin to many stimuli. PMID- 19130267 TI - Molecular aspects of human cellular zinc homeostasis: redox control of zinc potentials and zinc signals. AB - Zinc(II) ions are essential for all forms of life. In humans, they have catalytic and structural functions in an estimated 3,000 zinc proteins. In addition, they interact with proteins transiently when they regulate proteins or when proteins regulate cellular zinc re-distribution. As yet, these types of zinc proteins have been explored poorly. Therefore the number of zinc/protein interactions is potentially larger than that given by the above estimate. Confronted with such a wide range of functions, which affect virtually all aspects of cellular physiology, investigators have begun to elucidate the molecular mechanisms of cellular homeostatic control of zinc, especially the functions of transporter, sensor, and trafficking proteins, such as metallothioneins, in providing the correct amounts of zinc ions for the synthesis of zinc metalloproteins. The sulfur-containing amino acid cysteine in proteins has an important role in the cellular mobility of zinc ions. Sulfur-coordination environments provide sufficiently strong interactions with zinc ions; they can undergo fast ligand exchange; and they can serve as molecular redox switches for zinc binding and release. For the cellular functions of zinc, the free zinc ion concentrations (zinc potentials, pZn = -log[Zn(2+)]) and the zinc buffering capacity are critically important parameters that need to be defined quantitatively. In the cytoplasm, free zinc ions are kept at picomolar concentrations as a minute fraction of the few hundred micromolar concentrations of total cellular zinc. However, zinc ion concentrations can fluctuate under various conditions. Zinc ions released intracellularly from the zinc/thiolate clusters of metallothioneins or secreted from specialized organelles are potent effectors of proteins and are considered zinc signals. The cellular zinc buffering capacity determines the threshold between physiological and pathophysiological actions of zinc ions. When drugs, toxins, other transition metal ions or reactive compounds compromise zinc buffering, large zinc ion fluctuations can injure cells through effects on redox biology and interactions of zinc ions with proteins that are normally not targeted. PMID- 19130269 TI - The multi-layered regulation of copper translocating P-type ATPases. AB - The copper-translocating Menkes (ATP7A, MNK protein) and Wilson (ATP7B, WND protein) P-type ATPases are pivotal for copper (Cu) homeostasis, functioning in the biosynthetic incorporation of Cu into copper-dependent enzymes of the secretory pathway, Cu detoxification via Cu efflux, and specialized roles such as systemic Cu absorption (MNK) and Cu excretion (WND). Essential to these functions is their Cu and hormone-responsive distribution between the trans-Golgi network (TGN) and exocytic vesicles located at or proximal to the apical (WND) or basolateral (MNK) cell surface. Intriguingly, MNK and WND Cu-ATPases expressed in the same tissues perform distinct yet complementary roles. While intramolecular differences may specify their distinct roles, cellular signaling components are predicted to be critical for both differences and synergy between these enzymes. This review focuses on these mechanisms, including the cell signaling pathways that influence trafficking and bi-functionality of Cu-ATPases. Phosphorylation events are hypothesized to play a central role in Cu homeostasis, promoting multi layered regulation and cross-talk between cuproenzymes and Cu-independent mechanisms. PMID- 19130268 TI - Utilization of microbial iron assimilation processes for the development of new antibiotics and inspiration for the design of new anticancer agents. AB - Pathogenic microbes rapidly develop resistance to antibiotics. To keep ahead in the "microbial war", extensive interdisciplinary research is needed. A primary cause of drug resistance is the overuse of antibiotics that can result in alteration of microbial permeability, alteration of drug target binding sites, induction of enzymes that destroy antibiotics (ie., beta-lactamase) and even induction of efflux mechanisms. A combination of chemical syntheses, microbiological and biochemical studies demonstrate that the known critical dependence of iron assimilation by microbes for growth and virulence can be exploited for the development of new approaches to antibiotic therapy. Iron recognition and active transport relies on the biosyntheses and use of microbe selective iron-chelating compounds called siderophores. Our studies, and those of others, demonstrate that siderophores and analogs can be used for iron transport mediated drug delivery ("Trojan Horse" antibiotics) and induction of iron limitation/starvation (Development of new agents to block iron assimilation). Recent extensions of the use of siderophores for the development of novel potent and selective anticancer agents are also described. PMID- 19130270 TI - Biology and life cycle of Amblyomma incisum (Acari: Ixodidae). AB - Amblyomma incisum Neumann is a major tick species in the Atlantic Forest of Brazil. Tapir is the main host for adult ticks and a high aggressiveness of nymphs to humans has been reported. In this work data on the biology and life cycle of this tick species is presented for the first time. It was shown that horse is a suitable host for A. incisum adults and rabbit for larvae and nymphs. It was also shown that A. incisum is a big tick species (mean engorged female weight of 1.96 g) with a long life cycle which lasts 262.3 days when maintained at 27 degrees C and 85% RH. These laboratory conditions were, however, inappropriate and egg hatching rate (1.2%) was very low. Nevertheless egg hatching of ticks in a forest patch increased considerably (72.2%) indicating that this A. incisum population is highly dependent on a forest-like environment. PMID- 19130271 TI - Calcium-dependent signaling mechanisms and soleus fiber remodeling under gravitational unloading. AB - The decrease in postural muscle fiber size, diminishing of their contractile properties, slow-to-fast shift in myosin heavy chain expression pattern are known to be the main consequences of gravitational unloading. The Ca(2+) role in these processes has been studied for about 20 years. Ingalls et al. [J Appl Physiol 87(1):382-390, 1999] found the resting Ca(2+) level increase in soleus fibers of hindlimb unloaded mice. Results obtained in our laboratory showed that systemic or local application of nifedipine (L-type Ca(2+) channels' blocker) prevents Ca(2+) accumulation in fibers. Thus, activation of dihydropyridine calcium channels can be supposed to promote resting Ca(2+) loading under disuse. So, calcium-dependent signaling pathways may play an important role in the development of some key events observed under unloading. Since 90th the increased activities of Ca(2+)-dependent proteases (calpains) were considered as the crucial effect of hypogravity-induced muscle atrophy, which was proved later. We observed maintenance of titin and nebulin relative content in soleus muscle under unloading combined with Ca(2+) chelators administration. Nifedipine administration was shown to considerably restrict the slow-to-fast transition of myosin heavy chains (MHC) under unloading (at the RNA level and at the protein level as well). To clarify the role of calcineurin/NFAT signaling system in MHC pattern transition under unloading, we blocked this pathway by cyclosporine A application. Hereby, we demonstrated that calcineurin/NFAT pathway possesses a stabilizing function counteracting the myosin phenotype transformation under gravitational unloading. PMID- 19130272 TI - Influence of lactic acid on the proliferation, metabolism, and differentiation of rabbit mesenchymal stem cells. AB - Lactic acid, originated from degradation of biomaterials, cell cultures, and so on, would be a toxic compound in acute states. The present study was undertaken to ascertain whether the proliferation, metabolism, and differentiation of rabbit mesenchymal stem cells (rMSCs) were affected by additional lactic acid. Furthermore, this study aimed to determine whether this influence was due to decreasing pH, increasing osmotic pressure, or chemical action of lactate ion. It was shown that the proliferation and metabolism of MSCs were inhibited by decreasing pH or increasing lactate. However, when osmolarity was adjusted to the same level as that of sodium lactate using sodium chloride, cell proliferation was little affected by osmotic pressure. We also concluded that colony-forming potential and osteogenic differentiation capacity were significantly depressed by decreasing pH or increasing lactate. As was shown, this inhibition of lactate was not only due to osmotic pressure, but also mainly due to chemical action of lactate ion. However, we observed that acidifying extracellular medium and lactate ion promoted the retention of adipogenic differentiation potential of MSCs during in vitro expansion, which suggested that growth arrest and the decrease of osteogenic differentiation potential did not affect the adipogenic conversion of MSCs. PMID- 19130273 TI - Morphological and molecular aspects of physiological vascular morphogenesis. AB - The cardiovascular system plays a crucial role in vertebrate development and homeostasis. Several genetic and epigenetic mechanisms are involved in the early development of the vascular system. During embryonal life, blood vessels first appear as the result of vasculogenesis, whereas remodeling of the primary vascular plexus occurs by angiogenesis. Many tissue-derived factors are involved in blood vessel formation and evidence is emerging that endothelial cells themselves represent a source of instructive signals to non-vascular tissue cells during organ development. This review article summarizes our knowledge concerning the principal factors involved in the regulation of vascular morphogenesis. PMID- 19130274 TI - Carbon sequestration in reclaimed manganese mine land at Gumgaon, India. AB - Carbon emission is supposed to be the strongest factor for global warming. Removing atmospheric carbon and storing it in the terrestrial biosphere is one of the cost-effective options, to compensate greenhouse gas emission. Millions of acres of abandoned mine land throughout the world, if restored and converted into vegetative land, would solve two major problems of global warming and generation of degraded wasteland. In this study, a manganese spoil dump at Gumgaon, Nagpur in India was reclaimed, using an integrated biotechnological approach (IBA). The physicochemical and microbiological status of the mine land improved after reclamation. Soil organic carbon (SOC) pool increased from 0.104% to 0.69% after 20 years of reclamation in 0-15 cm spoil depth. Soil organic carbon level of reclaimed site was also compared with a native forestland and agricultural land. Forest soil showed highest SOC level of 1.11% followed by reclaimed land and agriculture land of 0.70% and 0.40%, respectively. Soil profile studies of all three sites showed that SOC pool decreased from 0-15, 15-30, and 30-45 cm depths. Although reclaimed land showed less carbon than forestland, it showed better SOC accumulation rate. Reclamation of mine lands by using IBA is an effective method for mitigating CO2 emissions. PMID- 19130275 TI - Identification and quantification of cyanobacterial toxins (microcystins) in two Moroccan drinking-water reservoirs (Mansour Eddahbi, Almassira). AB - Mansour Eddahbi (MED) (30 degrees 55'N, 6 degrees 53'W) and Almassira (ALM) (31 degrees 95'N, 6 degrees 72'W) are two Moroccan lake reservoirs located at an arid and semi-arid hydrographic basin, respectively. Both are used for irrigation, recreational activities and drinking-water production. This paper deals with the characterization and quantification of microcystins (MC) from two Microcystis aeruginosa blooms occurring in those reservoirs. The toxicity of the blooms was confirmed and evaluated by both mouse and Artemia bioassays. The calculated LD50 values revealed that the MED bloom had a medium toxicity (LD50=358 mg kg(-1) body weight), whereas the ALM bloom had low toxicity (LD50=829 mg kg(-1) body weight). The 24-h LC50 values were 1.88 and 4.15 mg ml(-1) for the MED and ALM blooms, respectively, using Artemia assay. The identification and quantification of MC variants were carried out by high performance liquid chromatography (HPLC) equipped with a photodiode array detector, and HPLC coupled to mass spectrometry. The MC content, as Microcystin-LR (MC-LR) equivalents, was higher in the MED bloom (64.4 microg g(-1) dry weight) than in the ALM bloom (9.9 microg g(-1) dry weight). Five MC variants were identified in the MED cyanobacteria bloom (MC-RR, MC-YR, MC-LR, MC-FR, and MC-WR) and only one (MC-LR) in the ALM bloom. The results show that the occurrence of toxic cyanobacteria blooms in the studied reservoirs may be regarded as a health hazard; therefore, cyanotoxin monitoring in them is highly recommended. PMID- 19130276 TI - Solid-phase extraction of Mn(II) and slurry analysis of the sorbent by electrothermal atomic absorption spectrometry. AB - In this study, a challenging preconcentration/separation method based on the sorption of manganese on ethylene glycol dimethacrylatemethacrylic acid copolymer (EGDMA-MA) treated with ammonium pyrrolidine dithiocarbamate (APDC) and its slurry analysis by electrothermal atomic absorption spectrometry was described. Optimum conditions for quantitative sorption, as well as for preparing a homogeneous and stable slurry were investigated. A 100-fold enrichment factor could be reached. The analyte element in certified sea-water and bovine-liver samples were determined in the range of 95% confidence level. The proposed technique is fast, simple, and the risk of contamination is low. The limit of detection of the method for manganese in the slurry of the blank subjected to the proposed procedure was 0.07 microg L(-1) (3delta, N:10) corresponding to 0.56 microg kg(-1) slurry. PMID- 19130277 TI - Evaluation of leaching behavior of pendimethalin in sandy loam soil. AB - The mobility of pendimethalin in sandy loam soil was studied in soil columns under laboratory conditions at two application rates, 1.0 and 2.0 kg a.i. ha(-1), with simulated rainfall of 300 mm. The maximum concentration of the herbicide was found in the top 10 cm layer, though it was found distributed in soil at all the depths at both the doses. PMID- 19130278 TI - Retrospective assessment of dryland soil stability in relation to grazing and climate change. AB - Accelerated soil erosion is an aspect of dryland degradation that is affected by repeated intense drought events and land management activities such as commercial livestock grazing. A soil stability index (SSI) that detects the erosion status and susceptibility of a landscape at the pixel level, i.e., stable, erosional, or depositional pixels, was derived from the spectral properties of an archived time series (from 1972 to 1997) of Landsat satellite data of a commercial ranch in northeastern Utah. The SSI was retrospectively validated with contemporary field measures of soil organic matter and erosion status that was surveyed by US federal land management agencies. Catastrophe theory provided the conceptual framework for retrospective assessment of the impact of commercial grazing and soil water availability on the SSI. The overall SSI trend was from an eroding landscape in the early drier 1970s towards stable conditions in the wetter mid 1980s and late 1990s. The landscape catastrophically shifted towards an extreme eroding state that was coincident with the "The Great North American Drought of 1988". Periods of landscape stability and trajectories toward stability were coincident with extremely wet El Nino events. Commercial grazing had less correlation with soil stability than drought conditions. However, the landscape became more susceptible to erosion events under multiple droughts and grazing. Land managers now have nearly a year warning of El Nino and La Nina events and can adjust their management decisions according to predicted landscape erosion conditions. PMID- 19130279 TI - Should professional ethics education incorporate single-professional or interprofessional learning? AB - Since ethical issues in the contemporary delivery of health care involve doctors, nurses, technicians, and members of other health professions, the authors consider whether members of diverse health care occupations might benefit from studying ethics in a single classroom. While interprofessional courses may be better at teaching the ethics of the relationships between and among the various health professions, single-professional courses may be better at teaching the ethics of relationships between particular kinds of professionals and patients. An ethics instructor's professional discipline affects his/her credibility with the students, and the course readings may not always be relevant to the actual work of a given discipline. With these challenges in mind, the authors suggest that the boundaries of ethics education in the health professions be reconceived to accommodate the professional mission of a specific discipline as well as the interdependence and collaboration that marks high quality health care. PMID- 19130281 TI - Metabolic adaptations of oxidative muscle during spawning migration in the Atlantic salmon Salmo salar L. AB - The adaptability/plasticity of the highly oxidative red muscle in Atlantic salmon was demonstrated during spawning migration. Substrate concentrations and the enzymatic pathways of ATP production were examined in red muscle obtained from Atlantic salmon at different sites along their migratory route in the Exploits River, Newfoundland, Canada. Individuals were chronologically sampled from a seawater site, two sites upstream, and at spawning. The 20% decrease in salmon body weight during the later stages of migration was accompanied by large decreases (mg dry weight(-1)) in both glycogen (P < 0.01) and total muscle lipid (P < 0.01). In contrast, water content and protein concentration (mg dry weight( 1)) of the red muscle increased by 25 and 34%, respectively, at spawning. Enzymes of the glycolytic pathways demonstrated a significant (P < 0.001) decrease in maximal activity as migration proceeded whereas enzymes of the oxidative phosphorylation pathways, specifically the citric acid cycle enzymes, exhibited an increase (P < 0.001) in maximal activity at spawning. The antioxidant enzyme superoxide dismutase also demonstrated an increase (P < 0.001) in maximal activity during the latter stages of migration. These adaptations imply that the red epaxial muscle of Atlantic salmon has a more efficient means of oxidizing lipids, while minimizing free radical damage, during the later stages of migration and spawning, thereby potentially increasing post spawning survival. PMID- 19130280 TI - Antioxidative role of cerium against the toxicity of lead in the liver of silver crucian carp. AB - The antioxidative role of cerium was investigated in the liver of silver crucian carp injected with lead. The fish were intraperitoneally injected with 10, 20, or 30 mg/kg wet weight PbCl(2). After a 14-day period of incubation, 35 animals were injected with a solution of 1.5 mg/kg wet weight CeCl(3). After 42 days, the wet weight and the liver weight of the fish were weighed, and the oxidative stress of the fish liver was estimated by assaying lipid peroxide, superoxide dismutase, catalase, ascorbate peroxidase, glutathione peroxidase, glutathione, ascorbic acid, and reactive oxygen species (ROS). The results show that Ce(3+) could decrease ROS accumulation, relieve the inhibition of the activities of the antioxidant enzyme and the reduction of antioxidants in fish liver caused by Pb(2+), and decrease the enhancement of hepatosomatic index of fish under various Pb(2+) dosages. PMID- 19130282 TI - Functional recombinant human anti-HAV antibody expressed in milk of transgenic mice. AB - Hepatitis A virus (HAV) is a wide spread pathogenic agent and is the common cause of acute Hepatitis A worldwide. Passive immunization of HAV plays an extremely important role in post-exposure prophylaxis with clinical applications often requiring large amounts of antibody. As an alternative to the in vitro production of recombinant proteins, expression of monoclonal antibodies (mAbs) in the milk of transgenic animals is currently used being associated with low production costs and high activity. In this paper, eight founder lines of transgenic mice were generated by co-microinjection of the two cassettes encoding the heavy- and light-chains of a neutralizing anti-HAV antibody, respectively. The expressed heavy- and light-chains of the mAb were correctly assembled and modified in the mammary gland as detected by western blotting. High expression levels of the antibody were achieved during the lactation period and found to be independent of the copy numbers of integrated transgenes. The highest level was up to 32.2 mg/ml. The binding specificity and neutralizing activity of the expressed mAb were assayed by ELISA and neutralizing test, showing that it is capable to neutralize the JN strain of Hepatitis A virus efficiently. Therefore, our results suggest that a large-scale and efficient production of the anti-HAV mAb in the milk of transgenic farm animals would be feasible in the future. PMID- 19130283 TI - Evaluation of genetic variability and mutation drift equilibrium of Banni buffalo using multi locus microsatellite markers. AB - The present study was conducted to evaluate genetic diversity of Banni buffalo and its relationship/differentiation with Murrah using genotypic data on 24 heterologus bovine specific microsatellite marker loci. A total of 138 alleles were observed with a mean of 5.75 alleles/locus across two populations. The mean observed and expected heterozygosities were found to be 0.441 and 0.572 respectively in Banni buffaloes while it was 0.464 and 0.610 respectively in Murrah buffaloes. The average heterozygosity deficit was significantly positive with substantially higher values observed in Banni (22.3%) and Murrah (24%) buffalo populations. Banni buffalo population, when evaluated for mutation drift equilibrium revealed significant heterozygosity excess under IAM while no such excess was observed under SMM and TPM. The qualitative graphical test revealed a normal L-shaped distribution of allele frequencies indicating the absence of genetic bottleneck in Banni buffaloes. The mean estimates of F-statistics over all the loci were 0.376 for F(IT), 0.187 for F(ST) and 0.232 for F(IS) respectively. Analysis of molecular variance (AMOVA) revealed 18.95% of the total variation being explained by between breed differences while 14.36% of the variation explained differences between individuals within each breed. Genotype assignment test revealed distinct clustering of Banni and Murrah buffaloes. Genetic distance was estimated using three different methods, the results of which revealed considerable genetic differentiation between these two buffalo populations. The divergence time between Banni and Murrah buffaloes was estimated to be around 7286 years. The results of the present study may be helpful in decision making for conservation programs as Banni buffalo population is on decline. PMID- 19130284 TI - Peste des petits ruminants virus (PPRV) infection; its association with species, seasonal variations and geography. AB - The present investigation included a detailed description of the factors directly associated with PPRV infection in Pakistan. A total of 1,056 suspected serum samples were analyzed for the presence of antibodies to PPRV with no history of vaccination against PPR. The samples were collected from sixty two (62) suspected outbreaks from twenty five (25) major regions of the country. Samples were collected from the animals suffering from diarrhea and showing severe respiratory signs. Competitive enzyme linked immuno-sorbant assay (cELISA) was performed to detect the presence of antibodies in the serum against PPRV. Findings suggested that the overall PPR antibody sero-prevalence recorded in sheep was 54.09% as compared to 44.15% in goats. Geography, species, sex, age and season are the major factors associated with PPRV infection. Among various age groups, the animals showed the higher prevalence (67.48% and 52.28% in sheep and goats, respectively) at >2 years as compared with the other age groups. The area-wise highest sero-prevalence was in Sindh province which was 55.10%. It was highest in the plains of Punjab and Sindh province and also in hilly areas (AJK, northern areas and northern Punjab). These findings may be correlated with variations in the sheep and goats husbandry practices within different geographic regions and the topography of different areas. The study also indicated the scenario of antibodies circulation in the population and proving that PPR is now becoming endemic and is one of the major emerging TAD in Pakistan. PMID- 19130285 TI - An in vitro nutritive evaluation and rumen fermentation kinetics of Sesbania aculeate as affected by harvest time and cutting regimen. AB - The nutritive value of Sesbania aculeate harvested after 60 and 120 days of planting and subjected to two cutting regimen (15 or 30 cm length) was evaluated by determination of the crude protein (CP), crude fibre (CF), buffer soluble nitrogen (BS-N), buffer soluble non-protein nitrogen (BS-NPN) and cell wall constituents (neutral-detergent fibre; NDF, acid-detergent fibre; ADF and lignin). In vitro digestible organic matter (IVDOM), metabolizable energy (ME), microbial nitrogen (MN) and biomass (MBM) production were also estimated in the experimental plant samples after their incubation with rumen fluid for 96 h in the absence or presence of polyethylene glycol (PEG, 6000) at a ratio of 2:1 PEG:substrate. Fermentation characteristics (initial gas production; a, gas production during incubation; b, potential gas production; a+b, fractional rate of gas production; c) were assessed using an in vitro incubation technique with rumen fluid. There was a significant (P < 0.05) effect of harvest time on all studied nutritive parameters and fermentation characteristics. The early harvest plant samples (after 60 days of planting) gave significantly higher values of IVOMD, ME, CP, BS-N, BS-NPN, MN, MBM and fractional rate of gas production and lower values of CF and cell wall constituents than the late harvest. Crude protein, BS-N, BS-NBN, IVOMD and ME were negatively correlated with CF and cell wall constituents. Metabolisable energy and IVOMD were positively correlated with CP, BS-N and BS-NPN. Cutting treatments significantly affected the CP, CF, BS-N, BS-NPN, NDF, ADF, IVDOM, ME, potential gas production and b values. There was no significant (P > 0.05) effect of added PEG on IVDOM, ME, MN, MBM, fermentation characteristics and gas production over 96 h. The greatest proportion of gas production occurred between 6 and 24 h of incubation. The fractional rate of gas production from 100 mg substrate was higher (0.046 mL/h) for the plant samples harvested at early stage and cut at 30 cm length than harvested at late stage (0.018 mL/h). C values were negatively correlated with lignin concentrations. The amount of MN and MBM produced from 100 mg substrate amounted to 1.29 mg and 14.95 mg at early maturity stage and 0.68 mg and 7.89 mg at late stage, respectively. Microbial nitrogen and MBM production were negatively correlated with CF, cell wall constituents and gas production but positively correlated with CP, BS-N and BS-NPN. PMID- 19130286 TI - The rapidly expanding role of procalcitonin as a diagnostic and prognostic assay besides in UTIs. PMID- 19130287 TI - An Escherichia coli aer mutant exhibits a reduced ability to colonize the streptomycin-treated mouse large intestine. AB - The oxygen sensor Aer of Escherichia coli affects the expression level of genes that are involved in sugar acid degradation. Phenotypes of Aer mediated gene regulation, namely growth on sugar acids was tested 'in vitro' with Phenotype MicroArrays and colonization of the mouse large intestine was tested 'in vivo'. The aer mutant did not grow on the sugar acids D: -gluconate, D: -glucuronate, D: -galacturonate, as well as the sugar alcohol D: -mannitol. Since sugar acids are the predominant carbon source for E. coli in the intestinal mucosa, the ability of the aer mutant to colonize the streptomycin-treated mouse large intestine was tested. The mutant exhibited a decreased ability to colonize the intestine when compared to wild-type cells. This effect was more pronounced under competitive conditions. The colonization phenotype of the aer mutant was complemented with either of two plasmids. One of them expressed the Aer protein, whereas the other one expressed the sugar acid degradation enzymes that are encoded by edd and eda. The data support the interpretation that decreased expression of edd and eda along with the decreased ability to grow on sugar acids may contribute to the reduced capacity of the aer mutant to colonize the mouse intestine. While Aer seems to be important during the initiation phase of colonization, FlhD/FlhC appears to be of disadvantage during maintenance phase. FlhD/FlhC is the master regulator of all flagellar genes and required for Aer expression. Mutants in flhD exhibited an initial competitive disadvantage during the first 3 days of colonization, but recovered lateron. PMID- 19130288 TI - The dynamic behavior of phycobilisome movement during light state transitions in cyanobacterium Synechocystis PCC6803. AB - Light state transition is a physiological function of oxygenic organisms to balance the excitation of photosystem II (PSII) and photosystem I (PSI), hence a prerequisite of oxygen-evolving photosynthesis. For cyanobacteria, phycobilisome (PBS) movement during light state transition has long been expected, but never observed. Here the dynamic behavior of PBS movement during state transition in cyanobacterium Synechocystis PCC6803 is experimentally detected via time dependent fluorescence fluctuation. Under continuous excitation of PBSs in the intact cells, time-dependent fluorescence fluctuations resemble "damped oscillation" mode, which indicates dynamic searching of a PBS in an "overcorrection" manner for the "balance" position where PSII and PSI are excited equally. Based on the parallel model, it is suggested that the "damped oscillation" fluorescence fluctuation is originated from a collective movement of all the PBSs to find the "balance" position. Based on the continuous fluorescence fluctuation during light state transition and also variety of solar spectra, it may be deduced that light state transition of oxygen-evolution organisms is a natural behavior that occurs daily rather than an artificial phenomenon at extreme light conditions in laboratory. PMID- 19130289 TI - Phenylalanine tolerance can already reliably be assessed at the age of 2 years in patients with PKU. AB - BACKGROUND: The clinical severity of phenylalanine hydroxylase deficiency is usually defined by either pre-treatment phenylalanine (Phe) concentration or Phe tolerance at 5 years of age. So far, little is known about the course of Phe tolerance or the ability of both pre-treatment Phe and Phe tolerance at early age to predict Phe tolerance at later age. AIM: This study was conducted to investigate the course of the individual Phe tolerance and to assess the predictive value of both the pre-treatment Phe concentration and Phe tolerance at 1 and 6 months and 1, 2, 3 and 5 years for Phe tolerance at 10 years of age. METHOD: Data on blood Phe concentration, prescribed Phe intake and weight of 213 early and continuously treated Dutch PKU patients up to 10 years of age were collected. Data acquired under good metabolic control were used in the study. Tolerance was expressed in mg/day and mg/kg per day. RESULTS: Data at 1 and 6 months and at 1, 2, 3 and 5 years of 61, 58, 59, 57, 56 and 59 patients were included for comparison with the Phe tolerance at 10 years. Phe tolerances (mg/kg per day) at 2, 3 and 5 years showed a clear correlation with the tolerance at 10 years of age (r = 0.608, r = 0.725 and r = 0.661). Results for tolerance expressed as mg/day were comparable. Pre-treatment Phe concentrations did not correlate significantly with the tolerance. CONCLUSION: Pre-treatment Phe is unreliable but Phe tolerance is a reliable predictor of the tolerance at 10 years of age, starting at 2 years of age. PMID- 19130290 TI - Pathogenic cascades in lysosomal disease-Why so complex? AB - Lysosomal disease represents a large group of more than 50 clinically recognized conditions resulting from inborn errors of metabolism affecting the organelle known as the lysosome. The lysosome is an integral part of the larger endosomal/lysosomal system, and is closely allied with the ubiquitin-proteosomal and autophagosomal systems, which together comprise essential cell machinery for substrate degradation and recycling, homeostatic control, and signalling. More than two-thirds of lysosomal diseases affect the brain, with neurons appearing particularly vulnerable to lysosomal compromise and showing diverse consequences ranging from specific axonal and dendritic abnormalities to neuron death. While failure of lysosomal function characteristically leads to lysosomal storage, new studies argue that lysosomal diseases may also be appropriately viewed as 'states of deficiency' rather than simply overabundance (storage). Interference with signalling events and salvage processing normally controlled by the endosomal/lysosomal system may represent key mechanisms accounting for the inherent complexity of lysosomal disorders. Analysis of lysosomal disease pathogenesis provides a unique window through which to observe the importance of the greater lysosomal system for normal cell health. PMID- 19130291 TI - Sepiapterin reductase deficiency in a 2-year-old girl with incomplete response to treatment during short-term follow-up. AB - Sepiapterin reductase (SR) catalyses the last step in the tetrahydrobiopterin biosynthesis pathway; it converts 6-pyruvoyl-tetrahydropterin (6-PTP) to BH(4) in an NADPH-dependent reaction. SR deficiency is a very rare autosomal recessive disorder with normal phenylalanine (Phe) concentration in blood and diagnostic abnormalities are detected in CSF. We present a 16-month-old girl with SR deficiency. From the newborn period she presented with an adaptation regulatory disorder. At the age of 3 months, abnormal eye movements with dystonic signs and at 4.5 months psychomotor retardation were noticed. Since that time axial hypotonia with limb spasticity (or rather delayed reflex development), gastro oesophageal reflux and fatigue at the end of the day has been observed. Brain MRI was normal; EEG was without epileptiform discharges. Analysis of biogenic amine metabolites in CSF at the age of 16 months showed very low HVA and 5-HIAA concentrations. Analysis of CSF pterins revealed strongly elevated dihydrobiopterin (BH(2)), slightly elevated neopterin and elevated sepiapterin levels. Plasma and CSF amino acids concentrations were normal. A phenylalanine loading test showed increased Phe after 1 h, 2 h and 4 h and very high Phe/Tyr ratios. SR deficiency was confirmed in fibroblasts and a novel homozygous g.1330C>G (p.N127K) SPR mutation was identified. On L-dopa and then additionally 5-hydroxytryptophan, the girl showed slow but remarkable progress in motor and intellectual ability. Now, at the age of 3 years, she is able to sit; expressive speech is delayed (to 1 1/2 years), passive speech is well developed. Her visual motor skills, eye-hand coordination and social development correspond to the age of 2 1/2 years. PMID- 19130292 TI - Biofilm formation by Cryptococcus neoformans under distinct environmental conditions. AB - Cryptococcus neoformans is an opportunistic fungal pathogen with a propensity to infect the central nervous system of immune compromised individuals causing life threatening meningoencephalitis. Cryptococcal biofilms have been described as a protective niche against microbial predators in nature and shown to enhance resistance against antifungal agents and specific mediators of host immune responses. Based on the potential importance of cryptococcal biofilms to its survival in the human host and in nature, these studies were designed to investigate those factors that mediate biofilm formation by C. neoformans. We observed that C. neoformans preferentially grew as planktonic cells when cultured under specific conditions designed to mimic growth within host tissues (37 degrees C, neutral pH, and ~5% CO(2)) or phagocytes (37 degrees C, acidic pH, and ~5% CO(2)) and as biofilms when cultured under conditions such as those encountered in the external environment (25-37 degrees C, neutral pH, and ambient CO(2)). Altogether, our studies suggest that conditions similar to those observed in its natural habitat may be conducive to biofilm formation by C. neoformans. PMID- 19130293 TI - Oral potassium iodide for the treatment of sporotrichosis. AB - Potassium Iodide is the antimycotic of choice for the treatment of cutaneous sporotrichosis, because of its efficacy, safety and low cost. We carried out a review of published studies on the benefits and adverse reactions of using SSKI (Saturated Solution Potassium Iodide) as treatment for sporotrichosis, but could not identify any well-designed clinical trails. There is an urgent need to conduct randomized double-blind placebo-controlled trials and critically assess usefulness of SSKI by using a standardize monitoring or an effective self-report system. PMID- 19130294 TI - Molecular cloning, polymorphism and association analyses of a novel differentially expressed porcine mRNA. AB - The mRNA differential display technique was performed to investigate the differences of gene expression in the longissimus muscle tissues from Meishan and Large White pigs. One novel mRNA that was differentially expressed was identified through semi-quantitative RT-PCR and the full-length cDNA sequence was then obtained using the rapid amplification of cDNA ends (RACE) method. The nucleotide sequence of the mRNA is not homologous to any of the known porcine genes. Sequence prediction analysis revealed that this mRNA is no-coding mRNA. Polymorphism analyses revealed that there was a C-T mutation on the position of 505 bp and PCR-HhaI-RFLP analyses revealed that Chinese indigenous pig breeds and exotic pig breeds displayed obvious genotype and allele frequency differences at this locus. Association analyses revealed that this polymorphic locus was significantly associated with the drip loss rate, water holding capacity, dressing percentage, rib numbers, lean meat percentage, estimated lean meat percentage, loin eye width and loin eye area (P < 0.05). PMID- 19130295 TI - PVAS3, a class-II ubiquitous asparagine synthetase from the common bean (Phaseolus vulgaris). AB - A gene encoding a putative asparagine synthetase (AS; EC 6.3.5.4) has been isolated from common bean (Phaseolus vulgaris). A 2.4 kb cDNA clone of this gene (PVAS3) encodes a protein of 570 amino acids with a predicted molecular mass of 64,678 Da, an isoelectric point of 6.45, and a net charge of -5.9 at pH 7.0. The PVAS3 protein sequence conserves all the amino acid residues that are essential for glutamine-dependent AS, and PVAS3 complemented an E. coli asparagine auxotroph, that demonstrates that it encodes a glutamine-dependent AS. PVAS3 displayed significant similarity to other AS. It showed the highest similarity to soybean SAS3 (92.9% identity), rice AS (73.7% identity), Arabidopsis ASN2 (73.2%) and sunflower HAS2 (72.9%). A phylogenetic analysis revealed that PVAS3 belongs to class-II asparagine synthetases. Expression analysis by real-time RT-PCR revealed that PVAS3 is expressed ubiquitously and is not repressed by light. PMID- 19130296 TI - Association study of a single nucleotide polymorphism in the exon 2 region of toll-like receptor 9 (TLR9) gene with susceptibility to systemic lupus erythematosus among Chinese. AB - Toll-like receptor 9 (TLR9) plays an important role in the induction and regulation of the innate immune system or adaptive immune responses. Genetic variations within human TLR9 have been reported to be associated with a range of immune-related diseases, such as asthma, systemic lupus erythematosus (SLE) and so on. Family-based association analysis was performed to further investigate whether a single nucleotide polymorphism (rs352140) in the exon 2 region of TLR9 gene is associated with susceptibility to SLE in a Chinese population. A total of 77 patients with SLE from 74 nuclear families, aged from 12 to 63 years, were enrolled according to 1997 criteria of American College of Rheumatology (ACR), 211 family members of these patients were also included. Genotyping was performed by PCR-restriction fragment length polymorphism (PCR-RFLP) assay. Among 77 patients with SLE, the CC, CT and TT genetype frequencies of the SNP (rs352140) were 20.8, 61.0 and 18.2%, respectively. Single loci analysis suggested that the T allele at position of rs352140 was significantly associated with the susceptibility to SLE (Z = 2.357, P = 0.018402) in dominant model, but not in additive or recessive model. Genetype analysis showed that individuals with CT genetype had greater susceptibility to SLE than those without (Z = 2.004, P = 0.045067). Our study suggests that a single nucleotide polymorphism (rs352140) in the exon 2 region of TLR9 gene may be a susceptibility factor for SLE in Chinese population. PMID- 19130297 TI - Ethical aspects of HIV/AIDS prevention strategies and control in Malawi. AB - HIV/AIDS prevention campaigns have been overshadowed by conflicting, competing, and contradictory views between those who support condom use as a last resort and those who are against it for fear of promoting sexual immorality. We argue that abstinence and faithfulness to one partner are the best available moral solutions to the HIV/AIDS pandemic. Of course, deontologists may argue that condom use might appear useful and effective in controlling HIV/AIDS; however, not everything that is useful is always good. In principle, all schools of thought and faith seem to agree on the question of faithfulness for married couples and abstinence for those who are not married. But they differ on condom use. On the ground, the situation is far more complex. We simply lack a single, entirely reliable way to resolve all disagreements regarding HIV/AIDS prevention strategies. PMID- 19130299 TI - The Center for Eukaryotic Structural Genomics. AB - The Center for Eukaryotic Structural Genomics (CESG) is a "specialized" or "technology development" center supported by the Protein Structure Initiative (PSI). CESG's mission is to develop improved methods for the high-throughput solution of structures from eukaryotic proteins, with a very strong weighting toward human proteins of biomedical relevance. During the first three years of PSI-2, CESG selected targets representing 601 proteins from Homo sapiens, 33 from mouse, 10 from rat, 139 from Galdieria sulphuraria, 35 from Arabidopsis thaliana, 96 from Cyanidioschyzon merolae, 80 from Plasmodium falciparum, 24 from yeast, and about 25 from other eukaryotes. Notably, 30% of all structures of human proteins solved by the PSI Centers were determined at CESG. Whereas eukaryotic proteins generally are considered to be much more challenging targets than prokaryotic proteins, the technology now in place at CESG yields success rates that are comparable to those of the large production centers that work primarily on prokaryotic proteins. We describe here the technological innovations that underlie CESG's platforms for bioinformatics and laboratory information management, target selection, protein production, and structure determination by X-ray crystallography or NMR spectroscopy. PMID- 19130300 TI - Sarcomas associated with hereditary nonpolyposis colorectal cancer: broad anatomical and morphological spectrum. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is primarily linked to colorectal and endometrial cancer, but is associated with a broad tumor spectrum. Though not formally part of the syndrome, occasional sarcomas have been reported in individuals with HNPCC. We used the national Danish HNPCC-register to identify HNPCC families in which sarcomas had been diagnosed. Fourteen sarcomas were identified in families with mutations in MSH2, MSH6, and MLH1. The median age at sarcoma diagnosis was 43 (15-74) years. Soft tissue sarcomas predominated followed by uterine sarcomas and eight histopathological subtypes were represented with recurrent diagnoses of liposarcoma, leiomyosarcoma, and carcinosarcoma. Tumor tissue from eight cases was available for analysis of mismatch-repair (MMR) status using immunohistochemical staining and analysis of microsatellite instability, which revealed MMR defects in six of the eight tumors investigated. This suggests that sarcomas may be part of the HNPCC tumor spectrum and that colorectal cancer should be considered in the family history of sarcoma patients. PMID- 19130301 TI - Spatial-area selective retrieval of multiple object-place associations in a hierarchical cognitive map formed by theta phase coding. AB - The human cognitive map is known to be hierarchically organized consisting of a set of perceptually clustered landmarks. Patient studies have demonstrated that these cognitive maps are maintained by the hippocampus, while the neural dynamics are still poorly understood. The authors have shown that the neural dynamic "theta phase precession" observed in the rodent hippocampus may be capable of forming hierarchical cognitive maps in humans. In the model, a visual input sequence consisting of object and scene features in the central and peripheral visual fields, respectively, results in the formation of a hierarchical cognitive map for object-place associations. Surprisingly, it is possible for such a complex memory structure to be formed in a few seconds. In this paper, we evaluate the memory retrieval of object-place associations in the hierarchical network formed by theta phase precession. The results show that multiple object place associations can be retrieved with the initial cue of a scene input. Importantly, according to the wide-to-narrow unidirectional connections among scene units, the spatial area for object-place retrieval can be controlled by the spatial area of the initial cue input. These results indicate that the hierarchical cognitive maps have computational advantages on a spatial-area selective retrieval of multiple object-place associations. Theta phase precession dynamics is suggested as a fundamental neural mechanism of the human cognitive map. PMID- 19130298 TI - Chemotherapeutic impact on pain and global health-related quality of life in hormone-refractory prostate cancer: Dynamically Modified Outcomes (DYNAMO) analysis of a randomized controlled trial. AB - PURPOSE: This paper applies the Dynamically Modified Outcomes (DYNAMO) model to a clinical trial of two chemotherapeutic regimens on global health-related quality of life (GHRQL) in hormone-refractory prostate cancer. METHODS: DYNAMO identifies the causal influences operating in a clinical trial and their mediation, moderation, and modulation by uncontrolled variables. The Southwest Oncology Group trial S9916 randomized assignment to mitoxantrone plus prednisone (M + P) versus docetaxel plus estramustine (D + E) treatments. In this application, we examine baseline-adjusted impacts of worst pain (McGill Pain Questionnaire) on GHRQL (EORTC Quality of Life Questionnaire-C30) at 10 weeks. RESULTS: The average treatment levels of pain did not differ, hence, the average mediated effect of treatment on GHRQL was zero. Nonetheless, M + P reduced the impact (the relational outcome) of pain on GHRQL by 54% relative to D + E. Individual variation in the relational outcome (modulation) was of the same magnitude as the average difference between the groups. Performance status moderated the direct effects of treatment, with D + E being more effective in good, but not poor, performance strata. CONCLUSIONS: The DYNAMO approach comprehensively accounted for treatment effects. Rather than a single average effect, there were three distinct treatment effects: one direct effect for each performance status level and a direct effect on the relationship between pain and GHRQL. PMID- 19130302 TI - Differences in heat shock protein 70 expression during larval and early spat development in the Eastern oyster, Crassostrea virginica (Gmelin, 1791). AB - For a variety of species, changes in the expression of heat shock proteins (HSP) have been linked to key developmental changes, i.e., gametogenesis, embryogenesis, and metamorphosis. Many marine invertebrates are known to have a biphasic life cycle where pelagic larvae go through settlement and metamorphosis as they transition to the benthic life stage. A series of experiments were run to examine the expression of heat shock protein 70 (HSP 70) during larval and early spat (initial benthic phase) development in the Eastern oyster, Crassostrea virginica. In addition, the impact of thermal stress on HSP 70 expression during these early stages was studied. C. virginica larvae and spat expressed three HSP 70 isoforms, two constitutive, HSC 77 and HSC 72, and one inducible, HSP 69. We found differences in the expression of both the constitutive and inducible forms of HSP 70 among larval and early juvenile stages and in response to thermal stress. Low expression of HSP 69 during early larval and spat development may be associated with the susceptibility of these stages to environmental stress. Although developmental regulation of HSP 70 expression has been widely recognized, changes in its expression during settlement and metamorphosis of marine invertebrates are still unknown. The results of the current study demonstrated a reduction of HSP 70 expression during settlement and metamorphosis in the Eastern oyster, C. virginica. PMID- 19130303 TI - Beclin-1 expression is a predictor of clinical outcome in patients with esophageal squamous cell carcinoma and correlated to hypoxia-inducible factor (HIF)-1alpha expression. AB - In the present study, we examined the relationship between Beclin-1 expression and HIF-1alpha expression in esophageal squamous cell carcinoma(ESCC). There was a loss of Beclin-1 protein expression in 33% of ESCCs. Beclin-1 expression significantly correlated with depth of invasion, lymph node metastasis and clinical stage. Among the 54 patients, The survival rate of the Beclin-1-positive group was better than that of the Beclin-1-negative group. Twenty-five of the 54 (46%) tumor specimens showed high levels of HIF-1alpha immunoreactivity. Beclin-1 expression was associated with HIF-1alpha expression. The survival rate of patients with Beclin-1-positive and HIF-1alpha-low tumors was significantly higher than that of the other groups. These results suggest that Beclin-1 and HIF 1alpha expression are important determinants of survival in ESCCs. PMID- 19130304 TI - Integrin expression levels correlate with invasion, metastasis and prognosis of oral squamous cell carcinoma. AB - The present study evaluated the relationship between alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front of oral squamous cell carcinoma (OSCC) and survival rates, as well as the clinical and pathological characteristics. Sections of 100 specimens of primary OSCC were immunostained to assess alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front. OSCC patients with higher expression levels of alpha 3, alpha 6A, and beta 1 integrin had significantly better prognosis than those with lower expression levels (median survival at low vs. high expression levels: alpha 3, 37.1 months vs. 55.7 months; alpha 6A , 38.3 months vs. 47.9 months; and beta 1, 26.1 months vs. 46.1 months) (P < 0.05). In addition, beta 1 integrin expression showed the highest correlation with clinical and pathological characteristics. This study concludes that alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front are related to the mode of invasion and prognosis in OSCC. PMID- 19130305 TI - The NAD World: a new systemic regulatory network for metabolism and aging--Sirt1, systemic NAD biosynthesis, and their importance. AB - For the past several years, it has been demonstrated that the NAD-dependent protein deacetylase Sirt1 and nicotinamide phosphoribosyltransferase (Nampt) mediated systemic NAD biosynthesis together play a critical role in the regulation of metabolism and possibly aging in mammals. Based on our recent studies on these two critical components, we have developed a hypothesis of a novel systemic regulatory network, named "NAD World", for mammalian aging. Conceptually, in the NAD World, systemic NAD biosynthesis mediated by intra- and extracellular Nampt functions as a driver that keeps up the pace of metabolism in multiple tissues/organs, and the NAD-dependent deacetylase Sirt1 serves as a universal mediator that executes metabolic effects in a tissue-dependent manner in response to changes in systemic NAD biosynthesis. This new concept of the NAD World provides important insights into a systemic regulatory mechanism that fundamentally connects metabolism and aging and also conveys the ideas of functional hierarchy and frailty for the regulation of metabolic robustness and aging in mammals. PMID- 19130306 TI - A comparison between shaker and bioreactor performance based on the kinetic parameters of xanthan gum production. AB - Xanthan gum production was studied using sugarcane broth as the raw material and batch fermentation by Xanthomonas campestris pv. campestris NRRL B-1459. The purpose of this study was to optimize the variables of sucrose, yeast extract, and ammonium nitrate concentrations and to determine the kinetic parameters of this bioreaction under optimized conditions. The effects of yeast extract and ammonium nitrate concentrations for a given sucrose concentration (12.1-37.8 g L( 1)) were evaluated by central composite design to maximize the conversion efficiency. In a bioreactor, the maximum conversion efficiency was achieved using 27.0 g L(-1) sucrose, 2.7 g L(-1) yeast extract, and 0.9 g L(-1) NH(4)NO(3). This point was assayed in a shaker and in a bioreactor to compare bioreaction parameters. These parameters were estimated by the unstructured kinetic model of Weiss and Ollis (Biotechnol Bioeng 22:859-873, 1980) to determinate the yields (Y (P/S)), the maximum growth specific rate (mu (max)), and the saturation cellular concentration (X*). The parameters of the model (mu (max), X*, m, lambda, alpha, and beta) were obtained by nonlinear regression. For production of xanthan gum in a shaker, the values of mu (max) and Y (P/S) obtained were 0.119 h(-1) and 0.34 g g(-1), respectively, while in a bioreactor, they were 0.411 h(-1) and 0.63 g g( 1), respectively. PMID- 19130307 TI - Raman spectroscopy for intracellular monitoring of carotenoid in Blakeslea trispora. AB - In the present study, we explore the feasibility of Raman spectroscopy for intracellular monitoring of carotenoid in filamentous fungi Blakeslea trispora. Although carotenoid production from this fungus has been extensively studied through various chromatographic methods and ultraviolet-visible spectroscopy, no intracellular monitoring has been demonstrated until now. The intensity of the Raman spectrum, and more conveniently that of the strongest nu(1) carotenoid band at approximately 1,519 cm(-1), exhibits a good linear correlation with the carotenoid content of the sample as determined by high-performance liquid chromatography (HPLC) and ultraviolet-visible (UV-Vis) spectroscopy. Our results suggest that Raman spectroscopy can serve as an alternative method for the study and quantification of carotenoid in batch-mated submerged cultivations of B. trispora and similar organisms. Although not as accurate as HPLC, it allows a rapid sampling and analysis, avoiding the prolonged and tedious classical isolation procedures required for carotenoid determination by HPLC and UV-Vis spectroscopy. PMID- 19130310 TI - The prognostic value of the EEG in postanoxic coma. AB - INTRODUCTION: Several studies have been performed to assess the prognostic value of early neurological and neurophysiological findings in patients with postanoxic coma, but they have not led to precise, generally accepted, prognostic rules. This study was performed to assess whether it is possible to create a prognostic outcome table, using a combination of clinical variables and the electroencephalogram (EEG). METHODS: Clinical variables and EEG were registered on admission, at day 1-5, day 6-14, and day 15-42, and were related to the Glasgow outcome scale (GOS) at 30 and 180 days. The EEG abnormalities were categorized using the EEG classification system proposed by Synek (J Clin Neurophysiol 5:161-174, 1988) and Young et al. (Can J Neurol Sci 24:320-325, 1997). These EEG classifications were then further divided into prognostic categories. RESULTS: Age was a significant predictor of outcome. The early recorded clinical variables were the most predictive and the GCS showed a limited prognostic value. The first EEG registration proved to be the most predictive. The Synek-classification was divided into three prognostic categories: "benign," "malignant," and "fatal" and the Young-classification into four: "benign," "intermediate," "malignant," and "fatal." An outcome prediction table is proposed using the Young-classification stratified for age. CONCLUSION: Age is an important variable determining the prognostic value of the EEG and should always be taken into consideration. The prognostic categories, especially when derived from the Young-classification, showed a good prognostic value. Although this is a pilot study, we believe that the revised prognostic categories have a good prognostic value in predicting outcome and are worth further investigation and validation. PMID- 19130308 TI - Nitric oxide regulates activity-dependent neuroprotective protein (ADNP) in the dentate gyrus of the rodent model of kainic acid-induced seizure. AB - The dentate gyrus (DG) of the normal rat brain contains activity-dependent neuroprotective protein (ADNP) which is widely distributed in the cytoplasm of neurons and astrocytes. Treatment with nitric oxide (NO) synthase (NOS) inhibitor N(G)-nitro-L: -arginine methyl ester (L: -NAME) caused a decrease in ADNP expression in granule cells which persisted 3 days post-treatment. However, treatment with neuronal-specific NOS inhibitor, 7-nitroindazole (7-NI), or soluble guanylyl cyclase inhibitor, ODQ, did not change ADNP expression in the DG. We have previously shown that kainic acid (KA)-induced seizure increases neuronal NOS in neurons and inducible NOS in glia cells and suppresses ADNP in the hippocampus (Cosgrave et al., Neurobiol Dis 30(3):281-292, 2008). In the DG, L: -NAME treatment prior to KA causes ADNP synthesis in granule cells by 3 h which was later restricted to the subgranular zone by 3 days. 7-NI and ODQ had no effect. Double immunostaining for neuronal marker NeuN and ADNP revealed a significant decrease of both ADNP(+) neurons and of total neuron numbers (NeuN(+)) in the hilus of animals having KA-induced seizure that had been pretreated with L: -NAME implying that NO and ADNP may act together to protect hilar neurons. Overall, these observations suggest that NO regulates ADNP in the DG under both basal and pathophysiological conditions. PMID- 19130311 TI - Safety and feasibility of percutaneous tracheostomy performed by neurointensivists. AB - INTRODUCTION: We evaluated the effects of a change from routine surgical tracheostomy (ST), performed primarily by ENT surgeons, to bedside percutaneous tracheostomy (PT) performed by neurointensivists. METHODS: The first 67 PT procedures performed by neurointensivists were retrospectively reviewed, and compared with 68 consecutive ST procedures performed during the previous year. Demographics, severity of illness, procedural complications, incidence of ventilator-associated pneumonia (VAP), duration of mechanical ventilation (DMV), length of stay (LOS), and hospital charges were evaluated. RESULTS: Age, race, gender, neurological diagnoses, comorbid illnesses, and Glasgow coma scale on admission and the day of tracheostomy were similar. Procedural complications occurred in 8% of PT patients and 9% of ST patients, including clinically significant bleeding, transient loss of the airway, ICP rise requiring treatment, or acute lung injury (P = 0.3). PT was performed earlier than ST (median [interquartile range] ventilator day 8 [4-11] vs. 12 [8-18], P = 0.001). Median DMV was shorter in the PT cohort (19 [10-27] vs. 24 [16-33] days, P = 0.02), as was median ICU LOS (15 [9-21] vs. 19 [12-27] days, P = 0.01). ICU charges (US dollars) were lower in the PT cohort (median $123,404 vs. $156,311, P = 0.01). Trends toward less VAP, shorter hospital LOS, and lower total hospital charges among patients receiving PT did not achieve significance. CONCLUSIONS: PT performed by neurointensivists was safe compared to ST. Timely PT by neurointensivists may offer significant advantages in terms of ventilator weaning, ICU LOS, and the cost of care. PMID- 19130312 TI - Huge gastric carcinoma showing an exophytic growth pattern: a case report and review of the literature. AB - CASE REPORT: We herein report a case of huge gastric carcinoma showing an exophytic growth pattern. The gastric carcinoma measured 160 x 130 mm in size. A radical resection was judged to be impossible preoperatively since the tumor invasion of the pancreas and liver was demonstrated on computed tomography. A pancreaticoduodenectomy combined with a resection of the transverse colon was performed. A pathological examination demonstrated the tumor to directly invade the pancreas and transverse colon; however, no metastasis was observed in the regional lymph nodes. The patient is alive and doing well without any recurrence at 5 years postoperatively. DISCUSSION: To obtain a better prognosis for huge gastric carcinoma showing an exophytic growth pattern, extended radical surgery is recommended since the size of the exophytic mass sometimes does not indicate the extent of the tumor. PMID- 19130314 TI - Targeting the hippocampal mossy fiber synapse for the treatment of psychiatric disorders. AB - It is widely known that new neurons are continuously generated in the dentate gyrus of the hippocampus in the adult mammalian brain. This neurogenesis has been implicated in depression and antidepressant treatments. Recent evidence also suggests that the dentate gyrus is involved in the neuropathology and pathophysiology of schizophrenia and other related psychiatric disorders. Especially, abnormal neuronal development in the dentate gyrus may be a plausible risk factor for the diseases. The synapse made by the mossy fiber, the output fiber of the dentate gyrus, plays a critical role in regulating neuronal activity in its target CA3 area. The mossy fiber synapse is characterized by remarkable activity-dependent short-term synaptic plasticity that is established during the postnatal development and is supposed to be central to the functional role of the mossy fiber. Any defects, including developmental abnormalities, in the dentate gyrus and drugs acting on the dentate gyrus can modulate the mossy fiber-CA3 synaptic transmission, which may eventually affect hippocampal functions. In this paper, I review recent evidence for involvement of the dentate gyrus and mossy fiber synapse in psychiatric disorders and discuss potential importance of drugs targeting the mossy fiber synapse either directly or indirectly in the therapeutic treatments of psychiatric disorders. PMID- 19130316 TI - Community-based study of the association of subclinical thyroid dysfunction with blood pressure. AB - The relationship between subclinical thyroid dysfunction and blood pressure has been controversial and received unsufficient attention. Thus, we performed a cross-sectional study conducted among 6,992 inhabitants from six districts of Jiangsu Province to investigate the association of subclinical thyroid dysfunction with blood pressure in China. The data from 6,583 subjects (4,115 women and 2,468 men) were included and divided into three groups: euthyroidism (n = 5669, 86.11%), subclinical hyperthyroidism (n = 108, 1.65%), and subclinical hypothyroidism (n = 806, 12.24%). In the groups with subclinical hypothyroidism and hyperthyroidism, systolic blood pressure (SBP), diastolic blood pressure (DBP), and pulse pressure were not significantly different from those in the groups with euthyroidism after being adjusted for age, sex, BMI, and smoking status (P > 0.05). More extensively, the SBP and DBP in the group of subclinical hypothyroidism with lower level of TSH (TSH 4.51-10.00 mIU/l, SCH(1)) were significantly higher than those of participants with euthyroidism (P < 0.05). Multivariable logistic analysis revealed that subclinical hypothyroidism with lower TSH (TSH 4.51-10.00 mIU/l) was an independent risk factor for increased SBP (OR = 1.28, 95% CI 1.03-1.59, P = 0.028). Similar results could not be found between groups of euthyroid and subclinical hypothyroid with higher level of TSH (TSH > 10 mIU/l, SCH(2)). Further subdivision of the euthyroid group on the basis of a TSH cut-off of 2.5 mIU/l, revealed still no significant difference in blood pressure after adjustment regardless of whether the TSH levels were in the lower reference (TSH 0.40-2.50 mIU/l, n = 4093) or in the upper reference ranges (TSH 2.51-4.50 mIU/l, n = 1576) (P > 0.05). We concluded that subclinical thyroid dysfunction was not associated with blood pressure. Neither subclinical hyperthyroidism nor subclinical hypothyroidism independently predicted increased blood pressure. PMID- 19130315 TI - Modulatory role of locus coeruleus and estradiol on the stress response of female rats. AB - The activity of the hypothalamic-pituitary-adrenal axis is modulated by the norepinephrinergic system and, in females, also by the ovarian hormones. We investigated the role of ovarian steroids and the locus coeruleus (LC) on stress induced corticosterone secretion in female rats. Ovariectomized rats without hormonal replacement (OVX) or treated with estradiol (OVE) or estradiol plus progesterone (OVEP) were subjected to jugular cannulation. Immediately after that, each hormonal treatment group was subjected to LC lesion or sham surgery or no brain surgery. After 24 h, blood samples of all 9 groups were collected before and after ether inhalation. Other four groups (OVX control, sham and lesioned, and OVE) were perfused for glucocorticoid receptor (GR) immunocytochemistry in hippocampal CA1 neurons and paraventricular nucleus (PVN). Estradiol replacement decreased while LC lesions increased stress-induced corticosterone secretion. The effect of LC lesion was potentiated with the removal of ovarian steroids. Since GR expression of lesioned animals decreased in the hippocampus, but not in PVN, we suggest that the effect of LC lesion on corticosterone secretion could be due to a reduction in the efficiency of the negative feedback system in the CA1 neurons. However, this mechanism is not involved in the estradiol modulation on corticosteroid secretion, as no change in GR expression was observed in estradiol treated animals. PMID- 19130319 TI - Retraction. Cancer stem cell hierarchy. PMID- 19130317 TI - Oligonucleotide microarray technology and its application to Mycobacterium avium subsp. paratuberculosis research: a review. AB - Microarrays represent a modern powerful technology, which have potential applications in many areas of biological research and provide new insights into the genomics and transcriptomics of living systems. The aim of this review is to describe the application of microarray technology for Mycobacterium avium subsp. paratuberculosis (MAP) research. The main focus points include a summary of results obtained for MAP using microarrays, examination of the fields of MAP research which are currently being investigated and possible areas of future research. This article is divided into two parts according to the type of nucleic acid used for array hybridisation. Articles related to MAP research using microarray technology are then divided according to the field of study, such as comparative genome analysis, diagnostics, expression or environmental studies. PMID- 19130318 TI - Structural variations in protein superfamilies: actin and tubulin. AB - Structures of homologous proteins are usually conserved during evolution, as are critical active site residues. This is the case for actin and tubulin, the two most important cytoskeleton proteins in eukaryotes. Actins and their related proteins (Arps) constitute a large superfamily whereas the tubulin family has fewer members. Unaligned sequences of these two protein families were analysed by searching for short groups of family-specific amino acid residues, that we call motifs, and by counting the number of residues from one motif to the next. For each sequence, the set of motif-to-motif residue counts forms a subfamily specific pattern (landmark pattern) allowing actin and tubulin superfamily members to be identified and sorted into subfamilies. The differences between patterns of individual subfamilies are due to inserts and deletions (indels). Inserts appear to have arisen at an early stage in eukaryote evolution as suggested by the small but consistent kingdom-dependent differences found within many Arp subfamilies and in gamma-tubulins. Inserts tend to be in surface loops where they can influence subfamily-specific function without disturbing the core structure of the protein. The relatively few indels found for tubulins have similar positions to established results, whereas we find many previously unreported indel positions and lengths for the metazoan Arps. PMID- 19130320 TI - Correlation of F-18 fluorodeoxyglucose-positron emission tomography maximal standardized uptake value and EGFR mutations in advanced lung adenocarcinoma. AB - OBJECTIVE: Epidermal growth factor receptor (EGFR) mutations in lung adenocarcinoma are involved in the tumorigenesis and regulation of cell metabolism via Akt signaling. F-18 fluorodeoxyglucose-positron emission tomography ([(18)F]FDG PET), a functional imaging modality, can be used to measure tumor cell metabolism. Thus, in this study, we hypothesize that there exist correlations between EGFR mutation status and [(18)F]FDG uptake of advanced lung adenocarcinoma. METHODS: From May 2004 to April 2008, patients with stage IIIB or IV lung adenocarcinoma who underwent [(18)F]FDG PET and EGFR mutation analysis before receiving any treatment were eligible to participate in this study. The association of EGFR mutation status with patient characteristics and the SUV(MAX) from the [(18)F]FDG PET was evaluated. Multivariate logistic regression analysis was used to analyze predictors of EGFR mutations. RESULTS: Seventy-seven lung adenocarcinoma patients were included in this study. EGFR mutations were identified in 49 (64%) of the patients. The [(18)F]FDG uptake was significantly higher in EGFR-mutant (mean SUV(MAX) = 10.5 +/- 4.7) than wild-type (8.0 +/- 3.3) lung adenocarcinoma patients (P = 0.008). The median SUV(MAX) was 9.5, and patients with an SUV(MAX) >or= 9.5 were more likely to harbor EGFR mutations (P = 0.009). In the multivariate analysis, an SUV(MAX) >or= 9.5 remained a statistically significant predictor of EGFR mutations (P = 0.005). CONCLUSIONS: Among Asian patients with advanced lung adenocarcinoma, those with higher SUV(MAX) on the [(18)F]FDG PET are more likely to carry EGFR mutations. PMID- 19130321 TI - Radiotherapy versus radiochemotherapy with cisplatin in treatment of cervical cancer. AB - AIM: To compare effects of concomitant radiochemotherapy (RCTh) and radiotherapy (RTh) alone in patients with cervical carcinoma and with 36-months follow-up analysis. MATERIAL AND METHODS: 106 stage IIA-IVA cervical carcinoma women were divided in two groups. RCTh group was treated with teleradiotherapy (50.4 Gy/T), intracavitary brachytherapy (46 Gy), and iv cisplatin (40 mg/m(2)). RTh group was treated with pelvic teletherapy (52-54 Gy/T) and intracavitary brachytherapy (50 55 Gy). RESULTS: In RCTh group absorbed radiation dose was significantly lower (50.4 vs. 52.7 Gy) and the duration of treatment was significantly shorter (45.1 days vs. 47.8 days). There were no statistical differences in both groups in survival (59% in RCTh group vs. 56% in RTh group), response to treatment (86% vs. 90%), local recurrence (42% vs. 49%) metastasis occurrence (21% vs. 17%), anemia (40% vs. 26%), early postradiation reactions in intestines (77% vs. 65%) and bladder (71% vs. 61%) as well as in incidence of rectovaginal (10% vs. 4%) and vesicovaginal formation of fistulas (6% vs. 4%), respectively. There were significant differences between two groups in: nausea (77% vs. 6%), vomiting (65% vs. 3.7%), leucopenia (69% vs. 26%) and thrombocytopenia (35% vs. 9%), and late postradiation bladder effects (94% vs. 74%). RCTh patients with anemia had lower 36-months survival rates (42% vs. 71%), more frequent local recurrences (77% vs. 31%) and metastasis-free survival rates (61% vs. 90%) than RCTh patients without anemia. CONCLUSIONS: RCTh gives better treatment results in patients without than in patients with anemia and higher overall survival rates than RTh in patients without anemia. The only clinical prognostic factor for advance cervical carcinoma is the clinical stage of the disease. PMID- 19130322 TI - Shorter survival-times following adjuvant endocrine therapy in oestrogen- and progesterone-receptor positive breast cancer overexpressing HER2 and/or with an increased expression of vascular endothelial growth factor. AB - PURPOSE: To investigate the possible correlation between expression of HER2 and vascular endothelial growth factor (VEGF), and to determine the predictive value of these factors in patients receiving adjuvant endocrine therapy including the group with a breast cancer (BC) positive for both oestrogen receptor (ER) and progesterone receptor (PgR). MATERIAL AND METHODS: By enzyme immuno-sorbent assays (ELISA) tumour levels of HER2 and VEGF proteins were determined in 679 consecutive primary BC patients, median age 63 years, median follow-up time 92 months. A total of 404 patients received adjuvant endocrine therapy, mainly tamoxifen, out of them 295 had an ER and PgR positive BC. In 160 patients, HER2 status was also determined by immunohistochemistry (IHC) using the monoclonal antibody CB11. RESULTS: Overexpression of HER2 by IHC was found in 15% of the patients. Overexpression of HER2 by ELISA correlated with HER2 by IHC (P < 0.001) and a higher VEGF expression (P = 0.004). Patients receiving adjuvant endocrine therapy with high VEGF (RFS P = 0.0087, BCCS P = 0.0012) or over-expressing HER2 (RFS P = 0.0116, BCCS P = 0.0036) had significantly shorter survival. Factors retaining statistical significance in multivariate analyses for recurrence-free survival (RFS) were nodal status (P < 0.001), tumour size (P = 0.005) and VEGF (P = 0.032) and for breast cancer corrected survival (BCCS) nodal status (P < 0.001), tumour size (P = 0.001), ER status (P = 0.022), and VEGF (P = 0.016). Both factors were significantly correlated with survival in the group with a BC positive for both ER and PgR; VEGF (RFS P = 0.0177, BCCS P = 0.0321) and HER2 (RFS P = 0.0143, BCCS P = 0.0311). In multivariate analyses, nodal status (P < 0.001) and VEGF (P = 0.021) were independent factors for RFS. Nodal status (P < 0.001) and tumour size (P = 0.016) retained independent factors for BCCS. Combined analysis identified a high-risk group (HER2 positive and high VEGF) with significantly reduced survival. CONCLUSION: The results from this retrospective analysis suggest that overexpression of HER2 and higher VEGF expression may add information on patient's outcome after adjuvant endocrine therapy in ER and PgR positive BC. PMID- 19130323 TI - An unusual case of smoldering AML with prolonged indolent clinical course and spontaneous remission in the terminal phase. AB - An unusual case of acute myeloblastic leukemia (AML) with indolent clinical course and spontaneous remission in the terminal phase is described. A 63-year old male has been diagnosed to suffer from AML, subtype M2. Chromosomal analysis showed 46,XY,del(6)(q21). Clinical course was slowly progressive ("smoldering" AML). The patient did not require cytoreductive drugs, and occasional supportive therapy was his only treatment. Five years from diagnosis patient exhibited spontaneous remission of the disease, accompanied with disappearance of del(6q) clone. Six months after, relapse occured and patient died from CNS bleeding. Additional curiosity in this case is the fact that patient's older brother died of acute lymphoblastic leukemia at the age of 71 years. Possible mechanisms of spontaneous remission of AML and genetic predisposition for human leukemia are discussed with a review of the literature. PMID- 19130324 TI - A study on the CD(4) (+) CD (25) (+) regulatory T cells in patients with gastrointestinal malignancies. AB - OBJECTIVES: Regulatory T cells play an active role in the maintenance of the immune system's tolerance of both foreign and self antigens. Particularly, CD(4) (+) CD(25) (+) regulatory T cells participate in tumor immunity. The study provided further evidence on the involvement of CD(4) (+ )CD(25) (+) regulatory T cells in immune system impairment in patients with gastrointestinal malignancies. METHODS: Using flow cytometry, CD(4) (+ )CD(25) (+) regulatory T cells were analyzed in peripheral blood from 114 patients with gastrointestinal malignancies and 15 healthy controls. RESULTS: The prevalence of the CD(25) (+) subset in CD(4) (+) T cells was increased in patients with colorectal carcinoma compared with healthy controls. The phenotic characteristics of the CD(4) (+ )CD(25) (+) T cells in patient with malignancies were low expression of CD(45) RA and no expression of CD(69). Our results indicated that when compared with healthy control, the proportions of CD(4) (+) CD(25) (+) T cells in the peripheral blood of patients with colorectal, gastric, and esophageal carcinoma were significantly higher (P < 0.05) in colorectal carcinoma (22.11 +/- 9.65%), gastric carcinoma (17.74 +/- 4.24%), and esophageal carcinoma (24.37 +/- 4.82)%, respectively. Further analysis on the proportion of CD(4) (+ )CD(25) (+) T cells revealed that those patients with gastrointestinal malignancies in stages IV were higher than those of in stage I-III, though no significant difference was observed (P > 0.05). However, the proportion of CD(4) (+ )CD(25) (+) T cells in the patients with relapse gastric carcinoma (23.32 +/- 4.98%) was significantly higher than that of patients with primary gastric carcinoma (P < 0.01). CONCLUSIONS: The increased CD(4) (+ )CD(25) (+) T cells in patients with gastrointestinal malignancies may be related to immunosuppression and tumor progression. This suggests that elimination or reduction of CD(4) (+ )CD(25) (+) regulatory T cells can improve effective tumor immunity for immunotherapy. PMID- 19130325 TI - Patient empowerment through the internet. PMID- 19130326 TI - The clinical significance of diffusion-weighted MR imaging in stroke and TIA patients. AB - BACKGROUND AND PURPOSE: Diffusion-weighted magnetic resonance imaging (DWI) is an advanced imaging technique that allows non-invasive evaluation of water diffusibility in brain tissue. The following report focuses on the clinical significance of DWI in stroke and TIA patients SUMMARY OF REVIEW: (1) TIA patients demonstrate DWI lesions at a rate of 1 in 6 to 2 in 3. Symptom duration, speech or motor symptoms and aetiology seem to correlate with the rate of DWI positivity. (2) In stroke patients, the DWI detection rate of ischaemic lesions is >95%. Small lesion size and location in the brainstem increase the risk of false-negative DW-images. A negative DW-image in a patient with stroke-like symptoms should stimulate the search for an alternative diagnosis. However, one half of such patients can be expected to have ischaemic stroke as the best final diagnosis. (3) Infarct age determination based on DWI characteristics is not possible in the first few hours. However, the combined interpretation of DWI images and apparent diffusion coefficient (ADC) maps enables the distinction of infarcts ?5 day old from infarcts >10 days old. On average in DW-images alone, the hyperintense signal disappears after two months. Normalisation can occur as early as one month and as late as four and a half months. (4) DWI lesion size is a prognostic marker of stroke outcome. However, in a mixed stroke population, outcome prediction by DWI cannot replace clinical outcome scores. (5) The mismatch concept hypothesises that DWI lesions reflect irreversibly infarcted tissue that is surrounded by an area of reduced perfusion. The larger the perfusion-diffusion mismatch the more tissue is potentially salvageable, e.g., by early reperfusion. Although this concept is appealing, more recent data have shown that DWI lesions are not necessarily irreversibly damaged tissue and that perfusion abnormalities tend to overestimate the ischaemic penumbra. More recently, the mismatch between clinical stroke severity as measured with the NIH stroke Scale Score (NIHSSS) and the volume of DWI lesions has been introduced. (6) In posterior circulation stroke, DWI lesion detection rate is significantly lower than in anterior circulation stroke. (7) DWI features provide important information about stroke aetiology. Multiple DWI lesions in more than one circulation suggest cardioembolism. However, this assignment should be restricted to DWI lesions showing the same appearance on ADC-maps. In patients with lacunar syndromes, every fourth to sixth patient can be expected to have >1 DWI lesion, indicating an embolic mechanism. Thus, DWI findings may be clinically useful to tailor the aetiological work-up, which may result in early implementation of specific treatment for secondary stroke prevention. (8) DWI may detect clinically silent ischaemic lesions after carotid interventions. A systematic review reported the rate of new DWI lesions as being significantly higher in carotid stenting patients (37%) compared to carotid endarterectomy patients (10%). As caveats, all studies included were non randomized trials. In addition, the clinical significance of these lesions is unclear. Studies, comparing the risk of silent ischaemia in carotid stenting versus endarterectomy patients and evaluating the value of DWI as surrogate marker in a randomised, prospective setting are currently under way. CONCLUSION: DWI provides clinically useful information and has the means to improve the quality of diagnosis, treatment, and outcome prediction in stroke and TIA patients. PMID- 19130327 TI - Non-physicians may reach correct diagnoses by using Google: a pilot study. AB - OBJECTIVE: We endeavoured to determine whether individuals who are not physicians are likely to arrive at correct diagnoses by using Internet resources. METHODS: In this prospective study four non-physicians used Google to search for diagnoses. They reviewed the 26 diagnostic cases presented in the case records of the New England Journal of Medicine during 2005; they were blind to the correct diagnoses. The main measurement was the percentage of correct diagnoses arrived at by non-physicians by using Google. The diagnostic success of the four non physicians was compared to that of four young physicians. RESULTS: The average diagnostic success of non-physicians was 22.1% (95% confidence interval [CI] 4.5 39.7%). There was no statistically significant difference between the non physicians regarding this outcome (p = 0.11). They took 8.9 +/- 6.7 (mean +/- standard deviation) minutes for case record reading and 17.4 +/- 7.9 minutes for Google searching per case. Non-physicians performed worse than physicians (50.9% [95% CI 37.4-64.5%]) in regard to diagnostic success (p <0.001). CONCLUSION: Non physicians, at least those who have similar characteristics to the participants in the present study, may occasionally reach correct diagnoses by performing a brief web-based search. Doctors should realise that patients may assume a more active role in their health decision-making process and take this development into consideration in physician-patient interaction. PMID- 19130328 TI - Staged surgical therapy of basal cell carcinoma of the head and neck region: an evaluation of 500 procedures. AB - QUESTIONS UNDER STUDY / PRINCIPLES: The surgical therapy of basal cell carcinoma (BCC) is especially demanding in the facial area. This retrospective study was undertaken to evaluate the outcome of staged surgical therapy (SST) of BCC of the head and neck region performed on an interdisciplinary basis at our institution. METHODS: Patients treated for BCC in the head and neck area between 1/1/1997 and 31/12/2001 were included in the study. The lesions were histologically evaluated. Diameter of lesion, number of stages, defect coverage, operation time, and recurrence and infection rates were analysed using descriptive and inferential statistical procedures. RESULTS: 281 patients were included in the study. SST was performed in two stages in 43.7%, in three stages in 12.9% and in four or more stages in 2.7%, depending on the type of tumour and the patient's pretreatment status. The total operating time per lesion averaged one hour. Defect coverage was achieved by direct closure (37.7%), by full thickness skin graft (39.5%), by split skin graft (1.1%), by local flaps (20.3%) or by composite grafts (1.1%). Median follow-up time was 58.5 months. Low rates of recurrence (3.6%) and infection (2%) were observed with this technique. CONCLUSIONS: The staged surgical therapy of basal cell carcinoma evaluated here offers a series of advantages in respect of patient comfort and safety and economy, while allowing precise histological safety with low infection rates and reliable long-term results. PMID- 19130329 TI - Uptake of breast cancer screening measures among immigrant and Swiss women in Switzerland. AB - BACKGROUND: Early detection of breast cancer plays a crucial role in survival, and in most developed countries immigrant women present for treatment at a later stage of the disease. Compared to the indigenous population, immigrant women have a lower uptake of breast cancer prevention services. The situation for immigrant women in Switzerland is compounded by the fact that all women living in Switzerland are at major risk of developing breast cancer during their lifetime. QUESTION UNDER STUDY: Our study aimed to detect disparities in uptake of preventive methods such as mammography, clinical breast examination (CBE) and breast self-examination (BSE) among women from the four major immigrant groups Italy, Spain, former Yugoslavia and Portugal, compared to the native population in Switzerland. METHODS: This study is a secondary analysis of data from the 2002 Swiss Health Survey, a nationwide cross-sectional telephone survey. The self reported questionnaires addressed immigrant and Swiss women of age 20 and over (n = 9,790). Descriptive statistics were used to describe the demographic characteristics of the sample. The dichotomous variables of interest, mammography, CBE and BSE were adjusted for nationality and further sociodemographic factors using logistic regression, with Swiss women serving as the comparison group. RESULTS: Main predictors for mammography uptake were higher income and older age. For the use of CBE, lower age, higher income and nationality were the strongest predictors. Women from former Yugoslavia (OR = 2.6) and Portugal (OR = 2.8) more frequently stated that they did not receive CBE compared to Swiss females (p = .011). BSE is linked to a higher socioprofessional status and to nationality. Women from Italy (OR = 1.62) and former Yugoslavia (OR = 2.6) perform BSE significantly less often than Swiss women (p = .0001). CONCLUSION: Differences exist in the use of mammography, CBE and BSE among the four major foreign nationality groups living in Switzerland compared to Swiss nationals. Immigrant and low-income women might draw benefit from a systematic and culturally adapted breast cancer screening programme to overcome disparities in access to screening. PMID- 19130330 TI - Effects of additional intra-abdominal organ injuries in patients with penetrating small bowel trauma on morbidity and mortality. AB - BACKGROUND: We aimed to evaluate the effects on morbidity and mortality of additional organ injuries obtained concurrently with penetrating small bowel injuries. METHODS: Between January 2000 and March 2005, patients in whom penetrating small bowel injuries occurred as a result of abdominal injuries were allocated into two groups and assessed. The first group included patients with isolated small bowel injuries, while those in the second group had small bowel injuries together with other intra-abdominal organ injuries. RESULTS: Small bowel injury was identified in 38 patients (34 M, 4 F) with an average age 38.1+/-8.86 (17-53) years (Group I: 20 patients; Group II: 18 patients). The PATI (penetrating abdominal trauma index) values of the first and second groups were 6.2+/-2.58 and 17+/-6.36, respectively, and the difference was statistically significant (p<0.001). Complication rates were 23.68% for the whole group, 5% in group I and 44.8% in group II. The rate was significantly higher in group II (p<0.01). Mortality was 2.63% and it was not related to intestinal injury. CONCLUSION: Isolated penetrating small bowel injuries are amenable to treatment with lower complication rates. Additional organ injury increases the development of complications. Injury severity score (ISS) and PATI may be useful for estimating the risk of development of complications. PMID- 19130331 TI - Tissue oxidative stress level and remote organ injury in two-hit trauma model of sequential burn injury and peritoneal sepsis are attenuated with N-acetylcysteine treatment in rats. AB - BACKGROUND: The second hit in trauma leads to an exaggerated inflammatory response and multiple organ failure. Infection following burn injury is a useful model for two-hit trauma studies. The aim of this study was to investigate the effect of N-acetylcysteine (NAC) treatment as an antioxidant in a two-hit trauma model. METHODS: 30% scalding burn injury was performed in 45 rats and cecal ligation-puncture (CLP) was performed 72 hours later. Groups were allocated as follows: Group I: No treatment was performed; Group II: Rats were treated with 150 mg/kg/day i.p. NAC for 72 h following CLP; Group III: Rats were treated with 150 mg/kg/day i.p. NAC for 6 days following thermal injury. Tissue samples were collected to study the tissue malonyldialdehyde (MDA) and glutathione (GSH) levels, and for histopathological examination on day 7. RESULTS: No difference in mortality between groups was detected. Tissue MDA levels significantly decreased in the liver (p=0.01, p=0.02) and ileum (p=0.01, p=0.02) in the treatment groups. Lung tissue GSH levels were found to be significantly increased in Group II (p=0.02). Lung injury scores were decreased in Group II (p=0.005) compared to the control group. CONCLUSION: NAC attenuated tissue oxidative stress level and remote organ injury in two-hit trauma. Further experimental and clinical studies on this subject are necessary. PMID- 19130332 TI - Is the zeolite hemostatic agent beneficial in reducing blood loss during arterial injury? AB - BACKGROUND: Uncontrolled hemorrhage is the leading cause of fatality. The aim of this study was to evaluate the effect of zeolite mineral (QuikClot - Advanced Clotting Sponge [QC-ACS]) on blood loss and physiological variables in a swine extremity arterial injury model. METHODS: Sixteen swine were used. Oblique groin incision was created and a 5 mm incision was made. The animals were allocated to: control group (n: 6): Pressure dressing was applied with manual pressure over gauze sponge; or QC group (n: 10): QC was directly applied over lacerated femoral artery. Mean arterial pressure, blood loss and physiological parameters were measured during the study period. RESULTS: Application of QC led to a slower drop in blood pressure. The control group had a significantly higher increase in lactate within 60 minutes. The mean prothrombin time in the control group was significantly increased at 60 minutes. The application of QC led to decreased total blood loss. The QC group had significantly higher hematocrit levels. QC application generated a significant heat production. There were mild edematous and vacuolar changes in nerve samples. CONCLUSION: According to the physiological parameters, we observed that zeolite tends to reduce blood loss, however could not stop bleeding completely. We believe that further clinical trials are needed to conclude that zeolite could be used in the routine practice. PMID- 19130333 TI - The association of microalbuminuria and duration of mechanical ventilation in critically ill trauma patients. AB - BACKGROUND: Microalbuminuria, which predicts organ failure, probably predicts the necessity of mechanical ventilation as well as its duration. The primary goal of this study was to determine the incidence and presence of a relationship between microalbuminuria and duration of mechanical ventilation in trauma intensive care unit (ICU) patients. METHODS: Sixty admitted critically ill trauma patients were categorized by duration of mechanical ventilation to group A (< or =7 days) or group B (>7 days) in this prospective observational study. We measured serial spot urine micro-albumin-creatinine ratios (ACR) on admission to the ICU (ACR-1), at 24, 72, and 120 hours after admission, and on the days of beginning and discontinuation of mechanical ventilation (ACR-2 and ACR-3, respectively). RESULTS: Seventy percent of the patients had microalbuminuria and 63.3% had an ACR > or =100 mg g-1 at admission. ACR-1 [mean (SE)] in patient groups A and B was 30 (6) and 63 (16) mg g(-1) (p=0.0002); ACR-2 was 40 (4) and 52 (8) (p=0.007); and ACR-3 was 30 (11) and 44 (11), respectively (p=0.023). For all patients, mean (SE) ACR-2 fell from 37 (4) to 34 (8) mg g(-1) on the day of discontinuation of mechanical ventilation (p<0.01). CONCLUSION: The ACR can be used in estimating the risk of prolonged mechanical ventilation, even on the first day of admission of critically ill trauma patients. PMID- 19130334 TI - Comparison of POSSUM and P-POSSUM for risk-adjusted audit of patients undergoing emergency laparotomy. AB - BACKGROUND: The Physiological and Operative Severity Score for Enumeration of Mortality and Morbidity (POSSUM) scoring system, derived from a heterogeneous population, has been used successfully as an audit tool, but it has appeared to over-predict mortality in low-risk groups for which an alternative system, the Portsmouth predictor equation for mortality (P-POSSUM) was designed and used successfully. In this prospective study, we compared these two equations in predicting death and tested their validity in predicting morbidity and mortality in patients undergoing emergency laparotomy in a tertiary hospital. METHODS: Eighty-two patients who underwent emergency laparotomy were included in this study. Actual morbidity and mortality rates were compared with the predicted mortality and morbidity rates using both POSSUM and P-POSSUM equations by linear regression and exponential methods of analysis. RESULTS: POSSUM equation significantly over-predicted both morbidity and mortality when linear regression analysis was used, but predicted well when exponential analysis was used. Prediction of mortality by P-POSSUM was good using both linear and exponential analyses, and exponential method was better than linear regression analysis. CONCLUSION: P-POSSUM is a better equation than POSSUM in predicting mortality, and exponential method is better than linear regression analysis. Both equations are useful tools for risk-adjusted surgical audit of patients undergoing emergency laparotomy. PMID- 19130335 TI - Accuracy of sonography in detection of renal injuries caused by blunt abdominal trauma: a prospective study. AB - BACKGROUND: This prospective study was conducted to evaluate the accuracy of sonography in detection of renal injuries caused by blunt abdominal trauma. METHODS: One hundred sixty-four patients (131 M, 33 F) with a history of recent blunt abdominal trauma who were stable enough to undergo both sonography and CT scan were included in this study. All of the cases had accepted indications for renal imaging. Ultrasound, as simultaneous gray scale B-mode scan and color Doppler study, was achieved in all of the patients as the first imaging modality. Considering CT scan as the imaging modality of choice in evaluation of renal injuries caused by trauma, sonography findings were compared with CT scan results. RESULTS: Of the 164 patients referred for kidney sonography and CT scan, renal damage was detected in 103 cases by CT scan (63%). In 14 patients (13.5%), bilateral renal injuries were identified. Considering grading classification proposed by the American Association for the Surgery of Trauma (AAST), 57%, 24%, 9.5%, 6% and 3.5% of renal injuries were diagnosed as grade I, II, III, IV and V, respectively, by CT scan. Of the 164 patients, ultrasound results were consistent with renal damages in 66 cases (40%). Of these patients, signs of parenchymal hematoma, perinephric hematoma and pelvocaliectasis associated with internal echogenicity were the most prevalent ultrasound findings. Overall sensitivity and specificity of sonography in detection of renal injures were 48% and 96%, respectively, with a 0.8 positive predictive value, a 0.57 negative predictive value and an overall accuracy of 79%. CONCLUSION: In spite of the availability and ease of performance of sonography in evaluation of trauma victims, this imaging modality has low sensitivity in detection of renal injuries and overlooks significant damages. CT scan should be considered as the diagnostic modality in victims of kidney trauma who are hemodynamically stable and have clear indications for renal imaging. PMID- 19130337 TI - Thermogenic and metabolic response to amino acid solution in brain-dead patients. AB - BACKGROUND: There is a strong correlation between amino acid infusion and increase in energy expenditure and thermogenesis. In this study, the effects of amino acid solutions on thermogenesis and energy expenditure were investigated in intensive care patients who were diagnosed as brain death. METHODS: Twenty-six brain-dead patients were included in the study. The patients received a 4 h infusion of standard amino acid solution. Energy expenditure was measured by indirect calorimetric method. ANOVA and Bonferroni tests were used for statistical analysis. RESULTS: The predicted energy expenditure (PEE) values were higher than the resting energy expenditure (REE) values in 24 patients. The mean REE/PEE value in all patients indicated that there was a 29% decrease in REE when compared to PEE. Body temperature and energy expenditure values were not increased during the parenteral infusion of amino acid solutions. There was no statistically significant difference between mean metabolic measurements and basal values at any time interval during the study period. CONCLUSION: We conclude that the stimulatory effect of amino acid solution on energy expenditure and thermogenesis is abolished in brain-dead patients. This result may indicate a possible central pathway responsible for the increase in energy expenditure and thermogenesis. PMID- 19130336 TI - Early decompressive craniectomy for neurotrauma: an institutional experience. AB - BACKGROUND: Neurotrauma centers have developed management protocols on the basis of evidence obtained from literature analysis and institutional experience. This article reviews our institutional experience in the management of severe traumatic brain injury (TBI) at Simon Bolivar Hospital, the district trauma center for Bogota's north zone. METHODS: This is a case control study comparing a group of patients (n: 16) operated for severe TBI between January 2002 and July 2004 according to an institutional management protocol characterized by an early decompressive craniectomy (DC) approach versus a historical control group (n: 20) managed before the implementation of such protocol. Mortality and Glasgow Outcome Score (GOS) at 6 months were used as the main outcome variables. RESULTS: An early DC protocol implemented within 12 hours from injury in 16 patients with severe isolated TBI and a Marshall score between III or IV was associated with a lesser mortality than the conventional approach with ventriculostomy and Intensive Care Unit (ICU) management alone. The GOS was significantly better in the DC group (p=0.0002) than in the control group. CONCLUSION: The use of an early DC protocol for severe TBI patients (Glasgow Coma Scale <9) had a significantly improved outcome compared with the conventional approach with ventriculostomy and ICU management in Simon Bolivar Hospital in Bogota, Colombia. PMID- 19130338 TI - Laparostomy in patients with severe secondary peritonitis. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness of laparostomy with the Bogota bag for the management of patients with severe secondary peritonitis and the risk factors for survival. METHODS: Thirty-seven patients (22 males, 15 females; mean age 63.5; range 44 to 83 years) with secondary peritonitis were treated by laparostomy and temporary closure with Bogota bag. APACHE II scores and Mannheim Peritonitis Index (MPI) were used to calculate the disease severity. The outcomes and effectiveness of APACHE II and MPI values were analyzed retrospectively. RESULTS: The mortality rate was 43.2%. Significant differences were noted between survivors and non-survivors according to initial APACHE II and MPI scores and the number of operations. The non survivors had higher APACHE II (r=0.81, p=0.001) and MPI (r=0.39, p=0.02) scores. The patients who survived were re-operated 1.6 times and those who died were re operated 4.7 times. In five patients, laparostomy was closed primarily, while in the others, the wound was left open to heal secondarily. CONCLUSION: Patients with higher APACHE II and MPI scores and number of operations had higher rates of mortality due to their major risk factors. Temporary abdominal closure using the Bogota bag in patients with secondary peritonitis is an inexpensive-simple method, permitting evaluation of underlying viscera and recognition of infection. PMID- 19130339 TI - Preputial skin can be used in all boys with burns requiring grafting. AB - BACKGROUND: In patients with extensive burns, all unburned areas can be used as donor sites. In male patients with small burns, preputial skin graft may be a good choice in order to avoid donor site problems. METHODS: We retrospectively reviewed 12 patients treated in our burn center from January 1997 to August 2007 with preputial skin grafting. RESULTS: In three patients, preputial skin was the only graft source. In nine patients, other donor sites were also used for split thickness graft harvesting. In two patients, preputial skin graft was used for periareolar grafting where hyperpigmented healing was preferred. In the rest of the patients, preputial skin graft was used on various parts of the body. Overall success rate for preputial skin graft was 100%. There was no early donor site morbidity. CONCLUSION: Preputial skin is a full-thickness, highly elastic and easy-to-harvest graft that should be considered in all burned male children in whom grafting is needed. PMID- 19130340 TI - [Early laparoscopic cholecystectomy for acute cholecystitis]. AB - BACKGROUND: The aim of this study was to assess the clinical results of treatment by laparoscopic cholecystectomy (LC) in patients with acute cholecystitis. METHODS: Between 1994 and 2006, LC was performed in 3876 patients in Selcuk University Meram Medicine Faculty. The clinical, biochemical, radiologic, and operative data of 182 (101 F, 81 M) consecutive patients with acute cholecystitis operated 3 days after the onset of symptoms were analyzed retrospectively to determine the complications and morbidity after operation. RESULTS: The conversion rate was 31 (17.03%) in acute cholecystitis. Postoperative length of stay was found as 4 days in the successful LC group and 7 days in the conversion group. For acute cholecystitis, we found a statistical difference between the successful LC group and the conversion group in terms of length of postoperative hospitalization time and gallbladder wall thickness. We identified the following factors as associated with conversion: male gender, pericholecystic collection seen on ultrasound, gangrenous cholecystitis, and gallbladder wall thickness >1 cm. CONCLUSION: LC is a safe approach in selected patients with acute cholecystitis. Male gender, pericholecystic collection seen on ultrasound, gangrenous cholecystitis, and gallbladder wall thickness >1 cm are associated with a higher risk of conversion to open surgery. PMID- 19130341 TI - [Evaluation of patients with urogenital trauma managed in a urology clinic]. AB - BACKGROUND: Ten percent of all traumas, responsible for 14% of all deaths, involve the urogenital system. We retrospectively evaluated the patients with genitourinary trauma who underwent any kind of management modality in our clinics. METHODS: We retrospectively evaluated 108 patients (92 males, 16 females; mean age 35.8+/-17.5 years; range 6 to 87 years) with urogenital trauma between 2003 and 2007 according to age, gender, type of trauma, affected organ, grade of trauma, accompanying other-organ injuries, radiological imaging techniques, and treatment. RESULTS: Sixty-eight patients (63%) had blunt, 25 (24%) had penetrating, and 12 (11%) had iatrogenic trauma, and 3 patients (3%) had spontaneous organ injury. When we considered the affected organ, renal trauma was determined in 34 patients (32%), ureteral trauma in 11 (10%), bladder injuries in 18 (17%), urethral trauma in 27 (25%), testicular trauma in 6 (6%), and penile trauma in 12 (10%) patients. Seventeen patients (50%) had grade 3, 11 (32%) had grade 4 and 6 (18%) had grade 5 renal injury; 6 patients of these cases underwent surgical treatment. CONCLUSION: Our results were parallel to those available in the literature. Due to the lack of sufficient information about urogenital trauma rates in our country, it would be highly useful for reference centers to evaluate and publish their own data. PMID- 19130342 TI - [Diagnosis that should be remembered during evaluation of trauma patients: diaphragmatic rupture]. AB - BACKGROUND: We aimed to emphasize the importance of delay in diagnosis of traumatic diaphragmatic ruptures and to investigate the results of treatment methods. METHODS: The records of 13 patients (11 males, 2 females; mean age 34.6; range 7 to 52 years) with traumatic diaphragmatic ruptures were evaluated with respect to demographic data, type and time of trauma, symptoms, diagnostic methods, localization, surgical therapy, morbidity and mortality. RESULTS: Blunt trauma was present in 8 cases (61.6%) and penetrating trauma in 5. Migration of abdominal organs into hemithorax was detected in 11 cases radiologically. The duration between diagnosis and the event was 2-20 years in 6 patients (46.1%). Three of them were penetrating and 3 were blunt trauma. In 3 cases, diagnosis had been overlooked despite exploratory thoracotomy and laparotomy in another center. Right hemi-diaphragm was injured in 1 patient and the left in 12 cases. Primary repair through posterolateral thoracotomy was performed in all cases. Morbidity occurred in 1 patient. No mortality was observed. CONCLUSION: Diaphragmatic ruptures should be remembered in trauma cases because of the probability of their being missed, and ruptures should be explored during thoracotomy or laparotomy performed for other organ injuries. Delayed diagnosis will increase morbidity and mortality. PMID- 19130343 TI - [Evaluation of Ohmann and Eskelinen scores, leukocyte count and ultrasonography findings for diagnosis of appendicitis]. AB - BACKGROUND: The purpose of this study was to demonstrate the diagnostic value of Ohmann and Eskelinen scores, leukocyte count and ultrasonographic (USG) evaluations in acute appendicitis. METHODS: This is a retrospective presentation of 186 patients (105 males, 81 females; mean age 29.94+/-12.70; range 6 to 70 years) operated with the diagnosis of acute appendicitis between May 1, 2004 and January 1, 2005. The diagnostic value of Ohmann and Eskelinen scores, leukocyte counts, and USG evaluations and the differences between the sexes were studied. RESULTS: According to the evaluation of Ohmann and Eskelinen scores, there were significant differences in positive and negative appendectomy between the total patient population and female patients (p=0.01), but no significant difference was found for men (p=0.341 and p=0.217, respectively). The leukocyte count increase and USG evaluations also demonstrated significant differences between negative and positive appendectomy in female patients (p=0.001 and p=0.001, respectively). However, no significant differences in the leukocyte count increase and USG evaluations between positive and negative appendectomy were determined in male patients (p=0.480 and p=0.732, respectively). CONCLUSION: Ohmann and Eskelinen scores could be useful in definitive diagnosis of appendicitis especially in primary care units. The leukocyte count increase alone or with the help of USG evaluation could be helpful in the diagnosis of appendicitis especially in females during the reproductive period. PMID- 19130344 TI - [Late-onset traumatic rupture of the diaphragm: two case reports]. AB - Ruptured diaphragm as a result of blunt thorax or abdominal trauma can present acutely or late in the disease. Symptoms may be nonspecific and diagnosis can easily be missed. Patients may present with dyspnea, chest pain or cough. Chest radiograph, CT scan, and MRI are the primary diagnostic tools. Clinicians must have a high index of suspicion for prompt diagnosis, especially in patients with a history of trauma to the abdomen or thorax. The only treatment in diaphragm rupture is surgery. We report two cases of traumatic diaphragm ruptures presenting years after the trauma. PMID- 19130345 TI - [Glomus tumor of the stomach: a rare cause of upper gastrointestinal bleeding]. AB - Glomus tumors are benign soft tissue neoplasms that usually arise in the skin of the extremities; however, those tumors are sometimes located in other sites such as gastrointestinal (GIS), respiratory, and genital tracts. Gastrointestinal glomus tumors are mostly located in the gastric antrum. The most are small (<4 cm), but a few can be enormous. Herein, we report a case of a glomus tumor of the stomach in a 31-year-old female patient who presented with intermittent nausea, vomiting, hematemesis and melena for 2 months. On upper GIS endoscopy, submucosal lesion with ulceration was found in the prepyloric area. Partial gastrectomy including the lesion was performed. Immunohistochemical examination showed the glomus tumor. Since the glomus tumor is essentially benign and does not need a radical surgical procedure, the most important aspect of this tumor is its histologic identification and differentiation from more common gastric lesions, especially from malignant tumors. PMID- 19130346 TI - [Biliary ascaris-induced obstructive jaundice: a case of acute abdomen]. AB - Ascaris lumbricoides is one of the most common helminthic diseases worldwide and it can cause various complications. The presence of this worm in the biliary tree, however, is a rare condition, especially in children. We describe a child with biliary ascariasis who was presenting clinical symptoms mimicking surgical acute abdomen. The diagnosis was made by ultrasound. The child was successfully treated with mebendazole and anti-spasmolytic. Biliary ascariasis should be added to the differential diagnosis of acute abdominal pain in patients, even in children, from endemic areas. PMID- 19130347 TI - Penetrating spinal injury: reports of two cases. AB - Penetrating spinal cord injuries caused by stab wounds are rare. Such injuries may result from a direct lesion of the neural elements of the spinal cord, cord infarcts or, more rarely, intradural or epidural hematoma. In the present study, two cases with spinal cord and L4 root injuries caused by a knife are presented. The first case, a 22-year-old male, referred to our emergency outpatient clinic with a stab wound injury at the mid-section of his back. Neurological examination of this patient showed paraplegia, and spinal MRI displayed a total spinal cord lesion at the T7-T8 level. The wound was primarily sutured; however, during follow-up, CSF (cerebrospinal fluid) leakage continued and the patient was operated. The ruptured dura mater was primarily sutured in a surgical intervention that involved T7-T8 total laminectomy. The second patient referred to our emergency outpatient clinic with a torso injury caused by a sharp knife. The knife was embedded 2-3 cm deep at the wound site at the L3 level and the handle was broken. The patient was immediately operated and the broken and embedded metallic part of the knife was extracted. PMID- 19130348 TI - Spontaneous resolution of a large chronic subdural hematoma: a case report and review of the literature. AB - Spontaneous resolution of a chronic subdural hematoma has been reported rarely in the literature. We present herein the case of a 35-year-old patient with spontaneous resolution of a large chronic subdural hematoma, the volume of which was measured as approximately 76.5 ml on magnetic resonance imaging. No pathology was determined on neurological examination. Neither cerebral angiography nor EEG showed any abnormality. We discuss the possible relation between mechanisms of physio-pathogenesis and spontaneous resolution of chronic subdural hematomas, and also present the patient characteristics together with the other literature data. PMID- 19130349 TI - Detection and treatment of traumatic separation of the distal humeral epiphysis in a neonate: a case report. AB - Traumatic separation of the distal humeral epiphysis during delivery is an uncommon injury in neonates and usually mimics elbow dislocation. Emergency medicine physicians and orthopedic surgeons must have a high index of suspicion for distal humeral epiphysis separation when evaluating elbow trauma in neonates. Magnetic resonance imaging (MRI) scan is an important diagnostic tool for this purpose. We report a case in which fracture-separation of the distal humeral epiphysis in a newborn was diagnosed with the help of ultrasonography and MRI scan, which provided a clear delineation of the injury. PMID- 19130351 TI - Congenital cataract in 1-day-old French Mulard ducklings. AB - Ocular opacity, associated with reluctance to move and inability to feed properly, was observed in approximately 1% of all newly hatched females from several related flocks of Mulard ducks. A 5-week follow-up study of 10 1-day-old affected females was performed, and they were compared with 10 control animals. Clinical, ocular and ultrasonographic examinations, and a complete necropsy of two animals per group with histological examination of the eye, were performed weekly. A bilateral immature cortical anterior cataract was diagnosed at ocular examination and confirmed by ultrasonography in affected ducks. Dyscoria was occasionally observed in affected animals. Severe cataract, with Morgagnian globules, severe anterior fibre liquefaction and disorganization were observed by photonic microscopy. No retinal or choroidal lesions were observed. No progression or repair of ultrasonographic and microscopic lesions could be detected during the 5 weeks of examination. The female predisposition for the ocular lesions suggests a congenital sex-linked recessive cataract. PMID- 19130352 TI - Isolation and identification of highly pathogenic avian influenza H5N1 virus from Houbara bustards (Chlamydotis undulata macqueenii) and contact falcons. AB - Highly pathogenic influenza virus (HPAIV) H5N1 has caused mortality and morbidity in many species of domestic and wild bird. The Houbara bustard (Chlamydotis undulata macqueenii) is a solitary bird that inhabits semi-desert regions. It is known to be susceptible to avianpox, avian paramyxovirus type 1, and low pathogenicity avian influenza H9N2. We report an outbreak of H5N1 HPAIV in Houbara bustards, which were introduced into the Kingdom of Saudi Arabia for falconry purposes. Ninety-three per cent mortality (38 out of 41 birds) in the infected Houbara bustard flock and about 62.5% mortality (10 out of 16 birds) in falcons that came in contact with these birds were observed. Pooled cloacal and tracheal swabs from Houbara bustards as well as visceral organ homogenates collected in Houbara bustards and falcons were tested by real-time reverse transcriptase-polymerase chain reaction, and virus isolation was attempted in specific pathogen free hens' eggs. The viruses isolated were characterized as HPAIV H5N1. Phylogenetic analysis of the haemagglutinating and Neuraminidase (NA) genes revealed that the viruses isolated from Houbara bustards and falcons were closely related to each other and to Kuwaiti H5N1 strains isolated in 2007. Interestingly, they were genetically distinguishable from the co-circulating A/H5N1 viruses in Kingdom of Saudi Arabia causing outbreaks in domestic birds. This case emphasizes the need for surveillance of this endangered species in its natural habitat. PMID- 19130353 TI - Safety-seeking and coping behavior during exposure tasks with anxious youth. AB - This study examined child behavior during exposure tasks and characteristics of the exposure tasks as related to outcomes when treating anxious youth. Participants (aged 7-13) were 87 anxiety-disordered children (37 girls; 50 boys) and their parents (84 mothers; 70 fathers) who completed a 16-session cognitive behavioral therapy. Videotapes of exposure-based sessions were coded for (a) child use of safety-seeking behavior (SSB) and (b) coping behavior (CB) during exposure, (c) characteristics of the exposure tasks, and (d) child anxiety during exposure. Results indicated that child use of SSB, but not CB, during exposure was significantly predictive of poorer outcome. SSB was greater during exposure tasks for treatment nonresponders than for responders, whereas CB was greater for treatment responders. SSB increased over the course of exposure-based sessions, whereas CB remained stable. Child anxiety during exposure tasks and most exposure task characteristics were not significantly predictive of outcome. PMID- 19130354 TI - Contagious depression: negative attachment cognitions as a moderator of the temporal association between parental depression and child depression. AB - This study examined whether negative attachment cognitions moderate the association between the onset of depressive symptoms in children and their parents using a high-risk sample (parents with a history of major depressive episodes and their children) and a multiwave longitudinal design. During the initial assessment, 140 children (ages 6-14) completed a measure assessing parent child attachment cognitions. Parents and children also completed measures assessing current level of depressive symptoms. Following the initial assessment, children and parents were contacted every 6 weeks for the next year to complete measures assessing depressive symptoms. The results of hierarchical linear modeling analyses indicated that children who exhibited high levels of negative attachment cognitions reported greater elevations in depressive symptoms following elevations in their parent's level of depressive symptoms than children who exhibited low levels. PMID- 19130355 TI - Sex differences and response styles: subtypes of rumination and associations with depressive symptoms. AB - In view of recent findings regarding the multifaceted nature of rumination in adults and older adolescents, the purpose of this study was to evaluate the construct of rumination as a 2-factor model (brooding and reflection) in a child and early adolescent sample as well as examine sex differences and associations between depressive symptoms and these ruminative subtypes. Participants were children in 2nd through 7th grades (N = 303). Results of a confirmatory factor analysis supported the hypothesis that subtypes of rumination (2-factor model) are present in children and provided a better fit than a unitary construct of rumination (1-factor model). As predicted, girls endorsed higher levels of brooding than boys, and brooding (not reflection) was uniquely related to depressive symptoms. Results revealed that a distinction between brooding and reflection subtypes is warranted as it can help lead to better assessment and identification of risk factors of depression. PMID- 19130356 TI - Expressed emotion in mothers of currently depressed, remitted, high-risk, and low risk youth: links to child depression status and longitudinal course. AB - This study examined expressed emotion in the families of children and adolescents who were (a) in a current episode of Major Depressive Disorder (MDD), (b) in remission from a past episode of MDD, (c) at high familial risk for developing MDD, and (d) low-risk controls. Participants were 109 mother-child dyads (children ages 8-19). Expressed emotion was assessed using the Five Minute Speech Sample, and psychiatric follow-ups were conducted annually. Mothers of children with a current or remitted episode of MDD and at high risk for MDD were more likely to be rated high on criticism than mothers of controls. There were no differences in critical expressed emotion among mothers of children in the current, remitted, or high-risk for depression groups. Higher initial critical expressed emotion was associated with a greater likelihood of having a future onset of a depressive episode in high-risk and depressed participants. Diagnostic groups did not differ in Emotional Overinvolvement. PMID- 19130358 TI - Adolescent mothers leaving multigenerational households. AB - This study examined how the developmental processes of autonomy and relatedness are related to changes in the residential status of 181 first-time, adolescent, urban, low-income, African American mothers over the first 24 months postpartum. Although adolescent mothers were eager to live independently, few made a clear transition out of the multigenerational household; 56% lived in the household of origin continuously (IN), 21% left and never returned (OUT), and 23% had multiple moves in and out of the household (IN/OUT). Older adolescent maternal age, less supportive adolescent mother-grandmother relations, and high household density were associated with leaving the household of origin. The IN/OUT group had difficulty adopting the roles of adult and parent. Helping adolescent mothers and grandmothers negotiate roles to reduce conflict may promote autonomy and relatedness, allowing mothers to learn parenting skills, qualify for public assistance, and continue their education. PMID- 19130357 TI - Maternal depression, paternal psychopathology, and toddlers' behavior problems. AB - This article examined the effects of maternal depression during the postpartum period (Time 1) on the later behavior problems of toddlers (Time 3) and tested if this relationship was moderated by paternal psychopathology during toddlers' lives and/or mediated by maternal parenting behavior observed during mother-child interaction (Time 2). Of the 101 mothers who participated in this longitudinal study with their toddlers, 51 had never experienced an episode of Major Depressive Disorder (MDD) and 50 had experienced an episode of MDD during the first 18 months of their toddlers' lives. Maternal depression at Time 1 was significantly associated with toddlers' externalizing and internalizing behavior problems only when paternal psychopathology was present. As predicted, maternal negativity at Time 2 was found to mediate the relationship between maternal depression at Time 1 and toddlers' externalizing behavior problems at Time 3. PMID- 19130359 TI - Romantic experience and psychosocial adjustment in middle adolescence. AB - Concurrent and longitudinal relations between the amount of romantic experience and psychosocial adjustment were examined in a 1-year study of a community based sample of 200 tenth graders. Adolescents, parents, and friends completed measures of psychosocial adjustment. The amount of romantic experience was associated with higher reports of social acceptance, friendship competence, and romantic competence; at the same time, romantic experience also was associated with greater substance use, more delinquent behavior, and more frequent genital sexual behavior. The amount of romantic experience predicted increased substance use and genital sexual behavior over a 1-year period, whereas social acceptance predicted increased romantic experience. These relations with romantic experience were significant even when genital sexual behavior and social acceptance were taken into account, underscoring the unique contribution of romantic experiences. PMID- 19130360 TI - Long-term youth criminal outcomes in MST transport: the impact of therapist adherence and organizational climate and structure. AB - This study investigated relations among therapist adherence to an evidence-based treatment for youth with serious antisocial behavior (i.e., Multisystemic Therapy), organizational climate and structure, and youth criminal charges on average 4 years posttreatment. Participants were 1,979 youth and families treated by 429 therapists across 45 provider organizations. Results showed therapist adherence predicted significantly lower rates of youth criminal charges independently and in the presence of organizational variables. Therapist perceptions of job satisfaction and opportunities for growth and advancement relative to the organizational average predicted youth criminal charges, as did organizational average levels of participation in decision making. These associations washed out in the presence of adherence, despite the fact that job satisfaction and growth and advancement were associated with adherence. PMID- 19130361 TI - Adolescents' physical aggression toward parents in a clinic-referred sample. AB - Physical aggression directed toward parents by their adolescents is a serious issue both practically and scientifically. In contrast to the extensive literature on other forms of aggression within families (e.g., marital violence, child physical abuse) as well as youth aggression construed broadly, a major gap exists in our knowledge of youth-to-parent physical aggression (YPA). In this study, we analyzed data on three forms of physical aggression (YPA, interparental, and parent-to-youth) from 232 mother-adolescent dyads drawn from a database of families referred for the clinical treatment of emotional and behavioral problems in their adolescent children. Analyses indicated that YPA is prevalent (57% by sons and 49% by daughters in 1 year) and significantly likely to co-occur with interparental and parent-to-youth aggression in the family. Follow-up analyses suggested important sex differences in these relations. PMID- 19130362 TI - Relations between parental psychological control and childhood relational aggression: reciprocal in nature? AB - Using a cross-lagged panel design, this study examined the directionality of relations between parental psychological control and child relational aggression. Data were collected from a proportionally stratified sample of 600 Flemish 8- to 10-year-old children at 3 measurement points with 1-year intervals. Reciprocal effects were evident in mother-child dyads. Maternal psychological control was positively related to child relational aggression 1 year later, and child relational aggression was positively related to maternal psychological control 1 year later. The father-child dyads were best represented by unidirectional parent effects. Paternal psychological control was positively related to child relational aggression 1 year later. Surprisingly, these effects emerged only for relational aggression as indexed by mother and father reports and not for relational aggression as indexed by teacher and peer measures. PMID- 19130364 TI - Applications of generalizability theory to clinical child and adolescent psychology research. AB - Using generalizability theory to evaluate the reliability of child and adolescent measures enables researchers to enhance precision of measurement and consequently increase confidence in research findings. With an observer-rated measure of child self-regulation, we illustrate how multiple sources of error variance (e.g., raters, items) affect the dependability (replicability) of scores and demonstrate methods for enhancing dependability of observer ratings. Using ratings of 181 children, we illustrate the use of two-facet (i.e., raters and items as sources of error) and three-facet (i.e., raters, items and occasions) analyses to optimize design features of future studies using this measure. In addition, we show how generalizability theory provides a useful conceptual framework for thinking about determinants of scores on acquaintance (e.g., teacher or parent) ratings, as well as observer ratings, and sheds light on the strengths and limitations of both types of data for child and adolescent clinical research. PMID- 19130363 TI - Few girls with childhood attention-deficit/hyperactivity disorder show positive adjustment during adolescence. AB - Employing data from 140 prospectively followed girls with attention deficit/hyperactivity disorder (ADHD) and 88 age- and ethnicity-matched comparison girls, we adopted a person-centered analytic approach to assess rates of adolescent positive adjustment (PA) across six domains: ADHD symptoms, externalizing symptoms, internalizing symptoms, social skills, peer acceptance, and school achievement. During adolescence, between 19.8% and 61.1% of the girls with childhood ADHD met criteria for PA when the six domains were considered independently. A total of 16.4% of the ADHD sample showed PA in at least five of six domains, versus 86.4% of the comparison girls. Results were similar when PA was examined excluding the ADHD symptom domain. Most girls did not "grow out of" the symptoms and impairments related to their ADHD. PMID- 19130365 TI - Validation of the preschool and primary school form of a questionnaire assessing parents' childrearing behavior. AB - This study presents a validation of a scale that assesses parents' childrearing behavior toward young children. The scale was validated on 565 parents of 2- to 7 year-old children. The current results replicated the factor solution of the original scale designed for parents of school-aged children. The scale demonstrated good psychometric properties: moderate to high internal consistency, the expected relations with criterion variables (parental self-efficacy beliefs, child's behavior and personality), and discriminative properties according to the parents' gender and educational level, the child's age and gender, and the difference between referred and nonreferred children. PMID- 19130367 TI - Toxicity of lindane (gamma-hexachloroxiclohexane) in Sparus aurata, Crassostrea angulata and Scrobicularia plana. AB - The purpose of the present study was to research the sublethal and/or lethal effects produced by the exposure of fish and shellfish to the gamma isomer of lindane, gamma-hexachlorocyclohexane (gamma-HCH). The teleostean fish Sparus aurata and the shellfish Crassostrea angulata and Scrobicularia plana, were exposed to 16 micro g/L of lindane for 15 days. Samples of different fish (liver, kidney and gills) and shellfish (gills, gut, digestive gland and mantle) tissues were extracted and processed for histopathological observations. Although mortality was not detected during the bioassay, sublethal effects (histopathological alterations) were observed. Vacuolization in the liver cells and lamellar fusion in gills from exposed fish were observed. Disorganization of normal gill structure, epithelial desquamation with the disappearance of apical ciliature in intestine, and inflammatory response in mantle from exposed shellfish were also observed. Thus, it can be concluded that the lindane concentration employed in the present research did not produce lethal effects in the exposed organisms but it caused sublethal effects. Lindane has time-dependent multiple toxic effects in S. aurata, C. angulata and S. plana, which were more severe at the end of the experimental time. The toxicological implications arising from these results are subjects for further multiconcentration tests dealing with lethal responses (mortality) or with sublethal responses (cellular/molecular biomarkers) of the aforementioned species. PMID- 19130366 TI - Family-based treatment of a 17-year-old twin presenting with emerging anorexia nervosa: a case study using the "Maudsley method". AB - This article describes the successful application of family-based treatment (FBT) for a 17-year-old identical twin presenting with a 4-month history of clinically significant symptoms of anorexia nervosa (AN). FBT is a manualized treatment that has been studied in randomized controlled trials for adolescents with AN. This case study illustrates the administration of this evidence-based intervention in a clinical setting, highlighting how the best available research was used to make clinical decisions at each stage of treatment delivery. PMID- 19130368 TI - Influence of avian reproduction ecotoxicological endpoints in the assessment of plant protection products. AB - The aim of this paper is to examine the statistical relevance of bird species on the endpoints of avian long-term toxicity studies (eggs laid, eggs set, eggs hatching, embryo survivor, 14-day old survivors and eggshell thickness). Data from 561 animals of three different species (Colinus virginianus, Anas platyrynchos and Coturnix coturnix japonica) tested with five different pesticides were analyzed in this study. The substances considered were: Thiamethoxam (EZ-3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4 ylidene(nitro)amine), Thiacloprid ((Z)-3-(6-chloro-3-pyridylmethyl)-1,3 thiazolidin-2-ylidenecyanamide), Acetamiprid ((E)-N(1)-[(6-chloro-3 pyridyl)methyl]-N(2)-cyano-N(1)-methylacetamidine), Phosmet (O,O-dimethyl S phthalimidomethyl phosphorodithioate) and Dicofol (2,2,2-trichloro-1,1-bis(4 chlorophenyl)ethanol). Several general lineal mixed models were conducted to evaluate the factors affecting variables used in long-term reproductive toxicity tests. Test significance was p < 0.01 in all models tested. Model R(2) value was high (0.80) for all variables except for eggs laid (R(2) = 0.42) for the three species studied. Tukey studentized range test showed significant differences among species and pesticides. For pre-hatching period the differences were significant for eggs laid and eggs set among species. C. japonica showed statistical differences for egg hatching. With respect to embryo survivor and 14 days old survival, significant different were found for C.virginianus and A. platyrynchos, respectively. These results indicate that the selected species have an influence in the endpoints to be used for risk assessment. PMID- 19130369 TI - The controlled and sustained release of a fungicide from starch and alginate beads. AB - This paper describes the synthesis of starch and alginate beads as controlled release formulations. Different formulations of the fungicide, thiram, were prepared by using BaCl(2) and AlCl(3) as crosslinkers. The beads thus formed were characterized by FTIR and swelling studies. Formulation characteristics like entrapment efficiency, bead size, percentage equilibrium swelling have been evaluated. The release studies of the fungicide from the beads were carried out in vitro and in soil columns. The beads prepared with AlCl(3) have shown more swelling as well as the fungicide release in comparison to beads prepared with BaCl(2). From the soil column studies it was observed that the release of pesticide from the beads occurred in very controlled and sustained manner. PMID- 19130370 TI - Persistence of the herbicide butachlor in soil after repeated applications and its effects on soil microbial functional diversity. AB - Effects of repeated applications of the herbicide butachlor (N-(butoxymethyl)-2 chloro -N-2',6'-dimethyl acetanilide) in soil on its persistence and soil microbial functional diversity were investigated under laboratory conditions. The degradation half-lives of butachlor at the recommended dosage in soil were calculated to be 12.5, 4.5, and 3.2 days for the first, second, and third applications, respectively. Throughout this study, no significant inhibition of the Shannon-Wiener index H' was observed. However, the Simpson index 1/D and McIntosh index U were significantly reduced (P < or = 0.05) during the initial 3 days after the first application of butachlor, and thereafter gradually recovered to a similar level to that of the control soil. A similar variation but faster recovery in 1/D and U was observed after the second and third Butachlor applications. Therefore, repeated applications of butachlor led to more rapid degradation of the herbicide, and more rapid recovery of soil microorganisms. It is concluded that repeated butachlor applications in soil had a temporary or short-term inhibitory effect on soil microbial communities. PMID- 19130371 TI - Persistence and distribution of pesticide residues in fresh agricultural food consumed in the province of Bologna. AB - The presence of pesticide residues in fresh vegetables and fruit have been qualitatively and quantitatively determined at the laboratories of the Regional Agency for Environmental Protection (ARPA), Division of the Province of Bologna. More than 1,700 samples have been tested by routine analyses. The possible risks for consumers have been evaluated by various parameters. The most important ones were: the amount of each residue; the respective ADI (Acceptable Daily Intake) limit; the contemporary presence of different residues; an estimation of the daily intake, based on the amount of fruit and vegetables consumed per person. It has been possible to evaluate that the daily intake of pesticide residues in the province of Bologna during the period 2003-06 resulted lower than the ADI limits concerning the vegetables. According to the information on fruit consumption the daily intake of omethoate (O,O-dimethyl S-methylcarbamoylmethyl phosphorothioate) resulted higher than its ADI limit, of dicofol (2,2,2-trichloro-1,1 bis(trichloromethyl)benzenemethanol) very close to the admitted limit, under the respective limits for all the other residues. PMID- 19130372 TI - Evaluation of the field dissipation of fungicides and insecticides used on fruit bearing trees in northern Italy. AB - The dissipation of the fungicides captan, cyprodinil, fludioxonil, dithianon, and tebuconazole and of the insecticides chlorpyrifos, fenitrothion, and malathion was studied, following a single treatment of different cultivars of pears, apples, and peaches. The study was conducted in northern Italy, over two successive growing seasons (2004 and 2005). The treatments were performed by the farmers involved, in line with their usual practice. At various time intervals from treatment to harvest, representative samples of fruit were collected and analyzed for pesticide residues. In some cases, concentrations lower than the maximum residue levels (MRLs) were found immediately after treatment. In all trials a rapid decline in pesticide concentrations was observed leading to residues at harvest greatly below the MRLs. PMID- 19130373 TI - Effect of a subchronic exposure to simazine on energetic metabolism of common carp (Cyprinus carpio). AB - The energetic parameters, such as glycogen, glucose, proteins, lactate and adenosine triphosphate (ATP) content and lactate deshydrogenase (LDH) activity in tissues and blood of carps from simazine (2-chloro-4,6-bis(ethylamino)-s triazine) contaminated waters were investigated. In order to confirm the field results, a laboratory experiment was designed in which carps were exposed to simazine at the concentration level of 45 micro g. L(- 1) (10-fold of the amount found in natural waters) for 90 days. Fish from a contaminated reservoir showed low glycogen concentrations in hepatopancreas and muscle, while fish in another contaminated reservoir showed high LDH activity together with an increase in lactate content in muscle. Laboratory findings did not confirm field results, and fish exposed to simazine did not show alterations in the parameters studied. The results suggest that carps were not stressed by the presence of the simazine at the concentration levels found in both studies and the mechanisms of defense covered the energetic demand. PMID- 19130374 TI - Acute toxic effects of three pesticides on Pseudomonas putida monitored by microcalorimeter. AB - A series of calorimetric experiments were performed to investigate the toxic effects of beta-cypermethrin (BCP), bensulfuron-methyl (BSM) and prometryne (PM) on Pseudomonas putida (P. putida). The metabolic action of P. putida on the three pesticides was studied by obtaining power-time curves. The growth of P. putida was inhibited completely in each case when the concentrations of pesticides were up to 80 micro g mL(- 1). The relationships between the inhibitory ratio (k) and doses of contaminants were approximately linear for the three pesticides. The total heat dissipated per milliliter (Q(total)) for the pesticides decreased during the course of the experiment. The OD(600) of P. putida growth in the absence and presence of pesticides was also obtained. The power-time curves of P. putida growth coincided with its turbidity curves. This elucidates that microcalorimetric method agrees well with the routine microbiological method. Among these three pesticides, BSM was found to be the most toxic with an IC(50) of 19.24 micro g mL(- 1) against P. putida. PM exhibited moderate virulence with an IC(50) of 27.86 micro g mL(- 1) and BCP had the lowest toxicity with an IC(50) of 39.64 micro g mL(- 1). PMID- 19130375 TI - Mercaptobenzothiazole-on-gold organic phase biosensor systems: 1. Enhanced organosphosphate pesticide determination. AB - This paper reports the construction of the gold/mercaptobenzothiazole/polyaniline/acetylcholinesterase/polyvinylacetate (Au/ MBT/PANI/AChE/PVAc) thick-film biosensor for the determination of certain organophosphate pesticide solutions in selected aqueous organic solvent solutions. The Au/MBT/PANI/AChE/PVAc electrocatalytic biosensor device was constructed by encapsulating acetylcholinesterase (AChE) enzyme in the PANI polymer composite, followed by the coating of poly(vinyl acetate) (PVAc) on top to secure the biosensor film from disintegration in the organic solvents evaluated. The electroactive substrate called acetylthiocholine (ATCh) was employed to provide the movement of electrons in the amperometric biosensor. The voltammetric results have shown that the current shifts more anodically as the Au/MBT/PANI/AChE/PVAc biosensor responded to successive acetylthiocholine (ATCh) substrate addition under anaerobic conditions in 0.1 M phosphate buffer, KCl (pH 7.2) solution and aqueous organic solvent solutions. For the Au/MBT/PANI/AChE/PVAc biosensor, various performance and stability parameters were evaluated. These factors include the optimal enzyme loading, effect of pH, long-term stability of the biosensor, temperature stability of the biosensor, the effect of polar organic solvents, and the effect of non-polar organic solvents on the amperometric behavior of the biosensor. The biosensor was then applied to detect a series of 5 organophosphorous pesticides in aqueous organic solvents and the pesticides studied were parathion-methyl, malathion and chlorpyrifos. The results obtained have shown that the detection limit values for the individual pesticides were 1.332 nM (parathion-methyl), 0.189 nM (malathion), 0.018 nM (chlorpyrifos). PMID- 19130376 TI - Pungency in Capsicum chinense: variation among countries of origin. AB - Fruits of 63 accessions of Capsicum chinense Jacq. from the USDA/ARS Capsicum germplasm collection were analyzed for two major capsaicinoids, capsaicin and dihydrocapsaicin, using gas chromatography with nitrogen phosphorus detection (GC/NPD). The objectives of the present investigation were: (i) to quantify the major capsaicinoids (capsaicin and dihydrocapsaicin) in fruits of Capsicum chinense accessions and (ii) to identify accessions containing great concentrations of capsaicinoids among countries of hot pepper origin. Seeds of C. chinense accessions received from Belize, Brazil, Colombia, Ecuador, Mexico, Peru, Puerto Rico, and United States were field grown in a silty-loam soil. Mature fruits were analyzed for major capsaicinoids content. Capsaicin concentrations were generally greater than dihydrocapsaicin. Fruits of C. chinense accession PI640900 (USA) contained the greatest concentration of capsaicin (1.52 mg g(- 1) fruit) and dihydrocapsaicin (1.16 mg g(- 1) fruit), while total major capsaicinoids in the fruits of PI438648 (Mexico) averaged 2 mg g(- 1) fruit. These two accessions were identified as potential candidates for mass production of major capsaicinoids that have health-promoting properties and for use as a source of pest control agents in agricultural fields. PMID- 19130377 TI - Nutrient release from fish silage using microwave-enhanced advanced oxidation process. AB - The microwave-enhanced advanced oxidation process was used to treat fish silage for nutrient release and solids reduction prior to its use as a fertilizer for greenhouse operations. Fifteen sets of experiments with varying hydrogen peroxide dosages and treatment temperatures were conducted to evaluate the effectiveness of the process on the solubilization of fertilizer constituents. It was found that up to 26% of total Kjeldahl nitrogen could be released as ammonia with 6% hydrogen peroxide dosage at 170 degrees C. An increase of nitrate/nitrite concentration was observed with higher hydrogen peroxide dosage and higher microwave temperature; the highest concentration of 10.2 mg L(- 1) nitrates/nitrites was achieved at at 170 degrees C and 6% H(2)O(2) dosage. Up to 20 +/- 9.5% of total chemical oxygen demand was reduced at temperatures between 120 and 170 degrees C. Large quantities of volatile fatty acids were generated at lower temperatures, corresponding to an increase in soluble chemical oxygen demand, but not at higher temperatures. The treatment of fish silage using the microwave-enhanced advanced oxidation process appears to be promising. PMID- 19130378 TI - Insecticide residues on hands: assessment and modeling with video observations of determinants of exposure--a study among subsistence farmers in nicaragua. AB - This investigation quantitatively assessed hand residues of chlorpyrifos and methamidophos in a field setting and sought to explain the residues through application volume and determinants of exposure using application data for 28 subsistence farmers in the Pacific Region of Nicaragua. Hand residues were estimated by recovery of the pesticides by standardized wipe sampling for both hands, analyzed with solvent extraction and gas chromatography with electron capture detector. Application volumes were based on data on individual spraying rates and mixing volumes. Eleven determinants of exposure, related to work practices during mixing and spraying of the pesticides, were assessed for each subject from videotapes. Correlation and regression analyses estimated the associations between hand residues, application volume, pesticide type, and determinants of exposure. Correlations between residues for different hand parts were high (r 0.75-0.98). Total hand residue (sum of residues of parts of both hands) correlated with application volume (r 0.43, p 0.02), not washing hands (r 0.41, p 0.04), spraying nozzle forward (r 0.26, p 0.17), manipulation of hose (r 0.32, p .09), and insecticide type (chlorpyrifos vs. methamidophos; r 0.31, p 0.10). A model that explained total hand residue with these five variables yielded a multiple correlation coefficient of 0.67 (p 0.01). Unmeasured determinants and/or narrow range of the exposure situation probably account for the unexplained variance of the residues. PMID- 19130379 TI - Who gets "kicked out" of the exam room? Factors associated with patients declining medical student participation. AB - BACKGROUND: Research on patients' decision to have medical students participate in clinical care has been examined predominantly from the patients' perspective rather than the students' perspective. PURPOSE: This study investigated 3rd-year medical students' experiences with and perceptions of patients declining medical student participation in clinical care and identified factors associated with this decision. METHODS: Third-year medical students (n = 103) completed a survey assessing demographic background, experiences with patients declining medical student participation in clinical care, preceptors' approach to obtaining patient consent for student participation, and ratings of preceptors' clinical education skills. RESULTS: Fifty-nine percent, 22%, and 25% of students encountered patients declining student involvement in care (3 times or more) based on student's training status, student's gender, and in the context of a gynecological/urological exam, respectively. Patient consent obtained by clinic staff (r =.661, p < .001) was a significant predictor for patients declining based on training status. Gender (d = 40, p < .003) and patient consent obtained by clinic staff (r = .349, p < .001) were predictors of patients declining in the context of a gynecological/urological exam. CONCLUSIONS: A majority of medical students have encountered multiple experiences with patients declining student participation in outpatient clinical care. The approach preceptors use to obtain patient consent may be important in increasing training opportunities for students. PMID- 19130380 TI - The patient safety OSCE for PGY-1 residents: a centralized response to the challenge of culture change. AB - BACKGROUND: Accreditation and Institute of Medicine mandates require retooling of graduate medical education curriculum and assessment processes. This Objective Structured Clinical Exam (OSCE) focused on patient safety-specific skills important to stakeholders from multiple institutions. PURPOSES: A 10-station OSCE was designed to assess patient safety-related competencies in new Postgraduate Year 1 (PGY-1) residents. The OSCE emphasized performance of essential skills and teamwork, and it provided early formative feedback to trainees and leadership. METHODS: Group nominal process selected 10 final OSCE stations. Two stations were designed to assess team competencies and response to feedback. Two hundred thirty five trainees enrolled in 64 programs participated during summer 2006. Skill-set aggregation was employed to improve the validity of individual feedback. RESULTS: Significant performance deficits were noted. Trainee and administrator evaluation of the experience was positive. CONCLUSIONS: Multi-institutional test development and centralized testing was well received and produced worrisome results. Early assessment can guide the development of task-specific personalized learning plans and systemwide curricular improvement. Further research is needed to determine whether such an effort directed at PGY-1 trainees can improve trainee performance and patient safety. PMID- 19130381 TI - The effect of monthly resident reflection on achieving rotation goals. AB - BACKGROUND: Reflection is a learning tool increasingly being used in medical education. It has not been well studied as a method of communicating resident rotation goals and objectives. PURPOSE: The purpose was to study the effect of monthly resident reflection on achieving curriculum goals. METHODS: During the first half of the 2005-06 academic year, obstetrics/gynecology residents at a community hospital received curriculum goals in a traditional written manner. During the second 6 months, the same trainees underwent 1-hr monthly structured reflection sessions on the rotation goals and objectives. RESULTS: Sixteen residents were studied. Compared to control, residents reported the rotation goals as better defined during reflection months. More trainees rated reflection as more valuable than the didactic time it replaced. All 16 residents documented more cumulative procedures in the Accreditation Council for Graduate Medical Education operative log Web site during reflection months for each of the three major categories; there was a mean increased number of procedures logged in Obstetrics (23.1 +/- 14.0, p < .001), Gynecology (18.3 +/- 10.9, p < .001), and Primary Care (21.1 +/- 23.5, p = .003). Overall, there was a mean of 62.6 +/- 32.1 (p < .001) more procedures entered during the reflection period, which translated to an 18% increase. CONCLUSIONS: Structured monthly reflection may lead to a higher success of achieving rotation goals. PMID- 19130382 TI - Performance outcomes in anesthesiology residents completing categorical (anesthesia) or advanced (nonspecific) internship training. AB - BACKGROUND: The internship or first year (PGY 1) of anesthesiology training may be categorical (within anesthesiology), or obtained in more diverse settings. Revisions recently proposed in the training requirements incorporated the PGY 1 into the existing curriculum. PURPOSES: We studied whether this change improved measurable outcomes. METHODS: There were 518 residents studied retrospectively from four institutions that offered entry following both "Categorical" and "Other" internships. Thus the training in clinical anesthesia was identical. RESULTS: No differences were observed in percentile scores on the Anesthesiology In-Service Training Examination during clinical anesthesia training, the receipt of awards, board certification or time to certification, or in reports of unsatisfactory performance to the American Board of Anesthesiology. "Categorical" residents were more frequently appointed chief resident. CONCLUSIONS: Easily accessible performance measures may function as valuable aids in decision making, particularly when significant changes in curricula are contemplated. Data do not support the proposed changes in anesthesiology. PMID- 19130383 TI - The impact of leadership and team behavior on standard of care delivered during human patient simulation: a pilot study for undergraduate medical students. AB - BACKGROUND: Methodology to train team behavior during simulation has received increased attention, but standard performance measures are lacking, especially at the undergraduate level. PURPOSES: Our purposes were to develop a reliable team behavior measurement tool and explore the relationship between team behavior and the delivery of an appropriate standard of care specific to the simulated case. METHODS: Authors developed a unique team measurement tool based on previous work. Trainees participated in a simulated event involving the presentation of acute dyspnea. Performance was rated by separate raters using the team behavior measurement tool. Interrater reliability was assessed. The relationship between team behavior and the standard of care delivered was explored. RESULTS: The instrument proved to be reliable for this case and group of raters. Team behaviors had a positive relationship with the standard of medical care delivered specific to the simulated case. CONCLUSIONS: The methods used provide a possible method for training and assessing team performance during simulation. PMID- 19130384 TI - Implementing smokeless tobacco instruction into medical student education: addressing the gap. AB - BACKGROUND: Despite the unique health and epidemiological aspects of smokeless tobacco use, medical education regarding this topic is virtually lacking. DESCRIPTION: The purpose of this study is to develop a comprehensive tobacco curriculum that includes smokeless tobacco education. A thorough review of the literature was carried out to develop includes 8 modules in basic and clinical sciences that are evaluated by pretest/posttest increases in knowledge as well as standardized patient encounters and process evaluation. EVALUATION: Pretest/posttest data indicate that students increased knowledge on specific smokeless tobacco questions. Students also scored well on interactions with standardized patients using the Tobacco Intervention Risk Factor Interview Scale, a validated instrument to assess medical students' tobacco counseling skills. Process evaluation data indicate that modules were generally well received. CONCLUSIONS: This Web-based, comprehensive curriculum-the only curriculum we are aware of treating the topic of smokeless tobacco use-appears to be effective and well received. Smokeless tobacco should be included in medical education. PMID- 19130385 TI - A personal computer-based undergraduate medical school curriculum using SOLE. AB - BACKGROUND: We describe our experience with a personal computer and Web-based undergraduate curriculum for preclinical medical students using the Secure Online Environment (SOLE) education and information system. DESCRIPTION: To test the potential effectiveness of SOLE as a learning tool, we analyzed the patterns of SOLE usage, usage intensity, and consistency among medical students in two preclinical courses (4th-year Human Function and 5th-year Pathology) and attempted to link these indicators to academic performance. Categories of SOLE usage included number of website log-ins and number of pages viewed per course. EVALUATION: We found that A- and B-grade 4th-year students accessed course materials more frequently than did C- to failing-grade students, and both median and mean number of SOLE log-ins declined as student performance decreased. Higher graded students were also more consistent in their usage of SOLE than were lower graded students. The range of log-in numbers (variability in frequency of usage) was greater for C- to failing-grade students than for A- and B-grade students. Compared to their 4th-year counterparts, 5th-year students increased their intensity of SOLE usage (indicated by the number of pages viewed) dramatically and numbers were comparable for A- and B-grade and for C- to failing-grade students. Consistency of usage, however, still remained higher for better performing students. Furthermore, students preferred SOLE to a traditional paper based curriculum and felt it improved teaching effectiveness. Based on usage data and student preferences and perceptions we found Web-based SOLE to be an effective and well-accepted educational tool for preclinical medical students. CONCLUSIONS: This integrative, online educational and information system offers numerous opportunities and advantages for self-assisted instruction that can serve as a foundation for clinical training and professional lifelong learning. PMID- 19130386 TI - Using a novel small-group approach to enhance feedback skills for community-based teachers. AB - BACKGROUND: As medical education expands into distant settings, challenges in providing faculty development to busy clinical teachers increase-especially for those who have difficulty accessing sessions offered at academic centers. DESCRIPTION: Sixty-five clinical teachers participated in six small-group workshops, using a printed module on the topic of delivering feedback. The modules included teaching-learning "cases," tools, and a summary of medical literature. The group facilitator did not require expertise in delivering feedback. Surveys inquired about impact immediately after the session and at 3 months. EVALUATION: Analysis confirmed that participants found the workshop format valuable, and the majority committed to making changes in their approaches to providing feedback. At follow-up, most participants reported that planned changes had been implemented. CONCLUSIONS: A low-tech approach to faculty development, using facilitated small-group discussion of a specially prepared educational module, is feasible for any site and can enhance teaching approaches in both urban and rural practice settings. PMID- 19130388 TI - An Internet guide to key variables for a coherent educational system based on principles of Problem-Based Learning. AB - PURPOSE: It was decided to explore the variables that would need to be considered when planning, revising, or evaluating the outcomes of a whole curriculum, based on the principles of Problem-Based Learning. SUMMARY: A set of major variables was identified at a workshop discussion with academics from medical, dental, and veterinary science faculties. These variables were subsequently reviewed and described by a small group of colleagues in Great Britain, with additional assistance from colleagues in different parts of the world. CONCLUSIONS: The descriptions of 13 variables has been assembled, primarily with a focus on medical education. This guide has been published on the Internet, in order to solicit comments and suggestions from teachers in medicine, dentistry, and veterinary science across the world with the intention of producing a more universally applicable second edition of the guide. PMID- 19130387 TI - Designing a patient safety undergraduate medical curriculum: the Telluride Interdisciplinary Roundtable experience. AB - PURPOSE: Patient safety has emerged as a global concern in the provision of quality health care, and yet, to date, few medical schools have created and/or implemented patient safety curricula. The purpose of this article is to introduce readers to one model of a patient safety undergraduate medical curriculum, as designed by a group of experts attending an annual interdisciplinary roundtable assembled for this purpose. SUMMARY: The Annual Telluride Interdisciplinary Roundtable met in 2005 and 2006 to design what it considered to be a comprehensive patient safety curriculum for medical students. Invited members included stakeholders from a variety of fields, including health care providers, senior health care administration, students, residents, patient advocacy leaders, and curriculum development/assessment experts. The group developed a list of general curricular principles, followed by 11 specific elements felt to be essential to an effective patient safety curriculum for undergraduate medical education students. It also identified a number of challenges to implementing such a curriculum. CONCLUSIONS: A patient safety curriculum, developed by a group of experts for an undergraduate medical education population, was successfully developed over a two-year period of time. Future meetings of the Telluride Roundtable group have centered on evaluation and refinement of these curricular elements as pilots occur in a number of medical schools, and new curricular ideas continue to be developed. Continued interprofessional dialogue and collaborative research will enable the development and implementation of a standardized longitudinal patient safety student curriculum. PMID- 19130389 TI - Teaching histology and histopathology as a surgical registrar in Bulawayo, Zimbabwe. AB - BACKGROUND: Medical schools in the developing world have huge shortages in teaching personnel. The medical school in Bulawayo, Zimbabwe, is educating medical students to become doctors in a country facing huge economic hardships and with a health care system in crisis. SUMMARY: I traveled to Bulawayo to work as a surgical registrar and taught histology and histopathology in the local medical school. Using the local hospital libraries and Internet resources and recruiting busy clinicians in Bulawayo to teach, we were able to set up the first Histopathology course taught in the medical school. The curriculum was decided by local clinicians and emphasized diseases that are prevalent in Zimbabwe. CONCLUSIONS: In an environment of a lack of university lecturers, local clinicians may be an excellent teaching resource. Teaching sessions and locations may be slightly unorthodox but basic science teaching can be delivered in clinical contexts that highlight the clinical relevance of factual information. PMID- 19130391 TI - Journal Watch From ACE (Alliance For Clinical Education): annual review of medical education articles in internal medicine journals 2006-2007. PMID- 19130393 TI - On the determination of ventilatory threshold and respiratory compensation point via respiratory frequency. AB - This study examined the validity of a quantitative respiratory frequency (f(R)) analysis to detect the ventilatory threshold (theta(Vent)) and respiratory compensation point (RCP). Thirty-six amateur competitive cyclists completed a maximal graded exercise test on an electromagnetically-braked cycle ergometer. theta(Vent) and RCP were determined using multiple gas exchange criteria and by f(R) analysis (theta(Vent)f(R) and RCP(f(R))), employing an iterative least squares linear regression technique. Fifteen subjects were excluded from the analyses due to a low signal-to-noise ratio and/or high risk for pseudo-threshold resulting from hyperventilation early in the exercise protocol. A Bland-Altman procedure for inter-analysis comparison completed on the remaining participants' data (n=21; age=29+/-7 years; height=177+/-9 cm; weight=76.0+/-15.8 kg; VO(2max)=4.415+/-0.971 lmin(-1); 58.7+/-10.7 ml kg(-1) min(-1)) revealed mean bias+/-95% Limits of Agreement (LOA) of 1.53+/-50.2 W for theta(Vent) and theta(Vent)f(R). The same inter-anlysis comparison (n=21) for RCP and RCP(f(R)) resulted in a mean bias+/-LOA of 12.6+/-26.9 W. The analysis techniques in the present investigation revealed substantial limits of agreement and/or bias for all estimations, and these data indicated f(R) analyses were unsatisfactory to determine theta(Vent) and RCP in trained cyclists. PMID- 19130394 TI - Aerobic and anaerobic power in relation to age and physical activity in 354 men aged 20-88 years. AB - We examined simultaneously the influence of age as well as both current and historical habitual physical activity (PA) on aerobic (physical working capacity PWC) and anaerobic (Pmax) power in 354 men aged 20.2-87.6 yrs. All the participants performed the graded aerobic exercise test and 8-second explosive power tests on a bicycle ergometer. Current leisure time physical activity (LTPA) and lifetime historical LTPA were assessed. The decline in Pmax/kg (10.3% per decade) was higher than in PWC/kg (7.5% per decade). Correlation coefficients for current LTPA and aerobic indices were higher than for current LTPA and anaerobic indices. In a multiple stepwise regression, current LTPA had the main effect on PWC/kg, whereas age had the main effect on Pmax/kg. Aerobic power was related to all measures of historical PA questionnaire, while anaerobic power - only to those historical measures which pertained to adolescence and younger years of adulthood. We conclude that the age-associated decline in anaerobic power is steeper than that of aerobic power. Habitual endurance-type LTPA increases aerobic but not anaerobic fitness throughout the adult lifespan. Anaerobic fitness indices are higher in young active vs sedentary men but this difference is not discernible as early as in the middle age. PMID- 19130395 TI - [Shwachman-diamond syndrome as cause of infantile eczema associated with failure to thrive]. AB - BACKGROUND: Shwachman-Diamond syndrome is an autosomal recessive multisystem disorder involving an insufficiency of the exocrine pancreas and haematological problems as main symptoms. Frequently, ichthyosiform skin lesions are described but are usually not the leading symptom of the disease. CASE REPORT: We report on a 6-months-old boy suffering from ichthyosiform and eczematous skin eruptions beginning at the age of 3 months which were accompanied by failure to thrive. Because of an atopic predisposition and a sensitisation to hen's egg the diagnosis atopic dermatitis with food allergy was established. We describe the steps leading to the diagnosis Shwachman-Diamond syndrome. CONCLUSION: Shwachman Diamond syndrome may present with skin eruptions as main symptom. A mixed clinical picture with an atopic dermatitis may occur and can aggravate skin symptoms. Additional medical problems like failure to thrive or neutropenia should lead to further diagnostic procedures to exclude Shwachman-Diamond syndrome. PMID- 19130396 TI - Expression of enzymes involved in synthesis and metabolism of estradiol in human breast as studied by immunocytochemistry and in situ hybridization. AB - It is well documented that human breast is actively involved in the local formation of estrogens. To determine the site(s) of action of enzymes involved in synthesis and metabolism of the most potent estrogen estradiol (E2), we have studied the expression of the following enzymes: 3beta-hydroxysteroid dehydrogenase (3-HSD), 17beta-HSD types 1, 2, 5, 7 and 12, aromatase, steroid sulfatase (STS) and estrogen sulfotransferase (EST) 1E1 at the cellular level in breast. Both in situ hybridization and immunocytochemistry were used for enzyme localization in normal breast tissues. For immunocytochemistry, we used rabbit antibodies, while in situ hybridization studies were performed using (35S) labeled cRNA probes. Similar results were obtained with both approaches. All the enzymes (3beta-HSD; 17beta-HSD types 1, 5, 7 and 12; aromatase) involved in the conversion of circulating dehydroepiandrosterone (DHEA) to E2 as well as STS which converts estradiol sulfate (E2-S) to E2 have been found to be expressed in epithelial cells of acini and/or ducts as well as the stromal cells. Moreover, 17beta-HSD type 2 and EST1E1, two enzymes which inactivate E2, have been also localized in the same cell types. The present results indicate the enzymes which play a role in the synthesis and metabolism of E2 are expressed in both epithelial and stromal cells in human breast. PMID- 19130397 TI - Intestinal uptake of amyloid beta protein through columnar epithelial cells in suckling mice. AB - The mechanism of transmission of amyloid protein, especially the dynamics in the intestine, is still largely unknown. In the present study, a fusion protein (Abeta-EGFP) that combined enhanced green fluorescent protein with amyloid-beta protein (Abeta) was orally administered to mice before and after weaning, and the uptake and kinetics of amyloid protein within the intestine were elucidated through histopathology. Abeta-EGFP was incorporated into the cytoplasm of columnar epithelial cells, rather than M cells, at 3 h after administration and thereafter. Abeta-EGFP then accumulated in the crypt, Peyer's patch, and even the spleen. However, this uptake was not observed in weaned mice. These results suggest that a specific tolerant mechanism for incorporation of Abeta escaped from the digestion exists during suckling periods. This age-dependent uptake is important for estimating the risk of transmission. PMID- 19130398 TI - Chondrocyte-like apoptosis in temporomandibular joint disc internal derangement as a repair-limiting mechanism. An in vivo study. AB - Temporomandibular joint internal derangement (TMJ ID) is characterised by disc displacement and degenerative tissue changes involving an active cellular response, with cell phenotype transformation from fibroblast-like to fibrochondrocyte and, eventually, to chondrocyte-like, possibly as a response to abnormal loading. However, only small patches of chondral tissue are detected in TMJ discs with ID. We decided to explore the reasons for such incomplete tissue change, postulating an involvement of the apoptosis process. Twenty-one discs removed from 19 patients with TMJ ID were processed for TRAIL and DR5 immunohistochemical localisation, and subjected to the TUNEL assay. Overexpression of DR5 receptor and its ligand (TRAIL) in chondrocyte-like cells suggested activation of programmed cell death, as also demonstrated by TUNEL positive cells. The data suggest a failed adaptive response to disc displacement through chondroid metaplasia. The apoptotic death of chondrocyte-like cells, which is at least partly regulated by TRAIL and its death receptor, appears to underpin the failed disc repair, eventually leading to its perforation. PMID- 19130399 TI - Analysis of pRb, p16INK4A proteins and proliferating antigens: PCNA, Ki-67 and MCM5 expression in aggressive fibromatosis (desmoid tumor). AB - Aggressive fibromatosis (desmoid tumor) is a mesenchymal lesion originating from fascial, aponeurotic and muscular connective tissue. It rarely becomes histologically malignant. In this study we analyzed the cell cycle regulation proteins: pRb, p16, and proliferating antigens: Ki-67, PCNA, MCM5 with immunohistochemical method in archival material derived from 27 extra-abdominal (E-AD), 18 abdominal (AD) and 5 intra-abdominal (I-AD) cases of desmoid tumor. None of the examined cases (n=50) of aggressive fibromatosis was pRb immunonegative. Heterogeneous expression of pRb was observed in 51.85% (14/27) of Group AD cases and in 5.56% (1/18) of Group E-AD cases; positive expression in 48,15% (13/27) of Group AD cases, in 94.44% (17/18) of Group E-AD cases, and in 100% (5/5) of Group I-AD cases. There were no negative cases for p16 staining in any of the examined groups. The number of heterogeneous cases in individual groups was: 33.33% (9/27) in Group AD, 50% (9/18) in Group E-AD and 40% (2/5) in Group I-AD, and positive cases: 66.67% (18/27), 50% (9/18) and 60% (3/5), respectively. Overexpression of PCNA was noted in 98% (49/50) of cases. The positive staining for Ki-67 protein was noted in 25.93% (7/27) in Group AD, in 16.67% (3/18) in Group E-AD and in 60% (3/5) in Group I-AD. None of the examined cases was immunopositive for MCM5 protein. The noted levels of pRb and p16 expression in desmoid cells reflect their function in cell cycle regulation. Probably the unsettled cell cycle progression, especially in G1 phase, is not the cause of aggressive fibromatosis pathogenesis. PMID- 19130400 TI - Enhanced CD24 expression in endometrial carcinoma and its expression pattern in normal and hyperplastic endometrium. AB - CD24 is known to be an important diagnostic and prognostic marker of several major cancers affecting females. We aimed to determine CD24 expression in normal, hyperplastic, and carcinomatous endometrium and its correlation with estrogen and progesterone receptor expression. A total of 271 cases including 62 normal/atrophic endometrium cases (47/15), 127 endometrial hyperplasia cases (51/52/24, simple/complex/atypical hyperplasia), and 82 endometrial carcinoma cases were immunohistochemically analyzed by using anti-CD24, ER, and PR antibodies that were embedded on paraffin blocks. Next, we assessed the CD24 mRNA expression in these tissues by using RT-PCR. In the normal endometrium, cyclic expression of membranous CD24 was detected during the regular menstrual cycle, i.e., down-regulation in the proliferative phase and up-regulation in the secretory phase. CD24 expression was very infrequent and weak in the atrophic endometrium. In hyperplasias and carcinomas, the expression of both membranous and cytoplasmic CD24 was found to be sharply reduced in the hyperplastic lesions and significantly enhanced in the carcinomas. In the case of carcinomas, high CD24 expression showed significant correlation with high-grade (G2 and 3) (P<0.05). In addition, an inverse correlation was apparent between CD24 and the estrogen and progesterone receptor expressions in normal and diseased endometrium. In conclusion, we demonstrated that CD24 was expressed in a cyclic pattern in the normal endometrium, and its expression was enhanced in case of endometrial carcinoma. These results suggest that CD24 may be involved in tumor progression and can be a useful diagnostic biomarker. PMID- 19130401 TI - Time-course changes in neural cell apoptosis in the rat fetal brain from dams treated with 6-mercaptopurine (6-MP). AB - 6-Mercaptopurine (6-MP), one of the major drugs for the therapy of acute lymphoblastic leukemia and autoimmune diseases, is incorporated as thioguanine in nucleic acid and it induces cytotoxicity and fetotoxicity. In the present study, pregnant rats were treated with 50 mg/kg of 6-MP on 13 embryonic days (E), and fetuses were collected from 12 to 96 h after the treatment to examine the mechanism and time-course changes in neural cell death in the developing brain. The weights of fetal telencephalon and the thickness of the dorsal telencephalic wall of the fetuses were significantly reduced at 96 h. The number of pyknotic neural cells in the fetal telencephalon began to increase at 24 h, peaked at 36 h, and then gradually decreased toward 72 h. The nuclei of most of these pyknotic cells were stained positively by TUNEL method, which detects DNA fragmentation. Moreover, pyknotic cells were immunohistochemically positive for cleaved caspase 3, one of the key executioners of apoptosis, and the increased expression of the protein from 30 to 48 h was confirmed by using Western blot analysis. Also, electron microscopical features of the pyknotic cells showed ultrastructural characteristics of apoptosis. On the other hand, the number of mitotic and BrdU positive neural cells in the telencephalon decreased from 30 to 72 h. These results suggest that 6-MP induced apoptotic cell death in neural cells in the rat fetal brain is probably due to cytotoxic action of 6-MP. PMID- 19130402 TI - Pulmonary expression of vascular endothelial growth factor (VEGF) and alveolar septation in a newborn rat model exposed to acute hypoxia and recovered under conditions of air or hyperoxia. AB - Vascular endothelial growth factor (VEGF) is an endothelial cell growth factor expressed in normal lung tissue. The aim of the study was to investigate the expression of VEGF and its repercussions as regards alveolarization in the developing rat lung. We studied pulmonary VEGF expression at 0 and 14 days of life in Wistar rats. Rat pups were exposed to hypoxia for two hours during the first hours of life and recovered under conditions of hyperoxia or normoxia for a further two hours, or not recovered. The animals of the control group were only exposed to conditions of normoxia. Our results showed that VEGF was increased in the lungs of the animals that were exposed to hypoxia but we did not find any correlation with the septation. The VEGF was decreased in the lungs of animals exposed to hyperoxia after neonatal hypoxia. We observed this at 0 and 14 days of life, and it was correlated with a lower degree of alveolarization at 14 days of life. Our data suggest that hyperoxia after neonatal hypoxia at birth may give rise to a decrease in the expression of VEGF, possibly permanently, together with a reduction in alveolar development. PMID- 19130403 TI - Langerhans cells in lichen sclerosus of the vulva and lichen sclerosus evolving in vulvar squamous cell carcinoma. AB - Vulvar lichen sclerosus (LS) represents a benign chronic inflammatory skin lesion that carries a risk for development of vulvar squamous cell carcinoma (SCC). We aimed at determining whether premalignant changes in vulvar LS, a multifactorial disease, presenting a welter of evidence implicating the immune system in its pathogenesis, could be identified by analysing the Langerhans' cells (LCs), the primary cell responsible for antigen recognition and presentation. The relationship existing between inflammation and cancer due to chronic infection, and demonstrated in many solid tumors, led us to study LCs in eight cases of vulvar LS, which showed an evolution to carcinoma of the vulva and in ten cases of unchanged vulvar LS in matched patients by immunohistochemistry for antibodies CD1a and S100. We did not find a statistically significantly different number of LCs counted either in S100 stained specimens, nor in CD1a stained specimens of LS epithelium in unchanged or evolving cases. The data emerging in our study do not support the hypothesis that the variation in the number of LCs may be related to the development of SCC in late stage LS cases. PMID- 19130404 TI - Canonical and non-canonical pathways of osteoclast formation. AB - Physiological and pathological bone resorption is mediated by osteoclasts, multinucleated cells which are formed by the fusion of monocyte / macrophage precursors. The canonical pathway of osteoclast formation requires the presence of the receptor activator for NFkappaB ligand (RANKL) and macrophage colony stimulating factor (M-CSF). Non-canonical pathways of osteoclast formation have been described in which cytokines / growth factors can substitute for RANKL or M CSF to induce osteoclast formation. Substitutes for RANKL include LIGHT, TNFalpha and interleukins 6, 11 and 8. M-CSF substitutes include vascular endothelial growth factor (VEGF), placental growth factor (PlGF), FLt-3 ligand and hepatocyte growth factor (HGF). These growth factors can also influence canonical (RANKL / M CSF-induced) osteoclast formation. Both canonical and non-canonical pathways of osteoclast formation play a role in the formation of osteolytic lesions where there is increased osteoclast formation and activity, such as in giant cell tumour of bone. PMID- 19130405 TI - Mesenchymal stem cells in connective tissue engineering and regenerative medicine: applications in cartilage repair and osteoarthritis therapy. AB - Defects of load-bearing connective tissues such as articular cartilage, often result from trauma, degenerative or age-related disease. Osteoarthritis (OA) presents a major clinical challenge to clinicians due to the limited inherent repair capacity of articular cartilage. Articular cartilage defects are increasingly common among the elderly population causing pain, reduced joint function and significant disability among affected patients. The poor capacity for self-repair of chondral defects has resulted in the development of a large variety of treatment approaches including Autologous Chondrocyte Transplantation (ACT), microfracture and mosaicplasty methods. In ACT, a cartilage biopsy is taken from the patient and articular chondrocytes are isolated. The cells are then expanded after several passages in vitro and used to fill the cartilage defect. Since its introduction, ACT has become a widely applied surgical method with good to excellent clinical outcomes. More recently, classical ACT has been combined with tissue engineering and implantable scaffolds for improved results. However, there are still major problems associated with the ACT technique which relate mainly to chondrocyte de-differentiation during the expansion phase in monolayer culture and the poor integration of the implants into the surrounding cartilage tissue. Novel approaches using mesenchymal stem cells (MSCs) as an alternative cell source to patient derived chondrocytes are currently on trial. MSCs have shown significant potential for chondrogenesis in animal models. This review article discusses the potential of MSCs in tissue engineering and regenerative medicine and highlights their potential for cartilage repair and cell-based therapies for osteoarthritis and a range of related osteoarticular disorders. PMID- 19130406 TI - An overview of the pale and clear cells of the nipple epidermis. AB - The stratified squamous epithelium of the nipple-areola complex may contain pale or clear cells including: Paget's disease cells (PDCs), Toker cells (TCs), and so called clear cells (CCs). Paget's disease is an uncommon presentation of breast carcinoma. PDCs are large, atypical, have abundant, pale-staining cytoplasm that may contain mucin secretion vacuoles and bulky heterochromatic nuclei. They are commonly concentrated along the basal layer and stain for EMA, CAM5.2, cytokeratin 7, and HER2/neu oncoprotein. TCs are bland cells with roundish and scant chromatin nuclei. They are found incidentally and are reactive for EMA, CAM5.2, and cytokeratin 7, but show negativity for HER2/neu oncoprotein. So called CCs show varied morphology, are found incidentally, and have been variably interpreted by different authors. The majority of cells that have been called epidermal CCs fit the features of pagetoid dyskeratosis. These cells are reactive for high molecular weight cytokeratin. Other CCs showing signet-ring morphology present negativity for mucins and correspond to a fixation artefact. PMID- 19130407 TI - Lymph node lymphangiogenesis: a new concept for modulating tumor metastasis and inflammatory process. AB - The proliferation of lymphatic endothelial cells (LECs) occurs not only in tumor and inflamed tissues, but also in regional draining lymph nodes (LNs). The lymph node lymphangiogenesis (LNLG) has recently emerged as a prominent area in biomedical research, because it is involved in the pathogenesis of several human diseases. The LEC functional features and lymphatic remodeling regulated by lymphangiogenic factors actively promote tumor metastasis and the inflammation process. VEGF-A/VEGFR-2 and VEGF-C/-D/VEGFR-3 have been implicated as the prime mediators in inflammation- or tumor-induced LNLG. This knowledge may provide a foundation for further understanding of specific modification in the gene expression, cell migration, and differentiation of LECs and other cells in lymphatic-associated diseases. Importantly, it should be taken into consideration that inflammation and lymphangiogenesis are strongly linked in the formation and metastasis of cancer when designing therapeutic strategies. PMID- 19130408 TI - Dysregulation of Hedgehog, Wnt and Notch signalling pathways in breast cancer. AB - There has been a significant decrease in mortality from breast cancer in the last two decades. This has been attributed to the introduction of mammographic screening and to the development of specialised therapies, notably anti-estrogens such as tamoxifen in estrogen receptor (ER) positive tumours, and adjuvant chemotherapy. More recently monoclonal antibodies such as trastuzumab directed against Her2-overexpressing tumours show significant promise in improving outcome from this aggressive subtype. While there have been significant advances, a number of clinical challenges still remain, particularly development of targeted therapies for other forms of breast cancer lacking ER or Her2, such as the aggressive basal-like carcinomas. Identification of new therapeutic targets in poor prognosis groups will be critical to further improvements in breast cancer treatment. Proper functioning of the Hedgehog, Notch and Wnt signalling pathways is required for normal development during early life and these pathways also play a key role in regulation and maintenance of stem cells. Increasing evidence implicates dysregulation of these pathways in the development and progression of a number of malignancies, including breast cancer. This review presents the current evidence for aberrations in these pathways in breast cancer and proposes that the Hedgehog, Notch and Wnt signalling pathways may represent novel therapeutic targets. PMID- 19130409 TI - Natriuretic peptides in heart failure: their role in daily practice. PMID- 19130410 TI - Cardiac resynchronization therapy and atrial fibrillation. AB - Cardiac resynchronization therapy (CRT) is an important advance for the treatment of end-stage heart failure (HF). About 15-50% of HF is complicated by atrial fibrillation (AF), associated with worsened outcomes. The presence of AF may interfere with optimal delivery of CRT due to competition with biventricular (BiV) capture by conducted beats. Pacing algorithms in newer devices may not ensure consistent CRT delivery during periods of rapid ventricular rates. Atrioventricular junction ablation with permanent pacing eliminates interference by conducted beats and provides complete BiV capture and is associated with improved outcomes. Catheter ablation of AF is another promising alternative to maintain sinus rhythm in patients with AF and HF. However, the optimal indications for CRT delivery for patients in this complex cohort remain to be assessed in randomized clinical trials. PMID- 19130411 TI - Bioactivation of organic nitrates and the mechanism of nitrate tolerance. AB - Organic nitrates, such as nitroglycerin, are commonly used in the therapy of cardiovascular disease. Long-term therapy with these drugs, however, results in the rapid development of nitrate tolerance, limiting their hemodynamic and anti ischemic efficacy. In addition, nitrate tolerance is associated with the expression of potentially deleterious modifications such as increased oxidative stress, endothelial dysfunction, and sympathetic activation. In this review we discuss current concepts regarding the mechanisms of organic nitrate bioactivation, nitrate tolerance, and nitrate-mediated oxidative stress and endothelial dysfunction. We also examine how hydralazine may prevent nitrate tolerance and related endothelial dysfunction. PMID- 19130412 TI - Utility of the dobutamine stress echocardiography in the evaluation of the effects of a surgical repair of aortic coarctation in children. AB - BACKGROUND: Exercise-induced hypertension following repair of the coarctation of the aorta (CoA) is a well known phenomenon. The most important functional parameters in the assessment of the effects of a surgical repair of CoA are the maximal pressure gradient in the descending aorta (GRAD) and systolic blood pressure (SBP). Results of treadmill exercise test using the Bruce protocol (treadmill test) and dobutamine stress echocardiography (DSE) were compared to determine utility of the DSE in the evaluation of the effects of surgical treatment of CoA in children. METHODS: The study population comprised of 29 patients, including 20 males and 9 females (mean age 12 years) who underwent a surgical repair of CoA. Changes of the cardiovascular parameters including SBP, GRAD and heart rate (HR) during the treadmill test and DSE were compared. RESULTS: During the treadmill test, SBP at peak exercise ranged from 120 to 230 (mean 163.7) mm Hg, GRAD ranged from 29 to 109 (mean 59.8) mm Hg, and HR ranged from 140 to 188 (mean 169) bpm. At the end of DSE, SBP ranged from 123 to 215 (mean 164.7) mm Hg, GRAD ranged from 29 to 113 (mean 55.4) mm Hg, and HR ranged from 76 to 155 (mean 111) bpm. We found positive correlations of SBP (r = 0.68, p < 0.001) and GRAD (r = 0.82, p < 0.001) values during both tests but no significant correlation for HR (r = 0.42, p = NS). CONCLUSIONS: Dobutamine stress echocardiography is useful in the evaluation of the effects of surgical repair of CoA in children. PMID- 19130413 TI - Imaging morphology of cardiac tumours. AB - BACKGROUND: Cardiac tumours are very uncommon and are the topic of little investigation. Imaging features offer reliable diagnostic evidence for cardiac tumours, but diagnostic confusion may arise when tumours with similar features are present. METHODS: Between January 2003 and July 2008, 34 patients were operated on for cardiac tumours in this institute. The patients' ages ranged from 31 to 81 years with an average of 54.8 +/- 14.2 years. Thirty (88.2%) tumours were primary [19 (55.9%) myxomas, 8 (23.5%) papillary fibroelastomas, and 1 (2.9%) cavernous hemangioma were benign, 1 (2.9%) recurrent fibrous histiocytoma (undifferentiated sarcoma) and 1 (2.9%) leiomyosarcoma were malignant], and 4 (11.8%) were secondary [1 (2.9%) metastatic cardiac leiomyoma, and 3 (8.8%) were renal cell carcinomas]. RESULTS: Cardiac myxomas represented more than half of the cardiac tumours of this patient series, necessitating surgical resection. More than half of these cardiac myxomas originated from the intraatrial septum with a stalk. Most of them appeared as a round or ovoid soft mass on echo, as a hypoattenuated lesion on computed tomography or magnetic resonance imaging, and with a soft gelatinous appearance on gross appearance. Cardiac papillary fibroelastomas were valvular or subvalvular, mostly pedicled by a short stalk, and all of them were pound 1 cm in size. The cavernous hemangioma was isointense on magnetic resonance imaging and tensile and slithy in gross specimen. Recurrent fibrous histiocytoma, leiomyosarcoma, intravenous leiomyoma and renal cell carcinoma resembled a myxoma on echocardiography due to their soft, friable, and mobile features. There were no misdiagnoses based on preoperative imaging features comparable to surgical and histopathologic findings in this surgical series. CONCLUSIONS: Imaging morphology plays a key role in the preoperative differential diagnosis of cardiac tumours. Imaging features could reliably predict primary versus secondary, and benign versus malignant among cardiac tumours. The accurate preoperative imaging assessment of cardiac tumours necessitating surgical resection has become increasingly important in the decision-making of a surgical approach, method, and resection extent. PMID- 19130414 TI - Factors associated with C-reactive protein at the early stage of acute myocardial infarction in men. AB - BACKGROUND: Elevation of C-reactive protein (CRP) is associated with acute coronary events. CRP is related to cardiovascular risk factors and adipokines. The aim of the study was to reveal the factors associated with elevated CRP levels in patients with ST-segment elevation acute myocardial infarction (STEMI). As there are sex-related differences in plasma levels of CRP and adipokines, our study was designed for males. METHODS: Seventy men admitted within the initial 6 hours of STEMI were categorized into 4 groups according to the quartile of CRP. Clinical data and laboratory measurements were analyzed. RESULTS: Anthropometric measurements, glucose at admission, resistin, and leptin were significantly higher, and adiponectin lower with the increase of CRP quartile. A significant positive correlation between CRP and body mass index, waist circumference, glucose at admission, resistin, and leptin and a negative relation of CRP to HDL cholesterol and adiponectin were observed. In univariate logistic regression analysis, variables associated with a level of CRP above the fourth quartile were history of angina, obesity, diabetes, glucose at admission, resistin, leptin, and adiponectin, and independent predictors were glucose at admission and resistin. To predict the elevated CRP level the optimal cut-off for glucose at admission was 144 mg/dL (sensitivity 84%, specificity 86%) and for resistin was 21.5 ng/mL (sensitivity 79%, specificity 71%). CONCLUSIONS: Glucose at admission and resistin are independently associated with elevated levels of CRP in men during the early stage of STEMI. PMID- 19130415 TI - Cutoff values of NT-proBNP for the prediction of low functional capacity, decreased ejection fraction and cardiovascular events in patients with heart failure. AB - BACKGROUND: It has been demonstrated in numerous studies that N-terminal pro-B type natriuretic peptide (NT-proBNP) is strongly associated with left ventricular ejection fraction (LVEF), functional capacity (FC), and cardiovascular (CV) mortality in heart failure (HF) patients. The aim of the present study was to determine the predictive cutoff values of NT-proBNP for predicting these parameters. METHODS: One hundred HF patients (88 male, 12 female, mean age 53.6 +/- 8.9 years) with left ventricular (LV) systolic dysfunction and impaired exercise capacity were enrolled into the study. Echocardiographic examination was performed. The NT-proBNP concentration was measured after resting for 20 min in the supine position. The modified Bruce protocol was utilized for exercise testing. The patients were followed for between 690 and 840 days (mean 750 +/- 30 days) for the occurrence of CV events. RESULTS: There was a strong negative correlation between NT-proBNP concentration and LVEF (p < 0.004). It was found that NT-proBNP is a strong predictor of LVEF < 30% (p < 0.001). When 940 pg/mL was accepted as a cutoff value for NT-proBNP for the prediction of an LVEF < 30%, the sensitivity and the specificity were 89.8% and 71.4%, respectively. NT-proBNP and left atrial diastolic dimension were the most significant parameters for predicting FC (p < 0.001, each one). An NT-proBNP cutoff value of 940 pg/mL responded to 78.8% sensitivity and 81% specificity for the prediction of FC < 5 METs. The observed independent predictors for the CV events were NT-proBNP, LV mass index, and resting heart rate (p < 0.001, p = 0.02 and p = 0.006, respectively). Every 1000 pg/mL elevation in NT-proBNP level resulted in a 27% increase in the occurrence of CV events (p < 0.006). Moreover, 940 pg/mL NT proBNP cutoff value revealed a sensitivity and specificity of 86.7% and 64.7% respectively for the prediction of incident CV events. CONCLUSIONS: Use of NT proBNP cutoff values is easy and reliable method for the prediction of low FC and decreased LVEF, and may aid identification of patients at the highest risk for future CV events. We suggest to use NT-proBNP cutoff value of 940 pg/mL for predicting these parameters. PMID- 19130416 TI - Repolarization parameters in patients with acute ST segment elevation myocardial infarction treated with primary percutaneous coronary intervention with respect to predischarge ST-T pattern: a preliminary study. AB - BACKGROUND: Negative T wave and lack of ST segment elevation in predischarge ECG in ST-segment elevation myocardial infraction (STEMI) patients are given as markers of good prognosis. Repolarization duration, especially its late part (TpeakTend - TpTe), likewise ST-T patterns, is related to local post-myocardial infarction myocardial attributes. We analyzed the differences in QT parameters in STEMI patients with negative or not-negative T wave pattern in predischarge ECG. METHODS: The data from 83 STEMI patients (LVEF > 45%, first MI, one-vessel disease) who underwent successful percutaneous coronary intervention of infarct related coronary artery (TIMI 3 flow) were collected. According to ST-T patterns in predischarge ECG, the cohort was divided into two groups: 38 patients with persistent ST elevation and/or non-negative T wave pattern (STT+), and 45 patients with negative T wave, without ST elevation (STT-). QT, QTpeak, and TpTe intervals were obtained from 5 consecutive beats of sinus rhythm 60-70 bpm between 6 a.m. and 8 a.m. from Holter recording, corrected to the heart rate (HR) with Bazett's formula. RESULTS: The study groups did not differ in gender, age, or treatment. No true antiarrhythmics were given. Both QTc and TpTec were longer in STT+ patients: 459 +/- 26 ms vs. 440 +/- 25 ms, p = 0.01 and 108 +/- 10 ms vs. 96 +/- 11 ms, p = 0.000015, respectively. Prolongation of late repolarization was found both in anterior and inferior infarction. CONCLUSIONS: STEMI patients who underwent successful percutaneous coronary intervention of infarct-related coronary arteries and demonstrated persistent ST elevation, without negative T wave at hospital discharge, had a longer repolarization duration, especially the late phase of it. Further studies are necessary to assess the prognostic value of this finding. PMID- 19130417 TI - Level of blood pressure control in a hypertensive population when measurements are performed outside the clinical setting. AB - BACKGROUND: To determine whether the number of optimally controlled hypertensive patients is higher using self-measurement of blood pressure at home and ambulatory monitoring, compared to using conventional blood pressure measurements at the doctor's office. METHOD: An observational, cross-sectional, multicentre, descriptive study of a random sample of 237 primary health care patients, known to be hypertensive, from Badajoz (Spain). Blood pressure was measured at the doctor's office and by self-measurement at home. Those patients showing good control by self-measurement were subjected to 24-hour ambulatory monitoring. Optimal control was understood as blood pressure < 140/90 mm Hg when measured at the doctor's office, and < 135/85 mm Hg when self-measured at home and by daytime ambulatory monitoring. RESULTS: Mean systolic/diastolic measurements at the doctor's office and by self-measurement were 145.6/83.9 and 134.0/78.7 mm Hg, respectively (p < 0.000). In the population optimally controlled by self measurement and who subsequently received ambulatory monitoring, the mean blood pressure was 121.8/73.4 and 125.6/76.2 mm Hg, respectively (p = 0.002; p < 0.000). When measured at the doctor's office blood pressure was controlled in about 29.5% (95% CI 23.7-35.3%) of patients, in 38% when self-measured (95% CI 31.4-44.2%; p < 0.000), and in 24.5% when it was confirmed through ambulatory monitoring (95% CI 15.4-33.6%). Sensitivity and positive predictive values of the office measurements for the detection of patients who were well-controlled by self-measurement were 50% and 64.3%, respectively, and 53.4% and 73.8% as regards ambulatory monitoring. CONCLUSIONS: A higher level of control is achieved with self-measurement at home not confirmed by ambulatory monitoring. Therefore, the white coat effect does not seem to influence the percentage of well-controlled patients detected at the doctor's office. Office blood pressure does not appear to be useful in distinguishing which individual patients are optimally controlled. PMID- 19130418 TI - Sustained benefit of left ventricular remodelling after valve replacement for aortic stenosis. AB - BACKGROUND: Valve replacement for aortic stenosis (AS) determines negative ventricular remodelling. We used cross sectional and Doppler echocardiography to check how rapidly it occurs and to assess if these changes are sustained over time. METHODS: We evaluated in 34 patients subjected to aortic valve replacement for AS morphological and functional (ejection fraction and E:A ratio) left ventricular data by echocardiography prior to surgery and 2 postoperative studies: early after surgery (pQ1) and at mid-term evolution (pQ2). RESULTS: Left ventricular mass index was reduced at pQ1 (from 152 +/- 47 g/m2 to 113 +/- 31 g/m2; p < 0.01) as well as end-diastolic (from 51.3 mm to 48.3 mm; p < 0.03), end systolic (from 32.2 mm to 29.4 mm; p < 0.02), interventricular septum (from 12.9 mm to 10.3 mm; p < 0.01), and posterior wall (from 12.5 mm to 11 mm; p < 0.01) dimensions. Left ventricular ejection fraction (from 61.2% to 65.2%; p < 0.04) and E:A ratio (from 0.94 to 0.98; p < 0.01) increased significantly at pQ1. There were no significant differences in measurements between pQ1 and pQ2. CONCLUSIONS: Aortic valve replacement surgery leads to a rapid negative left ventricular remodelling during the first 7 months, including a decrease in myocardial hypertrophy and an improvement in systolic and diastolic function. These beneficial hemodynamic changes are sustained for at least 3 years. PMID- 19130419 TI - Wellens' syndrome: the electrocardiographic finding that is seen as unimportant. AB - Wellens' syndrome is a pattern of electrocardiography T-wave changes associated with critical proximal left anterior descending artery lesion. Patients with Wellens' syndrome are at high risk of the development of extensive myocardial infarction of the anterior wall and death. Thus, it is vital that this finding is recognized promptly. We present a patient with Wellens' syndrome in this article. PMID- 19130420 TI - Left ventricular diverticulum, a rare echocardiographic finding: two adult patients and review of the literature. AB - Congenital diverticulum of the left ventricle is a very rare cardiac abnormality characterized by a local embryological development failure of the ventricular muscle. It may present as an isolated disorder or may be associated with other cardiac abnormalities. Clinically, it has been reported that it can lead to chest pain, arrhythmias, cardiac rupture, and sudden death, although frequently the course is asymptomatic. This article presents two illustrative cases of left ventricular diverticulum. The diagnosis was made by two-dimensional echocardiography and was confirmed by cardiac catheterization. There were no other thoracoabdominal or cardiac abnormalities. The patient was asymptomatic and surgical resection was not indicated. Given the few cases reported in the medical literature, the information available is scarce; hence, physicians should be trained to make the appropriate diagnosis. PMID- 19130421 TI - Metoprolol succinate therapy associated with erythema multiforme. AB - Metoprolol succinate is a widely used medication for the management of hypertension, heart failure, and angina. We report the case of a 36-year-old woman who developed erythema multiforme after administration of a low dose of this drug. She also presented with pruritic burning pain throughout her body accompanied by chills. While erythema multiforme has been reported with carvedilol, this is the first observation of metoprolol succinate causing this and physicians should be aware of this potential, yet rare, side-effect. PMID- 19130422 TI - Impending rupture of a giant ascending aortic aneurysm. PMID- 19130423 TI - Implantable electrocardiographic monitoring devices. PMID- 19130424 TI - Access strategies for peripheral arterial intervention. AB - An operator's ability to determine the optimal vascular access strategy for patients undergoing peripheral endovascular intervention is critical to maximizing procedural safety and success. Individualizing an approach to access requires careful planning, and is contingent upon a solid general knowledge of normal and abnormal vascular anatomy, as well as the particulars of each patient's history, physical examination, and non-invasive test results. An awareness of the technical nuances, relative safety, and indications for obtaining percutaneous arterial access at all potential sites is essential. Available means for approaching lower extremity arterial disease include the retrograde and antegrade common femoral approaches, the contralateral crossover technique, upper extremity approaches from the radial, brachial, or axillary arteries, or occasionally retrograde access via the popliteal, dorsalis pedis, or tibial arteries. These techniques, as well as important considerations for approaching disease of the renal, subclavian, and carotid arteries are reviewed. PMID- 19130425 TI - John H. Gibbon Jr., MD: a poet with an idea (1903-1973). PMID- 19130427 TI - Trends in Nanotechnology 2008. PMID- 19130428 TI - Dip-pen-nanolithographic patterning of metallic, semiconductor, and metal oxide nanostructures on surfaces. AB - Dip-pen nanolithography (DPN) is a powerful method to pattern nanostructures on surfaces by the controlled delivery of an "ink" coating the tip of an atomic force microscope upon scanning and contacting with surfaces. The growing interest in the use of nanoparticles as structural and functional elements for the fabrication of nanodevices suggests that the DPN-stimulated patterning of nanoparticles on surfaces might be a useful technique to assemble hierarchical architectures of nanoparticles that could pave methodologies for functional nanocircuits or nanodevices. This Review presents different methodologies for the nanolithographic patterning of metallic, semiconductor, and metal oxide nanostructures on surfaces. The mechanisms involved in the formation of the nanostructures are discussed and the effects that control the dimensions of the resulting patterns are reviewed. The possible applications of the nanostructures are also addressed. PMID- 19130429 TI - Temperature-enhanced solvent vapor annealing of a C3 symmetric hexa-peri hexabenzocoronene: controlling the self-assembly from nano- to macroscale. AB - Temperature-enhanced solvent vapor annealing (TESVA) is used to self-assemble functionalized polycyclic aromatic hydrocarbon molecules into ordered macroscopic layers and crystals on solid surfaces. A novel C3 symmetric hexa-peri hexabenzocoronene functionalized with alternating hydrophilic and hydrophobic side chains is used as a model system since its multivalent character can be expected to offer unique self-assembly properties and behavior in different solvents. TESVA promotes the molecule's long-range mobility, as proven by their diffusion on a Si/SiO(x) surface on a scale of hundreds of micrometers. This leads to self-assembly into large, ordered crystals featuring an edge-on columnar type of arrangement, which differs from the morphologies obtained using conventional solution-processing methods such as spin-coating or drop-casting. The temperature modulation in the TESVA makes it possible to achieve an additional control over the role of hydrodynamic forces in the self-assembly at surfaces, leading to a macroscopic self-healing within the adsorbed film notably improved as compared to conventional solvent vapor annealing. This surface re organization can be monitored in real time by optical and atomic force microscopy. PMID- 19130434 TI - Experience with a Punjabi, Urdu and Hindi rheumatology telephone helpline. AB - OBJECTIVES: To investigate what proportion of patients attending a rheumatology unit in Birmingham, UK, require interpretation services and to assess the use of an Asian language telephone helpline we have established for those who find it easier to communicate in Punjabi, Urdu or Hindi than in English. METHODS: Our patients' requirement for interpretation services and their ability to read the script of the language they preferred to communicate in was assessed. A second survey assessed the use of an Asian language helpline we had established. All calls were recorded over 18 months. The reasons for the calls were categorized and compared with those made to our English helpline. RESULTS: 171 of 512 patients (33%) stated that they required interpretation services. 128 (25%) were not able to read the script of their preferred language. In the second survey, 101 calls to the Asian language helpline were assessed and were compared with calls to the English helpline. The reasons for the calls differed between the helplines. CONCLUSIONS: A large proportion of patients attending our rheumatology unit required interpretation services to communicate effectively with their healthcare professional. Most of these patients were unable to read the script of the language they could speak, casting some doubt over the utility of translated written information. Patients of South Asian origin who preferred to communicate in Punjabi, Urdu or Hindi made use of a helpline operating in these languages, suggesting that this may be an effective strategy to facilitate communication with this group of patients. PMID- 19130435 TI - Synthesis of large-pore urea-bridged periodic mesoporous organosilicas. AB - In this article we report the successful synthesis of a new class of periodic mesoporous organosilicas (PMOs) with a urea-bridged organosilica precursor under acid-catalyzed and inorganic-salt-assisted conditions. The large-pore hybrid materials have an ordered mesostructure with uniform pore size distributions, excellent thick framework walls, thermal stability, and specific functional groups. The composition of the mesoporous organosilicas was characterized by FTIR spectroscopy, (13)C cross-polarization magic-angle spinning (CP MAS) NMR spectroscopy, and (29)Si MAS NMR spectroscopy. PMID- 19130436 TI - Bridging M-Cl bonds with ambiphilic phosphine-borane ligands. AB - A combined experimental/theoretical study provides insight into the bridging coordination of M-Cl bonds with the ambiphilic phosphine-borane ligands iPr(2)P o(C(6)H(4))-BR(2) (PBCy(2): R = Cy; PBMes(2): R = Mes). Reaction of [PdCl(allyl)(PBCy(2))] (3) with HCl affords the related dinuclear complex [PdCl(mu-Cl)(PBCy(2))](2) (5). Subsequent cleavage of the chloride bridge by PPh(3) leads to the heteroleptic mononuclear complex trans [PdCl(2)(PPh(3))(PBCy(2))] (6). The solid-state structures of complexes 5 and 6 substantiate the propensity of the PBCy(2) ligand to bridge Pd-Cl bonds via P- >Pd--Cl-->B interactions. DFT calculations carried out on both the model mononuclear complexes 3* and 6* reveal that in each system the energy of the linkage isomer with a Cl-->B interaction is very similar to that without. A comparison of the solution and solid-state (11)B NMR spectroscopic data for complexes 3 and 6 suggests the possible interconversion of the bridging and B pendant forms in solution. Bridging coordination of the PBCy(2) ligand across a Rh-Cl bond is observed in the solid-state structure of the related complex [RhCl(nbd)(PBCy(2))] (7). Replacement of the Cy groups at boron by Mes substituents illustrates the role of steric factors on the participation of the Lewis acid upon coordination, no Cl-->B interaction being observed in the complex [PdCl(allyl)(PBMes(2))] (8). PMID- 19130437 TI - Unusual domino michael/aldol condensation reactions employing oximes as N selective nucleophiles: synthesis of N-hydroxypyrroles. PMID- 19130438 TI - The copper-catalyzed decarboxylative coupling of the sp3-hybridized carbon atoms of alpha-amino acids. PMID- 19130446 TI - Nano-supramolecular assemblies constructed from water-soluble bis(calix[5]arenes) with porphyrins and their photoinduced electron transfer properties. AB - Possessing 2D netlike and 1D linear morphologies, two nano-supramolecular architectures A1 and A2 are constructed by tetracationic porphyrin (G1) and dicationic porphyrin (G2), respectively, upon complexation with the novel water soluble bis(p-sulfonatocalix[5]arenes) bridged at the lower rim (H2). Corresponding to the molecular design, the aggregation morphologies are well manipulated by the inherent binding sites of the building blocks through host guest interactions as well as charge interactions. In comparison to the simple p sulfonatocalix[5]arene H1 which can only form particle-type complexes C1 and C2 with porphyrin guests, H2 provides excellent pre-organized structure to construct highly complex nano-supramolecular assemblies. The exhibited electron-transfer process of the supramolecular systems is further investigated by steady-state and time-resolved fluorescence spectroscopy, electrochemical measurements, and transient absorption spectroscopy. The results obtained show that calixarenes are also effective electron donors in PET besides acting as significant building blocks, which gives them many advantages in constructing well-ordered nanomaterials with the capability of electron and energy transport. PMID- 19130447 TI - Dithiolate-bridged Fe-Ni-Fe trinuclear complexes consisting of Fe(CO)(3-n)(CN)(n) (n = 0, 1) components relevant to the active site of [NiFe] hydrogenase. AB - A dithiolate-bridged Fe-Ni-Fe trinuclear carbonyl complex [(CO)(3)Fe(mu-ndt)Ni(mu ndt)Fe(CO)(3)] (1, ndt = norbornane-exo-2,3-dithiolate) has been synthesized from the reaction of [Fe(CO)(4)I(2)] and Li(2)[Ni(ndt)(2)]. This reaction was found to occur with concomitant formation of a tetranuclear cluster [Ni(3)(mu-ndt)(4)FeI] (2). Treatment of 1 with Na[N(SiMe(3))(2)] transforms some of the CO ligands into CN(-), and the monocyanide complex (PPh(4))[(CO)(2)(CN)Fe(mu-ndt)Ni(mu ndt)Fe(CO)(3)] (3) and the dicyanide complex (PPh(4))(2)[(CO)(2)(CN)Fe(mu ndt)Ni(mu-ndt)Fe(CO)(2)(CN)] (4) were isolated. X-ray structural analyses of the trinuclear complexes revealed a Fe-Ni-Fe array in which the metal centers are connected by the ndt sulfur bridges and direct Fe-Ni bonds. Hydrogen bonding between the CN ligand in 3 and cocrystallized ethanol was found in the solid state structure. The monocyanide complex 3 and dicyanide complex 4 reacted with acids such as HOTf or HCl generating insoluble materials, whereas complex 1 did not react. PMID- 19130451 TI - Guest Editorial: from Chem. Eur. J. to ChemSusChem: all from ChemPubSoc Europe. PMID- 19130452 TI - Discrete supramolecular donor-acceptor complexes. PMID- 19130454 TI - Capsaicin: tailored chemical defence against unwanted "frugivores". AB - Why are chilli peppers hot? The vanillyl amide, capsaicin, has long been known as the pungent principle of peppers, but only in their recent work have Tewksbury et al. addressed its ecological roles: to distract unsuitable seed dispensers and to protect the seeds against fungal infection by Fusarium. PMID- 19130455 TI - Formylglycine aldehyde Tag--protein engineering through a novel post translational modification. AB - Oxidation of a specific cysteine residue to C(alpha)-formylglycine is a novel post-translational modification that is directed by a short recognition motif commonly found in pro- and eukaryotic sulfatases. As recently shown by C. Bertozzi and co-workers, this system can be employed in protein engineering to equip proteins with genetically encoded aldehyde tags for site-specific labeling, conjugation and immobilization. PMID- 19130444 TI - Recent advances in the chemistry and biology of naturally occurring antibiotics. AB - Ever since the world-shaping discovery of penicillin, nature's molecular diversity has been extensively screened for new medications and lead compounds in drug discovery. The search for agents intended to combat infectious diseases has been of particular interest and has enjoyed a high degree of success. Indeed, the history of antibiotics is marked with impressive discoveries and drug-development stories, the overwhelming majority of which have their origin in natural products. Chemistry, and in particular chemical synthesis, has played a major role in bringing naturally occurring antibiotics and their derivatives to the clinic, and no doubt these disciplines will continue to be key enabling technologies. In this review article, we highlight a number of recent discoveries and advances in the chemistry, biology, and medicine of naturally occurring antibiotics, with particular emphasis on total synthesis, analogue design, and biological evaluation of molecules with novel mechanisms of action. PMID- 19130456 TI - Triclosan derivatives: towards potent inhibitors of drug-sensitive and drug resistant Mycobacterium tuberculosis. AB - Triclosan has been previously shown to inhibit InhA, an essential enoyl acyl carrier protein reductase involved in mycolic acid biosynthesis, the inhibition of which leads to the lysis of Mycobacterium tuberculosis. Using a structure based drug design approach, a series of 5-substituted triclosan derivatives was developed. Two groups of derivatives with alkyl and aryl substituents, respectively, were identified with dramatically enhanced potency against purified InhA. The most efficacious inhibitor displayed an IC(50) value of 21 nM, which was 50-fold more potent than triclosan. X-ray crystal structures of InhA in complex with four triclosan derivatives revealed the structural basis for the inhibitory activity. Six selected triclosan derivatives were tested against isoniazid-sensitive and resistant strains of M. tuberculosis. Among those, the best inhibitor had an MIC value of 4.7 microg mL(-1) (13 microM), which represents a tenfold improvement over the bacteriocidal activity of triclosan. A subset of these triclosan analogues was more potent than isoniazid against two isoniazid-resistant M. tuberculosis strains, demonstrating the significant potential for structure-based design in the development of next generation antitubercular drugs. PMID- 19130457 TI - Outcomes of stereotactic radiotherapy for a new clinical stage I lung cancer arising postpneumonectomy. AB - BACKGROUND: Resection of second primary lung tumors that arise after previous pneumonectomy is associated with a high risk of complications. In this study, the authors reviewed outcomes after stereotactic radiation therapy (SRT) for such patients. METHODS: SRT was undergone by 15 patients who developed a new clinical stage I lung cancer at a median of 8.9 years postpneumonectomy, half of whom had severe chronic obstructive pulmonary disorder (COPD). SRT target volumes encompassed all respiratory motion using 4-dimensional computed tomography (CT) scans, and risk-adapted radiation schemes that ranged from 3 x 20 grays (Gy) to 8 x 7.5 Gy were used, depending on tumor size, location, and overlap with prior radiation treatment. All schemes had a biologic effective dose >100 Gy. Follow-up CT scans were obtained at 3 months, 6 months, and 12 months after SRT and yearly thereafter. RESULTS: At a median follow-up of 16.5 months, no local failures were observed, and only 2 patients experienced grade > or = 3 toxicity. One patient had transient pneumonitis that required steroids, and another patient required an increase in oxygen use. The 1-year actuarial disease-free survival rate was 92%. One patient died 10 months post-SRT after developing regional and distant metastases, and 1 patient developed an isolated regional failure. All other patients remained alive and disease free. CONCLUSIONS: SRT was a safe and effective treatment for stage I lung tumors that arose after prior pneumonectomy, even in patients who had severe COPD. SRT was tolerated well, and the current findings suggest that surveillance for second tumors is indicated in all patients after pneumonectomy. PMID- 19130458 TI - Phase II trial of neoadjuvant docetaxel and gefitinib followed by radical prostatectomy in patients with high-risk, locally advanced prostate cancer. AB - BACKGROUND: Prostate cancer trials investigating neoadjuvant hormonal therapy, followed by surgery, have demonstrated that elimination of all tumor cells from the primary site is rare. The authors report a phase 2 trial assessing the efficacy and toxicity of docetaxel and gefitinib in patients with high-risk localized prostate cancer as neoadjuvant therapy before radical prostatectomy (RP). METHODS: Thirty-one patients with high-risk prostate cancer were treated with docetaxel and gefitinib for 2 months before RP. All patients met the criteria of clinical stage T2b-3 or serum prostate-specific antigen (PSA) level >20 ng/mL, or Gleason score of 8 to 10. The primary endpoint was pathologic complete response. Secondary objectives included clinical response. When available, endorectal coil magnetic resonance imaging (eMRI) was performed as part of clinical response evaluation. Immunohistochemical staining of epidermal growth factor receptor and HER-2/neu was performed on prechemotherapy and postchemotherapy prostate tissue. RESULTS: The median age of the patients was 60 years, the median pretreatment PSA level was 7.43 ng/mL, and the median Gleason score was 8. Clinical staging prior to treatment consisted of: T1 in 4 patients, T2 in 17 patients, and T3 in 10 patients. One patient with enlarged pelvic adenopathy and T4 disease did not undergo RP. Thirty patients received all scheduled therapies including RP. Grade 3 toxicities included asymptomatic liver function test elevation in 4 (13%) patients, diarrhea in 1 (3%) patient, and fatigue in 1 (3%) patient. One patient experienced grade 4 toxicity with elevated alanine aminotransferase. RP specimen pathology demonstrated residual carcinoma in all cases. Twenty-nine (94%) patients achieved a clinical partial response, including 35% of patients who demonstrated radiographic improvement on eMRI. CONCLUSIONS: No pathologic complete response was noted in 31 patients treated with docetaxel and gefitinib. This combination was well tolerated, and did not result in increased surgical morbidity. PMID- 19130460 TI - A possible association between aprotinin and improved survival after radical surgery for mesothelioma. AB - BACKGROUND: Aprotinin has been used to decrease blood loss with complicated cardiac surgery but has not been investigated in extrapleural pneumonectomy, an operation that does not use cardiopulmonary bypass. In this prospective, randomized, placebo-controlled, double-blind trial, the authors investigated whether aprotinin decreased blood loss in patients who underwent this operation. METHODS: After appropriate statistical design and institutional review board approval, eligible patients who were scheduled for extrapleural pneumonectomy were randomized to receive either aprotinin or placebo during the operation. Blood loss and survival data were obtained from electronic medical records and surgical databases. RESULTS: Of 20 patients who were enrolled, 16 patients met criteria for blood loss analysis. Four patients were excluded from the blood loss analysis: Three patients were inoperable because of tumor spread and underwent limited surgery, and 1 patient died intraoperatively because of acute, massive hemorrhage. The mean blood loss was 769 mL with aprotinin versus 1832 mL with placebo (P = .05; Wilcoxon test). All 20 patients were included in survival analyses. All 9 patients who received placebo died. In contrast, 7 of 11 patients who received aprotinin remained alive at the time of the current report. Kaplan Meier survival curves differed significantly between the 2 groups (P = .0004). A Bayesian multivariate survival analysis of 18 patients who had complete data available on 8 prognostic variables indicated a posterior probability of .99 that aprotinin was beneficial. CONCLUSIONS: Aprotinin decreased blood loss. After accounting for covariate effects, there was a significant comparative benefit with aprotinin in postoperative survival. This finding was unexpected and could not be considered conclusive because of the small size of the current study. A confirmatory study may be warranted. PMID- 19130459 TI - Fragile histidine triad protein, WW domain-containing oxidoreductase protein Wwox, and activator protein 2gamma expression levels correlate with basal phenotype in breast cancer. AB - BACKGROUND: The expression of fragile histidine triad protein (Fhit) and WW domain-containing oxidoreductase protein (Wwox), tumor suppressors that are encoded by fragile (FRA) loci FRA3B and FRA16D, are lost concordantly in breast cancers. In the current study, the authors examined correlations among Fhit, Wwox, the activator protein 2 transcription factors AP2alpha and AP2gamma, cytokeratins 5 and 6 (CK5/6), epidermal growth factor receptor (EGFR), estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) and their associations with breast cancer phenotypes. METHODS: Tissue microarrays constructed from 837 breast cancer blocks were immunostained. Expression in >10% of tumor cells was considered positive for cytoplasmic CK5/6, membranous EGFR, and nuclear AP2alpha and AP2gamma. Cytoplasmic Fhit and Wwox staining was scored according to staining intensity. ER, PR, and HER-2 status of tumors was derived from records. Correlations among immunohistochemical markers and tumor subtypes were assessed by univariate and multivariate statistical methods. RESULTS: Triple-negative tumors had more frequent expression of EGFR, CK5/6 (P < .001), and AP2gamma (P = .003) and more frequent loss of Fhit and Wwox (P < .001), and an inverse correlation was observed between Fhit, Wwox expression and EGFR, ER, and PR expression (P < .001). Reduced Fhit expression was more common in HER-2-positive and AP2gamma-positive cases (P < .001 and P = .002, respectively). There was a direct correlation noted between Fhit and Wwox (P < .001) and a borderline positive relation between AP2alpha and AP2gamma (P = .054). CONCLUSIONS: The results from this investigation suggested that reduced expression levels of Fhit, Wwox, and nuclear AP2gamma have roles in the pathogenesis of basal-like differentiation in breast cancer. Alteration in the expression of fragile site genes occurs in most of these cancers and may contribute to defects in DNA repair, as observed in breast cancer 1 (BRCA1) deficient cancers. Thus, DNA damage response checkpoint proteins may be targets for treatment. PMID- 19130461 TI - Behavioral, social, and educational outcomes after pediatric stem cell transplantation and related factors. AB - BACKGROUND: The purpose of the current study was to investigate longitudinally children's behavioral and social competence outcomes up to 2 years after pediatric stem cell transplantation (SCT) and related factors. METHODS: Ninety nine mothers and 24 youths completed standardized questionnaires (Child Behavior Checklist [CBCL] and Youth Self-Report [YSR]) pre-SCT, and 12 and 24 months after SCT; 26 teachers completed Teacher Report Form (TRF) at 24 months. Information regarding clinical (eg, diagnosis, cranial radiation, graft vs host disease [GVHD]), child (eg, age, sex, physical health), and familial (eg, maternal age, education, distress) factors was also obtained. RESULTS: Linear mixed regression models with compound covariance structure followed by adjusted pairwise analyses yielded significant improvements from pre-SCT to 1 and 2 years after SCT in total scores; in externalizing and internalizing scores from pre-SCT to 2 years after SCT; and in total competence from pre-SCT to 1 year after SCT. Child's physical health, maternal age, and depression were found to be significantly associated with the total, internalizing, and externalizing CBCL scores, whereas GVHD, mother's age, and time since diagnosis were associated with CBCL competence scores. Diagnosis, cranial radiation, GVHD, child's physical health, and maternal age and education were associated with YSR total behavioral and competence scores. Finally, total TRF scores were associated with time since diagnosis; TRF educational and adaptability scores were associated with maternal education, age, and distress. CONCLUSIONS: Clinical, personal, and familial factors must be considered to understand the psychosocial outcomes of these survivors up to 2 years after SCT. This study has important implications for psychosocial interventions for this population. PMID- 19130462 TI - Survival after hepatic resection of colorectal cancer metastases: a national experience. AB - BACKGROUND: Most estimates of short- and long-term survival after hepatic resection of colorectal cancer metastases are derived from surgical case series. For the current report, the authors used Medicare data to investigate operative mortality and long-term survival in a national sample and examined the factors associated with survival. METHODS: Data were analyzed from Medicare enrollees (age >or=65 years) who were admitted to hospital between January 1, 2000 and December 31, 2004 with a primary diagnosis of colorectal cancer with resection. The sample was restricted to patients who subsequently underwent hepatic resection for liver metastases. The Medicare Denominator File was used to determine operative mortality and long-term survival and the factors that were associated with those outcomes. RESULTS: Of the 306,061 Medicare beneficiaries who were diagnosed with colorectal cancer, 3957 patients were identified who underwent hepatic resection for liver metastases. The crude 30-day and 90-day mortality rates were 4% and 8.2%, respectively, and the 5-year survival rate was 25.5%. Advancing age (hazards ratio [HR], 1.83; 95% confidence interval [95% CI], 1.32-2.53 for age >or=80 years vs ages 65-69 years), comorbid disease (HR, 1.40; 95% CI, 1.06-1.85 for Charlson >or=5 vs Charlson 0), and synchronous colon/hepatic resection (HR, 2.46; 95% CI, 1.89-3.20 for synchronous vs metachronous resection) were associated with worse 90-day mortality. Similarly, long-term mortality was associated with age (HR, 1.36; 95% CI, 1.18-1.56), comorbid disease (HR, 1.51; 95% CI, 1.36-1.69), and synchronous colon/hepatic resection (HR, 1.37; 95% CI, 1.24-1.51 for synchronous vs metachronous resection). CONCLUSIONS: In this national study, short- and long-term survival was worse than that reported in surgical case series. Subgroups at high risk for worse outcomes include the extreme elderly and those undergoing synchronous colon and hepatic resection. PMID- 19130464 TI - Pancreatic neuroendocrine tumors: the impact of surgical resection on survival. AB - BACKGROUND: Although surgical resection is generally recommended for patients with localized pancreatic neuroendocrine tumors (PNETs), the impact of resection on overall survival is unknown. The authors investigated the survival advantage of pancreatic resection using a national database. METHODS: This is a retrospective survival analysis of patients with PNETs from the Surveillance, Epidemiology, and End Results database (1988-2002). RESULTS: A total of 728 patients with PNETs were identified with a median survival of 43 months using Kaplan-Meier survival methods. Resection of tumor was associated with significantly improved survival compared with those patients who were recommended for but did not undergo resection (114 months vs 35 months; P < .0001). This survival benefit was demonstrated for patients with localized, regional, and metastatic disease. A multivariable Cox proportional hazards model was constructed to assess the overall effect of surgical resection on survival, and demonstrated an adjusted odds ratio of 0.48 (95% confidence interval, 0.35-0.66) compared with those who were recommended for surgery but did not proceed to surgery. CONCLUSIONS: The authors have demonstrated in a large national study that resection of primary tumor in patients with PNETs is associated with improved survival across all disease stages. Patients with localized, regional, and metastatic PNETs who are reasonable operative candidates should be considered for resection of their primary tumors. PMID- 19130463 TI - Racial disparities in the development of breast cancer metastases among older women: a multilevel study. AB - BACKGROUND: Distant metastases are the most common and lethal type of breast cancer relapse. The authors examined whether older African American breast cancer survivors were more likely to develop metastases compared with older white women. They also examined the extent to which 6 pathways explained racial disparities in the development of metastases. METHODS: The authors used 1992-1999 Surveillance, Epidemiology, and End Results (SEER) data with 1991-1999 Medicare data. They used Medicare's International Classification of Diseases, Ninth Revision, Clinical Modification codes to identify metastases of respiratory and digestive systems, brain, bone, or other unspecified sites. The 6 pathways consisted of patient characteristics, tumor characteristics, type of treatment received, access to medical care, surveillance mammography use, and area-level characteristics (poverty rate and percentage African American) and were obtained from the SEER or Medicare data. RESULTS: Of the 35,937 women, 10.5% developed metastases. In univariate analysis, African American women were 1.61 times (95% confidence interval [CI], 1.54-1.83) more likely to develop metastasis than white women. In multivariate analysis, tumor grade, stage at diagnosis, and census-tract percentage African American explained why African American women were more likely to develop metastases than white women (hazard ratio, 0.84; 95% CI, 0.68-1.03). CONCLUSIONS: Interventions to reduce late-stage breast cancer among African Americans also may reduce racial disparities in subsequent increased risk of developing metastasis. African Americans diagnosed with high-grade breast cancer could be targeted to reduce their risk of metastasis. Future studies should identify specific reasons why the racial distribution in census tracts was associated with racial disparities in the risk of breast cancer metastases. PMID- 19130465 TI - Cognitive-behavioural therapy for individuals with bulimia nervosa and a co occurring substance use disorder. AB - A significant percentage of individuals with bulimia nervosa (BN) also can be diagnosed with a co-occurring substance use disorder (SUD). Although studies have addressed the frequency of overlap between the disorders, etiology and shared personality traits, limited research is available about the treatment of these comorbid patients. Adapting cognitive-behaviour therapy (CBT) to serve as an integrated treatment for patients with both BN and a SUD is a viable option, as studies of CBT suggest that this form of treatment is efficacious for both disorders independently. The shared strategies in CBT for BN and SUDs facilitate the development of a combined treatment for individuals with both disorders with the addition of modules designed to address some common features of these disorders, such as motivation, difficulty with interpersonal relationships, reward sensitivity and impulsivity. Future research should begin to evaluate the efficacy of an integrated CBT in treating individuals with BN and a SUD. PMID- 19130466 TI - Influence of the baseline 18F-fluoro-2-deoxy-D-glucose positron emission tomography results on survival and pathologic response in patients with gastroesophageal cancer undergoing chemoradiation. AB - BACKGROUND: In patients with esophageal cancer who receive chemoradiation, tools to predict/prognosticate outcome before administering therapy are lacking. The authors evaluated initial standardized unit value (iSUV) of 18F-fluoro-2-deoxy-D glucose positron emission tomography and its association with overall survival and the degree of pathologic response after surgery. METHODS: The authors analyzed 161 patients with esophageal adenocarcinoma who had chemoradiation followed by surgery. The log-rank test, univariate Cox proportional hazards model, Kaplan-Meier survival plot, and Fisher exact test were used to analyze dichotomized iSUV and its association with overall survival and pathologic response. RESULTS: The median age of 161 patients was 61 years (range, 26-80 years) and the majority of patients had lower esophageal or gastroesophageal junction involvement. All patients received fluoropyrimidine and, most commonly, a taxane or platinum compound with concomitant radiation. The median radiation dose was 45 grays (Gy) (range, 45 Gy-50.4 Gy). The median iSUV for all patients was 10.1 (range, 0-58). Using the Fisher exact test, iSUV was not found to be associated with the location of the primary cancer. iSUV higher than the median (10.1) was associated with a better pathologic response (P = .06). Patients with primary cancer with iSUV >10.1 had a lower risk for death (hazards ratio of 0.56) compared with those with iSUV < or = 10.1. Higher iSUV was nonsignificantly associated with improved survival (P = .07). CONCLUSIONS: Data from the current study suggest that lower iSUV is associated with poor survival and lower probability of response to chemoradiation. iSUV needs to be further evaluated because it may be used to complement other imaging or biomarker assessments to individualize therapy. PMID- 19130467 TI - Parental attitudes, body image disturbance and disordered eating amongst adolescents and young adults: a review. AB - OBJECTIVE: The purpose of this paper was to review the existing literature regarding the contribution of parental influences to the sociocultural pressures on body image disturbance and disordered eating so as to highlight principal findings so that parents can be given practical information and identify areas that require further research. METHODS: Relevant articles were located through Pubmed, Sciencedirect and PsychInfo, as well as the screening of bibliographies. RESULTS: The available data suggest that parents are strong communicators of sociocultural pressures. Parental influences via verbal messages and active encouragement have been shown to have more impact on offspring's body concerns and eating behaviours than modelling effects. Both mothers and fathers are important sources of influence for their offspring. CONCLUSION: Considering the role of parents could help improve public health management. Futher exploration of the way adolescents and young adults interpret and perceive parental attitudes and of potential protective factors is necessary. PMID- 19130468 TI - Brain temperature and pH measured by (1)H chemical shift imaging of a thulium agent. AB - Temperature and pH are two of the most important physiological parameters and are believed to be tightly regulated because they are intricately related to energy metabolism in living organisms. Temperature and/or pH data in mammalian brain are scarce, however, mainly because of lack of precise and non-invasive methods. At 11.7 T, we demonstrate that a thulium-based macrocyclic complex infused through the bloodstream can be used to obtain temperature and pH maps of rat brain in vivo by (1)H chemical shift imaging (CSI) of the sensor itself in conjunction with a multi-parametric model that depends on several proton resonances of the sensor. Accuracies of temperature and pH determination with the thulium sensor - which has a predominantly extracellular presence - depend on stable signals during the course of the CSI experiment as well as redundancy for temperature and pH sensitivities contained within the observed signals. The thulium-based method compared well with other methods for temperature ((1)H MRS of N-acetylaspartate and water; copper-constantan thermocouple wire) and pH ((31)P MRS of inorganic phosphate and phosphocreatine) assessment, as established by in vitro and in vivo studies. In vitro studies in phantoms with two compartments of different pH value observed under different ambient temperature conditions generated precise temperature and pH distribution maps. In vivo studies in alpha-chloralose anesthetized and renal-ligated rats revealed temperature (33-34 degrees C) and pH (7.3-7.4) distributions in the cerebral cortex that are in agreement with observations by other methods. These results show that the thulium sensor can be used to measure temperature and pH distributions in rat brain in vivo simultaneously and accurately using Biosensor Imaging of Redundant Deviation in Shifts (BIRDS). PMID- 19130469 TI - Budesonide nanoparticle agglomerates as dry powder aerosols with rapid dissolution. AB - Nanoparticle technology represents an attractive approach for formulating poorly water-soluble pulmonary medicines. Unfortunately, nanoparticle suspensions used in nebulizers or metered dose inhalers often suffer from physical instability in the form of uncontrolled agglomeration or Ostwald ripening. In addition, processing such suspensions into dry powders can yield broad particle size distributions. To address these encumbrances, a controlled nanoparticle flocculation process has been developed. Nanosuspensions of the poorly water soluble drug budesonide were prepared by dissolving the drug in organic solvent containing surfactants followed by rapid solvent extraction in water. Different surfactants were employed to control the size and surface charge of the precipitated nanoparticles. Nanosuspensions were flocculated using leucine and lyophilized. Selected budesonide nanoparticle suspensions exhibited an average particle size ranging from approximately 160 to 230 nm, high yield and high drug content. Flocculated nanosuspensions produced micron-sized agglomerates. Freeze drying the nanoparticle agglomerates yielded dry powders with desirable aerodynamic properties for inhalation therapy. In addition, the dissolution rates of dried nanoparticle agglomerate formulations were significantly faster than that of stock budesonide. The results of this study suggest that nanoparticle agglomerates possess the microstructure desired for lung deposition and the nanostructure to facilitate rapid dissolution of poorly water-soluble drugs. PMID- 19130470 TI - Novel methodology for predicting photogenotoxic risk of pharmaceutical substances based on reactive oxygen species (ROS) and DNA-binding assay. AB - Drug-induced phototoxic skin responses involves photoirritation, photoallergy, and photogenotoxicity. In the present study, we attempted to develop effective analytical tools to predict the photogenotoxic potential of pharmaceutical substances on the basis of their photobiological and DNA-binding properties. Photocleavage of plasmid DNA was assessed by capillary gel electrophoresis (CGE), and generation of reactive oxygen species (ROS) from photoirradiated photosensitizers was also assessed by spectrophotometrical determination. The affinity of drugs with DNA was determined by fluorescent titration or competitive binding assay using ethidium bromide. Structural transition from supercoiled form to open circular form of plasmid DNA were observed in the presence of nine irradiated-photosensitizers, however DNA damage by six phototoxic and five nonphototoxic drugs were negligible or weak even though some chemicals exhibited significant generation of ROS. DNA photocleavers tended to interact with DNA, binding constants of which were found to be less than 1 microM, whereas most nonphotogenotoxic chemicals failed to bind with DNA. The differences in DNA binding properties might be attributed to a data discrepancy between ROS generation and DNA photocleaving activity. The combination use of ROS assay and DNA-binding assay could be fast and reliable prediction for the photogenotoxic potential of a large number of drug candidates at the drug-discovery stage. PMID- 19130471 TI - Spheronization of small extrudates containing kappa-carrageenan. AB - Spheronization of extrudates of around 500 microm diameter needs improvement of the Schlueter spheronizer conditions with regard to moisture content of the extrudates. The extrudates were obtained by a twin-screw extruder and contained kappa-carrageenan as pelletization aid. The influences of spheronization speed, residence time, temperature of the spheronizer wall and loading on the responses aspect ratio, pellet size and yield, were studied with a central composite circumscribed design. The Schlueter spheronizer was compared with a Nica spheronizer. Further, additional spheronizer process variables such as temperature of the spheronizer wall and inlet air pressure were also investigated. The results were evaluated in a full factorial (mixed) design. The micropellets in general showed a pellet size between 500 and 700 microm. A twisted-rope movement during the spheronization process was not observed and adhesion to the spheronizer wall resulted in suboptimal micropellets. However, at suitable moisture content, less loading in the spheronizer, higher spheronization speed and longer residence time micropellets with an aspect ratio below 1.1 were obtained. In addition the adhesion to the spheronizer wall was reduced. Spheronizer wall temperature and inlet air pressure were negligible variables. Significant differences between the two spheronizers could not be established. PMID- 19130472 TI - Detection of protein aggregates by sedimentation velocity analytical ultracentrifugation (SV-AUC): sources of variability and their relative importance. AB - Sedimentation velocity analytical ultracentrifugation (SV-AUC) has found application in the biopharmaceutical industry as a method of detecting and quantifying protein aggregates. While the technique offers several advantages (i.e., matrix-free separation and minimal sample handling), its results exhibit a high degree of variability relative to orthogonal size-sensitive separation techniques such as size exclusion chromatography (SEC). The goal of this work is to characterize and quantify the sources of variability that affect SV-AUC results, particularly size distributions for a monoclonal antibody monomer/dimer system. Contributions of individual factors to the overall variability are examined. Results demonstrate that alignment of sample cells to the center of rotation is the most significant contributing factor to overall variability. The relative importance of other factors (e.g., temperature equilibration, time invariant noise, meniscus misplacement, etc.) are quantified and discussed. PMID- 19130474 TI - The cellular niche of Listeria monocytogenes infection changes rapidly in the spleen. AB - The spleen is an important organ for the host response to systemic bacterial infections. Many cell types and cell surface receptors have been shown to play role in the capture and control of bacteria, yet these are often studied individually and a coherent picture has yet to emerge of how various phagocytes collaborate to control bacterial infection. We analyzed the cellular distribution of Listeria monocytogenes (LM) in situ during the early phase of infection. Using an immunohistochemistry approach, five distinct phagocyte populations contained LM after i.v. challenge and accounted for roughly all bacterial signal in tissue sections. Our analysis showed that LM was initially captured by a wide range of phagocytes in the marginal zone, where the growth of LM appeared to be controlled. The cellular distribution of LM within phagocyte populations changed rapidly during the first few hours, decreasing in marginal zone macrophages and transiently increasing in CD11c(+) DC. After 4-6 h LM was transported to the periarteriolar lymphoid sheath where the infective foci developed and LM grew exponentially. PMID- 19130473 TI - Internalization of Dectin-1 terminates induction of inflammatory responses. AB - Dectin-1 is a pattern-recognition receptor recognizing beta-(1,3)-glucans found on fungal cell walls. Dectin-1 plays an important role in immunity to fungi by mediating phagocytic clearance of fungal particles and inducing transcription of innate response genes. We show here that the two processes are linked and that Dectin-1 signalling for inflammation is attenuated by phagocytosis. Blocking Dectin-1 ligand-dependent internalization using either actin polymerization or dynamin inhibitors, large non-phagocytosable beta-glucan particles or poorly phagocytic cells leads in all cases to enhanced and sustained activation of downstream signalling pathways and culminates in production of high levels of pro inflammatory cytokines. These findings establish the importance of phagocytosis not only in the clearance of pathogens, but also in the modulation of pattern recognition receptor signalling and strongly suggest that internalization is the first step to attenuation of Dectin-1-mediated pro-inflammatory responses. PMID- 19130475 TI - IL-4 blocks M-CSF-dependent macrophage proliferation by inducing p21Waf1 in a STAT6-dependent way. AB - Macrophages are recruited from the blood stream to the inflammatory loci to carry out their functional activities. In an early phase of the cell cycle, macrophages become activated by Th1-type cytokines (i.e. IFN-gamma), thereby producing several factors (cytokines, NO, etc.) and developing pro-inflammatory activities. When bacteria and apoptotic bodies are removed, through the interaction with Th2 type cytokines (i.e. IL-4), macrophages become anti-inflammatory and repair damaged tissues. Incubation of bone-marrow-derived macrophages with IFN-gamma or IL-4 blocked their proliferation. While M-CSF withdrawal caused cell cycle arrest at the early G(1) phase, treatment of macrophages with IFN-gamma or IL-4 caused this arrest later, at the G(1)/S boundary. Proliferation arrest was not due to an induction of apoptosis. IFN-gamma and IL-4 induced the expression of the cyclin dependent kinase (Cdk) inhibitor p21(Waf1). Using KO mice and iRNA experiments, we found that p21(Waf1)is required for IL-4- but not for IFN-gamma-dependent inhibition of macrophage proliferation. IL-4 inhibited M-CSF-dependent Cdk-2 and Cdk-4 activities, which are necessary for entry and passage through the S phase of the cell cycle. The signal transduction used to induce the expression of p21(Waf1)after interaction of IL-4 with the corresponding receptor was mediated by STAT6. Thus, IL-4 and IFN-gamma blocked M-CSF-induced macrophage proliferation through distinct mechanisms. PMID- 19130476 TI - Neonatal exposure to staphylococcal superantigen improves induction of oral tolerance in a mouse model of airway allergy. AB - The hygiene hypothesis suggests that lack of microbial stimulation in early infancy may lead to allergy, but it has been difficult to identify particular protective microbial exposures. We have observed that infants colonised in the first week(s) of life with Staphylococcus aureus have lower risk of developing food allergy. As many S. aureus strains produce superantigens with T-cell stimulating properties, we here investigate whether neonatal mucosal exposure to superantigen could influence the capacity to develop oral tolerance and reduce sensitisation and allergy. BALB/c mice were exposed to staphylococcal enterotoxin A (SEA) as neonates and fed with OVA as adults, prior to sensitisation and i.n. OVA challenge. Our results show that SEA pre-treated mice are more efficiently tolerised by OVA feeding, as shown by lower lung-cell infiltration and antigen specific IgE response in the SEA pre-treated mice, compared with sham-treated mice. This was not due to deletion or anergy of lymphocytes by SEA treatment, because the SEA pre-treated mice that were fed with PBS showed similar inflammatory response as the sham-treated PBS-fed mice. Our results suggest that strong T-cell activation in infancy conditions the mucosal immune system and promotes development of oral tolerance. PMID- 19130477 TI - PPP1R9B (Neurabin 2): involvement and dynamics in the NK immunological synapse. AB - The NK immunological synapse (NKIS) is a dynamic structure dependent on the assembly of membrane, cytoskeletal and signaling components. These serve to focus and generate stimuli for adhesion and orientation of the cytoskeleton for targeted cytolytic granule release. Previous studies have demonstrated the importance of the cytoskeleton in these processes. We previously identified PPP1R9B (neurabin 2, spinophilin) as a cytoskeletal component of the NK-like cell line YTS. We demonstrate that (i) PPP1R9B gradually accumulates at the NKIS in a maturation stage-dependent manner; (ii) it mimics the early kinetics of actin recruitment to the NKIS but it precedes actin departure from the site; (iii) it is recruited by CD18 stimulation but not by CD28 ligation; (iv) it is required for the maintenance of the cortical F-actin organization in the YTS cells and knocking down PPP1R9B reduces the frequency of YTS-target cell conjugation, possibly due to the collapsed F-actin cytoskeleton in these cells. These results indicate that PPP1R9B is required for synapse formation in the NK cells and suggest that it may be involved in the maintenance of cellular architecture by regulation of actin assembly, possibly acting to stabilize the NKIS until granule release is eminent. PMID- 19130478 TI - Long-lasting cross-presentation of tumor antigen in human DC. AB - DC cross-present exogenous antigens on MHC class I molecules, a process required for the onset of anti-tumor immune responses. In order to study the cross presentation of tumor antigens by human DC, we compared the pathways of cross presentation of long peptides requiring internalization and intracellular processing with the direct presentation of short peptides, which does not require intracellular processing. We found that, after brief incubations with DC, short peptides were presented to CD8(+) T cells with higher efficiencies than long peptides. After longer times of chase in the absence of peptide, however, the efficiency of presentation of the two types of peptides was reversed. After 2-3 days, DC pulsed with long peptides still activated T cells efficiently, while DC pulsed with short peptides failed to do so. Long-lasting presentation of the long peptides was, at least in part, due to a stored persistent pool of antigen, which was still available for loading on MHC class I molecules after several days of chase. These results show that the use of long synthetic peptides allows the efficient, long-lasting, presentation of tumor antigens, suggesting that long peptides represent an interesting approach for active anti-tumor vaccination. PMID- 19130479 TI - Captive cotton-top tamarins' (Saguinus oedipus oedipus) use of landmarks to localize hidden food items. AB - Seventeen captive cotton-top tamarins (Saguinus oedipus oedipus) were individually tested on their use of spatial relationships between landmarks to locate multiple hidden food items. In two experiments, the tamarins were presented with a spatial-foraging task in which positions of hidden food rewards were fixed in relation to an array of visual cues. In Experiment 1, the cues+hidden food configuration was rotated 90 degrees and the tamarins were successful in locating the food items significantly above chance levels (P<0.01). In Experiment 2 the cues+hidden food configuration was translated (up, down or sideways) from the previously learned configuration, and the monkeys successfully localized the hidden food items (P<0.001). Results indicate that the tamarins relied on the spatial relationship between the multiple landmarks to locate hidden food items rather than on an associative or beacon strategy. The results of these experiments support the contention that when contextually appropriate these captive New World monkeys have the capacity to rely on the spatial relationship or positions of several cues as an array to localize points in their environment. PMID- 19130480 TI - In vivo analysis of MT-based vesicle transport by confocal reflection microscopy. AB - The use of confocal reflection microscopy (CRM) for the in vivo analysis of microtubule (MT) mediated transport of lipid droplets in the developing Drosophila egg primordia is described here. The developing Drosophila oocytes are ideal objects to study MT-mediated transport in vivo: transport of e.g. the lipid droplets can be conveniently, selectively and sensitively monitored through CRM and the egg primordia are readily available for physical, chemical and/or genetic manipulations. CRM is a non-destructive way to follow vesicle movement and allows high frame rate image recording. When combined with fluorescence imaging, CRM offers simultaneous visualization of the cargo and the protein(s) of interest, i.e. a motor or a cargo adapter, thus allowing a better understanding of MT mediated transport and spatiotemporal coordination of the transport machinery. PMID- 19130481 TI - 3D image stack reconstruction in live cell microscopy of Drosophila muscles and its validation. AB - Rapid movements of live tissues during the acquisition of 3D image stacks can result in misalignments between successive image slices. The remodeling of the muscles in Drosophila metamorphosis is an example where sporadic motion during image acquisition impede image analysis and volume visualization. Most of the image stack registration algorithms applied in microscopy are aimed at the linear alignment of fixed histological sections. However, live muscles are nonrigid objects and their contractions and relaxations represent nonlinear transformations that cannot be properly rectified by applying purely linear registration methods. We developed a fully automated area-based nonrigid stack registration (NSR) method that minimizes the mean square error of intensities between successive image slices. The mapping function is formulated using the thin plate spline (TPS). A hierarchical linear to nonlinear, coarse to fine matching strategy is applied to ensure stability and fast convergence. Topological structure is preserved by constraining the step size of the nonlinear transformation. To assess the accuracy of 3D reconstruction, we propose a new benchmarking method that measures geometrical features of restored nuclei. We tested our algorithm on image stacks generated by laser scanning confocal microscopy that show live muscles during the prepupal stage of Drosophila metamorphosis. Our registration algorithm is able to restore image stacks that are distorted by periodic contraction of muscles. Quantitative assessment of registration performance agrees well with qualitative visual inspection. Our NSR method is able to restore image stacks for the purpose of visualization and quantitative analysis of Drosophila metamorphosis and, potentially, various other processes in developmental biology studied by 3D live cell microscopy. PMID- 19130482 TI - Absolute CD4+ cell count using a plastic microchip and a microscopic cell counter. AB - We have designed and evaluated the performance of a simple, rapid, and affordable method for counting CD4(+) T-cells with the use of plastic microchips. This new system is an adaptation of a "no-lyse, no-wash," volumetric single platform assay, and absolute CD4(+) counts are determined with the use of a microscopic scanning cell counter. To assess the CD4(+) count test precision and linearity of the system, measured CD4(+) counts were compared with two other reference assays (single and dual platform flow cytometry) with the use of 123 clinical samples including samples obtained from 35 HIV-infected patients, and artificially diluted samples. A correlation between the results from the use of the new method and from the use of the two other reference assays was r = 0.98 for the clinical samples. A dilution test of the new method demonstrated a linearity of r >or= 0.99, with coefficients of variation FE(RH) phase transition occurred. PMID- 19130613 TI - Comparative ultrasound evaluation of human trabecular bone graft properties after treatment with different sterilization procedures. AB - New sterilization methods for human bone are likely to affect the mechanical properties of human cancellous grafts. These mechanical properties dictate the short- and mid-term results of the orthopedic procedure. The aim of this study was to compare the effects on bone mechanical properties, as assessed by ultrasound velocity, of different sterilization methods used under similar conditions: bleach and sublimation, humid heat, successive baths of physiological saline with osmotic detersion, and CO(2) in the supercritical phase. Alterations in mechanical properties were small with CO(2) (velocity change: -2%) and humid heat (-2.5%). Osmotic detersion had a significant but moderate effect (-4.7%). The -9% change with the protocol involving bleach suggested a greater than 30% decrease in load to failure, based on earlier studies. Gamma irradiation of defatted trabecular allografts, in a dose of 10 or 25 KGy, produced no significant changes in ultrasound velocity. Powerful protein denaturants used in sterilization protocols substantially alter the mechanical resistance of the grafts, which may jeopardize the orthopedic procedure. PMID- 19130614 TI - A lubricious formulation exhibiting reduced thrombogenicity, cell proliferation, and protein adsorption. AB - The adhesion of human platelets, erythrocytes, and leukocytes, the adsorption of protein, and the proliferation of human umbilical vein endothelial cells (HUVEC) on the surface of electropolished stainless steel and the lumen of polyurethane tubing coated with Hydromer's lubricious Duality T8B formulation was evaluated. Following exposure to a platelet-enriched suspension from citrated human whole blood, stainless steel coated with this formulation exhibited significantly reduced adhesion of platelets, erythrocytes, and granulocytes. This reduction in adhesion was confirmed using an immunohistochemical method utilizing antibodies to CD41, CD235, and CD15, respectively. The proliferation of HUVEC cells were significantly reduced when cultured on coated stainless steel. This formulation was also able to significantly reduce the adsorption of plasma proteins and the major protein in tear fluid (lysozyme) to the surface of stainless steel. The nonthrombogenic properties of Duality T8B after application to the lumen of polyurethane tubing were also examined. Following a short-term (3 h) static exposure to citrated human whole blood, microscopic examination revealed that the adhesion of platelets and erythrocytes was reduced significantly, a finding confirmed using anti-CD41 and anti-CD235 antibodies in the immunohistochemical method. A long-term (12 day) study yielded essentially identical results indicating a significant reduction in the adhesion of blood components on the luminal surface of coated polyurethane tubing. In summary, these data indicate that the application of Duality T8B onto surfaces of medical devices, such as catheters, extracorporeal circuitry, and coronary stents, could aid in reducing or preventing not only thrombus formation but also the process of restenosis. PMID- 19130615 TI - Orienting neurite growth in electrospun fibrous neural conduits. AB - In cases where direct suture of two nerve stumps is not feasible, generally a scaffold is required to bridge the gap and guide nerve regeneration. The scaffold used in nerve tissue engineering applications requires optimal guidance effect, mechanical strength, and cellular compatibility. A number of studies suggest that fibrous scaffolds can potentially be used for nerve regeneration, as the aligned fibers can provide the guidance effect for axonal growth and the fibrous structure mimics the nerve microenvironment. A practical method to fabricate the fibrous nerve conduit with structural guidance cue for neurite growth will improve its potential application in neural tissue engineering. In this study, fibrous tubular scaffolds were fabricated using an electrospinning technique. The fibrous conduit was composed of aligned fibers in the interior layer and randomly oriented fibers in the exterior layer. In order to investigate the fiber diameter on neurite extension and directional growth, fibers with different diameter in the subcellular size range were fabricated and the neurite growth on these fibers was investigated. We observed that PC12 cells' neurites showed similar parallel growth on the aligned fibers irrespective of fiber diameter. Neurite length on aligned fibers, with fiber diameters of 3.7 +/- 0.5 microm and 5.9 +/- 0.9 microm, was significantly longer than neurite length on randomly oriented fibers. This study showed that electrospinning provides a practical solution to fabricate fibrous nerve conduit by controlling the conduit inner diameter and fiber alignment and, hence, potentially improves the application of the conduit in neural regeneration. PMID- 19130616 TI - Inflammatory bowel disease patients who leave hospital against medical advice: predictors and temporal trends. AB - BACKGROUND: Leaving hospital against medical advice (AMA) may have consequences with respect to health-related outcomes; however, inflammatory bowel disease (IBD) patients have been inadequately studied. Thus, we determined the prevalence of self-discharge, assessed predictors of AMA status, and evaluated time trends. METHODS: We analyzed the 1995-2005 Nationwide Inpatient Sample (NIS) to identify 93,678 discharges with a primary diagnosis of IBD admitted to the hospital emergently and did not undergo surgery. We described the proportion of IBD patients who left AMA. Predictors of AMA status were evaluated using a multivariate logistic regression model and temporal trend analyses were performed with Poisson regression models. RESULTS: Between 1995 and 2005, 1.31% of IBD patients left hospitals AMA. Crohn's disease (CD) patients were more likely to leave AMA (adjusted odds ratio [aOR], 1.53; 95% confidence intervals [CI]: 1.30 1.79). Characteristics associated with leaving AMA included: ages 18-34 (aOR, 7.77, 95% CI: 4.34-13.89); male (aOR, 1.75; 95% CI: 1.55-1.99); Medicaid (aOR, 4.55; 95% CI: 3.81-5.43) compared to private insurance; African Americans (aOR, 1.34; 95% CI: 1.09-1.64) compared to white; substance abuse (aOR, 2.75; 95% CI: 2.14-3.54); and psychosis (aOR, 1.55; 95% CI: 1.13-2.14). The incidence rates of self-discharge for CD patients were stable (P > 0.05) between 1995 and 1999, while they significantly (P < 0.0001) increased after 1999. In contrast, AMA rates for UC patients remained stable during the study period. CONCLUSIONS: Approximately 1 in 76 IBD patients admitted emergently for medical management leave the hospital AMA. These were primarily disenfranchised patients who may lack adequate outpatient follow-up. PMID- 19130617 TI - Extracorporeal photopheresis for the treatment of refractory Crohn's disease: results of an open-label pilot study. AB - BACKGROUND: Extracorporeal photopheresis (ECP) is effective in immune-mediated disorders. A prospective, uncontrolled pilot study was conducted to evaluate the safety and efficacy of ECP in patients with active Crohn's disease (CD) who were refractory to or intolerant of immunosuppressants and/or anti-TNF therapies. METHODS: Patients with moderate-to-severely active CD (Crohn's Disease Activity Index [CDAI] 220-450 points) underwent 12 weeks of ECP treatment (Weeks 1-4: twice weekly, every week; Weeks 5-12: twice weekly, every other week). Clinical response was defined as a decrease in the CDAI of >or=100 points or remission (CDAI <150 points) at Week 12. Patients who responded at Week 12 could receive an additional 12 weeks of ECP treatment (twice weekly, every other week) in an extension study. RESULTS: Twenty-eight patients were enrolled with a mean baseline CDAI score of 314 (range 207-457). At Week 12, 14 patients (50%) responded; 13 patients responded within 6 weeks. Seven patients (25%) attained remission by Week 12. Three of 5 patients with open fistulae at baseline had fistula closure. Response was similar among patients naive to anti-TNF agents and patients who had previously been refractory or intolerant to anti-TNF agents. Of the 12 patients who entered the extension study, 9 (75%) maintained their response at Week 24. CONCLUSIONS: In patients with moderate-to-severely active CD who were refractory to or intolerant of immunosuppressants and/or anti-TNF agents, ECP was well tolerated and induced clinical response (50%) and remission (25%) in patients. Most patients were able to maintain a response with continued treatments. PMID- 19130618 TI - Role of PET and combination PET/CT in the evaluation of patients with inflammatory bowel disease. AB - BACKGROUND: Positron emission tomography (PET) is a nuclear imaging technique providing noninvasive, three-dimensional, whole-body, quantitative images. The primary use of PET is in tumor detection and staging. More recently, it has been shown to be of value in assessing patients with inflammatory processes. To date the role of PET in the management of patients with inflammatory bowel disease (IBD) has not been defined. METHODS: The electronic literature (August 1966 to June 2008) on the utilization of PET in IBD was reviewed. Further references were obtained by cross-referencing from key articles. RESULTS: There have been no randomized studies to date examining the role of PET in the management of patients with IBD. Comparative studies have demonstrated a high degree of correlation between PET-detected segmental (18)F-fluorodeoxyglucose uptake and sites of macroscopic and histological inflammation noted on endoscopy. Prospectively, PET performs favorably in comparison to conventional imaging modalities for IBD such as immunoscintigraphy, magnetic resonance imaging (MRI), and hydro-MRI. Several case series have highlighted the utility of PET in the diagnosis of IBD in the pediatric population. The recent development of PET/computed tomography (CT) combines the physiological sensitivity of PET with the anatomical accuracy of CT, increasing the specificity of PET. CONCLUSIONS: The case series and nonrandomized studies published to date emphasize the utility of PET in assessing patients with IBD and justify further study. The development of PET/CT represents a significant advance and should be considered in the evaluation of patients with IBD. PMID- 19130620 TI - Words fail me: the verbal IQ deficit in inflammatory bowel disease and irritable bowel syndrome. AB - BACKGROUND: Many chronic illnesses are accompanied by impaired cognitive functioning. In people with Inflammatory Bowel Disease (IBD), there is some research to suggest a decrement in verbal IQ (VIQ), when compared to people with Irritable Bowel Syndrome (IBS) and healthy controls. Although this is an important finding, it is necessary to ensure that such deficits are not due to methodological problems such as the failure to take into account pre-morbid functioning. METHODS: A total of 88 people (IBD, N = 29; IBS, N = 29; Controls, N = 30) completed the Wechsler Abbreviated Scale of Intelligence (WASI), the Wechsler Test of Adult Reading (WATR), the Trait Rumination Questionnaire (TRQ), the Center for Epidemiologic Studies Depression Scale (CES-D), and the General Health Questionnaire (GHQ-12). RESULTS: We found evidence of a VIQ decrement in both IBD and IBS groups when measured against both healthy controls and against their own pre-morbid IQ scores (WTAR-Predicted WAIS-III IQ measures). However, the decrement was larger (and of clinical significance) in the IBD group but not in the IBS group. CONCLUSION: Some tentative evidence is presented which suggests that poor VIQ performance may be due in part to interference from excessive rumination. PMID- 19130619 TI - Pain and inflammatory bowel disease. AB - Abdominal pain is a common symptom of inflammatory bowel disease (IBD: Crohn's disease, ulcerative colitis). Pain may arise from different mechanisms, which can include partial blockage and gut distention as well as severe intestinal inflammation. A majority of patients suffering from acute flares of IBD will experience pain, which will typically improve as disease activity decreases. However, a significant percentage of IBD patients continue experiencing symptoms of pain despite resolving inflammation and achieving what appears to be clinical remission. Current evidence suggests that sensory pathways sensitize during inflammation, leading to persistent changes in afferent neurons and central nervous system pain processing. Such persistent pain is not only a simple result of sensory input. Pain processing and even the activation of sensory pathways is modulated by arousal, emotion, and cognitive factors. Considering the high prevalence of iatrogenic as well as essential neuropsychiatric comorbidities including anxiety and depression in IBD patients, these central modulating factors may significantly contribute to the clinical manifestation of chronic pain. The improved understanding of peripheral and central pain mechanisms is leading to new treatment strategies that view pain as a biopsychosocial problem. Thus, improving the underlying inflammation, decreasing the excitability of sensitized afferent pathways, and altering emotional and/or cognitive functions may be required to more effectively address the difficult and disabling disease manifestations. PMID- 19130622 TI - Gauging cell phones' possible risk. PMID- 19130623 TI - Headache. In this issue. PMID- 19130621 TI - Effects of acute agonist treatment on subcellular distribution of kappa opioid receptor in rat spinal cord. AB - We investigated whether acute treatment with agonists affected the subcellular distribution of kappa opioid receptor (KOPR) in the dorsal horn of the rat lumbar spinal cord by using immunoelectron microscopy. Rats were injected intrathecally (i.t.) with U50,488H (100 nmole), dynorphin A(1-17) (15 nmole), or vehicle. The doses chosen have been shown to induce antinociception. Rats were perfused transcardially 30 min later, and lumbar spinal cords were removed and processed for electron microscopic analysis. KOPR was stained with KT-2, a specific polyclonal antibody against the rat/mouse KOPR(371-380) peptide, followed by gold labeled secondary antibody and silver intensification. The silver grains were present in axons, terminals, dendrites, and somata, and the association with plasma membranes was quantified in dendrites, because KOPR immunoreactivity was most frequently observed in these profiles. In vehicle-treated rats, approximately 27% of KOPR immunoreactivity was associated with plasma membranes. U50,488H, i.t., did not cause a significant change in the percentage of KOPR present on plasma membranes, whereas dynorphin A, i.t., significantly decreased cell surface KOPR to approximately 19%. In summary, these data indicate that U50,488H and dynorphin A differentially regulate the subcellular distribution of endogenous KOPR. PMID- 19130625 TI - [Debate and cancer]. PMID- 19130624 TI - Identification of quantitative trait loci for locomotor activation and anxiety using closely related inbred strains. AB - We carried out a quantitative trait loci (QTL) mapping experiment in two phenotypically similar inbred mouse strains, C57BL/6J and C58/J, using the open field assay, a well-established model of anxiety-related behavior in rodents. This intercross was initially carried out as a control cross for an ethylnitrosurea mutagenesis mapping study. Surprisingly, although open-field behavior is similar in the two strains, we identified significant QTL in their F2 progeny. Marker regression identified a locus on Chr 8 having associations with multiple open-field measures and a significant interaction between loci on Chr 13 and 17. Together, the Chr 8 locus and the interaction effect form the core set of QTL controlling these behaviors with additional loci on Chr 1 and 6 present in a subset of the behaviors. PMID- 19130626 TI - [A 51-year-old woman with acute chest pain]. PMID- 19130627 TI - [When chest pain is not myocardial infarction]. PMID- 19130628 TI - Development-dependent expression of calreticulin in the brain and other tissues of the Japanese monkey, Macaca fuscata. AB - BACKGROUND: Calreticulin (CRT) is a molecular chaperone localized to endoplasmic reticulum lumen. The protein is involved in the correct folding of glycoproteins and its cellular level changes depending on various physiologic conditions. METHODS: To clarify the Crt level in various tissues of the Japanese monkey during postnatal development, an enzyme-linked immunosorbent assay system that we have established was applied. RESULTS AND CONCLUSIONS: Calreticulin is distributed ubiquitously in various tissues. Its level in the heart, lung, liver, spleen, and kidney was high in newborns and decreased in juveniles and adults. In various cerebral areas, Crt was present in the gray matter but scarcely found in the white matter. The Crt levels in the cerebral areas were low in newborns and increased in juveniles and adults. These distribution and developmental changes in Crt might be correlated with the quality control of glycoproteins that are synthesized in respective tissues. PMID- 19130629 TI - [Health insurance means better health or Rwanda's population]. PMID- 19130630 TI - Eva Peron: cervical cancer and the effect on a nation. PMID- 19130631 TI - Role of MRI of the breast in the evaluation of the symptomatic patient. AB - PURPOSE OF REVIEW: The role of MRI in breast pathology is still controversial. The technique has a high sensitivity for detection of breast carcinoma, but specificity is low. Therefore, it is only useful for right indications. RECENT FINDINGS: Studies are still performed to define the role of MRI. For diagnosis of a breast lesion, image-guided percutaneous core biopsy is the method of choice, and MRI is not indicated. It has a place in the detection of carcinoma in patients with nipple discharge. MRI has a role in the search of an occult primary breast carcinoma in patients with metastasis and primary unknown cancer. MRI is the best technique for locoregional staging of breast carcinoma. Its role in detection of recurrence is controversial, as image-guided biopsy has to be done if a suspicious lesion is seen. In dense breasts or difficult scar formation, MRI is able to detect recurrence. MRI is sensitive in detection of implant rupture in symptomatic augmented patients. SUMMARY: MRI has a role in well defined indications, but its role is still controversial even in some of these indications. MRI is not indicated in diagnosis in which image-guided percutaneous biopsy is the method of choice. Further studies must be performed. PMID- 19130633 TI - Optimizing follow-up after breast cancer treatment. AB - PURPOSE OF REVIEW: This review focuses on studies that help elucidate the optimum approach to posttreatment follow-up of breast cancer patients. RECENT FINDINGS: The re-conceptualization of follow-up under the rubric of survivorship care and the benefit of survivorship care plans, studies on the elements of follow-up care including surveillance mammograms, alternative models of follow-up care including primary care based follow-up, and patterns of care studies that involve population-based samples are discussed. Posttreatment follow-up of breast cancer patients continues to be controversial despite almost two decades of research. The research does show that surveillance mammograms are beneficial and guidelines recommend routine surveillance mammograms annually. Other routine surveillance tests are not beneficial and are not recommended. The precise frequency and duration of clinical visits is not known and recommendations vary; but most do support continued clinical assessment. Alternative models, such as nurse-led follow-up or less frequent follow-up, show good patient satisfaction and quality of-life outcomes. Primary care based followup results in similar clinical and quality-of-life outcomes as specialist-based follow-up. SUMMARY: Instead of trying to find a one-size-fits-all approach, the focus should be on an individualized tailored approach in which the patient makes an informed decision on the basis of evidence of actual benefits and risks. PMID- 19130634 TI - Calcium homeostasis in plant cell nuclei. AB - In plant cells, calcium-based signaling pathways are involved in a large array of biological processes, including cell division, polarity, growth, development and adaptation to changing biotic and abiotic environmental conditions. Free calcium changes are known to proceed in a nonstereotypical manner and produce a specific signature, which mirrors the nature, strength and frequency of a stimulus. The temporal aspects of calcium signatures are well documented, but their vectorial aspects also have a profound influence on biological output. Here, we will focus on the regulation of calcium homeostasis in the nucleus. We will discuss data and present hypotheses suggesting that, while interacting with other organelles, the nucleus has the potential to generate and regulate calcium signals on its own. PMID- 19130632 TI - Diet and breast cancer prognosis: making sense of the Women's Healthy Eating and Living and Women's Intervention Nutrition Study trials. AB - PURPOSE OF REVIEW: To clarify the role of dietary pattern on prognosis in breast cancer survivors. RECENT FINDINGS: Observational trials show mixed results that do not strongly support an independent role for dietary pattern in prognosis. Women's Intervention Nutrition Study and Women's Healthy Eating and Living (WHEL) are two large randomized controlled trials that address this question. The interventions from both studies achieved significant reductions in energy from fat, and the WHEL Study achieved large increases in vegetables, fruit and fiber. Women's Intervention Nutrition Study examined postmenopausal women only and reported a not-quite-significant improved prognosis for women in the intervention group, with the benefit focused on ipsilateral localized recurrences, but little improvement in the more important distal recurrences. This review considers only WHEL postmenopausal women to aid a direct comparison with Women's Intervention Nutrition Study. The WHEL Study reported a convincing lack of association between diet and prognosis. However, a secondary analysis suggests that the dietary intervention reduced distal recurrences among the subgroup without hot flashes at baseline. SUMMARY: There is no convincing evidence that changing dietary pattern following breast cancer diagnosis will improve prognosis for most women with early stage breast cancer. However, it would appear to be important for some subgroups. Further investigation of mechanisms for such selective action is needed. PMID- 19130635 TI - Does auricular acupuncture reduce postoperative vomiting after cholecystectomy? AB - PURPOSE: To find the efficacy of auricular acupuncture for the prevention of postoperative nausea and vomiting after cholecystectomy. MATERIALS AND METHODS: One hundred (100) female patients undergoing transabdominal cholecystectomy were randomly allocated to two groups of 50 (auricular acupuncture treatment group and nontreatment group) in order to test the effectiveness of auricular acupuncture. Statistical significance (p < 0.05) was determined using an unpaired t-test for age, weight, height, and duration of anesthesia; a chi-square test was used to analyze the incidence of vomiting. RESULTS: There was no significant difference in age, weight, height, or duration of anesthesia among the two groups of patients. There was a significant difference between the control and auricular acupuncture treatment groups in the incidence of vomiting 24 hours after surgery (66% and 0%, respectively, p < 0.01). No noteworthy side effects from treatment were observed. CONCLUSION: Auricular acupuncture is effective in reducing vomiting following transabdominal cholecystectomy in female patients. PMID- 19130636 TI - Comparison between different off-season resistance training programs in Division III American college football players. AB - The purpose of this study was to examine the efficacy of periodization and to compare different periodization models in resistance trained American football players. Fifty-one experienced resistance trained American football players of an NCAA Division III football team (after 10 weeks of active rest) were randomly assigned to 1 of 3 groups that differed only in the manipulation of the intensity and volume of training during a 15-week offseason resistance training program. Group 1 participated in a nonperiodized (NP) training program, group 2 participated in a traditional periodized linear (PL) training program, and group 3 participated in a planned nonlinear periodized (PNL) training program. Strength and power testing occurred before training (PRE), after 7 weeks of training (MID), and at the end of the training program (POST). Significant increases in maximal (1-repetition maximum [1RM]) squat, 1RM bench press, and vertical jump were observed from PRE to MID for all groups; these increases were still significantly greater at POST; however, no MID to POST changes were seen. Significant PRE to POST improvements in the medicine ball throw (MBT) were seen for PL group only. The results do not provide a clear indication as to the most effective training program for strength and power enhancements in already trained football players. Interestingly, recovery of training-related performances was achieved after only 7 weeks of training, yet further gains were not observed. These data indicate that longer periods of training may be needed after a long term active recovery period and that active recovery may need to be dramatically shortened to better optimize strength and power in previously trained football players. PMID- 19130637 TI - Long-term power performance of elite Australian rules football players. AB - Measuring and tracking performance variables such as peak power output is essential for assessing the effects of training and for informing adjustments to optimize program design. The purpose of this study was to track the long-term changes in muscular power, velocity, and jump height of elite Australian Rules Football (ARF) players during a 3-year period. Twelve members of an ARF team were tracked during the study. The physical characteristics of the subjects were age, 25.3 6 2.8 years; body mass, 93.0 6 6.8 kg; and height, 192 6 6 cm. The subjects performed unweighted and weighted (40 kg) countermovement jumps (CMJ) and static jumps (SJ). Peak power, jump height, and bar velocity were determined using a force plate and position transducer.Measures of peak power output during both the CMJ and SJ indicated significant (p# 0.001) increases of 13% (effect size [ES] = 0.70) and 17% (ES = 1.77), respectively, during the course of the tracking period. There was also a significant 9% (ES = 0.71) increase in CMJ40 peak power. The subjects' CMJ peak velocity improved by 10.7% (ES = 0.34), and SJ peak velocity improved by 12.6% (ES = 0.37). The data from this study show that elite level ARF players can continue to increase muscular power and velocity. This information is of interest to strength and conditioning coaches who are interested in improving power performance of their athletes for a long-term period. PMID- 19130638 TI - Lateralisation of spatial processing and age. AB - Studies assessing spatial ability suggest right hemisphere specialisation for coordinate spatial processing and left hemisphere specialisation for categorical spatial processing. With regard to healthy ageing, spatial abilities may be affected selectively, with right hemisphere based coordinate processes being more vulnerable to age-related decline, but previous research has been inconsistent. In the present study, age and hemispheric specialisation of categorical and coordinate spatial abilities were explored. Testing 56 right-handed younger and older participants clearly showed a left hemisphere advantage for the categorical task and a right hemisphere advantage for the coordinate spatial task, for both age groups combined. Older adults were slower to process information and make a spatial judgement; nevertheless, the neural specialisation underlying spatial abilities seems to have remained consistent with age. PMID- 19130639 TI - Eight weeks of resistance training can significantly alter body composition in children who are overweight or obese. AB - The purpose of this study was to investigate the effect of an 8-week resistance training program on children who were overweight or obese. Forty-eight children (n = 26 girls and 22 boys; mean age = 9.7 years) participated in an 8-week undulating periodized resistance training program for 3 d x wk-1. Measures of body composition via dual-energy X-ray absorptiometry, anthropometry, strength, and power were made before and after the training intervention. There was a significant decrease in absolute percent body fat of 2.6%(p = 0.003) and a significant increase in lean body mass of 5.3% (p = 0.07). There were no significant changes in height, weight, body mass index, total fat mass, or bone mineral content. There were significant increases in 1-repetition maximum squat (74%), number of push-ups (85%), countermovement jump height (8%), static jump height (4%), and power (16%). These results demonstrate that the resistance training program implemented produces significant changes in body composition and strength and power measures, as well as being well tolerated by the participants. An undulating periodized program provides variation and significantly increases lean body mass, decreases percent body fat, and increases strength and power in children who are overweight and obese. PMID- 19130640 TI - Handedness, eyedness, and hand--eye crossed dominance in patients with schizophrenia: sex-related lateralisation abnormalities. AB - Schizophrenia is referred to as cerebral lateralisation abnormality. In this study the possible relationships among handedness, eye dominance, and crossed and non congruent hand-eye dominance in patients with schizophrenia are investigated. A total of 88 patients with schizophrenia and 118 controls were included in the study.The patient group included 60 men and 28 women who ranged in age from 17 to 63 years. Diagnoses were made on the basis of information provided from clinical interviews and Structured Clinical Interview for DSM-IV. Handedness was ascertained by using the Edinburgh Handedness Inventory. Eye dominance was measured only by the near-far alignment test. Patients with schizophrenia had a significantly increased frequency of mixed-handedness and decreased frequency of both right- and left-handedness in comparison with controls. Also, the male patients with schizophrenia had significantly increased frequencies of left eye dominance, crossed hand-eye dominance, and non-congruent hand-eye dominance compared to controls, but not the female patients. Cerebral lateralisation abnormalities in schizophrenia may be associated with sex-related hormonal factors. PMID- 19130641 TI - Comparison of methods to quantify volume during resistance exercise. AB - The purpose of this investigation was to compare 4 different methods of calculating volume when comparing resistance exercise protocols of varying intensities. Ten Appalachian State University students experienced in resistance exercise completed 3 different resistance exercise protocols on different days using a randomized, crossover design, with 1 week of rest between each protocol. The protocols included 1) hypertrophy: 4 sets of 10 repetitions in the squat at 75% of a 1-repetition maximum (1RM) (90-second rest periods); 2) strength: 11 sets of 3 repetitions at 90% 1RM (5-minute rest periods); and 3) power: 8 sets of 6 repetitions of jump squats at 0% 1RM (3-minute rest periods). The volume of resistance exercise completed during each protocol was determined with 4 different methods: 1) volume load (VL) (repetitions [no.] x external load [kg]); 2) maximum dynamic strength volume load (MDSVL) (repetitions [no.] x [body mass- shank mass (kg) + external load (kg)]); 3) time under tension (TUT) (eccentric time +milliseconds] + concentric time +milliseconds]); and 4) total work (TW) (force [N] x displacement [m]). The volumes differed significantly (p , 0.05) between hypertrophy and strength in comparison with the power protocol when VL and MDSVL were used to determine the volume of resistance exercise completed. Furthermore, significant differences in TUT existed between all 3 resistance exercise protocols. The TW calculated was not significantly different between the 3 protocols. These data imply that each method examined results in substantially different values when comparing various resistance exercise protocols involving different levels of intensity. PMID- 19130642 TI - Acute physiological responses and time-motion characteristics of two small-sided training regimes in youth soccer players. AB - The purpose of this study was to examine the acute physiological responses and time-motion characteristics associated with continuous and intermittent small sided games (SSGs). The continuous (SSGC) regime involved 24 minutes' playing duration (no planned rest intervals), whereas the intermittent regime (SSGI) involved 4 x 6-minute bouts with 1.5 minutes of passive planned rest (work:rest ratio 4:1). Both training regimes were implemented across 3 SSG formats, which included games with 2 vs. 2, 4 vs. 4, and 6 vs. 6 players. Sixteen men's soccer players (mean +/- SE: age = 16.2 +/- 0.2 years, height = 173.7 +/- 2.1 cm, body mass = 65.0+/- 2.5 kg, estimated VO2max = 54.8 +/- 0.7 ml x kg-1 x min-1) participated in the study. Heart rate (HR) was measured every 5 seconds during all SSGs. Global ratings of perceived exertion (RPEs) were recorded immediately after the SSGs using the Borg scale (RPEs, 6-20). Capillary blood samples were drawn at rest and within 5 minutes after the end of each SSG. Time-motion characteristics were measured using portable global positioning system units. There were no significant differences between SSGC and SSGI for total distance covered or for distance traveled while walking, jogging, or running at moderate speed. However, players covered a significantly greater distance at 13.0-17.9 km x h-1, a greater total distance at higher running speed, and a greater total number of sprints (>18 km x h-1) with SSGI compared with SSGC. In contrast, global RPE and %HRmax were significantly higher in SSGC than in SSGI. Both intermittent and continuous SSG training regimes could be used during the season for match-specific aerobic conditioning. However, both training regimes used in this study seem unlikely to provide a sufficient stimulus overload for fully developing VO2max. PMID- 19130643 TI - Effect of creatine supplementation during cast-induced immobilization on the preservation of muscle mass, strength, and endurance. AB - Muscle and strength loss will occur during periods of physical inactivity and immobilization. Creatine supplementation may have a favorable effect on muscle mass and strength independently of exercise. The purpose of this study was to determine the effects of creatine supplementation on upper limb muscle mass and muscle performance after immobilization. Before the study, creatine-naive men (n = 7; 18-25 years) were assessed for lean tissue mass (dual-energy X-ray absorptiometry), strength (1-repetition maximum [1RM] isometric single arm elbow flexion/extension), and muscle endurance (maximum number of single-arm isokinetic elbow flexion/extension repetitions at 60% 1RM). After baseline measures, subjects had their dominant or nondominant (random assignment) upper limb immobilized (long arm plaster cast) at 90 degrees elbow flexion. Using a single blind crossover design, subjects received placebo (maltodextrin; 4 x 5 gxd-1) during days 1-7 and creatine (4 x 5 gxd-1) during days 15-21. The cast was removed during days 8-14 and 22-29. The dependent measures of lean tissue mass, strength, and endurance were assessed at baseline, postcast, and after the study. During immobilization, compared with isocaloric placebo, creatine supplementation better maintained lean tissue mass (Cr +0.9% vs. PLA -3.7%, p < 0.05), elbow flexor strength (Cr -4.1% vs. PLA -21.5%, p < 0.05), and endurance (Cr -9.6% vs. PLA -43%, p < 0.05), and elbow extensor strength (Cr -3.8% vs. PLA -18%, p < 0.05) and endurance (Cr -6.5% vs. PLA -35%, p < 0.05). These results indicate that short-term creatine supplementation attenuates the loss in muscle mass and strength during upper-arm immobilization in young men. PMID- 19130644 TI - Effects of instructional and motivational self-talk on the vertical jump. AB - The purpose of this study was to investigate the effects of instructional and motivational self-talk on performance and the kinematics of the vertical jump. After completing a 10-minute warm-up on a stationary bike, 12 men (mean +/- SD; 20.8+/- 3.0 years, 77.8 +/- 13.5 kg, 1.78 +/- 0.07 m) and 12 women (22.1 +/- 5.8 years, 62.6 +/- 6.7 kg, 1.65 +/- 0.05 m) performed 4 vertical jumps, 3 minutes apart, on a force plate set at a 1000-Hz sampling frequency. Before each trial, participants engaged in 1 of 4 counterbalanced interventions, verbalized out loud, which included motivational self-talk, instructional self-talk, neutral self-talk, or no instruction. One-way analysis of variance with repeated measures, followed by paired t-tests with a Bonferroni adjustment, were used to analyze data. Both instructional (0.415 m) and motivational (0.414 m) self-talk led to greater center-of-mass displacement than neutral self-talk (0.403 m, p = 0.001 and 0.003, respectively, alpha set at 0.008). Both instructional (263.9 N x s) and motivational self-talk (261.2 N x s) led to greater impulse than neutral self-talk (254.1 N x s, p = 0.005 and 0.004, respectively, alpha set at 0.025). Both instructional self-talk (582.6 degrees x s-1) and motivational self-talk (592.3 degrees x s-1) led to quicker angular rotation about the knee than neutral self-talk (565.8 degrees x s-1, p = 0.001 and 0.018, respectively, alpha set at 0.025). These results may indicate that self-talk leads to greater angular velocity about the knee, thus generating greater impulse and increased jump height-a conjecture that needs empirical testing. Self-talk may contribute to improved performance in sports requiring power-based skills. PMID- 19130645 TI - The influence of exercise order on blood lactate, perceptual, and affective responses. AB - The purpose of this study was to examine the influence of exercise order on blood lactate, perceptual, and affective responses to resistance exercise. Twenty-nine subjects (18 women, 11 men; 20.9 +/- 1.9 years) completed three sessions separated by a minimum of 48 hours. Session 1 determined the 10-repetition maximum (10RM) for nine resistance exercises. During sessions 2 and 3, exercises were completed in either a large to small or small to large muscle exercise order. The large to small muscle order was 1) chest press, 2) leg press, 3) rows, 4) leg extension, 5) overhead press, 6) hamstring curl, 7) biceps curl, 8) calve raise, and 9) triceps extension. Exercise order was reversed for the small to large condition. Participants performed two sets of each lift, with the first set being a warm-up at 80% 10RM, followed by one set at 100% 10RM with 1 minute of rest between each exercise. Rating of perceived exertion (RPE) was measured after completion of the second set. Blood lactate was recorded after exercises 1, 5, and 9. Affective measures were completed pre, during, post 0, and post 10 minutes. Lactate showed a significant time (p < 0.001) and condition x time interaction (p = 0.020). A significant difference was seen in average number of repetitions completed between sequences, with small to large performing more. There were no differences seen between exercise orders for average RPE. Analyses of Feeling Scale and Felt Arousal Scale scores showed only a significant main effect of time. A paired-sample t-test was conducted to examine differences in Feeling Scale for the two conditions at the different time points. Significant differences were found for Feeling Scale during exercise (after overhead press) and at post 10, with the small to large exercise order having greater Feeling Scale responses. No significant correlations were seen between blood lactate and perceptual or affective responses at any time point or in either exercise order. These findings may suggest that small to large exercise order may have beneficial physiological and psychological outcomes and potentially influence exercise adherence. PMID- 19130646 TI - The effect of stance width on the electromyographical activity of eight superficial thigh muscles during back squat with different bar loads. AB - Many strength trainers believe that varying the stance width during the back squat can target specific muscles of the thigh. The aim of the present work was to test this theory measuring the activation of 8 thigh muscles while performing back squats at 3 stance widths and with 3 different bar loads. Six experienced lifters performed 3 sets of 10 repetitions of squats, each one with a different stance width, using 3 resistances: no load, 30% of 1-repetition maximum (1RM), and 70% 1RM. Sets were separated by 6 minutes of rest. Electromyographic (EMG) surface electrodes were placed on the vastus medialis, vastus lateralis, rectus femoris, semitendinosus, biceps femoris, gluteus maximus, gluteus medium, and adductor maior. Analysis of variance and Scheffe post hoc tests indicated a significant difference in EMG activity only for the gluteus maximus; in particular, there was a higher electrical activity of this muscle when back squats were performed at the maximum stance widths at 0 and 70% 1RM. There were no significant differences concerning the EMG activity of the other analyzed muscles. These findings suggest that a large width is necessary for a greater activation of the gluteus maximus during back squats. PMID- 19130647 TI - Resistance exercise sessions do not provoke acute immunosuppression in older women. AB - It has been suggested that elderly people are more susceptible to infectious diseases because of immunosenescence. The well-described transient immunosuppressive effect of exercise may increase health risks in such populations. Although resistance training has been recommended to older individuals, little is known regarding its acute effects on immune indexes. This study examined the acute effects of resistance exercise sessions performed at different intensities (50 vs. 80% of one-repetition maximum [1RM]) on salivary immunoglobulin A (IgA), total leukocytes, lymphocytes and their subsets, and the cortisol levels of older women. Fifteen physically independent elderly women (mean age = 67.5 +/- 3.9 years) underwent, in a randomized manner, two experimental sessions comprising resistance exercise for the major muscular groups or one control session without exercise. Session 1 consisted of two sets of 13 repetitions at 50% 1RM, whereas session 2 consisted of two sets of eight repetitions at 80% 1RM. Blood and saliva samples were collected at rest, immediately after, and at 3 and 48 hours after the completion of the sessions. Blood cortisol, total leukocytes, lymphocytes and their subpopulations, and salivary IgA were measured by standard techniques. Overall, resistance exercise sessions failed to provoke significant decline on the examined immune function indexes. Instead, resistance exercise sessions induced significant elevation in salivary IgA levels, but this was not observed after the control session. Also, experimental sessions did not induce significant elevations in blood cortisol. It was concluded that resistance exercise sessions performed at 50 or 80% of 1RM do not present suppressive effects on the studied immune parameters. Conversely, a transient elevation was observed in salivary IgA levels, which requires support in future investigations. Taken together, the observed results suggest that elderly women can engage in resistance training programs without negative acute effects on the studied parameters of the immune system. PMID- 19130648 TI - Reliability and validity of a pressure algometer. AB - Algometers are devices that can be used to identify the pressure and/or force eliciting a pressure-pain threshold. It has been noted in pressure-pain threshold studies that the rate at which manual force is applied should be consistent to provide the greatest reliability. This study tested the reliability and construct validity of an algometer (1000-Hz sampling rate) by manually applying pressure on a force plate (500-Hz sampling rate): 10 sets of 5 applications to 80 N and 1 set of 5 applications to each force level: 20, 30, 40, 50, 60, 70, 80, 90, 100, and 110 N. The investigator had previously become familiar with and practiced with the algometer. The handheld algometer had a 1-cm2 round rubber application surface, and the maximum force reading was compared with maximum force readings by the force plate using SEM and t-tests. Force-time curves were analyzed for average slope representing rate of force application. Average Pearson (r) correlations between the maximum force reading of the algometer and force plate were excellent in both trials to 80 N (r = 0.990) and the incremental trials (r = 0.999). The application of force was reasonably constant, with slopes averaging 6.8 +/- 0.932 N x s-1. The SEE was 0.323 N. In conclusion, with previous familiarization and practice, an investigator may have high reliability in the rate of force application. The device itself was also highly correlated with readings from a force plate and, therefore, may be considered valid. PMID- 19130649 TI - How can research contribute to risk assessment? PMID- 19130650 TI - Soil protection in Europe. PMID- 19130651 TI - Implications of Clarias gariepinus (African catfish) propagation in Cuban waters. PMID- 19130652 TI - FL: RN won in gender/pregnancy discrimination: RN awarded trial in retaliatory termination case. PMID- 19130653 TI - Revitalizing environmental assessment. PMID- 19130654 TI - What is an appropriate level of protection? An example considering selenium exposures by aquatic birds. AB - Evaluating population-level risks to ecological receptors or developing toxicity thresholds intended to be protective of a population requires a population model to truly understand possible chemical-related impacts to the population of interest. For various reasons (e.g., lack of training in the application of population models to ecotoxicological questions), we often use laboratory-based (more common) or field-based (less common) toxicity data to develop a toxicity threshold that is assumed to be protective of population-level effects. Under this latter approach, an appropriate level of protection against exposure should focus on protecting the viability and productivity of populations of organisms, that is, maintaining approximately the same density of individuals over time. The EC values can be used to set technically defensible levels of protection, with the appropriate effect level being determined on the basis of data- and site specific considerations and dose-response relationships that are amenable for use as inputs in population models. Even without the use of predictive population models, the ECO10 or EC20 are commonly used values in risk assessment or criteria development with the assumption of adequate protection of populations. In the Se example presented here, there is strong evidence that egg hatchability is the critical toxicity endpoint for birds based on dietary organic Se exposures and that mallards are a sensitive bird species. These factors support that the dietary Se EC10 derived by Ohlendorf (2003) is sufficiently low to not have any measurable effects on aquatic birds in the field. Further, effect levels below the EC10 are likely to be statistically indistinguishable from the controls in most situations (as it was for Se in this example), and, for Se and other naturally occurring elements, it is not unusual for lower EC values to approach or fall below background levels at a site. A determination as to whether higher EC values would also provide an adequate level of protection could be made by appropriate population modeling. PMID- 19130655 TI - Temporal changes in contaminated sediment: a neglected aspect of ecological risk assessment? PMID- 19130656 TI - Back to basics in aquatic toxicology. PMID- 19130657 TI - Is it OK to fool Mother Nature? PMID- 19130658 TI - Current perspective of venous thrombosis in the upper extremity: a rebuttal. PMID- 19130659 TI - No-nano sunscreens? PMID- 19130660 TI - Hypertension and diabetes--double trouble. Get checked for diabetes if you have high blood pressure. PMID- 19130661 TI - Balancing hope and reality in heart failure. PMID- 19130662 TI - Joint inflammation may point the finger at heart disease. Rheumatoid arthritis and lupus raise the risk of heart attack and stroke. PMID- 19130663 TI - When quitters are winners. Breaking the addiction to cigarettes is tough, but the payoff is worth it. PMID- 19130664 TI - Trial questions beta blockers for all before noncardiac surgery. PMID- 19130665 TI - Ask the doctor. I need to catheterize myself because of bladder problems. In April 2007, I had five stents put in my heart and started taking Plavix. Since then, I sometimes see a light tinge of blood in the catheter bag, though lately the blood flow has been more substantial. The book "Worst Pills, Best Pills" says Plavix should be used for only a limited time, but my internist tells me I should be on it for the rest of my life. I am 88 years old and take aspirin (325 mg) every day. How long will I need to take Plavix? PMID- 19130666 TI - Ask the doctor. I often get mail from companies like Life Line Screening about having tests to look for "hidden" heart risks. The events are usually held at a local church and cost and $130. Are these tests valid? Are they worth the money? PMID- 19130667 TI - The top 10 health stories of 2008. PMID- 19130668 TI - What's up...and what's down. Winners and losers in some recent studies. PMID- 19130669 TI - The no-name cancer. Non-Hodgkin's lymphoma may not have much name recognition, but it's common and--the good news--often treatable, if not curable. PMID- 19130670 TI - By the way, doctor. I have injury-related osteoarthritis in my right ankle. My orthopedic surgeon said, in essence, that it's ankle fusion or nothing for me- and that there are no means of surgically restoring cartilage. What do you think? PMID- 19130671 TI - [The model of dopaminergic regulation of involuntary movements]. PMID- 19130672 TI - [Progress in clinical otorhinolaryngology--upper airway inflammation and vaccine therapy]. PMID- 19130673 TI - Reduction of acyl enolates of alpha-substituted beta-keto esters by bakers' yeast. AB - Enol esters of 2-substituted-3-oxoalkanoates of (Z)-configuration were reduced by bakers' yeast to chiral 2-substituted-3-hydroxyalkanoates. The syn-selectivities of this reaction increased compared with those of the reduction of the corresponding racemic keto esters. The reaction may proceed via an asymmetric hydrolysis of the enol esters, followed by the reduction of the resulting carbonyl group. PMID- 19130674 TI - Relationship between the ligand structure of copper and the stability of superoxide dismutase. AB - The ligand structure of chelated copper in recombinant human Cu(2)Zn(2)SOD (r hSOD) and its metal-substituted SODs was examined. In r-hSOD the configuration of the copper binding site was axially symmetrical and that of metal-substituted SODs had a tendency to change towards a tetrahedral type compared with r-hSOD except for Cu(2)(VO)(2)SOD. Cu(2)(VO)(2)SOD was not tetrahedral and in this case the ligand atom of copper was inclined to convert from nitrogen towards oxygen. In Cu(2)E(2)SOD, Ag(2)Cu(2)SOD, and Cu(2)(VO)(2)SOD the specific activities were lower but in CU(2)Cu(2)SOD and Cu(2)Co(2)SOD they were nearly identical with that of r-hSOD. These results suggested the ligand structure of chelated copper was not related to the specific activity of SOD. However, these SODs were less stable than r-hSOD against heat treatment and denaturing reagent. Further they were less stable against attack by an inactivator (hydrogen peroxide) except for Cu(2)(VO)(2)SOD. In this case the decreased stability of these SODs was associated with the change of the ligand structure of copper from that of r-hSOD. These results suggested that the presence of zinc contributed highly to the stable formation of the ligand structure of copper and the enzymatic stability. PMID- 19130675 TI - Fluorometric determination of sulfite in wine by N-(9-acridinyl)maleimide. AB - A highly sensitive fluorometric method for determining sulfite in wine is reported. N-(9-Aciridinyl)maleimide (NAM) reacted with the sulfite in wine quantitatively and gave strong fluorescent derivatives. The determinations were conducted by batch and high-performance liquid chromatographic methods. Sulfite contents in wine by NAM methods and the conventional methods showed good accordance. The sample size could be reduced to less than 1/10,000 by the NAM methods. PMID- 19130676 TI - Isolation and properties of a chromosome-dependent KHR killer toxin in Saccharomyces cerevisiae. AB - A strain of the yeast Saccharomyces cerevisiae coding for KHR on the chromosome secreted a toxin that kills sensitive yeasts. The transformants of multicopy vectors carrying the KHR gene could secrete 3-4-fold the killer toxin of the donor strain. This toxic substance was purified 80-fold in specific activity from the culture filtrate by gel filtration and hydrophobic column chromatography. The purified toxin gave a single protein band with molecular mass of 20 kDa on SDS PAGE and had an isoelectric point of pH 5.3. The toxin had novel killer activity against Candida glabrata and S. cerevisiae, but did not affect bacteria, fungi, or other yeasts. PMID- 19130677 TI - [1935: a fishing season on the coast of Sirte. The agricultural-fishing village of Az-Zuwaytinah, a failed colonial experiment in Cyrene]. PMID- 19130678 TI - Asian migration: human rights, human wrongs, citizenship and multiculturalism. PMID- 19130679 TI - Can the Ethiopian change his skin? The Beta Israel (Ethiopian Jews) and racial discourse. PMID- 19130680 TI - Care for orphans in nineteenth century Warsaw. PMID- 19130682 TI - First amendment -- California Supreme Court holds that free exercise of religion does not give fertility doctors right to deny treatment to lesbians. -- North Coast Women's Care Medical Group, Inc. v. San Diego County Superior Court, 189 P.3d 959 (Cal. 2008). PMID- 19130681 TI - Child in wealthy Warsaw bourgeois family: second half of the 19th and the early 20th century. PMID- 19130683 TI - Journal of Health Politics, Policy and Law. Editor's note. PMID- 19130684 TI - The nurse's role in managing necrotizing fasciitis. AB - Necrotizing fasciitis (NF), also known as "flesh-eating disease," is a rapid acting, potentially deadly infection. This condition is devastating in its ability to spread through the body at an alarming rate. It can be caused by almost any type of injury that introduces microorganisms into the subcutaneous tissues. The initial assessment can be deceptive, so awareness is the key to recognition. This condition often goes unrecognized until it is too late or major disfigurement results. Vital nursing strategies for managing the care of patients with NF include early recognition, accurate assessment, and aggressive treatment. PMID- 19130685 TI - Perioperative care of the patient with tuberculosis. AB - Approximately 1.7 billion people are thought to be infected with Mycobacterium tuberculosis. The estimated mortality rate is 3 million people per year. Multidrug resistant-tuberculosis (MDR-TB) and extensively drug-resistant TB (XDR TB) are serious problems that threaten global TB control. Patients who are inadequately treated remain chronic carriers and can spread the disease to family members and their communities. Treatment recommendations for TB include directly observed treatment and monitoring the patient's response to medications. Perioperative nurses must protect the patient and other health care workers when a patient with TB undergoes a surgical procedure. PMID- 19130686 TI - Perioperative grand rounds: A near miss lesson: check the wristband. PMID- 19130687 TI - Cross cultural confusions: indigenous traditions, legal confrontations and ethnographic uncertainties. PMID- 19130688 TI - Decentralisation, participation and accountability in Sahelian forestry: legal instruments of political-administrative control. PMID- 19130689 TI - Dispute over the sex education of children and young people during the inter-war years. PMID- 19130690 TI - [Noble savage, good citizen: American Indians and Christian reform projects of the 17th-18th centuries]. PMID- 19130691 TI - Our legacy and our future. PMID- 19130692 TI - How do you predict the eventual amount of spacing or crowding that will occur in a child in the mixed dentition stage? PMID- 19130693 TI - The orthodontist gave my patient a rapid maxillary expander and then a functional appliance with a headgear component. The patient has achieved an outstanding result! What biological mechanism allows these appliances to work? PMID- 19130694 TI - Should I be concerned if a patient requiring orthodontic treatment has an allergy to nickel? PMID- 19130695 TI - Practising oral-systemic medicine: the need for interprofessional education. PMID- 19130697 TI - Editorial: Child Support. PMID- 19130696 TI - Editorial: The Kindest Cut? PMID- 19130698 TI - Development of integrated testing strategies for REACH: motivation, background and introduction. Preface. PMID- 19130700 TI - [Current status of anti-tuberculosis policies and future challenge (2) The framework for anti-tuberculosis policies]. PMID- 19130699 TI - [Exercises, physical activities and public health (8): Promotion of physical activities utilizing health communication]. PMID- 19130702 TI - Difference of opinion. AMA, AHA at odds on behavior standards. PMID- 19130704 TI - Fighting to survive in Tulsa. OSU medical center may be made a public trust. PMID- 19130703 TI - Debt for debt? Payment-in-kind bond options present new wrinkle. PMID- 19130705 TI - Oldie, but a goodie. With credit still in the deep freeze, time is right for a modern Hill-Burton Act. PMID- 19130706 TI - Measure your efforts. Exemption relies on proper charity-care estimates. PMID- 19130707 TI - Drawing high interest. Alternative financing deals, guidance from GPOs playing bigger role in tech purchases. PMID- 19130701 TI - [Regional health for the 21st century (2) The population approach and an asset model]. PMID- 19130708 TI - Survey stresses value of leadership development for healthcare organizations. PMID- 19130709 TI - Reinventing healthcare leadership. PMID- 19130710 TI - Preparing the leadership pipeline for 21st century healthcare. PMID- 19130712 TI - MGMA pushes for standardized insurance cards. PMID- 19130711 TI - The 3 tech questions. PMID- 19130713 TI - Tech the halls. PMID- 19130714 TI - Stretched by the RAC. PMID- 19130715 TI - Latest strategies in diagnosis and treatment of PTSD. PMID- 19130716 TI - Contingency fees can silence a frivolous claim. PMID- 19130717 TI - GPs can help their patients adjust to chronic illness. PMID- 19130718 TI - Reducing exacerbations in patients with COPD. PMID- 19130719 TI - Managing patients with difficult asthma. PMID- 19130720 TI - Rapid assessment vital in stroke and TIA. PMID- 19130721 TI - On the therapeutic value of oil and water. 1868. PMID- 19130722 TI - Eye conditions. Part 2. PMID- 19130723 TI - Should GPs give penicillin for suspected meningococcal septicaemia? PMID- 19130725 TI - [Patient with Crohn's disease: now she complains about a swollen knee]. PMID- 19130726 TI - [Discussion of influenza vaccination: is the usefulness for old people overvalued?]. PMID- 19130727 TI - [Diabetics need encouragement: do not place your patients in a dock!]. PMID- 19130729 TI - [Inguinal hernia: diagnosis and therapy]. PMID- 19130728 TI - [Diagnostics and therapy of chronic diarrhea in elderly people]. PMID- 19130730 TI - [Doping in mass sport--what the family doctor needs to know]. PMID- 19130731 TI - [Dermatoses in obesity: sick skin over the adipose tissue]. PMID- 19130732 TI - Ganglioglioma originating in the cerebellum with a large cyst--a case report and review of the literature. AB - Here we report a rare case of cerebellar ganglioglioma accompanied by a large cyst, and present a review of the reported 28 cases with cerebellar ganglioglioma. An otherwise healthy 46-year-old woman complained of gradual headache and truncal ataxia. MRI revealed a huge cystic lesion with a mural nodule in the left cerebellar hemisphere. The tumor was resected totally. Histologically, it was composed of neuronal and glial elements, and was accordingly diagnosed as ganglioglioma. PMID- 19130733 TI - Occipital pilomyxoid astrocytoma in a 14-year-old girl--case report. AB - Pilomyxoid astrocytoma is a recently described tumor. Its most typical morphological characteristics are an angiocentric astrocytic proliferation embedded in a myxoid background. The behavior seems to be unfavorable due to the reported high rate of local recurrence. The earlier studies indicated that pilomyxoid astrocytoma typically affects young children and arises in the hypothalamic/chiasmatic region. We report a case of a 14-year-old patient with a 6-year history of absence seizure. Magnetic resonance imaging showed a right occipital lesion of approximately 3 cm in diameter. The patient underwent the surgical procedure with gross total excision. Histologically, the tumor was mainly composed of a monomorphous population of bipolar elongated piloid cells radially arranged around thin-walled blood vessels in a prominent myxoid background. There were focal hemorrhagic foci but no bona fide evidence of tumor necrosis or mitoses. Rosenthal fibers and eosinophilic granular bodies were not observed. The postoperative course was uneventful. No adjuvant therapy was administered. The patient is alive and well at 18-month follow-up. The case presented provides evidence that pilomyxoid astrocytoma can occur at a later age and can arise in regions different from hypothalamic/chiasmatic. PMID- 19130734 TI - Detecting chromosomal alterations at 1p and 19q by FISH and DNA fragment analysis -a comparative study in human gliomas. AB - Histological classification of gliomas is important for treatment and as a prognostic predictor, but classification by histology alone can be a challenge. Molecular genetic investigations, in particular the combined loss of the short arm of chromosome 1 and the long arm of chromosome 19 (LOH1p/19q), has become a significant predictor of outcome in oligodendrogliomas. 1p/19q alterations can be investigated by fluorescence in situ hybridization (FISH), but controversies persist in the interpretation ofresults. Another technique is polymerase chain reaction (PCR) analysis using microsatellites as primers and capillary electrophoresis or southern blot as detection method. The objective of the present study was to compare the accuracy, reliability and feasibility of detecting chromosomal changes at 1p/19q with PCR microsatellite analysis and FISH in glial tumors in the clinical laboratory, where often only small formalin-fixed paraffin-embedded samples are available. Commercial DNA and normal cortex were used for comparison. The material comprised 41 glial tumors including 10 oligodendrogliomas (WHO Grades II and III, 5 each), 10 mixed oligoastrocytomas (WHO Grades II and III, 5 each), 10 astrocytomas (WHO Grades II and III, 5 each), and 11 glioblastomas (WHO Grade IV). Our data confirmed a correlation between FISH and LOH fragment analysis in classical oligodendrogliomas and in mixed oligoastrocytomas. Disparity was found among the glioblastomas, where fragment analysis showed 1p/19q loss in three cases, with no changes detected by FISH. The fragment analysis seems reliable and implementable for LOH 1p/19q investigation without patient-related control material. PMID- 19130735 TI - Reliability and reproducibility of PCR-based testing of O6-methylguanine-DNA methyltransferase gene (MGMT) promoter methylation status in formalin-fixed and paraffin-embedded neurosurgical biopsy specimens. AB - OBJECTIVE: To analyze the reliability and reproducibility of PCR-based testing of O6-methylguanine-DNA methyltransferase gene (MGMT) promoter methylation status in formalin-fixed and paraffin-embedded (FFPE) neurosurgical biopsy specimens. MATERIALS: We used 6 FFPE neurosurgical temporal lobe specimens of children and young adults with drug-resistant epilepsy. Histopathologically, all specimens showed CNS tissue with gliosis but no tumor tissue. METHODS: For MGMT promoter methylation analysis, we used methylation specific PCR (MGMT MSP). In all 6 tissue specimens, 4 repetitive runs of MGMT MSP were performed. RESULTS: We obtained conclusive results only in 13/24 (54.2%) MGMT MSP analyses. In 5/13 (38.5%) successful MSP runs, the results indicated presence of a methylated MGMT promoter. In only 1/6 specimens, MSP yielded consistent results in all 4 repetitive runs. CONCLUSIONS: In our hands, MGMT MSP shows poor reliability and reproducibility of test results on FFPE neurosurgical tissue specimens. A more reliable method for diagnostic MGMT promoter methylation testing needs to be identified and validated by systematic testing of intra- and interlaboratory reliability and reproducibility. Alternatively, methods of tissue fixation that do not impair DNA quality but at the same time warrant high quality histopathology could facilitate molecular diagnostics. PMID- 19130736 TI - Angiocentric glioma: presentation of two cases with dissimilar histology. AB - OBJECTIVE AND IMPORTANCE: Angiocentric glioma (AG) is a recently described tumor of the brain which was included as a distinct entity in the 2007 WHO classification. To date only 26 cases have been reported in the literature. We describe two additional cases of this possibly confusing lesion of the brain. Emphasis is put on variations in the histopathological picture. CLINICAL PRESENTATION: The patients (20- and 55-year-old males) presented with seizures and headaches, respectively. Imaging examination showed a small cortical subcortical tumor in each case. Both tumors were totally removed. MATERIAL AND METHODS: Paraffin blocks from the two cases were examined with classical histopathology stainings and immunohistochemistry for GFAP, vimentin, EMA, neurofilament protein, synaptophysin, S100 protein, CD31, CD34, FVIII, smooth muscle actin and Ki67. RESULTS: The tumor proliferation was restricted around small intraparenchymal vessels. Immunohistochemistry demonstrated positivity for glial and negativity for vascular or neuronal markers. The cell shape and arrangement was different in the two cases. CONCLUSIONS: AG is a peculiar tumor of uncertain histogenesis but with certain glial reactivity. Histopathology is variable but restricted, for unknown reasons, to perivascular areas. Apparently, a better prognosis than for other gliomas is distinctive. Further studies are needed in order to expand the information regarding the clinical behavior and therapeutic approach of this tumor type. PMID- 19130737 TI - A case of an elderly adult presenting with obstructive hydrocephalus secondary to a rare hemorrhagic suprasellar pilocytic astrocytoma. AB - In this report, we present a 65-year-old man who presented with signs and symptoms consistent with impending brain herniation. Emergent imaging revealed a hyperdense mass in the suprasellar region. Urgent surgery was performed and final pathology eventuated a pilocytic astrocytoma. Although rare cases of suprasellar pilocytic astrocytoma in children and adults have been reported, we report an interesting case of a hemorrhagic suprasellar pilocytic astrocytoma in an elderly adult (without prior anticoagulant use) causing impending brain herniation secondary to obstructive hydrocephalus. PMID- 19130738 TI - Aspergillus terreus brain abscess mimicking tumor progression in a patient with treated glioblastoma multiforme. AB - OBJECTIVE: To detail a case of Aspergillus terreus brain abscess in a patient undergoing treatment for malignant glioma. Central nervous system aspergillosis usually occurs in patients with hematopoietic neoplasms or post transplantation, not in those with solid tumors. Most systemic invasive mold infections are attributable to Aspergillus fumigatus or Aspergillus flavus. PATIENT AND METHODS: The patient had received external beam radiation, temozolomide chemotherapy, and high-dose steroids, and had lymphopenia, but not sustained neutropenia. She developed a brain mass that mimicked tumor progression by neuroimaging criteria; infection was not a consideration. RESULTS: Brain biopsy showed fungal cerebritis and cultures grew A. terreus, a variant being reported with greater frequency as a pathogen in patients with risk factors for aspergillosis. CONCLUSION: Brain tumor patients who receive steroids to control their peritumoral edema may be particularly susceptible to cerebral A. terreus infection, especially when they additionally develop the lymphopenia commonly associated with temozolomide. PMID- 19130739 TI - A patient with MV2 subtype of sporadic Creutzfeldt-Jakob disease and atypical clinical presentation. AB - We report the case of a 71-year-old woman with progressive dementia over the course of 4 years, characterized by prominent pyramidal signs and by the lack of ataxia and other cerebellar signs. Creutzfeldt-Jakob disease (CJD) was not suspected during the patient's life. Autopsy brain tissue showed severe spongiform encephalopathy with kuru-like, but not florid, plaques in neocortex and cerebellum. Massive synaptic diffuse and plaque-like PrP(Sc) deposition was found in the cerebral cortex, striatum, cerebellum and brainstem. Genetic analysis revealed no PRNP gene mutations and methionine/valine heterozygosity (MV) at codon 129. The pathogenic scrapie prion protein (PrP(Sc)) pattern detected by Western blot was Type 2. However, this pattern showed a single unglycosylated band in contrast to the doublet described for MV2 subtype of sCJD with kuru plaques. In summary, this is an autopsy case report of a particular presentation of MV2 subtype of sCJD. PMID- 19130740 TI - The G11778A LHON mutation does not enhance ethambutol cytotoxicity in a cybrid model. AB - Leber's hereditary optic neuropathy (LHON) is a maternally inherited mitochondrial disorder, leading to a selective loss of retinal ganglion cells (RGC) and degeneration of the optic nerve, which results in severe visual impairment or even blindness. The primary causes are point mutations of the mitochondrial DNA (mtDNA), associated with aminoacid exchanges in complex I of the electron transport chain (ETC), which are thought to disturb oxidative ATP generation in the mitochondria. The major side effect of the antibiotic ethambutol, commonly used in tuberculosis therapy, is a retinopathy, which may lead to selective RGC loss, if not detected in an early stage. Moreover, LHON was reported to be elicited by ethambutol in some mutation carriers. OBJECTIVE: The present study intended to measure a possible synergism between mitochondrial dysfunction, caused by the most common LHON mutation (G11778A) and caused by ethambutol, which may lead to a higher cytotoxicity of the drug in LHON cells. MATERIAL: An NT2/D1 teratoma-derived LHON cybrid line and the parental cells. METHOD: Determination of ethambutol toxicity in both lines, using a microtiter tetrazolium assay, luminometric measurement of ATP/ADP ratios and determination of mtDNA copy numbers by Real-time PCR. RESULTS: Short-term ethambutol toxicity occurred only at micromolar concentrations, far beyond the estimated plasma peak concentrations of patients under antibiotic therapy. No significant difference occurred between both cell lines. The ATP/ADP ratios in the cybrids were surprisingly low, but showed no correlation with the mutational status of drug treated cells. The mtDNA copy number of treated LHON and parental cells did not differ significantly. CONCLUSIONS: Ethambutol shows no synergism with the most common primary LHON mutation with respect to mitochondrial energy production or mtDNA replication in cybrid cells, although the issue of ATP decline should be further addressed in neuronally differentiated cybrids with complete OXPHOS dependency. PMID- 19130741 TI - Altered distribution of lamin and emerin in muscle nuclei of sIBM patients. AB - OBJECTIVE: Sporadic inclusion body myositis (sIBM) is a chronic acquired inflammatory myopathy. The cause of sIBM remains unknown and its pathogenesis is controversial. There is a hypothesis [Karpati and Carpenter 1993] that the rimmed vacuoles result from nuclear breakdown, and IBM filaments are formed from components of the nuclear matrix. MATERIAL AND METHODS: For nuclear membrane protein detection, six IBM patients were studied using immunohistochemical and immunochemical techniques. RESULTS: It was demonstrated that in the interior of 10-15% myonuclei emerin and lamin A/C inclusions appeared constantly. This finding indicated an abberant localization of lamin A/C epitopes, the presence of presumptive lamin A (67 KDu) and emerin as in the affected nuclei. CONCLUSION: We support the suggestion that truncated, changed lamin A protein takes part in nuclear disintegration and rimmed vacuole formation. PMID- 19130742 TI - Dynamin 2-related centronuclear myopathy: clinical, histological and genetic aspects of further patients and review of the literature. AB - Centronuclear myopathy (CNM) is a slowly progressive congenital myopathy with characteristic histopathological findings of chains of centrally located myonuclei in a large number of muscle fibers. Recently, different missense mutations in the dynamin 2 gene (DNM2, 19p13.2) have been shown to cause autosomal dominant CNM. We re-evaluated patients with a histopathological diagnosis of CNM and report on the clinical phenotype, the biopsy findings and the genetic results of these patients and review the current literature. Two of the three patients showed an unusually late disease onset (> 40 years). Interestingly, intramuscular nerve fascicles found in the muscle biopsy of a patient harboring the E368K DNM2 mutation contained nerve fibers with disproportionately thin myelin sheaths. Schwann cells of unmyelinated nerve fibers showed abnormal plasma membrane and basal lamina protrusions, indicating peripheral nerve involvement. PMID- 19130743 TI - 125th anniversary of the Institute of Neurology (Obersteiner Institute) in Vienna. "Germ Cell" of interdisciplinary neuroscience. PMID- 19130744 TI - Sixtieth anniversary of the Institute of Medical Microbiology, Semmelweis University, Budapest (1948-2008). PMID- 19130745 TI - "Up-dating the monograph." [corrected] Cytolytic immune lymphocytes in the armamentarium of the human host. AB - The author of the monograph "Cytolytic Immune Lymphocytes..." (published in 2008 by Schenk Buchverlag Campus Dialog, Budapest, Passau, Pecs) proposed several research projects and described certain clinical events that require further elaboration and documentation. In this article the author provides what is required and has since become available. The first subject matter in question concerns the fusogenic viruses. The ancient fusogenic viruses might have created the first eukaryotic cell(s) by uniting archaeabacterial and prokaryotic/protobacterial protospheroplasts. Extant fusogenic viruses either produce tumor cell syncytia and lyse them, thus practicing viral oncolysis. Or, create chimaeric fusion products, the so-called "natural hybridomas", of lymphoma cells exhibiting transmembrane budding of retrovirus particles or envelope proteins, and anti-viral specific antibody-producing plasma cells. The second topic concerns the horizontal-lateral mode of acquisition of those genes, which were "present in the waiting" in the amphioxus, sea urchin, and the agnathans, and met in the primitive gnatostomata sharks to encode in unison the entire adaptive immune system. The consensus of opinion is such that these genes derived from newly acquired transposons/retrotransposons. The author points out that the extant Epstein-Barr virus harbors genes displaying sequence homology with those genes from the sharks up to mammals that regulate the somatic hypermutation of specific antibody production. The author proposes that an ancient herpesvirus might have propagated the V(D)J and RAG genes from sea urchins to sharks. The third area is that of lymphocytes cytotoxic/cytolytic to virally infected or malignantly transformed host cells. This discovery led to the adoptive immune lymphocyte therapy of tumors. Installed in the adaptive immune system are regulatory T cells and myeloid-derived suppressor cells for he protection of "self". Tumor cells masquerading as "self" are protected by these cells from attacks launched by immune T cells. The author supports the replacement of IL-2 by IL-15, inasmuch as IL-2 stimulates not only immune T cells, but also regulatory T cells expressing the CD25 IL-2 receptor. The administration of low dose whole body radiotherapy prior to immune lymphocyte therapy increases the efficacy of immune lymphocyte therapy. The author observed this phenomenon in the mid-1960s. The explanation of this phenomenon revealed itself just recently. In pre-irradiated hosts the intestinal wall becomes permeable to the gut flora; the intestinal bacteria activate the entire innate immune system in the mesenteric lymph nodes and a rapid activation of the adaptive immune faculties follows. PMID- 19130746 TI - Emergence of antiretroviral drug resistance in therapy-naive HIV infected patients in Hungary. AB - Mutations in the HIV-1 genes associated with resistance to antiretroviral drugs were detected also in primary HIV infected individuals who did not receive antiretroviral treatment. Drug resistance genotyping of HIV pol gene was done by in situ DNA hybridization using a Line Probe Assay and by direct sequencing. Viral variants harbouring resistance mutations such as: M41, T69R, K70R, M184V, T215Y in the pol gene were detected in 14% of the subjects. HIV mutants resistant to NRT inhibitors were found in 10 and 20% of patients infected before and after the year 2000, respectively. Multiple drug resistant viruses (2-3 drug classes) were present in 3.5% of the mainly recently infected patients. In protease gene only minor resistant mutations were found such as L101 and A71V. These findings indicate the evolution of drug resistance showing a correlation with the time of introduction of combination therapy in our country, where more than 70% of HIV infections were by homo/bisexual transmission. This confirms the transmission of drug-resistant HIV shown by genotype testing during primary infection in therapy naive patients and initiates serious clinical and public health consequences. PMID- 19130747 TI - Molecular characterisation, antibiotic sensitivity and serotyping of hungarian Streptococcus pneumoniae isolates. AB - In this study we summarise the results of the first major epidemiological survey of Streptococcus pneumoniae isolates from Hungary. Neither the incidence nor the level of resistance to penicillin of the strains was high, we observed, however, 40% macrolide resistance. The main underlying mechanism was the presence of the erm(B) gene, but we could find also strains with the mef gene, some of them with unusually high resistance levels. The main serogroups in ranking order were 6, 9, 19, 23 and 14, accounting together for 88.2% of all typed strains. We found a close correlation between serotypes and resistance. As a result of genotyping the penicillin non-susceptible strains, we found that the penicillin resistant isolates all belonged to the same PFGE clone, while in the intermediate group the diversity was greater. We could compare our isolates to the successful international PMEN clones, and proved the presence of some of them in Hungary. In general, we could establish the similarity of Hungarian pneumococci and other international isolates. PMID- 19130748 TI - Vancomycin resistant enterococci (VRE) still persist in slaughtered poultry in hungary 8 years after the ban on avoparcin. AB - In this report we examined the glycopeptide susceptibility of enterococci, isolated in 2005, from slaughtered animals, within the confines of Hungarian Antibiotic Resistance Monitoring System. We determined the presence of the van genes as well as their genetic relatedness in enterococci from poultry. Enterococcus sp. strains (n=175) were collected from intestinal samples of slaughtered poultry in 2005. The origin of the samples was registered at county level. After screening the strains with 30 mg vancomycin disc 19 (86%) intermediate resistant and 4 (3%) fully resistant strains were found. The distribution of minimum inhibitory concentration (MIC)-values among 23 enterococcus strains which were intermediate or resistant to vancomycin were 0.25 mg/L (4.4%), 2 mg/L (8.6%), 4 mg/L (8.6%), 8 mg/L (61%), 16 mg/L (8.6%) and 256 mg/L (8.6%). The MICs of teicoplanin were 0.25 mg/L (4.3%), 1 (8.6%), 4 mg/L (78.3%), 16 mg/L (4.3%) and 256 mg/L (4.3%). The two most vancomycin-resistant strains were vanA carriers (1 E. faecalis and 1 E. faeciuum). The farms that produced these strains can be reservoirs of VRE and the affected farms should change the technology of disinfection and breeding in order to prevent the emergence of high numbers of human VRE isolates in Hungary. PMID- 19130749 TI - Comparison of Streptococcus mutans strains from children with caries-active, caries-free and gingivitis clinical diagnosis by pulsed-field gel electrophoresis. AB - A study was conducted to compare the DNA structure of Streptococcus mutans strains in children with caries-active, caries-free, and gingivitis clinical diagnosis. Twenty-eight Streptococcus mutans strains from 100 children's plaques were examined by pulsed-field gel electrophoresis (PFGE) method. The classified strains were closely related to one another, though the strains originated from different disease groups. Three identical pairs were found, but the pairs in two cases belonged to different disease groups. The results of the PFGE experiments suggest that there is no correlation between the different DNA patterns ofS. mutans strains and their cariogenecity. So the different DNA strains ofS. mutans are not the only determining factor in the development of dental caries. PMID- 19130750 TI - Seroprevalence of HSV-2 in Hungary and comparison of the HSV-2 prevalence of pregnant and infertile women. AB - Herpes simplex virus type 2 infection is a quite common but frequently asymptomatic, therefore undiagnosed condition. Genital HSV-2 infection may cause neonatal herpes, enhances HIV transmission and may play a role in infertility. To evaluate the prevalence of HSV-2 in Hungary we tested 2500 serum samples for the presence of anti-HSV-2 IgG by ELISA method. According to our results Hungary belongs to the low-infected countries, the HSV-2 seroprevalence grows with age and is significantly higher among women than in men. We also examined the serostatus of 512 pregnant women and 539 women attending infertility clinics. Results show that the HSV-2 prevalence is significantly higher among women attending infertility clinics and the seropositivity of pregnant women is similar to that of the general Hungarian women population with the same age. PMID- 19130751 TI - Expression patterns of cel5A-cel5B, two endoglucanase encoding genes of Thermobifida fusca. AB - Expression patterns of cel5A and cel5B, two endoglucanase encoding genes of Thermobifida fusca were compared by quantitative real-time PCR. With Avicel as carbon source the transcript level of cel5A continuously increased until the 10th hour of incubation and then a sharp decrease was observed, whereas cel5B presented a slow constitutive expression on this substrate. When the microcrystalline cellulose powder MN300 was used as the inducing carbon source, the expression patterns of the two genes were similar. A low initial level of expression was followed by a rapid increase at the 5th hour of incubation; a transient repression was then observed at the 10th hour but after this sampling time, the expression levels started to increase again. The relative expression levels of cel5A were always higher than those of cel5B. Differences in transcription patterns of these two genes can be explained with the imperfect structure of the CelR binding regulatory region of cel5B. PMID- 19130752 TI - Characterisation of the first VIM metallo-beta-lactamase-producing Pseudomonas aeruginosa clinical isolate in Serbia. AB - From the Central-East European region the first VIM metallo-beta-lactamase (MBL) producing Pseudomonas aeruginosa strains were published from Croatia, Poland and Hungary. The aim of this study was to assess the contribution of MBL-production to carbapenem-resistance among P. aeruginosa clinical isolates in the Military Medical Academy (MMA) in Belgrade, Serbia between August 2004 and September 2007. Only one P. aeruginosa isolate with strain number 722 proved MBL-positive that harboured a novel class 1 integron with a bla(VIM-2)-like cassette in the first position, followed by orfD, a putative gene with unknown function. Our data indicate that MBL-producing strains occur at a prevalence of less than 1% among imipenem-nonsusceptible P. aeruginosa clinical isolates in this Belgrade hospital. The newly identified VIM MBL-producing P. aeruginosa strain 722 could be assigned to serotype O11, and it was panresistant to all antimicrobials tested. The isolate displayed sequence type ST235 by multilocus sequence typing which is the founder sequence type of the previously identified international clonal complex CC11 that already contains bla(VIM)-positive isolates from Italy, Greece, Sweden, Hungary and Poland. In conclusion, this is the first report of VIM MBL-producing P. aeruginosa from Serbia and also of the occurrence of such isolates belonging to the international clonal complex CC11 in this country. PMID- 19130753 TI - Observations on environmental factors in connection with avian botulism outbreaks in a Hungarian wetland habitat. AB - In this study, the relationship between five water-quality parameters (water temperature, pH, conductivity, water-soluble oxygen (WSO) and organic matter content (chemical oxygen demand-COD)) and avian botulism cases were examined using statistical methods. Five-five sampling points, located in high and low avian botulism risk (HR and LR) areas of Lake 'Kis-Balaton' were chosen, respectively. Data from 5 HR and 5 LR years were processed. In some cases, significant (P<0.05) differences were discovered in water temperature, pH, WSO and COD values between HR and LR sites and years. Discriminant analysis verified positive relationship between the occurrence of avian botulism outbreaks and characteristics of the sampling sites in 16/20 cases (80.0%). In the remaining instances, the role of other, not examined factors could be also important. PMID- 19130754 TI - America's best health plans. PMID- 19130755 TI - Ten tips for picking a health plan. PMID- 19130757 TI - Thanks, we'd rather not. PMID- 19130756 TI - Doing battle with your plan. PMID- 19130758 TI - Behind the rankings. PMID- 19130759 TI - Best Health Plans PT-Directory. Commercial plans. PMID- 19130760 TI - Best Health Plans PT-Directory. Medicaid plans. PMID- 19130761 TI - A better way to screen for cardiovascular disease? PMID- 19130762 TI - Visual search of emotional faces. Eye-movement assessment of component processes. AB - In a visual search task using photographs of real faces, a target emotional face was presented in an array of six neutral faces. Eye movements were monitored to assess attentional orienting and detection efficiency. Target faces with happy, surprised, and disgusted expressions were: (a) responded to more quickly and accurately, (b) localized and fixated earlier, and (c) detected as different faster and with fewer fixations, in comparison with fearful, angry, and sad target faces. This reveals a happy, surprised, and disgusted-face advantage in visual search, with earlier attentional orienting and more efficient detection. The pattern of findings remained equivalent across upright and inverted presentation conditions, which suggests that the search advantage involves processing of featural rather than configural information. Detection responses occurred generally after having fixated the target, which implies that detection of all facial expressions is post- rather than preattentional. PMID- 19130763 TI - The recall of information from working memory. Insights from behavioural and chronometric perspectives. AB - We describe and evaluate a recall reconstruction hypothesis for working memory (WM), according to which items can be recovered from multiple memory representations. Across four experiments, participants recalled memoranda that were either integrated with or independent of the sentence content. We found consistently longer pauses accompanying the correct recall of integrated compared with independent words, supporting the argument that sentence memory could scaffold the access of target items. Integrated words were also more likely to be recalled correctly, dependent on the details of the task. Experiment 1 investigated the chronometry of spoken recall for word span and reading span, with participants completing an unfinished sentence in the latter case. Experiments 2 and 3 confirm recall time differences without using word generation requirements, while Experiment 4 used an item and order response choice paradigm with nonspoken responses. Data emphasise the value of recall timing in constraining theories of WM functioning. PMID- 19130764 TI - Transposed-letter priming effects for close versus distant transpositions. AB - Transposing two internal letters of a word produces a perceptually similar item (e.g., CHOLOCATE being processed as CHOCOLATE). To determine the precise nature of the encoding of letter position within a word, we examined the effect of the number of intervening letters in transposed-letter effects with a masked priming procedure. In Experiment 1, letter transposition could involve adjacent letters (chocloate-CHOCOLATE) and nonadjacent letters with two intervening letters (choaolcte-CHOCOLATE). Results showed that the magnitude of the transposed-letter priming effect--relative to the appropriate control condition--was greater when the transposition involved adjacent letters than when it involved nonadjacent letters. In Experiment 2, we included a letter transposition condition using nonadjacent letters with one intervening letter (cholocate-CHOCOLATE). Results showed that the transposed-letter priming effect was of the same size for nonadjacent transpositions that involved one or two intervening letters. In addition, transposed-letter priming effects were smaller in the two nonadjacent conditions than in the adjacent condition. We examine the implications of these findings for models of visual-word recognition. PMID- 19130765 TI - Recognizing users of the recognition heuristic. AB - The recognition heuristic is hypothesized to be a frugal inference strategy assuming that inferences are based on the recognition cue alone. This assumption, however, has been questioned by existing research. At the same time most studies rely on the proportion of choices consistent with the heuristic as a measure of its use which may not be fully appropriate. In this study, we propose an index to identify true users of the heuristic contrasting them to decision makers who incorporate further knowledge beyond recognition. The properties and the applicability of the proposed index are investigated in the reanalyses of four published experiments and corroborated by a new study drawn up to rectify the shortcomings of the reanalyzed experiments. Applying the proposed index to explore the influence of knowledge we found that participants who were more knowledgeable made use of the information available to them and achieved the highest proportion of correct inferences. PMID- 19130766 TI - Controlled processing in single-prime negative priming. AB - Negative priming refers to delayed responses to previously ignored distractors. Unlike conventional studies of negative priming in which the attentional selection of a target against its distractors is required in prime trials (prime selection negative priming), in single-prime negative priming, a prime stimulus is presented briefly. To further investigate the nature of single-prime negative priming, its properties were examined. In Experiment 1, the proportion of repetition was varied. The effect of single-prime negative priming was reduced when the proportion of repetition was high. In addition, Experiment 2 showed that high memory load could hamper the single-prime negative priming effect. Overall, the current study indicates controlled processing in single-prime negative priming and similarities between single-prime negative priming and prime selection negative priming. PMID- 19130767 TI - Influence of intermixed emotion-relevant trials on the affective Simon effect. AB - "Good" and "bad" vocal responses are faster when an irrelevant emotional stimulus feature corresponds with the response than when it does not, a phenomenon known as the affective Simon effect. Two experiments investigated how this effect was influenced by an intermixed emotion-relevant evaluation task. In Experiment 1, four schematic faces (friendly, happy, hostile, sad) were used for the affective Simon task and four different images (bird, heart, gun, ghost) for the evaluation task, whereas in Experiment 2 the schematic faces were used for both tasks. Mixed compatible emotion-relevant trials increased the affective Simon effect in both experiments, but mixed-incompatible emotion-relevant trials did not influence it. Also, the advantage of the compatible mapping over the incompatible mapping increased in mixed conditions rather than decreased. These results differ from those obtained when visual-manual tasks for which location is relevant and irrelevant are mixed. They confirm that enhancement of the affective Simon effect when the Simon task is mixed with a compatible emotion-relevant task is due to increased salience of the affective valence. PMID- 19130768 TI - [Primary skin lesions in the mouth. 5 years later erosions of the vulva]. PMID- 19130769 TI - [Sequelae of the JUPITER study. What is changing in primary prevention? (interview by Dr. med. Dirk Einecke)]. PMID- 19130771 TI - [DGIM against mail orders of prescription drugs. Risk of adulterated drugs]. PMID- 19130770 TI - [What is the status of the medical case consultation in group practice?]. PMID- 19130772 TI - [Novel therapies in oncology: how about adverse effects?]. PMID- 19130773 TI - ['How to convince people with type 2 diabetes to start with insulin? Treatment options for patients with psychological insulin resistance']. PMID- 19130774 TI - When medicine meets marketing. PMID- 19130775 TI - The happiness effect. How emotions and even behaviors can spread like an epidemic. PMID- 19130776 TI - Permanent birth control. Skip the scalpel. Two minutes, two inserts--and your childbearing years are over. PMID- 19130777 TI - ERs in distress: ACEP. Report cites states for not enough support, resources. PMID- 19130778 TI - Payment disclosure push. Experts say device exemption clouds Mass. proposal. PMID- 19130779 TI - Scrutiny on bonds. Public repository to hold annual data. PMID- 19130780 TI - Exception to the rule. Summa to control costs, charity care with physicians. PMID- 19130781 TI - Spirit of Excellence Award for Community--winner. Finger on community's pulse. PMID- 19130782 TI - Spirit of Excellence Award for Quality--honorable mention. Positive aggression. PMID- 19130783 TI - Becoming 'biocitizens'. High-tech tools help, but who'll interpret the data? PMID- 19130784 TI - Families after death, families after birth. PMID- 19130785 TI - The last will and testament in literature: rupture, rivalry, and sometimes rapprochement from Middlemarch to Lemony Snicket. AB - Although the psychological literature on the last will and testament is sparse, authors of fiction and memoir have filled the gap, writing in rich detail about the impact of wills on families. Henry James, George Eliot, J. R. Ackerley, and others reveal that a will is not only a legal document but a microcosm of family life: a coded and non-negotiable message from the will's writer to its intended readers, the heirs, delivered at a stressful time and driving home the truth that options for discussion between testator and heirs are now gone, all factors which may intensify the ambivalence of grief and stall its resolution. Among the problems the authors chronicle: reinvigorated sibling rivalries, vindictive testators, and the revelation of traumatic family secrets. Writers also demonstrate how contemporary social factors, such as divorce, second families, and geographic distance between family members, may complicate wills and ensuing family relations. Exemplary wills, or will-like documents, appear in fiction by Maria Katzenbach and Marilynne Robinson, allowing the living to make rapprochements with the dead, and pointing to testamentary strategies clinicians might develop to lead to a resolution of grief. The depth of these writers' accounts allows clinicians to imagine points at which they might productively intervene in matters pertaining to a will. PMID- 19130786 TI - Thoughts on the importance of wills in family relationships: a clinical response to Elizabeth Stone. PMID- 19130787 TI - New evidence for the social embeddedness of infants' early triangular capacities. AB - Infants appear to be active participants in complex interactional sequences with their parents far earlier than previously theorized. In this report, we document the capacity of 3-month-old infants to share attention with two partners (mothers and fathers) simultaneously, and trace links between this capacity and early family group-level dynamics. During comprehensive evaluations of the family's emergent coparenting alliance completed in 113 homes, we charted infants' eye gaze patterns during two different mother-father-infant assessment paradigms. Triangular capacities (operationalized as the frequency of rapid multishift gaze transitions between parents during interactions) were stable across interaction context. Infants exhibiting more advanced triangular capacities belonged to families showing evidence of better coparental adjustment. Theoretical and practice implications of these findings are discussed. PMID- 19130788 TI - Synchrony in the triad: a microlevel process model of coparenting and parent child interactions. AB - Guided by a microanalytic approach to the study of relationships, we assessed parent, infant, and coparental behaviors during triadic interactions in 94 parents and their 5-month-old firstborn child. Relational behaviors in each family subsystem--mother-infant, father-infant, and coparenting--were microcoded. Marital satisfaction and infant temperament were self-reported. No differences were found in the infants' behavior toward mother and father or in the time spent with each parent. Mothers' and fathers' relational behavior during parent-infant episodes were generally comparable, yet mothers vocalized more and the latency to father's displaying positive affect was longer. Conditional probabilities indicated that under conditions of coparental mutuality, fathers showed more positive behaviors than mothers. Lag-sequential analysis demonstrated that change in the infant's social focus between parents followed change in coparental behavior. Fathers' coparental mutuality was independently predicted by maternal behavior during mother-child episodes, father marital satisfaction, and infant difficult temperament, whereas mothers' coparental mutuality was only linked with fathers' relational behavior. Results highlight the importance of including a microlevel perspective on the family system at the first stages of family development. PMID- 19130790 TI - Parent characteristics as antecedents of maternal gatekeeping and fathering behavior. AB - The present study examined the role of prebirth parent characteristics as predictors of maternal gatekeeping (mothers' attempts to encourage or discourage fathers' interaction with their infant) and fathering behavior. Parents' idealization of their relationships within their families of origin, beliefs about the roles of fathers, and personality attributes (negative emotionality and communion) were assessed before their infant's birth. At 3.5 months postpartum, maternal gatekeeping behaviors (negative control, facilitation) and fathers' involvement and competence with their infants were assessed during observation of triadic play and child care. Results suggest reciprocal relations between maternal gatekeeping and fathering behavior. Furthermore, greater paternal communion was associated with greater paternal competence during play, whereas greater maternal communion was associated with lower paternal competence during child care. Greater maternal communion and greater maternal idealization related to fathers' lower relative involvement during play. As for maternal gatekeeping behavior, high negative emotionality in 1 parent was only accompanied by high levels of inhibitory maternal gatekeeping when the other parent had less progressive beliefs about the father's role. The implications of these findings for clinicians and practitioners are discussed. PMID- 19130789 TI - Withdrawal from coparenting interactions during early infancy. AB - This study examines early withdrawal in the coparenting system, and the utility of a brief problem-solving discussion about coparenting responsibilities as a means for evaluating such withdrawal. One hundred and fifteen couples were evaluated both prenatally and at 3 months postpartum. During prenatal assessments, parents rated their personalities and completed marital assessments. After the baby arrived, they completed a negotiation task in which they discussed disputes about parenting roles and responsibilities, and interacted together with the baby in a triadic play assessment. Fathers' but not mothers' withdrawal during coparenting negotiations was associated with greater disengagement and less warmth during triadic play and with fathers' feelings that mothers did not respect their parenting. Fathers' but not mothers' withdrawal during coparenting negotiations was also forecast by low ego resilience and by an increase in depressive symptomatology during the postpartum. As the negotiation task appeared to be an effective provocateur of withdrawal when confronting coparenting disagreement, it may prove useful for eliciting this aspect of coparental process in work with couples. PMID- 19130791 TI - Gaining ground from a family and cultural legacy: a teen mother's story of repairing the world. AB - A multigenerational longitudinal study of teen mothering provided prospective data on the intergenerational continuities and discontinuities in parenting traditions and caregiving legacies. Families that included a teen mother were first interviewed intensively in 1988-1989 and were reinterviewed in 1993, 1997, 2001, and 2005. All studies in the series were based on the phenomenology of everyday practices and the assumption that the self is constituted by practical, historical, and embodied understandings. Data were analyzed using the hermeneutic approach. PMID- 19130792 TI - Individual resiliency factors from a genetic perspective: results from a twin study. AB - This article is part of the Twin Mother's Study, a study that examines influences on maternal adjustment. A number of studies have investigated the importance of genetic factors for mental health, but few of these examine how genes and the environment influence resiliency/salutogenic factors. This article investigates the relative importance of genetic and environmental influences on resiliency/salutogenic factors. This study includes 326 twin pairs (150 monozygotic and 176 dizygotic) who are mothers, who are living with their spouse, and who are part of the Swedish twin register. Using self-report structured questionnaires, we assessed salutogenic factors, depression, and quality of life; however, we analyzed the questionnaires completed by the mothers. Statistical analyses were conducted using structural equation modeling. We conclude that nonshared environmental components were of principal importance in individual resiliency/salutogenic factors in a genetically informative design, but we also noted that genetic influences were important. The shared environment had mainly no effect. PMID- 19130793 TI - Family secrets and family functioning: the case of donor assistance. AB - The purpose of this study was to examine the relationship between adult offspring's perception of family functioning and of parental use of topic avoidance to maintain secrecy regarding the use of donor assistance to conceive. A cross-sectional design was used to study a convenience sample of 69 young adult donor offspring who completed a demographic questionnaire, a topic avoidance scale relative to each of their rearing parents, and the Beavers Self Report Family Instrument. Findings indicated that participants perceived both parents as avoiding the topic of donor assistance more than other topics, mothers as avoiding all topics less than fathers, and topic avoidance was negatively associated with family functioning. Mothers' general topic avoidance was the strongest predictor of family functioning. Parents' disclosing together was predictive of higher family functioning. Implications for practice and future research are suggested. PMID- 19130794 TI - Russian Old Believers: genetic consequences of their persecution and exile, as shown by mitochondrial DNA evidence. AB - In 1653, the Patriarch Nikon modified liturgical practices to bring the Russian Orthodox Church in line with those of the Eastern (Greek) Orthodox Church, from which it had split 200 years earlier. The Old Believers (staroveri) rejected these changes and continued to worship using the earlier practices. These actions resulted in their persecution by the Russian Orthodox Church, which forced them into exile across Siberia. Given their history, we investigate whether populations of Old Believers have diverged genetically from other Slavic populations as a result of their isolation. We also examine whether the three Old Believer populations analyzed in this study are part of a single gene pool (founder population) or are instead derived from heterogeneous sources. As part of this analysis, we survey the mitochondrial DNAs (mtDNAs) of 189 Russian Old Believer individuals from three populations in Siberia and 201 ethnic Russians from different parts of Siberia for phylogenetically informative mutations in the coding and noncoding regions. Our results indicate that the Old Believers have not significantly diverged genetically from other Slavic populations over the 200 300 years of their isolation in Siberia. However, they do show some unique patterns of mtDNA variation relative to other Slavic groups, such as a high frequency of subhaplogroup U4, a surprisingly low frequency of haplogroup H, and low frequencies of the rare East Eurasian subhaplogroup D5. PMID- 19130795 TI - Genetic characterization and assessment of authenticity of ancient Korean skeletal remains. AB - To study the maternal lineage history of Korea, we extracted DNA from the skeletal remains of 35 museum samples (some dating back to the Paleolithic Age) excavated from 11 local burial sites scattered throughout southern Korea. Mitochondrial DNA (mtDNA) control region sequences (HV1, HV2, and HV3) were successfully determined for 11 samples with no sharing of the control region polymorphisms with individuals involved in the laboratory analyses. Each of the 11 mtDNAs was assigned to the appropriate East Asian mtDNA haplogroup according to the haplogroup-specific control region mutation motif and diagnostic coding region single nucleotide polymorphism. The successful mtDNA haplogroup determination for each ancient Korean mtDNA and the confirmation of the absence of abnormal mutations based on the haplogroup-directed database comparisons indicates that there is no mosaic structure from cross-contamination or sample mix-up or other errors in our mtDNA sequences. The presence of haplogroups B, D, and G in the prehistoric age is consistent with the hypothesis that the early Korean population has a common origin in the northern regions of the Altai Mountains and Lake Baikal of southeastern Siberia. In addition, the modern Korean population, which possesses lineages from both southern and northern haplogroups, suggests additional gene flow from southern Asian haplogroups in recent times, but many more ancient samples need to be analyzed to directly tell whether there was regional continuity or replacement of early lineages by other lineages in ancient Korea. PMID- 19130796 TI - Genetic affinities of the Siddis of South India: an emigrant population of East Africa. AB - Historical records indicate that the Portuguese brought the African Siddis to Goa, India, as slaves about 500 years ago. Subsequently, the Siddis moved into the interior regions of the state of Karnataka, India, and have remained there ever since. Over time the Siddis have experienced considerable cultural changes because of their proximity to neighboring population groups. To understand the biological consequences of these changes, we studied the Siddis to determine the extent of genetic variation and the contributions from the African, European, and Indian ancestral populations. In the present study we typed the Siddis for 20 polymorphic serological, red cell, and Alu insertion-deletion loci. The overall pattern of phenotype (and genotype) distribution is in accordance with Hardy Weinberg expectations. Considering the ethnohistorical records and the availability of secondary-source genetic data, we used two data sets in the analysis: one comprising eight serological and red cell enzyme markers with eight population groups and another comprising six Alu insertion-deletion markers with seven tribal groups of South India. The dendrograms generated from these two data sets on the basis of genetic distance analysis between the selected populations of African, European, and Indian descent reveals that the Siddis are closer to the Africans than they are to the South Indian populations. Genetic admixture analysis using a dihybrid model (19 loci) and a trihybrid model (10 loci and 8 loci) shows that the predominant influence comes from the Africans, a lesser contribution from the South Indians, and a slight contribution from the Portuguese. Thus the original composition of the African genes among the Siddis has been diluted to some extent by the contribution from southern Indian population groups. There is no nonrandom association of alleles among a set of 10 genetic marker systems considered in the present study. The demonstration of genetic homogeneity of the Siddis, despite their admixed origin, suggests the utility of this population for genetic and epidemiological studies. PMID- 19130797 TI - Effect of sociocultural cleavage on genetic differentiation: a study from North India. AB - Indian populations possess an exclusive genetic profile primarily due to the many migratory events, which caused an extensive range of genetic diversity, and also due to stringent and austere sociocultural barriers that structure these populations into different endogamous groups. In the present study we attempt to explore the genetic relationships between various endogamous North Indian populations and to determine the effect of stringent social regulations on their gene pool. Twenty STR markers were genotyped in 1,800 random North Indians from 9 endogamous populations belonging to upper-caste and middle-caste Hindus and Muslims. All nine populations had high allelic diversity (176 alleles) and average observed heterozygosity (0.742 +/- 0.06), suggesting strong intrapopulation diversity. The average F(ST) value over all loci was as low as 0.0084. However, within-group F(ST) and genetic distance analysis showed that populations of the same group were genetically closer to each other. The genetic distance of Muslims from middle castes (F(ST) = 0.0090; DA = 0.0266) was significantly higher than that of Muslims from upper castes (F(ST) = 0.0050; DA = 0.0148). Phylogenetic trees (neighbor-joining and maximum-likelihood) show the basal cluster pattern of three clusters corresponding to Muslims, upper-caste, and middle-caste populations, with Muslims clustered with upper-caste populations. Based on the results, we conclude that the extensive gene flow through a series of migrations and invasions has created an enormous amount of genetic diversity. The interpopulation differences are minimal but have a definite pattern, in which populations of different socioreligious groups have more genetic similarity within the same group and are genetically more distant from populations of other groups. Finally, North Indian Muslims show a differential genetic relationship with upper- and middle-caste populations. PMID- 19130798 TI - Y-chromosome variability in four Native American populations from Panama. AB - The allele and haplotype frequencies for 13 Y-chromosome short tandem repeats (STRs) [nine STR loci of the minimal Y-chromosome haplotype (DYS19 - DYS385a - DYS385b - DYS389I - DYS389II - DYS390 - DYS391- DYS392 - DYS393) plus four additional loci (DYS388, DYS426, DYS439, DXYS156)] were determined in 99 males from 4 Panamanian native American populations, including the Chibcha-speaking Ngobe and Kuna and the Choco-speaking Embera and Wounan. Fifty haplotypes were identified, of which 48 (96%) were specific to a single population and 29 (63%) were found in only a single individual. Gene diversity per locus per population ranged from 0 to 0.814, with the highest gene diversity present at the DYS389II locus in the Embera. The haplotypic discrimination capacity was low, ranging from 42.3% in the Kuna to 63.1% in the Wounan. The four tribes showed a high degree of differentiation both at the Y chromosome and in the mitochondrial genome, highlighting the importance of genetic structure even in geographically proximate and linguistically related populations. PMID- 19130799 TI - Angiotensin converting enzyme gene polymorphism (insertion/deletion) and hypertension in adult Asian Indians: a population-based study from Calcutta, India. AB - The angiotensin converting enzyme (ACE) gene insertion/deletion polymorphism has been identified as a potential genetic risk factor for essential hypertension. The purpose of the present study is to investigate the association of the insertion/deletion polymorphism of the ACE gene with essential hypertension in adult Asian Indians. Three hundred fifty (184 males and 166 females) adult (30 years and older) Asian Indians of Calcutta and its suburbs participated in this population-based cross-sectional study. Anthropometric measures, lipids profiles, blood glucose, and blood pressure measures were collected from participants. ACE insertion/deletion polymorphism was determined by agarose gel electrophoresis and D/D typing was further reconfirmed using insertion-allele-specific amplification. Essential hypertension was defined as a systolic blood pressure (SBP) greater than 160 mm Hg and/or a diastolic blood pressure (DBP) greater than 90 mm Hg or use of any antihypertensive treatment by participants. Significantly higher SBP, DBP, and mean arterial pressure were recorded in D/D subjects compared to I/I and I/D subjects. We also observed that the odds of being hypertensive were 7.483 (95% CI = 1.746, 30.192) in D/D individuals compared to those carrying one or no D alleles. This finding suggests that ACE insertion/deletion polymorphism is associated with essential hypertension in Asian Indians. Moreover, individuals who are homozygous for the D allele of the ACE gene are more likely to have essential hypertension. PMID- 19130800 TI - Comparison of Korean and Japanese head and face anthropometric characteristics. AB - The primary purpose of this research is to collate, compare, and discuss the presently available data for head and face dimensions among Korean and Japanese ethnic groups. Classification of Korean male and female head and face types is simpler than classification of Japanese subjects. Male groups have more statistically significant morphological differences than females, and Japanese subjects display larger values for head and face measurement categories than Korean subjects. Japanese item values for head and face dimensions show distinct differences between male and female subjects compared to Korean subjects. Japanese male subjects have distinct differences from Korean male subjects and relatively lower values for head and face dimensions as age increases compared to Korean male subjects. Generally, female subjects have no regular tendency according to age compared to male subjects. PMID- 19130801 TI - The mystery of SARS. PMID- 19130803 TI - How progress makes us sick. PMID- 19130802 TI - Economies on empty. PMID- 19130804 TI - A genuinely healthy debate. PMID- 19130805 TI - A struggle inside AA. PMID- 19130806 TI - Lollipops & lawsuits. PMID- 19130807 TI - To treat the dead. PMID- 19130808 TI - Just say no--to bad science. PMID- 19130809 TI - The time is all in your mind. PMID- 19130810 TI - Brutal case studies. PMID- 19130811 TI - The dieter's dilemma. PMID- 19130812 TI - What the Beatles gave science. PMID- 19130814 TI - Logging on to lose those extra pounds. PMID- 19130813 TI - His fine feathered friends, and ours. PMID- 19130815 TI - These poles are made for walking. PMID- 19130816 TI - Look, ma, I'm going meatless! PMID- 19130817 TI - More information, please. PMID- 19130818 TI - Inside Karen's crowded mind. PMID- 19130819 TI - Caution: killing germs may be hazardous to your health. PMID- 19130820 TI - Thanks, but no thanks. PMID- 19130821 TI - When baby catches a cold. PMID- 19130822 TI - The kids aren't all right. PMID- 19130823 TI - How to mend a sick system. PMID- 19130824 TI - Family leave under fire? PMID- 19130825 TI - This is no way to cure cancer. PMID- 19130826 TI - Stronger, faster, smarter. PMID- 19130827 TI - Exercise is a state of mind. PMID- 19130828 TI - On your mark... PMID- 19130829 TI - An 'exercise snack' plan. PMID- 19130830 TI - What the doctors do. PMID- 19130831 TI - Cancer and Staying fit. PMID- 19130832 TI - HDL drugs: what now? PMID- 19130833 TI - How to help your heart. PMID- 19130834 TI - How to pay for old-age care. PMID- 19130836 TI - Trouble in toyland. PMID- 19130835 TI - Looking for answers to my nephew's death. PMID- 19130837 TI - Beware the blinkers. PMID- 19130838 TI - Know thyself--man, rat or bot. PMID- 19130839 TI - Accidents will happen. PMID- 19130840 TI - Attack of the diet cokes. PMID- 19130841 TI - Let's get physical. PMID- 19130842 TI - Knees in need of help. PMID- 19130843 TI - Running on empty. PMID- 19130844 TI - Stopping a kid killer. PMID- 19130845 TI - (Rethinking) gender. PMID- 19130846 TI - The truths we wants to deny. PMID- 19130847 TI - Health planning. PMID- 19130848 TI - Failing the health test. PMID- 19130849 TI - What kills one women every minute of ever day? PMID- 19130850 TI - When does your brain stop making new neurons? PMID- 19130851 TI - How to think like a scientist. PMID- 19130852 TI - Let's not hide health costs. PMID- 19130853 TI - Fundamentalism in science. PMID- 19130854 TI - Dietary spices in health and diseases: I. AB - Spices are heterogeneous collections of a wide variety of volatile and non volatile staple dietary additives. India with its wide climatic conditions and topographical features naturally possesses wide variety of medicinal flora. Spices have a diverse array of natural phytochemicals that have complementary and overlapping actions, including antioxidant effects, modulation of detoxification enzymes, stimulation of immune system, reduction of inflammation, modulation of steroid metabolism and antibacterial and antiviral effects. In the present essay, various studies on effects of different well characterized spices on molecular, cellular, autocrine, paracrine and endocrine mechanisms and their role in neuromodulation, immunoinodulation, anti inflammatory, antioxidant, anti carcinogenic, antimutagenic and psychoactive phenomena have been reviewed. PMID- 19130855 TI - Effect of yoga based lifestyle intervention on subjective well-being. AB - Yoga is assuming importance in improving mental health and quality of life in the treatment of a number of psychiatric and psychosomatic disorders. The present study was a prospective controlled study to explore the short-term impact of a comprehensive but brief lifestyle intervention, based on yoga, on subjective well being levels in normal and diseased subjects. Normal healthy individuals and subjects having hypertension, coronary artery disease, diabetes mellitus or a variety of other illnesses were included in the study. The outcome measures were 'subjective well being inventory' (SUBI) scores, taken on the first and last day of the course. The inventory consists of questions related to one's feelings and attitude about various areas of life, such as happiness, achievement and interpersonal relationship. There was significant improvement in the subjective well being scores of the 77 subjects within a period of 10 days as compared to controls. These observations suggest that a short lifestyle modification and stress management educational program leads to remarkable improvement in the subjective well being scores of the subjects and can therefore make an appreciable contribution to primary prevention as well as management of lifestyle diseases. PMID- 19130856 TI - Effect of exogenous selenium on nicotine induced hyperlipidemia in rats. AB - The effect of two different doses (1 microg Se/Kg and 50 microg Se/Kg Body wt) of selenium on nicotine induced hyperlipidemia was investigated in rats. Results revealed that nicotine intake caused an increase in concentration of cholesterol, triglycerides, free fatty acids, phospholipids and low density lipoprotein compared to control group. Coadministration of selenium along with nicotine reduced the levels of lipids compared to nicotine group. This reduction was due to reduction in the biosynthesis of lipids as evidenced by the reduced activity of HMGCoA reductase and lipogenic enzymes. Nicotine intake also reduced the absorption of selenium in the intestine. Histopathological studies revealed that selenium at a dose of 1 microg was more effective in reducing lipid levels and higher dose of selenium was toxic. PMID- 19130857 TI - Effects of Nigella sativa supplementation for one month on cardiac reserve in rats. AB - Nigella sativa (N. sativa) has a long history of use in folk medicine. In a current study performed in this laboratory, two-month dietary supplementation with N. sativa extract to normal rats has shown a homogenous cardiac hypertrophy and enhanced cardiac contractility at baseline conditions. In the present study, shorter (one-month) duration of oral N. sativa administration was adopted to detect possible earlier cardiac responses. In addition, in vitro cardiac stress by the beta adrenoceptor agonist isoproterenol was used to assess the intrinsic cardiac reserve mechanisms. The hearts of Nigella-treated rats developed a moderate but significant hypertrophy that was evident by an increase in the heart weight to body weight ratio. The observed Nigella-induced cardiac hypertrophy was associated with an increase in the baseline cardiac inotropic properties as well as the maximal peak tension generation upon progressive cardiac stress by isoproterenol infusion. The demonstrated selective enhancement of the inotropic reserve favours the physiological nature of Nigella-induced cardiac hypertrophy, similar to that provoked by exercise training. PMID- 19130858 TI - Antioxidants and lipid peroxidation in gestational diabetes--a preliminary study. AB - Increased free radical activity in gestational diabetes (GDM) can lead to a host of damaging and degenerative maternal and fetal complications. Hence antioxidant levels in blood of GDM mothers and cord blood were estimated. Erythrocyte glutathione (GSH), superoxide dismutase (SOD) and thiobarbituric acid reactive substances (TBARS), plasma vitamins C and E and serum total glutathione-S transferase (GST), protein thiols and ceruloplasmin (Cp) were estimated spectrophotometrically in maternal blood of age matched controls and mothers with GDM and also in cord blood samples of the above. There was a significant increase in the erythrocytic GSH, serum total GST and protein thiols in GDM maternal blood when compared to controls whereas erythrocytic SOD exhibited a marked decrease in GDM cases. The changes in plasma vitamins C and E, Cp and erythrocytic TBARS in GDM were not significantly different from controls. Cord blood levels of protein thiols were also significantly increased in GDM. No significant changes were observed in the serum Cp and GST levels of the same. Hence, elevated glucose levels can induce oxidative stress in GDM mothers. PMID- 19130859 TI - Ptychodiscus brevis toxin decreases the spontaneous activity of rat right atria involving muscarinic receptors and potassium channels. AB - Marine dinoflagellate Ptychodiscus brevis toxin (PbTx), is known to produce toxic effects on cardiovascular system. The present experiments were conducted to evaluate the effect of synthetic phosphorus containing Ptychodiscus brevis toxin on spontaneously beating right atrium in vitro. The PbTx (0.84-84 microM) decreased the rate and force of right atrial contractions in a concentration dependent manner. Ethanol, a vehicle present in highest concentration of PbTx, had no effect on atrial rate or force of contraction. Pretreatment with atropine blocked the PbTx-induced decrease in atrial rate and force of contraction. The tetraethylammonium, a potassium channel blocker, blocked the PbTx-induced decrease in atrial rate and force, where as, L-type of calcium channel blocker, nifedipine blocked the PbTx-induced force of contraction but not the rate changes. The results indicate that the PbTx decreased the atrial rate and force of contraction via cholinergic receptors involving K+ channel. PMID- 19130860 TI - Effect of six weeks yoga training on weight loss following step test, respiratory pressures, handgrip strength and handgrip endurance in young healthy subjects. AB - The present study was designed to test whether yoga training of six weeks duration modulates sweating response to dynamic exercise and improves respiratory pressures, handgrip strength and handgrip endurance. Out of 46 healthy subjects (30 males and 16 females, aged 17-20 yr), 23 motivated subjects (15 male and 8 female) were given yoga training and the remaining 23 subjects served as controls. Weight loss following Harvard step test (an index of sweat loss), maximum inspiratory pressure, maximum expiratory pressure, 40 mm endurance, handgrip strength and handgrip endurance were determined before and after the six week study period. In the yoga group, weight loss in response to Harvard step test was 64 +/- 30 g after yoga training as compared to 161 +/- 133 g before the training and the difference was significant (n = 15 male subjects, P < 0.0001). In contrast, weight loss following step test was not significantly different in the control group at the end of the study period. Yoga training produced a marked increase in respiratory pressures and endurance in 40 mm Hg test in both male and female subjects (P < 0.05 for all comparisons). In conclusion, the present study demonstrates attenuation of the sweating response to step test by yoga training. Further, yoga training for a short period of six weeks can produce significant improvements in respiratory muscle strength and endurance. PMID- 19130862 TI - Effect of extract of Benincasa hispida on oxidative stress in rats with indomethacin induced gastric ulcers. AB - This study was undertaken to determine the healing of ulcers induced by indomethacin due to antioxidant role of fruit extract of Benincasa hispida (Ashgourd) on ulcers in rats. Malondialdehyde (MDA) in RBC and antral homogenate was determined to measure tissue oxidation. Superoxide dismutase (SOD) in RBC and antral homogenate, plasma and homogenate vitamin C were estimated as measures of antioxidant defense. On induction of gastric ulcer, there was significant increase in SOD in RBC and homogenate levels and vitamin C in plasma. There was an apparent decrease in ulcer index in animals treated with fruit extract. There was significant decrease in MDA with concomitant decrease in SOD and vitamin C levels in the treated rats when compared to those not treated with fruit extract. Benincasa hispida has been shown to contain certain active principles like terpenes, flavanoid C--glycosides and sterols which have antioxidant effects. These probably inhibit gastric mucosal injury by scavenging the free radicals and repress production of SOD and vitamin C in these rats. PMID- 19130863 TI - A comparative study of median nerve somatosensory evoked potential in the totally blind and normal subjects. AB - The totally blind individuals are often considered to be compensated for their visual loss by augmentation in the auditory and tactile perceptions as against the normal sighted individuals. The objective of the present work was to study the somatosensory evoked potentials (SEPs) after median nerve stimulation in the wrist of totally blind and normal sighted individuals. SEPs were recorded after median nerve stimulation in the wrist of 15 Braille reading totally blind females and compared with 15 age matched normal sighted females. Latency and amplitudes of SEPs (N9, N13, and N20) were measured and analyzed statistically. Amplitude of N20 SEP was significantly larger in the totally blind than in normal sighted individuals (P < 0.05). The amplitudes of N9 and N13 SEPs and the latencies of all SEPs showed no significant differences. The congenitally blind individuals have larger N20 amplitude, suggestive of greater somatosensory cortical representation than normal sighted individuals. PMID- 19130861 TI - Attenuation of the effect of lindane on immune responses and oxidative stress by Ocimum sanctum seed oil (OSSO) in rats. AB - Present study was done to evaluate the effect of Ocimum sanctum seed oil (OSSO) on the immunotoxicity and oxidative activity of lindane in rats. Rats were divided into four groups (n = 8) and were treated with lindane (10 mg/kg, po) and/or OSSO (1 mg/kg, po) during the study period. Humoral immunity was assessed by measuring haemagglutination titre to sheep red blood cells (SRBC) and delayed type hypersensitivity (DTH) was assessed by measuring foot pad thickness. Lindane showed significant decrease in anti-SRBC antibody titre and also decreased percentage change in foot pad thickness in DTH response as compared to control group. OSSO per se produced significant increase in anti-SRBC antibody titre, but did not produce significant change in the foot pad thickness as compared to control group. However, it significantly antagonized the effect of lindane on the anti-SRBC antibody titre and foot pad thickness parameters. Lindane produced oxidative stress as indicated by increase in the levels of MDA and decrease in GSH levels. Treatment with OSSO per se showed antioxidant activity and also reversed the oxidative stress produced by lindane. The results suggest that OSSO can attenuate the immunotoxicity and oxidative stress produced by lindane. PMID- 19130864 TI - Sensory nerve conduction velocity is greater in left handed persons. AB - Dependence of nerve conduction velocity on factors like age, sex and temperature is well known. Effect of handedness on nerve conduction velocity has not been studied adequately. In this study correlation between handedness and motor as well as sensory conduction velocity of right as well as left median nerve was studied in 84 medical students in the age group between 17-21 years out of which 72 were right handed and 12 were left handed. In addition auditory and visual reaction times were also studied. Sensory conduction velocity in both right and left median nerve was significantly higher in left handers as compared with right handers whereas, the motor conduction velocity in right handers and left handers was not different. The auditory and visual reaction times were also not different in these groups. Because of greater sensory conduction velocity in left handers there is a requirement of different set of normal reference values for left handers in clinical diagnosis. PMID- 19130865 TI - Comparative study of pulmonary function variables in relation to type of smoking. AB - This study was intended to compare pulmonary function parameters in beedi smokers, cigarette smokers and subjects who smoked both beedi and cigarette on the basis of the dynamic ventilatory pulmonary function tests. The pulmonary function tests were done on 188 subjects which included, 48 beedi smokers, 48 cigarette smokers, 43 others who smoked both beedi as well as cigarette and 49 non-smokers (control). In beedi smokers the values of Forced Expiratory Volume in 1st second (FEV1), Peak Expiratory Flow Rate (PEFR), Forced Expiratory Flow (FEF) 25%-75%, FEV1/FVC%, BMI and weight were significantly lower (P < 0.01) than those of cigarette smokers and non-smokers. Moreover, the Forced Vital Capacity (FVC) and Maximal Voluntary Ventilation (MVV) of beedi smokers were significantly (P < 0.0001) lower than those of non-smokers. It may be concluded that pulmonary functions are more affected in beedi smokers than in cigarette smokers. PMID- 19130866 TI - Immediate effects of right and left nostril breathing on verbal and spatial scores. AB - The immediate effect of two yoga breathing techniques was assessed on verbal and spatial memory tasks, considered hemisphere-specific. Forty-five participants (24 males; age range 20 to 45 years (mean age 27.1 +/- 8.1 years) were randomly allocated to three groups (n = 15 each) and were assessed immediately before and after 45 minutes of three breathing practices i.e., right nostril yoga breathing, left nostril yoga breathing, or breath awareness as a control intervention. Spatial memory scores increased after left nostril yoga breathing compared to before (by 16 percent, P = 0.03, paired t-test). Hence, breathing through the left nostril increased performance in a spatial cognitive task, corresponding to the cerebral hemisphere contralateral to the patent nostril. PMID- 19130867 TI - Blood leukocyte profile in different phases of menstrual cycle. AB - The optimal availability of immune cells in the peripheral blood stream of women plays a critical role in their response to disease and therapeutic interventions. Interaction between the reproductive and immune system plays an important immunoregulatory role. This study was designed to examine the impact of different phases of menstrual cycle on the blood leukocytes. Twenty-four healthy women in their reproductive age group and having regular menstrual cycle were studied during menstrual, proliferative and secretary phases of menstrual cycle. Total leukocyte count, absolute and differential counts of neutrophils, lymphocytes and mixed cells (includes eosinophils, basophils and monocytes) were analyzed. Results showed that the variations in the different types of leukocytes during different phases of menstrual cycle were not statistically significant. No significant inter group difference, except for the significant decrease in differential lymphocyte percentage in proliferative phase as compared to menses were observed. PMID- 19130868 TI - Atherogenic lipid risk factors in men classified as overweight and obese according to the preliminary WHO guidelines for Asians. PMID- 19130869 TI - Are fresh water fish safe to eat? PMID- 19130871 TI - Practicing another form of preventive medicine. PMID- 19130870 TI - Transport of glucose in isolated goat ileum preparation: an alternative research tool for absorption study. PMID- 19130872 TI - March of Dimes releases premature birth report card: Kentucky receives "F''--KMA joins healthy babies coalition. PMID- 19130873 TI - Drive cancer out: a physician-led anti-smoking program directed at teens and adolescents. AB - Kentucky is among the states with the highest smoking-related mortality and youth smoking rates. Drive Cancer Out is a physician-led program that assesses and attempts to influence health literacy among Kentucky adolescents. Surveys on fifth-grade students identify social risk factors for smoking initiation and propose methods to decrease the rate of smoking among teenagers. PMID- 19130874 TI - Money, happiness, and suffering. PMID- 19130876 TI - 50 ways to improve your life: your body. PMID- 19130875 TI - Soldiering on. Military parents battle the system to help their autistic kids. PMID- 19130877 TI - 50 ways to improve your life: your mind. PMID- 19130878 TI - Unrelated developmental neurotoxicants elicit similar transcriptional profiles for effects on neurotrophic factors and their receptors in an in vitro model. AB - Diverse developmental neurotoxicants can often produce similar functional and behavioral outcomes. We examined an organophosphate pesticide (diazinon), an organochlorine pesticide (dieldrin) and a metal (Ni(2+)) for effects on the expression of neurotrophic factors and their receptors and modulators in differentiating PC12 cells, an in vitro model of neuronal development. Each agent was introduced at 30 microM for 24 or 72 h, treatments devoid of cytotoxicity. Using microarrays, we examined the mRNAs encoding members of the fibroblast growth factor (fgf) family, the neurotrophins (ntfs), brain-derived neurotrophic factor (bdnf), nerve growth factor (ngf), the wnt and fzd gene families, and the receptors and modulators for each class. All three agents evoked highly concordant patterns of effects on genes encoding the fgf family, whereas the correlations were poor for the group comprising bdnf, ngf and their respective receptors. For wnt, fzd and their receptors/modulators, the relationships between diazinon and dieldrin were highly concordant, whereas the effect of Ni(2+) was less similar, albeit still significantly correlated with the others. Our results show that otherwise disparate developmental neurotoxicants converge on common sets of neurotrophic pathways known to control neuronal differentiation, likely contributing to similarities in functional outcomes. Further, cell culture models can provide a useful initial screen to identify members of a given class of compounds that may be greater or lesser risks for developmental neurotoxicity, or to provide an indication of agents in different classes that might produce similar effects. PMID- 19130880 TI - Kinetic solubility and dissolution velocity of rutin nanocrystals. AB - Lyophilized rutin nanocrystals were intensively evaluated regarding their physicochemical properties with respect to particle size analyses, crystallinity, kinetic solubility and dissolution behavior. The particle size was determined by photon correlation spectroscopy (PCS) and laser diffraction (LD). DSC and X-ray diffraction were used to study the crystalline state of rutin nanocrystals. In a period of 1 week, the kinetic solubility was determined using a shaker at 25 degrees C. DSC and X-ray diffraction analyses showed that lyophilized rutin nanocrystals prepared by high pressure homogenization remained in crystalline state. Lyophilized rutin nanocrystals could be re-dispersed completely in water and the kinetic solubility in water increased to 133 microg/ml.. Lyophilized rutin nanocrystals were almost completely dissolved within 15 min in water, buffer of pH 1.2 and buffer of pH 6.8. In contrast, only 70% of rutin raw material (rutin microcrystals) was dissolved within 15 min. The superior physicochemical properties of rutin nanocrystals should overcome the absorption problem in the gastrointestinal tract and increase the bioavailability. PMID- 19130881 TI - Characterisation of density distributions in roller-compacted ribbons using micro indentation and X-ray micro-computed tomography. AB - Roller compaction is one stage in a dry granulation process to produce free flowing granules. Its proper understanding is essential in optimising manufacturing efficiency and product quality. Roller compaction produces a compacted strip or "ribbon", which is then milled to produce granules. For a given milling condition, the density distribution in the ribbons determines the properties of the granules (particularly their size distribution and strength). Therefore, knowing the density distributions in the ribbons is very important in improving the effectiveness of the roller compaction process and the quality of the granules produced. In this paper, the density distribution in roller compacted ribbons of microcrystalline cellulose (Avicel PH102) has been examined using three different techniques: (1) sectioning; (2) micro-indentation and (3) X ray micro-computed tomography. It has been shown that with proper calibration all three techniques can essentially produce the same results, but with a different degree of resolution (scale of scrutiny). In addition, the influence of process conditions, such as roll gap, roll speed and the presence or absence of lubrication, on the ribbon density distributions has also been investigated. Flow into the press is often constrained by the presence of "cheek plates", which prevent lateral powder movement. In this type of arrangement, it is found that non-uniform powder feeding occurs in the compaction region, induced by the friction between the powder and the cheek plates; as a result, the densities in the middle of the ribbon width are generally higher than those close to the edges. It has also been shown that higher average ribbon densities are obtained when the roll gap, roll speed, or the friction between the powder and the side cheek plates is reduced. PMID- 19130879 TI - Leptin signaling in brain: A link between nutrition and cognition? AB - Leptin is a protein hormone that acts within the hypothalamus to suppress food intake and decrease body adiposity, but it is increasingly clear that the hypothalamus is not the only site of leptin action, nor food intake the only biological effect of leptin. Instead, leptin is a pleiotropic hormone that impinges on many brain areas, and in doing so alters food intake, motivation, learning, memory, cognitive function, neuroprotection, reproduction, growth, metabolism, energy expenditure, and more. This diversity of function also means that a dysregulation of leptin secretion and signaling can have far reaching effects. To date research on leptin signaling has focused primarily on the hypothalamus, and the result is a relative lack of information regarding the impact of leptin signaling and leptin resistance in non-hypothalamic areas, despite a growing literature implicating leptin in the regulation of neuronal structure and function in the hippocampus, cortex and other brain areas associated with cognition. PMID- 19130882 TI - Global gene expression profiling of progesterone receptor modulators in T47D cells provides a new classification system. AB - Progesterone receptor modulators (PRMs) play an important role in women's health. They are widely used in oral contraception or hormone therapy, and provide an attractive treatment approach for gynecological disorders such as uterine leiomyomas, endometriosis or breast cancer. Due to the broad range of activities, various studies were conducted to assess progesterone receptor antagonists (PAs) and selective progesterone receptor modulators (SPRMs) with respect to progesterone receptor (PR) agonistic and antagonistic activities in vivo. These properties are not always adequately reflected in classical in vitro models, especially differences in the agonistic potential of SPRMs, such as asoprisnil, J1042, and J912, and mixed antagonists, such as mifepristone, are not sufficiently substantiated. The effects of PRMs upon gene expression in progesterone target tissues such as breast epithelium and uterus are poorly understood. This study compares the properties of PR ligands using mammalian two hybrid assays and gene expression profiling. The protein-protein interaction analyses in HeLa cells provide for specific ligand-induced PR conformations, whereas Affymetrix GeneChip HG-U133Plus2.0 analyses in T47D breast cancer cells indicate the transcriptional activity on the level of target genes. The analyses comprise the pure agonist R5020, the non-steroidal PR modulator PRA-910, SPRMs (J1042, asoprisnil, J912), the mixed antagonist mifepristone, classical antagonists (onapristone, ZK 137316) and the pure antagonist lonaprisan to consider all types of ligands described before. Marginal differences were identified in coactivator interaction profiles at all, but significant differences between SPRMs and PR antagonists (PAs) were observed in recruiting the LXXLL-motif containing peptide (LX-H10), very similar to in vivo activities in endometrial transformation in the rabbit (McPhail test). Global gene expression profiles demonstrated progesterone-independent effects for all PR modulators examined and emphasised similarities of asoprisnil and J1042 compared to J912 and all types of PR antagonists. In summary, the data support the popular concept of PR modulator classification in agonists, selective progesterone receptor modulators, mixed and pure antagonists. It further refines previous classification models and accentuates unique effects for each PR modulator. PMID- 19130883 TI - DHA and cholesterol containing diets influence Alzheimer-like pathology, cognition and cerebral vasculature in APPswe/PS1dE9 mice. AB - Cholesterol and docosahexenoic acid (DHA) may affect degenerative processes in Alzheimer's Disease (AD) by influencing Abeta metabolism indirectly via the vasculature. We investigated whether DHA-enriched diets or cholesterol-containing Typical Western Diets (TWD) alter behavior and cognition, cerebral hemodynamics (relative cerebral blood volume (rCBV)) and Abeta deposition in 8- and 15-month old APP(swe)/PS1(dE9) mice. In addition we investigated whether changes in rCBV precede changes in Abeta deposition or vice versa. Mice were fed regular rodent chow, a TWD-, or a DHA-containing diet. Behavior, learning and memory were investigated, and rCBV was measured using contrast-enhanced MRI. The Abeta load was visualized immunohistochemically. We demonstrate that DHA altered rCBV in 8 month-old APP/PS1 and wild type mice[AU1]. In 15-month-old APP/PS1 mice DHA supplementation improved spatial memory, decreased Abeta deposition and slightly increased rCBV, indicating that a DHA-enriched diet can diminish AD-like pathology. In contrast, TWD diets decreased rCBV in 15-month-old mice. The present data indicate that long-term dietary interventions change AD-like pathology in APP/PS1 mice. Additionally, effects of the tested diets on vascular parameters were observed before effects on Abeta load were noted. These data underline the importance of vascular factors in the APP/PS1 mouse model of AD pathology. PMID- 19130884 TI - Paradoxical delay in the onset of disease caused by super-long CAG repeat expansions in R6/2 mice. AB - Huntington's disease (HD) is caused by an expanded CAG repeat in the HD gene. The pathological threshold for expansion in HD is around 36 CAG repeats, although 'super-long' expansions are found in brains of HD patients. We examined the effect of varying the CAG repeat length (from 170 to 450) on behavior and neuropathology of R6/2 mice. Unexpectedly, we found that increasing the repeat length delayed onset of disease and prolonged survival, from around 4 months to over 18 months in mice with the longest repeats. The delay in onset correlated with a delayed appearance of neuronal intranuclear inclusions (NIIs). However, super-long CAG repeats are not neuroprotective. Mice carrying 2 copies of the mutant transgene die earlier than those carrying a single copy. Furthermore, neurodegeneration is present in super-long repeat length mice at mid-stage disease, whereas little neurodegeneration is seen in mice with shorter CAG repeats until end stage. Expanding the CAG repeat beyond the range where NII formation is the dominant pathology has unmasked a slowly progressing neurological phenotype in R6/2 mice with brain pathology, including the identification of a novel form of inclusion, that more closely resembles that seen in adult onset cases of HD. This mouse may represent a better model for adult-onset HD than R6/2 mice with shorter repeats. PMID- 19130885 TI - Cerebrospinal fluid antibodies to oxidized LDL are increased in Alzheimer's disease. AB - Lipoprotein oxidation may play an important role in the pathogenesis of Alzheimer's Disease (AD), and therefore, we investigated cerebrospinal fluid (CSF) antibodies to oxidized low-density lipoprotein (OxLDL) in patients with AD and other neurodegenerative dementias. IgM and IgG antibody titers to OxLDL were measured in 50 CSF samples and 11 plasma samples using chemiluminescent ELISA. All CSF samples contained IgG antibodies, and also most IgM, binding to OxLDL. CSF antibodies to OxLDL were not related to CSF protein or albumin concentrations or plasma antibodies to OxLDL. Competition immunoassay for specificity demonstrated that about 50% of the CSF IgG binding to OxLDL was inhibited by soluble OxLDL. CSF IgG antibodies to OxLDL were significantly increased in AD patients compared to controls and to patients with frontotemporal lobar degeneration. The role of these antibodies in CSF is unknown and further investigations are needed. PMID- 19130886 TI - Is Akt the "Warburg kinase"?-Akt-energy metabolism interactions and oncogenesis. AB - The serine/threonine kinase Akt - also known as protein kinase B (PKB) - has emerged as one of the most frequently activated protein kinases in human cancer. In fact, most, if not all, tumors ultimately find a way to activate this important kinase. As such, Akt activation constitutes a hallmark of most cancer cells, and such ubiquity presumably connotes important roles in tumor genesis and/or progression. Likewise, the hypermetabolic nature of cancer cells and their increased reliance on "aerobic glycolysis", as originally described by Otto Warburg and colleagues, are considered metabolic hallmarks of cancer cells. In this review, we address the specific contributions of Akt activation to the signature metabolic features of cancer cells, including the so-called "Warburg effect". PMID- 19130887 TI - Alternative splicing of the eag potassium channel gene in Drosophila generates a novel signal transduction scaffolding protein. AB - The Drosophila eag gene has been shown to regulate neuronal excitability, olfaction, associative learning and larval locomotion. Not all of the roles of this gene in these processes can be explained by its function as a voltage-gated potassium channel. In this study, we show that the eag gene is spliced in a PKA- and PKC-regulated manner to produce a protein lacking channel domains. This protein, in the context of activated PKA, can engage cellular signaling pathways that alter cell structure. Nuclear localization is necessary for C-terminal mediated effects, which also require MAPK. The requirement for PKA/PKC activation in the synthesis and function of this novel protein suggests that it may couple membrane events to nuclear signaling to regulate neuronal function on long time scales. PMID- 19130888 TI - Pain-associated signals, acidosis and lysophosphatidic acid, modulate the neuronal K(2P)2.1 channel. AB - Pain is a physiological state promoting protective responses to harmful episodes. However, pain can become pathophysiological and become a chronic disruptive condition, damaging quality of life. The mammalian K(2P)2.1 (KCNK2, TREK-1) channel, expressed in sensory neurons of the dorsal root ganglia, was previously identified as a polymodal molecular sensor involved in pain perception. Here, we report that two pain-associated signals, external acidosis and lysophosphatidic acid (LPA), known to rise during injury, inflammation and cancer, profoundly down modulate human K(2P)2.1 activity. The pH regulatory effect was mediated by activation of proton-sensitive G-protein coupled receptors and phospholipase C. Physiological concentrations of LPA overcame the effects of known K(2P)2.1 activators, such as arachidonic acid, lysophosphatidylcholine and temperature, by activating cell-surface receptors stimulating the G(q) pathway. Furthermore, we identified three K(2P)2.1 carboxy-terminal residues that mediate both pH and LPA regulatory effects. Our results highlight the important role of K(2P)2.1 channels as receptors for mediators known to cause nociception. PMID- 19130889 TI - Cloning, expression, isotope labeling, purification, and characterization of bovine antimicrobial peptide, lactophoricin in Escherichia coli. AB - Lactophoricin (LPcin-I) is a 23-amino acid peptide that corresponds to the carboxyterminal 113-135 region of component-3 of proteose peptone (PP3), a minor phosphoglycoprotein found in bovine milk. It has been reported that lactophoricin has antibacterial activity and a cationic amphipathic helical structure, but its shorter analogous peptide (LPcin-II), a 17-amino acid peptide, corresponding to the 119-135 region of PP3 does not display antibacterial activity. LPcin-I and LPcin-II have similar charge ratios and identical hydrophobic/hydrophilic sectors, according to their helical wheel projection patterns, and both peptides show cationic amphipathic helical folding and interact with membranes. However, it is known that only LPcin-I incorporates into planar lipidic bilayers to form voltage-dependent channels. In this study, the authors cloned and expressed the two recombinant peptides as ketosteroid isomerase (KSI) fusion proteins inclusion bodies in Escherichia coli. These peptides were subjected to NMR structural studies to explore their structure-activity relationships. Fusion proteins were purified by Ni-NTA affinity chromatography under denaturing conditions, and recombinant LPcin-I and LPcin-II were released from fusion by CNBr cleavage. Final purifications of LPcin-I and LPcin-II were achieved by preparative reversed phase high performance liquid chromatography. Using these methods, we obtained several tens of milligrams of uniformly and selectively (15)N labeled peptides per liter of growth, which was sufficient for solid-state NMR spectroscopy. Peptides were identified by tris-tricine polyacrylamide gel electrophoresis and HSQC spectra. Initial structural data were obtained by solution NMR spectroscopy and compared in membrane-like environments. PMID- 19130890 TI - Macrophage colony-stimulating factor (CSF-1) induces pro-inflammatory gene expression and enhances antimicrobial responses of goldfish (Carassius auratus L.) macrophages. AB - We report on the regulation of pro-inflammatory functions of goldfish macrophages and induction of gene expression by recombinant goldfish CSF-1 (rgCSF-1). Recombinant goldfish TNFalpha-2 (rg TNFalpha-2), rgIFNgamma but not rgTGFbeta induced time-dependent increase of CSF-1 expression in macrophages. Treatment of goldfish macrophages with rgCSF-1 increased expression of several immune genes including CXCL-8 (=IL-8), CCL-1, TNFalpha-1, TNFalpha-2, IL-1beta-1, IL-1beta-2, IL-12-p35, IL-12-p40, IFN, IL-10 and iNOS A and B. The rgCSF-1 treatment did not significantly alter the mRNA levels of TGFbeta and NRAMP in macrophages up to 48h post treatment. However, at 72h post treatment, the expression of TGFbeta increased whereas that of NRAMP decreased. The treatment of macrophages with rgCSF-1 enhanced their respiratory burst and nitric oxide responses that were abrogated after addition of soluble CSF-1 receptor (sCSF-1R) to cell cultures. Macrophages exhibited a concentration-dependent chemotactic response toward rgCSF 1 as well as an increase in phagocytic activity that was abrogated after addition of sCSF-1R to cell cultures. Our results indicate that in addition to being an important growth factor of goldfish macrophages, rgCSF-1 also plays a central role in the regulation of their pro-inflammatory responses. PMID- 19130891 TI - The relationships between daily life events and the availabilities of serotonin transporters and dopamine transporters in healthy volunteers--a dual-isotope SPECT study. AB - The aim of this study was to examine the relationships between the daily life events and the availabilities of serotonin transporters (SERT) and dopamine transporters (DAT) in healthy volunteers. Thirty-seven volunteers, 19 males and 18 females, were recruited. The SERT availability was approximated using SPECT and [123I] ADAM while the DAT availability was approximated using SPECT and [99mTc] TRODAT-1. All participants completed the Recent Life Change Questionnaire. The results showed that SERT and DAT may be sensitive to the presence of stressful events. PMID- 19130892 TI - Modified oxidosqualene cyclases in the formation of bioactive secondary metabolites: biosynthesis of the antitumor clavaric acid. AB - Hypholoma sublateritium is a basidiomycete that produces the antitumor compound clavaric acid. We cloned a gene encoding an oxidosqualene cyclase (occ) that is involved in the conversion of oxidosqualene to clavaric acid. Mutants disrupted in occ by gene replacement were unable to synthesize clavaric acid, but did not require sterols for growth. Amplification of the occ gene produced a 35-67% increase in clavaric acid yield. Northern analysis of occ gene expression confirmed that there is a clear correlation of occ expression and clavaric acid biosynthesis. Analysis of the occ-encoded oxidosqualene cyclase revealed that it has VSDCVGE motif instead of the consensus VSDCTAE sequence of the active center. In summary, there is an oxidosqualene cyclase specific for secondary metabolite biosynthesis; this is in agreement with the finding of two squalene cyclases in the sequenced genomes of basidiomycetes. PMID- 19130893 TI - Decreased expression of calpain and calpastatin mRNA during development is highly correlated with muscle protein accumulation in neonatal pigs. AB - It is well known that rapid gain of muscle mass in neonatal pigs is highly related to protein synthesis. However, the role of protein degradation in muscle gain of the neonatal period has not been well established. Calpains and their endogenous inhibitors, calpastatins, play a significant role in early-stage myofibrillar protein degradation. To investigate the role of calpain-calpastatin system in muscle protein accumulation, we studied the expressions of their mRNA in muscle tissue sampled at days 1, 4, 6, 12, 20 and 28 from a total of 36 neonatal pigs. The steady-state mRNA levels of calpains 1A, 2 and 3A, calpastatin types 1, 2 and 3, obtained by quantitative real-time PCR analysis, decreased by 2 4 folds at the age of 4 to 6 days compared to 1-day-old piglets. Then, the relatively low expression level was maintained through 28 days of age. Expressions of calpains 1A, 3A and calpastatin type 1 were significantly correlated with the measurements of muscle protein accumulations such as muscle protein content and RNA/protein ratio. Expressions of calpain 1A, calpastatin types 1 and 3 were negatively correlated with birth weight and fractional rate of growth. The levels of calpains 1A and 2 mRNA were correspondent to their protease activities. In conclusion, decreased levels of calpain and calpastatin expressions over development in neonatal pigs are associated with high protein accumulations, suggesting that dramatic muscle growth during the neonatal period may be partially controlled by down-regulated calpain-calpastatin system. PMID- 19130894 TI - On the biosynthesis and biological role of eoxins and 15-lipoxygenase-1 in airway inflammation and Hodgkin lymphoma. AB - This mini-review is focused on the enzyme 15-lipoxygenase-1 (15-LO-1) and eoxins in airway inflammatory diseases and Hodgkin lymphoma. Several studies have demonstrated increased expression and activity of 15-LO-1 in the respiratory tissue from asthma patients , indicating a pathophysiological role of this enzyme in airway inflammation. Eoxins were recently identified as pro-inflammatory metabolites of arachidonic acid, formed through the 15-LO-1 pathway, in human eosinophils, mast cells, airway epithelial cells and Hodgkin lymphoma. Mice deficient of 12/15-LO, the ortholog to human 15-LO-1, had an attenuated allergic airway inflammation compared to wild type controls, also indicating a pathophysiological role of this enzyme in respiratory inflammation. The putative therapeutic implications of 15-LO-1 inhibitors in the treatment of asthma, chronic obstructive pulmonary disorder and Hodgkin lymphoma are discussed. PMID- 19130895 TI - Enzymatically inactive adenylate kinase 4 interacts with mitochondrial ADP/ATP translocase. AB - Adenylate kinase 4 (AK4) is a unique member with no enzymatic activity in vitro in the adenylate kinase (AK) family although it shares high sequence homology with other AKs. It remains unclear what physiological function AK4 might play or why it is enzymatically inactive. In this study, we showed increased AK4 protein levels in cultured cells exposed to hypoxia and in an animal model of the neurodegenerative disease amyotrophic lateral sclerosis. We also showed that short hairpin RNA (shRNA)-mediated knockdown of AK4 in HEK293 cells with high levels of endogenous AK4 resulted in reduced cell proliferation and increased cell death. Furthermore, we found that AK4 over-expression in the neuronal cell line SH-SY5Y with low endogenous levels of AK4 protected cells from H(2)O(2) induced cell death. Proteomic studies revealed that the mitochondrial ADP/ATP translocases (ANTs) interacted with AK4 and higher amount of ANT was co precipitated with AK4 when cells were exposed to H(2)O(2) treatment. In addition, structural analysis revealed that, while AK4 retains the capability of binding nucleotides, AK4 has a glutamine residue instead of a key arginine residue in the active site well conserved in other AKs. Mutation of the glutamine residue to arginine (Q159R) restored the adenylate kinase activity with GTP as substrate. Collectively, these results indicate that the enzymatically inactive AK4 is a stress responsive protein critical to cell survival and proliferation. It is likely that the interaction with the mitochondrial inner membrane protein ANT is important for AK4 to exert the protective benefits to cells under stress. PMID- 19130896 TI - Human apolipoprotein A-I binds amyloid-beta and prevents Abeta-induced neurotoxicity. AB - Aggregates of the amyloid-beta peptide (Abeta) play a central role in the pathogenesis of Alzheimer's disease (AD). Identification of proteins that physiologically bind Abeta and modulate its aggregation and neurotoxicity could lead to the development of novel disease-modifying approaches in AD. By screening a phage display peptide library for high affinity ligands of aggregated Abeta(1 42), we isolated a peptide homologous to a highly conserved amino acid sequence present in the N-terminus of apolipoprotein A-I (apoA-I). We show that purified human apoA-I and Abeta form non-covalent complexes and that interaction with apoA I affects the morphology of amyloid aggregates formed by Abeta. Significantly, Abeta/apoA-I complexes were also detected in cerebrospinal fluid from AD patients. Interestingly, apoA-I and apoA-I-containing reconstituted high density lipoprotein particles protect hippocampal neuronal cultures from Abeta-induced oxidative stress and neurodegeneration. These results suggest that human apoA-I modulates Abeta aggregation and Abeta-induced neuronal damage and that the Abeta binding domain in apoA-I may constitute a novel framework for the design of inhibitors of Abeta toxicity. PMID- 19130897 TI - Transcriptional upregulation of p19INK4d upon diverse genotoxic stress is critical for optimal DNA damage response. AB - p19INK4d promotes survival of several cell lines after UV irradiation due to enhanced DNA repair, independently of CDK4 inhibition. To further understand the action of p19INK4d in the cellular response to DNA damage, we aimed to elucidate whether this novel regulator plays a role only in mechanisms triggered by UV or participates in diverse mechanisms initiated by different genotoxics. We found that p19INK4d is induced in cells injured with cisplatin or beta-amyloid peptide as robustly as with UV. The mentioned genotoxics transcriptionally activate p19INK4d expression as demonstrated by run-on assay without influencing its mRNA stability and with partial requirement of protein synthesis. It is not currently known whether DNA damage-inducible genes are turned on by the DNA damage itself or by the consequences of that damage. Experiments carried out in cells transfected with distinct damaged DNA structures revealed that the damage itself is not responsible for the observed up-regulation. It is also not known whether the increased expression of DNA-damage-inducible genes is related to immediate protective responses such as DNA repair or to more delayed responses such as cell cycle arrest or apoptosis. We found that ectopic expression of p19INK4d improves DNA repair ability and protects neuroblastoma cells from apoptosis caused by cisplatin or beta-amyloid peptide. Using clonal cell lines where p19INK4d levels can be modified at will, we show that p19INK4d expression correlates with increased survival and clonogenicity. The results presented here, prompted us to suggest that p19INK4d displays an important role in an early stage of cellular DNA damage response. PMID- 19130898 TI - Expression of phospholipases A2 in primary human lung macrophages: role of cytosolic phospholipase A2-alpha in arachidonic acid release and platelet activating factor synthesis. AB - Macrophages are a major source of lipid mediators in the human lung. Expression and contribution of cytosolic (cPLA(2)) and secreted phospholipases A(2) (sPLA(2)) to the generation of lipid mediators in human macrophages are unclear. We investigated the expression and role of different PLA(2)s in the production of lipid mediators in primary human lung macrophages. Macrophages express the alpha, but not the zeta isoform of group IV and group VIA cPLA(2) (iPLA(2)). Two structurally-divergent inhibitors of group IV cPLA(2) completely block arachidonic acid release by macrophages in response to non-physiological (Ca(2+) ionophores and phorbol esters) and physiological agonists (lipopolysaccharide and Mycobacterium protein derivative). These inhibitors also reduce by 70% the synthesis of platelet-activating factor by activated macrophages. Among the full set of human sPLA(2)s, macrophages express group IIA, IID, IIE, IIF, V, X and XIIA, but not group IB and III enzymes. Me-Indoxam, a potent and cell impermeable inhibitor of several sPLA(2)s, has no effect on arachidonate release or platelet activating factor production. Agonist-induced exocytosis is not influenced by cPLA(2) inhibitors at concentrations that block arachidonic acid release. Our results indicate that human macrophages express cPLA(2)-alpha, iPLA(2) and several sPLA(2)s. Cytosolic PLA(2)-alpha is the major enzyme responsible for lipid mediator production in human macrophages. PMID- 19130900 TI - A review of sudden unexpected death in epilepsy: prediction of patients at risk. AB - This review attempts to provide up-to-date quantitative data from published reports on sudden unexpected death in epilepsy (SUDEP) appearing on Medline and, especially, to provide a means to predict the probability of SUDEP in a given patient. The mean incidence of SUDEP was 1.8/1000, similar to the median of 1.5. The mean standardized mortality ratio was 6.8, and the mean percentage of SUDEP cases among deaths from epilepsy was 16.6. Seventeen risk factors were identified, each given a value according to the number of studies in the literature that specified that condition as a significant risk. The addition of these 17 values then indicated the risk for a given patient. The author calculated these for a group of 91 patients who died of SUDEP and also for 91 live patients. Many of their values for the different risks were significantly different. The sensitivity of these SUDEP values was 71.3%, the specificity 81.8%, and the positive predictive value 84.6%. A discussion includes the question of whether the death in SUDEP is primarily cardiac or pulmonary and the suggestion that it may be either or both in a given patient. The most important risk factor in this study was noncompliance with antiepileptic medication, and the main message of this study to caregivers is that therapeutic drug levels are crucial to avoid SUDEP. PMID- 19130901 TI - Intravenous lacosamide as successful treatment for nonconvulsive status epilepticus after failure of first-line therapy. AB - Treatment of status epilepticus usually requires intravenous anticonvulsant therapy. Lacosamide is a novel anticonvulsant drug that is available as infusion solution. We describe a patient with nonconvulsive status epilepticus who was successfully treated with intravenous lacosamide. PMID- 19130899 TI - Subjective perception of cognition is related to mood and not performance. AB - OBJECTIVE: Clinicians monitor cognitive effects of drugs primarily by asking patients to describe their side effects. We examined the relationship of subjective perception of cognition to mood and objective cognitive performance in healthy volunteers and neurological patients. METHODS: Three separate experiments used healthy adults treated with lamotrigine (LTG) and topiramate (TPM), adults with epilepsy on LTG or TPM, and patients with idiopathic Parkinson's disease. Correlations were calculated for change scores on and off drugs in the first two experiments and for the single assessment in Experiment 3. RESULTS: Across all three experiments, significant correlations were more frequent (chi(2)=259, P < or = 0.000) for mood versus subjective cognitive perception (59%) compared with subjective versus objective cognition (2%) and mood versus objective cognitive performance (2%). CONCLUSIONS: Subjective perception of cognitive effects is related more to mood than objective performance. Clinicians should be aware of this relationship when assessing patients' cognitive complaints. PMID- 19130902 TI - Adaptive trial design: a general methodology for censored time to event data. AB - Adaptive designs allow a clinical trial design to be changed according to interim findings without inflating type I error. The Inverse Normal method can be considered as an adaptive generalization of classical group sequential designs. The use of the Inverse Normal method for censored survival data was demonstrated only for the logrank statistic. However, the logrank statistic is inefficient in the presence of nuisance covariates affecting survival. We demonstrate, how the Inverse Normal method can be applied to Cox regression analysis. The required independence between test statistics of the different stages of the trial can be obtained by two different approaches. One is using the independent increment structure of the score process. The other uses right censoring and left truncating to divide individuals follow-up into per-stage data. Simulation studies show, that performance of the adaptive design does not depend on the method used for obtaining independence. Either way, an adaptive Cox regression analyis is more efficient than an adaptive logrank analysis if nuisance covariates affect survival. PMID- 19130904 TI - Simvastatin attenuates leukocyte recruitment in experimental bacterial meningitis. AB - Statins exert multiple effects besides lowering the serum cholesterol level, which might be beneficial for patients with sepsis and infections. We designed this study to assess the therapeutic potential of simvastatin in an established animal model of pneumococcal meningitis, a disease characterized by high morbidity and mortality despite effective antibiotic treatment. 24 h after injection of live pneumococci into the cisterna magna of mice, animals were clinically evaluated, cerebrospinal fluid (CSF) leukocyte counts and intracranial pressure were determined, and brains were removed for assessment of bacterial titer and blood-brain barrier breakdown. The following experimental groups were investigated: (I) no infection and treatment with vehicle; (II) infection and treatment with vehicle; infection and treatment with (III) 20 mg/kg or (IV) 40 mg/kg simvastatin s.c. 18 h before and just prior to infection. Treatment with simvastatin dose-dependently decreased CSF leukocyte counts, a marker for CNS inflammation. In addition, hypothermia was completely abolished in the 40 mg/kg simvastatin group. In contrast, a neurological clinical score, and intracranial complications like increase in intracranial pressure and blood-brain barrier breakdown were not altered by the treatment. In conclusion, simvastatin attenuates CNS leukocyte recruitment and systemic complications of experimental pneumococcal meningitis. PMID- 19130903 TI - Operations and pelvic muscle training in the management of apical support loss (OPTIMAL) trial: design and methods. AB - The primary aims of this trial are: 1) to compare surgical outcomes following sacrospinous ligament fixation to uterosacral vaginal vault suspension in women undergoing vaginal surgery for apical or uterine pelvic organ prolapse and stress urinary incontinence and 2) to examine the effects of a structured perioperative program consisting of behavioral techniques and pelvic floor muscle training compared to usual care. This trial is performed through the Pelvic Floor Disorders Network (PFDN), which is funded by National Institute of Child Health and Human Development. Subjects will be enrolled from hospitals associated with seven PFDN clinical centers across the United States. A centralized biostatistical coordinating center will oversee data collection and analysis. Two approaches will be investigated simultaneously using a 2x2 randomized factorial design: a surgical intervention (sacrospinous ligament fixation versus uterosacral vaginal vault suspension) and a perioperative behavioral intervention (behavioral and pelvic floor muscle training versus usual care). Surgeons have standardized essential components of each surgical procedure and have met specific standards of expertise. Providers of the behavioral intervention have undergone standardized training. Anatomic, functional, and health-related quality of life outcomes will be assessed using validated measures by researchers blinded to all randomization assignments. Cost-effectiveness analysis will be performed using prospectively collected data on health care costs and resource utilization. The primary surgical endpoint is a composite outcome defined by anatomic recurrence, recurrence of bothersome vaginal prolapse symptoms and/or retreatment and will be assessed 2 years after the index surgery. Endpoints for the behavioral intervention include both short-term (6-month) improvement in urinary symptoms and long-term (2-year) improvement in anatomic outcomes and prolapse symptoms. This article describes the rationale and design of this randomized trial, focusing on several key design features of potential interest to researchers in the field of female pelvic floor disorders and others conducting randomized surgical trials. PMID- 19130905 TI - Structures of AcrR and CmeR: insight into the mechanisms of transcriptional repression and multi-drug recognition in the TetR family of regulators. AB - The transcriptional regulators of the TetR family act as chemical sensors to monitor the cellular environment in many bacterial species. To perform this function, members of the TetR family harbor a diverse ligand-binding domain capable of recognizing the same series of compounds as the transporters they regulate. Many of the regulators can be induced by a wide array of structurally unrelated compounds. Binding of these structurally unrelated ligands to the regulator results in a conformational change that is transmitted to the DNA binding region, causing the repressor to lose its DNA-binding capacity and allowing for the initiation of transcription. The multi-drug binding proteins AcrR of Escherichia coli and CmeR from Campylobacter jejuni are members of the TetR family of transcriptional repressors that regulate the expression of the multidrug resistant efflux pumps AcrAB and CmeABC, respectively. To gain insights into the mechanisms of transcriptional regulation and how multiple ligands induce the same physiological response, we determined the crystal structures of the AcrR and CmeR regulatory proteins. In this review, we will summarize the new findings with AcrR and CmeR, and discuss the novel features of these two proteins in comparison with other regulators in the TetR family. PMID- 19130907 TI - Gly192 at hinge 2 site in the chaperonin GroEL plays a pivotal role in the dynamic apical domain movement that leads to GroES binding and efficient encapsulation of substrate proteins. AB - The subunit structure of chaperonin GroEL is divided into three domains; the apical domain, the intermediate domain, and the equatorial domain. Each domain has a specific role in the chaperonin mechanism. The 'hinge 2' site of GroEL contains three glycine residues, Gly192, Gly374, and Gly375, connecting the apical domain and the intermediate domain. In this study, to understand the importance of the hinge 2 amino acid residues in chaperonin function, we substituted each of these three glycine residues to tryptophan. The GroEL mutants G374W and G375W were functionally similar to wild-type GroEL. However, GroEL G192W showed a significant decrease in the ability to assist the refolding of stringent substrate proteins. Interestingly, from biochemical assays and characterization using surface plasmon resonance analysis, we found that GroEL G192W was capable of binding GroES even in the absence of ATP to form a very stable GroEL-GroES complex, which could not be dissociated even upon addition of ATP. Electron micrographs showed that GroEL G192W intrinsically formed an asymmetric double ring structure with one ring locked in the 'open' conformation, and it is postulated that GroES binds to this open ring in the absence of ATP. Trans-binding of both substrate protein and GroES was observed for this binary complex, but simultaneous binding of both substrate and GroES (a mechanism that ensures substrate encapsulation) was impaired. We postulate that alteration of Gly192 severely compromises an essential movement that allows efficient encapsulation of unfolded protein intermediates. PMID- 19130906 TI - Interactions of the M. tuberculosis UsfX with the cognate sigma factor SigF and the anti-anti sigma factor RsfA. AB - M. tuberculosis employs an exquisite cascade consisting of the cognate anti-sigma factor UsfX and anti-anti sigma factors RsfA and RsfB to regulate the functions of the alternate sigma factor SigF. We have purified these proteins to characterize their molecular properties and interactions with UsfX. UsfX forms a stable complex with SigF that could be purified only after co-expressing the proteins in E.coli. Formation of the complex is nucleotide independent and apparently requires unknown in vivo factors. Fluorescence spectroscopy experiments suggest that the nucleotide binding sites of UsfX are distal to the protein-protein interaction interface. RsfA is a novel anti-anti sigma factor whose binding to UsfX is triggered by the reduction of an intrachain disulphide bond between Cys73-Cys109. The reduction is accompanied by an increase in the hydrodynamic radius of the protein. The UsfX-RsfA complex exhibits a novel stoichiometry of 2:1 compared to the 2:2 stoichiometry reported for other anti anti-sigma factors. The role of the disulphide bond in complex formation was explored using molecular dynamics simulations. These studies support specific conformational changes that occur upon reduction of the Cys73-Cys109 bond of RsfA. This leads to a rearrangement that increases the interactions of a conserved His107 of UsfX with Cys109 of RsfA. PMID- 19130909 TI - Efficacy and safety of pharmacological agents in managing osteoporosis in the old old: review of the evidence. AB - INTRODUCTION: Osteoporosis and fracture risk increase exponentially in postmenopausal females. This places a significant burden in terms of morbidity, mortality and costs that are likely to increase with an ageing population. Despite this there is very limited data on pharmacological management of osteoporosis in this high risk group. OBJECTIVES OF THIS REVIEW: To review the published literature on the clinical efficacy and safety of specific anti osteoporosis treatments in the reduction in fracture risk in females >or=75 years of age. The following major endpoints were used in this review: SEARCH METHODS FOR IDENTIFICATION OF STUDIES: We performed an electronic search of Medline (1970 to June 2007) and the Cochrane Library (1996 to June 2007). Our search strategy included MeSH terms for osteoporosis and treatments. We reviewed the reference list of identified articles for additional relevant published trials. RESULTS: Two hundred and fifty-two potentially relevant abstracts were identified. Only six publications were deemed to meet full eligibility criteria and one met most criteria. There is evidence for significant vertebral fracture relative risk reduction(RR) at 1 year for Risedronate (RR 81%; p<0.001), Teriparatide (RR 65%; p<0.05) and Strontium Ranelate (RR 59%; p=0.002) and 3 years for Risedronate (RR 44%; p=0.003), Alendronate (RR 38%; p<0.05) and Strontium Ranelate (RR 32%; p=0.013). There is evidence for significant non-vertebral fracture relative risk reduction at 1 year for Strontium Ranelate (RR 41%; p=0.027) but not Teriparatide (p=0.66) and 3 years for Strontium Ranelate (RR 31%; p=0.011) but not Risedronate (p=0.66). The only study to report a reduction in hip fracture at 3 years is the TROPOS study with Strontium Ranelate (RR 36%; p=0.046). DISCUSSION: This review reinforces the irony that the least evidence is available for fragility fracture reduction in the group at greatest risk; the old old and those with non vertebral and hip fracture. Although there is good evidence for the benefit of the bisphosphonates (Alendronate and Risedronate), Teriparatide and Strontium Ranelate in vertebral fracture reduction, there are very limited data for non vertebral and hip fracture reduction. Strontium Ranelate is the only agent to date that has demonstrated a reduction in non vertebral and hip fracture events in this high risk elderly female population. Perhaps we need to adopt different strategies in managing older patients with osteoporosis as their fracture risks and treatment strategies may be quite different from younger populations. PMID- 19130908 TI - Ebf1-dependent control of the osteoblast and adipocyte lineages. AB - Ebf1 is a transcription factor essential for B cell fate specification and function and important for the development of olfactory sensory neurons. We show here that Ebf1 also plays an important role in regulating osteoblast and adipocyte development in vivo. Ebf1 mRNA and protein is expressed in MSCs, in OBs at most stages of differentiation, and in adipocytes. Tibiae and femora from Ebf1(-/-) mice had a striking increase in all bone formation parameters examined including the number of OBs, osteoid volume, and bone formation rate. Serum osteocalcin, a marker of bone formation, was significantly elevated in mutant mice. The numbers of osteoclasts in bone were normal in younger (4 week-old) Ebf1(-/-) mice but increased in older (12 week-old) Ebf1(-/-) mice. This correlated well with in vitro osteoclast development from bone marrow cells. In addition to the increased osteoblastogenesis, there was a dramatic increase in adipocyte numbers in the bone marrow of Ebf1(-/-) mice. Increased adiposity was also seen histologically in the liver but not in the spleen of these mice, and accompanied by decreased deposition of adipose to subcutaneous sites. Thus Ebf1 deficient mice appear to be a new model of lipodystrophy. Ebf1 is a rare example of a transcription factor that regulates both the osteoblast and adipocyte lineages similarly. PMID- 19130911 TI - Injury among Indigenous people - charting a safer future: a symposium at the Royal Australasian College of Surgeons. PMID- 19130910 TI - Identify fracture-critical regions inside the proximal femur using statistical parametric mapping. AB - We identified regions inside the proximal femur that are most strongly associated with hip fracture. Bone densitometry based on such fracture-critical regions showed improved power in discriminating fracture patients from controls. INTRODUCTION: Hip fractures typically occur in lateral falls, with focal mechanical failure of the sub-volumes of tissue in which the applied stress exceeds the strength. In this study, we describe a new methodology to identify proximal femoral tissue elements with highest association with hip fracture. We hypothesize that bone mineral density (BMD) measured in such sub-volumes discriminates hip fracture risk better than BMD in standard anatomic regions such as the femoral neck and trochanter. MATERIALS AND METHODS: We employed inter subject registration to transform hip QCT images of 37 patients with hip fractures and 38 age-matched controls into a voxel-based statistical atlas. Within voxels, we performed t-tests between the two groups to identify the regions which differed most. We then randomly divided the 75 scans into a training set and a test set. From the training set, we derived a fracture-driven region of interest (ROI) based on association with fracture. In the test set, we measured BMD in this ROI to determine fracture discrimination efficacy using ROC analysis. Additionally, we compared the BMD distribution differences between the 29 patients with neck fractures and the 8 patients with trochanteric fractures. RESULTS: By evaluating fracture discrimination power based on ROC analysis, the fracture-driven ROI had an AUC (area under curve) of 0.92, while anatomic ROIs (including the entire proximal femur, the femoral neck, trochanter and their cortical and trabecular compartments) had AUC values between 0.78 and 0.87. We also observed that the neck fracture patients had lower BMD (p=0.014) in a small region near the femoral neck and the femoral head, and patients with trochanteric fractures had lower BMD in trochanteric regions such as in the internal calcar septum (p=0.006). CONCLUSIONS: We have identified the sub-volumes of proximal femoral tissue which have the strongest association with hip fracture. The power to predict fracture can be improved, by focusing on BMD measurements in the fracture-critical regions, rather than in standard ROIs. PMID- 19130912 TI - Indigenous trauma: a New Zealand perspective. AB - The study of indigenous health is an emerging specialty and differs from other health disciplines in that the traditions and beliefs of indigenous people must be considered in developing health policy. New Zealand Maori and Australian Aboriginal and Torres Straight Islanders face similar health issues but have followed a different historical path. New Zealand was settled by the Maori from Eastern Polynesia in approximately 1300AD. A structured and settled society had developed by the time of British colonization in the 1800s. Because of Maori's obvious sovereignty over New Zealand the British negotiated a treaty in 1840 (The Treaty of Waitangi) with Maori that gave provision for their rights as British and later New Zealand citizens. Maori health indices suffered after the start of colonization but slowly rebounded in the 1900s linked to a resurgence in Maori culture, sporting and combat achievements. A sustained period of protest in the years following World War II has resulted in the inclusion of the provisions of the Treaty of Waitangi in legislation. Historical grievances of Maori relating to land confiscations and injustice are being addressed with formal apology and compensation. This process has allowed Maori to create their own infrastructure, to begin to develop their own health-care initiatives and to advise health-care authorities and governments on interventions to reduce health disparities between Maori and non-Maori New Zealanders. PMID- 19130913 TI - Understanding the extent and impact of Indigenous road trauma. AB - While transport-related injuries represent a leading cause of death for all Australians, several disparities exist between Indigenous and non-Indigenous populations. Compared to non-Indigenous Australians, Indigenous Australians are more than twice as likely to have a transport-related fatal injury, with fatality risk remaining high with age while declining among the non-Indigenous community. Few studies and appropriate databases exist to elucidate the causes and solutions to this over-representation. This paper presents a summary of known risk factors, discusses the impact of road injury at multiple levels and suggests steps towards addressing this significant public health problem. PMID- 19130914 TI - Indigenous family violence: a statistical challenge. AB - The issue of family violence and sexual abuse in Indigenous communities across Australia has attracted much attention throughout 2007, including significant intervention by the federal government into communities deemed to be in crisis. This paper critically examines the reporting and recording of Indigenous violence in Australia and reflects on what 'statistics' can offer as we grapple with how to respond appropriately to a problem defined as a 'national emergency'. PMID- 19130915 TI - The growing burden of injuries and trauma in Alice Springs. AB - In Alice Springs, assault and attempted homicide, self-harm and attempted suicide and transportation accidents contribute substantially to the burden of disease, especially among the aboriginal population who are poor and disadvantaged. While road traffic accidents and self-inflicted injuries are the leading causes of injury-related deaths worldwide, violence is a major factor in Alice Springs trauma. Violence accounted for more than half the annual trauma case load. Aboriginal Central Australians bear a disproportionate risk of injury and illness compared to their non-aboriginal counterparts. Rampant alcoholism and social and family breakdown are thought to be significant contributors to the high incidence of violence in Alice Springs. There were 2,800 trauma admissions to Alice Springs hospital in 2006 compared to 1,800 admissions in 2003. Geographical location often limits timeliness, access and level of health care available to rural and isolated regions of central Australia. Solutions to the trauma epidemic in our Remote indigenous population must look past the Emergency and Surgical Departments that care for the injured to the individual, community, environmental, social and economic factors that underpin the traumas. Traumatic injury and death maybe the most preventable of all health issues. While tremendous resources are spent caring for injured patients in hospital, less attention is paid towards gaining a better understanding of injury prevention. PMID- 19130916 TI - Injury and trauma issues from a primary health care perspective. AB - Trauma continues to be a significant health issue for Aboriginal and Torres Strait Islander peoples and communities. Data related to trauma, covering a range of issues, shows poor outcomes for Aboriginal and Torres Strait Islander patients. The context of trauma in Aboriginal and Torres Strait Islander communities and the effect of social issues must be understood in order to effectively address trauma at clinical, health system, and public health levels. PMID- 19130917 TI - The challenges of developing a trauma system for Indigenous people. AB - Trauma systems have been shown to provide the best trauma care for injured patients. A trauma system developed for Indigenous people should take into account many factors including geographical remoteness and cultural diversity. Indigenous people suffer from a significant intentional and non-intentional burden of injury, often greater than non-Indigenous populations, and a public health approach in dealing with trauma can be adopted. This includes transport issues, prevention and control of intentional violence, cultural sensitization of health providers, community emergency responses, community rehabilitation and improving resilience. The ultimate aim is to decrease the trauma burden through a trauma system with which indigenous people can fully identify. PMID- 19130918 TI - An analysis of national health strategies addressing Indigenous injury: consistencies and gaps. AB - This paper reviews the inclusion of injury in national Aboriginal and Torres Strait Islander health strategy. The review focused on strategies published during the period 2003 2008 or strategies which are otherwise identified as current for this period. Their content in relation to injury and its antecedents is described. This content is evaluated using the National Aboriginal and Torres Strait Islander Safety Promotion Strategy as a benchmark. The review demonstrates that injury is recognised as a public health priority across Indigenous health strategy documents, and strategies that address social and emotional wellbeing (including violence and one of its antecedents, alcohol and substance misuse) are clearly reinforced across national strategy in Indigenous health. However, there are some significant gaps and fragmentation of injury strategy remains a continuing challenge in this field. PMID- 19130919 TI - Injury prevention in Australian Indigenous communities. AB - Injury prevention in Indigenous communities in Australia is a continuing national challenge, with Indigenous fatality rates due to injury three times higher than the general population. Suicide and transport are the leading causes of injury mortality, and assault, transport and falls the primary causes of injury morbidity. Addressing the complex range of injury problems in disadvantaged Indigenous communities requires considerable work in building or enhancing existing capacity of communities to address local safety issues. Poor data, lack of funding and absence of targeted programs are some of the issues that impede injury prevention activities. Traditional approaches to injury prevention can be used to highlight key areas of need, however adaptations are needed in keeping with Indigenous peoples' holistic approach to health, linked to land and linked to community in order to address the complex spiritual, emotional and social determinants of Indigenous injury. PMID- 19130920 TI - The Australian indigenous health program. AB - This paper is an evaluation of an intercollegiate program that targeted specialist surgeons, obstetricians and gynaecologists treating Australian Indigenous patients. The program was a continuing professional development activity that was delivered through on-line modules and focussed on providing the specialist with practical skills to improve the health of Indigenous Australians in a clinical setting. PMID- 19130921 TI - The building blocks of a professional response to Indigenous injury. PMID- 19130922 TI - Gene-environment interactions in asthma. AB - Asthma is a complex disease, and its incidence is determined by an intricate interplay of genetic and environmental factors. The identification of novel genes for asthma suggests that many genes with small effects rather than few genes with strong effects contribute to the development of asthma. These genetic effects may in part differ with respect to a subject's environmental exposures, although some genes may also exert their effect independently of the environment. Whereas the geneticist uses highly advanced, rapid, comprehensive technologies to assess even subtle changes in the human genome, the researcher interested in environmental exposures is often confronted with crude information obtained from questionnaires or interviews. There is thus substantial need to develop better tools for individual exposure assessment in all relevant environmental fields. Despite these limitations, a number of important gene-environment interactions have been identified. These interactions point to the biology of environmental exposures as the involved genetic variation is suggestive of certain underlying mechanisms. Furthermore, the identification of subjects who are particularly susceptible to environmental hazards through genetic analyses helps to estimate better the strength of effect of environmental exposures. Finally, the analysis of gene environment interactions may result in a reconciliation of seemingly contradictory findings from studies not taking environmental exposures into account. PMID- 19130923 TI - Gene-environment interactions: the road less traveled by in asthma genetics. PMID- 19130924 TI - Advances in pediatric asthma in 2008: where do we go now? AB - This year's summary focuses on recent advances in pediatric asthma as reported in Journal publications in 2008. New National Asthma Education and Prevention Program asthma guidelines were released in 2007 with a special emphasis on asthma control. Attention was redirected to methods that could reduce impairment, specifically symptom control, and minimize risk, including exacerbations. Journal theme issues in 2008 focused on several relevant asthma topics including asthma exacerbations, exercise-induced bronchospasm, asthma and obesity, and occupational asthma. This review highlights Journal articles and related articles that reinforce principles of the guidelines and also direct us to new information that will advance asthma care for children. A major step forward will be finding ways to implement the asthma guidelines. PMID- 19130927 TI - Association of allergen-specific regulatory T cells with the onset of clinical tolerance to milk protein. AB - BACKGROUND: About 70% of children with milk allergy tolerate extensively heated milk (HM) products and outgrow their allergy earlier than those who react to HM. OBJECTIVE: To test the hypothesis that HM-tolerant children have a higher precursor frequency of adaptive allergen-specific regulatory T (Treg) cells. METHODS: Allergic, HM-tolerant, outgrown, or control subjects were defined by oral food challenge. PBMCs were cultured with purified caseins and controls for 7 days, and proliferating CD25(+)CD27(+) Treg cells were identified by flow cytometry. Proliferating cells were also characterized for their expression of FoxP3, CTLA 4, CD45RO, and CD127. Allergen-specific Treg cell origin and function were assessed by depletion of CD25(hi) cells before culture. RESULTS: There was a higher percentage (median [25th% to 75th%], 16.85% [7.1-31.7]) of proliferating allergen-specific CD25(+)CD27(+) T cells from cultures of HM-tolerant subjects (n = 18) than subjects with allergy (n = 8; 4.91% [2.6-7.5]; P < .01). Control subjects with no history of milk allergy (n = 7) also had low percentages of these cells (2.9% [2.4-6.0]), whereas outgrown subjects (n = 7) had intermediate percentages (9.0% [2.7-16.4]). There were no significant differences between the patient groups in the frequency of polyclonal Treg cells or allergen-specific effector T cells. Allergen-specific Treg cells were found to be FoxP3(+)CD25(hi)CD27(+), cytotoxic T lymphocyte-associated antigen 4(+), CD45RO(+)CD127(-) and were derived from circulating CD25(hi) T cells. Depletion of the CD25(hi) cells before in vitro culture significantly enhanced allergen specific effector T-cell expansion. CONCLUSION: A higher frequency of milk allergen-specific Treg cells correlates with a phenotype of mild clinical disease and favorable prognosis. PMID- 19130925 TI - Advances in adult asthma diagnosis and treatment and health outcomes, education, delivery, and quality in 2008. AB - In 2008 the Journal reported new findings in management of asthma. Dosing schedules of inhaled steroids have been modified and individualized. New, more costly propellants are replacing ozone-depleting chlorofluorocarbons. An association of asthma with pneumococcal disease has been observed. Smoking bans in public places are eliminating second-hand smoke and reducing asthma-related emergency department visits among adults. In contrast with these advances, however, disparity in asthma morbidity persists: black persons compared with white persons have a 4-fold greater risk of an asthma-related emergency department visit. PMID- 19130929 TI - The effect of obesity on asthma incidence: moving past the epidemiologic evidence. PMID- 19130928 TI - Notch signaling confers antigen-presenting cell functions on mast cells. AB - BACKGROUND: Notch signaling is involved in cell fate determination along with the development of the immune system. However, very little is known about the role for Notch signaling in mast cells. OBJECTIVE: We investigated the role of Notch signaling in mast cell functions. METHODS: After mouse bone marrow-derived mast cells (BMMCs) or peritoneal mast cells (PMCs) were cocultured with mouse Notch ligand-expressing chinese hamster ovary cells for 5 days, we examined the mast cell surface expressions of MHC-II molecules and OX40 ligand (OX40L), Fc epsilon RI-mediated cytokine production, and the effects of the mast cells on proliferation and differentiation of naive CD4(+) T cells in vitro. RESULTS: We showed that BMMCs and PMCs constitutively expressed Notch1 and Notch2 proteins on the cell surface. We also found that Delta-like 1 (Dll1)/Notch signaling induced the expression of MHC-II and upregulated the expression level of OX40L on the surface of the mast cells. Dll1/Notch signaling augmented Fc epsilon RI-mediated IL-4, IL-6, IL-13, and TNF production by BMMCs. Dll1-stimulated MHC II(+)OX40L(high) BMMCs promoted proliferation of naive CD4(+) T cells and their differentiation into T(H)2 cells producing IL-4, IL-5, IL-10, and IL-13. CONCLUSION: Dll1/Notch signaling confers the functions as an antigen-presenting cell on mast cells, which preferentially induce the differentiation of T(H)2. PMID- 19130930 TI - Learning from molecular sleuths. PMID- 19130931 TI - Asthma symptom re-emergence after omalizumab withdrawal correlates well with increasing IgE and decreasing pharmacokinetic concentrations. AB - BACKGROUND: Physicians have questioned whether omalizumab can be discontinued or the dose reduced after clinical improvement is seen in patients with severe asthma. OBJECTIVES: To examine the relationships among omalizumab, free IgE, and clinical outcomes in a randomized, placebo-controlled trial in patients with severe persistent allergic asthma following a posology based on pretreatment total IgE and body weight. METHODS: A pharmacokinetic-pharmacodynamic binding model was used to calculate free IgE, omalizumab, and total IgE concentrations during the 28-week treatment and 16-week follow-up of the INvestigation of Omalizumab in seVere Asthma TrEatment (INNOVATE) study. These were plotted against the mean changes in the total asthma symptom score, morning peak expiratory flow, and rescue medication use for physician-defined treatment responders and nonresponders. RESULTS: The model accurately fitted omalizumab and free and total IgE, allowing reconstruction of the entire time course for each patient. Free IgE was rapidly suppressed below the 50 ng/mL (20.8 IU/mL) target, although there was a notable period before clinical measures stabilized. After treatment cessation, free IgE and omalizumab returned toward baseline and, after a delay, asthma symptoms re-emerged. Model-derived omalizumab and free IgE concentrations correlated well with changes in clinical outcomes, particularly in omalizumab-treated responders. Asthma symptoms exhibited different correlations during response onset compared with response offset (hysteresis), indicative of physiological time delays between changes in IgE levels and pulmonary function. CONCLUSION: Omalizumab and free IgE correlated well with clinical symptoms. Reducing omalizumab doses below those in the dosing table cannot be recommended; the resulting increase in free IgE would cause a deterioration in asthma control. PMID- 19130932 TI - Therapeutic efficacy of omalizumab. PMID- 19130933 TI - Cardiovascular disease risk associated with asthma and respiratory morbidity might be mediated by short-acting beta2-agonists. AB - BACKGROUND: Studies examining the asthma-related risks of cardiovascular disease (CVD) events have generally used selected samples or did not control for the effects of beta(2)-agonist use, itself associated with CVD events. OBJECTIVES: We assessed the relationship between incident CVD/stroke and asthma and the effect of atopy while controlling for beta(2)-agonist use in a representative adult population cohort free of CVD at baseline. METHODS: The North West Adelaide Health Study (stage 1, n = 3812; stage 2, n = 3113) assessed spirometry, anthropometry, atopy, blood pressure, and lipid levels. Questionnaires assessed doctor-diagnosed asthma and CVD (myocardial infarction and angina)/stroke, smoking status, and demographics. Asthma was defined by self-report or FEV(1) reversibility. Current short- and long-acting beta(2)-agonist use was identified at follow-up. RESULTS: Results are expressed as odds ratios (ORs) and 95% CIs. By using multivariable logistic regression, after adjustment for risk factors, in female subjects incident CVD/stroke events were associated with asthma (OR, 3.24; 95% CI, 1.55-6.78), with no effect modification by atopy (P for interaction = .61), and with as-required short-acting beta(2)-agonist use (OR, 2.66; 95% CI, 1.06-6.61). In male subjects events were associated with daily cough/sputum (OR, 1.92; 95% CI, 1.05-3.50) and FEV(1) of less than 80% of predicted value but an FEV(1)/forced vital capacity ratio of greater than 0.70 (OR, 2.15; 95% CI, 0.91 5.09; P = .08). Although few CVD/stroke events occurred in male subjects with asthma, a significant interaction with atopic status was found (P = .05). CONCLUSIONS: Studies are required to elucidate how asthma exposes older women to excess macrovascular risk and prospectively determine the short-acting beta(2) agonist-related risk in persons without existing CVD. CVD risk in relation to atopic status of asthma also requires further investigation. PMID- 19130934 TI - The causal direction in the association between respiratory syncytial virus hospitalization and asthma. AB - BACKGROUND: Earlier studies have reported an increased risk of asthma after respiratory syncytial virus (RSV) hospitalization. Other studies found that asthmatic disposition and propensity to wheeze increase the risk of RSV hospitalization. OBJECTIVE: The current study examined the causal direction of the associations between RSV hospitalization and asthma in a population-based cohort of twins. METHODS: We conducted a prospective cohort study examining the associations between RSV hospitalization and asthma by using registry information on RSV hospitalization and asthma among 18,614 Danish twins born 1994 to 2003. The associations between RSV and asthma were examined in both directions: we examined the risk of asthma after RSV hospitalization, and the risk of RSV hospitalization in children with asthma in the same population-based cohort. RESULTS: Asthma hospitalization after RSV hospitalization was increased as much as 6-fold to 8-fold during the first 2 months after RSV hospitalization but was no longer increased 1 year later. Asthma increased the risk of RSV hospitalization by 3-fold, and the risk was not time-dependent. Analyzing these associations on the basis of asthma defined from use of inhaled corticosteroid did not materially change the risk estimates. CONCLUSION: There is a bidirectional association between severe RSV infection and asthma. Severe RSV infection is associated with a short-term increase in the risk of subsequent asthma, suggesting that RSV induce bronchial hyperresponsiveness; and asthma is associated with a long-term increased susceptibility for severe RSV disease, suggesting a host factor being responsible for the severe response to RSV infection. This suggests that severe RSV infection and asthma may share a common genetic predisposition and/or environmental exposure. PMID- 19130935 TI - Airway glutathione homeostasis is altered in children with severe asthma: evidence for oxidant stress. AB - BACKGROUND: Severe asthma is characterized by persistent airway inflammation and increased formation of reactive oxygen species. OBJECTIVES: Glutathione (GSH) is an important antioxidant in the epithelial lining fluid (ELF). We hypothesized that airway GSH homeostasis was altered in children with severe asthma and was characterized by decreased GSH and increased glutathione disulfide (GSSG) concentrations. METHODS: Bronchoalveolar lavage was obtained from 65 children with severe asthma, including 35 children with baseline airway obstruction evidenced by FEV(1) <80%. Control data were obtained from 6 children with psychogenic (habit) cough or vocal cord dysfunction undergoing diagnostic bronchoscopy and 35 healthy adult controls. GSH, GSSG, and other determinants of airway oxidative stress including glutathione S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GPx), malondialdehyde, 8-isoprostane, and H(2)O(2) were measured in the ELF. The ELF redox potential was calculated from GSH and GSSG by using the Nernst equation. RESULTS: Compared with controls, subjects with severe asthma had lower airway GSH with increased GSSG despite no differences in GST, GR, and GPx activities between groups. This was accompanied by increased malondialdehyde, 8-isoprostane, and H(2)O(2) concentrations in the ELF. GSH oxidation was most apparent in subjects with severe asthma with airway obstruction and was supported by an upward shift in the ELF GSH redox potential. CONCLUSION: Children with severe asthma have increased biomarkers of oxidant stress in the ELF that are associated with increased formation of GSSG and a shift in the GSH redox potential toward the more oxidized state. PMID- 19130937 TI - Safety and efficacy in children of an SQ-standardized grass allergen tablet for sublingual immunotherapy. AB - BACKGROUND: Immunotherapy with the SQ-standardized grass tablet Grazax is efficacious and well-tolerated in adult patients with rhinoconjunctivitis. Allergic asthma and rhinoconjunctivitis are closely linked, and a strategy combining treatment of the upper and lower airways is recommended. OBJECTIVE: To investigate the efficacy of treatment with the grass tablet on grass pollen induced rhinoconjunctivitis and asthma as well as the immunologic response and the safety profile in children. METHODS: A total of 253 children age 5 to 16 years, with grass pollen-induced rhinoconjunctivitis with/without asthma, were randomized 1:1 to active treatment or placebo. Treatment was initiated 8 to 23 weeks before the start of the grass pollen season 2007 and continued throughout the entire season. Symptomatic medication was provided as relief medication to both groups in a stepwise fashion. Primary endpoints were rhinoconjunctivitis symptom and medication scores. RESULTS: The rhinoconjunctivitis symptom and medication scores and the asthma symptom score were all statistically significantly different between the 2 treatment groups. The differences in medians relative to placebo were 24%, 34%, and 64% in favor of active treatment. The immunologic response was similar to that observed in adults. The most common adverse reaction was oral pruritus, reported by 40 subjects (32%) in the active and 3 (2%) in the placebo group. Six subjects withdrew because of adverse events. No serious adverse events were assessed as treatment-related. CONCLUSION: Immunotherapy with the grass tablet reduced grass pollen-induced rhinoconjunctivitis and asthma symptoms in a pediatric population and introduced an immunomodulatory response, consistent with treatment of the underlying allergic disease. The treatment was well tolerated. PMID- 19130936 TI - Improving asthma self-efficacy: developing and testing a pilot community-based asthma intervention for African American adults. AB - BACKGROUND: Low-income African American adults in Chicago have disproportionately high asthma morbidity and mortality rates. Interventions that improve asthma self efficacy for appropriate self-management behaviors might ultimately improve asthma control in this population. OBJECTIVE: We sought to pilot test an intervention to improve asthma self-efficacy for appropriate self-management behaviors. METHODS: Participants for this trial were recruited through 2 primary care clinics located in the largest African American community in Chicago. Participants were then randomized into one of 2 groups. The control group received mailed asthma education materials. The intervention group was offered 4 group sessions led by a community social worker and 6 home visits by community health workers. Telephone interviews were conducted at baseline (before intervention), 3 months (after intervention), and 6 months (maintenance). RESULTS: The 42 participants were predominantly African American and low income and had poorly controlled persistent asthma. The intervention group had significantly higher asthma self-efficacy at 3 months (P < .001) after the completion of the intervention. Asthma action plans were more common in the intervention group at 3 months (P = .06). At 6 months, the intervention group had improved asthma quality of life (P = .002) and improved coping (P = .01) compared with control subjects. Trends in behavioral and clinical outcomes favored the intervention group but were not statistically significant. CONCLUSIONS: This community-based asthma intervention improved asthma self-efficacy, self-perceived coping skills, and asthma quality of life for low-income African American adults. Larger trials are needed to test the efficacy of this intervention to reduce asthma morbidity in similar high-risk populations. PMID- 19130939 TI - Angioedema and estrogen-dependent angioedema with activation of the contact system. PMID- 19130938 TI - Mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 dependent pathways are essential for CD8+ T cell-mediated airway hyperresponsiveness and inflammation. AB - BACKGROUND: Ligation of the leukotriene B(4) (LTB(4)) receptor 1 on effector memory CD8(+) T cells by LTB(4) is important for the recruitment of CD8(+) T cells into the airways, which appears central to the induction of airway hyperresponsiveness (AHR) and allergic inflammation. Phosphorylation of extracellular signal-regulated kinase (ERK) is important in activation and cytokine production from many cell types. OBJECTIVE: The roles of ERKs in effector CD8(+) T-cell function and on CD8(+) T cell-mediated AHR were determined. METHODS: Effector CD8(+) T cells were generated from OVA(257-264) (SIINFEKL) peptide-primed mononuclear cells from OT-1 mice. The effects of U0126, an ERK inhibitor, on effector CD8(+) T-cell function and on CD8(+) T cell mediated AHR and allergic inflammation were examined. RESULTS: Pretreatment of effector CD8(+) T cells with U0126 suppressed anti-CD3/anti-CD28-induced ERK1/2 phosphorylation and cytokine production, but did not affect LTB(4)-induced Ca(2+) mobilization or chemotaxis. Adoptive transfer of U0126-treated CD8(+) T cells into sensitized mice before secondary allergen challenge resulted in significant decreases in AHR, eosinophilic inflammation, goblet cell metaplasia, and IL-5 and IL-13 levels in bronchoalveolar lavage fluid of recipient mice. The number of transferred CD8(+) T cells accumulating in bronchoalveolar lavage fluid or lungs was unaffected by treatment. CONCLUSION: ERK1/2-dependent pathways are essential for the effector functions of CD8(+) T cells, including T(H)2 cytokine production, allergic inflammation, and development of AHR. Inhibition of ERK1/2 signaling has potential therapeutic benefit in preventing CD8(+) T cell-mediated AHR. PMID- 19130940 TI - Asthma and obesity: an archival addendum. PMID- 19130942 TI - Climate change and plant food allergens. PMID- 19130944 TI - Sublingual immunotherapy with venom is not recommended for patients with Hymenoptera venom allergy. PMID- 19130946 TI - Levalbuterol kinetics. PMID- 19130950 TI - Synergy. PMID- 19130951 TI - Practical guide to efficient analysis and diagramming articles. AB - OBJECTIVE: In a busy practice, time is a commodity in rare supply, and keeping abreast of the relevant medical literature is a daunting task. Even after reading an article carefully, important information may be lost because of undue attention to the methodological minutia. METHOD: Fundamental to reducing a complex article into a well-organized and consistent format is the technique of quantitatively diagramming the principal components in the design and findings of the study. This approach allows a coherent brief summary statement of the article, and a platform for a focused scientific discussion and analysis of the clinical applicability of the findings. CONCLUSIONS: The technique of diagramming an article is a tool that the senior author has used for years and has found it valuable in capturing the scientific fundamentals of a manuscript. Once prepared, the diagram has proven to be useful in clarifying presentations during journal club, preparing the background and significance sections of grant applications, reviewing articles as a journal editor and as a journal reviewer, and easily retrieving documents to support evidence-based practice efforts. PMID- 19130952 TI - The Medicine and GERD of Immanuel Kant (1724-1804). AB - We can place Kant as one of the pillars of contemporary medicine. Firstly, as an Illuminist, his work subordinates the collection of empirical data, which in medical science is constitutional to reason. This was the basis of a rational medical science. Secondly, he is the father of medical regulation, having set the philosophical control ground stone for physicians by the State. His work "Critique of Practical Reason" drafts all the future codes of ethics and bioethics. We will hereby study his relationship with medicine based on the text "The Conflict with the Faculty of Medicine" and other auxiliary texts.We can find in Kant's works the description of a series of symptoms that were related to a nonspecific dyspeptic syndrome that nowadays would be diagnosed as a strong indication that he suffered from gastroesophageal reflux disease (GERD). PMID- 19130953 TI - Can plastic tracheotomy tubes harm baby boys? AB - Pediatric tracheotomy tubes are often made from polyvinyl chloride (PVC). Di(2 ethylhexyl) phthalate (DEHP) is a common additive to PVC to increase tracheotomy tube flexibility and strength. Unfortunately, DEHP is a reproductive toxicant that alters the male reproductive system in animals, in vitro tests, and limited human data. DEHP leaches from PVC tracheotomy tubes in children. The FDA and multiple international health agencies urge health care professionals to use DEHP free medical materials for vulnerable patients, especially the male infant. The observed toxicity of DEHP and the readily available alternative materials to PVC tracheotomy tubes present a compelling argument to use pediatric tracheotomy tubes made from non-PVC materials such as polyurethane or silicone. More clinical research is needed. PMID- 19130954 TI - Extracts from The Cochrane Library: Tonsillectomy for chronic/recurrent acute tonsillitis. AB - The "Cochrane Corner" is a quarterly section in the Journal that highlights systematic reviews relevant to otolaryngology-head and neck surgery, with invited commentary to highlight implications for clinical decision making. This installment features a Cochrane Review entitled "Tonsillectomy or adeno tonsillectomy versus non-surgical treatment for chronic/recurrent acute tonsillitis," which concludes modest efficacy in children for sore throat days and episodes, with greater benefit for those with more severe illness. PMID- 19130955 TI - Adult tonsillectomy: current indications and outcomes. AB - OBJECTIVE: Limited data exist with respect to the current prevalence of indications for tonsillectomy in adults. We sought to determine the indications for adult tonsillectomy and to identify factors associated with postoperative complications. METHODS: The medical records of 361 adult patients who underwent tonsillectomy from 2001 to 2007 were reviewed. RESULTS: Indications for surgery were chronic infection in 207 (57%) patients, upper airway obstruction secondary to tonsillar hypertrophy in 98 (27%) patients, and suspected neoplasm in 56 (16%) patients. Postoperative complications occurred in 54 (15%) cases with hemorrhage in 19 (5%) patients, followed by dehydration in 16 (4%) patients, and admission for postoperative oxygen desaturations in 9 (3%) patients. Hospitalization beyond 24 hours occurred in 18 (5%) cases, with emergency room visits for pain and dehydration in 16 (4%) patients, and readmission for pain control in 17 (5%) cases. Patients who underwent tonsillectomy for upper airway obstruction had an increased incidence of prolonged hospitalization or readmission (19% vs 6%; P = 0.01), whereas patients who underwent tonsillectomy for infection had an increased incidence of postoperative bleeding (6% vs 4%; P = 0.02). CONCLUSIONS: Chronic infection remains the most common indication for adult tonsillectomy in contrast to the pediatric population. Complication rates vary according to the indication for surgery. PMID- 19130956 TI - Diathermy power settings as a risk factor for hemorrhage after tonsillectomy. AB - OBJECTIVE: To investigate bipolar diathermy power settings as a risk factor for postoperative hemorrhage following tonsillectomy. STUDY DESIGN AND SETTING: A prospective cohort study was undertaken between July 2003 and September 2004 in National Health Service (NHS) and independent hospitals in England and Northern Ireland. Data were collected on patient characteristics, tonsillectomy technique, and postoperative hemorrhage within 28 days of surgery. RESULTS: Among the 9572 patients who had a tonsillectomy performed with bipolar diathermy dissection and hemostasis, the overall rate of hemorrhage was 4.6 percent and the risk of hemorrhage was not associated with the diathermy power setting. Among the 8465 patients who had tonsillectomy with cold steel dissection and bipolar diathermy hemostasis, the rate of hemorrhage increased from 1.8% in patients with the lowest power settings (6 to 8 watts) to 3.7% in those with settings above 18 watts (P value for trend = 0.005). CONCLUSION: In tonsillectomies using cold steel dissection and bipolar diathermy for hemostasis, the risk of postoperative hemorrhage becomes greater as diathermy power increases. PMID- 19130958 TI - Impact of voice and swallowing problems in the elderly. AB - OBJECTIVES: 1) Evaluate the prevalence and quality-of-life impact of voice and swallowing problems in the elderly; 2) determine treatment trends and barriers to treatment. STUDY DESIGN: Cross-sectional study of independent-living residents in two retirement communities. Prevalence of dysphonia and dysphagia, voice-related quality of life (VRQOL), 7-point Likert scale of dysphagia severity, Center for Epidemiologic Studies Depression (CES-D) scale, and barriers to treatment were collected. Spearman correlation and ANOVA statistics were performed. RESULTS: A total of 248 residents responded with a mean age of 82.4 years; 19.8 percent had dysphonia, 13.7 percent dysphagia, and 6 percent both. Respondents with more severe swallowing difficulty had greater impairment on the VRQOL (P = 0.04, Spearman correlation = -0.4). Respondents with both dysphonia and dysphagia had greater depression scores than those with neither symptom (mean CES-D score 15.5 vs 9.9, P = 0.009, ANOVA, P < 0.05, Bonferroni t test). Only 22.4 percent and 20.6 percent had sought treatment for dysphonia and dysphagia, respectively. Being unaware of treatment options and viewing voice and swallowing trouble as a normal part of aging were the most common reasons for not seeking treatment. CONCLUSIONS: Voice and swallowing problems are common in the elderly, but they are not realizing potential treatment benefits. PMID- 19130957 TI - The efficacy of preoperative screening and the treatment of methicillin-resistant Staphylococcus aureus in an otolaryngology surgical practice. AB - OBJECTIVE: To examine the efficacy of preoperative Staphylococcus aureus screening on postoperative methicillin-resistant S aureus (MRSA) infection rates in otolaryngology. STUDY DESIGN: Chart review. SUBJECTS AND METHODS: Postoperative MRSA infection rates in unscreened patients during a 1-year period were compared with infection rates in patients after preoperative S aureus screening was initiated. Colonized patients were treated with mupirocin and chlorhexidine preoperatively. RESULTS: Records of 420 patients were reviewed. In the 241 patients without screening, nine patients had S aureus infections, and there were two (0.8%) postoperative MRSA surgical-site infections. Of 179 patients after screening was initiated, 24 patients (13.4%) were colonized with S aureus and underwent preoperative treatment. There were no MRSA infections in the postoperative period. CONCLUSION: Early results show the potential benefit of preoperative S aureus screening in MRSA infection rate reduction. Although larger studies are needed, screening and treatment of MRSA colonized patients preoperatively may reduce infectious complications in otolaryngology. PMID- 19130959 TI - Laryngeal and vocal evaluation in untreated growth hormone deficient adults. AB - OBJECTIVE: To evaluate the consequences of lifetime, severe and untreated isolated growth hormone deficiency (IGHD) on vocal and laryngeal function. STUDY DESIGN: Cross-sectional. SUBJECTS AND METHODS: A total of 23 IGHD adult subjects and 22 controls were administered a questionnaire about vocal complaints and harmful voice habits, and underwent videolaryngostroboscopic examination, voice evaluation by perceptual-auditory analysis with GRBAS scale including grade of dysphonia, roughness, breathiness, asthenia and strain items, objective voice evaluation by maximum phonation time (MPT), and acoustic analysis. RESULTS: There was no difference in vocal complaints between IGHD subjects and controls. Vocal abuse and smoking were more frequent in IGHD subjects. IGHD subjects presented higher values for roughness, breathiness, and strain. Laryngopharyngeal reflux (LPR) signs and laryngeal constriction were more frequent in IGHD individuals. MPT was similar in the two groups. Fundamental frequency was higher in IGHD females and males. Harmonic to noise ratio was higher in IGHD in both genders and shimmer was lower in IGHD females. CONCLUSIONS: IGHD subjects have higher prevalence of signs of LPR and laryngeal constriction, with high pitch in both genders, which suggests a prominent role of IGHD on these parameters. PMID- 19130960 TI - Dosage changes in patients with long-term botulinum toxin use for laryngeal dystonia. AB - OBJECTIVE: To investigate the dosage consistency of botulinum toxin injections in patients with long-term treatment for laryngeal dystonia. STUDY DESIGN: Chart review. SUBJECTS AND METHODS: Patients with laryngeal dystonia who had received at least 20 injections to the thyroarytenoid muscle were selected. Change of botulinum toxin dose, patient-reported effective weeks, and intervals between injections were investigated; data from the first five injections were excluded to eliminate initial dose titration. RESULTS: Fifty-five patients with an average follow-up of 12.5 years were identified. Mean dose of botulinum toxin was 2.37 +/ 1.6 U at the sixth injection; there was a decrease in mean dose over time, which became statistically significant at the 13th injection with a mean dose of 2.02 +/- 1.16 U. Mean duration of good effect and treatment intervals showed no significant change over time. CONCLUSION: Botulinum toxin dose needed for a constant response in treatment of laryngeal dystonia decreases over time, without any accompanying change in effective weeks and intervals. PMID- 19130961 TI - Old and new aspects of middle turbinate histopathology. AB - OBJECTIVE: To study the histology of the normal and pathologic middle turbinate (MT). STUDY DESIGN: Nonrandomized, controlled study. SUBJECTS AND METHODS: Qualitative and quantitative assessment of soft tissue and bony elements of 14 normal and 10 chronic rhinosinusitis (CRS) MT samples (patients' mean CT score, 6.5 +/- 6.7). A comparison with 14 normal inferior turbinate (IT) samples was accomplished. Quantitative assessment was limited to the lateral mucosal layer (LML). RESULTS: The thickness of the LML was significantly greater in pathologic MT than in normal MT (P = 0.002). Except for veins that underwent significant increase (P = 0.036), other soft tissue constituents of the pathologic MT did not change significantly. Glands were more abundant in normal MT and veins in normal IT (P < 0.001, P = 0.001, respectively). Qualitative assessment showed larger veins in the normal IT compared with the normal MT. Inflammatory cell infiltrate, edema, and fibrosis were found in CRS. None of our MT bones exhibited osteomyelitic changes. CONCLUSIONS: The different proportion of glands suggests varied functional roles for both the MT and IT. The significant involvement of the MT in the inflammatory process of CRS and the relatively small size and paucity of veins with relatively little effect on nasal airflow justify partial excision of the MT. PMID- 19130962 TI - Microsatellite instability analysis of sinonasal carcinomas. AB - OBJECTIVES: Intestinal-type sinonasal adenocarcinoma (ITAC) and squamous cell carcinoma of the nasal cavity (SCCNC) are histopathologically but not etiologically similar to colorectal adenocarcinoma or to laryngeal squamous cell carcinoma, respectively. Microsatellite instability (MSI) is involved in both tumors. The aim of this study was to investigate a possible role for MSI in the pathogenesis of two types of nasal carcinoma. MATERIAL AND METHODS: DNA obtained from frozen tumor samples of 41 ITACs and 24 SCCNCs was analyzed for shifts in five mononucleotide microsatellite loci by multiplex PCR. RESULTS: The allelic patterns of one ITAC (2%) and five SCCNCs (21%) revealed an allelic shift for at least one of the five loci, indicating microsatellite instability. CONCLUSION: MSI may be involved in squamous cell carcinoma, but not in adenocarcinoma of the nasal cavities. PMID- 19130963 TI - Modified lateral neck lymphadenectomy: prospective randomized study comparing harmonic scalpel with clamp-and-tie technique. AB - OBJECTIVE: To compare the use of harmonic scalpel (HS) with clamp-and-tie technique to evaluate their comparative merits in modified lateral lymphadenectomy (LL). STUDY DESIGN: Prospective and randomized. SUBJECTS AND METHODS: Thirty-seven patients were recruited and divided into Group A (conventional; n = 20) and Group B (HS; n = 17). Thyroid volume, neck circumference, operative time, diameter of the tumor and lymph node, drainage volume, pain, and complications were compared. Operation consisted of thyroidectomy and LL. RESULTS: Groups were homogeneous for thyroid volume, diameter of thyroid nodule and lymph node, and neck circumference. Operative time was shorter in Group B than in Group A. The fluid collection in the vacuum between 24 and 48 hours and the increase of neck circumference were lower in Group B. Pain was significantly lower in Group B after 12 hours and decrease was faster in Group B. CONCLUSION: The use of HS during LL is as safe as conventional technique and may allow shorter operative time, lower lymphatic spillage, and faster decrease of pain. PMID- 19130964 TI - Perineural invasion of the major and minor nerves in laryngeal and hypopharyngeal cancer. AB - OBJECTIVE: To investigate the perineural invasion in patients with squamous cell carcinoma of the larynx and hypopharynx. STUDY DESIGN: A prospective study of patients with laryngeal and hypopharyngeal squamous cell carcinoma who underwent total or partial laryngectomy. METHODS: Patients with squamous cell carcinoma of the larynx and pyriform sinus who underwent laryngectomy between 2002 and 2006 in the ENT Clinic of Cluj-Napoca were investigated for histopathological identification of perineural invasion. RESULTS: The present study included 256 patients. Perineural invasion was present in 86 cases out of the 256. Perineural invasion was detected in the major nerves of only one case out of 219 patients who had undergone total laryngectomy. The difference between the mean disease free survival of patients with or without perineural invasion of the minor nerves was statistically significant (Mann-Whitney U test, P = 0.000102). The local recurrence rates in the cases with or without perineural invasion were significantly different (log-rank test, P = 0.00001). CONCLUSION: Major nerves do not constitute a way of spreading in the squamous cell carcinoma of the larynx and hypopharynx, whereas minor nerves remain a potential one. PMID- 19130965 TI - A comparison between endoscope-assisted partial parotidectomy and conventional partial parotidectomy. AB - OBJECTIVE: To report the feasibility of endoscope-assisted parotidectomy with an ultrasonic scalpel. STUDY DESIGN: Randomized clinical trial. SUBJECTS AND METHODS: A total of 38 patients were randomly assigned to conventional (20 patients) or endoscope-assisted (18 patients) partial parotidectomy. Via retrograde approach, endoscopic and surgical instruments were inserted through two 20- to 25-mm incisions (retromandibular and postauricular). RESULTS: The endoscopic surgery lasted for 108.61 +/- 11.86 minutes and the bleeding volume was significantly lower than in the conventional group. All 18 patients were satisfied with their cosmetic results; 14 patients (65%) were satisfied with the great auricular nerve preservation; only one patient had temporary facial paresis and salivary fistula. There was no tumor recurrence in both groups during the follow-up after 26 to 40 months) the endoscope-assisted parotidectomy group (median 30 months, range 26-40 months) and conventional group (median 32 months, range 26-40 months). CONCLUSION: Endoscope-assisted parotidectomy is a feasible technique. This procedure may serve as an alternative approach that allows a smaller incision. PMID- 19130966 TI - Assessment of the reproducibility of facial expressions with 3-D stereophotogrammetry. AB - OBJECTIVE: There is little research into analyzing whether facial expression changes are secondary to actual treatment or normal day-to-day variation. This study aims to ascertain whether nine-facial expressions were reproducible. SUBJECTS AND METHODS: Thirty-nine white subjects performed nine-facial expressions, captured by three-dimensional stereophotogrammetry in three sessions. After initial capture (session 1), each expression was repeated after 15 minutes (session 2) and then 4 weeks (session 3) after the initial session. Statistical analysis was performed on the mean variability of facial landmarks between session 1 and 2 and session 1 and 3. RESULTS: Repose was the most reproducible expression. The least reproducible was "blow-out-the-cheeks." Analysis between session 1 and 2 showed no significant differences in expression reproducibility. Analysis between session 1 and 3 showed significant differences for the "smile-with-lips-open" and "blow-out-the-cheeks" expressions. CONCLUSION: Facial expressions are reproducible in a 15-minute period. There are significant differences in the ability to reproduce facial expressions 4 weeks apart for "smile-with-lips-open" and "blow-out-the-cheeks." PMID- 19130967 TI - Ocular outcomes after gold weight placement and facial nerve resection. AB - OBJECTIVE: To assess the effectiveness of acute gold weight placement after facial nerve resection and to determine the role of concomitant lower eyelid procedures. STUDY DESIGN: Retrospective review. SUBJECTS AND METHODS: Twenty-two patients who received an upper eyelid gold weight at the time of parotidectomy and facial nerve resection were reviewed to assess ocular outcomes. RESULTS: After gold weight placement, twelve patients (12 of 22, 54.5%) subsequently presented with symptomatic ectropion (n = 9) and/or lagophthalmos (n = 5). Nine patients received a lower eyelid procedure (7 tarsal strips only, 1 tarsal strip combined with a lateral tarsorrhaphy, and 1 lateral tarsorrhaphy only). Six patients, in addition to a gold weight, also underwent a static sling to the midface at the time of facial nerve resection. None of these 6 received a subsequent lower eyelid procedure. Two patients required gold weight upsizing. Two patients required weight removal. CONCLUSIONS: Insertion of 1.2 gm upper eyelid weight with placement of midface sling is recommended at the time of facial nerve resection. Due to the need to tighten the lower eyelid in many of these patients, we now also consider performing a tarsal strip procedure at the time of facial nerve resection in any patient with pre-existing lower lid laxity. PMID- 19130968 TI - Sudden sensorineural hearing loss: an evaluation of treatment and management approaches by referring physicians. AB - OBJECTIVE: To assess and compare the current management of sudden sensorineural hearing loss (SSHL) between primary care physicians and general otolaryngologists. STUDY DESIGN: Written survey of physician practice patterns. METHODS: A multiple choice and Likert scale survey was mailed to 1306 otolaryngologists and primary care physicians in the upper midwest with respect to management of SSHL. RESULTS: A significant number of general practitioners treat SSHL independent of an otolaryngologist. General practitioners are significantly less impressed than otolaryngologists that steroids are an effective treatment (P < 0.0001). Over 98 percent of otolaryngologists use oral steroids as compared with 73 percent of general practitioners treating on their own. The vast majority of otolaryngologists start therapy with at least 60 mg of prednisone whereas lower doses and medrol dosepaks are more commonly used by general practitioners. Otolaryngologists are more likely to treat with steroids beyond one week of onset of hearing loss whereas general practitioners overwhelmingly will only treat within the first week. Approximately 50 percent of otolaryngologists add antiviral medications in contrast to 16 percent of generalists. CONCLUSION: The approach to SSHL differs between otolaryngologists and general practitioners. The lack of strong evidence-based guidelines for the treatment of SSHL may underlie the variability in management by first line providers. PMID- 19130969 TI - Levodopa does not enhance the effect of low-frequency repetitive transcranial magnetic stimulation in tinnitus treatment. AB - OBJECTIVE: Low-frequency repetitive transcranial magnetic stimulation (rTMS) has shown promise for the treatment of tinnitus. Experimental data from motor cortex stimulation in healthy subjects indicate that the suppressing effect of low frequency rTMS can be enhanced by dopaminergic receptor activation. Here we investigated whether administration of the dopamine precursor levodopa before low frequency rTMS enhances its efficacy in tinnitus treatment. STUDY DESIGN: Sixteen patients with chronic tinnitus received 100 mg of levodopa before each session of low-frequency rTMS. Results were compared with a matched control group of 16 patients who received the same treatment, but without levodopa. Treatment outcome was assessed with a standardized tinnitus questionnaire. RESULTS: Both stimulation protocols resulted in a significant reduction of tinnitus scores after 10 days of stimulation; however, there was no significant difference between the two groups. CONCLUSION: Our data suggest that 100 mg of levodopa does not enhance the effect of rTMS in the treatment of tinnitus. PMID- 19130970 TI - A minimally invasive registration method using surface template-assisted marker positioning (STAMP) for image-guided otologic surgery. AB - OBJECTIVE: A new, minimally invasive registration method was developed for image guided otologic surgery. We utilized laser-sintered template of the patient's bone surface to transfer the virtual markers to the patient's bone intraoperatively and eliminated the necessity for preoperative marker positioning or additional CT scan. STUDY DESIGN: Simulation surgeries and clinical application. SUBJECTS AND METHODS: We measured registration errors in 10 trials using replicas and six ear surgeries (two cochlear implant insertions, four translabyrinthine acoustic tumor removals). RESULTS: The target registration errors varied among the surgical targets. Errors were less than 1 mm near the cochlear implant insertion target both in phantom study and in actual surgeries. CONCLUSION: Our newly developed method reduced the preoperative procedures for patients but did not reduce the accuracy in cochlear implant surgery. Our method would be a useful image-guided surgery method in the field of otology, where both accuracy and noninvasiveness are required. PMID- 19130971 TI - Aminoglycoside ototoxicity in Nicaraguan children: patient risk factors and mitochondrial DNA results. AB - OBJECTIVE: Aminoglycoside ototoxicity remains a major problem in developing countries and accounts for 3 percent to 30 percent of hearing loss. This high prevalence rate may be related to genetic susceptibility from mitochondrial mutations in the 12S rRNA gene, comorbidity factors, or unregulated use of the medication. This study investigates the risk factors and prevalence of mtDNA mutations in serum from rural Nicaragua children with aminoglycoside-related hearing loss. STUDY DESIGN: Cross sectional study. SUBJECTS: Deaf children from rural Nicaragua. METHODS: Mitochondrial DNA isolated from serum collected from 31 deaf children with childhood or in utero exposure to gentamicin was amplified, sequenced, and analyzed for mutations in the 12S rRNA gene. RESULTS: No known pathologic mutations of the 12S rRNA gene were identified in this subpopulation of deaf children. In addition, patients with gentamicin exposure were often likely to have other comorbidity factors. CONCLUSION: These results suggest that genetic susceptibility is not a major factor in the high rate of gentamicin ototoxicity in this population sample. The high prevalence of gentamicin ototoxicty in this population is presumed to be due to unrestricted access to the drug. PMID- 19130972 TI - Influence of cochlear implantation on sacculus function. AB - OBJECTIVES: To examine the influence of cochlear implantation (CI) on the sacculus function and to analyze a possible correlation with resulting vertigo symptoms. METHODS: In a prospective study including 25 patients undergoing CI, preoperative and postoperative assessment of vestibular evoked myogenic potentials (VEMP), caloric horizontal semicircular canal (hSCC) response, and subjective vertigo symptoms were assessed. The patients with postoperative vertigo were compared to the patients without postoperative vertigo with regard to the findings of VEMP responses and caloric testing. RESULTS: Nine patients reported postoperative vertigo. Before surgery, eight of 23 patients (35%) had regular VEMP responses. Two months after CI, four patients had a new loss of sacculus function on VEMP testing. The CI represents a significant risk factor for sacculus impairment. In seven of 12 patients with preoperatively preserved caloric response, this decreased after the operation. The impaired vestibular function did not correlate with vertigo symptoms. When comparing the patient group with vertigo and the group without vertigo after CI, there is a significant difference in the patients' age. CONCLUSIONS: Although CI can cause damage to sacculus and hSCC function, this is probably not the only cause for postoperative vertigo. Advanced age is a significant risk factor for vertigo after CI. PMID- 19130973 TI - Receptive language outcomes in children after cochlear implantation. AB - OBJECTIVES: The objectives of the present study were (1) to assess receptive language scores in children after cochlear implantation and compare them with scores in normal hearing children and children with hearing loss that use hearing aids and (2) to determine how demographic factors, such as age of implantation, impact language outcomes. STUDY DESIGN: Case series. SUBJECTS/METHODS: Receptive language scores in children with profound prelingual hearing loss who received cochlear implants between 1996 and 2004 were analyzed. RESULTS: Standardized language assessments were available for 36 children. The average age at implantation was 33 months. The mean language scores for implanted children were within 1 standard deviation of scores of normal hearing individuals. Children with cochlear implants had significantly higher subtest scores (P < 0.05) than children with hearing aids. Children with additional disabilities had significantly (P < 0.05) poorer language performance. CONCLUSIONS: Pediatric cochlear implant recipients acquire receptive language skills that approach those of their hearing peers and exceed those of children with hearing aids. PMID- 19130974 TI - OK-432 sclerotherapy in head and neck lymphangiomas: long-term follow-up result. AB - INTRODUCTION: Nonsurgical treatments, such as sclerotherapy have been attempted for head and neck lymphagiomas. Of the available sclerosing agents, picibanil has shown satisfactory short-term treatment results in many studies, but no study has presented long-term treatment results. Accordingly, in the present study, the authors retrospectively reviewed the long-term treatment results of picibanil sclerotherapy. MATERIALS AND METHODS: Fifty-five lymphangioma patients who underwent picibanil sclerotherapy were enrolled. Data about initial and long-term response, recurrence, and excision rate were collected. RESULTS: Initial response rates were 83.5 percent and long-term response rates were 76.3 percent. CONCLUSION: Initial and the long-term response rate were equally good for lymphangioma. PMID- 19130975 TI - Transoral robotic assisted free flap reconstruction. PMID- 19130976 TI - Identification of the facial nerve main trunk by retrograde dissection of the postauricular branch. PMID- 19130977 TI - Giant salivary calculus of the submandibular gland. PMID- 19130978 TI - Isolated superior sternal cleft. PMID- 19130979 TI - Episodic painful parotid swelling caused by sialodochitis with eosinophilic inflammation: a new entity. PMID- 19130980 TI - FDG-positive Warthin's tumors in cervical lymph nodes mimicking metastases in tongue cancer staging with PET/CT. PMID- 19130981 TI - Chronic rhinosinusitis and persistent epiphora following conjunctivodacryocystorhinostomy. PMID- 19130982 TI - Recent advances in cardiopulmonary resuscitation: cardiocerebral resuscitation. AB - Cardiocerebral resuscitation (CCR) is a new approach for resuscitation of patients with cardiac arrest. It is composed of 3 components: 1) continuous chest compressions for bystander resuscitation; 2) a new emergency medical services (EMS) algorithm; and 3) aggressive post-resuscitation care. The first 2 components of CCR were first instituted in 2003 in Tucson, Arizona; in 2004 in the Rock and Walworth counties of Wisconsin; and in 2005 in the Phoenix, Arizona, metropolitan area. The CCR method has been shown to dramatically improve survival in the subset of patients most likely to survive: those with witnessed arrest and shockable rhythm on arrival of EMS. The CCR method advocates continuous chest compressions without mouth-to-mouth ventilations for witnessed cardiac arrest. It advocates either prompt or delayed defibrillation, based on the 3-phase time sensitive model of ventricular fibrillation (VF) articulated by Weisfeldt and Becker. For bystanders with access to automated external defibrillators and EMS personnel who arrive during the electrical phase (i.e., the first 4 or 5 min of VF arrest), the delivery of prompt defibrillator shock is recommended. However, EMS personnel most often arrive after the electrical phase -- in the circulatory phase of VF arrest. During the circulatory phase of VF arrest, the fibrillating myocardium has used up much of its energy stores, and chest compressions that perfuse the heart are mandatory prior to and immediately after a defibrillator shock. Endotracheal intubation is delayed, excessive ventilations are avoided, and early-administration epinephrine is advocated. PMID- 19130983 TI - Cardiocerebral resuscitation: a broader perspective. AB - Historically, cardiac arrest outcomes have been stagnant with few therapies demonstrating clinical benefit. Recent advances in our understanding of cardiac arrest physiology and therapy have led to improved outcomes and renewed interest in defining the "optimal" approach. Cardiocerebral resuscitation (CCR) represents a bundle of specific therapies designed to enhance perfusion during cardiopulmonary arrest by emphasizing chest compressions over ventilations and "priming" the heart with compressions before and after defibrillation attempts. Implemented in Arizona and Wisconsin in 2003, patients treated using CCR appear to have improved outcomes compared with those treated under the 2000 guidelines from the International Liaison Committee on Resuscitation (ILCOR). This was particularly true in the subgroup of patients with bystander-witnessed collapse, who may represent a group with adequate oxygen reserves at the time of arrest and decreased requirement for immediate positive-pressure ventilation. Many components of CCR have since been incorporated in the 2005 ILCOR guidelines. Beyond the specific treatment approaches that define CCR, this alternative approach may represent the future of resuscitation science in which each institution and emergency medical services agency will define an optimal approach to treatment and training based on the specific resources available and patient population. This may mandate a paradigm shift away from advanced cardiac life support and basic life support, which emphasize standardization of content and format rather than institution- or agency-specific protocols and training. PMID- 19130984 TI - Utilization and impact of pre-hospital electrocardiograms for patients with acute ST-segment elevation myocardial infarction: data from the NCDR (National Cardiovascular Data Registry) ACTION (Acute Coronary Treatment and Intervention Outcomes Network) Registry. AB - OBJECTIVES: This study sought to determine the association of pre-hospital electrocardiograms (ECGs) and the timing of reperfusion therapy for patients with ST-segment elevation myocardial infarction (STEMI). BACKGROUND: Pre-hospital ECGs have been recommended in the management of patients with chest pain transported by emergency medical services (EMS). METHODS: We evaluated patients with STEMI from the NCDR (National Cardiovascular Data Registry) ACTION (Acute Coronary Treatment and Intervention Outcomes Network) registry who were transported by EMS from January 1, 2007, through December 31, 2007. Patients were stratified by the use of pre-hospital ECGs, and timing of reperfusion therapy was compared between the 2 groups. RESULTS: A total of 7,098 of 12,097 patients (58.7%) utilized EMS, and 1,941 of these 7,098 EMS transport patients (27.4%) received a pre-hospital ECG. Among the EMS transport population, primary percutaneous coronary intervention was performed in 92.1% of patients with a pre-hospital ECG versus 86.3% with an in-hospital ECG, whereas fibrinolytic therapy was used in 4.6% versus 4.2% of patients. Median door-to-needle times for patients receiving fibrinolytic therapy (19 min vs. 29 min, p = 0.003) and median door-to-balloon times for patients undergoing primary percutaneous coronary intervention (61 min vs. 75 min, p < 0.0001) were significantly shorter for patients with a pre hospital ECG. A suggestive trend for a lower risk of in-hospital mortality was observed with pre-hospital ECG use (adjusted odds ratio: 0.80, 95% confidence interval: 0.63 to 1.01). CONCLUSIONS: Only one-quarter of these patients transported by EMS receive a pre-hospital ECG. The use of a pre-hospital ECG was associated with a greater use of reperfusion therapy, faster reperfusion times, and a suggested trend for a lower risk of mortality. PMID- 19130985 TI - Leptin and coronary heart disease: prospective study and systematic review. AB - OBJECTIVES: This study sought to better determine the link between leptin and coronary heart disease (CHD). BACKGROUND: Circulating leptin is considered a risk factor for CHD but larger studies are needed. METHODS: Leptin levels were measured in 550 men with fatal CHD or nonfatal myocardial infarction and in 1,184 controls nested within a prospective study of 5,661 British men and set in context with a meta-analysis. RESULTS: Baseline leptin correlated with body mass index (BMI), blood pressure, total cholesterol, triglyceride, and inflammatory markers; correlations persisted after BMI adjustment. The within-person consistency of leptin values over 4 years (correlation coefficient: 0.79; 95% confidence interval [CI]: 0.73 to 0.83) was higher than those of some established cardiovascular risk factors. In a comparison of individuals in the top third with those in the bottom third of baseline leptin, the age- and town-adjusted odds ratio for CHD was 1.25 (95% CI: 0.96 to 1.62), decreasing to 0.98 (95% CI: 0.72 to 1.34) after adjustment for BMI. A systematic review identified 7 prospective reports with heterogeneous findings (I(2) = 60%, 13% to 82%). The combined adjusted risk ratio across all studies was 1.44 (95% CI: 0.95 to 2.16) in a comparison of extreme thirds of leptin levels. The inconsistency between studies was partially explained by sample size, with combined estimates from studies involving >100 CHD cases (1.28, 95% CI: 0.80 to 2.04) being somewhat weaker than those from smaller studies (1.81, 95% CI: 0.76 to 4.31). CONCLUSIONS: Previous studies appear to have overestimated associations of leptin and CHD risk. Our results suggest a moderate association that is largely dependent on BMI. PMID- 19130986 TI - Depression and cardiovascular health care costs among women with suspected myocardial ischemia: prospective results from the WISE (Women's Ischemia Syndrome Evaluation) Study. AB - OBJECTIVES: This study evaluated 3 novel questions in a prospective clinical cohort of women undergoing evaluation for suspected myocardial ischemia: 1) What is the relationship between depression and cardiovascular costs? 2) Does the relationship vary by definition of depression? 3) Do depression-cost relationship patterns differ among women with versus without coronary artery disease (CAD)? BACKGROUND: Comorbid depression has been linked to higher medical costs in previous studies of cardiovascular patients. METHODS: A total of 868 women presenting with suspected myocardial ischemia completed an extensive baseline examination including cardiovascular risk factor assessment and coronary angiogram. Depression was defined by: 1) current use of antidepressants; 2) a reported history of depression treatment; and 3) Beck Depression Inventory scores. Direct (hospitalizations, office visits, procedures, and medications) and indirect (out-of-pocket, lost productivity, and travel) costs were collected through 5 years of follow-up to estimate cardiovascular costs. RESULTS: Using the study criteria, 17% to 45% of the women studied met study depression criteria. Depressed women showed adjusted annual cardiovascular costs $1,550 to $3,300 higher than nondepressed groups (r = 0.08 to 0.12, p < 0.05). Depression-cost relationships also varied by CAD status, with stronger associations present among women without evidence of significant CAD. CONCLUSIONS: Depression was associated with 15% to 53% increases in 5-year cardiovascular costs, and cost differences were present using 3 definitions of depression. The results reinforce the importance of assessing depression in clinical populations and support the hypothesis that improved management of depression in women with suspected myocardial ischemia could reduce medical costs. PMID- 19130987 TI - Clinical effectiveness of beta-blockers in heart failure: findings from the OPTIMIZE-HF (Organized Program to Initiate Lifesaving Treatment in Hospitalized Patients with Heart Failure) Registry. AB - OBJECTIVES: We sought to examine associations between initiation of beta-blocker therapy and outcomes among elderly patients hospitalized for heart failure. BACKGROUND: Beta-blockers are guideline-recommended therapy for heart failure, but their clinical effectiveness is not well understood, especially in elderly patients. METHODS: We merged Medicare claims data with OPTIMIZE-HF (Organized Program to Initiate Lifesaving Treatment in Hospitalized Patients With Heart Failure) records to examine long-term outcomes of eligible patients newly initiated on beta-blocker therapy. We used inverse probability-weighted Cox proportional hazards models to determine the relationships among treatment and mortality, rehospitalization, and a combined mortality-rehospitalization end point. RESULTS: Observed 1-year mortality was 33%, and all-cause rehospitalization was 64%. Among 7,154 patients hospitalized with heart failure and eligible for beta-blockers, 3,421 (49%) were newly initiated on beta-blocker therapy. Among patients with left ventricular systolic dysfunction (LVSD) (n = 3,001), beta-blockers were associated with adjusted hazard ratios of 0.77 (95% confidence interval [CI]: 0.68 to 0.87) for mortality, 0.89 (95% CI: 0.80 to 0.99) for rehospitalization, and 0.87 (95% CI: 0.79 to 0.96) for mortality rehospitalization. Among patients with preserved systolic function (n = 4,153), beta-blockers were associated with adjusted hazard ratios of 0.94 (95% CI: 0.84 to 1.07) for mortality, 0.98 (95% CI: 0.90 to 1.06) for rehospitalization, and 0.98 (95% CI: 0.91 to 1.06) for mortality-rehospitalization. CONCLUSIONS: In elderly patients hospitalized with heart failure and LVSD, incident beta-blocker use was clinically effective and independently associated with lower risks of death and rehospitalization. Patients with preserved systolic function had poor outcomes, and beta-blockers did not significantly influence the mortality and rehospitalization risks for these patients. PMID- 19130988 TI - Prognostic implications of baroreflex sensitivity in heart failure patients in the beta-blocking era. AB - OBJECTIVES: This study investigated the clinical correlates and prognostic value of depressed baroreceptor-heart rate reflex sensitivity (BRS) among patients with heart failure (HF), with and without beta-blockade. BACKGROUND: Abnormalities in autonomic reflexes play an important role in the development and progression of HF. Few studies have assessed the effects of beta-blockers on BRS in HF. METHODS: The study population consisted of 103 stable HF patients, age (median [interquartile range]) 54 years (48 to 57 years), with New York Heart Association (NYHA) functional class > or =III in 22, and with a left ventricular ejection fraction (LVEF) of 30% (24% to 36%), treated with beta-blockers; and 144 untreated patients, age 55 years (48 to 60 years), with NYHA functional class > or =III in 47%, and an LVEF of 26% (21% to 30%). They underwent BRS testing (phenylephrine technique). RESULTS: In both treated and untreated patients, a lower BRS was associated with a higher (> or =III) NYHA functional class (p = 0.0002 and p < 0.0001, respectively); a more severe (> or =2) mitral regurgitation (p = 0.007 and p = 0.0002), respectively; a lower LVEF (p = 0.0004 and p = 0.001, respectively), baseline RR interval (p = 0.0004 and p = 0.0002, respectively), and SDNN (p < 0.0001, p = 0.002, respectively); and a higher blood urea nitrogen (p = 0.004, p < 0.0001, respectively). Clinical variables explained only 43% of BRS variability among treated and 36% among untreated patients. During a median follow-up of 29 months, 17 of 103 patients and 55 of 144 patients, respectively, experienced a cardiac event. A depressed BRS (<3.0 ms/mm Hg) was significantly associated with the outcome, independently of known risk predictors and beta-blocker treatment (adjusted hazard ratio: 3.0 [95% confidence interval: 1.5 to 5.9], p = 0.001). CONCLUSIONS: Baroreceptor-heart rate reflex sensitivity does not simply mirror the pathophysiological substrate of HF. A depressed BRS conveys independent prognostic information that is not affected by the modification of autonomic dysfunction brought about by beta-blockade. PMID- 19130989 TI - Endothelial dysfunction induced by post-prandial lipemia: complete protection afforded by high-intensity aerobic interval exercise. AB - OBJECTIVES: This study was designed to study the effect of exercise and a high fat meal (HFM) on endothelial function. BACKGROUND: Post-prandial lipemia and exercise oppose each other in terms of cardiovascular risk; however, the mechanism of their interaction is not well understood. METHODS: Endothelial function was assessed by brachial artery flow-mediated dilation (FMD) in 8 healthy men before and after an HFM preceded (16 to 18 h) by rest, a single bout of continuous moderate-intensity exercise (CME), and high-intensity interval exercise (HIIE). RESULTS: Before the HFM, initial brachial artery diameters were similar in all trials (0.43 +/- 0.04 cm), but after the HFM, basal diameter decreased only in the control (0.39 +/- 0.03 cm) and CME (0.38 +/- 0.04 cm) trials. Before the HFM, FMD/shear was improved by a single bout of CME (+20%, p < 0.01) and HIIE (+45%, p < 0.01; group differences, p < 0.01), with no effect in the control trial. After the HFM (30, 120, and 240 min), FMD decayed to a lesser extent with CME, but in a similar fashion to the control trial. In contrast, FMD in the HIIE trial remained elevated following the exercise despite a clear meal induced lipemia. Although there were no correlations between vascular function and food-induced markers of cardiovascular risk, antioxidant status was strongly correlated with FMD (r = 0.9, p < 0.001). CONCLUSIONS: These findings reveal a clinically relevant protective effect of acute exercise on the vasculature that is clearly exercise intensity dependent and tightly related to exercise-induced antioxidant capacity. (Endothelial Dysfunction Induced by Postprandial Lipemia; NCT00660491). PMID- 19130991 TI - Myocardial effects of PDE5 inhibition: more function with less mass. PMID- 19130990 TI - Sildenafil stops progressive chamber, cellular, and molecular remodeling and improves calcium handling and function in hearts with pre-existing advanced hypertrophy caused by pressure overload. AB - OBJECTIVE: This study sought to test the efficacy of phosphodiesterase type 5A (PDE5A) inhibition for treating advanced hypertrophy/remodeling caused by pressure overload, and to elucidate cellular and molecular mechanisms for this response. BACKGROUND: Sildenafil (SIL) inhibits cyclic guanosine monophosphate specific PDE5A and can blunt the evolution of cardiac hypertrophy and dysfunction in mice subjected to pressure overload. Whether and how it ameliorates more established advanced disease and dysfunction is unknown. METHODS: Mice were subjected to transverse aortic constriction (TAC) for 3 weeks to establish hypertrophy/dilation, and subsequently treated with SIL (100 mg/kg/day) or placebo for 6 weeks of additional TAC. RESULTS: The SIL arrested further progressive chamber dilation, dysfunction, fibrosis, and molecular remodeling, increasing myocardial protein kinase G activity. Isolated myocytes from TAC-SIL hearts showed greater sarcomere shortening and relaxation, and enhanced Ca(2+) transients and decay compared with nontreated TAC hearts. The SIL treatment restored gene and protein expression of sarcoplasmic reticulum Ca(2+) uptake adenosine triphosphatase (SERCA2a), phospholamban (PLB), and increased PLB phosphorylation (S16), consistent with improved calcium handling. The phosphatase calcineurin (Cn) and/or protein kinase C-alpha (PKCalpha) can both lower phosphorylated phospholamban and depress myocyte calcium cycling. The Cn expression and PKCalpha activation (outer membrane translocation) were enhanced by chronic TAC and reduced by SIL treatment. Expression of PKCdelta and PKCepsilon also increased with TAC but were unaltered by SIL treatment. CONCLUSIONS: SIL treatment applied to well-established hypertrophic cardiac disease can prevent further cardiac and myocyte dysfunction and progressive remodeling. This is associated with improved calcium cycling, and reduction of Cn and PKCalpha activation may be important to this improvement. PMID- 19130992 TI - Polymorphic ventricular tachycardia from R-on-T pacing. PMID- 19130993 TI - Is it reasonable to treat all calcified stenotic valves with a valve stent? Probably yes if we get a full stent expansion. PMID- 19130995 TI - President's message: training in echocardiography in the multimodality era. PMID- 19130996 TI - Sonographers' communication: expanding the role of the cardiac sonographer. PMID- 19130997 TI - Intraoperative three-dimensional echocardiography: ready for primetime? PMID- 19130998 TI - Echocardiographic assessment of valve stenosis: EAE/ASE recommendations for clinical practice. PMID- 19130999 TI - Role of echocardiography in the diagnosis of constrictive pericarditis. AB - The clinical recognition of constrictive pericarditis (CP) is important but challenging. In addition to Doppler echocardiography, newer echocardiographic techniques for deciphering myocardial deformation have facilitated the noninvasive recognition of CP and its differentiation from restrictive cardiomyopathy. In a patient with heart failure and a normal ejection fraction, echocardiographic demonstration of exaggerated interventricular interdependence, relatively preserved left ventricular longitudinal deformation, and attenuated circumferential deformation is diagnostic of CP. This review is a concise update on the pathophysiology and hemodynamic features of CP, the transmural and torsional mechanics of CP, and the merits and pitfalls of the various echocardiographic techniques used in the diagnosis of CP. PMID- 19131000 TI - Real-time three-dimensional transesophageal echocardiography in the intraoperative assessment of mitral valve disease. AB - BACKGROUND: The aims of this study were to evaluate the feasibility of real-time 3-dimensional (3D) transesophageal echocardiography in the intraoperative assessment of mitral valve (MV) pathology and to compare this novel technique with 2-dimensional (2D) transesophageal echocardiography. METHODS: Forty-two consecutive patients undergoing MV repair for mitral regurgitation (MR) were studied prospectively. Intraoperative 2D and 3D transesophageal echocardiographic (TEE) examinations were performed using a recently introduced TEE probe that provides real-time 3D imaging. Expert echocardiographers blinded to 2D TEE findings assessed the etiology of MR on 3D transesophageal echocardiography. Similarly, experts blinded to 3D TEE findings assessed 2D TEE findings. Both were compared with the anatomic findings reported by the surgeon. RESULTS: At the time of surgical inspection, ischemic MR was identified in 12% of patients, complex bileaflet myxomatous disease in 31%, and specific scallop disease in 55%. Three dimensional TEE image acquisition was performed in a short period of time (60 +/- 18 seconds) and was feasible in all patients, with optimal (36%) or good (33%) imaging quality in the majority of cases. Three-dimensional TEE imaging was superior to 2D TEE imaging in the diagnosis of P1, A2, A3, and bileaflet disease (P < .05). CONCLUSIONS: Real-time 3D transesophageal echocardiography is a feasible method for identifying specific MV pathology in the setting of complex disease and can be expeditiously used in the intraoperative evaluation of patients undergoing MV repair. PMID- 19131001 TI - Impact of cardiac rhythm on mitral valve area calculated by the pressure half time method in patients with moderate or severe mitral stenosis. AB - BACKGROUND: The pressure half-time (PHT) method has been widely used to estimate mitral valve area (MVA) in patients with mitral stenosis (MS), in the belief that this simple method provides reliable information on true MVA. However, its limitation has been repeatedly recognized under different circumstances. The aim of this study was to evaluate the effect of cardiac rhythm on PHT-derived MVA calculation in relation to net atrioventricular compliance (C(n)). METHODS: Patients (n = 41) with rheumatic pure moderate or severe MS were consecutively recruited. Eighteen patients with sinus rhythm were allocated to group 1 and the remaining patients with chronic atrial fibrillation to group 2. MVA was obtained using the PHT method and by planimetry (considered the gold standard in this study). C(n) was calculated with a previously validated equation. RESULTS: There were no differences between the 2 groups in terms of age, gender, left ventricular dimensions or ejection fraction, or transmitral pressure gradient. Left atrial volume index (134.6 +/- 106.7 vs 79.2 +/- 16.8 mL; P = .003) and C(n) (6.6 +/- 1.5 vs 4.7 +/- 1.2 mL/mm Hg; P < .001) were higher in group 2 than in group 1. Disagreement of MVA estimation by PHT compared with that by 2 dimensional planimetry was 8.0 +/- 19.2% for group 1 and -24.9 +/- 13.9% for group 2 (P = .002). In addition, the extent of disagreement of PHT-derived MVA compared with that by 2-dimensional planimetry was significantly correlated with C(n) (r(2) = 0.71, P < .001). MVA by the PHT method was estimated to be substantially higher in patients with C(n) values < 4 mL/mm Hg, most of whom were in sinus rhythm. Multivariate analysis confirmed the independent association of cardiac rhythm with discrepancy of PHT-derived MVA compared with that by planimetry. CONCLUSION: Changes in cardiac rhythm with associated modifications of C(n) can alter the accuracy of the PHT method for estimating MVA. Given the limitation described here, 2-dimensional planimetry, not the PHT method, should be used as a primary echocardiographic tool for MVA calculation. PMID- 19131003 TI - Appropriateness criteria for echocardiography: an important step toward improving quality. PMID- 19131002 TI - Application of appropriateness criteria in outpatient transthoracic echocardiography. AB - BACKGROUND: Appropriateness criteria were applied to outpatient transthoracic echocardiographic (TTE) studies. METHODS: Indications were rated as appropriate, inappropriate, or unclassifiable, considering provider-stated indications, previous TTE studies, symptom changes, and patient-stated indications. Clinically important new or unexpected findings were recorded. RESULTS: Of 368 TTE studies, 206 (56%) were appropriate, 31 (8%) were inappropriate, and 131 (35%) were unclassifiable. Appropriateness was not correlated with patient or provider demographics. In 288 cases with prior TTE studies, there were 92 (32%) important new findings and 63 (22%) unexpected findings, of which 20% were from inappropriately ordered and 31% from unclassifiable TTE studies. Appropriateness was not associated with new (odds ratio, 1.23; 95% confidence interval, 0.48 3.18) or unexpected (odds ratio, 1.15; 95% confidence interval, 0.38-3.52) findings. Provider type and level of training were not correlated with new or unexpected findings. CONCLUSIONS: Many indications for TTE studies were unclassifiable. A high percentage of inappropriately ordered TTE studies yielded important information. Care must be taken in judging the value of TTE studies solely on the basis of appropriateness criteria. PMID- 19131004 TI - Automated assessment of left atrial function from time-left atrial volume curves using a novel speckle tracking imaging method. AB - BACKGROUND: Using a novel speckle-tracking imaging method, time-left atrial (LA) volume curves (TLAVCs) can be automatically obtained. The aim of this study was to evaluate whether this method can be used for the measurement of LA function with TLAVCs. METHODS: In 10 normal subjects and 20 patients, apical 4-chamber images were obtained. Maximum volume, reservoir volume, conduit volume, booster pump volume, and minimum volume were measured from TLAVCs. The results were compared with those obtained by the manual tracing method on every frame during 1 cardiac cycle. RESULTS: There was good agreement between the speckle-tracking imaging and manual methods for maximum LA volume (r = 0.98, P < .001), reservoir volume (r = 0.82, P < .001), conduit volume (r = 0.87, P < .001), booster pump volume (r = 0.80, P < .001), and minimum volume (r = 0.98, P < .001). The time to obtain TLAVCs was significantly shorter with the speckle-tracking imaging method (64 +/- 22 seconds) than with the manual method (22 +/- 4 minutes). CONCLUSION: TLAVCs obtained using the speckle-tracking imaging method can be used for the rapid and noninvasive automated quantitation of LA function. PMID- 19131005 TI - Effects of aging on left atrial function assessed by two-dimensional speckle tracking echocardiography. AB - BACKGROUND: Aging affects left atrial (LA) function, which can be assessed by two dimensional (2D) speckle tracking echocardiography (STE). The aim of this study was to determine (1) the feasibility and accuracy of measuring LA volume with 2DSTE and (2) the effects of aging on LA function. METHODS: 2DSTE of the LA was acquired from the apical 4-chamber view (frame rate: 63 +/- 11 /sec, iE33) using prototype speckle tracking software (QLAB, Philips Medical Systems, Andover, MA) in 140 healthy volunteers (3-79 years, 74 men). LA wall was tracked on a frame-by frame basis, and LA volume waveforms were generated. Maximum LA volume (LAVmax) and minimal LA volume (LAVmin), and the LA volume before atrial contraction (LAVpre-a) were measured. Passive emptying percent of total emptying (LA conduit function) and active emptying percent of total emptying (booster function) were calculated as ([LAVmax-LAVpre-a]/[LAVmax-LAVmin]) x 100 and ([LAVa LAVmin]/[LAVmax-LAVmin]) x 100. RESULTS: Adequate LA volume waveforms were obtained in all subjects. A good correlation was obtained between speckle tracking-derived LA volume measurements and manually traced LA volume measurements of the identical 2D image (LAVmax: r = 0.93, P <.001, LAVmin: r = 0.88, P <.001, LAVpre-a: r = 0.92, P <.001). Passive and active emptying indices had a significant age dependency (r = 0.80, P <.001). Overall, passive emptying accounted for 67% of the total LA emptying ranging from 83% in the youngest to 42% in the oldest decade. CONCLUSION: Aging significantly affects LA conduit and booster function. 2DSTE can effectively and easily measure LA volume and has a potential for the noninvasive assessment of LA function. PMID- 19131006 TI - Prognostic significance of left atrial volume dilatation in patients with hypertrophic cardiomyopathy. AB - To evaluate the prognostic role of left atrial (LA) volume in hypertrophic cardiomyopathy (HCM), LA volume was measured at baseline and during follow-up in 140 patients with HCM. Unfavorable outcome, defined as occurrence of sudden death, heart transplantation, or invasive reduction of obstruction, developed in 16 patients. In patients with enlarged LA volume (>27 mL/m(2)), there was an increased risk for unfavorable outcome (P = .0152). Patients with normal LA volume at baseline in whom volume increased more than 3 mL per year (fast dilating LA volume) had a worse prognosis than patients with normal and stable volume (P < .001) and similar to patients with dilated LA volume at baseline (P = not significant). LA volume dilated at baseline, fast dilating LA volume, and New York Heart Association functional class were independent predictors of unfavorable outcome development (odds ratio: 11.453; P = .021, P = 2.019, P = .020, respectively). The assessment of LA volume at baseline and during follow-up adds information regarding prognosis in patients with HCM. PMID- 19131007 TI - Left atrial volume provides independent prognostic value in patients with Chagas cardiomyopathy. AB - BACKGROUND: Heart failure is a severe clinical manifestation of Chagas disease. Left atrial volume (LAV), a marker of chronically elevated left ventricular (LV) filling pressure, is a predictor of prognosis in patients with heart failure and may be important in the assessment of risk in patients with Chagas disease. The aim of this study was to identify echocardiographic parameters of diastolic function predictors of survival in patients with Chagas cardiomyopathy. METHODS: A total of 192 patients with Chagas cardiomyopathy (mean age, 48.5 +/- 12.1 years; 37% women) were prospectively enrolled. The end points were death and cardiac transplantation. RESULTS: Over a mean follow-up period of 33.8 months, LAV normalized for body surface area emerged as an independent predictor of survival (hazard ratio, 1.037 per 1 mL/m(2) change; 95% confidence interval, 1.018-1.056; P < .001), adding incremental prognostic value to clinical factors, LV ejection fraction, and Doppler-derived parameters of diastolic function. Receiver operating characteristic curve analysis identified the best cutoff values for the prediction of end points. LAV > 51 mL/m(2) was associated with significant excess mortality (log-rank, P < .001). CONCLUSIONS: LAV provides powerful prognostic information incrementally and independently to clinical data and conventional echocardiographic parameters in the prediction of survival. New York Heart Association functional class, LV ejection fraction, right ventricular function, the E/E' ratio, and LAV index can be used to build a risk prediction model, which can be used clinically. PMID- 19131009 TI - A New Year, a new look. PMID- 19131008 TI - Left ventricular twist mechanics in a canine model of reversible congestive heart failure: a pilot study. AB - BACKGROUND: Left ventricular (LV) twist dynamics play an important role in LV systolic and diastolic function. The aim of this preliminary study was to investigate LV twist dynamics in a canine model of reversible congestive heart failure (CHF). METHODS: Pacing systems were implanted in adult dogs, and continuous chronic right ventricular pacing (230-250 beats/min) was applied until CHF induction. Pacing was then stopped to allow the heart to recover. Echocardiography and LV catheterization were performed at baseline, during CHF while pacing was temporarily switched off, and during recovery. LV twist was computed as the difference between apical and basal rotation measured using 2 dimensional speckle tracking. Torsion was further calculated as LV twist divided by the LV long axis. The untwisting rate was computed as the peak diastolic time derivative of twist. RESULTS: In 6 dogs that completed the study, we found that CHF developed after 2 to 4 weeks of pacing, with LV end-diastolic volume, end systolic volume, end-diastolic pressure, and the time constant of relaxation during isovolumic relaxation period (tau) all increasing significantly compared with baseline and recovering to normal levels 2 to 4 weeks after pacing was stopped. LV twist, torsion, and untwisting rate decreased significantly with CHF compared with baseline and improved during recovery from CHF. CONCLUSION: LV twist dynamics reflect pacing-induced CHF and its reversal as assessed by echocardiographic speckle tracking. PMID- 19131010 TI - Retained fibrin sleeve: transesophageal echocardiographic observations. AB - The role of echocardiography for the evaluation of thrombus formation on indwelling intracardiac catheters is well established. Considerably less well described, however, are the echocardiographic characteristics of the so-called retained fibrin sheath, a sleeve of fibrin that surrounds the catheter at the point at which it enters the vein that commonly remains adherent to the vessel wall after catheter removal. The authors report the transesophageal echocardiographic findings of a retained fibrin sheath following catheter removal in a patient with end-stage renal disease and infective endocarditis of the aortic valve. PMID- 19131011 TI - Witnessed massive pulmonary thromboembolism and dynamic left ventricular outflow tract obstruction. AB - The authors describe a patient with a floating thrombus in the right heart that migrated into the pulmonary artery, causing on-site massive pulmonary thromboembolism during echocardiography and systolic anterior motion of the anterior mitral leaflet resulting in significant dynamic left ventricular outflow tract obstruction. Reduced pulmonary circulation due to pulmonary embolism and decreased systemic circulation associated with left ventricular outflow tract obstruction resulted in a fatal outcome. PMID- 19131012 TI - Abnormally thickened papillary muscle resulting in dynamic left ventricular outflow tract obstruction: an unusual presentation of hypertrophic cardiomyopathy. AB - We report the case of a symptomatic 18-year-old patient, gene-positive for hypertrophic cardiomyopathy (HCM), who presented with symptomatic dynamic left ventricular outflow tract (LVOT) obstruction caused by an abnormally thickened papillary muscle in the absence of septal hypertrophy. This was confirmed using multimodality imaging, including echocardiography and magnetic resonance imaging. He successfully underwent surgery for papillary muscle realignment without septal myectomy. PMID- 19131013 TI - Synthesis of observational studies should consider credibility ceilings. AB - OBJECTIVE: Meta-analyses of observational studies often get spuriously precise results. We aimed to factor this skepticism in meta-analysis calculations. STUDY DESIGN AND SETTING: We developed a simple sensitivity analysis starting from the assumption that any single observational study cannot give us more than a maximum certainty c% (called credibility ceiling) that an effect is in a particular direction and not in the other. Each study included in meta-analysis is adjusted for different credibility ceilings c and the consistency of the conclusion examined. We applied the method in three meta-analyses of observational studies with nominally statistically significant summary effects (mortality with teaching versus nonteaching health care; risk of non-Hodgkin's lymphoma with hair dyes; mortality with omega-3 fatty acids). RESULTS: Between-study heterogeneity I(2) estimates dropped from 36%-72% without a ceiling effect to 0% with ceilings of 9%, 4%, and 4% in the three meta-analyses, respectively. Nominal statistical significance was lost with ceilings of 10%, 8%, and 11%, respectively. The likelihood ratios suggested that even with minimal ceiling effects, there was no strong support for the credibility of each of these three associations. CONCLUSIONS: Consideration of credibility ceilings allows conservative interpretation of observational evidence and can be applied routinely to meta analyses of observational studies. PMID- 19131014 TI - Spuriously precise results from meta-analysis. Is better statistical correction or a more critical methodological assessment warranted? PMID- 19131016 TI - Money talks: non-monetary incentive and Internet administration fail to increase response rates to a physician survey. PMID- 19131018 TI - Supraventricular tachycardia in a patient with a ruptured abdominal aortic aneurysm. PMID- 19131019 TI - Recent articles of interest to critical care out-of-hospital personnel. PMID- 19131020 TI - Fixing what's broken. PMID- 19131021 TI - LifeFlight of Maine: the gift of trust. PMID- 19131022 TI - The changing role of air medical communication specialists. PMID- 19131023 TI - Partners in care: implementing a policy on family member passengers. PMID- 19131024 TI - Transporting the pregnant patient in shock: case report and review. PMID- 19131026 TI - Veterinary clinics of North America. Exotic animal practice. Cardiology. Preface. PMID- 19131025 TI - Transfer of patients dependent on an intra-aortic balloon pump using critical care services. AB - INTRODUCTION: The intra-aortic balloon pump (IABP) is a hemodynamic support device that provides circulatory enhancement to patients whose cardiac output is compromised. Special clinical skills are required for management of the patient with an IABP in place. Few studies have discussed the transport of the IABP dependent patient. The current study was designed to describe the transport of IABP-dependent patients, with a focus on pretransport interventions, transport interventions, quality improvement, and complications. METHODS: A review of all transports from January 1, 2004, through December 31, 2005, performed by a critical care transport program with a nurse/paramedic crew offering mobile intensive care unit (ICU), rotor-wing, and fixed-wing service was conducted. All patients who were maintained on an intra-aortic balloon pump (IABP) were eligible for inclusion. A certified perfusionist was available for consultation on all transports. Information about the IABP, including the pump timing, confirmation of balloon location, and inflation/deflation timing parameters, was collected. Proper balloon placement was verified and recorded at the sending hospital. Data were collected regarding interventions required before and during transport and complications during transport. Descriptive statistics were used. RESULTS: During the study period, 173 transports involving an IABP were performed. The average age was 60.8 years, and 67.8% were men. Forty-one percent were flown by rotor wing, 36.4% were transported by the mobile ICU, and 21.4% were flown by the fixed wing transport. In 1.2% of cases, there was a change in transport mode. Twelve percent of patients required some increase in oxygen supplementation, but only one patient required intubation before transport by the transport crew. The most common pretransport medications were heparin (69%), inotropes (55%), and other infusions (46.8%). Twenty-two percent had no written confirmation of the correct balloon placement. There were no significant complications found during transport, including hemorrhage, loss of trigger signals, or cardiac arrest. Twelve percent had some abnormalities in timing of balloon inflation or deflation. CONCLUSION: IABP transports can be safely performed by a nurse/paramedic critical care transport team with perfusionist consultation. Few patients require significant intervention before transport. Attention must be paid to balloon inflation and deflation timing despite the existence of timing algorithms. Significant complications during transport were not seen. Future studies should explore the overall outcome of IABP-dependent patients and the role of transport mode on outcome. PMID- 19131027 TI - Cardiac biology and disease in invertebrates. AB - It may seem ridiculous to consider cardiac diseases in sub-vertebrate animals; when on earth is a tarantula, a butterfly or a snail going to be presented as a clinical case with heart failure or a congenital cardiac abnormality? This article examines the work of research groups investigating invertebrates as valuable models of heart disease in man. Examining invertebrates with gene defects similar to those in human patients with heart disease, congenital or acquired, allows us to probe deeply into the aetiopathogenesis of many cardiac conditions. PMID- 19131028 TI - Fish cardiovascular physiology and disease. AB - Fish patients with cardiovascular disorders present a challenge in terms of diagnostic evaluation and therapeutic options. Veterinarians can approach these cases in fish using methods similar to those employed for other companion animals. Clinicians who evaluate and treat fish in private, aquarium, zoologic, or aquaculture settings need to rely on sound clinical judgment after thorough historical and physical evaluation. Pharmacokinetic data and treatments specific to cardiovascular disease in fish are limited; thus, drug types and dosages used in fish are largely empiric. Fish cardiovascular anatomy, physiology, diagnostic evaluation, monitoring, common diseases, cardiac pathologic conditions, formulary options, and comprehensive references are presented with the goal of providing fish veterinarians with clinically relevant tools. PMID- 19131029 TI - Cardiovascular physiology and diseases of amphibians. AB - The class Amphibia includes three orders of amphibians: the anurans (frogs and toads), urodeles (salamanders, axolotls, and newts), and caecilians. The diversity of lifestyles across these three orders has accompanying differences in the cardiovascular anatomy and physiology allowing for adaptations to aquatic or terrestrial habitats, pulmonic or gill respiration, hibernation, and body elongation (in the caecilian). This article provides a review of amphibian cardiovascular anatomy and physiology with discussion of unique species adaptations. In addition, amphibians as cardiovascular animal models and commonly encountered natural diseases are covered. PMID- 19131030 TI - Normal reptile heart morphology and function. AB - Major differences among reptile taxa include the shape of the heart, degree of separation of the ventricular compartments, degree of development of the intraventricular muscular ridge, and in crocodilians, the interventricular septum. In many cases, the structural-functional features of the reptilian heart provide adaptive plasticity, allowing for the ecological and behavioral diversity seen. As a result, variation may surface in clinical measures of cardiac performance. This article updates clinical context, provides an understanding of the variation in reptilian cardiovascular systems, and their functional implications for the assessment and treatment of reptile patients. PMID- 19131031 TI - Reptile cardiology. AB - Cardiovascular disease in reptiles generally is considered an uncommon finding in captive animals, but no large-scale, cross-sectional studies have been performed to determine its prevalence. It is possible that cardiovascular disease is more common than is generally accepted and that the current belief results from limited clinical and diagnostic experience. This article offers guidance drawn from the author's clinical experience and the available literature. It is important that veterinarians pursue a thorough history, physical examination, and diagnostic work-up when managing cardiovascular disease in a reptile case. Veterinarians working with these cases should document their findings and share them with their colleagues to build an evidence-based foundation for reptile medicine. PMID- 19131032 TI - Cardiovascular physiology and diseases of pet birds. AB - Avian cardiac disease in pet birds occurs more often than previously assumed. The article focuses on anatomic peculiarities of the avian cardiovascular system and common diseases. Diagnostic possibilities are demonstrated, and therapeutic measures are discussed. PMID- 19131033 TI - Cardiovascular anatomy, physiology, and disease of rodents and small exotic mammals. AB - Cardiovascular disease in small exotic mammals is anecdotally common, but clinical reports of diagnosis and treatment of disease are rare. This article focuses on known causes of cardiovascular disease in the small exotic mammal. Normal anatomy and physiology, as it differs from the dog and cat, is also highlighted. Cardiomyopathy, dirofilariasis, atrial thrombosis, and other acquired and congenital cardiac and vascular diseases of rodents, hedgehogs, sugar gliders, raccoons, opossums, and skunks are reviewed. Expected clinical signs and diagnostic and treatment options, including a formulary, are provided for these species. PMID- 19131034 TI - Ferret cardiology. AB - Cardiac disease in pet ferrets is common and includes dilated cardiomyopathy, arrhythmias, and acquired valvular disease. Clinical presentation of cardiac disease in ferrets may be similar to dog or cats, although hind limb weakness may be a prominent feature. Radiography, ECG, and ultrasound are all useful tools in the diagnosis of cardiac disease in ferrets. Therapeutics for cardiac disease in ferrets is based on recommendations for dogs and cats. The prognosis for cardiac disease in ferrets varies from fair to guarded, depending on underlying disease. PMID- 19131035 TI - Cardiovascular physiology and diseases of the rabbit. AB - This article reviews what is known about the diagnosis and management of cardiovascular diseases in the pet rabbit. Current knowledge is based on anecdotal reports, derived from research data using the rabbit as an animal model of human cardiovascular diseases, but most importantly canine and feline cardiology. It is likely that, as cardiovascular diseases are more often recognized, more specific information will soon become available for the treatment of the pet rabbit with cardiac disease. PMID- 19131036 TI - Levodopa in the early treatment of Parkinson's disease. AB - l-dopa has many advantages as initial therapy for Parkinson's disease (PD). It is safer, more efficacious, associated with fewer adverse effects, few interactions, easier for patients to use and for clinicians to prescribe, and cheaper than dopamine (DA) agonists. Although l-dopa is more likely than DA agonists to introduce motor fluctuations and dyskinesia, l-dopa is also more effective in improving motor function. Furthermore, there is no long-term benefit from delaying l-dopa based on the risk of motor complications or psychiatric symptoms. Many investigations have shown that l-dopa does not accelerate disease progression. Now is the time to re-evaluate l-dopa for initial treatment of PD. PMID- 19131037 TI - Inter- and intra-individual variation in L-dopa pharmacokinetics in the treatment of Parkinson's disease. AB - Parkinson's disease is a neurodegenerative, slowly progressive, age-related disorder. Numerous medications have been developed for its treatment and the prognosis of the disorder has improved greatly over recent years. However, the effects of medicines are variable among patients, and there are also daily fluctuations in the effects of medications in the same person. The factors that cause individual variations in the effects of medicines, the causes, and strategies to cope with these fluctuations are reviewed. PMID- 19131038 TI - Reevaluation of levodopa therapy for the treatment of advanced Parkinson's disease. AB - Levodopa is the most appropriate drug in theory for supplementing dopamine deficiency in the brain of Parkinson's disease (PD) patients. In consideration of the pharmacological properties of levodopa, measurement of 3,4 dihydroxyphenylalanine (DOPA) plasma concentration is significant and important in daily medical care. Akinesia of advanced PD patients comprises a combination of two distinct symptoms, hypokinesia and bradykinesia. It is probable that hypokinesia in PD does not originate from failure of neural pathways from the substantia nigra to motor striatum but is associated with dysfunction of the limbic striatum. Herein the pathophysiologic condition of the limbic striatum in PD patients is discussed and reasons suggested why drug efficacy of dopamine replenishment in this system is meager. PMID- 19131039 TI - L-dopa therapy for Parkinson's disease: past, present, and future. AB - Dopamine (DA) supplementation therapy by l-dopa for Parkinson's disease (PD) was established around 1970. The dose of l-dopa can be reduced by the combined administration of inhibitors of peripheral l-amino acid decarboxylase (AADC), catechol O-methyltransferase (COMT), or monoamine oxidase B (MAO B). DA in the striatum may be produced from exogenously administered l-dopa by various AADC containing cells, such as serotonin neurons. The long-term administration of l dopa in PD patients may produce l-dopa-induced dyskinesia (LID), which may be due to chronic overstimulation of supersensitive DA D1 receptors. l-dopa may be used in combination with various new strategies such as gene therapy or transplantation in the future. PMID- 19131040 TI - Levodopa therapeutics for Parkinson's disease: new developments. AB - Levodopa serves as the gold standard of anti-parkinsonian therapy and nearly every patient with Parkinson's disease eventually receives this drug. To improve upon levodopa therapy, several forms of treatment have been devised to augment its actions, and new delivery systems are under development. This new research offers promise for improving outcomes with this highly effective therapy. PMID- 19131041 TI - Toxic effects of dopamine metabolism in Parkinson's disease. AB - Levodopa is the most effective medication for Parkinson's disease (PD). In contrast, there is evidence that levodopa and its metabolites such as dopa/dopamine quinone are toxic for nigral neurons based on in vitro studies. Moreover, there is growing evidence that oxidative stress and mitochondrial dysfunction contribute the pathogenesis of PD. Thus, studies for oxidative stress give us good information for elucidating the pathogenesis of PD. In this regard, it is mandatory to develop markers such as 4-hydroxy-nonenal (HNE). HNE is a product of lipid peroxidation. Indeed, immunohistochemical studies have revealed that HNE-modified proteins accumulate within ragged red fibers (RRFs). This finding indicated that mitochondrial impairment may be linked to oxidative stress. Moreover, HNE-modified proteins accumulate in nigral neurons. In PD, mitochondrial dysfunction such as complex I deficiency has also been reported. In addition, HNE can modify alpha-synuclein (SNCA). Subsequently, this modification may trigger the aggregation of this protein. At a minimum, this modification could be associated with oligomer formation or fibrillation of SNCA. PMID- 19131042 TI - Neuroprotective and toxic changes in microglia in neurodegenerative disease. AB - Microglia are macrophage-like cells in the CNS. As macrophages, activated microglia remove potentially deleterious debris and promote tissue repair. However, they can release potentially cytotoxic substances in vitro. So-called fully activated microglia, observed at the injury site in many neurodegenerative conditions, are neurotoxic. This suggests that some factor(s) may contribute to change microglial phenotype from protective to toxic, but details are not clear. Recently, we generated HIV-derived Nef protein-transduced microglia. They increase the potential to produce O(-)(2) and MPO-like peroxidase activity, resulting in neurotoxicity. Therefore, the target protein(s) of Nef might be involved in the control of microglial neurotoxicity. PMID- 19131043 TI - Parkin gene therapy. AB - The search for genetic mutations responsible for familial forms of Parkinson's disease (PD) has identified several genes and loci. Mutations in the parkin gene are linked to autosomal recessive juvenile parkinsonism. The genetic forms of PD are uncommon, but gene therapy targeting alpha-synuclein, parkin, or other pathways may be also applicable for idiopathic PD. Intriguingly, several studies suggest that parkin gene therapy could be useful in a subset of PD patients with mutations in the alpha-synuclein gene. Furthermore, if parkin overexpression can correct neuronal degeneration in the substantia nigra and striatum, it might be a potentially effective therapy for alpha-synucleinopathy. PMID- 19131044 TI - Emotional impairment in Parkinson's disease. AB - Patients with Parkinson's disease (PD) may show emotional impairment in the early stages of the disease. PD patients show disadvantageous decision-making, which is related to decreased emotional responses, as measured by skin conductance responses (SCRs). This pattern of decreasing SCRs is similar to that observed in amygdala-damaged patients. In facial expression recognition, PD patients did not show amygdala activation. In another study, PD patients did not show amygdala activations to unpleasant olfactory stimuli, which were observed in normal controls. Emotional impairment in PD patients may reflect amygdala dysfunction in early PD. PMID- 19131045 TI - Neuroimaging of L-dopamine and dopamine agonist therapy - revisited. AB - Treatment of Parkinson's disease (PD) is improving and we can choose a variety of drugs and surgical procedures for appropriate stages of the disease. However, we depend upon clinical signs and symptoms as well as patient reports to evaluate such issues. While neurochemical research into the etiological aspects of PD has been expanding, imaging is still under investigation. Here, I summarize the results of studies with regard to imaging of dopaminergic systems, and discuss the key points for imaging of PD with respect to treatment evaluation. PMID- 19131046 TI - The hypothesis of the genesis of motor complications and continuous dopaminergic stimulation in the treatment of Parkinson's disease. AB - The symptoms of Parkinson's disease can become increasingly difficult to control as the disease advances. Levodopa remains the most efficacious therapy for symptomatic treatment. However, with long-term therapy motor and non-motor complications develop. There is now accumulating evidence that several factors - the progressive pathology of Parkinson's disease, the change in drug pharmacodynamics, and the pulsatile manner in which short-acting dopaminergic agents stimulate striatal dopamine receptors - are key contributors to the priming of the basal ganglia for induction of motor complications. In this paper, an overview of the different factors inducing motor fluctuations and the role of dopamine receptors stimulation is provided. PMID- 19131047 TI - Effect of face form on maximal molar bite force with natural dentition. AB - OBJECTIVE: The purpose of this study was to determine mean maximum bite force in adults with normal occlusion and to evaluate effect of face form on it. DESIGN: Twenty male and 20 female students with normal dentitions and between the ages of 19 and 27 participated in the study. A strain-gauge force transducer was developed to measure bite force and was calibrated with known loads. Three measurements were performed on each side of the dentition in the first molar region and mean values used for analysis. Face form was defined as square, tapering, square-tapering or oval and determined using digital photographs. Effect of gender and face form on bite force was statistically analysed using two way ANOVA and Duncan tests. RESULTS: Mean maximum bite force and standard deviation (S.D.) in the sample population was 64.4 (24.0) kilograms (kg). In men, the mean was 73.6 (23.8) which was statistically higher than in women (53.0 (19.6) kg) (P < 0.05). Mean maximum bite force in subjects with square face form was 93.7 kg, which was significantly higher than in subjects with other facial forms (P < 0.05). CONCLUSION: The results showed higher bite force in men and those with square face form. Square face form may contribute to higher bite force values by maintaining a higher mechanical advantage for muscles of mastication. PMID- 19131048 TI - Isolation and characterization of poly- and oligosaccharides from the red microalga Porphyridium sp. AB - The current study forms part of an ongoing research effort focusing on the elucidation of the chemical structure of the sulfated extracellular polysaccharide of the red microalga Porphyridium sp. (UTEX 637). We report here on the chemical structure of a fraction separated from an acidic crude extract of the polysaccharide, as investigated by methylation analysis, carboxyl reduction methylation analysis, desulfation-methylation analysis, partial acid hydrolysis, Smith degradation, together with 1D and 2D (1)H and (13)C NMR spectroscopy. This fraction with a molar mass of 2.39x10(5)g mol(-1) comprised D- and L-Gal, D-Glc, D-Xyl, D-GlcA, and sulfate groups in a molar ratio of 1.0:1.1:2.1:0.2:0.7. The almost linear backbone of the fraction is composed of (1-->2)- or (1-->4)-linked d-xylopyranosyl, (1-->3)-linked L-galactopyranosyl, (1-->3)-linked D glucopyranosyl, and (1-->3)-linked D-glucopyranosyluronic acid and comprises a possible acidic building unit: [(2 or 4)-beta-D-Xylp-(L-->3)]m-alpha-D-Glcp-(1- >3)-alpha-D-GLCPA-(1-->3)-L-Galp(l-->. Attached to the backbone are sulfate groups and nonreducing terminal D-xylopyranosyl and galactopyranosyl residues, which occur at the O-6 positions of Glc-derived moieties in the main chain. PMID- 19131049 TI - A straightforward alpha-selective aromatic glycosylation and its application for stereospecific synthesis of 4-methylumbelliferyl alpha-T-antigen. AB - A practical and efficient alpha-selective aromatic glycosylation with simple per O-acetyl glycopyranosyl trichloroacetimidates is reported. The method is particularly effective for L-fucosyl and 2-azido-2-deoxy-D-galatosaminyl imidates, with which exclusive alpha-selectivity was achieved. The synthetic utility of this method was demonstrated in the stereoselective synthesis of 4 methylumbelliferyl alpha-T-antigen. PMID- 19131050 TI - The dynamic nature of knowledge: insights from a dynamic field model of children's novel noun generalization. AB - This paper examines the tie between knowledge and behavior in a noun generalization context. An experiment directly comparing noun generalizations of children at the same point in development in forced-choice and yes/no tasks reveals task-specific differences in the way children's knowledge of nominal categories is brought to bear in a moment. To understand the cognitive system that produced these differences, the real-time decision processes in these tasks were instantiated in a dynamic field model. The model captures both qualitative and quantitative differences in performance across tasks and reveals constraints on the nature of children's accumulated knowledge. Additional simulations of developmental change in the yes/no task between 2 and 4 years of age illustrate how changes in children's representations translate into developmental changes in behavior. Together, the empirical data and model demonstrate the dynamic nature of knowledge and are consistent with the perspective that knowledge cannot be separated from the task-specific processes that create behavior in the moment. PMID- 19131051 TI - Cognitive neuroscience of drawing: contributions of neuropsychological, experimental and neurofunctional studies. PMID- 19131052 TI - Shortness of breath at night and health status in congestive heart failure: effects of environmental conditions and health-related and dietary factors. AB - OBJECTIVES: Recent studies suggest that persons with congestive heart failure (CHF) may be at higher risk for the short-term effects of air pollution. We carried out this daily diary panel study in Montreal, Quebec, to determine whether indicators of self-reported health status and shortness of breath at night were associated with selected health-related and dietary factors, weather conditions, and air pollution. METHODS: Thirty-one subjects with CHF participated in this study in 2002 and 2003. Over a 2-month period, they measured their oxygen saturation, pulse rate, weight, and temperature each morning and recorded these and other data in a daily diary. Every morning they recorded on visual analogue scales their assessments of their general health, shortness of breath, and weakness, their weight, temperature and other data in a daily diary. Air pollution and weather conditions were obtained from fixed-site monitoring stations. This paper deals only with the daily self-reported health outcomes of general health and shortness of breath. We made use of mixed regression models, adjusting for within-subject serial correlation and temporal trends, to determine the association between oxygen saturation and pulse rate and health-related, dietary and environmental variables. Depending on the model, we accounted for the effects of a variety of health-related and dietary variables as well as NO(2), ozone, maximum temperature and change in barometric pressure at 8:00AM from the previous day. RESULTS: Of the many associations for self-reported general health and shortness of breath, we found only a few statistically significant predictors, although increases in many variables showed decrements in self reported general health and shortness of breath. The statistically significant associations with general health were increases in daily pulse rate and body weight, higher maximum ambient air temperature, higher relative humidity, and ozone (on the concurrent day). Statistically significant predictors of worsened shortness of breath at night were increases in blood pressure, body weight, and higher maximum ambient air temperature (lag 0 and 1 days). We also found that there was little confounding between environmental variables and health and dietary factors. CONCLUSIONS: The findings from the present investigation suggest that certain health-related indices and environmental conditions affect self reported health and shortness of breath in CHF patients, although larger studies are needed to confirm these findings. PMID- 19131053 TI - Assessment of airborne microorganism contamination in an industrial area characterized by an open composting facility and a wastewater treatment plant. AB - In order to assess the potential exposure hazard to workers and people living in the immediate surroundings of an area characterized by an open composting facility and a wastewater treatment plant, a quantitative and a qualitative analysis of airborne microorganisms were carried out. Air sampling was performed once a week for four consecutive weeks in summer and winter. Six sites were selected as air sampling sites: one was upwind at approximately 40 m from the facilities; the other five were downwind at increasing distances from the facilities, with the furthest at 100 m away. Monitoring permitted us to verify the influence that the composting activities and wastewater treatment had on the bacterial and fungal contamination of the air. The results obtained have been expressed by means of contamination indexes that have already been used in previous works: a major microbiological contamination near the plants was evidenced. Near the facilities, mesophilic bacteria, psychrophilic bacteria and microfungi showed the highest median concentrations, respectively, of 307.5, 327.5 and 257.5 CFU/m(3). Moreover, the season generally influenced the concentration of the bacteria as well as of the fungi; higher in summer than in winter. The contamination index global index of microbial contamination (GIMC/m(3)) showed mean values of 4058.9 in summer and 439.7 in winter and the contamination index-amplification index (AI) showed values of 4.5 and 1.1 in the same seasons, respectively. Controlling the seasonal effect, mesophilic bacteria, Pseudomonas spp. and Enterobacteriaceae showed a significant decline in concentration with respect to upwind air samples and with increasing distance. Both GIMC and AI showed a significant decline with respect to upwind air samples by increasing the distance from facilities after adjusting for the seasonal effect. In conclusion, even if these plants do not represent a potential risk for nearby populations, they may pose a potential health risk for workers. PMID- 19131054 TI - Modulation of HOXA10 and other markers of endometrial receptivity by age and human chorionic gonadotropin in an endometrial explant model. AB - OBJECTIVE: To characterize the endometrial response to human chorionic gonadotropin (hCG), as influenced by uterine age, using endometrial receptivity markers including HOXA10, vascular endothelial growth factor (VEGF), and glycodelin in an endometrial explant culture system. DESIGN: In vitro molecular biology research. SETTING: Academic infertility clinic and molecular biology laboratory. PATIENT(S): Fourteen prospective recipients of egg donation (mean age, 44 +/- 8 years). INTERVENTION(S): Subjects received cyclical estrogen (E(2)) and progesterone (P(4)) and underwent an endometrial biopsy on day 7 of P(4). Endometrial biopsy samples were cut into 1-mm(3) pieces and cultured in Dulbecco's modified Eagle's medium/Ham's F-12 with E(2) and P(4), without (control) or with hCG (400, 4000, and 40,000 mIU/mL) on Millicell-CM inserts for 24 hours. MAIN OUTCOME MEASURE(S): Explant viability was assessed using immunohistochemistry (IHC). Semiquantitative polymerase chain reaction was performed to evaluate relative gene expression via mRNA levels of HOXA10, VEGF, and glycodelin. RESULT(S): Explant viability was confirmed on IHC by histology and Ki-67 staining, a marker of proliferation. HOXA10, VEGF, and glycodelin gene expression increased at all concentrations of hCG over those of controls. HOXA10 gene expression was inversely correlated with age (-0.08- +/- 0.03-fold decrease in gene expression/year of age). CONCLUSION(S): The endometrial explant culture system is a promising model for the study of endometrial response as it maintains interactions among the stroma, glands, and epithelium. HOXA10, VEGF, and glycodelin all demonstrated increased gene expression in response to increasing hCG concentrations, supporting the role of hCG as a candidate protein for blastocyst-endometrial communication. Statistically significant associations between age and expression of HOXA10 provide novel evidence that uterine age may play a role in endometrial response on a molecular level. PMID- 19131055 TI - A comparison of gonadotropin-releasing hormone (GnRH) antagonist and GnRH agonist flare protocols for poor responders undergoing in vitro fertilization. AB - OBJECTIVE: To compare stimulation profiles, pregnancy, and live birth rates in poor responders during in vitro fertilization (IVF) cycles using either a gonadotropin-releasing hormone (GnRH) antagonist (cetrorelix) or a GnRH agonist flare protocol (leuprolide). DESIGN: Retrospective chart review. SETTING: A university-affiliated IVF program. PATIENT(S): Women designated as poor responders based on a prior stimulation cycle or baseline follicle-stimulating hormone (FSH) level of >10 mIU/mL, who needed at least 375 IU of starting daily gonadotropins in the study cycle. INTERVENTION(S): Administration of GnRH agonist flare or GnRH antagonist protocol. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate, live birth rate. RESULT(S): For 68 GnRH antagonist and 45 GnRH agonist flare cycles, the groups were similar with respect to age (38.8 versus 38.6 years) and basal FSH concentration (8.33 versus 8.65 mIU/mL). No statistically significant differences between the protocol types were noted in peak estradiol levels, amount of gonadotropins used, number of oocytes obtained, or embryos transferred. The pregnancy rates (40% versus 45.2%) and live birth rates (27.7% versus 31.7%) in the GnRH antagonist and flare groups, respectively, were similar. CONCLUSION(S): We achieved excellent and comparable pregnancy and live birth rates in poor responders of advanced reproductive age with the use of either GnRH antagonist or flare protocol. PMID- 19131056 TI - Magnetic resonance imaging is useful for diagnosis and evaluation of recurrent intravenous leiomyomatosis before surgery. AB - A case of recurrent intravenous leiomyomatosis intruding into the inferior vena cava is presented. Magnetic resonance imaging provided important information for diagnosis and evaluation of the disease before surgery. PMID- 19131057 TI - Association study of vascular endothelial growth factor polymorphisms with the risk of recurrent spontaneous abortion. AB - OBJECTIVE: To investigate the association of vascular endothelial growth factor (VEGF) polymorphisms (-2578C>A, -1154G>A, -634G>C, 936C>T) with idiopathic recurrent spontaneous abortion (RSA) in Koreans. DESIGN: Prospective case-control study. SETTING: University-based hospital. PATIENT(S): Two hundred and fifteen patients with a history of two or more unexplained consecutive pregnancy losses and 113 healthy controls with at least one live birth and no history of pregnancy loss. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses were performed for the -2578C>A and 936C>T genotypes. Real-time PCR was also used to analyze the -1154G>A and -634G>C genotypes. RESULT(S): The GA (adjusted odds ratio [AOR] 2.774; 95% confidence interval [CI] 1.512-5.092) genotype of the VEGF -1154G>A polymorphism was significantly different between women with idiopathic RSA and controls. The difference in overall (GA + AA) frequency was also marginally significant between the controls and patients with idiopathic RSA (AOR, 2.006; 95% CI, 1.158-3.473). The differences in frequencies of the A-A-G-T and C-A-G-T haplotypes of the VEGF polymorphisms (-2578C>A, -1154G>A, -634G>C, 936C>T) were marginally significant between the patient and control groups. CONCLUSION(S): This study suggests that VEGF polymorphisms and haplotypes are a genetic determinant for the risk of idiopathic RSA in Korean women. PMID- 19131058 TI - Mock embryo transfer does not affect uterine contractility. AB - OBJECTIVE: To assess the degree of utero-stimulation induced by transcervical introduction of a catheter as in mock ET. DESIGN: Prospective study. SETTING: Reproductive medicine unit. PATIENT(S): Eighty volunteers undergoing mild ovarian stimulation and IUI. INTERVENTION(S): Just before IUI, women had uterine contraction (UC) frequency assessed with 2-minute sagittal ultrasound scans of the uterus (precatheter measurement). Thereafter, one of three types of catheters, each with a different stiffness level (low, n = 25; medium, n = 30; or high, n = 25) was introduced into the cervix up to the inner cervical os, then immediately withdrawn, and the UC frequency reassessed (postcatheter measurement). MAIN OUTCOME MEASURE(S): Percentage change in UC frequency. RESULT(S): Median (range) UC frequency was not significantly different at pre- and postcatheter measurements: 2.0 (0.0-4.5) UC per minute and 2.2 (0.0-5.5) UC per minute. In addition, the stiffness of the catheter (low, medium, or high) did not influence the UC frequency, with median (range) percentage of change from pre to postcatheter measurement at 0 (-100% to +75%), 0 (-100% to +100%), and 0 ( 40% to +100%), respectively. CONCLUSION(S): The introduction of a catheter up to the inner cervical os, as usually performed in mock ET, does not stimulate uterine contraction frequency, irrespective of catheter stiffness. PMID- 19131059 TI - Plasma interleukin-18 levels are increased in the polycystic ovary syndrome: relationship of carotid intima-media wall thickness and cardiovascular risk factors. AB - OBJECTIVE: To determine serum interleukin (IL)-18 levels and to find out whether IL-18 is associated with carotid intima-media wall thickness (IMT) and various cardiovascular risk factors in women with polycystic ovary syndrome (PCOS). DESIGN: A prospective, controlled study. SETTING: University hospital. PATIENT(S): Sixty women with PCOS and 60 healthy women were included this study. INTERVENTION(S): Serum levels IL-18, homocysteine (Hcy), C-reactive protein (CRP), IL-6, malonyldialdehyde (MDA), lipid and hormone profiles were measured. Carotid IMT was evaluated for both common carotid arteries. MAIN OUTCOME MEASURE(S): Serum IL-18, carotid IMT, Hcy, CRP, IL-6, MDA, and homeostasis model assessment of insulin resistance. RESULT(S): The evaluation, which was made without the obesity influence taken into consideration, revealed that patients with PCOS have increased serum IL-18 levels than that of the control group (214 +/- 102 vs. 170 +/- 78 pg/mL). The interaction between PCOS and obesity was seen to have statistical significance (F = 67.8). Body mass index (BMI), waist to-hip ratio, Hcy, and homeostasis model assessment of insulin resistance are independent determinants of plasma IL-18 in patients with PCOS. Elevated serum IL 18 levels were positively and significantly correlated with a greater carotid IMT. For Hcy and carotid IMT, the interaction between PCOS and obesity was found in a two-way ANOVA variation analysis (F = 48.5 and F = 81.5, respectively). CONCLUSION(S): Elevated serum IL-18 levels were associated with cardiovascular risk factors and carotid IMT in patients with PCOS. PMID- 19131060 TI - Peritoneal ectopic lesions from women with endometriosis show abnormalities in progesterone-dependent glycan expression. AB - Examination of 12 paired peritoneal ectopic and eutopic endometria for histochemical binding of Dolichos biflorus agglutinin, normally found in the mid late secretory part of the cycle, showed a failure of lectin binding in 9 of 11 secretory-phase lesions although the eutopic specimens generally stained normally. This failure of glycan expression in the secretory phase may result from various anomalies, including an inability to respond to progesterone, possibly due to a lack of, or to nonfunctional, progesterone receptors, suggesting that an ectopic environment may produce changes in tissue cell biology and hormonal responsiveness compared with that of eutopic endometrium. PMID- 19131062 TI - Increasing incidence of injuries and fatalities inflicted by wild animals in Kashmir. AB - OBJECTIVE: To clarify the causes of the increased incidence of injuries inflicted by wild animals in Kashmir, and to suggest preventive measures. METHODS: A retrospective study reviewed records of these injuries. Data were collected from the hospital in Srinagar and from the Wildlife Protection Department of Kashmir. RESULTS: A total of 203 attacks (26 deaths and 177 near-fatal injuries) were recorded from January 2005 to October 2007, involving 145 (71.5%) male and 58 (28.5%) female victims. The attacking animal was a black bear in 104 (51.2%), a leopard in 18 (8.8%), a wolf in 7 (3.4%) and unidentified in 74 (36.4%) cases; 130 (64%) of these attacks occurred in southern Kashmir. CONCLUSION: The steady increase in human population in areas close to jungle results in deforestation and destruction of wildlife habitat. The reduction in monkeys and deer (the staple food of leopards) causes these dangerous predators to search for food in the areas occupied by humans, who may then be attacked. Treatment of these cases requires a multidisciplinary approach, including an orthopaedic surgeon, plastic surgeon, microbiologist and psychiatrist, to achieve best cosmetic and functional results. PMID- 19131061 TI - Clinical predictors of occult pneumothoraces in severely injured blunt polytrauma patients: A prospective observational study. AB - INTRODUCTION: The supine antero-posterior (AP) chest radiograph (CXR) is an insensitive test for detecting post-traumatic pneumothoraces (PTXs). Computed tomography (CT) often identifies occult pneumothoraces (OPTXs) that were not diagnosed on CXR. The purpose of this study was to prospectively determine the incidence, and validate previously identified clinical predictors, of OPTXs after blunt trauma. METHODS: All severe blunt injured patients (injury severity score (ISS)>or=12) presenting to a level 1 trauma centre over a 17-month period were prospectively evaluated. Thoracoabdominal CT scans and corresponding CXRs were reviewed at the time of admission. Patients with OPTXs were compared to those with overt PTXs regarding incidence and previously identified predictive risk factors (subcutaneous emphysema, rib fractures, female sex and pulmonary contusion). RESULTS: CT imaging was performed concurrent to CXR in 405 blunt trauma patients (ISS>or=12) during the study period. PTXs were identified in 107 (26%) of the 405 patients. Eighty-one (76%) of these were occult when CXRs were interpreted by the trauma team. Concurrent chest trauma predictive of OPTXs was limited to subcutaneous emphysema (p=0.003). Rib fractures, pulmonary contusions and female sex were not predictive. CONCLUSIONS: OPTXs were missed in up to 76% of all seriously injured patients when CXRs were interpreted by the trauma team. This is higher than previously reported in retrospective studies and is likely based on the difficult conditions in which the trauma team functions. Subcutaneous emphysema remains a strong clinical predictor for concurrent OPTXs. PMID- 19131063 TI - No association of two functional polymorphisms in human ALOX15 with myocardial infarction. AB - The 12/15-lipoxygenase plays a janus-role in inflammation with pro-inflammatory and anti-inflammatory effects in cell systems and even opposite effects on atherosclerosis in two different animal species. Screening of the human 15 lipoxygenase (ALOX15) gene detected a polymorphic C to T substitution at position c.-292, which led to three times higher ALOX15 activity in macrophages and showed a trend to be atheroprotective in a small case-control study for coronary artery disease (CAD). A second polymorphism at position c.1693C>T leading to an T560M exchange and an inactive enzyme was recently associated with increased CAD. We now investigated whether these polymorphisms or a certain haplotype of ALOX15 are associated with myocardial infarction (MI) in a case-control subset from the population-based MONIKA/KORA cohort S3. Six polymorphisms in ALOX15 were analyzed in 2629 participants to cover all major haplotypes with a frequency higher than 1% in the Caucasian population. None of the polymorphism was associated with MI but a rare ALOX15 haplotype showed a significant protective effect on the risk for MI (p=0.03). However, none of the polymorphisms or haplotypes was associated with CRP levels. These data suggest that ALOX15 may play a less prominent role during later stages of atherosclerosis involving atherothrombotic mechanisms than eventually during early plaque development. PMID- 19131064 TI - Interleukin 17: an unlikely marker of acute coronary syndrome? PMID- 19131065 TI - Effects of extended-release niacin on lipid profile and adipocyte biology in patients with impaired glucose tolerance. AB - BACKGROUND: Low high-density lipoprotein cholesterol (HDL-C) serum concentrations are independent risk factors for the development of coronary artery disease. In patients with the metabolic syndrome, low HDL-C can contribute to premature atherosclerosis. Extended-release (ER) niacin increases HDL-C and was shown to slow the progression of atherosclerosis. Adipose tissue is an important site of niacin action. Here we sought to determine potential pleiotropic effects of ER niacin on adipose tissue biology in patients with impaired glucose tolerance (IGT). METHODS AND RESULTS: Thirty patients with IGT (mean age=45.2+/-3.9 years), low HDL-C serum concentrations (HDL-C <1.0 mmol/l), but no additional comorbidities were treated once-daily with ER niacin (1000 mg) in a randomized open-label controlled (n=30) study for 6 months. During the first 4 weeks, daily dose was increased from 375 to 1000 mg in weekly intervals. At baseline and after 6 months, subcutaneous adipose tissue biopsies were taken, body fat mass, insulin sensitivity (euglycemic-hyperinsulinemic clamp), and adipokine serum concentrations were measured. After 6 months of ER niacin treatment, HDL-C increased significantly by 24% and adiponectin by 35%. In addition, ER niacin significantly reduced circulating lipoprotein (a) by 38% (p<0.001) and fasting triglycerides by 12% (p<0.05). Whole-body insulin sensitivity increased in the ER niacin treatment group, although this trend was not statistically significant (p=0.085). Six months ER niacin led to a significant reduction in mean adipocyte size associated with increased insulin sensitivity in isolated adipocytes and gene expression changes including increased adiponectin, C/EBPalpha, C/EBPdelta, PPARgamma and decreased carnitine palmitoyl transferase 2, hormone sensitive lipase, nicotinic acid receptor (GPR109B) and fatty-acid synthase mRNA expression. CONCLUSION: Treatment with ER niacin significantly improves atherogenic lipid profile in patients with IGT. These beneficial effects could at least in part be due to pleiotropic niacin effects in adipose tissue, characterized by decreased mean adipocyte size, increased insulin sensitivity and altered mRNA expression profile. PMID- 19131066 TI - Destabilizing and stabilizing forces to assess equilibrium during everyday activities. AB - Postural stability is essential to functional activities. This paper presents a new model of dynamic stability which takes into account both the equilibrium associated with the body position over the base of support (destabilizing force) and the effort the subject needs to produce to keep his/her centre of mass inside the base of support (stabilizing force). The ratio between these two forces (destabilizing over stabilizing) is calculated to provide an overall index of stability for an individual. Preliminary results from data collected during walking at preferred and maximal safe speed in four older adults (aged from 64 to 84yr) showed that both forces are lower for subjects with reduced maximal gait speed. In addition, the stabilizing force increases by 2-3 times from preferred to maximal speed, while the destabilizing force barely changes with gait speed. Overall, the model through the index of stability attributes lower dynamic stability to subjects with lower maximal gait speed. These preliminary results call for larger-scale studies to pursue the development and validation of the model and its application to different functional tasks. PMID- 19131067 TI - Forensic differentiation of biogenic organic compounds from petroleum hydrocarbons in biogenic and petrogenic compounds cross-contaminated soils and sediments. AB - "Total petroleum hydrocarbons" (TPHs) or "petroleum hydrocarbons" (PHCs) are one of the most widespread soil pollutants in Canada, North America, and worldwide. Clean-up of PHC-contaminated soils and sediments costs the Canadian economy hundreds of million of dollars annually. Much of this activity is driven by the need to meet regulated levels of PHC in soil. These PHC values are legally required to be assessed using standard methods. The method most commonly used in Canada, specified by the Canadian Council of Ministers of the Environment (CCME), measures the total hydrocarbon concentrations in a soil by carbon range (Fraction 1: C(6)-C(10); Fraction 2: C(10)-C(16), Fraction 3: C(16)-C(34): and Fraction 4: C(34)+). Using the CCME method, all of the materials extractible by a mixture of 1:1 hexane:acetone are considered to be petroleum hydrocarbon contaminants. Many hydrocarbon compounds and other extractible materials in soil, however, may originate from non-petroleum sources. Biogenic organic compounds (BOCs) is a general term used to describe a mixture of organic compounds, including alkanes, sterols and sterones, fatty acids and fatty alcohols, and waxes and wax esters, biosynthesized by living organisms. BOCs are also produced during the early stages of diagenesis in recent aquatic sediments. BOC sources could include vascular plants, algae, bacteria and animals. Plants and algae produce BOCs as protective wax coating that are released back into the sediment at the end of their life cycle. BOCs are natural components of thriving plant communities. Many solvent-extraction methods for assessing soil hydrocarbons, however, such as the CCME method, do not differentiate PHCs from BOCs. The naturally occurring organics present in soils and wet sediments can be easily misidentified and quantified as regulated PHCs during analysis using such methods. In some cases, biogenic interferences can exceed regulatory levels, resulting in remediation of petroleum impacts that are not actually present. Consequently, reliance on these methods can trigger unnecessary and costly remediation, while also wasting valuable landfill space. Therefore, it is critically important to develop new protocols to characterize and differentiate PHCs and BOCs in contaminated sediments. In this study, a new reliable gas chromatography-mass spectrometry (GC MS) method, in combination with a derivatization technique, for characterization of various biogenic compounds (including biogenic alkanes, sterols, fatty acids and fatty alcohols) and PHCs in the same sample has been developed. A multi criteria approach has been developed to positively identify the presence of biogenic compounds in soil and sediment samples. More than thirty sediment samples were collected from city stormwater management (SWM) ponds and wetlands across Canada. In these wet sediment samples, abundant biogenic n-alkanes, thirteen biogenic sterols, nineteen fatty carboxylic acids, and fourteen fatty alcohols in a wide carbon range have been positively identified. Both PHCs and BOCs in these samples were quantitatively determined. The quantitation data will be used for assessment of the contamination sites and toxicity risks associated with the CCME Fraction 3 hydrocarbons. PMID- 19131068 TI - Separation, detection and quantitation of peptides by liquid chromatography and capillary electrochromatography. AB - This review discusses different liquid chromatographic and capillary electrochromatographic approaches to the separation and quantitation of peptides using silica-based and polymeric-based columns with emphasis on liquid chromatography. Mass spectrometry detection and quantitation of peptides using labeled and label-free procedures, will also be discussed, as well as the effect of amino acids' properties on the solubility of peptides, an important parameter that influences the selection of the mobile phase. A discussion of different column packing materials, reversed-phase, cyclodextrins, macrocyclic antibiotics, porous graphitic carbon, mixed-phases, and normal-phase will be included, as well as a short discussion of multi-dimensional approaches for the separation of complex peptide mixtures. PMID- 19131069 TI - Retention characteristics of organic compounds on molten salt and ionic liquid based gas chromatography stationary phases. AB - The interest of using ionic liquids (ILs) as stationary phases in gas chromatography (GC) has increased in recent years. This is largely due to the fact that new classes of ILs are being developed that are capable of satisfying many of the requirements of GC stationary phases. This review highlights the major requirements of GC stationary phases and describes how molten salts/ILs can be designed to largely meet these needs. The retention characteristics of organic solutes will be discussed for ammonium, pyridinium, and phosphonium-based molten salts followed by imidazolium, pyridinium, pyrollidinium, and phosphonium-based IL stationary phases. The versatility of ILs allows for the development of stationary phases based on dicationic ILs, polymeric ILs, and IL mixtures. To aid in choosing the appropriate IL stationary phase for a particular separation, the reader is guided through the different types of stationary phases available to identify those capable of providing the desired separation selectivity of organic solutes while allowing for flexibility in ranges of temperature used throughout the separation. PMID- 19131070 TI - Sucralose screening in European surface waters using a solid-phase extraction liquid chromatography-triple quadrupole mass spectrometry method. AB - An analytical method was developed for the analysis of sucralose, a persistent chlorinated calorie-free sugar substitute, in surface waters. The method is based on solid-phase extraction (SPE) of 400mL water using Oasis HLB (Waters) adsorber material, followed by negative electrospray ionization (ESI) triple quadrupole LC MS-MS detection. Quantification was performed by external calibration, as well as by isotope dilution with deuterated sucralose d6 internal standard. Extraction with Oasis HLB, a polymeric adsorbent suited for polar compounds, was much more efficient at neutral pH than at pH 3; a recovery of 62+/-9% (n=6; determined at 1microg/L) could be achieved. Strong ion suppression caused by matrix substances was observed for sucralose in the SPE extracts. The analysis of 120 river surface water samples from 27 European countries showed that sucralose, which is in use in Europe since beginning 2005, can be found in the aquatic environment, at concentrations up to 1microg/L. Sucralose was predominately found in samples from the UK, Belgium, the Netherlands, France, Switzerland, Spain, Italy, Norway, and Sweden, suggesting an increased use of the substance in Western Europe. PMID- 19131071 TI - New gas chromatographic method for the enantioseparation of beta-amino acids by a rapid double derivatization technique. AB - A novel gas chromatographic method was developed for the enantioseparation of valuable acyclic and carbocyclic cis- and trans-beta-amino acids, including cispentacin and a number of its analogues and homologues. Excellent (in most cases baseline) separation was achieved for the racemates of these beta-amino acids on CP-Chirasil-Dex CB or CP-Chirasil L-Val columns after a simple and rapid double derivatization (esterification followed by N-acylation). The elution sequences were determined in all cases. PMID- 19131072 TI - Modeling the dynamic folding and surface-activity of a helical peptide adsorbing to a pendant bubble interface. AB - We have designed a peptide with switchable surface activity, where the folded (alpha-helical) form of the peptide is amphiphilic and the unfolded form is not. To understand the factors influencing the dynamics of the switchability, a model is developed for the transport of the surface active form of the peptide from the solution onto air-water interface. As is the case with the low molecular weight head-tail surfactants, the transport involves the bulk diffusion of the folded form to the surface and the kinetic adsorption onto the interface. Unlike the head-tail surfactants, the diffusion can be augmented by the kinetics of the folding of the peptide from the unfolded form. The model is formulated within the context of the transport of the peptide from a uniform bulk solution onto an initially clean air-water interface in a pendant bubble system, where the transport rate can be measured by recording the reduction in surface tension using the shape analysis of the bubble. Experiments are undertaken and compared to the predictions of the model simulations of the tension reduction for a range of values of the kinetic adsorption constant and the folding kinetic constant. The results indicate that the kinetic adsorption rate of the folded peptide onto air-water interface dominates the dynamic process, which contrasts many head-tail surfactants where diffusion typically dominates over kinetics adsorption. Moreover, our 'best-fits' suggest that there is a phase transition at high surface concentrations that slows the long-time adsorption of the peptides to the interface. Finally, the numerical solution is compared with an asymptotic solution, showing agreement with our findings that the fundamental dynamics of the tunable surface-active peptide are indeed controlled by the adsorption step. PMID- 19131074 TI - Shear modulus of colloidal suspensions: Comparing experiments with theory. AB - In this work, the experimentally determined shear modulus of a colloidal suspension has been compared to a calculated shear modulus based on an ordered lattice model. The experiments were performed on a well characterized calcite suspension. The calcite suspension was confirmed to be in a flocculated state, thus exhibiting strong elastic character. The experimental shear modulus was estimated from the elastic modulus in the linear viscoelastic region. This is contrary to established convention, where the high frequency limiting modulus is used. However, the network structure breaks down at high frequencies, resulting in a different system. Therefore the shear modulus was estimated from the response of the intact particle network structure, which resides within the linear viscoelastic region. Calculation of the shear modulus is based on an ordered lattice model, where the particles are arranged in regular 3D-arrays. Interaction forces between the particles are also considered, based on the well known DLVO-theory. Calculations of the shear modulus for flocculated colloidal suspensions are not trivial however, because of the random distribution of particles. Due to this fact the average interparticle distance is impossible to determine exactly and therefore the comparison between experimental and calculated shear modulus was made by calculating the shear modulus as a function of interparticle separation. Some different models for interaction forces between particles were tried, and the results were compared to rheological data. These preliminary results show that useful suspension properties can be evaluated by combining rheological measurements with theories for interactions between particles arranged in ordered lattices. PMID- 19131073 TI - The interaction between casein micelles and gold nanoparticles. AB - For the first time, the interaction of casein micelle (CM) with gold nanoparticles (GNPs) was studied using TEM, UV-visible, FTIR and fluorescence spectroscopy methods. The results show that GNPs bind to CM surface, leading to the formation of GNP-CM conjugates. GNPs exhibit similar particle diameters, and the CM structure is retained in GNP-CM conjugates. Furthermore, GNP-CM conjugates display good stability against salt concentration and pH. The combination of these measurements suggests that GNPs presumably bind to CM surfaces via complexation with the carboxylate or amine groups on CM surfaces, but not by electrostatic action. PMID- 19131075 TI - Study on modification of colloidal silica surface with magnesium ions. AB - The prepared colloidal silica via ion exchange was modified by adding Mg(+2) ions in the surface growth process of colloidal silica. The particle size of the modified colloidal silica was decreased by increasing the seed concentration, whereas that of colloidal silica was slightly increased as the seeds concentration exceed 15 wt%. But the trend of changes in surface area was completely reversed. Magnesium ions on surface of the modified colloidal silica were identified by the measurement of energy-dispersive X-ray spectrometry (EDS). After modifying with Mg(+2) ions, the iso-electric point (IEP) of colloidal silica in pH approximately 2 without modification, disappears over the whole pH range. PMID- 19131076 TI - Interlayer spacing effect of alkylammonium-modified montmorillonite on conducting and mechanical behaviors of polymer composite electrolytes. AB - Polymer composite electrolytes (PCE) based on poly(ethylene oxide) (PEO) and alkylammonium-functionalized clay materials were fabricated. Structural modification and the electrochemical properties were investigated in order to understand the effects of organically modified montmorillonite (OMMT) in the polymer matrix on the ion conductivity. Nanosize layer-structured clay fillers were used having an organic modifier with different alkyl lengths and positions. The X-ray diffraction (XRD) results revealed that the interlayer spacing (2.55 nm) for MMT-20A was increased compared with that (1.16 nm) for Na-MMT. Both the XRD and the thermal analysis results indicated that the PCE showed reduced crystallinity after the introduction of the OMMT fillers. From the ion conductivity results, the PCE containing MMT-20A showed higher conductivity (6.1x10(-4) S/cm) than that containing Na-MMT (2.2x10(-4) S/cm). This indicated that the improved ion conductivity was dependent on the reduced crystallinity that was correlated with the d-spacing of the MMT. Furthermore, the PCE/OMMT showed improved tensile strength. Finally, it was shown that the conducting and mechanical properties were dependent on the interlayer spacing of the modified clays. PMID- 19131078 TI - Guidelines for seagrass restoration: importance of habitat selection and donor population, spreading of risks, and ecosystem engineering effects. AB - Large-scale losses of seagrass beds have been reported for decades and lead to numerous restoration programs. From worldwide scientific literature and 20 years of seagrass restoration research in the Wadden Sea, we review and evaluate the traditional guidelines and propose new guidelines for seagrass restoration. Habitat and donor selection are crucial: large differences in survival were found among habitats and among donor populations. The need to preferably transplant in historically confirmed seagrass habitats, and to collect donor material from comparable habitats, were underlined by our results. The importance of sufficient genetic variation of donor material and prevention of genetic isolation by distance was reviewed. The spreading of risks among transplantation sites, which differed in habitat characteristics (or among replicate sites), was positively evaluated. The importance of ecosystem engineering was shown in two ways: seagrass self-facilitation and facilitation by shellfish reefs. Seagrass self facilitative properties may require a large transplantation scale or additional measures. PMID- 19131079 TI - Urging cautious policy applications of captive research data is not the same as rejecting those data. PMID- 19131080 TI - TBT-pollution in the Gulf of Thailand: a re-inspection of imposex incidence after 10 years. AB - Imposex in neogastropods was used to determine the relative TBT distribution in the Gulf of Thailand in 2006. To identify the imposex prevalence, 8757 specimens, belonging to 22 species from five families of neogastropods, were collected at 56 sites in 13 sample areas. These areas were located between the Bight of Bangkok in the north and the Malaysian border in the south. A contamination model was developed to compensate for differences in TBT sensitivity among species and to make comparisons among areas. At every area imposex was found in neogastropods. The highest incidence was in the east side of the Bight of Bangkok off Si Racha and Pattaya and in the southern part around Pattani. The same areas showed the highest frequency of imposex in 1996. While the frequency of imposex appeared to have slightly decreased in these areas, increases were found elsewhere. In Pattani Province this could be related to dumping of highly polluted harbour sediments in the shallow coastal area. The overall frequency of imposex in the Gulf of Thailand significantly increased from 1996 to 2006. PMID- 19131081 TI - Physcomitrella patens has lipoxygenases for both eicosanoid and octadecanoid pathways. AB - Mosses have substantial amounts of long chain C20 polyunsaturated fatty acids, such as arachidonic and eicosapentaenoic acid, in addition to the shorter chain C18 alpha-linolenic and linoleic acids, which are typical substrates of lipoxygenases in flowering plants. To identify the fatty acid substrates used by moss lipoxygenases, eight lipoxygenase genes from Physcomitrella patens were heterologously expressed in Escherichia coli, and then analyzed for lipoxygenase activity using linoleic, alpha-linolenic and arachidonic acids as substrates. Among the eight moss lipoxygenases, only seven were found to be enzymatically active in vitro, two of which selectively used arachidonic acid as the substrate, while the other five preferred alpha-linolenic acid. Based on enzyme assays using a Clark-type oxygen electrode, all of the active lipoxygenases had an optimum pH at 7.0, except for one with highest activity at pH 5.0. HPLC analyses indicated that the two arachidonic acid lipoxygenases form (12S)-hydroperoxy eicosatetraenoic acid as the main product, while the other five lipoxygenases produce mainly (13S)-hydroperoxy octadecatrienoic acid from alpha-linolenic acid. These results suggest that mosses may have both C20 and C18 based oxylipin pathways. PMID- 19131082 TI - Ultrastructural features and elemental distribution in eggshell during pre and post hatching periods in the green turtle, Chelonia mydas at Ras Al-Hadd, Oman. AB - Eggshells were randomly collected from turtle nests immediately after oviposition and at the end of incubation to examine the ultrastructural features using scanning JSM-5600LV microscopy. Three layers were recognized; an outer calcareous, a middle multistrata and an inner membrane. The calcareous layer had loose nodular units varying in shape and size without interlocking attachments. In freshly laid eggs, each nodular unit had spicules arranged in folded stacks. The spicules became unfolded during incubation, to form radiating configurations. Elemental composition and mapping of the layers were analyzed using energy dispersive spectroscopy (EDS). The elements were unevenly distributed throughout the eggshell and Ca(2+) decreased significantly after hatching. X-ray diffraction was used to identify the crystals of the eggshells. It revealed that nodular units of the calcareous were made up of CaCO(3), as aragonite (91%), calcite (6%) and vaterite (3%). The middle layer consisted of organic amorphous material with aragonite (89%) and calcite (11%). The shell membrane consisted of reticular fibers with crystals predominantly of NaCl halite. Thermogravimetry analysis of the calcareous layer indicated a complete evaporation of bonded H(2)O at 480 degrees C and CO(2) at 830 degrees C. Using the differential thermal analysis (DTA), aragonite was transformed to stable calcite at 425 degrees C. PMID- 19131083 TI - Characterization of novel VP7, VP4, and VP6 genotypes of a previously untypeable group A rotavirus. AB - Rotavirus is the most common cause of acute gastroenteritis among infants and young children throughout the world, but rotavirus cases in developing countries account for nearly all of the approximately 600,000 annual deaths. We studied the epidemiology of rotavirus in 22 rural communities in northern coastal Ecuador over a five-year period. From 250 rotavirus positive stool specimens, the percentage that could not be RT-PCR genotyped for VP4 and VP7 was 77% and 63%, respectively. The possibility of sample degradation was considered but discounted after an experimental examination of rotavirus stability and EM visualization of rotavirus-like particles in several untypeable samples. Finally, alternate primers were used to amplify Ecu534, a sample that was untypeable using most published VP4 and VP7 primers. Characterization of the VP7, VP4, and VP6 full gene segments revealed novel genotypes and nucleotide mismatches with most published primer sequences. When considered with other findings, our results suggest that primer mismatch may be a widespread cause of genotyping failure, and might be particularly problematic in countries with greater rotavirus diversity. The novel sequences described in this study have been given GenBank accession numbers EU805775 (VP7), EU805773 (VP4), EU805774 (VP6) and the RCWG has assigned them novel genotypes G20P[28]I13, respectively. PMID- 19131085 TI - Detection of pathogen based on the catalytic growth of gold nanocrystals. AB - A homogenous detection of pathogen (Giardia lamblia cysts) based on the catalytic growth of gold nanoparticles (AuNPs) has been studied. In this study, centrifugal filters were employed as tools to concentrate and separate the pathogen cells, and moreover amplify the detection signal. The catalytic growth of gold nanoparticles was verified to be positively related to gold seeds concentration. On this basis, homogenous detection of the pathogenic bacteria in liquid phase was established by means of conjugating antibody to gold seeds. Under the given experimental condition, detection limit of G. lamblia cysts was determined as low as 1.088 x 10(3) cells ml(-1). The additional nonspecific binding tests were also conducted to verify the detection specificity. This sensing platform has been proved to be a sensitive, reliable and simple method for large-scale pathogen detection, and provide valuable insight for the development of gold nanocrystals based colorimetric biosensors. PMID- 19131086 TI - A Fenton-like degradation mechanism for 1,4-dioxane using zero-valent iron (Fe0) and UV light. AB - In this study, the degradation mechanism of 1,4-dioxane using zero-valent iron (Fe0) in the presence of UV light was investigated kinetically. The degradation of 1,4-dioxane in Fe0-only, photolysis, and combined Fe0 and UV reactions followed the kinetics of a pseudo-first-order model. The degradation rate constant (19 x 10(-4)min(-1)) in the combined reaction with UV-C (4.2 mW cm(-2)) and Fe0 (5 mg L(-1)) was significantly enhanced compared to Fe0-only (4.8 x 10( 4) min(-1)) and photolytic reactions (2.25 x 10(-4)min(-1)), respectively. The removal efficiency of 1,4-dioxane in combined reaction with Fe0 and UV within 4 h was enhanced by increasing UV intensity at UV-C region (34% at 4.2 mW cm(-2) and 89% at 16.9 mW cm(-2)) comparing with the removal in the combined reaction with Fe0 and UV-A (29% at 2.1 mW cm(-2), and 33% at 12.6 mW cm(-2)). It indicates that 1,4-dioxane was degraded mostly by OH radicals in the combined reaction. The degradation patterns in both Fe(0)-only and combined reactions were well fitted to the Langmuir-Hinshelwood model, implying that adsorption as well as the chemical reaction occurred. The transformation of Fe0 to Fe2+ and Fe3+ was observed in the Fe0-only and combined reactions, and the transformation rate of Fe0 was improved by UV irradiation. Furthermore, the reduction of Fe3+ was identified in the combined reaction, and the reduction rate was enhanced by an increase of UV energy. Our study demonstrated that the enhancement of 1,4-dioxane removal rate occurred via an increased supply of OH radicals from the Fenton-like reaction induced by the photolysis of Fe0 and H2O, and with producing less sludge. PMID- 19131084 TI - Translation elongation factor 1A is a component of the tombusvirus replicase complex and affects the stability of the p33 replication co-factor. AB - Host RNA-binding proteins are likely to play multiple, integral roles during replication of plus-strand RNA viruses. To identify host proteins that bind to viral RNAs, we took a global approach based on the yeast proteome microarray, which contains 4080 purified yeast proteins. The biotin-labeled RNA probes included two distantly related RNA viruses, namely Tomato bushy stunt virus (TBSV) and Brome mosaic virus (BMV). Altogether, we have identified 57 yeast proteins that bound to TBSV RNA and/or BMV RNA. Among the identified host proteins, eleven bound to TBSV RNA and seven bound to BMV RNA with high selectivity, whereas the remaining 39 host proteins bound to both viral RNAs. The interaction between the TBSV replicon RNA and five of the identified host proteins was confirmed via gel-mobility shift and co-purification experiments from yeast. Over-expression of the host proteins in yeast, a model host for TBSV, revealed 4 host proteins that enhanced TBSV replication as well as 14 proteins that inhibited replication. Detailed analysis of one of the identified yeast proteins binding to TBSV RNA, namely translation elongation factor eEF1A, revealed that it is present in the highly purified tombusvirus replicase complex. We also demonstrate binding of eEF1A to the p33 replication protein and a known cis-acting element at the 3' end of TBSV RNA. Using a functional mutant of eEF1A, we provide evidence on the involvement of eEF1A in TBSV replication. PMID- 19131087 TI - Phosphate dynamics in an urban sewer: a case study of Nancy, France. AB - The nature of phosphate phases present in suspended matter, biofilm, and sediment of Greater Nancy sewer system was investigated over a period of two years. The phosphate speciation was determined by two approaches: a direct identification of phosphorus mineral phases was conducted by Transmission Electron Microscopy (TEM) coupled with energy-dispersive X-ray spectroscopy (EDXS), whereas a chemical extraction of samples provided an estimate of phosphorus pools defined by the fractionation scheme. Quantitative analysis of 1340 individual particles by TEM EDXS allowed to draw a picture of phosphate species distributions along the sewer system and over time. Amorphous Ca-phosphates (brushite, whitlockite, octacalcium phosphate, Mg-brushite, hydroxyapatite and carbapatite) were ubiquitous although brushite dominated upstream, and octacalcium phosphate and apatite prevailed downstream and in sediments. Al-Ca-phosphate minerals such as foggite, bearthite, gatumbaite, and crandallite appeared downstream and in biofilms. Ca-phosphate phase assemblages in the different locations of the sewer system were dependent on phase transformations from brushite to hydroxyapatite that were shown to be kinetically driven. The restriction of Al-Ca-phosphates to downstream of the sewer system was most probably related to the lower pHs measured at these sites. The pH dependency was confirmed by stability calculations. Chemical extractions were not reliable. TEM examination of extraction residues revealed the presence of neoformed Al-Ca-phosphate species that invalidated the fractionation scheme. Nonetheless, it confirmed that phosphate phases may undergo significant geochemical changes over a short time scale. PMID- 19131088 TI - Biological removal of phenol from strong wastewaters using a novel MSBR. AB - In this study, the performance of a moving-bed sequencing batch reactor (MSBR) that removes phenol from wastewater is presented. The effects of phenol concentration (50-3325 mg L(-1)), filling time (0-4 h) and aerating time (4-18 h) on the performance of the MSBR are given in terms of phenol and COD removal efficiencies. Moreover, the effect of the moving media on the overall performance of the reactor is also determined. The reactor can completely remove phenol and COD at inlet concentrations up to 3000 mg phenol L(-1) (6780 mg COD L(-1)), which was the inhibition concentration, and with a 24-h cycle time. The filling time range tested here did not significantly affect phenol or COD removal. The optimum hydraulic retention time (HRT) for the MSBR is 40 h and the critical phenol loading rate is 83.4 g phenol m(-3) h(-1), which gives a phenol removal efficiency of 99%. The reactor can also withstand shock loads from slug feeding. The moving bed contribution to phenol and COD removal efficiencies was up to 28.1 and 34.7%, respectively, at the phenol loading rate of 83.4 g m(-3) h(-1). The findings of this investigation suggest that MSBR can be a robust and promising process for effectively treating wastewaters containing inhibitor or recalcitrant compounds in industrial settings. PMID- 19131089 TI - Absorbance spectroscopy-based examination of effects of coagulation on the reactivity of fractions of natural organic matter with varying apparent molecular weights. AB - Absorbance spectra of fractions of natural organic matter (NOM) with varying apparent molecular weights (AMWs) were examined in this study. Size exclusion chromatography (SEC) was employed to obtain AMW distributions for three Australian water sources which represented low- and high-dissolved organic carbon (DOC) surface waters and a source with highly degraded NOM. These waters were coagulated with alum and other coagulants. Effects of coagulation on AMW distributions were quantified based on an absorbance slope index (ASI) calculated using NOM absorbance measured at 220, 230, 254 and 272 nm. This index can be calculated for any AMW fraction of NOM. Similarly to SUVA(254), ASI values decrease consistently in coagulated waters and are correlated with trihalomethane yields. Comparison of ASI indexes in different water sources indicates the presence of both common trends and differences indicative of NOM site specificity. PMID- 19131090 TI - Decomposition and mineralization of aquatic humic substances (AHS) in treating landfill leachate using the Anammox process. AB - Aquatic humic substance (AHS) in landfill leachate is resistant to biodegradation, especially in anaerobic habitats. However, we reported here for the first time that AHS was completely biodegraded with the anaerobic ammonium oxidation (Anammox) process. As a result of aromatic-ring cleavage and mineralization, carboxylic and aliphatic organics were significantly produced, the contents of chromophores such as quinoid and ketone were remarkably decreased to decolorize the leachate, and produced carbon dioxide along with increasing carbonate was clearly presented. Of the degraded AHS of 137 mg L(-1), 51 mg L(-1) was owed to the oxidation with sulfate of 76 mg L(-1) as electron acceptor, and the rest to other metabolic mechanism. Isolation and identification of heterotrophic bacteria revealed a diversified consortium comprising four facultative anaerobic species, Bacillus sp., Paenibacillus sp., Bacteroides sp. and Staphylococcus sp., without sulfate-reducing bacteria detected. Their contribution to AHS biodegradation and sulfate reduction under the special conditions with high oxidization-reduction potential and insufficient electron acceptors has not been known yet. Further work is underway to investigate their properties and respective duties in the consortium. PMID- 19131091 TI - Determination of lead speciation in melting furnace fly ash by sequential chemical extraction. AB - A procedure for sequential chemical extraction to determine the degree of speciation of Pb compounds in melting furnace fly ash was developed. Optimal solvent concentrations were determined individually for each extracting solvent by using standard Pb reagents. Sequential chemical extraction was performed to evaluate the selectivity of extraction procedures for each Pb compound using simulated melting furnace fly ash samples. The extraction of Pb was significantly inhibited by the formation of insoluble precipitates created by the combination of extracted Pb(2+) with SO(4)(2-), Cl(-), and OH(-) contained in melting furnace fly ash. To selectively extract each Pb compound, Pb(2+) was sulfurized to prevent the formation of insoluble precipitates, and the solution pH was controlled to neutralize OH(-). Consequently, by the parallel use of five extraction procedures, the concentration of Pb in simulated melting furnace fly ash samples was satisfactorily estimated with errors between 0.5% and 20%. PMID- 19131092 TI - Risk from exposure to trihalomethanes during shower: probabilistic assessment and control. AB - Exposure to trihalomethanes (THMs) through inhalation and dermal contact during showering and bathing may pose risks to human health. During showering and bathing, warm water (35 degrees C-45 degrees C) is generally used. Warming of chlorinated supply water may increase THMs formation through enhanced reactions between organics and residual chlorine. Exposure assessment using THMs concentrations in cold water may under-predict the possible risks to human health. In this study, THMs concentrations in warm water were estimated by developing a THMs formation rate model. Using THMs in warm water, cancer and non cancer risks to human health were predicted for three major cities in Ontario (Canada). The parameters for risk assessments were characterized by statistical distributions. The total cancer risks from exposure to THMs during showering were predicted to be 7.6x10(-6), 6.3x10(-6) and 4.3x10(-6) for Ottawa, Hamilton and Toronto respectively. The cancer risks exceedance probabilities were estimated to be highest in Ottawa at different risk levels. The risks through inhalation exposure were found to be comparable (2.1x10(-6)-3.7x10(-6)) to those of the dermal contact (2.2x10(-6)-3.9x10(-6)) for the cities. This study predicted 36 cancer incidents from exposure to THMs during showering for these three cities, while Toronto contributed the highest number of possible cancer incidents (22), followed by Ottawa (10) and Hamilton (4). The sensitivity analyses showed that health risks could be controlled by varying shower stall volume and/or shower duration following the power law relationship. PMID- 19131093 TI - Endothelin is not elevated in acute pulmonary embolism. AB - INTRODUCTION: In acute pulmonary embolism (APE) the increase of pulmonary vascular resistance depends on the thromboli load and potentially on the pulmonary bed contraction caused by neurohormonal reaction. Plasma levels of endothelin were reported to be elevated in pulmonary arterial hypertension. However, there are only a few studies assessing endothelin in patients with APE. MATERIALS & METHODS: Therefore in our study we evaluated endothelin concentration in 55 patients (29M, 26F, age 57+/-19 yrs) with confirmed APE for potential value in risk stratification. Patients were compared with 24 healthy volunteers at similar age. On admission blood samples were collected for plasma endothelin concentration. The quantitative assessment of right ventricular (RV) function was performed by echocardiography. RESULTS: Endothelin concentrations were similar in APE patients and in control group (1.41(0.22-9.68)pg/mL vs. 1.62(0.27-8.92)pg/mL; p = NS). There was no differences in endothelin levels between APE patients with and without RV dysfunction (1.46(0.38-4.54)pg/mL vs. 1.41(0.22-9.68)pg/mL; p = NS). Endothelin concentration did not differ between patients with serious adverse events and APE group with event-free clinical course (3.19(0.38 4.27)pg/mL vs. 1.38(0.22-9.68)pg/mL; p = NS). There was no significant correlation between endothelin levels and blood saturation, time from the first symptoms, heart rate, blood pressure, tricuspid valve regurgitation pressure gradient and other echocardiographic parameters. CONCLUSIONS: We concluded that plasma endothelin concentrations assessed on admission are not elevated in patients with APE and it does not play as important role in acute phase of increase of pressure in pulmonary arteries as in chronic pulmonary hypertension. PMID- 19131094 TI - Abdominal radical trachelectomy: Success and pitfalls in a general gynecologic oncology practice. AB - OBJECTIVE: To report our successes and complications with a series of abdominal radical trachelectomies performed to preserve fertility in young women at the Massachusetts General Hospital (MGH). METHODS: Institutional review board (IRB) permission was obtained for retrospective record review. Data were collected regarding patient age and parity, tumor stage and histology, surgical time and complications, post-operative complications, follow-up, and pregnancy. RESULTS: Ten patients underwent radical abdominal trachelectomy, 9 by the same surgeon (LD). Surgery was essentially identical to that of radical hysterectomy with the exception of re-anastomosis of the uterine fundus to the vagina and placement of cerclage. Pre-operative evaluation and post operative follow-up was for the most part identical for all patients. Two patients achieved pregnancy, with 1 twin delivery and 1 patient had 2 pregnancies. Two patients experienced cervical stenosis with regular menses and the same 2 patients passed their abdominal cerclage vaginally. To date there have been no cancer recurrences. Pap smear follow-up has been complicated by difficulty in reading smears from the lower uterine segment (LUS). CONCLUSION: Radical abdominal trachelectomy can be successfully performed by any gynecologic oncologist who is trained in radical pelvic surgery. Pre-operative counseling is crucial in obtaining informed consent. Patients must be aware of potential post-operative complications, including pre-term delivery. Cytology department needs to be aware of potential pitfalls in reading Pap smears from the LUS. PMID- 19131095 TI - Plasma luteinizing hormone concentrations in cows given repeated treatments or three different doses of gonadotropin releasing hormone. AB - We hypothesized that: (i) repeated GnRH treatments would increase the magnitude and duration of the LH surge and would increase progesterone (P4) concentrations after ovulation; and (ii) the release of pituitary LH would be greater in response to larger doses of GnRH. In Experiment 1, ovary-intact cows were given an intravaginal P4 (1.9g) insert (CIDR) for 10 d and 500 microg cloprostenol (PGF) at CIDR removal to synchronize estrus. On Days 7 or 8 after estrus, cows received two PGF treatments (12h apart) and 100 microg GnRH at 36 (Control), 36 and 38 (GnRH38), or 36 and 40h (GnRH40) after the first PGF. Mean plasma LH concentration (ng/mL) was greater (P<0.05) in GnRH38 (8.8+/-1.1) than in Control (5.1+/-1.3), with that in GnRH40 (5.8+/-1.3) being intermediate. Although the duration (h) of the LH surge was longer in GnRH40 (8.0+/-0.4) than in either GnRH38 (P<0.05; 7.0+/-0.3) or Control (P<0.09; 7.1+/-0.4), mean postovulatory P4 (ng/mL) was greater (P<0.01) in Control (4.2+/-0.7) than in GnRH38 (2.9+/-0.6) or GnRH40 (3.0+/-0.7) cows. In Experiment 2, ovariectomized cows were given a CIDR for 10 d and 2mg of estradiol cypionate im at CIDR insertion. Thirty-six hours after CIDR removal, cows received, 50, 100, or 250 microg of GnRH. Cows given 250 microg GnRH released more LH (9.4+/-1.4ng/mL) than those given 50 or 100 microg (6.1+/-1.3 and 5.4+/-1.4ng/mL, respectively), and had an LH surge of longer duration than those given 50 microg (6.8+/-0.4 vs. 5.1+/-0.3h). In summary, ovary intact cows in the GnRH38 group had greater mean and peak LH concentrations, but subsequent plasma P4 concentrations were lower than in Control cows. Ovariectomized cows given 250 microg GnRH had a greater pituitary release of LH. PMID- 19131096 TI - Effect of porous glass-ceramic fillers on mechanical properties of light-cured dental resin composites. AB - OBJECTIVES: The aim of this work was to study the effect of porous particles on the mechanical properties of dental resin composites. METHODS: Two kinds of glass ceramic powders with different calcium-mica to fluorapatite ratio were used as inorganic fillers for light-cured dental resin composites. The glass-ceramic particles were etched to introduce porous structures. The microstructures of particles and the fractography of dental composites were observed using SEM/EDS. The crystalline phases in the fillers were determined by XRD. The specific surface areas (SBET) of porous fillers were tested to show the degree of porosity. 6 samples were prepared for each bending strength test, which was conducted in a Mechanical Testing System. RESULTS: The results show that the bending strength does not vary monotonically with the porosity of the particles. For the glass-ceramics with a high nominal calcium-mica to fluorapatite ratio, the dental resin filled with porous particles has a higher strength than that filled with dense particles; however, for the glass-ceramics with a low nominal calcium-mica to fluorapatite ratio, the dental resin filled with porous particles has a lower strength. SIGNIFICANCE: The study indicates that the porosity itself is not a decisive factor for strengthening of dental resin composites. The composition of the glass-ceramic particles and the porous structure are also essential factors influencing the mechanical strength. The investigation on the effect of porous glass-ceramic fillers will provide more understanding on the mechanical properties of dental resin composites and will help with the design of new dental restorative materials. PMID- 19131097 TI - Measles in Europe: an epidemiological assessment. AB - BACKGROUND: Measles persists in Europe despite the incorporation of the measles vaccine into routine childhood vaccination programmes more than 20 years ago. Our aim was therefore to review the epidemiology of measles in relation to the goal of elimination by 2010. METHODS: National surveillance institutions from 32 European countries submitted data for 2006-07. Data for age-group, diagnosis confirmation, vaccination, hospital treatment, the presence of acute encephalitis as a complication of disease, and death were obtained. 30 countries also supplied data about importation of disease. Clinical, laboratory-confirmed, and epidemiologically linked cases that met the requirements for national surveillance were analysed. Cases were separated by age: younger than 1 year, 1-4 years, 5-9 years, 10-14 years, 15-19 years, and older than 20 years. Countries with indigenous measles incidence per 100 000 inhabitants per year of 0, less than 0.1, 0.1-1, and more than 1 were grouped into categories of zero, low, moderate, and high incidence, respectively. FINDINGS: For the 2 years of the study, 12 132 cases of measles were recorded with most cases (n=10 329; 85%) from five countries: Romania, Germany, UK, Switzerland, and Italy. Most cases were unvaccinated or incompletely vaccinated children; however, almost a fifth were aged 20 years or older. For the same 2 years, seven measles-related deaths were recorded. High measles incidence in some European countries revealed suboptimum vaccination coverage. Of the 210 cases that were reported as being imported, 117 (56%) came from another country within Europe and 43 (20%) from Asia. INTERPRETATION: The suboptimum vaccination coverage raises serious doubts that the goal of elimination by 2010 can be attained. Achievement and maintenance of optimum vaccination coverage and improved surveillance are the cornerstones of the measles elimination plan for Europe. PMID- 19131098 TI - Measles in Europe--there is room for improvement. PMID- 19131099 TI - Clam dredging effects and subsequent recovery of benthic communities at different depth ranges. AB - This study aimed to assess the potential effects of clam dredging and the subsequent recovery of the benthic environment. Two experimental areas located at 6 and 18m depth were established in order to analyse whether impacts and recovery of benthic environment are depth-related. Study areas were located within an area closed to dredging and two different plots were established at both depths. One of the plots was subjected to intense clam dredge-fishing, while the other was undisturbed and therefore used as control. Sampling followed a BACI design, with samples for macrobenthic, meiobenthic and sediment particle size analysis being taken by SCUBA divers from both areas before and after fishing stress. For both depths, impacts on the benthic environment were very low resulting in high recovery rates. Nevertheless, at shallower depths communities demonstrated a faster recovery. It was shown that depending on the faunal component used as a bioindicator, different results can be observed. Generally deposit-feeding organisms with scales or chitinous plates and vermiform shape (mainly crustaceans, polychaetes and ophiuroids), without external protection, were the most affected by dredging, while some polychaetes without external protection and with a carnivorous feeding mode seemed to be enhanced by fishing. The low level of perturbations induced by the dredging activities was comparable to the impact of surface waves on the bottom, as experiments were undertaken in wave-dominated environments. The coexistence of storm events during the study period proved to have similar or even more deleterious effects on the benthic environment. It appears that communities from hydrodynamic fishing grounds that are well adapted to natural physical stress are not highly affected by dredging. PMID- 19131100 TI - Sunken wood habitat for thiotrophic symbiosis in mangrove swamps. AB - Large organic falls to the benthic environment, such as dead wood or whale bones, harbour organisms relying on sulfide-oxidizing symbionts. Nothing is known however, concerning sulfide enrichment at the wood surface and its relation to wood colonization by sulfide-oxidizing symbiotic organisms. In this study we combined in situ hydrogen sulfide and pH measurements on sunken wood, with associated fauna microscopy analyses in a tropical mangrove swamp. This shallow environment is known to harbour thiotrophic symbioses and is also abundantly supplied with sunken wood. A significant sulfide enrichment at the wood surface was revealed. A 72h sequence of measurements emphasized the wide fluctuation of sulfide levels (0.1->100muM) over time with both a tidal influence and rapid fluctuations. Protozoans observed on the wood surface were similar to Zoothamnium niveum and to vorticellids. Our SEM observations revealed their association with ectosymbiotic bacteria, which are likely to be sulfide-oxidizers. These results support the idea that sunken wood surfaces constitute an environment suitable for sulfide-oxidizing symbioses. PMID- 19131101 TI - Polymeric drugs based on bioactive glycosides for the treatment of brain tumours. AB - Polymeric drugs carrying glycolipids have been designed as target macromolecules for the treatment of brain tumours. A methacrylate derivative of oleyl 2 acetamido-2-deoxy-alpha-D-glucopyranoside (OAGMA) has been prepared and the corresponding glycopolymer obtained by free radical polymerisation. To modulate the hydrophobic character of the polymeric drug, the acrylic glycomonomer was copolymerised with vinyl pyrrolidone (VP). Reactivity ratios obtained by performing copolymerisation reactions inside the NMR apparatus were r(OAGMA)=5.94 and r(VP)=0.01, indicating the much higher reactivity of the glycomonomer. The hydrolytical release of oleyl 2-acetamido-2-deoxy-alpha-D-glucopyranoside (OAG) from the copolymeric drugs was produced in vitro by the ester enzymatic hydrolysis using enzyme/buffered solutions. The cytotoxicity of OAG and OAGMA tested against a human glioblastoma line and normal fibroblasts revealed a concentration dependent selectivity towards tumour cells versus fibroblasts. The antimitotic activity of the copolymeric drugs was also confirmed. The addition of the eluates of the copolymeric systems collected at 1 and 2 days produced a significant decrease in cellular viability of the glioblastoma cells without affecting that of normal fibroblasts. On the contrary, fibroblasts were able to adhere and proliferate onto the copolymeric systems showing normal morphology and revealing a good biocompatibility of the copolymeric drugs against healthy cells. PMID- 19131102 TI - Enhanced bioactivity of bone morphogenetic protein-2 with low dose of 2-N, 6-O sulfated chitosan in vitro and in vivo. AB - Bone morphogenetic protein-2 (BMP-2) has been widely used as an effective growth factor in bone tissue engineering. However, large amounts of BMP-2 are required to induce new bone and the resulting side effects limit its clinical application. Sulfated polysaccharides, such as native heparin, and heparan sulfate have been found to modulate BMP-2 bioactivity and play pivotal roles in bone metabolism. Whereas the direct role of chitosan modified with sulfate group in BMP-2 signaling has not been reported till now. In the present study, several sulfated chitosans with different positions were synthesized by regioselective reactions firstly. Using C2C12 myoblast cells as in vitro models, the enhanced bioactivity of BMP-2 was attributed primarily to the stimulation from 6-O-sulfated chitosan (6SCS), while 2-N-sulfate was subsidiary group with less activation. Low dose of 2-N, 6-O-sulfated chitosan (26SCS) showed significant enhancement on the alkaline phosphatase (ALP) activity and the mineralization formation induced by BMP-2, as well as the expression of ALP and osteocalcin mRNA. Moreover, increased chain length and further sulfation on 26SCS also resulted in a higher ALP activity. Dose-dependent effects on BMP-2 bioactivity were observed in both sulfated chitosan and heparin. Compared with native heparin, 26SCS showed much stronger simultaneous effects on the BMP-2 bioactivity at low dose. Stimulated secreted Noggin protein failed to block the function of BMP-2 in the presence of 26SCS. The BMP-2 ligand bound to its receptor was enhanced by low dose of 26SCS, whereas weakened by the increasing amounts of 26SCS. Furthermore, simultaneous administration of BMP-2 and 26SCS in vivo dose-dependently induced larger amounts of ectopic bone formation compared with BMP-2 alone. These findings clearly indicate that 26SCS is a more potent enhancer for BMP-2 bioactivity to induce osteoblastic differentiation in vitro and in vivo by promoting BMP-2 signaling pathway, suggesting that 26SCS could be used as the synergistic factor of BMP-2 for bone regeneration. PMID- 19131104 TI - Transcriptomic analysis of polyamine-related genes and polyamine levels in placenta, yolk sac and fetus during the second half of mouse pregnancy. AB - In mammals, polyamines are essential for the maintenance of cell growth. Although early studies reported the highest values of mammalian ornithine decarboxylase (ODC) activity, a key enzyme in polyamine biosynthesis, in rodent placenta, the role of this enzyme in the second half of rodent pregnancy is still controversial. In order to get new insights on polyamine metabolism during this period of pregnancy, we studied polyamine levels, ODC expression and activity and transcript profile of different polyamine-related genes in mouse placenta, fetus and yolk sac. Results indicated that ODC activity and protein levels were higher in placenta than in fetus and yolk sac, especially in the labyrinth, although no correlation between ODC activity and polyamine levels were observed. The half life of placental ODC ( approximately 190 min) was also higher than the fetal one ( approximately 24 min). Messenger RNAs of all biosynthetic and retroconversion enzymes of polyamine metabolism were present in the three gestational compartments analyzed, as well as those of antizymes 1 and 2 and antizyme inhibitor 1. However, no expression of antizyme 3 and antizyme inhibitor 2 was detected. The catabolic enzyme diamine oxidase was expressed only in the maternal part of placenta but not in the fetal part or in the fetus. The expansion of polyamine pools in the fetus was markedly higher than in placenta, in spite of its lower biosynthetic activity. Our results suggest that the elevated polyamine biosynthetic activity of mouse placenta is required to satisfy the high demand of polyamines required by the growing fetus, during the later period of pregnancy. PMID- 19131103 TI - Influence of anchoring ligands and particle size on the colloidal stability and in vivo biodistribution of polyethylene glycol-coated gold nanoparticles in tumor xenografted mice. AB - Polyethylene glycol (PEG)-coated (pegylated) gold nanoparticles (AuNPs) have been proposed as drug carriers and diagnostic contrast agents. However, the impact of particle characteristics on the biodistribution and pharmacokinetics of pegylated AuNPs is not clear. We investigated the effects of PEG molecular weight, type of anchoring ligand, and particle size on the assembly properties and colloidal stability of PEG-coated AuNPs. The pharmacokinetics and biodistribution of the most stable PEG-coated AuNPs in nude mice bearing subcutaneous A431 squamous tumors were further studied using (111)In-labeled AuNPs. AuNPs coated with thioctic acid (TA)-anchored PEG exhibited higher colloidal stability in phosphate buffered saline in the presence of dithiothreitol than did AuNPs coated with monothiol-anchored PEG. AuNPs coated with high-molecular-weight (5000 Da) PEG were more stable than AuNPs coated with low-molecular-weight (2000 Da) PEG. Of the 20-nm, 40-nm, and 80-nm AuNPs coated with TA-terminated PEG(5000), the 20-nm AuNPs exhibited the lowest uptake by reticuloendothelial cells and the slowest clearance from the body. Moreover, the 20-nm AuNPs coated with TA-terminated PEG(5000) showed significantly higher tumor uptake and extravasation from the tumor blood vessels than did the 40- and 80-nm AuNPs. Thus, 20-nm AuNPs coated with TA-terminated PEG(5000) are promising potential drug delivery vehicles and diagnostic imaging agents. PMID- 19131105 TI - Adhesion molecules in human trophoblast - a review. II. extravillous trophoblast. AB - At the tips of anchoring villi, cytotrophoblast (CTB) proliferation leads to a process of multilayering in which cells lose their attachment to the villous basement membrane and develop into columns, within which they adhere to one another using desmosomes, with associated intermediate filament bundles. Non desmosomal cadherins, tight junction proteins and other adhesion molecules are also present, suggesting that actin-associated adhesions contribute to placental anchorage. In the distal columns, cell-cell interactions diminish, cells upregulate beta1 integrins and bind to a provisional fibrinoid extracellular matrix, eventually detaching to migrate into the decidual stroma and myometrium, where interstitial and endovascular extravillous trophoblast (EVT) populations show distinct repertoires of adhesion molecules. PMID- 19131106 TI - Adhesion molecules in human trophoblast - a review. I. Villous trophoblast. AB - In the placental villus, cells attach to basement membrane via integrin alpha6beta4 and adhere both laterally and apically to their neighbours. The most prominent adhesive specialisation seen using the electron microscope is the desmosome, which connects cytotrophoblast cells (CTB) laterally and also contributes to the attachment of CTB to the overlying syncytium. However, numerous cadherins and other junctional proteins are also present in the corresponding plasma membrane domains, indicating a multiplicity of adhesive interactions. Integrins, tight junction components and cadherins are all found in the syncytial microvillous membrane, perhaps reflecting its ability to form intersyncytial bridges. There is a wide gulf to be filled between molecular anatomy and functional studies, with much to be learned about the role of adhesion molecules in regulating villous epithelial integrity, homeostasis and growth. PMID- 19131107 TI - Intracellular calcium signals regulate growth of hepatic stellate cells via specific effects on cell cycle progression. AB - Hepatic stellate cells (HSC) are important mediators of liver fibrosis. Hormones linked to downstream intracellular Ca(2+) signals upregulate HSC proliferation, but the mechanisms by which this occurs are unknown. Nuclear and cytosolic Ca(2+) signals may have distinct effects on cell proliferation, so we expressed plasmid and adenoviral constructs containing the Ca(2+) chelator parvalbumin (PV) linked to either a nuclear localization sequence (NLS) or a nuclear export sequence (NES) to block Ca(2+) signals in distinct compartments within LX-2 immortalized human HSC and primary rat HSC. PV-NLS and PV-NES constructs each targeted to the appropriate intracellular compartment and blocked Ca(2+) signals only within that compartment. PV-NLS and PV-NES constructs inhibited HSC growth. Furthermore, blockade of nuclear or cytosolic Ca(2+) signals arrested growth at the G2/mitosis (G2/M) cell-cycle interface and prevented the onset of mitosis. Blockade of nuclear or cytosolic Ca(2+) signals downregulated phosphorylation of the G2/M checkpoint phosphatase Cdc25C. Inhibition of calmodulin kinase II (CaMK II) had identical effects on LX-2 growth and Cdc25C phosphorylation. We propose that nuclear and cytosolic Ca(2+) are critical signals that regulate HSC growth at the G2/M checkpoint via CaMK II-mediated regulation of Cdc25C phosphorylation. These data provide a new logical target for pharmacological therapy directed against progression of liver fibrosis. PMID- 19131108 TI - Alternative splicing of RyR1 alters the efficacy of skeletal EC coupling. AB - Alternative splicing of ASI residues (Ala(3481)-Gln(3485)) in the skeletal muscle ryanodine receptor (RyR1) is developmentally regulated: the residues are present in adult ASI(+)RyR1, but absent in the juvenile ASI(-)RyR1 which is over expressed in adult myotonic dystrophy type 1 (DM1). Although this splicing switch may influence RyR1 function in developing muscle and DM1, little is known about the properties of the splice variants. We examined excitation-contraction (EC) coupling and the structure and interactions of the ASI domain (Thr(3471) Gly(3500)) in the splice variants. Depolarisation-dependent Ca(2+) release was enhanced by >50% in myotubes expressing ASI(-)RyR1 compared with ASI(+)RyR1, although DHPR L-type currents and SR Ca(2+) content were unaltered, while ASI( )RyR1 channel function was actually depressed. The effect on EC coupling did not depend on changes in ASI domain secondary structure. Probing RyR1 function with peptides possessing the ASI domain sequence indicated that the domain contributes to an inhibitory module in RyR1. The action of the peptide depended on a sequence of basic residues and their alignment in an alpha-helix adjacent to the ASI splice site. This is the first evidence that the ASI residues contribute to an inhibitory module in RyR1 that influences EC coupling. Implications for development and DM1 are discussed. PMID- 19131109 TI - cADPR stimulates SERCA activity in Xenopus oocytes. AB - The intracellular second messenger cyclic ADP-ribose (cADPR) induces Ca(2+) release through the activation of ryanodine receptors (RyRs). Moreover, it has been suggested that cADPR may serve an additional role to modulate sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) pump activity, but studies have been complicated by concurrent actions on RyR. Here, we explore the actions of cADPR in Xenopus oocytes, which lack RyRs. We examined the effects of cADPR on the sequestration of cytosolic Ca(2+) following Ca(2+) transients evoked by photoreleased inositol 1,4,5-trisphosphate (InsP(3)), and by Ca(2+) influx through expressed nicotinic acetylcholine receptors (nAChR) in the oocytes membrane. In both cases the decay of the Ca(2+) transients was accelerated by intracellular injection of a non-metabolizable analogue of cADPR, 3-Deaza-cADPR, and photorelease of cADPR from a caged precursor demonstrated that this action is rapid (a few s). The acceleration was abolished by pre-treatment with thapsigargin to block SERCA activity, and was inhibited by two specific antagonists of cADPR, 8-NH(2)-cADPR and 8-br-cADPR. We conclude that cADPR serves to modulate Ca(2+) sequestration by enhancing SERCA pump activity, in addition to its well-established action on RyRs to liberate Ca(2+). PMID- 19131110 TI - Compound 48, a novel dual PPAR alpha/gamma ligand, inhibits the growth of human CML cell lines and enhances the anticancer-effects of imatinib. AB - Compound 48 (C48) is a novel dual ligand for peroxisome proliferator-activated receptor alpha and gamma (PPAR alpha/gamma). Culture of imatinib-sensitive and resistant CML cell lines with C48 resulted in a strong growth inhibition which associated with G0/G1 cell cycle arrest. However, it showed no obvious toxicity to normal CD34(+) hematopoietic stem cells. Decrease of pSTATs and pAKT were noticed suggesting that interference of AKT and STATs signaling may be the mechanisms for the effects of PPAR alpha/gamma ligands. Of more clinical importance, this ligand strongly enhanced the anticancer-effects of imatinib. Overall, our data suggest that the PPAR alpha/gamma ligands may have potentials in the treatment of CML in an adjuvant setting either before or after the development of imatinib resistance. PMID- 19131111 TI - Risk factors for bacteremia during and after adenoidectomy and/or adenotonsillectomy. AB - OBJECTIVE: To evaluate the incidence and persistence of bacteremia in children undergoing adenoidectomy or adenotonsillectomy for different medical reasons. METHODS: We enrolled 130 children scheduled for adenoidectomy because of recurrent acute otitis media (rAOM, 15) or persistent otitis media with effusion (pOME, 33), or for adenotonsillectomy because of obstructive sleep apnea syndrome (OSAS, 41) or recurrent tonsillopharyngitis (rTF, 41). Nasopharyngeal aspirates taken just before surgery, swabs of the ablated central adenoidal and tonsillar tissues, and blood samples taken within the first 30s of beginning the operation and 20min after its end were used for bacterial cultures. RESULTS: The incidence of positive blood cultures after the beginning of the operation was significantly higher in the children who underwent adenotonsillectomy than in those who underwent adenoidectomy, and in those with rAOM or rTF than in those with pOME or OSAS. Children with nasopharyngeal colonisation were significantly more likely to have a positive blood culture than those without. Twenty of the 25 children with a positive blood culture (80.0%), had the same bacteria in their nasopharyngeal and adenoidal/tonsillar tissues. CONCLUSIONS: Our results show that bacteremia is significantly more frequently associated with adenotonsillectomy than with adenoidectomy, and significantly more frequent in patients with a history of rAOM or rTF. PMID- 19131113 TI - A GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitis. AB - Quantitative real time PCR (Q-PCR) is the method of choice to study mRNA expression levels. Since Q-PCR is very sensitive, normalization of the data with stably expressed reference genes if of utmost importance. The stability of reference genes depends on the tissue and the species of interest. Therefore, evaluation of the stability of reference genes must be performed for each new tissue and species under study. The stability of B2M, GAPDH, HPRT, SRPR, hnRNPH, GUSB, RPL8, RPS5, and RPS19 was analyzed with the GeNorm software in snap frozen canine skin biopsies. Healthy dogs (n=7) and dogs with confirmed atopic dermatitis (n=28) were included. Lesional and non-lesional skin was analyzed. The study indicated that the most appropriate reference genes in canine skin are the ribosomal gene products RPL8, RPS5 and RPS19 besides GUSB and HPRT. As little as three reference genes will reveal highly reliable Q-PCR calculations. PMID- 19131112 TI - Persistent versus transient depressive symptoms in relation to platelet hyperactivation: a longitudinal analysis of dementia caregivers. AB - BACKGROUND: Depressive symptoms and caregiving stress may contribute to cardiovascular disease (CVD) via chronic platelet activation; however, it remains unclear whether this elevated activation constitutes a trait or state marker. The primary objective was to investigate whether persistent depressive symptoms would relate to elevated platelet activation in response to acute psychological stress over a three-year period. METHODS: Depressive symptoms (Brief Symptom Inventory) were assessed among 99 spousal dementia caregivers (52-88 years). Platelet P selectin expression was assessed in vivo using flow cytometry at three time points over the course of an acute stress test: baseline, post-stress, and after 14 min of recovery. Two competing structural analytic models of depressive symptoms and platelet hyperactivity with three yearly assessments were compared. RESULTS: Although depressive symptoms were generally in the subclinical range, their persistent elevation was associated with heightened platelet reactivity and recovery at all three-years while the change in depressive symptoms from the previous year did not predict platelet activity. LIMITATIONS: These results focus on caregivers providing consistent home care, while future studies may extend these results by modeling major caregiving stressors. CONCLUSIONS: Enduring aspects of negative affect, even among those not suffering from clinical depression are related to hemostatic changes, in this case platelet reactivity, which might be one mechanism for previously reported increase in CVD risk among elderly Alzheimer caregivers. PMID- 19131114 TI - Toll-like receptors TLR1, TLR2 and TLR4 gene mutations and natural resistance to Mycobacterium avium subsp. paratuberculosis infection in cattle. AB - Toll like receptors (TLRs) are a class of pattern recognition receptors belonging to the innate immune system. Mutations in the protein coding region of TLRs are associated with altered responsiveness to pathogen-associated molecular patterns (PAMPs). A search was performed for novel mutations in bovine TLR1, TLR2 and TLR4 genes associated with the Mycobacterium avium subsp. paratuberculosis (MAP) infection. The work was also focused on the assessment of linkage between well known mutations in TLR genes (TLR2: Arg677Trp, Pro681His and Arg753Gln; TLR4: Asp299Gly and Thr399Ile), and the susceptibility of cattle to MAP infection. Detection of MAP infection in cattle population (n=711) was based on IS900 PCR, which revealed 22.50% (n=160) MAP positivity. Known mutations in TLR2 and TLR4 genes were not found in cattle population. A novel mutation Val220Met was associated (Odd's ratio, OR-3.459) with increased susceptibility to MAP infection. Toll/interleukin-1 receptor (TIR) domain of TLR2 was screened for the presence of mutations, wherein a novel Ile680Val mutation was linked with MAP infection. In silico analysis of the bovine TLR4 ectodomain (ECD) revealed the polymorphic nature of the central ECD and irregularities in the central LRR motifs. LRR11 of the TLR4 showed five missense mutations possibly linked with the increased susceptibility to MAP infection. The most critical position that may alter the pathogen recognition of TLR molecule was 4th residue downstream to LRR domain. Two such missense mutations in TLR4 (Asp299Asn downstream to LRR11, and Gly389Ser downstream to LRR15) were associated with MAP infection. Briefly, the work describes novel mutations in the bovine TLRs and presents their association with the MAP infection. PMID- 19131115 TI - Gene expression profiles of European wild boar naturally infected with Mycobacterium bovis. AB - Global gene expression profiles were analyzed in European wild boar naturally infected with Mycobacterium bovis. Spleen RNA was extracted from 23 M. bovis infected and 17 uninfected animals and analyzed using a Pigoligoarray representing 20,400 genes. Differentially expressed sequences (N=161) were identified affecting cellular processes such as apoptosis, cell communication and signal transduction, cell growth and/or maintenance, cytoskeleton organization and biogenesis, DNA repair, immune response, metabolism and energy pathways, protein metabolism, regulation of cell proliferation, regulation of gene expression, regulation of nucleic acid metabolism, regulation of physiological processes, and transport. Real-time RT-PCR analysis of mRNA levels was used to corroborate microarray results of selected genes. Immune response genes were among the most represented differentially expressed sequences and were selected for further discussion. Beta-defensin 129, T-cell surface glycoprotein CD8 and B cell receptor-associated protein 29 were overexpressed in infected animals. Lower expression levels of the immune response genes galectin-1, complement component C1qB and certain HLA class I and class II histocompatibility antigens and immunoglobulin chains were found in infected animals. This study identified new mechanisms by which naturally infected European wild boar respond to M. bovis infection and how the pathogen circumvents host immune responses to establish infection. Gene expression studies in naturally infected wildlife reservoirs of bovine tuberculosis are important for functional genomics and vaccine studies to aid in disease control in wildlife. PMID- 19131117 TI - Complete sequencing of full-length canine ataxia telangiectasia mutated mRNA and characterization of its putative promoter. AB - Ataxia telangiectasia mutated (ATM) protein is considered a "caretaker" of the genome integrity and a defective ATM has been correlated with increased cancer risk in human beings. In an effort to explore the reliability of dog as a spontaneous animal model of genetic susceptibility to lymphoid malignancies, we have carried out the complete sequencing of the canine ATM mRNA. 5' RACE analysis and sequencing were used to obtain the full-length canine cDNA. The transcription start site was found at CFA5: 27307661 (Dog Genome assembly 2.0, release 49). Two exons were found in the 5'UTR. A putative TATA-less bi-directional promoter region was found in the region 5' upstream of the cap site. The core promoter harbours different conserved regulatory motifs: CREB, CCAAT boxes (NF-binding sites), Sp1, AP-2, GCF, XRE, Ets, Cre and c-Myb. The major ORF, corresponding to the ortholog human and pig ATM isoform 1, has 64 exons and codes a protein of 3056 aa. The homology between dog and human ATM at the aa level was 89% identities-93% positives, even higher than the homology between pig and human. When compared with the canine genomic sequences, 3 sequence variants yielding to aa substitution were found. Canine ATM is highly conserved and may represent a candidate gene to evaluate lymphoid malignancies predisposition in dogs. PMID- 19131116 TI - Cloning and expression of ADAM-related metalloproteases in equine laminitis. AB - Equine laminitis is a debilitating disease affecting the digital laminae that suspend the distal phalanx within the hoof. While the clinical progression of the disease has been well documented, the molecular events associated with its pathogenesis remain largely unknown. Using real time quantitative PCR (RT-qPCR), we have investigated the expression of genes coding for proteins containing a Disintegrin and Metalloprotease domain (ADAM), as well as genes encoding the natural inhibitors of these enzymes (tissue inhibitor of metalloprotease; TIMP) in horses with naturally-acquired (acute, chronic and aggravated chronic clinical cases) or experimentally-induced (black walnut extract (BWE) and starch gruel models) laminitis. Changes in expression of these enzymes and regulators may underlie the pathologic remodeling of lamellar tissue in laminitis. Genes encoding ADAMs involved in inflammation (ADAM-10 and ADAM-17), as well as those implicated in arthritis (ADAMTS-1, ADAMTS-4 and ADAMTS-5) were cloned, and the sequences used to generate specific oligonucleotide primers for the RT-qPCR experiments. Our results show that genes encoding ADAM-10 and ADAM-17 were not induced in most laminitic animals, whereas ADAMTS-4 gene expression was strongly upregulated in nearly all horses with experimentally-induced and naturally acquired laminitis. The expression of matrix metalloproteases (MMP)-9 and ADAMTS 5 was also increased in many of the laminitic horses. In addition, TIMP-2 gene expression was decreased in most laminitic horses, whereas expression of genes encoding other TIMPs, namely TIMP-1 and TIMP-3, was randomly increased or decreased in the various models. We conclude that increased expression of lamellar ADAMTS-4 is a common feature of laminitis consistent with a central role of the gene product in the pathophysiology of the disease. PMID- 19131118 TI - Channel catfish, Ictalurus punctatus Rafinesque 1818, tetraspanin membrane protein family: characterization and expression analysis of CD81 cDNA. AB - CD81, also known as the target of an antiproliferative antibody 1 (TAPA-1) in human, is a member of tetraspanin integral membrane protein family. This protein plays many important roles in immune and other physiological functions. In this report, we characterized and analyzed expression of the channel catfish CD81 transcript. The full-length of channel catfish CD81 cDNA comprised of 1130 nucleotides, including an open reading frame which appears to encode a putative peptide of 234 amino acid residues. By comparison with the human counterpart, the channel catfish CD81 peptide could be divided into domains, including four transmembrane domains, three intracellular domains, and one of each small and large extracellular loops. The degree of conservation of the channel catfish CD81 amino acid sequence to that of mammalian counterparts ranged from 65% to 67%. The large extracellular domain shows the least conservation between fish and mammals. However, the characteristic Cys(159)-Cys(160)-Gly(161) motif and Cys(176/188) in this domain were conserved. The channel catfish CD81 transcript was detected by RT-PCR in spleen, head kidney, liver, intestine, skin and gill. This result provides important information for further elucidating CD81 functions in channel catfish. PMID- 19131119 TI - Sepsis-From human organ failure to laminar failure. AB - The horse with gram negative sepsis is known to be at particular risk of succumbing to laminitis. This review summarizes recent evidence indicating that similar pathologic events relating to inflammatory injury occur in laminar failure in laminitis as occur in organ injury/failure in human sepsis. The discussion also points out some important differences between the laminae and target organs in human sepsis that impact the clinical nature of the disease. PMID- 19131120 TI - Gramine-induced growth inhibition, oxidative damage and antioxidant responses in freshwater cyanobacterium Microcystis aeruginosa. AB - In recent years, the exploration and development of the effective methods of treatment and prevention to algal blooms, especially Microcystis aeruginosa blooms has been an important issue in the field of water environment protection. Allelochemicals (natural plant toxins) are considered promising sources of algicides to control algal blooms. The objective of this study is to determine the inhibitory effects and potential mechanisms of a well-known allelochemical gramine (N,N-dimethyl-3-amino-methylindole) on bloom-forming cyanobacterium M. aeruginosa. The results showed that this indole alkaloid effectively inhibited the growth of M. aeruginosa. The effective concentration causing a 50% inhibition at 3 d (EC(50, 3 d)) increased with the initial algal density (IAD) increasing. When IAD increased from 5x10(4) to 5x10(5)cellsmL(-1), the values of EC(50, 3 d) increased from 0.5 to 2.1mgL(-1). In the cells of M. aeruginosa, gramine caused an obvious increase in the level of reactive oxygen species (ROS). The lipid peroxidation product malondialdehyde (MDA) increased significantly in gramine treated cells. The effects of gramine on enzymatic and non-enzymatic antioxidants were in different manners. The activity of superoxide dismutase (SOD) was decreased after gramine exposure. The catalase (CAT) activity was increased after 4h but decreased from 60h. Both the contents and the regeneration rates of ascorbic acid (AsA) and reduced glutathione (GSH) were increased after 4h of exposure to gramine. However, only GSH content was still increased after 40h of exposure. These results suggested that the activation of antioxidants in M. aeruginosa played an important role to resist the stress from gramine at initial time, the inactivation of SOD is crucial to the growth inhibition of M. aeruginosa by gramine, and the phytotoxicity of gramine on M. aeruginosa may be due to oxidative damage via oxidation of ROS. PMID- 19131121 TI - Effect of nonylphenol on giant freshwater prawn (Macrobrachium rosenbergii) via oral treatment: toxicity and messenger RNA expression of hemocyte genes. AB - A previous in vitro study has indicated that two alkylphenol ethoxylates (APEs) could potentially damage hemocytes and influence cellular immunity of prawns (Macrobrachium rosenbergii). The aim of this study was to investigate the effects of nonylphenol (NP) on susceptibility to a pathogen and on the mRNA expression of hemocyte genes, including four immune-related genes. NP at different concentrations was fed continuously to prawn (M. rosenbergii) for 1, 3, 6, and 9 days. Challenging prawns with Lactobacillus garvieae resulted in 44-50%, 20-24% and 10-12% mortality were detected after prawns were fed with 100, 10 and 1microNP/prawn for 6 days, respectively. In comparison with control prawns fed with phosphate-buffered solution (PBS), the increase of mRNA levels of four immune-related genes, alpha-2 microglobulin (alpha-2m), antimicrobial peptides (amp), peroxinectin (pon), and prophenoloxidase (propo), was detected on days 1, 3 and 6 after feeding with 100microg/prawn; on day 9, only the mRNA level of amp of the NP-treated group was significantly increased, while that of the remaining groups was not different from that of the control. In addition, two other hemocyte genes were also studied, including a respiration-related gene, cytochrome oxidase subunit (cos), and an unknown gene, L12X3. The mRNA level of cos was elevated during the experimental period, but an increase of L12X3 expression was detected only on day 1 after treatment. Regarding sensitivity of these genes to NP, the results from NP-treated prawns on day 1 after treatment revealed (1) that mRNA expression of the six genes in the 100-microg-NP-treated group was significantly different from that of control group, (2) that the mRNA levels of three immune-related genes (amp, pon and propo) in 10-microg-treated group were significantly higher than that of control group, and (3) that a significant change of propo was detected in 1-microg-treated group. These results suggest that NP may enhance the immune response of prawns, but the effect created by a high concentration of NP may damage prawns, and then increase the susceptibility to pathogen. PMID- 19131122 TI - Effects of muscle strength and aerobic training on basal serum levels of IGF-1 and cortisol in elderly women. AB - The aim of this study was to compare the effects of muscle strength and aerobic training on the basal serum levels of IGF-1 and cortisol in elderly women. The subjects were divided in three groups as follows. 1. Strength training group (SG) submitted to the weight training called 1-repetition maximum test (1-RM, 75-85%). This group contained 12 subjects of mean age=66.08+/-3.37 years; and body mass index (BMI)=26.0+/-3.72kg/m(2). (2) Aerobic training group (AG) submitted to aquatic exercise; they were 13 subjects of the mean age=68.69+/-4.70 years; and BMI=29.19+/-2.96kg/m(2). (3) A control group (CG) of 10 subjects, of mean age=68.80+/-5.41 years; BMI=29.70+/-2.82kg/m2. The training periods were 12 weeks, Fasting blood was analyzed to measure IGF-1 and basal cortisol levels (by chemiluminescence method), both at the beginning and the end of the intervention. Student's t-test revealed increased IGF-1 in the SG (p<0.05) compared to the other two groups. Repeated-measure ANOVA showed also elevated IGF-1 (p<0.05) in the SG compared to the other groups (AG and CG). There were no differences in cortisol levels. In conclusion, high-intensity training caused changes in IGF-1. This suggests that strength training may provoke anabolic effects in elderly individuals. PMID- 19131123 TI - The relationship of age and leg strength in the step test with stipulated tempo in the elderly. AB - This study aimed to examine age differences in the step test with stipulated tempo, a test of dynamic balance ability in the elderly. One hundred eighty-nine healthy elderly people (96 men and 93 women) executed the step test for 20 s twice, with adjustments of the metronome (40 bpm, 60 bpm and 120 bpm). The knee extension strength of both legs was each measured twice. Evaluation parameters included the time difference between metronome sound and the time when each foot hit the ground, two-leg support time, and one-leg support time. The time difference and two-leg support time were significantly longer in the order of 40 bpm, 60 bpm, and 120 bpm tempos, and one-leg support time was shorter in the 40 bpm tempo than the 60 bpm and 120 bpm tempos. A significant age difference was found in the 40 bpm or 60 bpm tempo, and the former two parameters were longer in the older groups and the one-leg support time was shorter. Three evaluation parameters showed significant relationships with knee extension strength. In conclusion, because the step test with a slow tempo (40 bpm or 60 bpm) requires longer one-leg support time, it is effective for the evaluation of dynamic balance ability and leg strength of the elderly. PMID- 19131124 TI - Biopsychosocial differences between drivers and non-drivers over the age of 74. AB - The growing aging population will lead to an increased number of elderly drivers. There are few epidemiological studies that have approached this issue. Our aim was to compare the biopsychosocial characteristics of elderly drivers vs. elderly non-drivers. The method was a door-to-door population-based cross-sectional study of a sample consisting of inhabitants over 75 years of age in the Angles (Girona) Basic Healthcare Area. The study protocol included an extensive biopsychosocial assessment using standardized instruments. Descriptive and logistic regression analyses were performed to identify the variables associated with driving. A total of 875 inhabitants participated in the study (82%). The prevalence of drivers in this age group was 24.8% (95% confidence interval-CI=21.9-27.7). The variables associated with driving were age, sex, visual acuity, physical self concept and cognitive and functional capacity. Of the drivers, 45.8% (CI=38.9 52.7) were undergoing treatment with central nervous system drugs. The findings of this study suggest that age, gender, visual acuity, functional independence, preserved cognitive functioning and positive physical self-concept are correlated to the driving ability in elderly aged 75 years and over. Implications for driving practices related with these variables are discussed. PMID- 19131125 TI - Impact of socioeconomic status on guideline-recommended care for ST-elevation myocardial infarction in the United States. PMID- 19131126 TI - Restless legs syndrome (RLS) in anemic patients with congestive heart failure and chronic renal failure: lack of effect of anemia treatment. AB - OBJECTIVE: To assess the prevalence of Restless Legs Syndrome (RLS) in anemic patients with Congestive Heart Failure (CHF) and Chronic Renal Failure (CRF) and to evaluate the effect of anemia treatment on RLS. METHODS: 38 anemic CHF-CRF patients were treated with subcutaneous Erythropoietin (EPO) and intravenous (IV) iron over 1 year. They were questioned initially and at 3 months post treatment about symptoms of RLS according to standard criteria. They were also contacted by telephone about RLS symptoms 12 months after onset of anemia treatment. RESULTS: RLS was found in 15 (39.5%) of the 38 patients. In 10 (66.7%) patients it was present at least 6 days a week. The prevalence of the RLS initially was not related to Hb, to serum iron or % Transferrin Saturation. Diabetes and lower serum ferritin were more common in the RLS group (p<0.05). After 3 months of treatment, Hb increased from 10.4+/-0.8 to 12.3+/-1.2 g/dl, but RLS symptoms did not change. By 12 months the prevalence and frequency of RLS complaints was similar to what it had been initially. CONCLUSION: RLS is common and often undiagnosed and untreated in anemic CHF-CRF patients. Unfortunately, successful treatment of anemia with EPO and IV iron did not improve this condition. PMID- 19131128 TI - The treatment of clopidogrel resistance: triple antiplatelet therapy and future directions. PMID- 19131127 TI - Metabolic syndrome as a predictor of cardiovascular diseases and type 2 diabetes in Koreans. AB - BACKGROUND: The purpose of this study was to investigate whether the presence of the NCEP-ATP III defined metabolic syndrome (MS) is associated with the future development of cardiovascular disease (CVD) and diabetes in Koreans. METHODS: The study subjects were recruited from among those who visited the Health Promotion Center at the Samsung Medical Center. 2435 subjects (1761 men and 674 women), 20 to 78 years of age, were enrolled and evaluated for the development of new onset CVD (coronary heart disease [CHD] and stroke) during a mean 8.7 years of follow up. RESULTS: The prevalence of the MS at baseline was 21.7% (382/1761) and 11.4% (77/674) in men and women respectively, and the MS was found to be associated with the risk for CVD in both men and women (OR, 1.98; 95% CI, 1.30-3.03 in men, 4.04; 95% CI, 1.78-9.14 in women). More specifically, the MS was associated with the risk for future CHD (OR, 3.68; 95% CI, 1.93-7.01) in men and stroke (OR, 3.96; 95% CI, 1.58-9.94) in women. However, no statistical differences were found between the HOMA-IR tertiles with regard to the risk for CVD. After controlling for fasting plasma glucose (FPG) levels, the predictive power of the MS for an increased risk for diabetes was dramatically decreased (OR, from 3.69 to 1.77) in men, and it no longer was a predictor in women. CONCLUSIONS: The NCEP-defined MS was found to be associated with the risk for future CVD, i.e., CHD in men and stroke in women. PMID- 19131129 TI - Buerger's disease-like vasculitis associated with Kimura's disease. AB - A 46-year-old man was first diagnosed as Buerger's disease according to his clinical and radiological features because he had no evidence of parasitic, allergic and connective tissue disease. Soft subcutaneous nodules suspected of lymphadenopathy on the bilateral inguinal regions were recognized after admission. Positron emission tomography scan showed the increased uptake of (18)F fluoro-2-deoxyglucose in the bilateral inguinal regions. We finally diagnosed him as Kimura's disease based on pathologic findings and laboratory data, and started steroid therapy. The uptake of (18)F-fluoro-2-deoxyglucose disappeared and his leg pain was improved after the treatment. This is the first case report presenting a patient of Kimura's disease with Buerger's disease-like vasculitis who was demonstrated by positron emission tomography. PMID- 19131130 TI - HIV-associated vascular diseases: structural and functional changes, clinical implications. AB - After more than two decades of AIDS epidemic, the spectrum of HIV-associated vascular diseases has mainly evolved from infectious and inflammatory vasculitides to premature atherosclerosis, its related contributing conditions (metabolic syndrome, dyslipidemia, insulin resistance syndrome) and complications (acute coronary and cerebrovascular syndromes). Today, as the AIDS epidemic further progresses worldwide and as the life expectancy of HIV-infected patients treated with effective antiviral regimens has dramatically increased, more than 10% of patients experience cardiovascular manifestations. The complex interplay between viral infection, inflammatory and cytokines pathways, protease inhibitors induced hyperlipidemia and direct effects on endothelial cells has not, by far, been integrated in a single comprehensive pathogenesis network. However, recognition of its main components has resulted in a broader appreciation of cardiovascular risk and risk factors in HIV-infected/treated patients. Cardiovascular prevention is required in more than one half of HIV infected/treated patients to achieve a reliable effectiveness of modern antiretroviral therapy. As the prognosis of HIV patients improves continuously, this rate is also likely to increase in the future. PMID- 19131131 TI - Heart in the heart: dual faced primary cardiac lymphoma on PET-CT. AB - We describe a 71 year-old woman with primary cardiac lymphoma which was diagnosed by multimodality imaging particularly with PET/CT scan and finally confirmed by open biopsy. "Heart-shaped" bright signal in the right heart on PET-CT scan was another face of an aggressive cancer: primary cardiac lymphoma which is extremely rare and fatal. PMID- 19131132 TI - Timing of granulocyte-colony stimulating factor treatment after acute myocardial infarction and recovery of left ventricular function: results from the STEMMI trial. AB - BACKGROUND: Granulocyte-colony stimulating factor (G-CSF) therapy after ST elevation myocardial infarction (STEMI) have not demonstrated impact on systolic recovery compared to placebo. However, recent studies suggest that timing of G CSF therapy is crucial. METHODS: Timing of G-CSF treatment was analyzed in the STEMMI MRI subpopulation including 54 patients with STEMI treated with primary percutaneous coronary intervention (PCI) <12 h after symptom onset. Patients were randomized to double blind treatment with G-CSF (10 microg/kg/day) or placebo. Treatment was initiated from 17 to 65 h (mean 30) after PCI. Left ventricular ejection fraction (LVEF) was evaluated with MRI. RESULTS: Recovery of LVEF from baseline to 6 months was not associated with time from PCI to G-CSF. An identical improvement in LVEF was found in the placebo group and the G-CSF group (p=0.8). There was no correlation between time from PCI to G-CSF and maximum plasma concentration of CD34+ cells (r=-0.3, p=0.1). Similar results were found from data on recovery of the infarction size and change in the systolic wall thickening. CONCLUSIONS: In the time window from 17 to 65 h after STEMI treated with PCI, the timing of G-CSF treatment does not seem to affect the recovery of LVEF. It remains to be determined if very early, or very late G-CSF treatment might be effective. PMID- 19131133 TI - Revelation of an asymptomatic ventricular septal defect in elderly patient before a surgical intervention. AB - Certain congenital cardiac defects may go undetected for several years due to lack of symptoms and signs. The natural history of ventricular septal defects depends on the size of the defect and on the pulmonary resistance. In adults congenital heart disease, ventricular septal defects represent about 10% of the cases. However, the finding of a ventricular septal defect in an elderly individual >80 years of age is extremely uncommon and only two cases have been reported in a living patient, as an accidental finding. We describe a case of an asymptomatic ventricular septal defect associated with interventricular septal hypertrophy, aortic regurgitation, mitral stenosis and regurgitation in an 83 year-old Italian man before a surgical treatment for inguinal hernia repair. To our knowledge, this is the third report of a ventricular septal defect in a living elderly individual >80 years of age. PMID- 19131134 TI - Adult presentation of combined unilateral atresia of the right proximal pulmonary artery and left patent ductus arteriosus: case report and embryological considerations. AB - We report on a rare case of combined unilateral atresia of the proximal right pulmonary artery (PA) and left patent ductus arteriosus (PDA). A 46 year-old female patient with known PDA and associated advanced pulmonary arterial hypertension presented with progressive hemoptysis and increasing exertional dyspnea. Computed tomography of the chest proved the presence of the known PDA but surprisingly failed to demonstrate an extrapulmonary proximal right PA. We show the imaging features and discuss the embryological background of this rare congenital cardiovascular malformation. PMID- 19131135 TI - Incidence and determinants of myocardial infarction following percutaneous coronary interventions according to the revised Joint Task Force definition of troponin T elevation. AB - BACKGROUND: Elevations in troponin T (TnT) occur frequently following percutaneous coronary intervention (PCI) and are associated with an adverse prognosis. The Joint ESC/ACC/AHA/WHF Task Force have released a proposal for a universal definition of myocardial infarction (MI), including diagnostic criteria for PCI associated MI. This is based on a TnT cut-point of more than three times the 99th percentile (0.03 ng/ml), which better reflects the precision of the assay. Our study investigated the incidence and predictive factors of a PCI associated MI, using the revised definition. METHODS: 325 patients were studied following PCI with stenting. TnT was collected at both 8 and 18 h following PCI in patients with either stable or unstable angina and normal baseline TnT levels. Comparison was made of both clinical and procedural characteristics of patients with and without a rise in TnT following intervention, using cut points of 0.01 and 0.03 ng/ml. RESULTS: TnT was elevated > or = 0.03 ng/ml in 27% and > or = 0.01 ng/ml in 39% of patients following PCI. Troponin elevation was significantly more likely in those patients who experienced peri-procedural ischemic symptoms or EKG changes, or in whom abciximab was used. The variables associated with a troponin rise showed a greater difference between TnT positive and negative patients when using 0.03 ng/ml compared to 0.01 ng/ml, suggesting that this may be a better definition of PCI-related MI. CONCLUSIONS: Approximately one-quarter of low risk patients experience a procedural MI according to the revised definition. Rises in troponin were significantly associated with peri-procedural ischemic symptoms and EKG changes, and abciximab use, consistent with this level of TnT reflecting true myocardial necrosis. PMID- 19131136 TI - Bimanual coordination as task-dependent linear control policies. AB - When we perform actions with two hands in everyday life, coordination has to change very quickly depending on task goals. Here, we study these task-dependent changes using a bimanual reaching task in which participants move two separate cursors to two visual targets, or move a single cursor, displayed at the average position of the two hands, to a single target. During the movement, one of the hands is perturbed in a random direction using a viscous curl field. We have previously shown that feedback control, the structure of noise, and adaptation change between these two tasks as predicted by optimal control theory: feedback control is independent when the hands control two cursors, but becomes dependent when they move one cursor together. The same changes are observed even on trials in which no visual feedback about the cursor position is given. One assumption in this model is that coordinative motor commands can be described as a linear function of the state of the left and right hands. Here we test the assumption by studying the feedback corrections for 25 combinations of force fields applied to the two hands. Our study shows that feedback gains are constant across all levels of force fields strength, providing strong evidence that intermanual coordination for this task can accurately be explained by optimal task-dependent linear feedback gains. PMID- 19131137 TI - Evaluation of different procedures for the optimized detection of Vibrio parahaemolyticus in mussels and environmental samples. AB - Vibrio parahaemolyticus is a marine bacterium with a worldwide distribution and is frequently associated with human outbreaks of infection. Detection and isolation of V. parahaemolyticus from natural sources is often problematical because of limitations in the analytical procedures. We evaluated a combination of conventional and molecular protocols previously described for the investigation of V. parahaemolyticus, with the aim of identifying the best procedures for improved detection of this organism in environmental matrixes. A total of 259 samples of zooplankton (103), mussels (48) and seawater (108) were investigated by an Absence-Presence method (A/P), whereas 118 samples of zooplankton (70) and mussels (48) were analyzed by the Most Probable Number (MPN) method. All samples were processed by a two-step enrichment procedure, firstly with APW broth and then with SPB as selective secondary broth. Detection of V. parahaemolyticus was by direct-PCR and by plate culture on TCBS and CHROMagar Vibrio, after sample enrichment in APW and SPB. With the A/P method, V. parahaemolyticus was detected in 23.6% samples by direct-PCR, whereas only 11.2% samples were positive with the plate culture method. With the MPN method, V. parahaemolyticus was detected in 54.2% and 27.1% of the samples by direct-PCR and plate culture respectively; this indicated the existence of 31% false negative results with the A/P method. No significant differences between the use of a single (APW) or two-step enrichment (APW+SPB) were observed by direct-PCR with A/P or MPN, although a significant higher presence of V. parahaemolyticus was detected by plate culture in both protocols with the two-step enrichment procedure. In conclusion, direct-PCR after sample enrichment in APW broth was the most successful method for detection of V. parahaemolyticus with the A/P procedure and enumeration by MPN. Better detection was obtained with MPN than with the A/P protocol. Conversely, the plate culture procedure showed better results with the two-step enrichment protocol in which CHROMagar Vibrio was used as the selective agar. PMID- 19131138 TI - Estimated incidence of gestational diabetes mellitus in Hungary. PMID- 19131139 TI - Interactive computer program for optimal designs of longitudinal cohort studies. AB - Many large scale longitudinal cohort studies have been carried out or are ongoing in different fields of science. Such studies need a careful planning to obtain the desired quality of results with the available resources. In the past, a number of researches have been performed on optimal designs for longitudinal studies. However, there was no computer program yet available to help researchers to plan their longitudinal cohort design in an optimal way. A new interactive computer program for the optimization of designs of longitudinal cohort studies is therefore presented. The computer program helps users to identify the optimal cohort design with an optimal number of repeated measurements per subject and an optimal allocations of time points within a given study period. Further, users can compute the loss in relative efficiencies of any other alternative design compared to the optimal one. The computer program is described and illustrated using a practical example. PMID- 19131140 TI - Anti-lung cancer effects of novel ginsenoside 25-OCH(3)-PPD. AB - 20(S)-25-methoxyl-dammarane-3beta, 12beta, 20-triol (25-OCH(3)-PPD), a newly identified natural product from Panax notoginseng, exhibits activity against a variety of cancer cells. Herein, we report the effects of this compound on human A549, H358, and H838 lung cancer cells, and compare these effects with a control lung epithelial cell line, BEAS-2B. 25-OCH(3)-PPD decreased survival, inhibited proliferation, and induced apoptosis and G1 cell cycle arrest in the lung cancer cell lines. The P. notoginseng compound also decreased the levels of proteins associated with cell proliferation and cell survival. Moreover, 25-OCH(3)-PPD inhibited the growth of A549 lung cancer xenograft tumors. 25-OCH(3)-PPD demonstrated low toxicity to non-cancer cells, and no observable toxicity was seen when the compound was administered to animals. In conclusion, our preclinical data indicate that 25-OCH(3)-PPD is a potential therapeutic agent in vitro and in vivo, and further preclinical and clinical development of this agent for lung cancer is warranted. PMID- 19131141 TI - ER-associated complexes (ERACs) containing aggregated cystic fibrosis transmembrane conductance regulator (CFTR) are degraded by autophagy. AB - The ubiquitin-proteasome pathway and autophagy are the two major mechanisms responsible for the clearance of cellular proteins. We have used the yeast Saccharomyces cerevisiae as a model system and the cystic fibrosis transmembrane conductance regulator (CFTR) as a model substrate to study the interactive function of these two pathways in the degradation of misfolded proteins. EGFP tagged human CFTR was introduced into yeast and expressed under a copper inducible promoter. The localization and degradation of EGFP-CFTR in live cells were monitored by time-lapse imaging following its de novo synthesis. EGFP-CFTR first appears within the perinuclear and sub-cortical ER and is mobile within the plane of the membrane as assessed by fluorescence recovery after photobleaching (FRAP). This pool of EGFP-CFTR is subsequently degraded through a proteasome dependent pathway that is inhibited in the pre1-1 yeast strain defective in proteasomal degradation. Prolonged expression of EGFP-CFTR leads to the sequestration of EGFP-CFTR molecules into ER structures called ER-associated complexes (ERACs). The sequestration of EGFP-CFTR into ERACs appears to be driven by aggregation since EGFP-CFTR molecules present within ERACs are immobile as measured by FRAP. Individual ERACs are cleared from cells through the autophagic pathway that is blocked in the atg6Delta and atg1Delta yeast strains defective in autophagy. Our results suggest that the proteasomal and the autophagic pathways function together to clear misfolded proteins from the ER. PMID- 19131142 TI - Super-elevated CO2 interferes with stomatal response to ABA and night closure in soybean (Glycine max). AB - Studies have shown stomatal conductance (g(s)) of plants exposed to super elevated CO2 (>5000micromol mol(-1)) increases in several species, in contrast to a decrease of g(s) caused by moderate CO2 enrichment. We conducted a series of experiments to determine whether super-elevated CO2 alters stomatal development and/or interferes with stomatal closure in soybean (Glycine max). Plants were grown at nominal ambient (400), elevated (1200) and super-elevated (10,000micromol mol(-1)) CO2 in controlled environmental chambers. Stomatal density of the plant leaf was examined by a scanning electron microscope (SEM), while the stomatal response to the application of exogenous abscisic acid (ABA), a phytohormone associated with water stress and stomatal control, was investigated in intact growing plants by measuring the g(s) of abaxial leaf surfaces using a steady-state porometer. Relative to the control (400micromol mol(-1) CO2) plants, daytime stomatal conductance (g(s,day)) of the plants grown under 1200 and 10,000micromol mol(-1) CO2 was reduced by 38% and 15%, respectively. Dark period stomatal conductance (g(s,night)) was unaffected by growing under 1200mumol mol(-1) CO2) but dramatically increased under 10,000micromol mol(-1) CO2. Stomatal density increased by 10% in the leaves of 10,000micromol mol(-1) CO2-grown plants, which in part contributed to the higher g(s,night) values. Elevating [CO2] to 1200micromol mol(-1) enhanced ABA-induced stomatal closure, but further increasing CO2 to 10,000micromol mol(-1) significantly reduced ABA-induced stomatal closure. These results demonstrated that stomatal response to ABA is CO2 dependent. Hence, a stomatal failure to effectively respond to an ABA signal and to close at night under extremely high CO2 may increase plants susceptibility to other abiotic stresses. PMID- 19131143 TI - Homocysteine promotes proliferation and activation of microglia. AB - Epidemiological and experimental studies have correlated hyperhomocysteinemia to a range of neurodegenerative conditions, including Alzheimer's disease, stroke, and Parkinson's disease. Although homocysteine-induced apoptosis in neurons has been extensively studied, little information is available regarding the effect of homocysteine on microglia. In this report, we demonstrated that homocysteine promoted proliferation and up-regulated the expression of CD11b (a marker of microglial activation). Consistent with our in vitro results, a significant increase in the number of CD11b-positive microglia was also observed in brain sections of mice with hyperhomocysteinemia. Homocysteine promoted the activity of NAD(P)H oxidases, resulting in the generation of reactive oxygen species. Up regulation of NAD(P)H oxidase activity by homocysteine appears to be due to its ability to induce the phosphorylation of p47phox through the p38 MAPK pathway. Furthermore, inhibition of reactive oxygen species significantly blocked cellular proliferation and activation in microglia. Since microglial proliferation and activation play an important role in the development of several neurodegenerative disorders, our results reveal a novel role of homocysteine in the pathogenesis of neurodegenerative diseases. PMID- 19131144 TI - [Current profile of peritoneal tuberculosis: study of a Tunisian series of 42 cases and review of the literature]. AB - INTRODUCTION: The aim of this study was to determine the epidemiological, clinical and therapeutic features, and the outcome of peritoneal tuberculosis in an endemic area of tuberculosis on the basis of our experience. METHODS: All cases of peritoneal tuberculosis confirmed by histologic examination and hospitalized in the department of gastroenterology of Hedi-Chaker hospital between January 1987 and December 2006 were analyzed retrospectively. RESULTS: Forty-two cases (mean age 38 years) were included in this study. Clinical presentation was dominated by ascites (100%), fever (76.2%) and abdominal pain (73.8%). The average delay for consultation after the first symptom was 3.6 months. The ascites was exudative in 100% of cases with lymphocytic predominance in 96.6%. The tuberculous skin test was above 10mm in 39.1% of cases. All of our patients had a peritoneal exploration. A nodular aspect of the peritoneal cavity was found in 40 patients (95.2%). Adhesions were noted in 25 patients. Therapeutic protocols varied among years. The outcome was favorable in all patients. CONCLUSION: Diagnosing peritoneal tuberculosis is a challenge for clinicians. Coelioscopy with peritoneal biopsies still remains the method of choice to establish a definite diagnosis of peritoneal tuberculosis. PMID- 19131145 TI - [Liver infarcts with early HELLP syndrome and probable catastrophic antiphospholipid syndrome]. AB - INTRODUCTION: Among microangiopathic disorders of pregnancy, catastrophic antiphospholipid syndrome (CAPS) is a maternal and fetal life-threatening disorder. Hepatic involvement of this multi-systemic disorder can be confused with HELLP syndrome, occurring usually later in the course of pregnancy. CASE REPORT: We report a case of probable CAPS with hepatic disease in a pregnant woman at 13 week's gestation, with antiphospholipid syndrome and biological features of HELLP syndrome. Unspecific hepatic imaging, well-described in our case allowed undelayed therapy. CONCLUSION: CAPS and HELLP syndrome, both severe microangiopathic disorders, may be associated. Nosological distinction does not modify treatment strategy, which is a maternal and foetal emergency, but their overlapping requires aggressive and early management. PMID- 19131146 TI - Online multimedia delivery of the European Union directive within adult nursing education. AB - The use of online multimedia content in nurse education is still in its infancy with limited research about its role and acceptance by students and staff within nursing courses. This paper describes the development of interactive web-based delivery for the European Union directive (77/453/EEC) for maternity, child care and mental health within adult nursing education. An evaluative study carried out by an online questionnaire with students, using both structured questions and free text, and a focus group with staff whilst identifying areas for further development found general acceptance of this approach. The study has provided important insights into both the content and delivery of the packs, and the format of the material. The use of IT, with multimedia elements, was seen as being beneficial by both students and staff. The findings will be useful in improving and developing the packs for future cohorts. PMID- 19131147 TI - Two adenomatoid odontogenic tumours of the maxilla: a case report. AB - An adenomatoid odontogenic tumour (AOT) is a well-recognised benign tumour that enlarges slowly. We report here the rare occurrence of two separate tumours in the anterior maxillary region that have not been documented previously. They were associated with unusual findings such as relatively large size, a supernumerary tooth, rapid growth with cortical perforation, and multilocular appearance with root resorption on plain radiograph. PMID- 19131148 TI - The Leeds method for titanium cranioplasty construction. PMID- 19131149 TI - Comparison of sleep/wake behavior in CKD stages 4 to 5 and hemodialysis populations using wrist actigraphy. AB - BACKGROUND: Patients with kidney failure have more sleep symptoms than the general population, but the contribution to sleep symptoms of kidney failure versus its treatment with thrice-weekly hemodialysis has been unclear. We assessed the influence of hemodialysis on sleep/wake behavior by using wrist actigraphy and self-reported sleep quality compared with patients with chronic kidney disease (CKD) stages 4 to 5. STUDY DESIGN: Cross-sectional study. SETTING & PARTICIPANTS: Thirty-six patients with CKD stages 4 to 5 and 51 hemodialysis (HD) patients wore wrist actigraphs and completed sleep diaries for 2 weeks. PREDICTORS: Thrice-weekly HD versus CKD stages 4 to 5, unstable total sleep times (TSTs), early HD shift. OUTCOMES: Self-reported sleep quality and objective measures of sleep/wake behavior. MEASUREMENTS: Diaries, sleep questionnaires, and wrist actigraphy were performed. RESULTS: The group with CKD stages 4 to 5 had an average age of 51 years, 69% were men, 19% were African American, and average body mass index was 28.9 kg/m2. The HD group had an average age of 54 years, 60.8% were men, 49% were African American, and average body mass index was 27.5 kg/m2. Average TST was 66.8 minutes shorter and sleep efficiency was 5.2% lower in the HD group compared with the population with CKD stages 4 to 5. In the HD population, 28 individuals had a mean change in TST greater than 60 minutes between HD and non-HD nights, and this unstable sleep pattern was associated with daytime sleepiness. The early-HD group had TST 62 minutes (95% confidence interval, approximately 22 to 102) shorter than those with later HD shifts. No significant differences in sleep efficiency or fragmentation index were found between the early- and late-HD groups. LIMITATIONS: Study included those older than 18 years. CONCLUSIONS: Both patients with CKD stages 4 to 5 and HD patients have short and fragmented sleep. An early-morning HD shift was associated with shorter TST and greater variation in nightly TST. Additional trials of the possible beneficial effect of behavioral sleep interventions, more frequent HD, and later HD shifts on sleep patterns are needed. PMID- 19131150 TI - A meta-analysis of behavioral treatments for attention-deficit/hyperactivity disorder. AB - There is currently controversy regarding the need for and the effectiveness of behavior modification for children with attention-deficit hyperactivity disorder (ADHD) despite years of study and multiple investigations reporting beneficial effects of the intervention. A meta-analysis was conducted by identifying relevant behavioral treatment studies in the literature. One-hundred seventy-four studies of behavioral treatment were identified from 114 individual papers that were appropriate for the meta-analysis. Effect sizes varied by study design but not generally by other study characteristics, such as the demographic variables of the participants in the studies. Overall unweighted effect sizes in between group studies (.83), pre-post studies (.70), within group studies (2.64), and single subject studies (3.78) indicated that behavioral treatments are highly effective. Based on these results, there is strong and consistent evidence that behavioral treatments are effective for treating ADHD. PMID- 19131151 TI - Preventive effect of an iron varnish on bovine enamel erosion in vitro. AB - OBJECTIVE: The aim of this study was to evaluate, in vitro, the effect of an experimental varnish containing iron on the dissolution of bovine enamel by carbonated beverage. METHODS: Eighty specimens were randomly allocated to four groups (n=20 per group), according to the following treatments: Fe varnish (FeV, 10 mmol/L Fe), F varnish (FV, 2.71% F), placebo varnish (PV) and control (not treated, NT). The varnishes were applied in a thin layer and removed after 6h. Then, the samples were submitted to six cycles, alternating re- and demineralisation (only 1 day). Demineralisation was performed with the beverage Coca-Cola (10 min, 30 mL/block) and remineralisation with artificial saliva for 1h. In order to determine the amount of enamel dissolved, the wear was analysed by profilometry. Data were analysed by ANOVA and Tukey's test (p<0.05). RESULTS: The mean wear (+/-S.E.) was significantly lesser for the FeV (0.451+/-0.018 microm) when compared to the other treatments. The FV caused significantly less wear (0.554+/-0.022 microm) when compared to PV (0.991+/-0.039 microm) and NT (1.014+/-0.033), which did not significantly differ from each other. CONCLUSIONS: The results suggest that the iron varnish can interfere with the dissolution of dental enamel in the presence of acidic beverages. PMID- 19131152 TI - Effect of sodium sulfinate salts on the polymerization characteristics of dual cured resin cement systems exposed to attenuated light-activation. AB - OBJECTIVES: To evaluate the effectiveness of sodium sulfinate salt co-initiators on the degree of conversion (DC) of dual-cured cementing systems [combination of bonding agent (BA) and resin cement (RC)] when the activating light is attenuated or not available. METHODS: Two 5th generation products [Prime&Bond NT Dual Cure/Calibra (Dentsply), and Excite DSC/Variolink II (Ivoclar/Vivadent)] were applied, either with or without co-initiators (sodium sulfinate salts) added to the bonding agents, to the surface of a horizontal attenuated-total-reflectance unit, and were polymerized using one of four conditions: self-cure (SC), or direct light exposure (DLE/XL3000/3M) through a glass slide or through pre-cured resin discs (shades A2 or A4/2mm-thick) (Z250/3M-ESPE). Real-time infrared spectra were obtained after placing components (uncured) and during 10 min (cured) (16 scans at 2 cm(-1), FTS-40/Digilab/Bio-Rad) to obtain polymerization kinetics. DC and maximum rate of polymerization (Rpmax) were calculated using standard techniques of observing changes in aliphatic-to-aromatic peak ratios pre and post-curing. Data (n=5) were analyzed by two-way ANOVA and Tukey's test (p=0.05). RESULTS: The addition of co-initiators increased the DC in most of the groups. No significant difference in DC was observed between Prime Bond NT Dual Cure/Calibra and its version without co-initiator using the DLE curing mode. All DLE groups exhibited the highest DC after 10 min. No significant difference in DC was observed between A2 and A4 pre-cured overlay groups. The SC groups exhibited lower DC than the light-activated groups regardless of the presence of co initiator. Use of co-initiators increased the Rpmax only in the SC groups. The A2 and A4 groups exhibited lower Rpmax than DLE groups, regardless of the presence of co-initiator. CONCLUSION: The detrimental effects of light attenuation on the DC were not avoided by inclusion of sodium sulfinate salts, despite their ability to eliminate the chemical incompatibility between acidic monomers and self-curing components. PMID- 19131153 TI - A prospective randomized clinical trial of one bis-GMA-based and two ormocer based composite restorative systems in class II cavities: Five-year results. AB - OBJECTIVES: Ormocer composites, consisting of a silicon-based polymer, have been developed recently as a tooth-colored restorative material. The purpose of this prospective randomized clinical trial was to evaluate the performance of two small-particle hybrid ormocer-based restorative systems (AD, Admira/Admira Bond, VOCO; DE, Definite/Etch & Prime 3.0, Dentsply) and one small-particle hybrid bis GMA-based composite restorative system (TC, Tetric-Ceram/Syntac, Ivoclar Vivadent) in class II cavities. METHODS: From 128 occlusal-proximal restorations (44 AD, 43 DE and 41 TC) placed in 32 adult patients, eventually 77 (22 AD, 29 DE and 26 TC) remained available for evaluation after 5 years. Their clinical performance was scored according to the USPHS criteria and evaluation of bite wing radiographs. RESULTS: After 5 years, eight AD, six DE and seven TC restorations had failed (p=0.10, log-rank test). The main reason was fracture or marginal gap formation, while secondary caries accounted for four failures. In all restorations the quality of surface, margins and contact point decreased significantly compared to baseline. DE had a significant poorer color match (p<0.01). Statistical evaluation using the KW test showed that failures were concentrated on specific patients. CONCLUSIONS: In a group of class II restorations, there was no significant difference in failures after 5 years between ormocer-based and bis-GMA-based restorative systems. PMID- 19131154 TI - Multipathogenetic origin of a pelvic mass. AB - A 69-yr-old woman presented with a bulky hypogastric mass and abdominal pain. Computed tomography scan showed a mass anterosuperior and contiguous to the bladder wall, with a hypodense content, a voluminous bladder stone, and bilateral hydroureteronephrosis. Intraoperatively, the supravesical mass had the appearance of an infected urachal cyst. An unsuspected high-grade noninvasive papillary transitional cell carcinoma (TCC) of the bladder thoroughly surrounding the bladder stone became evident during the cystolithotomy. Postoperative videourodynamic study showed a normal voiding pattern with bilateral grade 4 vesicoureteral reflux. Early cystectomy was performed for uncontrolled recurrent bladder cancer, and the final pathology indicated pT1G3N0 TCC. PMID- 19131155 TI - The relation of testosterone levels with fatigue and apathy in Parkinson's disease. AB - OBJECTIVE: Fatigue and apathy are frequent in patients with Parkinson's disease (PD). Testosterone deficiency in male patients may contribute in development of fatigue and apathy as well. We investigated whether a possible relation exists between serum testosterone levels, fatigue and apathy in male PD patients. MATERIALS AND METHODS: We included 29 non-demented and non-depressed PD patients and 30 age- and sex-matched healthy subjects. Fatigue Severity Scale (FSS) and Apathy Evaluation Scale (AES-C) were used for the evaluations. In PD patients and healthy subjects, a relationship between FSS, AES-C scores and plasma testosterone levels were assessed. In addition, a correlation between FSS, AES-C and Unified Parkinson's Disease Rating Scale was investigated in PD group. RESULTS: The mean scores of FSS and AES-C were significantly higher in PD patients than those of the control group. The Unified Parkinson's Disease Rating Scale (UPDRS) scores were significantly correlated with FSS and AES-C scores. Mean free testosterone level was significantly lower in PD patients than controls (p=0.008). f-Testosterone levels of PD patients were not correlated with FSS or AES-C scores. CONCLUSION: Apathy and fatigue are frequent in PD and show significant correlation with the severity of the disease. f-Testosterone levels are not related with apathy or fatigue in male PD patients and the role of testosterone in the pathophysiology of these non-motor symptoms is still controversial. PMID- 19131156 TI - S1P/S1P1 signaling stimulates cell migration and invasion in Wilms tumor. AB - Sphingosine-1-phosphate (S1P) is an important regulator of cellular functions via interaction with its receptors S1P(1-5). To date, nothing is known about the S1P receptor expression and the effects of S1P signaling in Wilms tumor. In this study, we found ubiquitous expression of S1P receptors in Wilms tumor specimens and cell lines. We demonstrated that S1P(1) acted as a promigratory modulator by employing S1P(1) antagonist VPC44116, S1P(1) siRNA and adenoviral transduction in Wilms tumor cells. Further, we clarified that S1P(1)-mediated migration occurred via Gi coupling and activation of PI3K and Rac1. In addition, S1P stimulated WiT49 cell invasion through S1P(1)/Gi signaling pathway. We consider that targeting S1P(1) may be a point of therapeutic intervention in Wilms tumor. PMID- 19131157 TI - Visible light photoelectrocatalysis with salicylic acid-modified TiO2 nanotube array electrode for p-nitrophenol degradation. AB - This research focused on immersion method synthesis of visible light active salicylic acid (SA)-modified TiO2 nanotube array electrode and its photoelectrocatalytic (PEC) activity. The SA-modified TiO2 nanotube array electrode was synthesized by immersing in SA solution with an anodized TiO2 nanotube array electrode. Scanning electron microscope (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), infrared spectroscopy (IR), UV-vis diffuse reflectance spectrum (DRS), and Surface photovoltage (SPV) were used to characterize this electrode. It was found that SA-modified TiO2 nanotube array electrode absorbed well into visible region and exhibited enhanced visible light PEC activity on the degradation of p-nitrophenol (PNP). The degradation efficiencies increased from 63 to 100% under UV light, and 79-100% under visible light (lambda>400 nm), compared with TiO2 nanotube array electrode. The enhanced PEC activity of SA-modified TiO2 nanotube array electrode was attributed to the amount of surface hydroxyl groups introduced by SA-modification and the extension of absorption wavelength range. PMID- 19131158 TI - Montmorillonite surface properties and sorption characteristics for heavy metal removal from aqueous solutions. AB - Surface properties of montmorillonite (MMT) and its adsorption characteristics for heavy metals have been investigated with nickel and copper as sorbate from aqueous solutions. Employing the potentiometric and mass titration techniques in batch experimental methods, the point of zero charge (PZC) and point of zero net proton charge (PZNPC) of MMT edges at different ionic strengths present pH(PZC) and pH(PZNPC) to be 3.4+/-0.2. A crossing point was observed for the proton adsorption vs. pH curves at different ionic strengths of KCl electrolyte and in investigating MMT remediation potentialities as sorbent for heavy metals polluted waters, the effects of heavy metal concentration, pH, MMT dosage, reaction time and temperature for Cu(2+) and Ni(2+) uptake were studied. The sorption of metal ions by MMT was pH dependent and the adsorption kinetics revealed sorption rate could be well fitted by the pseudo-second-order rate model. The data according to mass transfer and intraparticle diffusion models confirmed diffusion of solutes inside the clay particles as the rate-controlling step and more important for the adsorption rate than the external mass transfer. Adsorption isotherms showed that the uptake of Cu(2+) and Ni(2+) could be described by the Langmuir model and from calculations on thermodynamic parameters, the positive Delta G degrees values at different temperatures suggest that the sorption of both metal ions were non spontaneous. Change in enthalpy (Delta H degrees) for Ni(2+) and Cu(2+) were 28.9 and 13.27 kJ/mol K respectively, hence an endothermic diffusion process, as ion uptake increased with increase in temperature. Values of DeltaS degrees indicate low randomness at the solid/solution interface during the uptake of both Cu(2+) and Ni(2+) by MMT. Montmorillonite has a considerable potential for the removal of heavy metal cationic species from aqueous solution and wastewater. PMID- 19131159 TI - Adsorption of cadmium from aqueous solutions on sulfurized activated carbon prepared from nut shells. AB - Low-cost activated carbon, derived from nut shells, and its modified sample have been used as replacements for the current expensive methods of removing cadmium from aqueous solutions and waste waters. Adsorption of cadmium onto four kinds of activated carbons has been studied; prepared activated carbon (PAC), commercial activated carbon (CAC), and the sulfurized ones (SPAC & SCAC). The activated carbon has been derived, characterized, treated with sulfur and then utilized for the removal of Cd(2+). Sulfurizing agent (SO(2) gas) was successfully used in adsorbents' modification process at the ambient temperature. Samples were then characterized and tested as adsorbents of cadmium. Effect of some parameters such as contact time, initial concentration and pH were examined. With increasing pH, the adsorption of cadmium ions was increased and maximum removal, 92.4% for SPAC, was observed in pH>8.0 (C(0)=100mg/L). The H-type adsorption isotherms, obtained for the adsorbents, indicated a favorable process. Adsorption data on both prepared and commercial activated carbon, before and after sulfurization, followed both the Frendlich and Langmuir models. They were better fitted by Frendlich isotherm as compared to Langmuir. The maximum adsorption capacities were 90.09, 104.17, 126.58 and 142.86 mg/g for CAC, PAC, SCAC and SPAC, respectively. Accordingly, surface modification of activated carbons using SO(2) greatly enhanced cadmium removal. The reversibility of the process has been studied in a qualitative manner and it shows that the spent SPAC can be effectively regenerated for further use easily. PMID- 19131160 TI - Study on the adsorption of cadmium(II) from aqueous solution by D152 resin. AB - The feasibility of using D152 resin as an adsorbent for cadmium(II) was examined. The adsorption capacity of D152 resin for cadmium(II) ions was studied as a function of solution pH, temperature and contact time. The optimal pH for the adsorption of cadmium(II) ions was found to be 5.95 in the HAc-NaAc system. The maximum uptake capacity of cadmium(II) was estimated to 378 mg/g D152 resin at 298 K, at an initial pH value of 5.95. The results obtained from equilibrium adsorption studies were conformed to the Langmuir and Freundlich and the correlation coefficients have been evaluated. The apparent adsorption rate constant was k(298 K)=4.01 x 10(-5)s(-1), and the apparent activation energy was 2.78 kJ/mol. Thermodynamic parameters, DeltaS was 108 J/(mol K), heat of adsorption (Delta H) value of 12.8 kJ/mol indicated the endothermic nature of the adsorption process, and a decrease of Gibbs free energy (Delta G) with increasing temperature also indicated the spontaneous nature of the process, respectively. The elution tests were carried out using various mixed reagents. The maximum elution percent of cadmium(II) ions were obtained when the reagent 0.5 mol/L HCl was used. PMID- 19131161 TI - Biosorption of arsenic from aqueous solution using agricultural residue 'rice polish'. AB - 'Rice polish' (an agricultural residue) was utilized successfully for the removal of arsenic from aqueous solution. Various parameters viz. pH, biosorbent dosage, initial metal ion concentration and temperature were studied. Langmuir, Freundlich and Dubinin-Radushkevich (D-R) isotherm models were used and the system followed all three isotherms, showing sorption to be monolayer on the heterogeneous surface of the biosorbent. The maximum sorption capacity calculated using Langmuir model was 138.88 microg/g for As(III) at 20 degrees C and pH 7.0 and 147.05 microg/g at 20 degrees C and pH 4.0 for As(V). The mean sorption energy (E) calculated from D-R model indicated chemisorption nature of sorption. Study of thermodynamic parameters revealed the exothermic, spontaneous and feasible nature of sorption process in case of both As(III) and As(V). The pseudo second-order rate equation described better the kinetics of arsenic sorption with good correlation coefficients than pseudo-first-order equation. Mass transfer, intraparticle diffusion, richenberg and elovich models were applied to the data and it was found that initially the sorption of arsenic was governed by film diffusion followed by intraparticle diffusion. Rice polish was found to be efficient in removing arsenic from aqueous solution as compared to other biosorbents already used for the removal of arsenic. PMID- 19131162 TI - Assessing hazardous risks of human exposure to temple airborne polycyclic aromatic hydrocarbons. AB - We proposed an integrated probabilistic risk assessment framework based on reported data to quantify human health risks of temple goers/workers to airborne polycyclic aromatic hydrocarbons (PAHs) from incense burning in typical Taiwanese temples. The framework probabilistically integrates exposure, human respiratory tract, and incremental lifetime cancer risk (ILCR) models to quantitatively estimate size-dependent PAHs exposure in human lung regions and cancer risks for temple goers (moderate and high exposures) and temple workers (extreme exposure). Our results show that the ILCRs are greater than the acceptable level of 10(-6) for extreme and high exposure groups through inhalation route. The result also indicates that the higher ILCRs (10(-6) to 10(-4)) are found in ingestion and dermal contact routes for temple goers/workers. For personal extreme exposure to carcinogenic PAH in the temple, 95% probability total ILCR (TILCR) (9.87 x 10(-4) to 1.13 x 10(-3)) is much greater than the range of 10(-6) to 10(-4), indicating high potential health risk to temple workers. For temple goers with high and moderate exposure groups, however, the 95% probability TILCRs were estimated from 6.44 x 10(-5) to 7.50 x 10(-5) and 5.75 x 10(-6) to 6.99 x 10(-6), respectively. This study successfully offers a scientific basis for risk analysis due to incense burning to enhance broad risk management strategies for temple indoor air quality. PMID- 19131163 TI - Gamma-ray induced degradation of diazinon and atrazine in natural groundwaters. AB - Degradation of diazinon and atrazine pesticides present in natural groundwaters was investigated on a laboratory scale upon gamma-irradiation from a (60)Co source. The effects of pesticide type, initial concentration, characteristics of natural groundwater, potential radical scavengers and absorbed dose on efficiency of pesticide degradation were investigated using GC-MS. gamma-Irradiation experiments were carried out for three concentrations (i.e. 0.329, 1.643 and 3.286 microM/diazinon and 0.464, 2.318 and 4.636 microM/atrazine) with irradiation doses over the range 0.5-5.6 kGy for diazinon and 0.2-21 kGy for atrazine. gamma-Radiolysis showed that diazinon was much easier to degrade by ionizing radiation compared to atrazine in all natural groundwater samples. This was observed at the three initial concentrations over the range irradiation doses. The irradiation doses required for degradation of 50 and 90% diazinon (distilled water) and atrazine (humic aqueous solution) at the three concentrations were not sufficient to degrade the same concentrations in different natural groundwater samples. Moreover, the presence of naturally occurring inorganic scavengers in solutions of diazinon and atrazine decreased significantly the efficiency of radiolytic degradation of pesticides, especially at higher concentrations. PMID- 19131164 TI - Effects of alcohol-partitioning type and airflow on cosolvent flooding to benzene LNAPL saturated porous media. AB - This study fundamentally investigated the swelling and distribution of benzene light nonaqueous phase liquid (LNAPL) in porous media while cosolvent was flushed to the benzene-partially saturated system. Furthermore, the effects of simultaneous injection of cosolvent and air on the LNAPL behavior were visualized and thus quantified within a two-dimensional transparent porous medium. Partitioning types of alcohols affected dissolution of benzene entrapped in porous media. Tert-butanol (TBA) and 1-propanol floods apparently increased the LNAPL area, while a 70% ethanol flood reduced the LNAPL area by dissolution. Airflow facilitates mobilization of the swollen LNAPL by TBA and 1-propanol, while it facilitates dissolution of non-swollen LNAPL by ethanol. Therefore, LNAPL behavior during cosolvent flooding would be determined by partitioning type of alcohols and the presence of airflow. PMID- 19131165 TI - Use of rice straw as biosorbent for removal of Cu(II), Zn(II), Cd(II) and Hg(II) ions in industrial effluents. AB - Adsorption experiments were carried out using waste rice straw of several kinds as a biosorbent to adsorb Cu(II), Zn(II), Cd(II) and Hg(II) ions from aqueous solutions at room temperature. To achieve the best adsorption conditions the influence of pH and contact time were investigated. The isotherms of adsorption were fitted to the Freundlich equation. Based on the experimental data and Freundlich model, the adsorption order was Cd(II)>Cu(II)>Zn(II)>Hg(II) on the rice straw. This quick adsorption process reached the equilibrium before 1.5h, with maximum adsorptions at pH 5.0. Thermodynamic aspects of the adsorption process were investigated. The biosorbent material was used in columns for the removal of ions Cu, Zn, Cd and Hg of real samples of industrial effluent and its efficiency was studied. PMID- 19131166 TI - Examination of the photograph series of daily activities (PHODA) scale in chronic low back pain patients with high and low kinesiophobia. AB - The current study examined the Photograph Series of Daily Activities (PHODA) Scale in a sample of chronic low back pain patients with high and low levels of kinesiophobia (pain-related fear). Thirty-three participants completed a modified version of the PHODA (PHODA-M) that obtained ratings of both anticipated pain and harm. Participants' responses on the PHODA-M were compared to predicted and experienced pain and harm ratings collected during a graded-difficulty reaching task. Individuals with high kinesiophobia reported higher pain and harm expectancies for PHODA-M stimuli, as well as greater predicted and experienced pain and harm in response to movement performance. In comparison to low fear participants, high fear participants showed larger correlations between pain expectancies for movements depicted in the PHODA and their ratings of predicted and experienced pain on each movement comprising the reaching task. Additionally, high fear participants showed larger associations between the perceived harmfulness of PHODA activities and predicted and experienced harm ratings in response to the reaching task. Implications for the findings are discussed. PMID- 19131167 TI - APT analyses of deuterium-loaded Fe/V multi-layered films. AB - Interaction of hydrogen with metallic multi-layered thin films remains as a hot topic in recent days. Detailed knowledge on such chemically modulated systems is required if they are desired for application in hydrogen energy system as storage media. In this study, the deuterium concentration profile of Fe/V multi-layer was investigated by atom probe tomography (APT) at 60 and 30K. It is firstly shown that deuterium-loaded sample can easily react with oxygen at the Pd capping layer on Fe/V and therefore, it is highly desired to avoid any oxygen exposure after D(2) loading before APT analysis. The analysis temperature also has an impact on D concentration profile. The result taken at 60K shows clear traces of surface segregation of D atoms towards analysis surface. The observed diffusion profile of D allows us to estimate an apparent diffusion coefficient D. The calculated D at 60K is in the order of 10(-17)cm(2)/s, deviating 6 orders of magnitude from an extrapolated value. This was interpreted with alloying, D-trapping at defects and effects of the large extension to which the extrapolation was done. A D concentration profile taken at 30K shows no segregation anymore and a homogeneous distribution at c(D)=0.05(2)D/Me, which is in good accordance with that measured in the corresponding pressure-composition isotherm. PMID- 19131168 TI - A quantitative assessment of microelectrodes. AB - In order to improve the performance and applicability of atom probe tomography, the application of microelectrodes has been suggested and is realized in modern commercial instruments. In contrast to the original proposition by Nishikawa, in practical realization the down scaling of the microelectrode is limited to about 10microm due to the requirements of a stable measurement. In this work, the field enhancement by electrodes of this size was measured in the FIM mode and compared to finite element calculations of the electric field. The experimental data reveal considerable scattering between individual microelectrodes and specimen tips, but on the average the predictions by the finite element calculation are confirmed. Even a microelectrode of 50microm diameter yields a reasonable field enhancement close to a factor of two. PMID- 19131169 TI - Amplitude and frequency modulation torsional resonance mode atomic force microscopy of a mineral surface. AB - Scanning probe imaging in a shear force mode allows for the characterization of in-plane surface properties. In a standard AFM, shear force imaging can be realized by the torsional resonance mode. In order to investigate the imaging conditions on mineral surfaces, a torsional resonance mode atomic force microscope was operated in amplitude (AM) and frequency modulation (FM) feedback. Freshly cleaved chlorite was investigated, which showed brucite-like and talc like surface areas. In constant amplitude FM mode, a slight variation in energy dissipation was observed between both surfaces. Amplitude and frequency vs. distance curves revealed that the tip was in repulsive contact with the specimen during imaging. PMID- 19131170 TI - Treatment of advanced pancreatic cancer: from gemcitabine single agent to combinations and targeted therapy. AB - The prognosis of advanced pancreatic adenocarcinoma is still poor nowadays. Gemcitabine in monotherapy (30-min infusion) has been the standard of treatment during the last decade, and many clinical trials have failed to demonstrate an improvement in overall survival (OS) with the addition of different drugs to gemcitabine, including cetuximab and bevacizumab. Nevertheless, some modest but interesting advances have been provided by combinations such as gemcitabine erlotinib, gemcitabine-capecitabine and gemcitabine plus a platinum salt. In spite of this, survival results remain disappointing. Further research focused on new combinations, incorporating the new targeted therapies and identifying potential predictive factors of response are required to be able to offer effective tailored therapies to these patients. PMID- 19131171 TI - Is exercise an acceptable and practical therapy for people with or cured of cancer? A systematic review. AB - INTRODUCTION: Therapeutic exercise, although potentially beneficial, does not appear acceptable to many cancer patients. A greater understanding of the reasons for this is required. We have systematically reviewed the use of exercise in this group, identifying rates of uptake, adherence and completion along with factors influencing acceptability. METHODS: Searches were completed using relevant key words. Data on study design, patient group, exercise intervention, patient flow data and reasons for declining or withdrawing from a programme were independently extracted by two researchers. Rates of study uptake, completion and adherence were compared according to dichotomised patient or programme characteristics using Mann Whitney U test (p=0.05). Reasons provided when declining or withdrawing from a study were categorised. RESULTS: Sixty five studies were included. The majority contained groups made up entirely or predominantly of patients with breast cancer offered an aerobic or resistance exercise programme. The median [IQR] rates of uptake, adherence and completion were 63 [33-80]%, 84 [72-93]% and 87 [80-96]%, respectively. No characteristic influenced the proportion of patients taking up or completing a programme. The main reasons for refusal were disinterest or the impracticality of undertaking the programme and for withdrawal, medical complication or deterioration. CONCLUSIONS: The invitation to undertake a programme of therapeutic exercise is accepted by about two-thirds of patients. Rates of adherence and completion are relatively high, but overall, only about half of patients offered a programme complete one. There is a need to modify exercise programmes if they are to be acceptable for the majority of patients. PMID- 19131172 TI - Putting a finger on potential predictors of oral contraceptive side effects: 2D:4D and middle-phalangeal hair. AB - Many women experience emotional or physical side effects when taking oral contraceptives (OCs). Despite the potential impact on women's health and well being, there are no valid methods to screen women for their risk of OC side effects. The present paper presents the results of two studies where anthropometric indicators of androgen exposure, 2D:4D and middle-phalangeal hair, were examined for their potential as predictors of OC side effects. In study 1, 2D:4D was associated with women's reports of a history of: (a) negative mood side effects; (b) discontinuation due to negative mood side effects; (c) specific mood side effects (i.e., crying, sadness, and altered trust in one's partners) and (d) specific physical side effects (i.e., headaches, fatigue, and decreased sex drive). In study 2, 2D:4D and/or middle-phalangeal hair was/were associated with a reported history of: (a) discontinuation due to negative mood side effects; (b) specific mood-related side effects (i.e., negative mood, disrupted sleep, increased aggression, and altered trust in one's partner) and (c) specific physical side effects (i.e., headaches, decreased menstrual cramps, and increased sex drive/arousal). The general pattern was that adverse OC side effects were experienced by women with lower 2D:4D and fewer middle-phalangeal hairs. Almost all relationships remained significant when response bias was controlled. These results suggest a possible role for prenatal testosterone exposure and both androgen action and sensitivity in women's experience of OC side effects. Furthermore, these two digit measures may be useful predictors of hormonal contraceptive side effects in women. PMID- 19131174 TI - Intralesional injection of Pingyangmycin may be an effective treatment for epulis. AB - Gingival epulis represents a family of benign tumors and tumorlike masses of the gingiva. A spectrum of inflammatory and other reactive changes can be observed pathologically as granulation, fibrous or vascular tissue. It may occur at any age, but most commonly found in patients at their twenties and sixties. Women are more frequently affected than men. Surgical excision or removal is the treatment of choice, and wider resection including extraction of the involved teeth and adjacent tissues is often required in order to prevent recurrence. However, wider resection may result in cosmetic and functional problems. Epulis bears some similarities with hemangioma both clinically and histopathologically. Therefore, we hypothesize that intralesional injection of Pingyangmycin may be a safe and effective treatment for epulis. The efficacy of this treatment modality is worthy of further investigation. PMID- 19131175 TI - Chronic organophosphate (OP)-induced neuropsychiatric disorder is a withdrawal syndrome. AB - Chronic organophosphate-induced neuropsychiatric disorder is a less well characterized syndrome, which is usually delay-occurred, persists long and is similar to the symptom of cholinergic deficit, its mechanism is unclear. The characteristics of chronic organophosphate-induced neuropsychiatric disorder are somewhat opposite to the direct action of OP pesticide, since withdrawal effect is usually opposite to the original effect of a drug, hypothesis that chronic organophosphate-induced neuropsychiatric disorder is a kind of withdrawal syndrome is suggested. PMID- 19131173 TI - Olanzapine shifts the temporal relationship between the daily acrophase of serum prolactin and cortisol concentrations rhythm in healthy men. AB - Treatment with the atypical antipsychotic drug olanzapine is frequently associated with development of obesity and insulin resistance. Treatment-induced weight gain has been suggested to be the main contributing factor of diminished insulin sensitivity. This study evaluated the effects of short-term treatment with olanzapine on 12h plasma prolactin and cortisol concentrations in healthy men. The effects of two distinct olanzapine formulations were investigated; the oral standard tablets (OST) and the orally disintegrating tablets (ODT). Recent reports indicate that treatment with the ODT formulation may be less harmful in terms of weight gain than the OST. 12 healthy men (age: 25.1+/-5.5 y) received olanzapine OST (10mg QD, 8 days), olanzapine ODT (10mg QD, 8 days) or no intervention in a randomized cross-over design. On day 8, blood samples were taken every 10min between 0000 and 1200h for determination of cortisol and prolactin concentrations. Treatment with olanzapine OST and ODT similarly increased the 12h mean PRL concentrations and the secreted PRL mass. Both drugs similarly shifted the maximal PRL concentration approximately 3-4h backwards in time. Cortisol secretions rates were lower, but the timing of the cortisol acrophase did not change. Both drugs significantly elevated HOMA index for insulin resistance. In conclusion olanzapine OST and ODT equally elevated the prolactin concentration and significantly shifted its acrophase, thus dissociating PRL and cortisol, while both formulations induced similar insulin resistance as evidenced by the elevated HOMA-IR. Notably, these alterations occurred without a measurable effect on body adiposity. PMID- 19131176 TI - Neurogenic hypothesis of cardiac ischemic pain. AB - Coronary heart disease is the major cause of death worldwide and is affecting millions of people in both developed and developing countries. Patients with myocardial ischemia typically experience chest pain (angina pectoris). Traditional viewpoint of ischemic cardiac pain might be related to "mechanical hypothesis" in early time and "chemical hypothesis" in modern time. However, perception of cardiac ischemic pain is still not well understood. The previous studies suggested that neurogenic mechanisms including neurogenic inflammation and neurogenic activity might participate in the pathophysiological processes following myocardial ischemia. Therefore, we raise "neurogenic hypothesis", that is, neurogenic mechanisms might play a pivotal role in myocardial ischemic injury. Analgesia intervention, rivalry of neurogenic inflammatory reactions and electrostimulatory therapy, etc. could not only relieve the pain symptoms, but also block nociception of body and neurogenic reaction induced by ischemia. Thereby ischemic myocardial injury would be extenuated and myocardial protection be produced. Attempts to confirm this hypothesis may lead to new theory of pathophysiologic mechanisms and provide potential intervention strategy for cardiac ischemic pain. PMID- 19131177 TI - Cognitive appraisals and physical health in people with posttraumatic stress disorder (PTSD). AB - Previous literature suggests that posttraumatic stress disorder (PSTD) is associated with poor physical health. To date a number of existing hypotheses have been developed to explain this association focusing predominantly on the interplay between cognitive and emotional appraisals. We have attempted to synthesise existing literature on the role of cognitive and emotional appraisals in explaining ill health (i.e. medically explained symptoms), following PTSD. On the basis of this review, we are introducing a conceptualisation aiming to explain poor physical health following PTSD. This new conceptualisation proposes that PTSD symptoms will lead to the production of two types of cognitive appraisals: Appraisals about the body that occurred during traumatisation, and cognitive appraisals about PTSD symptoms themselves. Both these cognitive appraisals coupled with negative emotional responses will lead to physiological arousal and negative health behaviours and subsequently to poor health. Although this conceptualisation has certain clinical implications for the management of physical health following PTSD, further research is required to test such by investigating the association between its elements. PMID- 19131178 TI - Autologous external jugular vein: a potential vascular graft for carotid artery reconstruction. AB - Carotid artery reconstruction is usually performed via interposition of a bridge between the vessel stumps by using autologous or artificial vascular grafts. The greater saphenous vein (GSV) and superficial femoral artery are most commonly used for this purpose, but many disadvantages exist when using these grafts. Recent studies has successfully reconstructed the carotid artery with EJV in sheep and found more smooth muscle cells in the tunica media of human EJV arranged circumferentially. In this paper, we hypothesize that autologous external jugular vein is more suitable for reconstruction of human carotid artery according to previous research evidences. Further clinical investigation is needed to determine the feasibility and reliability of using EJV for carotid reconstruction. PMID- 19131179 TI - Hypofractionated intensity-modulated radiotherapy for carcinoma of the prostate: analysis of toxicity. AB - PURPOSE: Dose escalation for prostate cancer improves biological control but with a significant increase in late toxicity. Recent estimates of low alpha/beta ratio for prostate cancer suggest that hypofractionation may result in biological advantage. Intensity-modulated radiotherapy (IMRT) should enable dose escalation to the prostate while reducing toxicity to local organs. We report late toxicity data of a hypofractionated IMRT regime. METHODS AND MATERIALS: Eligible men had T2-3N0M0 adenocarcinoma prostate, and either Gleason score >or= 7 or prostate specific antigen 20-50 ng/L. Patients received 57-60 Gy to prostate in 19-20 fractions using five-field IMRT. All received hormonal therapy for 3 months before radiotherapy to a maximum of 6 months. Toxicity was assessed 2 years postradiotherapy using the RTOG criteria, LENT/SOMA, and UCLA prostate index assessment tools. RESULTS: Acute toxicity was favorable with no RTOG Grade 3 or 4 toxicity. At 2 years, there was 4% Grade 2 bowel and 4.25% Grade 2 bladder toxicity. There was no Grade 3 or 4 bowel toxicity; one patient developed Grade 3 bladder toxicity. UCLA data showed a slight improvement in urinary function at 2 years compared with pretreatment. LENT/SOMA assessments demonstrated general worsening of bowel function at 2 years. Patients receiving 60 Gy were more likely to develop problems with bowel function than those receiving 57 Gy. CONCLUSIONS: These data demonstrate that hypofractionated radiotherapy using IMRT for prostate cancer is well tolerated with minimal late toxicity at 2 years posttreatment. Ongoing studies are looking at the efficacy of hypofractionated regimes with respect to biological control. PMID- 19131180 TI - Generic planning target margin for rectal cancer treatment setup variation. AB - PURPOSE: To calculate the generic planning target margin (GPTM) for patients receiving radiation therapy (RT) for rectal cancer placed in a prone position with a customized cradle for small-bowel exclusion. METHODS AND MATERIALS: A total of 25 consecutive rectal cancer patients were treated for 25 or 28 fractions in a prone position using a cradle to maximize small bowel exclusion. Treatment planning computed tomography (CT) scans were used to create orthogonally digitally reconstructed radiographs (DRRs) for portal image registration, which were compared with daily portal images from an electronic portal-imaging device (EPID). Translation values needed to align the DRRs and EPIDs were recorded for the superior to inferior (SI), right to left (RL), and anterior to posterior (AP) directions, and used to calculate the GPTM using the four-parameter model. Age, weight, and body mass index were tested compared with the setup variation using a Pearson correlation and a t test for significance. Gender versus setup variation was compared with a t test. RESULTS: A total of 1,723 EPID images were reviewed. The GPTM was 10 mm superior, 8 mm inferior, 7 mm RL and 10 mm AP. Age and gender were unrelated to setup variation. Weight was significantly associated with systematic AP variation (p < 0.05). BMI was significantly associated with systematic SI (p < 0.05) and AP (p < 0.01) variation and random RL variation (p < 0.05). CONCLUSIONS: The GPTM for rectal cancer is asymmetric with a maximum of 10 mm in the superior, anterior and posterior dimensions. Body mass index may effect setup variation. Research using advanced treatment planning should include these margins in the planning target volume definition. PMID- 19131181 TI - Defining the risk of involvement for each neck nodal level in patients with early T-stage node-positive oropharyngeal carcinoma. AB - PURPOSE: To assess the risk of ipsilateral subclinical neck nodal involvement for early T-stage/node-positive oropharyngeal squamous cell carcinoma. METHODS AND MATERIALS: Patients undergoing multilevel upfront neck dissection (ND) at Johns Hopkins Hospital within the last 10 years for early clinical T-stage (cT1-2) node positive (cN+) oropharyngeal squamous cell carcinoma were identified. Pathologic involvement of Levels IB-V was determined. For each nodal level, the negative predictive value of imaging results was computed by using sensitivity/specificity data for computed tomography (CT). This was used to calculate 1 - negative predictive value, or the risk that a negative level on CT harbors subclinical disease. RESULTS: One hundred three patients met the criteria. Radical ND was performed in 14.6%; modified radical ND, in 70.9%; and selective ND, in 14.6%. Pathologic positivity rates were 9.5%, 91.3%, 40.8%, 18.0%, and 3.3% for Levels IB-V, respectively. Risks of subclinical disease despite negative CT imaging results were calculated as 3.1%, 76.3%, 17.5%, 6.3%, and 1.0% for Levels IB-V, respectively. CONCLUSIONS: Levels IB and V are at very low (<5%) risk of involvement, even with ipsilateral to pathologically proven neck disease; this can guide radiation planning. Levels II and III should be included in high-risk volumes regardless of imaging results, and Level IV should be included within the lowest risk volume. PMID- 19131182 TI - First author research productivity of United States radiation oncology residents: 2002-2007. AB - PURPOSE: Participation in investigative research is a required element of radiation oncology residency in the United States. Our purpose was to quantify the first author research productivity of recent U.S. radiation oncology residents during their residency training. METHODS AND MATERIALS: We performed a computer-based search of PubMed and a manual review of the proceedings of the annual meetings of the American Society for Therapeutic Radiology and Oncology to identify all publications and presented abstracts with a radiation oncology resident as the first author between 2002 and 2007. RESULTS: Of 1,098 residents trained at 81 programs, 50% published > or =1 article (range, 0-9), and 53% presented > or =1 abstract (range, 0-3) at an American Society for Therapeutic Radiology and Oncology annual meeting. The national average was 1.01 articles published and 1.09 abstracts presented per resident during 4 years of training. Of 678 articles published, 82% represented original research and 18% were review articles. Residents contributed 15% of all abstracts at American Society for Therapeutic Radiology and Oncology annual meetings, and the resident contribution to orally presented abstracts increased from 12% to 21% during the study period. Individuals training at programs with >6 residents produced roughly twice as many articles and abstracts. Holman Research Pathway residents produced double the national average of articles and abstracts. CONCLUSION: Although variability exists among individuals and among training programs, U.S. radiation oncology residents routinely participate in investigative research suitable for publication or presentation at a scientific meeting. These data provide national research benchmarks that can assist current and future radiation oncology residents and training programs in their self-assessment and research planning. PMID- 19131183 TI - Would larger radiation fields lead to a faster onset of pain relief in the palliation of bone metastases? AB - PURPOSE: Hemibody irradiation has been shown to relieve bony metastatic pain within 24-48 hours of treatment, whereas for local external beam radiation, onset of pain relief is 1-4 weeks after radiation. The primary objective of this study is to examine whether there is a relationship between the areas of radiation treatment and onset of pain relief. METHODS AND MATERIALS: From Jan 1999 to Jan 2002, a total of 653 patients with symptomatic bone metastases were treated with external beam radiation. Pain scores and analgesic consumption were recorded at baseline and Weeks 1, 2, 4, 8, and 12. The areas of radiation treatment for all patients were calculated, then correlated with the response and analyzed in various ways. We first compared pain score alone with mean radiation field size. Second, we combined pain score and analgesic consumption. Last, we implemented the International Consensus end points for pain score and analgesic intake. RESULTS: Assessment of 653 patients showed no significant correlation comparing pain scores alone with radiation field area, with the exception of Week 4 for partial responders. Again, no significant correlation was found when combining both analgesic intake and pain score against radiation field size. Even when implementing the International Consensus end point definitions for radiation response, the only significant correlation between radiation field size and response was observed in Week 2 for partial response. CONCLUSION: There was no statistical significance between mean areas of radiation treatment with the onset of pain relief. PMID- 19131184 TI - Caveolin-1 as a prognostic marker for local control after preoperative chemoradiation therapy in rectal cancer. AB - PURPOSE: Caveolin-1 is a protein marker for caveolae organelles and has an essential impact on cellular signal transduction pathways (e.g., receptor tyrosine kinases, adhesion molecules, and G-protein-coupled receptors). In the present study, we investigated the expression of caveolin-1 in patients with rectal adenocarcinoma and correlated its expression pattern with the risk for disease recurrences after preoperative chemoradiation therapy (CRT) and surgical resection. METHODS AND MATERIALS: Caveolin-1 mRNA and protein expression were evaluated by Affymetrix microarray analysis (n = 20) and immunohistochemistry (n = 44) on pretreatment biopsy samples of patients with locally advanced adenocarcinoma of the rectum, and were correlated with clinical and histopathologic characteristics as well as with 5-year rates of local failure and overall survival. RESULTS: A significantly decreased median caveolin-1 intracellular mRNA level was observed in tumor biopsy samples as compared with noncancerous mucosa. Individual mRNA levels and immunohistologic staining, however, revealed an overexpression in 7 of 20 patients (35%) and 17 of 44 patients (38.6%), respectively. Based on immunohistochemical evaluation, local control rates at 5 years for patients with tumors showing low caveolin-1 expression were significantly better than for patients with high caveolin-1 expressing carcinoma cells (p = 0.05; 92%, 95% confidence interval [95% CI] = 82 102% vs. 72%, 95% CI = 49-84%). A low caveolin-1 protein expression was also significantly related to an increased overall survival rate (p = 0.05; 45%, 95% CI 16-60% vs. 82%, 95% CI = 67-97%). CONCLUSION: Caveolin-1 may provide a novel prognostic marker for local control and survival after preoperative CRT and surgical resection in rectal cancer. PMID- 19131185 TI - Gleason Pattern 5 prostate cancer: further stratification of patients with high risk disease and implications for future randomized trials. AB - PURPOSE: To compare prostate-specific antigen (PSA) outcomes in a cohort of men with high-risk prostate cancer based on the presence or absence of any Gleason Grade 5 component (primary, secondary, or tertiary). METHODS AND MATERIALS: Our study cohort consisted of 312 men with T1c-T3N0M0 prostate cancer with Gleason Scores of 7 with tertiary Grade 5, 8, or 9-10 who underwent radical prostatectomy or external beam radiotherapy with or without androgen suppression therapy. Cox regression multivariable analysis was used to assess whether a difference existed in risk of PSA recurrence in men with Gleason Score of 9-10 compared with those with Gleason Score of 8 and 7 with tertiary Grade 5, adjusting for treatment, age, and known prostate cancer prognostic factors. RESULTS: After a median follow up of 5.7 years, men with a Gleason Score of 8 had a lower risk of PSA recurrence than those with a Gleason Score of 9-10 (hazard ratio, 0.74; 95% confidence interval, 0.52-1.05; p = 0.09). Conversely, men with a Gleason Score of 7 with tertiary Grade 5 had a similar risk of PSA recurrence compared with men with a Gleason Score of 9-10 (hazard ratio, 1.08; 95% confidence interval, 0.60-1.94; p = 0.81). Median times to PSA failure for men with Gleason Scores of 9-10, 7 with tertiary Grade 5, and 8 were 4.5, 5.0, and 5.4 years, respectively. CONCLUSIONS: Our results highlight the importance of further substratification of the high risk Gleason Score category of 8-10 into 8 vs. 9, 10, and 7 with tertiary Grade 5. PMID- 19131186 TI - Low-dose involved-field radiotherapy as alternative treatment of nodular lymphocyte predominance Hodgkin's lymphoma. AB - PURPOSE: Nodular lymphocyte predominance Hodgkin's lymphoma is a very rare disease, characterized by an indolent clinical course, with sometimes very late relapses occurring in a minority of all patients. Considerable discussion is ongoing on the treatment of primary and relapsed disease. PATIENTS AND METHODS: A group of 9 patients were irradiated to a dose of 4 Gy on involved areas only. RESULTS: After a median follow-up of 37 months (range, 6-66), the overall response rate was 89%. Six patients had complete remission (67%), two had partial remission (22%), and one had stable disease (11%). Of 8 patients, 5 developed local relapse 9-57 months after radiotherapy. No toxicity was noted. CONCLUSION: In nodular lymphocyte predominance Hodgkin's lymphoma, low-dose radiotherapy provided excellent response rates and lasting remissions without significant toxicity. PMID- 19131187 TI - Cetuximab in combination with capecitabine, irinotecan, and radiotherapy for patients with locally advanced rectal cancer: results of a Phase II MARGIT trial. AB - PURPOSE: To evaluate the safety and efficacy of preoperative radiotherapy (RT) in combination with cetuximab, capecitabine, and irinotecan in patients with locally advanced rectal cancer. METHODS AND MATERIALS: Patients with rectal cancer (clinical stage T3/4 or N+) were scheduled to receive cetuximab (400 mg/m(2) Day 1, 250 mg/m(2) Days 8, 15, 22, 29) in combination with weekly irinotecan 40 mg/m(2) and capecitabine 500 mg/m(2) twice daily (Days 1-38). RT was given to a dose of 50.4 Gy (45 + 5.4 Gy). Primary endpoint was toxicity, and antitumor activity as assessed by the pathologic complete remission (pCR) rate was a secondary endpoint. RESULTS: Fifty patients were enrolled; 88% showed T3 or T4 and 76% nodal-positive tumors with a median distance from the anal verge of 7.5 cm. The actual dose intensity was as follows (median/mean, %): cetuximab 100/92, irinotecan 100/91, capecitabine 100/89). Main adverse events Grades 2/3/4 were (National Cancer Institute common toxicity criteria version 3.0, %): leukocytopenia 6/2/2, nausea/vomiting 4/2/0, diarrhea 34/30/0, proctitis 26/2/0, elevation of liver transaminases 8/10/0, and acnelike skin rash 46/6/0. All patients underwent surgery, and no postoperative deaths occurred. Eighty-four percent underwent low-anterior resection, and 68% of the specimen exhibited moderate or good tumor regression, but only 4 patients had a pCR. CONCLUSION: Preoperative chemoradiation with cetuximab-CapIri-RT has manageable toxicity, with diarrhea being the most commonly observed adverse event. Nevertheless, the efficacy of this regimen with a pCR rate of only 8% was significantly lower than that observed in a previous Phase I trial. PMID- 19131188 TI - Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection in wild boars. AB - The Porcine Reproductive and Respiratory Syndrome (PRRS) is one of the economically most important swine diseases worldwide. The virus can be spread by viraemic and persistent infected pigs. Spread and infection are advanced in areas with high herd and population densities. As wild boars have been found seropositive for PRRSV in Germany, France, and the USA, exchange between wild and domestic pig populations may exist. However, comprehensive information on PRRSV infection in wild boars is presently not available. The aim of the current study was thus, to systematically study PRRSV infection in wild boars to provide information on spatiotemporal, host and viral effects. The study was based on 531 wild boars from 52 hunts in Germany (2004-2007). PRRSV infection was determined and strains (US/EU) were classified by PCR. A total of 15.9% of the wild boars were PRRSV-positive (US: 14.2%; EU: 6.2%), with remarkable effects of state (US: 5.1-46.2%; EU: 0-17.6%), season (0-36.5%) and tissue (lungs: 89%; tonsils: 11%). Prevalences did neither correlate with age or weight, nor with density of production units, domestic pigs or wild boars. Open reading frame (ORF) 1 sequences within EU- and US-strains did not differ among wild boar samples. Homologies between EU-samples/Lelystad-virus and US-samples/PRRSV-MLV virus were 99.3 and 97%, respectively. This is the first comprehensive evidence of PRRSV infection in wild boars. We conclude that there is only a weak relation between wild boar and domestic PRRSV infection. PMID- 19131189 TI - Isolation, identification, and complete genome sequence of an avian reticuloendotheliosis virus isolated from geese. AB - Naturally occurring lymphoreticular tumors in geese have been found from time to time in Taiwan, but their etiology has not been determined except through morphological descriptions. This study observed a reticuloendotheliosis virus (REV) infection occurring in a white Roman goose (Anser anser) farm in Yunlin, Taiwan in 2006. These geese showed growth-retarded and nodular lymphoma-like tumors in the liver, lung, kidney, and pancreas. Thirty blood samples were taken for REV detection and 21 (70%) of them contained REV genetic sequences using polymerase chain reaction (PCR). Virus isolation was attempted from 11 blood samples by inoculating the buffy coat onto DF1 cells. Nine (81%) REVs were isolated after three blind passages. The complete proviral sequence from one isolate was determined for phylogenetic analysis by direct sequencing using overlapping PCR products. The length of the provial genome is 8284 nucleotides. By comparing with other published REV complete sequences, the nucleotide percent identity ranged from 93.5% to 99.8% with most LTR varieties, ranging from 74.9% to 99.8%. The present isolated goose REV is most close to REV APC-566, a REV isolated from Attwater's Prairie chickens. PMID- 19131190 TI - Positive effects of Taxol pretreatment on morphology, distribution and ultrastructure of mitochondria and lipid droplets in vitrification of in vitro matured porcine oocytes. AB - This study was designed to determine the effects of Taxol pretreatment on the morphology, distribution and ultrastructure of mitochondria and lipid droplets in vitrified porcine oocytes matured in vitro. The result showed that: (1) the rate of normal mitochondria distribution in fresh group (92.85%) was significantly higher (P<0.05) than that in other three groups (toxicity, 72.48%; vitrification, 50.83%; Taxol+vitrification, 69.98%) and Taxol pretreatment significantly (P<0.05) increased the ratio of normal mitochondria distribution in vitrified oocytes; (2) lipid droplets in vitrified oocytes got cracked, resulting in a great number of smaller lipid droplets (diameter <5 microm). The number of lipid droplets (5-10 microm in diameter) in vitrified oocytes pretreated with Taxol was higher (P<0.05) than that in the oocytes without Taxol pretreatment (81.87+/ 13.63 vs. 64.27+/-13.72); (3) both toxicity and vitrification cause the difference in the ultrastructure of mitochondria and lipid droplets. Mitochondria were well maintained in the form of typical round and ellipse shape with smooth surface and clear outline and lipid droplets existed in the form of integrity in Taxol pretreatment group. In conclusion, Taxol pretreatment has positive effects on vitrified porcine oocytes matured in vitro in terms of morphology, distribution and ultrastructure of mitochondria and lipid droplets. PMID- 19131191 TI - Fixed-time artificial insemination in red deer (Cervus elaphus) in Argentina. AB - Synchronization of estrous and fixed-time artificial insemination (FTAI) was conducted during the reproductive season of 2008 (March-April) in a local red deer breeding farm in Argentina. Multiparous suckling hinds (n=38) were artificially inseminated following hormonal treatment (intravaginal sponge containing 100mg of medroxiprogesterone acetate). At the time of sponge removal (day 12) 250 IU of eCG and 500 microg of PGF2alpha were given to each hind. The FTAI was performed at 48-55 h after device removal with cryopreserved semen imported from New Zealand. Rectal-transcervical AI method (similar to that in cattle) was performed and semen was deposited within the uterine body (n=28) or the cervix (n=10). Pregnancy was diagnosed by means of ultrasonography 44 days after FTAI. The overall pregnancy rate was 36.8% (14/38). Percentage of does that became pregnant with intrauterine seminal deposition was 42.9% (12/28) whereas pregnancy rate in the hinds with intracervical AI was 20% (2/10; P=0.27). PMID- 19131192 TI - A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes. AB - The general method of porcine in vitro fertilization (IVF), involving the co culture of both gametes in a medium drop, is thought to be the main reason for the high incidence of polyspermy. The aim of this study was to reduce the polyspermic fertilization of porcine embryos during IVF by the modified swim-up method, based on general sperm swim-up technique. Within this design, a 70 microm pore sized cell strainer was used to separate the sperm pellet placed at the bottom of a tube from the mature oocytes placed within the upper region. The separation of gametes using this permeable barrier was to ensure that only motile sperm gained access to the oocytes. It was found that the rate of polyspermy was significantly lowered for the sperm preparations from three boar breeds in modified swim-up method when compared with that of the general microdrop method (p<0.05). However, the penetration rates were found to be similar in both methods for two boar breeds. The average occurrence of blastocysts with more total cell number was higher in the modified swim-up method, while no significant difference in blastocyst rates between the two IVF methods was observed. The frequency of normal diploid embryos was also significantly higher in the modified swim-up method and polyploidy was more frequently observed in microdrop method (p<0.05). Our results demonstrated that the modified swim-up IVF method could reduce polyspermic penetration, and consequently produce better quality and karyotypically normal embryos in porcine IVF. PMID- 19131193 TI - Use of genital inspection and female urine tests to detect oestrus in captive Asian elephants. AB - Captive Asian elephant (Elephas maximus) populations are decreasing due to low birth rates compared to wild elephants. Improving oestrous detection in female elephants is required to ensure successful mating in captive and semi-captive herds. Responsive behaviours of eight semi-captive bull elephants to the uro genital area (genital inspection test) or urinary pheromones (urine test) of 14 female elephants throughout the oestrous cycle were evaluated. Weekly blood samples were collected for 27 consecutive months (14 months for the genital inspection test and 13 months for the urine test) from female elephants to characterize the patterns of circulating progestagen. Responsive behaviours of bulls were compared between females in the follicular versus the luteal phase of the cycle. The sensitivity and specificity of the genital inspection test were 65% and 68%, while those of the urine test were 52% and 61%, respectively. The bulls showed significantly higher "genital inspection", "flehmen from genital area" and "trunk on back" behaviours during the genital inspection test, and "flehmen" behaviours during the urine test in oestrous than in non-oestrous females. In sum, this study showed that monitoring sexual behaviours of Asian elephant bulls towards females or their urine can be used to detect the oestrous period. Although the sensitivity and specificity of both tests were not as high as expected, still, these methods appear to be more efficient at detecting oestrous than traditional methods based on mahout estimations of female receptivity. The use of genital inspection and urine tests may lead to more successful matings and thus to creating self-sustaining populations of captive elephants in range countries. PMID- 19131194 TI - Hyperinsulinaemia, a key factor of the metabolic syndrome in postmenopausal women. AB - The metabolic syndrome (MetS) is a complex disorder combining obesity, hypertension, atherogenic dyslipidaemia and insulin resistance, a clustering of factors which markedly enhance the risk of developing cardiovascular disease (CVD) and type 2 diabetes. Main features of the MetS, which are found in many postmenopausal women, are increasing prevalence of insulin resistance and obesity (particularly visceral adiposity). Accordingly, a majority of postmenopausal women comply with criteria defining the MetS, and CVD is the first cause of morbidity/mortality in women, occurring even more frequently than in men. Moreover, obesity-related type 2 diabetes approaches pandemic proportions. Simultaneous occurrence of insulin resistance and obesity are most detrimental for metabolic health, and are also associated with increased oxidative stress, inflammatory and prothrombotic processes as well as with postmenopausal alterations in adipocytokine production. Hormone replacement therapy, provided the selected progestin does not antagonize estrogen action, may improve fat mass and distribution, dyslipidaemia and insulin sensitivity in postmenopausal women. PMID- 19131195 TI - Gene polymorphisms that may influence the biological effects of progestins. AB - Many of the biological actions of progestins depend on binding to intracellular receptors and through a long chain of events to subsequent stimulation of transcriptional activity and protein synthesis. This process requires at least a few hours in time and many different proteins called coregulators do play a role after binding to the receptor. Evidence for polymorphisms in the gene coding for the PR has been obtained and many studies have already attempted to show associations between particular polymorphisms and human diseases. However, at present no consistent and conclusive picture has emerged on clinically important associations. Studies on links between polymorphisms in genes coding for coregulators are just beginning. The second pathway, the so-called non-genomic actions, is related to rapid effects of progestins that occur within minutes. At this moment a number of different membrane bound receptors have been identified. No data are available yet on polymorphisms in genes coding for these proteins or to link any of these membrane receptors to specific human pathology. PMID- 19131196 TI - Simultaneous liquid chromatographic-electrospray ionization mass spectrometric quantification of 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) and its metabolites 3,4-dihydroxymethamphetamine, 4-hydroxy-3-methoxymethamphetamine and 3,4-methylenedioxyamphetamine in squirrel monkey and human plasma after acidic conjugate cleavage. AB - 3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a psychoactive drug with abuse liability and neurotoxic potential. Specimen preparation of a recently presented LC-MS assay with electrospray ionization for quantifying MDMA and its main metabolites in squirrel monkey plasma was modified to include acidic hydrolysis to obtain total 3,4-dihydroxymethamphetamine and 4-hydroxy-3-methoxy methamphetamine. Method re-validation for squirrel monkey plasma and full validation for human plasma showed selectivity for all analytes. Recoveries were > or = 71.0%. Changed specimen preparation or matrix did not affect accuracy or precision. No instability was observed after repeated freezing or in processed samples. Plasma MDMA and metabolites quantification, derived pharmacokinetic and toxicokinetic data and neurotoxicity research will benefit from this validated method. PMID- 19131197 TI - Physiological concentrations of anabolic steroids in human hair. AB - Doping with endogenous anabolic steroids is one of the most serious issues in sports today. The measurement of anabolic steroid levels in human hair is necessary in order to distinguish between pharmaceutical steroids and natural steroids. This is the first investigation into the physiological concentrations of anabolic steroids in human hair in Chinese subjects. A gas chromatography tandem mass spectrometry (GC/MS/MS) method was developed for the simultaneous identification and quantitation of five endogenous anabolic steroids (testosterone, epitestosterone, androsterone, etiocholanolone and dehydroepiandrosterone) in hair. After basic hydrolysis, hair samples were extracted with diethyl ether, derivatized and then detected using GC/MS/MS in the multiple-reaction monitoring mode (MRM). The one precursor/two product ion transitions for each anabolic steroid were monitored. The limits of detection for the five endogenous anabolic steroids were in the 0.1-0.2 pg/mg range. All analytes showed good linearity and the extraction recoveries were 74.6-104.5%. Within-day and between-day precisions were less than 20%. This method was applied to the analysis of testosterone, epitestosterone, androsterone, etiocholanolone, and dehydroepiandrosterone in human hair. Full-length hair samples were taken at the skin surface from the vertex of 39 males, 30 females and 11 children from China. None of the subjects were professional athletes. Testosterone and dehydroepiandrosterone were detected in all the hair segments. The physiological concentrations of testosterone were in the range 0.8-24.2 pg/mg, 0.1-16.8 pg/mg and 0.2-11.5 pg/mg in males, females and children, respectively, however, the mean values of dehydroepiandrosterone were much higher than the concentrations of testosterone. These data are suitable reference values and are the basis for the interpretation of results from investigations into the abuse of endogenous anabolic steroids. PMID- 19131198 TI - Death of a female cocaine user due to the serotonin syndrome following moclobemide-venlafaxine overdose. AB - To our knowledge, the majority of evidence supporting the relationship between the serotonin syndrome and medications that effect 5HT is based on case reports. The justification for taking up this subject has been a fatal outcome of a 21 year-old female following an administration of toxic doses of moclobemide (MAOI) and venlafaxine (SNRI). As a result of complex toxicological investigations including antemortem and postmortem material, antemortem clinical observations and postmortem examinations, the cause of death was identified as overdose with antidepressants--moclobemide and venlafaxine--in the mechanism of the clinically fully developed severe toxic serotonin syndrome. The analysis of a hair strand collected from the victim documented the use of the above-mentioned drugs simultaneously with cocaine in the period of at least 20 months preceding death. The fact is a matter of considerable interest in view of the employed pharmacotherapy, giving rise to suspicion that the woman had not developed the serotonin syndrome during the almost 2-year antemortem period until she took toxic doses of both medications. PMID- 19131199 TI - The long-term effects of febrile seizures on the hippocampal neuronal plasticity clinical and experimental evidence. AB - Febrile seizures are the most common seizure disorder in childhood, but their long-term effects on the developing brains especially neuronal injury and neurocognitive function remain unresolved. Recent epidemiological studies reassure that most febrile seizures do not adversely affect global intelligence and hippocampal function, such as memory. However, there are concerns regarding those children who experience febrile seizures during the first postnatal year, having prior developmental delay and pre- or peri-natal events. Magnetic resonance imaging (MRI) studies confirmed that prolonged and focal FS can occasionally produce acute hippocampal injury that evolves into atrophy. Animal studies have revealed that the exposure of hippocampal neurons to experimental febrile seizures early in life, particularly prolonged or frequently repetitive FS, or together with brain malformation, may lead to sustained dysfunction of these cells, in spite of the absence of neuronal damage. Genetic studies suggest that the relationship between febrile seizures and subsequent epilepsy and neurocognitive dysfunction is sometimes genetic, but there are complex interactions with genetic or environmental modifiers. Therefore, there is a small group of children in whom febrile seizures-induced hippocampal injury might occur. Identification of the target population for subsequent mesial temporal sclerosis is important for prevention and early intervention. PMID- 19131200 TI - Crohn's disease: clinical-surgical questions and imaging answers. AB - Crohn's disease is an inflammatory bowel disease (IBD) characterized by a relapsing clinical pattern that typically affects people during their adult and economically productive lives. Affected patients require clinical follow-up because of the periodic flare-up of the disease and of the risk of long-term complications. Extensive diagnostic procedures, medical and surgical treatments are often needed over a lifetime. The challenge posed by the management of IBD is better faced by a multidisciplinary team that includes health care providers with complementary diagnostic or therapeutic skills. The team is expected to provide the best practice to manage IBD by defining a realistic "diagnostic and therapeutic pathway" for the patients to follow based on the locally available professional, structural and technological resources. For such a "pathway" the correct questions and answers are essential. Sometimes it is not easy to make sense out of these questions. To ask a right question is not simple. Of course, different surgeons and gastroenterologists ask different questions. If radiologists want to choose the right imaging method, they must know these questions. There exist a simple equation: good question=correct imaging method=right answer. PMID- 19131201 TI - Assessment of dynamic contrast enhancement of the small bowel in active Crohn's disease using 3D MR enterography. AB - PURPOSE: To retrospectively compare the dynamic contrast enhancement of the small bowel segments with and without active Crohn's disease at 3D MR enterography (MRE). MATERIALS AND METHODS: Thirteen patients (five men, eight women; mean age 41.2 years; range 29-56) were imaged on a 1.5-T MR scanner (Sonata, Siemens Medical) with standard MR sequences after having ingested 1000 ml of a 3% mannitol solution. Subsequently, high resolution 3D gradient-echo (volumetric interpolated breath-hold examination=VIBE) data sets were obtained pre-contrast and 20-40s, 60-80s, and 120-140 s after i.v. Gd-DOTA administration (0.2 mmol/kg). Signal enhancement was measured on single slices both in normal and histologically confirmed (12/13) inflamed small bowel wall segments as well as in the aorta, the psoas muscle, and the background to calculate signal-to-noise (SNR) and contrast-to-noise ratios (CNR). RESULTS: Small bowel wall enhancement was significantly higher (p<0.05) in inflamed compared to normal segments at 20 40s (SNR inflamed: 58.7+/-33.8 vs normal: 36.0+/-19.8; p=0.048; CNR inflamed: 34.8+/-23.4 vs normal: 16.3+/-11.2; p=0.017) and at 60-80s (SNR: 60.3+/-25.1 vs 41.9+/-20.0; p=0.049; CNR: 34.9+/-15.1 vs 19.3+/-13.2; p=0.01) after i.v. contrast administration, respectively. Even at 120-140 s CNR was still increased in inflamed segments (33.7+/-16.0 vs 18.1+/-13.2; p=0.04), while differences in SNR did not attain statistical significance (63.0+/-26.2 vs 45.3+/-23.3; p=0.15). CONCLUSION: In active Crohn's disease, histologically confirmed inflamed small bowel wall segments demonstrate a significantly increased early uptake of gadolinium on 3D VIBE sequences compared to normal small bowel segments. PMID- 19131202 TI - Isolation and structural elucidation of two impurities from a diacerein bulk drug. AB - Two impurities were found in the crude sample of diacerein. The level of these impurities 1.14% and 1.24% were detected by isocratic reverse-phase high performance liquid chromatography (HPLC). The molecular weights of the impurities were determined by liquid chromatography-mass spectroscopy (LC-MS) analysis. These impurities were isolated from crude sample of diacerein by reverse-phase preparative liquid chromatography. These impurities were characterized as 5 acetoxy-4-hydroxy-9,10-dioxo-9,10-dihydroanthracene-2-carboxylic acid (Impurity 1) and 4-acetoxy-5-hydroxy-9,10-dioxo-9,10-dihydroanthracene-2-carboxylic acid (Impurity-2) respectively. Structural elucidation of both the impurities were carried out by (1)H NMR, (13)C NMR, DEPT, 1D NOESY, MS and IR spectroscopy. PMID- 19131203 TI - Quantitative determination of famotidine polymorphs: X-ray powder diffractometric and Raman spectrometric study. AB - X-ray powder diffractometric and Raman spectrometric methods were developed for quantitative measurement of the polymorphic forms of famotidine in their mixtures. This study aims to deduce some useful conclusions regarding quantitative polymorph analysis, which could also be utilized in industrial practice. Both form A and form B of famotidine possess specific X-ray diffraction reflections as well as characteristic Raman vibrational bands, which permits simple determination of the phases in their mixtures. Keeping in mind that multivariate data processing by chemometric approach is thought of nowadays as superior over univariate one, the results of the two evaluation methods were compared by precision, accuracy as well as robustness. It was found that both approaches provide similar results provided analytically useful data regions are properly selected. Overcoming the common problems of quantitative X-ray powder diffractometry and solid state Raman spectrometry both permit accurate quantification of famotidine polymorphs; the latter, however, seems to be more favourable in regular laboratory practice. PMID- 19131204 TI - LC method for the therapeutic drug monitoring of lamotrigine: evaluation of the assay performance and validation of its application in the routine area. AB - An accurate and precise high-performance liquid chromatographic method using diode array detection for the determination of lamotrigine in human plasma has been developed and validated for use in pharmacokinetic studies. A validation strategy based on the accuracy profiles was used to select the most appropriate regression model and to determine the limits of quantitation as well as the concentration range. On the other hand, the present paper also shows this validation approach as a suitable tool to guaranty the quality of the results obtained by the use of the analytical validated methodology for plasma lamotrigine determination in a routine setting and to ensure the risk of obtaining the future measurements outside the previously fixed acceptance limits. PMID- 19131205 TI - Patterns of recurrence after curative resection of pancreatic ductal adenocarcinoma. AB - AIMS: Despite curative surgery for pancreatic ductal adenocarcinoma (PDAC), most patients develop cancer recurrence and die from metastatic disease. Understanding of the patterns of failure after surgery can lead to new insights for novel therapeutic modalities. The aim of the present study is to describe the patterns of recurrence after curative resection of PDAC. METHODS: A retrospective analysis was performed of 145 consecutive resections for PDAC between 1998 and 2005 (M/F 75/70; median (range) age 67 years (32-85 y)). The location of the first and consecutive recurrences, and the time interval to cancer recurrence after surgical resection was studied. The magnitude of tumour-free margin was less than a millimetre in 48 patients, whereas a positive surgical margin was observed in 27 patients. The median duration of follow-up was 18.5 (range 0.3-116.8) months. RESULTS: Cancer recurrence was observed in 110 patients. The first location of recurrence was locoregional in 19, extra-pancreatic in 66, and combined locoregional and extra-pancreatic in 25 patients. Extra-pancreatic recurrence developed in the liver in 57, peritoneal in 35, pulmonary in 15, and retroperitoneal in 5 patients. The median (95% CI) overall (OS) and disease-free (DFS) survival was 18.7 (15.7-23.5) and 9.8 (7.5-12.4) months, respectively. The type of cancer recurrence did not significantly influence OS, while the resection margin status had a prognostic effect. CONCLUSION: The vast majority of patients who undergo potentially curative surgery for PDAC develop cancer recurrence located in the abdominal cavity. Surgical resection margins with tumour involvement and tumour-free margins of less then 1mm are negative prognostic factors. Further research on better local surgical control, peri-operative locoregional treatment, and more effective adjuvant systemic therapy is necessary to improve long-term survival of patients with curable PDAC. PMID- 19131206 TI - [Long texts for the formalized recommendation of experts on management of postoperative pain]. PMID- 19131207 TI - Intrapericardial paraganglioma with intratumoral coronary arterial aneurysm and an arteriovenous fistula. AB - The authors present the case of a 55-year-old woman with a nonfunctioning intrapericardial paraganglioma. The tumor was incidentally found during transthoracic echocardiography that was performed because an enlarged cardiac silhouette had been noted on a chest x-ray. Transesophageal echocardiography further elucidated that the left circumflex coronary artery branched and traversed into the intrapericardial mass. Coronary angiography and cardiac multislice computed tomography both confirmed the presence of a vascular intrapericardial mass being fed by the obtuse marginal artery. Moreover, an arterial aneurysmal formation and a coronary arteriovenous fistula were noted within the tumor. After "en bloc" tumor resection, histopathologic and electron microscopic examinations established the diagnosis of a nonfunctioning intrapericardial paraganglioma. PMID- 19131208 TI - Intrinsic myoarchitectural differences between the left and right ventricles of fetal human hearts: an ultrasonic backscatter feasibility study. AB - OBJECTIVE: Embryologically, cardiac chambers differ in their morphologic and contractile properties from the beginning. We hypothesized that a noninvasive ultrasonic backscatter investigation might illustrate the fundamental differences in myocardial morphologic properties of the 2 ventricles during heart development. The goals of this investigation were to 1) explore the feasibility of measuring the magnitude of cyclic variation of ultrasonic backscatter from the left and right ventricular free walls of fetal hearts; 2) compare measurements of the magnitude of cyclic variation from the left and right sides of the heart; and 3) determine if the observed results are consistent with predictions relating the overall backscatter level and the magnitude of cyclic variation. METHODS: Cyclic variation data from the left and right ventricular free walls were generated from analyses of the backscatter from echocardiographic images of 16 structurally normal fetal hearts at mid-gestation. RESULTS: The magnitude of cyclic variation was found to be greater for the left ventricular free wall than for the right ventricular free wall (4.5 +/- 1.1 dB vs 2.3 +/- 0.9 dB, respectively; mean +/- standard deviation; P < .0001, paired t test). CONCLUSION: Measurements of the cyclic variation of backscatter can be obtained from both the left and right sides of fetal hearts demonstrating a significant difference between the measured magnitude of cyclic variation in the left and right ventricular myocardium. This observation is consistent with predictions relating the overall backscatter level and the magnitude of cyclic variation. The results of this study suggest cyclic variation measurements may offer a useful approach for characterizing intrinsic differences in myocardial properties of the 2 ventricles in assessing fetal heart development. PMID- 19131209 TI - A unifying framework for understanding heart failure? Response to "Left Ventricular Torsion By Two-Dimensional Speckle Tracking Echocardiography in Patients With Diastolic Dysfunction and Normal Ejection Fraction" by Park SJ et al. PMID- 19131210 TI - Evaluation of hepatic perfusion and function with modified hepatobiliary scintigraphy. AB - The quantitative relationship between changes in portal vein pressure and the perfusion index was studied, and hepatocellular function was evaluated. A modified protocol of hepatobiliary dynamic scintigraphy with Tc-99m-labeled ethylene hepatobiliary iminodiacetic acid (EHIDA) was performed in 37 patients with hepatic cirrhosis and 12 healthy controls. In 18 patients, the portal vein pressure (PVP) was measured intra-operatively during the portal vein bypass procedure. The portal vein perfusion index (PVI) was obtained by a two compartment model of hepatic perfusion. A three-compartment model was applied in the hepatocellular extracted and excreted time-radioactivity curves, and the hepatic functional index was calculated including the hepatic uptake index (UI), the mean residual index (MRI), the uptake speed index (UsI), the descent speed index (DsI), the peak uptake (PU), and the peak time (PT) of hepatic uptake. These function indices in the cirrhotic patients were compared to the indices in the healthy controls with a two-sided t-test; specifically, the PU (18.94+/-6.80 vs. 29.67+/-18.98, P<0.01), UI (0.84+/-5.99 vs. 18.41+/-13.87, P<0.05), and UsI (0.58+/-0.39 vs. 1.23+/-0.81, P<0.01) were slower in patients with hepatic cirrhosis compared with the healthy controls. The PT (16.83+/-4.89 vs. 10.95+/ 2.79, P<0.001) and MRI (5.93+/-2.96 vs. 2.74+/-0.97, P<0.001) were more prolonged and the DsI (0.07+/-0.09 vs. 0.18+/-0.14, P<0.01) was less compared to controls. The PVI were significantly higher in cirrhotic patients than in controls (46.17+/ 7.83 vs. 29.07+/-5.71, P<0.001), and the PVI correlated to the PVP (r=0.79, P<0.01). In conclusion, modified quantitative hepatobiliary dynamic scintigraphy can evaluate the changes of portal vein blood flow and monitor hepatocellular function, in which the portal vein blood flow can estimate portal vein pressure. PMID- 19131211 TI - IP-10 protects while MIP-2 promotes experimental anesthetic hapten - induced hepatitis. AB - MIP-2 and IFN-gamma inducible protein-10 (IP-10) and their respective receptors, CXCR2 and CXCR3, modulate tissue inflammation by recruiting neutrophils or T cells from the spleen or bone marrow. Yet, how these chemokines modulate diseases such as immune-mediated drug-induced liver injury (DILI) is essentially unknown. To investigate how chemokines modulate experimental DILI in our model we used susceptible BALB/c (WT) and IL-4-/- (KO) mice that develop significantly reduced hepatitis and splenic T cell priming to anesthetic haptens and self proteins following TFA-S100 immunizations. We detected CXCR2+ splenic granulocytes in all mice two weeks following immunizations; by three weeks, MIP-2 levels (p<0.001) and GR1+ cells were elevated in WT livers, suggesting MIP-2-recruited granulocytes. Elevated splenic CXCR3+CD4+T cells were identified after two weeks in KO mice indicating elevated IP-10 levels which were confirmed during T cell priming. This result suggested that IP-10 reduced T cell priming to critical DILI antigens. Increased T cell proliferation following co-culture of TFA-S100-primed WT splenocytes with anti-IP-10 (p<0.05) confirmed that IP-10 reduced T cell priming to CYP2E1 and TFA. We propose that MIP-2 promotes and IP-10 protects against the development of hepatitis and T cell priming in this murine model. PMID- 19131212 TI - Delayed-onset muscle injury and its modification by wheat gluten hydrolysate. AB - OBJECTIVE: We investigated the pattern of delayed-onset muscle injury in well trained athletes after a competitive half-marathon and the effects of post-race intake of wheat gluten hydrolysate (WGH). METHODS: Thirty well-trained college runners with a best time of 14-15 min over 5000 m raced in a half-marathon. Thereafter, they were divided into three groups based on finish times and given 0 (control), 10, or 20 g of WGH. Blood biochemical parameters were monitored at -1 d, +1 h, +1 d, and +2 d after the race. Data selected according to finish times and biochemical parameters were then analyzed. RESULTS: Plasma creatine kinase activity peaked at 1 d after the race in the control group and correlated with post-race white blood cell counts. The post-race elevation of creatine kinase activity was dose-dependently suppressed by WGH. CONCLUSION: Delayed-onset muscle injury peaked in well-trained distance runners at 1 d after a half-marathon and was dose-dependently suppressed by a post-race intake of WGH. PMID- 19131213 TI - Relation of visfatin to cardiovascular risk factors and adipocytokines in patients with impaired fasting glucose. AB - OBJECTIVE: The discovery of visfatin has great potential to significantly enhance our understanding of impaired fasting glucose and diabetes mellitus. The aim of the present study was to explore the relation of visfatin concentrations to cardiovascular risk factors and serum adipocytokine concentrations in patients with impaired fasting glucose. METHODS: A sample of 55 patients with impaired fasting glucose was analyzed in a prospective way. All patients with a 2-wk weight-stabilization period before recruitment were enrolled. Weight, blood pressure, basal glucose, lipoprotein(a), C-reactive protein, insulin, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triacylglycerols, blood, and adipocytokines (visfatin, leptin, adiponectin, resistin, tumor necrosis factor-alpha [TNF-alpha], and interleukin 6) levels were measured. Tetrapolar impedancometry, indirect calorimetry, and prospective serial assessment of nutritional intake with 3-d written food records were performed. RESULTS: Fourteen men (25.5%) and 41 women (74.5%), with a mean age of 57.3 +/- 11.7 y and mean body mass index of 35.8 +/- 3.6 kg/m(2), were included. Patients were divided in two groups by median visfatin value (18.2 ng/mL): group I had low values and group II had high values. Patients in group I had greater weight, body mass index, fat mass, fat-free mass, and adiponectin than patients in group II. Patients in group II had higher total cholesterol, low density lipoprotein cholesterol, resistin, and TNF-alpha levels than patients in group I. In the multivariate analysis with age- and sex-adjusted basal visfatin concentration as a dependent variable, only TNF-alpha remained an independent predictor in the model (F = 8.4, P < 0.05), with an inverse correlation. Visfatin concentration decreased 7.33 ng/mL (95% confidence interval 2.10-12.58) for each nanogram per milligram of TNF-alpha increase. CONCLUSION: Only TNF-alpha is related in an independent way to serum visfatin levels. PMID- 19131214 TI - Beneficial effects of gradual intense exercise in tissues of rats fed with a diet deficient in vitamins and minerals: a pilot study. AB - OBJECTIVE: This study evaluated the preliminary effects of intense physical training (swimming) on oxidative stress in rats with nutritional deficiencies. METHODS: Rats were fed with a standard diet or a diet deficient in vitamins and minerals for 4 months. The deficient diet contained one-fourth of the recommended vitamin and mineral levels for rats. From the second month, half of the animals were subjected to a swimming exercise in a plastic container with water maintained at 34 +/- 1 degrees C for 1 h/d, five times per week, for 11 wk. The rats were subjected to swimming exercise with loads attached to the dorsal region, which were progressively increased according to their body weight (1% to 7%). Sedentary rats were transported to the experimental room and handled as often in a similar way as the exercise group, except that they were not put in water. RESULTS: In the exercised group, blood lactate levels were significantly lower and the heart weight/body weight ratio was significantly higher than in the sedentary group (P < 0.05). Increased lipid peroxidation was observed in the liver, heart, and skeletal muscle of rats fed with the deficient diet, but it was completely reversed by exercise. Exercise also decreased lipid peroxidation levels in the heart and skeletal muscle of rats fed with the standard diet (P < 0.05). CONCLUSION: This pilot study leads to the continuity of the studies, because the partial results observed suggest that inadequate nutrition may enhance oxidative stress, and that intense chronic physical training may activate antioxidant defenses, possibly by hormesis. PMID- 19131215 TI - Circulating ghrelin/obestatin ratio in subjects with Helicobacter pylori infection. AB - OBJECTIVE: Ghrelin is a peptide hormone involved in human energy homeostasis. Obestatin is a recently discovered active peptide derived from preproghrelin. It seemed that obestatin was a physiologic opponent of ghrelin. Helicobacter pylori infection may be associated with appetite and nutrition. We compared the plasma ghrelin/obestatin ratio in H. pylori-positive and -negative groups. METHODS: People undergoing an annual health checkup were included. Helicobacter pylori status was based on serologic and carbon-13 urea breath findings. Fifty adults with H. pylori infection and 50 adults matched by age and body mass index without H. pylori infection were enrolled in this study. Plasma ghrelin and obestatin levels were measured by radioimmunoassay. RESULTS: Ghrelin concentrations and ghrelin/obestatin ratios were lower in the H. pylori-positive group than in the H. pylori-negative group. There was no significant difference in circulating obestatin between those with and without H. pylori infection. In all subjects, the ghrelin/obestatin ratio was negatively correlated with body mass index, the homeostasis model of assessment for insulin resistance, and serum levels of triacylglycerol. There was a positive correlation between circulating obestatin and ghrelin levels. CONCLUSION: Helicobacter pylori infection was associated with a reduction in the circulating ghrelin/obestatin ratio in Chinese adults. PMID- 19131216 TI - Correlation NMR spectroscopy involving quadrupolar nuclei. AB - We review the recent developments proposed for integer or half-integer quadrupolar nuclei, focussing on the methods to observe them under high resolution and to analyze their through-space and through-bond connectivities. PMID- 19131217 TI - Graft copolymerization onto cellulose-based filter paper and its further development as silver nanoparticles loaded antibacterial food-packaging material. AB - The present work describes ceric ammonium nitrate (CAN) initiated graft copolymerization of acrylamide onto cellulose-based filter paper followed by entrapment of silver nanoparticles. The copolymerization was carried out in aqueous solution, containing 2M acrylamide monomer and 16mM N,N'-methylene bisacrylamide (MB) crosslinker. The optimum initiation time and grafting reaction temperature were found to be 15min and 30 degrees C, respectively. The silver nanoparticles were loaded into grafted filter paper by equilibration in silver nitrate solution followed by citrate reduction. The formation of silver nanoparticles has been confirmed by TEM and SAED analysis. The novel nano silver loaded filter paper has been investigated for its antimicrobial properties against E.coli. This newly developed material shows strong antibacterial property and thus offers its candidature for possible use as antibacterial food-packaging material. PMID- 19131218 TI - An efficient biosurfactant-producing bacterium Pseudomonas aeruginosa MR01, isolated from oil excavation areas in south of Iran. AB - A bacterial strain was isolated and cultured from the oil excavation areas in tropical zone in southern Iran. It was affiliated with Pseudomonas. The biochemical characteristics and partial sequenced 16S rRNA gene of isolate, MR01, was identical to those of cultured representatives of the species Pseudomonas aeruginosa. This bacterium was able to produce a type of biosurfactant with excessive foam-forming properties. Compositional analysis revealed that the extracted biosurfactant was composed of high percentages lipid ( approximately 65%, w/w) and carbohydrate ( approximately 30%, w/w) in addition to a minor fraction of protein ( approximately 4%, w/w). The best production of 2.1g/l was obtained when the cells were grown on minimal salt medium containing 1.2% (w/v) glucose and 0.1% (w/v) ammonium sulfate supplemented with 0.1% (w/v) isoleucine at 37 degrees C and 180rpm after 2 days. The optimum biosurfactant production pH value was found to be 8.0. The MR01 could reduce surface tension to 28mN/m and emulsified hexadecane up to E24 approximately 70. The results obtained from time course study indicated that the surface tension reduction and emulsification potential was increased in the same way to cell growth. However, maximum biosurfactant production occurred and established in the stationary growth phase (after 84h). Fourier Transform Infrared spectrum of extracted biosurfactant indicates the presence of carboxyl, amine, hydroxyl and methoxyl functional groups. Thermogram of biosurfactant demonstrated three sharp endothermic peaks placing between 200 and 280 degrees C. The core holder flooding experiments demonstrated that the oil recovery efficiencies varied from 23.7% to 27.1% of residual oil. PMID- 19131219 TI - Distribution of Sorbitan Monooleate in poly(L-lactide-co-epsilon-caprolactone) nanofibers from emulsion electrospinning. AB - The aim of this study was to investigate the distribution of Sorbitan Monooleate (Span80) in poly(l-lactide-co-epsilon-caprolactone) (PLLACL) nanofibers from emulsion electrospinning. The hypothesis was that PLLACL/Span80 nanofibrous mats would have some Span80 on the surface of the composite nanofibers. To test the hypothesis, the electrospinning of emulsions made of PLLACL, chloroform, Span80, and distilled water to prepare PLLACL/Span80 nanofibers was systematically investigated. The morphology of PLLACL/Span80 nanofibers was investigated by atomic force microscopy. The surface hydrophilicity of the nanofibrous mats were examined by water contact angle test. The distribution of Span80 on the surface of nanofibrous mats was also confirmed by the performance of pig iliac endothelium cells on the nanofibrous mats. PMID- 19131220 TI - Sub- and post-micellar catalytic and inhibitory effects of cetyltrimethylammonium bromide in the permanganate oxidation of phenylalanine. AB - The kinetics of phenylalanine (phe) oxidation by permanganate has been investigated in absence and presence of cetyltrimethylammonium bromide (CTAB) using conventional spectrophotometric technique. The rate shows first- and fractional-order dependence on [MnO(4)(-)] and [phe] in presence of CTAB. At lower values of [CTAB] (< or =10.0x10(-4)moldm(-3)), the catalytic ability of CTAB aggregates are strong. In contrast, at higher values of [CTAB] (> or =10.0x10(-4)moldm(-3)), the inhibitory effect was observed in absence of H(2)SO(4). We find that anions (Br(-), Cl(-) and NO(3)(-)) in the form of sodium salts are strong inhibitors for the CTAB catalyzed oxidation. Kinetic and spectrophotometric evidences for the formation of an intermediate complex and an ion-pair complex between phe and MnO(4)(-), CTAB and MnO(4)(-), respectively, are presented. A mechanism consistent with kinetic results has been discussed. Complex formation constant (K(c)) and micellar binding constant (K(s)) were calculated at 30 degrees C and found to be K(c)=319mol(-1)dm(-3) and K(s)=1127mol(-1)dm(-3), respectively. PMID- 19131221 TI - Covalent conjugation of tetrameric bovine liver catalase to liposome membranes for stabilization of the enzyme tertiary and quaternary structures. AB - Tetrameric bovine liver catalase (BLC) is unstable because of its dissociation into subunits at low enzyme concentrations and the conformational change of the subunits at high temperatures. In this work, for stabilization of BLC, the enzyme was covalently conjugated with liposome membranes composed of 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine (POPC), cholesterol and 1,2-dioleoyl-sn glycero-3-phosphoethanolamine-N-glutaryl (NGPE). The NGPE, which was responsible for the BLC/membrane coupling, was altered from 0.05 to 0.2 in its liposomal mole fraction f(G). The catalase-conjugated liposome (CCL) with f(G) of 0.15 showed the maximum number of the conjugated BLC molecules of 28 per liposome. The reactivity of CCLs to H(2)O(2) was as high as that of free BLC at 25 degrees C in Tris-HCl buffer of pH 7.4. Among the CCLs, the catalyst with f(G) of 0.15 was the most stable at 55 degrees C in its enzyme activity in the buffer because the appropriate number of BLC/liposome covalent bonding prevented the dissociation induced enzyme deactivation. Furthermore, the CCL showed much higher stability at 55 degrees C than the free BLC/enzyme-free liposome mixture and free BLC at the low BLC concentration of 340ng/mL. This was because BLC in the CCL was located in the vicinity of the host membrane regardless of the catalyst concentration, which could induce the effective stabilization effect of the membrane on the enzyme tertiary structure as indicated by the intrinsic tryptophan fluorescence analysis. The results obtained demonstrate the high structural stability of BLC in the CCL system, which was derived from the covalent bonding and interaction between BLC and liposomes. PMID- 19131222 TI - Synthesis of Ag-Pt alloy nanoparticles in aqueous bovine serum albumin foam and their cytocompatibility against human gingival fibroblasts. AB - Foams of bovine serum albumin (BSA) have been utilized for synthesizing in situ protein capped Ag-Pt alloy nanoparticles. The in vitro cytotoxicity and the rate of proliferation of human gingival fibroblasts (HGFs) in presence of the above synthesized alloy nanoparticles is investigated. Expression profile of protein involved in detoxification, i.e. metallotheonein (MT) were assayed by ELISA and expression of mRNA transcripts by reverse transcription polymerase chain reaction (RT-PCR). Cytotoxicity results suggested that protein capped nano-alloys might be promising candidates for implants and prosthetic material. RT-PCR and ELISA confirmed the expression of MT, in cells treated with the alloy nanoparticles. Morphology variation studied by SEM also confirms that cells treated with alloy nanoparticles present an intact morphology. PMID- 19131223 TI - Preparation and characteristics of linoleic acid-grafted chitosan oligosaccharide micelles as a carrier for doxorubicin. AB - The linoleic acid (LA)-grafted chitosan oligosaccharide (CSO) (CSO-LA) was synthesized in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and the effects of molecular weight of CSO and the charged amount of LA on the physicochemical properties of CSO-LA were investigated, such as CMC, graft ratio, size, zeta potential. The results showed that these chitosan derivatives were able to self-assemble and form spherical shape polymeric micelles with the size range of 150.7-213.9nm and the zeta potential range of 57.9-79.9mV, depending on molecular weight of CSO and the charged amount of LA. Using doxorubicin (DOX) as a model drug, the DOX-loaded CSO-LA micelles were prepared by dialysis method. The drug encapsulation efficiencies (EE) of DOX-loaded CSO-LA micelles were as high as about 75%. The sizes of DOX-loaded CSO-LA micelles with 20% charged DOX (relating the mass of CSO-LA) were near 200nm, and the drug loading (DL) capacity could reach up to 15%. The in vitro release studies indicated that the drug release from the DOX-loaded CSO-LA micelles was reduced with increasing the graft ratio of CSO-LA, due to the enhanced hydrophobic interaction between hydrophobic drug and hydrophobic segments of CSO-LA. Moreover, the drug release rate from CSO-LA micelles was faster with the drug loading. These data suggested the possible utilization of the amphiphilic micellar chitosan derivatives as carriers for hydrophobic drugs for improving their delivery and release properties. PMID- 19131224 TI - Modulation of the physicochemical state of interior agents to prepare controlled release liposomes. AB - To prepare controlled release liposomes, freeze-drying of double emulsions (FDE) was employed to entrap a model drug, topotecan, which has been reported to tend to leak rapidly from vesicles. In addition, hydrogenated soy phosphatidylcholine (HSPC), N-(carbonyl-methoxypolyethyleneglycol2000)-1,2-distearoyl-sn-glycero-3 phosphoethanolamine (PEG-PE), and cholesterol (3:1:1, mass ratio) were used as the vesicle lipid. Different inner aqueous phases (W1) containing topotecan together with a variety of chemicals, such as citrate, sulfate, and divalent copper ions, were used to prepare W1/O/W2 double emulsions, which were then freeze-dried to obtain dry products. Upon rehydration, the dry products formed topotecan-entrapping unilamelar liposomes with an encapsulation efficiency of 80% and a mean diameter of less than 200nm. The in vitro release experiments demonstrated that the drug release of topotecan-entrapping FDE liposomes could be successfully controlled through altering the state of the incorporated drug by means of protonation, precipitation, or forming a transition metal-complex. Specifically, the formulation of 300mM CuSO(4) had a drug release half-life of 36h. This novel method is a promising way to prepare controlled release liposomes that are more suitable for therapeutic application. In addition, the liposome formation mechanism was discussed based on micrographs and the size of the double emulsions and vesicles, as well as the small angle X-ray scattering pattern and transmission electron microscopy images. PMID- 19131225 TI - Treadmill exercise reduces self-administration of morphine in male rats. AB - Exercise can activate the same pathways as morphine. The aim of the present study was to clarify the effect of short-term and mid-term exercises on the self administration of morphine in rats. Male Wistar rats were initially trained to receive small pellets of food by pressing the active lever in self-administration apparatus. Rats were divided into 4 groups: Saline, Morphine, Exercise 1 (11 days) and Exercise 2 (30 days). Their jugular vein was cannulated. The animals were placed in self-administration apparatus and allowed to self-administer morphine (0.5mg per infusion all test groups) or saline (Saline group) during consecutive days, for 2h/sessions. In the group 1 the rats were running before each session of self-administration and of group Exercise 2, 30 days before surgery as well as before each session. The pressing numbers of active and passive levers in each group and among different groups were compared. The number of active lever pressing of Morphine group was significantly higher than Saline group (p<0.001). In Exercise 1 and Exercise 2 groups, the number of active lever pressing was significantly lower than Morphine group (p<0.001). As exercise can activate many neurotransmitter systems involved in the addiction process and increase the release of endorphins, it is likely that could decrease the morphine self-administration in this experimental setup. PMID- 19131226 TI - The involvement of pro-inflammatory cytokines in nephrogenic systemic fibrosis - a mechanistic hypothesis based on preclinical results from a rat model treated with gadodiamide. AB - The evidence for the potential involvement of gadolinium-based contrast agents (GBCAs) in the pathomechanism of nephrogenic systemic fibrosis (NSF), a rare but serious disease occurring in patients with severe or end-stage renal failure, has grown due to recent epidemiological and preclinical research. Nevertheless there is still uncertainty with regard to the prevailing patho-physiological processes that may lead to NSF. To examine the potential mechanism of the fibrotic skin changes we applied a recently published rat model of NSF for investigations into serum markers for inflammation. For this purpose male Wistar rats were treated either once, three, or eight times with a daily intravenous injection of 2.5 mmol/kg gadodiamide, the drug substance of the magnetic resonance imaging (MRI) agent Omniscan. Clinical observations, hematology, clinical pathology, histopathology including electron microscopy and gadolinium (Gd) determination in serum, skin, femur and liver tissue, and a multiplexed analysis of 70 protein serum markers were performed. Gd was detectable in the skin, femur, and liver of the gadodiamide-treated rats 6h after the first administration. Macroscopic skin changes, appearing as reddening and early scab formation, were observed in one animal after the third daily administration and affected all animals after 8 daily administrations. Microscopy revealed dermal infiltrations after three administrations, progressing towards inflammatory lesions, ulcerations and crusts. Among the investigated serum marker panel 13 cytokines were significantly (p<0.01) elevated 6 h after the first injection, and eight stayed elevated over all time points: the monocyte chemotactic proteins MCP-1 and MCP-3, the macrophage inflammatory proteins MIP-1beta and MIP-2, the tumor necrosis factor TNF-alpha, the extracellular matrix regulator tissue inhibitor of metalloproteinase type 1 (TIMP-1), the vascular epithelial growth factor (VEGF) and osteopontin. The latter cytokine is of particular interest, since this matrix cellular glycoprotein is involved in the regulation of dystrophic calcification but also plays a role as a chemoattractant for dendritic cells, macrophages and T lymphocytes, which in turn activate inflammatory pathways. Reflecting the physiological role of osteopontin, we hypothesize that Gd release from the GBCA complex leads to the formation of insoluble Gd-deposits subsequently eliciting a physiological response similar to that seen during dystrophic calcification, i.e. an up-regulation of osteopontin and chemoattractant cytokines. Concomitant increase in vascular permeability caused by MIP-1, TNF-alpha and VEGF may lead to extravasation of chelated Gd or Gd-deposits. The inherent persistence of the Gd deposits may subsequently result in an overactivation of pro-inflammatory pathways progressing towards overt skin effects. PMID- 19131227 TI - Cardiovascular effects in vitro of aqueous extract of wild strawberry (Fragaria vesca, L.) leaves. AB - In contrast to the strawberry fruits, strawberry leaves as a source of bioactive compounds with potentially beneficial biological effects have been largely overlooked. In this study we examined direct, dose-dependent effects of wild strawberry (Fragaria vesca, L.) leaves aqueous extract, in two experimental models and animal species, the isolated guinea pig hearts and rat aortic rings. Vasodilatory potential of the wild strawberry leaves extract was compared with vasodilatory activity of aqueous extract of hawthorn (Crataegus oxycantha, L) leaves with flowers, which can be regarded as a reference plant extract with a marked vasodilatory activity. The extracts were analysed by their "phenolic fingerprints", total phenolic content and antioxidative capacity. Their vasodilatory activity was determined and compared in the isolated aortic rings from 24 rats that were exposed to the extracts doses of 0.06, 0.6, 6, and 60 mg/100ml. Both extracts induced similar, dose-dependent vasodilation. Maximal relaxation was 72.2+/-4.4% and 81.3+/-4.5%, induced by the strawberry and hawthorn extract, respectively. To determine vasodilatory mechanisms of the wild strawberry leaves extract, endothelium-denuded and intact rings exposed to nitric oxide (NO) synthase inhibitor L-NAME or cyclooxygenase inhibitor indomethacin were used. Removal of the endothelium prevented and exposure to L-NAME or indomethacin strongly diminished the vasodilatatory response to the extract. In the isolated hearts (n=12), the wild strawberry extract was applied at concentrations of 0.06, 0.18, 0.6, and 1.8 mg/100ml. Each dose was perfused for 3.5 min with 15 min of washout periods. Heart contractility, electrophysiological activity, coronary flow and oxygen consumption were continuously monitored. The extract did not significantly affect heart rate and contractility, main parameters of the cardiac action that determine oxygen demands, while coronary flow increased up to 45% over control value with a simultaneous decrease of oxygen extraction by 34%. The results indicate that the aqueous extract of wild strawberry leaves is a direct, endothelium-dependent vasodilator, action of which is mediated by NO and cyclooxygenase products and which potency is similar to that of the hawthorn aqueous extract. PMID- 19131228 TI - Bimodal effect of humic acids on the LPS-induced TNF-alpha release from differentiated U937 cells. AB - Humic substances (HS) have been reported to possess anti-inflammatory as well as pro-inflammatory properties. The anti-inflammatory activity was demonstrated in the rat paw edema model and we found a preliminary explanation in the 5 lipoxygenase inhibitory effect of humic acids (HA). The pro-inflammatory activity is reflected by the production and release of pro-inflammatory cytokines in HA treated neutrophilic granulocytes. With regard to the potential use of HA as antiviral and UV-protective agents it appears advisable to investigate the role of HS in the inflammation process in more detail. Hence we tested four different HS preparations - two naturally occurring HA from the Altteich peatland in Germany, one fulvic acid (FA) preparation from a Finnish spruce forest and a synthetic HA-like polymer (caffeic acid oxidation product, KOP) for their influence on the lipopolysaccharide (LPS)-induced TNF-alpha release in human U937 cells. In addition, the cytotoxicity of HS was determined. The results demonstrate a concentration-dependent bimodal effect of HA on the TNF-alpha release of differentiated LPS-stimulated U937 cells for both the natural black peat HA from the Altteich peatland and the HA-like polymer KOP. Low HA concentrations (10-80 microg/ml) enhanced the TNF-alpha release by up to threefold (pro-inflammatory activity), while HA concentrations >100 microg/ml reduced it about 10-fold (anti-inflammatory activity). FA failed to enhance TNF alpha release, but reduced it at higher concentrations (>200 microg/ml) by the half. Brown water HA did not exert any significant effect on TNF-alpha release. No HS-stimulated TNF-alpha release was also observed in the absence of exogenously supplied LPS. This means that HS, unlike endotoxin, are no inflammation-causing agents for LPS-untreated cells. Differences in the effect of individual HS on TNF-alpha release are discussed in connection with the polyanionic character of HS, their molecular mass distribution and the hitherto imperfectly known chemical structure. PMID- 19131229 TI - Extract of Ginkgo biloba induces glutathione-S-transferase subunit-P1 in vitro. AB - The extract of Ginkgo biloba (EGb), containing 24% flavone glycosides and 6% terpenoids, is widely used to treat early-stage Alzheimer's disease, vascular dementia, peripheral claudication and vascular tinnitus. Its remarkable antioxidant activity has recently been demonstrated in both cell lines and animals. Glutathione-S-transferases (GSTs) are a class of important detoxification enzymes in the antioxidant system and GST-P1 is the major GST isoform highly expressed in human tissues. Over expression of GST-P1 protected prostate cells from cytotoxicity and DNA damage by the heterocyclic amine carcinogen, while inhibition of expression of GST-P1 by transfecting GST-P1 antisense cDNA or targeted deletion of GST-P1 has been found to sensitize cells to cytotoxic chemicals. It is obvious that induction of GST-P1 expression should be a promising alternative for chemoprevention. The present study aimed to investigate the induction effect of EGb on GST-P1 in HepG2 and Hep1c1c7 cell lines and found that GST-P1 was increased both at the expression and enzyme activity levels. PMID- 19131230 TI - Can MTT be used to quantify the antioxidant activity of plant extracts? AB - Antioxidant properties of three different medicinal plant extracts from northern Nigeria were evaluated using the traditional qualitative 2,2, diphenyl picrylhydrazyl (DPPH) method and a new 3-(4,5-dimethylthiazole-2-yl)-2,5 diphenyltetrazolium bromide (MTT) microdilution method described here. The results were in agreement. It is a simple, rapid and inexpensive method compared with many other methods for determining quantitative antioxidant activity of plant extracts. PMID- 19131231 TI - Severe exacerbations and BODE index: two independent risk factors for death in male COPD patients. AB - OBJECTIVES: 1) To determine whether severe exacerbation of COPD is a BODE index independent risk factor for death; 2) whether the combined application of exacerbations and BODE (e-BODE index), offers greater predictive capacity than BODE alone or can simplify the model, by replacing the exercise capacity (BODEx index). METHODS: A prospective study was made of a cohort of COPD patients. In addition to calculation of the BODE index we register frequency of exacerbations. An analysis was made of all-cause mortality, evaluating the predictive capacity of the exacerbations after adjusting for the BODE. These variables were also used to construct two new indexes: e-BODE and BODEx. RESULTS: The study included 185 patients with a mean age of 71+/-9 years, and FEV(1)% 47+/-17%. Severe exacerbation appeared as an independent adverse prognostic variable of BODE index. For each new exacerbation the adjusted mortality risk increased 1.14-fold (95% CI: 1.04-1.25). However, the e-BODE index (C statistic: 0.77, 95% CI: 0.67 0.86) didn't improve prognostic capacity of BODE index (C statistic: 0.75, 95% CI: 0.66-0.84) (p=NS). An interesting finding was that BODEx index (C statistic: 0.74, 95% CI: 0.65-0.83) had similar prognostic capacity than BODE index. CONCLUSIONS: Severe exacerbations of COPD imply an increased mortality risk that is independent of baseline severity of the disease as measured by the BODE index. The combined application of both parameters (e-BODE index) didn't improve the predictive capacity, but on replacing exacerbation with exercise capacity the multidimensional grading system is simplified without loss of predictive capacity. PMID- 19131232 TI - Pompe disease: a neuromuscular disease with respiratory muscle involvement. AB - Pompe disease is a single disease continuum that includes variable neuromuscular symptoms and rates of progression. However, specific clinical features, such as an early onset of respiratory problems preceding limb muscular weakness, distinguish Pompe disease from other neuromuscular diseases in which respiratory insufficiency occurs after loss of ambulation. The management of Pompe disease also differs from other neuromuscular diseases in that specific treatment is now available, making early recognition of the disease a priority. The results from clinical trials with recombinant human acid alpha-glucosidase have been published, and they show promising results with regards to the improvement of respiratory function in patients with Pompe disease. This review aims to give an overview of Pompe disease and to describe the current concepts of the disease. A focus is placed on the pathophysiology and clinical presentation of respiratory muscle involvement in adults. Additionally, new approaches and therapies available for the management of respiratory complications observed in Pompe disease are discussed in detail. PMID- 19131233 TI - Effects of compost biocovers on gas flow and methane oxidation in a landfill cover. AB - Previous publications described the performance of biocovers constructed with a compost layer placed on select areas of a landfill surface characterized by high emissions from March 2004 to April 2005. The biocovers reduced CH(4) emissions 10 fold by hydration of underlying clay soils, thus reducing the overall amount of CH(4) entering them from below, and by oxidation of a greater portion of that CH(4). This paper examines in detail the field observations made on a control cell and a biocover cell from January 1, 2005 to December 31, 2005. Field observations were coupled to a numerical model to contrast the transport and attenuation of CH(4) emissions from these two cells. The model partitioned the biocover's attenuation of CH(4) emission into blockage of landfill gas flow from the underlying waste and from biological oxidation of CH(4). Model inputs were daily water content and temperature collected at different depths using thermocouples and calibrated TDR probes. Simulations of CH(4) transport through the two soil columns depicted lower CH(4) emissions from the biocover relative to the control. Simulated CH(4) emissions averaged 0.0gm(-2)d(-1) in the biocover and 10.25gm(-2)d(-1) in the control, while measured values averaged 0.04gm(-2)d( 1) in the biocover and 14gm(-2)d(-1) in the control. The simulated influx of CH(4) into the biocover (2.7gm(-2)d(-1)) was lower than the simulated value passing into the control cell (29.4gm(-2)d(-1)), confirming that lower emissions from the biocover were caused by blockage of the gas stream. The simulated average rate of biological oxidation predicted by the model was 19.2gm(-2)d(-1) for the control cell as compared to 2.7gm(-2)d(-1) biocover. Even though its V(max) was significantly greater, the biocover oxidized less CH(4) than the control cell because less CH(4) was supplied to it. PMID- 19131234 TI - Leachability of automotive shredder residues burned in a fluidized bed system. AB - This paper presents the results of the study of a combustible fraction of automotive shredder residues (CASRs) and the corresponding ashes generated by combustion on a fluidized bed pilot with the aim to understand the influence of thermal treatment regarding properties for final disposal, such as landfilling. The chemical composition was evaluated and the leachability behaviour of ashes and CASR was investigated using the three more commonly used tests: the European Standard EN 12457, the US TCLP-EPA 1311 and the Dutch availability test EA NEN 7371. Different results were obtained depending on the specific conditions of the methods employed. It was found that both the CASR and the ashes contained large amounts of toxic metals and other undesirable elements, such as Cl and S. For the CASR, in addition to the leachability of organic matter above the limit set for hazardous materials, the release of heavy metals, either under alkaline and acidic conditions was significant, revealing the serious risks associated to the landfilling practices still being undertaken worldwide. Release of organic matter from ashes was insignificant, but solubility of sulphates increased and chlorides exceed the hazardous limits in the case of fly ashes. Toxic metals were found to leach from the ashes only under acidic conditions, except Pb and Cu which also leached from finer ashes at alkaline pH. Cr also leached from ashes at alkaline pH values. Both the Dutch availability and TCLP revealed much higher leaching intensities than the European Standard due to the acidity of leachants. However, it was found that ashes may be more resistant to acidification because they exhibit much higher acid neutralization capacity (ANC) than the untreated CASR. The study undertaken shows that thermal valorisation of the combustible fraction of ASR may avoid the risks associated with their landfilling; however, care has to be taken with the ashes because they also behave as hazardous residues. Although, the mass reduction provided by thermal treatment may make landfilling less expensive, a more profitable reutilization of the ashes should be developed. PMID- 19131235 TI - Disposal of agro-industrial by-products by organic cultivation of the culinary and medicinal mushroom Hypsizygus marmoreus. AB - Organic mushroom cultivation is one of the fastest growing segments of agriculture. At the core of the organic philosophy lies a ban on the use of synthetic fertilizers, pesticides and herbicides, in addition to such tenets as animal welfare, energy efficiency, and social justice. Hypsizygus marmoreus (HM) is a highly praised cultivated culinary and medicinal mushroom. The objective of this paper was to assess the suitability of different spawn media and then the potential of various cultivation substrates to support HM mushroom production compatible with organic standards. This objective was met through the setup of a low-cost cultivation infrastructure. First, seven types of spawn media were tested; then we tested 24 substrates made from organic by-products for their biological efficiency (BE) with strain HM 830, using the liquid inoculation method. The best substrate in terms of BE was corn cob with bran and olive press cake, with a BE of 85.6%. The BE of the same composition but without olive press cake was only 67.5%. The next best substrates were cotton straw combinations with a BE of 31.5-53%. The spent mushroom substrate provides a good method for the disposal of solid waste. The guidance provided in this research complies with organic mushroom cultivation standards and can be used to produce certified organic mushrooms. In addition, it allows responsible and beneficial disposal of a large amount of solid agro-industrial waste. PMID- 19131236 TI - Use of waste gypsum to replace natural gypsum as set retarders in portland cement. AB - The present study is focused on clarifying the influence of waste gypsum (WG) in replacing natural gypsum (NG) in the production of ordinary Portland cement (OPC). WG taken from slip casting moulds in a ceramic factory was formed from the hydration of plaster of paris. Clinker and 3-5wt% of WG was ground in a laboratory ball mill to produce cement waste gypsum (CMWG). The same procedure was repeated with NG to substitute WG to prepare cement natural gypsum (CMNG). The properties of NG and WG were investigated via X-ray Diffraction (XRD), X-ray fluorescence (XRF) and differential scanning calorimetry (DSC)/thermogravimetric (TG) to evaluate the properties of CMNG and CMWG. The mechanical properties of cement were tested in terms of setting time, flexural and compressive strength. The XRD result of NG revealed the presence of dihydrate while WG contained dihydrate and hemihydrate. The content of dihydrate and hemihydrates were obtained via DSC/TG, and the results showed that WG and NG contained 12.45% and 1.61% of hemihydrate, respectively. Furthermore, CMWG was found to set faster than CMNG, an average of 15.29% and 13.67% faster for the initial and final setting times, respectively. This was due to the presence of hemihydrate in WG. However, the values obtained for flexural and compressive strength were relatively the same for CMNG and CMWG. Therefore, this result provides evidence that WG can be used as an alternative material to NG in the production of OPC. PMID- 19131237 TI - Quick monitoring of pozzolanic reactivity of waste ashes. AB - This article proposes a quick method of monitoring for pozzolanic reactivity of waste ashes by investigating the electrical conductivity of the suspension at an elevated temperature. This suspension is obtained by mixing tested pozzolan with an ordinary Portland cement (OPC) solution produced by mixing ordinary Portland cement with water. For comparison, silica fume, metakaolin, rice husk ash and river sand - whose pozzolanic reactivities range from reactive to inert - were used in the experimental investigation. The electrical conductivity of the suspension was continually recorded by using an electrical conductivity meter and stored by using a personal computer for a period of slightly over 1day. The indicative parameters that can be related to pozzolanic reactivity were discussed and analyzed in detail. It was found that it is possible to determine the pozzolanic reactivity of fly ash within 28h by using the proposed technique, as compared to 7 or 28 days for the determination of strength activity index according to ASTM. This technique would help concrete technologists to speedily investigate the quality of fly ash for use as a cement replacement in order to alleviate pollution caused by cement production and solve disposal problems of waste ashes. PMID- 19131238 TI - Synergistically improved sensitivity for the detection of specific DNA sequences using polyaniline nanofibers and multi-walled carbon nanotubes composites. AB - A sensitive electrochemical DNA biosensor was successfully realized on polyaniline nanofibers (PANI), multi-walled carbon nanotubes (MWNT) and chitosan (CHIT) modified carbon paste electrode (CPE) based on the synergistic effect between PANI and MWNT nanoparticles in chitosan film. PANI and MWNT nanocomposites resulted in highly enhanced electron conductive and biocompatible nanostructured film, which was examined by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The immobilization of the probe DNA on the surface of electrode was largely improved due to the unique synergistic effect of PANI and MWNT. The DNA hybridization events were monitored with an EIS label-free detection strategy. Under the optimal conditions, the dynamic detection range of this DNA electrochemical biosensor was from 1.0 x 10(-13) to 1.0 x 10(-7)mol/L and a detection limit of 2.7 x 10(-14)mol/L for the detection of DNA specific sequence of the phosphinothricin acetyltransferase gene (PAT, one of the important screening detection genes for the transgenic plants). Simultaneously, the polymerase chain reaction (PCR) amplification of the terminator of nopaline synthase gene (NOS) from the sample of one kind of genetically modified soybean was also detected satisfactorily. PMID- 19131239 TI - Optical modeling of bioluminescence in whole cell biosensors. AB - Bioluminescence-based whole cell biosensors are devices that can be very useful for environmental monitoring applications. The advantages of these devices are that they can be produced as a single-chip, low-power, rugged, inexpensive component, and can be deployed in a variety of non-laboratory settings. However, such biosensors encounter inherent problems in overall system light collection efficiency. The light emitted from the bioluminescent microbial cells is isotropic and passes through various media before it reaches the photon detectors. We studied the bioluminescence distribution and propagation in microbial whole cell biochips. Optical emission and detection were modeled and simulated using an optical ray tracing method. Light emission, transfer and detection were simulated and optimized with respect to two fundamental system parameters: system geometry and bacterial concentration. Optimization elucidated some of the optical aspects of the biochip, e.g. detector radius values between 300 and 750 microm, and bacterial fixation radius values between 800 and 1200 microm. Understanding theses aspects may establish a basis for future optimization of similar chips. PMID- 19131240 TI - The evolution of plant genomes: scaling up from a population perspective. AB - Plant genomes exhibit tremendous diversity in both their size and structure, with genome sizes across land plants ranging over two to three orders of magnitude and significant variation in structural organization was observed across species (EA Kellogg, JL Bennetzen, The evolution of nuclear genome structure in seed plants, Am J Bot 2004, 91:1709-1725). Five plant genomes are now either completely sequenced or in the draft stage; the grape (O Jaillon et al., The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla, Nature 2007, 449:463-467) and papaya (R Ming et al., The draft genome of the transgenic tropical fruit tree papaya (Carica papaya Linnaeus), Nature 2008, 452:991-997) whole genome sequences were reported most recently. Moreover, sequencing of 41 additional genomes is in progress. There is now an emerging consensus that understanding genome evolution requires consideration of the population genetics of genome diversification, and that description of evolutionary forces at the level of populations and within species can help identify the features that led to plant genome diversity (M Lynch, JS Conery, The origins of genome complexity, Science 2003, 302:1401-1404). In this review we focus on advances in our understanding of the mechanisms that drive the diversification of genomes. In particular, we look at the extent to which demographic features such as effective population size changes within species can drive genome evolution, discuss population genetic models of genome diversification associated with transposable element (TE) mobilization, and describe recent studies on the evolution of gene families. PMID- 19131241 TI - Reflections on some pilot trials of gastrin receptor blockade in pancreatic cancer. AB - The experience of synthesising a novel gastrin receptor antagonist gastrazole and taking it into 3 small clinical studies in pancreatic cancer in man is described. The need for such a compound is illustrated by the observation that inhibition of gastric acid secretion by H2 receptor antagonists results in hypergastrinaemia. A large number of cell types have gastrin receptors including pancreatic cancer cells which have been shown to be stimulated by gastrin. Small numbers of pancreatic cancer patients given gastrazole by continuous intravenous infusion showed prolonged survival compared with best supportive care or placebo, and equivalent survival to those given 5 fluouracil. The results suggest a greater benefit for patients with early stage disease. An alternative gastrin receptor antagonist YF 476 is also described which has the advantage of efficacy given by the oral route. This new compound requires to be studied in pancreatic cancer and other diseases associated with hypergastrinaemia. PMID- 19131243 TI - Nick-containing oligonucleotides as human topoisomerase I inhibitors. AB - A series of oligonucleotides with various lengths that contain nick and topoisomerase I-binding sites were designed. The interactions between these oligonucleotides and human topoisomerase I were investigated and the most efficient one among them has displayed IC(50) value of 6.3 nM. Our studies have also demonstrated that the position of the nick as well as the length of the oligonucleotides were crucial factors for the inhibition of this nuclear enzyme. PMID- 19131244 TI - Tetracyclic indole inhibitors of hepatitis C virus NS5B-polymerase. AB - We report the evolutionary path from an open-chain series to conformationally constrained tetracyclic indole inhibitors of HCV NS5B-polymerase, where the C2 aromatic is tethered to the indole nitrogen. SAR studies led to the discovery of zwitterionic compounds endowed with good intrinsic enzyme affinity and cell-based potency, as well as superior DMPK profiles to their acyclic counterparts, and ultimately to the identification of a pre-clinical candidate with an excellent predicted human pharmacokinetic profile. PMID- 19131245 TI - 5-Nitrofuran-2-yl derivatives: synthesis and inhibitory activities against growing and dormant mycobacterium species. AB - Eighteen 5-nitrofuran-2-yl derivatives were prepared by reacting 5-nitro-2 furfural with various (sub)phenyl/pyridyl thiosemicarbazide using microwave irradiation. The compounds were tested for their in vitro activity against tubercular and various non-tubercular mycobacterium species in log-phase and 6 week-starved cultures. Compound N-(3,5-dibromopyridin-2-yl)-2-((5-nitrofuran-2 yl)methylene)hydrazinecarbothioamide (4r) was found to be the most potent compound (MIC: 0.22 microM) and was 3 times more active than standard isoniazid (INH) and equally active as rifampicin (RIF) in log-phase culture of Mycobacterium tuberculosis H37Rv. In starved M. tuberculosis H37Rv, 4r inhibited with MIC of 13.9 microM and was found to be 50 times more active than INH and slightly more active than RIF. PMID- 19131242 TI - The cure of cancer: a European perspective. AB - Cancer survival analyses based on cancer registry data do not provide direct information on the main aim of cancer treatment, the cure of the patient. In fact, classic survival indicators do not distinguish between patients who are cured, and patients who will die of their disease and in whom prolongation of survival is the main objective of treatment. In this study, we applied parametric cure models to the cancer incidence and follow-up data provided by 49 EUROCARE-4 (European Cancer Registry-based study, fourth edition) cancer registries, with the aims of providing additional insights into the survival of European cancer patients diagnosed from 1988 to 1999, and of investigating between-population differences. Between-country estimates the proportion of cured patients varied from about 4-13% for lung cancer, from 9% to 30% for stomach cancer, from 25% to 49% for colon and rectum cancer, and from 55% to 73% for breast cancer. For all cancers combined, estimates varied between 21% and 47% in men, and 38% and 59% in women and were influenced by the distribution of cases by cancer site. Countries with high proportions of cured and long fatal case survival times for all cancers combined were characterised by generally favourable case mix. For the European pool of cases both the proportion of cured and the survival time of fatal cases were associated with age, and increased from the early to the latest diagnosis period. The increases over time in the proportions of Europeans estimated cured of lung, stomach and colon and rectum cancers are noteworthy and suggest genuine progress in cancer control. The proportion of cured of all cancers combined is a useful general indicator of cancer control as it reflects progress in diagnosis and treatment, as well as success in the prevention of rapidly fatal cancers. PMID- 19131246 TI - Development of dual-acting prodrugs for circumventing multidrug resistance. AB - We have developed a novel dual-acting maleimide-bearing prodrug that incorporates the anticancer agent doxorubicin and an inhibitor of the P-glycoprotein efflux pump that is over-expressed in multidrug resistant tumor cells. Additionally, the prodrug contains a 1,6-self-immolative spacer coupled to the dipeptide Phe-Lys that acts as a substrate for cathepsin B. The prodrug, once bound through its maleimide moiety to the cysteine-34 group of human serum albumin, was cleaved by cathepsin B and in tumor homogenates demonstrating a release of the anticancer agent doxorubicin and the inhibitor. PMID- 19131247 TI - Discovery of trisubstituted cyclohexanes as potent CC chemokine receptor 2 (CCR2) antagonists. AB - A series of trisubstituted cyclohexanes was designed, synthesized and evaluated as CC chemokine receptor 2 (CCR2) antagonists. This led to the identification of two distinct substitution patterns about the cyclohexane ring as potent and selective CCR2 antagonists. Compound 36 exhibited excellent binding (CCR2 IC(50)=2.4 nM) and functional antagonism (calcium flux IC(50)=2.0 nM and chemotaxis IC(50)=5.1 nM). PMID- 19131248 TI - N-Benzyl-1-heteroaryl-3-(trifluoromethyl)-1H-pyrazole-5-carboxamides as inhibitors of co-activator associated arginine methyltransferase 1 (CARM1). AB - A series of N-benzyl-1-heteroaryl-3-(trifluoromethyl)-1H-pyrazole-5-carboxamides targeting co-activator associated arginine methyltransferase 1 (CARM1) have been designed and synthesized. The potency of these inhibitors was influenced by the nature of the heteroaryl fragment with the thiophene analogues being superior to thiazole, pyridine, isoindoline and benzofuran based inhibitors. PMID- 19131249 TI - Nurses' perceptions of leadership in an adult intensive care unit: a phenomenology study. AB - The purpose of this research was to explore ICU nurses' perceptions of nursing leadership in the adult intensive care unit (ICU). The nursing profession needs leaders at all levels; ward, administration and executive and in an era in which there is a shortage of ICU nurses, nursing leadership is important, as positive leadership skills correlate with enhanced recruitment and retention of these specialist nurses. Six ICU nurses with at least 5 years experience in ICU nursing were recruited from a metropolitan hospital in Australia. Qualitative phenomenological methodology was used to depict the lived experiences of nurses' leadership in the adult ICU. Data were collected through individual semi structured interviews using open-ended questions and analysed using Giorgi's [Giorgi A. Toward phenomenologically based research in psychology. J Phenomenol Psychol 1970;1:75-98] descriptive method for data analysis. Five themes emerged and these were all inter-related: leading by example, communication, ability to think outside the management square, knowing your staff and stepping up in times of crisis. These findings highlight the importance of nursing leadership in the adult ICU and the need to ensure that all current and future nursing ICU leaders are adequately prepared and educated for this role. This information may also be used to assist in the development of leadership skills in ICU nurses. PMID- 19131250 TI - A comparison of Star Excursion Balance Test reach distances between ACL deficient patients and asymptomatic controls. AB - ACL injury has been associated with a decrease in proprioceptive performance and specifically postural control. Tests of postural control have been criticised for not being sufficiently challenging. The Star Excursion Balance Test (SEBT) has been proposed to offer sufficient challenge to be a sensitive test for detecting performance deficits related to pathology. The purpose of this study was to determine if decrements SEBT reach distance is associated with ACL deficiency (ACLD). Twenty five ACLD patients ACLD (17 male and 8 female, mean age 30 (SD 4.5) years) and twenty five matched controls were examined carrying out the SEBT. Factorial ANOVA showed the main effects of limb (p=0.006) and direction (p<0.001) and interaction of limb and direction (p=0.015) all had significant differences between the groups. Further analysis revealed significant differences between the control group and the ACLD limb for the limb movement directions of anterior (p=0.0032), lateral (p=0.005), posterior-medial (p=0.0024) and medial (p=0.001). There were also significant differences between the control limbs and uninjured limb of the patients for the directions of medial (p=0.001) and lateral (p=0.001). ACLD patients appear to have deficiencies in their dynamic postural control when compared to normal asymptomatic subjects. Interestingly, in the ACLD patients, their uninjured leg show deficits compared to the control in two of the four directions the ACLD leg was deficient, this may be indicative of a postural control deficit in these patients, which may have predisposed to the ACL injury and would warrant further study. PMID- 19131251 TI - Preliminary evaluation of pharmacological properties of some xanthone derivatives. AB - A series of xanthone derivatives were synthesized and examined for electrocardiographic, antiarrhythmic, hypotensive and anticonvulsant activities as well as for alpha(1)- and beta(1)-adrenergic binding affinities. Among the investigated compounds, some of them exhibited significant antiarrhythmic and/or hypotensive activity. The data obtained via receptor binding assay are in agreement with pharmacological results and could explain antiarrhythmic and/or hypotensive activity of the newly synthesized structures. PMID- 19131252 TI - Function-oriented synthesis applied to the anti-botulinum natural product toosendanin. AB - Botulinum neurotoxins (BoNTs) are the etiological agents responsible for botulism, a disease characterized by peripheral neuromuscular blockade and a characteristic flaccid paralysis of humans. The natural product toosendanin is a traditional Chinese medicine which has been reported to have anti-botulinum properties in animal models. To establish what chemical functionalities are necessary for the anti-botulinum properties found within toosendanin, a study was initiated with the goal of using function-oriented synthesis (FOS) as a strategy to begin to unravel toosendanin's powerful anti-botulinum properties. From these studies a new synthetic strategy is put forth allowing access to a 4-acetoxy CD fragment analogue (14) of toosendanin, which was achieved from mesityl oxide and acetylacetone in 14 steps. Animal studies on this fragment are also reported. PMID- 19131253 TI - Synthesis and antitumor activity of novel enediyne-linked pyrrolo[2,1 c][1,4]benzodiazepine hybrids. AB - A series of novel pyrrolo[2,1-c][1,4]benzodiazepine (PBD) hybrids linked with enediyne is described. These compounds were prepared by linking C-8 of DC-81 (1) with an enediyne (10-16) through carbon chain linkers to afford PBD hybrid agents 17-23 in good yields. Most of the hybrids on human cancer cell lines exhibited higher cytotoxicity, and an increase in the sub-G1 population than 1. In a previous article, we have demonstrated that DC-81-indole conjugate agents (3-6) are potent inducers of cell apoptosis in melanoma. In the present article, we investigated whether DC-81-enediyne agents possess more cytotoxicity than 6 on human 293T cells. Our data revealed that treatment of 293T cells with DC-81 enediyne resulted in a significant increase of annexin V binding, caspase-3 degradation, and p53 arrest to identify apoptotic cells than 6. These results suggest that the DC-81-enediyne agents are more efficient in inducing apoptosis than DC-81-indole in 293T cells. PMID- 19131254 TI - Cancer preventive agents. Part 8: Chemopreventive effects of stevioside and related compounds. AB - In a search for potential cancer chemopreventive agents from natural resources, stevioside (1), a sweetener, and six related compounds, including two aglycones steviol (6) and isosteviol (7), were screened in an in vitro assay for inhibitory effects on Epstein-Barr virus early antigen activation. Compounds 1, 6 and 7 showed significant activity in this assay and also exhibited strong inhibitory effects in a two-stage carcinogenesis test using mouse skin induced by 7,12 dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA). The inhibitory effects of these three compounds were greater than that of glycyrrhizin. Furthermore, these three compounds significantly inhibited mouse skin carcinogenesis initiated by peroxynitrite and promoted by TPA. Their activities were comparable to that of curcumin. These results suggested that 1, as well as 6 and 7, could be valuable as chemopreventive agents for chemical carcinogenesis. PMID- 19131255 TI - A novel hypocrellin B derivative designed and synthesized by taking consideration to both drug delivery and biological photodynamic activity. AB - For making hypocrellins clinically applicable for phototherapy to vascular diseases, it is mainly focused onto finding a derivative which can be transported fluently in blood system but without serious loss of the inherent activity of its parents. Based on this consideration, a novel 17-3-amino-1-propane-sulfonic acid HB Schiff-base (NSHB) was designed and synthesized in this work. As expected, NSHB is readily dissolved in phosphate buffered saline (PBS) or any other aqueous solvent in a concentration which is suitable for intravenous injection, while the quite higher partition coefficient (5:1) is beneficial to the affinity to biological targets. Based on EPR measurements, it is proved that the photosensitization activity of NSHB to photo-generate semiquinone anion radicals and superoxide anion radical (O*(2)(-)) is even higher than its parent HB, while the ability to generate singlet oxygen ((1)O(2)) is not seriously reduced. In addition, nearly comparable PDT activity to A549 cells for NSHB and HB confirms that the molecular design is successful and NSHB is readily delivered into target tissues via blood circulation after intravenous injection. Furthermore, the quantum yield of (1)O(2) for NSHB is as 12.5 times as that for HB under red light (600-700 nm), which is beneficial to phototherapy to solid tumors. PMID- 19131256 TI - Half of elderly patients routinely treated for colorectal cancer receive a sub standard treatment. AB - BACKGROUND: Several database studies report a lack of care in elderly patients with colorectal cancer. PURPOSE: To describe the management of elderly patients admitted for colorectal cancer; to identify factors associated with standard management according to recommendations and to study factors influencing the survival. PATIENTS AND METHODS: All consecutive patients over 75 years managed for a colorectal adenocarcinoma in our hospital from 1995 to 2000 and followed until 2006 were retrospectively included. The appropriateness of the management of their disease according to the recommendations available at that time was assessed. Several risk factors in receiving the standard cancer treatment were tested using univariate and then multivariate logistic regression. Risk factors of survival were studied using univariate and then multivariate survival analysis. RESULTS: One hundred and ten patients were included. Median age was 82 years (range: 75-96). A surgical treatment was performed in 96 patients. The median overall survival was 32 (1-108) months. A standard cancer treatment according to recommendations was performed in 53 (48%) patients: adjuvant chemotherapy in 6/23 patients with stage III tumour, palliative chemotherapy in 3/18 patients with stage IV tumour and adjuvant radiotherapy in 4/14 patients who had a rectal tumour resection. Multivariate analysis retains tumour stage I or II (OR=7.6, 95% C.I.=[2.9-19.9], p<0.0001) as the only factor associated with standard treatment and presence of metastasis (HR=3.9, 95% C.I. [1.4-10.8], p=0.005), and Charlson's score >3 (HR=28.9, 95% C.I. [2.5-335.6], p=0.001) as independent risk factors of poor survival. CONCLUSIONS: Fifty two percent of elderly patients have had a sub-standard cancer treatment. The majority had a surgical treatment, but only a few received chemotherapy or radiotherapy. Metastasis, older age and Charlson's comorbidity score are the main prognosis factors of poor survival. PMID- 19131257 TI - Post-event spontaneous intrusive recollections and strength of memory for emotional events in men and women. AB - Spontaneous intrusive recollections (SIRs) follow traumatic events in clinical and non-clinical populations. To determine whether any relationship exists between SIRs and enhanced memory for emotional events, participants viewed emotional or neutral films, had their memory for the films tested two days later, and estimated the number of SIRs they experienced for each film. SIR frequency related positively to memory strength, an effect more pronounced in the emotional condition. These findings represent the first demonstration of a relationship between SIRs occurring after an emotional experience and subsequent memory strength for that experience. The results are consistent with the possibility that emotional arousal leads both to elevated SIR frequency and better memory, and that the covert rehearsal associated with SIRs enhances memory for emotional relative to neutral stimuli. Additional evidence of menstrual cycle influences on SIR incidence in female participants appears to merit consideration in future work. PMID- 19131258 TI - Intracapsular osteochondroma of the humeral head in an adult causing restriction of motion: a case report. PMID- 19131259 TI - Simultaneous detection of porcine circovirus type 2, classical swine fever virus, porcine parvovirus and porcine reproductive and respiratory syndrome virus in pigs by multiplex polymerase chain reaction. AB - A multiplex polymerase chain reaction (PCR) was designed for the simultaneous detection of four viruses involved in reproductive and respiratory failure in pigs: porcine circovirus type 2 (PCV-2), porcine parvovirus (PPV), classical swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses. The sensitivity of the multiplex PCR using purified plasmid constructs containing the specific viral target fragments was 2.58x10(7), 2.64x10(5), 2.66x10(7) and 2.73x10(5) copies for PRRSV, PCV-2, CSFV and PPV, respectively. Using the multiplex PCR, co-infections with these four viruses were identified in 26/76 (34.2%) piglets born from sows with reproductive failure in China. This method is a rapid, sensitive and cost effective diagnostic tool for the routine surveillance of viral infections in pigs. PMID- 19131260 TI - The effect of medial versus lateral rectus muscle surgery on distance-near incomitance. AB - PURPOSE: For strabismus with distance-near incomitance (unequal distance and near deviations), it is customary to select lateral or medial rectus muscle surgery based on whether the deviation is greater at distance or near. The purpose of this study is to examine how this choice between medial and lateral rectus muscle surgery affects distance-near incomitance. METHODS: Preoperative and postoperative distance and near deviations were obtained from a retrospective review of medical records of consecutive patients who underwent bilateral medial or lateral rectus muscle recessions or resections between January 1, 2000, and June 30, 2007. RESULTS: A total of 626 procedures were identified. After exclusions, initial procedures included 267 medial rectus muscle recessions that produced an average of 9% more exoshift at near and 159 lateral rectus muscle recessions that produced an average of 34% more esoshift at distance. A larger preoperative distance-near incomitance is associated with a greater reduction in distance-near incomitance with surgery, irrespective of which muscles undergo surgery. Recessions for consecutive deviations and resections for residual deviations exhibited a similar pattern. CONCLUSIONS: Esotropia tends to be greater at near and exotropia greater at distance fixation. The apparent greater effect of medial rectus muscle surgery on near deviation and lateral rectus muscle surgery on distance deviation is probably an artifact of this difference in preoperative characteristics. Whether a deviation is greater with distance or near fixation may not be a major consideration in choosing between medial or lateral rectus muscle surgery. PMID- 19131261 TI - Retained features of embryonic metabolism in the adult MRL mouse. AB - The MRL mouse is an inbred laboratory strain that was derived by selective breeding in 1960 from the rapidly growing LG/J (Large) strain. MRL mice grow to nearly twice the size of other commonly used mouse strains, display uncommonly robust healing and regeneration properties, and express later onset autoimmune traits similar to Systemic Lupus Erythematosis. The regeneration trait (heal) in the MRL mouse maps to 14-20 quantitative trait loci and the autoimmune traits map to 5-8 loci. In this paper we report the metabolic and biochemical features that characterize the adult MRL mouse and distinguish it from C57BL/6 control animals. We found that adult MRL mice have retained a number of features of embryonic metabolism that are normally lost during development in other strains. These include an emphasis on aerobic glycolytic energy metabolism, increased glutamate oxidation, and a reduced capacity for fatty acid oxidation. MRL tissues, including the heart, liver, and regenerating ear hole margins, showed considerable mitochondrial genetic and physiologic reserve, decreased mitochondrial transmembrane potential (DeltaPsi(m)), decreased reactive oxygen species (ROS), and decreased oxidative phosphorylation, yet increased mitochondrial DNA and protein content. The discovery of embryonic metabolic features led us to look for cells that express markers of embryonic stem cells. We found that the adult MRL mouse has retained populations of cells that express the stem cell markers Nanog, Islet-1, and Sox2. These are present in the heart at baseline and highly induced after myocardial injury. The retention of embryonic features of metabolism in adulthood is rare in mammals. The MRL mouse provides a unique experimental window into the relationship between metabolism, stem cell biology, and regeneration. PMID- 19131262 TI - Invasive group B streptococcal infections in a tertiary care hospital between 1998 and 2007 in Japan. AB - OBJECTIVE: To clarify clinical and microbiological features of invasive group B Streptococcus (GBS) disease in Japan. METHODS: This was a retrospective review for the period 1998-2007 of patients across all age groups in Nishi-Kobe Medical Center. Invasive GBS disease was defined as GBS isolation from a normally sterile site or skin/soft tissues. RESULTS: Six infant and 52 adult cases of invasive infection were identified. Diagnosis was limited to bacteremia and meningitis in infants, but varied widely in adults with skin/soft tissue infections and bacteremia being common. The overall fatality rate was 16%. An approximately 2.8 fold increase was found in the incidence among adult patients from the first to the second 5-year period. The most frequent underlying condition was diabetes, with the majority (18/23) of such patients showing poor control (HbA1c >8.0%). Amputation at the knee, ankle, or toes was performed in six diabetic adults with skin/soft tissue infections. Of the strains serotyped, types Ib and III predominated. All 58 strains were susceptible to penicillin; 2% were resistant to erythromycin and 3% were resistant to clindamycin. CONCLUSION: This is the first epidemiological report describing invasive GBS disease in Japan. A significant increase in adult patients was noted, and mortality and morbidity remain substantial. PMID- 19131263 TI - Impact of prior antibiotic use in culture-negative endocarditis: review of 86 cases from southern Pakistan. AB - BACKGROUND: A large number of patients treated at our hospital for endocarditis have negative cultures. Taking into consideration the fact that many of these patients receive antibiotics prior to referral, we decided to study culture negative endocarditis in Pakistan. METHODS: The medical records of all patients admitted to the Aga Khan University Hospital, Pakistan, for the period from 1988 to 2001, with an underlying diagnosis of infective endocarditis (IE) and negative cultures, were reviewed. RESULTS: Of the 159 patients diagnosed with IE by revised Duke criteria, 86 (54.1%) had persistent negative cultures. More than half of these patients (52%) had received antibiotics before being referred to our center. Patients with culture-negative endocarditis were less likely to be classified as definite endocarditis by revised Duke criteria (p<0.001, 95% CI 0.07-0.3) or to have large vegetations (p=0.021, 95% CI 0.05-0.5), and more likely to have a mitral valve prolapse (p=0.003, 95% CI 1.6-2.3). Definite endocarditis (p=0.042, 95% CI 1.02-7.4), heart failure (p=0.008, 95% CI 1.4 12.7), renal failure (p=0.017, 95% CI 1.16-40.7), embolism (p=0.019, 95% CI 1.2 38.8), and neurological complications (p=0.02, 95% CI 1.16-9.2) were associated with an increased mortality. CONCLUSION: Culture-negative endocarditis is very common among patients with IE in Pakistan. The presentation, laboratory findings, and complications are similar to those for culture-positive endocarditis. It is postulated that previous antibiotic treatment is the most common cause of culture negative endocarditis in our hospital. PMID- 19131264 TI - [Optimizing the role of surgeons assistants during a laparoscopic hysterectomy]. PMID- 19131265 TI - Ultrasound-induced inactivation of gram-negative and gram-positive bacteria in secondary treated municipal wastewater. AB - The effect of 24kHz, high energy ultrasound in the presence and absence of titanium dioxide particles on the destruction of different bacteria groups was studied. Applying a total of 1500W/L for 60min (this corresponds to 5400kJ/L specific nominal energy), the mean destruction of gram-negative bacteria such as total coliforms, faecal coliforms and Pseudomonas spp. was 99.5%, 99.2% and 99.7%, respectively. More recalcitrant to sonolytic inactivation were the gram positive bacteria Clostridium perfringens and faecal streptococci with a mean removal of 66% and 84%, respectively. The presence of 5g/L TiO(2) generally enhanced the destruction of gram-negative bacteria, yielding three to five logs reduction. On the other hand, the relatively weak sonochemical inactivation of gram-positive bacteria was only slightly affected by the presence of solid particles. Inactivation was found to follow first-order kinetics regarding bacteria population and was not affected significantly by the wastewater quality. Ultrasound irradiation at 4000kJ/L specific nominal energy and in the presence of 5g/L TiO(2) achieved less than 10(3) CFU/100mL total coliforms, thus meeting USEPA quality standards for wastewater reuse. PMID- 19131266 TI - A study on the in vitro degradation properties of poly(L-lactic acid)/beta tricalcuim phosphate (PLLA/beta-TCP) scaffold under dynamic loading. AB - The objective of this study was to investigate the effects of dynamic loading on the in vitro degradation properties of poly(L-lactic acid)/beta-tricalcuim phosphate (PLLA/beta-TCP) composite scaffolds. These scaffolds were prepared by a technique, namely solvent self-proliferating/model compressing/particulate leaching, and they were incubated in a customized simulated body fluid (SBF) flow system under dynamic loading conditions for 6 weeks. The bioactivity and the degradation behaviors of the composite scaffolds were systematically investigated through the formation of apatite, the mass and porosity changes, the molecular weight changes of PLLA, the compressive strength changes, etc. Results show a high level of apatite deposition on the scaffolds, suggesting their good bioactivity in the SBF. Changes in mass, porosity, molecular weight and compressive strength of the scaffolds happened more under dynamic loading conditions than that under flow only SBF conditions. Dynamic loading with the investigated frequency promoted the degradation of the scaffolds, but did not markedly deteriorate the mechanical properties of the scaffolds. All the results suggest that the composite scaffolds have great potential to be applied in bone replacements or repairs under the in vivo load-bearing conditions. PMID- 19131267 TI - Finite element analysis of stresses developed in the blood sac of a left ventricular assist device. AB - The goal of this research is to develop a 3D finite element (FE) model of a left ventricular assist device (LVAD) to predict stresses in the blood sac. The hyperelastic stress-strain curves for the segmented poly(ether polyurethane urea) (SPEUU) blood sac were determined in both tension and compression using a servo hydraulic testing system at various strain rates. Over the range of strain rates studied, the sac was not strain rate sensitive, however the material response was different for tension versus compression. The experimental tension and compression properties were used in a FE model that consisted of the pusher plate, blood sac and pump case. A quasi-static analysis was used to allow for nonlinearities due to contact and material deformation. The 3D FE model showed that blood sac stresses are not adversely affected by the location of the inlet and outlet ports of the device and that over the systolic ejection phase of the simulation the prediction of blood sac stresses from the full 3D model and an axisymmetric model are the same. Minimizing stresses in the blood sac will increase the longevity of the blood sac in vivo. PMID- 19131269 TI - Aqueous synthesis and characterization of CdS, CdS:Zn(2+) and CdS:Cu(2+) quantum dots. AB - CdS quantum dots of different sizes approximately 3-5nm were prepared using aqueous solutions of CdSO(4) and Na(2)S(2)O(3) at 303K with thioglycerol as stabilizing agent and catalyst. The broad X-ray diffraction pattern and SAED pattern with bright circular spots and clear lattice fringes in the high resolution TEM image indicate cubic phase of the nanocrystals. Band gap changes from the bulk value of 2.4 to 3.16eV in the nanoparticles indicating quantum size effect. The photoluminescence (PL) spectrum is blue-shifted to 528nm. The PL emission band is blue/red-shifted on doping with Zn(2+)/Cu(2+). In the case of Cu(2+), the PL intensity is found to be maximum at 10wt% doping and almost completely quenched at 15wt% doping. On doping with Zn(2+), the intensity and blue-shift of PL are found to be maximum at 2 and 6wt% doping, respectively. PMID- 19131270 TI - The Yale Guideline Recommendation Corpus: a representative sample of the knowledge content of guidelines. AB - OBJECTIVE: To develop and characterize a large, representative sample of guideline recommendations that can be used to better understand how current recommendations are written and to test the adequacy of guideline models. We refer to this sample as the Yale Guideline Recommendation Corpus (YGRC). METHOD: To develop the YGRC, we extracted recommendations from guidelines downloaded from the National Guideline Clearinghouse (NGC). We evaluated the representativeness of the YGRC by comparing the frequency of use of controlled vocabulary terms in the YGRC sample and in the NGC. We examined semantic and formatting indicators that were used to denote recommendation statements. RESULTS: In the course of reviewing 7527 recommendation statements, we extracted 1275 recommendations from the NGC and characterized the guidelines from which they were derived. Both semantic and formatting indicators were used inconsistently to denote recommendations. Recommendation statements were not reliably identifiable in 31.6% (310/982) of the guidelines and many recommendations were not executable as written. We also found variability and inconsistency in the way strength of recommendation is currently reported. Over half of the recommendations (52.7%), did not indicate strength, while 6.5% inaccurately indicated strength. CONCLUSION: The YGRC provides a representative sample of current guideline recommendations and demonstrates considerable variability and inconsistency in the way recommendations are written and in the way the recommendation strength is currently reported. PMID- 19131271 TI - Genotyping of acute HBV isolates from England, 1997-2001. AB - BACKGROUND: Increasing data shows the relevance of HBV genotypes in the outcome of infection. Most studies investigating the relationship between the genotypic characteristics of hepatitis B virus (HBV) and the clinical or epidemiological aspects of HBV infection originate from studies of patients with chronic rather than acute hepatitis B. OBJECTIVES: To study a convenience sample representing ca. 5% of reported acute hepatitis B in England between 1997 and 2001 to investigate the distribution of HBV genotypes and specific HBV variants with epidemiological risk factors, thereby providing baseline data for ongoing surveillance. STUDY DESIGN: From 160 serum samples, PCR was carried out to amplify the first 600 bases of the HBV S gene. Amplicons were sequenced and subjected to phylogenetic analysis and risk factor analysis. RESULTS: Fifty-seven percent of the study samples carried HBV belonging to subtype A2, 13% to subtype D2, and the rest to genotype E (8%) and subtypes C2 and D3 (each 6%), D1 and D4 (each 3%) and B4 (1%). One particular A2 isolate was dominant, accounting for 23% of the total sample set. Drug use and homosexual transmission were equally implicated as risks within genotype A2. No mutations associated with vaccine escape or resistance to antiviral therapy were identified. CONCLUSION: Immigration and travel likely shape the observed genotype distribution and consequent prevalence of genotypes other than A2 or D in this population. Data suggests no genetic separation of parenteral and sexually transmitted virus. These data demonstrate the value in pursuing more extensive and recent surveillance. PMID- 19131268 TI - Beyond the number domain. AB - In a world without numbers, we would be unable to build a skyscraper, hold a national election, plan a wedding or pay for a chicken at the market. The numerical symbols used in all these behaviors build on the approximate number system (ANS) which represents the number of discrete objects or events as a continuous mental magnitude. Here, we first discuss evidence that the ANS bears a set of behavioral and brain signatures that are universally displayed across animal species, human cultures and development. We then turn to the question of whether the ANS constitutes a specialized cognitive and neural domain - a question central to understanding how this system works, the nature of its evolutionary and developmental trajectory and its physical instantiation in the brain. PMID- 19131272 TI - Detection, genetic characterization, and quantification of norovirus RNA from sera of children with gastroenteritis. AB - BACKGROUND: Norovirus (NoV) infection is thought to be confined to the intestines, whereas many reports suggest antigenemia and viremia occur during rotavirus gastroenteritis. OBJECTIVES: To detect NoV RNA in sera and cerebrospinal fluids (CSF) from NoV-infected children, and to quantify and genetically characterize the NoV found in these compartments. STUDY DESIGN: Semi nested PCR was conducted on stool, serum and CSF samples from 56 patients with acute gastroenteritis. Positive samples for NoV were analyzed further by sequencing and real-time PCR. RESULTS: From 39 patients with NoV RNA in stools, 6 also had NoV RNA in sera and none had NoV RNA in CSF. Genotypes of the NoV in stool and serum from the same patient matched completely. The strains in this study had high homology (98.1-100%) with registered strains in the database. The median viral load in stools of the serum-positive patients was greater than that of the serum-negative patients, but this difference was not statistically significant (9.8 x 10(9)copies/g versus 1.1 x 10(9)copies/g (p=0.117)). CONCLUSIONS: NoV RNA appeared in the blood stream in 15% of the patients of NoV gastroenteritis. Although the viral load in stool was not statistically correlated with NoV appearance in serum, genetic analysis indicated that NoV RNA in sera originated from the NoV gastroenteritis. PMID- 19131273 TI - Serologic detection of herpes simplex virus type 2 antibodies among pregnant women using a point-of-care test from Focus Diagnostics. AB - BACKGROUND: Serologic assays that identify herpes simplex type 2 (HSV-2) type specific antibodies have been commercially available for more than a decade. Greater acceptance of these tests is hindered by uncertainty regarding their performance in real-world clinical settings. OBJECTIVES: The primary objective was to compare the test characteristics of the Focus HerpeSelect Express Assay (EA) versus the Focus HerpeSelect enzyme linked immunoassay (ELISA) for detection of HSV-2 type-specific antibodies among pregnant women enrolled from 3 geographic sites with varying prevalences of HSV-2 infection. A second objective was to evaluate the performance of a HSV-2 testing strategy in which EA screens and ELISA confirms HSV-2 serodiagnosis. STUDY DESIGN: We enrolled 399 pregnant women from Atlanta, GA, Moorestown, NJ, and Pittsburgh, PA into this cross-sectional investigation. Capillary whole blood was obtained from study participants, and evaluated for the presence of type-specific HSV-2 antibodies using the EA. Serum samples were also obtained from all study participants for subsequent identification of HSV-2 type-specific antibodies using both ELISA and the Focus Immunoblot assays. RESULTS: We observed 96.2% agreement between results obtained with EA and ELISA. Overall, when compared to ELISA results, the sensitivity of EA for detection of HSV-2 type-specific antibodies was 94.2% and the specificity was 97.1%. Using Immunoblot results as our standard for performance calculations, the positive predictive value (PPV) of HSV-2 serodiagnosis increased from 91.7% to 98.2% when ELISA was used to confirm EA testing. CONCLUSIONS: EA provides similar results to ELISA for the identification of HSV-2 type-specific antibodies among pregnant women. As use of the point-of-care (POC) EA in conjunction with confirmatory ELISA testing improves the PPV of HSV-2 serodiagnosis compared to the use of EA or ELISA testing alone, validation of this diagnostic algorithm in other at-risk populations may be warranted. PMID- 19131274 TI - Viral etiology and incidence associated with acute gastroenteritis in a 5-year survey in Gyeonggi province, South Korea. AB - BACKGROUND: Acute gastroenteritis caused by viruses is one of the leading causes of childhood morbidity. OBJECTIVE: The incidence and etiology of virus-associated gastroenteritis during 2001-2005 in Gyeonggi province, South Korea was determined. STUDY DESIGN: A total of 10,028 stool specimens were analyzed with enzyme immunoassay, reverse transcription-polymerase chain reaction, and nucleotide sequencing. Phylogenetic analysis of genotype sequences was performed for each of four commonly detected viral pathogens. RESULTS: At least one viral agent was detected in 29.0% (2908 of 10,028) of the stool samples. Among these, rotavirus was most numerous (66.7%) followed by norovirus (12.6%), astrovirus (11.9%), and enteric adenovirus (2.6%). Mixed infections were identified in 180 cases (1.8%). Patients aged 1-5 years had the largest proportion 83.6% (2430 of 2908) of virus-positive stool specimens. Viral-mediated gastroenteritis was most common from December to May. The most prevalent strains of rotavirus, adenovirus, astrovirus, and norovirus were G1 type, type 41, G1 type, and GII-1 type, respectively. CONCLUSIONS: Our finding showed the diversity of viral causative agents of gastroenteritis and the prevalence of each gastroenteritis-associated viral genotype in South Korea. PMID- 19131275 TI - Newborn infants process pitch intervals. AB - OBJECTIVE: We investigated whether the auditory system of newborn babies extracts the constancy of a pitch interval from exemplars varying in absolute pitch. METHODS: Event-related brain potentials (ERP) were recorded from healthy newborn infants in an oddball paradigm consisting of frequent standard and infrequent deviant tone pairs. Tone pairs varied in absolute frequency. Standard and deviant pairs differed in the amount of pitch difference within the pairs, but not in the direction of pitch change. RESULTS: Deviant tone pairs elicited a discriminative ERP response. CONCLUSIONS: This result suggests that the neonate auditory system represents pitch intervals similarly to adults. SIGNIFICANCE: Adult-like processing of pitch intervals allows newborn infants to learn music, speech prosody, and to process various important auditory cues based on spectral acoustic features. PMID- 19131276 TI - The effects of compression garments on recovery of muscle performance following high-intensity sprint and plyometric exercise. AB - This study compared the effects of compression garments on recovery of evoked and voluntary performance following fatiguing exercise. Eleven participants performed 2 sessions separated by 7 days, with and without lower-body compression garments during and 24h post-exercise. Participants performed a 10-min exercise protocol of a 20-m sprint and 10 plyometric bounds every minute. Before, following, 2h and 24h post-exercise, evoked twitch properties of the knee extensors, peak concentric knee extension and flexion force were assessed, with blood samples drawn to measure lactate [La(-)], pH, creatine kinase (CK), aspartate transaminase (AST) and c-reactive protein (C-RP). Heart rate, exertion (RPE) and muscle soreness (MS) measures were obtained pre- and post-exercise. No differences (P=0.50-0.80) and small effect sizes (d<0.3) were present for 20-m sprint (3.59+/-0.22 vs. 3.59+/-0.18s) or bounding performance (17.13+/-1.4 vs. 17.21+/-1.7 m) in garment and control conditions. The decline and recovery in concentric force were not different (P=0.40) between conditions. Full recovery of voluntary performance was observed 2h post-exercise, however, evoked twitch properties remained suppressed 2h post-exercise in both conditions. No differences (P=0.40-0.80, d<0.3) were present between conditions for heart rate, RPE, [La(-)], pH, CK or C-RP. However, 24h post-exercise a smaller change (P=0.08; d=2.5) in AST (23.1+/-3.1 vs. 26.0+/-4.0) and reduced (P=0.01; d=1.1) MS (2.8+/-1.2 vs. 4.5+/-1.4) were present in the garments. In conclusion the effects of compression garments on voluntary performance and recovery were minimal; however, reduced levels of perceived MS were reported following recovery in the garments. PMID- 19131278 TI - Maternal and fetal risk factors associated with severity of hypospadias: a comparison of mild and severe cases. AB - OBJECTIVE: To determine maternal and fetal demographic factors which predict the risk of increasing severity of hypospadias. PATIENTS AND METHODS: A population based study using the Nova Scotia Atlee Perinatal Database was performed. Demographic variables of mothers and boys with hypospadias were obtained from 1980 to 2007 inclusive. Hypospadias was graded by the position of the urinary meatus as glanular, coronal, shaft, or proximal to shaft. Maternal and fetal risk factors for hypospadias severity were compared using logistic regression. RESULTS: The total number of male pregnancy and birth records during the study period was 130,796. The total number of cases of hypospadias was 995, yielding an incidence of 0.76%. The severity of hypospadias was graded as glanular in 428 (77.8%); coronal in 77 (14%); penile shaft in 34 (6.2%); and proximal to the penile shaft in 12 (2.2%). The severity of hypospadias was not graded in 445 cases. Low birth weight, low gestational age and maternal age were associated with increased severity of hypospadias, but only maternal age (P<0.03) when logistic regression was performed. Limitations included self-reporting for some parameters, such as smoking, and lack of data, such as for the use of assisted reproductive technologies. CONCLUSIONS: Advanced maternal age was associated with increased severity of hypospadias in our population. PMID- 19131279 TI - GABA and nitric oxide in the PVN are involved in arterial pressure control but not in the chemoreflex responses in rats. AB - GABAergic, nitrergic and glutamatergic mechanisms in the PVN on the baseline mean arterial pressure (MAP), heart rate (HR) and on the cardiovascular responses to chemoreflex activation in awake rat were evaluated. Chemoreflex was activated with KCN before and after microinjections into the PVN. Bicuculline into the PVN increased baseline MAP (94+/-3 vs 113+/-5 mmHg) and HR (350+/-9 vs 439+/-18 bpm) but had no effect on the pressor (49+/-5 vs 47+/-6 mmHg) or bradicardic (-213+/ 23 vs -256+/-42 bpm) responses (n=7). Kynurenic acid into the PVN (n=6) produced no significant changes in the MAP (98+/-3 vs 100+/-3 mmHg), HR (330+/-5 vs 339+/ 12 mmHg) or in the pressor (50+/-4 vs 42+/-4 mmHg) and bradicardic (-252+/-4 vs 285+/-16 bpm) responses to chemoreflex. L-NAME into the PVN (n=8) produced increase in the MAP (94+/-3 vs 113+/-5 mmHg) and HR (350+/-9 vs 439+/-18 bpm) but had no effect on the pressor (52+/-5 vs 47+/-6 mmHg) or bradicardic (-253+/-19 vs -320+/-25 bpm) responses to chemoreflex. We conclude that GABA(A) and nitric oxide in the PVN are involved in the maintenance of the baseline MAP but not in the modulation of the responses to chemoreflex. The results also show that Glutamate receptors in the PVN are not involved in maintenance of the baseline MAP, HR or in the cardiovascular responses to chemoreflex in awake rats. PMID- 19131280 TI - Algorithms for automatic processing of data from mass spectrometric analyses of lipids. AB - Lipidomics comprises large-scale studies of the structures, quantities, and functions of lipid molecular species. Recently developed mass spectrometric methods for lipid analyses, especially electrospray ionization (ESI) tandem mass spectrometry, permit identification and quantitation of an enormous variety of distinct lipid molecular species from small amounts of biological samples but generate a huge amount of experimental data within a brief interval. Processing such data sets so that comprehensible information is derived from them requires bioinformatics tools, and algorithms developed for proteomics and genomics have provided some strategies that can be directly adapted to lipidomics. The structural diversity and complexity of lipids, however, also requires the development and application of new algorithms and software tools that are specifically directed at processing data from lipid analyses. Several such tools are reviewed here, including LipidQA. This program employs searches of a fragment ion database constructed from acquired and theoretical spectra of a wide variety of lipid molecular species, and raw mass spectrometric data can be processed by the program to achieve identification and quantification of many distinct lipids in mixtures. Other approaches that are reviewed here include LIMSA (Lipid Mass Spectrum Analysis), SECD (Spectrum Extraction from Chromatographic Data), MPIS (Multiple Precursor Ion Scanning), FIDS (Fragment Ion Database Searching), LipidInspector, Lipid Profiler, FAAT (Fatty Acid Analysis Tool), and LIPID Arrays. Internet resources for lipid analyses are also summarized. PMID- 19131281 TI - Determination of cholesterol in food samples using dispersive liquid-liquid microextraction followed by HPLC-UV. AB - A fast, simple, and sensitive sample preparation procedure based on dispersive liquid-liquid microextraction (DLLME) is proposed for the determination of cholesterol in food samples using isocratic reverse phase high performance liquid chromatography (RP-HPLC) and UV detection. The influence of several important parameters on extraction efficiency of cholesterol was evaluated. Under optimized conditions, a linear relationship was obtained between the peak area and the concentration of cholesterol in the range of 0.03-10 microgl(-1). The detection and quantification limits were 0.01 and 0.03 microgl(-1), respectively. Intra-day and inter-day precisions for the analysis of cholesterol were in the range of 1.0 3.1%. The applicability of the proposed method was demonstrated by analyzing cholesterol in milk, egg yolk and olive oil. PMID- 19131282 TI - A combined solid phase extraction/capillary gel electrophoresis method for the determination of phosphorothioate oligodeoxynucleotides in biological fluids, tissues and feces. AB - A novel biological sample clean-up procedure has been developed for the determination of phosphorothioate oligodeoxynucleotides (PS-ODNs) and derived metabolites in biological fluids (plasma, urine and bile) and in tissues and feces from mice and rats. This method uses a one-step C18 solid-phase extraction (SPE) for biological matrix removal, and it uses capillary gel electrophoresis (CGE) for analyte detection. The assay is specific, and its linearity is superb (r>0.99) for IV-AS (a 13-mer PS-ODN) and PS19 (a 19-mer PS-ODN) in a variety of biological matrices. For both IV-AS and PS19, the precision, accuracy and absolute recovery values were found to be <20%, +/-20% and 80-120%, respectively. The LODs of IV-AS and PS19 were 0.6 mg/l for plasma, 0.8 mg/l for rat urine and bile, 6 microg/g for rat tissues, and 10 microg/g for rat feces, with a signal-to noise ratio of 3 (S/N=3). This method has been successfully applied to the analysis and quantitation of PS-ODNs in various biological samples arising from preclinical pharmacokinetic studies. PMID- 19131283 TI - Elevation of serum gamma-glutamyltranspeptidase activity is frequent in chronic hepatitis C, and is associated with insulin resistance. AB - BACKGROUND AND AIMS: Serum gamma-glutamyltranspeptidase level is often increased in patients with chronic hepatitis C, and we aimed to identify factors associated with this phenomenon in patients completely abstinent from alcohol (teetotaller). PATIENTS AND METHODS: 71 teetotaller patients have been identified by personal history, questioning of relatives, CAGE questionnaire administration and unscheduled alcoholemia measurements. RESULTS: 39 patients (55%) had elevated (>50IU/L) gamma-glutamyltranspeptidase level. Body mass index, insulin and C peptide level, insulin resistance, piecemeal necrosis score > or =3, fibrosis score > or =2 and steatosis score > or =1 were significantly higher in these patients than in those (n=32) with normal gamma-glutamyltranspeptidase. At multiple linear regression analysis gamma-glutamyltranspeptidase level was associated with C-peptide level, insulin resistance and histopathologic grading. At multiple logistic regression analysis, C-peptide level (OR=2.13) and piecemeal necrosis score > or =3 (OR=4.59) were the only factors independently associated with elevated gamma-glutamyltranspeptidase. Sustained virological response during pegylated interferon plus ribavirine treatment was achieved by 97% and 49% patients with normal and elevated gamma-glutamyltranspeptidase, respectively (p=0.0001). CONCLUSION: Serum gamma-glutamyltranspeptidase level is often elevated in chronic hepatitis C and is associated with metabolic and inflammatory factors; this phenomenon may contribute to explain and to predict resistance to treatment in this subgroup of patients. PMID- 19131284 TI - Identifying factors contributing to reduced breast tumor size: a longitudinal study. AB - AIM: This study examines the trends and outcomes of breast cancer patients who have undergone surgical procedures at the Department of Surgical Sciences, University of Insubria, Varese, Italy. It also identifies the factors that contributed to the reduction of the breast tumor size over a 13-year period at a tertiary referral center. METHODS: All breast cancer operations performed at the Department of Surgical Sciences, University of Insubria, Varese, Italy, from January 1992 to June 2005 were examined and data from their surgical pathology reports were also analyzed, using a prospective database. A longitudinal study was performed to compare and analyze the pathological data during three consecutive time periods. The periods were from 1992 to 1996, 1997 to 1999, and 2000 to 2005. Surgical and pathological outcomes included age of the patient at the time of the diagnosis, partial breast resections, mastectomies, axillary lymphadenectomies, tumor size, histological type and stage, and lymph node status. RESULTS: The study group was comprised of 3050 patients who underwent breast resection between 1992 and 2005. Quadrantectomy was the preferred surgical approach in 1759 patients (58%). Throughout the longitudinal study, the tumors measuring less than 1cm increased from 13.4% to 15.4%; the number of tumors diagnosed at stage I increased from 44.1% to 56.8%; the most frequent histological type was ductal carcinoma; the number of ductal carcinomas in situ (DCIS) increased from 4% to 6%; and the incidence of lymphadenectomies decreased from 71.6% to 52.5%. Perioperative factors that correlated with the decreased size of the tumor over time were: screening, improvement of diagnostic and therapeutic techniques, and the increased operative use of sentinel lymph node biopsy (SLNB). CONCLUSIONS: There has been an evolving refinement in surgical technique and perioperative management of breast cancer patients undergoing surgical resection at the Department of Surgical Sciences, University of Insubria, Varese, Italy, during the past decades. The present longitudinal study on 3050 surgical breast cancer patients confirmed the progressive reduction of tumor size at the time of the diagnosis. Perioperative factors that correlated with the decreased tumor size over time were mammography screening, improvement of diagnostic and therapeutic techniques, and the use of SLNB. Furthermore, the study showed that the progressive reduced number of useless axillary lymphadenectomies was mainly due to the increased intraoperative use of axillary SLNB. PMID- 19131285 TI - Axillary sentinel lymph node biopsy: an overview. AB - The sentinel lymph node biopsy has become the standard procedure in the staging and management of axillary lymph nodes in early breast cancer. This procedure has been validated by several studies with an accuracy of 95% and a false-negative rate ranging from 5 to 15%. Aim of this study is to analyze the most updated results and open problems reported in literature in the use of sentinel node biopsy in breast surgery. PMID- 19131286 TI - Applying a managerial approach to day surgery. AB - The present article explores the day surgery topic assuming a managerial perspective. If we assume such a perspective, day surgery can be considered as a business model decision care and not just a surgical procedure alternative to the traditional ones requiring patient hospitalization. In this article we highlight the main steps required to develop a strategic approach [Cotta Ramusino E, Onetti A. Strategia d'Impresa. Milano; Il Sole 24 Ore; Second Edition, 2007] at hospital level (Onetti A, Greulich A. Strategic management in hospitals: the balanced scorecard approach. Milano: Giuffe; 2003) and to make day surgery part of it. It means understanding: - how and when day surgery can improve the health care providers' overall performance both in terms of clinical effectiveness and financial results, and, - how to organize and integrate it with the other hospital activities in order to make it work. Approaching day surgery as a business model decision requires to address in advance a list of potential issues and necessitates of continued audit to verify the results. If it does happen, day surgery can be both safe and cost effective and impact positively on surgical patient satisfaction. We propose a sort of "check-up list" useful to hospital managers and doctors that are evaluating the option of introducing day surgery or are trying to optimize it. PMID- 19131287 TI - Breast cancer surgery in an ambulatory setting. AB - AIM: To evaluate the feasibility and efficacy of outpatient surgery for early breast cancer in an Italian ambulatory setting and to assess its benefits. PATIENTS AND METHODS: A review of 88 women treated for breast cancer from an outpatient facility was undertaken from July 2003 to December 2006. The patients were selected for ambulatory surgery according to specific social, environmental, physical and oncological criteria. RESULTS: Eighty-eight women underwent a total of 107 surgical interventions in an ambulatory setting. Sixty out of the eighty eight patients (68%) received a one-day conclusive surgical treatment, and the remaining 28 patients were promptly treated in two phases. Among this latter group, 18 patients (68%) were treated only in an outpatient facility, whereas the other 10 patients require reintervention with hospitalization. There were no intraoperative complications. In the postoperative period, 14 complications were observed: 6 wound infections, 3 hematomas, 1 axillary seroma and 4 readmissions. The patients' readmissions were due to nausea and emesis in one case, disphnoea in another case, and only two readmissions were due to surgical complications (hematoma in both cases). Patients that were interviewed exhibited a high level of satisfaction from the treatments they received. DISCUSSION: This study confirms the feasibility, efficacy and safety of the outpatient setting regime, which is highly appreciated by women and is more cost effective than surgery in a hospital setting. PMID- 19131288 TI - Congenital anomalies in diabetic pregnancy. AB - Congenital malformations are more common in infants of diabetic women than in children of non-diabetic women. The etiology, pathogenesis and prevention of the diabetes-induced malformations have spurred considerable clinical and basic research efforts. The ultimate aim of these studies has been to obtain an understanding of the teratogenic process, which may enable precise preventive therapeutic measures in diabetic pregnancies. The results of the clinical and basic studies support the view of an early gestational induction of the malformations in diabetic pregnancy by a teratogenic process of multifactorial etiology. There may be possible targets for new therapeutic efforts revealed by the research work. Thus, future additions to the therapeutic efforts may include supplementation with antioxidants and/or folic acid, although more research is needed to delineate the dosages and compounds to be used. As the research into genetic predisposition for the teratogenic induction of malformations by maternal diabetes starts to reveal new genes and gene products involved in the etiology of the malformations, a set of new targets for intervention may arise. PMID- 19131289 TI - The transverse myocutaneous gracilis flap: technical refinements. AB - Autologous free tissue transfer is an ideal method for breast reconstruction. The deep inferior epigastric perforator (DIEP) flap is considered the gold-standard procedure worldwide. However, in selected patients this flap cannot be performed to achieve satisfactory outcomes. The transverse myocutaneous gracilis (TMG) flap is one of the most recent additions to the armamentarium of breast-reconstructive surgeons. This flap can provide adequate autologous tissue with a hidden scar. Since its description for breast reconstruction in 2004, no series have been published and its recognition is still lacking. The main criticism of this flap is the lack of volume that can be achieved and the potential for donor morbidity. We report upon a 2-year experience with the use of TMG flaps for breast reconstruction, assessing the potential indications and introducing some technical refinements in order to expand the role of this flap in breast reconstruction. MATERIALS AND METHODS: Information regarding all TMG flaps performed in the period between January 2006 and December 2007 was prospectively collected. Indications and outcomes were reviewed. The surgical technique was revised and standardised to achieve a routine set-up. RESULTS: During the study period, 19 TMG flaps were performed in 12 patients (seven double procedures: five bilateral cases and two stacked flaps for unilateral breast reconstruction). One flap was lost 9 days postoperatively. Follow-up ranged from 6 months to 2 years. We detail our surgical technique and describe refinements to speed up flap harvest, increase flap volume, optimise flap inset and minimise donor-site complications. CONCLUSION: Although the DIEP flap is still our preferred choice for breast reconstruction, the TMG flap is suitable as a first-line option in small-to-moderate breasted women or as a second-line choice for larger-breasted women for whom the DIEP flap may not be the preferred choice. It is also a reliable salvage flap in cases of previous flap failure. PMID- 19131291 TI - Therapeutic applications of the larvae for wound debridement. AB - It has been known for centuries that application of larvae is useful to heal certain wounds by facilitating debridement of necrotic tissue. Their therapeutic use was popularised in the beginning of the 19th century, but waned in the 1940s with the advent of antiseptic wound management and antibiotics. In more recent years, larvae are once again in vogue for management of difficult wounds. The mechanism of wound debridement by larvae includes the complete wound by continuous larval motion, secretion of proteolytic enzymes and antibacterial substances, effects on epidermal growth factor and interleukin-6 (IL-6) and ingestion and digestion of bacteria and necrotic tissue. In our study, wound debridement was achieved satisfactorily in 29 of 34 patients (85%) with chronic wounds. In the remaining five patients, failures occurred due to inadequate sealing in two patients (6%), death of larvae in two patients (6%) and treatment intolerance in one patient (3%). Larval therapy should be considered as a therapeutic option in the management of certain difficult wounds. PMID- 19131290 TI - Management of split skin graft donor sites--results of a national survey. AB - The authors wished to obtain a 'snapshot' of the range of practice in the management of split skin graft donor sites in the British Isles. MATERIAL/METHODS: Questionnaires were sent to all British consultants and locum consultant plastic surgeons on 1 July 2006. Of the 357 questionnaires, 279 were returned (a response rate of 78%). RESULTS: Alginates were the most popular dressings, especially in adult donor sites--first choice for 167 respondents (60%). Adhesive fabrics were less popular--first choice for small adult donor areas for 46 respondents (16%). Plastic film dressings and Biobrane were even less popular--being the first choice for small and large donor areas, respectively, in children (for approximately 5% of respondents). Ten percent of respondents said they avoid paraffin gauze and another 10% avoid plastic film dressings in all cases. Five percent avoid hydrocolloid and another 5% avoid adhesive fabric in all cases. CONCLUSION: On the basis of these results, we feel that any future study of donor-site dressings should incorporate the most commonly used dressing (alginate) as a control. PMID- 19131293 TI - Dynamic 2D ultrasound and 3D CT image registration of the beating heart. AB - Two-dimensional ultrasound (US) is widely used in minimally invasive cardiac procedures due to its convenience of use and noninvasive nature. However, the low quality of US images often limits their utility as a means for guiding procedures, since it is often difficult to relate the images to their anatomical context. To improve the interpretability of the US images while maintaining US as a flexible anatomical and functional real-time imaging modality, we describe a multimodality image navigation system that integrates 2D US images with their 3D context by registering them to high quality preoperative models based on magnetic resonance imaging (MRI) or computed tomography (CT) images. The mapping from such a model to the patient is completed using spatial and temporal registrations. Spatial registration is performed by a two-step rapid registration method that first approximately aligns the two images as a starting point to an automatic registration procedure. Temporal alignment is performed with the aid of electrocardiograph (ECG) signals and a latency compensation method. Registration accuracy is measured by calculating the TRE. Results show that the error between the US and preoperative images of a beating heart phantom is 1.7 +/-0.4 mm, with a similar performance being observed in in vivo animal experiments. PMID- 19131292 TI - A 3-D high-frequency array based 16 channel photoacoustic microscopy system for in vivo micro-vascular imaging. AB - This paper discusses the design of a novel photoacoustic microscopy imaging system with promise for studying the structure of tissue microvasculature for applications in visualizing angiogenesis. A new 16 channel analog and digital high-frequency array based photoacoustic microscopy system (PAM) was developed using an Nd:YLF pumped tunable dye laser, a 30 MHz piezo composite linear array transducer, and a custom multichannel receiver electronics system. Using offline delay and sum beamforming and beamsteering, phantom images were obtained from a 6 mum carbon fiber in water at a depth of 8 mm. The measured -6 dB lateral and axial spatial resolution of the system was 100+/-5 microm and 45+/-5 microm, respectively. The dynamic focusing capability of the system was demonstrated by imaging a composite carbon fiber matrix through a 12.5 mm imaging depth. Next, 2 D in vivo images were formed of vessels around 100 mum in diameter in the human hand. Three-dimensional in vivo images were also formed of micro-vessels 3 mm below the surface of the skin in two Sprague Dawley rats. PMID- 19131294 TI - Directional view interpolation for compensation of sparse angular sampling in cone-beam CT. AB - In flat detector cone-beam computed tomography and related applications, sparse angular sampling frequently leads to characteristic streak artifacts. To overcome this problem, it has been suggested to generate additional views by means of interpolation. The practicality of this approach is investigated in combination with a dedicated method for angular interpolation of 3-D sinogram data. For this purpose, a novel dedicated shape-driven directional interpolation algorithm based on a structure tensor approach is developed. Quantitative evaluation shows that this method clearly outperforms conventional scene-based interpolation schemes. Furthermore, the image quality trade-offs associated with the use of interpolated intermediate views are systematically evaluated for simulated and clinical cone beam computed tomography data sets of the human head. It is found that utilization of directionally interpolated views significantly reduces streak artifacts and noise, at the expense of small introduced image blur. PMID- 19131295 TI - A unified framework for automated 3-d segmentation of surface-stained living cells and a comprehensive segmentation evaluation. AB - This work presents a unified framework for whole cell segmentation of surface stained living cells from 3-D data sets of fluorescent images. Every step of the process is described, image acquisition, prefiltering, ridge enhancement, cell segmentation, and a segmentation evaluation. The segmentation results from two different automated approaches for segmentation are compared to manual segmentation of the same data using a rigorous evaluation scheme. This revealed that combination of the respective cell types with the most suitable microscopy method resulted in high success rates up to 97%. The described approach permits to automatically perform a statistical analysis of various parameters from living cells. PMID- 19131296 TI - Deformable 2D-3D registration of vascular structures in a one view scenario. AB - Alignment of angiographic 3D scans to 2D projections is an important issue for 3D depth perception and navigation during interventions. Currently, in a setting where only one 2D projection is available, methods employing a rigid transformation model present the state of the art for this problem. In this work, we introduce a method capable of deformably registering 3D vessel structures to a respective single projection of the scene. Our approach addresses the inherent ill-posedness of the problem by incorporating a priori knowledge about the vessel structures into the formulation. We minimize the distance between the 2D points and corresponding projected 3D points together with regularization terms encoding the properties of length preservation of vessel structures and smoothness of deformation. We demonstrate the performance and accuracy of the proposed method by quantitative tests on synthetic examples as well as real angiographic scenes. PMID- 19131297 TI - Noninvasive carotid strain imaging using angular compounding at large beam steered angles: validation in vessel phantoms. AB - Stroke and myocardial infarction are initiated by rupturing vulnerable atherosclerotic plaques. With noninvasive ultrasound elastography, these plaques might be detected in carotid arteries. However, since the ultrasound beam is generally not aligned with the radial direction in which the artery pulsates, radial and circumferential strains need to be derived from axial and lateral data. Conventional techniques to perform this conversion have the disadvantage that lateral strain is required. Since the lateral strain has relatively poor accuracy, the quality of the radial and circumferential strains is reduced. In this study, the radial and circumferential strain estimates are improved by combining axial strain data acquired at multiple insonification angles. Adaptive techniques to correct for grating lobe interference and other artifacts that occur when performing beam steering at large angles are introduced. Acquisitions at multiple angles are performed with a beam steered linear array. For each beam steered angle, there are two spatially restricted regions of the circular vessel cross section where the axial strain is closely aligned with the radial strain and two spatially restricted regions (different from the radial strain regions) where the axial strain is closely aligned with the circumferential strain. These segments with high quality strain estimates are compounded to form radial or circumferential strain images. Compound radial and circumferential strain images were constructed for a homogeneous vessel phantom with a concentric lumen subjected to different intraluminal pressures. Comparison of the elastographic signal-to-noise ratio (SNR(e)) and contrast-to-noise ratio (CNR(e)) revealed that compounding increases the image quality considerably compared to images from 0 degrees information only. SNR(e) and CNR(e) increase up to 2.7 and 6.6 dB, respectively. The highest image quality was achieved by projecting axial data, completed with a small segment determined by either principal component analysis or by application of a rotation matrix. PMID- 19131298 TI - Multi-atlas-based segmentation with local decision fusion--application to cardiac and aortic segmentation in CT scans. AB - A novel atlas-based segmentation approach based on the combination of multiple registrations is presented. Multiple atlases are registered to a target image. To obtain a segmentation of the target, labels of the atlas images are propagated to it. The propagated labels are combined by spatially varying decision fusion weights. These weights are derived from local assessment of the registration success. Furthermore, an atlas selection procedure is proposed that is equivalent to sequential forward selection from statistical pattern recognition theory. The proposed method is compared to three existing atlas-based segmentation approaches, namely 1) single atlas-based segmentation, 2) average-shape atlas based segmentation, and 3) multi-atlas-based segmentation with averaging as decision fusion. These methods were tested on the segmentation of the heart and the aorta in computed tomography scans of the thorax. The results show that the proposed method outperforms other methods and yields results very close to those of an independent human observer. Moreover, the additional atlas selection step led to a faster segmentation at a comparable performance. PMID- 19131299 TI - Minimally redundant 2-D array designs for 3-D medical ultrasound imaging. AB - In real-time ultrasonic 3-D imaging, in addition to difficulties in fabricating and interconnecting 2-D transducer arrays with hundreds of elements, there are also challenges in acquiring and processing data from a large number of ultrasound channels. The coarray (spatial convolution of the transmit and receive arrays) can be used to find efficient array designs that capture all of the spatial frequency content (a transmit-receive element combination corresponds to a spatial frequency) with a reduced number of active channels and firing events. Eliminating the redundancies in the transmit-receive element combinations and firing events reduces the overall system complexity and improves the frame rate. Here we explore four reduced redundancy 2-D array configurations for miniature 3 D ultrasonic imaging systems. Our approach is based on 1) coarray design with reduced redundancy using different subsets of linear arrays constituting the 2-D transducer array, and 2) 3-D scanning using fan-beams (narrow in one dimension and broad in the other dimension) generated by the transmit linear arrays. We form the overall array response through coherent summation of the individual responses of each transmit-receive array pairs. We present theoretical and simulated point spread functions of the array configurations along with quantitative comparison in terms of the front-end complexity and image quality. PMID- 19131300 TI - Exact feature extraction using finite rate of innovation principles with an application to image super-resolution. AB - The accurate registration of multiview images is of central importance in many advanced image processing applications. Image super-resolution, for example, is a typical application where the quality of the super-resolved image is degrading as registration errors increase. Popular registration methods are often based on features extracted from the acquired images. The accuracy of the registration is in this case directly related to the number of extracted features and to the precision at which the features are located: images are best registered when many features are found with a good precision. However, in low-resolution images, only a few features can be extracted and often with a poor precision. By taking a sampling perspective, we propose in this paper new methods for extracting features in low-resolution images in order to develop efficient registration techniques. We consider, in particular, the sampling theory of signals with finite rate of innovation and show that some features of interest for registration can be retrieved perfectly in this framework, thus allowing an exact registration. We also demonstrate through simulations that the sampling model which enables the use of finite rate of innovation principles is well suited for modeling the acquisition of images by a camera. Simulations of image registration and image super-resolution of artificially sampled images are first presented, analyzed and compared to traditional techniques. We finally present favorable experimental results of super-resolution of real images acquired by a digital camera available on the market. PMID- 19131301 TI - A proximal iteration for deconvolving Poisson noisy images using sparse representations. AB - We propose an image deconvolution algorithm when the data is contaminated by Poisson noise. The image to restore is assumed to be sparsely represented in a dictionary of waveforms such as the wavelet or curvelet transforms. Our key contributions are as follows. First, we handle the Poisson noise properly by using the Anscombe variance stabilizing transform leading to a nonlinear degradation equation with additive Gaussian noise. Second, the deconvolution problem is formulated as the minimization of a convex functional with a data fidelity term reflecting the noise properties, and a nonsmooth sparsity-promoting penalty over the image representation coefficients (e.g., l(1) -norm). An additional term is also included in the functional to ensure positivity of the restored image. Third, a fast iterative forward-backward splitting algorithm is proposed to solve the minimization problem. We derive existence and uniqueness conditions of the solution, and establish convergence of the iterative algorithm. Finally, a GCV-based model selection procedure is proposed to objectively select the regularization parameter. Experimental results are carried out to show the striking benefits gained from taking into account the Poisson statistics of the noise. These results also suggest that using sparse-domain regularization may be tractable in many deconvolution applications with Poisson noise such as astronomy and microscopy. PMID- 19131302 TI - Optimal spread spectrum watermark embedding via a multistep feasibility formulation. AB - We consider optimal formulations of spread spectrum watermark embedding where the common requirements of watermarking, such as perceptual closeness of the watermarked image to the cover and detectability of the watermark in the presence of noise and compression, are posed as constraints while one metric pertaining to these requirements is optimized. We propose an algorithmic framework for solving these optimal embedding problems via a multistep feasibility approach that combines projections onto convex sets (POCS) based feasibility watermarking with a bisection parameter search for determining the optimum value of the objective function and the optimum watermarked image. The framework is general and can handle optimal watermark embedding problems with convex and quasi-convex formulations of watermark requirements with assured convergence to the global optimum. The proposed scheme is a natural extension of set-theoretic watermark design and provides a link between convex feasibility and optimization formulations for watermark embedding. We demonstrate a number of optimal watermark embeddings in the proposed framework corresponding to maximal robustness to additive noise, maximal robustness to compression, minimal frequency weighted perceptual distortion, and minimal watermark texture visibility. Experimental results demonstrate that the framework is effective in optimizing the desired characteristic while meeting the constraints. The results also highlight both anticipated and unanticipated competition between the common requirements for watermark embedding. PMID- 19131303 TI - Behavioral-based educational intervention directed toward Egyptian agricultural families to protect children from pesticide exposure. AB - Culturally appropriate educational intervention was developed and directed toward agricultural families to improve knowledge and practice of agricultural families in order to protect their children from exposure to pesticides. Parents (n = 297) were randomly assigned into either lecture or video training group. Ability to recall information or improve practices among parents was evaluated in three consecutive sessions (2 weeks and 1 month apart). All participating families were using pesticides inside homes and in the fields. All families were using flying insect killers inside homes (main ingredients include tetramethrin, sumithrin, pyrethrin, piperonyl butoxide) in different forms. Pesticides applied in the fields are organophosphates (57.9%), carbamates (41.4%), and pyrethroids (28.5%). In both lecture and video training groups, knowledge scores of participants improved significantly in the second and third sessions, as compared to the first session. Although practice scores of both groups improved in subsequent sessions, the changes were not significant. Using non-traditional appropriately designed educational interventions that depend more on visual memory is more likely to ensure sustainability of the gained knowledge as well as practice. PMID- 19131304 TI - A longitudinal study of the impact of village health education on environmental sanitation. AB - Health educators working in developing nations commonly encounter administrative constraints and methodological problems which prevent rigorous program evaluation. Using a time-series design, this study followed approximately 600 families in two Guatemalan villages during a four-year period to measure the impact of community health education on seven indicators of change in environmental sanitation. Adoption of new behavior was accompanied by widespread change in village attitudes and perceptions of health benefits. At the level of the family, change in perception was not predictive of change in behavior. A strong relationship between participation in planned educational events and the adoption of sanitation changes suggests that the formative educational strategy was responsible for stimulating change for five of seven indicators. Suggestions are made about the usefulness of the methodology, ways that private behavioral changes may be measured are discussed, and the advantages of the time-series design for health education evaluation and monitoring are noted. PMID- 19131305 TI - The potential of drama and songs as channels for AIDS education in Africa: a report on focus group findings from Ghana. AB - Among those concerned with fighting AIDS worldwide, it is agreed that the best weapon to change people's behavior is education. In this educational effort, all available channels have to be explored. In Ghana, the Health Education Division (HED) of the Ministry of Health set up three drama troupes to carry out AIDS education on a pilot basis. This study evaluates the impact of one of the troupes on AIDS knowledge, attitude and behavior using the focus group research approach. The findings confirm that drama about a well thought out AIDS-related theme does increase knowledge about AIDS and promotes commendable attitudes toward AIDS. It could also lead to changes in sexual behavior. The study also revealed that songs with AIDS-related themes could serve as important cues to "healthful" action in the environment by reminding listeners to adopt safer sexual behaviors. PMID- 19131306 TI - Lessons learned from health sector reform: a four-country comparison. AB - Various reforms have been undertaken to improve the functioning of health systems in developing countries, but there is limited comparative analysis of reform initiatives. This article discusses health sector reform experiences of four developing countries and identifies the lessons learned. The article is based on the review of background papers on Bangladesh, Pakistan, Indonesia, and Tanzania prepared as part of a multi-country study on health sector reform. Findings suggest that decentralization works effectively while implementing primary and secondary health programs. Decentralization of power and authority to local authorities requires strengthening and supporting these units. Along with the public sector, the private sector plays an effective role in institutional and human resources development as well as in improving service delivery. Community participation facilitates recruitment and development of field workers, facility improvement, and service delivery. For providing financial protection to the poor, there is a need to review user fees and develop affordable health insurance with an exemption mechanism. There is no uniform health sector reform approach; therefore, the experiences of other countries will help countries undertake appropriate reforms. Here, it is important to examine the context and determine the reform measures that constitute the best means in terms of equity, efficiency, and sustainability. PMID- 19131307 TI - A transtheoretical review on smoking cessation. AB - The objective of this review is to draw attention to smoking cessation in view of the Transtheoretical Model (TTM). A large body of literature published in the last 15 years was screened by searching the keywords of "Smoking" and "Transtheoretical Model." The Transtheoretical model consists of stages of change and is used to explain addictive behaviors such as smoking. It addresses behavioral change by taking into consideration factors such as planning and decision-making. This article examines this theory and sets forth the processes of change, according to the positive and negative evaluations and strength of the smoking habit. Although the stages of change and the transitions between these stages have not been clearly defined, it still helps in understanding the nature of some addictive behaviors, particularly smoking. PMID- 19131308 TI - Consensus statement: Defining minimal criteria for reporting the systemic inflammatory response to cardiopulmonary bypass. AB - The causal factors of the systemic inflammatory response to cardiopulmonary bypass (CPB) were correctly identified in the early 1990 s: "... activation of complement, coagulation, fibrinolytic, and kallikrein cascades, activation of neutrophils with degranulation and protease enzyme release, oxygen radical production, and the synthesis of various cytokines from mononuclear cells" [Butler 1993]. Why therefore have clinical advances to curb the systemic inflammatory response proven such a disappointment? Part of the problem is that cardiac surgery has never taken intellectual ownership of this issue, borrowing its diagnosis from critical care medicine and failing to define the minimal criteria that should be measured when reporting on the systemic inflammatory response. An evidence based review of the current literature by many of the coauthors on this paper found that the majority of studies on the systemic inflammatory response did not measure a single one of the causal factors listed above - thus hindering our ability to identify mechanisms of causation and identify drug targets [Landis 2008]. A panel of experts convened at the Outcomes XII meeting, Barbados 2008, drafted the present consensus document in order to provide a framework to guide future studies and interdictions of the systemic inflammatory response. Herein, we have recommended: 1) mandatory reporting of minimal CPB and perfusion criteria that may affect outcomes, 2) reporting of a minimal set of causal inflammatory markers linked to adverse sequelae, and 3) reporting of at least one clinical end-point of organ injury, from a list of endpoints and markers of organ injury that balance practicality with clinical meaningfulness. It is our collective belief that this document will serve as a foundation for furthering our understanding of the influence of CPB practice with the systemic inflammatory response by standardizing the reporting of research findings in the peer-reviewed literature. PMID- 19131309 TI - Is school dental screening a political or a scientific intervention? PMID- 19131311 TI - The boy with the golden tooth: a 1593 case report of the first molded gold crown. AB - The case of the Boy with the Golden Tooth, a "miracle" in a remote village in Silesia, in what is today southwestern Poland, was reported extensively in 1593. Here we report that the hoax, perpetrated by someone close to the family and with knowledge of goldsmith techniques, is the first documented case of the use of a molded gold crown. Using period instruments available to goldsmiths and a 0.001" copper sheet, we reproduced, on a plastic pediatric model, what the gold crown could have looked like. PMID- 19131313 TI - Biological mechanisms in palatogenesis and cleft palate. AB - Clefts of the palate are common birth defects requiring extensive treatment. They appear to be caused by multiple genetic and environmental factors during palatogenesis. This may result in local changes in growth factors, extracellular matrix (ECM), and cell adhesion molecules. Several clefting factors have been implicated by studies in mouse models, while some of these have also been confirmed by genetic screening in humans. Here, we discuss several knockout mouse models to examine the role of specific genes in cleft formation. The cleft is ultimately caused by interference with shelf elevation, attachment, or fusion. Shelf elevation is brought about by mesenchymal proliferation and changes in the ECM induced by growth factors such as TGF-betas. Crucial ECM molecules are collagens, proteoglycans, and glycosaminoglycans. Shelf attachment depends on specific differentiation of the epithelium involving TGF-beta3, sonic hedgehog, and WNT signaling, and correct expression of epithelial adhesion molecules such as E-cadherin. The final fusion requires epithelial apoptosis and epithelium-to mesenchyme transformation regulated by TGF-beta and WNT proteins. Other factors may interact with these signaling pathways and contribute to clefting. Normalization of the biological mechanisms regulating palatogenesis in susceptible fetuses is expected to contribute to cleft prevention. PMID- 19131312 TI - The cells that fill the bill: neural crest and the evolution of craniofacial development. AB - Avian embryos, which have been studied scientifically since Aristotle, continue to persevere as invaluable research tools, especially for our understanding of the development and evolution of the craniofacial skeleton. Whether the topic is beak shape in Darwin's finches or signaling interactions that underlie bone and tooth formation, birds offer advantages for craniofacial biology that uniquely complement the strengths of other vertebrate model systems, such as fish, frogs, and mice. Several papers published during the past few years have helped pinpoint molecular and cellular mechanisms that pattern the face and jaws through experiments that could only have been done together with our feathered friends. Ultimately, such knowledge will be essential for devising novel clinical approaches to treat and/or prevent diseases, injuries, and birth defects that affect the human craniofacial skeleton. Here we review recent insights plucked from avians on key developmental processes that generate craniofacial diversity. PMID- 19131314 TI - Analysis of a band 7/MEC-2 family gene of Porphyromonas gingivalis. AB - In vivo-induced antigen technology has previously been used to identify 115 genes induced in Porphyromonas gingivalis W83 during human infection. The aim of this study was to determine if one of these genes, PG1334, was important for the virulence of P. gingivalis. Analysis of plaque samples from persons with periodontitis revealed that PG1334 was expressed in 88.0% of diseased sites, compared with 42.1% of healthy sites, even though P. gingivalis was detected in equal numbers from both sites. A mutant of PG1334 was found to adhere to and to invade better than the parent strain, but did not persist as well in human coronary artery endothelial cells. Additionally, the mutant did not persist as well in a mouse abscess model. This gene appears to be important for the virulence of P. gingivalis, both in vivo and in vitro. PMID- 19131315 TI - Phosphate regulates osteopontin gene transcription. AB - Extracellular inorganic phosphate (ePi) is a key regulator of cementoblast behavior, both in vivo and in vitro, and results in a marked increase in osteopontin expression in vitro. To examine the molecular mechanisms involved in ePi induction of osteopontin gene expression, we transfected a series of osteopontin promoter-luciferase constructs into OCCM-30 cementoblasts. Our results demonstrate that ePi can directly induce osteopontin gene transcription. The region responsive to ePi signaling was localized to a 53-bp region of the promoter between -1454 and -1401 that contains a glucocorticoid response element (GRE). Mutation of the GRE abolished the ePi response, suggesting that glucocorticoid receptor (GR) signaling is required for ePi-mediated transcription. In addition, treatment of cells with the GR antagonist RU-486 (Mifepristone) prevented promoter activation by ePi. The results presented support a model demonstrating that inorganic phosphate regulates OPN gene transcription in cementoblasts through a pathway that requires a functional GR. PMID- 19131316 TI - Circadian rhythm of osteocalcin in the maxillomandibular complex. AB - The human body displays central circadian rhythms of activity. Recent findings suggest that peripheral tissues, such as bone, possess their own circadian clocks. Studies have shown that osteocalcin protein levels oscillate over a 24 hour period, yet the specific skeletal sites involved and its transcriptional profile remain unknown. The current study aimed to test the hypothesis that peripheral circadian mechanisms regulate transcription driven by the osteocalcin promoter. Transgenic mice harboring the human osteocalcin promoter linked to a luciferase reporter gene were used. Mice of both genders and various ages were analyzed non-invasively at sequential times throughout 24-hour periods. Statistical analyses of luminescent signal intensity of osteogenic activity from multiple skeletal sites indicated a periodicity of ~ 24 hrs. The maxillomandibular complex displayed the most robust oscillatory pattern. These findings have implications for dental treatments in orthodontics and maxillofacial surgery, as well as for the mechanisms underlying bone remodeling in the maxillomandibular complex. PMID- 19131317 TI - A novel mutation in the DSPP gene associated with dentinogenesis imperfecta type II. AB - Hereditary dentin defects are divided into dentinogenesis imperfecta and dentin dysplasia. We identified a family segregating severe dentinogenesis imperfecta. The kindred spanned four generations and showed an autosomal-dominant pattern of inheritance. The proband was a child presenting with a severely affected primary dentition, with wide-open pulp chambers and multiple pulp exposures, resembling a DGI type III (DGI-III) pattern. We hypothesized that a mutation in the DSPP gene is responsible for this severe phenotype. Mutational analyses revealed a novel mutation (c.53T>A, p.V18D) near the intron-exon boundary in the third exon of the DSPP gene. We analyzed the effect of the mutation by means of an in vitro splicing assay, which revealed that the mutation did not affect pre-mRNA splicing. Further studies are needed for a better understanding of the nature of the disease and the development of an appropriate treatment strategy. PMID- 19131318 TI - Evidence of linkage in a Hispanic cohort with a Class III dentofacial phenotype. AB - Despite the prevalence of craniofacial disorders, the genetic contribution remains poorly understood. Class III malocclusion represents a specific craniofacial problem that can be handicapping, both functionally and socially. We hypothesized that the Class III phenotype is genetically linked to specific loci that regulate maxillary or mandibular growth. To determine the region linked to the Class III phenotype in four Hispanic families, we performed a genome-wide scan and linkage analysis using 500 microsatellite markers. Pedigree and linkage analyses revealed that the Class III phenotype (primarily maxillary deficiency) segregates in an autosomal-dominant manner, and that 5 loci (1p22.1, 3q26.2, 11q22, 12q13.13, and 12q23) are suggestive of linkage. Candidate genes within the 12q23 region (ZLR=2.93) include IGF1, HOXC, and COL2A1. Chromosome 1 results (ZLR=2.92) were similar to those reported previously in an Asian cohort with mandibular prognathism, suggesting that a common upstream genetic element may be responsible for both mandibular prognathism and maxillary deficiency. PMID- 19131319 TI - The microbiota of acute apical abscesses. AB - As the breadth of bacterial diversity in the oral cavity has been deciphered by molecular studies, several newly identified species/phylotypes have emerged as potential pathogens. We hypothesized that many of these species/phylotypes could also be involved with the etiology of endodontic abscesses. Abscess aspirates from 42 persons were analyzed for the presence of 81 species/phylotypes by means of a reverse-capture checkerboard hybridization assay. Associations between the most frequently detected taxa were calculated. The most prevalent taxa were Fusobacterium nucleatum, Parvimonas micra, and Porphyromonas endodontalis. Other frequently found taxa included Olsenella uli, streptococci, Eikenella corrodens, some as-yet-uncultivated phylotypes (Bacteroidetes clone X083 and Synergistes clone BA121), and newly named species (Prevotella baroniae and Dialister invisus). Several positive bacterial associations were disclosed. Findings not only strengthen the association of many cultivable species with abscesses, but also include some newly named species and uncultivated phylotypes in the set of candidate pathogens associated with this disease. PMID- 19131320 TI - The effects of smoking on dental care utilization and its costs in Japan. AB - Smoking has been established as an important risk factor for periodontal disease and tooth loss. The purpose of this study was a prospective evaluation of the effects of smoking on dental care utilization and its costs, based on data from 5712 males aged 20-59 yrs. Age, dental health behavior, and history of diabetes were adjusted in a multivariate analysis. Current smokers accrued 14% higher dental care costs than never-smokers over a five-year period. This difference in annual dental care costs was mainly attributable to the increased percentage of participants in the 'higher dental care cost' category among current smokers. There was no clear trend identified for the dose-dependent effects of smoking on dental care utilization and its costs. Past smokers incurred lower dental care costs compared with current smokers. Smoking may have played a key role in the increment of dental care utilization and its costs via deterioration in oral conditions. PMID- 19131321 TI - Dentin caries zones: mineral, structure, and properties. AB - Caries Detector staining reveals 4 zones in dentin containing caries lesions, but characteristics of each zone are not well-defined. We therefore investigated the physical and microstructural properties of carious dentin in the 4 different zones to determine important differences revealed by Caries Detector staining. Six arrested dentin caries lesions and 2 normal controls were Caries-Detector stained, each zone (pink, light pink, transparent, apparently normal) being analyzed by atomic force microscopy (AFM) imaging for microstructure, by AFM nano indentation for mechanical properties, and by transverse digital microradiography (TMR) for mineral content. Microstructure changes, and nanomechanical properties and mineral content significantly decreased across zones. Hydrated elastic modulus and mineral content from normal dentin to pink Caries-Detector-stained dentin ranged from 19.5 [10.6-25.3] GPa to 1.6 [0.0-5.0] GPa and from 42.9 [39.8 44.6] vol% to 12.4 [9.1-14.2] vol%, respectively. Even the most demineralized pink zone contained considerable residual mineral. PMID- 19131322 TI - Self-etching increases matrix metalloproteinase expression in the dentin-pulp complex. AB - In adhesive restorations, one major problem is hybrid layer degradation. At present, this deterioration is explained by the activation of the endogenous matrix metalloproteinases (MMPs) present in dentin due to the acidic property of adhesive systems. We hypothesized that self-etching adhesive should also stimulate the expression of MMPs in odontoblasts. In cultured tooth slices, we evaluated the changes in MMP-2 and proMMP-9 expression in the dentin-pulp complex after self-etching adhesive treatment on dentin cavities in immunochemistry and by zymography. The treatment resulted in increased MMP-2 expression in odontoblasts, as shown by immunohistochemistry. Zymography showed increased proMMP-9 and MMP-2 in dentin under self-etching treatment when pulp was present. These results showed that self-etching adhesive stimulates the secretion of MMPs from the dentin-pulp complex and, more precisely, by odontoblasts, suggesting that odontoblasts participate in hybrid layer degradation. PMID- 19131323 TI - Novel F-releasing composite with improved mechanical properties. AB - In recent years, the authors have been developing novel fluoride-releasing dental composites containing ternary zirconium fluoride chelates. The aim of this study was to improve the physical and mechanical properties of these composites by improving the formulation of the monomers and photoinitiators. The hypothesis was that reduction of hydrophilic monomers and improvement of the photoinitiators could reduce water sorption and significantly increase the mechanical properties of the composite. The degree of conversion of the composites containing different compositions of photoinitiators was studied by Fourier transform near-infrared spectroscopy (FT-NIR). Ten experimental composites containing different compositions of ethoxylated bisphenol-A dimethacrylate (EBPADMA), 1,6-hexanediol dimethacrylate (HDDMA), triethylene glycol dimethacrylate (TEGDMA), and 2,2-bis[4 (2-hydroxy-3-methacryloyloxypropoxy) phenyl]-propane (BisGMA) were tested for flexural strength, viscosity, and water sorption. The experimental composite containing 20% synthesized fluoride-releasing monomer, 30% BisGMA, 30% EBPADMA, and 20% HDDMA showed significantly higher fluoride release and recharge, but physical and mechanical properties similar to those of the control composite containing 40% BisGMA, 40% EBPADMA, and 20% HDDMA. PMID- 19131324 TI - Detection of disease outbreaks by the use of oral manifestations. AB - Oral manifestations of diseases caused by bioterrorist agents could be a potential data source for biosurveillance. This study had the objectives of determining the oral manifestations of diseases caused by bioterrorist agents, measuring the prevalence of these manifestations in emergency department reports, and constructing and evaluating a detection algorithm based on them. We developed a software application to detect oral manifestations in free text and identified positive reports over three years of data. The normal frequency in reports for oral manifestations related to anthrax (including buccal ulcers-sore throat) was 7.46%. The frequency for tularemia was 6.91%. For botulism and smallpox, the frequencies were 0.55% and 0.23%. We simulated outbreaks for these bioterrorism diseases and evaluated the performance of our system. The detection algorithm performed better for smallpox and botulism than for anthrax and tularemia. We found that oral manifestations can be a valuable tool for biosurveillance. PMID- 19131326 TI - Large scale localization of protein phosphorylation by use of electron capture dissociation mass spectrometry. AB - We used on-line electron capture dissociation (ECD) for the large scale identification and localization of sites of phosphorylation. Each FT-ICR ECD event was paired with a linear ion trap collision-induced dissociation (CID) event, allowing a direct comparison of the relative merits of ECD and CID for phosphopeptide identification and site localization. Linear ion trap CID was shown to be most efficient for phosphopeptide identification, whereas FT-ICR ECD was superior for localization of sites of phosphorylation. The combination of confident CID and ECD identification and confident CID and ECD localization is particularly valuable in cases where a phosphopeptide is identified just once within a phosphoproteomics experiment. PMID- 19131325 TI - Striatal alterations of secretogranin-1, somatostatin, prodynorphin, and cholecystokinin peptides in an experimental mouse model of Parkinson disease. AB - The principal causative pathology of Parkinson disease is the progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta projecting to the striatum in the brain. The information regarding the expression of neuropeptides in parkinsonism is very limited. Here we have elucidated striatal neuropeptide mechanisms in experimental parkinsonism using the unilateral 6-hydroxydopamine model to degenerate dopamine neurons. A thoroughly controlled sample preparation technique together with a peptidomics approach and targeted neuropeptide sequence collections enabled sensitive detection, identification, and relative quantitation of a great number of endogenous neuropeptides. Previously not recognized alterations in neuropeptide levels were identified in the unilateral lesioned mice with or without subchronic 3,4 dihydroxy-L-phenylalanine administration, the conventional treatment of Parkinson disease. Several of these peptides originated from the same precursor such as secretogranin-1, somatostatin, prodynorphin, and cholecystokinin. Disease-related biotransformation of precursors into individual peptides was observed in the experimental model of Parkinson disease. Several previously unreported potentially biologically active peptides were also identified from the striatal samples. This study provides further evidence that neuropeptides take part in mediating the central nervous system failure associated with Parkinson disease. PMID- 19131327 TI - A human proteome detection and quantitation project. AB - The lack of sensitive, specific, multiplexable assays for most human proteins is the major technical barrier impeding development of candidate biomarkers into clinically useful tests. Recent progress in mass spectrometry-based assays for proteotypic peptides, particularly those with specific affinity peptide enrichment, offers a systematic and economical path to comprehensive quantitative coverage of the human proteome. A complete suite of assays, e.g. two peptides from the protein product of each of the approximately 20,500 human genes (here termed the human Proteome Detection and Quantitation project), would enable rapid and systematic verification of candidate biomarkers and lay a quantitative foundation for subsequent efforts to define the larger universe of splice variants, post-translational modifications, protein-protein interactions, and tissue localization. PMID- 19131328 TI - Erythrocytic casein kinase II regulates cytoadherence of Plasmodium falciparum infected red blood cells. AB - Plasmodium falciparum malaria is a major human health scourge and a key cause of mortality. Its pathogenicity partly results from the phenomenon of "cytoadherence" mediated by the PfEMP1 (Plasmodium falciparum erythrocyte membrane protein 1) family. Extracellular domains of PfEMP1s are variable and bind various host endothelial receptors, whereas their cytoplasmic domains (VARCs) are relatively conserved. VARCs affix PfEMP1s in the human erythrocyte membrane by interacting with host cytoskeleton proteins and exported parasite proteins. Here, we provide in vitro and in vivo evidence for PfEMP1 phosphorylation (on VARC) and propose an important function for this modification. Specific inhibitors and enhancers have been used to identify erythrocytic casein kinase II (CKII) as the enzyme responsible for VARC modification activity. We have also delineated probable CKII target residues on VARC, which mainly reside in an N-terminal acidic cluster. Our data show that VARC phosphorylation alters its binding to parasite encoded knob-associated histidine-rich protein (KAHRP). Finally, we demonstrate reduced cytoadherence of infected RBCs to endothelial receptors like ICAM-1 and CSA (these contribute to cerebral and placental malaria, respectively) in response to their CKII inhibition. Collectively, this study furthers our understanding of VARC function, underscores the importance of erythrocytic CKII in cytoadherence, and suggests a possible new target for anti-cytoadherence molecules. PMID- 19131329 TI - Dissection of the structural organization of the aminoacyl-tRNA synthetase complex. AB - The spatio-temporal organization of proteins within the cytoplasm of eukaryotic cells rests in part on the assembly of stable and transient multiprotein complexes. Here we examined the assembly of the multiaminoacyl-tRNA synthetase complex (MARS) in human cells. This complex contains nine aminoacyl-tRNA synthetases and three auxiliary proteins and is a hallmark of metazoan species. Isolation of the complexes has been performed by tandem affinity purification from human cells in culture. To understand the rules of assembly of this particle, expression of the three nonsynthetase components of MARS, p18, p38, and p43, was blocked by stable small interfering RNA silencing. The lack of these components was not lethal for the cells, but cell growth was slightly reduced. The residual complexes that could form in vivo in the absence of the auxiliary proteins were isolated by tandem affinity purification. From the repertoire of the subcomplexes that could be isolated, a comprehensive map of protein-protein interactions mediating complex assembly is deduced. The data are consistent with a structural role of the three nonsynthetase components of MARS, with p38 connecting two subcomplexes that may form in the absence of p38. PMID- 19131330 TI - Distinct regions within the erlins are required for oligomerization and association with high molecular weight complexes. AB - The group of stomatin/prohibitin/flotillin/HflK/C (SPFH) domain-containing proteins comprise members of diverse subcellular localization and function. Association with detergent-resistant membranes (DRMs) and the propensity to form oligomers are two common properties of SPFH domain proteins and likely important for the function of these proteins. Our laboratory recently discovered two novel members of this protein group, which, based on their endoplasmic reticulum (ER) localization and association with DRMs, were named ER lipid raft-associated protein (erlin)-1 and -2. Here we characterized erlin oligomerization and identified domains within the erlins responsible for oligomerization and DRM association. Using co-immunoprecipitation and sucrose density gradient centrifugation approaches on endogenous and ectopically expressed erlin proteins, we found that they formed homo- and hetero-oligomers and were part of large multimeric complexes. These properties were independent of their DRM association. By analyzing truncation and point mutants of erlin-2 we discovered that interaction between erlin monomers (oligomerization) and association with high molecular weight complexes require distinct regions within the protein. Although oligomerization and DRM association were mediated by a region immediately downstream of the SPFH domain (residues 228-300), integration into high molecular weight complexes was absolutely dependent on a phenylalanine residue C-terminal of this region (Phe-305), which lies within a short stretch of hydrophobic residues. Our data demonstrate that lower order oligomerization and incorporation into multimeric complexes are two separate biochemical properties of the erlins, because they are mediated by distinct regions. PMID- 19131331 TI - Protein kinase A activity controls the regulation of T-type CaV3.2 channels by Gbetagamma dimers. AB - Low voltage-activated (LVA), T-type, calcium channels mediate diverse biological functions and are inhibited by Gbetagamma dimers, yet the molecular events required for channel inhibition remain unknown. Here, we identify protein kinase A (PKA) as a molecular switch that allows Gbeta(2)gammax dimers to effect voltage independent inhibition of Ca(v)3.2 channels. Inhibition requires phosphorylation of Ser(1107), a critical serine residue on the II-III loop of the channel pore protein. S1107A prevents inhibition of unitary currents by recombinant Gbeta(2)gamma(2) dimers but does not disrupt dimer binding nor change its specificity. Gbetagamma dimers released upon receptor activation also require PKA activity for their inhibitory actions. Hence, dopamine inhibition of Ca(v)3.2 whole cell current is precluded by Gbetagamma-scavenger proteins or a peptide that blocks PKA catalytic activity. Fittingly, when used alone at receptor selective concentrations, D(1) or D(2) agonists do not elicit channel inhibition yet together synergize to inhibit Ca(v)3.2 channel currents. We propose that a dual-receptor regulatory mechanism is used by dopamine to control Ca(v)3.2 channel activity. This mechanism, for example, would be important in aldosterone producing adrenal glomerulosa cells where channel dysregulation would lead to overproduction of aldosterone and consequent cardiac, renal, and brain target organ damage. PMID- 19131332 TI - Use of a synthetic salicylic acid analog to investigate the roles of methyl salicylate and its esterases in plant disease resistance. AB - We previously demonstrated that salicylic acid-binding protein 2 (SABP2) of tobacco is an integral component of systemic acquired resistance (SAR). SABP2 is a methyl salicylate (MeSA) esterase that has high affinity for SA, which feedback inhibits its esterase activity. MeSA esterase activity is required in distal, healthy tissue of pathogen-infected plants to hydrolyze MeSA, which functions as a long-distance, phloem-mobile SAR signal; this hydrolysis releases the biologically active defense hormone SA. In this study, we examined the inhibitory interaction of SA with SABP2, and identified a synthetic SA analog, 2,2,2,2' tetra-f luoroacetophenone (tetraFA) that, like SA, competitively inhibits the activity of SABP2 and targets esterases, which utilize MeSA as a substrate. However, in contrast to SA, tetraFA does not induce downstream defense responses and, therefore, is effective in planta at blocking SAR development in tobacco mosaic virus (TMV)-infected tobacco and Pseudomonas syringae-infected Arabidopsis. These results confirm the importance of SABP2 and MeSA for SAR development in tobacco and establish similar roles for MeSA and the orthologs of SABP2 in Arabidopsis. Moreover, they demonstrate that tetraFA can be used to determine whether MeSA and its corresponding esterase(s) play a role in SAR signaling in other plant species. In planta analyses using tetraFA, in conjunction with leaf detachment assays and MeSA quantification, were used to assess the kinetics with which MeSA is generated in pathogen-infected leaves, transmitted through the phloem, and processed in the distal healthy leaves. In TMV-infected tobacco, these studies revealed that critical amounts of MeSA are generated, transmitted, and processed between 48 and 72 h post primary infection. PMID- 19131333 TI - In vitro interactions between the PII proteins and the nitrogenase regulatory enzymes dinitrogenase reductase ADP-ribosyltransferase (DraT) and dinitrogenase reductase-activating glycohydrolase (DraG) in Azospirillum brasilense. AB - The activity of the nitrogenase enzyme in the diazotroph Azospirillum brasilense is reversibly inactivated by ammonium through ADP-ribosylation of the nitrogenase NifH subunit. This process is catalyzed by DraT and is reversed by DraG, and the activities of both enzymes are regulated according to the levels of ammonium through direct interactions with the P(II) proteins GlnB and GlnZ. We have previously shown that DraG interacts with GlnZ both in vivo and in vitro and that DraT interacts with GlnB in vivo. We have now characterized the influence of P(II) uridylylation status and the P(II) effectors (ATP, ADP, and 2-oxoglutarate) on the in vitro formation of DraT-GlnB and DraG-GlnZ complexes. We observed that both interactions are maximized when P(II) proteins are de-uridylylated and when ADP is present. The DraT-GlnB complex formed in vivo was purified to homogeneity in the presence of ADP. The stoichiometry of the DraT-GlnB complex was determined by three independent approaches, all of which indicated a 1:1 stoichiometry (DraT monomer:GlnB trimer). Our results suggest that the intracellular fluctuation of the P(II) ligands ATP, ADP, and 2-oxoglutarate play a key role in the post translational regulation of nitrogenase activity. PMID- 19131334 TI - Yas3p, an Opi1 family transcription factor, regulates cytochrome P450 expression in response to n-alkanes in Yarrowia lipolytica. AB - In the alkane-assimilating yeast Yarrowia lipolytica, the expression of ALK1, a gene encoding cytochrome P450 that catalyzes the first step of n-alkane oxidation, is induced by n-alkanes. We previously demonstrated that two basic helix-loop-helix proteins, Yas1p and Yas2p, activate the transcription of ALK1 in an alkane-dependent manner by forming a heterocomplex and binding to alkane responsive element 1 (ARE1), a cis-acting element in the ALK1 promoter. Here we identified an Opi1 family transcription factor, Yas3p, involved in the alkane dependent transcription regulation of ALK genes. Deletion of YAS3 caused a significant increase in ALK1 mRNA in cells grown on glucose, glycerol, and n alkanes. The YAS3 deletion also resulted in a marked elevation of reporter gene expression driven by an ARE1-containing promoter on glycerol and n-decane. Bacterially expressed Yas3p bound specifically to Yas2p, but not to Yas1p, in vitro. In addition, although green fluorescent protein-tagged Yas3p was localized in the nucleus in glucose-containing medium, it changed its localization to an endoplasmic reticulum-like compartment upon transfer to medium containing n decane. These findings suggest that Yas3p functions as a master regulator of transcriptional response, which changes its localization between the nucleus and endoplasmic reticulum membrane in response to different carbon sources. Furthermore, quantitative real time PCR analysis of 12 ALK genes in YAS1, YAS2, and YAS3 deletion mutants suggested that Yas3p is involved in the transcriptional repression of a variety of ALK genes, including ALK1. In contrast, YAS3 deletion did not affect the mRNA level of an INO1 ortholog in Y. lipolytica, indicating functional diversity of Opi1 family transcription factors. PMID- 19131335 TI - Respiratory syncytial virus inhibits lung epithelial Na+ channels by up regulating inducible nitric-oxide synthase. AB - Respiratory syncytial virus (RSV) infection has been shown to reduce Na+-driven alveolar fluid clearance in BALB/c mice in vivo. To investigate the cellular mechanisms by which RSV inhibits amiloride-sensitive epithelial Na+ channels (ENaC), the main pathways through which Na+ ions enter lung epithelial cells, we infected human Clara-like lung (H441) cells with RSV that expresses green fluorescent protein (rRA2). 3-6 days later patch clamp recordings showed that infected cells (i.e. cells expressing green fluorescence; GFP+) had significantly lower whole-cell amiloride-sensitive currents and single channel activity (NPo) as compared with non-infected (GFP-), non-inoculated, or cells infected with UV inactivated RSV. Both alpha and beta ENaC mRNA levels were significantly reduced in GFP+ cells as measured by real-time reverse transcription-PCR. Infection with RSV increased expression of the inducible nitric-oxide synthase (iNOS) and nitrite concentration in the culture medium; nuclear translocation of NF-kappaB p65 subunit and NF-kappaB activation were also up-regulated. iNOS up-regulation in GFP+ cells was prevented by knocking down IkappaB kinase gamma before infection. Furthermore, pretreatment of H441 cells with the specific iNOS inhibitor 1400W (1 microM) resulted in a doubling of the amiloride-sensitive Na+ current in GFP+ cells. Additionally, preincubation of H441 cells with A77-1726 (20 microM), a de novo UTP synthesis inhibitor, and 1400W completely reversed the RSV inhibition of amiloride-sensitive currents in GFP+ cells. Thus, both UTP- and iNOS-generated reactive species contribute to ENaC down-regulation in RSV infected airway epithelial cells. PMID- 19131336 TI - An upstream open reading frame regulates translation of GADD34 during cellular stresses that induce eIF2alpha phosphorylation. AB - Cellular stress such as endoplasmic reticulum stress, hypoxia, and viral infection activates an integrated stress response, which includes the phosphorylation of the eukaryotic initiation factor 2alpha (eIF2alpha) to inhibit overall protein synthesis. Paradoxically, this leads to translation of a subset of mRNAs, like transcription factor ATF4, which in turn induces transcription of downstream stress-induced genes such as growth arrest DNA-inducible gene 34 (GADD34). GADD34 interacts with protein phosphatase 1 to dephosphorylate eIF2alpha, resulting in a negative feedback loop to recover protein synthesis and allow translation of stress-induced transcripts. Here, we show that GADD34 is not only transcriptionally induced but also translationally regulated to ensure maximal expression during eIF2alpha phosphorylation. GADD34 mRNAs are preferentially associated with polysomes during eIF2alpha phosphorylation, which is mediated by its 5'-untranslated region (5'UTR). The human GADD34 5'UTR contains two non-overlapping upstream open reading frames (uORFs), whereas the mouse version contains two overlapping and out of frame uORFs. Using 5'UTR GADD34 reporter constructs, we show that the downstream uORF mediates repression of basal translation and directs translation during eIF2alpha phosphorylation. Furthermore, we show that the upstream uORF is poorly translated and that a proportion of scanning ribosomes bypasses the upstream uORF to recognize the downstream uORF. These findings suggest that GADD34 translation is regulated by a unique 5'UTR uORF mechanism to ensure proper GADD34 expression during eIF2alpha phosphorylation. This mechanism may serve as a model for understanding how other 5'UTR uORF-containing mRNAs are regulated during cellular stress. PMID- 19131337 TI - Rational design of alpha-conotoxin analogues targeting alpha7 nicotinic acetylcholine receptors: improved antagonistic activity by incorporation of proline derivatives. AB - Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that belong to the superfamily of Cys loop receptors. Valuable insight into the orthosteric ligand binding to nAChRs in recent years has been obtained from the crystal structures of acetylcholine-binding proteins (AChBPs) that share significant sequence homology with the amino-terminal domains of the nAChRs. alpha-Conotoxins, which are isolated from the venom of carnivorous marine snails, selectively inhibit the signaling of neuronal nAChR subtypes. Co-crystal structures of alpha-conotoxins in complex with AChBP show that the side chain of a highly conserved proline residue in these toxins is oriented toward the hydrophobic binding pocket in the AChBP but does not have direct interactions with this pocket. In this study, we have designed and synthesized analogues of alpha-conotoxins ImI and PnIA[A10L], by introducing a range of substituents on the Pro(6) residue in these toxins to probe the importance of this residue for their binding to the nAChRs. Pharmacological characterization of the toxin analogues at the alpha(7) nAChR shows that although polar and charged groups on Pro(6) result in analogues with significantly reduced antagonistic activities, analogues with aromatic and hydrophobic substituents in the Pro(6) position exhibit moderate activity at the receptor. Interestingly, introduction of a 5-(R) phenyl substituent at Pro(6) in alpha-conotoxin ImI gives rise to a conotoxin analogue with a significantly higher binding affinity and antagonistic activity at the alpha(7) nAChR than those exhibited by the native conotoxin. PMID- 19131338 TI - Identification and characterization of a novel nuclear protein complex involved in nuclear hormone receptor-mediated gene regulation. AB - NRC/NCoA6 plays an important role in mediating the effects of ligand-bound nuclear hormone receptors as well as other transcription factors. NRC interacting factor 1 (NIF-1) was cloned as a novel factor that interacts in vivo with NRC. Although NIF-1 does not directly interact with nuclear hormone receptors, it enhances activation by nuclear hormone receptors presumably through its interaction with NRC. To further understand the cellular and biological function of NIF-1, we identified NIF-1-associated proteins by in-solution proteolysis followed by mass spectrometry. The identified components revealed factors involved in histone methylation and cell cycle control and include Ash2L, RbBP5, WDR5, HCF-1, DBC-1, and EMSY. Although the NIF-1 complex contains Ash2L, RbBP5, and WDR5, suggesting that the complex might methylate histone H3-Lys-4, we found that the complex contains a H3 methyltransferase activity that modifies a residue other than H3-Lys-4. The identified components form at least two distinctly sized NIF-1 complexes. DBC-1 and EMSY were identified as integral components of an NIF 1 complex of approximately 1.5 MDa and were found to play an important role in the regulation of nuclear receptor-mediated transcription. Stimulation of the Sox9 and HoxA1 genes by retinoic acid receptor-alpha was found to require both DBC-1 and EMSY in addition to NIF-1 for maximal transcriptional activation. Interestingly, NRC was not identified as a component of the NIF-1 complex, suggesting that NIF-1 and NRC do not exist as stable in vitro purified complexes, although the separate NIF-1 and NRC complexes appear to functionally interact in the cell. PMID- 19131339 TI - Oxidative stress promotes transcriptional up-regulation of Fyn in BCR-ABL1 expressing cells. AB - Signaling initiated by the BCR-ABL1 kinase causes chronic myelogenous leukemia (CML). Recently, we reported that expression of Fyn, a Src kinase, is heightened in CML cells and patient specimens and confers in vitro and in vivo proliferative advantages. Fyn is regulated by redox, and because BCR-ABL1 raises intracellular oxidant levels, which have been implicated in CML progression, we explored the molecular regulation of Fyn. Here we identify the transcription factors that drive redox- and BCR-ABL1-dependent Fyn expression. Promoter deletion analysis in 293T, BaF3, BaF3-p210, and K562 cells identified the region essential for basal transcriptional activity. Mutation of Sp1 and Egr1 binding sites within the essential region diminished Fyn promoter activity and identified Egr1 as conferring redox sensitivity. Gel shift and chromatin immunoprecipitation assays confirmed the binding of Sp1 and Egr1 to the promoter fragments. Importantly, knockdown of Sp1 or Egr1 with small interference RNA or inhibition of Sp1 binding by mithramycin A repressed Fyn protein expression. Our work is the first to define transcription factors that are responsible for endogenous, oxidative stress-dependent and BCR-ABL1-dependent Fyn expression. PMID- 19131340 TI - Atrophin proteins interact with the Fat1 cadherin and regulate migration and orientation in vascular smooth muscle cells. AB - Fat1, an atypical cadherin induced robustly after arterial injury, has significant effects on mammalian vascular smooth muscle cell (VSMC) growth and migration. The related Drosophila protein Fat interacts genetically and physically with Atrophin, a protein essential for development and control of cell polarity. We hypothesized that interactions between Fat1 and mammalian Atrophin (Atr) proteins might contribute to Fat1 effects on VSMCs. Like Fat1, mammalian Atr expression increased after arterial injury and in VSMCs stimulated with growth and chemotactic factors including angiotensin II, basic fibroblast growth factor, and platelet-derived growth factor BB. Two distinct Atr2 transcripts, atr2L and atr2S, were identified by Northern analysis; in VSMCs, atr2S mRNA expression was more responsive to stimuli. By immunocytochemistry, Fat1 and Atrs colocalized at cell-cell junctions, in the perinuclear area, and in the nucleus. In coimmunoprecipitation studies, Fat1 interacted with both Atr1 and Atr2; these interactions required Fat1 amino acids 4300-4400 and an intact Atro-box in the Atrs. Knock-down of Atrs by small interfering RNA did not affect VSMC growth but had complex effects on migration, which was impaired by Atr1 knockdown, enhanced by Atr2L knockdown, and unchanged when both Atr2S and Atr2L were depleted. Enhanced migration caused by Atr2L knockdown required Fat1 expression. Similarly, orientation of cells after monolayer denudation was impaired in cells with Atr1 knockdown but enhanced in cells selectively depleted of Atr2L. Together these findings suggest that Fat1 and Atrs act in concert after vascular injury but show further that distinct Atr isoforms have disparate effects on VSMC directional migration. PMID- 19131341 TI - The PN2-3 domain of centrosomal P4.1-associated protein implements a novel mechanism for tubulin sequestration. AB - Microtubules are cytoskeletal components involved in multiple cell functions such as mitosis, motility, or intracellular traffic. In vivo, these polymers made of alphabeta-tubulin nucleate mostly from the centrosome to establish the interphasic microtubule network or, during mitosis, the mitotic spindle. Centrosomal P4.1-associated protein (CPAP; also named CENPJ) is a centrosomal protein involved in the assembly of centrioles and important for the centrosome function. This protein contains a microtubule-destabilizing region referred to as PN2-3. Here we decrypt the microtubule destabilization activity of PN2-3 at the molecular level and show that it results from the sequestration of tubulin by PN2 3 in a non-polymerizable 1:1 complex. We also map the tubulin/PN2-3 interaction both on the PN2-3 sequence and on the tubulin surface. NMR and CD data on free PN2-3 in solution show that this is an intrinsically unstructured protein that comprises a 23-amino acid residue alpha-helix. This helix is embedded in a 76 residue region that interacts strongly with tubulin. The interference of PN2-3 with well characterized tubulin properties, namely GTPase activity, nucleotide exchange, vinblastine-induced self-assembly, and stathmin family protein binding, highlights the beta subunit surface located at the intermolecular longitudinal interface when tubulin is embedded in a microtubule as a tubulin/PN2-3 interaction area. These findings characterize the PN2-3 fragment of CPAP as a protein with an unprecedented tubulin sequestering mechanism distinct from that of stathmin family proteins. PMID- 19131343 TI - Effect of baroreflex stimulation using phenylephrine injection on ST segment elevation and ventricular arrhythmia-inducibility in Brugada syndrome patients. AB - AIMS: Patients affected by Brugada syndrome (BrS) are at risk of sudden cardiac death specifically at rest, when vagal tone is high. The aim of our study was to assess whether a phenylephrine injection, which provokes a baroreflex stimulation, could induce modification of the ST segment elevation and ventricular arrhythmias. METHODS AND RESULTS: Baroreflex test was performed with the administration of phenylephrine (2 microg/kg) to four highly symptomatic patients in a setting fully equipped for cardiac resuscitation. Phenylephrine injection induced a deep vagal stimulation with a decrease in the mean heart rate from 75 +/- 7 to 50 +/- 8 bpm and an increase in the mean systolic blood pressure from 141 +/- 14 to 204 +/- 46 mmHg. ST segment elevation was not modified and no ventricular arrhythmias were induced during the test. CONCLUSION: Although phenylephrine injection induced a major alpha-adrenergic vasoconstriction followed by an arterial baroreflex, this test failed to provoke ventricular arrhythmias or modification of the ST segment elevation in BrS patients. PMID- 19131342 TI - Biochemical, functional, and pharmacological characterization of AT-56, an orally active and selective inhibitor of lipocalin-type prostaglandin D synthase. AB - We report here that 4-dibenzo[a,d]cyclohepten-5-ylidene-1-[4-(2H-tetrazol-5-yl) butyl]-piperidine (AT-56) is an orally active and selective inhibitor of lipocalin-type prostaglandin (PG) D synthase (L-PGDS). AT-56 inhibited human and mouse L-PGDSs in a concentration (3-250 microm)-dependent manner but did not affect the activities of hematopoietic PGD synthase (H-PGDS), cyclooxygenase-1 and -2, and microsomal PGE synthase-1. AT-56 inhibited the L-PGDS activity in a competitive manner against the substrate PGH(2) (K(m) = 14 microm) with a K(i) value of 75 microm but did not inhibit the binding of 13-cis-retinoic acid, a nonsubstrate lipophilic ligand, to L-PGDS. NMR titration analysis revealed that AT-56 occupied the catalytic pocket, but not the retinoid-binding pocket, of L PGDS. AT-56 inhibited the production of PGD(2) by L-PGDS-expressing human TE-671 cells after stimulation with Ca(2+) ionophore (5 microm A23187) with an IC(50) value of about 3 microm without affecting their production of PGE(2) and PGF(2alpha) but had no effect on the PGD(2) production by H-PGDS-expressing human megakaryocytes. Orally administered AT-56 (<30 mg/kg body weight) decreased the PGD(2) production to 40% in the brain of H-PGDS-deficient mice after a stab wound injury in a dose-dependent manner without affecting the production of PGE(2) and PGF(2alpha) and also suppressed the accumulation of eosinophils and monocytes in the bronco-alveolar lavage fluid from the antigen-induced lung inflammation model of human L-PGDS-transgenic mice. PMID- 19131344 TI - Broad complex tachycardia and atrioventricular dissociation in a patient with heart transplant. PMID- 19131345 TI - WPW syndrome: the 'Rosetta stone' of rhythmology. The history of the Rosetta stone. AB - Prior to the 'discovery' of the Wolff-Parkinson-White syndrome, interpreting the respective phenomena was akin to reading hieroglyphic characters; thus a clear pathophysiological understanding and practical clinical diagnosis were impossible. The epochal work by Wolff, Parkinson, and White, which resulted in the electrophysiologically correct interpretation of circus movements as the cause of tachycardic rhythm disorders, can therefore indeed be compared to the deciphering of hieroglyphic writing by Champollion in 1822 with the aid of the Rosetta stone. After intensive archaeological and graphological examinations by the Society of Antiquaries, the Rosetta stone finally made its way to the British Museum, where it can still be viewed and admired today. PMID- 19131346 TI - Estimation of glomerular filtration rate: does haemoglobin discriminate between ageing and true CKD? AB - AIM: The aim of this study was to analyse the association between chronic kidney disease (CKD) defined by an estimated glomerular filtration rate (eGFR) <60 ml/min/ 1.73 m(2) and anaemia in older people. BACKGROUND: Guidelines focus on early identification and management of CKD to prevent CKD progression and cardiovascular disease. However, the significance of CKD classification using eGFR in older people is unclear. METHODS: Serum creatinine and haemoglobin from individuals attending non-nephrology outpatient clinics were extracted from the state pathology provider over a 4-month period. The associations between eGFR, gender, age and haemoglobin were explored. RESULTS: Serum creatinine in 9853 individual patients aged > or =15 years was available for analysis. Haemoglobin was simultaneously available in 8752 (88.8%) subjects. There was a negative relationship between age and median eGFR, and the slope of the regression line was -0.68 ml/min/year for males and -0.74 ml/min/year for females. Over 35% of individuals > or =65 years were classified as having CKD stage > or =3. Odds ratios for haemoglobin <100 g/l for an eGFR <15, 15-29 and 30-59 versus reference GFR > or =60 ml/ min/1.73 m(2) in subjects 25-44 years were 34.2 (30.7-37.7), 23.4 (20.2-26.6) and 7.2 (5.3-9.1), respectively. In comparison, these were 8.9 (6.7-11.1), 5.6 (4.9-7.3) and 1.6 (1.1-2.1), respectively, in subjects > or =65 years. In subjects > or =65 years, odds ratios for haemoglobin <100 g/l for an eGFR 30-44 and 45-59 ml/min/1.73 m(2) versus reference GFR > or =60 ml/min/1.73 m(2) were 1.9 (1.3-2.5) and 1.2 (0.7-1.7), respectively. CONCLUSIONS: An eGFR <60 ml/min/1.73 m(2) is very common in older people. Only an eGFR <45 ml/min/1.73 m(2) identified a smaller sub-group of older people with an increased prevalence of significant anaemia suggesting a clinically relevant disease. The benefits of identifying older people with an eGFR > or =45 ml/min/1.73 m(2) need to be determined. PMID- 19131347 TI - TRPV5 gene polymorphisms in renal hypercalciuria. AB - BACKGROUND: Kidney stone formation is a major socioeconomic problem in humans, involving pain, recurrent treatment and renal insufficiency. As most renal precipitates contain calcium as a major component, hypercalciuria is the main risk factor for renal stone formation. Different forms of hypercalciuria can be classified, which primarily arise from defects in the main organs involved in calcium homeostasis. A distinction can be made between renal, absorptive and resorptive hypercalciuria, originating from disturbed calcium handling in kidney, intestine and bone, respectively. A positive family history predisposes individuals to an increased risk of stone formation, which strongly indicates the involvement of genetic susceptibility factors. TRPV5 is the renal epithelial calcium channel that is the gatekeeper protein in active calcium reabsorption in the kidney. TRPV5 gene ablation in mice leads to severe hypercalciuria, implying that TRPV5 is an interesting candidate gene for renal hypercalciuria in humans. This study aims to identify and functionally characterize TRPV5 gene aberrations in patients with renal hypercalciuria. METHODS: The TRPV5 coding region and intron-exon boundaries were screened for gene mutations in 20 subjects displaying renal hypercalciuria after which identified non-synonymous polymorphisms were functionally characterized by patch-clamp analysis. Wild-type and TRPV5 channels including polymorphisms were transiently expressed in human embryonic kidney (HEK) 293 cells and functionally characterized by path-clamp analysis. RESULTS: Genotyping TRPV5 in renal hypercalciuria patients revealed three non-synonymous and five synonymous polymorphisms. Electrophysiological characterization of the TRPV5 mutants did not reveal significant functional changes compared to wild-type TRPV5 channel recordings. CONCLUSIONS: In this specific patient cohort, our data do not support a primary role for TRPV5 in the pathogenesis of renal hypercalciuria. However, TRPV5 cannot be excluded as a candidate gene in hypercalciuria. PMID- 19131348 TI - About the effect of low-molecular-weight heparin on platelet function in haemodialysis patients. PMID- 19131349 TI - Corin, atrial natriuretic peptide and hypertension. PMID- 19131350 TI - Rat mesangial cells exhibit sex-specific profibrotic and proinflammatory phenotypes. AB - BACKGROUND: Chronic renal disease progresses more rapidly in males compared to females. This study investigated whether there were any inherent differences between male and female mesangial cells that could contribute to this phenomenon and whether these differences could be modulated by sex hormones. METHODS: Experiments were carried out on cultured mesangial cells derived from adult male and female Wistar rat kidneys. Fibronectin, TNFalpha and IL-1beta levels were measured in control and macrophage-conditioned medium (MCM)-injured cells in the presence and absence of 17beta estradiol or testosterone. RESULTS: Male mesangial cells expressed higher baseline fibronectin levels compared to female cells. Similarly, basal levels of the proinflammatory cytokines TNFalpha and IL-1beta were higher in male cells. Fibronectin and IL-1beta levels were enhanced proportionately between the sexes in response to MCM stimulation, whilst the increase in TNFalpha levels was greater in MCM-stimulated female cells. Treatment with 10(-8) M estradiol down-regulated baseline fibronectin levels in female mesangial cells but had no effect on basal levels in male cells. Estradiol had no effect on MCM-stimulated fibronectin levels in female mesangial cells but further increased stimulated levels in male cells. Testosterone had no effect on basal fibronectin levels of either sex but further enhanced MCM-stimulated fibronectin levels in mesangial cells of both sexes. Sex hormone treatment had no effect on cytokine levels in male mesangial cells. However, in female cells estradiol decreased TNFalpha levels and increased IL-1beta levels, while testosterone increased the levels of both cytokines. CONCLUSION: These data would suggest that male mesangial cells inherently exhibit greater profibrotic and proinflammatory characteristics than female cells. The inherent gender phenotypes are further modulated by sex hormones. This sexual dimorphism in mesangial cells may play a contributory role in the faster rate of progression to end-stage renal disease in males. PMID- 19131351 TI - Diabetic foot syndrome and renal function in type 1 and 2 diabetes mellitus show close association. AB - BACKGROUND: Diabetic nephropathy and diabetic foot syndrome (DFS) are two major complications of diabetes. Surprisingly, little is known of a potential relationship between renal function and the development of DFS in patients with preterminal renal insufficiency. A retrospective cohort study at a single tertiary university centre caring for a large collective of patients with type 1 and 2 diabetes was performed. Patients and methods. All patients with type 1 or 2 diabetes from 1989 to 2007 on the electronic patient sheet who had standardized food examination, albuminuria and serum creatinine were analysed. A total number of 899 patients with type 1 and 4007 individuals with type 2 diabetes were studied. Estimated glomerular filtration rate (eGFR) was calculated according to the modified equation 7 MDRD formula. Patients were grouped into the chronic kidney disease (CKD) stages according to the eGFR and presence of albuminuria. DFS was classified according to Wagner as well as Armstrong stages. RESULTS: Forty-six patients (5.1%) of 899 patients with type 1 diabetes have active or a history of DFS. Patients with type 1 diabetes and DSF had significantly higher serum creatinine levels, lower eGFR, higher systolic blood pressure and higher HbA1c levels compared to those without DFS. There was a significant negative correlation between eGFR and the presence of DFS in patients with type 1 diabetes (r = -0.155, P < 0.01). In type 1 diabetes patients, there was a significant negative correlation (Spearman test) between eGFR and Wagner stages (r = -0.218, P = 0.01) as well as Armstrong stages (r = -0.255, P = 0.01). Multiple logistic regression analysis revealed a significant association between the presence of DFS and eGFR (odds ratio 0.696 per 10 ml/min increase, 95% confidence interval 0.627-0.773, P < 0.001). A total of 532 type 2 patients from 4007 patients had DFS (13.7%). Compared with type 2 patients without DFS, those with DFS were significantly older (P < 0.005), exhibited a higher HbA1c, had a longer duration of diabetes (P < 0.005), higher serum creatinine levels (P < 0.005) and a lower eGFR (P < 0.005). There was a significant negative correlation between the Wagner stages and eGFR (r = -0.104, P < 0.01) as well as Armstrong stages and eGFR (r = 0.125, P < 0.01) in all patients with type 2 diabetes (Spearman test). Multiple logistic regression analysis revealed a significant association between the presence of DFS and eGFR (odds ratio 0.873 per 10 ml/min increase, 95% confidence interval 0.842-0.904, P < 0.001). There were also significant associations between DFS and duration of diabetes as well as diastolic blood pressure. In addition, the Jonckheere-Terpstra test confirmed the decrease of eGFR with increasing Wagner and Armstrong stages in patients with type 2 diabetes. Smoking was not associated with a higher prevalence of DFS in type 1 and 2 diabetic patients. CONCLUSION: There was a strong association between the degree of renal function impairment and DFS in this observational study. Data show that diabetics with DFS undergo a higher incidence of amputation; thus, it should be recommended that diabetic patients with renal insufficiency should be regularly screened for the presence of DFS. PMID- 19131352 TI - Asymmetric dimethylarginine is closely associated with the development and progression of nephropathy in patients with type 2 diabetes. AB - BACKGROUND: Nitric oxide (NO) is thought to play an important role in the pathogenesis of diabetic nephropathy. We conducted a prospective, observational cohort study to explore the relationship between plasma levels of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, and the development and progression of nephropathy in patients with type 2 diabetes. METHODS: This was a hospital-based observational cohort study in Japanese type 2 diabetic patients with normoalbuminuria [urinary albumin-to-creatinine ratio (ACR) <30 mg/g creatinine] or microalbuminuria (30 < or = ACR <300 mg/g creatinine). The primary endpoint was the development or progression of diabetic nephropathy, based on transition from any given stage to a more advanced stage of albuminuria. RESULTS: We studied 225 diabetic patients, 81 women and 144 men, with a mean (+/-SD) age of 64 +/- 10 years. The majority (183) of patients were normoalbuminuric, with the remainder microalbuminuric (42). During the median follow-up period of 5.2 years, 27 normoalbuminuric and 10 microalbuminuric patients reached the primary endpoint. When patients were separated according to the median ADMA level (0.46 mumol/l), patients with higher ADMA levels had a greater incidence of reaching the endpoint (P = 0.014 by the log-rank test). In the multivariate Cox proportional hazard model, the hazard ratio for reaching the endpoint for patients with higher versus lower ADMA levels was 2.72 (95% confidence interval 1.25-5.95; P = 0.012). CONCLUSIONS: Higher plasma levels of ADMA may be a novel and potent predictor of the progression of nephropathy in adult Japanese type 2 diabetic patients. PMID- 19131353 TI - Haemodialysis patients' readiness to pursue live donor kidney transplantation. AB - BACKGROUND: Despite the benefits of live donor kidney transplantation (LDKT) over deceased donor kidney transplantation, patients hesitate to pursue this option. METHODS: A total of 324 transplant-eligible haemodialysis patients attending 14 dialysis facilities in Maryland, Northern Virginia and Southern Pennsylvania were asked about their stages of readiness to pursue LDKT, attitudes towards LKDT and demographics. Logistic regressions were used to test the effect of patients' attitudes and demographics on their stages of readiness to pursue LDKT. RESULTS: Fewer than half of the patients who had heard about LDKT were considering this option. Among patients considering LDKT, 26% had not talked to their loved ones about LDKT and 54% had not asked anyone for a kidney. Concerns about the surgical procedure for the donor were associated with a lower likelihood of considering LDKT (adjusted OR = 0.38; CI 0.18-0.79), talking about LDKT (adjusted OR = 0.38; CI 0.18-0.78) and asking for a kidney (adjusted OR = 0.14; CI 0.06-0.36). Being satisfied with the information the patient received about LDKT was associated with a higher likelihood of talking with someone about LDKT and asking for a kidney (adjusted OR = 2.26; CI 1.33-3.83 and 3.89; CI 1.78-8.51). Women and younger patients were more likely to talk with family/friends about LDKT (respectively, adjusted OR = 1.76; CI 1.26-2.47 and 0.97; CI 0.95-0.99) and to ask for a kidney (respectively, adjusted OR = 4.36; CI 2.57-7.40 and 0.97; CI 0.94-0.99). CONCLUSION: Tailored educational programmes considering the patient's stage of readiness and related attitudinal and demographic factors might help patients move towards LDKT. PMID- 19131354 TI - CD2AP mutations are associated with sporadic nephrotic syndrome and focal segmental glomerulosclerosis (FSGS). AB - BACKGROUND: CD2-associated protein (CD2AP) is a crucial protein for the slit diaphragm assembly and function. In spite of the fact that CD2AP knockout causes nephrotic syndrome in mice and the heterozygous +/- mouse is prone to proteinuria, little is known about the relevance of this molecule in human renal pathology. METHODS: A total of 80 Italian patients with idiopathic nephrotic syndrome were enrolled and screened for changes in the CD2AP gene. A normal control group of 200 healthy donors was also studied. The coding region of the CD2AP gene was analysed by polymerase chain reaction, denaturing high-performance liquid chromatography and sequencing. Peripheral blood mononuclear cells from patients with CD2AP mutations and from healthy donors were isolated by the Ficoll Hypaque gradient, and the CD2/CD2AP interaction was studied on T-lymphocytes by confocal laser scanning microscopy analysis. The expression levels of CD2AP, nephrin and podocin proteins were evaluated by indirect immunofluorescence on renal biopsies from a patient with p.delGlu525 mutation and from control subjects. Moreover, the effect of the p.K301M mutation on cell viability was evaluated by flow cytometry and annexin V/propidium iodide staining. RESULTS: Three heterozygous mutations (c.904A>T; c.1120A>G; c.1573delAGA) producing respectively aminoacidic changes (p.K301M, p.T374A) or a deletion in functional domains (p.delGlu525) were found in three unrelated patients. One (p.K301M) produced a lysine to methionine change in the third interactive SH3 domain (position 301) and resulted in the defective CD2-CD2AP interaction and clustering; the other (c.1573delAGA) caused the deletion of the glutamic acid in position 525 in the COOH-terminal region of binding with nephrin and was associated with down-modulation of CD2AP, podocin and nephrin glomerular expression. CONCLUSIONS: Our findings suggest that CD2AP mutations modify the interaction with CD2 in lymphocytes and alter the composition of the renal slit diaphragm. PMID- 19131355 TI - The mortality risk of overhydration in haemodialysis patients. AB - BACKGROUND: While cardiovascular events remain the primary form of mortality in haemodialysis (HD) patients, few centres are aware of the impact of the hydration status (HS). The aim of this study was to investigate how the magnitude of the prevailing overhydration influences long-term survival. METHODS: We measured the hydration status in 269 prevalent HD patients (28% diabetics, dialysis vintage = 41.2 +/- 70 months) in three European centres with a body composition monitor (BCM) that enables quantitative assessment of hydration status and body composition. The survival of these patients was ascertained after a follow-up period of 3.5 years. The cut off threshold for the definition of hyperhydration was set to 15% relative to the extracellular water (ECW), which represents an excess of ECW of approximately 2.5 l. Cox-proportional hazard models were used to compare survival according to the baseline hydration status for a set of demographic data, comorbid conditions and other predictors. RESULTS: The median hydration state (HS) before the HD treatment (DeltaHSpre) for all patients was 8.6 +/- 8.9%. The unadjusted gross annual mortality of all patients was 8.5%. The hyperhydrated subgroup (n = 58) presented DeltaHSpre = 19.9 +/- 5.3% and a gross mortality of 14.7%. The Cox adjusted hazard ratios (HRs) revealed that age (HRage = 1.05, 1/year; P < 0.001), systolic blood pressure (BPsys) (HRBPsys = 0.986 1/mmHg; P = 0.014), diabetes (HRDia = 2.766; P < 0.001), peripheral vascular disease (PVD) (HRPVD = 1.68; P = 0.045) and relative hydration status (DeltaHSpre) (HRDeltaHSpre = 2.102 P = 0.003) were the only significant predictors of mortality in our patient population. CONCLUSION: The results of our study indicate that the hydration state is an important and independent predictor of mortality in chronic HD patients secondary only to the presence of diabetes. We believe that it is essential to measure the hydration status objectively and quantitatively in order to obtain a more clearly defined assessment of the prognosis of haemodialysis patients. PMID- 19131356 TI - Phosphatidylglycerol depletion induces an increase in myxoxanthophyll biosynthetic activity in Synechocystis PCC6803 cells. AB - Phosphatidylglycerol (PG) depletion suppressed the oxygen-evolving activity of Synechocystis PCC6803 pgsA mutant cells. Shortage of PG led to decreased photosynthetic activity, which, similar to the effect of high light exposure, is likely to generate the production of reactive oxygen species (ROS) or free radicals. Protection of the PG-depleted cells against light-induced damage increased the echinenone and myxoxanthophyll content of the cells. The increased carotenoid content was localized in a soluble fraction of the cells as well as in isolated thylakoid and cytoplasmic membranes. The soluble carotenoid fraction contained carotene derivatives, which may bind to proteins. These carotene protein complexes are similar to orange carotenoid protein that is involved in yielding protection against free radicals and ROS. An increase in the content of myxoxanthophyll and echinenone upon PG depletion suggests that PG depletion regulates the biosynthetic pathway of specific carotenoids. PMID- 19131357 TI - Transcript profiling of an Arabidopsis PSEUDO RESPONSE REGULATOR arrhythmic triple mutant reveals a role for the circadian clock in cold stress response. AB - Arabidopsis PSEUDO RESPONSE REGULATOR (PRR) genes are components of the circadian clock mechanism. In order to understand the scope of genome-wide transcriptional regulation by PRR genes, a comparison survey of gene expression in wild-type Arabidopsis and a prr9-11 prr7-10 prr5-10 triple mutant (d975) using mRNA collected during late daytime was conducted using an Affymetrix ATH-1 GeneChip. The expression of 'night genes' increased and the expression of 'day genes' decreased toward the end of the diurnal light phase, but expression of these genes was essentially constant in d975. The expression levels of 'night genes' were lower, whereas the expression of 'day genes' was higher in d975 than in the wild type. Bioinformatics approaches have indicated that the set of up-regulated genes in d975 and the set of cold-responsive genes have significant overlap. We found that d975 is more tolerant to cold, high salinity and drought stresses than the wild type. In addition, dehydration-responsive element B1/C-repeat-binding factor (DREB1/CBF), which is expressed around mid-day, is more highly expressed in d975. Raffinose and L-proline accumulated at higher levels in d975 even when plants were grown under normal conditions. These results suggest that PRR9, PRR7 and PRR5 are involved in a mechanism that anticipates diurnal cold stress and which initiates a stress response by mediating cyclic expression of stress response genes, including DREB1/CBF. PMID- 19131358 TI - Proteomic analysis of salt-responsive proteins in the mangrove plant, Bruguiera gymnorhiza. AB - To identify key proteins in the regulation of salt tolerance in the mangrove plant Bruguiera gymnorhiza, proteome analysis of samples grown under conditions of salt stress was performed. Comparative two-dimensional electrophoresis revealed that two, three and one protein were differentially expressed in the main root, lateral root and leaf, respectively, in response to salt stress. Among these, three proteins were identified by internal peptide sequence analysis: fructose-1,6-bisphosphate (FBP) aldolase and a novel protein in the main root, and osmotin in the lateral root. These results suggest that FBP aldolase and osmotin play roles in salt tolerance mechanisms common to both glycophytes and mangrove plants. Osmotin was abundant at early time points following salt treatment and seems to play a role in initial osmotic adaptation in lateral roots of B. gymnorhiza under salt stress, but does not contribute towards adaptation to prolonged or continuous exposure to salt stress. The amounts of these proteins were not correlated with those of the respective mRNAs, as determined by microarray analysis. A novel salt-responsive protein, not previously detected by expressed sequence tag analysis or transcriptome analysis, was also identified in this proteomic approach, and may provide insight into the salt tolerance mechanism of the mangrove plant. This is the first report of proteome analysis with detailed analysis of main and lateral roots of mangrove plants under salt stress conditions. PMID- 19131359 TI - Alcohol use and cardiovascular health outcomes: a comparison across age and gender in the Winnipeg Health and Drinking Survey Cohort. AB - BACKGROUND: research has reliably demonstrated cardioprotection from regular alcohol use. Heavy episodic drinking (HED), however, negates these beneficial effects and increases the risk of cardiovascular disease (CVD). The impact of age on the health effects of episodic drinking has not been evaluated. OBJECTIVE: to examine the association between alcohol volume and pattern of consumption on the risk of cardiovascular morbidity and mortality across the lifespan. DESIGN AND SETTING: prospective, community-based cohort study of adults in Winnipeg, Manitoba, Canada. SUBJECTS: a total of 1,154 participants (580 men and 574 women) aged 18-64 surveyed at baseline (1990-91) on alcohol consumption levels and pattern of use. METHODS: usual alcohol consumption was measured using a quantity frequency approach. HED was estimated by asking participants how often they consumed eight or more drinks in one sitting in the past year. Questions were asked separately for wine, beer and spirits. Surveillance for cardiovascular events was conducted for 10 years (i.e. up to age 74 years). Diagnoses of CVD were obtained via health utilization records. Cox proportional hazard models were derived for both genders and for 'young adults' (baseline age 18-34), 'middle aged adults' (baseline age 35-49) and 'older adults' (baseline 50-64). Models were adjusted for marital status, cigarette smoking status and educational level. RESULTS: Reduced risk of CVD was associated with usual consumption, whereas an increased risk was associated with HED. Among male usual drinkers, cardioprotection was afforded only to middle and older age groups. The benefits of regular consumption were seen only in the youngest age group among women. The heaviest usual consumption category was associated with a decreased risk of CVD in men. Heavy episodic drinking increased the risk of coronary heart disease in middle-aged men and was marginally significant in middle-aged women. Risk of hypertension was elevated in older men with heavy episodic drinking. CONCLUSIONS: The well-established relationship between regular alcohol consumption and decreased risk of CVD may not become evident until middle age or older in men. Women may benefit from usual consumption at a much younger age. In both sexes, however, these beneficial effects of alcohol use are negated when alcohol is consumed in a heavy episodic drinking pattern, particularly for middle-aged and older men. PMID- 19131361 TI - Local cancellous bone grafting for osteonecrosis of the femoral head. AB - OBJECTIVE: This study reports a novel local bone graft technique for the treatment of osteonecrosis of femoral head (ONFH). MATERIALS AND METHODS: From 1998 to 2005, a procedure using local bone grafting was performed for the treatment of ONFH in 11 patients. The local bone grafts were obtained from the intertrochanteric region and then impacted into the necrotic lesion. A wire coil was inserted into the remaining space after the grafting. RESULTS: At a mean follow-up of 61 months (range, 30 to 103 months), all 5 ARCO stage IIC hips survived but 3 of the 6 ARCO stage IIIA hips failed. The overall clinical success rate was 73%. CONCLUSIONS: This study demonstrates that local cancellous bone grafting combined with a space-filling device implanted into the ONFH is a promising procedure and the wire coil showed no interference to the remodeling of the femoral head after grafting. PMID- 19131360 TI - Sub-physiological sarcoglycan expression contributes to compensatory muscle protection in mdx mice. AB - Sarcoglycans are a group of single-pass transmembrane glycoproteins. In striated muscle, sarcoglycans interact with dystrophin and other dystrophin-associated proteins (DAPs) to form the dystrophin-associated glycoprotein complex (DGC). The DGC protects the sarcolemma from contraction-induced injury. Duchenne muscular dystrophy (DMD) is caused by dystrophin gene mutations. In the absence of dystrophin, the DGC is disassembled from the sarcolemma. This initiates a chain reaction of muscle degeneration, necrosis, inflammation and fibrosis. In contrast to human patients, dystrophin-null mdx mice are only mildly affected. Enhanced muscle regeneration and the up-regulation of utrophin and integrin are thought to protect mdx muscle. Interestingly, trace amounts of sarcoglycans and other DAPs can be detected at the mdx sarcolemma. It is currently unclear whether sub physiological sarcoglycan expression also contributes to the mild phenotype in mdx mice. To answer this question, we generated delta-sarcoglycan/dystrophin double knockout mice (delta-Dko) in which residual sarcoglycans were completely eliminated from the sarcolemma. Interestingly, utrophin levels were further increased in these mice. However, enhanced utrophin expression did not mitigate disease. The clinical manifestation of delta-Dko mice was worse than that of mdx mice. They showed characteristic dystrophic signs, body emaciation and more macrophage infiltration. Their lifespan was reduced by 60%. Furthermore, delta Dko muscle generated significantly less absolute muscle force and became more susceptible to contraction-induced injury. Our results suggest that sub physiological sarcoglycan expression plays a critical role in ameliorating muscle disease in mdx mice. We speculate that low-level sarcoglycan expression may represent a useful strategy to palliate DMD. PMID- 19131362 TI - NPC1 repression contributes to lipid accumulation in human macrophages exposed to environmental aryl hydrocarbons. AB - AIMS: Aryl hydrocarbons (AHs), such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and benzo(a)pyrene (BP), are environmental contaminants promoting the development of atherosclerosis-related cardiovascular diseases. In order to identify molecular mechanisms involved in these effects, we have analysed AH-mediated regulation of the lipid trafficking Niemann-Pick type C1 protein (NPC1) and its contribution to AH-induced macrophage lipid accumulation. METHODS AND RESULTS: Exposure of primary human macrophages to TCDD and BP decreased NPC1 mRNA expression in a time-dependent manner. NPC1 protein expression and NPC1-related acid sphingomyelinase activity were reduced in parallel. NPC1 was also similarly down-regulated in mice exposed to BP. Moreover, TCDD and BP were demonstrated to trigger lipid accumulation in human macrophages, as assessed by Oil Red O and Nile Red staining and cholesterol determination. Such lipid loading occurred at least partly in endosomal/lysosomal compartments as demonstrated by immunolabelling of lipid vesicles by the lysosome-associated membrane protein 1. These cellular phenotypic effects were found to be similar to those triggered by knock-down of NPC1 expression using siRNAs and were counteracted by NPC1 overexpression, thus supporting the contribution of NPC1 to AH-mediated lipid accumulation in macrophages. Finally, both NPC1 down-expression and lipid accumulation in response to TCDD were found to be abolished through knock-down of the AH receptor (AHR), a ligand-activated transcription factor mediating many effects of AHs. CONCLUSION: Our data have shown that contaminants such as TCDD and BP repress NPC1 expression in macrophages in an AHR-dependent manner, which likely contributes to macrophage lipid accumulation caused by these environmental chemicals. Thus, NPC1 appears to be a new molecular target regulated by environmental AHs and putatively involved in their deleterious cardiovascular effects. PMID- 19131363 TI - Hypoxia inhibits vasoconstriction induced by metabotropic Ca2+ channel-induced Ca2+ release in mammalian coronary arteries. AB - AIMS: We have previously described in rat basilar arterial myocytes that in the absence of extracellular Ca(2+) influx, activation of L-type Ca(2+) channels stimulates a metabotropic cascade leading to Ca(2+) release from the sarcoplasmic reticulum (SR) and contraction [a calcium channel-induced Ca(2+) release (CCICR) mechanism]. On the other hand, it is known that hypoxia reduces Ca(2+) channel activity in coronary myocytes. In the present study, we have investigated whether CCICR is present in coronary arterial myocytes and whether arterial ring contraction induced by CCICR can be inhibited by hypoxia. METHODS AND RESULTS: Isometric force, arterial diameter, cytosolic [Ca(2+)] and electrical activity were recorded on mammalian (porcine, rat, and human) coronary artery preparations (dispersed myocytes, arterial rings, and intact arterial segments). In the absence of extracellular Ca(2+), Ca(2+) channel activation increased cytosolic [Ca(2+)] in isolated myocytes and contracted arterial rings. This contraction was suppressed by antagonists of L-type Ca(2+) channels and by inhibiting Ca(2+) release from the SR. Hypoxia induced dilatation of coronary arterial rings pre contracted by activation of Ca(2+) channels in the absence of extracellular Ca(2+). This effect was present although K(ATP) channels and Rho kinase were blocked by glibenclamide and Y27632, respectively. CONCLUSION: We show that Ca(2+) channel activation can induce metabotropic coronary arterial ring contraction in the absence of extracellular Ca(2+) and that this CCICR mechanism is inhibited by hypoxia. Thus, besides reduction of Ca(2+) entry through Ca(2+) channels, hypoxia seems to induce coronary vasorelaxation by inhibition of metabotropic CCICR. PMID- 19131364 TI - Angiotensin II downregulates the fatty acid oxidation pathway in adult rat cardiomyocytes via release of tumour necrosis factor-alpha. AB - AIMS: Advanced heart failure is often associated with reduced myocardial fatty acid oxidation capacity. We have previously observed that failing hearts of mice with overexpression of angiotensinogen in the myocardium exhibit marked reduction of key regulatory proteins of fatty acid oxidation. In the present study, we determined whether exposure of adult rat cardiac (ARC) myocytes to angiotensin II (Ang II) influences expression of fatty acid translocase, muscle-type carnitine palmitoyl transferase-I, and medium-chain acyl-CoA dehydrogenase. METHODS AND RESULTS: Ang II reduced mRNA expression of the three regulatory proteins in ARC myocytes during the entire 14-days culture period. However, protein expression and palmitate oxidation rate remained unaltered for 7 days, but subsequently markedly decreased. The decrease of protein expression and of fatty acid oxidation coincided with the onset of increased protein expression of tumour necrosis factor-alpha (TNF-alpha). The effect of Ang II was completely abolished by either blocking TNF-alpha formation through inhibition of reactive oxygen species-mediated activation of nuclear factor-kappaB or by neutralizing TNF-alpha with a specific antibody. Activation of peroxisome proliferator-activated receptor-alpha (PPARalpha) and PPARbeta/delta counteracted Ang II-mediated reduction of the fatty acid oxidation pathway. CONCLUSION: Prolonged exposure of cardiac myocytes to Ang II elicits downregulation of the fatty acid oxidation pathway mediated by enhanced synthesis of TNF-alpha. PMID- 19131365 TI - Chronic inhibition of phosphodiesterase 5 does not prevent pressure-overload induced right-ventricular remodelling. AB - AIMS: Inhibition of phosphodiesterase 5 (PDE5) decreases pulmonary pressure and improves symptoms in patients with pulmonary arterial hypertension. It is unclear however, whether inhibition of PDE5 can prevent myocardial remodelling during right-ventricular pressure overload. METHODS AND RESULTS: Right-ventricular pressure overload was produced in male rats in a pulmonary hypertension model (monocrotaline 60 mg/kg s.c.) or by surgical pulmonary artery banding. PDE5 inhibition using oral sildenafil (50 mg/kg/day in drinking water) or placebo was initiated 14 days after monocrotaline treatment and continued for 14 days until final examination. In the pulmonary artery banding groups, rats were treated with sildenafil (50 mg/kg/day) or placebo for 21 days following surgical pulmonary artery banding. At the final experiments, right-ventricular haemodynamics were measured and remodelling was analysed using histological, biochemical, and gene expression markers. Both monocrotaline and pulmonary artery banding increased right-ventricular systolic pressure to approximately 80 mmHg. In parallel, both interventions induced markers of hypertrophy (upregulation of natriuretic peptides, increase in myocyte diameter) and fibrosis (upregulation of collagen types 1A2 and 3A1) as well as mRNA expression of the tissue inhibitor of matrix metalloproteases 1 and osteopontin in the right ventricle. In monocrotaline model, sildenafil decreased pulmonary pressure, reduced right-ventricular hypertrophy, and prevented fibrosis marker gene upregulation. After pulmonary artery banding, in contrast, sildenafil increased markers of myocardial remodelling and right-ventricular myocyte diameter. CONCLUSION: Sildenafil prevents myocardial remodelling in pulmonary hypertension through an indirect action via right-ventricular unloading. PMID- 19131366 TI - Human microRNAs co-silence in well-separated groups and have different predicted essentialities. AB - BACKGROUND: Short regulating RNAs guide many cellular processes. Compared with transcription factor proteins they appear to provide more specialized control and their deletions are less frequently lethal. RESULTS: We find large differences between computationally predicted lists of human microRNA (miRNA)-target pairs. Instead of integrating these lists we use the two most accurate of them. Next, we construct the co-regulation network of human miRNAs as nodes by computing the correlation (link weight) between the gene silencing scores of individual miRNAs. In this network, we locate groups of tightly co-regulating nodes (modules). Despite explicitly allowing overlaps the co-regulation modules of miRNAs are well separated. We use the modules and miRNA co-expression data to define and compute miRNA essentiality. Instead of focusing on particular biological functions we identify a miRNA as essential, if it has a low co-expression with the miRNAs in its module. This may be thought of as having many workers performing the same tasks together in one place (non-essential miRNAs) as opposed to a single worker performing those tasks alone (essential miRNA). CONCLUSIONS: On the system level, we quantitatively confirm previous findings about the specialized control provided by miRNAs. For knock-out tests we list the groups of our predicted most and least essential miRNAs. In addition, we provide possible explanations for (i) the low number of individually essential miRNAs in Caenorhabdtits elegans and (ii) the high number of ubiquitous miRNAs influencing cell and tissue-specific miRNA expression patterns in mouse and human. PMID- 19131367 TI - Non-negative matrix factorization of gene expression profiles: a plug-in for BRB ArrayTools. AB - SUMMARY: Non-negative matrix factorization (NMF) is an increasingly used algorithm for the analysis of complex high-dimensional data. BRB-ArrayTools is a widely used software system for the analysis of gene expression data with almost 9000 registered users in over 65 countries. We have developed a NMF analysis plug in in BRB-ArrayTools for unsupervised sample clustering of microarray gene expression data. Our analysis tool also incorporates an algorithm for Semi-NMF which can handle both positive and negative elements for log-ratio data. Output includes a heat map of sample clusters and differentially expressed genes with extensive biological annotation. For comparison, output also includes the results of K-means clustering. AVAILABILITY: The NMF analysis plug-in is freely available in BRB-ArrayTools for non-commercial users. BRB-ArrayTools can be downloaded at http://linus.nci.nih.gov/BRB-ArrayTools.html. The algorithms used for NMF and Semi-NMF are available at ftp://linus.nci.nih.gov/pub/NMF. PMID- 19131368 TI - Refractory in-stent restenosis following carotid artery stenting: a case report and review of operative management. AB - In-stent restenosis following carotid artery stenting is a challenging problem that vascular surgeons will likely encounter with increasing frequency. The following describes a patient who developed progressive in-stent restenosis 3 years after carotid artery stenting, which was treated with operative therapy after failed balloon angioplasty. A review of the literature describing surgical approaches to the treatment of in-stent restenosis was also performed. PMID- 19131369 TI - Proinflammatory and anti-inflammatory cytokine balance in patients with abdominal aortic aneurysm and the impact of aneurysm size. AB - Abdominal aortic aneurysm is a common condition with high mortality due to rupture; however, the condition also is associated with nonaneurysmal cardiovascular mortality. A possible contributing mechanism for the cardiovascular mortality is an imbalance between the proinflammatory and anti inflammatory systemic response. In the present study, 78 patients with abdominal aortic aneurysm and 41 controls without aneurysm matched by age, gender and smoking habits were investigated. Cytokines such as interleukin-6, interleukin 10, and monocyte chemoattractant protein-1 were measured in plasma. There was significantly higher level of interleukin-6 in patients with AAA compared to controls. The interleukin-6/ interleukin-10 ratio was highest in patients with large compared to small abdominal aortic aneurysm. In conclusion, the present data indicate a proinflammatory response and a proinflammatory to antiinflammatory imbalance in patients with abdominal aortic aneurysm which is dependent by aneurysm size. PMID- 19131370 TI - The vascular and biochemical effects of cilostazol in diabetic patients with peripheral arterial disease. AB - OBJECTIVES: Cilostazol improves walking in patients with peripheral arterial disease (PAD). We hypothesized that cilostazol reduces diabetic complications in PAD patients. METHODS: Diabetic PAD patients were prospectively recruited to a randomized double-blinded, placebo-controlled trial, using cilostazol 100mg twice a day. Clinical assessment included ankle-brachial index, arterial compliance, peripheral transcutaneous oxygenation, treadmill walking distance and validated quality of life (QoL) questionnaires. Biochemical analyses included glucose and lipid profiles. All tests were at baseline, 6, and 24 weeks. RESULTS: 26 diabetic PAD patients (20 men) were recruited. Cilostazol improved absolute walking distance at 6 and 24 weeks (86.4% vs. 14.1%, P = .049; 143% vs. 23.2%, P = .086). Arterial compliance and lipid profiles improved as did some QoL indices for cilostazol at 6 and 24 weeks. Blood indices were similar at baseline and at follow-up points for both treatment groups. CONCLUSIONS: Cilostazol is a well tolerated and efficacious treatment, which improves claudication distances in diabetic PAD patients with further benefits in arterial compliance, lipid profiles, and QoL. PMID- 19131371 TI - Venous outflow obstruction with retroperitoneal Kaposi's sarcoma and treatment with inferior vena cava stenting. AB - A 26-year-old man presented with acute renal insufficiency, and severe lower extremity swelling. Computed tomographic scan revealed retroperitoneal lymphadenopathy encasing both ureters and the inferior vena cava. He underwent placement of ureteral stents to relieve the obstruction and afterward underwent lymph node biopsy, which revealed Kaposi's sarcoma. He subsequently was diagnosed with acquired immunodeficiency syndrome. Abdominal and lower extremity venous duplex ultrasound did not show any evidence of deep vein thrombosis. The inferior vena cava measured 3.5 mm in diameter and was encased by retroperitoneal lymphadenopathy. Bilateral transfemoral venography and intravascular ultrasound demonstrated significant compression of the inferior vena cava below the renal veins. Endovascular treatment was followed with primary stenting under intravascular ultrasound guidance. His symptoms improved with reduction in swelling. At 1-year follow-up, the patient was ambulatory with mild symptoms, and on venography the iliac vein and inferior vena cava stents were widely patent. PMID- 19131372 TI - Endovascular repair of innominate artery injury secondary to air rifle pellet: a case report and review of the literature. AB - OBJECTIVE: Decreased morbidity makes endovascular treatment preferable for certain central aortic and great vessel injuries. We present a case of penetrating innominate injury, describe considerations of a catheter-based approach, and provide follow-up of repair. METHODS: A case report and review of the literature. RESULTS: A 16-year-old man presented with an isolated innominate artery injury following an air rifle wound. Standard transfemoral approach was used to gain access the innominate artery. The injury was treated with an 8 x 35 mm, balloon-expandable, covered stent. Completion imaging confirmed a well positioned stent with exclusion of the injury and normal flow in distal vessels. There were no symptoms of stent migration or stenosis 1 year following the injury. CONCLUSIONS: Specific anatomic characteristics including its proximity to the carotid and vertebral arteries make the endovascular approach to the innominate artery unique. This case demonstrates the viability of catheter-based approaches in treating vascular injury. PMID- 19131373 TI - Endovascular treatment for traumatic popliteal artery pseudoaneurysms after knee arthroplasty. AB - Trauma to the popliteal artery is a recognized complication of knee arthroplasty (total knee arthroplasty). It can present in a variety of ways, one of which is the development of popliteal artery pseudoaneurysm. We report the successful endovascular management of 2 patients who developed popliteal artery pseudoaneurysms following total knee arthroplasty using covered stent grafts. From presented evidence, endovascular therapy is a safe treatment modality and it appears to be a viable alternative to open surgery. PMID- 19131374 TI - Autogenous arterial and venous reconstruction for femoral vein leiomyosarcoma--a case report. AB - This report describes the removal of a large, symptomatic leiyomyosarcoma arising from the proximal femoral vein necessitating removal of the femoral venous and arterial circulations. Reconstruction was accomplished with autologous vein and initial coverage with ipsilateral rectus abdominus flap. Persistent, early postoperative lymphatic leak and groin sepsis secondary to staph aureus was managed with reoperation and coverage with contralateral rectus abdominus muscle flap. Negative pressure dressing device was used as a wound management adjunct and splint thickness skin graft providing final successful coverage. Two years following the operation the patient was without evidence of disease, had a patent vascular reconstruction and a well healed groin. Femoral vein leiyomyosarcoma is a rare vascular tumor, which is especially challenging to manage in the proximal location. Successful outcome is predicated on revascularization with autologous vein and on a multidisciplinary approach using various soft tissue coverage strategies and wound management adjuncts. PMID- 19131375 TI - Has the implementation of EVAR for ruptured AAA improved outcomes? AB - OBJECTIVE: Previous studies have demonstrated improved outcomes with endovascular aneurysm repair (EVAR) for the treatment of ruptured abdominal aortic aneurysms (rAAA). However, these data may not be generalizable to all regions. METHODS: Retrospective cohort study (2001-2005) using state inpatient data. RESULTS: 5,176 patients underwent repair of AAA. 700 repairs were performed for rAAA (618 [88%] with open surgery (OS) and 82 [12%] with EVAR). Mortality for rAAA was similar for EVAR and OS (45.1% vs. 52.4%, P = 0.21). Lack of insurance (OR = 5.1; 95%CI: 1.7-15.2) was a predictor of mortality. Cost of repair for rAAA was greater for EVAR ($51,339 +/- 51,719 vs. $39,967 +/- 43,354, P = 0.03) and hospital LOS was similar (14.08 +/- 17.97 vs.13.42 +/- 18.18; P = 0.8). CONCLUSION: EVAR did not offer a survival benefit in the state, had a similar hospital LOS, and was significantly more expensive. Further evaluation exploring explanations for inferior outcomes by region are required as EVAR becomes more commonly implemented for rAAA. PMID- 19131376 TI - Using mixed methods to evaluate the Pediatric Lead Assessment Network Education Training Program (PLANET). AB - The Pediatric Lead Assessment Network Education Training Program (PLANET) is a peer-to-peer in-person 1-hr lead poisoning prevention educational program for health professionals. This evaluation was designed to determine the impact of the PLANET program. Evaluation methods included analyzing data from PLANET sign-in sheets, evaluation forms, pre/postknowledge tests, claims data, and focus groups (FGs) and interviews (IVs) with PLANET attendees and nonattendees. Claims data were used to compare blood lead testing rates for physicians attending and those not attending a PLANET program. Over 2,000 health professionals attended the 192 PLANET presentations delivered between June 2001 and December 2006; most were registered nurses or physicians. Written evaluations were overwhelmingly positive. Posttests indicated increased provider knowledge about childhood lead poisoning prevention, and assessment of blood lead testing rates showed higher testing rates for PLANET attendees. FG and IV participants suggesting improvements including using alternative delivery modes. PMID- 19131377 TI - Outcomes of multimodal training for healthcare professionals at an AIDS education and training center. AB - There is an ongoing need for continuing professional education (CPE) in the rapidly changing field of HIV care, but the best instructional methods remain a subject of debate. This study assessed the effects of training at an AIDS Education and Training Center (AETC) over an 18-month period. Health care professionals (HCP) who attended more than one training event showed small but significant improvements over time in HIV-related clinical practice behaviors. The type of training also predicted self-reported practice behavior, with interactive trainings and individual consultations associated with greater change, and intensive clinical training activities associated with a faster rate of change but not better scores on the self-reported behavior measure. Participants also reported high levels of satisfaction, knowledge improvement, and intention to change after each training event; however, these results were unrelated to whether trainees actually reported improved practice behavior. PMID- 19131378 TI - A generalized pollination system in the tropics: bats, birds and Aphelandra acanthus. AB - BACKGROUND AND AIMS: A number of different types of flower-visiting animals coexist in any given habitat. What evolutionary and ecological factors influence the subset of these that a given plant relies on for its pollination? Addressing this question requires a mechanistic understanding of the importance of different potential pollinators in terms of visitation rate (pollinator 'quantity') and effectiveness at transferring pollen (pollinator 'quality') is required. While bat-pollinated plants typically are highly specialized to bats, there are some instances of bat-pollinated plants that use other pollinators as well. These generalized exceptions tend to occur in habitats where bat 'quantity' is poor due to low or fluctuating bat densities. METHODS: Aphelandra acanthus occurs in tropical cloud forests with relatively high densities of bat visitors, yet displays a mix of floral syndrome characteristics, suggesting adaptation to multiple types of pollinators. To understand its pollination system better, aspects of its floral phenology and the 'quantity' and 'quality' components of pollination by its floral visitors are studied here. KEY RESULTS: Flowers were found to open and senesce throughout the day and night, although anther dehiscence was restricted to the late afternoon and night. Videotaping reveals that flowers are visited nocturnally by bats and moths, and diurnally by hummingbirds. Analysis of pollen deposition shows that bats regularly transfer large amounts of conspecific pollen, while hummingbirds occasionally transfer some pollen, and moths rarely do so. CONCLUSIONS: Hummingbirds and bats were comparable in terms of pollination 'quantity', while bats were the most effective in terms of 'quality'. Considering these components together, bats are responsible for approx. 70 % of A. acanthus pollination. However, bats also transferred remarkably large amounts of foreign pollen along with the conspecific grains (three of four grains were foreign). It is suggested that the negative effects of interspecific pollen transfer may decrease bat 'quality' for A. acanthus, and thus select for generalization on multiple pollinators instead of specialization on bats. PMID- 19131379 TI - Characterisation of three high-energy photon and fast neutron reference radiation fields. AB - Three reference radiation fields for the purpose of radiation protection were characterised: (1) radiation field R-F, consisting of photons in the energy range of about 6 and 7 MeV and a small neutron contamination; (2) radiation field R-C, consisting of photons with energies of about 4.4 MeV and neutrons with energies up to 2.65 MeV; (3) radiation field R-CF, consisting of photons in the energy range of about 1 and 7 MeV and neutrons with energies about 1.5 MeV. The radiation fields R-F and R-C have previously been defined in the ISO standard 4037. Their neutron components, however, have never been described accurately in the past. The new radiation field R-CF is proposed for the first time. This radiation field can, e.g., be used to calibrate tissue-equivalent proportional counters instruments for measurements at flight altitudes. PMID- 19131380 TI - Predicting gene expression level from relative codon usage bias: an application to Escherichia coli genome. AB - We present an expression measure of a gene, devised to predict the level of gene expression from relative codon bias (RCB). There are a number of measures currently in use that quantify codon usage in genes. Based on the hypothesis that gene expressivity and codon composition is strongly correlated, RCB has been defined to provide an intuitively meaningful measure of an extent of the codon preference in a gene. We outline a simple approach to assess the strength of RCB (RCBS) in genes as a guide to their likely expression levels and illustrate this with an analysis of Escherichia coli (E. coli) genome. Our efforts to quantitatively predict gene expression levels in E. coli met with a high level of success. Surprisingly, we observe a strong correlation between RCBS and protein length indicating natural selection in favour of the shorter genes to be expressed at higher level. The agreement of our result with high protein abundances, microarray data and radioactive data demonstrates that the genomic expression profile available in our method can be applied in a meaningful way to the study of cell physiology and also for more detailed studies of particular genes of interest. PMID- 19131381 TI - Neonatal screening for life threatening congenital heart disease. PMID- 19131382 TI - Outcomes of conduct problems in adolescence: 40 year follow-up of national cohort. AB - OBJECTIVE: To describe long term outcomes associated with externalising behaviour in adolescence, defined in this study as conduct problems reported by a teacher, in a population based sample. DESIGN: Longitudinal study from age 13-53. SETTING: The Medical Research Council National Survey of Health and Development (the British 1946 birth cohort). PARTICIPANTS: 3652 survey members assessed by their teachers for symptoms of externalising behaviour at age 13 and 15. MAIN OUTCOME MEASURES: Mental disorder, alcohol abuse, relationship difficulties, highest level of education, social class, unemployment, and financial difficulties at ages 36-53. RESULTS: 348 adolescents were identified with severe externalising behaviour, 1051 with mild externalising behaviour, and 2253 with no externalising behaviour. All negative outcomes measured in adulthood were more common in those with severe or mild externalising behaviour in adolescence, as rated by teachers, compared with those with no externalising behaviour. Adolescents with severe externalising behaviour were more likely to leave school without any qualifications (65.2%; adjusted odds ratio 4.0, 95% confidence interval 2.9 to 5.5), as were those with mild externalising behaviour (52.2%; 2.3, 1.9 to 2.8), compared with those with no externalising behaviour (30.8%). On a composite measure of global adversity throughout adulthood that included mental health, family life and relationships, and educational and economic problems, those with severe externalising behaviour scored significantly higher (40.1% in top quarter), as did those with mild externalising behaviour (28.3%), compared with those with no externalising behaviour (17.0%). CONCLUSIONS: Adolescents who exhibit externalising behaviour experience multiple social and health impairments that adversely affect them, their families, and society throughout adult life. PMID- 19131383 TI - Impact of pulse oximetry screening on the detection of duct dependent congenital heart disease: a Swedish prospective screening study in 39,821 newborns. AB - OBJECTIVE: To evaluate the use of pulse oximetry to screen for early detection of life threatening congenital heart disease. DESIGN: Prospective screening study with a new generation pulse oximeter before discharge from well baby nurseries in West Gotaland. Cohort study comparing the detection rate of duct dependent circulation in West Gotaland with that in other regions not using pulse oximetry screening. Deaths at home with undetected duct dependent circulation were included. SETTING: All 5 maternity units in West Gotaland and the supraregional referral centre for neonatal cardiac surgery. PARTICIPANTS: 39,821 screened babies born between 1 July 2004 and 31 March 2007. Total duct dependent circulation cohorts: West Gotaland n=60, other referring regions n=100. MAIN OUTCOME MEASURES: Sensitivity, specificity, positive and negative predictive values, and likelihood ratio for pulse oximetry screening and for neonatal physical examination alone. RESULTS: In West Gotaland 29 babies in well baby nurseries had duct dependent circulation undetected before neonatal discharge examination. In 13 cases, pulse oximetry showed oxygen saturations or =10.63 mg and for 70% of veterans exposed to the nerve agent sarin--658 (29%) of exposed veterans had cumulative exposures > or =15.0 mg min m(-3). Ninety-three per cent of veterans exposed to sulphur mustard were classified to a semi-quantitative scale of dermal effect--3771 (37%) had a vesicle or necrosed area, and 69% of veterans exposed to sarin could be categorized by change in blood cholinesterase activity- 1033 (31%) had a depression in cholinesterase activity of > or =30%. CONCLUSIONS: The experimental archive at Porton Down has proved to be a rich source of data on tests conducted between 1941 and 1989. It has been possible to categorize most veterans according to date of test, chemical group, chemical, type of protection and, for certain chemicals, level of exposure and/or degree of acute toxicity. These categorizations have been used to assign veterans to exposure groups for epidemiological analysis. PMID- 19131405 TI - Diffusion-weighted echo-planar MR imaging of primary parotid gland tumors: is a prediction of different histologic subtypes possible? AB - BACKGROUND AND PURPOSE: Our aim was to determine the value of echo-planar diffusion-weighted MR imaging (epiDWI) in differentiating various types of primary parotid gland tumors. MATERIALS AND METHODS: One hundred forty-nine consecutive patients with suspected tumors of the parotid gland were examined with an epiDWI sequence by using a 1.5T unit. Image analysis was performed by 2 radiologists independently, and the intraclass correlation coefficient was computed. Histologic diagnosis was obtained in every patient. For comparison of apparent diffusion coefficients (ADCs), a paired 2-tailed Student t test with a Bonferroni correction was used. RESULTS: In 136 patients, a primary parotid gland tumor was confirmed by histology. Among the observers, a high correlation was calculated (0.98). ADC values of pleomorphic adenomas were significantly higher than those of all other entities, except for myoepithelial adenomas (P = .054). ADC values of Warthin tumors were different from those of myoepithelial adenomas, lipomas, and salivary duct carcinomas (P < .001, 0.013, and .037, respectively). Mucoepidermoid carcinomas, acinic cell carcinomas, and basal cell adenocarcinomas were not differentiable from Warthin tumors (P = .094, .396, and .604, respectively). CONCLUSION: epiDWI has the potential to differentiate pleomorphic adenoma and myoepithelial adenomas from all other examined entities. Due to an overlap not only within the group of benign and malignant lesions but also between groups, diagnoses should not be addressed on the basis of ADC values solely. Therefore, further studies combining DWI, morphologic criteria, and probably other MR imaging techniques seem warranted. PMID- 19131407 TI - Clinical and angiographic results of coiling of 196 very small (< or = 3 mm) intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Coiling of very small (< or = 3 mm) aneurysms is considered controversial because of technical difficulties and a higher rate of procedural aneurysm ruptures. In this study, we report clinical and angiographic results of coiling of aneurysms 3 mm or smaller in comparison with larger aneurysms in a large, single-center cohort of patients. MATERIALS AND METHODS: Between 1995 and July 2008, a total of 1295 aneurysms were selectively occluded with coils. Of 1295 aneurysms, 196 (15.1%) in 187 patients were very small. Of 196 aneurysms, 149 (76%) had ruptured and 47 (24%) had not ruptured. There were 51 males (27%) and 136 females (73%). Mean age was 54.7 years (age range, 11-78 years). RESULTS: Procedural morbidity rate was 2.1% and mortality rate, 1.1%. Procedural rupture occurred in 15 of 196 aneurysms (7.7%). In 13 of 15 procedural ruptures, this had no adverse effect on outcome. Early recurrent hemorrhage of the coiled aneurysm occurred in 2 patients (1.1%). Compared with larger aneurysms, in very small aneurysms more often a procedural rupture occurred (7.7% versus 3.6%; P = .018). Procedural morbidity rate was lower (3.2% versus 5.5%), but this was not significant (P = .26). Retreatment rate consisted predominantly of clipping soon after incomplete coiling and was lower than in larger aneurysms (5.1% versus 10.0%; P = .041). Other characteristics were not significantly different. CONCLUSIONS: Coiling of very small aneurysms was technically feasible, with good results. Although procedural aneurysm rupture was significantly more frequent in very small aneurysms, this did not lead to increased overall morbidity and mortality rates. Retreatment rate was lower than for larger aneurysms. PMID- 19131406 TI - Susceptibility-weighted imaging: technical aspects and clinical applications, part 2. AB - SUMMARY: Susceptibility-weighted imaging (SWI) has continued to develop into a powerful clinical tool to visualize venous structures and iron in the brain and to study diverse pathologic conditions. SWI offers a unique contrast, different from spin attenuation, T1, T2, and T2* (see Susceptibility-Weighted Imaging: Technical Aspects and Clinical Applications, Part 1). In this clinical review (Part 2), we present a variety of neurovascular and neurodegenerative disease applications for SWI, covering trauma, stroke, cerebral amyloid angiopathy, venous anomalies, multiple sclerosis, and tumors. We conclude that SWI often offers complementary information valuable in the diagnosis and potential treatment of patients with neurologic disorders. PMID- 19131408 TI - Differences of clinical manifestations according to the patterns of brain lesions in acute encephalopathy with reduced diffusion in the bilateral hemispheres. AB - BACKGROUND AND PURPOSE: The precise clinical characteristics of acute encephalopathy with bilateral reduced diffusion are not fully understood. We compared clinical, laboratory, and neuroimaging findings according to the patterns of brain lesions among children with reduced diffusion in the bilateral hemispheres. MATERIALS AND METHODS: Nine patients were analyzed. The patterns of brain lesions were divided into diffuse lesions and central-sparing lesions. Diffuse lesions were defined as reduced diffusion in the whole cortex and/or subcortical white matter. Central-sparing lesions were defined as the lack of reduced diffusion in the areas around the bilateral Sylvian fissures. Clinical, laboratory, and neuroimaging findings were compared between groups. RESULTS: Five patients showed diffuse lesions and 4 showed central-sparing lesions. Coma was significantly more common in patients with diffuse lesions, whereas a biphasic clinical course was more common in those with central-sparing lesions. Outcome was worse in patients with diffuse lesions. Maximal aspartate aminotransferase, alanine aminotransferase, and kinase levels were also significantly higher in patients with diffuse lesions. In 2 patients with diffuse lesions, diffusion weighted images during the acute phase revealed reduced diffusion in the bilateral frontal and occipital areas, followed by diffuse lesions. No patient with central-sparing lesions showed MR imaging abnormalities during the acute phase. CONCLUSIONS: Clinical manifestations in patients with diffuse lesions were severe, whereas those in patients with central-sparing lesions were relatively mild. PMID- 19131409 TI - Kyphoplasty and vertebroplasty produce the same degree of height restoration. AB - BACKGROUND AND PURPOSE: There are few comparative studies regarding morphologic changes after kyphoplasty and vertebroplasty. The purpose of this study was to compare restoration of vertebral body height and wedge angle and cement leakage with kyphoplasty and vertebroplasty in osteoporotic compression fractures. MATERIALS AND METHODS: Forty patients (57 vertebrae) were treated with kyphoplasty, and 66 patients (124 vertebrae) were treated with vertebroplasty. Cement leakage into the disk space and paravertebral soft tissues or veins was analyzed on immediate postoperative CT scans. The height and wedge angle were measured before and after treatment and analyzed with the Mann-Whitney U test and chi(2) test. RESULTS: Kyphoplasty and vertebroplasty both improved vertebral body height and the wedge angles (P < .05). However, these differences were not statistically significant when the 2 techniques were compared (P > .05). There were 18% of the kyphoplasty group and 49% of the vertebroplasty group that showed cement leakage into the paravertebral soft tissues or veins (P < .01). Cement leakage into the disk space occurred in 12% of the kyphoplasty group and in 25% of the vertebroplasty group (P < .01). However, no complications related to cement leakage were noted. CONCLUSIONS: Both kyphoplasty and vertebroplasty achieved the same degree of height restoration and improvement of the wedge angle. Kyphoplasty resulted in less cement leakage into the disk space and paravertebral soft tissues or veins than vertebroplasty. PMID- 19131410 TI - Comparison of 2D digital subtraction angiography and 3D rotational angiography in the evaluation of dome-to-neck ratio. AB - BACKGROUND AND PURPOSE: Dome-to-neck ratio of intracranial aneurysms is an important predictor of outcomes of endovascular coiling. 3D imaging techniques are increasingly used in evaluating the dome-to-neck ratio of aneurysms for intervention. The purpose of this study was to determine whether 3D rotational angiography (3DRA) can be used to determine accurately the dome-to-neck ratio of intracranial aneurysms when compared with conventional 2D digital subtraction angiography (2D DSA). MATERIALS AND METHODS: A retrospective analysis of 180 patients with 205 intracranial aneurysms who underwent both 2D DSA and 3DRA for evaluation of previously untreated aneurysms was conducted. Dome-to-neck ratios were compared between 2D DSA and 3DRA images. The mean difference in dome-to-neck ratios between 2D DSA and 3DRA was calculated. The proportions of "wide-neck" aneurysms seen on 2D DSA and 3DRA were compared by using 2 different definitions of "wide-neck," including <1.5 and <2.0. RESULTS: The average dome-to-neck ratio was 1.81 +/- 0.55 and 1.55 +/- 0.48 for 2D DSA and 3DRA, respectively (P < .0001). When we defined "wide-neck" as a dome-to-neck ratio <1.5, sixty-nine (33.7%) aneurysms were wide-neck on 2D DSA compared with 119 (58%) on 3DRA (P < .0001). When we defined "wide-neck" as dome-to-neck ratio <2.0, one hundred forty two (69.3%) aneurysms were wide-neck on 2D DSA compared with 173 (84.4%) on 3DRA (P = .0004). CONCLUSIONS: In this retrospective study, 3DRA measurements resulted in significantly lower dome-to-neck ratios and significantly larger proportions of aneurysms defined as "wide-neck" compared with 2D DSA. Scrutiny of 2D DSA may offer substantial benefit over 3D techniques when triaging patients to or from endovascular therapy. PMID- 19131412 TI - Intra-arterial stroke therapy: an assessment of demand and available work force. AB - Intra-arterial therapy is currently applicable to a small subset of patients with ischemic stroke, but it will likely have an expanding role as new devices are introduced. This review evaluates the demand for such therapy and the physician work force available to provide such therapy in the United States. The available literature was reviewed to assess how many patients might need intra-arterial therapy annually and how many skilled neurointerventionalists are available to provide intra-arterial therapy for acute stroke. The number of acute ischemic strokes in the United States that will be amenable to intra-arterial therapy can only be crudely estimated, but it is certainly less than 126,000 per year and will quite likely be no more than 20,000 cases per year. The future demand for intra-arterial reperfusion techniques may change, but the number of patients who require intra-arterial thrombolysis is currently quite low. The overall number of neurointerventionists is currently adequate, though there might be local shortages. PMID- 19131411 TI - Hemodynamic patterns of anterior communicating artery aneurysms: a possible association with rupture. AB - BACKGROUND AND PURPOSE: The anterior communicating artery (AcomA) is a predilect location of aneurysms which typically carry higher rupture risks than other locations in the anterior circulation. The purpose of this study was to characterize the different flow types present in AcomA aneurysms and to investigate possible associations with rupture. MATERIALS AND METHODS: Patient specific computational models of 26 AcomA aneurysms were constructed from 3D rotational angiography images. Bilateral images were acquired in 15 patients who had both A1 segments of the anterior cerebral arteries, and models of the whole anterior circulation were created by fusing the reconstructed left and right arterial trees. Computational fluid dynamics simulations were performed under pulsatile flow conditions measured on a healthy subject. Visualizations of flow velocity, instantaneous streamlines, and wall shear stress (WSS) were performed. These were analyzed for flow patterns, size of the impaction zone, and peak WSS and then correlations were made with prior history of rupture. RESULTS: Aneurysms with small impaction zones were more likely to have ruptured than those with large impaction zones (83% versus 63%). Maximum intra-aneurysmal WSS (MWSS) for the unruptured aneurysms ranged from 10 to 230 dyne/cm(2) (mean, 114 dyne/cm(2)) compared with ruptured aneurysms, which ranged from 35 to 1500 dyne/cm(2) (mean, 271 dyne/cm(2)). This difference in MWSS was statistically significant at 90% confidence levels (P = .10). CONCLUSIONS: Aneurysms with small impaction zones, higher flow rates entering the aneurysm, and elevated MWSS are associated with a clinical history of previous rupture. PMID- 19131413 TI - Pediatric MR imaging with automated spine survey iterative scan technique (ASSIST). AB - MR imaging automated spine survey iterative scan technique (ASSIST) provides an automated subminute, submillimeter, in-plane resolution survey of the entire spine in 2 contiguous sagittal fast gradient-echo breath-hold series with computer labeling of vertebrae and disks. The technique was prospectively tested in 13 school-aged children for a wide range of clinical indications. In all cases, imaging was successful without requiring repeated sequencing. In all but 1 patient manifesting prominent scoliosis, automated labeling was concordant with neuroradiologist assignments. PMID- 19131414 TI - Abnormal white matter signal on MR imaging is related to abnormal tissue microstructure. AB - BACKGROUND AND PURPOSE: White matter signal-intensity abnormalities (WMSA) on MR imaging are related to adverse neurodevelopmental outcome in extremely preterm infants. Diffusion tensor imaging (DTI) may detect alterations in cerebral white matter microstructure and thus may help confirm the pathologic basis of WMSA. This study aimed to relate regional DTI measures with severity of WMSA in very preterm infants. MATERIALS AND METHODS: One hundred eleven preterm infants (birth weight, <1250 g and/or gestational age, <30 weeks) were scanned at term equivalent age (1.5T). WMSA were classified as normal, focal, or extensive. Apparent diffusion coefficient (ADC), fractional anisotropy (FA), axial (lambda1), and radial ([lambda2 + lambda3]/2) diffusivity were calculated in 12 regions of interest placed in the bilateral posterior limbs of the internal capsule, frontal (superior and inferior), sensorimotor, and occipital (superior and inferior) white matter regions. Data were compared by using 1-way analysis of variance, with a Bonferroni correction for multiple comparisons. RESULTS: Thirty nine infants had normal, 59 infants had focal, and 13 infants had extensive WMSA. Compared with infants with normal or focal WMSA, infants with extensive WMSA had significantly lower FA in the internal capsule (P < .001), right inferior frontal regions (P < .05), and right superior occipital regions (P = .01); and higher radial diffusivity in the right internal capsule (P = .005), bilateral sensorimotor (P < .05), and right superior occipital regions (P < .05). Compared with infants with normal WMSA, infants with extensive WMSA had significantly higher ADC in bilateral sensorimotor regions (P < .01) and right superior occipital regions (P = .01), and lower axial diffusivity in the bilateral sensorimotor regions (P < .05). CONCLUSIONS: There are significant region specific changes in ADC, FA, radial diffusivity, and axial diffusivity in preterm infants with extensive WMSA. Altered radial diffusivity was most prominent. This implies that disrupted premyelinating oligodendroglia is the major correlate with extensive WMSA rather than axonal pathology. PMID- 19131415 TI - Enlargement and transformation of thyroglossal duct cysts in response to radiotherapy: imaging findings. AB - BACKGROUND AND PURPOSE: Thyroglossal duct cyst remnants (TGDC) are a common midline neck mass that is mostly encountered in childhood. Enlargement after an upper respiratory tract infection or, rarely, cancer within TGDC, is a well documented phenomenon. In the pediatric population, it rarely presents a diagnostic quandary. However, in an adult population, especially one treated for head and neck cancer with radiation therapy (RT), any enlarging cystic mass would be concerning and would raise the specter of metastatic disease. We propose that inflammation associated with RT results in secretory stimulation and/or obstruction of a persistent thyroglossal duct, resulting in changes within a pre existing TGDC. MATERIALS AND METHODS: We present a case series of 8 patients with subclinical TGDC treated with RT for various head and neck cancers, in which we radiographically observed enlargement or cystic transformation (more cystlike appearance) of the cyst. RESULTS: In all patients there was in retrospect a small thyroglossal remnant, though not necessarily a discernible cyst, that was observed after radiotherapy to enlarge and become more cyst-like. CONCLUSIONS: In patients with pre-existing TGDC undergoing RT to the neck, enlargement and cystic transformation may be encountered radiographically and should not be misinterpreted as a tumor. Our series bolsters a recently reported description of the same phenomenon. PMID- 19131416 TI - Complications related to percutaneous transarterial embolization of intracranial dural arteriovenous fistulas in 40 patients. AB - BACKGROUND AND PURPOSE: An increasing number of intracranial dural arteriovenous fistulas (DAVFs) are amenable to endovascular treatment with Onyx. However, reports on complications caused by this technique have been limited. We present the initial Beijing Tiantan Hospital experience with adverse events related to transarterial Onyx embolization for DAVFs. MATERIALS AND METHODS: Between September 2005 and February 2008, a total of 40 patients with DAVFs were treated at our institute with Onyx-18. There were 11 women and 29 men with a mean age of 43.15 years (age range, 23-60 years). We reviewed the clinical presentation, angiographic features, treatment, and outcome. RESULTS: In 40 patients, total obliteration was achieved in 25 DAVFs (62.5%), with the remaining 15 patients not cured with residual shunts. Complications occurred in 9 patients, 5 DAVFs were located at tentorium, 2 were located at the transverse-sigmoid sinus, 1 was found at the inferior petrosal sinus, and 1 was found at the cavernous sinus, leading to permanent disability in 3 patients (morbidity, 7.5%). Complications included reflexive bradyarrhythmia in 3 (7.5%) patients, hemifacial hypoesthesia in 3 (7.5%) patients, hemifacial palsy in 2 (5%) patients, posterior fossa infarction in 2 (5%) patients, jaw pain in 1 (2.5%) patient, microcatheter gluing in 1 (2.5%) patient, hallucinations in 1 (2.5%) patient, and Onyx migration in 1 (2.5%) patient. CONCLUSION: Although complete cure can be achieved by transarterial embolization with Onyx, the potential for serious complications exists with this procedure. PMID- 19131417 TI - The influence of types of decision support on physicians' decision making. AB - OBJECTIVE: To determine whether physicians' post-test probability estimates are influenced by receiving test characteristics and impact their subsequent clinical decisions. DESIGN: Questionnaire based randomised controlled trial. SETTING: Mailed survey with a vignette describing an infant whose pretest likelihood of pertussis was 30% and direct fluorescent-antibody (DFA) test was negative for pertussis. SUBJECTS: Nationally representative sample of US paediatricians (n = 1502). INTERVENTIONS: Random receipt of no additional information (controls), the DFA's sensitivity and specificity (TC group) or the test's sensitivity and specificity with their definitions (TCD group). MAIN OUTCOME MEASURES: Estimated post-test probability (PTP) of pertussis, PTP of 0.50, "nearly correct" PTP (+/ 5%), intended erythromycin management and intended hospital disposition. ANALYSES: Chi2 and t tests. RESULTS: Despite the negative DFA result, 67% of the 635 (49.7%) participants who responded estimated a PTP higher than the pretest probability of 30%; the overall mean estimated PTP was 0.41 (SD 0.26) (correct answer: 0.18). The TCD group's mean PTP was significantly higher than controls' mean PTP (0.45 vs 0.38, p<0.001), while the TC and control groups' mean PTP did not differ significantly (0.41 vs 0.38, p = 0.16). With decision support significantly more TC and TCD participants compared to controls estimated the PTP as 0.50 (38% vs 17%, p<0.001; 41% vs 17%, p<0.001, respectively) and also estimated a nearly correct PTP more often (20% vs 13%, p = 0.06; 19% vs 13%, p = 0.08, respectively). The mean PTP of participants intending to discontinue erythromycin therapy or discharge the patient home was significantly lower than that of participants who intended continuing erythromycin or hospitalisation (0.20 vs 0.43, p<0.001; 0.40 vs 0.49, p = 0.005, respectively). CONCLUSIONS: Paediatricians differed in their response to information about test characteristics. For many, it increased errors in estimating post-test probability; for others, it reduced errors. Estimated post-test probability was logically associated with intended clinical management. PMID- 19131418 TI - Delivery of the paediatric curriculum of the Royal College of Paediatrics and Child Health (RCPCH). PMID- 19131419 TI - Emergency management of children with severe sepsis in the United Kingdom: the results of the Paediatric Intensive Care Society sepsis audit. AB - OBJECTIVE: To audit current UK practice of the management of severe sepsis in children against the 2002 American College of Critical Care Medicine/Pediatric Advanced Life Support (ACCM-PALS) guideline. DESIGN: Prospective observational study. SETTING: 17 UK paediatric intensive care units (PICUs) and two UK PICU transport services. PARTICIPANTS: 200 children accepted for PICU admission within 12 h of arrival in hospital, whether or not successfully transported to a PICU, with a discharge diagnosis of sepsis or suspected sepsis. MAIN OUTCOME MEASURES: Medical interventions, physiological and laboratory data to determine the presence or absence of shock, inter-hospital transfer times, predicted mortality (using the Paediatric Index of Mortality, version 2 (PIM2) scoring system) and observed mortality. RESULTS: 34/200 (17%) children died following referral. Although children defined as being in shock received significantly more fluid (p<0.001) than those who were not in shock, overall fluid and inotrope management suggested by the 2002 ACCM-PALS guideline was not followed in 62% of shocked children. Binary logistic regression analysis demonstrated that the odds ratio for death, if shock was present at PICU admission, was 3.8 (95% CI 1.4 to 10.2, p = 0.008). CONCLUSIONS: The presence of shock at PICU admission is associated with an increased risk of death. Despite clear consensus guidelines for the emergency management of children with severe sepsis and septic shock, most children received inadequate fluid resuscitation and inotropic support in the crucial few hours following presentation. PMID- 19131420 TI - Effect of adenoidectomy on respiratory function: a randomised prospective study. AB - OBJECTIVE: Risk of childhood asthma is increased in children with recurrent otitis media. This may be associated with recurrent respiratory tract infections in these children, but the role of adenoidectomy, a frequent operation during childhood, is unknown. Therefore, the role of adenoidectomy in the development of atopy and respiratory function changes characteristic of asthma was evaluated. DESIGN: Randomised controlled study. SETTING: Tertiary care centre. PATIENTS: 166 children aged 12-48 months who had recurrent or persistent otitis media and who were followed-up for 3 years after randomisation. INTERVENTION: Randomisation to undergo insertion of tympanostomy tubes with or without adenoidectomy. MAIN OUTCOME MEASURES: The primary outcome measure was exercise-induced bronchoconstriction as evaluated by impulse oscillometry. The secondary outcome measures were bronchial inflammation as evaluated by exhaled nitric oxide and atopy as evaluated by skin prick tests. During the 3-year follow-up period otitis media episodes were documented in patient diaries. RESULTS: Adenoidectomy did not significantly influence baseline lung function, exercise-induced bronchoconstriction, exhaled nitric oxide concentration, the development of positive skin prick tests, or doctor-diagnosed asthma. Adenoidectomy did not significantly prevent otitis media. Recurrent otitis media (>or=4 episodes) during the first follow-up year was associated with an abnormal exercise-induced bronchoconstriction (OR 6.62, 95% CI 1.27 to 34) and an elevated exhaled nitric oxide concentration (OR 3.26, 95% CI 0.98 to 10.8) regardless of adenoidectomy. CONCLUSIONS: Adenoidectomy did not promote asthma or allergy. Recurrent respiratory tract infections during early childhood are associated with the risk of bronchial hyper-reactivity. PMID- 19131422 TI - The incidence and aetiology of epistaxis in infants: a population-based study. AB - OBJECTIVE: To estimate the incidence and describe the aetiology of epistaxis in infants. DESIGN: Population-based study including a retrospective hospital admission database analysis and a postal questionnaire to clinicians. SETTING: Wales, United Kingdom. METHODS: Cases of epistaxis over a 6-year period (1999 2004) were identified from the Patient Episode Dataset for Wales (PEDW) and validated using clinical information to calculate the population-based incidence and ascertain the causes of epistaxis in infants in Wales. RESULTS: 36 confirmed cases were identified over the 6-year period giving an estimated annual incidence of epistaxis of 19.3 (95% CI 14.0 to 26.7) per 100,000 infants. The median age at admission was 12 weeks (interquartile range 4-33) (min 1 week, max 49 weeks). 23 of the infants had a recognised cause for their epistaxis (trauma (five), coagulation disorder (four), congenital anomaly (two), acute rhinitis or coryza (11), abusive smothering event (one)). No cause for the epistaxis was identified for 13 cases. Coagulation disorder was excluded in seven of these 13 infants but in the other six no attempt was made to exclude this disorder. Child abuse was suspected but excluded in four of the 13 cases. CONCLUSION: Hospital admission for epistaxis is a rare event. In the majority of cases in this study a simple explanation was available and proven physical abuse was rare. A bleeding disorder should always be considered and, if additional evidence suggests physical abuse, this must be excluded. PMID- 19131421 TI - UK childhood exposures to pesticides 2004-2007: a TOXBASE toxicovigilance study. AB - OBJECTIVE: There are no systematic methods for toxicovigilance of non-medicinal products in the UK. This is particularly relevant for pesticides, where there is significant public concern about potential adverse effects. This study describes a prospective toxicovigilance scheme based on follow-up of enquiries to the National Poisons Information Service (NPIS) through its online poisons information system TOXBASE. These enquiries reflect acute exposures and the patterns of acute illness that result. RESULTS: A total of 10 061 pesticide related enquiries were identified. After follow-up, data were gathered on 2364 suspected exposures, of which 1162 involved children. After exclusions, 1147 exposures are reported here. No deaths were reported and only 37 children were admitted to hospital. The majority were considered to have either minimal or no features (925, 80.6%). Symptoms for 38 children were unknown. Symptoms reported in the other 184 children included nausea or vomiting (58), eye irritation, pain or conjunctivitis (29), skin irritation (28), abdominal pain (24), mouth or throat irritation (18) and diarrhoea (15). Where age was recorded, 60.5% (680) of children involved in suspected pesticide exposures were aged 2 years or less. The most common scenario for acute accidental exposure to pesticide in children was exposure after application (329, 28.7%) or due to poor storage (228, 19.9%). CONCLUSIONS: Areas of potential concern identified included storage, access of young children to "laid" baits and pesticides, and exposures as a result of medication errors, with liquid head lice preparations being confused with other medicines. Use of NPIS systems provides a potentially useful method of toxicovigilance. PMID- 19131423 TI - Benign recurrent sixth (abducens) nerve palsies in children. AB - Sixth nerve palsy can occur as a result of elevated intracranial pressure, neoplasm or trauma. Reports from tertiary centres indicate that between 5% and 16% of referred cases have no ascribed aetiology and are classified as benign. Rarely, these benign palsies can recur. A retrospective chart review of a cohort of 253 paediatric patients with sixth nerve palsies was analysed and uncovered 30 cases of benign sixth nerve palsy, nine of which recurred. Our data and review of other studies on the subject imply that a new onset sixth nerve palsy presenting in children can be benign in approximately 13% of cases, so a thorough history and physical examination to evaluate for any other neurological symptoms or signs followed by MRI of the brain with and without contrast is recommended. PMID- 19131424 TI - Characterization of a novel macrolide efflux gene, mef(B), found linked to sul3 in porcine Escherichia coli. AB - OBJECTIVES: The aim of this study was to characterize a putative novel macrolide efflux gene located in the vicinity of sul3 in porcine Escherichia coli. METHODS: Five sul3-encoding E. coli isolates of porcine origin were investigated by plasmid characterization and random amplification of polymorphic DNA (RAPD) PCR. Unknown DNA adjacent to the sul3 genes was amplified using a PCR approach, followed by sequencing of the fragments. The putative macrolide efflux gene was cloned into pK18. The cloned gene was characterized by susceptibility testing by Etest in the presence and absence of efflux inhibitors. RESULTS: Five sul3 encoding isolates, demonstrated to be unrelated by RAPD PCR, were characterized. The immediate genetic context of sul3 in five isolates was identical to that in plasmid pVP440, and in all cases, sul3 was associated with class 1 integrons. In three isolates, an open reading frame (orf2) encoding a putative protein with 38% amino acid identity to Mef(A) was found, while the two remaining isolates contained a fragment of orf2 truncated by IS26 insertion. In three of the isolates, this DNA region was demonstrated to be located on non-conjugative plasmids. When the complete orf2 was cloned, it conferred high-level resistance to erythromycin and azithromycin, and the resistance property could be partially inhibited using the efflux inhibitor Phe-Arg beta-naphthylamide dihydrochloride. The gene was named mef(B). CONCLUSIONS: A new macrolide efflux protein, Mef(B), with 38% amino acid identity to Mef(A), has been characterized and represents the second member of the mef family of genes. PMID- 19131426 TI - Learning to count: robust estimates for labeled distances between molecular sequences. AB - Researchers routinely estimate distances between molecular sequences using continuous-time Markov chain models. We present a new method, robust counting, that protects against the possibly severe bias arising from model misspecification. We achieve this robustness by generalizing the conventional distance estimation to incorporate the empirical distribution of site patterns found in the observed pairwise sequence alignment. Our flexible framework allows for computing distances based only on a subset of possible substitutions. From this, we show how to estimate labeled codon distances, such as expected numbers of synonymous or nonsynonymous substitutions. We present two simulation studies. The first compares the relative bias and variance of conventional and robust labeled nucleotide estimators. In the second simulation, we demonstrate that robust counting furnishes accurate synonymous and nonsynonymous distance estimates based only on easy-to-fit models of nucleotide substitution, bypassing the need for computationally expensive codon models. We conclude with three empirical examples. In the first two examples, we investigate the evolutionary dynamics of the influenza A hemagglutinin gene using labeled codon distances. In the final example, we demonstrate the advantages of using robust synonymous distances to alleviate the effect of convergent evolution on phylogenetic analysis of an HIV transmission network. PMID- 19131427 TI - A feasibility study of induction pemetrexed plus cisplatin followed by extrapleural pneumonectomy and postoperative hemithoracic radiation for malignant pleural mesothelioma. AB - A prospective multi-institutional study has been commenced in Japan to evaluate the feasibility of induction chemotherapy using pemetrexed plus cisplatin, followed by extrapleural pneumonectomy (EPP) and postoperative hemithoracic radiation in patients with resectable malignant pleural mesothelioma. The study was initiated on May 2008 and 40 patients will be recruited over 3 years. Primary endpoints are macroscopic complete resection rate by EPP and treatment-related mortality for trimodality therapy. Secondary endpoints include treatment completion rate, adverse events, response rate by chemotherapy and 2-year overall and relapse-free survival. PMID- 19131428 TI - Increased gamma-tubulin expression and P16INK4A promoter methylation occur together in preinvasive lesions and carcinomas of the breast. AB - BACKGROUND: Loss of p16(INK4A) due to promoter hypermethylation is correlated with the ability to acquire centrosomal abnormalities in variant human mammary epithelial cells. gamma-Tubulin is a highly conserved component of centrosome in most animal cells and gamma-tubulin protein overexpression could lead to centrosome aberration. MATERIALS AND METHODS: A large series of breast premalignant lesions and carcinoma was analyzed. Real-time quantitative PCR and immunohistochemistry were carried out to measure gamma-tubulin copy numbers and protein expression. MethyLight and immunohistochemistry were carried out to determine p16(INK4A) methylation and protein expression. RESULTS: gamma-Tubulin protein expression was concordant with gene amplification; both of them were found to increase with atypical ductal hyperplasia-carcinoma sequence. The median value and positive rate of p16(INK4a) methylation increased while protein expression displayed a decreasing trend. P16(INK4a) methylation showed a firm association with gamma-tubulin gene amplification. CONCLUSION: gamma-Tubulin gene amplification and the concomitant protein overexpression present not only in invasive carcinoma but also in a significant fraction of atypical hyperplasia and in situ carcinomas. P16(INK4a) methylation and gamma-tubulin gene amplification had a synergistic effect on tumor progression. The synergism might arise as a result of the combined influence that p16(INK4a) and gamma-tubulin have on the G1 S cell cycle checkpoints and centrosome. PMID- 19131429 TI - Controlling an outbreak of MRSA in the neonatal unit: a steep learning curve. AB - Meticillin resistant Staphylococcus aureus (MRSA) can cause serious infections in the newborn. While audit may show that a neonatal unit's main cause of infective morbidity is the coagulase negative staphylococcus, health authorities and politicians fear the implications of MRSA and its impact on the general public. MRSA causes mortality and morbidity in other areas of hospitals in the UK and in many other countries and there is an uneasy acceptance that this is now the established norm. However, MRSA in the neonatal unit carries sensitivities which have a huge impact on the reactions of health authorities, politicians and the press. PMID- 19131430 TI - Do babies with isolated single umbilical artery need routine postnatal renal ultrasonography? AB - BACKGROUND: The presence of isolated single umbilical artery (SUA) in infants has been associated with an increased risk of occult renal malformations. However, the need for routine postnatal renal imaging of such infants, especially in an era of now routine antenatal fetal sonography, is controversial. AIM: To determine the prevalence of significant renal anomalies and the need for routine postnatal renal imaging in infants with isolated SUA. METHODS: Consecutive infants born over 6 years with isolated SUA were offered renal sonography at 4-8 weeks of age. The prevalence of clinically significant renal anomalies in these infants was compared with that detected through routine antenatal fetal scanning and postnatal case findings in a geographically defined control cohort. RESULTS: During the study period, SUA was found in 137 of 33 067 (4.1/1000) live born infants. Infants with isolated SUA (n = 129) were significantly more likely to be preterm and small for gestational age. 122 infants with isolated SUA (95%) underwent renal ultrasonography; only two infants (1.6%, 95% CI 0.20 to 5.5) had clinically significant renal anomalies, a prevalence similar to that in the control cohort (0.4%, 95% CI 0.29 to 0.45; p = 0.74). Four of eight infants with coexistent systemic malformations had abnormal postnatal renal imaging. CONCLUSION: The presence of isolated SUA is associated with increased risk of prematurity and fetal growth restriction. In this largest series of isolated SUA, there was no excess of significant renal malformations among infants with isolated SUA. Postnatal renal ultrasonography is not routinely warranted in such infants. PMID- 19131431 TI - Histological chorioamnionitis and respiratory outcome in preterm infants. AB - A considerable body of human and animal experimental evidence links antenatal inflammation to both accelerated maturation and adverse development of the lung. Initial reports suggest that in preterm infants histological chorioamnionitis is associated with a decreased incidence of respiratory distress syndrome (RDS), while the incidence of bronchopulmonary dysplasia (BPD) is increased. Considerable variation exists in the findings of subsequent human studies, largely dependent on differences in inclusion and exclusion criteria. Taking these differences into account, recent studies generally seem to confirm the effect of chorioamnionitis on RDS incidence, while no effect on BPD is seen. The increased use of antenatal steroids and the diminished effects of secondary pro inflammatory hits seem to explain part of this change. Additional research is needed to explore these complex interactions and their underlying mechanisms, and evaluate the long term pulmonary effects of antenatal inflammation. PMID- 19131432 TI - Randomised crossover trial of four nasal respiratory support systems for apnoea of prematurity in very low birthweight infants. AB - BACKGROUND: Apnoea of prematurity (AOP) is a common problem in preterm infants which can be treated with various modes of nasal continuous positive airway pressure (NCPAP) or nasal intermittent positive pressure ventilation (NIPPV). It is not known which mode of NCPAP or NIPPV is most effective for AOP. OBJECTIVE: To assess the effect of four NCPAP/NIPPV systems on the rate of bradycardias and desaturation events in very low birthweight infants. METHODS: Sixteen infants (mean gestational age at time of study 31 weeks, 10 males) with AOP were enrolled in a randomised controlled trial with a crossover design. The infants were allocated to receive nasal pressure support using four different modes for 6 h each: NIPPV via a conventional ventilator, NIPPV and NCPAP via a variable flow device, and NCPAP delivered via a constant flow underwater bubble system. The primary outcome was the cumulative event rate of bradycardias (< or =80 beats per minute) and desaturation events (< or =80% arterial oxygen saturation), which was obtained from cardio-respiratory recordings. RESULTS: The median event rate was 6.7 per hour with the conventional ventilator in NIPPV mode, and 2.8 and 4.4 per hour with the variable flow device in NCPAP and NIPPV mode, respectively (p value<0.03 for both compared to NIPPV/conventional ventilator). There was no significant difference between the NIPPV/conventional ventilator and the underwater bubble system. CONCLUSION: A variable flow NCPAP device may be more effective in treating AOP in preterm infants than a conventional ventilator in NIPPV mode. It remains unclear whether synchronised NIPPV would be even more effective. PMID- 19131433 TI - Early discharge with tube feeding at home for preterm infants is associated with longer duration of breast feeding. AB - OBJECTIVE: Mothers of preterm infants are more likely to discontinue breast feeding early than mothers of term infants. We evaluated the effect of early discharge with tube feeding of preterm infants under close supervision by paediatric nurse specialists on the duration of breast feeding. DESIGN: Case control study. SETTING: Medium/high-care neonatal unit of a large district general hospital. SUBJECTS: Preterm infants (<37 weeks' gestational age). INTERVENTIONS: Early discharge with tube feeding under close supervision by paediatric nurse specialists or regular follow-up of preterm infants discharged with oral feeding. MAIN OUTCOME MEASURE: Duration of breast feeding assessed by telephone interview 6 months after birth. RESULTS: There were 50 preterm infants in the early discharge group and 78 in the control group. Mothers in the early discharge group continued to breast feed longer than mothers in the control group (log rank test, p = 0.028). Four months after discharge, 63% of preterm infants in the control group were fed formula compared to 36% in the early discharge group (95% CI for difference 9% to 43%, p = 0.04). The relative risk of breast feeding cessation 6 months after birth in the early discharge group compared to the control group was 0.63 (95% CI 0.41 to 0.96). After adjustment for smoking, gestational age and birth weight, this relative risk was 0.67 (95% CI 0.43 to 1.05). CONCLUSIONS: Close supervision and follow-up by paediatric nurse specialists of preterm infants discharged early with tube feeding appears to increase duration of breast feeding. A randomised controlled trial to confirm these findings is warranted. PMID- 19131434 TI - Robust production and passaging of infectious HPV in squamous epithelium of primary human keratinocytes. AB - Using Cre-loxP-mediated recombination, we established a highly efficient and reproducible system that generates autonomous HPV-18 genomes in primary human keratinocytes (PHKs), the organotypic raft cultures of which recapitulated a robust productive program. While E7 promoted S-phase re-entry in numerous suprabasal differentiated cells, HPV DNA unexpectedly amplified following a prolonged G2 arrest in mid- and upper spinous cells. As viral DNA levels intensified, E7 activity diminished and then extinguished. These cells then exited the cell cycle to undergo virion morphogenesis. High titers of progeny virus generated an indistinguishable productive infection in naive PHK raft cultures as before, never before achieved until now. An immortalization-defective HPV-18 E6 mutant genome was also characterized for the first time. Numerous cells accumulated p53 protein, without inducing apoptosis, but the productive program was severely curtailed. Complementation of mutant genomes by E6-expressing retrovirus restored proper degradation of p53 as well as viral DNA amplification and L1 production. This system will be invaluable for HPV genetic dissection and serves as a faithful ex vivo model for investigating infections and interventions. PMID- 19131435 TI - Drosophila PTB promotes formation of high-order RNP particles and represses oskar translation. AB - Local translation of asymmetrically enriched mRNAs is a powerful mechanism for functional polarization of the cell. In Drosophila, exclusive accumulation of Oskar protein at the posterior pole of the oocyte is essential for development of the future embryo. This is achieved by the formation of a dynamic oskar ribonucleoprotein (RNP) complex regulating the transport of oskar mRNA, its translational repression while unlocalized, and its translational activation upon arrival at the posterior pole. We identified the nucleo-cytoplasmic shuttling protein PTB (polypyrimidine tract-binding protein)/hnRNP I as a new factor associating with the oskar RNP in vivo. While PTB function is largely dispensable for oskar mRNA transport, it is necessary for translational repression of the localizing mRNA. Unexpectedly, a cytoplasmic form of PTB can associate with oskar mRNA and repress its translation, suggesting that nuclear recruitment of PTB to oskar complexes is not required for its regulatory function. Furthermore, PTB binds directly to multiple sites along the oskar 3' untranslated region and mediates assembly of high-order complexes containing multiple oskar RNA molecules in vivo. Thus, PTB is a key structural component of oskar RNP complexes that dually controls formation of high-order RNP particles and translational silencing. PMID- 19131436 TI - Prefrontostriatal circuitry regulates effort-related decision making. AB - The anterior cingulate cortex (ACC), the basolateral amygdala (BLA), and the dopamine in the nucleus accumbens (NAc) are part of a neural system that is critically involved in making decisions on how much effort to invest for rewards. In the present study, we sought to identify functional interactions between ACC and NAc regulating effort-based decision making. Rats were tested in a T-maze cost-benefit task in which they could either choose to climb a barrier to obtain a large reward (LR) in one arm or a small reward in the other arm without a barrier. Experiment 1 revealed that bilateral excitotoxic lesions of the core subregion of the NAc impaired effort-based decision making, that is, reduced the preference for the high effort-LR option when having the choice to obtain a low reward with little effort. Experiment 2 showed that disconnection of the ACC and NAc core using an asymmetrical excitotoxic lesion procedure impaired effort-based decision making. The present data provide evidence that effort-based decision making is governed by an interconnected neural system that requires serial information transfer between ACC and NAc core. PMID- 19131437 TI - Excitatory GABAergic activation of cortical dividing glial cells. AB - Adult neocortex contains dividing satellite glia population even though their characteristics and functions have still remained unknown. Nestin(+)/NG2(+) cells as major fraction of dividing glial cells express bicuculline-sensitive gamma aminobutyric acid A (GABA(A)) receptors and receive GABAergic inputs. Due to their high [Cl(-)](i), GABAergic activation depolarized the cells and then induced Ca(2+) influx into them. To assess an effect of this GABAergic excitation, we looked for the expression of neurotrophic factors. Among them, we detected the expression of brain-derived neurotrophic factor (BDNF) on the cells. The level of BDNF expression was elevated after cortical ischemia, and this elevation was blocked by bumetanide, an inhibitor for NKCC1 that blocks the GABAergic depolarization. Furthermore, performing a modified adhesive removal test, we observed that the treatment of bumetanide significantly attenuated the recovery in somatosensory dysfunction. Our results may shed a light on satellite glia population in the cortex and imply their roles in the functional recovery after ischemic injuries. PMID- 19131438 TI - Acute lymphoblastic leukemia--on the wings of IKAROS. PMID- 19131439 TI - Simulation of congenital heart defects: a novel way of training in echocardiography. AB - BACKGROUND: Echocardiography is one of the most important diagnostic imaging modalities in paediatric cardiology. Owing to the large number of lesions, achieving expertise often requires years of training. Echocardiography is still taught using the apprenticeship model, which is time- and personnel consuming. OBJECTIVES: To extend the echocardiography simulator EchoCom to enable simulation of congenital heart lesions and validate it for training in paediatric echocardiography. METHODS: The simulator consists of a life-size manikin, a dummy transducer with attached three-dimensional (3D) tracking system and a computer application. Transthoracic real-time (RT) 3D echocardiographic datasets were collected and embedded into the simulator. Two-dimensional images were calculated and resliced from these datasets according to the position of the tracking sensor. Ten RT 3D datasets of congenital heart lesions were selected for validation. Datasets were blinded and without additional information presented to 43 participants who were stratified according to their expertise (12 experts, 16 intermediates, 15 beginners). Participants were asked to list the relevant findings and make a diagnosis. Construct validation was tested comparing diagnostic performance for each group. Face and content validation were tested using a standardised questionnaire. RESULTS: Participants judged the simulator as realistic and useful. The main drawback was the adult size of the manikin. The diagnostic performance of each group differed significantly proving construct validity. CONCLUSIONS: According to this validation the prototype simulator could make a significant contribution to training in the use of echocardiography in congenital heart disease. PMID- 19131440 TI - Acute aortic syndrome: a new look at an old conundrum. AB - The term acute aortic syndrome (AAS), coined several years ago, is now widely recognised. In the light of new findings in aortic pathology and in an era when modern imaging techniques are widely available and interventional management of AAS is increasing, some morphological and diagnostic aspects of acute aortic pathology have been examined and the syndrome updated. This article provides a new, comprehensive overview of the pathology, diagnosis, evolution and management of patients with AAS. As acute aortic disease is the most common fatal condition in patients with chest pain, prompt recognition and treatment is of paramount importance. PMID- 19131441 TI - Cardiovascular risk: who should we treat, and how much should we stratify? PMID- 19131442 TI - n-3 Polyunsaturated fatty acids are still underappreciated and underused post myocardial infarction. PMID- 19131443 TI - Relationship between sleep apnoea and mortality in patients with ischaemic heart failure. AB - OBJECTIVE: To determine whether the influence of sleep apnoea (SA) on the risk of death differs in patients with ischaemic and in those with non-ischaemic heart failure (HF). DESIGN: Prospective observational study. PATIENTS: Consecutive patients with HF with left ventricular ejection fraction < or =45% newly referred to the HF clinic between 1 September 1997 and 1 December 2004. MAIN OUTCOME MEASURES: Patients underwent sleep studies and were divided into those with moderate to severe SA (apnoea-hypopnoea index > or =15/h of sleep) and those with mild to no SA (apnoea-hypopnoea index <15/h of sleep). They were followed up for a mean of 32 months to determine all-cause mortality rate. RESULTS: Of 193 patients, 34 (18%) died. In the ischaemic group, mortality risk adjusted for confounding factors was significantly higher in those with SA than in those without it (18.9 vs 4.6 deaths/100 patient-years, hazards ratio (HR) = 3.03, 95% CI 1.04 to 8.84, p = 0.043). In contrast, in the non-ischaemic HF group, there was no difference in adjusted mortality risk between those with, and those without, SA (3.9 vs 4.0 deaths/100 patient-years, p = 0.929). CONCLUSIONS: In patients with HF, the presence of SA is independently associated with an increased risk of death in those with ischaemic, but not in those with non ischaemic, aetiology. These findings suggest that patients with ischaemic cardiomyopathy are more susceptible to the adverse haemodynamic, autonomic and inflammatory consequences of SA than are those with non-ischaemic cardiomyopathy. PMID- 19131444 TI - Challenges and opportunities for primary, secondary, and tertiary prevention of Chagas' disease. AB - A century after its discovery, Chagas' disease still represents a major public health challenge in Latin America. Moreover, because of growing population movements, an increasing number of cases of imported Chagas' disease have now been detected in non-endemic areas, such as North America and some European countries. This parasitic zoonosis, caused by Trypanosoma cruzi, is transmitted to humans by infected Triatominae insects, or occasionally by non-vectorial mechanisms, such as blood transfusion, mother to fetus, or oral ingestion of materials contaminated with parasites. Following the acute phase of the infection, untreated individuals enter a chronic phase that is initially asymptomatic or clinically unapparent. Usually, a few decades later, 40-50% of patients develop progressive cardiomyopathy and/or motility disturbances of the oesophagus and colon. In the last decades several interventions targeting primary, secondary and tertiary prevention of Chagas' disease have been attempted. While control of both vectorial and blood transfusion transmission of T cruzi (primary prevention) has been successful in many regions of Latin America, early detection and aetiological treatment of asymptomatic subjects with Chagas' disease (secondary prevention) have been largely underutilised. At the same time, in patients with established chronic disease, several pharmacological and non-pharmacological interventions are currently available and have been increasingly used with the intention of preventing or delaying complications of the disease (tertiary prevention). In this review we discuss in detail each of these issues. PMID- 19131445 TI - The eye: window to the soul or a mirror of systemic health? PMID- 19131446 TI - States, rates and traits: prognosis research and chronic stable angina. PMID- 19131447 TI - Sex differences and predictors of objective cough frequency in chronic cough. AB - BACKGROUND: Women are consistently over-represented in specialist cough clinics and known to have a more sensitive cough reflex than men. Whether female sex and other patient characteristics are associated with higher cough rates is not known. A study was conducted to determine the predictors of objective cough frequency in patients presenting to a tertiary referral clinic with chronic cough. METHODS: 100 subjects (65 women) of mean (SD) age 55.8 (11.0) years and median cough duration 4 years (IQR 2.0-10.0) with unexplained chronic cough completed flow-volume loops (mean (SD) forced expiratory volume in 1 s 103 (15.2)% predicted; forced expiratory flow (FEF(50)) 68.8 (24.1)% predicted), methacholine challenge (42% positive), citric acid cough reflex sensitivity (C5; 0.12 M (IQR 0.06-0.50)) and the Leicester Cough Questionnaire. 24-h ambulatory cough monitoring was performed in 86 subjects; manually counted coughs were quantified as the number of explosive cough sounds per hour. RESULTS: Women coughed significantly more than men (geometric mean 16.6 coughs/h (95% CI 13.1 to 21.0) vs 9.4 coughs/h (95% CI 6.4 to 13.9), p = 0.01)). The cough reflex was also more sensitive in women than in men (median logC5 -0.9 M vs -0.6 M, p = 0.002), but cough-related quality of life was similar in women and men (12.0 (3.6) and 12.2 (3.2), respectively, p = 0.76). Linear regression analysis showed that 38.6% of the variation in cough rate was predicted by sex (p = 0.01), logC5 (p<0.001) and age (p = 0.002) but not lung function or bronchial hyper-reactivity. CONCLUSIONS: Ambulatory objective cough monitoring provides novel insights into factors modulating chronic cough. These findings suggest that effects of sex and age must be taken into account in the study of cough and when designing clinical trials testing novel antitussive agents. PMID- 19131448 TI - Biomarkers improve mortality prediction by prognostic scales in community acquired pneumonia. AB - BACKGROUND: Prognostic scales provide a useful tool to predict mortality in community-acquired pneumonia (CAP). However, the inflammatory response of the host, crucial in resolution and outcome, is not included in the prognostic scales. METHODS: The aim of this study was to investigate whether information about the initial inflammatory cytokine profile and markers increases the accuracy of prognostic scales to predict 30-day mortality. To this aim, a prospective cohort study in two tertiary care hospitals was designed. Procalcitonin (PCT), C-reactive protein (CRP) and the systemic cytokines tumour necrosis factor alpha (TNFalpha) and interleukins IL6, IL8 and IL10 were measured at admission. Initial severity was assessed by PSI (Pneumonia Severity Index), CURB65 (Confusion, Urea nitrogen, Respiratory rate, Blood pressure, > or = 65 years of age) and CRB65 (Confusion, Respiratory rate, Blood pressure, > or = 65 years of age) scales. A total of 453 hospitalised CAP patients were included. RESULTS: The 36 patients who died (7.8%) had significantly increased levels of IL6, IL8, PCT and CRP. In regression logistic analyses, high levels of CRP and IL6 showed an independent predictive value for predicting 30-day mortality, after adjustment for prognostic scales. Adding CRP to PSI significantly increased the area under the receiver operating characteristic curve (AUC) from 0.80 to 0.85, that of CURB65 from 0.82 to 0.85 and that of CRB65 from 0.79 to 0.85. Adding IL6 or PCT values to CRP did not significantly increase the AUC of any scale. When using two scales (PSI and CURB65/CRB65) and CRP simultaneously the AUC was 0.88. CONCLUSIONS: Adding CRP levels to PSI, CURB65 and CRB65 scales improves the 30 day mortality prediction. The highest predictive value is reached with a combination of two scales and CRP. Further validation of that improvement is needed. PMID- 19131449 TI - Risk factors for complicated parapneumonic effusion and empyema on presentation to hospital with community-acquired pneumonia. AB - BACKGROUND: The aim of this study was to identify key factors on admission predicting the development of complicated parapneumonic effusion or empyema in patients admitted with community-acquired pneumonia. METHODS: A prospective observational study of patients admitted with community-acquired pneumonia in NHS Lothian, UK, was conducted. Multivariate regression analyses were used to evaluate factors that could predict the development of complicated parapneumonic effusion or empyema, including admission demographics, clinical features, laboratory tests and pneumonia-specific (Pneumonia Severity Index (PSI), CURB65 (New onset confusion, urea >7 mmol/l, Respiratory rate > or = 30 breaths/min, Systolic blood pressure < 90 mm Hg and/or diastolic blood pressure < or = 60 mm Hg and age > or = 65 years) and CRB65 (New onset confusion, Respiratory rate > or = 30 breaths/min, Systolic blood pressure <90 mm Hg and/or diastolic blood pressure < or = 60 mm Hg and age > or = 65 years)) and generic sepsis scoring systems (APACHE II (Acute Physiology and Chronic Health Evaluation II), SEWS (standardised early warning score) and systemic inflammatory response syndrome (SIRS)). RESULTS: 1269 patients were included in the study and 92 patients (7.2%) developed complicated parapneumonic effusion or empyema. The pneumonia-specific and generic sepsis scoring systems had no value in predicting complicated parapneumonic effusion or empyema. Multivariate logistic regression identified albumin <30 g/l adjusted odds ratio (AOR) 4.55 (95% CI 2.45 to 8.45, p < 0.0001), sodium <130 mmol/l AOR 2.70 (1.55 to 4.70, p = 0.0005), platelet count >400 x 10(9)/l AOR 4.09 (2.21 to 7.54, p < 0.0001), C-reactive protein >100 mg/l AOR 15.7 (3.69 to 66.9, p < 0.0001) and a history of alcohol abuse AOR 4.28 (1.87 to 9.82, p = 0.0006) or intravenous drug use AOR 2.82 (1.09 to 7.30, p = 0.03) as independently associated with development of complicated parapneumonic effusion or empyema. A history of chronic obstructive pulmonary disease was associated with decreased risk, AOR 0.18 (0.06 to 0.53, p = 0.002). A 6-point scoring system using these combined variables had good discriminatory value: area under the receiver operator characteristic curve (AUC) 0.84 (95% CI 0.81 to 0.86, p < 0.0001). CONCLUSION: This study has identified seven clinical factors predicting the development of complicated parapneumonic effusion or empyema. Independent validation is needed. PMID- 19131450 TI - Heterogeneity of asthma according to blood inflammatory patterns. AB - RATIONALE: There is increasing interest regarding asthma heterogeneity in relation to inflammatory patterns. OBJECTIVES: To assess phenotypic characteristics, in particular clinical presentation of the disease, in 381 well characterised adults with asthma from the French Epidemiological study on the Genetics and Environment of Asthma (EGEA) according to their blood inflammatory pattern. METHODS: Four blood inflammatory patterns were defined according to eosinophil (EOS) and neutrophil (NEU) count cut-off points. Samples with > or =250 EOS/mm(3) were classified as EOS(hi) and those with > or =5000 NEU/mm(3) as NEU(hi). Clinical characteristics include typical asthma and chronic obstructive pulmonary disease (COPD)-like symptoms, as well as composite quantitative scores addressing the activity of the disease. RESULTS: A substantial number of those with asthma (56.2%) had the EOS(lo) pattern (<250 EOS/mm(3)). Patients with asthma who had the EOS(hi) pattern had higher immunoglobulin E (IgE), a lower forced expiratory volume in 1 s (FEV(1)) and presented a more active asthma than those with the EOS(lo) pattern. Among those with the EOS(lo) pattern, neutrophil inflammation (NEU(hi)) was related to a less frequent positive skin prick test response (OR 0.44, 95% CI 0.20 to 0.96). Among those with the EOS(hi) pattern, neutrophil inflammation did not explain current asthma or asthma activity, and was significantly related to nocturnal symptoms (OR 5.21, 95% CI 1.44 to 18.8) independently of age, sex, smoking and inhaled corticosteroid treatment. In non smokers with asthma, COPD-like symptoms, in particular chronic phlegm, were more frequent in those with neutrophil inflammation, independent of eosinophil inflammation (OR 2.35, 95% CI 1.08 to 5.10). CONCLUSIONS: Besides eosinophilia, neutrophil inflammation assessed in the blood is related to specific characteristics of asthma. Considering simultaneously neutrophilic and eosinophilic inflammation may contribute to help to disentangle this complex disease. PMID- 19131451 TI - Novel site-specific mast cell subpopulations in the human lung. AB - BACKGROUND: Lung mast cells are stereotypically divided into connective tissue (MC(TC)) and mucosal (MC(T)) mast cells. This study tests the hypothesis that each of these subtypes can be divided further into site-specific populations created by the microenvironment within each anatomical lung compartment. METHODS: Surgical resections and bronchial and transbronchial biopsies from non-smoking individuals were obtained to study mast cells under non-inflamed conditions. Morphometric and molecular characteristics of mast cell populations were investigated in multiple lung structures by immunohistochemistry and electron microscopy. RESULTS: MC(T) and MC(TC) coexisted in all compartments, with MC(T) being the prevailing type in bronchi, bronchioles and the alveolar parenchyma and MC(TC) being more abundant in pulmonary vessels and the pleura. Each of the MC(TC) and MC(T) phenotypes could be further differentiated into site-specific populations. MC(TC) were significantly larger in pulmonary vessels than in small airway walls, while the reverse was observed for MC(T). Within each MC(TC) and MC(T) population there were also distinct site-specific expression patterns of the IgE receptor, interleukin-9 receptor, renin, histidine decarboxylase, vascular endothelial growth factor, fibroblast growth factor, 5-lipoxygenase and leukotriene C4 synthase (eg, bronchial MC(T) consistently expressed more histidine decarboxylase than alveolar MC(T)). Renin content was high in vascular MC(TC) but markedly lower in MC(TC) in other compartments. For both MC(TC) and MC(T), the IgE receptor was highly expressed in conducting airways but virtually absent in alveolar parenchyma. CONCLUSIONS: These findings demonstrate novel site specific subpopulations of lung MC(TC) and MC(T) at baseline conditions. This observation may have important implications in the future exploration of mast cells in a number of pulmonary diseases. PMID- 19131452 TI - Genetic association study for RSV bronchiolitis in infancy at the 5q31 cytokine cluster. AB - BACKGROUND: The pathophysiological basis of severe respiratory syncytial virus (RSV) bronchiolitis in infancy is poorly understood and has hindered vaccine development. Studies implicate the cell-mediated immune response in the pathogenesis of the disease. A recent twin study estimated a heritable contribution of 22% to RSV bronchiolitis. Genetic epidemiology provides a new approach to identifying important immune determinants of disease severity. METHODS: A comprehensive high-density gene-region association study for severe RSV bronchiolitis in infancy at 5q31 across 11 genes including the Th2-cytokine cluster was performed. A haplotype tagging approach was used to analyse genetic variation at 113 single nucleotide polymorphisms (SNPs) in 780 independent cases and 1045 controls. The study had sufficient power to detect small effects, perform extensive haplotype analysis and analyse both a principal phenotype and a refined age-limited phenotype enriched for first-exposure RSV infection. RESULTS: SNP associations were found at IL4 and a highly significant risk haplotype was identified across IL13 CNS-1 and IL4 (odds ratio 1.69, p<0.0001), present in both case-control and family-based analyses. All associations were strongest for a phenotype limited to <6 months of age, implicating this locus in primary RSV disease. The same risk haplotype has previously been shown to be associated with increased IL13 expression. CONCLUSIONS: A haplotype at IL13-1L4, which is associated with increased IL13 production, confers an increased risk of severe primary RSV bronchiolitis in early infancy. This study, together with previous studies implicating the same locus in atopic sensitisation, suggests that primary RSV bronchiolitis and atopy share a genetic contribution at the IL13-IL4 locus. PMID- 19131453 TI - Economic cost of tobacco use in India, 2004. AB - OBJECTIVE: To estimate the tobacco-attributable costs of diseases separately for smoked and smokeless tobacco use in India. METHODS: The prevalence-based attributable-risk approach was used to estimate the economic cost of tobacco using healthcare expenditure data from the National Sample Survey, a nationally representative household sample survey conducted in India in 2004. Four major categories of tobacco-related disease-tuberculosis, respiratory diseases, cardiovascular diseases and neoplasms-were considered. RESULTS: Direct medical costs of treating tobacco related diseases in India amounted to $907 million for smoked tobacco and $285 million for smokeless tobacco. The indirect morbidity costs of tobacco use, which includes the cost of caregivers and value of work loss due to illness, amounted to $398 million for smoked tobacco and $104 million for smokeless tobacco. The total economic cost of tobacco use amounted to $1.7 billion. Tuberculosis accounted for 18% of tobacco-related costs ($311 million) in India. Of the total cost of tobacco, 88% was attributed to men. CONCLUSIONS: The cost of tobacco use was many times more than the expenditures on tobacco control by the government of India and about 16% more than the total tax revenue from tobacco. The tobacco-attributable cost of tuberculosis was three times higher than the expenditure on tuberculosis control in India. The economic costs estimated here do not include the costs of premature mortality from tobacco use, which is known to comprise roughly 50% to 80% of the total economic cost of tobacco in many countries. PMID- 19131454 TI - Using a quitline plus low-cost nicotine replacement therapy to help disadvantaged smokers to quit. AB - OBJECTIVES: To trial an intervention in a real-life setting to motivate low income smokers to try to quit. The intervention under trial was the addition of subsidised nicotine replacement therapy (NRT) to a standard population quitline service. DESIGN: Participants were low-income smokers, recruited "cold" via either a letter in the mail or a flyer inserted in a local newspaper. The intervention group received the usual service of multisession counselling from the quitline plus access to heavily subsidised NRT. A comparison group received the usual quitline service only. Participants were followed up at 3, 6, and 12 months. Trial participants were also compared with a sample of general callers to the quitline. RESULTS: The offer of subsidised NRT recruited more than twice as many low-income smokers than the offer of the cessation service alone (intervention group n = 1000; comparison group n = 377). 63% were first-time callers to the quitline. Intervention respondents showed higher levels of nicotine dependence than comparison group respondents. Comparisons of quitting data were confounded by the differences in the respondents at baseline. 73.5% of smokers in the intervention group tried to quit compared to 61.0% in the comparison group. Unadjusted quit rates were higher in the intervention group than in the comparison group at 3 months and 6 months but not at 12 months. CONCLUSIONS: Disadvantaged smokers were easily engaged to call the quitline, particularly when offered subsidised NRT. Disadvantaged smokers using the quitline, with and without subsidised NRT, achieved cessation outcomes comparable to other studies of "mainstream" smokers. PMID- 19131455 TI - Predicting smokers' non-compliance with smoking restrictions in public places. AB - OBJECTIVE: The present study aimed to identify the predictors of non-compliance with smoking restrictions among Greek college student smokers. Differences in attitudes to smoking bans and tobacco control policies between current smokers and non-smokers were also examined. METHODS: Data were collected from college students (n = 229, mean (SD) age 21.27 (3.15) years). Measures included tobacco dependence, attitudes to tobacco control policies, outcome expectancy and normative beliefs. RESULTS: The majority of current smokers reported non compliance with existing restrictions. Logistic regression analysis showed that non-compliance was significantly predicted by (less) anticipated regret from tobacco-related health harm, tobacco dependence and perceived prevalence and social acceptability of smoking. Analysis of variance indicated that current non smokers held significantly more positive attitudes to smoking bans and tobacco control policies than current smokers. CONCLUSIONS: Smoking was highly prevalent, and more than half of current smokers reported not complying with existing regulations in public places. Smokers' attitudes to smoking bans and tobacco control policies did not have an effect on compliance behaviour. Future policies to promote compliance with smoking restrictions in Greece should target health related anticipated regret, and perceived social norms. PMID- 19131456 TI - The prevalence of household second-hand smoke exposure and its correlated factors in six counties of China. AB - OBJECTIVE: To study the prevalence of, and discuss factors contributing to, household second-hand smoke exposure in six counties in China, providing scientific support for the need to establish tobacco control measures in these areas. METHODS: A cross-sectional survey was performed. Investigators conducted face-to-face interviews using a standardised questionnaire to collect information on demographics, passive smoking behaviours and knowledge, and attitudes towards tobacco control. The setting was six counties from the three provinces: Mianzhu and Xichong counties in Sichuan Province; Anyi and Hukou counties in Jiangxi Province; and Xinan and Yanshi counties in Henan Province. A total of 8142 non smokers (aged 18-69) in 2004 were included in the data analysis. Household second hand smoke exposure rate as defined as the proportion of household passive smokers in the non-smoker population was used as the measure of household second hand smoke exposure. RESULTS: The analysis of 8142 non-smokers revealed that, in these selected counties, the household second-hand smoke exposure rate was 48.3%. Respondents had positive attitudes towards tobacco control. Of 6972 respondents, 84.4% supported all the three tobacco control policies (banning smoking in public places, banning the selling of cigarettes to minors, banning all cigarette advertisements). In 3165 families with smokers, 87.2% of respondents reported that smokers would smoke in front of them. In 2124 families with smokers and children, 76.5% of respondents reported that smokers would smoke in front of children. As many as 42.1% of non-smokers would offer cigarettes to their guests, and only 46.8% of respondents would ask smokers to smoke outdoors. Only 6.3% of families completely forbade smoking at home. Multivariate logistic regression analysis revealed high second-hand smoke exposure for the following demographic groups: Jiangxi Province inhabitants, females, those with low education level, farmers and married respondents. CONCLUSIONS: Household second-hand smoke exposure rates in the selected counties were high. A high percentage of respondents reported that smokers would smoke in front of them and children. The pressure from non-smokers against smoking was relatively low, although offering cigarette was prevalent. Households that were completely smoking-free were rare, Further studies on these correlated factors could help us establish effective measures to reduce household second-hand smoke exposure. PMID- 19131457 TI - Misconceptions about diabetes and its management among low-income minorities with diabetes. AB - OBJECTIVE: To determine diabetic patients' knowledge and beliefs about the disease and medications that could hinder optimal disease management. RESEARCH DESIGN AND METHODS: A cross-sectional survey of 151 type 2 diabetic patients characterizing diabetes knowledge and beliefs about the disease and medications was conducted. RESULTS: Mean diabetes duration was 13 years. Over half of the patients (56%) believed that normal glucose is G genotype groups. Subjects with the SNP 45TT genotype showed increased adiponectin levels and decreased HOMA-IR indexes. Haplotype analysis revealed that homozygous carriers of the TG haplotype (45TT and 276GG) and heterozygous carriers of the TG haplotype (TG/X) showed a reduction in the HOMA-IR index after adjustment for baseline levels. Significant differences were observed in changes in HOMA-IR indexes and adiponectin concentrations according to the 45-276 TG haplotype in overweight-obese, but not in normal-weight subjects: the greatest decrease in HOMA-IR indexes and the greatest increase in adiponectin levels were shown in overweight-obese subjects with the TG/TG haplotype. CONCLUSIONS: ADIPOQ genetic variants can affect circulating adiponectin levels and insulin resistance indexes in subjects with IFG or newly diagnosed type 2 diabetes in response to dietary intervention. PMID- 19131461 TI - Combined measurement of fasting plasma glucose and A1C is effective for the prediction of type 2 diabetes: the Kansai Healthcare Study. AB - OBJECTIVE: We prospectively assessed whether the combined measurements of fasting plasma glucose (FPG) and A1C were effective for predicting type 2 diabetes. RESEARCH DESIGN AND METHODS: Study participants included 6,736 nondiabetic Japanese men aged 40-55 years. Type 2 diabetes was diagnosed in those who had an FPG >or=126 mg/dl or who were being treated with an oral antidiabetic agent or insulin. The models including FPG, A1C, and both were compared using the area under the receiver operating characteristic (AUROC) curves. RESULTS: During the 4 year follow-up period, we confirmed 659 diabetes cases. In multivariate analysis, both FPG and A1C were independently associated with the risk of type 2 diabetes. The model including both FPG and A1C had a greater AUROC curve than that including FPG alone (0.853 vs. 0.818; P < 0.001) or A1C alone (0.853 vs. 0.771; P < 0.001). CONCLUSIONS: The combined measurement of FPG and A1C was effective for predicting type 2 diabetes. PMID- 19131460 TI - Incremental value of the pancreas allograft to the survival of simultaneous pancreas-kidney transplant recipients. AB - OBJECTIVE: To quantify the incremental survival benefit of the pancreas allograft in simultaneous pancreas-kidney (SPK) transplant recipients. RESEARCH DESIGN AND METHODS: Data from the national transplant database from 2000 to 2007 were analyzed. SPK recipients who had functioning allografts to 1-year post transplant (n = 3,304) were compared with those who had failure of the renal (n = 233) or pancreatic (n = 112) graft. The main outcome was a projection of 10 life-years of patient survival beyond the first transplant anniversary. RESULTS: Recipients with function of both organs accrued 9.4 life-years following transplantation. Projected survival in patients with kidney failure was reduced to 2.5 life-years. Pancreas failure reduced predicted survival to 8 life-years. Renal allograft failure impacts life expectancy significantly (adjusted hazard ratio [aHR] 12.13). However, pancreas allograft failure was also associated with reduced survival (aHR 2.62). CONCLUSIONS: Although the majority of the survival benefit of SPK transplant is due to the renal transplant, pancreas allograft function does contribute to patient survival. PMID- 19131462 TI - Antidiabetic action of bezafibrate in a large observational database. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that bezafibrate, an approved fibrate, can prevent or delay type 2 diabetes. RESEARCH DESIGN AND METHODS: This was a retrospective cohort study using data from routine medical practice in the U.K., as captured by the General Practice Research Database (GPRD). Individuals chronically exposed to bezafibrate were compared with individuals chronically exposed to other fibrates. Hazard ratios (HRs) for incident type 2 diabetes were calculated using a Cox proportional hazards model. A post hoc analysis was used to examine the effect of bezafibrate on progression to use of oral antidiabetic medications or insulin in individuals with diabetes at baseline. RESULTS: Bezafibrate users had a lower hazard for incident diabetes than users of other fibrates (HR 0.66 [95% CI 0.53-0.81]). This effect became stronger with increasing duration of therapy. Post hoc analysis of the effect of bezafibrate on progression of preexisting diabetes also showed a lower hazard for progression to use of antidiabetic medication (0.54 [0.38-0.76]) or progression to use of insulin (0.78 [0.55-1.10]). CONCLUSIONS: Bezafibrate appears to have clinically important antidiabetic properties. Randomized controlled trials should be considered to assess the utility of bezafibrate in treating patients with diabetes or in preventing diabetes in high-risk patients. PMID- 19131463 TI - Subclinical inflammation and diabetic polyneuropathy: MONICA/KORA Survey F3 (Augsburg, Germany). AB - OBJECTIVE: Subclinical inflammation represents a risk factor of type 2 diabetes and several diabetes complications, but data on diabetic neuropathies are scarce. Therefore, we investigated whether circulating concentrations of acute-phase proteins, cytokines, and chemokines differ among diabetic patients with or without diabetic polyneuropathy. RESEARCH DESIGN AND METHODS: We measured 10 markers of subclinical inflammation in 227 type 2 diabetic patients with diabetic polyneuropathy who participated in the population-based MONICA/KORA Survey F3 (2004-2005; Augsburg, Germany). Diabetic polyneuropathy was diagnosed using the Michigan Neuropathy Screening Instrument (MNSI). RESULTS: After adjustment for multiple confounders, high levels of C-reactive protein and interleukin (IL)-6 were most consistently associated with diabetic polyneuropathy, high MNSI score, and specific neuropathic deficits, whereas some inverse associations were seen for IL-18. CONCLUSIONS: This study shows that subclinical inflammation is associated with diabetic polyneuropathy and neuropathic impairments. This association appears rather specific because only certain immune mediators and impairments are involved. PMID- 19131464 TI - Changes in albumin excretion in the diabetes prevention program. AB - OBJECTIVE: Increased urinary albumin excretion rates have been linked to nephropathy and macrovascular disease. We here describe the baseline prevalence and effect of Diabetes Prevention Program (DPP) interventions on the development and reversal of elevated albumin excretion. RESEARCH DESIGN AND METHODS: Urine albumin-to-creatinine ratios (ACRs) were calculated from untimed urine collections. Analyses compared participants by treatment group, diabetes and hypertension status, and use of ACE inhibitors or angiotensin II receptor blockers (ARBs). RESULTS: Elevated ACR levels (>or=30 mg/g creatinine) were present at baseline in 198 (6.2%) of 3,188 participants: placebo 5.3%, metformin 6.5%, and intensive lifestyle (ILS) 6.8%. Of the 2,802 with ACR measurements at baseline and at the end of the study, the percentage with elevated levels declined (incident and regression) from 6.2 to 6.1%, with no significant differences between the groups even with adjustment for ACE inhibitor and ARB use. The odds of developing an elevated ACR were 59% higher for a participant who developed diabetes compared with one who did not. CONCLUSIONS: At entry into the DPP, an elevated ACR was present in 6.2%. Despite the marked decrease in progression to diabetes and the improvement in insulin resistance and other cardiovascular risk markers in the ILS and metformin groups, there was no improvement in ACR, on average, in those two groups. However, the frequency of an elevated ACR was higher in participants who developed diabetes. An increased ACR may have multiple causes, thus obscuring the improvements that might have been expected with the reduction in insulin resistance seen in the DPP. PMID- 19131465 TI - Admission hyperglycemia predicts a worse outcome in stroke patients treated with intravenous thrombolysis. AB - OBJECTIVE: Admission hyperglycemia has been associated with worse outcomes in ischemic stroke. We hypothesized that hyperglycemia (glucose >8.0 mmol/l) in the hyperacute phase would be independently associated with increased mortality, symptomatic intracerebral hemorrhage (SICH), and poor functional status at 90 days in stroke patients treated with intravenous tissue plasminogen activator (IV tPA). RESEARCH DESIGN AND METHODS: Using data from the prospective, multicenter Canadian Alteplase for Stroke Effectiveness Study (CASES), the association between admission glucose >8.0 mmol/l and mortality, SICH, and poor functional status at 90 days (modified Rankin Scale >1) was examined. Similar analyses examining glucose as a continuous measure were conducted. RESULTS: Of 1,098 patients, 296 (27%) had admission hyperglycemia, including 18% of those without diabetes and 70% of those with diabetes. After multivariable logistic regression, admission hyperglycemia was found to be independently associated with increased risk of death (adjusted risk ratio 1.5 [95% CI 1.2-1.9]), SICH (1.69 [0.95 3.00]), and a decreased probability of a favorable outcome at 90 days (0.7 [0.5 0.9]). An incremental risk of death and SICH and unfavorable 90-day outcomes was observed with increasing admission glucose. This observation held true for patients with and without diabetes. CONCLUSIONS: In this cohort of IV-tPA-treated stroke patients, admission hyperglycemia was independently associated with increased risk of death, SICH, and poor functional status at 90 days. Treatment trials continue to be urgently needed to determine whether this is a modifiable risk factor for poor outcome. PMID- 19131466 TI - Alanine aminotransferase, gamma-glutamyltransferase, and incident diabetes: the British Women's Heart and Health Study and meta-analysis. AB - OBJECTIVE: To estimate and compare associations of alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) with incident diabetes. RESEARCH DESIGN AND METHODS: ALT and GGT were studied as determinants of diabetes in the British Women's Heart and Health Study, a cohort of 4,286 women 60-79 years old (median follow-up 7.3 years). A systematic review and a meta-analysis of 21 prospective, population-based studies of ultrasonography, which diagnosed nonalcoholic fatty liver disease (NAFLD), ALT, and GGT as determinants of diabetes, were conducted, and associations of ALT and GGT with diabetes were compared. RESULTS: Ultrasonography-diagnosed NAFLD was associated with more than a doubling in the risk of incident diabetes (three studies). ALT and GGT both predicted diabetes. The fully adjusted hazard ratio (HR) for diabetes per increase in one unit of logged ALT was 1.83 (95% CI 1.57-2.14, I(2) = 8%) and for GGT was 1.92 (1.66 2.21, I(2) = 55%). To directly compare ALT and GGT as determinants of diabetes, the fully adjusted risk of diabetes in the top versus bottom fourth of the ALT and GGT distributions was estimated using data from studies that included results for both markers. For ALT, the HR was 2.02 (1.59-2.58, I(2) = 27%), and for GGT the HR was 2.94 (1.98-3.88, I(2) = 20%), suggesting that GGT may be a better predictor (P = 0.05). CONCLUSIONS: Findings are consistent with the role of liver fat in diabetes pathogenesis. GGT may be a better diabetes predictor than ALT, but additional studies with directly determined liver fat content, ALT, and GGT are needed to confirm this finding. PMID- 19131467 TI - Growth factors and adipocytokines in prepubertal children born small for gestational age: relation to insulin resistance. AB - OBJECTIVE: The aim of this study was to test whether being born small for gestational age (SGA) has an impact on adiponectin and leptin levels and the IGF system in relation to insulin sensitivity, taking into consideration the severity of growth restriction. RESEARCH DESIGN AND METHODS: Serum levels of adiponectin, leptin, fasting glucose, fasting insulin (I(F)), the homeostasis model assessment insulin resistance index (HOMA-IR), IGF-1, free IGF-1, IGF-binding protein (IGFBP)-1 and -3, total cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides were evaluated in 57 children at age 4-10 years. Of these, 32 had been born appropriate size for gestational age (AGA) and 25 SGA (14 in the <3rd percentile and 11 in the 3rd-10th percentile). RESULTS; The SGA 3rd-10th percentile children were already insulin resistant at prepubertal age (I(F) 39.6 +/- 16.8 vs. 27 +/- 12 pmol/l, P < 0.01, and HOMA-IR 1.4 +/- 0.6 vs. 0.95 +/- 0.42 in SGA vs. AGA children, P < 0.05). Their IGF-1 and IGFBP-3 concentrations were significantly lower than those in AGA children (160.4 +/- 66.2 vs. 207 +/- 66.8 microg/l, P < 0.05 and 2.3 +/- 0.4 vs. 3.51 +/- 1.21 mg/l in SGA vs. AGA children, P < 0.01). The SGA <3rd percentile children had higher adiponectin (15.6 +/- 5.7 mg/l, P < 0.05) and IGFBP-1 levels (113.5 +/- 33.9 microg/l, P < 0.05) than AGA children (11.3 +/- 6.6 mg/l and 90.8 +/- 24.2 microg/l, respectively) and lower IGF-1 and IGFBP-3 concentrations (162.6 +/- 68.4 microg/l, P < 0.05 and 2.4 +/- 0.7 mg/l, P < 0.01). They also had significantly lower waist circumference (P < 0.05). Leptin levels did not differ among groups, but an inverse correlation with IGFBP-1 (r = -0.55, P < 0.01) was found in the pooled SGA group. CONCLUSIONS: Intrauterine growth restriction appears to affect the IGF axis at prepubertal age, and its severity plays a role in insulin sensitivity. PMID- 19131468 TI - Synergy between adiposity, insulin resistance, metabolic risk factors, and inflammation in adolescents. AB - OBJECTIVE: The purpose of this study was to investigate relationships between inflammatory markers and components of a metabolic syndrome cluster in adolescents. RESEARCH DESIGN AND METHODS: This was a cross-sectional analysis of an Australian childhood cohort (n = 1,377) aged 14 years. Cluster analysis defined a "high-risk" group similar to adults with metabolic syndrome. Relevant measures were anthropometry, fasting insulin, glucose, lipids, inflammatory markers, liver function, and blood pressure. RESULTS: Of the children, 29% fell into a high-risk metabolic cluster group compared with 2% by a pediatric metabolic syndrome definition. Relative to the "low-risk" cluster, they had higher BMI (95% CI 19.5-19.8 vs. 24.5-25.4), waist circumference (centimeters) (95% CI 71.0-71.8 vs. 83.4-85.8), insulin (units per liter) (95% CI 1.7-1.8 vs. 3.5-3.9), homeostasis model assessment (95% CI 1.7-1.8 vs. 3.5-3.9), systolic blood pressure (millimeters of mercury) (95% CI 110.8-112.1 vs. 116.7-118.9), and triglycerides (millimoles per liter) (95% CI 0.78-0.80 vs. 1.25-1.35) and lower HDL cholesterol (millimoles per liter) (95% CI 1.44-1.48 vs. 1.20-1.26). Inflammatory and liver function markers were higher in the high-risk group: C reactive protein (CRP) (P < 0.001), uric acid (P < 0.001), alanine aminotransferase (ALT) (P < 0.001), and gamma-glutamyl transferase (GGT) (P < 0.001). The highest CRP, GGT, and ALT levels were restricted to overweight children in the high-risk group. CONCLUSIONS: Cluster analysis revealed a strikingly high proportion of 14 year olds at risk of cardiovascular disease related metabolic disorders. Adiposity and the metabolic syndrome cluster are synergistic in the pathogenesis of inflammation. Systemic and liver inflammation in the high-risk cluster is likely to predict diabetes, cardiovascular disease, and nonalcoholic fatty liver disease. PMID- 19131469 TI - Effects of self-management support on structure, process, and outcomes among vulnerable patients with diabetes: a three-arm practical clinical trial. AB - OBJECTIVE: Despite the importance of self-management support (SMS), few studies have compared SMS interventions, involved diverse populations, or entailed implementation in safety net settings. We examined the effects of two SMS strategies across outcomes corresponding to the Chronic Care Model. RESEARCH DESIGN AND METHODS: A total of 339 outpatients with poorly controlled diabetes from county-run clinics were enrolled in a three-arm trial. Participants, more than half of whom spoke limited English, were uninsured, and/or had less than a high school education, were randomly assigned to usual care, interactive weekly automated telephone self-management support with nurse follow-up (ATSM), or monthly group medical visits with physician and health educator facilitation (GMV). We measured 1-year changes in structure (Patient Assessment of Chronic Illness Care [PACIC]), communication processes (Interpersonal Processes of Care [IPC]), and outcomes (behavioral, functional, and metabolic). RESULTS: Compared with the usual care group, the ATSM and GMV groups showed improvements in PACIC, with effect sizes of 0.48 and 0.50, respectively (P < 0.01). Only the ATSM group showed improvements in IPC (effect sizes 0.40 vs. usual care and 0.25 vs. GMV, P < 0.05). Both SMS arms showed improvements in self-management behavior versus the usual care arm (P < 0.05), with gains being greater for the ATSM group than for the GMV group (effect size 0.27, P = 0.02). The ATSM group had fewer bed days per month than the usual care group (-1.7 days, P = 0.05) and the GMV group (-2.3 days, P < 0.01) and less interference with daily activities than the usual care group (odds ratio 0.37, P = 0.02). We observed no differences in A1C change. CONCLUSIONS: Patient-centered SMS improves certain aspects of diabetes care and positively influences self-management behavior. ATSM seems to be a more effective communication vehicle than GMV in improving behavior and quality of life. PMID- 19131470 TI - Impaired mitochondrial function and insulin resistance of skeletal muscle in mitochondrial diabetes. AB - OBJECTIVE: Impaired muscular mitochondrial function is related to common insulin resistance in type 2 diabetes. Mitochondrial diseases frequently lead to diabetes, which is mostly attributed to defective beta-cell mitochondria and secretion. RESEARCH DESIGN AND METHODS: We assessed muscular mitochondrial function and lipid deposition in liver (hepatocellular lipids [HCLs]) and muscle (intramyocellular lipids [IMCLs]) using (31)P/(1)H magnetic resonance spectroscopy and insulin sensitivity and endogenous glucose production (EGP) using hyperinsulinemic-euglycemic clamps combined with isotopic tracer dilution in one female patient suffering from MELAS (myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) syndrome and in six control subjects. RESULTS: The MELAS patient showed impaired insulin sensitivity (4.3 vs. 8.6 +/- 0.5 mg x kg(-1) x min(-1)) and suppression of EGP (69 vs. 94 +/- 1%), and her baseline and insulin-stimulated ATP synthesis were reduced (7.3 and 8.9 vs. 10.6 +/- 1.0 and 12.8 +/- 1.3 micromol x l(-1) x min(-1)) compared with those of the control subjects. HCLs and IMCLs were comparable between the MELAS patient and control subjects. CONCLUSIONS: Impairment of muscle mitochondrial fitness promotes insulin resistance and could thereby contribute to the development of diabetes in some patients with the MELAS syndrome. PMID- 19131471 TI - Aging, exercise, and muscle protein metabolism. AB - Aging is accompanied by a progressive loss of skeletal muscle mass and strength, leading to the loss of functional capacity and an increased risk of developing chronic metabolic disease. The age-related loss of skeletal muscle mass is attributed to a disruption in the regulation of skeletal muscle protein turnover, resulting in an imbalance between muscle protein synthesis and degradation. As basal (fasting) muscle protein synthesis rates do not seem to differ substantially between the young and elderly, many research groups have started to focus on the muscle protein synthetic response to the main anabolic stimuli, i.e., food intake and physical activity. Recent studies suggest that the muscle protein synthetic response to food intake is blunted in the elderly. The latter is now believed to represent a key factor responsible for the age-related decline in skeletal muscle mass. Physical activity and/or exercise stimulate postexercise muscle protein accretion in both the young and elderly. However, the latter largely depends on the timed administration of amino acids and/or protein before, during, and/or after exercise. Prolonged resistance type exercise training represents an effective therapeutic strategy to augment skeletal muscle mass and improve functional performance in the elderly. The latter shows that the ability of the muscle protein synthetic machinery to respond to anabolic stimuli is preserved up to very old age. Research is warranted to elucidate the interaction between nutrition, exercise, and the skeletal muscle adaptive response. The latter is needed to define more effective strategies that will maximize the therapeutic benefits of lifestyle intervention in the elderly. PMID- 19131472 TI - Carnosine loading and washout in human skeletal muscles. AB - Carnosine (beta-alanyl-L-histidine) is present in high concentrations in human skeletal muscles. The oral ingestion of beta-alanine, the rate-limiting precursor in carnosine synthesis, has been shown to elevate the muscle carnosine content both in trained and untrained humans. Little human data exist about the dynamics of the muscle carnosine content, its metabolic regulation, and its dependence on muscle fiber type. The present study aimed to investigate in three skeletal muscle types the supplementation-induced amplitude of carnosine synthesis and its subsequent elimination on cessation of supplementation (washout). Fifteen untrained males participated in a placebo-controlled double-blind study. They were supplemented for 5-6 wk with either 4.8 g/day beta-alanine or placebo. Muscle carnosine was quantified in soleus, tibialis anterior, and medial head of the gastrocnemius by proton magnetic resonance spectroscopy (MRS), before and after supplementation and 3 and 9 wk into washout. The beta-alanine supplementation significantly increased the carnosine content in soleus by 39%, in tibialis by 27%, and in gastrocnemius by 23% and declined post-supplementation at a rate of 2-4%/wk. Average muscle carnosine remained increased compared with baseline at 3 wk of washout (only one-third of the supplementation-induced increase had disappeared) and returned to baseline values within 9 wk at group level. Following subdivision into high responders (+55%) and low responders (+15%), washout period was 15 and 6 wk, respectively. In the placebo group, carnosine remained relatively constant with variation coefficients of 9-15% over a 3-mo period. It can be concluded that carnosine is a stable compound in human skeletal muscle, confirming the absence of carnosinase in myocytes. The present study shows that washout periods for crossover designs in supplementation studies for muscle metabolites may sometimes require months rather than weeks. PMID- 19131473 TI - Mental fatigue impairs physical performance in humans. AB - Mental fatigue is a psychobiological state caused by prolonged periods of demanding cognitive activity. Although the impact of mental fatigue on cognitive and skilled performance is well known, its effect on physical performance has not been thoroughly investigated. In this randomized crossover study, 16 subjects cycled to exhaustion at 80% of their peak power output after 90 min of a demanding cognitive task (mental fatigue) or 90 min of watching emotionally neutral documentaries (control). After experimental treatment, a mood questionnaire revealed a state of mental fatigue (P = 0.005) that significantly reduced time to exhaustion (640 +/- 316 s) compared with the control condition (754 +/- 339 s) (P = 0.003). This negative effect was not mediated by cardiorespiratory and musculoenergetic factors as physiological responses to intense exercise remained largely unaffected. Self-reported success and intrinsic motivation related to the physical task were also unaffected by prior cognitive activity. However, mentally fatigued subjects rated perception of effort during exercise to be significantly higher compared with the control condition (P = 0.007). As ratings of perceived exertion increased similarly over time in both conditions (P < 0.001), mentally fatigued subjects reached their maximal level of perceived exertion and disengaged from the physical task earlier than in the control condition. In conclusion, our study provides experimental evidence that mental fatigue limits exercise tolerance in humans through higher perception of effort rather than cardiorespiratory and musculoenergetic mechanisms. Future research in this area should investigate the common neurocognitive resources shared by physical and mental activity. PMID- 19131474 TI - Onset responses of ventilation and cerebral blood flow to hypercapnia in humans: rest and exercise. AB - The respiratory and cerebrovascular reactivity to changes in arterial Pco(2) (Pa(CO(2))) is an important mechanism that maintains CO(2) or pH homeostasis in the brain. It remains unclear, however, how cerebrovascular CO(2) reactivity might influence the respiratory chemoreflex. The purpose of the present study was therefore to examine the interaction between onset responses of the respiratory chemoreflex and middle cerebral artery (MCA) mean blood velocity (V(mean)) to hypercapnia (5.0% CO(2)-40% O(2)-balance N(2)) at rest and during dynamic exercise ( approximately 1.0 l/min O(2) consumption). Each onset response was evaluated using a single-exponential regression model consisting of the response time latency [CO(2)-response delay (t(0))] and time constant (tau). At rest, t(0) and tau data indicated that the MCA V(mean) onset response was faster than the ventilatory (Ve) response (P < 0.001). In contrast, during exercise, t(0) of Ve and MCA V(mean) onset responses were decreased. In addition, despite the enhanced Pa(CO(2)) response to CO(2) administration (P = 0.014), tau of MCA V(mean) tended to increase during exercise (P = 0.054), whereas tau of Ve decreased (P = 0.015). These findings indicate that 1) at rest, faster washout of CO(2) via cerebral vasodilation results in a reduced activation of the central chemoreflex and subsequent reduced Ve onset response, and 2) during exercise, despite higher rates of increasing Pa(CO(2)), the lack of change in the onset response of cerebral blood flow and reduced washout of CO(2) may act to augment the Ve onset response. PMID- 19131476 TI - Impairment of the autonomic nervous function during decompression sickness in swine. AB - Dysautonomia has been observed in many cardiac diseases; however, its effect in decompression sickness (DCS) has not been well examined largely due to the difficulty in obtaining experimental data in human or animal subjects. In this study, we examine how DCS affects the autonomic nervous system's (ANS) dynamics in swine. Baseline and post-DCS electrocardiograms were obtained via telemetry recordings and compared. These data were analyzed using both the power spectrum method and our recently developed principal dynamic mode (PDM) analysis. PDM is able to separate the dynamic tones of the sympathetic and parasympathetic nervous systems. Both methods demonstrated a statistically significant decrease (>55%; P < 0.05) in the dynamics of both branches of the autonomic nervous system in the swine with DCS compared with the control condition. In cardiac diseases such as myocardial infarction, ANS imbalance is often associated with a significant increase in sympathetic tone, which may or may not be counterbalanced by parasympathetic nervous activity. However, the effect of DCS is such that both branches of the ANS are depressed >55% compared with the control condition, suggesting impairment, but not imbalance, of the ANS. PMID- 19131475 TI - Exercise training during diabetes attenuates cardiac ryanodine receptor dysregulation. AB - The present study was undertaken to assess the effects of exercise training (ExT) initiated after the onset of diabetes on cardiac ryanodine receptor expression and function. Type 1 diabetes was induced in male Sprague-Dawley rats using streptozotocin (STZ). Three weeks after STZ injection, diabetic rats were divided into two groups. One group underwent ExT for 4 wk while the other group remained sedentary. After 7 wk of sedentary diabetes, cardiac fractional shortening, rate of rise of left ventricular pressure, and myocyte contractile velocity were reduced by 14, 36, 44%, respectively. Spontaneous Ca(2+) spark frequency increased threefold, and evoked Ca(2+) release was dyssynchronous with diastolic Ca(2+) releases. Steady-state type 2 ryanodine receptor (RyR2) protein did not change, but its response to Ca(2+) was altered. RyR2 also exhibited 1.8- and 1.5 fold increases in phosphorylation at Ser(2808) and Ser(2814). PKA activity was reduced by 75%, but CaMKII activity was increased by 50%. Four weeks of ExT initiated 3 wk after the onset of diabetes blunted decreases in cardiac fractional shortening and rate of left ventricular pressure development, increased the responsiveness of the myocardium to isoproterenol stimulation, attenuated the increase in Ca(2+) spark frequency, and minimized dyssynchronous and diastolic Ca(2+) releases. ExT also normalized the responsiveness of RyR2 to Ca(2+) activation, attenuated increases in RyR2 phosphorylation at Ser(2808) and Ser(2814), and normalized CaMKII and PKA activities. These data are the first to show that ExT during diabetes normalizes RyR2 function and Ca(2+) release from the sarcoplasmic reticulum, providing insights into mechanisms by which ExT during diabetes improves cardiac function. PMID- 19131477 TI - Sensitization of capsaicin-sensitive lung vagal afferents by anandamide in rats: role of transient receptor potential vanilloid 1 receptors. AB - Anandamide (AEA), an arachidonic acid derivative produced during inflammatory conditions, is an endogenous agonist of both transient receptor potential vanilloid 1 (TRPV1) receptors and cannabinoid CB1 receptors. Sensitization of capsaicin-sensitive lung vagal afferent (CSLVA) fibers by chemical mediators is important in the pathogenesis of hyperreactive airway diseases. We investigated the effect of the intravenous infusion of AEA (2 mg x kg(-1) x ml(-1), 0.5 ml/min for 2 min) on the sensitivity of CSLVA fibers to chemical and mechanical stimulation in anesthetized rats. In artificially ventilated rats, AEA infusion only mildly elevated the baseline activity of CSLVA fibers. However, CSLVA fiber responses to right atrial injection of capsaicin, AEA, or adenosine and to lung inflation (tracheal pressure = 30 cmH(2)O) were all markedly potentiated during AEA infusion, which reverted 20 min after termination of the infusion. The potentiating effect on the sensitivity of CSLVA fibers to adenosine injection or lung inflation was completely blocked by pretreatment with capsazepine (a TRPV1 receptor antagonist) but was unaffected by pretreatment with AM281 (a CB1 receptor antagonist). In spontaneously breathing rats, right atrial injection of adenosine evoked an apneic response that is presumably mediated through CSLVA fibers. Similarly, the adenosine-evoked apneic response was potentiated during AEA infusion, and this potentiating effect was also completely prevented by pretreatment with capsazepine. These results suggest that AEA infusion at the dose tested produces a mild activation of TRPV1 receptors and this nonspecifically increases CSLVA fiber sensitivity to chemical and mechanical stimulation. PMID- 19131478 TI - Multiple pathological events in exercised dystrophic mdx mice are targeted by pentoxifylline: outcome of a large array of in vivo and ex vivo tests. AB - The phosphodiesterases inhibitor pentoxifylline gained attention for Duchenne muscular dystrophy therapy for its claimed anti-inflammatory, antioxidant, and antifibrotic action. A recent finding also showed that pentoxifylline counteracts the abnormal overactivity of a voltage-independent calcium channel in myofibers of dystrophic mdx mice. The possible link between workload, altered calcium homeostasis, and oxidative stress pushed toward a more detailed investigation. Thus a 4- to 8-wk treatment with pentoxifylline (50 mg x kg(-1) x day(-1) ip) was performed in mdx mice, undergoing or not a chronic exercise on treadmill. In vivo, the treatment partially increased forelimb strength and enhanced resistance to treadmill running in exercised animals. Ex vivo, pentoxifylline restored the mechanical threshold, an electrophysiological index of calcium homeostasis, and reduced resting cytosolic calcium in extensor digitorum longus muscle fibers. Mn quenching and patch-clamp technique confirmed that this effect was paralleled by a drug-induced reduction of membrane permeability to calcium. The treatment also significantly enhanced isometric tetanic tension in mdx diaphragm. The plasma levels of creatine kinase and reactive oxygen species were both significantly reduced in treated-exercised animals. Dihydroethidium staining, used as an indicator of reactive oxygen species production, showed that pentoxifylline significantly reduced the exercise-induced increase in fluorescence in the mdx tibialis anterior muscle. A significant decrease in connective tissue area and profibrotic cytokine transforming growth factor-beta(1) was solely found in tibialis anterior muscle. In both diaphragm and gastrocnemius muscle, a significant increase in neural cell adhesion molecule-positive area was instead observed. This data supports the interest toward pentoxifylline and allows insight in the level of cross talk between pathogenetic events in workloaded dystrophic muscle. PMID- 19131480 TI - Endurance exercise activates matrix metalloproteinases in human skeletal muscle. AB - In the present study, the effect of exercise training on the expression and activity of matrix metalloproteinases (MMPs) in the human skeletal muscle was investigated. Ten subjects exercised one leg for 45 min with restricted blood flow and then exercised the other leg at the same absolute workload with unrestricted blood flow. The exercises were conducted four times per week for 5 wk. Biopsies were taken from the vastus lateralis muscles of both legs at rest before the training period, after 10 days and 5 wk of training, and 2 h after the first exercise bout for analysis of MMP and tissue inhibitor of metalloproteinase 1 (TIMP-1) mRNA, enzyme activity, and protein expression. Levels of MMP-2, MMP 14, and TIMP-1 mRNA in muscle tissue increased after 10 days of training regardless of blood flow condition. MMP-2 mRNA level in laser-dissected myofibers and MMP-2 activity in whole muscle increased with training. The level of MMP-9 mRNA and activity increased after the first bout of exercise. Although MMP-9 mRNA levels appeared to be very low, the activity of MMP-9 after a single bout of exercise was similar to that of MMP-2 after 10 days of exercise. MMP-2 and MMP-9 protein was both present throughout the extracellular matrix of the muscle, both around fibers and capillaries, but MMP-2 was also present within the skeletal muscle fibers. These results show that MMPs are activated in skeletal muscle in nonpathological conditions such as voluntary exercise. The expression and time pattern indicate differences between the MMPs in regards of production sites as well as in the regulating mechanism. PMID- 19131479 TI - Spinal P2X receptor modulates muscle pressor reflex via glutamate. AB - Static contraction of skeletal muscle evokes reflex increases in blood pressure and heart rate. Previous studies showed that P2X receptors located at the dorsal horn of the spinal cord play a role in modulating the muscle pressor reflex. P2X stimulation can alter release of the excitatory amino acid, glutamate (Glu). In this report, we tested the hypothesis that stimulation of P2X receptors enhances the concentrations of Glu ([Glu]) in the dorsal horn, and that blocking P2X receptors attenuates contraction-induced Glu increases and the resultant reflex pressor response. Contraction was elicited by electrical stimulation of the L(7) and S(1) ventral roots of 14 cats. Glu samples were collected from microdialysis probes inserted in the L(7) level of the dorsal horn of the spinal cord, and dialysate [Glu] was determined using the HPLC method. First, microdialyzing alpha,beta-methylene ATP (0.4 mM) into the dorsal horn significantly increased [Glu]. In addition, contraction elevated [Glu] from baseline of 536 +/- 53 to 1,179 +/- 192 nM (P < 0.05 vs. baseline), and mean arterial pressure by 39 +/- 8 mmHg in the control experiment. Microdialyzing the P2X receptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (10 mM) into the dorsal horn attenuated the contraction induced-Glu increase (610 +/- 128 to 759 +/- 147 nM; P > 0.05) and pressor response (16 +/- 3 mmHg, P < 0.05 vs. control). Our findings demonstrate that P2X modulates the cardiovascular responses to static muscle contraction by affecting the release of Glu in the dorsal horn of the spinal cord. PMID- 19131481 TI - Nonselective NOS inhibition blunts the sweat response to exercise in a warm environment. AB - The role of nitric oxide synthase (NOS) inhibition in modulating human thermoregulatory control of sweating and cutaneous dilation was examined in 10 subjects (5 men and 5 women). Three intradermal microdialysis probes were placed in nonglabrous skin of the dorsum of the forearm. The control site was perfused with 0.9% saline, while the two remaining sites were perfused with a nonselective NOS inhibitor: 10 mM N(G)-nitro-L-arginine (L-NAME) or 10 mM N(G)-monomethyl-L arginine (L-NMMA). Local sweat rate (SR) and skin blood flow (laser-Doppler velocimetry) were monitored directly over the path of the intradermal microdialysis probe while arterial blood pressure was measured in the opposite arm noninvasively. Thermoregulatory responses were induced by cycle ergometer exercise (60% peak oxygen consumption) in a warm environment (30 degrees C). Esophageal temperature increased 1.5 +/- 0.2 degrees C during the 30 min of exercise. The cutaneous dilator response between 5 and 30 min of exercise in the heat was attenuated by both 10 mM L-NAME and 10 mM L-NMMA (P < 0.05). However, 10 mM L-NAME was more effective in blunting the rise in cutaneous vascular conductance during exercise than L-NMMA (P < 0.05). NOS inhibition also reduced the rise in local SR between 10 and 30 min of exercise (P < 0.05). In this case, 10 mM L-NMMA was more effective in limiting the increase in local SR than 10 mM L NAME (P < 0.05). We conclude that local production of nitric oxide in the skin or around the sweat gland augments local SR and cutaneous dilation during exercise in the heat. PMID- 19131482 TI - Different contribution of muscle and liver lipid metabolism to endurance capacity and obesity susceptibility of mice. AB - We investigated strain differences in whole body energy metabolism, peripheral lipid metabolism, and energy metabolism-related gene expression and protein levels in BALB/c, C57BL/6J, and A/J mice to evaluate the relationship between endurance capacity, susceptibility to diet-induced obesity, and differences in lipid metabolism in muscle and liver. A high-fat diet significantly increased body weight and fat weight in C57BL/6J mice, but not in BALB/c and A/J mice. The endurance capacity of BALB/c mice was 52% greater than that of C57BL/6J mice and 217% greater than that of A/J mice. The respiratory exchange ratio was lowest in BALB/c mice, higher in C57BL/6J mice, and highest in A/J mice, which inversely correlated with the endurance capacity and fatty acid beta-oxidation activity in the muscle. Plasma lactate levels measured immediately after exercise were lowest in BALB/c mice and highest in A/J mice, although there was no difference under resting conditions, suggesting that carbohydrate breakdown is suppressed by enhanced fat utilization during exercise in BALB/c mice. On the other hand, the body weight increase induced by high-fat feeding was related to a reduced whole body energy expenditure, higher respiratory quotient, and lower fatty acid beta oxidation activity in the liver. In addition, beta-oxidation activity in the muscle and liver roughly paralleled the mRNA and protein levels of lipid metabolism-related molecules, such as peroxisome proliferator-activated receptor and medium-chain acyl-CoA dehydrogenase, in each tissue. These findings indicate that genetically determined basal muscle and liver lipid metabolism and responsiveness to exercise influence physical performance and obesity susceptibility. PMID- 19131483 TI - Creation of a standardized geometry of the human nasal cavity. AB - A novel, standardized geometry of the human nasal cavity was created by aligning and processing 30 sets of computed tomography (CT) scans of nasal airways of healthy subjects. Digital three-dimensional (3-D) geometries of the 60 single human nasal cavities (30 right and 30 mirrored left cavities) were generated from the CT scans and measurements of physical parameters of each single nasal cavity were performed. A methodology was developed to scale, orient, and align the nasal geometries, after which 2-D digital coronal cross-sectional slices were generated. With the use of an innovative image processing algorithm, median cross sectional geometries were created to match median physical parameters while retaining the unique geometric features of the human nasal cavity. From these idealized 2-D images, an original 3-D standardized median human nasal cavity was created. This new standardized geometry was compared against the original geometries of all subjects as well as limited existing data from the literature. The new model has potential for use as a geometric standard in future experimental and numerical studies of deposition of inhaled aerosols, as well as for use as a reference during diagnosis of unhealthy patients. The specific procedure developed could also be applied to build standard nasal geometries for different identifiable groups within the larger population. PMID- 19131484 TI - Counterpoint: maximal oxygen uptake is not limited by a central nervous system governor. PMID- 19131487 TI - Hard evidence for a central governor is still lacking! PMID- 19131488 TI - Last word on point:counterpoint: maximal oxygen uptake is/is not limited by a central nervous system governor. PMID- 19131489 TI - Last word on point:counterpoint: maximal oxygen uptake is/is not limited by a central nervous system governor. PMID- 19131490 TI - Comment on point: counterpoint: sickle cell trait should/should not be considered asymptomatic and as a benign condition during physical activity. PMID- 19131491 TI - Interpreting the phosphocreatine time constant in aerobically exercising skeletal muscle. PMID- 19131493 TI - European journal of echocardiography. Preface. PMID- 19131494 TI - Aortic stenosis. AB - Aortic stenosis is a common finding at echocardiography. This review describes the assessment of the aortic valve, the left ventricle, and the rest of the heart. It discusses grading the stenosis including cases in which there are discrepancies between the different echocardiographic measurements. It discusses the indications for and interpretation of stress echocardiography and the role of pressure recovery and wave reflectance. It describes, in outline, the medical management of aortic stenosis and indications for surgery. The minimum dataset is: peak instantaneous velocity, mean gradient, effective orifice area, LV dimensions and systolic function, aortic diameter, mitral valve appearance and function, RV size and function and PA pressure. PMID- 19131495 TI - Echocardiography for transcatheter aortic valve implantation. AB - Transcatheter aortic valve implantation (TAVI) is a new technology that treats severe aortic stenosis. The technology has specific and mandatory technical requirements that must be met to allow safe and successful procedures. The echocardiographic method for diagnosis of aortic stenosis, anatomical case selection, procedural guidance and management of complications is discussed in this context. The role of the imaging specialist is defined within the TAVI team aiming to provide good outcomes for this high risk cohort of patients. PMID- 19131496 TI - Structure and anatomy of the aortic root. AB - AIMS: Understanding the anatomy of the aortic root is particularly relevant in the current era of evolving management strategies including percutaneous and transcatheter therapeutic techniques for valve or device implantations. METHODS AND RESULTS: This review describes the aortic root as a composite structure of several elements, not only the valvar leaflets. The valvar leaflets have a unique shape with deep closure lines buttressed by the nodule of Arantius. The scalloped configuration of the hingelines of the leaflets crosses the ventriculo-arterial junction, leaving interleaflet fibrous triangles between the sinuses that are anatomically aortic but haemodynamically ventricular. The fibrous triangle between the right and the non-coronary leaflets is the guide to the location of the atrioventricular conduction bundle. The coronary orifices are located close to the level of the sinutubular junction. Variations in leaflet structure and their arrangements result in valvar stenosis or regurgitation, or both. Often, diseases of the aortic root involve more than one structural element. CONCLUSION: The leaflets and their hingelines, aortic sinuses, interleaflets triangles sinutubular junction, and ventriculo-arterial junction and their structures adjoining the junctions should be taken into account when considering the aortic root. Owing to its central location, the aortic root is in close proximity to all the cardiac chambers, the atrial septum, ventricular septum and the atrioventricular conduction bundle. PMID- 19131497 TI - Echocardiography in the emergency assessment of acute aortic syndromes. AB - Acute aortic syndrome (AAS) is a collective term for several life-threatening acute aortic conditions: aortic dissection, intramural haematoma (IMH), penetrating atherosclerotic ulcer, and traumatic transection. Mortality from acute ascending aortic (type A) dissection increases rapidly immediately after presentation, reaching 1-2% per hour for the first 48 h. Early surgical intervention is recommended for type A aortic dissection and has been shown to improve outcome. Transthoracic echocardiography is an extremely valuable, often overlooked, clinical tool in diagnosing and assessing AAS in the emergency setting. Although diagnostic sensitivity is suboptimal, it is very useful in assessing potential high risk features or complications, such as pericardial effusion, and diagnosing potential differential conditions. A negative transthoracic echocardiography (TTE), however, does not exclude AAS. In patients with a high risk of type A dissection or IMH (identified clinically or by TTE), the safest and most rapid 'gold standard' investigation is transoesophageal echocardiography, ideally performed with the cardiac surgical team standing by. This is of particular importance in the haemodynamically unstable patient. Transoesophageal echocardiography, helical CT, and MRI have similar diagnostic accuracy and, when there is diagnostic uncertainty or no indication for immediate intervention, should be used according to clinical need, local availability, and expertise. PMID- 19131499 TI - G-protein-coupled receptors, cholesterol and palmitoylation: facts about fats. AB - G-protein-coupled receptors (GPCRs) are integral membrane proteins, hence it is not surprising that a number of their structural and functional features are modulated by both proteins and lipids. The impact of interacting proteins and lipids on the assembly and signalling of GPCRs has been extensively investigated over the last 20-30 years, and a further impetus has been given by the proposal that GPCRs and/or their immediate signalling partners (G proteins) can partition within plasma membrane domains, termed rafts and caveolae, enriched in glycosphingolipids and cholesterol. The high content of these specific lipids, in particular of cholesterol, in the vicinity of GPCR transmembranes can affect GPCR structure and/or function. In addition, most GPCRs are post-translationally modified with one or more palmitic acid(s), a 16-carbon saturated fatty acid, covalently bound to cysteine(s) localised in the carboxyl-terminal cytoplasmic tail. The insertion of palmitate into the cytoplasmic leaflet of the plasma membrane can create a fourth loop, thus profoundly affecting GPCR structure and hence the interactions with intracellular partner proteins. This review briefly highlights how lipids of the membrane and the receptor themselves can influence GPCR organisation and functioning. PMID- 19131498 TI - Detection of coronary artery disease with perfusion stress echocardiography using a novel ultrasound imaging agent: two Phase 3 international trials in comparison with radionuclide perfusion imaging. AB - AIMS: To determine if perfusion stress echocardiography (PSE) with Imagify (perflubutane polymer microspheres) is comparable to stress perfusion imaging using (99m)Tc single photon emission computed tomography (SPECT) for coronary artery disease (CAD) detection. PSE is a novel technique for evaluating myocardial perfusion. RAMP (real-time assessment of myocardial perfusion)-1 and 2 were international, Phase 3 trials that evaluated the ability of PSE with Imagify, to detect CAD. METHODS AND RESULTS: Chronic, stable, chest pain patients (n=662) underwent Imagify PSE and gated SPECT imaging at rest and during dipyridamole stress. Independent blinded cardiologists [three PSE readers per trial, and four SPECT readers (one for RAMP-1, three for RAMP-2)] interpreted images. CAD was defined by quantitative coronary angiography or 90-day outcome with clinical review. Accuracy, sensitivity, and specificity were evaluated using non-inferiority analysis (one-sided alpha=0.025) compared with SPECT. SPECT results for RAMP-1 and -2 were: accuracy (70%, 67%), sensitivity (78%, 61%), and specificity (64%, 76%). Accuracy of all six PSE readers was non-inferior to SPECT (66-71%, PT, 643C>T and -693G>C in TNFSF11 gene. Bone mineral density (BMD) and biochemical markers were measured. In the osteoporotic women, femoral neck BMD (BMD-fn) showed associations with BsmI(VDR) and Cdx2(VDR) (P=0.015 and 0.047 respectively), and lumbar spine BMD (BMD-ls) with K3N(OPG) and -290C>T(TNFSF11) (P=0.021 and 0.017). No association with BMD was found in the non-osteoporotic women. In the pre-menopausal women, the Cdx2(VDR) polymorphism was associated with BMD-fn and total hip BMD (P=0.011 and 0.011). In elderly men, FokI(VDR) was associated with BMD-fn and BMD-ls (P=0.040 and 0.036). Interestingly, the 290C>T(TNFSF11)-K3N(OPG) combination was associated with BMD-th (P=0.041) in the osteoporotic women. In the non-osteoporotic women, the combination K3N(OPG) Cdx2(VDR) was associated with BMD-ls, BMD-th and BMD-fn (P=0.032, 0.049 and 0.022), and the combination -290C>T(TNFSF11)-K3N(OPG) with BMD-fn (P=0.029). For the first time, the presence of gene-gene interactions between VDR, OPG and TNFSF11 genes was studied. Our results strongly suggest further confirmation of their combined influence on larger cohorts. PMID- 19131501 TI - Stanniocalcin 2 is positively and negatively controlled by 1,25(OH)(2)D(3) and PTH in renal proximal tubular cells. AB - We have previously identified a second mammalian stanniocalcin (STC2) in humans and demonstrated that STC2 inhibits phosphate uptake in an opossum renal proximal tubular cell line (opossum kidney (OK) cells). However, the regulation of Stc2 gene expression in OK cells is not well understood. In this study, we identified the opossum Stc2 cDNA sequence. The opossum STC2 amino acid sequence had 78.8% homology with human STC2, and has a conserved putative N-linked glycosylation site. Next, we investigated the regulation of Stc2 gene expression by the classical calcium and phosphate-regulating factors 1,25(OH)(2)D(3) and PTH in OK cells. In western blot analysis using affinity-purified anti-STC2 antibody, the secretion of STC2 protein was stimulated by 1,25(OH)(2)D(3) in a dose-dependent manner. By contrast, PTH suppressed the induction of STC2 protein secretion by 1,25(OH)(2)D(3). Real-time PCR analysis revealed that Stc2 mRNA expression was increased by 1,25(OH)(2)D(3) in a dose- and time-dependent manner. In addition, actinomycin D, an RNA synthesis inhibitor, prevented the effects of 1,25(OH)(2)D(3) on Stc2 gene expression. On the other hand, PTH and phorbol 12,13 myristic acetate, a specific PKC activator, but not 8-bromo-cyclic AMP, a specific PKA activator, reduced the mRNA levels of Stc2. In addition, Go6976, a specific PKC inhibitor, abolished the downregulation of Stc2 mRNA expression by PTH. Furthermore, we demonstrated that the renal Stc2 mRNA expression was increased by 1,25(OH)(2)D(3) and decreased by PTH in vivo. These results suggest that STC2 is positively and negatively controlled by 1,25(OH)(2)D(3) and PTH in renal proximal tubular cells. PMID- 19131502 TI - Screening for membrane hormone receptor expression in primary aldosteronism. AB - OBJECTIVE: The investigation of expression and functional relevance of G-protein coupled receptors in primary aldosteronism (PA) by molecular and clinical studies. PATIENTS AND METHODS: Tissues of 14 aldosterone-producing adenomas (APA), of one unilateral adrenal hyperplasia and of six healthy adult adrenal glands; 12 patients with confirmed PA due to APA; (n=5), idiopathic hyperaldosteronism (n=7) and 8 control subjects (C). The tissues were subjected to a quantitative PCR for determination of mRNA expression levels of AT2R1, GIPR, MC2R, GnRHR, LHR, TRHR, TSHR, glucagon-R, V1aR, V2R, and 5-HT4R. The patients and controls were enrolled in a test protocol consisting of stimulation by posture, mixed meal, ACTH, GnRH, TRH, glucagon, vasopressin, and metoclopramide (MCP). Three patients could be analyzed by both studies. A positive response was defined as an aldosterone increase of more than 50% following stimulation. RESULTS: All the tissues revealed AT2R1, MC2R, AVPR, and 5-HT4R mRNA expression. LHR mRNA was found in normal adrenals and 13 adrenal tumors. By contrast with normal adrenals tumorous adrenal tissue expressed GnRHR mRNA (4/15) and TSHR mRNA (1/15). Both the patients and controls responded to posture, ACTH, glucagon, AVP, and MCP. Specific responses were seen in one patient following TRH and three patients following GnRH stimulation. CONCLUSIONS: We provide evidence for peptide hormone responsiveness to various peptide hormones in patients with PA, including GnRH and TRH. A good correlation between clinical and molecular testing could be observed, making an involvement of the receptor expressed in PA possible. PMID- 19131504 TI - Quality of life is reduced in patients with Klinefelter syndrome on androgen replacement therapy. AB - CONTEXT: Klinefelter syndrome (KS) is a genetic disorder, characterized by an XXY karyotype, hypergonadotrophic hypogonadism and infertility, which affects approximately 1 in 700 men. KS has also been associated with neuropsychological alterations. OBJECTIVE: To investigate whether quality of life (QoL) is reduced in patients with KS on androgen treatment. DESIGN: Observational study. METHODS: Questionnaires (RAND-36) were sent to 46 KS patients on androgen treatment who regularly visit the outpatient clinic of an academic medical centre. QoL scores were compared with a Dutch male reference group. RESULTS: Forty (87%) questionnaires were returned which were suitable for evaluation. KS patients had significantly lower (worse) scores on eight of the nine domains of the RAND questionnaire compared with the male reference group. In KS patients, higher education was associated with significantly better QoL scores. CONCLUSIONS: QoL is reduced in patients with KS on androgen treatment. PMID- 19131503 TI - Small tumor size as favorable prognostic factor after adrenalectomy in Conn's adenoma. AB - OBJECTIVE: Primary aldosteronism (PA) due to aldosterone-producing adenoma (APA) is the most common curable form of secondary hypertension. DESIGN: In order to evaluate blood pressure outcome after adrenalectomy for APA and to identify new favorable prognostic factors, data from 42 consecutive APA patients who underwent adrenalectomy were collected from 2005 to 2007. METHODS: Renin-angiotensin aldosterone system (upright and postsaline infusion test), serum and urinary electrolytes, office and ambulatory blood pressure monitoring were evaluated at baseline and after a follow-up of 2.7+/-2.2 years. Drug history and adenoma size at morphological evaluation were also collected. RESULTS: Multiple regression analysis showed that, before surgery, patients with a small adenoma (diameter <20 mm) displayed higher postsaline aldosterone values (P=0.0001), and lower serum potassium levels (P=0.020), than patients with adenoma >20 mm. Before surgery, mineralocorticoid receptor (MR) antagonists were used in patients with small APA in greater percentage than patients with bigger adenomas (64 vs 30% respectively, P=0.037). At follow-up, blood pressure normalized in 63% of the subjects. Recovered patients had a shorter duration of hypertension (P=0.038), and a smaller adenoma size (P=0.035). Receiver operating characteristic curves showed that a duration of hypertension 85%) in the first 114 amino acids, there is considerable sequence divergence at their carboxy ends. The CGbeta-subunit terminates with a unique carboxyl-terminal extension (115-145; carboxyl-terminal peptide), which contains four O-linked oligosaccharides, whereas the LHbeta subunit bears a hydrophobic heptapeptide (115-121) at its carboxy terminus. LH is released through the regulated pathway in the pituitary, whereas CG is secreted constitutively from the placenta. We previously demonstrated in rat somatotroph derived GH(3) cells that the LH is associated primarily with a regulated routing, and although the majority of CG was released constitutively from the cells, there was a fraction that was segregated through the regulated pathway. Moreover, we showed that the LHbeta heptapeptide is a determinant for the regulated secretion of LH. Given that the primary evolutionary change between LHbeta and CGbeta occurred at the carboxy terminus, these data suggested that the presence of the CGbeta carboxyl-terminal peptide region is responsible for the constitutive secretion of CG. A CG114 mutant (CGDeltaT) was constructed and expressed in GH(3) cells. Steady-state labeling and pulse-chase experiments demonstrated that the CGDeltaT entered the regulated pathway resulting in over 4-fold increase in the intracellular pool. The secretagogue, forskolin, stimulated CGDeltaT release over 3-fold, which was accompanied by a parallel intracellular decrease, and only marginal stimulation of CG was seen. Immunofluorescence demonstrated a unique membrane pattern of staining for CGDeltaT compared with dispersed cytoplasmic puncta for CG. Stimulation with forskolin caused a significant reduction in the relative fluorescence of CGDeltaT cells compared with a minor reduction for CG. These data show that the CGDeltaT analog resembles LH in its intracellular trafficking, further supporting the hypothesis that determinants at the carboxyl terminal end of the CGbeta-subunit evolved from the LHbeta-subunit primarily to overcome the slow release and intracellular storage of LH resulting in rapid secretion of CG from the placenta. PMID- 19131509 TI - Distinct dysregulation of lipid metabolism by unliganded thyroid hormone receptor isoforms. AB - Thyroid hormone receptors (TRs) play critical roles in energy homeostasis. To understand the role of TRs in lipid homeostasis in vivo, we adopted the loss-of function approach by creating knock-in mutant mice with targeted mutation in the TRalpha gene (TRalpha1PV mouse) or TRbeta gene (TRbetaPV mouse). The PV mutation, identified in a patient with resistance to thyroid hormone, exhibits potent dominant-negative activity. Here we show that in contrast to TRalpha1PV mouse, TRbetaPV mice exhibited no significant reduction in WAT but had significant increases in serum free fatty acids and total triglycerides. Moreover, the liver of TRbetaPV mice was markedly increased (33%) with excess lipid accumulation, but the liver mass of TRalpha1PV mouse was decreased (23%) with paucity of lipids. These results indicate that apo-TRbeta and apo-TRalpha1 exerted distinct abnormalities in lipid metabolism. Further biochemical analyses indicate that increased lipogenic enzyme expression, activated peroxisome proliferator activated receptor gamma (Ppargamma) signaling, and decreased fatty acid beta oxidation activity contributed to the adipogenic steatosis and lipid accumulation in the liver of TRbetaPV mice. In contrast, the expression of lipogenic enzymes and Ppargamma was decreased in the liver of TRalpha1PV mice. These results suggest that the regulation of genes critical for lipid metabolism by TRs in the liver is isoform dependent. These results indicate that apo-TRbeta and apo TRalpha1 had different effects on lipid metabolism and that both TR isoforms contribute to the pathogenesis of lipid metabolism in hypothyroidism. PMID- 19131510 TI - Involvement of G protein-coupled receptor 30 (GPR30) in rapid action of estrogen in primate LHRH neurons. AB - Previously, we have reported that 17beta-estradiol (E(2)) induces an increase in firing activity of primate LH-releasing hormone (LHRH) neurons. The present study investigates whether E(2) alters LHRH release as well as the pattern of intracellular calcium ([Ca(2+)](i)) oscillations and whether G protein-coupled receptor 30 (GPR30) plays a role in mediating the rapid E(2) action in primate LHRH neurons. Results are summarized: 1) E(2), the nuclear membrane-impermeable estrogen, estrogen-dendrimer conjugate, and the plasma membrane-impermeable estrogen, E(2)-BSA conjugate, all stimulated LHRH release within 10 min of exposure; 2) whereas the estrogen receptor antagonist, ICI 182,780, did not block the E(2)-induced LHRH release, E(2) application to cells treated with pertussis toxin failed to induce LHRH release; 3) GPR30 mRNA was expressed in olfactory placode cultures, and GPR30 protein was expressed in a subset of LHRH neurons; 4) pertussis toxin treatment blocked the E(2)-induced increase in [Ca(2+)](i) oscillations; 5) knockdown of GPR30 in primate LHRH neurons by transfection with small interfering RNA (siRNA) for GPR30 completely abrogated the E(2)-induced changes in [Ca(2+)](i) oscillations, whereas transfection with control siRNA did not; 6) the estrogen-dendrimer conjugate-induced increase in [Ca(2+)](i) oscillations also did not occur in LHRH neurons transfected with GPR30 siRNA; and 7) G1, a GPR30 agonist, resulted in changes in [Ca(2+)](i) oscillations, similar to those observed with E(2). Collectively, E(2) induces a rapid excitatory effect on primate LHRH neurons, and this rapid action of E(2) appears to be mediated, in part, through GPR30. PMID- 19131511 TI - Inhibition of androgen receptor transcriptional activity as a novel mechanism of action of arsenic. AB - Environmental sodium arsenite is a toxin that is associated with male infertility due to decreased and abnormal sperm production. Arsenic trioxide (ATO), another inorganic trivalent semimetal, is an effective therapy for acute promyelocytic leukemia, and there is investigation of its possible efficacy in prostate cancer. However, the mechanism of arsenic action in male urogenital tract tissues is not clear. Because the androgen receptor (AR) plays an important role in spermatogenesis and prostate cancer, we explored the possibility that trivalent arsenic regulates AR function. We found that arsenic inhibited AR transcriptional activity in prostate cancer and Sertoli cells using reporter gene assays testing several androgen response element-containing regions and by assessing native target gene expression. Arsenic inhibition of AR activity was not due to down regulation of AR protein levels, decreased hormone binding to AR, disruption of AR nuclear translocation, or interference with AR-DNA binding in vitro. However, chromatin immunoprecipitation studies revealed that arsenic inhibited AR recruitment to an AR target gene enhancer in vivo. Consistent with a deficiency in AR-chromatin binding, arsenic disrupted AR amino and carboxyl termini interaction. Furthermore, ATO caused a significant decrease in prostate cancer cell proliferation that was more pronounced in cells expressing AR compared with cells depleted of AR. In addition, inhibition of AR activity by ATO and by the AR antagonist, bicalutamide, was additive. Thus, arsenic-induced male infertility may be due to inhibition of AR activity. Further, because AR is an important target in prostate cancer therapy, arsenic may serve as an effective therapeutic option. PMID- 19131512 TI - Connective tissue growth factor (CTGF) inactivation leads to defects in islet cell lineage allocation and beta-cell proliferation during embryogenesis. AB - The factors necessary for normal pancreatic islet morphogenesis have not been well characterized. Here we report that connective tissue growth factor (CTGF) is involved in the establishment of normal islet endocrine cell ratio and architecture. CTGF is a secreted protein known to modulate several growth factor signaling pathways including TGF-beta, BMP, and Wnt. Although its role in pancreatic diseases such as pancreatitis and pancreatic cancer are well documented, a role for CTGF in normal pancreas development and function has heretofore not been examined. Using a lacZ-tagged CTGF allele, we describe for the first time the expression pattern of CTGF in the developing pancreas and the requirement of CTGF for normal islet morphogenesis and embryonic beta-cell proliferation. CTGF is highly expressed in pancreatic ductal epithelium and vascular endothelium, as well as at lower levels in developing insulin(+) cells, but becomes down-regulated in beta-cells soon after birth. Pancreata from CTGF null embryos have an increase in glucagon(+) cells with a concomitant decrease in insulin(+) cells, and show defects in islet morphogenesis. Loss of CTGF also results in a dramatic decrease in beta-cell proliferation at late gestation. Unlike CTGF null embryos, CTGF heterozygotes survive past birth and exhibit a range of islet phenotypes, including an intermingling of islet cell types, increased number of glucagon(+) cells, and beta-cell hypertrophy. PMID- 19131513 TI - Localized and efficient curli nucleation requires the chaperone-like amyloid assembly protein CsgF. AB - Elucidation of the early events in amyloidogenesis is key to understanding the pathology of, and developing therapies for, amyloid diseases. Critical informants about these early events are amyloid assembly proteins that facilitate the transition from monomer to amyloid fiber. Curli are a functional amyloid whose in vivo polymerization requires a dedicated nucleator protein, CsgB, and an assembly protein, CsgF. Here we demonstrate that without CsgF, curli subunits are released from the cell into the media and are inefficiently polymerized, resulting in fewer and mislocalized curli fibers. CsgF is secreted to the cell surface, where it mediates the cell-association and protease-resistance of the CsgB nucleator, suggesting that CsgF is required for specific localization and/or chaperoning of CsgB for full nucleator activity. CsgF is thus critical to achieve localized and efficient nucleation of fiber subunits into functional, cell-associated amyloid. PMID- 19131514 TI - Allelic recombination between distinct genomic locations generates copy number diversity in human beta-defensins. AB - Beta-defensins are small secreted antimicrobial and signaling peptides involved in the innate immune response of vertebrates. In humans, a cluster of at least 7 of these genes shows extensive copy number variation, with a diploid copy number commonly ranging between 2 and 7. Using a genetic mapping approach, we show that this cluster is at not 1 but 2 distinct genomic loci approximately 5 Mb apart on chromosome band 8p23.1, contradicting the most recent genome assembly. We also demonstrate that the predominant mechanism of change in beta-defensin copy number is simple allelic recombination occurring in the interval between the 2 distinct genomic loci for these genes. In 416 meiotic transmissions, we observe 3 events creating a haplotype copy number not found in the parent, equivalent to a germ line rate of copy number change of approximately 0.7% per gamete. This places it among the fastest-changing copy number variants currently known. PMID- 19131515 TI - Location of KCNE1 relative to KCNQ1 in the I(KS) potassium channel by disulfide cross-linking of substituted cysteines. AB - The cardiac-delayed rectifier K(+) current (I(KS)) is carried by a complex of KCNQ1 (Q1) subunits, containing the voltage-sensor domains and the pore, and auxiliary KCNE1 (E1) subunits, required for the characteristic I(KS) voltage dependence and kinetics. To locate the transmembrane helix of E1 (E1-TM) relative to the Q1 TM helices (S1-S6), we mutated, one at a time, the first four residues flanking the extracellular ends of S1-S6 and E1-TM to Cys, coexpressed all combinations of Q1 and E1 Cys-substituted mutants in CHO cells, and determined the extents of spontaneous disulfide-bond formation. Cys-flanking E1-TM readily formed disulfides with Cys-flanking S1 and S6, much less so with the S3-S4 linker, and not at all with S2 or S5. These results imply that the extracellular flank of the E1-TM is located between S1 and S6 on different subunits of Q1. The salient functional effects of selected cross-links were as follows. A disulfide from E1 K41C to S1 I145C strongly slowed deactivation, and one from E1 L42C to S6 V324C eliminated deactivation. Given that E1-TM is between S1 and S6 and that K41C and L42C are likely to point approximately oppositely, these two cross-links are likely to favor similar axial rotations of E1-TM. In the opposite orientation, a disulfide from E1 K41C to S6 V324C slightly slowed activation, and one from E1 L42C to S1 I145C slightly speeded deactivation. Thus, the first E1 orientation strongly favors the open state, while the approximately opposite orientation favors the closed state. PMID- 19131516 TI - Active site dynamics in the zinc-dependent medium chain alcohol dehydrogenase superfamily. AB - Despite being the subject of intensive investigations, many aspects of the mechanism of the zinc-dependent medium chain alcohol dehydrogenase (MDR) superfamily remain contentious. We have determined the high-resolution structures of a series of binary and ternary complexes of glucose dehydrogenase, an MDR enzyme from Haloferax mediterranei. In stark contrast to the textbook MDR mechanism in which the zinc ion is proposed to remain stationary and attached to a common set of protein ligands, analysis of these structures reveals that in each complex, there are dramatic differences in the nature of the zinc ligation. These changes arise as a direct consequence of linked movements of the zinc ion, a zinc-bound bound water molecule, and the substrate during progression through the reaction. These results provide evidence for the molecular basis of proton traffic during catalysis, a structural explanation for pentacoordinate zinc ion intermediates, a unifying view for the observed patterns of metal ligation in the MDR family, and highlight the importance of dynamic fluctuations at the metal center in changing the electrostatic potential in the active site, thereby influencing the proton traffic and hydride transfer events. PMID- 19131517 TI - Local conformational dynamics in alpha-helices measured by fast triplet transfer. AB - Coupling fast triplet-triplet energy transfer (TTET) between xanthone and naphthylalanine to the helix-coil equilibrium in alanine-based peptides allowed the observation of local equilibrium fluctuations in alpha-helices on the nanoseconds to microseconds time scale. The experiments revealed faster helix unfolding in the terminal regions compared with the central parts of the helix with time constants varying from 250 ns to 1.4 micros at 5 degrees C. Local helix formation occurs with a time constant of approximately 400 ns, independent of the position in the helix. Comparing the experimental data with simulations using a kinetic Ising model showed that the experimentally observed dynamics can be explained by a 1-dimensional boundary diffusion with position-independent elementary time constants of approximately 50 ns for the addition and of approximately 65 ns for the removal of an alpha-helical segment. The elementary time constant for helix growth agrees well with previously measured time constants for formation of short loops in unfolded polypeptide chains, suggesting that helix elongation is mainly limited by a conformational search. PMID- 19131518 TI - A distinct H2A.X isoform is enriched in Xenopus laevis eggs and early embryos and is phosphorylated in the absence of a checkpoint. AB - Histone H2A.X is an H2A variant present in multicellular organisms that is specifically phosphorylated on the serine in the C-terminal consensus sequence, canonically "SQEY," in response to DNA damage. We have recently shown the significance of phosphorylation of the penultimate tyrosine for maintenance and processing of the DNA damage response in mammalian cells. Here, we report the identification of distinct H2A.X variants in the eggs and early embryos of the frog Xenopus laevis that contain a C-terminal SQEF, among other changes; we have denoted these proteins as "H2A.X-F." H2A.X-F is present only in late-staged oocytes, eggs, and premidblastula transition embryos and is not present in somatic cells. Similar unannotated isoforms were identified in other rapidly developing aquatic species, such as Xenopus tropicalis, goldfish, and zebrafish, and in Arabidopsis and chickpea. Furthermore, we demonstrate by mass spectrometry and phospho-specific antibodies that H2A.X-F is phosphorylated in the absence of exogenous DNA damage, in both actively dividing, unperturbed embryos and cell free egg extract in the absence and presence of DNA damage and S-phase checkpoint conditions. We propose that this isoform may be involved in modulating the cellular response to the rapid early cell cycles in externally developing species. PMID- 19131519 TI - Sulfation of o-demethyl apixaban: enzyme identification and species comparison. AB - Apixaban, a potent and highly selective factor Xa inhibitor, is currently under development for treatment of arterial and venous thrombotic diseases. The O demethyl apixaban sulfate is a major circulating metabolite in humans but circulates at lower concentrations relative to parent in animals. The aim of this study was to identify the sulfotransferases (SULTs) responsible for the sulfation reaction. Apixaban undergoes O-demethylation catalyzed by cytochrome P450 enzymes to O-demethyl apixaban, and then is conjugated by SULTs to form O-demethyl apixaban sulfate. Of the five human cDNA-expressed SULTs tested, SULT1A1 and SULT1A2 exhibited significant levels of catalytic activity for formation of O demethyl apixaban sulfate, and SULT1A3, SULT1E1, and SULT2A1 showed much lower catalytic activities. In human liver S9, quercetin, a highly selective inhibitor of SULT1A1 and SULT1E1, inhibited O-demethyl apixaban sulfate formation by 99%; 2,6-dichloro-4-nitrophenol, another inhibitor of SULT1A1, also inhibited this reaction by >90%; estrone, a competitive inhibitor for SULT1E1, had no effect on this reaction. The comparable K(m) values for formation of O-demethyl apixaban sulfate were 41.4 microM (human liver S9), 36.8 microM (SULT1A1), and 70.8 microM (SULT1A2). Because of the high level of expression of SULT1A1 in liver and its higher level of catalytic activity for formation of O-demethyl apixaban sulfate, SULT1A1 might play a major role in humans for formation of O-demethyl apixaban sulfate. O-Demethyl apixaban was also investigated in liver S9 of mice, rats, rabbits, dogs, monkeys, and humans. The results indicated that liver S9 samples from dogs, monkeys, and humans had higher activities for formation of O-demethyl apixaban sulfate than those of mice, rats, and rabbits. PMID- 19131520 TI - Measurement of membrane-bound human heme oxygenase-1 activity using a chemically defined assay system. AB - Heme oxygenase (HO) catalyzes heme degradation in a reaction requiring NADPH cytochrome P450 reductase (CPR). Although most studies with HO used a soluble 30 kDa form, lacking the C-terminal membrane-binding region, recent reports show that the catalytic behavior of this enzyme is very different if this domain is retained; the overall activity was elevated 5-fold, and the K(m) for CPR decreased approximately 50-fold. The goal of these studies was to accurately measure HO activity using a coupled assay containing purified biliverdin reductase (BVR). This allows measurement of bilirubin formation after incorporation of full-length CPR and heme oxygenase-1 (HO-1) into a membrane environment. When rat liver cytosol was used as the source of partially purified BVR, the reaction remained linear for 2 to 3 min; however, the reaction was only linear for 10 to 30 s when an equivalent amount of purified, human BVR (hBVR) was used. This lack of linearity was not observed with soluble HO-1. Optimal formation of bilirubin was achieved with concentrations of bovine serum albumin (0.25 mg/ml) and hBVR (0.025-0.05 microM), but neither supplement increased the time that the reaction remained linear. Various concentrations of superoxide dismutase had no effect on the reaction; however, when catalase was included, the reactions were linear for at least 4 to 5 min, even at high CPR levels. These results not only show that HO-1-generated hydrogen peroxide leads to a decrease in HO-1 activity but also provide for a chemically defined system to be used to examine the function of full-length HO-1 in a membrane environment. PMID- 19131521 TI - Mechanism of gender-divergent UDP-glucuronosyltransferase mRNA expression in mouse liver and kidney. AB - UDP-glucuronosyltransferases (UGTs) catalyze the addition of glucuronic acid to endo- and xenobiotics, increasing hydrophilicity and enhancing elimination. Gender-divergent glucuronidation rates are observed in humans and rats, and gender differences in UGT mRNA levels have been observed in rodents. The purpose of this study was to establish the hormonal regulation of gender-dependent Ugt mRNA expression in mouse liver and kidney. Therefore, three mouse models were used to characterize the involvement of sex hormones and gender-specific growth hormone (GH) secretion patterns, including 1) hypophysectomized mice treated with male- or female-pattern GH, testosterone, or 17beta-estradiol; 2) GH releasing hormone receptor-deficient little (lit/lit) mice treated with male- or female pattern GH; and 3) gonadectomized mice treated with testosterone or 17beta estradiol. Messenger RNA expression of mouse Ugt isozymes was determined by the branched DNA assay. In C57BL/6 mice, male-predominant expression of Ugt2b1 and Ugt2b38 was observed in liver and kidney, respectively. Female-predominant expression was observed for Ugt1a1 and Ugt1a5 in liver and Ugt1a2 in kidney. In liver, regulation of Ugt1a1 and Ugt1a5 expression was attributed to repression of Ugt mRNA by male-pattern GH secretion. Conversely, regulation of Ugt2b1 expression in liver was attributed to male-pattern GH secretion. In kidney, regulation of Ugt2b38 expression was attributed to inductive effects by testosterone. Conversely, Ugt1a2 expression in kidney was negatively regulated by testosterone. In conclusion, gender differences in mouse Ugt mRNA expression were influenced by male-pattern GH secretion in liver, whereas gender differences were regulated by the effects of androgens in kidney. PMID- 19131522 TI - Absorption, metabolism, and excretion of darunavir, a new protease inhibitor, administered alone and with low-dose ritonavir in healthy subjects. AB - Absorption, metabolism, and excretion of darunavir, an inhibitor of human immunodeficiency virus protease, was studied in eight healthy male subjects after a single oral dose of 400 mg of [(14)C]darunavir given alone (unboosted subjects) or with ritonavir [100 mg b.i.d. 2 days before and 7 days after darunavir administration (boosted subjects)]. Plasma exposure to darunavir was 11-fold higher in boosted subjects. Total recoveries of radioactivity in urine and feces were 93.9 and 93.5% of administered radioactivity in unboosted and boosted subjects, respectively. The most radioactivity was recovered in feces (81.7% in unboosted subjects and 79.5% in boosted subjects, compared with 12.2 and 13.9% recovered in urine, respectively). Darunavir was extensively metabolized in unboosted subjects, mainly by carbamate hydrolysis, isobutyl aliphatic hydroxylation, and aniline aromatic hydroxylation and to a lesser extent by benzylic aromatic hydroxylation and glucuronidation. Total excretion of unchanged darunavir accounted for 8.0% of the dose in unboosted subjects. Boosting with ritonavir resulted in significant inhibition of carbamate hydrolysis, isobutyl aliphatic hydroxylation, and aniline aromatic hydroxylation but had no effect on aromatic hydroxylation at the benzylic moiety, whereas excretion of glucuronide metabolites was markedly increased but still represented a minor pathway. Total excretion of unchanged darunavir accounted for 48.8% of the administered dose in boosted subjects as a result of the inhibition of darunavir metabolism by ritonavir. Unchanged darunavir in urine accounted for 1.2% of the administered dose in unboosted subjects and 7.7% in boosted subjects, indicating a low renal clearance. Darunavir administered alone or with ritonavir was well tolerated. PMID- 19131523 TI - Physiologically based pharmacokinetics of matrine in the rat after oral administration of pure chemical and ACAPHA. AB - ACAPHA, a botanical drug for the treatment of human esophageal cancer in China, is under investigation as a lung cancer chemoprevention agent at the BC Cancer Agency (Vancouver, BC, Canada). Little or no information is available on the pharmacokinetics of ACAPHA in animals. The objectives of this study were as follows: to examine the disposition kinetics of matrine, a bioactive marker of ACAPHA in the rat; to develop a physiologically based pharmacokinetic (PBPK) model for pure matrine; and to characterize the absorption and clearance of crude matrine in ACAPHA-treated rats using the PBPK model. Pure matrine (15 mg/kg) or crude matrine in the form of ACAPHA (0.38 or 3.8 g/kg) was administered to the rat by gavages. The rats were sacrificed at different time points postdosing. Blood and major organs were removed from the rat, extracted with toluene/butanol, and quantified for matrine using gas chromatography-mass spectrometry. An 11 compartment, flow-limited PBPK model of matrine was developed. The PBPK model was able to simulate closely the empirical data of rats treated with pure matrine. Because the absorption and clearance of crude matrine in ACAPHA-treated rats could not be parameterized a priori, they were estimated by fitting the experimental data to the PBPK model. Results of the study show that pure matrine is absorbed and eliminated by the rat at faster rates than crude matrine. Moreover, the ACAPHA matrix may change the pharmacokinetics of matrine in the rat significantly. The PBPK model is a valuable tool to gain insights into the disposition kinetics of a botanical drug. PMID- 19131524 TI - Silencing the breast cancer resistance protein expression and function in caco-2 cells using lentiviral vector-based short hairpin RNA. AB - A series of stable breast cancer resistance protein (BCRP, ABCG2) knockdown cell lines were produced by transduction of Caco-2 cells with lentiviral vector-based short hairpin RNA (shRNA). Caco-2 cell is a human intestinal-derived cell line widely used to study intestinal drug absorption. Caco-2 expresses three apical drug efflux transporters: BCRP, P-glycoprotein (P-gp; ABCB1), and multidrug resistance protein 2 (MRP2, ABCC2). BCRP and P-gp in particular play a significant role in pharmacokinetics because of their expression at several key interfaces. Overexpression of BCRP in cancer cells may also be a mechanism of tumor resistance to chemotherapeutic drugs. The goal of this study was to engineer and characterize Caco-2 cell clones with stable knockdown of BCRP expression. The shRNA/BCRP lentiviral particles were used to infect a stable clone of Caco-2 cells. Expression of BCRP was monitored using quantitative polymerase chain reaction (qPCR), Western blotting, immunofluorescence microscopy, and bidirectional transport of probe substrates, estrone-3-sulfate (E3S), and pheophorbide A (PhA). Based on qPCR, expression of BCRP mRNA was knocked down in five clones with a maximum of 97% silencing in clone D. Silencing of BCRP gene expression was maintained for at least 25 passages. Expression of BCRP protein was also reduced significantly. Functionally, BCRP knockdown was reflected in significant reduction of the efflux ratio of E3S and PhA. Clone D in particular should be a useful model for identifying and characterizing P-gp substrates and inhibitors without interference from BCRP and/or MRP2. In addition, it can be used in conjunction with wild-type or vector control Caco-2 cells to identify BCRP substrates. PMID- 19131526 TI - General overview of lung transplantation and review of organ allocation. AB - Lung transplantation is an established treatment option for patients with a wide variety of end-stage lung diseases. For patients with end-stage lung disease, lung transplant can prolong life substantially; however, the survival statistics for lung transplants still pale compared with other solid organ transplants. Acute cellular rejection (ACR) is common after lung transplantation occurring in up to 90 percent of patients. Chronic allograft rejection, manifest as bronchiolitis obliterans syndrome (BOS), remains the "Achilles heel" to the long term success of lung transplantation. Unfortunately, BOS is common after lung transplantation, occurring in a majority of patients by 5 years after transplant. Candidates for lung transplantation should have near-end-stage lung disease with a limited life expectancy. Allocation of organs today is based upon need and survivability of the operation, and there is a high likelihood of improvement in quality of life. Details of the advances in this fascinating field are included in the several articles in this issue. PMID- 19131525 TI - Strategic plan for pediatric respiratory diseases research: an NHLBI working group report. AB - The Division of Lung Diseases of the National Heart, Lung, and Blood Institute (NHLBI) recently held a workshop to identify gaps in our understanding and treatment of childhood lung diseases and to define strategies to enhance translational research in this field. Leading experts with diverse experience in both laboratory and patient-oriented research reviewed selected areas of pediatric lung diseases, including perinatal programming and epigenetic influences; mechanisms of lung injury, repair, and regeneration; pulmonary vascular disease; sleep and control of breathing; and the application of novel translational methods to enhance personalized medicine. This report summarizes the proceedings of this workshop and provides recommendations for emphasis on targeted areas for future investigation. The priority areas identified for research in pediatric pulmonary diseases included: (1) epigenetic and environmental influences on lung development that program pediatric lung diseases; (2) injury, regeneration, and repair in the developing lung; (3) pulmonary vascular disease in children; (4) development and adaptation of ventilatory responses to postnatal life; (5) nonatopic wheezing: aberrant large airway development or injury?; (6) strategies to improve assessment, diagnosis, and treatment of pediatric respiratory diseases; and (7) predictive and personalized medicine for children. PMID- 19131527 TI - Selection of candidates for lung transplantation. AB - The selection of candidates for lung transplantation requires an appreciation of the natural history of lung disease to determine when the disease has entered an advanced and imminently life-threatening stage. It also requires an understanding of the impact of preexisting medical comorbidities on transplant outcomes. Finally, selection is influenced by the particular metric by which outcomes are judged to be successful (e.g., maximizing short-term net survival benefit versus maximizing post-transplant longevity). This article will discuss general and disease-specific criteria used to select patients for lung transplantation and determine the appropriate timing of listing. It will highlight current guidelines put forth by the International Society of Heart and Lung Transplantation and will explore the published data upon which these guidelines are based. PMID- 19131528 TI - Lung donor selection and management. AB - Lung transplantation is still limited by the shortage of suitable donor organs. This results in long waiting times for listed patients with a substantial risk (10-15%) of dying before transplantation. All efforts to increase donor awareness through legislation, public campaigns, and training of transplant coordinators and medical ICU staff should be encouraged. Only a minority of cadaveric donors meets the preset ideal lung donor criteria, leaving many transplantable lungs untouched. Donor lung utilization can be further improved by careful selection of extended criteria donors, by active participation of transplant teams in donor management, and by verifying as often as possible the quality of lungs in the donor hospital by a member of the transplant team. This article aims to update the current evidence from the literature to identify and select potential lung donors and to manage cadaveric donors to maximally increase the organ yield for lung transplantation. PMID- 19131529 TI - Primary graft dysfunction. AB - Primary graft dysfunction (PGD) is a severe form of ischemia/reperfusion acute lung injury that is a major cause of early morbidity and mortality after lung transplantation. Survivors of PGD have worse long-term lung function and an increased chance of developing bronchiolitis obliterans syndrome (BOS), the manifestation of chronic rejection. This review examines the current state of PGD research in the context of a recent consensus statement by the International Society for Heart and Lung Transplantation (ISHLT) designed to standardize the definition of PGD in clinical research efforts. This article will review this definition and issues surrounding it, outcome studies examining the long-term effects of PGD, and the established clinical risk factors for PGD. Translational studies exploring the pathogenesis of PGD will be highlighted, and the current state of PGD prevention and management will also be described. Finally, we will summarize efforts at finding genetic and molecular markers for PGD and discuss future directions for PGD research. PMID- 19131530 TI - Immunosuppression for lung transplantation. AB - Immunosuppression remains the mainstay of therapy for successful outcomes after lung transplantation. The need for optimal immunosuppression became evident to maintain long-term graft survival and to navigate the delicate balance between infection and rejection. Over the past two decades, immunosuppression for solid organ transplantation has evolved to target multiple immune pathways with the hope of decreasing both acute and chronic allograft rejection. Although current maintenance therapy after lung transplantation typically includes a calcineurin inhibitor, antimetabolite and corticosteroid therapy, newer therapies including induction therapy with biological agents, mTOR inhibitors, and salvage therapies including photopheresis and total lymphoid irradiation have emerged as alternate therapeutic options. This review will discuss both the current immunosuppressive medications that are used as well as different therapeutic combinations that are currently employed. In addition, we will discuss the current literature regarding the efficacy of these agents in lung transplantation. PMID- 19131532 TI - Surgical techniques: lung transplant and lung volume reduction. AB - Patients with end-stage emphysema can be offered lung volume reduction surgery (LVRS), lung transplantation, or unilateral lung transplantation combined with contralateral LVRS if necessary, depending on multiple factors including age, lung function parameters, lobar predominance, and whether the disease is uni- or bilateral. Lung transplant is a complex and well-established therapeutic modality for patients with end-stage lung disease. The ideal candidate for LVRS is a patient with severe upper-lobe predominant emphysema and markedly impaired exercise capacity. Other groups may benefit from the procedure, but results are likely to be less good. The objective of this chapter is to describe the surgical techniques of bilateral lung transplantation and lung volume reduction surgery as performed at the University of Toronto. PMID- 19131531 TI - Acute rejection and humoral sensitization in lung transplant recipients. AB - Despite the recent introduction of many improved immunosuppressive agents for use in transplantation, acute rejection affects up to 55% of lung transplant recipients within the first year after transplant. Acute lung allograft rejection is defined as perivascular or peribronchiolar mononuclear inflammation. Although histopathologic signs of rejection often resolve with treatment, the frequency and severity of acute rejections represent the most important risk factor for the subsequent development of bronchiolitis obliterans syndrome (BOS), a condition of progressive airflow obstruction that limits survival to only 50% at 5 years after lung transplantation. Recent evidence demonstrates that peribronchiolar mononuclear inflammation (also known as lymphocytic bronchiolitis) or even a single episode of minimal perivascular inflammation significantly increase the risk for BOS. We comprehensively review the clinical presentation, diagnosis, histopathologic features, and mechanisms of acute cellular lung rejection. In addition, we consider emerging evidence that humoral rejection occurs in lung transplantation, characterized by local complement activation or the presence of antibody to donor human leukocyte antigens (HLA). We discuss in detail methods for HLA antibody detection as well as the clinical relevance, the mechanisms, and the pathologic hallmarks of humoral injury. Treatment options for cellular rejection include high-dose methylprednisolone, antithymocyte globulin, or alemtuzumab. Treatment options for humoral rejection include intravenous immunoglobulin, plasmapheresis, or rituximab. A greater mechanistic understanding of cellular and humoral forms of rejection and their role in the pathogenesis of BOS is critical in developing therapies that extend long-term survival after lung transplantation. PMID- 19131533 TI - Airway complications and management after lung transplantation: ischemia, dehiscence, and stenosis. AB - Overall survival rates of lung transplantation have improved since the first human lung transplantation was performed. A decline in the incidence of airway complications (AC) had been a key feature to achieve the current outcomes. Several proposed risk factors to the development of airway complications have been identified, ranging from the surgical technique to the immunosuppressive regimen. There are essentially six different airway complications post-lung transplantation. The most frequently reported complication is bronchial stenosis. Other complications include bronchial dehiscence, exophytic excessive granulation tissue formation, tracheo-bronchomalacia, bronchial fistulas, and endobronchial infections. The management of post-transplant bronchial complications needs a multispecialty team approach. Prevention of some complications may be possible by early and aggressive medical management as well as by using certain surgical techniques for transplantation. Interventional bronchoscopic procedures, including balloon bronchoplasty, cryotherapy, laser photoresection, electrocautery, high-dose endobronchial brachytherapy, and bronchial stents are among the armamentarium. Also, medical management, like antibiotic prophylaxis and therapy for endobronchial infections, or noninvasive positive-pressure ventilation in case of bronchomalacia, are used to treat an AC. In some cases, different surgical approaches are occasionally required. In this article we review the risk factors, the clinical presentation, the diagnostic methods, as well as the management options of the most common AC after lung transplantation. PMID- 19131534 TI - Infections relevant to lung transplantation. AB - Allograft infection after lung transplantation has a significant impact on the outcome and can be a diagnostic challenge. The increased susceptibility of the pulmonary allograft to infection is due to its direct contact with environmental microbes by inhalation, concurrent immunosuppression, and the impaired clearance mechanisms after denervation of the transplanted lung. The possible spectrum of microorganisms infecting the allograft after lung transplantation is broad, but commonly includes Pseudomonas aeruginosa, cytomegalovirus, community-acquired respiratory viruses, and Aspergillus species. Prophylactic antimicrobial treatment regimens after surgery reduced the incidence of infections. However, preventive strategies for reducing infectious complications used by different transplant centers are still heterogeneous, and many questions regarding efficacy remain unanswered. PMID- 19131535 TI - Medical complications of lung transplantation. AB - As short- and long-term survival rates for lung transplantation continue to improve, and as more lung transplantations are occurring with each year, a multitude of medical complications are encountered by the clinician. This article reviews the long-term non-pulmonary noninfectious medical complications that arise beyond the postoperative period in patients who have undergone lung transplantation. This article reviews the development of renal failure, diabetes, cardiovascular complications of hypertension and atherosclerosis, osteoporosis and avascular necrosis, hematologic complications, thromboembolic disease, gastrointestinial complications, neurologic complications, and malignancy, including post-transplant lymphoproliferative disorder. PMID- 19131536 TI - Chronic lung allograft rejection: mechanisms and therapy. AB - Lung transplantation is a therapeutic option for patients with end-stage pulmonary disorders. Unfortunately, due to post-lung transplant complications, both infectious and noninfectious, it is only a treatment and not a cure. Importantly, despite induction combined with triple or quadruple maintenance immunosuppressive therapy, chronic lung rejection, in the form of obliterative bronchiolitis or its clinical correlate bronchiolitis obliterans syndrome (BOS), continues to be highly prevalent and is the major limitation to long-term survival. In this review we evaluate the presentation, diagnosis, histopathology, pathologic mechanisms, risk factors, and prevention/treatment options for BOS. A better understanding of the risk factors and how it relates to the pathologic mechanisms of chronic lung allograft rejection should lead to better pharmacologic targets to prevent/treat this syndrome without increasing the recipient's risk for infections. PMID- 19131537 TI - Pediatric lung transplantation. AB - Pediatric lung and heart-lung transplantation are viable therapeutic interventions for end stage pulmonary parenchymal or pulmonary vascular diseases. Issues specific to pediatrics include unique diagnoses and increased need for mechanical ventilation before transplant and increased complications related to infection both before and after transplant. Although outcomes for children as a group are similar to those for adults, young children often fare better, perhaps in part due to reduced incidence of acute and chronic rejection. As in other solid organs, long-term outcomes in adolescents are poor. Increased focus on improving adherence in this age group will be important. Similar to adults, bronchiolitis obliterans remains the major late complication. Uniform treatment protocols and multicenter pediatric studies will be required to ultimately overcome this pervasive problem and improve pediatric lung transplant outcomes. PMID- 19131538 TI - Technology and outcomes assessment in lung transplantation. AB - Lung transplantation offers the hope of prolonged survival and significant improvement in quality of life to patients that have advanced lung diseases. However, the medical literature lacks strong positive evidence and shows conflicting information regarding survival and quality of life outcomes related to lung transplantation. Decisions about the use of lung transplantation require an assessment of trade-offs: do the potential health and quality of life benefits outweigh the potential risks and harms? No amount of theoretical reasoning can resolve this question; empiric data are needed. Rational analyses of these trade offs require valid measurements of the benefits and harms to the patients in all relevant domains that affect survival and quality of life. Lung transplant systems and registries mainly focus outcomes assessment on patient survival on the waiting list and after transplantation. Improved analytic approaches allow comparisons of the survival effects of lung transplantation versus continued waiting. Lung transplant entities do not routinely collect quality of life data. However, the medical community and the public want to know how lung transplantation affects quality of life. Given the huge stakes for the patients, the providers, and the healthcare systems, key stakeholders need to further support quality of life assessment in patients with advanced lung disease that enter into the lung transplant systems. Studies of lung transplantation and its related technologies should assess patients with tools that integrate both survival and quality of life information. Higher quality information obtained will lead to improved knowledge and more informed decision making. PMID- 19131539 TI - Nonmedical therapy for chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a category of diseases with chronic airflow obstruction and hyperinflation. The GOLD committee and the American Thoracic Society/European Respiratory Society have published detailed, evidence-based reviews of management, providing stepped-care algorithms for pharmacologic and nonpharmacologic therapy. Over the past several decades, this has led to numerous nonpharmacologic approaches to ameliorate symptoms in these patients. PMID- 19131540 TI - Patient perceptions of a community-based care coordination system. AB - Community health workers (CHWs) work with health professionals to improve health outcomes by facilitating community-based health education and increase access and continuity to health services within a community. Uninsured, low-income participants of a community-based program, Project Access Dallas, participated in focus group sessions for determining participants' perceptions of CHW effectiveness and participants' abilities to independently manage their health needs. Of the 95 adults invited, 24 (25.3%) attended. Participants reported that CHWs are an invaluable asset in learning how to navigate the health care system, obtaining appointments and being better able to care for themselves with CHW emotional/psychological support. Results suggest that CHWs in a case management model improved patient comprehension of health issues, patient navigation through a health care system, and patients' abilities to independently manage health issues. Implementation of CHWs within a case management model appears to be an effective mechanism for providing health services to underserved populations. PMID- 19131541 TI - Process evaluation and participatory methods in an obesity-prevention media campaign for Mexican Americans. AB - To address obesity and related morbidities, community-based participatory research (CBPR) strategies were employed to design and evaluate a Spanish language media campaign promoting physical activity and healthful food choices among Mexican Americans. Process evaluation including content analyses on types and focus of media messages was conducted. Focus groups assessed appeal and trustworthiness of messages. All media campaign products featured role models and experts. Campaign messages primarily (91%) appeared in TV morning show segments. Newsletters presented individual and family role model stories. A majority of newsletters (68%) were distributed through churches and "promotora" outreach efforts. CBPR lends itself to the selection and tailoring of evidence-based media campaigns. Moreover, CBPR guidance resulted in media messages that were credible and appealing to audience. Process evaluation strategies that gather information from the community provide solid evidence for how to modify the campaign to best meet audience expectations. PMID- 19131542 TI - Grape seed extract inhibits VEGF expression via reducing HIF-1alpha protein expression. AB - Grape seed extract (GSE) is a widely consumed dietary supplement that has antitumor activity. Here, we have investigated the inhibitory effect of GSE on the expression of vascular endothelial growth factor (VEGF) and the mechanism underlying this action. We found that GSE inhibited VEGF messenger RNA (mRNA) and protein expression in U251 human glioma cells and MDA-MB-231 human breast cancer cells. GSE inhibited transcriptional activation of the VEGF gene through reducing protein but not mRNA expression of hypoxia-inducible factor (HIF) 1alpha. The inhibitory effect of GSE on HIF-1alpha expression was mainly through inhibiting HIF-1alpha protein synthesis rather than promoting protein degradation. Consistent with this result, GSE-suppressed phosphorylation of several important components involved in HIF-1alpha protein synthesis, such as Akt, S6 kinase and S6 protein. Furthermore, in the MDA-MB-231 tumor, we found that GSE treatment inhibited the expression of VEGF and HIF-1alpha and the phosphorylation of S6 kinase without altering the subcellular localization of HIF-1alpha, correlating with reduced vessel density and tumor size. Depletion of polyphenol with polyvinylpyrrolidone abolished the inhibitory activity of GSE, suggesting a water soluble fraction of polyphenol in GSE is responsible for the inhibitory activity. Taken together, our results indicate that GSE inhibits VEGF expression by reducing HIF-1alpha protein synthesis through blocking Akt activation. This finding provides new insight into the mechanisms of anticancer activity of GSE and reveals a novel molecular mechanism underlying the antiangiogenic action of GSE. PMID- 19131543 TI - Nicotine stimulates pancreatic cancer xenografts by systemic increase in stress neurotransmitters and suppression of the inhibitory neurotransmitter gamma aminobutyric acid. AB - Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer mortality in Western countries. We have shown previously that four representative human PDAC cell lines were regulated by beta-adrenoreceptors via cyclic adenosine 3',5' monophosphate (cAMP)-dependent signaling. In the current study, we have tested the hypothesis that nicotine stimulates the growth of PDAC xenografts in nude mice by increasing the systemic levels of the stress neurotransmitters adrenaline and noradrenaline, which are the physiological agonists for beta-adrenoreceptors and that inhibition by gamma-aminobutyric acid (GABA) of the adenylyl cyclase dependent pathway downstream of adrenoreceptors blocks this effect. The size of xenografts from PDAC cell line Panc-1 was determined 30 days after inoculation of the cancer cells. Stress neurotransmitters in serum as well as cAMP in the cellular fraction of blood and in tumor tissue were assessed by immunoassays. Levels of GABA, its synthesizing enzymes GAD65 and GAD67 and beta-adrenergic signaling proteins in the tumor tissue were determined by western blotting. Nicotine significantly increased the systemic levels of adrenaline, noradrenaline and cAMP while increasing xenograft size and protein levels of cAMP, cyclic AMP response element-binding protein and p-extracellular signal-regulated kinase 1/2 in the tumor tissue. Nicotine additionally reduced the protein levels of both GAD isozymes and GABA in tumor tissue. Treatment with GABA abolished these responses to nicotine and blocked the development of xenografts in mice not exposed to nicotine. These findings suggest that the development and progression of PDAC is subject to significant modulation by stimulatory stress neurotransmitters and inhibitory GABA and that treatment with GABA may be useful for marker-guided cancer intervention of PDAC. PMID- 19131544 TI - PDLIM2 suppresses human T-cell leukemia virus type I Tax-mediated tumorigenesis by targeting Tax into the nuclear matrix for proteasomal degradation. AB - The mechanisms by which the human T-cell leukemia virus type I (HTLV-I) Tax oncoprotein deregulates cellular signaling for oncogenesis have been extensively studied, but how Tax itself is regulated remains largely unknown. Here we report that Tax was negatively regulated by PDLIM2, which promoted Tax K48-linked polyubiquitination. In addition, PDLIM2 recruited Tax from its functional sites into the nuclear matrix where the polyubiquitinated Tax was degraded by the proteasome. Consistently, PDLIM2 suppressed Tax-mediated signaling activation, cell transformation, and oncogenesis both in vitro and in animal. Notably, PDLIM2 expression was down-regulated in HTLV-I-transformed T cells, and PDLIM2 reconstitution reversed the tumorigenicity of the malignant cells. These studies indicate that the counterbalance between HTLV-I/Tax and PDLIM2 may determine the outcome of HTLV-I infection. These studies also suggest a potential therapeutic strategy for cancers and other diseases associated with HTLV-I infection and/or PDLIM2 deregulation. PMID- 19131546 TI - Clinical characterization of acute myeloid leukemia with myelodysplasia-related changes as defined by the 2008 WHO classification system. AB - Although some studies have validated the 2001 World Health Organization (WHO) classification of acute myeloid leukemia (AML), including the importance of multilineage dysplasia, others have suggested that multilineage dysplasia correlates with unfavorable cytogenetics but has no independent impact on prognosis. In 2008, the revised WHO classification has expanded this category into "AML with myelodysplasia-related changes" (AML-MRC). We evaluated the clinical, pathologic, cytogenetic, and molecular features of 100 AML patients using the 2008 WHO criteria. Patients underwent genetic screening for NPM1, FLT3 ITD, FLT3-D835, and CEBPA mutations. Compared with patients with AML, not otherwise specified, patients with AML-MRC were significantly older (P= .014), presented with a lower hemoglobin (P= .044), more frequently expressed CD14 (P= .048), and exhibited a decreased frequency of CEBPA mutations (P= .001). Multivariate analysis indicated that patients with AML-MRC had a significantly worse overall survival, progression-free survival, and complete response compared with AML-not otherwise specified (all P< .001). These data support the clinical, morphologic, and cytogenetic criteria for this 2008 WHO AML category. PMID- 19131545 TI - Acute mixed lineage leukemia in children: the experience of St Jude Children's Research Hospital. AB - To characterize the biology and optimal therapy of acute mixed-lineage leukemia in children, we reviewed the pathologic and clinical features, including response to therapy, of 35 patients with mixed-lineage leukemia. The majority of cases (91%) had blasts cells that simultaneously expressed either T-lineage plus myeloid markers (T/myeloid, n = 20) or B-lineage plus myeloid markers (B/myeloid, n = 12). Overall survival rates for the B/myeloid and T/myeloid subgroups were not significantly different from each other or from the rate for acute myeloid leukemia (AML) but were inferior to the outcome in children with acute lymphoblastic leukemia (ALL). Patients who failed to achieve complete remission with AML-directed therapy could often be induced with a regimen of prednisone, vincristine, and L-asparaginase. Analysis of gene-expression patterns identified a subset of biphenotypic leukemias that did not cluster with T-cell ALL, B progenitor ALL, or AML. We propose that treatment for biphenotypic leukemia begin with one course of AML-type induction therapy, with a provision for a switch to lymphoid-type induction therapy with a glucocorticoid, vincristine, and L asparaginase if the patient responds poorly. We also suggest that hematopoietic stem cell transplantation is often not required for cure of these patients. PMID- 19131547 TI - The role of cytogenetic abnormalities as a prognostic marker in primary myelofibrosis: applicability at the time of diagnosis and later during disease course. AB - Although cytogenetic abnormalities are important prognostic factors in myeloid malignancies, they are not included in current prognostic scores for primary myelofibrosis (PMF). To determine their relevance in PMF, we retrospectively examined the impact of cytogenetic abnormalities and karyotypic evolution on the outcome of 256 patients. Baseline cytogenetic status impacted significantly on survival: patients with favorable abnormalities (sole deletions in 13q or 20q, or trisomy 9 +/- one other abnormality) had survivals similar to those with normal diploid karyotypes (median, 63 and 46 months, respectively), whereas patients with unfavorable abnormalities (rearrangement of chromosome 5 or 7, or > or = 3 abnormalities) had a poor median survival of 15 months. Patients with abnormalities of chromosome 17 had a median survival of only 5 months. A model containing karyotypic abnormalities, hemoglobin, platelet count, and performance status effectively risk-stratified patients at initial evaluation. Among 73 patients assessable for clonal evolution during stable chronic phase, those who developed unfavorable or chromosome 17 abnormalities had median survivals of 18 and 9 months, respectively, suggesting the potential role of cytogenetics as a risk factor applicable at any time in the disease course. Dynamic prognostic significance of cytogenetic abnormalities in PMF should be further prospectively evaluated. PMID- 19131548 TI - Plasma from stored packed red blood cells and MHC class I antibodies causes acute lung injury in a 2-event in vivo rat model. AB - Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion death. We hypothesize that TRALI requires 2 events: (1) the clinical condition of the patient and (2) the infusion of antibodies against MHC class I antigens or the plasma from stored blood. A 2-event rat model was developed with saline (NS) or endotoxin (LPS) as the first event and the infusion of plasma from packed red blood cells (PRBCs) or antibodies (OX18 and OX27) against MHC class I antigens as the second event. ALI was determined by Evans blue dye leak from the plasma to the bronchoalveolar lavage fluid (BALF), protein and CINC-1 concentrations in the BALF, and the lung histology. NS-treated rats did not evidence ALI with any second events, and LPS did not cause ALI. LPS-treated animals demonstrated ALI in response to plasma from stored PRBCs, both prestorage leukoreduced and unmodified, and to OX18 and OX27, all in a concentration-dependent fashion. ALI was neutrophil (PMN) dependent, and OX18/OX27 localized to the PMN surface in vivo and primed the oxidase of rat PMNs. We conclude that TRALI is the result of 2 events with the second events consisting of the plasma from stored blood and antibodies that prime PMNs. PMID- 19131549 TI - Haptoglobin preserves the CD163 hemoglobin scavenger pathway by shielding hemoglobin from peroxidative modification. AB - Detoxification and clearance of extracellular hemoglobin (Hb) have been attributed to its removal by the CD163 scavenger receptor pathway. However, even low-level hydrogen peroxide (H(2)O(2)) exposure irreversibly modifies Hb and severely impairs Hb endocytosis by CD163. We show here that when Hb is bound to the high-affinity Hb scavenger protein haptoglobin (Hp), the complex protects Hb from structural modification by preventing alpha-globin cross-links and oxidations of amino acids in critical regions of the beta-globin chain (eg, Trp15, Cys93, and Cys112). As a result of this structural stabilization, H(2)O(2) exposed Hb-Hp binds to CD163 with the same affinity as nonoxidized complex. Endocytosis and lysosomal translocation of oxidized Hb-Hp by CD163-expressing cells were found to be as efficient as with nonoxidized complex. Hp complex formation did not alter Hb's ability to consume added H(2)O(2) by redox cycling, suggesting that within the complex the oxidative radical burden is shifted to Hp. We provide structural and functional evidence that Hp protects Hb when oxidatively challenged with H(2)O(2) preserving CD163-mediated Hb clearance under oxidative stress conditions. In addition, our data provide in vivo evidence that unbound Hb is oxidatively modified within extravascular compartments consistent with our in vitro findings. PMID- 19131550 TI - Aberrant splicing of folylpolyglutamate synthetase as a novel mechanism of antifolate resistance in leukemia. AB - Folylpoly-gamma-gluatamate synthetase (FPGS) catalyzes the polyglutamylation and thus intracellular retention of folates and antifolates (eg, methotrexate; MTX) through the addition of multiple glutamate equivalents to their gamma-carboxyl residue. Since polyglutamylation of antifolates is crucial for their pharmacological activity in leukemia, loss of FPGS function results in decreased cellular levels of polyglutamylation-dependent antifolates and consequent drug resistance. Whereas resistance to pulse exposure to antifolates is frequently associated with loss of FPGS activity, the underlying molecular mechanism remains elusive. Here we explored the molecular basis of antifolate resistance in human MTX-resistant leukemia cell lines displaying marked loss of FPGS activity. We demonstrate that these MTX-resistant cells exhibit impaired splicing of FPGS mRNA based on intron retention and/or exon skipping, thereby resulting in loss of FPGS function due to premature translation termination. Furthermore, analysis of FPGS transcripts in blood or bone marrow specimens from patients with acute lymphoblastic leukemia revealed exon 12 skipping, both at diagnosis and at relapse, the latter of which occurs after high-dose MTX-containing chemotherapy. These results constitute the first demonstration of the loss of FPGS function via aberrant mRNA splicing, thereby resulting in loss of antifolate retention and drug resistance. The clinical ramifications of these novel findings are discussed. PMID- 19131551 TI - Human erythrocytes bind and inactivate type 5 adenovirus by presenting Coxsackie virus-adenovirus receptor and complement receptor 1. AB - Type 5 adenovirus (Ad5) is a human pathogen that has been widely developed for therapeutic uses, with only limited success to date. We report here the novel finding that human erythrocytes present Coxsackie virus-adenovirus receptor (CAR) providing an Ad5 sequestration mechanism that protects against systemic infection. Interestingly, erythrocytes from neither mice nor rhesus macaques present CAR. Excess Ad5 fiber protein or anti-CAR antibody inhibits the binding of Ad5 to human erythrocytes and cryo-electron microscopy shows attachment via the fiber protein of Ad5, leading to close juxtaposition with the erythrocyte membrane. Human, but not murine, erythrocytes also present complement receptor (CR1), which binds Ad5 in the presence of antibodies and complement. Transplantation of human erythrocytes into nonobese diabetic/severe combined immunodeficiency mice extends blood circulation of intravenous Ad5 but decreases its extravasation into human xenograft tumors. Ad5 also shows extended circulation in transgenic mice presenting CAR on their erythrocytes, although it clears rapidly in transgenic mice presenting erythrocyte CR1. Hepatic infection is inhibited in both transgenic models. Erythrocytes may therefore restrict Ad5 infection (natural and therapeutic) in humans, independent of antibody status, presenting a formidable challenge to Ad5 therapeutics. "Stealthing" of Ad5 using hydrophilic polymers may enable circumvention of these natural virus traps. PMID- 19131552 TI - Dose-dense induction with sequential high-dose cytarabine and mitoxantone (S-HAM) and pegfilgrastim results in a high efficacy and a short duration of critical neutropenia in de novo acute myeloid leukemia: a pilot study of the AMLCG. AB - Dose density during early induction has been demonstrated to be one of the prime determinants for treatment efficacy in acute myeloid leukemia (AML). The German AML Cooperative Group has therefore piloted a dose-dense induction regimen sequential high-dose AraC and mitoxantrone followed by pegfilgrastim (S-HAM) in which 2 induction cycles are applied over 11 to 12 days instead of 25 to 29 days as used in conventional double induction, thereby increasing dose density 2-fold. Of 172 de novo AML patients (excluding acute promyelocytic leukemia), 61% reached a complete remission, 22% a complete remission with incomplete peripheral recovery, 7% had persistent leukemia, 10% died (early death) resulting in an overall response rate of 83%. Kaplan-Meier estimated survival at 2 years was 61% for the whole group (patients with unfavorable karyotypes, 38%; patients with favorable karyotypes, 69%; patients with intermediate karyotypes, 75%) after S HAM treatment. Importantly, the compression of the 2 induction cycles into the first 11 to 12 days of treatment was beneficial for normal hematopoiesis as demonstrated by a significantly shortened duration of critical neutropenia of 31 days compared with 46 days after conventionally timed double induction. PMID- 19131554 TI - Expression of the oncofetal ED-B-containing fibronectin isoform in hematologic tumors enables ED-B-targeted 131I-L19SIP radioimmunotherapy in Hodgkin lymphoma patients. AB - Current treatment of hematologic malignancies involves rather unspecific chemotherapy, frequently resulting in severe adverse events. Thus, modern clinical research focuses on compounds able to discriminate malignant from normal tissues. Being expressed in newly formed blood vessels of solid cancers but not in normal mature tissues, the extradomain B of fibronectin (ED-B FN) is a promising target for selective cancer therapies. Using immunohistology with a new epitope retrieval technique for paraffin-embedded tissues, ED-B FN expression was found in biopsies from more than 200 Hodgkin and non-Hodgkin lymphoma patients of nearly all entities, and in patients with myeloproliferative diseases. ED-B FN expression was nearly absent in normal lymph nodes (n = 10) and bone marrow biopsies (n = 9). The extent of vascular ED-B FN expression in lymphoma tissues was positively correlated with grade of malignancy. ED-B FN expression was enhanced in lymph nodes with severe lymphadenopathy and in some hyperplastic tonsils. The in vivo accessibility of ED-B FN was confirmed in 3 lymphoma patients, in whom the lymphoma lesions were visualized on scintigraphy with (131)I-labeled L19 small immunoprotein ((131)I-L19SIP). In 2 relapsed Hodgkin lymphoma patients(131)I-L19SIP radioimmunotherapy induced a sustained partial response, qualifying ED-B FN as a promising target for antibody-based lymphoma therapies. PMID- 19131553 TI - Gene expression profiling of ATL patients: compilation of disease-related genes and evidence for TCF4 involvement in BIRC5 gene expression and cell viability. AB - Adult T-cell leukemia/lymphoma (ATL) is an aggressive and fatal disease. We have examined 32 patients with smoldering, chronic, lymphoma and acute leukemia using Affymetrix HG-U133A2.0 arrays. Using the BRB array program, we identified genes differentially expressed in leukemia cells compared with normal lymphocytes. Several unique genes were identified that were overexpressed in leukemic cells, including TNFSF11, RGS13, MAFb, CSPG2, C/EBP-alpha, and TCF4; 200 of the most highly overexpressed ATL genes were analyzed by the Pathway Studio, version 4.0 program. ATL leukemia cells were characterized by an increase in genes linked to "central" genes CDC2/cyclin B1, SYK/LYN, proliferating cell nuclear antigen, and BIRC5. Because of its potential therapeutic importance, we focused our studies on the regulation and function of BIRC5, whose expression was increased in 13 of 14 leukemia samples. TCF4 reporter assays and transfection of DN-TCF4 demonstrated that TCF4 regulates BIRC5 gene expression. Functionally, transfection of ATL cells with BIRC5 shRNA decreased BIRC5 expression and cell viability 80%. Clinical treatment of ATL patients with Zenapax or bortezomib decreased BIRC5 expression and cell viability. These experiments represent the first direct experimental evidence that BIRC5 plays an important role in ATL cell viability and provides important insight into ATL genesis and potential targeted therapies. PMID- 19131555 TI - Essential role of spi-1-like (spi-1l) in zebrafish myeloid cell differentiation. AB - The ETS protein Spi-1/Pu.1 plays a pivotal and widespread role throughout hematopoiesis in many species. This study describes the identification, characterization, and functional analysis of a new zebrafish spi transcription factor spi-1-like (spi-1l) that is expressed in primitive myeloid cells, erythro myelo progenitor cells, and in the adult kidney. Spi-1l functions genetically downstream of etsrp, scl, and spi-1/pu.1 in myeloid differentiation. Spi-1l is coexpressed in a subset of spi-1/pu.1 cells and its function is necessary and sufficient for macrophage and granulocyte differentiation. These results establish a critical role for spi-1l in zebrafish myeloid cell differentiation. PMID- 19131556 TI - TLR-mediated immune activation in HIV. PMID- 19131557 TI - Ironing out complementary medicine. PMID- 19131558 TI - Graft-versus-host disease: proteomics comes of age. PMID- 19131560 TI - Novel epididymal proteins as targets for the development of post-testicular male contraception. AB - Apart from condoms and vasectomy, modern contraceptive methods for men are still not available. Besides hormonal approaches to stop testicular sperm production, the post-meiotic blockage of epididymal sperm maturation carries lots of promise. Microarray and proteomics techniques and libraries of expressed sequence tags, in combination with digital differential display tools and publicly available gene expression databases, are being currently used to identify and characterize novel epididymal proteins as putative targets for male contraception. The data reported indicate that these technologies provide complementary information for the identification of novel highly expressed genes in the epididymis. Deleting the gene of interest by targeted ablation technology in mice or using immunization against the cognate protein are the two preferred methods to functionally validate the function of novel genes in vivo. In this review, we summarize the current knowledge of several epididymal proteins shown either in vivo or in vitro to be involved in the epididymal sperm maturation. These proteins include CRISP1, SPAG11e, DEFB126, carbonyl reductase P34H, CD52, and GPR64. In addition, we introduce novel proteinases and protease inhibitor gene families with potentially important roles in regulating the sperm maturation process. Furthermore, potential contraceptive strategies as well as delivery methods will be discussed. Despite the progress made in recent years, further studies are needed to reveal further details in the epididymal sperm maturation process and the factors involved, in order to facilitate the development of new epididymal contraceptives. PMID- 19131559 TI - Dasatinib induces a response in chronic lymphocytic leukemia. PMID- 19131561 TI - Between the professional and the private: the meaning of working with intimate partner violence in social workers' private lives. AB - This qualitative study examines the impact of working with intimate partner violence on therapists' marital relationships and gender identity. Data were collected by in-depth semistructured interviews with 14 experienced women social workers working in domestic violence treatment centers in Israel. Findings indicate that the boundaries between workers' private and professional lives are blurred and work experiences influence their intimate relationships and gender role identities, leading to overall questioning of their relationships. This newly rediscovered consciousness reshapes the meaning of workers' couple relationships. Such shift between private and professional should be considered when training workers to intervene with intimate partner violence. PMID- 19131563 TI - Bile Acid sulfation: a pathway of bile acid elimination and detoxification. AB - Sulfotransferase-2A1 catalyzes the formation of bile acid-sulfates (BA-sulfates). Sulfation of BAs increases their solubility, decreases their intestinal absorption, and enhances their fecal and urinary excretion. BA-sulfates are also less toxic than their unsulfated counterparts. Therefore, sulfation is an important detoxification pathway of BAs. Major species differences in BA sulfation exist. In humans, only a small proportion of BAs in bile and serum are sulfated, whereas more than 70% of BAs in urine are sulfated, indicating their efficient elimination in urine. The formation of BA-sulfates increases during cholestatic diseases. Therefore, sulfation may play an important role in maintaining BA homeostasis under pathologic conditions. Farnesoid X receptor, pregnane X receptor, constitutive androstane receptor, and vitamin D receptor are potential nuclear receptors that may be involved in the regulation of BA sulfation. This review highlights current knowledge about the enzymes and transporters involved in the formation and elimination of BA-sulfates, the effect of sulfation on the pharmacologic and toxicologic properties of BAs, the role of BA sulfation in cholestatic diseases, and the regulation of BA sulfation. PMID- 19131562 TI - Biomarkers of human exposure to acrylamide and relation to polymorphisms in metabolizing genes. AB - Acrylamide (AA) is formed in heat treated carbohydrate rich foods in the so called Maillard reaction. AA is readily absorbed in the body and converted to glycidamide (GA) by epoxidation by the CYP2E1 (cytochrome P450 2E) enzyme. Both AA and GA may be detoxified through direct conjunction to glutathione by glutathione-S-transferases and GA by hydrolysis to glyceramide. Recently, we reported that biomarkers of AA exposure reflect intake of major food sources of AA; there were large interindividual variations in the blood ratio of GA-Hb/AA-Hb (GA- and AA-hemoglobin adducts). In this study we investigated whether the ratio of GA-Hb/AA-Hb in subjects could be related to polymorphic differences in genes coding for metabolizing enzymes CYP2E1, EPHX1 (microsomal epoxide hydrolase), GSTM1, GSTT1, and GSTP1, all being expected to be involved in the activation and detoxification of AA-associated adducts. We found significant associations between GSTM1 and GSTT1 genotypes and the ratio of GA-Hb/AA-Hb (p = 0.039 and p = 0.006, respectively). The ratio of GA-Hb/AA-Hb in individuals with the combined GSTM1- and GSTT1-null variants was significantly (p = 0.029) higher than those with the wild-type genotypes. Although the number of subjects was small, there were also significant associations with other combinations; CYP2E1 (Val179Val) plus GSTM1-null (p = 0.022); CYP2E1 (Val/Val), GSTM1-null plus GSTT1-null (p = 0.047); and CYP2E1 (Val/Val), GSTT1 null, EPHX1 (Tyr113Tyr) plus EPHX1 (His139Arg) (p = 0.018). Individuals with these combined genotypes had significantly higher blood ratio of GA-Hb/AA-Hb than other combinations. The observed associations correspond with what would be expected from the relative roles of these enzymes in activation and detoxification of AA, except for individuals with the EPHX1 (His139Arg) variant. The internal dose of genotoxic metabolite and also the concentration of AA in blood seem to be affected by these polymorphic genes. The genotypes and their combination may constitute useful biomarkers for the assessment of individual susceptibility to AA intake, and could add to the precision of epidemiological studies of dietary cancer. PMID- 19131564 TI - The differential association of kidney dysfunction with small and large arterial elasticity: the multiethnic study of atherosclerosis. AB - Vascular remodeling may be a mechanism linking chronic kidney disease to cardiovascular disease. Whether early kidney dysfunction is associated with small and large arterial remodeling is not well understood. Using multivariable linear regression, back-transforming beta-coefficients to relative difference, the authors studied the association of cystatin C, creatinine-based estimated glomerular filtration rate (GFR), and albuminuria with small (SAE) and large (LAE) arterial elasticity and aortic distensibility among 6,282 participants in the Multiethnic Study of Atherosclerosis at baseline (2000-2002). Compared with the lowest quintile, higher quintiles of cystatin C were incrementally associated with lower SAE: third quintile relative difference = -5% (95% confidence interval (CI): -8, -2); fourth quintile relative difference = -10% (95% CI: -13, -8); and highest quintile relative difference = -16% (95% CI: -20, -12). By use of creatinine, the association was observed only among those with chronic kidney disease (estimated GFR, <60 mL/minute/1.73 m(2)): relative difference = -9% (95% CI: -13, -4). Albuminuria was significantly associated with lower SAE: relative difference = -6% (95% CI: -10, -1). Cystatin C was associated with lower LAE only at the highest quintile (relative difference = -3%, 95% CI: -6, 0) compared with the lowest quintile. By use of creatinine, chronic kidney disease was not independently associated with LAE (P = 0.912). Cystatin C, estimated GFR, and albuminuria were not associated with aortic distensibility (P = 0.26, 0.48, 0.45). Early kidney dysfunction is significantly associated with decreased arterial elasticity in smaller arteries and, to a lesser degree, in larger arteries. PMID- 19131565 TI - Is maternal periodontal disease a risk factor for preterm delivery? AB - Several studies have suggested an association between maternal periodontal disease and preterm delivery, but this has not been a consistent finding. In 2006 2007, the authors examined the relation between maternal periodontal disease and preterm delivery among 467 pregnant Thai women who delivered a preterm singleton infant (<37 weeks' gestation) and 467 controls who delivered a singleton infant at term (> or =37 weeks' gestation). Periodontal examinations were performed within 48 hours after delivery. Participants' periodontal health status was classified into 4 categories according to the extent and severity of periodontal disease. Logistic regression was used to estimate odds ratios and 95% confidence intervals. Preterm delivery cases and controls were similar with regard to mean probing depth, mean clinical attachment loss, and mean percentage of sites exhibiting bleeding on probing. After controlling for known confounders, the authors found that severe clinical periodontal disease was not associated with an increased risk of preterm delivery (odds ratio = 1.20, 95% confidence interval: 0.67, 2.16). In addition, there was no evidence of a linear increase in risk of preterm delivery or its subtypes associated with increasing severity of periodontal disease (P(trend) > 0.05). The results of this case-control study do not provide convincing evidence that periodontal disease is associated with preterm delivery or its subtypes among Thai women. PMID- 19131566 TI - Smoking and Parkinson's disease: using parental smoking as a proxy to explore causality. AB - In epidemiologic studies and in studies of discordant twins, cigarette smoking has been consistently associated with a lower risk of Parkinson's disease, but whether this association is causal remains controversial. Alternatively, an infectious or toxic exposure in childhood or early adulthood could affect both the reward mechanisms that determine smoking behavior and the future risk of Parkinson's disease. If so, parental smoking, commonly established before the birth of the first child, would be unlikely to be related to Parkinson's disease risk. The authors assessed the association between Parkinson's disease and parental smoking during childhood in the Nurses' Health Study and the Health Professionals Follow-up Study conducted in the United States. During 26 years and 18 years of follow-up, respectively, 455 newly diagnosed Parkinson's disease cases were documented among those who provided information on parental smoking. The age-adjusted, pooled relative rate of Parkinson's disease was 0.73 (95% confidence interval: 0.53, 1.00; P-trend = 0.04) comparing participants who reported that both parents smoked with those who reported that neither did. Adjustment for caffeine and alcohol intake did not materially change the results. If the inverse association between smoking and Parkinson's disease were due to confounding by an environmental factor or were the result of reverse causation, it is unlikely that parental smoking would predict Parkinson's disease. PMID- 19131567 TI - Views of sexual abuse professionals about sex offender notification policies. AB - This study examines the views of 261 sexual abuse professionals regarding sex crime policies. Few participants believe that community notification is very effective in reducing sex offenses, but about half believe that all sex offenders should be subject to public disclosure. About one quarter of respondents opine that sex offenders should not be allowed to live in their communities, but a majority do not view residential housing restrictions for sex offenders as effective at reducing recidivism. Most believe in the helpfulness of therapeutic intervention. More than half say that they will support sex offender policies even without scientific evidence of their effectiveness. Criminal justice professionals are more likely to approve of these laws compared to mental health professionals. Those with more conservative political leanings also favor more restrictive policies. There are few differences attributable to gender, parenting status, or victimization experiences. PMID- 19131568 TI - Cloning of gonadotropin-releasing hormone I complementary DNAs in songbirds facilitates dissection of mechanisms mediating seasonal changes in reproduction. AB - Temperate zone animals exhibit seasonal variation in reproductive physiology. In most cases, seasonal changes in reproductive states are regulated by changes in GnRH1 secretion, rather than synthesis, from the preoptic area (POA)/anterior hypothalamus. An important exception occurs in some songbirds that become photorefractory to the stimulatory effects of long days and show profound decreases in brain GnRH1 protein content. Whether this decline reflects changes in gene expression is unknown because of past failures to measure GNRH1 mRNA levels, due in large part to the absence of available GNRH1 gene sequence in this taxon. Here, we report the first cloning of GNRH1 cDNAs in two songbirds: European starlings and zebra finches. Consistent with the size of the prepro hormone in other avian and non-avian species, the open-reading frames predict proteins of 91 and 92 amino acids, respectively. Whereas the decapeptide in both species is perfectly conserved with chicken GnRH1, the amino acid identity in the signal peptide and GNRH associated peptide subdomains are significantly less well conserved. At the nucleotide level, the starling and zebra finch coding sequences are approximately 88% identical to each other but only approximately 70% identical to chicken GNRH1. In situ hybridization using radiolabeled cRNA probes demonstrated GNRH1 mRNA expression primarily in the POA, consistent with previous studies on the distribution of the GnRH1-immunoreactive cell bodies. Furthermore, we provide evidence for photoperiod-dependent regulation of GNRH1 mRNA in male starlings. Declines in GNRH1 mRNA levels occur in parallel with testicular involution. Thus, photorefractoriness is associated with decreases in GNRH1 gene expression in the medial POA. PMID- 19131569 TI - Kinetic complexity of the global response to glucocorticoid receptor action. AB - We have characterized the kinetic response of gene targets throughout the murine genome to transcriptional modulation by the glucocorticoid receptor (GR). In contrast to a model in which multiple genes are either repressed or activated during the GR response, the vast majority of responsive genes are subject to complex regulation profiles, frequently with alternate activation and repression phases. We also observe that GR binding at response elements does not always correlate with the target gene response profile. Thus, the cellular response to GR stimulation involves a highly orchestrated series of regulatory actions and not simply a binary response to hormone. PMID- 19131570 TI - Reduced behavioral response to gonadal hormones in mice shipped during the peripubertal/adolescent period. AB - Animals shipped from commercial suppliers to laboratories are exposed to a wide variety of stressors. Female C57Bl6/J mice shipped during the peripubertal/adolescent period (6 wk old) display lower levels of female sexual behavior in response to estradiol and progesterone injections after ovariectomy when tested in adulthood than female mice shipped in adulthood (12 wk old). These shipping-induced reductions in female sexual behavior appear to be limited to a vulnerable period around the time of puberty. Likewise, male mice shipped at 6 wk of age express lower levels of masculine sexual behavior in response to testosterone treatment as adults than do mice shipped when 12 wk old. RIA of corticosterone levels in response to behavior testing revealed that, upon first exposure to testing, mice shipped at 6 wk of age have reduced corticosterone levels. These results suggest that during the peripubertal/adolescent period, mice of both sexes are susceptible to the effects of stressors associated with shipping. Furthermore, they suggest that stress during this period has enduring, negative influences on behavioral responses to estradiol and progesterone in females and to testosterone in males, and it induces changes in response of the hypothalamic-pituitary-adrenal axis. These results suggest that age at shipping is a critical variable that may influence many endocrinological studies, and they suggest that the peripubertal/adolescent period is a period of vulnerability to some stressors. PMID- 19131571 TI - Porcine marginal mass islet autografts resist metabolic failure over time and are enhanced by early treatment with liraglutide. AB - Although insulin independence is maintained in most islet recipients at 1 yr after transplant, extended follow-up has revealed that many patients will eventually require insulin therapy. Previous studies have shown that islet autografts are prone to chronic failure in large animals and humans, suggesting that nonimmunological events contribute to islet graft functional decay. Early intervention with therapies that promote graft stability should provide a measurable benefit over time. In this study, the efficacy of the long-acting glucagon-like peptide-1 analog liraglutide was explored in a porcine marginal mass islet autograft transplant model. Incubation with liraglutide enhanced porcine islet survival and function after prolonged culture. Most vehicle-treated (83%) and liraglutide-treated (80%) animals became insulin independent after islet autotransplantation. Although liraglutide therapy did not improve insulin independence rates or blood glucose levels after transplant, a significant increase in insulin secretion and acute-phase insulin response was observed in treated animals. Surprisingly, no evidence for deterioration of graft function was observed in any of the transplanted animals over more than 18 months of follow-up despite significant weight gain; in fact, an enhanced response to glucose developed over time even in control animals. Histological analysis showed that intraportally transplanted islets remained highly insulin positive, retained alpha-cells, and did not form amyloid deposits. This study demonstrates that marginal mass porcine islet autografts have stable long-term function, even in the presence of an increasing metabolic demand. These results are discrepant with previous large animal studies and suggest that porcine islets may be resistant to metabolic failure. PMID- 19131572 TI - RFamide-related peptide-3, a mammalian gonadotropin-inhibitory hormone ortholog, regulates gonadotropin-releasing hormone neuron firing in the mouse. AB - The recent discovery that an RFamide termed gonadotropin-inhibitory hormone is likely to be a hypophysiotrophic gonadotropin release-inhibiting hormone in birds has generated interest into the role of LPXRFamide neuropeptides in the control of gonadotropin secretion in mammals. Recent immunocytochemical studies in birds and mammals have suggested that neurons expressing the mammalian LPXRFamides, RFamide-related peptides (RFRPs) 1 and 3, may innervate and regulate GnRH neurons directly. We used cell-attached electrophysiology in adult male and female GnRH green fluorescent protein-tagged neurons to examine whether RFRP-3 modulated the electrical excitability of GnRH neurons. RFRP-3 was found to exhibit rapid and repeatable inhibitory effects on the firing rate of 41% of GnRH neurons. A small population of GnRH neurons (12%) increased their firing rate in response to RFRP 3, and the remainder was unaffected. No difference was detected in the RFRP-3 responses of GnRH neurons from male, diestrous, or proestrus female mice. The suppressive effect of RFRP-3 was maintained when amino acid transmission was blocked, suggesting a possible direct effect of RFRP-3 upon GnRH neurons. To evaluate the effects of other RFamide neuropeptides on GnRH neurons, we tested the actions of prolactin-releasing peptide-20 and -31. Neither compounds altered the firing rate of GnRH neurons. These studies demonstrate that RFRP-3 has a likely direct suppressive action on the excitability of GnRH neurons, indicating a role for RFRPs in the regulation of gonadotropin secretion in mammals through modulation of GnRH neuron activity. PMID- 19131573 TI - MicroRNA 18 and 124a down-regulate the glucocorticoid receptor: implications for glucocorticoid responsiveness in the brain. AB - Glucocorticoids (GCs) exert profound effects on a variety of physiological processes, including adaptation to stress, metabolism, immunity, and neuronal development. Cellular responsiveness to GCs depends on numerous factors, including the amount of the glucocorticoid receptor (GR) protein. We tested the hypothesis that micro-RNAs (miRs), a recently discovered group of noncoding RNAs involved in mRNA translation, might control GR activity by reducing GR protein levels in neuronal tissues. We tested a panel of five miRs consisting of 124aa, 328, 524, 22, and 18. We found that miRs 18 and 124a reduced GR-mediated events in addition to decreasing GR protein levels. miR reporter assays revealed binding of miR-124a to the 3' untranslated region of GR. In correspondence, the activation of the GR-responsive gene glucocorticoid-induced leucine zipper was strongly impaired by miR-124a and -18 overexpression. Although miR-18 is expressed widely throughout the body, expression of miR-124a is restricted to the brain. Endogenous miR-124a up-regulation during neuronal differentiation of P19 cells was associated with a decreasing amount of GR protein levels and reduced activity of luciferase reporter constructs bearing GR 3' untranslated regions. Furthermore, we show that miR-124a expression varies over time during the stress hyporesponsive period, a neonatal period when GC signaling is modulated. Our findings demonstrate a potential role for miRs in the regulation of cell type specific responsiveness to GCs, as may occur during critical periods of neuronal development. Ultimately, our results may provide a better understanding of the etiology of stress-related diseases as well as the efficacy of GC therapy. PMID- 19131574 TI - Organizational actions of postnatal estradiol in female sheep treated prenatally with testosterone: programming of prepubertal neuroendocrine function and the onset of puberty. AB - Prenatal testosterone (T) exposure defeminizes reproductive neuroendocrine function in female sheep, although the LH surge dysfunctions are initially less severe in gonadally intact females than in females subject to neonatal ovariectomy and estradiol (E) replacement. Because prepubertal ovarian production of E differs quantitatively and qualitatively from chronic E replacement, we tested the hypothesis that postnatal E exacerbates the consequences of prenatal T on the positive, but not the negative, steroid feedback controls of GnRH secretion. Our approach was to characterize prepubertal sensitivity to E negative feedback, the onset and maintenance of progestagenic cycles, and the LH surge response in ovary intact, prenatally untreated (control), and T-treated (T) sheep that were exposed postnatally to only endogenous E, or exposed to excess E by s.c. implant. Sensitivity to E negative feedback was reduced in T females, but excess postnatal E did not further increase LH pulse frequency. Excess E prevented ovarian cycles in several control females, and increased cycle irregularity in T females. However, the LH surge mechanism was functional in all control females (regardless of postnatal E exposure) and in some T females without excess E, but nonfunctional in T females with excess E. These findings suggest that postnatal E does not program increased resistance to E negative feedback, but excess postnatal E does disrupt other mechanisms required for ovarian cyclicity. We conclude that in this precocial species, prenatal steroids are sufficient to program controls of tonic LH secretion, but the LH surge mechanism is susceptible to further programming by postnatal E. PMID- 19131575 TI - Roles of testicular orphan nuclear receptors 2 and 4 in early embryonic development and embryonic stem cells. AB - The testicular orphan nuclear receptors (TRs) 2 and 4 act as either transcriptional activators or regulatory proteins of other nuclear receptor superfamily members. With no identified cognate ligands, their physiological roles remain unclear. Here we showed the phenotypes of TR2(-/-):TR4(-/-) mutant embryos, which reveal that the loss of TR2 and TR4 causes early embryonic lethality and increased cell death. We also found that TR2 and TR4 are expressed in blastocysts and embryonic stem (ES) cells, and can act as transcriptional activators in ES cells. The results on further investigating the roles of TR2 and TR4 in ES cells showed that TR2 and TR4 were differentially expressed when ES cells were induced into different specialized cell types, and their expression is regulated by retinoic acid. Knocking down TR2 and TR4 mRNAs decreased the expression of Oct-3/4 and Nanog genes. Mechanism dissection suggests that TR2 and TR4 may affect the Oct-3/4 gene by binding to a direct repeat-1 element located in its promoter region, which is influenced by retinoic acid. Together, our findings highlight possible roles for TR2 and TR4 in early embryonic development by regulating key genes involved in stem cell self-renewal, commitment, and differentiation. PMID- 19131576 TI - New insights into the role of androgens in wolffian duct stabilization in male and female rodents. AB - Androgen-mediated wolffian duct (WD) development is programmed between embryonic d 15.5 (e15.5) and 17.5 in male rats, and WD differentiation has been shown to be more susceptible to reduced androgen action than is its initial stabilization. We investigated regulation of these events by comparing fetal WD development at e15.5-postnatal d0 in male and female androgen receptor knockout mice, and in rats treated from e14.5 with flutamide (100 mg/kg/d) plus di-n(butyl) phthalate (500 mg/kg/d) to block both androgen action and production, testosterone propionate (20 mg/kg/d) to masculinize females, or vehicle control. In normal females, WD regression occurred by e15.5 in mice and e18.5 in rats, associated with a lack of epithelial cell proliferation and increased apoptosis, disintegration of the basement membrane, and reduced epithelial cell height. Exposure to testosterone masculinized female rats including stabilization and partial differentiation of WDs. Genetic or chemical ablation of androgen action in males prevented masculinization and induced WD regression via similar processes to those in normal females, except this occurred 2-3 d later than in females. These findings provide the first evidence that androgens may not be the only factor involved in determining WD fate. Other factors may promote survival of the WD in males or actively promote WD regression in females, suggesting sexually dimorphic differences in the preprogrammed setup of the WD. PMID- 19131577 TI - Connection of left corticospinal tract and broca's area in a patient with intracerebral hemorrhage. PMID- 19131578 TI - Forced, not voluntary, exercise improves motor function in Parkinson's disease patients. AB - BACKGROUND: Animal studies indicate forced exercise (FE) improves overall motor function in Parkinsonian rodents. Global improvements in motor function following voluntary exercise (VE) are not widely reported in human Parkinson's disease (PD) patients. OBJECTIVE: The aim of this study was to compare the effects of VE and FE on PD symptoms, motor function, and bimanual dexterity. METHODS: Ten patients with mild to moderate PD were randomly assigned to complete 8 weeks of FE or VE. With the assistance of a trainer, patients in the FE group pedaled at a rate 30% greater than their preferred voluntary rate, whereas patients in the VE group pedaled at their preferred rate. Aerobic intensity for both groups was identical, 60% to 80% of their individualized training heart rate. RESULTS: Aerobic fitness improved for both groups. Following FE, Unified Parkinson's Disease Rating Scale (UPDRS) motor scores improved 35%, whereas patients completing VE did not exhibit any improvement. The control and coordination of grasping forces during the performance of a functional bimanual dexterity task improved significantly for patients in the FE group, whereas no changes in motor performance were observed following VE. Improvements in clinical measures of rigidity and bradykinesia and biomechanical measures of bimanual dexterity were maintained 4 weeks after FE cessation. CONCLUSIONS: Aerobic fitness can be improved in PD patients following both VE and FE interventions. However, only FE results in significant improvements in motor function and bimanual dexterity. Biomechanical data indicate that FE leads to a shift in motor control strategy, from feedback to a greater reliance on feedforward processes, which suggests FE may be altering central motor control processes. PMID- 19131579 TI - Is the ebb of asthma due to the decline of allergic asthma? A prospective consultation-based study by the Swiss Sentinel Surveillance Network, 1999-2005. AB - OBJECTIVE: There are conflicting views on time trends of asthma and atopy during the last 10-15 years. Additional confusion is caused by the term of asthma which is a unifying name for different phenotypes. Asthma has been a topic for investigation to the Swiss Sentinel Surveillance Network (SSSN) since 1989. The objective of the actual study was to determine the influence of the allergic and non-allergic components of asthma on time trends from 1999 to 2005. METHODS: Primary care physicians participating in the SSSN were guided by diagnostic criteria for asthma. Rates of asthma episodes per 1000 consultations were calculated for all, for first and subsequent asthma episodes and for allergic and non-allergic asthma. Allergic asthma was defined as asthmatic manifestations in conjunction with eczema and/or hay fever. The smoothed time trend and its 95% confidence intervals were determined using generalized additive models with a loess smoother adjusting for seasonality. RESULTS: Consultations for allergic asthma have decreased between 1999 and 2005. Looking at different age groups, asthma associated with hay fever was reported with decreasing frequency in all age groups, whereas when associated with eczema, the other used marker of allergic asthma, slightly increased among young children. CONCLUSIONS: The decrease of consultations for asthma is most probably due to the allergic component of asthma. Diagnostic shift over time and ready available medications probably contributed to this phenomenon. Our findings indicate a real decline of allergic asthma. Practice recommendations. Consultations for asthma have decreased over recent years. The decrease of consultations for asthma is most probably due to the allergic component of asthma. Diagnostic shift over time and available medications probably contributed to this phenomenon. Strength of Recommendation Taxonomy (SORT) rating: strength of recommendation = C, level of evidence = 3. PMID- 19131580 TI - An overview of training curricula for diabetes peer educators. AB - Global community members who experience similar health problems gravitate to each other for information and support. Peers may be more approachable and can relate to the particular living circumstances one experiences. In well-resourced countries, people have opportunities for learning diabetes self-management; however, empathy may be more helpful when practical barriers arise. Little is published in medical literature about how to foster diabetes peer support and what is published is often limited to English language. Among those programs available, commonalities are readily seen. There is significant evidence that well-informed people cope better with adapting their lifestyle to medical regimens. Professionally delivered diabetes education has been well defined, but there may be additional benefit from learning from those who are living the experience everyday regarding how to navigate health care systems, handle finances, deal with natural emotions or family relations. Diabetes is epidemic and worldwide. There will never be sufficient traditional health care services to meet all future patients needs. While we persist in training health care professionals to deliver better diabetes care, we can explore how to mobilize willing volunteers to provide additional ongoing support to people with diabetes, where they live and work. While the characteristics of a peer educator have been defined slightly differently by several programs, there is agreement across programs that they need to be able to communicate clearly, they need to be willing to learn, they need to have confidence and they need to be flexible and dependable. PMID- 19131581 TI - ACCF/SCAI/STS/AATS/AHA/ASNC 2009 Appropriateness Criteria for Coronary Revascularization: A Report of the American College of Cardiology Foundation Appropriateness Criteria Task Force, Society for Cardiovascular Angiography and Interventions, Society of Thoracic Surgeons, American Association for Thoracic Surgery, American Heart Association, and the American Society of Nuclear Cardiology: Endorsed by the American Society of Echocardiography, the Heart Failure Society of America, and the Society of Cardiovascular Computed Tomography. AB - The American College of Cardiology Foundation (ACCF), Society for Cardiovascular Angiography and Interventions, Society of Thoracic Surgeons, and the American Association for Thoracic Surgery, along with key specialty and subspecialty societies, conducted an appropriateness review of common clinical scenarios in which coronary revascularization is frequently considered. The clinical scenarios were developed to mimic common situations encountered in everyday practice and included information on symptom status, extent of medical therapy, risk level as assessed by noninvasive testing, and coronary anatomy. Approximately 180 clinical scenarios were developed by a writing committee and scored by a separate technical panel on a scale of 1 to 9. Scores of 7 to 9 indicate that revascularization was considered appropriate and likely to improve health outcomes or survival. Scores of 1 to 3 indicate revascularization was considered inappropriate and unlikely to improve health outcomes or survival. The mid range (4 to 6) indicates a clinical scenario for which the likelihood that coronary revascularization would improve health outcomes or survival was considered uncertain. For the majority of the clinical scenarios, the panel only considered the appropriateness of revascularization irrespective of whether this was accomplished by percutaneous coronary intervention (PCI) or coronary artery bypass graft surgery (CABG). In a select subgroup of clinical scenarios in which revascularization is generally considered appropriate, the appropriateness of PCI and CABG individually as the primary mode of revascularization was considered. In general, the use of coronary revascularization for patients with acute coronary syndromes and combinations of significant symptoms and/or ischemia was viewed favorably. In contrast, revascularization of asymptomatic patients or patients with low-risk findings on noninvasive testing and minimal medical therapy were viewed less favorably. It is anticipated that these results will have an impact on physician decision making and patient education regarding expected benefits from revascularization and will help guide future research. PMID- 19131582 TI - Therapeutic effect of blocking CXCR2 on neutrophil recruitment and dextran sodium sulfate-induced colitis. AB - Dextran sodium sulfate (DSS)-induced colitis in mice is characterized by polymorphonuclear neutrophil (PMN) infiltration into the colonic mucosa and lumen. The mechanism by which this occurs is unclear. To begin to understand the mechanism, we determined the role of the PMN chemokine receptor, CXCR2, in DSS induced colitis by using CXCR2(-/-) mice or by neutralizing CXCR2. DSS was administered through drinking water to CXCR2(-/-) and BALB/c mice for 5 days followed by regular water for 1 day. In the neutralization study, mice were injected with control serum or goat anti-CXCR2 antiserum. BALB/c mice receiving DSS and control serum-injected mice receiving DSS lost weight and showed considerable clinical illness. Histological observation revealed submucosal edema, PMN infiltration into the submucosa and mucosa, extensive crypt damage with abscesses, and ulceration. In contrast, both the CXCR2(-/-) and anti-CXCR2 antiserum-treated mice gained weight and had significantly lower symptom scores. Histology of these mice showed submucosal edema but relatively intact crypt architecture and very few ulcers. Significantly fewer PMNs were found in the mucosa in anti-CXCR2 anti-serum compared with control serum-injected inflamed mice, but no significant difference in eosinophil infiltration was observed between the groups. Our experiments identify a role for CXCR2 in DSS-induced colitis and suggest that antagonizing CXCR2 provides some therapeutic efficacy, possibly by impeding PMN recruitment into the mucosa. Antagonizing CXCR2 may form the basis for therapeutic drugs directed at controlling colitis. PMID- 19131583 TI - The microtubule-targeting agents, PBOX-6 [pyrrolobenzoxazepine 7 [(dimethylcarbamoyl)oxy]-6-(2-naphthyl)pyrrolo-[2,1-d] (1,5)-benzoxazepine] and paclitaxel, induce nucleocytoplasmic redistribution of the peptidyl-prolyl isomerases, cyclophilin A and pin1, in malignant hematopoietic cells. AB - Microtubule assembly and disassembly is required for the maintenance of cell structure, mobility, and division. However, the cellular and biochemical implications of microtubule disruption are not fully understood. Using a proteomic approach, we found that the peptidyl-prolyl isomerase, cyclophilin A, was increased in plasma membrane extracts from chronic myeloid leukemia cells after microtubule disruption. In addition, we found that two peptidyl-prolyl isomerases, cyclophilin A and pin1, are overexpressed up to 10-fold in hematological malignancies compared with normal peripheral blood mononuclear cells. Although previous reports suggest that cyclophilin A is localized to the cytosol of mammalian cells, we found that cyclophilin A and pin1 are both localized to the nucleus and nuclear domains in hematopoietic cells. Microtubule disruption of hematopoietic cells caused a dramatic subcellular redistribution of cyclophilin A and pin1 from the nucleus to the cytosol and plasma membrane. We suggest that this accounts for the increased cyclophilin A at the plasma membrane of chronic myeloid leukemia cells after microtubule disruption. The subcellular redistribution of cyclophilin A and pin1 occurred in a c-Jun NH(2)-terminal kinase- and serine protease-dependent manner. Moreover, the altered subcellular localization of the peptidyl-prolyl isomerases occurred in a dose- and time dependent manner after microtubule disruption and was found to correlate with G(2)/M arrest and precede induced cell death. These results suggest that the function of peptidyl-prolyl isomerases may be influenced by microtubule dynamics throughout the cell cycle, and their altered localization may be an important part of the mechanism by which microtubule-disrupting agents exert their cytostatic effects. PMID- 19131584 TI - Role of the endoplasmic reticulum and mitochondria on quantal catecholamine release from chromaffin cells of control and hypertensive rats. AB - Here, we present the first study on the effects of compounds that interfere with calcium (Ca(2+)) handling by the endoplasmic reticulum (ER) and mitochondria on amperometrically measured quantal catecholamine release from single adrenal chromaffin cells of control and spontaneously hypertensive rats (SHRs). Acetylcholine (ACh) or K(+) pulses triggered spike bursts of secretion by Ca(2+) entry through Ca(2+) channels. ER Ca(2+) release triggered by a mixture of caffeine, ryanodine, and thapsigargin (CRT) or carbonyl cyanide p trifluoromethoxyphenylhydrazone (FCCP) (a mitochondrial protonophore) also caused bursts of secretory spikes. The spike bursts generated by ACh, K(+), CRT, and FCCP were 3 to 4 times longer in SHRs compared with control cells; furthermore, the individual spikes were faster and had 3-fold greater quantal size. In additional experiments, a 90-s treatment was made with CRT or FCCP to block Ca(2+) handling by the ER and mitochondria. In these conditions, the integrated spike burst responses elicited by ACh and K(+) were potentiated 2- to 3-fold in control and SHR cells. This suggests that variations in Ca(2+) entry and its subsequent redistribution into the ER and mitochondria are not responsible for the greater secretion seen in SHRs compared with control cells; rather, such differences seem to be due to greater quantal content of spike bursts and to greater quantal size of individual amperometric events. PMID- 19131586 TI - A 17-year-old male with pseudotumor cerebri secondary to performance-enhancing steroids triggering venous thrombosis in the brain. AB - This article is a case report of a 17-year-old male who presented with a headache and blurry vision. He subsequently was noted to have papilledema on a fundoscopic examination and an initial normal magnetic resonance imaging and computed tomography of his head; his condition was, therefore, diagnosed as pseudotumor cerebri. A subsequent magnetic resonance venography of his head revealed venous thrombosis, and other investigations revealed an elevated factor VIII level as well as a mutation at the MTHFR locus, consistent with an elevated risk for hypercoagulability. In addition, he admitted to steroid usage for purposes of performance enhancement in baseball. The patient's condition eventually improved with acetazolamide and serial lumbar punctures. Steroids have been linked to predisposition to hypercoagulable states, but there are no reports identified by these authors that link performance-enhancing steroids with pseudotumor cerebri as a result of a coagulation dyscrasia. PMID- 19131585 TI - Mitochondria-derived reactive oxygen species mediate heme oxygenase-1 expression in sheared endothelial cells. AB - Bovine aortic endothelial cells (ECs) respond to nitric oxide (NO) donors by activating the redox-sensitive NF-E2-related factor 2/antioxidant response element pathway and up-regulating heme oxygenase (HO)-1 expression. EC exposure to steady laminar shear stress causes a sustained increase in NO, a transient increase in reactive oxygen species (ROS), and activation of the HO-1 gene. Because steady laminar flow increases the mitochondrial superoxide (O(2)(*-)) production, we hypothesized that mitochondria-derived ROS play a role in shear induced HO-1 expression. Flow (10 dynes/cm(2), 6 h)-induced expression of HO-1 protein was abolished when BAECs were preincubated and sheared in the presence of either N(G)-nitro-L-arginine methyl ester or N-acetyl-L-cysteine, suggesting that either NO or ROS up-regulates HO-1. Ebselen and diphenylene iodonium blocked HO-1 expression, and uric acid had no effect. The mitochondrial electron transport chain inhibitors, myxothiazol, rotenone, or antimycin A, and the mitochondria targeted antioxidant peptide, Szeto-Schiller (SS)-31, which scavenges O(2)(*-), hydrogen peroxide (H(2)O(2)), peroxynitrite, and hydroxyl radicals, markedly inhibited the increase in HO-1 expression. These data collectively suggest that mitochondrial H(2)O(2) mediates the HO-1 induction. MitoSOX and 2',7' dichlorofluorescin (DCF) fluorescence showed that mitochondrial O(2)(*-) levels and intracellular peroxides, respectively, are higher in sheared ECs compared with static controls and, in part, dependent on NO. SS-31 significantly inhibited both the shear-induced MitoSOX and DCF fluorescence signals. Either phosphatidylinositol 3-kinase or mitogen-activated protein kinase cascade inhibitors blocked the HO-1 induction. In conclusion, under shear, EC mitochondria-derived H(2)O(2) diffuses to the cytosol, where it initiates oxidative signaling leading to HO-1 up-regulation and maintenance of the atheroprotective EC status. PMID- 19131587 TI - Childhood hepatitis A virus infection complicated by Bell's palsy. PMID- 19131588 TI - Bone marrow-derived cells are not the origin of the cancer stem cells in ultraviolet-induced skin cancer. AB - Several lines of evidence have demonstrated that various cancers are derived from cancer stem cells (CSCs), which are thought to originate from either tissue stem or progenitor cells. However, recent studies have suggested that the origin of CSCs could be bone marrow-derived cells (BMDCs); for example, gastric cancer, which follows persistent gastric inflammation, appears to originate from BMDCs. Although our previous research showed the capability of BMDCs to differentiate into epidermal keratinocytes, it has yet to be determined whether skin CSCs originate from BMDCs. To assess the possibility that BMDCs could be the origin of CSCs in skin squamous cell carcinoma (SCC), we used a mouse model of UVB-induced skin SCC. We detected a low percentage of BMDCs in the lesions of epidermal dysplasia (0.59%), SCC in situ (0.15%), and SCC (0.03%). Furthermore, we could not find any evidence of clonal BMDC expansion. In SCC lesions, we also found that most of the BMDCs were tumor-infiltrating hematopoietic cells. In addition, BMDCs in the SCC lesions lacked characteristics of epidermal stem cells, including expression of stem cell markers (CD34, high alpha6 integrin) and the potential retention of BrdU label. These results indicate that BMDCs are not a major source of malignant keratinocytes in UVB-induced SCC. Therefore, we conclude that BMDCs are not the origin of CSCs in UVB-induced SCC. PMID- 19131589 TI - Proteome-wide identification of novel binding partners to the oncogenic fusion gene protein, NPM-ALK, using tandem affinity purification and mass spectrometry. AB - Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), an oncogenic fusion gene protein that is characteristically found in a subset of anaplastic large cell lymphomas, promotes tumorigenesis through its functional and physical interactions with various biologically important proteins. The identification of these interacting proteins has proven to be useful to further our understanding of NPM-ALK-mediated tumorigenesis. For the first time, we performed a proteome wide identification of NPM-ALK-binding proteins using tandem affinity purification and a highly sensitive mass spectrometric technique. Tandem affinity purification is a recently developed method that carries a lower background and higher sensitivity compared with the conventional immunoprecipitation-based protein purification protocols. The NPM-ALK gene was cloned into an HB-tagged vector and expressed in GP293 cells. Three independent experiments were performed and the reproducibility of the data was 68%. The vast majority of the previously reported NPM-ALK-binding proteins were detected. We also identified proteins that are involved in various cellular processes that were not previously described in association with NPM-ALK, such as MCM6 and MSH2 (DNA repair), Nup98 and importin 8 (subcellular protein transport), Stim1 (calcium signaling), 82Fip (RNA regulation), and BAG2 (proteosome degradation). We believe that these data highlight the functional diversity of NPM-ALK and provide new research directions for the study of the biology of this oncoprotein. PMID- 19131590 TI - T-cell activation leads to reduced collagen maturation in atherosclerotic plaques of Apoe(-/-) mice. AB - Rupture of the collagenous, fibrous cap of an atherosclerotic plaque commonly causes thrombosis. Activated immune cells can secrete mediators that jeopardize the integrity of the fibrous cap. This study aimed to determine the relationship between T-cell-mediated inflammation and collagen turnover in a mouse model of experimental atherosclerosis. Both Apoe(-/-) x CD4dnTbetaRII mice with defective transforming growth factor-beta receptors in T cells (and hence released from tonic suppression of T-cell activation) and lesion size-matched Apoe(-/-) mice were used. Picrosirius red staining showed a lower content of thick mature collagen fibers in lesions of Apoe(-/-) x CD4dnTbetaRII mice, although both groups had similar levels of procollagen type I or III mRNA and total collagen content in lesions. Analysis of both gene expression and protein content showed a significant decrease of lysyl oxidase, the extracellular enzyme needed for collagen cross-linking, in aortas of Apoe(-/-)--CD4dnTbetaRII mice. T-cell-driven inflammation provoked a selective and limited increase in the expression of proteinases that catabolize the extracellular matrix. Atheromata of Apoe(-/-)- CD4dnTbetaRII mice had increased levels of matrix metalloproteinase-13 and cathepsin S mRNAs and of the active form of cathepsin S protein but no increase was detected in collagen fragmentation. Our results suggest that exaggerated T cell-driven inflammation limits collagen maturation in the atherosclerotic plaque while having little effect on collagen degradation. PMID- 19131591 TI - Expression of CD74, the receptor for macrophage migration inhibitory factor, in non-small cell lung cancer. AB - Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine that is overexpressed in lung cancer. The MIF receptor was recently discovered and found to be the invariant chain of the HLA class II molecule, CD74. We hypothesized that the expression of this receptor-ligand pair in lung cancer is associated with the angiogenic activity and level of CXC chemokine expression in human specimens of non-small cell lung cancer. We, therefore, performed immunolocalization of CD74 and compared it with the localization of MIF in non small cell lung cancer to determine their respective locations, as well as the relationship between the co-expression of MIF-CD74 and angiogenic CXC chemokines with tumor angiogenesis. We found intense CD74 expression by immunohistochemistry in 57 of 70 tumors with minimal to no staining in the remaining 13 tumors. Comparing the localization of CD74 with its putative ligand, MIF, we found that CD74 and MIF were co-expressed in tumors in close proximity, and that co expression of the MIF-CD74 pair was associated with both higher levels of tumor associated angiogenic CXC chemokines (ie, the ELR score) and greater vascularity compared with tumors in which MIF-CD74 co-expression was not present. We also found that MIF induced angiogenic CXC chemokine expression in an autocrine manner in vitro, a function that was specifically inhibited by antibodies to CD74. PMID- 19131592 TI - HIN-200 proteins regulate caspase activation in response to foreign cytoplasmic DNA. AB - The mammalian innate immune system is activated by foreign nucleic acids. Detection of double-stranded DNA (dsDNA) in the cytoplasm triggers characteristic antiviral responses and macrophage cell death. Cytoplasmic dsDNA rapidly activated caspase 3 and caspase 1 in bone marrow-derived macrophages. We identified the HIN-200 family member and candidate lupus susceptibility factor, p202, as a dsDNA binding protein that bound stably and rapidly to transfected DNA. Knockdown studies showed p202 to be an inhibitor of DNA-induced caspase activation. Conversely, the related pyrin domain-containing HIN-200 factor, AIM2 (p210), was required for caspase activation by cytoplasmic dsDNA. This work indicates that HIN-200 proteins can act as pattern recognition receptors mediating responses to cytoplasmic dsDNA. PMID- 19131593 TI - Harmonic convergence in the love songs of the dengue vector mosquito. AB - The familiar buzz of flying mosquitoes is an important mating signal, with the fundamental frequency of the female's flight tone signaling her presence. In the yellow fever and dengue vector Aedes aegypti, both sexes interact acoustically by shifting their flight tones to match, resulting in a courtship duet. Matching is made not at the fundamental frequency of 400 hertz (female) or 600 hertz (male) but at a shared harmonic of 1200 hertz, which exceeds the previously known upper limit of hearing in mosquitoes. Physiological recordings from Johnston's organ (the mosquito's "ear") reveal sensitivity up to 2000 hertz, consistent with our observed courtship behavior. These findings revise widely accepted limits of acoustic behavior in mosquitoes. PMID- 19131595 TI - Self-sustained replication of an RNA enzyme. AB - An RNA enzyme that catalyzes the RNA-templated joining of RNA was converted to a format whereby two enzymes catalyze each other's synthesis from a total of four oligonucleotide substrates. These cross-replicating RNA enzymes undergo self sustained exponential amplification in the absence of proteins or other biological materials. Amplification occurs with a doubling time of about 1 hour and can be continued indefinitely. Populations of various cross-replicating enzymes were constructed and allowed to compete for a common pool of substrates, during which recombinant replicators arose and grew to dominate the population. These replicating RNA enzymes can serve as an experimental model of a genetic system. Many such model systems could be constructed, allowing different selective outcomes to be related to the underlying properties of the genetic system. PMID- 19131596 TI - A celebration and a challenge. PMID- 19131594 TI - Function of mitochondrial Stat3 in cellular respiration. AB - Cytokines such as interleukin-6 induce tyrosine and serine phosphorylation of Stat3 that results in activation of Stat3-responsive genes. We provide evidence that Stat3 is present in the mitochondria of cultured cells and primary tissues, including the liver and heart. In Stat3(-/-) cells, the activities of complexes I and II of the electron transport chain (ETC) were significantly decreased. We identified Stat3 mutants that selectively restored the protein's function as a transcription factor or its functions within the ETC. In mice that do not express Stat3 in the heart, there were also selective defects in the activities of complexes I and II of the ETC. These data indicate that Stat3 is required for optimal function of the ETC, which may allow it to orchestrate responses to cellular homeostasis. PMID- 19131597 TI - Conservation. Scientists laud Bush's blue legacy but want more. PMID- 19131598 TI - Climate change. Higher temperatures seen reducing global harvests. PMID- 19131599 TI - U.S. national security. A new spy agency asks academics for help in meeting its mission. PMID- 19131600 TI - Neuroscience. Brain scans of pain raise questions for the law. PMID- 19131601 TI - Microbiology. TB bacteria may reign over cells intended to bridle them. PMID- 19131602 TI - Particle physics. Indian neutrino detector hits snag on environmental concerns. PMID- 19131604 TI - Archaeology. Seeking Africa's first iron men. PMID- 19131603 TI - Evolutionary roots. On the origin of life on Earth. PMID- 19131606 TI - Biomedicine. A discriminating killer. PMID- 19131605 TI - Biomedicine. A new view on--and hope for--an old disease. PMID- 19131608 TI - An award for science is an obsolete notion. PMID- 19131607 TI - Unsung hero Robert C. Gallo. PMID- 19131609 TI - The time to demand funding. PMID- 19131610 TI - Autistic phenotype from MEF2C knockout cells. PMID- 19131611 TI - Science should stick to science. PMID- 19131612 TI - Science careers: where does advocacy fit? PMID- 19131613 TI - Unintended consequences at NIH. PMID- 19131614 TI - Economics. Trade liberalization and economic development. PMID- 19131615 TI - Physics. When infinity does not count. PMID- 19131616 TI - Materials science. Unjamming a polymer glass. PMID- 19131617 TI - Behavior. Surprising emotions. PMID- 19131618 TI - Chemistry. Extending polymer conjugation into the second dimension. PMID- 19131619 TI - Geochemistry. The descent of minerals. PMID- 19131620 TI - Ocean science. Old new nitrogen. PMID- 19131621 TI - Developmental biology. Pluripotent chromatin state. PMID- 19131622 TI - Developmental biology. Histone cross-talk in stem cells. PMID- 19131623 TI - Darwin's Originality. AB - Charles Darwin's theory of natural selection has been hailed as one of the most innovative contributions to modern science. When first proposed in 1859, however, it was widely rejected by his contemporaries, even by those who accepted the general idea of evolution. This article identifies those aspects of Darwin's work that led him to develop this revolutionary theory, including his studies of biogeography and animal breeding, and his recognition of the role played by the struggle for existence. PMID- 19131624 TI - Suppression of metallic conductivity of single-walled carbon nanotubes by cycloaddition reactions. AB - The high carrier mobility of films of semiconducting single-walled carbon nanotubes (SWNTs) is attractive for electronics applications, but the presence of metallic SWNTs leads to high off-currents in transistor applications. The method presented here, cycloaddition of fluorinated olefins, represents an effective approach toward converting the "as grown" commercial SWNT mats into high-mobility semiconducting tubes with high yield and without further need for carbon nanotube separation. Thin-film transistors, fabricated from percolating arrays of functionalized carbon nanotubes, exhibit mobilities >100 square centimeters per volt-second and on-off ratios of 100,000. This method should allow for the use of semiconducting carbon nanotubes in commercial electronic devices and provide a low-cost route to the fabrication of electronic inks. PMID- 19131625 TI - Self-organization of a mesoscale bristle into ordered, hierarchical helical assemblies. AB - Mesoscale hierarchical helical structures with diverse functions are abundant in nature. Here we show how spontaneous helicity can be induced in a synthetic polymeric nanobristle assembling in an evaporating liquid. We use a simple theoretical model to characterize the geometry, stiffness, and surface properties of the pillars that favor the adhesive self-organization of bundles with pillars wound around each other. The process can be controlled to yield highly ordered helical clusters with a unique structural hierarchy that arises from the sequential assembly of self-similar coiled building blocks over multiple length scales. We demonstrate their function in the context of self-assembly into previously unseen structures with uniform, periodic patterns and controlled handedness and as an efficient particle-trapping and adhesive system. PMID- 19131626 TI - Historical warnings of future food insecurity with unprecedented seasonal heat. AB - Higher growing season temperatures can have dramatic impacts on agricultural productivity, farm incomes, and food security. We used observational data and output from 23 global climate models to show a high probability (>90%) that growing season temperatures in the tropics and subtropics by the end of the 21st century will exceed the most extreme seasonal temperatures recorded from 1900 to 2006. In temperate regions, the hottest seasons on record will represent the future norm in many locations. We used historical examples to illustrate the magnitude of damage to food systems caused by extreme seasonal heat and show that these short-run events could become long-term trends without sufficient investments in adaptation. PMID- 19131627 TI - The aryl hydrocarbon nuclear translocator alters CD30-mediated NF-kappaB dependent transcription. AB - Expression and signaling of CD30, a tumor necrosis factor receptor family member, is up-regulated in numerous lymphoid-derived neoplasias, most notably anaplastic large-cell lymphoma (ALCL) and Hodgkin's lymphoma. To gain insight into the mechanism of CD30 signaling, we used an affinity purification strategy that led to the identification of the aryl hydrocarbon receptor nuclear translocator (ARNT) as a CD30-interacting protein that modulated the activity of the RelB subunit of the transcription factor nuclear factor kappaB (NF-kappaB). ALCL cells that were deficient in ARNT exhibited defects in RelB recruitment to NF-kappaB responsive promoters, whereas RelA recruitment to the same sites was potentiated, resulting in the augmented expression of these NF-kappaB-responsive genes. These findings indicate that ARNT functions in concert with RelB in a CD30-induced negative feedback mechanism. PMID- 19131628 TI - HDAC4 regulates neuronal survival in normal and diseased retinas. AB - Histone deacetylase 4 (HDAC4) shuttles between the nucleus and cytoplasm and serves as a nuclear co-repressor that regulates bone and muscle development. We report that HDAC4 regulates the survival of retinal neurons in the mouse in normal and pathological conditions. Reduction in HDAC4 expression during normal retinal development led to apoptosis of rod photoreceptors and bipolar (BP) interneurons, whereas overexpression reduced naturally occurring cell death of the BP cells. HDAC4 overexpression in a mouse model of retinal degeneration prolonged photoreceptor survival. The survival effect was due to the activity of HDAC4 in the cytoplasm and relied at least partly on the activity of hypoxia inducible factor 1alpha (HIF1alpha). These data provide evidence that HDAC4 plays an important role in promoting the survival of retinal neurons. PMID- 19131629 TI - Genetic code supports targeted insertion of two amino acids by one codon. AB - Strict one-to-one correspondence between codons and amino acids is thought to be an essential feature of the genetic code. However, we report that one codon can code for two different amino acids with the choice of the inserted amino acid determined by a specific 3' untranslated region structure and location of the dual-function codon within the messenger RNA (mRNA). We found that the codon UGA specifies insertion of selenocysteine and cysteine in the ciliate Euplotes crassus, that the dual use of this codon can occur even within the same gene, and that the structural arrangements of Euplotes mRNA preserve location-dependent dual function of UGA when expressed in mammalian cells. Thus, the genetic code supports the use of one codon to code for multiple amino acids. PMID- 19131630 TI - tasselseed1 is a lipoxygenase affecting jasmonic acid signaling in sex determination of maize. AB - Sex determination in maize is controlled by a developmental cascade leading to the formation of unisexual florets derived from an initially bisexual floral meristem. Abortion of pistil primordia in staminate florets is controlled by a tasselseed-mediated cell death process. We positionally cloned and characterized the function of the sex determination gene tasselseed1 (ts1). The TS1 protein encodes a plastid-targeted lipoxygenase with predicted 13-lipoxygenase specificity, which suggests that TS1 may be involved in the biosynthesis of the plant hormone jasmonic acid. In the absence of a functional ts1 gene, lipoxygenase activity was missing and endogenous jasmonic acid concentrations were reduced in developing inflorescences. Application of jasmonic acid to developing inflorescences rescued stamen development in mutant ts1 and ts2 inflorescences, revealing a role for jasmonic acid in male flower development in maize. PMID- 19131631 TI - Structure of a type IV secretion system core complex. AB - Type IV secretion systems (T4SSs) are important virulence factors used by Gram negative bacterial pathogens to inject effectors into host cells or to spread plasmids harboring antibiotic resistance genes. We report the 15 angstrom resolution cryo-electron microscopy structure of the core complex of a T4SS. The core complex is composed of three proteins, each present in 14 copies and forming a approximately 1.1-megadalton two-chambered, double membrane-spanning channel. The structure is double-walled, with each component apparently spanning a large part of the channel. The complex is open on the cytoplasmic side and constricted on the extracellular side. Overall, the T4SS core complex structure is different in both architecture and composition from the other known double membrane spanning secretion system that has been structurally characterized. PMID- 19131632 TI - Simpson's paradox in a synthetic microbial system. AB - The maintenance of "public" or "common good" producers is a major question in the evolution of cooperation. Because nonproducers benefit from the shared resource without bearing its cost of production, they may proliferate faster than producers. We established a synthetic microbial system consisting of two Escherichia coli strains of common-good producers and nonproducers. Depending on the population structure, which was varied by forming groups with different initial compositions, an apparently paradoxical situation could be attained in which nonproducers grew faster within each group, yet producers increased overall. We show that a simple way to generate the variance required for this effect is through stochastic fluctuations via population bottlenecks. The synthetic approach described here thus provides a way to study generic mechanisms of natural selection. PMID- 19131633 TI - Mispredicting affective and behavioral responses to racism. AB - Contemporary race relations are marked by an apparent paradox: Overt prejudice is strongly condemned, yet acts of blatant racism still frequently occur. We propose that one reason for this inconsistency is that people misunderstand how they would feel and behave after witnessing racism. The present research demonstrates that although people predicted that they would be very upset by a racist act, when people actually experienced this event they showed relatively little emotional distress. Furthermore, people overestimated the degree to which a racist comment would provoke social rejection of the racist. These findings suggest that racism may persevere in part because people who anticipate feeling upset and believe that they will take action may actually respond with indifference when faced with an act of racism. PMID- 19131635 TI - Identification of amyloidogenic light chains requires the combination of serum free light chain assay with immunofixation of serum and urine. AB - BACKGROUND: The diagnosis of systemic immunoglobulin light-chain (AL) amyloidosis requires demonstration of amyloid deposits in a tissue biopsy and amyloidogenic monoclonal light chains. The optimal strategy to identify the amyloidogenic clone has not been established. We prospectively assessed the diagnostic sensitivity of the serum free light chain (FLC) kappa/lambda ratio, a commercial serum and urine agarose gel electrophoresis immunofixation (IFE), and the high-resolution agarose gel electrophoresis immunofixation (HR-IFE) developed at our referral center in patients with AL amyloidosis, in whom the amyloidogenic light chain was unequivocally identified in the amyloid deposits. METHODS: The amyloidogenic light chain was identified in 121 consecutive patients with AL amyloidosis by immunoelectron microscopy analysis of abdominal fat aspirates and/or organ biopsies. We characterized the monoclonal light chain by using IFE and HR-IFE in serum and urine and the FLC kappa/lambda ratio in serum. We then compared the diagnostic sensitivities of the 3 assays. RESULTS: The HR-IFE of serum and urine identified the amyloidogenic light chain in all 115 patients with a monoclonal gammopathy. Six patients with a biclonal gammopathy were omitted from the statistical analysis. The diagnostic sensitivity of commercial serum and urine IFE was greater than that of the FLC kappa/lambda ratio (96% vs 76%). The combination of serum IFE and the FLC assay detected the amyloidogenic light chain in 96% of patients. The combination of IFE of both serum and urine with the FLC kappa/lambda ratio had a 100% sensitivity. CONCLUSIONS: The identification of amyloidogenic light chains cannot rely on a single test and requires the combination of a commercially available FLC assay with immunofixation of both serum and urine. PMID- 19131636 TI - Multimarker circulating DNA assay for assessing blood of prostate cancer patients. AB - BACKGROUND: Prostate cancer (PCa) detection using serum-based prostate specific antigen (PSA) is limited by frequent false-positive and -negative results. Genetic aberrations such as allelic imbalance (AI) and epigenetic changes such as promoter hypermethylation have been detected in circulating DNA of cancer patients. We hypothesized that circulating multimarker DNA assays detecting both genetic and epigenetic markers in serum would be useful in assessing PCa patients. METHODS: We assayed blood from healthy male donors (n = 40) and 83 patients with American Joint Cancer Committee (AJCC) stage I-IV PCa. DNA was assayed for AI of 6 genome microsatellites. We assessed methylation of RASSF1, RARB2, and GSTP1 using a methylation-specific PCR assay and analyzed the sensitivity of each assay for the detection of genetic or epigenetic changes in circulating DNA. The relation between circulating tumor-related DNA detection and prognostic factors was investigated. RESULTS: The proportion of patients demonstrating AI for > or =1 marker was 47% (38 of 81 patients). Methylation biomarkers were detected in 24 of 83 patients (28%). By combining 2 DNA assays, the number of PCa patients positive for > or =1 methylated or LOH marker increased (52 of 83; 63%). The combined assays detected PCa in 15 of 24 patients (63%) with normal PSA concentrations. The combination of the DNA assays detected the presence of PCa regardless of AJCC stage or PSA concentration. Combination of the DNA and PSA assays gave 89% sensitivity. CONCLUSIONS: This pilot study demonstrates that the combined circulating DNA multimarker assay identifies patients with PCa and may yield information independent of AJCC stage or PSA concentration. PMID- 19131637 TI - Associations of genetic variants in ATP-binding cassette A1 and cholesteryl ester transfer protein and differences in lipoprotein subclasses in the multi-ethnic study of atherosclerosis. AB - BACKGROUND: ATP-binding cassette A1 (ABCA1) and cholesteryl ester transfer protein (CETP) play important roles in the reverse cholesterol transport pathway. The associations of ABCA1 and CETP polymorphisms with lipoprotein subclasses have not been extensively studied. METHODS: We genotyped 2 ABCA1 and 5 CETP polymorphisms in 999 participants of the Multi-Ethnic Study of Atherosclerosis (MESA) and studied their associations with HDL and LDL subclass particle concentrations, measured by nuclear magnetic resonance spectroscopy. RESULTS: ABCA1 and CETP polymorphisms were associated with different and distinct changes in lipoprotein subclass concentrations. The ABCA1 1051G/A AA genotype, previously found to be associated with cardioprotective effects in this cohort, was associated with a 5.5% higher concentration of small HDL particles (P = 0.024). The CETP TaqIB B2B2, -2505C/A AA, and -629C/A AA genotypes, previously demonstrated to lack cardioprotective effects, were associated with 15.2%, 15.4%, and 11.7% higher HDL cholesterol concentrations, respectively, and 36.5%, 40.7%, and 25.4% higher large HDL particle concentrations (P < 0.0001). The minor alleles of the A373P and R451Q polymorphisms were associated with lower large HDL particle concentrations. CONCLUSIONS: Our study of the influence of ABCA1 and CETP genetic variants on lipoprotein subclasses demonstrates the importance of interpreting lipoprotein subclasses within the context of the biochemical processes involved in the alterations. In the case of HDL, the study of subclass particle numbers and sizes may not be sufficiently informative. Assays for HDL function may be needed to supplement quantification of HDL cholesterol and HDL particle numbers and sizes. PMID- 19131638 TI - Transforming growth factor-beta receptor-3 is associated with pulmonary emphysema. AB - Chronic obstructive pulmonary disease (COPD) is a heterogeneous syndrome, including emphysema and airway disease. Phenotypes defined on the basis of chest computed tomography (CT) may decrease disease heterogeneity and aid in the identification of candidate genes for COPD subtypes. To identify these genes, we performed genome-wide linkage analysis in extended pedigrees from the Boston Early-Onset COPD Study, stratified by emphysema status (defined by chest CT scans) of the probands, followed by genetic association analysis of positional candidate genes. A region on chromosome 1p showed strong evidence of linkage to lung function traits in families of emphysema-predominant probands in the stratified analysis (LOD score = 2.99 in families of emphysema-predominant probands versus 1.98 in all families). Association analysis in 949 individuals from 127 early-onset COPD pedigrees revealed association for COPD-related traits with an intronic single-nucleotide polymorphism (SNP) in transforming growth factor-beta receptor-3 (TGFBR3) (P = 0.005). This SNP was significantly associated with COPD affection status comparing 389 cases from the National Emphysema Treatment Trial to 472 control smokers (P = 0.04), and with FEV(1) (P = 0.004) and CT emphysema (P = 0.05) in 3,117 subjects from the International COPD Genetics Network. Gene-level replication of association with lung function was seen in 427 patients with COPD from the Lung Health Study. In conclusion, stratified linkage analysis followed by association testing identified TGFBR3 (betaglycan) as a potential susceptibility gene for COPD. Published human microarray and murine linkage studies have also demonstrated the importance of TGFBR3 in emphysema and lung function, and our group and others have previously found association of COPD-related traits with TGFB1, a ligand for TGFBR3. PMID- 19131639 TI - Alpha(1)-antitrypsin inhibits epithelial Na+ transport in vitro and in vivo. AB - A variety of studies have shown that Na(+) reabsorption across epithelial cells depends on the protease-antiprotease balance. Herein, we investigate the mechanisms by which alpha(1)-antitrypsin (A1AT), a major anti-serine protease in human plasma and lung epithelial fluid and lacking a Kunitz domain, regulates amiloride-sensitive epithelial Na(+) channel (ENaC) function in vitro and in vivo. A1AT (0.05 mg/ml = 1 microM) decreased ENaC currents across Xenopus laevis oocytes injected with human alpha,beta,gamma-ENaC (hENaC) cRNAs, and human lung Clara-like (H441) cells expressing native ENaC, in a partially irreversible fashion. A1AT also decreased ENaC single-channel activity when added in the pipette but not in the bath solutions of ENaC-expressing oocytes patched in the cell-attached mode. Incubation of A1AT with peroxynitrite (ONOO(-)), an oxidizing and nitrating agent, abolished its antiprotease activity and significantly decreased its ability to inhibit ENaC. Intratracheal instillation of normal but not ONOO(-)-treated A1AT (1 microM) in C57BL/6 mice also decreased Na(+) dependent alveolar fluid clearance to the same level as amiloride. Incubation of either H441 cells or ENaC-expressing oocytes with normal but not ONOO(-)-treated A1AT decreased their ability to cleave a substrate of serine proteases. A1AT had no effect on amiloride-sensitive currents of oocytes injected with hENaC bearing Liddle mutations, presumably because these channels remain at the surface longer than the wild-type channels. These data indicate that A1AT may be an important modulator of ENaC activity and of Na(+)-dependent fluid clearance across the distal lung epithelium in vivo by decreasing endogenous protease activity needed to activate silent ENaC. PMID- 19131640 TI - Surfactant-associated protein B is critical to survival in nickel-induced injury in mice. AB - The etiology of acute lung injury is complex and associated with numerous, chemically diverse precipitating factors. During acute lung injury in mice, one key event is epithelial cell injury that leads to reduced surfactant biosynthesis. We have previously reported that transgenic mice that express transforming growth factor alpha (TGFA) in the lung were protected during nickel induced lung injury. Here, we find that the mechanism by which TGFA imparts protection includes maintenance of surfactant-associated protein B (SFTPB) transcript levels and epidermal growth factor receptor-dependent signaling in distal pulmonary epithelial cells. This protection is complex and not accompanied by a diminution in inflammatory mediator transcripts or additional stimulation of antioxidant transcripts. In mouse lung epithelial (MLE-15) cells, microarray analysis demonstrated that nickel increased transcripts of genes enriched in MTF1, E2F-1, and AP-2 transcription factor-binding sites and decreased transcripts of genes enriched in AP-1-binding sites. Nickel also increased Jun transcript and DNA-binding activity, but decreased SFTPB transcript. Expression of SFTPB under the control of a doxycycline-sensitive promoter increased survival during nickel-induced injury as compared with control mice. Together, these findings support the idea that maintenance of SFTPB expression is critical to survival during acute lung injury. PMID- 19131642 TI - A cystic fibrosis respiratory epithelial cell chronically treated by miglustat acquires a non-cystic fibrosis-like phenotype. AB - Cystic fibrosis (CF) is a fatal, autosomal and recessive genetic disease that is mainly due to inactivating mutations in the chloride channel CF transmembrane conductance regulator (CFTR). Sodium hyperabsorption by the airways, profound lung inflammation, and dysregulation of calcium homeostasis, are presumably causally related to loss of CFTR-dependent chloride function in patients with CF. Miglustat (N-butyldeoxynojirimycin, Zavesca), an inhibitor of the alpha-1,2 glucosidase, has been proposed for clinical use in CF because of its effect as a corrector of the defective trafficking of F508del-CFTR. In the present study, we show that daily treatment for 2 months with low concentrations of miglustat on the human CF nasal epithelial cell line, JME/CF15 (F508del/F508del-CFTR), results in progressive, stable, reversible, and sustained correction of F508del-CFTR trafficking, down-regulation of sodium hyperabsorption, and regulation of the calcium homeostasis. In conclusion, we provide here the first evidence that a respiratory CF cell can acquire a non-CF-like phenotype when chronically treated with low concentrations of a pharmacological drug. PMID- 19131641 TI - Participation of mammalian target of rapamycin complex 1 in Toll-like receptor 2- and 4-induced neutrophil activation and acute lung injury. AB - mTOR complex 1 (mTORC1) plays a central role in cell growth and cellular responses to metabolic stress. Although mTORC1 has been shown to be activated after Toll-like receptor (TLR)-4 engagement, there is little information concerning the role that mTORC1 may play in modulating neutrophil function and neutrophil-dependent inflammatory events, such as acute lung injury. To examine these issues, we determined the effects of rapamycin-induced inhibition of mTORC1 on TLR2- and TLR4-induced neutrophil activation. mTORC1 was dose- and time dependently activated in murine bone marrow neutrophils cultured with the TLR4 ligand, LPS, or the TLR2 ligand, Pam(3) Cys-Ser-(Lys)(4) (PAM). Incubation of PAM or LPS-stimulated neutrophils with rapamycin inhibited expression of TNF-alpha and IL-6, but not IkappaB-alpha degradation or nuclear translocation of NF kappaB. Exposure of PAM or LPS-stimulated neutrophils to rapamycin inhibited phosphorylation of serine 276 in the NF-kappaB p65 subunit, a phosphorylation event required for optimal transcriptional activity of NF-kappaB. Rapamycin pretreatment inhibited PAM- or LPS-induced mTORC1 activation in the lungs. Administration of rapamycin also decreased the severity of lung injury after intratracheal LPS or PAM administration, as determined by diminished neutrophil accumulation in the lungs, reduced interstitial pulmonary edema, and diminished levels of TNF-alpha and IL-6 in bronchoalveolar lavage fluid. These results indicate that mTORC1 activation is essential in TLR2- and TLR4-induced neutrophil activation, as well as in the development and severity of acute lung injury. PMID- 19131643 TI - The pro-fibrotic factor IGFBP-5 induces lung fibroblast and mononuclear cell migration. AB - We have previously shown that insulin-like growth factor-binding protein-5 (IGFBP 5) is overexpressed in fibrotic lung tissues and that it induces production of extracellular matrix components such as collagen and fibronectin both in vitro and in vivo. We recently observed mononuclear cell infiltration in lung tissues of mice expressing IGFBP-5. We therefore examined the role of IGFBP-5 on the migration of immune cells. Migration assays demonstrated that IGFBP-5 induced migration of peripheral blood mononuclear cells (PBMCs) in a dose-dependent manner. Preferential migration of monocytes/macrophages, natural killer cells, and T cells was observed. Moreover, the CD4/CD8 ratio of migrating cells was significantly higher in vitro and in vivo in response to IGFBP-5. IGFBP-5 resulted in preferential migration of activated CD4(+) T cells and monocytes. Interestingly, IGFBP-5 also induced migration of primary human lung fibroblasts. Exogenous administration of IGFBP-5 induced activation of mitogen-activated protein kinase (MAPK) signaling cascade but not PI3K in PBMCs. IGFBP-5-induced migration was blocked by the MEK1/2 inhibitor U0126, suggesting that IGFBP-5 induced migration occurs via MAPK activation. Furthermore, monocytes treated with recombinant IGFBP-5 expressed the mesenchymal markers alpha-smooth muscle actin and fibronectin in vitro and in vivo, suggesting that IGFBP-5 can induce the transformation of monocytes into mesenchymal cells. Collectively, our results suggest that IGFBP-5 induces cell migration via MAPK-dependent and IGF-I independent mechanisms. PMID- 19131644 TI - Cigarette smoke alters respiratory syncytial virus-induced apoptosis and replication. AB - Individuals exposed to cigarette smoke have a greater number and severity of viral infections, including respiratory syncytial virus (RSV) infections, than do nonsmokers, but the cellular mechanism is unknown. Our objective was to determine the mechanism by which cigarette smoke augments viral infection. We hypothesize that cigarette smoke causes necrosis and prevents virus-induced cellular apoptosis, and that this is associated with increased inflammation and viral replication. Primary airway epithelial cells were exposed to cigarette smoke extract for 2 days, followed by 1 day of RSV exposure. Western blot detection of cleaved caspases 3 and 7 showed less apoptosis when cells were treated with cigarette smoke before viral infection. This finding was confirmed with ELISA and TUNEL detection of apoptosis. Measures of cell viability, including propidium iodide staining, ATP assay, and cell counts, indicated that cigarette smoke causes necrosis rather than virus-induced apoptosis. Using plaque assay and fluorescently-labeled RSV, we showed that although there were less live cells in the cigarette smoke-pretreated group, viral load was increased. The effect was inhibited by pretreatment of cells with N-acetylcysteine and aldehyde dehydrogenase, suggesting that the effect was primarily mediated by reactive aldehydes. Cigarette smoke causes necrosis rather than apoptosis in viral infection, resulting in increased inflammation and enhanced viral replication. PMID- 19131645 TI - Complement regulator CD59 protects against atherosclerosis by restricting the formation of complement membrane attack complex. AB - Complement is a central effector system within the immune system and is implicated in a range of inflammatory disorders. CD59 is a key regulator of complement membrane attack complex (MAC) assembly. The atherogenic role of terminal complement has long been suspected but is still unclear. Here, we demonstrate that among mice deficient in apolipoprotein (Apo)E, the additional loss of murine CD59 (mCd59ab(-/-)/ApoE(-/-)) accelerated advanced atherosclerosis featuring occlusive coronary atherosclerosis, vulnerable plaque, and premature death and that these effect could be attenuated by overexpression of human CD59 in the endothelium. Complement inhibition using a neutralizing anti-mouse C5 antibody attenuated atherosclerosis in mCd59ab(-/-)/ApoE(-/-) mice. Furthermore, MAC mediated endothelial damage and promoted foam cell formation. These combined results highlight the atherogenic role of MAC and the atheroprotective role of CD59 and suggest that inhibition of MAC formation may provide a therapeutic approach for the treatment of atherosclerosis. PMID- 19131646 TI - ROCK isoform regulation of myosin phosphatase and contractility in vascular smooth muscle cells. AB - Abnormal vascular smooth muscle cell (VSMC) contraction plays an important role in vascular diseases. The RhoA/ROCK signaling pathway is now well recognized to mediate vascular smooth muscle contraction in response to vasoconstrictors by inhibiting myosin phosphatase (MLCP) activity and increasing myosin light chain phosphorylation. Two ROCK isoforms, ROCK1 and ROCK2, are expressed in many tissues, yet the isoform-specific roles of ROCK1 and ROCK2 in vascular smooth muscle and the mechanism of ROCK-mediated regulation of MLCP are not well understood. In this study, ROCK2, but not ROCK1, bound directly to the myosin binding subunit of MLCP, yet both ROCK isoforms regulated MLCP and myosin light chain phosphorylation. Despite that both ROCK1 and ROCK2 regulated MLCP, the ROCK isoforms had distinct and opposing effects on VSMC morphology and ROCK2, but not ROCK1, had a predominant role in VSMC contractility. These data support that although the ROCK isoforms both regulate MLCP and myosin light chain phosphorylation through different mechanisms, they have distinct roles in VSMC function. PMID- 19131647 TI - AMP-activated protein kinase functionally phosphorylates endothelial nitric oxide synthase Ser633. AB - Endothelial nitric oxide synthase (eNOS) plays a central role in maintaining cardiovascular homeostasis by controlling NO bioavailability. The activity of eNOS in vascular endothelial cells (ECs) largely depends on posttranslational modifications, including phosphorylation. Because the activity of AMP-activated protein kinase (AMPK) in ECs can be increased by multiple cardiovascular events, we studied the phosphorylation of eNOS Ser633 by AMPK and examined its functional relevance in the mouse models. Shear stress, atorvastatin, and adiponectin all increased AMPK Thr172 and eNOS Ser633 phosphorylations, which were abolished if AMPK was pharmacologically inhibited or genetically ablated. The constitutively active form of AMPK or an AMPK agonist caused a sustained Ser633 phosphorylation. Expression of gain-/loss-of-function eNOS mutants revealed that Ser633 phosphorylation is important for NO production. The aorta of AMPKalpha2(-/-) mice showed attenuated atorvastatin-induced eNOS phosphorylation. Nano-liquid chromatography/tandem mass spectrometry (LC/MS/MS) confirmed that eNOS Ser633 was able to compete with Ser1177 or acetyl-coenzyme A carboxylase Ser79 for AMPKalpha phosphorylation. Nano-LC/MS/MS confirmed that eNOS purified from AICAR-treated ECs was phosphorylated at both Ser633 and Ser1177. Our results indicate that AMPK phosphorylation of eNOS Ser633 is a functional signaling event for NO bioavailability in ECs. PMID- 19131649 TI - The puzzle of predicting the impact of brain infarcts on cognitive impairment in the aging brain. PMID- 19131648 TI - miR-1 overexpression enhances Ca(2+) release and promotes cardiac arrhythmogenesis by targeting PP2A regulatory subunit B56alpha and causing CaMKII dependent hyperphosphorylation of RyR2. AB - MicroRNAs are small endogenous noncoding RNAs that regulate protein expression by hybridization to imprecise complementary sequences of target mRNAs. Changes in abundance of muscle-specific microRNA, miR-1, have been implicated in cardiac disease, including arrhythmia and heart failure. However, the specific molecular targets and cellular mechanisms involved in the action of miR-1 in the heart are only beginning to emerge. In this study we investigated the effects of increased expression of miR-1 on excitation-contraction coupling and Ca(2+) cycling in rat ventricular myocytes using methods of electrophysiology, Ca(2+) imaging and quantitative immunoblotting. Adenoviral-mediated overexpression of miR-1 in myocytes resulted in a marked increase in the amplitude of the inward Ca(2+) current, flattening of Ca(2+) transients voltage dependence, and enhanced frequency of spontaneous Ca(2+) sparks while reducing the sarcoplasmic reticulum Ca(2+) content as compared with control. In the presence of isoproterenol, rhythmically paced, miR-1-overexpressing myocytes exhibited spontaneous arrhythmogenic oscillations of intracellular Ca(2+), events that occurred rarely in control myocytes under the same conditions. The effects of miR-1 were completely reversed by the CaMKII inhibitor KN93. Although phosphorylation of phospholamban was not altered, miR-1 overexpression increased phosphorylation of the ryanodine receptor (RyR2) at S2814 (Ca(2+)/calmodulin-dependent protein kinase) but not at S2808 (protein kinase A). Overexpression of miR-1 was accompanied by a selective decrease in expression of the protein phosphatase PP2A regulatory subunit B56alpha involved in PP2A targeting to specialized subcellular domains. We conclude that miR-1 enhances cardiac excitation-contraction coupling by selectively increasing phosphorylation of the L-type and RyR2 channels via disrupting localization of PP2A activity to these channels. PMID- 19131650 TI - Outcome of patients with negative CT angiography results for arterial occlusion treated with intravenous thrombolysis. AB - BACKGROUND AND PURPOSE: Stroke patients without evidence of arterial occlusion may not be suitable candidates for thrombolytic therapy. In our study, we investigated the outcomes of patients with negative CT angiography results for arterial occlusion. METHODS: The study included patients treated within 3 hours after symptom onset with intravenous thrombolysis for significant neurological deficit between August 2003 and June 2007. All of the patients were documented with negative CT angiography results for arterial occlusion by independent reviews. Outcome measurements included modified Rankin score at 3 months, incidence of intracranial hemorrhage, and infarction volume on control CT. The predictors of unfavorable outcome (modified Rankin score, 2-6) were identified by multivariate logistic regression. RESULTS: Altogether, 173 patients received intravenous thrombolysis; of those, 138 underwent CT angiography. The CT angiography results were negative for arterial occlusion in 39 (28%) of the patients: mean age, 71+/-10 years; 16 (41%) female; median baseline NIHSS, 11. At 3 months, modified Rankin score of 0 to 1 was achieved in 18 (46%) of the patients; 6 (15%) died; and 3 (8%) had symptomatic parenchymal hemorrhage. The median infarct volume was 1.5 cm(3). The independent predictors of unfavorable clinical outcome were higher age (OR, 1.1; 95% CI, 1.01-1.27), and baseline NIHSS >12 (OR, 18.8; 95% CI, 1.4 to 261). One patient had encephalitis diagnosed. CONCLUSIONS: Negative baseline CT angiography is not uncommon. The risk of intracerebral hemorrhage after thrombolytic therapy for patients without evidence of arterial occlusion is similar to the risk carried in an unselected patient population. Given the prognosis, thrombolytic therapy seems justified; however, etiology other than stroke should be considered. PMID- 19131651 TI - Correlates of common carotid artery lumen diameter in black and white younger adults: the Bogalusa Heart Study. AB - BACKGROUND AND PURPOSE: Vascular remodeling as depicted by increases in arterial lumen diameter occurs in response to development of atherosclerosis. However, data on the correlates of arterial lumen diameter in younger adults by race and sex are limited. METHODS: The study cohort included 734 white and 306 black subjects, aged 20 to 43 years, enrolled in the Bogalusa Heart Study. The common carotid artery lumen diameter at diastole and intima media thickness (IMT) were measured by M- and B-mode ultrasonography, respectively. RESULTS: As a group, blacks versus whites (3.44 mm/m versus 3.37 mm/m, P=0.002) and males versus females (3.45 mm/m versus 3.35 mm/m, P<0.001) had greater height-adjusted lumen diameter. In multivariate analyses of total sample, body mass index, mean arterial pressure, heart rate, and carotid IMT were independent predictors of height-adjusted lumen diameter. The magnitude of the effects of mean arterial pressure and carotid IMT on height-adjusted lumen diameter was significantly different between races with blacks showing greater slopes (difference in slopes: P=0.011 for mean arterial pressure; P=0.033 for IMT); interaction effects of body mass index and mean arterial pressure with sex on height-adjusted lumen diameter were also noted with males showing steeper slopes (difference in slopes: P=0.003 for body mass index; P=0.005 for mean arterial pressure). In addition, the lumen diameter-carotid IMT relationship was stronger in hypertensives than in normotensives (difference in slopes: P=0.038). CONCLUSIONS: Common carotid artery lumen diameter is influenced by carotid artery IMT, adiposity, and blood pressure in a race- and sex-specific manner in asymptomatic younger adults, which may have implications for preventive cardiology. PMID- 19131652 TI - Impaired cerebral vasomotor activity in spontaneous intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Impairment of cerebrovascular autoregulation may promote secondary brain injury in acute brain insults. Until now, only limited data are available on autoregulation in patients with spontaneous intracerebral hemorrhage. In the current study, we aimed to investigate cerebrovascular reactivity and its significance for outcome in spontaneous intracerebral hemorrhage. METHODS: We continuously recorded mean arterial pressure, intracranial pressure, and cerebral perfusion pressure for mean 95 hours in 20 patients with spontaneous intracerebral hemorrhage. The moving correlation coefficient between mean arterial pressure and intracranial pressure (pressure reactivity index), an index of cerebral vasoreactivity, was calculated from the available artifact-free monitoring time (mean, 50.4 hours). RESULTS: In the univariate analysis pressure reactivity index (r=0.66; P=0.002), hemorrhage volume (r=0.62; P=0.007), cerebral perfusion pressure (r=-0.71; P=0.001), mean arterial pressure (r=-0.61; P=0.005), and hematoma growth (r=0.53; P=0.02) significantly correlated with National Institutes of Health Stroke Scale Score at discharge. In a multivariate stepwise linear regression model, pressure reactivity index remained the only independent predictor of outcome (beta=0.659; P=0.004). In the subgroup of patients with pressure reactivity index greater than a functional threshold of >0.2, the correlation between mean cerebral perfusion pressure and outcome remained significant (r=-0.73; P=0.0102), whereas National Institutes of Health Stroke Scale Score at discharge did not correlate with cerebral perfusion pressure in patients with pressure reactivity index <0.2 (r= 0.05; P=0.9078). CONCLUSIONS: We found evidence for impaired cerebral vasomotor activity as measured by pressure reactivity index in patients with spontaneous intracerebral hemorrhage. We suggest that impaired cerebrovascular reactivity contributes to poor outcome in intracerebral hemorrhage patients. This effect may be mediated by fluctuations in cerebral perfusion. PMID- 19131653 TI - Upregulation of macrophage migration inhibitory factor gene expression in stroke. AB - BACKGROUND AND PURPOSE: MIF has been implicated to function in many inflammatory processes. This study examined whether MIF expression was affected in stroke and its underlying molecular mechanism. METHODS: ELISA and qRT-PCR were used to detect MIF protein and mRNA in PBMCs from stroke patients, the ischemic rat brains, and controls. A MIF promoter assay under hypoxia was performed. RESULTS: MIF protein and mRNA were significantly increased in stroke patients. Increasing levels of MIF were correlated to the severity of stroke and peaked 24 hours after stroke. MIF was significantly upregulated in focal ischemic rat brains. The activity of the human MIF promoter was significantly increased under hypoxia compared to normoxia. CONCLUSIONS: MIF gene expression is upregulated after stroke, and hypoxia signaling plays an important role in upregulation of MIF expression under stroke. PMID- 19131654 TI - Cerebral infarcts and cognitive performance: importance of location and number of infarcts. AB - BACKGROUND AND PURPOSE: Cerebral infarcts increase the risk for cognitive impairment. The relevance of location and number of infarcts with respect to cognitive function is less clear. METHODS: We studied the cross-sectional association between number and location of infarcts and cognitive performance in 4030 nondemented participants of the Age Gene/Environment Susceptibility Reykjavik Study. Composite scores for memory, processing speed, and executive function were created from a neuropsychological battery. Subcortical, cortical, and cerebellar infarcts were identified on brain MRI. We performed linear regression analyses adjusted for demographic and vascular risk factors, depression, white matter lesions, and atrophy. RESULTS: Compared to participants with no infarcts, those with infarcts in multiple locations (n=287, 7%) had slower processing speed (beta=-0.19; P<0.001) and poorer memory (beta=-0.16; P<0.001) and executive function (beta=-0.12; P=0.003). Compared to no infarcts, the presence of either subcortical infarcts only (n=275; beta=-0.12; P=0.016) or cortical infarcts only (n=215; beta=-0.17; P=0.001) was associated with poorer memory performance. Compared to no infarcts, a combination of cortical and subcortical infarcts (n=45) was associated with slower processing speed (beta= 0.38; P<0.001) and poorer executive function (beta=-0.22; P=0.02), whereas a combination of cerebellar and subcortical infarcts (n=89) was associated with slower processing speed (beta=-0.15; P=0.04). Infarcts in all 3 locations was associated with slower processing speed (beta=-0.33; P=0.002). CONCLUSIONS: Having infarcts in >1 location is associated with poor performance in memory, processing speed, and executive function, independent of cardiovascular comorbidities, white matter lesions, and brain atrophy, suggesting that both the number and the distribution of infarcts jointly contribute to cognitive impairment. PMID- 19131656 TI - Corticospinal tract pre-wallerian degeneration: a novel outcome predictor for pediatric stroke on acute MRI. AB - BACKGROUND AND PURPOSE: In neonatal arterial ischemic stroke, pre-Wallerian degeneration in descending corticospinal tracts (DCST) on diffusion MRI (DWI) predicts poor outcome. This signal has not been studied in older children. METHODS: A consecutive arterial ischemic stroke cohort (1 month to 18 years) with acute DWI and >12 months of follow-up were enrolled (SickKids Children's Stroke Program). DCST-DWI variables were quantified with a validated software technique and correlations to the Pediatric Stroke Outcome Measure were sought. RESULTS: Abnormal DCST-DWI signal was detected in 20 of 29 children (69%), with 85% having motor deficits on Pediatric Stroke Outcome Measure. DCST variables correlated with hemiparesis included: (1) any abnormal signal within the course of the DCST; (2) midbrain location; (3) percentage of peduncle; (4) vertical length; and (5) relative volume affected (all P<0.003). Unexpectedly, abnormal DWI signal was detected in the contralesional DCST in 7 children, all with severe hemiparesis. DCST signal abnormality increased over time, outlasted infarct DWI changes, and was difficult to appreciate on visual inspection. CONCLUSIONS: DCST-DWI signal is an acute predictor of motor outcome in childhood stroke and can help guide management. Previously unrecognized contralesional DCST signal predicts severe hemiparesis. PMID- 19131655 TI - Extending acute trials to remote populations: a pilot study during interhospital helicopter transfer. AB - BACKGROUND AND PURPOSE: Methods to increase recruitment into acute stroke trials are needed. The purposes of this study were to evaluate the safety and acceptability of initiating acute stroke trials during early helicopter evacuation and to test an intervention to facilitate informed consent. METHODS: A randomized, controlled trial was done with patients with acute stroke who were transferred by helicopter to the University of Iowa Hospitals and Clinics from February 2007 to January 2008. The intervention to be evaluated was the use of fax and a telephone call to the patient/surrogate ahead of helicopter arrival at the outside emergency department. The aim was to improve the rate of subsequent consent (primary outcome) for a pilot trial of a potentially beneficial, low-risk medical intervention (ranitidine) to prevent aspiration pneumonitis. Consenting eligible patients received the infusion during the flight to University of Iowa Hospitals and Clinics. RESULTS: One hundred patients were enrolled. Consent rate was 54% in the intervention group and 50% in the control group (P=0.69). However, the consent rate was higher (69%) when prearrival communications between the coinvestigator and potential subjects were successful (P=0.04). This approach resulted in an average gain of 59 minutes as compared with initiating recruitment on arrival to University of Iowa Hospitals and Clinics. CONCLUSIONS: Enrollment into stroke intervention trials is feasible during helicopter transportation from a community hospital emergency department to a tertiary stroke center. This underused resource may improve trial efficiency by enabling and expediting participation of remote populations currently excluded from research. Consent rates might be further improved by communication strategies that are more successful in reaching patients at outside emergency departments. PMID- 19131658 TI - A population-based study of the prevalence of fatigue after transient ischemic attack and minor stroke. AB - BACKGROUND AND PURPOSE: Fatigue is common after stroke and can be attributable to the increased physical effort associated with severe neurological deficits; however, its presence in those with little motor deficit raises the possibility of confounding by other factors, such as comorbidity, anxiety, and medication. To control for such factors and determine the extent of stroke-specific fatigue, we compared patients with minor stroke who had little or no residual neurological deficit with patients with TIA; both groups had undergone similar investigations and treatment. METHODS: The prevalence of fatigue 6 months after TIA or minor stroke was assessed in consecutive patients using the Chalder fatigue scale in a population-based incidence study (Oxford Vascular Study). Patients were included if they were independent in self-care Barthel Index (>or=18/20) and without major cognitive impairment (Mini-Mental State Examination >or=24/30). Stroke severity at baseline was assessed with the National Institute of Health Stroke Scale (NIHSS). Other potential causes of fatigue were assessed including anxiety, depression, recent life events, medication, and abnormalities in biochemistry or hematologic tests. RESULTS: Seventy-six participants had minor stroke (mean age, 74.1 years; 42 men) and 73 had TIA (mean age, 72.5 years; 40 men). At 6-month follow-up, median Barthel Index score was 20 (interquartile range, 20-20) in both groups. However, fatigue was more common after stroke than TIA (56% vs 29%; OR, 3.14; 95% CI, 1.51-6.57; P=0.0008). This difference was present both in patients with modified Rankin score of 0 at 6 months (23.8% vs 10.3%) and patients with modified Rankin score >or=1 (69.2% vs 48.6%), and remained more frequent in stroke patients after adjustment for potential confounders. Within the group of patients with stroke, the prevalence of fatigue increased with initial stroke severity (87% NIHSS >or=4 vs 48% NIHSS G (additive model; OR, 1.41 with 95% CI, 1.20 to 1.65; P=0.00002) and LTA 26Thr>Asn (OR, 1.19 with 95% CI, 1.06 to 1.34; P=0.003). LTA 252A>G remained significant after adjustment for multiple testing using either the false discovery rate or by permutation testing. The 2 single nucleotide polymorphisms showed no association in hypertensive subjects (eg, LTA 252A>G, OR, 0.93; 95% CI, 0.84 to 1.03; P=0.17). CONCLUSIONS: These observations may indicate an important role of LTA-mediated inflammatory processes in the pathogenesis of ischemic stroke. PMID- 19131665 TI - Direct subunit-dependent multimodal 5-hydroxytryptamine3 receptor antagonism by methadone. AB - Homomeric 5-hydroxytryptamine (5-HT)(3A) and heteromeric 5-HT(3AB) receptors mediate rapid excitatory responses to serotonin in the central and peripheral nervous systems. The alkaloid morphine, in addition to being a mu-opioid receptor agonist, is a potent competitive inhibitor of 5-HT(3) receptors. We examined whether methadone, an opioid often used to treat morphine dependence, also exhibited 5-HT(3) receptor antagonist properties. Racemic (R/S)-methadone inhibited currents mediated by human homomeric 5-HT(3A) receptors (IC(50) = 14.1 +/- 2.5 microM). Incorporation of the 5-HT(3B) subunit into heteromeric 5-HT(3AB) receptors reduced the potency of inhibition by (R/S)-methadone (IC(50) = 41.1 +/- 0.9 microM). (R/S)-Methadone also increased apparent desensitization of both 5 HT(3) receptor subtypes. The inhibition of the 5-HT(3A) receptor was competitive; however, incorporation of the 5-HT(3B) subunit caused the appearance of inhibition that was insurmountable by 5-HT. In the absence of rapid desensitization, when dopamine was used as an agonist of 5-HT(3AB) receptors, the inhibition by (R/S)-methadone was voltage-dependent. The antagonist and desensitization-enhancing effects of (R/S)-methadone were shared by pure (R)- and (S)-methadone enantiomers, which had similar actions on 5-HT-evoked currents mediated by 5-HT(3) receptors. However, (R)-methadone exhibited a larger voltage dependent inhibition of dopamine-evoked currents mediated by 5-HT(3AB) receptors than did (S)-methadone. Inhibition of 5-HT(3A) receptors by (R/S)-methadone was not influenced by voltage. Thus, methadone displays multimodal subunit-dependent antagonism of 5-HT(3) receptors. PMID- 19131666 TI - Fast 3D iterative image reconstruction for SPECT with rotating slat collimators. AB - As an alternative to the use of traditional parallel hole collimators, SPECT imaging can be performed using rotating slat collimators. While maintaining the spatial resolution, a gain in image quality could be expected from the higher photon collection efficiency of this type of collimator. However, the use of iterative methods to do fully three-dimensional (3D) reconstruction is computationally much more expensive and furthermore involves slow convergence compared to a classical SPECT reconstruction. It has been proposed to do 3D reconstruction by splitting the system matrix into two separate matrices, forcing the reconstruction to first estimate the sinograms from the rotating slat SPECT data before estimating the image. While alleviating the computational load by one order of magnitude, this split matrix approach would result in fast computation of the projections in an iterative algorithm, but does not solve the problem of slow convergence. There is thus a need for an algorithm which speeds up convergence while maintaining image quality for rotating slat collimated SPECT cameras. Therefore, we developed a reconstruction algorithm based on the split matrix approach which allows both a fast calculation of the forward and backward projection and a fast convergence. In this work, an algorithm of the maximum likelihood expectation maximization (MLEM) type, obtained from a split system matrix MLEM reconstruction, is proposed as a reconstruction method for rotating slat collimated SPECT data. Here, we compare this new algorithm to the conventional split system matrix MLEM method and to a gold standard fully 3D MLEM reconstruction algorithm on the basis of computational load, convergence and contrast-to-noise. Furthermore, ordered subsets expectation maximization (OSEM) implementations of these three algorithms are compared. Calculation of computational load and convergence for the different algorithms shows a speedup for the new method of 38 and 426 compared to the split matrix MLEM approach and the fully 3D MLEM respectively and a speedup of 16 and 21 compared to the split matrix OSEM and the fully 3D OSEM respectively. A contrast-to-noise study based on simulated data shows that our new approach has comparable accuracy as the fully 3D reconstruction method. The algorithm developed in this study allows iterative image reconstruction of rotating slat collimated SPECT data with equal image quality in a comparable amount of computation time as a classical SPECT reconstruction. PMID- 19131667 TI - Motion correction of PET brain images through deconvolution: II. Practical implementation and algorithm optimization. AB - Image quality is significantly degraded even by small amounts of patient motion in very high-resolution PET scanners. When patient motion is known, deconvolution methods can be used to correct the reconstructed image and reduce motion blur. This paper describes the implementation and optimization of an iterative deconvolution method that uses an ordered subset approach to make it practical and clinically viable. We performed ten separate FDG PET scans using the Hoffman brain phantom and simultaneously measured its motion using the Polaris Vicra tracking system (Northern Digital Inc., Ontario, Canada). The feasibility and effectiveness of the technique was studied by performing scans with different motion and deconvolution parameters. Deconvolution resulted in visually better images and significant improvement as quantified by the Universal Quality Index (UQI) and contrast measures. Finally, the technique was applied to human studies to demonstrate marked improvement. Thus, the deconvolution technique presented here appears promising as a valid alternative to existing motion correction methods for PET. It has the potential for deblurring an image from any modality if the causative motion is known and its effect can be represented in a system matrix. PMID- 19131668 TI - Computerized method for estimation of the location of a lung tumor on EPID cine images without implanted markers in stereotactic body radiotherapy. AB - The purpose of this study was to develop a computerized method for estimation of the location of a lung tumor in cine images on an electronic portal imaging device (EPID) without implanted markers during stereotactic body radiotherapy (SBRT). Each tumor region was segmented in the first EPID cine image, i.e., reference portal image, based on a multiple-gray level thresholding technique and a region growing technique, and then the image including the tumor region was cropped as a 'tumor template' image. The tumor location was determined as the position in which the tumor template image took the maximum cross-correlation value within each consecutive portal image, which was acquired in cine mode on the EPID in treatment. EPID images with 512 x 384 pixels (pixel size: 0.56 mm) were acquired at a sampling rate of 0.5 frame s(-1) by using energies of 4, 6 or 10 MV on linear accelerators. We applied our proposed method to EPID cine images (226 frames) of 12 clinical cases (ages: 51-83, mean: 72) with a non-small cell lung cancer. As a result, the average location error between tumor points obtained by our method and the manual method was 1.47 +/- 0.60 mm. This preliminary study suggests that our method based on the tumor template matching technique might be feasible for tracking the location of a lung tumor without implanted markers in SBRT. PMID- 19131669 TI - Quantitative three-dimensional elasticity imaging from quasi-static deformation: a phantom study. AB - We present a methodology to image and quantify the shear elastic modulus of three dimensional (3D) breast tissue volumes held in compression under conditions similar to those of a clinical mammography system. Tissue phantoms are made to mimic the ultrasonic and mechanical properties of breast tissue. Stiff lesions are created in these phantoms with size and modulus contrast values, relative to the background, that are within the range of values of clinical interest. A two dimensional ultrasound system, scanned elevationally, is used to acquire 3D images of these phantoms as they are held in compression. From two 3D ultrasound images, acquired at different compressed states, a three-dimensional displacement vector field is measured. The measured displacement field is then used to solve an inverse problem, assuming the phantom material to be an incompressible, linear elastic solid, to recover the shear modulus distribution within the imaged volume. The reconstructed values are then compared to values measured independently by direct mechanical testing. PMID- 19131670 TI - A theoretical model for respiratory motion artifacts in free-breathing CT scans. AB - Successful radiotherapy treatment depends heavily upon the accuracy of patient geometry captured during treatment simulation using computed tomography (CT) scans. Radiotherapy patients are often scanned under free breathing, and respiratory motion can cause severe artifacts in CT scans, including shortening, elongation or splitting of the shapes and shifting of the midpoint positions of the tumor and organs. This paper presents a theoretical model that explains the source of motion artifacts and the relationship between motion artifacts and the motion parameters of the scanner, treatment couch and tumor/organ. It is shown that an understanding of the relationship between the translational table velocity and the maximum tumor/organ velocity might enable one to mitigate certain types of motion artifacts. We show that splitting artifacts can be eliminated if the scanning speed is above the maximum tumor/organ velocity. Slow scanning speeds are shown to be useful for obtaining accurate internal target volumes (ITVs), and fast scanning speeds are shown to be useful for obtaining accurate tumor/organ shapes. In both cases, an upper bound on the maximum possible error is calculated as a function of the scanning speed. A set of special scanning speeds which allow for an accurate representation of tumor/organ length along the craniocaudal direction is obtained, and a relationship between the maximum displacement of a tumor/organ image's midpoint position and the magnitude of its length distortion is derived. PMID- 19131671 TI - Mapping metals in Parkinson's and normal brain using rapid-scanning x-ray fluorescence. AB - Rapid-scanning x-ray fluorescence (RS-XRF) is a synchrotron technology that maps multiple metals in tissues by employing unique hardware and software to increase scanning speed. RS-XRF was validated by mapping and quantifying iron, zinc and copper in brain slices from Parkinson's disease (PD) and unaffected subjects. Regions and structures in the brain were readily identified by their metal complement and each metal had a unique distribution. Many zinc-rich brain regions were low in iron and vice versa. The location and amount of iron in brain regions known to be affected in PD agreed with analyses using other methods. Sample preparation is simple and standard formalin-fixed autopsy slices are suitable. RS XRF can simultaneously and non-destructively map and quantify multiple metals and holds great promise to reveal metal pathologies associated with PD and other neurodegenerative diseases as well as diseases of metal metabolism. PMID- 19131672 TI - Motion correction of PET brain images through deconvolution: I. Theoretical development and analysis in software simulations. AB - Image quality is significantly degraded even by small amounts of patient motion in very high-resolution PET scanners. Existing correction methods that use known patient motion obtained from tracking devices either require multi-frame acquisitions, detailed knowledge of the scanner, or specialized reconstruction algorithms. A deconvolution algorithm has been developed that alleviates these drawbacks by using the reconstructed image to estimate the original non-blurred image using maximum likelihood estimation maximization (MLEM) techniques. A high resolution digital phantom was created by shape-based interpolation of the digital Hoffman brain phantom. Three different sets of 20 movements were applied to the phantom. For each frame of the motion, sinograms with attenuation and three levels of noise were simulated and then reconstructed using filtered backprojection. The average of the 20 frames was considered the motion blurred image, which was restored with the deconvolution algorithm. After correction, contrast increased from a mean of 2.0, 1.8 and 1.4 in the motion blurred images, for the three increasing amounts of movement, to a mean of 2.5, 2.4 and 2.2. Mean error was reduced by an average of 55% with motion correction. In conclusion, deconvolution can be used for correction of motion blur when subject motion is known. PMID- 19131673 TI - Prompt gamma-ray emission from biological tissues during proton irradiation: a preliminary study. AB - In this paper, we present the results of a preliminary study of secondary 'prompt' gamma-ray emission produced by proton-nuclear interactions within tissue during proton radiotherapy. Monte Carlo simulations were performed for mono energetic proton beams, ranging from 2.5 MeV to 250 MeV, irradiating elemental and tissue targets. Calculations of the emission spectra from different biological tissues and their elemental components were made. Also, prompt gamma rays emitted during delivery of a clinical proton spread-out Bragg peak (SOBP) in a homogeneous water phantom and a water phantom containing heterogeneous tissue inserts were calculated to study the correlation between prompt gamma-ray production and proton dose delivery. The results show that the prompt gamma-ray spectra differ significantly for each type of tissue studied. The relative intensity of the characteristic gamma rays emitted from a given tissue was shown to be proportional to the concentration of each element in that tissue. A strong correlation was found between the delivered SOBP dose distribution and the characteristic prompt gamma-ray production. Based on these results, we discuss the potential use of prompt gamma-ray emission as a method to verify the accuracy and efficacy of doses delivered with proton radiotherapy. PMID- 19131674 TI - Laser-Doppler spectrum decomposition applied for the estimation of speed distribution of particles moving in a multiple scattering medium. AB - Recently, a method for the estimation of speed distribution of particles moving in an optically turbid medium has been proposed. The method allows potentially absolute measurement of speed of the particles and can be applied in laser Doppler perfusion measurements. However, the decomposition technique was limited to short source-detector separations for which the assumption that one photon is Doppler scattered not more than once is fulfilled. In the present paper we show a generalized decomposition technique in which photons can be scattered more than once. We show the theoretical background for decomposition in such a case. We apply a decomposition method for the analysis of laser-Doppler spectra obtained by Monte Carlo simulations. This analysis allows showing noise limits in which the technique can be effectively applied in analysis of measured spectra. We propose an approximated scattering model based on the assumption that for one photon consecutive Doppler scattering events occur on particles moving with the same speed, and we show that this approximation does not influence significantly the uncertainty of the resulting speed distribution. The proposed decomposition procedure is validated in measurements on a physical flow model. The decomposition procedure is also validated by analysis of spectra measured on a physical phantom using laser-Doppler flow meter (Oxford Optronix, UK). A diluted solution of milk was pumped through a tube fixed in an optically turbid material with speed varying from 0 mm s(-1) to 4 mm s(-1). We observed a linear relation between actual speed of milk solution and speed estimated from results of spectra decomposition. PMID- 19131675 TI - Evaluating the cancer detection and grading potential of prostatic-zinc imaging: a simulation study. AB - The present work deals with the analysis of prostatic-zinc-concentration images. The goal is to evaluate potential clinically relevant information that can be extracted from such images. In the absence of experimental images, synthetic ones are produced from clinically measured zinc-concentration distributions in certified benign and cancerous tissue samples, classified by the lesion grade. We describe the method for producing the images and model the effect of counting statistics noise. We present in detail the image analysis, which is based on a combination of standard image processing and segmentation tools, optimized for this particular application. The information on lowest zinc value obtained from the image analysis is translated to clinical data such as tumour presence, location, size and grade. Their confidence is evaluated with the help of standard statistical tools such as receiver operating characteristic analysis. The present work predicts a potential for detecting small prostate-cancer lesions, of grade (4+3) and above, with very good specificity and sensitivity. The present analysis further provides data on the pixel size and image counting statistics requested from the trans-rectal probe that will record in vivo prostatic-zinc maps in patients. PMID- 19131676 TI - Local complex permittivity measurements of porcine skin tissue in the frequency range from 1 GHz to 15 GHz by evanescent microscopy. AB - The near-field evanescent microwave microscope is based on a coaxial transmission line resonator with a silver plated tungsten tip protruding through an end-wall aperture. The sensor is used to measure the local dielectric properties of porcine skin in the frequency range from 1 GHz to 15 GHz. The dielectric property of the skin within the near field of the tip frustrates the electric field and measurably changes the transmission line's resonant frequency and quality factor (Q). The shift of the resonator's frequency and Q is measured as a function of tip-sample separation, and a quantitative relationship between the real and imaginary parts of the local dielectric constant using the method of images is established. The associated changes in quality factor image scans of subsurface tissue structure and dielectric properties of skin surface lesions are presented. PMID- 19131677 TI - Differences in hip kinematics, muscle strength, and muscle activation between subjects with and without patellofemoral pain. AB - STUDY DESIGN: Controlled laboratory study using a cross-sectional design. OBJECTIVES: To determine whether females with patellofemoral pain (PFP) demonstrate differences in hip kinematics, hip muscle strength, and hip muscle activation patterns when compared to pain-free controls. BACKGROUND: It has been proposed that abnormal hip kinematics may contribute to the development of PFP. However, research linking hip function to PFP remains limited. METHODS AND MEASURES: Twenty-one females with PFP and 20 pain-free controls participated in this study. Hip kinematics and activity level of hip musculature were obtained during running, a drop jump, and a step-down maneuver. Isometric hip muscle torque production was quantified using a multimodal dynamometer. Group differences were assessed across tasks using mixed-design 2-way analyses of variance and independent t tests. RESULTS: When averaged across all 3 activities, females with PFP demonstrated greater peak hip internal rotation compared to the control group (mean +/- SD, 7.6 degrees +/- 7.0 degrees versus 1.2 degrees +/- 3.8 degrees; P<.05). The individuals in the PFP group also exhibited diminished hip torque production compared to the control group (14% less hip abductor strength and 17% less hip extensor strength). Significantly greater gluteus maximus recruitment was observed for individuals in the PFP group during running and the step-down task. CONCLUSION: The increased peak hip internal rotation motion observed for females in the PFP group was accompanied by decreased hip muscle strength. The increased activation of the gluteus maximus in individuals with PFP suggests that these subjects were attempting to recruit a weakened muscle, perhaps in an effort to stabilize the hip joint. Our results support the proposed link between abnormal hip function and PFP. PMID- 19131678 TI - The change in the diffusion of water in normal and degenerative lumbar intervertebral discs following joint mobilization compared to prone lying. AB - STUDY DESIGN: Prospective, repeated measures obtained under treatment and control conditions. OBJECTIVES: The purposes of this study were to provide preliminary evidence regarding the immediate change in the diffusion of water in the nuclear region of normal and degenerative lumbar intervertebral discs (IVDs) following a single session of lumbar joint mobilization, and to compare these findings to the immediate change in the diffusion of water following a 10-minute session of prone lying. BACKGROUND: There is conflicting evidence regarding the effectiveness and efficacy of lumbar joint mobilization. Increased knowledge of the physiologic effects of lumbar joint mobilization can lead to refinement of its clinical application. METHODS AND MEASURES: A total of 24 people (15 males and 9 females), ranging in age from 22 to 58 years, participated in this study. All subjects had a history of activity-limiting low back pain. Diffusion-weighted magnetic resonance images (DW-MRIs) were obtained immediately before and after a 10-minute session of lumbar joint mobilization. At least 1 month later, a second session was performed in which DW-MRIs were obtained immediately before and after a 10 minute session of prone lying. RESULTS: Following lumbar joint mobilization, a significant increase (P = .002) in the mean values for diffusion of water was observed within degenerative IVDs at L5-S1 (22.2% increase; effect size, 0.97). Degenerative IVDs at L1-2 to L4-5 and normal IVDs at L1-2 to L5-S1 did not demonstrate a change in diffusion following joint mobilization. Prone lying was not associated with a change in diffusion for normal or degenerative IVDs. CONCLUSIONS: The stimulus provided by lumbar joint mobilization may influence the diffusion of water in degenerative IVDs at L5-S1; however, these are preliminary findings and the relationship of these findings to pain and function needs further investigation. PMID- 19131680 TI - Tarsometatarsal joint injury in a patient seen in a direct-access physical therapy setting. PMID- 19131686 TI - Identification of psychrophile Shewanella sp. KMG427 as an eicosapentaenoic acid producer. AB - An isolate from holothurians was identified as an eicosapentaenoic acid (EPA) producing bacterium KMG427, which is characterized by EPA synthesis efficiency, by thin layer and gas chromatographic analyses. The EPA production was maximized to more than 10% of the total fatty acids by incubation at 4o degrees C after cell proliferation at 20 degrees C. The isolated bacterium was categorized as Gramnegative, rod-shaped, aerobic, and motile with a single polar flagellum. According to phylogenetic analysis based on morphological and physiological specificities as an EPA-producing bacterium, the isolate KMG427 was found to belong to the genus Shewanella. The 16S rDNA of KMG427 was revealed to have 100% of sequence identity to that of S. hanedai CIP 103207T. Therefore, the isolate might be classified and identified as Shewanella sp. KMG427. PMID- 19131688 TI - Involvement of beta-lactamase inhibitory protein, BLIP-II, in morphological differentiation of Streptomyces exfoliatus SMF19. AB - The beta-lactamase inhibitory protein, BLIP-II, found in the culture supernatant of Streptomyces exfoliatus SMF19, shows no discernible sequence identity with other beta-lactamase inhibitory proteins identified in Streptomyces spp. A null mutant of the gene encoding BLIP-II (bliB::hygr) showed a bald appearance on solid media. Although BLIP-II was initially isolated from the supernatant of submerged cultures, sites of BLIP-II accumulation were seen in the cell envelope. The mutation of bliB was also associated with changes in the formation of septa and condensation of the chromosomal DNA associated with sporulation. The bliB mutant exhibited infrequent septa, showing dispersed chromosomal DNA throughout the mycelium, whereas the condensed chromosomes of the wild-type were separated by regularly spaced septa giving the appearance of a string of beads. Therefore, on the basis of these results, it is suggested that BLIP-II is a regulator of morphological differentiation in S. exfoliatus SMF19. PMID- 19131687 TI - Culture-independent analysis of microbial succession during composting of Swine slurry and mushroom cultural wastes. AB - Bacterial diversity and the composition of individual communities during the composting process of swine and mushroom cultural wastes in a field-scale composter (Hazaka system) were examined using a PCR-based approach. The composting process was divided into six stages based on recorded temperature changes. Phylogenetic analysis of eighty 16S rRNA sequences from uncultured composting bacterial groups revealed the presence of representatives from three divisions, including plant pathogenic bacteria, high-molecule-degrading bacteria and spore-forming bacteria. The plant pathogen A. tumefaciens gradually decreased in abundance during the composting process and eventually disappeared during the thermophilic and cooling stage. A bacterium homologous to Bacillus humi first appeared at the early thermophilic stage and was established at the intermediate thermophilic, post-thermophilic, and cooling stages. It was not possible to isolate the B. humi during any of the stages using general culture techniques. PMID- 19131689 TI - Influence of deletions in the apoemulsan gene cluster on A. venetianus RAG-1 polysaccharide biosynthesis. AB - Apoemulsan is a biopolymer with potent emulsification activity, produced by Acinetobacter venetianus RAG-1 (RAG-1). The wee gene cluster is responsible for apoemulsan biosynthesis. The analysis of (i) a putative polysaccharide copolymerase mutant (wzc), (ii) a putative polymerase mutant (wzy), and (iii) an apoemulsan-deficient variant (2) indicated that the wee gene cluster controls the synthesis of two polysaccharides: high molecular weight (HMW) and low molecular weight (LMW). LMW polysaccharide of wee origin was present in LPS isolated from RAG-1 cells, suggesting a link to the Lipid A-core of LPS molecules. SDSPAGE analysis indicated that apoemulsan is copurified with LPS polysaccharide, with implications in the emulsification activity of RAG-1 polymer. PMID- 19131690 TI - Proteome analysis for 3T3-L1 adipocyte differentiation. AB - Adipose tissue is an important endocrine organ involved in the control of whole body energy homeostasis and insulin sensitivity. Considering the increased incidence of obesity and obesity-related disorders, including diabetes, it is important to understand thoroughly the process of adipocyte differentiation and its control. Therefore, we performed a differential proteome mapping strategy using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to identify intracellular proteins that are differentially expressed during adipose conversion of 3T3-L1 pre-adipocytes in response to an adipogenic cocktail. In the current study, we identified 46 differentially expressed proteins, 6 of which have not been addressed previously in 3T3-L1 cell differentiation. Notably, we found that phosphoribosyl pyrophosphate synthetase (PRPS), a regulator of cell proliferation, was preferentially expressed in pre adipocytes than in fully differentiated adipocytes. In conclusion, our results provide valuable information for further understanding of the adipogenic process. PMID- 19131691 TI - Statistical optimization of medium components for the production of prodigiosin by Hahella chejuensis KCTC 2396. AB - Prodigiosin is a natural red pigment with algicidal activity against Cochlodinium polykrikoides, a major harmful redtide microalga. To increase the yield of prodigiosin production by Hahella chejuensis KCTC 2396, significant medium components were determined using a two-level Plackett- Burman statistical design technique. Among 12 components included in basal medium, NaHCO3, Na2SiO3, NH4NO3, Na2SO4, and CaCl2 were determined to be important for prodigiosin production. The medium formulation was finally optimized using a Box-Behnken design as follows: 1% sucrose; 0.4% peptone; 0.1% yeast extract; and (g/l): NaCl, 20.0; Na2SO4, 9.0; CaCl2, 1.71; KCl, 0.4; and (mg/l): H3BO3, 10.0; KBr, 50.0; NaF, 2.0; NaHCO3, 45.0; Na2SiO3, 4.5; NH4NO3, 4.5. The predicted maximum yield of prodigiosin in the optimized medium was 1.198 g/l by the Box-Behnken design, whereas the practical production was 1.495 g/l, which was three times higher than the basal medium (0.492 g/l). PMID- 19131692 TI - Isolation and physiological characterization of Bacillus clausii SKAL-16 isolated from wastewater. AB - An alkaliphilic bacterium, Bacillus clausii SKAL-16, was isolated from soil that had been contaminated with vegetable oil. The optimal pH and general pH range for bacterial growth was 8, and 7 to 10, respectively. The bacterium could grow on tributyrin and glycerol, but could not grow on acetate and butyrate. The SKAL-16 strain excreted butyric acid during growth on tributyrin, and selectively ingested glycerol during growth on a mixture of butyric acid and glycerol. The SKAL-16 generated intracellular lipase, but did not produce esterase and extracellular lipase. The DNA fragment amplified with the chromosomal DNA of SKAL 16 and primers designed on the basis of the esterase-coding gene of Bacillus clausii KSM-K16 was not identical with the esterase-coding gene contained in the GenBank database. Pyruvate dehydrogenase, isocitrate dehydrogenase, and malate dehydrogenase activities were detected in the cellfree extract (crude enzyme). PMID- 19131693 TI - Laboratory and field evaluations of entomopathogenic Lecanicillium attenuatum CNU 23 for control of green peach aphid (Myzus persicae). AB - An entomopathogenic fungus was isolated from an infected aphid. The isolate conformed most closely to Lecanicillium attenuatum CBS 402.78 (AJ292434) based on the internal transcribed spacer (ITS) region of its 18S rDNA, and thus was designated L. attenuatum CNU-23. Laboratory and field evaluations of CNU-23 blastospores were carried out for the control of green peach aphids. The laboratory evaluations of CNU-23 revealed an aphid mortality of about 80% with an estimated LT50 of 3.72 days after the application of CNU-23 at 1x106 blastospores/ ml. Meanwhile, the field evaluations of CNU-23 performed on greenhouse pepper plants during the rainy season showed an aphid mortality ranging from 72% to 97%. Significant sporulation was observed in the aphids treated with CNU-23. Therefore, the results suggest that L. attenuatum CNU-23 can be used as a biocontrol agent for green peach aphids on greenhouse pepper plants. PMID- 19131694 TI - Statistical optimization of the growth factors for Chaetoceros neogracile using fractional factorial design and central composite design. AB - Statistical experimental designs; involving (i) a fractional factorial design (FFD) and (ii) a central composite design (CCD) were applied to optimize the culture medium constituents for production of a unique antifreeze protein by the Antartic microalgae Chaetoceros neogracile. The results of the FFD suggested that NaCl, KCl, MgCl2, and Na2SiO3 were significant variables that highly influenced the growth rate and biomass production. The optimum culture medium for the production of an antifreeze protein from C. neogracile was found to be Kalleampersandrsquor;s artificial seawater, pH of 7.0ampersandplusmn;0.5, consisting of 28.566 g/l of NaCl, 3.887 g/l of MgCl2, 1.787 g/l of MgSO4, 1.308 g/l of CaSO4, 0.832 g/l of K2SO4, 0.124 g/l of CaCO3, 0.103 g/l of KBr, 0.0288 g/l of SrSO4, and 0.0282 g/l of H3BO3. The antifreeze activity significantly increased after cells were treated with cold shock (at -5oC) for 14 h. To the best of our knowledge, this is the first report demonstrating an antifreeze-like protein of C. neogracile. PMID- 19131696 TI - Improvement of amidase production by a newly isolated Delftia tsuruhatensis ZJB 05174 through optimization of culture medium. AB - The R-amidase production by a newly isolated strain of Delftia tsuruhatensis ZJB 05174 was optimized in this paper. Effects of factors such as carbon sources, nitrogen sources, and inducers on amidase production were investigated. The medium composition was optimized using central composite designs and response surface analysis. The optimal medium components for enhanced amidase production were found to be as follows: glucose, 8.23 g/l; yeast extract, 11.59 g/l; 2,2 (R,S)-dimethylcyclopropane carboxamide, 1.76 g/l; NaCl, 1 g/l; KH2PO4, 1 g/l; and K2HPO4, 1 g/l. A maximum enzyme production of 528.21 U/l was obtained under the optimized conditions, which was 4.7 times higher than that obtained under initial conditions. PMID- 19131695 TI - Optimization of the process for biodiesel production using a mixture of immobilized Rhizopus oryzae and Candida rugosa lipases. AB - In this study, the enzymatic process for biodiesel production was optimized using a mixture of immobilized Rhizopus oryzae and Candida rugosa lipases. The optimal temperature and agitation speed for biodiesel production were 45oC and 300 rpm, respectively. The optimal ratio of R. oryzae and C. rugosa lipases in the mixture was 3:1 (w:w). When 3mmol of methanol was the initial reaction medium and 3mmol of methanol was added every 1.5 h during biodiesel production, biodiesel conversion was over 98% at 4 h. In addition, when the immobilized lipase mixture was reused, biodiesel conversion exceeded 80% after 5 reuses. PMID- 19131697 TI - High-level secretory expression of human procarboxypeptidase B by Fed-Batch cultivation of Pichia pastoris and its partial characterization. AB - The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, pPIC9alpha/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOX1 promoter and connected to the downstream of the mating factor alpha-1 (MFalpha1) signal sequence. The plasmid was linearized by digestion with SacI, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fedbatch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The Km and kcat values of recombinant human CPB enzyme for hippuryl-L-Arg as a substrate were estimated to be 0.16mM and 11.93 sec-1, respectively. PMID- 19131698 TI - Enhanced sialylation of recombinant erythropoietin in CHO cells by human glycosyltransferase expression. AB - Sialylation, the attachment of sialic acid residues to a protein, can affect the biological activity and in vivo circulatory half-life of glycoproteins. Human alpha2,3- sialyltransferase (alpha2,3-ST) and beta1,4-galactosyltransferase (beta1,4-GT) are responsible for terminal sialylation and galactosylation, respectively. Enhanced sialylation of human erythropoietin (EPO) by the expression of alpha2,3-ST and beta1,4-GT was achieved using recombinant Chinese hamster ovary (CHO) cells (EC1). The sialic acid content and sialylation of N glycans were evaluated by HPLC. When alpha2,3-ST was expressed in CHO cells (EC1 ST2), the sialic acid content (moles of sialic acid/mole of EPO) increased from 6.7 to 7.5. In addition, the amount of trisialylated glycans increased from 17.3% to 26.1%. When alpha2,3-ST and beta1,4-GT were coexpressed in CHO cells (EC1 GTST15), the degree of sialylation was greater than that in EC1-ST2 cells. In the case of EC1-GTST15 cells, the sialic acid content increased to 8.2 and the proportion of trisialylated glycans was markedly increased from 17.3% to 35.5%. Interestingly, the amount of asialoglycans decreased only in the case of GTST15 cells (21.4% to 14.2%). These results show that coexpression of alpha2,3- ST and beta1,4-GT is more effective than the expression of alpha2,3-ST alone. Coexpression of alpha2,3-ST and beta1,4-GT did not affect CHO cell growth and metabolism or EPO production. Thus, coexpression of alpha2,3-ST and beta1,4-GT may be beneficial for producing therapeutic glycoproteins with enhanced sialylation in CHO cells. PMID- 19131699 TI - Construction of recombinant Lactobacillus casei strains using splicing by overlap extension. AB - Recombinant Lactobacillus strains have been constructed using gene splicing by overlap extension (SOE). Primers were designed of which one end of an amplified product contained complementary sequences for an end of other amplified fragment. For efficient matching, we used an asymmetric PCR step that was effective at generating an excess of strands that would anneal in the final PCR. CP12, a recombinant fragment consisting of the integrase gene and attachment site of the bacteriophage A2, was constructed and inserted into the genome of Lactobacillus casei ATCC 393, yielding Lb. casei ATCC 393::XCP12. Another recombinant Lb. casei strain was constructed, where the egfp gene was a part of the construction. The EGFP produced from Lb. casei ATCC 393::XCEGFP14 was detected by Western blot hybridization. This simple and widely applicable approach has significant advantages over standard recombinant DNA techniques for Lactobacillus species. PMID- 19131700 TI - Characterizations of denitrifying polyphosphate-accumulating bacterium Paracoccus sp. Strain YKP-9. AB - A denitrifying polyphosphate-accumulating bacterium (YKP-9) was isolated from activated sludge of a 5-stage biological nutrient removal process with step feed system. This organism was a Gram-negative, coccus-shaped, facultative aerobic chemoorganotroph. It had a respiratory type of metabolism with oxygen, nitrate, and nitrite as terminal electron acceptors. The 16S rRNA gene sequence of strain YKP-9 was most similar to the 16S rRNA gene sequence of Paracoccus sp. OL18 (AY312056) (similarity level, 97%). Denitrifying polyphosphate accumulation by strain YKP-9 was examined under anaerobicanoxic and anaerobic-oxic batch conditions. It was able to use external carbon sources for polyhydroxyalkanoates (PHA) synthesis and to release phosphate under anaerobic condition. It accumulated polyphosphate and grew a little on energy provided by external carbon sources under anoxic condition, but did neither accumulate polyphosphate nor grow in the absence of external carbon sources under anoxic condition. Cells with intracellular PHA cannot accumulate polyphosphate in the absence of external carbon sources under anoxic condition. Under oxic condition, it grew but could not accumulate polyphosphate with external carbon sources. Based on the results from this study, strain YKP-9 is a new-type denitrifying polyphosphate accumulating bacterium that accumulates polyphosphate only under anoxic condition, with nitrate and nitrite as the electron acceptors in the presence of external carbon sources. PMID- 19131701 TI - Bioremediation potential of a tropical soil contaminated with a mixture of crude oil and production water. AB - A typical tropical soil from the northeast of Brazil, where an important terrestrial oil field is located, was accidentally contaminated with a mixture of oil and saline production water. To study the bioremediation potential in this area, molecular methods based on PCR-DGGE were used to determine the diversity of the bacterial communities in bulk and in contaminated soils. Bacterial fingerprints revealed that the bacterial communities were affected by the presence of the mixture of oil and production water, and different profiles were observed when the contaminated soils were compared with the control. Halotolerant strains capable of degrading crude oil were also isolated from enrichment cultures obtained from the contaminated soil samples. Twenty-two strains showing these features were characterized genetically by amplified ribosomal DNA restriction analysis (ARDRA) and phenotypically by their colonial morphology and tolerance to high NaCl concentrations. Fifteen ARDRA groups were formed. Selected strains were analyzed by 16S rDNA sequencing, and Actinobacteria was identified as the main group found. Strains were also tested for their growth capability in the presence of different oil derivatives (hexane, dodecane, hexadecane, diesel, gasoline, toluene, naphthalene, o-xylene, and p-xylene) and different degradation profiles were observed. PCR products were obtained from 12 of the 15 ARDRA representatives when they were screened for the presence of the alkane hydroxylase gene (alkB). Members of the genera Rhodococcus and Gordonia were identified as predominant in the soil studied. These genera are usually implicated in oil degradation processes and, as such, the potential for bioremediation in this area can be considered as feasible. PMID- 19131702 TI - Lactobacillus casei secreting alpha-MSH induces the therapeutic effect on DSS induced acute colitis in Balb/c Mice. AB - The neuropeptide alpha-melanocyte-stimulating hormone (alpha- MSH) has anti inflammatory property by downregulating the expressions of proinflammatory cytokines. Because alpha-MSH elicits the anti-inflammatory effect in various inflammatory disease models, we examined the therapeutic effect of oral administration of recombinant Lactobacillus casei, which secretes alpha-MSH (L. casei-alpha-MSH), on dextran sulfate sodium (DSS)-induced colitis in Balb/c mice. Thus, we constructed the alpha-MSH-secreting Lactobacillus casei by the basic plasmid, pLUAT-ss, which was composed of a PldhUTLS promoter and alpha-amylase signal sequence from Streptococcus bovis strain. Acute colitis was induced by oral administration of 5% DSS in drinking water for 7 days. To investigate the effect of L. casei-alpha-MSH on the colitis, L. casei or L. casei-alpha-MSH was orally administered for 7 days and their effects on body weight, mortality rate, cytokine production, and tissue myeloperoxidase (MPO) activity were observed. Administration of L. casei-alpha-MSH reduced the symptom of acute colitis as assessed by body weight loss (DSS alone: 14.45+/-0. 2 g; L. casei-alpha- MSH: 18.2+/-0.12 g), colitis score (DSS alone: 3.6+/-0.4; L. casei-alpha-MSH: 1.4+/ 0.6), MPO activity (DSS alone: 42.7+/-4.5 U/g; L. casei-alpha-MSH: 10.25+/-0.5 U/g), survival rate, and histological damage compared with the DSS alone mice. L. casei-alpha-MSH-administered entire colon showed reduced in vitro production of proinflammatory cytokines and NF-kappaB activation. The alpha-MSH-secreting recombinant L. casei showed significant anti-inflammatory effects in the murine model of acute colitis and suggests a potential therapeutic role for this agent in clinical inflammatory bowel diseases. PMID- 19131703 TI - Surfactin blocks NO production in lipopolysaccharide-activated macrophages by inhibiting NF-kappaB activation. AB - Surfactin is a natural biosurfactant derived from Bacillus subtilis and has various biological activities such as anticancer, antiplatelet, and anti inflammatory effects. In this study, the inhibitory mechanism of surfactin in NO production from macrophages was examined. Surfactin downregulated LPSinduced NO production in RAW264.7 cells and primary macrophages with IC50 values of 31.6 and 22.4 microM, respectively. Immunoblotting analysis showed that surfactin strongly blocked the phosphorylation of IKK and IkBa and the nuclear translocation of NF kappaB (p65). Therefore, these data suggest that surfactin may act as a bacterium derived antiinflammatory agent with anti-NF-kappaB activity. PMID- 19131704 TI - Effects of astaxanthin on the production of NO and the expression of COX-2 and iNOS in LPS-stimulated BV2 microglial cells. AB - Astaxanthin has shown antioxidant, antitumor, and antiinflammatory activities; however, its molecular action and mechanism in the nervous system have yet to be elucidated. We examined the in vitro effects of astaxanthin on the production of nitric oxide (NO), as well as the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Astaxanthin inhibited the expression or formation of nitric oxide (NO), iNOS and COX-2 in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Astaxanthin also suppressed the protein levels of iNOS and COX-2 in LPS stimulated BV2 microglial cells. These results suggest that astaxanthin, probably due to its antioxidant activity, inhibits the production of inflammatory mediators by blocking iNOS and COX-2 activation or by the suppression of iNOS and COX-2 degradation. PMID- 19131705 TI - Induction of apoptosis by aqueous extract of Cordyceps militaris through activation of caspases and inactivation of Akt in human breast cancer MDA-MB-231 Cells. AB - Cordyceps militaris is well known as a traditional medicinal mushroom and has been shown to exhibit immunostimulatory and anticancer activities. In this study, we investigated the apoptosis induced by an aqueous extract of C. militaris (AECM) via the activation of caspases and altered mitochondrial membrane permeability in human breast cancer MDA-MB-231 cells. Exposure to AECM induced apoptosis, as demonstrated by a quantitative analysis of nuclear morphological change and a flow cytometric analysis. AECM increased hyperpolarization of mitochondrial membrane potential and promoted the activation of caspases. Both the cytotoxic effect and apoptotic characteristics induced by AECM treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role of caspase-3 in the observed cytotoxic effect. AECM-induced apoptosis was associated with the inhibition of Akt activation in a time dependent manner, and pretreatment with LY294002, a PI3K/Akt inhibitor, significantly increased AECM-induced apoptosis. The results indicated that AECM induced apoptosis may relate to the activation of caspase-3 and mitochondria dysfunctions that correlate with the inactivation of Akt. PMID- 19131706 TI - Differential regulation of the PIKK kinase family by Tel2. PMID- 19131707 TI - Is health care a commodity? PMID- 19131708 TI - The Edwin Smith papyrus (ca. 16th century BC). PMID- 19131709 TI - Correlation between complication rate and patient satisfaction in abdominoplasty. AB - Abdominoplasty has become one of the most frequently performed procedures for improving body contour. Numerous reports exist focusing on postoperative complications with even more studies suggesting technical modifications to decrease their occurrence. However, the impact of complications on patient satisfaction has not been sufficiently addressed to date. A retrospective analysis was performed investigating the occurrence of complications following abdominoplasty and their effect on patient satisfaction. Patient satisfaction was assessed by means of the client satisfaction questionnaire-8. From June 1994 to April 2004 a total of 139 patients underwent an abdominoplasty. Minor and major complications were encountered in 40 (28.8%) and 16 (11.5%) patients, respectively. The median client satisfaction questionnaire-8 score among those patients with and without complications was 29. Thus, it seems that postoperative complications after abdominoplasty do not negatively affect patient satisfaction. PMID- 19131710 TI - Cosmetic surgery reality TV viewership: relations with cosmetic surgery attitudes, body image, and disordered eating. AB - BACKGROUND: According to the American Society of Plastic Surgeons (2007), the number of cosmetic procedures has increased to over 10 million in 2006, representing a 48% increase from 2000. This increase in cosmetic surgery prevalence is paralleled by a surge in reality cosmetic surgery television programming. METHODS: The current study examined the relationships among cosmetic surgery reality TV viewership, cosmetic surgery attitudes, body image, and disordered eating in a sample of 2057 college women. RESULTS: Viewership of reality cosmetic surgery shows was significantly related to more favorable cosmetic surgery attitudes, perceived pressure to have cosmetic surgery, past attainment of a cosmetic procedure, a decreased fear of surgery, as well as overall body dissatisfaction, media internalization, and disordered eating. CONCLUSIONS: Although the current study is correlational, it provides a framework for future hypothesis testing and elucidates the link between contemporary media influences, body dissatisfaction, disordered eating, and cosmetic surgery attitudes. Additionally, the findings indicate that surgeons may want to assess the relevance of cosmetic surgery reality TV viewership for patients' attitudes towards and expectations about cosmetic surgery. PMID- 19131711 TI - Multisystem organ failure after large volume injection of castor oil. AB - We report a case of multisystem organ failure after large volume subcutaneous injection of castor oil for cosmetic enhancement. An unlicensed practitioner injected 500 mL of castor oil bilaterally to the hips and buttocks of a 28-year old male to female transsexual. Immediate local pain and erythema were followed by abdominal and chest pain, emesis, headache, hematuria, jaundice, and tinnitus. She presented to an emergency department 12 hours postinjection. Persistently hemolyzed blood samples complicated preliminary laboratory analysis. She rapidly deteriorated despite treatment and developed fever, tachycardia, hemolysis, thrombocytopenia, hepatitis, respiratory distress, and anuric renal failure. An infectious diseases evaluation was negative. After intensive supportive care, including mechanical ventilation and hemodialysis, she was discharged 11 days later, requiring dialysis for an additional 1.5 months. Castor oil absorption was inferred from recovery of the Ricinus communis biomarker, ricinine, in the patient's urine (41 ng/mL). Clinicians should anticipate multiple complications after unapproved methods of cosmetic enhancement. PMID- 19131712 TI - Efficacy of pocket irrigation with bupivacaine and ketorolac in breast augmentation: a randomized controlled trial. AB - Breast augmentation is the most common cosmetic surgery procedure performed in the United States. The optimal approach to postoperative pain management in these patients, however, has yet to be determined. The objective of this study was to investigate the efficacy of pocket irrigation with bupivacaine and ketorolac in reducing pain, narcotic use, and methocarbamol use following subpectoral breast augmentation. A prospective, double-blinded, randomized trial was performed. Fifty primary breast augmentation patients were randomized to 1 of 2 treatment groups: 1) pocket irrigation with bupivacaine and ketorolac, and 2) no pocket irrigation. Patients who received pocket irrigation had significantly lower Visual Analog Pain Scores in the early postoperative period (1 and 6 hours postoperatively). There was no difference in early postoperative narcotic use in patients who received pocket irrigation and those who did not. Patients who received pocket irrigation, however, reported higher narcotic use at 1-3 days following surgery. There was no difference in postoperative methocarbamol use between patient groups. This study suggests that pocket irrigation with bupivacaine and ketorolac can significantly decrease patient-reported pain in the early postoperative period following subpectoral augmentation. It also demonstrates, however, that there is an increased requirement for narcotics between 1-3 days after surgery. These findings suggest that intraoperative administration of analgesics into the implant pocket may thus facilitate an early postoperative recovery; yet, patients should be advised that they might require more pain medication at home for the first few days. PMID- 19131713 TI - Huge cystic lymphangioma involving the entire breast. AB - We report a case of a 31-year-old female who required mastectomy with the diagnosis of cystic lymphangioma of the entire left breast. We tried operative management that is not mastectomy since it was not malignant, but finally mastectomy was needed. In reduction mammaplasty patients, plastic surgeons should consider cystic lymphangioma as a differential diagnosis of unilateral large breasts. PMID- 19131714 TI - A prognostic model for the risk of development of upper extremity compartment syndrome in the setting of brachial artery injury. AB - A potentially devastating sequela of brachial artery injury in the setting of upper extremity trauma is the development of compartment syndrome (CS). We performed a retrospective review of 139 trauma patients with brachial artery injury from 1985-2001. Objective characteristics of each case were extracted and analyzed using multivariate logistic regression. Three variables were found to be significant in the final model: estimated intraoperative blood loss as a continuous variable, and presence of a multiple arterial injury and presence of an open fracture as categorical variables. Odds ratio were 1.12, 5.79, and 2.68, respectively. We used these variables to create a summative score for the development of CS with weights assigned proportional to the adjusted odds ratio. Odds of having CS for subjects in group 2 and group 3 are 5.3 and 15.1 times the odds for subjects in group 1, respectively. Applying multivariate regression analysis to the largest series of brachial artery injuries to date, we have developed a predictive scoring model of CS. PMID- 19131715 TI - Sarcoma in the forearm and hand: clinical outcomes and microsurgical reconstruction for limb salvage. AB - Sarcomas in the forearm and hand are very rare, and surgical outcomes have been unclear. The aim of this study was to examine oncologic outcome, microvascular reconstruction, and functional evaluation. A retrospective review was performed in 19 patients who were treated for soft tissue or osseous sarcoma. All 12 patients with sarcoma in the forearm were treated with limb salvage and needed microvascular reconstruction. Flap survival was excellent. Local recurrence occurred in 4 patients, 3 of which had previously undergone inadequate resection. Three patients had distant recurrence and 1 died of disease. The mean Enneking functional score was 83% at a mean follow-up period of 37 months. Although sarcomas often metastasize, the overall survival rate is excellent. Wide marginal resection during initial surgery is the most predictive factor for tumor control. Microvascular cutaneous, myocutaneous, or osteocutaneous flap reconstruction is essential for limb salvage and provides reliable, safe coverage with reasonable preservation of function. PMID- 19131716 TI - Heterodigital vascular island flap for simultaneous resurfacing and revascularization of digits. AB - Devascularized digits with segmental skin and vessel loss require revascularization and resurfacing. This can be addressed by using a heterodigital vascular island flap. This flap brings with it an appropriately sized pristine artery of optimal length for revascularization and provides simultaneous skin cover. We describe 3 cases. Primary wound healing was achieved in all patients with good functional recovery and acceptable donor site morbidity. We compare the options available for reconstructing such defects and discuss other possible surgical indications for this flap. PMID- 19131717 TI - Intravascular stenting (IVaS) method for fingertip replantation. AB - Remarkable progress has been made in microsurgery. However, fingertip replantation following amputation has not gained much popularity because of its technical difficulty. We have developed the intravascular stenting (IVaS) method, in which a nylon monofilament is placed inside the vessel lumen to act as a temporary stent, facilitating anastomosis completion. This report describes 7 fingertip replantations using the IVaS method. Intravascular stent size varied from 4-0 to 6-0 (0.199-0.07 mm diameter). There were no cases in which the back wall of a vessel became inadvertently caught in the anastomosis. The overall survival rate for distal digital replants was 85% (6/7 replants). It is very difficult to evenly anastomose vessels of differing diameter, especially on a supermicrosurgical scale. In this respect, the IVaS method plays a role in stably anchoring the 2 vessel ends, allowing for the even spacing of suture knots, even in vessels of different caliber. Because of its ease of use and exactitude, many surgeons may be able to use the IVaS method to reliably complete small anastomoses in fingertip replantations. PMID- 19131718 TI - Perfusion dynamics of free DIEP and SIEA flaps during the first postoperative week monitored with dynamic infrared thermography. AB - Perfusion dynamics of 16 free DIEP flaps and 4 free SIEA flaps were studied during the first, third, and sixth postoperative day using dynamic infrared thermography (DIRT). For both flap types the zone positioned over the perforator is perfused first, followed by the adjacent ipsilateral zone, and finally the contralateral zones. Perfusion of the subdermal plexus of all zones preceded the perfusion of the subcutaneous layer of all zones. While the initial hyperemia subsided with time, the total number of hot spots increased with time. Perfusion of free DIEP and SIEA flaps during the first postoperative week is a dynamic process. The perfusion shows a stepwise progression at the level of the subdermal plexus and at the level of the subcutaneous layer each with its own time sequence and with the midline as an area of resistance for circulation. PMID- 19131719 TI - Biometric morphing: a novel technique for the analysis of morphologic outcomes after facial surgery. AB - The results of facial surgery are intuitively judged in terms of the visible changes in facial features or proportions. However, describing these morphologic outcomes objectively remains a challenge. Biometric morphing addresses this issue by merging statistical shape analysis and image processing. This study describes the implementation of biometric morphing in describing the average morphologic result of facial surgery. The biometric morphing protocol was applied to pre- and postoperative images of the following: (1) 40 dorsal hump reduction rhinoplasties and (2) 20 unilateral enophthalmos repairs. Pre- and postoperative average images (average morphs) were generated. The average morphs provided an objective rendering of nasal and periorbital morphology, which summarized the average features and extent of deformity in a population of patients. Subtle alterations in morphology after surgery, which would otherwise be difficult to identify or demonstrate, were clearly illustrated. Biometric morphing is an effective instrument for describing average facial morphology in a population of patients. PMID- 19131720 TI - Functional outcomes and reevaluation of esophageal speech after free jejunal transfer in two hundred thirty-six cases. AB - Swallowing and communication are occasionally impaired after free jejunal transfer. Here, the relationship between surgical procedure and functional outcome was analyzed in 236 patients undergoing free jejunal transfer after total laryngopharyngectomy from 1992 through 2003. Swallowing and communication functions were also investigated with a questionnaire in 40 long-surviving patients. Although oral feeding could be resumed after surgery in most patients, anastomotic stricture and nasal regurgitation occurred in 12.7% and 29.7% of patients, respectively. Use of our standardized procedure, the tensed jejunal method, significantly reduced the incidence of stricture (P < 0.01) but increased the rate of nasal regurgitation; however, in most cases regurgitation gradually resolved. Of the 40 long-surviving patients, 17 attended a speech rehabilitation program at which 12 learned to perform esophageal speech without voice restoration procedures (11 of the 12 had received a tensed jejunal graft). Our standardized procedure helps prevent strictures and encourages esophageal speech. PMID- 19131721 TI - Comprehensive reconstruction of the maxilla after a failed premaxillary osteotomy: a case report with long-term follow-up. AB - Major complications after maxillary orthognathic surgery are fortunately uncommon when executed properly. However, segmental necrosis has been reported. When this occurs, comprehensive reconstruction of the maxilla becomes difficult because both hard and soft tissues are lost. Use of a free flap such as the fibula for this purpose is difficult because it is bulky. We report a 40-year-old woman who presented with a defect of the anterior maxilla that occurred after necrosis of the premaxilla, which we treated with a modified prefabricated osteocutaneous free fibula flap and dental implants with long-term follow-up. The modified prefabricated free fibula flap is suitable for maxillary reconstruction. PMID- 19131722 TI - Nasal dorsum reconstruction with 11th rib cartilage and auricular cartilage grafts. AB - We present a review of international literature on the topic of nasal dorsum reconstruction with 11th rib cartilage and auricular cartilage grafts, analyzing 123 patients selected from 653 cases of rhinoplasties performed between January 1990 and October 2007 at the Department of Plastic and Reconstructive Surgery of the University of Rome "Tor Vergata." We present our experience with the correction of deformities of the nasal dorsum using rib cartilage and auricular cartilage grafts. The majority of the time, nasal dorsum deformities are complicated defects to correct surgically. They can be a consequence of naso ethmoid-orbital fractures and of surgical procedures in the nasal area where a loss of bone or septal cartilaginous support has occurred. After a review of the techniques employed in the reconstruction, we describe the advantage of the use of rib cartilage and our experience using this procedure. In the sample examined, 84% of treated patients showed cosmetic improvements, with satisfactory results to both surgeon and patient. A functional improvement has been achieved in 94% of the operated cases. PMID- 19131723 TI - Palatal fistulae resulting from cocaine abuse. AB - Cocaine fistulae require repair with well-vascularized material. In the technique used this is accomplished by closure of the nasal layer by delayed palatal flaps and the oral layer with a tongue flap. Three cases are presented. PMID- 19131724 TI - The role of the plastic surgeon in dealing with soft tissue injuries: experience from the second Israel-Lebanon war, 2006. AB - During the 2006 war between Israel and Lebanon, 282 Israeli soldiers were evacuated to Rambam Health Care Campus. Of these, 210 were admitted for observation or treatment, and 15 of these were admitted to the Department of Plastic and Reconstructive Surgery. Thirty-five other soldiers, hospitalized in other departments, required the care of Plastic Surgeons, either for conservative or surgical treatment. The injury profile observed was consistent with data from previous low-intensity warfare, which demonstrated that over 80% of injuries were produced by fragmentation weapons, such as artillery, mortarshells, rockets, and missiles. It differs, however, from our experience in previous wars and our expectations regarding burn wounds, both in incidence and severity, which were significantly lower as compared with the past. This article presents our management of extensive soft tissue injuries, and details 3 representative cases. It highlights the role of the Plastic Surgeon as part of the whole treatment in this type of injury and helps to predict the needs of the medical system in preparation for the future. PMID- 19131725 TI - The contralateral rectus abdominis musculocutaneous flap for treatment of lower extremity lymphedema. AB - Lymphedema is common after inguinal lymphadenectomy or resection of groin tumors. Animal studies have shown success using the rectus abdominis musculocutaneous (RAM) flap as a treatment for lymphedema. Four patients with acquired lower extremity lymphedema were treated with a contralateral RAM flap with an inferior cutaneous pedicle left intact to facilitate lymphatic drainage into the unaffected groin. One patient also had lymphaticovenous anastomoses performed during flap transfer. All flaps survived with no postoperative complications. With a mean follow-up of 31 months, the mean reduction in limb circumference from the preoperative excess was 81% at the thigh, 70% at the calf, and 71% at the ankle. None of the patients with recurrent cellulitis had further incidences of groin cellulitis. Two patients required future flap debulking. Lymphoscintigraphy was performed in 1 patient and demonstrated reconstitution of lymphatic flow from the affected leg through the flap. According to this preliminary study, transfer of a contralateral RAM flap to the groin of a lymphedematous leg improves lymphedema and decreases the incidence of cellulitis. PMID- 19131726 TI - Normal transvaginal delivery after abdominal wall reconstruction with tensor fascia lata muscle flap. AB - A 19-year-old woman presented with a tumor in the lower abdominal wall at the site of the scar that formed after appendectomy at age 7. The tumor mass was surgically removed, and the resulting full-thickness defect of the abdominal wall was reconstructed with autogenous tensor fascia lata muscle flap. Within 10 weeks of the reconstruction, she conceived and proceeded with an uneventful pregnancy and transvaginal delivery. PMID- 19131727 TI - Merkel cell carcinoma of the lower extremity: report of four cases and new considerations. AB - Merkel cell carcinoma (MCC) is a rare neuroendocrine cutaneous malignancy that predominantly arises in the head and neck region. We describe clinical features, diagnosis, and treatment in 4 cases of MCC, presenting an uncommon female predominant occurrence and an unusual primary site: the lower limb. In all cases diagnosis was established by histopathologic examination. Primary MCC and locally recurrence disease were treated in all patients with a wide surgical excision (3 cm margin) including fascia. Lymphadenectomy was reserved for a patient with clinical evidence of nodal involvement. Both chemotherapy administered in 2 cases and radiotherapy in 1 case produced limited responses. Early diagnosis is critical because this tumor is aggressive and has a high rate of local recurrence and metastatic spread. However, its nondistinctive appearance frequently delays diagnosis and its rarity avoids an optimal treatment guideline setting. PMID- 19131728 TI - Application of AAV2-mediated bFGF gene therapy on survival of ischemic flaps: effects of timing of gene transfer. AB - Necrosis of surgically transferred flaps is a major problem in reconstructive surgery. We investigated efficacy of a new vector system-adeno-associated viral 2 (AAV2)-mediated bFGF gene transfer to enhance survival of the ischemic flap. Thirty-eight Sprague-Dawley rats were divided into 3 gene therapy groups and 1 nontreated control of 9 or 10 each. 7.5 x 10(10) AAV2-bFGF viral particles were injected to the dorsum of each of the 29 rats; these rats were divided into 3 groups according to the timing of flap elevation. At the time of surgery, 1 week, and 2 weeks after surgery, flaps of 3 x 7 cm were raised. One week after surgery, flap viability was measured. Vascularization and immunohistochemical staining of the bFGF were evaluated of histologic sections. Flap viability was significantly improved by the AAV2-bFGF gene therapy at the time of surgery, and the flaps with the greatest survival area were found in the rats injected with AAV2-bFGF, 2 weeks before surgery. However, flap viability was significantly decreased by the gene therapy 1 week before surgery. Histologically, vascularity was increased in the groups with AAV2-bFGF injection and immunohistochemical staining showed greatly enhanced bFGF expression by gene transfer. The novel approach of AAV2 bFGF gene therapy shows encouraging manifestations in improving survival of flaps when the flaps are prefabricated during or 2 weeks before surgery. PMID- 19131729 TI - Topical negative pressure stimulates endothelial migration and proliferation: a suggested mechanism for improved integration of Integra. AB - Topical negative pressure is an effective technique to promote wound healing and the integration of skin graft and synthetic dermal equivalents. We describe an in vitro model to investigate the effect of negative pressure on angiogenesis, a pivotal step. Dermal fibroblasts or human microvascular endothelial cells were cultured on Integra and subjected to intermittent or continuous negative pressure. At fixed intervals of over 120 hours, the Integra was fixed and assessed for cell migration (microscopy), cell viability (MTS assay), and cell proliferation (Ki67 immunostaining). Under control conditions, endothelial cells formed a monolayer and failed to ingress, whereas fibroblasts migrated throughout the Integra within 24 hours. Negative pressure switches endothelial cell to a migratory and proliferative phenotype. Ingress is greatest with intermittent rather than continuous negative pressure. It has no effect on dermal fibroblast function. This study identifies an important, potential pro-angiogenic mechanism by which topical negative pressure promotes wound healing. PMID- 19131730 TI - Tissue engineering in plastic surgery: an up-to-date review of the current literature. AB - Tissue engineering is an interdisciplinary field that applies the principles of engineering and life sciences toward the development of biological substitutes that restore, maintain, or improve tissue function. This field has enjoyed tremendous growth in the past 10 years fuelled by its potential role in regenerating new tissues and naturally healing injured or diseased organs. Stem cells due to their pluripotentiality and unlimited capacity for self-renewal, may allow significant advances for distinct reconstructive and cosmetic procedures. This review aims at outlining the principles of tissue engineering, focusing on the use of adult-derived stem cells as applied to the research and practice of plastic surgery. Review categories have been divided into tissue engineering of the skin and connective tissue, bone marrow, cartilage, adipose tissue, and breast tissue. An analytical review of the current literature on stem cell technology on the above mentioned areas is presented. There have been reports of side effects and unsuccessful treatments. The key to the progress of tissue engineering is an understanding between basic scientists, biochemical engineers, clinicians, and industry. Although there has been an ongoing research pointing to the enormous potential of using stem cells in cosmetic and reconstructive surgery, at this stage, stem cell therapy is still a hope that has not been fully studied and approved. More long-term studies are needed and many questions remain to be answered. PMID- 19131731 TI - Histologic analysis of keloids and hypertrophic scars. PMID- 19131732 TI - Measurement of airborne particle concentrations near the Sunset Crater volcano, Arizona. AB - Direct measurements of airborne particle mass concentrations or mass loads are often used to estimate health effects from the inhalation of resuspended contaminated soil. Airborne particle mass concentrations were measured using a personal sampler under a variety of surface-disturbing activities within different depositional environments at both volcanic and nonvolcanic sites near the Sunset Crater volcano in northern Arizona. Focused field investigations were performed at this analog site to improve the understanding of natural and human induced processes at Yucca Mountain, Nevada. The level of surface-disturbing activity was found to be the most influential factor affecting the measured airborne particle concentrations, which increased over three orders of magnitude relative to ambient conditions. As the surface-disturbing activity level increased, the particle size distribution and the majority of airborne particle mass shifted from particles with aerodynamic diameters less than 10 mum (0.00039 in) to particles with aerodynamic diameters greater than 10 mum (0.00039 in). Under ambient conditions, above average wind speeds tended to increase airborne particle concentrations. In contrast, stronger winds tended to decrease airborne particle concentrations in the breathing zone during light and heavy surface disturbing conditions. A slight increase in the average airborne particle concentration during ambient conditions was found above older nonvolcanic deposits, which tended to be finer grained than the Sunset Crater tephra deposits. An increased airborne particle concentration was realized when walking on an extremely fine-grained deposit, but the sensitivity of airborne particle concentrations to the resuspendible fraction of near-surface grain mass was not conclusive in the field setting when human activities disturbed the bulk of near surface material. Although the limited sample size precluded detailed statistical analysis, the differences in airborne particle concentration over 900-y weathered volcanic and nonvolcanic deposits appeared to be potentially significant only under heavy surface disturbances. PMID- 19131733 TI - Cancer incidence in municipalities near two former nuclear materials processing facilities in Pennsylvania--an update. AB - Previous studies of cancer incidence among persons living in municipalities within one mile of two nuclear materials processing and fabrication plants in Pennsylvania were extended for the years 1998-2004. It had been shown that mailing addresses for residents of rural areas often did not reflect the actual municipality of residence and, if not corrected, would bias study results. The previous studies had corrected for this bias. Accordingly for the extended study, we obtained mailing addresses from the Pennsylvania Department of Health (PDH) for 866 persons with cancer who presumably lived in one of eight minor civil divisions (MCDs) near or encompassing the former nuclear facilities, designated as Area 1 in previous studies conducted by the PDH. Street addresses were geocoded and local postmasters were asked to place rural delivery addresses, post office boxes and street addresses that could not be geocoded into the correct MCD of actual residence. Over 15% of the mailing addresses were found not to be within the boundaries of the Area 1 municipalities. After the mailing addresses of individuals with cancer were placed in their proper MCD of residence, the number of persons diagnosed with cancer (n = 708) and confirmed to have lived in Area 1 was as expected (728.4) based on cancer incidence rates in the general population of Pennsylvania (SIR 0.97; 95% CI 0.90-1.05). To further evaluate the patterns of cancer rates near these nuclear facilities and the influence of improved reporting and geocoding of addresses over time, analyses were conducted of publicly available cancer incidence data from 1990 through 2004. Based on mailing addresses, a steady decrease in the number of cancers reported in the Area 1 proximal MCDs was seen, in contrast to a steady rise in the number of cancers reported in seven adjacent but more distant MCDs from the nuclear facilities, designated as Area 2. These patterns were attributed to improvements over time in the geocoding of residential mailing addresses coupled with the gradual elimination and replacement of rural delivery addresses with street addresses. The incorrect placement of mailing addresses in residential Area 1 municipalities prior to about 2002 overestimated the number of cancers occurring among residents living in close proximity to the nuclear facilities and, correspondingly, underestimated the number among Area 2 residents. Summing Area 1 and Area 2 data showed that there was no change in cancer rates over time. These results are consistent with previous studies indicating that living in municipalities near the former Apollo-Parks nuclear facilities was not associated with an increase in cancer occurrence. PMID- 19131735 TI - An evaluation of fluoroscopic times and peak skin doses during radiofrequency catheter ablation and biventricular internal cardioverter defibrillator implant procedures. AB - Fluoroscopy-guided medical procedures are generally diagnostic in nature and involve low dose and minimal risk to the patient. However, an increasingly larger percentage of procedures offer a therapeutic benefit to the patient. Such interventional procedures are very often technically difficult, but offer a less invasive alternative to higher-risk surgical procedures. Although infrequent, severe skin injury can result from these procedures that involve total fluoroscopy time in excess of 1 h. The purpose of this study was to evaluate fluoroscopy time and peak skin dose (PSD) for lengthy fluoroscopy procedures, and to determine the variables that may best predict PSD. PMID- 19131734 TI - County mortality and cancer incidence in relation to living near two former nuclear materials processing facilities in Pennsylvania--an update. AB - A previous county mortality study of populations living near two nuclear materials processing and fabrication facilities in Westmoreland and Armstrong counties in Pennsylvania (1950-1995) was extended through 2004. Noncancer mortality (1996-2004) and cancer incidence (1990-2004) were also evaluated. Among the Westmoreland and Armstrong populations, 10,547 cancer deaths occurred during the period 1996 through 2004 and the relative risk (RR) based on comparisons with six demographically similar counties in western Pennsylvania was 0.97, that is, almost exactly as expected, and no different from our previously published analyses covering the years 1950-1995. The results based on cancer incidence data were very similar to those based on cancer mortality data. Over the years 1990 though 2004, 39,350 incident cancers were reported among residents of Armstrong and Westmoreland counties and the RR based on the six demographically similar counties was 0.99, that is, almost exactly as expected. The number of deaths from nonmalignant conditions was 36,565 and very close to the number expected (RR 1.01). Overall, no increases in cancer or nonmalignant diseases could be attributed to living in counties with nuclear materials processing and fabrication facilities. PMID- 19131736 TI - Modeling the imprecision in prospective dosimetry of internal exposure to uranium. AB - The dosimetry of internal exposure to radionuclides is performed on the basis of biokinetic and dosimetric models. For prospective purpose, the organ or effective dose resulting from potential conditions of exposure can be calculated by applying these models with dedicated software. However, it is acknowledged that a significant uncertainty is associated with such calculation due to the variability of individual cases and to the possible lack of knowledge about some factors influencing the dosimetry. This uncertainty has been studied in a range of situations by modeling the uncertainty on the model parameters by probability distributions and propagating this uncertainty onto the dose result by Monte Carlo calculation. However, while probability distributions are well adapted to model the known variability of a parameter, they may lead to an unrealistically low estimate of the uncertainty due to a lack of knowledge about some input parameters. Here we present a mathematical method, based on the Dempster-Shafer theory, to deal with such imprecise knowledge. We apply this method to the prospective dosimetry of inhaled uranium dust in the nuclear fuel cycle when its physico-chemical properties are not precisely known. The results show an increased estimation of the range of uncertainty as compared to the application of a probabilistic method. This Dempster-Shafer method may valuably be applied in future prospective dosimetry of internal exposure in order to more realistically estimate the uncertainty resulting from an imprecise knowledge of the parameters of the dose calculation. PMID- 19131737 TI - Analytical and Monte Carlo assessment of activity and local dose after a wound contamination by activation products. AB - The activity and local dose following a right index finger wound contamination by activation products are assessed. Measurements with a high purity germanium detector and a four positions measurement protocol enabled a better localization of the contaminant source. From the source location and detector calibration, the remaining wound activity and local absorbed dose were deduced. An analytical model, based on a two dimensional simplification of the problem, is presented. It is shown to provide a fast and quite accurate activity assessment when the contaminants are described as a point source. The contaminants' location and activity were then more accurately assessed using Monte Carlo calculations based on the OEDIPE software and a voxelized phantom of the index finger. Describing the contaminant mixture as a point source resulted in an agreement of experimental and computed data around 6% for most of the radionuclides. The total activity, due to 11 radionuclides, was estimated to be (9.5 +/- 0.4) kBq at measurement day. Since the point source is found to be less than 1 mm under the skin, the equivalent skin dose is calculated and found to be around 680 mSv in the first year after the contamination, and this value decreases to 250 mSv in the second year. The relevance of equivalent skin dose as an estimate of the sanitary risk is discussed, and it is concluded that for this case it gives the upper end estimate of the risk. PMID- 19131738 TI - Scaled tests and modeling of effluent stack sampling location mixing. AB - A three-dimensional computational fluid dynamics computer model was used to evaluate the mixing at a sampling system for radioactive air emissions. Researchers sought to determine whether the location would meet the criteria for uniform air velocity and contaminant concentration as prescribed in the American National Standards Institute standard, Sampling and Monitoring Releases of Airborne Radioactive Substances from the Stacks and Ducts of Nuclear Facilities. This standard requires that the sampling location be well-mixed and stipulates specific tests to verify the extent of mixing. The exhaust system for the Radiochemical Processing Laboratory was modeled with a computational fluid dynamics code to better understand the flow and contaminant mixing and to predict mixing test results. The modeled results were compared to actual measurements made at a scale-model stack and to the limited data set for the full-scale facility stack. Results indicated that the computational fluid dynamics code provides reasonable predictions for velocity, cyclonic flow, gas, and aerosol uniformity, although the code predicts greater improvement in mixing as the injection point is moved farther away from the sampling location than is actually observed by measurements. In expanding from small to full scale, the modeled predictions for full-scale measurements show similar uniformity values as in the scale model. This work indicated that a computational fluid dynamics code can be a cost-effective aid in designing or retrofitting a facility's stack sampling location that will be required to meet standard ANSI/HPS N13.1-1999. PMID- 19131739 TI - Lung fibrosis and lung cancer incidence in beagle dogs that inhaled 238PuO2 or 239PuO2. AB - Determination of radiation protection guidelines for persons working with plutonium has been complicated by limited human data on the biological behavior and subsequent health effects from internally deposited plutonium. One solution has been the use of animal models to predict likely health effects in humans. To compare the relationships between plutonium inhalation and lung fibrosis and lung cancer, data from life-span studies of beagle dogs given a single exposure to either plutonium-238 dioxide (238PuO2) or plutonium-239 dioxide(239PuO2) were analyzed. Estimates of the cumulative hazard of lung fibrosis and lung cancer after exposure to either were generated. The hazard of lung fibrosis was not consistent with a linear no-threshold model, although the magnitude of the threshold differed by radionuclide. In dogs given 239PuO2,the best model of lung fibrosis incorporated a linear dose response function; a linear-quadratic dose response function fit the data better in dogs given 238PuO2. At any given cumulative dose, the lung fibrosis hazard was greater for dogs given 238PuO2. In dogs given 238PuO2, with or without covariates, aquadratic dose-response function for lung cancer hazard fit better than a linear no-threshold model. In dogs given 239PuO2, models of lung cancer with the dose-response function as the sole predictor variable were consistent with a linear no-threshold model; however, a quadratic dose-response function with a cell-killing term fit better. These findings have implications for radiation protection because, while lung cancer hazard was dependent on cumulative dose, regardless of isotope, the lung fibrosis hazard depended on both cumulative dose and isotope. PMID- 19131741 TI - Overestimation of thyroid cancer incidence after Chernobyl. PMID- 19131742 TI - Alliance of Nuclear Worker Advocacy Groups. PMID- 19131747 TI - International Anesthesiology Clinics. Preface. PMID- 19131748 TI - Cardiopulmonary to cardiocerebral resuscitation: current challenges and future directions. PMID- 19131749 TI - Renal replacement therapy in the intensive care unit. PMID- 19131750 TI - Trauma. PMID- 19131751 TI - Acute spinal cord injury. PMID- 19131752 TI - Management of sepsis in the ICU. PMID- 19131753 TI - Corticosteroids in the ICU. PMID- 19131754 TI - Adult respiratory distress syndrome. PMID- 19131755 TI - Pulmonary hypertension and right ventricular function: interdependence in pathophysiology and management. PMID- 19131756 TI - Evidence-based nutritional support in the intensive care unit. PMID- 19131757 TI - Glycemic control in critical care: current benefits and future needs. PMID- 19131758 TI - Tele-ICU: a new paradigm in critical care. PMID- 19131760 TI - Multifunctional in situ photopolymerized semi-interpenetrating network system is an effective donor site dressing: a cross comparison study in a swine model. AB - Effective dressings for donor sites or other partial thickness wounds must promote removal of nonviable or necrotic tissue, eradication and prevention of microbial infiltrate, exudate absorbance, and regrowth of healthy epidermis and dermis. There are many commonly used products that facilitate these processes. Established properties of an in situ photopolymerizable semi-interpenetrating network (sIPN) suggest that it is also a viable treatment option. The widely varying material properties suggest that these dressing treatments may elicit different healing responses via different cellular mechanisms. In this study, we sought to resolve the differences in healing between Acticoat, sIPN, nonadherent dressing with Tisseel, and Xeroform dressing treatments in a porcine partial thickness wound model. Donor site wounds were produced on pigs at two cut depths and dressed with Acticoat, sIPN, nonadherent dressing with Tisseel, and Xeroform with alternatively placed autografts to provide a control area between each test site. Pigs were euthanized at 4, 7, 14, and 42 days for macroscopic examination and biopsy collection. Biopsies were analyzed histologically by two blinded observers for cellular densities and regional thicknesses within the tissue. sIPN and Xeroform-treated wounds were healed by 7 days, and Acticoat- and nonadherent dressing with Tisseel-treated wounds were healed by 14 days. Inflammatory responses were between comparable treatment type across all time periods. Dermal granulation features increased with time but were not significantly different. All dressing treatments elicited wound healing without outstanding toxicity or pathology indicating that sIPN is a comparable and viable treatment for partial thickness wounds. PMID- 19131761 TI - Response to a nursing-driven protocol for sedation and analgesia in a burn-trauma ICU. AB - A nursing-driven protocol for sedation and analgesia for our burn unit was developed based on Society of Critical Care Medicine recommendation. Our goal was to evaluate nursing staff perceptions of how the protocol affected their ability to achieve care goals for their patients. From March 1 through August 31, 2007, nursing staff were surveyed at the conclusion of each shift when they cared for a patient on protocol. Three hundred and thirty of 506 possible surveys were completed. The preponderance of respondents was nurses with less than 3 years of clinical experience. Sedation and analgesia holidays were reported as performed per protocol 68% of the time, and 80% of deviations from the protocol holiday were because the patient was already at or above target Riker and Visual Analog Scale scores. The majority of responding nurses agreed or strongly agreed that the protocol was effective (62%), that it allowed them to use their clinical assessment skills (64%), and that the protocol was clearly written (70%). Level of comfort with performing a holiday was inversely associated with years of nursing experience (chi, P = .02). Nursing interest in routinely using the protocol was also inversely associated with years of nursing experience (chi, P = .001). Nursing staff responded positively to the introduction of the nursing driven protocol. Nurses with less clinical experience were more comfortable with analgesia and sedation holidays and were more supportive of consistently using the protocol for mechanically ventilated patients. Further education may help foster support from more senior nursing staff. PMID- 19131762 TI - Health care costs in gout: what are these emerging data telling us? PMID- 19131763 TI - Repeat injections of intra-articular botulinum toxin a for the treatment of chronic arthritis joint pain. PMID- 19131764 TI - Calcium crystals and arthritis: what is new under polarizing light? PMID- 19131765 TI - Primary bone lymphoma presenting as a monoarthritis. PMID- 19131766 TI - Giant cell arteritis: a rare disease in Asians. PMID- 19131768 TI - Tired, achy, and overweight, the inflammatory nature of obesity. PMID- 19131769 TI - Neonatal primary hemophagocytic lymphohistiocytosis in Turkish children. AB - Although the data on hemophagocytic lymphohistiocytosis (HLH) has gradually increased, the neonatal-onset HLH patients have usually been reported as case reports or together with other age groups of patients. The aim of this study was to draw attention to the clinical and laboratory characteristics of neonatal HLH cases. Herein, the data of 8 primary, neonatal-onset HLH patients are reported. Mutational analyses were performed in 7 of the patients and mutations in UNC13D gene were detected in 3 of the patients, whereas 2 patients were found to have perforin gene mutation. Four of the patients were symptomatic within the initial 10 days of life. One patient with perforin mutation (1122 G>A) had a very severe clinical course and died on the seventh day of life before receiving any specific treatment. Another patient with UNC13D 2783 G>C, who became symptomatic on the sixth day of life, underwent early hematopoietic stem cell transplantation and is currently alive at 8 years of age. Two of these 4 patients had extensively high serum ferritin levels mimicking neonatal hemochromatosis. Of the 4 patients who became symptomatic after 20th day of newborn period, 1 was found to have perforin gene mutation (445 G>A) and 2 siblings were detected to have a missense mutation in UNC13D (640 C>T) gene. The latter patients with UNC13D mutations could survive 3 and 4 months, although their parents ceased therapy. The patient with perforin mutation survived 11 months. PMID- 19131770 TI - Chronic granulomatous disease (CGD) mimicking neoplasms: a suspected mediastinal teratoma unmasking as thymic granulomas due to X-linked CGD, and 2 related cases. AB - Patients with chronic granulomatous disease (CGD) suffer from severe bacterial and fungal infections and deregulated inflammation, which are often associated with granuloma formation. We describe a 2-year-old boy who was seemingly healthy at the age of 1 year when a conventional chest radiograph was taken to exclude pulmonary aspiration of a piece of apple. Incidentally, a space-occupying mediastinal mass was revealed that was further evaluated by magnetic resonance imaging. Varying solid and also cystic, septated parts of the mass could be discerned and it was considered to be a teratoma. Removal of the mass by surgery was arduous because of adhesiveness to surrounding areas and led to severe complications. Unexpectedly, histopathologic examination revealed massive acute granulomatous inflammation with liquefied thymic cysts. X-linked CGD was subsequently diagnosed by a dihydrorhodamine 123 assay and sequencing of the CYBB gene (hotspot mutation c.742-743insA). This is the third example that we are aware of, where CGD granulomas were mistaken for neoplasms. The other 2 patients were initially believed to have tumors of the stomach and the urinary bladder, respectively. All patients initially received inadequate treatment. We discuss possible strategies to avoid such tragic confusions. PMID- 19131771 TI - PTX3 as a potential novel tool for the diagnosis and monitoring of pulmonary fungal infections in immuno-compromised pediatric patients. AB - Pentraxin-3 (PTX3) is a member of the long pentraxin superfamily and has a nonredundant role in mediating resistance to fungal pathogens. Serial monitoring of PTX3 plasmatic levels was performed in 10 pediatric leukemia patients affected by pulmonary fungal infections. When compared with values of a control pediatric cohort, PTX3 showed significantly higher plasmatic values. Moreover, the response to the antifungal therapy correlated with normalization of PTX3 values. PTX3 may represent a useful tool for the diagnosis and monitoring of fungal infections in immuno-compromised children. PMID- 19131772 TI - The clinical feature of invasive fungal infection in pediatric patients with hematologic and malignant diseases: a 10-year analysis at a single institution at Japan. AB - Invasive fungal infections (IFI) are an important complication in hematologic malignancies and stem-cell transplantation (SCT). However, there are limited data characterizing IFI in children. The clinical feature of IFI after chemotherapy and SCT were analyzed in 334 pediatric patients treated at Hokkaido University Hospital from 1997 to 2006. The cumulative incidence of IFI was 6.9%; this comprised cases of proven, probable and possible IFI at rates of 1.2%, 3.0%, and 2.7%, respectively. The infected lesions were lung in 14 patients, liver in 5 patients, brain in 3 patients, fungemia in 2 patients, kidney in 1 patient, and endophthalmitis in 1 patient. The mortality of IFI was 48.2%, excluding patients who died due to relapse and interstitial pneumonitis; in particular, 71.4% patients with a lung lesion (10/14) died due to IFI. Fifty-nine pediatric patients died in our institution over the 10-year period of the study and IFI was the direct cause of death in 18.6% (11/59) of the patients. Risk factors for IFI with chemotherapy and SCT were also analyzed. Univariate analysis showed that age at diagnosis older than 10 years, relapse of original disease, long-term administration of broad-spectrum antibiotics, and acute myelogenous leukemia (AML) were the risk factors for IFI. All patients with IFI received long-term antibiotic therapy. AML was most strongly associated using a multivariate analysis. The prognosis of IFI has been expected poor; therefore, prevention of this condition, especially for older patients with AML, would be important. PMID- 19131773 TI - Metastatic malignant peripheral nerve sheath tumor in a newborn. AB - Malignant peripheral nerve sheath tumors are rare tumors, especially in the newborn period. Diagnosis is based on clinical findings, radiography, and fine needle biopsy or tissue sampling. Ideal management is controversial and extremely difficult. The survival rate is extremely low. We present a case of a newborn in which this tumor is diagnosed in the first few days of life as an ulcerating tumor imbedded in the entire forefoot with already metastatic spread to the lymph nodes and abdomen. Questions arose on the best treatment in postpartum life. Below the knee amputation was performed for nursing purposes, but eventually a no intention-to treat policy was chosen by the multidisciplinary team and parents. Detection of soft tissue tumors in the prenatal period is difficult and not as easy as for other structural defects. Would prenatal detection have changed the course of life of this young infant? Will future opportunities for detection of soft tissue tumors occur? Diagnostic and therapeutic considerations are discussed alongside the present literature. PMID- 19131774 TI - Significance of fetal hemoglobin values in detection of heterozygotes in fanconi anemia: reevaluation of fetal hemoglobin values by a sensitive method. AB - Fanconi anemia (FA) is a genetically very heterogeneous disease making routine detection of carriers quite difficult by molecular analysis. Finding alternative method has vital importance especially in populations where prevalence of the disease is quite high because of consanguineous marriages. The aim of this study was to find a considerably reliable parameter to detect FA carriers by methods other than molecular analysis. The subjects of this study were 66 parents of children with FA and 40 age and sex compatible individuals from the normal healthy population. An index family with a known mutation was also included as an evidence for verification of the results. The mean fetal hemoglobin (HbF) values (0.81%+/-0.72%) of FA heterozygotes studied by high-performance liquid chromatography was significantly higher than that (0.37%+/-0.32%) of the control group (P<0.001). Additionally, there was a positive correlation between the HbF value of the children (mean: 4.50+/-1.59) and the parents (mean: 0.81+/-0.72) (r: 0.698, P=0.01). No significant difference was detected between the hemoglobin, hematocrit, red blood cell count, mean corpuscular volume, red cell distribution width, white blood cell count, absolute neutrophil count, and platelet counts of the study and control groups. The results of this study suggest that the HbF values may be used as a marker to predict carriers in the family members of a child with FA when definitive diagnosis by molecular analysis is not possible. PMID- 19131775 TI - Pediatric hibernoma: a case review. AB - We present an unusual case of supraclavicular benign hibernoma in a 12-year-old girl who presented with chest discomfort, night sweats, shortness of breath, fatigue, and pruritus. This is the first case of hibernoma being reported as symptomatic in the world literature. Symptoms persisted for 1 year till excision biopsy. Biopsy of supraclavicular lump revealed a hibernoma. Symptoms resolved within 2 months of excision and 1-year follow-up showed no recurrence of symptoms or supraclavicular swelling. PMID- 19131776 TI - Prevalence of multiple health-related behaviors in adolescents with cancer. AB - OBJECTIVE: This preliminary study investigated prevalence rates of multiple health-related behaviors (ie, tobacco, alcohol, and other drug use; sexual risk taking; nutrition/physical activity; overweight and dietary behaviors; sun safety) among 42 adolescents on active treatment for cancer, as compared with healthy adolescent norms. METHODS: Health-related behaviors were assessed using the 2005 National Youth Risk Behavior Survey, for which healthy adolescent norms were publicly available. RESULTS: Adolescents with cancer reported significantly lower current and lifetime rates of tobacco, alcohol, and other drug use; fruit/vegetable consumption; physical activity; and dietary behavior, in addition to lower rates of lifetime sexual intercourse, early-onset sexual intercourse, and alcohol/drug use before last sexual intercourse, compared with healthy peers. Among those who have previously engaged in sexual intercourse, there appeared to be a trend toward increased partners, current sexual activity, and lack of protection at last episode of sexual intercourse. Adolescents with cancer also reported significantly higher rates of television watching compared with healthy peers. CONCLUSIONS: Adolescents on active treatment for cancer are engaging in multiple health-risk behaviors, including sedentary behavior, poor nutrition, lack of sun safety, and sexual risk-taking (eg, multiple partners, lack of protection at last sexual intercourse). Health promotion interventions are needed during active treatment to facilitate the acquisition of good health practices as adolescents transition into survivorship. PMID- 19131777 TI - Neoadjuvant chemotherapy for osteosarcoma of the extremities in preadolescent patients: the Rizzoli Institute experience. AB - Medical records of 133 patients, 10 years old or younger with primary high-grade nonmetastatic osteosarcoma of the extremities treated at the Rizzoli Institute between 1983 and 1999 with neoadjuvant chemotherapy were reviewed and compared with those of 782 patients aged 11 to 40 years treated in the same period with the same chemotherapy protocols. In comparison to the older group, the younger group had more females, more patients with normal lactic dehydrogenase levels, and more non-limb-salvage procedures (amputation or rotationplasty). Five-year event-free and overall survivals were essentially the same in the two groups (63% and 71% vs. 60% and 70%) as were the patients rescued after relapse and presently event-free (18% vs. 20%). The authors conclude that there does not seem to be any indication to treat preadolescent primary high-grade nonmetastatic osteosarcoma patients by alternative and/or more aggressive therapies. PMID- 19131778 TI - Celiac disease in a child with beta-thalassemia major: a need for improved screening and awareness. AB - Growth failure is one of the most common problems in children with thalassemia with multiple etiologies. We present a case of celiac disease, an underdiagnosed cause of growth failure in a child with beta-thalassemia major. A 10-year-old boy on a hypertransfusion regimen was referred for early onset growth failure. Serology for hepatitis B, hepatitis C, and HIV was negative. Serum zinc levels were normal. Thyroid function tests and growth hormone secretion, evaluated with clonidine stimulation test were normal. Malabsorption syndrome was suspected, even in the absence of gastrointestinal symptoms. Tissue transglutaminase were highly raised >300 IU/mL (normal values <15 U/L). Characteristic mucosal lesions on jejunal biopsy confirmed the diagnosis of celiac disease. Institution of a gluten-free diet resulted in rapid gain in weight and improvement in height velocity. PMID- 19131779 TI - An uncommon presenting sign of Langerhans cell histiocytosis: focal perianal lesions without systemic involvement. AB - A 3-year-old boy presented with constipation and perianal lesions resembling condyloma latum. The results of a biopsy of the perianal lesions confirmed the diagnosis of Langerhans cell histiocytosis (LCH). Although uncommon, LCH may involve the perianal region. In patients with functional constipation associated with perianal lesions that do not respond to conventional treatment, LCH should be part of the differential diagnosis. A simple biopsy of the external lesion and histologic examination of the mass are essential for diagnosis. PMID- 19131780 TI - Early detection of breakthrough trichosporonosis by serum PCR in a cord blood transplant recipient being prophylactically treated with voriconazole. AB - An 18-year-old man with myelodysplastic syndrome after stem-cell transplantation developed breakthrough trichosporonosis while being administered voriconazole, a broad-spectrum triazole. Importantly, Trichosporon was detected soon after infection by a screening serum polymerase chain reaction test, but not by blood culture. This case illustrates that the serum polymerase chain reaction assay might be effective for early detection of fungemia in immunocompromised patients, including recipients of stem-cell transplants undergoing prophylactic antifungal therapy using voriconazole, which cannot be assumed to always provide complete protection against Trichosporon infection. PMID- 19131782 TI - Acute Wilson disease associated with E beta-thalassemia. AB - We report on an 11-year-old girl with concomitant Ebeta thalassemia (EbetaT) and Wilson disease (WD). She was diagnosed with EbetaT at 2 years of age, but the coexistence of WD could only be established at 11 years. The diagnosis of the later was based on the clinical presentation of hepatitis and severe Coomb's negative hemolytic anemia, coupled with laboratory evidence of WD. To our knowledge, this is the first report on the cooccurrence of EbetaT and WD. As both the conditions are associated with variable degrees of hemolysis, WD in the setting of EbetaT may remain masked and consequently remain undiagnosed for a long time. Sudden, severe hemolysis in a patient of thalassemia may be explained by the coexistence of additional pathology, in this case WD. PMID- 19131781 TI - IL-1RA agonist (anakinra) in the treatment of multifocal castleman disease: case report. AB - Multicentric Castleman disease (MCD), or multicentric angiofollicular lymph node hyperplasia, is an idiopathic lymphoproliferative disorder that is only seldom reported in children. The clinical and laboratory findings that characterize this peculiar disorder are thought to result from increased interleukin (IL)-6 production. We report herein the case of a 13-year-old boy with a long history of hectic fevers and abdominal pain, accompanied by stunted growth and elevated biologic markers of inflammation. Surgical biopsies of a pancreatico-splenic mass and of mesenteric lymph nodes revealed mixed-type MCD, which was diagnosed 6 years after the first clinical symptoms appeared. He received combination chemotherapy (cyclophosphamide, vinblastine) associated with a monoclonal B-cell antibody (Rituximab). This treatment was well tolerated but ineffective. Given the reported success of IL-1 blocking agents for treating Still disease, another IL-6 linked disorder, we attempted to treat him with anakinra, an IL-1RA agonist. His overall state normalized and both his clinical and biologic signs dramatically improved. This is the first report of anakinra treatment for MCD. We conclude that anti-IL-1 blocking agents could be an interesting treatment alternative for MCD, a chronic debilitating disease, which still carries a poor prognosis. PMID- 19131783 TI - Severe neurologic complication after delayed hemolytic transfusion reaction in 2 children with sickle cell anemia: significant diagnosis and therapeutic challenges. AB - Although delayed hemolytic transfusion reaction (DHTR) has been widely recognized as a serious complication of red blood cell transfusion in patients with sickle cell disease (SCD), there is no consensus on its optimal management. Discontinuation of transfusion is recommended, whereas corticosteroids and immunoglobulins are considered to be beneficial. We report 2 children with sickle cell anemia who were diagnosed with DHTR and experienced a subsequent neurologic event in the course of treatment with corticosteroids. The role of corticosteroids as possible precipitating factors of neurologic complications is discussed. Pending a better understanding of the chain of events of DHTR, SCD children with DHTR should receive steroids with great caution. PMID- 19131784 TI - Visceral varicella zoster virus (VZV) after allogeneic hematopoietic stem cell transplant (HSCT) in pediatric patients with chronic graft-versus-host disease (cGVHD). AB - Reactivation of latent varicella zoster virus is one infectious complication associated with the extensive immunosuppression necessary for hematopoietic stem cell transplant. Most cases are limited to skin and mortality is low. Isolated visceral zoster is rare, presenting with ileus/abdominal pain, hepatitis, and/or hyponatremia. We present 2 cases of visceral varicella zoster virus in adolescents with chronic graft-versus-host disease after hematopoietic stem cell transplant. Both presented with elevated liver enzymes, severe abdominal pain, and hyponatremia but lacked cutaneous involvement. Both received high-dose acyclovir and showed improvement, but eventually expired from hepatic failure. The diagnosis of visceral zoster can be difficult especially without cutaneous manifestations. Vigilance is necessary in patients with chronic graft-versus-host disease, abdominal pain, and/or hepatitis and antiviral therapy should be initiated promptly. PMID- 19131785 TI - Intracranial reparative giant cell granuloma secondary to cholesteatoma in a 15 year-old girl. AB - A 15-year-old girl presented with pain in the right ear and acute onset of total peripheral facial nerve palsy. Imaging studies revealed an intracranial mass of the right temporal bone causing temporal lobe displacement. A first biopsy led to the diagnosis of intracranial giant cell reparative granuloma (GCRG), a rare benign tumor of the bone or soft tissue that can show expansive growth. The etiology of GCRG is unclear. Facial nerve palsy responded to treatment with diclofenac and physiotherapy, however, the intracranial lesion progressed at follow-up. Total surgical excision revealed a cholesteatoma central to the GCRG originating from the right middle ear and mastoid. Cholesteatoma should be considered as a trigger for intracranial GCRG growth, especially if adjacent to the temporal bone. PMID- 19131786 TI - Nasal-type NK/T-cell lymphoma presenting as hemophagocytic syndrome in an 11-year old Mexican boy. AB - Recently established by the World Health Organization classification, "nasal" and "nasal-type" NK/T-cell lymphoma arise from natural killer (NK) cells. They have distinct clinicopathologic features, specific genotype and phenotype, and a high association with Epstein-Barr virus infection. Nasal-type NK/T-cell lymphoma arise from extranasal sites including skin, soft tissue, gastrointestinal tract, liver, spleen, testes, lung, and central nervous system. Most cases are reported from Asia and South America in adults. There are very few cases reported in the pediatric age group. We report an 11-year-old child with extranodal nasal-type NK/T-cell lymphoma presenting as hemophagocytic syndrome and multiorgan system failure with fatal outcome. PMID- 19131787 TI - Differential expression of Ikaros isoforms in monozygotic twins with MLL rearranged precursor-B acute lymphoblastic leukemia. AB - Infant leukemia associated with rearrangement of the MLL gene typically presents with high-risk clinical features. Relapse is common despite aggressive therapy and perturbations in signaling pathways may contribute to disease resistance. We evaluated twin 4-month-old monozygotic baby boys who presented with MLL rearranged precursor-B acute lymphoblastic leukemia. Two different MLL/AF4 variants were found in both the twins, the first involving MLL intron 8 and AF4 intron 3 and the second stemming from translocations of MLL exon 10 and AF4 exon 4. We detected expression of the DNA-binding Ikaros isoforms, Ik1, Ikx+, Ik2 and the dominant-negative Ik4 Ikaros isoform in both patients. However, the dominant negative Ik8 isoform was detected in only 1 boy, suggesting a common genetic ontogeny that was modulated by leukemic evolution. Further exploration of Ikaros expression in the background of MLL rearrangements may provide new insights into disease pathogenesis and could offer targets for novel chemotherapeutic agents. PMID- 19131788 TI - Tacrolimus with mini-methotrexate as prophylaxis for graft-versus-host disease in pediatric patients after allogeneic peripheral blood stem cell transplant or bone marrow transplant. AB - Experience with tacrolimus in combination with mini-methotrexate to prevent graft versus-host disease (GVHD) is limited in pediatric patients undergoing allogeneic blood or bone marrow transplants. We reviewed our use of this regimen in 24 pediatric patients who had 26 blood or marrow transplants. Acute GVHD occurred in 7 patients (4 unrelated donor transplants, 3 matched sibling transplants; 5 grade I to II, 1 grade III, and 1 not classifiable). One patient had extensive chronic GVHD (matched sibling transplant). In our experience, tacrolimus with mini methotrexate has been well tolerated with minimal toxicity. PMID- 19131789 TI - Early recognition of renal toxicity of high-dose methotrexate therapy: a case report. AB - A 10-year-old boy with osteosarcoma and normal renal function manifested laboratory evidence of impending renal toxicity and extreme elevation of aspartate aminotrasferase and alanine aminotransferase within 2 hours after the completion of a 4-hour infusion of high-dose methotrexate (MTX) (12 g/m2), and went on to develop acute renal failure with life-threatening hyperkalemia 29 hours later. Although his renal function recovered completely with high-dose leucovorin, hemodialysis, charcoal hemoperfusion, and carboxypeptidase G2, we present this case to emphasize that signs of renal toxicity may be present as early as 2 hours after the completion of a 4-hour MTX infusion, and to suggest that monitoring for MTX toxicity should perhaps begin within a few hours after the completion of 4-hour MTX infusion. PMID- 19131790 TI - Diagnostic value of platelet indices in children with leukemia. AB - The purpose of this study was to investigate whether platelet indices [mean platelet volume (MPV) and platelet distribution width (PDW)] could serve as diagnostic tools for screening or as surrogate marker for follow-up in children with leukemia. Blood samples were obtained from 47 patients with leukemia at diagnosis before chemotherapy (mean age: 67 mo; 30 males and 17 females) and from 47 healthy controls (mean age: 59 mo; 27 males and 20 females). We measured the blood platelet indices using an automated counter. MPV was higher in the leukemia group, but it was not statistically significant. However, PDW was significantly lower (P<0.001) in leukemia group. There was no significant difference in the MPV or PDW in patients with acute lymphoblastic leukemia versus acute myeloblastic leukemia. In conclusion, we found no significant difference in the MPV between the 2 groups. PDW proposed as indicators of certain pathologic conditions and it seems possible to use PDW as screening. However, platelet indices (MPV and PDW) cannot be used as indicator to discriminate between the subtypes of leukemia in children. The potential role of platelet indices in leukemia remains to be investigated by a multi-institutional level to verify the possible clinical significance of this finding. PMID- 19131791 TI - Hemophagocytic lymphohistiocytosis onset during induction therapy for precursor B cell acute lymphoblastic leukemia. AB - We report the clinical course of a child with precursor B-cell acute lymphoblastic leukemia who developed fever, hepatosplenomegaly, and refractory thrombocytopenia after initiation of chemotherapy. Bone marrow aspiration on Day 8 of induction showed new onset of hemophagocytosis and he met clinical, laboratory, and histopathologic criteria for hemophagocytic lymphohistiocytosis. His symptoms improved significantly after an increased dosage of steroids. PMID- 19131792 TI - Refractory nodular lymphocyte predominant Hodgkin lymphoma transformed to T cell/histiocyte-rich B-cell lymphoma in an adolescent: salvage therapy with allogeneic bone marrow transplantation. AB - According to biologic features, there is a substantial "gray zone" between nodular lymphocyte predominant Hodgkin lymphomas (NLPHLs) (Poppema lymphomas) and T-cell/histiocyte-rich B-cell lymphomas (T/HRBCLs). Transformation from an NLPHL to a T/HRBCL can occur and is associated with a worsening of the prognosis. Here is described a case of a 16-year-old boy who presented with an NLPHL with features of T/HRBCL. Clinical evolution was complicated by 2 relapses leading to autologous and then to allogeneic bone marrow transplantation. PMID- 19131793 TI - Primary leptomeningeal ALK+ lymphoma in a 13-year-old child. AB - A distinct pathologic entity characterized by expression of the anaplastic lymphoma kinase (ALK) protein (hence described as ALK lymphoma) has emerged within the heterogeneous group of CD30 anaplastic large-cell lymphomas. Central nervous system (CNS) involvement is extremely rare in anaplastic large-cell lymphoma. In children, only isolated cases have been reported, mainly as secondary CNS involvement. We report on a 13-year-old boy presenting with headaches and diplopia. Cerebrospinal fluid was infiltrated with atypical large granular lymphocytes. Magnetic resonance imaging of the brain revealed leptomeningeal enhancement. A frontal lobe biopsy showed a pleomorphic neoplasm diffusely infiltrating the meninges composed of large cells with bizarre nuclei similar to those evidenced in cerebrospinal fluid. Immunohistochemical stains showed diffuse strong positivity for CD8, CD30, anaplastic lymphoma kinase protein: p80 and negative monocyte-macrophage and B cell markers. TCR gamma was clonally rearranged. This finding was confirmed by reverse transcription polymerase chain reaction analysis of the NPM/ALK fusion protein. Epstein-Barr virus was not detected. No evidence of extra-CNS disease was found by imaging study, cytologic examination, or molecular studies. The patient underwent complete remission with polychemotherapy followed by a CNS irradiation. At +10 months from onset, he suffered a full relapse. After a short-term remission with vinblastine, he underwent nonmyeloablative allogeneic bone marrow transplantation, but unfortunately died from multiple organ failure. This case is the first reported occurrence of a primary meningeal ALK lymphoma in a child. PMID- 19131794 TI - Fibrolamellar hepatocellular carcinoma in an infant and literature review. AB - Hepatocellular carcinoma (HCC) is a rare pediatric neoplasm exceptionally reported in infants and fibrolamellar hepatocarcinoma (FLC) a HCC variant. Controversy exists whether FLC has a better prognosis than classic HCC, although recent studies of children and young adults with FLC did not report a better outcome. We present a 4-month-old male infant without any related metabolic or infectious disease who developed a metastatic and multifocal FLC. Serum alpha fetoprotein determinations were always normal. Induction chemotherapy using cisplatin and Adriamycin resulted in a partial response, however, refractory disease developed and regional metastasis precluded surgical resection. The child died from tumoral progression. PMID- 19131795 TI - Successful salvage chemotherapy for isolated central nervous system (CNS) relapse in Burkitt lymphoma: monocentric experience of 3 pediatric patients. AB - Short-term intensive chemotherapy regimens have substantially improved the prognosis of pediatric patients with Burkitt lymphoma (BL), which now has an excellent overall outcome. However, central nervous system (CNS) involvement at diagnosis remains a poor prognostic factor, and progressive or relapsed disease in the CNS is associated with even worse outcomes. We report 3 boys aged 4, 7, and 12 years treated under the French Societe Francaise d'Oncologie Pediatrique LMB 89/96 protocols who presented, respectively, with CNS-/bone marrow+ stage-IV BL; CNS+ stage-IV BL; and stage-I BL. Each experienced an isolated CNS relapse, which was treated with CNS-directed salvage chemotherapy. All 3 are alive after 11 years of median follow-up, indicating that this chemotherapy regimen can be curative in pediatric BL with isolated CNS relapse. PMID- 19131796 TI - Vanishing bile duct syndrome and Hodgkin disease: a case series and review of the literature. AB - This is the first description in which the diagnosis of vanishing bile duct syndrome (VBDS) preceded the diagnosis of Hodgkin disease (HD) by several months, and for which patients received modifications to modern MOPP-ABV chemotherapy with successful clinical remission. VBDS is an uncommon form of liver disease manifested by severe cholestasis and progressive liver failure. We report 2 cases of stage IIIB pediatric HD and VBDS. Because VBDS is progressive and the only curative treatment is liver transplant, it is imperative to recognize that children with VBDS may also have concurrent HD. PMID- 19131797 TI - Granulocytic sarcoma of the colon in a child with acute myeloid leukemia presenting as hematochezia. AB - Granulocytic sarcoma (GS), an extramedullary myeloid tumor composed of immature cells of the granulocytic series, can occur in patients with acute myeloid leukemia (AML), myelodysplastic syndrome, or chronic myelogenous leukemia. It can occur in any organ or tissue, but the most common involved areas are the skin, bone/spine, and lymph nodes. However, its occurrence in the gastrointestinal tract is relatively rare, and is especially rare in the colon in adults. No case of GS involving the colon in children has ever been reported. We report here an extremely rare case of GS in the colon of a 10-year-old boy with AML presenting with hematochezia. Colonic GS was diagnosed by colonofiberscopic biopsy. His hematochezia responded rapidly to induction chemotherapy and the patient remained in complete remission after 3-month follow-up. In conclusion, hematochezia may be due to colonic involvement of GS, which should be considered in the differentials in addition to thrombocytopenia, as it is usually encountered in AML patients. PMID- 19131798 TI - Pediatric gastric lymphoma: a rare entity. AB - Primary gastric lymphoma is a rare event in childhood. We describe a 13-year-old boy with gastric Burkitt-like lymphoma localized in the fundus. Symptoms mimicking gastritis-epigastric pain, hypochromic anemia, anorexia, and weight loss had been present for a few months before diagnosis. No Helicobacter pylori infection was shown at diagnosis. Biopsies obtained by ultrasound gastroscopy proved the diagnosis; F-fluorodeoxyglucose-positron emission tomography detected an isolated large gastric hypermetabolic mass. According to the international FAB/LMB 96 trial, the patient was treated with chemotherapy alone and is in first complete remission 2(1/2) years after diagnosis. PMID- 19131799 TI - Black tumor, an unusual yolk sac tumor with massive melanin deposition. PMID- 19131800 TI - The American Association for the Surgery of Trauma--through the looking glass: deja vu all over again. PMID- 19131802 TI - The utility of early end-tidal capnography in monitoring ventilation status after severe injury. AB - BACKGROUND: An arterial CO2 (Paco2) of 30 mm Hg to 39 mm Hg has been shown to be the ideal target range for early ventilation in trauma patients; however, this requires serial arterial blood gases. The use of end-tidal capnography (EtCO2) has been recommended as a surrogate measure of ventilation in the prehospital arena. This is based on the observation of close EtCO2 Pa(CO2) correlation in healthy patients, yet trauma patients frequently suffer from impaired pulmonary ventilation/perfusion. Thus, we hypothesize that EtCO2 will demonstrate a poor reflection of actual ventilation status after severe injury. METHODS: Prospective observational study on consecutive intubated trauma patients treated in our emergency department (ED) during 9 months. Arterial blood gas values and concomitant EtCO2 levels were recorded. Regression was used to determine the strength of correlation among all trauma patients and subgroups based on injury severity (Abbreviated Injury Score and Injury Severity Score) and physiologic markers of perfusion status (lactate, shock index, and arterial base deficit). RESULTS: During 9 months, 180 patients were evaluated. The EtCO2 Paco2 correlation was poor at R2 = 0.277. Patients ventilated in the recommended EtCO2 (range, 35 to 40) were likely to be under ventilated (Pa(CO2) > 40 mm Hg) 80% of the time, and severely under ventilated (Pa(CO2) > 50 mm Hg) 30% of the time. Correlation was best for patients with isolated traumatic brain injury and worst for those with evidence of poor tissue perfusion. CONCLUSION: EtCO2 has low correlation with Pa(CO2), and therefore should not be used to guide ventilation in intubated trauma patients in the ED. Better strategies for guiding prehospital and ED ventilation are needed. PMID- 19131801 TI - Pentoxifylline modulates intestinal tight junction signaling after burn injury: effects on myosin light chain kinase. AB - BACKGROUND: Burn injury can result in loss of intestinal barrier function, leading to systemic inflammatory response syndrome and multiorgan failure. Myosin light chain kinase (MLCK), a tight junction protein involved in the regulation of barrier function, increases intestinal epithelial permeability when activated. Prior studies have shown that tumor necrosis factor (TNF)-alpha activates MLCK, in part through a nuclear factor (NF)-kappa B-dependent pathway. We have previously shown that pentoxifylline (PTX) decreases both TNF-alpha synthesis and NF-kappaB activation in models of shock. Therefore, we postulate that PTX will attenuate activation of the tight junction protein MLCK, which may decrease intestinal tight junction permeability after severe burn. METHODS: Male balb/c mice undergoing a severe burn were randomized to resuscitation with normal saline (NS) or NS + PTX (12.5 mg/kg). Intestinal TNF-alpha levels were evaluated using enzyme linked immunosorbent assay. Gut extracts were obtained to assess MLCK, phosphorylated IKK, IkappaB-alpha, and NF-kappaB p65 levels by immunoblotting. RESULTS: Burn injury increased intestinal MLCK protein levels threefold in animals resuscitated with NS, whereas those receiving PTX had MLCK levels similar to control (p < 0.01). Treatment with PTX attenuated burn-induced intestinal permeability. PTX decreased cytoplasmic IKK, IkappaB-alpha phosphorylation, and nuclear NF-kappaB p65 translocation to sham levels (p < 0.05 vs. NS). CONCLUSION: Treatment with PTX attenuates activation of the tight junction protein MLCK, likely through its ability to decrease local TNF-alpha synthesis and NF-kappaB activation after burn. PTX may have therapeutic utility by decreasing intestinal barrier breakdown after burn. PMID- 19131803 TI - Early intubation in the management of trauma patients: indications and outcomes in 1,000 consecutive patients. AB - BACKGROUND: The Eastern Association for the Surgery of Trauma Practice Management Guidelines identify indications (EI) for early intubation. However, EI have not been clinically validated. Many intubations are performed for other discretionary indications (DI). We evaluated early intubation to assess the incidence and outcomes of those performed for both EI and DI. METHODS: One thousand consecutive intubations performed in the first 2 hours after arrival at our Level I trauma center were reviewed. Indications, outcomes, and trauma surgeon (TS) intubation rates were evaluated. RESULTS: During a 56-month period, 1,000 (9.9%) of 10,137 trauma patients were intubated within 2 hours of arrival. DI were present in 444 (44.4%) and EI in 556 (55.6%). DI were combativeness or altered mental status in 375 (84.5%), airway or respiratory problems in 21 (4.7%), and preoperative management in 48 (10.8%). Injury Severity Score was 14.6 in DI patients and 22.7 in EI patients (p < 0.001). Predicted versus observed survival was 96.6% versus 95.9% in DI patients and 75.2% versus 75.0% in EI patients (p < 0.001). Head Abbreviated Injury Scale score of >or=3 occurred in 32.7% with DI and 52.0% with EI (p < 0.001). Seven (0.7%) surgical airways were performed; two for DI (0.2%). Eleven (1.1%) patients aspirated during intubation and five (0.5%) suffered oral trauma. There were no other significant complications of intubation for either DI or EI and complication rates were similar in the two groups. Delayed intubation (early intubation after leaving the trauma bay) was required in 67 (6.7%) patients and 59 (88.1%) were for combativeness, neurologic deterioration, or respiratory distress or airway problems. Intubation rates varied among TS from 7.6% to 15.3% (p < 0.001) and rates for DI ranged from 3.3% to 7.4% (p < 0.001). There was a statistically insignificant trend among TS with higher intubation rates to perform fewer delayed intubations. CONCLUSIONS: Early intubation for EI as well as DI was safe and effective. One third of the DI patients had significant head injury. Surgical airways were rarely needed and delayed intubations were uncommon. The intubation rates for EI and DI varied significantly among TSs. The Eastern Association for the Surgery of Trauma Guidelines may not identify all patients who would benefit from early intubation after injury. PMID- 19131804 TI - Predefined massive transfusion protocols are associated with a reduction in organ failure and postinjury complications. AB - INTRODUCTION: Massive transfusion (MT) protocols have been shown to improve survival in severely injured patients. However, others have noted that these higher fresh frozen plasma (FFP):red blood cell (RBC) ratios are associated with increased risk of organ failure. The purpose of this study was to determine whether MT protocols are associated with increased organ failure and complications. METHODS: Our institution's exsanguination protocol (TEP) involves the immediate delivery of products in a 3:2 ratio of RBC:FFP and 5:1 for RBC:platelets. All patients receiving TEP between February 2006 and January 2008 were compared with a cohort (pre-TEP) of all patients from February 2004 to January 2006 that (1) went immediately to the operating room and (2) received MT (>or=10 units of RBC in first 24 hours). RESULTS: Two hundred sixty-four patients met inclusion (125 in the TEP group, 141 in the pre-TEP). Demographics and Injury Severity Score were similar. TEP received more intraoperative FFP and platelets but less in first 24 hours (p < 0.01). There was no difference in renal failure or systemic inflammatory response syndrome, but pneumonia, pulmonary failure, open abdomens, and abdominal compartment syndrome were lower in TEP. In addition, severe sepsis or septic shock and multiorgan failure were both lower in the TEP patients (9% vs. 20%, p = 0.011 and 16% vs. 37%, p < 0.001, respectively). CONCLUSIONS: Although MT has been associated with higher organ failure and complication rates, this risk appears to be reduced when blood products are delivered early in the resuscitation through a predefined protocol. Our institution's TEP was associated with a reduction in multiorgan failure and infectious complications, as well as an increase in ventilator-free days. In addition, implementation of this protocol was followed by a dramatic reduction in development of abdominal compartment syndrome and the incidence of open abdomens. PMID- 19131805 TI - Sleep deprivation after septic insult increases mortality independent of age. AB - BACKGROUND: Sleep deprivation is a common problem in the intensive care unit. Animal models have demonstrated that sleep deprivation alone is associated with increased mortality. We have previously shown that septic insult with sleep deprivation results in increased mortality in a murine model. The aging process is known to reduce the restorative phases of sleep. The purpose of this study was to evaluate the effect of age on mortality with sleep deprivation during recovery from septic insult. METHODS: C57BL/6J male mice aged 2 months (young) or 9 months (old) underwent cecal ligation and puncture (CLP). Animals were randomized to receive sleep interruption (SI) for 48 hours or standard recovery (no SI). Sham animals underwent laparotomy and cecal manipulation without puncture. SI was achieved by securing animal housing to an orbital shaker set to repeatedly cycle at 30 rpm over 120 seconds (30 seconds on/90 seconds off). The primary outcome was survival at 5 days post-CLP. Kaplan-Meier survival analysis with log-rank test was used to explore differences in mortality. RESULTS: SI resulted in an increase in time awake for both light and dark cycles (p < 0.001). Mortality after CLP with SI (n = 30) was 57% and mortality after CLP without SI (controls; n = 33) was 24%. SI was associated with a greater than 3-fold increase in mortality after CLP (RR = 3.29; 95% CI, 1.42-7.63). Young mice (n = 28) had a mortality of 31% with CLP alone increasing to 67% with SI (p = 0.03). Old mice (n = 35) had a mortality of 18% with CLP alone increasing to 50% with SI (p = 0.05). There was no difference in survival between young and old mice undergoing SI (p = 0.49). CONCLUSIONS: Sleep deprivation after septic insult increases mortality in both young and old mice. However, sleep deprivation after septic insult does not have a more profound effect on mortality in either age group. These findings suggest that sleep deprivation experienced in the intensive care unit setting during recovery from critical illness may increase mortality. This effect appears independent of increased age. Further studies evaluating extremes of age are warranted. PMID- 19131806 TI - Coagulopathy in severe traumatic brain injury: a prospective study. AB - BACKGROUND: The incidence and risk factors for traumatic brain injury (TBI) associated coagulopathy after severe TBI (sTBI) and the effect of this complication on outcomes have not been evaluated in any large prospective studies. METHODS: Prospective study of all patients admitted to the surgical intensive care unit (ICU) of an urban, Level I trauma center from June 2005 through May 2007 with sTBI (head Abbreviated Injury Scale score of >or=3). Criteria for TBI-coagulopathy included a clinical condition consistent with coagulopathy, i.e. sTBI, in conjunction with a platelet count <100,000 mm3 and/or elevated international normalized ratio and/or activated partial thromboplastin time. The following potential risk factors with p < 0.2 on bivariate analysis were included in a stepwise logistic regression analysis to identify independent risk factors for TBI coagulopathy and its association with mortality: age, mechanism of injury (blunt [B] or penetrating [P]), presence of hypotension upon admission, Injury Severity Score (ISS), Glasgow Coma Scale (GCS), head and other body area Abbreviated Injury Scale, isolated head injury, diffuse axonal injury, cerebral edema, intracranial hemorrhage (intraventricular, parenchymal, subarachnoid, or subdural), pneumocephalus, and presence of midline shift. RESULTS: A total of 436 patients (392 blunt, 44 penetrating) met study criteria, of whom 387 patients had isolated SHI. TBI coagulopathy occurred in 36% of all patients (B: 33%, P: 55%; p < 0.0075) and in 34% of patients with isolated head injury (B: 32%, P: 54%; p = 0.0062). Independent risk factors for TBI coagulopathy in isolated sTBI were found to include a GCS score of or=16, presence of cerebral edema, subarachnoid hemorrhage, and midline shift. ICU lengths of stay were significantly longer in SHI patients who developed TBI coagulopathy (12.7 vs. 8.8 days; p = 0.006). The development of TBI coagulopathy in SHI was associated with increased mortality, adjusted odds ratio (95% confidence interval): 9.61 (4.06-25.0); p < 0.0001. CONCLUSION: The incidence of TBI coagulopathy in SHI is high, especially in penetrating injuries. Independent risk factors for coagulopathy in isolated head injuries include GCS score of or=16, hypotension upon admission, cerebral edema, subarachnoid hemorrhage, and midline shift. The development of TBI coagulopathy is associated with longer ICU length of stay and an almost 10-fold increased risk of death. PMID- 19131807 TI - Reversal of coagulopathy in critically ill patients with traumatic brain injury: recombinant factor VIIa is more cost-effective than plasma. AB - BACKGROUND: Traumatic brain injury (TBI) is the leading cause of death and disability after trauma. Coagulopathy is common in this patient population and requires rapid reversal to allow for safe neurosurgical intervention and prevent worsening of the primary injury. Typically reversal of coagulopathy is accomplished with the use of plasma. Recombinant factor VIIa (rFVIIa; NovoSeven, Novo Nordisk, Bagsvaerd, Denmark) has become increasingly used "off-label" in patients with neurosurgical emergencies to rapidly reverse coagulopathy. We hypothesized that the use of rFVIIa in this patient population would prove to be cost-effective as well as demonstrate clinical benefit. METHODS: The trauma registry at the R Adams Cowley Shock Trauma Center was used to identify all coagulopatic trauma patients admitted between January 2002 and December 2007 with relatively isolated TBI (head Abbreviated Injury Scale score of >or=4). The medical records of patients were reviewed and demographics, injury-specific data, medications administered, laboratory values, blood product utilization, neurosurgical procedures, length of stay (LOS), discharge disposition, and outcome data were abstracted. Patients who received rFVIIa for reversal of coagulopathy were compared against those who did not receive rFVIIa. t Tests were used to compare differences between continuous variables, and chi2 analysis was used to compare categorical variables. A p value of <0.05 was considered significant for all statistical tests. RESULTS: During a 6-year period, there were 179 patients who met inclusion criteria. One hundred eleven patients (62.0%) were treated with conventional therapy alone whereas 68 (38.0%) received rFVIIa. Baseline characteristics between the two groups were similar except that Injury Severity Score and admission International normalized ratio were higher in the rFVIIa group and the rFVIIa group had a higher percentage of patients with head Abbreviated Injury Scale score of 5 injuries, patients who underwent neurosurgical procedures and patients with preinjury warfarin use. There was no difference in total charges between these groups (mean US $63,403 in the conventionally treated group vs. $66,086). When patients who required admission to the intensive care unit were analyzed (n = 110, 50% received rFVIIa), total mean charges and costs were significantly lower in the group that received rFVIIa (mean US $108,900 vs. $77,907). Hospital LOS, days of mechanical ventilation, and plasma utilization were lower in the rFVIIa group. Mortality and thromboembolic complication rates were not different between the two groups. CONCLUSION: In this study, we were able to demonstrate a significant economic benefit of the use of rFVIIa for reversal of coagulopathy in severely injured patients with TBI. Not all patients with coagulopathy and an anatomic brain injury benefit, but in patients who are neurologically or physiologically compromised, using rFVIIa decreases total charges and costs of hospitalization. This decrease in overall cost is directly attributable to the significant decrease in LOS and decrease in the need for mechanical ventilation. This study demonstrates that in coagulopathic patients with TBI who require intensive care unit admission, rFVIIa is cost-effective and safe. Prospective studies are needed to confirm these findings and establish clinical effectiveness. PMID- 19131808 TI - The effect of storage on the accumulation of oxidative biomarkers in donated packed red blood cells. AB - BACKGROUND: Transfusion-related acute lung injury (TRALI) is a life-threatening condition characterized by oxidative stress. Longer storage times of packed red blood cells (PRBC) and other blood products have been implicated with an increased risk in developing TRALI in transfused patients. METHODS: A total of 10 units of blood containing PRBC stored in citrate-phosphate-dextrose buffer at 4 degrees C were included in the study. At Bonfils Blood Center (Denver, CO), samples were collected on storage day 1 and day 42. Samples were immediately centrifuged, and the supernatants were collected and stored at -80 degrees C until further analysis. Oxidation-reduction potential and protein oxidation were measured in both the day 1 and day 42 samples. RESULTS: Oxidation-reduction potential significantly increased (p < 0.05) in the day 42 sample (98.1 mV +/- 21.9 SD) versus the day 1 sample (62.6 mV +/- 21.5 SD). The oxidation of human serum albumin increased by 63.6% during the storage time. Other serum proteins such as apolipoprotein A1 and transthyretin demonstrated similar increases in oxidation. Also, proteins with a cleaved C-terminal amino acid were observed indicating the presence of carboxypeptidase activity, a marker of inflammation. CONCLUSIONS: The presence of an oxidative environment in transfused PRBC increases with storage time. This could partially explain the increased risk of developing TRALI related to the transfusion of older blood products. PMID- 19131809 TI - Mechanisms of delayed wound healing by commonly used antiseptics. AB - BACKGROUND: The cytotoxic effects of antiseptics on pivotal cell types of the healing process have been well documented. The purpose of our investigation was to explore the ability of subcytotoxic levels of antiseptics to interfere with fibroblast function. METHODS: Cell proliferation assays were performed by culturing fibroblasts in the presence of commonly used antiseptics. Migration was evaluated using scratch assays in which monolayers were "wounded" and cellular movement was monitored by digital photography. Matrix metalloproteinase (MMP) release was analyzed by zymography. RESULTS: H2O2 and povidone-iodine reduced both migration and proliferation of fibroblasts in a dose-dependent fashion. Treatment with silver-containing antiseptics and chlorhexidine exhibited reductions in proliferation at high concentrations, but enhanced growth at lower doses. Silver-containing compounds and chlorhexidine also proved to be the least detrimental to migration in these assays. metalloproteinase release from the cells was differently affected depending on the dosage and class of antiseptic applied. CONCLUSIONS: When debridement of the wound bed is not sufficient to reduce bacterial loads, the application of broad-spectrum antiseptics maybe indicated. Our data would suggest that H2O2 and iodine are poor choices, potentially retarding the contribution of fibroblasts to the healing process. Silver sulfadiazine and chlorhexidine, at levels still proven to be bactericidal, had fewer detrimental effects on fibroblast activity in these assays. The silver containing antiseptics may even increase the proliferative potential of these cells in culture. PMID- 19131810 TI - 34th William Fitts Jr oration: the parathyroid response to acute hemorrhage, sepsis, and multiple organ failure. PMID- 19131811 TI - The standardized live patient and mechanical patient models--their roles in trauma teaching. AB - BACKGROUND: We have previously demonstrated improved medical student performance using standardized live patient models in the Trauma Evaluation and Management (TEAM) program. The trauma manikin has also been offered as an option for teaching trauma skills in this program. In this study, we compare performance using both models. METHODS: Final year medical students were randomly assigned to three groups: group I (n = 22) with neither model, group II (n = 24) with patient model, and group III (n = 24) with mechanical model using the same clinical scenario. All students completed pre-TEAM and post-TEAM multiple choice question (MCQ) exams and an evaluation questionnaire scoring five items on a scale of 1 to 5 with 5 being the highest. The items were objectives were met, knowledge improved, skills improved, overall satisfaction, and course should be mandatory. Students (groups II and III) then switched models, rating preferences in six categories: more challenging, more interesting, more dynamic, more enjoyable learning, more realistic, and overall better model. Scores were analyzed by ANOVA with p < 0.05 being considered statistically significant. RESULTS: All groups had similar scores (means % +/- SD)in the pretest (group I - 50.8 +/- 7.4, group II - 51.3 +/- 6.4, group III - 51.1 +/- 6.6). All groups improved their post-test scores but groups II and III scored higher than group I with no difference in scores between groups II and III (group I - 77.5 +/- 3.8, group II - 84.8 +/- 3.6, group III - 86.3 +/- 3.2). The percent of students scoring 5 in the questionnaire are as follows: objectives met - 100% for all groups; knowledge improved: group I - 91%, group II - 96%, group III - 92%; skills improved: group I - 9%, group II - 83%, group III - 96%; overall satisfaction: group I - 91%, group II - 92%, group III - 92%; should be mandatory: group I - 32%, group II - 96%, group III - 100%. Student preferences (48 students) are as follows: the mechanical model was more challenging (44 of 48); more interesting (40 of 48); more dynamic (46 of 48); more enjoyable (48 of 48); more realistic (32/48), and better overall model (42 of 48). CONCLUSIONS: Using the TEAM program, we have demonstrated that improvement in knowledge and skills are equally enhanced by using mechanical or patient models in trauma teaching. However, students overwhelmingly preferred the mechanical model. PMID- 19131812 TI - Bioprosthetic repair of complex duodenal injury in a porcine model. AB - BACKGROUND: Complex duodenal injury remains a challenging problem for the trauma surgeon. Although primary repair of small injuries is often possible, extensive damage requires complex enteric reconstruction and drainage procedures. We sought to determine the efficacy of a bioprosthetic repair for large duodenal wounds in a porcine model. METHODS: A 60% circumferential wall defect in the second portion of the duodenum was created in eight female Yorkshire swine (38 kg +/- 5 kg). After 30 minutes of peritoneal soilage, a bioprosthetic repair using 1.5 mm porcine acellular dermal matrix was performed. Animals were recovered and resumed a normal diet on day 3. Repeat abdominal exploration and anastomotic bursting pressure strength was performed at 1-, 2-, 3-, and 6-week intervals. Pathologic analysis of all specimens was performed. RESULTS: All animals tolerated a normal diet postoperatively, with progressive weight gain and normal bowel function. On re-exploration, no animal had evidence of duodenal stenosis, proximal dilation, or abscess formation. Pathologic analysis demonstrated progressive in-growth of native bowel tissue, with almost complete incorporation at 6 weeks. Mean bursting pressure (202 mm Hg +/- 60 mm Hg) occurred at native bowel, not patch repair site, in three of eight animals. CONCLUSION: Bioprosthetic repair of enteric wall defects, even in proximity to upper intestinal secretions, allows successful recovery of bowel function and injury repair without extensive anatomic reconstruction. This technique may provide a more conservative approach to the treatment of complex duodenal injuries after trauma. PMID- 19131813 TI - Enteral supplementation enriched with glutamine, fiber, and oligosaccharide prevents gut translocation in a bacterial overgrowth model. AB - BACKGROUND: Normal gut flora plays an important role in the intestinal mucosal barrier function under various critical conditions. The flora may alter after severe insults, such as trauma and shock. Enteral nutrition should preserve the gut environment; however, full support is usually difficult for severely ill patients because of impaired gastrointestinal motility. Currently, we have commercial enteral supplementation product enriched with glutamine, dietary fiber, and oligosaccharide (GFO) in Japan. This study examines the hypothesis that the enteral supplementation ameliorates gut injury induced by a bacterial overgrowth model, even in small volumes and quantities. MATERIALS: Balb/c mice received antibiotics (4 mg/mL of streptomycin) in their drinking water for 4 days to kill the normal gut flora after which they were orally inoculated with a streptomycin-resistant strain of Escherichia coli, known as E. coli C-25. The mice that were administered bacterial monoassociation received 0.5 mL of GFO twice daily (GFO group) or 10% of glucose solution (GLU group). Unsupplemented drinking water was used for control animals (control) whose gut flora was normal. The mice were killed and their mesenteric lymph nodes complex was harvested and processed to test gut bacterial translocation. The cecal population levels of bacteria and ileum histology were also examined. RESULTS: The incidence and magnitude of gut translocation to the lymph nodes complex in the GLU group were significantly higher than those in the control (p < 0.01). Treatment with GFO prevented the gut translocation although animals in the GFO group had same level of the cecal bacterial population. Histologic findings in the ileum were not different between the GLU and GFO. CONCLUSION: GFOs supplement prevented gut translocation for bacterial overgrowth even in small volumes and quantities. The intestinal histologic findings could not explain the protective mechanisms of GFO. Further studies may be needed to elucidate the benefit of the partial enteral nutrition. PMID- 19131814 TI - Variation in the TLR4 gene influences the risk of organ failure and shock posttrauma: a cohort study. AB - BACKGROUND: Genetic variation contributes to risk and outcomes of sepsis. We sought to determine whether variation in inflammation related genes is associated with severity of sepsis in trauma patients. METHODS: A cohort of severely injured Caucasian patients was studied and genotyped for candidate single nucleotide polymorphisms (SNPs). These were toll-like receptor 4 (TLR4) A896G, tumor necrosis factor-alpha G-308A, interleukin-6 G-174C, interleukin-1beta C-31T, and cluster of differentiation marker 14C-159T. SNP genotypes among patients with sepsis and complicated sepsis were analyzed by chi2 and logistic regression. Six haplotype-tagging SNPs in the TLR4 gene were subsequently examined to analyze their influence on TLR4 A896G SNPs relationship to sepsis severity. RESULTS: We enrolled 598 patients. Complicated sepsis developed in 147 (25%). Adjusting for independent risk factors, carriage of the variant TLR4 896 G allele was associated with decreased risk of complicated sepsis (odds ratio = 0.3, 95% confidence interval, 0.1-0.7, p = 0.008). Furthermore, two haplotypes seemed to better characterize this risk than the variant TLR4 896G allele. The variant TLR4 896G allele is linked to one common haplotype, which seems to confer a considerably reduced risk of complicated sepsis. (aOR = 0.2 95% confidence interval, 0.05-0.7, p = 0.01). CONCLUSIONS: Variation within TLR4 gene is associated with severity of posttraumatic sepsis. This risk may not be solely related to TLR4 A896G SNP. It is likely that other, uncharacterized variations in the TLR4 gene contribute to sepsis severity. A thorough evaluation of variability within the TLR4 gene is needed to characterize sepsis risk. PMID- 19131815 TI - TLR4 is essential in acute lung injury induced by unresuscitated hemorrhagic shock. AB - BACKGROUND: Acute lung injury (ALI) and acute respiratory distress syndrome in patients with hemorrhagic shock (HS) or resuscitation is associated with the expression of TLR4. However, the role of TLR4 in ALI induced by unresuscitated HS remains obscure. METHODS: The lung pathologic change was observed by hematoxylin and eosin staining. Interleukin-1beta and tumor necrosis factor-alpha were analyzed by enzyme-linked immunosorbent assay. Polymorphonuclear leukocyte sequestration and lung leak were analyzed by pulmonary myeloperoxidase activity and Evans blue dye. The expressions of TLR4 mRNA and protein were analyzed by reverse transcription-polymerase chain reaction and Western blot, respectively. TLR4 distribution was analyzed by immunohistochemistry. RESULTS: Lung neutrophil accumulation and microvascular permeability were significantly increased after unresuscitated HS, meanwhile, lung interleukin-1beta and tumor necrosis factor alpha were gradually augmented. TLR4 mRNA, TLR4 distribution and TLR4 protein were also significantly increased in TLR4 wt mice, however, no above-mentioned changes appeared in TLR4 mutant mice. CONCLUSIONS: TLR4 is strongly associated with the pathogenesis of ALI induced by unresuscitated HS, which may serve as a useful therapeutic target. PMID- 19131816 TI - Blunt cerebrovascular injuries: does treatment always matter? AB - BACKGROUND: Blunt cerebrovascular injuries (BCVI) have become an increasingly recognized entity. Stroke as a result of these injuries can have devastating consequences. Optimal screening criteria, diagnostic imaging, and therapy for BCVIs have not been elucidated. Our institution began to apply liberal screening criteria using a whole-body scanning protocol with multidetector computed tomographic (WB-MDCT) scans to diagnose these injuries. The purpose of this study is to describe a single institution's large experience in patients with BCVI in an effort to provide insight into the diagnosis and management of these injuries. METHODS: All patients with a BCVI admitted to the R Adams Cowley Shock Trauma Center during a 30-month period were included in this study. Choice of diagnostic evaluation and treatment regimens were at the discretion of the treating attending physician. Review of medical records and all relevant radiographic studies were retrospectively performed for the purposes of this study. RESULTS: During the study period, there were 12,667 patients admitted to the R Adams Cowley Shock Trauma Center. There were 147 patients identified with 200 carotid or vertebral artery injuries. The incidence of BVCI was 1.2%. Mortality was 13%. Anatomic injury risk factors for BCVI (major facial fractures, skull base fractures, cervical spine fractures or spinal cord injury, or traumatic brain injury) were found in only 78%. Major thoracic injury was found in 63% of patients with carotid artery injuries and cervical spine fractures or spinal cord injury was found in 74% of patients with vertebral artery injuries. The initial screening test employed was a WB-MDCT in 96% of patients of which 84% detected a BCVI. Treatments included endovascular therapy (22%), antiplatelet medications (36%), anticoagulation (10%), and combination therapy with antiplatelet agents and anticoagulation (18%). Thirty percent received no therapy, primarily due to contraindications from concomitant injuries. There were 18 (12%) patients who had a stroke. Of these patients, 8 (44%) had evidence of infarction at admission, 6 were diagnosed within 72 hours, and 4 were diagnosed after 1 week. Stroke-related mortality was 50%, whereas clinical follow-up after hospital discharge demonstrated only one patient with disability as a result of infarction. Of 10 patients who did not have stroke at admission, 3 were fully treated, 5 had specific contraindications to therapy, and 2 had no or false-negative imaging before infarction. Stroke rates for untreated patients were 25.8% and patients treated with any therapy had a stroke rate of 3.9% (p = 0.0003). Radiographic follow-up >1 month after injury demonstrated improvement in over 50% of patients. CONCLUSIONS: BCVIs are not infrequent after blunt trauma. These injuries occur even in the absence of classically described risk factors. Liberal screening with WB-MDCT incorporates detection of these injuries into the initial diagnostic evaluation. Stroke occurs in a substantial number of patients and carries a very high mortality. However, nearly one third of patients with BCVI are not candidates for therapy. Treatment does reduce the risk of infarction in patients with BCVI, but strokes, when they occur, are not preventable. PMID- 19131817 TI - Stress insulin resistance is a marker for mortality in traumatic brain injury. AB - BACKGROUND: Both hyper- and hypoglycemia have been associated with poor outcome in traumatic brain injury (TBI). Neither the risks nor benefit of tight glucose control (goal range, 80-110 mg/dL) have been documented in the TBI population. OBJECTIVE: To analyze whether densely collected blood glucose data, using a computerized algorithm, to maintain tight glycemic control will reveal significant differences in blood glucose control between survivors and nonsurvivors in patients with TBI. METHODS: From October 2005 to April 2006, all ventilated, critically ill surgical patients with TBI Abbreviated Injury Scale score of >or=3 were placed on an automated, euglycemia protocol with every 2-hour blood glucose sampling. Mortalities within 24 hours were excluded. The protocol calculates the insulin rate using a linear equation (rate = blood glucose - 60[M]). M is an adapting multiplier and used here as a marker for insulin resistance (IR). RESULTS: Of 1,636 trauma intensive care unit admissions 160 patients, (median Injury Severity Score 34, mortality 13.1%) had 10,071 samples collected. Median glucose 115.6 mg/dL, with 41% of values between 80 and 110 mg/dL, 81% between 80 and 150 mg/dL, and 0.3% <40 mg/dL. The median blood glucose was statistically different but not clinically different among the patients who lived and died (114; interquartile range, 109-132 vs. 118; 111-136, p = 0.01). The median insulin dose was a unit per hour higher among the patient who died (4.2; 2.7-5.9 vs. 3.2; 2.4-5.0, p = 0.006). A logistic regression model demonstrated insulin rate (odds ratio 0.736, 95% confidence interval, 0.549 0.985, p = 0.039) to be the only independent predictor of mortality among the measures of blood glucose control. CONCLUSION: Nonsurvivors with TBI have significantly higher markers of IR (insulin rate and multiplier). Markers of glucose control (median glucose, hypoglycemic episodes, and the percentage of values in range) did not differ clinically among groups. Despite this stress IR, tight glycemic control appears possible and safe with low levels of hypoglycemic episodes in the TBI population. PMID- 19131818 TI - The in vivo effect of propranolol on cerebral perfusion and hypoxia after traumatic brain injury. AB - BACKGROUND: Recent epidemiologic evidence has identified beta-blockade as independently associated with improved survival in patients with isolated traumatic brain injury (TBI). Reduced sympathetic discharge and catecholamine release may improve circulation in the injured areas and influence delayed demise. The purpose of this study was to investigate the cerebral effect of beta blockade in a murine TBI model using immunohistochemical and microPET analysis. METHODS: Balb/c mice underwent TBI as in a previously described model and were randomized to receive treatment with propranolol or placebo in a blinded fashion. Immunofluorescent images were obtained for vessel density (CD31), vessel perfusion (Ricinus communis agglutinin [RCA]-lectin), and cerebral hypoxia (hypoxyprobe-1) and compared by digital quantification. Perfusion measurements were acquired using positron emission tomography microPET scans with [64Cu] pyruvaldehyde bis(N4-methylthiosemicarbazone) ([64Cu]-PTSM) and converted into standardized uptake values (SUV) for analysis. RESULTS: On immunohistochemical analysis, the normal mouse cerebral perfusion was a quantitated mean of 325 +/- 20, the cerebral perfusion after TBI and treatment with placebo was 113 +/- 25, and the cerebral perfusion after TBI treated with propranolol was 172 +/- 23. Immunohistochemical analysis demonstrated treatment with propranolol improved cerebral perfusion by 152% (p value <0.01) and reduced cerebral hypoxia by 24.2% (p value <0.01) compared with treatment with placebo. MicroPET imaging of the normal mouse brain after injection with placebo measured a SUV of 0.7075 +/- 0.02; the normal mouse brain after treatment with propranolol measured a SUV of 0.400 +/- 0.02. After TBI and treatment with placebo, the SUV reduced to 0.395 +/ 0.01; after treatment with propranolol the SUV measured 0.515 +/- 0.04. MicroPET imaging demonstrated propranolol improved cerebral perfusion after TBI to 130% of placebo (p value <0.01). CONCLUSION: Propranolol in vivo increased cerebral perfusion and decreased cerebral hypoxia. This research demonstrates beta blockade may prevent additional brain damage after traumatic insult and should be the focus of future clinical trials. PMID- 19131819 TI - Relationship between maximum intracranial pressure and traumatic lesions detected by T2*-weighted imaging in diffuse axonal injury. AB - BACKGROUND: We prospectively investigated whether the number of traumatic lesions detected by magnetic resonance imaging correlated with the level of intracranial pressure (ICP) and outcome among patients with diffuse axonal injury (DAI). MATERIALS: We prospectively studied 19 patients with closed head injuries who were admitted to the Critical Care and Trauma Services at our medical center and who fulfilled the following criteria: did not recover to a Glasgow Coma Scale score greater than 8 after impact, without fixed dilated pupils, and with initial computed tomography findings compatible with DAI. All subjects (n = 19) underwent ICP monitoring for at least 4 days, and T2*-gradient echo imaging was performed within 4 weeks of the impact. RESULTS: A significantly positive correlation was found between the number of lesions detected by magnetic resonance imaging and maximum ICP. The average Glasgow Coma Scale in the good outcome group (Glasgow Outcome Scale IV and V, n = 8) was significantly greater than that in the poor outcome group (Glasgow Outcome Scale I-III, n = 11). The average maximum ICP and the number of lesions in the good outcome group were significantly less than those in the poor outcome group. CONCLUSION: The number of lesions detected by T2*-gradient echo imaging correlates with the degree of intracranial hypertension and outcome in DAI. PMID- 19131820 TI - Effect of 35 degrees C hypothermia on intracranial pressure and clinical outcome in patients with severe traumatic brain injury. AB - BACKGROUND: From 1994, we have used therapeutic hypothermia in patients with severe traumatic brain injury (Glasgow Coma Scale scores of 5 or less). In 2000, we altered the target temperature to 35 degrees C from the former 33 degrees C, as our findings suggested that cooling to 35 degrees C is sufficient to control intracranial hypertension, and that hypothermia below 35 degrees C may predispose patients to persistent cumulative oxygen debt. We attempted to clarify whether 35 degrees C hypothermia has the same effect as 33 degrees C hypothermia in reducing intracranial hypertension and whether it is associated with fewer complications and improved outcomes. METHODS: We compared intracranial pressure (ICP) and biochemical parameters in the 30 patients treated with 35 degrees C hypothermia (January 2000 to June 2005) with those in the 31 patients treated with 33 degrees C hypothermia (July 1994 to December 1999). RESULTS: Patient characteristics were similar in the two groups. The mean temperature during hypothermia was 35.1 +/- 0.7 degrees C in the 35 degrees C hypothermia group and 33.4 +/- 0.8 degrees C in the 33 degrees C hypothermia group. Mean ICP was controlled under 20 mm Hg during hypothermia in both the 35 degrees C hypothermia and 33 degrees C hypothermia groups. The incidence of intracranial hypertension and low cerebral perfusion pressure did not differ between the two groups. The 35 degrees C hypothermic patients exhibited a significant improvement in the decline of serum potassium concentrations during hypothermia and in the increment of C-reactive protein after rewarming. The mortality rate and the incidence of systemic complications tended to be lower in the 35 degrees C group. CONCLUSIONS: Cooling patients to 35 degrees C is safe and the ICP reduction effects of 35 degrees C hypothermia are similar to those of 33 degrees C hypothermia. PMID- 19131821 TI - Outcome of cervical near-hanging injuries. AB - BACKGROUND: Cervical near-hangings are not rare, but have received little attention in the trauma literature. Increasing numbers of patients received from our local jail and detention centers prompted this study. METHODS: Seventeen-year review of a level I Trauma Center Registry identified 67 patients with cervical strangulation for study. Data were analyzed using the Mann-Whitney test to evaluate continuous predictors, and Fisher's exact test for categorical predictors. RESULTS: Ten of 67 patients died (14.9% mortality). Patients having a lower Glasgow Coma Score (GCS) at the scene (3.5 +/- 1.3 vs. 8.3 +/- 5.0; p = 0.001) and lower GCS in the emergency department (ED) (3.0 +/- 0.0 vs. 9.0 +/- 5.3; p < 0.001) were more likely to die. Injuries consisted predominantly of neck abrasions and anoxic brain injuries (83% mortality). Laryngeal fractures and carotid arterial injuries were detected. No cervical spine fractures were seen, but subluxations were identified. Forty-two percent of the patients were in detention centers when the near-hanging incident occurred. CONCLUSIONS: Cervical near-hangings are referred to the Trauma Service for evaluation. Scene or ED GCS of 3 does not preclude neurologically intact survival, although mortality is high. In our study, the most useful prognostic factors were the need for airway control by intubation or cricothyrotomy, cardiopulmonary resuscitation, lower scene and ED GCS, and cerebral edema on CT Scan. Optimal evaluation includes head and neck CT and CT angiography of the neck. We plan to share these results with local authorities and encourage improvement in risk identification, with earlier involvement of mental health personnel. PMID- 19131822 TI - Trends in hospitalization associated with traumatic brain injury in a publicly insured population, 1992-2002. AB - BACKGROUND: Traumatic brain injury (TBI) is a leading cause of death and disability in developed countries. We document trends in TBI-related hospitalizations in Ontario, Canada, between April 1992 and March 2002, focusing on relationships between inpatient hospitalization rates, age, sex, cause of injury, severity level, and in-hospital mortality. METHODS: Information on all acute hospital separations in Ontario with a diagnosis of TBI was analyzed using logistic regression. RESULTS: Hospitalization rates fell steeply among children and young adults but remained stable among adults aged 66 and older. The proportion of TBI hospitalizations with mild injuries decreased from 75% to 54%, whereas the proportion with moderate injuries increased from 19% to 37%. Adjusting for other risk factors, in-hospital deaths were higher for injuries because of motor vehicle crashes than those because of falls. In-hospital death rates were stable for patients with moderate or severe injuries, but increased over time among those whose injuries were classified as mild, suggesting a trend toward more serious injury within the "mild" classification. CONCLUSIONS: Hospitalizations for TBI involve fewer mild injuries over time and are highest in the oldest segment of the population. PMID- 19131823 TI - Traumatic spinal cord injury mortality, 1981-1998. AB - BACKGROUND: We aim to assess the long-term trend of and identify risk factors for traumatic spinal cord injury (TSCI) mortality from 1981 through 1998 in the state of South Carolina (SC). METHODS: We analyzed data from the TSCI surveillance system in SC. Poisson regression analyses were used to examine trends in TSCI mortality rates across subpopulations of interest. Multiple logistic regression was used to identify risk factors for TSCI mortality. RESULTS: The rate of TSCI mortality was 27.4 per million population between 1981 and 1998. A significant 3% annual decrease in the TSCI mortality rate was found from 1981 through 1998. Specifically, TSCI mortality rates declined the most per year in motor vehicle crashes, males, and whites. Adjusted for covariates, individuals of older ages, black race, with a cervical TSCI, and with a more severe injury, as defined by both Frankel grade and Abbreviated Injury Scale, were associated with higher odds of in-hospital mortality. Females had lower odds of in-hospital mortality than males. CONCLUSION: Although mortality rate is decreasing, TSCI remains a significant public health problem, with SC having higher rates of TSCI mortality than the United States. The association between gender and in-hospital mortality needs further exploration. PMID- 19131824 TI - A biomechanical comparison between the thoracolumbosacral surface contact area (SCA) of a standard backboard with other rigid immobilization surfaces. AB - INTRODUCTION: Backboards are routinely used to protect the spine of trauma patients during transportation. Nevertheless, little is known about the biomechanical properties of this type of immobilization. OBJECTIVES: To evaluate the mechanical support of the thoracolumbosacral spine provided by a standard backboard in comparison with various rigid immobilization surfaces, by examining their respective surface contact area (SCA). MATERIALS: SCAs comparisons of a standard aluminum backboard, a rigid military stretcher, an aluminum backboard covered by blanket, 3 and 5 cm thickness foam, and a cushioned stretcher were made using 12 volunteers. The evaluation was performed by a computer- mediated system that generated a diagram indicating pressure distribution and SCA score in each volunteer. These data were compared with a medical grade mattress, which served as the control group. RESULTS: The median backboard's SCA was 14.6 +/- 5.5 times smaller than the stretcher's SCA (range 4.6-28, average 15, p < 0.001). Its median SCA was essentially doubled by covering it by a standard military blanket and tripled when covered by 3 cm layer of foam. Using a 5-cm layer of foam increased the backboard's SCA by 11 times. Cushioning the stretcher beneath the lumbar spine and the hamstrings by folded blankets, significantly improved its median SCA (96 +/- 31.1, range 36-125, average 89.7). CONCLUSIONS: The backboard's SCA was significantly inferior to all the other surfaces. Although no dynamic evaluation was performed, these data imply that backboards need to be appropriately cushioned or alternate surfaces should be employed to improve the mechanical support during trauma patient transportation. Level of evidence, Level I. PMID- 19131825 TI - Retrograde dynamic locked nailing for femoral supracondylar nonunions after plating. AB - BACKGROUND: No surgical procedure has clearly proven superior in treating femoral supracondylar nonunions after plating. All traditional devices have limitations. A technique for improving the approach of retrograde locked nailing to increase a success rate was developed. METHODS: Twenty-one consecutive adult patients with 21 femoral supracondylar nonunions after plating were treated by lateral approach. Soft-tissue detachment was minimized as much as possible. Retrograde dynamic locked nailing and cancellous bone grafting were consequently performed. Postoperatively, protected weight bearing with exercise of knee range of motion was encouraged as early as possible. RESULTS: Eighteen patients were followed up for an average of 3.3 years (range, 1.2-6.5 years). Sixteen nonunions healed with a union rate of 88.9% and an average union period of 4.2 months (range, 3.5-5.0 months). Although two nonunions persisted without further treatment because of patient hesitation, all patients achieved satisfactory functional outcome. CONCLUSIONS: Although neither surgical technique is clearly superior for treating femoral supracondylar nonunions after plating, the lateral approach with retrograde dynamic locked nailing and cancellous bone grafting may prove to be the optimal treatment alternatives. Minimizing soft-tissue destruction during surgery in addition to protected weight bearing through the treatment course may effectively raise the success rate. PMID- 19131826 TI - Morbidity associated with isolated iliac wing fractures. AB - BACKGROUND: Iliac wing fractures have received little attention in the literature. Commonly occurring after a direct blow, they are rotationally and vertically stable. However, one must be aware of their potential for major morbidity including soft tissue, abdominal organ, and vascular lesions. This study examines the incidence, radiographic findings, soft tissue injuries, and complications associated with isolated iliac wing fractures. METHODS: Retrospective review between 2003 and 2006 of a consecutive series of 450 polytraumatized patients treated in a level-1 University trauma center. Hospital charts and radiographs were reviewed and all patients with an isolated iliac wing fracture were requested to return for a clinical and radiologic examination. RESULTS: One hundred twenty patients had major pelvic trauma. Only 10 sustained a fracture limited to the iliac wing, all after high-energy trauma. Nine of 10 patients sustained a total of 36 additional injuries involving head, thorax, spine, abdomen, urologic, and orthopedic systems. Only two patients underwent internal fixation of their fracture, one because of the fracture compressing the bowel and the other because of a very large rotated iliac wing fragment. Two patients with open fractures were treated with irrigation and debridement and no internal fixation. CONCLUSIONS: An isolated iliac wing fracture not compromising the stability of the pelvic ring may be interpreted as a benign injury. However, serious and potentially life-threatening associated injuries may be present requiring emergency abdominal, vascular, or neurologic surgery. A thorough search for such injuries is critical. Most iliac wing fractures can be managed nonoperatively. PMID- 19131827 TI - Angular stable fixation of proximal humeral fractures. AB - BACKGROUND: Proximal humeral fractures requiring surgical stabilization remain a therapeutic challenge. High rates of treatment-related complications have been reported to be associated, particularly in elderly patients with unstable fracture types and diminished bone quality. In recent years, angular stable implants have been introduced to improve the patients' outcome. The purpose of this study was to assess the clinical and radiographic long-term results after angular stable plate fixation of proximal humeral fractures at this Level I trauma center, with special emphasis on fracture healing, functional outcome and treatment related complications. METHODS: We retrospectively analyzed functional and radiographic results of 42 patients (average age: 57.8 years) after angular stable plate fixation of proximal humeral fractures at an average time of 3.2 (+/ 0.8) years after trauma. Functional results were defined by the Constant shoulder score (CS) and the individual Constant score (CSindiv). Radiographic results were assessed by a three-view trauma series (anteroposterior, lateral, and axillary view). For operative treatment, two different angular stable implants were used. Twenty-seven patients were stabilized by a HOFER plate, fifteen patients by a Locking Proximal Humerus Plate. RESULTS: Clinical results revealed an average CS of 74.0 points and an average individual CSindiv of 79.2% points. Fourteen patients had an excellent functional outcome, eight patients a good outcome and fourteen patients had moderate functional results. Six patients (14%) had a poor outcome with less than 55% on CSindiv. Regarding the radiographic outcome, we had an overall union rate of 95% (40 of 42). Failures of reduction and fixation occurred in fourteen patients (33%), and in nine patients (21%) we had signs of a humeral head necrosis. Re-operation due to treatment-related complications was necessary in two patients (5%). Statistical analysis revealed that the fracture type and the age of the patients had a significant influence on the clinical outcome and on the incidence of treatment-related complications. CONCLUSION: With regards to fracture healing and functional outcome of the patients, we had a satisfactory outcome after angular stable plate fixation of proximal humeral fractures. However, we experienced a notably high rate of technical failures and partial humeral head necrosis. Advanced surgical skills and experiences are necessary to achieve correct anatomic reduction and implant fixation, to reduce the risk of these treatment-related complications. PMID- 19131828 TI - Hemiarthroplasty of the shoulder after four-part fracture of the humeral head: a long-term analysis of 34 cases. AB - OBJECTIVE: To assess the treatment outcomes of patients with four-part fracture of the humeral head after primary and secondary hemiarthroplasty. PATIENTS: Retrospective long-term analysis of 46 patients from 1996 to 2002 of patients with 47 four-part fractures of humeral head. Patients with malignant disease were excluded. INTERVENTION: Aequalis (Tornier, Burscheid, Germany). MAIN OUTCOME MEASUREMENTS: Absolute and relative constant scores at 5-year follow-up examination without age or sex normalization, radiographic parameters of calcification, dislocation of tuberosities, prosthetic loosening, and dislocation of joint. RESULTS: Eighteen patients treated by primary and 16 patients treated by secondary arthroplasty were assessed clinically and radiologically after a mean follow-up of 64 (60-96) months. The absolute Constant scores at follow-up were 54.9 to 48.5 points, respectively. The relative scores were 61.4% and 57.3%, respectively. Dislocation of tuberosities with severe loss of function was found in five cases treated by primary arthroplasty (13.5%) and in 12 treated by secondary arthroplasty (75.0%). CONCLUSIONS: The majority of patients in both groups was free of pain or suffered minor pain as determined by the Constant score. Safe fixation of the tuberosities is a prerequisite for functional exercises and is better achieved in primary arthroplasty. A computed tomography scan before operative therapy aids in making the decision between open reduction and internal fixation or hemiarthroplasty. PMID- 19131829 TI - Scoring mandibular fractures: a tool for staging diagnosis, planning treatment, and predicting prognosis. AB - BACKGROUND: Mandibular fractures (MF) are a daily problem in maxillofacial surgery. A staging system for classifying MF is of paramount importance to plan surgery, to define prognosis, and to exchange information among trauma centers. In this article, a classification for MF is proposed. METHODS: The mandible is divided into six sites (symphysis, body, angle, ramus, coronoid, and condyle) and the mandibular nerve divides the alveolar process (i.e., the upper part containing teeth) from the basal bone (placed caudally with respect to the nerve). MF can be summarized using three abbreviations: A = alveolar, B = basal, and C = complete. Consequently, MFs are staged as follows: F in situ = a greenstick fracture; F1 = a single mobile fragment of alveolar or basal bone; F2 = double mobile fractures of the alveolar or basal portion or a single complete separation of the mandibular arch continuity; F3 = a complete double separation of the mandibular arch continuity; F4 = triple or more mandibular arch fragments. To evaluate the suitability of the proposed classification, a retrospective study on a series of 128 patients (and 205 MFs) was performed. Age, gender, stage, clinical diagnosis at admission, type of surgery, and outcome were considered. RESULTS: A good correlation between the proposed classification and the studied variables was detected. CONCLUSIONS: The new classification is a simple and precise method for staging MF. It can summarize MFs and be used in daily practice. It is our understanding, however, that a multicenter study should be performed before the effectiveness of the proposed classification can be clearly stated. PMID- 19131830 TI - Does acuity matter?--Optimal timing of tracheostomy stratified by injury severity. AB - BACKGROUND: A number of conflicting studies have been conducted to analyze the relationship between the timing of tracheostomy and mortality, intensive care unit (ICU) length of stay (LOS), hospital LOS, and the incidence of pneumonia. In contrast to previous studies, this relationship was investigated in the context of expected survival based on probability of survival (Ps) greater than 25%. METHODS: Trauma patients were screened using a statewide registry during a 5-year period (January 2001 to December 2005). Burn patients, transfer patients, permanent tracheostomies, and patients who underwent multiple surgical airways were excluded from the study. Data were collected on patient demographics, Trauma and Injury Severity Score, days to tracheostomy, mortality, ICU LOS, total ventilator days, pneumonia, and hospital LOS. STATISTICAL ANALYSES: log-linear modeling, chi2, p < 0.05. RESULTS: A total of 125,533 trauma patients were analyzed. Out of these, 82,148 patients met inclusion criteria and had complete data for analysis. There were 6,880 patients intubated at the scene, during transport, or at admission to the emergency department, with 685 receiving a temporary tracheostomy. There was a significantly higher mortality rate (48.9%) associated with patients with low Ps (<0.25) receiving early tracheostomy (ET), <4 days. Among high-Ps patients, the ET group demonstrated reduced ICU LOS, total ventilator days, pneumonia, and hospital LOS (p < 0.05). CONCLUSION: ET in patients with low Ps may not be beneficial given the substantially high mortality rate before post injury day 4. However, ET in high-Ps patients reduces ICU and hospital LOS, total ventilator days, and the incidence of pneumonia. This suggests an increased benefit in ET to trauma patients with high Ps. PMID- 19131831 TI - Morbid obesity is not a risk factor for mortality in critically ill trauma patients. AB - BACKGROUND: Age, Injury severity score (ISS), hyperglycemia (HGL) at admission, and morbid obesity are known risk factors of poor outcome in trauma patients. Our aim was to which risk factors had the highest risk of death in the critically ill trauma patient. METHODS: A Trauma Registry of the American College of Surgeons database retrospective study was performed at our Level I trauma center from January 2000 to October 2004. Inclusion criteria were age >15 years and >or=3 days hospital stay. Data collected included age, gender, and ISS. Groups were divided into nonobese and morbidly obese (MO) (body mass index, BMI >or=40 kg/m2) and into HGL (mean >or=150 mg/dL on initial hospital day) and non-HGL. Primary outcome was 30-day mortality. Differences in mortality and demographic variables between groups were compared using Fisher's exact and Wilcoxon's rank sum tests. Univariate and multivariate logistic regression was used to assess the relationship of HGL, morbid obesity, age, and injury severity to risk of death. Relationships were assessed using odds ratios (OR) and area under the receiver operator characteristic curve (AUC). RESULTS: A total of 1,334 patients met study criteria and 70.5% were male. Demographic means were age 40.3, ISS 25.7, length of stay 13.4, and BMI 27.5. The most common mechanism of injury was motor vehicle collision 55.1%. Overall mortality was 4.7%. Mortality was higher in HGL versus non-HGL (8.7% vs. 3.5%; p < 0.001). Mortality was higher in MO versus nonobese, but not significantly (7.8 vs. 4.6%; not significant [NS] p = 0.222). Univariate logistic regression relationships of death to age OR: 1.031, p < 0.001, AUC +/- SE: 0.639 +/- 0.042; ISS OR: 1.044, p < 0.001, AUC +/- SE: 0.649 +/- 0.039; HGL OR: 2.765, p < 0.001; MO: OR: NS, p = NS, AUC +/- SE: NS. Relationships were similar in a combined multivariate model. CONCLUSION: HGL >150 mg/dL on the day of admission is associated with twofold increase in mortality, and an outcome measure should be followed. Morbid obesity (BMI >or=40) is not an independent risk factor for mortality in the critically ill trauma patient. PMID- 19131832 TI - Intraoperative 3D imaging: value and consequences in 248 cases. AB - BACKGROUND: Intraoperative visualization of articular surfaces is technically demanding, and standard two-dimensional fluoroscopic imaging frequently does not provide adequate detail of nonplanar joints. New imaging modalities allow for intraoperative 3D visualization, which are useful in articular fractures. Purpose of this study was to evaluate the utility of 3D imaging in articular fracture reconstruction. METHODS: In a prospective cohort study, we evaluated 248 consecutive patients with intra-articular fractures. After fracture fixation using standard fluoroscopy, 3D imaging was performed intraoperatively using the Iso-C3D system for all patients. Surgeons filled out questionnaires regarding the utility and perceived accuracy of the 3D system. Postoperative CT scans were performed on approximately half of the patients. Main outcome measurements were based on the surgeons decision to immediately revise the articular reduction or implant position. The setup time for the system was recorded. For patients with postoperative CT scans, articular surface gaps of 2 mm or intra-articular hardware placement was again evaluated. RESULTS: In 19% of all cases, intraoperative image analysis resulted in immediate adjustment of the reduction or hardware exchange. These revisions were based on Iso-C3D views of the articular surface that were not visible using fluoroscopy. Of the 129 postoperative CT scans, five cases revealed a technical error of the joint reconstruction, and a secondary revision procedure was performed. CONCLUSION: In conclusion, the Iso-C3D was a valuable intraoperative tool, providing additional information about the articular surface compared with conventional fluoroscopy in a variety of anatomic regions. PMID- 19131833 TI - The efficacy of magnetic resonance imaging for the diagnosis of testicular rupture: a prospective preliminary study. AB - BACKGROUND: Magnetic resonance imaging (MRI) scan is useful for the evaluation of the testis because of high soft-tissue contrast and its multiplanar capability. The purpose of this study was to evaluate prospectively the efficacy of MRI for the diagnosis of testicular rupture in blunt scrotal trauma. METHODS: From January to December 2006, seven consecutive patients (mean age 22 years; range 21 23 years) that presented with blunt scrotal trauma were enrolled in this prospective study. The patients underwent scrotal sonography (US), and supplementary MRI was performed before exploratory surgery. The essential sequences of the MRI were T2-weighted coronal, axial, and sagittal scans. The diagnostic accuracy of MRI was evaluated by means of the surgical findings as the reference standard. RESULTS: The diagnostic accuracy of MRI was 100% (7 of 7). Three cases with testicular rupture were diagnosed accurately. Interruption of the dark signal intensity line of the tunica albuginea was pathognomonic for the diagnosis of testicular rupture. Two cases with an inconclusive diagnosis after US were accurately diagnosed as epididymal hematomas. The remaining two cases also showed concordant results with the surgical findings. CONCLUSIONS: MRI can be a useful alternative diagnostic modality for blunt scrotal trauma in limited conditions, especially under circumstances where the use of US infrequently results in an inconclusive diagnosis. PMID- 19131834 TI - Serum procalcitonin levels in patients with multiple injuries including visceral trauma. AB - Procalcitonin (PCT) is known to be a reliable biomarker of sepsis and infection. Elevation of serum or plasma PCT has also been observed after major surgery or trauma. The association of PCT with the severity or location of injury in multiple traumatized (polytrauma) patients has not been clearly established, to date. The aim of this study was therefore to evaluate the sensitivity of PCT as a biomarker for the diagnosis of abdominal trauma. In a prospective clinical study, PCT, interrleukin-6, and C-reactive protein were measured in blood (serum) samples obtained in the emergency room (D0) from 74 patients with multiple injuries and in serum samples obtained on the 2 days after trauma (D1, D2). PCT significantly increased during the first two posttraumatic days in patients with severe multiple injuries (n = 24, day 1: 3.37 ng/mL +/- 0.92 ng/mL; day 2: 3.27 ng/mL +/-0.97 ng/mL) as compared with patients with identical Injury Severity Score but without abdominal injury (day 1: 0.6 ng/mL +/- 0.18 ng/mL; 0.61 ng/mL +/- 0.21 ng/mL). Interrleukin-6 and C-reactive protein serum levels were not able to discriminate between patients with and without abdominal injury during the 2 day posttrauma observation period. In a specific evaluation of the abdominal injury pattern, a significant increase of serum PCT concentrations was observed on day 1 after trauma of the liver (4.04 ng/mL +/- 0.99 ng/mL) and the gut (4.63 ng/mL +/- 1.12 ng/mL) compared with other abdominal lesions (0.62 ng/mL +/- 0.2 ng/mL). Markedly elevated PCT concentrations were also evident after severe multiple injuries, including the liver/spleen in combination with thorax trauma (9.37 ng/mL +/- 2.71 ng/mL). Assessment of serum PCT seems to be significantly increased after abdominal trauma in severe multiple traumatized patients and may serve as a useful biomarker to support other diagnostic methods including ultrasound and CT scan. Although elevated levels of PCT during the first 2 days after trauma are more likely to be indicative of traumatic impact than of an ongoing status of sepsis, multiple events such as surgery, massive transfusion, and intensive care therapy might influence the PCT concentration. PMID- 19131835 TI - Resistin and postburn insulin dysfunction. AB - BACKGROUND: Postburn insulin dysfunction is a significant contributor to morbidity and mortality. A satisfactory mechanism for explaining this phenomenon remains elusive; however, resistin has been postulated to be involved. Initially discovered as an insulin antagonist secreted from adipose tissue in murine models, resistin's function in humans has been more obscure. Resistin is not expressed significantly in human adipocytes although it has been detected in monocytes. We postulate that mononuclear activation at the site of burn injury affects the release of resistin and contributes to insulin dysfunction. METHODS: Plasma from burned and healthy control individuals was characterized for glucose, insulin, and resistin protein levels. Adipose tissue from both groups was analyzed for resistin transcript; levels were found to be somewhat higher in the burned group though not significantly so. Circulating monocyte expression of resistin transcript was assayed in similar fashion. RESULTS: In addition to finding that insulin and glucose were elevated postburn, a finding in agreement with past studies, we demonstrate that circulating resistin levels are significantly elevated as well. Insulin resistance was found to increase at a similar rate to resistin expression in the burn population, suggesting a correlation in these events. Adipose tissue from both groups was analyzed for resistin transcript; levels were found to be somewhat higher in the burned group though not significantly so. Circulating monocyte expression of resistin transcript was assayed and found to be profoundly elevated in the burn population. CONCLUSIONS: This data suggests that resistin is produced by activated monocytes in the adipose tissue around the periphery of burn wound. We suggest that postburn insulin function is adversely affected by resistin produced as a result of this monocyte activation. PMID- 19131836 TI - Inequity of rehabilitation services after traumatic injury. AB - BACKGROUND: The delivery of optimal trauma care involves identification and repair of injuries as well as rehabilitation of patients to return them to their preinjury functional status. This requires access to appropriate rehabilitation services; however, such services may be disproportionately allocated to insured individuals leaving lower income individuals without the means to achieve optimal functional status. We hypothesized that a lack of insurance is associated with a reduced likelihood of being transferred to a rehabilitation facility after trauma. METHODS: A retrospective cohort analysis of blunt National Trauma Databank patients from 2000 to 2004 were assessed to identify predictors of being transferred to a rehabilitation facility at hospital discharge. The likelihood of rehabilitation transfer based on insurance status was assessed, controlling for injury severity, physiology, comorbidities, and patient demographics. RESULTS: Age, gender, comorbidities, injury, presence of blood alcohol at admission, race, and insurance status were independent predictors of being transferred to a rehabilitation facility among blunt trauma patients (n = 750,468). Patients who were uninsured were 53% (OR = 0.47, 95% CI = 0.41-0.53) less likely to be transferred to a rehabilitation facility upon hospital discharge. CONCLUSION: Although access to rehabilitation services after injury is a key predictor of recovery from injury, these services are much less available to uninsured patients. This may lead to significant individual and societal emotional and economic burden. The health and economic impacts of this disparity in access to care need to be investigated to determine the personal and societal impact of appropriate resource allocation to rehabilitation for the uninsured. PMID- 19131837 TI - Adult portal hypertension secondary to posttraumatic extrahepatic portal vein thrombosis treated with Rex shunt. PMID- 19131838 TI - Submammary intercostal arterial injury treated with angioembolization: a case report. PMID- 19131839 TI - Biomechanical and clinical evaluation of posterior malleolar fractures. A systematic review of the literature. AB - INTRODUCTION: Ankle fractures often have involvement of the posterior malleolus. Treatment guidelines exist based on limited biomechanical evidence and still is considered controversial. The objective of this article is to review the biomechanical literature concerning changes in tibiotalar contract area, changes in contact pressures, changes in ankle stability and incongruency of the joint after a posterior malleolar ankle fracture, and to review the clinical literature concerning the outcome of operative and nonoperative treatment of these fractures. Ultimately, the goal is to try to formulate helpful recommendations for clinical practice. MATERIAL: The databases Pubmed/Medline, Cochrane Database of Systematic Reviews, Cochrane Clinical Trial Register, and Embase were searched from 1988 to November 2007 to identify studies relating to changes in tibiotalar contact area, contact pressures, ankle stability, clinical outcome, and radiographic osteoarthritis after a posterior malleolar fracture. The search was restricted to articles written in the English, German, and Dutch language. RESULTS: Eight biomechanical studies, involving 96 cadaveric ankles were included and 10 clinical studies, involving 447 fractured ankles, were included. DISCUSSION: No consensus in the literature was found as to which fragment size of the posterior malleolus (=posterior tibial margin) should be internally fixed. This is partially because of the lack of standardization in examining functional outcomes, making it difficult to compare results. It is not the peak pressure or changes in tibiotalar contact area, but rather the changes in peak pressure distribution that play a large role in posttraumatic arthritis development. It seems important to restore the medial and lateral constraints of the ankle because these, rather than the articular surfaces, provide the majority of ankle stability after an ankle fracture involving the posterior malleolar ankle fracture. PMID- 19131840 TI - Ureteropelvic junction disruption after blunt abdominal trauma. PMID- 19131842 TI - Vulvovaginal atrophy: new and upcoming approaches. PMID- 19131843 TI - Getting to the heart of the matter: asymmetric dimethylarginine and menopause. PMID- 19131844 TI - Disordered folliculogenesis during the menopausal transition: explaining chaos. PMID- 19131845 TI - Vascular effects of estrone and diethylstilbestrol in porcine coronary arteries. AB - OBJECTIVE: To explore the effects of different estrogens on vascular function, we compared the vasorelaxant effects of 17beta-estradiol, 17alpha-estradiol, estrone, and the synthetic estrogen diethylstilbestrol (DES) on porcine coronary arterial segments. DESIGN: Porcine coronary arterial rings were contracted with the stable thromboxane A2 analogue U46619 (3 x 10(-8) M), and direct relaxation was examined by the addition of increasing concentrations of 17beta-estradiol, 17alpha-estradiol, estrone, or DES (10(-9) to 10(-4) M). Modulation of agonist induced contraction and relaxation was studied in coronary arterial rings incubated for 20 minutes with DES or estrone (10(-10)-10(-6) M) with 17beta estradiol (10(-9) M) as comparison. RESULTS: Direct relaxation of arterial rings potentiated by these estrogens was recorded with a rank order potency of DES > 17beta-estradiol > estrone > 17alpha-estradiol. 17beta-Estradiol potentiated relaxation responses to sodium nitroprusside and levcromakalim but not bradykinin or A23187 while reducing contractions to 5-hydroxytryptamine and U46619. DES and estrone, both at 10(-6) M, mimicked the 17beta-estradiol-potentiated sodium nitroprusside and levcromakalim relaxation responses. Additionally, the inhibitory effects of 17beta-estradiol (10(-9) M) on 5-hydroxytryptamine- and U46619-induced contractions were partially reproducible by DES (10(-6) M) and estrone (10(-6) M). CONCLUSIONS: Although DES is the most potent among the tested estrogenic compounds in eliciting relaxation, 17beta-estradiol is more effective than estrone and DES at enhancing endothelium-independent relaxation and reducing vascular contraction in porcine coronary arteries. PMID- 19131847 TI - A U.S. review determined that black cohosh products should be labeled with a cautionary statement. PMID- 19131846 TI - New equol supplement for relieving menopausal symptoms: randomized, placebo controlled trial of Japanese women. AB - OBJECTIVES: Equol, a metabolite of the isoflavone daidzein, is hypothesized to play a major role in the health benefits of soy. We examined the effect of a new S-equol supplement on menopausal symptoms and mood states. DESIGN: We conducted a randomized, double-blind, placebo-controlled trial with our new equol supplement for 12 weeks with 134 Japanese women (aged 40-59 years). They were randomly assigned to three groups: placebo (n = 44), 10 mg of equol per day (EQ-1; n = 44), and 10 mg of equol three times per day (EQ-3; n = 46). Habitual isoflavone intake was limited to 20 mg/d. Participants completed menopausal symptom and Profile of Mood States questionnaires at baseline and postintervention. Physical examination and blood and 24-hour urine collection were performed at baseline and postintervention. RESULTS: At baseline, total menopausal symptom score varied by menopausal and equol producer status (34.3% producers). A total of 127 participants (94.8%) completed the trial. No adverse effects were reported, except for a systemic rash in one EQ-3 woman. The anxiety scores of equol producers were lower than those of nonproducers (P < 0.05). Significant differences between premenopausal and perimenopausal/postmenopausal symptom scores were observed for anxiety, somatic, and total scores. After the EQ-3 intervention, perimenopausal/postmenopausal equol nonproducers showed significant decreases from baseline in all menopausal symptom scores except depression (P < 0.01). Compared with placebo, the EQ-3 group showed significant decreases in depression scores (P < 0.05), as well as significant decreases in Tension-Anxiety (P < 0.05), Depression-Dejection (P < 0.05) and Fatigue (P < 0.01) and increases in Vigor (P < 0.05) of the Profile of Mood States. CONCLUSION: S-equol supplement improved mood-related symptoms in perimenopausal/postmenopausal equol nonproducers. PMID- 19131848 TI - Comment on an article reviewing black cohosh and case reports of hepatotoxicity. PMID- 19131849 TI - Misunderstandings regarding the history and challenges of the Wiley Protocol. PMID- 19131851 TI - Relapse management in multiple sclerosis. AB - Relapses, exacerbations, and attacks are synonymous for new or worsened neurologic symptoms that are the hallmark of relapsing-remitting multiple sclerosis. Management of relapses is not always straightforward. The clinician must distinguish between true relapses, symptom fluctuation, and pseudo-relapses. Risks and benefits of treating a relapse must be considered. Once the decision to treat is made, most clinicians would pursue a course of corticosteroids. Consensus may end there, as there is no clear-cut "best" route of administration or dosing schedule. The patient presenting with their first relapse or clinically isolated syndrome may be at risk for the development of multiple sclerosis. Clinical presentation, CSF findings, and MRI may all give clues as to the risk for future demyelinating events. PMID- 19131852 TI - Molecular imaging studies in Parkinson disease: reducing diagnostic uncertainty. AB - BACKGROUND: The diagnosis of Parkinson disease (PD) is based on clinical criteria but misdiagnosis is as high as 25% of cases as confirmed by anatomic-pathologic studies. Since the introduction of in vivo molecular imaging techniques using Single-Photon Emission Computed Tomography and Positron Emission Tomography, the diagnosis of PD became more reliable by assessing dopaminergic and even nondopaminergic systems. REVIEW SUMMARY: The purpose of this article is to critically review the current data on molecular neuroimaging focusing on the nigrostriatal circuitry and providing useful information on the role of these new imaging techniques in the management of clinically unclear cases of PD. CONCLUSIONS: Patients with essential tremor, psychogenic Parkinsonism or drug induced Parkinsonism can be differentiated from PD in doubtful situations using molecular imaging techniques evaluating striatal dopamine transporters (DAT). However, in patients with vascular Parkinsonism, atypical Parkinsonism and Parkinsonism associated with dementia DAT scans have less diagnostic usefulness. Scans with non-DAT tracers (ie, D2 dopamine receptors) are necessary together with long-term clinical follow-up, and rescans to improve diagnostic accuracy. PMID- 19131853 TI - Atraumatic lumbar puncture needles: after all these years, are we still missing the point? AB - BACKGROUND: Cutting needles remain the most commonly used needle design for lumbar puncture in the neurology community, although atraumatic (noncutting) needles have become common and popular for anesthesiologists performing spinal anesthesia. REVIEW SUMMARY: The use of atraumatic spinal needles for lumbar puncture has been shown to significantly reduce the incidence of postdural puncture headache compared with cutting needles, without loss of efficacy or ease of use. CONCLUSION: The use of noncutting or pencil-point spinal needles should become the standard for performing diagnostic lumbar puncture. PMID- 19131854 TI - Paraneoplastic neurological syndromes: a review. AB - BACKGROUND: Paraneoplastic neurologic syndromes (PNS) constitute a rare group of disorders resulting from damage to the nervous system in the setting of cancer physically unrelated to the tumor site. PNS are believed to result from an autoimmune attack of normal neuronal tissue, spurred by similar neuronal antigens ectopically expressed by tumor cells. REVIEW SUMMARY: The most common PNS are reviewed and also their association with specific onconeural antibodies, some directly pathogenic, others whose role in the disease process is less clear-cut. This diversity in pathogenesis is likely due to the relative role of humoral versus cellular immunity in PNS. Virtually any cancer may result in PNS but certain tumors, small cell lung cancer, gynecologic cancers (breast and ovarian), thymoma, and plasma cell tumors are more frequently encountered. In most instances, immunosuppressive therapy is unhelpful and outcome is poor. CONCLUSIONS: PNS have diverse presentations, affecting both the central and peripheral nervous system and commonly, it is the PNS, not cancer that is the presenting symptom. Only subsequently, after onconeural antibodies are discovered or cancer is found, is PNS diagnosed. Neurologists should familiarize themselves with these rare syndromes and treatment principles, as rapid detection and treatment of the underlying tumor offer the best chance for recovery or prevention of further neurologic deterioration. PMID- 19131855 TI - Othello syndrome in Parkinson disease patients without dementia. AB - BACKGROUND: Delusional jealousy or Othello syndrome (OS) is a well-described psychiatric disorder with paranoid features reported in both organic and functional psychoses. In organic psychoses, the disorder occurs more frequently among chronic male alcoholics and in demented patients. To date, only 2 anecdotal cases of OS have been reported in Parkinson disease (PD) during dopaminergic treatment. OBJECTIVE: To investigate the presence of OS in PD patients and to study the relationship between dopaminergic treatment, avoiding the possible influence of dementia. METHODS: Five hundred sixty-three PD patients without dementia encountered in our movement disorders practice were included in the study. All patients who developed OS were studied. Relationships between clinical and familial history and dopaminergic therapy and OS were assessed. RESULTS: Six patients with OS were identified. They were all male, with a relatively recent diagnosis of PD characterized by mild-moderate motor deficit. Dopaminergic treatment had been prescribed at low dosages. Neither confusional states (including agitated confusion) nor delirium were associated with OS. The disorder became manifest mainly at time of introduction/increment of antiparkinson treatment. Invariably, OS decreased or receded after reduction/suspension of the antiparkinson drug and prescription of an atypical neuroleptic, usually clozapine or quetiapine. CONCLUSION: We hypothesize that nondemented PD patients affected by OS do not necessarily present with severe motor complications and may well have a biologic predisposition for psychiatric disorders. In our opinion this paranoid delusion is rarely considered in PD. PMID- 19131856 TI - Leopard syndrome and Chiari type I malformation: a case report and review of the literature. AB - INTRODUCTION: The authors present a 31-year-old man with a Chiari type I malformation (CM-1) occurring in conjunction with LEOPARD syndrome. He presented with severe dysphagia requiring placement of a percutaneous endoscopic gastrostomy (PEG) tube. Evaluation included magnetic resonance imaging (MRI) of the brain and cervical spine that revealed CM-1 with an extensive cervical syrinx. The patient underwent a suboccipital craniectomy with C1 laminectomy and duraplasty. His symptoms quickly resolved and his PEG tube was removed. DISCUSSION: The occurrence of a CM-1 with LEOPARD syndrome has only been reported once, whereas CM-1 and Noonan syndrome have been linked in several cases. The similarity between LEOPARD and Noonan syndromes has been reported and many propose they represent 2 entities along a spectrum. CONCLUSION: In light of this spectrum, we propose that CM-1 should be considered in all patients presenting with LEOPARD-Noonan syndrome. PMID- 19131857 TI - Nerve rootlet and fascicular biopsy in patients with hypertrophic inflammatory neuropathy. AB - BACKGROUND: The differential diagnosis of hypertrophic mononeuropathy includes multiple diagnoses with significantly differing treatments and prognosis. The value of pathologic confirmation of the disease entity is critical in defining optimal patient care. REVIEW SUMMARY: This report is an analysis of 2 unusual cases of progressive neuropathy with radiographic evidence of peripheral nerve enlargement. The clinical presentation of the patients and diagnostic value of the nerve biopsy are discussed. The aim of this report is to alert and emphasize the importance of a peripheral nerve biopsy in patients with hypertrophic mononeuropathies and carefully selected patients with hypertrophic polyradiculopathies. Both patients were treated with immunomodulatory agents with modest improvement in their symptoms. CONCLUSION: Pathologic confirmation of chronic inflammatory demyelinating polyradiculopathy in these patients led to the correct diagnosis and appropriate treatment. Surgical excisional biopsy of a lumbar nerve rootlet and fascicular biopsy of the sciatic nerve were necessary to secure proper diagnosis in these cases. Neither patient had any complications or sequelae from the procedures. PMID- 19131858 TI - Stroke due to bee sting. AB - Although bee stings can cause local reactions, neurovascular complications are rare. A 60-year-old man developed a focal neurologic deficit 2 hours after multiple bee stings, which was confirmed to be acute cerebral infarctions on magnetic resonance imaging scan. PMID- 19131859 TI - Serum neuron specific enolase to predict neurological outcome after cardiopulmonary resuscitation: a critically appraised topic. AB - BACKGROUND: Despite a large body of resuscitation research, the optimal timing and approach to prognosticating futility after nontraumatic cardiac arrest remains controversial. Postresuscitation anoxic-ischemic encephalopathy may leave patients cognitively disabled and dependent, minimally conscious, or in a persistent vegetative state. Neurologists are frequently called upon to assess comatose postcardiac arrest patients to communicate accurate prognostic information and to assist in planning the most appropriate level of care. Serum neuron specific enolase (NSE), when used in conjunction with other available prognostic tools, may be a useful tool for predicting clinical outcome in this patient population. OBJECTIVE: Determine the clinical utility of a serum NSE measurement for predicting the neurologic outcome of a survivor after resuscitation from a nontraumatic cardiac arrest. METHODS: We addressed the question through the development of a structured, critically appraised topic. Participants included consultant and resident neurologists, clinical epidemiologists, medical librarian, and clinical content experts in the fields of emergency medicine, cardiac resuscitation, and critical care neurology. Participants started with a clinical scenario and a structured question, devised search strategies, located and compiled the best evidence, performed a critical appraisal, synthesized the results, summarized the evidence, provided commentary, and declared bottom-line conclusions. RESULTS: A single recent prospective cohort study, which assessed the predictive value of serum NSE was selected and appraised. A level of 80 ng/mL was determined to predict persistent coma with a sensitivity of 63% (95% CI, 49%-75%), and a specificity of 100% (95% CI, 97% 100%), positive predictive value (PPV) of 100%, negative predictive value (NPV) of 84%, and a negative LR of 0.37. CONCLUSIONS: The specificity of serum NSE levels >80 ng/mL is sufficiently high that, when it is used with other clinical and electrophysiological data, it could be useful as a prognostic indicator of neurologic outcome after cardiac arrest. Although serum NSE is an appealing, simple, readily available test, prediction of neurologic outcome after resuscitation from cardiac arrest must not rely solely on a serum biomarker, but must be determined in the context of other patient characteristics and neurologic examination findings. PMID- 19131860 TI - Motor neuron disease. PMID- 19131861 TI - An unusual cause of cerebral cardioembolism in a 33-year-old man due to ventricular noncompaction. PMID- 19131862 TI - An unusual cause of cardioembolic stroke: isolated left ventricular noncompaction. PMID- 19131863 TI - Proximal splenic artery embolization in the management of splenic rupture. AB - OBJECTIVE: To report the use of proximal splenic artery embolization for management of spontaneous splenic rupture. DESIGN: Case report and literature review. SETTING: A tertiary pediatric critical care unit in a university teaching hospital. INTERVENTIONS: Proximal splenic artery embolization. MEASUREMENTS AND MAIN RESULTS: An 8-yr-old boy presented with abdominal pain radiating to the left shoulder 9 days after completing induction chemotherapy for acute lymphoblastic leukemia. Imaging revealed a splenic rupture with parenchymal and subcapsular hematomas, with no evidence of active extravasations. The patient was admitted to the pediatric critical care unit for close hemodynamic monitoring and frequent measurements of hemoglobin. His lowest recorded hemoglobin and hematocrit were 63 g/L and 0.19 L/L, respectively. Posttransfusion of packed red blood cells, he was taken to interventional radiology for proximal splenic artery embolization under moderate sedation. Several coils were successfully placed in the proximal splenic arterial system resulting in a marked reduction of splenic blood flow without disruption of collaterals. The patient recovered well from proximal splenic artery embolization in the pediatric critical care unit and experienced short lasting abdominal pain and fever for 1 day. He was discharged home 4 days after the procedure and follow-up imaging showed resolving hematomas with preserved splenic blood flow. CONCLUSION: Proximal splenic artery embolization in children may be a safe therapeutic alternative to either conservative or surgical management in spontaneous splenic rupture. Preservation of splenic tissue with a reduced risk of repeated hemorrhage can be obtained with proximal splenic artery embolization. PMID- 19131864 TI - Septic encephalopathy complicating acute appendicitis. PMID- 19131865 TI - Near infrared spectroscopy changes with pericardial tamponade. AB - OBJECTIVE: Report on the use of near infrared spectroscopy (NIRS) for the recognition of pericardial tamponade after neonatal congenital heart surgery. DESIGN: Case report. SETTING: Pediatric cardiac intensive care unit. PATIENT: Seven-day-old term infant with double inlet left ventricle, hypoplastic aortic arch, and ascending aorta, postoperative day (POD) 3 from the Norwood procedure. INTERVENTION: After an initial stable early postoperative course, the patient's cerebral saturation decreased from the 50s on POD 2, to the 40s in the early morning hours of POD 3. By 9 am on POD 3, the cerebral saturation decreased further to the mid 30s. No change in pulsoximetry, arterial blood gas acid base balance, or blood pressure occurred. An echocardiogram was performed due to the progressive decline in cerebral saturation values. A 10-mm circumferential pericardial effusion was diagnosed. The effusion was drained without incident. Cerebral saturation returned to the 50s, the patient had no further complications and was discharged to home on POD 15. CONCLUSION: Pericardial tamponade is a well known complication of open heart surgery in children, and early recognition of this can be difficult. This case report demonstrated cerebral saturation to be decreased as impending pericardial tamponade developed. Along with the commonly used markers of tamponade, near infrared spectroscopy measurement of cerebral saturation may also be of benefit in recognizing this life-threatening condition. PMID- 19131866 TI - Reversible acute left ventricular dysfunction in accidental strangulation. AB - OBJECTIVE: : To report reversible acute left ventricular dysfunction after accidental strangulation in an 8-year-old boy. DESIGN: : Case report. SETTING: : Pediatric intensive care unit in a tertiary care teaching hospital. PATIENT: : Review of the medical record and clinical course of the patient who developed cardiogenic shock and pulmonary edema after strangulation. INTERVENTIONS: : Mechanical ventilation, monitoring of cardiovascular status, and serial evaluation of cardiac enzymes. MEASUREMENTS AND MAIN RESULTS: : In the acute period, electrocardiogram showed transient global ST elevation with rise of cardiac enzymes and global left ventricular hypokinesia. Hemodynamic status improved over 24 hrs with resolution of electrocardiogram findings and left ventricular dysfunction. CONCLUSIONS: : Exaggerated sympathetic stimulation due to strangulation can result in acute reversible myocardial dysfunction mimicking myocardial infarction. PMID- 19131867 TI - Right pneumonectomy syndrome presenting as a cardiac arrest in a two-year-old. AB - INTRODUCTION: Pneumonectomy syndrome is a rare complication after pneumonectomy with variable onset and presentation. It is characterized by extrinsic airway compression secondary to mediastinal shift. As far as we are aware, we are reporting the first pediatric case of pneumonectomy syndrome presenting as a cardio-pulmonary arrest. PATIENT AND INTERVENTION: A two-year-old boy presented with a cardiac arrest 8 months after his pneumonectomy. He failed extubation twice. A flexible bronchoscopy was then performed which revealed external compression of the left mainstem bronchus. He underwent surgical correction with a prosthetic, thoracic implant. RESULT: The patient's trachea was subsequently extubated to room air on postoperative day 1. He remains clinically stable at 6 month follow-up. CONCLUSION: Postpneumonectomy syndrome is a rare but serious complication after pneumonectomy. If the airway compression occurs in conjunction with compromised vena caval return, cardio-pulmonary arrest can occur. Therefore, timely diagnosis and early intervention is required. PMID- 19131869 TI - Do we know when to say "wean"? PMID- 19131870 TI - Clinical decision support systems: Important tools when appropriately used. PMID- 19131871 TI - Zinc-Jack of all trades, master of none! PMID- 19131872 TI - Dysharmony of hormonal functions in critically ill. PMID- 19131873 TI - Outcome assessment--what outcome do we want? PMID- 19131875 TI - Broad-spectrum antibiotic use in the pediatric intensive care unit: Balancing patient interests against intensive care unit ecology. PMID- 19131876 TI - ECHO for ECMO: not just for cardiac function. PMID- 19131877 TI - Pulmonary function and shunt size after single ventricle palliation--classic cardiopulmonary interactions. PMID- 19131878 TI - Resembling but not equal. PMID- 19131879 TI - Metabolic acidosis, respiratory distress, and children with severe acute asthma. PMID- 19131881 TI - Support for PICUs and the PRISM scoring algorithms. PMID- 19131882 TI - More on perfusion index and its use as a maternal monitor for fetal well being. PMID- 19131884 TI - Determinants of nonadherence to a single-dose nevirapine regimen for the prevention of mother-to-child HIV transmission in Rwanda. AB - OBJECTIVES: To describe experiences, and identify factors associated with nonadherence to a single-dose nevirapine (SD-NVP) regimen for the prevention of mother-to-child transmission (PMTCT) of HIV in Rwanda. METHODS: In April to May 2006, using a case-control design at 12 PMTCT sites, we interviewed HIV-infected women who did not adhere (n = 111) and who adhered (n = 125) to the PMTCT prophylaxis regimen. Nonadherence was defined as mother and/or infant not ingesting SD-NVP at the recommended time or not at all and adherence as mother infant pairs who ingested it as recommended. RESULTS: Only 61% of nonadherent women had received SD-NVP during pregnancy or delivery. Among nonadherent women who received SD-NVP, 80% ingested it at the recommended time, representing 49% of all nonadherent women. Only 7% of their newborns ingested SD-NVP. Multivariate logistic regression showed that unmarried women, less educated women, women who made 2 or less antenatal care visits, and those offered HIV testing after their first antenatal care visit were more likely to be nonadherent to PMTCT prophylaxis. Not disclosing one's HIV status to someone aside from a partner was also associated with nonadherence in mother-infant pairs. CONCLUSIONS: Sociodemographic factors, health services delivery factors, and a lack of communication and social support contributed to nonadherence to PMTCT prophylaxis in Rwanda. PMID- 19131885 TI - Test and tell: correlates and consequences of testing and disclosure of HIV status in South Africa (HPTN 043 Project Accept). AB - BACKGROUND: As the numbers of HIV-positive diagnoses rise in South Africa, it is important to understand the determinants and consequences of HIV disclosure. METHODS: Cross-sectional survey from random community samples of men and women in urban and rural South Africa (n = 217 HIV-positive individuals, 89% female). RESULTS: Two thirds of all known HIV-infected adults in these communities had disclosed their status to sexual partner(s). On average, individuals who disclosed were 2 years older, higher in socioeconomic assets, and had known their HIV status 7 months longer than those who had not told their sexual partner(s). The "need for privacy" was the most cited reason (45%) for nondisclosure among those who had never disclosed. People who eventually disclosed their HIV status to sexual partner(s) were significantly more likely to report always or more frequently using condoms, reducing their number of sexual partners, and/or becoming monogamous. Among individuals who disclosed their HIV status, 77% reported increases in social support, with families providing the most support. CONCLUSIONS: Disclosure is associated with reports of consequent safer sexual behavior and greater social support. Interventions might be informed by the costs and benefits of disclosure and differences in disclosure to sexual partner vs. to one's social network. PMID- 19131887 TI - Impact of HIV-1 infection on the hematological recovery after clinical malaria. AB - BACKGROUND: Anemia is the most frequent cytopenia in HIV-infected individuals and is often associated with malaria. OBJECTIVE: To assess the impact of HIV-1 on the hematological recovery after a clinical malaria episode. METHODS: In Ndola, Zambia, a region with high malaria and HIV prevalence, hemoglobin (Hb) was measured in 634 malaria patients 14 and 45 days after antimalarial treatment. Risk factors for hematological recovery were analyzed in a multivariate linear regression model. RESULTS: At enrollment, HIV-1-infected malaria patients had lower Hb compared with HIV-1 uninfected (122.7 vs 136.0 g/L; P < 0.001). In both groups, mean Hb was significantly lower at day 14 posttreatment than day 0 (P < 0.0001) and significantly higher at day 45 than at day 14 (HIV-1 negative: P = 0.0001; HIV-1 infected: P = 0.005). HIV-1 was a risk factor for a larger Hb decrease until day 14 (P < 0.001) and slower recovery until day 45 (P = 0.048). When considering the whole 45-day follow-up period, mean Hb increased in the HIV 1-negative group (+3.54 g/L; 95% confidence interval: 1.37 to 5.70; P = 0.001) but not in the HIV-1-infected group (-0.72 g/L; 95% confidence interval: -3.85 to +2.40; P = 0.64). HIV-1 infection as such (P < 0.0001), not CD4 cell count (P = 0.46), was an independent risk factor for a slower hematological recovery. CONCLUSIONS: HIV-1-infected malaria patients had a slower hematological recovery after successful parasite clearance. Malaria preventive measures should be targeted to this high-risk group. PMID- 19131888 TI - Intensified case finding for tuberculosis in prevention of mother-to-child transmission programs: a simple and potentially vital addition for maternal and child health. PMID- 19131889 TI - Changes in darunavir/r resistance score after previous failure to tipranavir/r in HIV-1-infected multidrug-resistant patients. AB - OBJECTIVES: To evaluate changes in resistance to tipranavir/r (TPV/r) and darunavir/r (DRV/r) in patients who had failed a TPV/r-including regimen. METHODS: HIV genotypes obtained both at baseline and at tipranavir/r failure (TPVF) were analyzed in 47 HIV-infected patients failing a TPV/r-including regimen. Genotypes were evaluated through the Stanford mutation score: patients were ranked for TPV/r and DRV/r resistance as susceptible (class 1), potential low-level (class 2), low-level (class 3), intermediate-level (class 4), and high level resistance (class 5). Values are expressed as median (interquartile range) or as frequency (%). RESULTS: Forty-seven patients were eligible. At baseline (tipranavir initiation), the scoring for TPV/r was: class 3 = 4 (8.5%); class 4 = 31 (66%); and class 5 = 12 (25.5%). Corresponding scores for DRV/r were: class 2 = 1 (2%), class 3 = 12 (25.5%), class 4 = 32 (68%), and class 5 = 2 (4.5%). At TPVF, a shift toward a higher TPV/r scoring class was seen in 16 (34.1%) patients (P = 0.001), whereas a shift toward a higher DRV/r scoring class was observed in 9 (19.2%) patients (P = 0.2381). After TPVF, 25/47 patients (53%) were treated with a DRV/r-containing regimen. After 24 weeks (on-treatment analysis), the median HIV RNA decrease was 3.04 (2.13-3.45) log10 copies per milliliter in DRV/r group versus -0.04 (-0.44; 0.50) log10 copies per milliliter in patients not treated with a DRV/r-containing regimen (P < 0.0001); CD4 increase was 126 (70 169) cells per cubic millimeter in DRV/r group versus -42 (-121; 42) (P < 0.0001). DISCUSSION: Treatment with TPV/r did not significantly increase the resistance score to DRV/r and did not preclude the efficacy of subsequent treatment with DRV/r. PMID- 19131886 TI - Interleukine-2 therapy does not increase the risk of Hodgkin or non-Hodgkin lymphoma in HIV-infected patients: results from FHDH ANRS CO4. AB - BACKGROUND: Concerns have been raised about a possible excess risk of lymphomas in HIV-infected patients exposed to interleukin 2 (IL-2) therapy. Here we compared the risks of non-Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL) in IL 2-treated and IL-2-untreated HIV-infected patients. METHODS: Patients monitored through the French Hospital Database on HIV between May 1, 1995, and December 31, 2005, were enrolled in this study. Lymphomas that occurred between the day after study entry and the end of follow-up were eligible for analysis. Poisson regression models were used in 2 separate analyses to quantify the possible relationship between IL-2 therapy and the incidence of NHL and HL. RESULTS: The IL-2-treated group consisted of 861 patients and the IL-2-untreated group of 77,605 patients. Follow-up lasted a total of 3643 and 382,720 person-years, respectively. After adjustment for sex and time-updated age, period, the CD4 cell counts, the plasma HIV RNA levels, and AIDS status, the relative rates of NHL and HL associated with IL-2 therapy were 0.64 (95% confidence interval, 0.25 to 1.65) and 0.33 (95% confidence interval, 0.04 to 2.86), respectively. CONCLUSIONS: In this large observational study, IL-2 therapy did not increase the risk of lymphoma, either NHL or HL, in HIV-infected patients. PMID- 19131890 TI - Short-term and long-term effects of highly active antiretroviral therapy on the incidence of herpes zoster in HIV-infected children. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) has reduced herpes zoster (HZ) incidence in HIV-infected children, yet it remains common. METHODS: We evaluated perinatally HIV-infected youth with varicella infection enrolled between 1993 and 2006 in a prospective cohort study. Incidence rates (IRs) and 95% confidence intervals of HZ were estimated by calendar year, age group, and HAART use. The effect of initiating HAART was also evaluated by fitting Cox survival models adjusted for potential confounders. RESULTS: Among 536 perinatally infected children with documented prior varicella (median follow-up = 6.8 years), 116 (22%) developed HZ (IR = 3.2 events/100 person-years, confidence interval: 2.6 to 3.8). IRs increased from 1993 to 1996 and then declined significantly through 2006 (P < 0.001). However, an IR of 1.4-3.1 HZ episodes per 100 person-years persisted from 2001 to 2006. The risk of HZ was higher for those with lower CD4% or in Centers for Disease Control and Prevention clinical class C. The IR of HZ was similar in the 90 days before or after initiation of HAART but declined significantly after more than 90 days of HAART. CONCLUSIONS: Although HAART has markedly reduced the IR of HZ, it remains a frequent complication in HIV-infected children. The risk of HZ is similar in the 90 days before and after initiating HAART. PMID- 19131891 TI - Persistence of virological benefits following directly administered antiretroviral therapy among drug users: results from a randomized controlled trial. AB - BACKGROUND: Although directly administered antiretroviral therapy (DAART) has demonstrated impressive biological benefits compared with self-administered therapy (SAT) among drug users, the persistence of DAART after transition to SAT has not been examined. METHODS: We conducted a community-based, prospective, randomized controlled trial of 6 months of DAART compared with SAT. The primary outcome was the proportion of subjects who achieved virological success at 6 months postintervention, defined as either a 1.0 log10 reduction from baseline or HIV-1 RNA <400 copies per milliliter. Secondary outcomes included the change from baseline in HIV-1 RNA and CD4 lymphocyte count. RESULTS: Of the 53 subjects in the SAT arm and 88 subjects in the DAART arm, 52 and 82, respectively, provided blood samples at 6 months postintervention. The DAART (n = 88) and SAT (n = 53) arms did not differ on virological success (DAART 58.0% vs. SAT 56.6%, P = 0.64), mean reduction in log10 HIV-1 RNA (-0.79 vs. -0.31 log10 copies/mL, P = 0.53), or mean change in CD4 lymphocyte count (+60.2 vs. -15.4 cells/mL, P = 0.12). In the multivariate analysis, only high levels of social support significantly predicted virological success. CONCLUSIONS: This analysis, from the first randomized controlled trial of DAART among active drug users, failed to show the persistence of the DAART intervention at improving virological outcomes. Additional strategies are needed to ensure the on-treatment benefits persist following the cessation of DAART. PMID- 19131892 TI - Epidemiological data of different human papillomavirus genotypes in cervical specimens of HIV-1-infected women without history of cervical pathology. AB - AIM: To study the epidemiology of different human papillomavirus (HPV) genotypes in cervical samples of HIV-1-infected women with normal Papanicolau smears. DESIGN: : Retrospective analysis of a prospective cohort. PATIENTS AND METHODS: We selected HIV-1-infected women with 2 consecutive normal Papanicolau smears at baseline and at least 1 baseline and 1 follow-up cervical sample. HPV infection was assessed by second-generation hybrid capture (HC-2) and multiplex polymerase chain reaction (mPCR). HPV genotypes were determined by mPCR. RESULTS: From a cohort of 139 women followed up to 4 years, 93 women meeting the inclusion criteria were analyzed. The mean period between samples was 20 months (range, 6 44 months). HPV baseline prevalence was 63% [59/93; 95% confidence interval (CI), 53% to 73%] using polymerase chain reaction and 41% (38/93; 95% CI, 31% to 51%) using HC-2, P = 0.007 (kappa, 0.45; P = 0.001). The most prevalent high oncogenic risk genotypes (HR-HPV) were HPV-16 (28%), HPV-33 (18%), HPV-52 (12%), HPV-58 (11%), and HPV-39 (11%). Infection with multiple HPV genotypes was detected in >40% of women. HPV infection persisted at follow-up in 86% (51/59; 95% CI, 77% to 95%) by polymerase chain reaction and 76% (29/38; 95% CI, 62% to 90%) by HC-2. HPV infection persisted in 55% of women with samples available beyond 3 years. The actuarial probabilities of clearance and incidence of HPV infection at 36 months were 16% and 45%, respectively. CONCLUSIONS: HPV infection is highly prevalent and persistent among HIV-1-infected women with normal Papanicolau smears. HR-HPV genotypes other than HPV-16 (HPV-33, HPV-52) are frequently detected in HIV-infected women. mPCR provides better surveillance of HPV infection than HC-2 methods. PMID- 19131894 TI - Carotid intima-media thickness and arterial stiffness in HIV-infected patients: the role of HIV, antiretroviral therapy, and lipodystrophy. AB - OBJECTIVES: HIV-infected patients using combination antiretroviral therapy (ART) have an increased cardiovascular risk. We aimed to identify the effects of HIV, ART, and lipodystrophy (LD) on carotid artery intima-media thickness (C-IMT), a surrogate measure of atherosclerosis, and arterial stiffness, a marker of cardiovascular risk. DESIGN: Case-control study of 77 HIV-infected men (55 exposed to ART, 22 ART naive, and 23 with LD) and 52 controls. METHODS: C-IMT was measured ultrasonically, and arterial stiffness was estimated by distensibility (DC) and compliance (CC) coefficients of the carotid, femoral, and brachial arteries, by the carotid Young elastic modulus and pulse wave velocity. RESULTS: After adjustment for cardiovascular risk factors, HIV-infected patients had a 0.067 mm (10.8%) greater C-IMT than controls, 13.6% and 29.5% lower DC, and 14.1% and 31% lower CC of the carotid and femoral arteries, respectively, but similar Young elastic modulus and pulse wave velocity. Patients exposed to ART had similar C-IMT compared with ART-naive patients but 25.9% lower DC and 21.7% lower CC of the femoral artery. Arterial properties did not differ between patients with and without LD. CONCLUSIONS: HIV infection is independently associated with C-IMT and generally increased arterial stiffness. ART use is associated with increased stiffness of the femoral artery. PMID- 19131893 TI - Use of saliva as a lubricant in anal sexual practices among homosexual men. AB - OBJECTIVES: Compared with other sexually active adults, men who have sex with men (MSM) are more frequently infected with several pathogens including cytomegalovirus, hepatitis B virus, and Kaposi sarcoma-associated herpesvirus. Because one common element between these organisms is their presence in saliva, we evaluated saliva exposure among MSM in a heretofore relatively unrecognized route-via use of saliva as a lubricant in anal sex. METHODS: MSM in a San Francisco population-based cohort were interviewed regarding use of saliva by the insertive partner as a lubricant in various anal sexual practices. RESULTS: Among 283 MSM, 87% used saliva as a lubricant in insertive or receptive penile-anal intercourse or fingering/fisting at some point during their lifetime; 31%-47% did so, depending upon the act, in the prior 6 months. Saliva use as a lubricant was more common among younger men and among HIV-infected men when with HIV-infected partners. Even among MSM following safe sex guidelines by avoiding unprotected penile-anal intercourse, 26% had anal exposure to saliva via use as a lubricant. CONCLUSIONS: Among MSM, use of saliva as a lubricant is a common, but not ubiquitous, practice in anal sex. The findings provide the rationale for formal investigation of whether saliva use in this way contributes to transmission of saliva-borne pathogens in MSM. PMID- 19131895 TI - Effect of simultaneous use of highly active antiretroviral therapy on survival of HIV patients with tuberculosis. AB - INTRODUCTION: The optimal timing for initiation of highly active antiretroviral therapy (HAART) in patients with AIDS and tuberculosis (TB) is an unresolved question. To assess the effect of HAART on the survival of patients with TB, we designed this study. METHODS: We selected all HIV patients included in the COMESEM cohort with TB diagnosis after 1996. Clinical and epidemiological data were registered. We compared patients who started HAART at the diagnosis of TB [simultaneous therapy (ST)] or not. Survival was assessed by Cox analysis. RESULTS: Among the 6934 HIV patients included in the cohort, 1217 patients had TB, 322 of them (26.5%) after 1996. At the time of TB diagnosis, 45% of them started HAART (ST). There were no differences between groups regarding basal characteristics, except for a lower viral load in ST patients. ST therapy was associated with improved survival (hazard ratio 0.38; 95% confidence interval 0.20 to 0.72, P = 0.003). By univariate analysis, survival was also associated with no endovenous drug use and a later year of TB diagnosis. After adjusting for other prognostic variables, by Cox multivariate analysis, ST remained robustly associated with improved survival (hazard ratio 0.37; 95% confidence interval 0.17 to 0.66, P = 0.001). CONCLUSIONS: Simultaneous HAART and TB treatment in HIV patients with TB is associated with improved survival. PMID- 19131896 TI - Premature aging of T cells is associated with faster HIV-1 disease progression. AB - OBJECTIVE: To determine if untreated HIV-1 infection and progression is associated with premature aging of memory CD8 and CD4 T cells and naive CD4 T cells. METHODS: Twenty HIV-1-infected fast progressors and 40 slow progressors were included in our study, using risk set sampling. The expression of cell surface markers reflecting the differentiation stages of lymphocytes was measured using flow cytometry analyses performed on cryopreserved peripheral blood mononuclear cells. RESULTS: We found that HIV-1 disease progression is associated with a decreased CD28 median florescence intensity on CD4 and CD8 T cells; an increased proportion of intermediate- and late-differentiated CD8 T cells and a decreased CD31 median florescence intensity on naive CD4 T cells of recent thymic origin. A selective depletion of peripherally expanded naive CD4 T cells was found to be associated with HIV-1 infection but not with HIV-1 disease progression. CONCLUSIONS: The overall change during HIV-1 infection and progression is associated with a shift in the T-cell population toward an aged conformation, which may be further compromised by impaired renewal of the less differentiated CD4 T-cell population. Our results suggest that HIV-1 infection induces an accelerated aging of T lymphocytes, which is associated with the clinical progression to AIDS and death. PMID- 19131898 TI - Estimation of the predictive role of plasma viral load on CD4 decline in HIV-1 subtype C-infected subjects in India. AB - BACKGROUND: Plasma viral load has been shown to be a meaningful prognostic marker for disease progression in untreated, HIV-1 subtype B-infected subjects in United States and Western Europe and therefore used as a prognostic marker for disease progression. Because of high expenses of commercially available viral load assays, the role of viral load in disease progression has not been evaluated in HIV-1 subtype C-infected patients in India. METHODS: We developed an inexpensive real-time reverse transcriptase-polymerase chain reaction assay to quantify viral load in plasma of HIV-1 subtype C-infected subjects from India and used it in a longitudinal analysis of viral load and CD4 cell number in HIV-infected subjects from Calcutta, India. RESULTS: The real-time reverse transcriptase-polymerase chain reaction assay can quantify plasma viral load with a linear range of detection from 10 to 10 HIV-1 RNA copies per input. Longitudinal analysis of viral load in a cohort of 39 subjects over an average period of approximately 3 years indicates that 1-log increase in HIV-1 RNA level was associated with a decline of 67 CD4 cell count. Furthermore, HIV-1 RNA level between 500 and 50,000 copies per milliliter would predict a 12.9% decrease in CD4 cell count per year, whereas HIV-1 RNA levels above 50,000 copies HIV-1 RNA per milliliter would predict a 25.3% decrease in CD4 cells per year. In addition, we estimated that the mean incubation period of disease development, as defined by the loss of CD4 below 200, is 8.2 years. CONCLUSION: Our report on the level of viral load on predicting CD4 decline in Indian subjects with HIV-1 provides an additional important tool to the physicians for treating and planning a therapeutic strategy to control HIV-1 infection in India. PMID- 19131899 TI - Auditory nerve neuropathy in a neonate after linezolid treatment. PMID- 19131900 TI - Breastfeeding prevents severe disease in full term female infants with acute respiratory infection. AB - BACKGROUND: Breastfeeding is a well-known protective factor against severe respiratory tract infections. Recently, a gender specific role for human milk has been described in very low birth weight infants and neonates: breast milk protected girls but not boys. OBJECTIVE: To determine whether the protective effect of breastfeeding on the severity of acute respiratory infections in full term infants is different for girls and boys. METHODS: A prospective cross sectional study of infants seeking medical care for acute respiratory infection. The protective role of breastfeeding against viral pneumonia and hospitalization were assessed by univariate and multivariate analyses. Analyses were adjusted for important confounders. RESULTS: A total of 323 patients were enrolled in this study. Breastfeeding protected girls against pneumonia and hospitalization, but did not protect boys. Nonbreastfeeding females were particularly susceptible to severe acute respiratory infections. CONCLUSIONS: Breastfeeding had a protective effect against severe disease in infant girls experiencing their first symptomatic respiratory infection. Nonbreastfeeding females are at significant risk for severe acute lung disease and should be targeted intensively by breastfeeding campaigns. PMID- 19131901 TI - Trends in otitis media and myringtomy with tube placement among American Indian/Alaska native children and the US general population of children. AB - BACKGROUND: Otitis media (OM) morbidity in American Indian and Alaska Native (AI/AN) children is historically higher than that in other US children. METHODS: Outpatient visits and hospitalizations listing OM as a diagnosis and outpatient visits listing myringotomy with insertion of tubes as a procedure among AI/AN children <5 years of age from the Indian Health Service National Patient Information Reporting system for 2003-2005 were analyzed. Outpatient visits and hospitalizations with OM for the general US child population were analyzed using the National Hospital Ambulatory Medical Care and National Ambulatory Medical Care Surveys for 2003-2005, and the 2003 Kids' Inpatient Database, respectively. RESULTS: The OM-associated outpatient visit rate for AI/AN children <5 years of age (89 per 100 children/yr) for 2003-2005 was less than that reported for 1994 1996 (138); however, the rate increased for Alaska region (158 to 181). The OM outpatient visit and myringotomy with insertion of tubes rates (181 and 2.6 per 100 children/yr, respectively) for AI/AN children in Alaska were higher than rates for children in each of the other IHS regions and rates for US children (63 and 1.8 per 100 children/yr, respectively). The OM outpatient visit rates for AI/AN infants (184), especially in the Alaska region (334), were higher than the rate for US infants (84). CONCLUSIONS: The OM-associated outpatient visit rate in AI/AN children <5 years of age has decreased but remains higher than that of the US general child population; however, the rate increased in the Alaska region, where a limited decline in invasive pneumococcal disease has been demonstrated. The ongoing disparity in OM outpatient visit rates among AI/AN children, especially Alaska Native children, indicates a need for new prevention measures, including expanded-valency pneumococcal conjugate vaccines, to reduce OM morbidity. PMID- 19131897 TI - CXCR4-using HIV type 1 variants are more commonly found in peripheral blood mononuclear cell DNA than in plasma RNA. AB - OBJECTIVE: To compare the distribution of R5-like and X4-like HIV-1 envelope sequences in plasma and peripheral blood mononuclear cell (PBMC). METHODS: Clonal sequencing of the HIV-1 glycoprotein 120 region was performed on PBMC DNA and plasma RNA of 11 HIV-1 subtype B-infected patients with high probability of carrying X4 virus. Coreceptor use was predicted using the position-specific scoring matrix (PSSM). RESULTS: A total of 330 and 427 clonal envelope sequences were obtained from PBMC and plasma, respectively. PSSM interpretation revealed the presence of a mixture of predicted X4 and R5 sequences in 10 patients and pure R5 sequences in 1. The X4 sequences were significantly more represented in PBMC (with an average of 52.2% of the clonal proviral sequences scored X4) compared with plasma (19.7% X4 sequences) (P < 0.0001). At the single patient level, the higher representation of X4 sequences in PBMC reached statistical significance (P < 0.002) in 6 individuals. CONCLUSIONS: Mixtures of X4 and R5 sequences with highly divergent PSSM scores are present in both plasma and PBMC, but a shift toward a more abundant representation of X4-like PSSM scores in PBMC derived DNA was apparent. Additional studies are needed to evaluate the clinical importance of these findings with regard to tropism prediction and the use of CCR5 antagonists. PMID- 19131902 TI - Community-acquired bacteremia among children admitted to a rural hospital in Mozambique. AB - BACKGROUND: Although community-acquired bacteremia is an important cause of childhood mortality in Africa, recognition of disease burden and potential impact of bacterial vaccines is limited. METHODS: Blood cultures for bacterial pathogens were conducted systematically among children <15 years of age admitted to Manhica District Hospital, from 2001 to 2006. RESULTS: Blood-stream infections were identified in 8% (1550/19,896) of pediatric hospital admissions. Nontyphoidal Salmonella (NTS) and Pneumococcus were the most prevalent pathogens isolated (26% and 25% of 1550 cases, respectively). Until 28 days of life, Staphylococcus aureus (39%) and group B Streptococcus (20%) predominated. Incidence of community acquired bacteremia per 100,000 child-years was 1730/10 in children <1 year old, 782/10 in 1-4 year oldd, and 49/10 in children 5 years and older. Case-fatality of bacteremia was 12%. Community-acquired bacteremia associated mortality accounted for 21% (162/788) of hospital deaths. Resistance to antibiotics commonly used in Mozambique was high among invasive isolates of Haemophilus influenzae, Escherichia coli, and NTS. CONCLUSIONS: Community-acquired bacteremia is an important cause of pediatric hospital admission and death in rural African hospitals. The high burden of disease, mortality, and pattern of antibiotic resistance associated with bacteremia underscore the need for prevention in Sub Saharan Africa. PMID- 19131905 TI - Correctional health care must be recognized as an integral part of the public health sector. PMID- 19131904 TI - Orphanhood predicts delayed access to care in Ugandan children. AB - We aimed to determine patient characteristics, treatment challenges, and adherence outcomes according to orphanhood status in a prospective cohort of 101 HIV+ children in rural Uganda. Orphans were older at antiretroviral initiation (P = 0.0008) and more likely to be WHO stage-4 (P = 0.03) than nonorphans. More attention to improving access to antiretrovirals in pediatric populations, especially orphans, is needed. PMID- 19131903 TI - Demonstration of immunologic memory using serogroup C meningococcal glycoconjugate vaccine. AB - BACKGROUND: Studies of glycoconjugate vaccines have traditionally used an immune challenge with a plain polysaccharide vaccine to demonstrate immunologic memory. Plain polysaccharide vaccines are poorly immunogenic in children and can induce subsequent immunologic hyporesponsiveness. We therefore assessed the use of glycoconjugate vaccines as an alternative method of demonstrating immunologic memory. METHODS: Children immunized with hepatitis B vaccine or serogroup C meningococcal glycoconjugate vaccine (MenCC) at age 2, 3, 4 months received a plain polysaccharide meningococcal serogroup A/C vaccine (MenACP) or MenCC at age 12 months. A post hoc analysis of serum bactericidal activity responses to MenCC assessed whether this differed in MenCC primed and MenCC naive infants. RESULTS: MenCC primed children displayed higher geometric mean serum bactericidal titers than MenCC naive children following MenACP (1518 compared with 30; P = 0.003). A similar difference was seen after a dose of MenCC to toddlers (MenCC primed: 8663, MenCC naive: 710; P < 0.001). The latter comparison became a borderline significance after adjusting for higher pretoddler immunization serum bactericidal geometric mean titers in the MenCC primed group (P = 0.068). CONCLUSIONS: Administration of glycoconjugate vaccines provides an important alternative method of demonstrating immunologic memory, avoiding the use of plain polysaccharide vaccines that are potentially deleterious in children. This has implications for the design of all future clinical trials of glycoconjugate vaccines. PMID- 19131906 TI - An evaluation of HIV partner counseling and referral services using new disposition codes. AB - BACKGROUND: New Centers for Disease Control recommendations suggest that all persons with newly diagnosed HIV receive partner counseling and referral services (PCRS). METHODS: We evaluated the King County, WA, PCRS program using a new set of disposition codes that disaggregate the components of PCRS (notification, testing, and test results), distinguish verified and unverified outcomes, and differentiate outcomes that occur before and after cases receive PCRS. RESULTS: Between 2005 and 2007, 427 (65%) of 659 persons with newly diagnosed HIV received PCRS. The number of cases staff needed to interview to identify 1 new case of HIV varied from 12.2 to 47.4 depending on whether number needed to interview was defined to include both verified and unverified outcomes and whether it excluded partners diagnosed with HIV before cases' receipt of PCRS. Age <25, testing HIV negative within the last year, receipt of PCRS within 58 days of HIV diagnoses, and participation in a program to link persons with HIV to medical care were significantly associated notifying more partners. CONCLUSIONS: PCRS evaluations may overestimate success because of limitations inherent in Centers for Disease Control PCRS disposition codes. Efforts to promote frequent HIV testing, assure timely provision of PCRS, and integrate PCRS with programs that link patients to care may improve PCRS outcomes. PMID- 19131907 TI - Self-medication prevalence for sexually transmitted diseases: meta-analysis and meta-regression of population level determinants. AB - BACKGROUND: Estimated prevalence of self-medication for sexually transmitted diseases (STDs) in observational studies differs between studies and it may vary according to population characteristics. METHODS: We investigated the extent of self-medication use for STD and explored population and study level factors correlated to the variability observed between populations. Medical databases were systematically searched for published studies. Crude prevalence data were extracted. Pooled estimates were calculated using random effects models. Univariate and multivariate meta-regression models with categorical population level covariates were constructed. RESULTS: Of the 35 studies that met the inclusion criteria for all self-medication use, 20 were also included in the analysis of antibiotic only use. There was significant heterogeneity present across prevalence estimates for all self-medication, Q = 3954.82, P <0.001; and among antibiotic only self-medication, Q = 3797.94, P <0.001. In meta-regression analyses, publication year seemed to explain 22.5% of variation among studies of self-medication with antibiotic only. No other significant associations with population or study characteristics were found. CONCLUSIONS: No population level factors leading to high levels of self-medication use for STD were identified. Publications before year 2000 seem to present a higher prevalence of antibiotic use for self-medication. Greater risk awareness of antibiotic misuse and improved control of antibiotic availability at population level might explain this result. An analysis of individual level characteristics should indicate which are influential and whether their local prevalence can explain the observed heterogeneity better. PMID- 19131908 TI - Prevalence and factors associated with human immunodeficiency virus infection among sex workers in Samarkand, Uzbekistan. PMID- 19131909 TI - The impact of discontinuation of male STD screening services at a large urban county jail: Chicago, 2002-2004. AB - BACKGROUND: Before April 2003, all male detainees were offered chlamydia (CT) and gonorrhea (GC) screening tests, after which services were limited to symptom based testing. In 2003, male screening was discontinued at a large urban county jail. OBJECTIVE: To evaluate the impact of discontinuing universal male sexually transmitted disease screening in a large county jail. METHODS: We compared the number of male CT/GC cases during the periods of universal screening (April 2002 to March 2003) with symptom-based testing (April 2003 to March 2004). RESULTS: The number of reported CT/GC cases among male detainees declined by -91.7% (3329 277) and -90.5% (1133-108), respectively after universal screening was discontinued. Citywide, CT/GC cases among males and females declined by -9.3% (24,885-22,563) and -12.9% (13,249-11,541), respectively. CONCLUSIONS: Discontinuation of universal male CT/GC screening services at a large county jail represents a missed opportunity to screen a high-risk population and was associated with substantial declines in reported morbidity. PMID- 19131911 TI - A prospective clinical audit of a new dressing design for lower limb arthroplasty wounds. AB - An audit of wound healing in patients undergoing total hip or knee arthroplasty in a Scottish hospital found that use of a barrier film made no difference to the effectiveness of a new Hydrofiber/hydrocolloid dressing combination. PMID- 19131912 TI - Current thoughts on angiogenesis. AB - Research into the distinct phases of the process of angiogenesis has informed the development of growth factor therapy and tissue-engineered products, but more studies are needed to ensure the development of new therapeutic strategies. PMID- 19131914 TI - Is TNP in the open abdomen safe and effective? AB - Much of the evidence for the use of TNP in the open abdomen comes from data on trauma patients. In view of the potentially severe complications, much greater evidence is needed for its application on patients with abdominal sepsis. PMID- 19131913 TI - The development of the Glamorgan paediatric pressure ulcer risk assessment scale. AB - OBJECTIVE: To develop a predictive pressure ulcer risk assessment scale based on patient data. METHOD: Detailed questionnaires, based on a review of the paediatric and adult literature and discussion with paediatric nurses with expertise in this area, were used to obtain data on patient characteristics from 265 inpatients in a children's hospital and 54 children with pressure ulcers and 17 children without pressure ulcers from 11 hospitals. These data were then combined to compare the characteristics of children with and without pressure ulcers (using the chi-square test). RESULTS: The Glamorgan paediatric pressure ulcer risk assessment scale was developed, based on the significance values identified by the statistical analysis. Variables that were significantly (p<0.01) associated with pressure ulceration included: difficult to position, anaemia, equipment pressing or rubbing against skin, reduced mobility for age, prolonged surgery and persistent pyrexia. The sensitivity, specificity and predictive validity of the Glamorgan scale was found to be greater than those for the Braden Q scale, which is widely used to assess pressure ulcer risk in children in the US. CONCLUSION: Using patient data from children and young people in the development of risk assessment tools for this age group may be preferable to modifying existing adult tools. PMID- 19131915 TI - Clinical experiences of using a cellulose dressing on burns and donor site wounds. AB - These preliminary results suggest this temporary skin substitute reduces pain and achieves comparable healing rates to other dressings. Furthermore, its transparency enables regular inspection of the wound bed for signs of infection. PMID- 19131916 TI - The impact of topical phenytoin on recalcitrant neuropathic diabetic foot ulceration. AB - OBJECTIVE: To study the impact of topical phenytoin on the healing of recalcitrant neuropathic diabetic foot ulcers in patients with no clinical evidence of ischaemia or infection, and to evaluate its antibacterial effect. METHOD: Thirty-two patients were enrolled into the study. Topical phenytoin in the form of 2% aerosol powder was applied once daily in addition to the patient's usual treatment (weekly sharp debridement, offloading and use of a gauze dressing) for eight weeks. The primary outcome was change in ulcer area over time, measured by grid tracing. Secondary outcomes were the ability of topical phenytoin to eradicate bacterial isolates, and the occurrence of adverse events. RESULTS: Topical phenytoin significantly improved healing of recalcitrant neuropathic diabetic foot ulcers. Baseline wound area was 319.3 + 340.4 mm2, reducing to 286.1 + 341.1 mm2 and 269.1 + 341.2 mm2 after four and eight weeks respectively. However, the overall reduction in ulcer size was only 18.3% + 27.5% and 25.7% + 38.6 % respectively. Topical phenytoin therapy over eight weeks did not eradicate any of the bacterial wound isolates (Staphylococcus spp., Proteus spp. or Pseudomonas spp.). Of the 32 patients evaluated, only eight (25%) achieved more than 50% reduction in ulcer size after eight weeks of treatment. CONCLUSION: Topical phenytoin can enhance wound healing in recalcitrant neuropathic diabetic foot ulcers, although only one-quarter of patients achieved more than 50% reduction in ulcer size after eight weeks of therapy. Further research is needed to characterise those patients who will satisfactorily respond to such therapy. PMID- 19131917 TI - Short-stretch bandages can assist wound healing and reduce oedema in immobile patients. PMID- 19131918 TI - An 18th century account of leg ulcer care that speaks to us today. AB - In 1766, a French physician proposed a protocol of care for patients with leg ulcers that anticipates much modern thinking on this topic. So, has our understanding increased as much as we think or is there still much to learn and do? PMID- 19131919 TI - One-stop hybrid approach for cardiovascular disease: from conception to practice. PMID- 19131920 TI - Early Barrett's carcinoma of the esophagus. PMID- 19131921 TI - In vitro-chemosensitivity test using the collagen gel droplet embedded culture drug test (CD-DST) for malignant pleural mesothelioma: possibility of clinical application. AB - PURPOSE: An in vitro-chemosensitivity test using the collagen gel droplet embedded culture drug test (CD-DST), established by Kobayashi et al. (Jpn J Cancer Res 2001; 92: 203-10), has been widely used on various tumors. This study retrospectively evaluated its possibility of clinical application to patients with malignant pleural mesothelioma (MPM). PATIENTS AND METHODS: CD-DST using 26 fresh specimens obtained by biopsy or surgery on MPM patients investigated in vitro responses to cisplatin (CDDP), carboplatin (CBDCA), doxorubicin (ADR), etoposide (VP-16), 5-fluoruracil (5-FU), gemcitabine (GEM), vinorelbine (VNR), irinotecan (SN-38), and docetaxel (TXT). Correlations between CD-DST data and clinical effects were then assessed for some MPM patients undergoing chemotherapy. RESULTS: The rate of in vitro sensitivity to each chemoagent (N=tested number) was 35% for CDDP (N=23), 14% for CBDCA (N=21), 7% for ADR (N=15), 15% for VP-16 (N=13), 0% for 5-FU (N=15), 45% for GEM (N=11), 25% for VNR (N=8), 40% for SN-38 (N=5), and 44% for TXT (N=9). No difference was observed between CD-DST data of each chemoagent and histological type. Of these MPM patients, 14 clinical effects on 13 patients who underwent chemotherapy for primary or recurrent disease were reviewed in comparison with CD-DST data of each chemoagent. Among 10 chemotherapies including in vitro-sensitive chemoagents, 3 led to partial response (PR), and 7 resulted in four stable diseases (SDs) and 3 to progressive diseases (PDs). In contrast, among 4 chemotherapies using in vitro resistant chemoagents, SD and PD were observed in 1 and 3, respectively. In regard to the clinical response rate, CD-DST sensitivity, specificity, and accuracy in the 14 examined chemotherapies were respectively 100%, 36%, and 50%, and in regard to the disease control rate, they were 88%, 60%, and 71%. CD-DST data for the chemoagents were to a limited extent significantly correlated with the disease control status of chemotherapy (p=0.052). CONCLUSION: Although the number of tested MPM specimens was small, CD-DST data obtained by biopsy or surgical-fresh specimens of MPM marginally correlated to the disease control effect of chemotherapy for this disease. Therefore CD-DST may possibly be applied to selecting the chemotherapy regimen for MPM. To determine the possibility of a clinical application of this test to MPM, a prospective clinical study of a greater number of patients will be necessary. PMID- 19131922 TI - Experimental study of the relationship between perfluoro-octyl bromide emulsion and norepinephrine release in reperfusion arrhythmia: isolated guinea pig heart model. AB - BACKGROUND: Perfluoro-octyl bromide (PFOB), one of the perfluorochemical oxygen transporters, improved postischemic cardiac dysfunctions. Also norepinephrine (NE) is one of the important inducible factors on reperfusion arrhythmias (ventricular fibrillation [VF]). We used these methods to evaluate the relationship between PFOB emulsion and NE release on reperfusion arrhythmias. MATERIALS AND METHODS: The perfusion of isolated guinea pig hearts was employed: each of four groups of 6-7 hearts were used with Krebs-Henseleit solution (KHS) as control, and KHS with 5%, 15%, or 30% PFOB emulsion. The hearts were perfused in a constant pressure Langendorff model, stabilized for 30 min, followed by 30 min preischemia, then 30 min ischemia and 45 min reperfusion at normothermia. RESULTS: PFOB emulsion dose-dependently limited VF and inhibited NE release in reperfusion. Only 30% PFOB emulsion showed the significant improvement of VF (p=0.05). In hemodynamic parameters, only 5% PFOB emulsion showed a significant decrease in reperfusion, but there was no difference in coronary flow rate (CFR). No differences among the four groups were demonstrated in cardiac oxygen metabolic parameters. CONCLUSIONS: It was most likely that a high concentration of PFOB emulsion attenuated reperfusion arrhythmia by decreasing NE release. PMID- 19131923 TI - Impact of obesity on early outcomes after cardiac surgery: experience in a Saudi Arabian center. AB - PURPOSE: The prevalence of obesity is a public health concern in most countries, including Saudi Arabia. Obesity has been considered a major risk factor for adverse outcomes after cardiac surgery. MATERIALS AND METHODS: A single-center retrospective review (2001-2005) of adverse outcome after coronary artery bypass grafting (CABG) and valve surgery (total=462) categorized by body mass index (BMI) was performed. The patients with BMI>or=30 were defined as the obese group and patients whose BMI<30 were labeled as the nonobese group. RESULTS: Overall, 315 (68.2%) were classified as nonobese, and 147 (31.8%) were obese. Obese patients were older and more likely to have diabetes and hypertension. There were no significant differences between the two groups with regard to other comorbidity and risk factors. There was no association between the two groups and the outcomes of operative mortality and morbidities. CONCLUSION: This study demonstrated that obesity does not increase the risk of death and most complications after cardiac surgery, aside from the unexplained increased risk of reoperation during the same admission. PMID- 19131924 TI - Near-infrared spectrophotometry is useful to detect the beneficial pharmacological effects of alprostadil on spinal cord deoxygenation. AB - PURPOSE: The purpose of this study is to confirm our previous studies that near infrared spectrophotometry (NIRS) can detect spinal cord ischemia earlier than evoked spinal cord potential (ESP), and to determine whether it can detect the pharmacological effect of prostaglandin E1 (PGE1) incorporated in lipid microspheres (alprostadil) on the spinal cord. MATERIALS AND METHODS: NIRS probes were placed on the posterior side of the lower lumbar vertebrae, and oxygenated hemoglobin (oxy-Hb) of the spinal cord was monitored continuously in 14 male New Zealand white rabbits. The amplitude of ESP was recorded every minute. All rabbits underwent a normothermic infrarenal aortic cross-clamping (AXC) for 20 min, and all were pretreated with either an intravenous 3 microg/kg/10 min alprostadil (group A; n=4) or the same volume saline (group C; n=10). RESULTS: ESP amplitude started to show a linear decrease 6 min after the onset of AXC and was comparable between groups (P=.839). Oxy-Hb decreased rapidly just after the onset of AXC, followed by monoexponential decline. It reached a plateau at 10 min after the onset of AXC. Oxy-Hb of group A was significantly higher than that of group C (P=.014). CONCLUSIONS: NIRS can detect spinal cord ischemia earlier than ESP. It can detect the beneficial pharmacological effect of alprostadil on the spinal cord. PMID- 19131925 TI - A case report: thoracic extramedullary hematopoiesis found by occurring spontaneous pneumothorax. AB - Extramedullary hematopoiesis (EMH) is a rare disease associated with hematologic disorders. This report describes a case of posterior mediastinal mass found by occurring spontaneous pneumothorax in a 48-year-old male. The intrathoracic mass resected using video-assisted thoracic surgery (VATS) was diagnosed thoracic EMH (TEMH). No disorders were found by hematologic exams before or after surgery. This report suggests that such lesions must therefore be considered in the differential diagnosis of posterior mediastinal tumors presenting with no hematologic disorders. PMID- 19131926 TI - Late pulmonary metastasis of a rectal cancer resected 20 years after low anterior resection. AB - A 54-year-old man underwent a low anterior resection for rectal cancer in July 1986. A right pulmonary tumor was pointed out in March 2006 by screening a chest X-ray. Because pulmonary metastasis from colorectal cancer was suspected by transbronchial biopsy, a colonofiberscopy was performed. However, no primary tumor was found. Furthermore, positron emission tomography (PET) revealed no other positive lesion. This tumor was resected in April 2006, and the pathological findings, including immunohistochemical findings, were similar to those of the previously resected rectal cancer. This tumor was therefore diagnosed to be pulmonary metastasis from rectal cancer resected 20 years ago based on a comparison of primary and pulmonary tumors using immunohistochemical examination. This shows that pulmonary metastasis from colorectal cancer is possible even 20 years after the initial operation. PMID- 19131927 TI - One-and-a-half ventricular repair through the right lateral thoracotomy: an alternative to midline approach in a patient with previous mediastinitis. AB - We report a patient who successfully underwent a one-and-a-half ventricular repair (1.5 VR) through a right lateral thoracotomy. In the case of possible hazardous complications at the sternal reentry because of previous mediastinitis, this approach was thought to be an option in selected patients to complete a functional correction by means of 1.5 VR. PMID- 19131928 TI - Surgical resection of cardiac papillary fibroelastoma in the left ventricular outflow tract. AB - An 81-year-old man was referred to our hospital on May 29, 2006, for a left ventricular tumor that a cardiologist had evaluated by echocardiography. The patient underwent surgical resection of a left ventricular tumor including the stalk through the aortic annulus on June 6. The aortic cross-clamp time, cardiopulmonary time, and operation time were 27 minutes, 48 minutes, and 2 hours 40 minutes, respectively. The specimen was examined pathologically and found to be a cardiac papillary fibroelastoma (CPF), 20x15 mm in size. He was discharged home 10 days after the surgery without significant adverse events. There are approximately 6 case reports on CPF in the left ventricular cavity published in Japan. Our surgical experience was reported, and a clinical feature of papillary fibroelastoma on the prior clinical data was reviewed retrospectively. PMID- 19131929 TI - Primary malignant pericardial mesothelioma presenting as pericardial constriction. AB - A 55-year-old man with a history of pericardiocentesis for massive pericardial effusion of unknown etiology was admitted to our hospital because of shortness of breath and systemic edema in September 2005. Transthoracic echocardiography demonstrated the massive PE 2 cm in diameter and with several areas of thick hyperrefractile echoes arising from the pericardium. Computed tomography (CT) demonstrated a large mediastinal mass encasing the heart; a pressure of the right ventricle (RV) showed a pattern of dips and plateaus on cardiac catheterization. Pericardiocentesis was attempted, but no fluid could be aspirated. The patient's symptoms progressed day by day despite maximum pharmacological support with catecholamines and diuretics. Surgical treatment was planned to relieve the symptoms and confirm the definitive diagnosis. Pericardiectomy and partial resection of the tumor under cardiopulmonary bypass (CPB) could be performed, and this resulted in a marked relief of symptoms. Histological examination confirmed the malignant pericardial mesothelioma. In conclusion, pericardiectomy and resection of the tumor might be indicated for the relief of symptoms in a critical case presenting as pericardial constriction associated with malignant pericardial mesothelioma. PMID- 19131930 TI - Aneurysm of the main pulmonary artery. AB - An aneurysm of the main pulmonary artery is a rare entity with differing management strategies found among the medical and surgical literature. The etiology of this entity is variable and its natural history unknown. We describe repair in an adult by means of graft interposition using cardiopulmonary bypass. We give our rationalization for surgical repair and present the contrary view of conservative management. PMID- 19131931 TI - Total arch replacement with endarterectomy of the ascending aorta in a patient with aortic arch aneurysm and porcelain aorta. AB - We describe total arch replacement for aortic arch aneurysm with a severely calcified ("porcelain") aorta. Cardiovascular surgery is challenging under such conditions because the calcified plate interferes with clamping, incising, and suturing of the aorta. We performed this surgery under hypothermic circulatory arrest with antegrade cerebral perfusion. Calcification manifested particularly in the ascending aorta and prevented the use of a needle. We exfoliated the calcified intimal plate using an elevator designed for hand surgery, and then covered both the inner and outer sides of the endarterectomized aorta with a strip of bovine pericardium to reinforce the anastomotic region and cover the rough surface. No complications developed during or after surgery. PMID- 19131932 TI - Recurrent coarctation of the aorta: a patient with bilateral persistent sciatic arteries. AB - A 19-year-old female was referred to our department with a diagnosis of recurrent coarctation of the aorta. She had undergone end-to-side anastomosis of the left subclavian artery with the thoracic aorta at 9 days of age and extra-anatomic subclavian-to-femoral bypass grafting at 15 years of age because of hypertensive crisis. A preoperative 4-slice computed tomography (CT) scan demonstrated recurrent obstruction of the aorta and, unexpectedly, an associated anomaly of bilateral persistent sciatic arteries. She underwent a successful patch augmentation of the aorta and is currently leading a normal life without need for antihypertensive medication. Persistent sciatic artery is a rare congenital vascular anomaly and is known to develop aneurysm or critical lower limb ischemia. To our knowledge, this is the first report of a patient with coarctation of the aorta and bilateral persistent sciatic arteries. PMID- 19131933 TI - Association of antihypertensive monotherapy with serum sodium and potassium levels in Chinese patients. AB - BACKGROUND: International guidelines on management of hypertension recommend any major classes of antihypertensive drugs. However, the low prescribing rate of thiazides has been attributed to concerns about electrolyte disturbances and studies between antihypertensive drug classes and hyponatremia/hypokalemia among Chinese patients were scarce. METHODS: From clinical databases we included 2,759 patients who received their first-ever antihypertensive monotherapy from January 2004 to June 2007 in a large territory of Hong Kong. We studied the plasma sodium and potassium levels 8 weeks after prescriptions and factors associated with hyponatremia and hypokalemia by multivariable regression analyses. RESULTS: Among major antihypertensive drug classes, thiazide users had the lowest sodium level (139.6 mEq/l, 95% confidence interval (CI) 139.3, 140.0, P < 0.001) and patients prescribed calcium channel blockers (CCBs; 3.92 mEq/l, 95% CI 3.89, 3.95) or thiazide diuretics (3.99 mEq/l, 95% CI 3.93, 4.04) had the lowest potassium levels (P < 0.001). Multivariate analysis reported that advanced age (> or =70 years, odds ratio (OR) 7.49, 95% CI 2.84, 19.8, P < 0.001), male gender (OR 2.38, 95% CI 1.45, 3.91, P < 0.001), and thiazide users (OR 2.42, 95% CI 1.29, 4.56, P = 0.006) were significantly associated with hyponatremia, while renin-angiotensin system (RAS) (OR 0.31, 95% CI 0.13, 0.73, P = 0.008) and beta-blockers (BBs) (OR 0.35, 95% CI 0.23, 0.54, P < 0.001) users were less likely to present with hypokalemia. However, the proportions having normonatremic (95.1%) and normokalemic (89.4%) levels were high. CONCLUSIONS: In view of the low prevalence of hyponatremia and hypokalemia associated with thiazides, physicians should not be deterred from prescribing thiazide diuretics as first-line antihypertensive agents as recommended by most international guidelines. PMID- 19131934 TI - Podocyte loss in human hypertensive nephrosclerosis. AB - BACKGROUND: Podocyte injury probably plays important roles in the pathogenesis of hypertensive nephropathy, but human data are limited. We studied glomerular podocyte count and intrarenal expression of podocyte-associated molecules in patients with hypertensive nephrosclerosis. METHODS: We studied 41 consecutive patients with biopsy-proven hypertensive nephropathy, 10 cadaveric kidney donors, and 9 healthy subjects. Intrarenal and urinary mRNA expression of podocyte associated molecules was quantified and podocyte number was determined by stereological method. RESULTS: Intrarenal mRNA expression levels of nephrin, podocin, and synaptopodin were lower, and urinary mRNA expression levels were higher in patients with hypertensive nephropathy than controls. Glomerular podocyte number was significantly lower in patients with hypertensive nephrosclerosis than kidney donors (545 +/- 237 vs. 773 +/- 296 per glomeruli, P < 0.02). Podocyte density and intrarenal gene expression of podocyte-associated molecules significantly correlated with estimated glomerular filtration rate (GFR) and inversely correlated with blood pressure and the degree of renal fibrosis. Intrarenal gene expression of nephrin significantly correlated with change in renal function decline. CONCLUSION: This study demonstrates a decrease in podocyte number and intrarenal gene expression of podocyte-associated molecules in patients with hypertensive nephrosclerosis. We observed correlations between glomerular podocyte density and intrarenal expression of podocyte associated molecules with renal function and the degree of renal fibrosis, suggesting podocyte loss may play an important role in the pathogenesis of hypertensive nephrosclerosis. PMID- 19131935 TI - Association of metabolic syndrome and blood pressure nondipping profile in untreated hypertension. AB - BACKGROUND: There is a marked association between metabolic syndrome (MS) and increased cardiovascular risk. Moreover, nondipping (patients with <10% decline in the asleep relative to the awake blood pressure (BP) mean) has also been associated with increased cardiovascular morbidity and mortality. METHODS: We investigated the association between MS and impaired nocturnal BP decline in 1,770 nondiabetic, untreated hypertensive patients (824 men and 946 women), 48.7 +/- 13.2 years of age. BP was measured by ambulatory monitoring for 48 h to increase reproducibility of the dipping pattern. Physical activity was simultaneously monitored every minute by wrist actigraphy. RESULTS: MS was present in 42.4% of the patients. The prevalence of a nondipper BP profile was significantly higher in patients with MS (46.1% vs. 37.5% in patients without MS, P < 0.001). Patients with MS were characterized by significant elevations in uric acid (5.9 mg/dl vs. 5.2 mg/dl, P < 0.001), fibrinogen (314 mg/dl vs. 304 mg/dl, P = 0.021), and globular sedimentation rate (13.8 mm vs. 11.6 mm, P < 0.001). Nondipping was significantly associated to the presence of MS in a multiple logistic regression model adjusted by other significant confounding factors, including age, serum creatinine, and cigarette smoking. The single most relevant factor in the definition of MS associated to nondipping was elevated waist perimeter. CONCLUSIONS: This study documents a significant increase of a blunted nocturnal BP decline in patients with MS. Patients with MS were also characterized by elevated values of relevant markers of cardiovascular risk, including fibrinogen and globular sedimentation rate. PMID- 19131936 TI - Association of (pro)renin receptor gene polymorphism with blood pressure in Japanese men: the Ohasama study. AB - BACKGROUND: Recent studies have revealed that (pro)renin receptor ((P)RR), a newly identified member of the renin-angiotensin system (RAS), is associated with blood pressure regulation in animals. However, there is no information on (P)RR in humans. We investigated the association of (P)RR gene polymorphisms with blood pressure in a Japanese population. METHODS: Subjects (n = 1,112) were recruited from participants in the Ohasama study, a Japanese cohort study. For the association study, we selected three polymorphisms: -782A>G (rs2968915), intervening sequence (IVS)5+169C>T (rs5918007), and +1513A>G (rs6609080). Because the (P)RR gene is on the X chromosome, men (n = 357) and women (n = 755) were analyzed separately. RESULTS: In men, 24-h systolic blood pressure (SBP) and diastolic blood pressure (DBP) values, daytime SBP and DBP values, and nighttime SBP and DBP values were significantly higher in IVS5+169T allele carriers than C allele carriers. Multiple regression analysis showed that IVS5+169C>T was significantly and independently related to ambulatory blood pressure (ABP). IVS5+169C>T was not associated with casual blood pressure (CBP) in men. In women, there were no significant differences in blood pressure values among the three genotypes of IVS5+169C>T. This polymorphism had no significant association with any other clinical characteristic. -782A>G was weakly associated with ABP in men. +1513A>G was not associated with blood pressure values in either men or women. CONCLUSIONS: We demonstrated for the first time that polymorphism of the (P)RR gene IVS5+169C>T is associated with ABP in Japanese men. This association suggests that (P)RR has a role in blood pressure regulation. PMID- 19131937 TI - Managing childhood overweight: behavior, family, pharmacology, and bariatric surgery interventions. PMID- 19131938 TI - Elderly people with low body weight may have subtle low-grade inflammation. AB - Low-grade inflammation, which plays important roles in the development of fatal diseases, is commonly observed in obese people. However, this has not been evaluated in lean people, who have relatively increased mortality risk compared with people of normal weight. Here, we elucidate the association between systemic low-grade inflammation and low body weight, with particular emphasis on aging. We examined the relationship between circulating C-reactive protein (CRP) and BMI in a cross-sectional study of 2,675 apparently healthy adults who had undergone a medical check-up. Overall, subjects with low BMI (<21.0 kg/m(2), n = 585) showed a favorable cardiovascular profile without being undernourished. In the elderly (>or=55 years old), logarithmic CRP (LogCRP) showed a sigmoid curve against BMI with a base at BMI 21.0-22.9 kg/m(2), but not against waist circumference (WC), even in nonsmokers. In contrast, in middle-aged people, LogCRP showed an almost linear relationship with both BMI and WC. LogCRP levels in elderly nonsmokers with low BMI, but not normal or high BMI, were significantly higher than those in middle-aged with corresponding BMI (P < 0.05). After adjustment for age, sex, smoking status, and weight change over the past 2 years, the adjusted means of LogCRP still had a similar sigmoid curve against BMI in the elderly. These results suggest that elderly people with low body weight may have subtle low grade inflammation irrespective of a favorable cardiovascular risk, which remains to be confirmed in further studies. PMID- 19131939 TI - Validation of DXA body composition estimates in obese men and women. AB - The aim of this study was to determine the accuracy of dual-energy X-ray absorptiometry (DXA)-derived percentage fat estimates in obese adults by using four-compartment (4C) values as criterion measures. Differences between methods were also investigated in relation to the influence of fat-free mass (FFM) hydration and various anthropometric measurements. Six women and eight men (age 22-54 years, BMI 28.7-39.9 kg/m(2), 4C percent body fat (%BF) 31.3-52.6%) had relative body fat (%BF) determined via DXA and a 4C method that incorporated measures of body density (BD), total body water (TBW), and bone mineral mass (BMM) via underwater weighing, deuterium dilution, and DXA, respectively. Anthropometric measurements were also undertaken: height, waist and gluteal girth, and anterior-posterior (A-P) chest depth. Values for both methods were significantly correlated (r(2) = 0.894) and no significant difference (P = 0.57) was detected between the means (DXA = 41.1%BF, 4C = 41.5%BF). The slope and intercept for the regression line were not significantly different (P > 0.05) from 1 and 0, respectively. Although both methods were significantly correlated, intraindividual differences between the methods were sizable (4C-DXA, range = 3.04 to 4.01%BF) and significantly correlated with tissue thickness (chest depth) or most surrogates of tissue thickness (body mass, BMI, waist girth) but not FFM hydration and gluteal girth. DXA provided cross-sectional %BF data for obese adults without bias. However, individual data are associated with large prediction errors (+/-4.2%BF). This error appears to be associated with tissue thickness indicating that the DXA device used may not be able to accurately account for beam hardening in obese cohorts. PMID- 19131940 TI - Independent association of HbA(1c) and incident cardiovascular disease in people without diabetes. AB - Recent studies have reported no association between elevated glycated hemoglobin (HbA(1c)) and incident cardiovascular disease (CVD) among women without diabetes. This study describes associations between HbA(1c) and new onset CVD in a representative adult population cohort. Assessment of participants in The North West Adelaide Health Study (NWAHS), a population study of randomly selected adults (age > or =18 years, n = 4,060), included measurement of height, weight, blood pressure, fasting lipids, glucose, and HbA(1c). A self-completed questionnaire assessed doctor-diagnosed diabetes, CVD and stroke, smoking status, and demographics. The cohort was followed for an average 3.5 years. Of the 2,913 adults free of diabetes at baseline and follow-up, 94 (3.5%) reported new onset coronary heart disease (CHD) and/or stroke. Compared with those with an HbA(1c) < or =5.0%, risk of new onset CVD was increased in those with HbA(1c) 5.4-5.6% (odds ratio (OR) 2.5, 95% confidence interval (CI) 1.4, 4.6), and > or =5.7% (OR 1.9, 95% CI 1.1, 3.4), after adjustment for other risk factors. The association was stronger in women than men (P = 0.03), and attenuated to only a small degree by addition of impaired fasting glucose (IFG), hypertension, hypercholesterolemia, BMI, waist circumference, or smoking to the model. Elevated HbA(1c) is related to new onset CVD over a relatively short follow-up period in both men and women without diabetes and who do not develop diabetes, after adjustment for other major risk factors. Unlike previous studies, this relationship was not substantially attenuated by other traditional risk factors. PMID- 19131941 TI - Anti-inflammatory effect of palmitoylethanolamide on human adipocytes. AB - Obesity leads to the appearance of an inflammatory process, which can be initiated even with a moderate weight gain. Palmitoylethanolamide (PEA) is an endogenous lipid, secreted by human adipocytes, that possesses numerous anti inflammatory properties. The main purpose of this study was to investigate the anti-inflammatory effect of PEA on human adipocytes, as well as in a murine model. The production of tumor necrosis factor-alpha (TNF-alpha) by lipopolysaccharide (LPS)-treated human subcutaneous adipocytes in primary culture and CF-1 mice was investigated by enzyme-linked immunosorbent assay. The effects of PEA on adipocyte TNF-alpha secretion were explored as well as some suspected PEA anti-inflammatory pathways: nuclear factor-kappaB (NF-kappaB) pathway, peroxisome proliferator-activated receptor-alpha (PPAR-alpha) gene expression, and TNF-alpha-converting enzyme (TACE) activity. The effects of PEA on the TNF alpha serum concentration in intraperitoneally LPS-treated mice were also studied. We demonstrate that the LPS induced secretion of TNF-alpha by human adipocytes is inhibited by PEA. This action is neither linked to a reduction in TNF-alpha gene transcription nor to the inhibition of TACE activity. Moreover, PPAR-alpha is not implicated in this anti-inflammatory activity. Lastly, PEA exhibits a wide-reaching anti-inflammatory action as the molecule is able to completely inhibit the strong increase in TNF-alpha levels in the serum of mice treated with high doses of LPS. In view of its virtual lack of toxicity, PEA might become a potentially interesting candidate molecule in the prevention of obesity-associated insulin resistance. PMID- 19131942 TI - Oxidant stress in healthy normal-weight, overweight, and obese individuals. AB - This study was undertaken to investigate the association among BMI and lipid hydroperoxide (LH), total antioxidant status (TAS), superoxide dismutase (SOD), and reduced glutathione (GSH). Ninety (n = 90) healthy males and females (n = 23/67) (29 normal weight (BMI: 22.74 +/- 0.25 kg/m(2)), 36 overweight (BMI: 27.18 +/- 0.23 kg/m(2)), and 25 obese (33.78 +/- 0.48 kg/m(2))) participated in the study. Data collected included anthropometric measures, fasting blood glucose, lipid profile, LH, TAS, and enzymatic antioxidants (SOD, and reduced GSH). The results of the study showed that obese individuals had significantly increased LH levels compared to normal-weight individuals (obese vs. normal weight (0.88 +/- 0.05 vs. 0.67 +/- 0.03 micromol/l, P < 0.01)) but the increased levels were not significantly different when compared to the overweight group (obese vs. overweight (0.88 +/- 0.05 vs. 0.79 +/- 0.05 micromol/l)). No other consistent significant differences in TAS, SOD, and GSH were identified between groups. This study concluded that only obesity and not moderate overweight elevates LH levels. Furthermore, the levels of TAS, SOD, and GSH in obesity do not explain the increased LH levels observed in obesity. PMID- 19131943 TI - Pregravid weight is associated with prior dietary restraint and psychosocial factors during pregnancy. AB - The objective of this study was to investigate the association of pregravid weight status, previous dietary restraint, and psychosocial factors during pregnancy. We used data from the Pregnancy, Infection, and Nutrition study, which recruited 2,006 women at prenatal clinics before 20 weeks' gestation who were >16 years and English speaking. Institute of Medicine BMI cut points of underweight (<19.8), normal weight (19.8-26.0), overweight (>26.0-29.0), obese (>29.0-34.9), and an additional category extremely obese (> or =35.0), were used to categorize weight status. Eight psychosocial measures and dietary restraint were assessed with regard to BMI; perceived stress, trait anxiety, depressive symptoms, internal locus of control (LOC), chance LOC, powerful others LOC, self-esteem, and mastery. Linear regression was used to estimate associations, controlling for potential confounders. A significant trend was found between increasing pregravid weight categories and increasing scores for perceived stress, trait anxiety, depressive symptoms, powerful others LOC and dietary restraint, and decreasing scores for self-esteem and mastery. In adjusted models, pregravid obesity was independently associated with perceived stress, trait anxiety, and depressive symptoms. Extreme obesity was independently associated with all measures except internal LOC. A strong linear association was found between increasing weight categories and dietary restraint. In conclusion, as pregravid weight increased, pregnant women were at greater risk of reporting higher scores on negative psychosocial state and dietary restraints, and reporting lower score on positive personal dispositions. PMID- 19131944 TI - Association between adult attention deficit/hyperactivity disorder and obesity in the US population. AB - Attention deficit hyperactivity disorder (ADHD) is a neurobehavioral disorder that affects approximately 2.9-4.7% of US adults. Studies have revealed high rates of ADHD (26-61%) in patients seeking weight loss treatment suggesting an association between ADHD and obesity. The objective of the present study was to test the association between ADHD and overweight and obesity in the US population. Cross-sectional data from the Collaborative Psychiatric Epidemiology Surveys were used. Participants were 6,735 US residents (63.9% white; 51.6% female) aged 18-44 years. A retrospective assessment of childhood ADHD and a self report assessment of adult ADHD were administered. Diagnosis was defined by three categories: never met diagnostic criteria, met full childhood criteria with no current symptoms, and met full childhood criteria with current symptoms. The prevalence of overweight and obesity was 33.9 and 29.4%, respectively, among adults with ADHD, and 28.8 and 21.6%, respectively, among persons with no history of ADHD. Adult ADHD was associated with greater likelihood of overweight, (odds ratio (OR) = 1.58; 95% confidence interval (CI) = 1.05, 2.38) and obesity (OR = 1.81; 95% CI = 1.14, 2.64). Results were similar when adjusting for demographic characteristics and depression. Mediation analyses suggest that binge eating disorder (BED), but not depression, partially mediates the associations between ADHD and both overweight and obesity. Results suggest that adult ADHD is associated with overweight and obesity. PMID- 19131945 TI - Epstein-Barr virus in cutaneous T-cell lymphomas: evaluation of the viral presence and significance in skin and peripheral blood. AB - The importance of viral agents in the development of cutaneous T-cell lymphomas (CTCL) is still debated. For this purpose, we retrospectively evaluated the Epstein-Barr virus (EBV) presence in Sezary syndrome (SS), mycosis fungoides (MF), inflammatory dermatoses (ID), and healthy donors (HD) using different approaches: EBV-DNA was quantified in skin biopsies and peripheral blood using real-time PCR, EBV-encoded small RNA (EBER) transcripts were detected by in situ hybridization (ISH), and latent membrane protein1-2 antigens were detected by immunohistochemistry. Skin biopsies were EBV-DNA-positive in 8/30 (27%) SS, 7/71 (10%) MF, and 2/18 (11%) ID patients and in none of the 25 normal skin samples. Positive mRNA (EBER) signals, always confined to cerebriform T lymphocytes, were found in 5/30 SS patients (17%), whereas signals in all MF and ID patients were negative. The presence of EBV-DNA in skin and blood samples was associated with a significantly lower survival in MF/SS patients. In evaluating EBV serological status, most (>70%) SS, MF, and ID patients showed a serological reactivation demonstrated by the presence of anti-EA IgG. In conclusion, although the finding of EBV-DNA in CTCL does not prove its etiopathogenetic role and may be related instead to immunosuppression, our study demonstrates that it has prognostic relevance. PMID- 19131947 TI - Independent evaluation of a commercial test for "autoimmune" urticaria in normal and chronic urticaria subjects. PMID- 19131946 TI - Increasing burden of melanoma in the United States. AB - It is controversial whether worldwide increases in melanoma incidence represent a true epidemic. Dramatic increases in incidence in the setting of relatively stable mortality trends have also been attributed to expanded skin screening and detection of biologically indolent tumors with low metastatic potential. To better understand how melanoma incidence trends varied by severity at diagnosis and factors relevant to screening access, we assessed recent United States incidence and mortality trends by histologic type, tumor thickness, and area level socioeconomic status (SES). We obtained population-based data regarding diagnoses of invasive melanoma among non-Hispanic whites from nearly 291 million person-years of observation by the Surveillance Epidemiology and End Results (SEER) program (1992-2004). Age-adjusted incidence and mortality rates were calculated for SEER and a subset (California) for which small-area SES measure was available. Overall, melanoma incidence increased at 3.1% (P<0.001) per year. Statistically significant rises occurred for tumors of all histologic subtypes and thicknesses, including those >4 mm. Melanoma incidence rates doubled in all SES groups over a 10-year period whereas melanoma mortality rates did not increase significantly. We conclude that screening-associated diagnosis of thinner melanomas cannot explain the increasing rates of thicker melanomas among low SES populations with poorer access to screening. PMID- 19131948 TI - Molecular analysis of 250 patients with autosomal recessive congenital ichthyosis: evidence for mutation hotspots in ALOXE3 and allelic heterogeneity in ALOX12B. AB - In recent years several new genes for autosomal recessive congenital ichthyosis (ARCI) have been identified. However, little is known about the molecular epidemiology and pathophysiology of this genetically and clinically heterogeneous group of severe disorders of keratinization. ARCI is characterized by intense scaling of the whole integument often associated with erythema. We and others have shown that mutations in ALOX12B and ALOXE3, coding for the lipoxygenases 12R LOX and eLOX-3 predominantly synthesized in the epidermis, can underlie this rare condition. Here we have surveyed a large group of 250 patients with ARCI for mutations in these two genes. We have identified 11 different previously unreported mutations in ALOX12B and ALOXE3 in 21 ARCI patients from 19 unrelated families and demonstrated that mutations in the two genes are the second most common cause for ARCI in this cohort of patients. Examination of the molecular data revealed allelic heterogeneity for ALOX12B and two mutational hotspots in ALOXE3. Functional analysis of all missense mutations and a splice site mutation demonstrated that complete loss of function of the enzymes underlies the phenotype. Our findings further establish the pivotal role of the 12-lipoxygenase pathway during epidermal differentiation. PMID- 19131949 TI - Polyclonality of multiple sporadic basal cell carcinomas. PMID- 19131950 TI - Single nucleotide polymorphism array analysis defines a specific genetic fingerprint for well-differentiated cutaneous SCCs. AB - Cutaneous squamous cell carcinomas (cSCCs) are the second most frequent cancers in fair-skinned populations; yet, because of their genetic heterogeneity, the key molecular events in cSCC tumorigenesis remain poorly defined. We have used single nucleotide polymorphism microarray analysis to examine genome-wide allelic imbalance in 60 cSCCs using paired non-tumor samples. The most frequent recurrent aberrations were loss of heterozygosity at 3p and 9p, observed in 39 (65%) and 45 (75%) tumors, respectively. Microdeletions at 9p23 within the protein tyrosine phosphatase receptor type D (PTPRD) locus were identified in 9 (15%) samples, supporting a tumor suppressor role for PTPRD in cSCC. In addition, microdeletions at 3p14.2 were detected in 3 (5%) cSCCs, implicating the fragile histidine triad (FHIT) gene as a possible target for inactivation. Statistical analysis revealed that well-differentiated cSCCs demonstrated significantly fewer aberrations than moderately and poorly differentiated cSCCs; yet, despite a lower rate of allelic imbalance, some specific aberrations were observed equally frequently in both groups. No correlation was established between the frequency of chromosomal aberrations and immune or human papillomavirus status. Our data suggest that well differentiated tumors are a genetically distinct subpopulation of cSCC. PMID- 19131952 TI - Separation of sedimentary micron-sized particles for palaeoceanography and calcareous nannoplankton biogeochemistry. AB - A protocol is described for separating sub-20 microm-sized particles contained in sedimentary rocks into size fractions. Geochemical data from manually isolated foraminifera are commonly used in the interpretation of marine palaeoenvironments; problems associated with the isolation of calcareous nannofossils hampers their geochemical exploitation. However, geochemistry performed on calcareous nannofossil monotaxic assemblages should provide more meaningful data sets than those generated from the highly heterogeneous bulk carbonate. This protocol is based on cascade filtering steps, using polycarbonate membranes with well-calibrated pores. Strong ultrasonic treatment can further be applied to selectively reduce the size of particles for greater enrichment. Obtained residues frequently comprise near-monotaxic nannofossil assemblages. The application of this technique, which can be achieved within less than 2 days, has provided distinct fractions of coccoliths, calcareous dinoflagellate shells and also diagenetic monocrystals. This protocol is designed for application in reconstructing the history of water-column physicochemistry and diagenesis. It also has the potential to provide insights into the biogeochemistry of calcareous nannoplankton, including vital effects. PMID- 19131951 TI - Protein structure homology modeling using SWISS-MODEL workspace. AB - Homology modeling aims to build three-dimensional protein structure models using experimentally determined structures of related family members as templates. SWISS-MODEL workspace is an integrated Web-based modeling expert system. For a given target protein, a library of experimental protein structures is searched to identify suitable templates. On the basis of a sequence alignment between the target protein and the template structure, a three-dimensional model for the target protein is generated. Model quality assessment tools are used to estimate the reliability of the resulting models. Homology modeling is currently the most accurate computational method to generate reliable structural models and is routinely used in many biological applications. Typically, the computational effort for a modeling project is less than 2 h. However, this does not include the time required for visualization and interpretation of the model, which may vary depending on personal experience working with protein structures. PMID- 19131953 TI - Dictyostelium discoideum: a model host to measure bacterial virulence. AB - Dictyostelium amoebae have been used as a host model to measure virulence of a wide range of bacterial pathogens. The simple protocol described here takes advantage of the ability of Dictyostelium to grow and form plaques on a lawn of nonpathogenic bacteria but not on virulent bacteria. This assay can be modulated to measure the virulence of different bacterial pathogens. By adjusting various parameters such as cell numbers or media, a more quantitative measure of bacterial virulence can also be obtained. The entire procedure takes about 5 h to compete, and up to 2 further weeks may be required for plaques to form on the bacterial lawn. PMID- 19131954 TI - Immunodesign of experimental sepsis by cecal ligation and puncture. AB - Sepsis remains a prevalent clinical challenge and the underlying pathophysiology is still poorly understood. To investigate the complex molecular mechanisms of sepsis, various animal models have been developed, the most frequently used being the cecal ligation and puncture (CLP) model in rodents. In this model, sepsis originates from a polymicrobial infectious focus within the abdominal cavity, followed by bacterial translocation into the blood compartment, which then triggers a systemic inflammatory response. A requirement of this model is that it is performed with high consistency to obtain reproducible results. Evidence is now emerging that the accompanying inflammatory response varies with the severity grade of sepsis, which is highly dependent on the extent of cecal ligation. In this protocol, we define standardized procedures for inducing sepsis in mice and rats by applying defined severity grades of sepsis through modulation of the position of cecal ligation. The CLP procedure can be performed in as little as 10 min for each animal by an experienced user, with additional time required for subsequent postoperative care and data collection. PMID- 19131955 TI - Northern blot analysis for detection and quantification of RNA in pancreatic cancer cells and tissues. AB - Investigation of gene expression significantly contributes to our knowledge of the regulation and function of genes in many areas of biology. In this protocol, we describe how northern blot analysis is used to identify gene expression patterns at the RNA level in human cancer cells as well as in cancerous and normal tissues. RNA molecules are separated by gel electrophoresis and are subsequently transferred to a porous membrane by capillary action. Specific sequences in the RNA are detected on the membrane by molecular hybridization with radiolabeled nucleic acid probes. Despite the development of newer methods, such as real-time PCR, nuclease protection assays and microarrays, northern blot analysis is still a standard technique used in the detection and quantification of mRNA levels because it allows a direct comparison of the mRNA abundance between samples on a single membrane. This entire northern blotting protocol takes approximately 4 d to complete. PMID- 19131956 TI - Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. AB - DAVID bioinformatics resources consists of an integrated biological knowledgebase and analytic tools aimed at systematically extracting biological meaning from large gene/protein lists. This protocol explains how to use DAVID, a high throughput and integrated data-mining environment, to analyze gene lists derived from high-throughput genomic experiments. The procedure first requires uploading a gene list containing any number of common gene identifiers followed by analysis using one or more text and pathway-mining tools such as gene functional classification, functional annotation chart or clustering and functional annotation table. By following this protocol, investigators are able to gain an in-depth understanding of the biological themes in lists of genes that are enriched in genome-scale studies. PMID- 19131957 TI - Engineering complex-type N-glycosylation in Pichia pastoris using GlycoSwitch technology. AB - Here we provide a protocol for engineering the N-glycosylation pathway of the yeast Pichia pastoris. The general strategy consists of the disruption of an endogenous glycosyltransferase gene (OCH1) and the stepwise introduction of heterologous glycosylation enzymes. Each engineering step results in the introduction of one glycosidase or glycosyltransferase activity into the Pichia endoplasmic reticulum or Golgi complex and consists of a number of stages: transformation with the appropriate GlycoSwitch vector, small-scale cultivation of a number of transformants, sugar analysis and heterologous protein expression analysis. If desired, the resulting clone can be further engineered by repeating the procedure with the next GlycoSwitch vector. Each engineering step takes approximately 3 weeks. The conversion of any wild-type Pichia strain into a strain that modifies its glycoproteins with Gal(2)GlcNAc(2)Man(3)GlcNAc(2)N glycans requires the introduction of five GlycoSwitch vectors. Three examples of the full engineering procedure are provided to illustrate the results that can be expected. PMID- 19131958 TI - Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. AB - Transient gene expression is an indispensable tool for studying functions of gene products. In the case of plants, transient introduction of genes by Agrobacterium infiltration is a method of choice for many species. However, this technique does not work efficiently in Arabidopsis leaf tissue, the most widely used model system for basic plant biology research. Here we present an optimized protocol for biolistic delivery of plasmid DNA into the epidermis of Arabidopsis leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol yields efficient and reproducible transient expression of diverse genes and is exemplified here for use in a functional assay of a transcription repressor and for the subcellular localization and cell-to-cell movement of plant viral movement protein. This protocol is suitable for studies of biological function and subcellular localization of the gene product of interest directly in planta by utilizing different types of activity-based assays. Using this procedure, the data are obtained after 2-4 d of work. PMID- 19131959 TI - Primary support cultures of hippocampal and substantia nigra neurons. AB - Primary cultures of rat and murine hippocampal neurons are widely used to reveal cellular mechanisms in neurobiology. Their use is limited, as culturing at low density is often not possible or is dependent on sophisticated methods. Here we present a novel method for culturing embryonic (E16.5) murine hippocampal neurons, using a spatially separated ring of cortical neurons for neurotrophic support. This method allows long-term cultures at a very low cell density, and therefore, the study of single embryo preparations and isolated neurons. This method has been adopted for neurons from the substantia nigra (E16.5), with support from a ring of striatal neurons. PMID- 19131960 TI - Orthotopic mouse lung transplantation as experimental methodology to study transplant and tumor biology. AB - Unlike transplantation of other solid organs, vascularized mouse lung transplantation has only recently been developed. In this protocol, we describe a detailed method for performing a vascularized and aerated mouse orthotopic lung transplant, which to date represents the most physiological mouse model of lung transplantation. The procedure is divided into two separate portions consisting of donor harvest followed by implantation using the cuff technique for bronchovascular anastomoses. After a training period spanning several months, the procedure can be successfully mastered and, in experienced hands, requires approximately 90 min to perform. After an initial learning curve, perioperative survival is close to 100%. As the donor hematopoietic cells in the transplanted lung are replaced by those of the host over time, thereby creating a 'chimeric lung,' this model represents a novel research tool for the study of transplantation biology as well as other disease processes, such as malignancies. PMID- 19131961 TI - Assaying dynamic cell-cell junctional communication using noninvasive and quantitative fluorescence imaging techniques: LAMP and infrared-LAMP. AB - This protocol describes a fluorescence imaging assay, local activation of molecular fluorescent probes (LAMP), for measuring rates of intercellular dye transfer across gap junction channels in intact living cells. The LAMP method consists of four steps: (i) loading cells with a cell-permeable and photo activatable fluorophore, NPE-HCCC2/AM (acetoxymethyl ester); (ii) locally photolyzing a caged coumarin in one cell of a coupled cell pair to release the parent fluorophore, HCCC2; (iii) imaging cell-cell transfer of HCCC2; and (iv) analyzing rates of dye diffusion. Compared with other methods available for measuring junctional coupling, the LAMP method offers a number of advantages, including noninvasiveness, ease of quantification of coupling strength, good temporal resolution and compatibility with multicolor imaging. Moreover, as the LAMP assay can be carried out multiple times in the same coupled cell pairs, changes in molecular permeability of connexin channels can be tracked by comparing rates of dye transfer. Finally, NPE-HCCC2 and HCCC2 have high two photon uncaging and excitation efficiency, respectively, enabling two-photon uncaging and imaging to be combined to examine cell coupling in three dimensions (infrared-LAMP assay). It takes roughly 3 h or 4 h to complete a LAMP or an infrared-LAMP assay, respectively. PMID- 19131962 TI - A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. AB - We explain a protocol for straightforward isolation and culture of mesenchymal stem cells (MSCs) from mouse bone marrow (BM) to supply researchers with a method that can be applied in cell biology and tissue engineering with minimal requirements. Our protocol is mainly on the basis of the frequent medium change in primary culture and diminishing the trypsinization time. Mouse mesenchymal stem cells are generally isolated from an aspirate of BM harvested from the tibia and femoral marrow compartments, then cultured in a medium with Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) for 3 h in a 37 degrees C-5% CO(2) incubator. Nonadherent cells are removed carefully after 3 h and fresh medium is replaced. When primary cultures become almost confluent, the culture is treated with 0.5 ml of 0.25% trypsin containing 0.02% ethylenediaminetetraacetic acid for 2 min at room temperature (25 degrees C). A purified population of MSCs can be obtained 3 weeks after the initiation of culture. PMID- 19131963 TI - A methodology for the combined in situ analyses of the precursor and mature forms of microRNAs and correlation with their putative targets. AB - There are relatively few protocols described for the in situ detection of microRNA (miRNA) and they often use cryostat sections, signal amplification and hybridization or washes of 50-60 degrees C. This protocol describes in situ miRNA detection that can be done in paraffin-embedded, formalin-fixed tissue. Detection of the miRNA precursors can be done by RT in situ PCR, which can theoretically detect one copy per cell. The key variable for the RT in situ PCR protocol is optimal protease digestion, which is then followed by overnight DNase digestion and target specific incorporation of the reported nucleotide into the amplified cDNA. Detection of mature miRNAs is achieved by in situ hybridization with locked nucleic acid probes. This part of the protocol involves a brief protease digestion, followed by an overnight hybridization, short low stringency wash and detection of the labeled probe. The key variables for this method include probe concentration and stringency conditions. Each miRNA in situ method takes 1 d. The final step of the protocol involves colabeling by immunohistochemistry for the putative target of the miRNA, which is done after the in situ hybridization step and takes a few hours. PMID- 19131964 TI - Sgt1, a co-chaperone of Hsp90 stabilizes Polo and is required for centrosome organization. AB - Sgt1 was described previously in yeast and humans to be a Hsp90 co-chaperone and required for kinetochore assembly. We have identified a mutant allele of Sgt1 in Drosophila and characterized its function. Mutations in sgt1 do not affect overall kinetochore assembly or spindle assembly checkpoint. sgt1 mutant cells enter less frequently into mitosis and arrest in a prometaphase-like state. Mutations in sgt1 severely compromise the organization and function of the mitotic apparatus. In these cells, centrioles replicate but centrosomes fail to mature, and pericentriolar material components do not localize normally resulting in highly abnormal spindles. Interestingly, a similar phenotype was described previously in Hsp90 mutant cells and correlated with a decrease in Polo protein levels. In sgt1 mutant neuroblasts, we also observe a decrease in overall levels of Polo. Overexpression of the kinase results in a substantial rescue of the centrosome defects; most cells form normal bipolar spindles and progress through mitosis normally. Taken together, these findings suggest that Sgt1 is involved in the stabilization of Polo allowing normal centrosome maturation, entry and progression though mitosis. PMID- 19131965 TI - The ubiquitin-editing enzyme A20 requires RNF11 to downregulate NF-kappaB signalling. AB - The RING domain protein RNF11 is overexpressed in breast cancers and promotes tumour growth factor-beta (TGF-beta) signalling. RNF11 has been proposed to regulate TGF-beta signalling by interacting with HECT- and SCF-type E3 ligases; however, the role of RNF11 in other signalling pathways is poorly understood. Here, we demonstrate a novel function of RNF11 as a negative regulator of NF kappaB and jun N-terminal kinase (JNK) signalling pathways. Knockdown of RNF11 with siRNA resulted in persistent tumour necrosis factor (TNF)- and lipopolysaccharide (LPS)-mediated NF-kappaB and JNK signalling. RNF11 interacted with the NF-kappaB inhibitor A20 and its regulatory protein TAX1BP1 in a stimulus dependent manner. RNF11 negatively regulated RIP1 and TRAF6 ubiquitination upon stimulation with TNF and LPS, respectively. Furthermore, RNF11 was required for A20 to interact with and inactivate RIP1 to inhibit TNF-mediated NF-kappaB activation. Our studies reveal that RNF11, together with TAX1BP1 and Itch, is an essential component of an A20 ubiquitin-editing protein complex that ensures transient activation of inflammatory signalling pathways. PMID- 19131966 TI - Conversion of the 2 Cl(-)/1 H+ antiporter ClC-5 in a NO3(-)/H+ antiporter by a single point mutation. AB - Several members of the CLC family are secondary active anion/proton exchangers, and not passive chloride channels. Among the exchangers, the endosomal ClC-5 protein that is mutated in Dent's disease shows an extreme outward rectification that precludes a precise determination of its transport stoichiometry from measurements of the reversal potential. We developed a novel imaging method to determine the absolute proton flux in Xenopus oocytes from the extracellular proton gradient. We determined a transport stoichiometry of 2 Cl(-)/1 H+. Nitrate uncoupled proton transport but mutating the highly conserved serine 168 to proline, as found in the plant NO3(-)/H+ antiporter atClCa, led to coupled NO3( )/H+ exchange. Among several amino acids tested at position 168, S168P was unique in mediating highly coupled NO3(-)/H+ exchange. We further found that ClC-5 is strongly stimulated by intracellular protons in an allosteric manner with an apparent pK of approximately 7.2. A 2:1 stoichiometry appears to be a general property of CLC anion/proton exchangers. Serine 168 has an important function in determining anionic specificity of the exchange mechanism. PMID- 19131967 TI - Sall2 is a novel p75NTR-interacting protein that links NGF signalling to cell cycle progression and neurite outgrowth. AB - By screening a fetal brain two-hybrid library with the death domain of the p75 neurotrophin receptor (NTR), we identified the Sall2 transcription factor as a novel interacting protein. Sall2 is a unique member of the Sall gene family, which is believed to be a tumour suppressor. Here, we show that Sall2 contains a p75NTR interaction domain not found in other Sall proteins and that p75NTR/Sall2 complexes co-immunoprecipitate from brain lysates. NGF dissociates p75NTR/Sall2 complexes and activates TrkA, which has an obligate function in the nuclear translocation of Sall2. NGF also increases Sall2 expression and this is mediated by p75NTR, but may not require TrkA. Depletion of Sall2 from cells decreases the expression and activity of p21(WAF1/CIP1), as well as the ability of NGF to induce growth arrest and the development of neurites. Overexpression of Sall2 activates p21(WAF1/CIP1), induces growth arrest, and promotes neurite outgrowth independently of NGF. These data establish Sall2 as a link between NTRs and transcriptional events that regulate the growth and development of neuronal cells. PMID- 19131968 TI - Repression of C. elegans microRNA targets at the initiation level of translation requires GW182 proteins. AB - MicroRNAs (miRNAs) repress target genes through a poorly defined antisense mechanism. Cell-free and cell-based assays have supported the idea that miRNAs repress their target mRNAs by blocking initiation of translation, whereas studies in animal models argued against this possibility. We examined endogenous targets of the let-7 miRNA, an important regulator of stem cell fates. We report that let 7 represses translation initiation in Caenorhabditis elegans, demonstrating this mode of action for the first time in an organism. Unexpectedly, although the lin 4 miRNA was previously reported to repress its targets at a step downstream of translation initiation, we also observe repression of translation initiation for this miRNA. This repressive mechanism, which frequently but not always coincides with transcript degradation, requires the GW182 proteins AIN-1 and AIN-2, and acts on several mRNAs targeted by different miRNAs. Our analysis of an expanded set of endogenous miRNA targets therefore indicates widespread repression of translation initiation under physiological conditions and establishes C. elegans as a genetic system for dissection of the underlying mechanisms. PMID- 19131969 TI - Remodelling of VipA/VipB tubules by ClpV-mediated threading is crucial for type VI protein secretion. AB - The recently identified type VI secretion systems (T6SS) have a crucial function in the virulence of various proteobacteria, including the human pathogen Vibrio cholerae. T6SS are encoded by a conserved gene cluster comprising approximately 15 open reading frames, mediating the appearance of Hcp and VgrG proteins in cell culture supernatants. Here, we analysed the function of the V. cholerae T6SS member ClpV, a specialized AAA+ protein. ClpV is crucial for a functional T6SS and interacts through its N-terminal domain with the VipA/VipB complex that is composed of two conserved and essential members of T6SS. Transferring ClpV substrate specificity to a distinct AAA+ protein involved in proteolysis caused degradation of VipA but not Hcp or VgrG2, suggesting that VipA rather than Hcp/VgrG2 functions as a primary ClpV substrate. Strikingly, VipA/VipB form tubular, cogwheel-like structures that are converted by a threading activity of ClpV into small complexes. ClpV-mediated remodelling of VipA/VipB tubules represents a crucial step in T6S, illuminating an unexpected role of an ATPase component in protein secretion. PMID- 19131970 TI - Ctk1 promotes dissociation of basal transcription factors from elongating RNA polymerase II. AB - As RNA polymerase II (RNApII) transitions from initiation to elongation, Mediator and the basal transcription factors TFIID, TFIIA, TFIIH, and TFIIE remain at the promoter as part of a scaffold complex, whereas TFIIB and TFIIF dissociate. The yeast Ctk1 kinase associates with elongation complexes and phosphorylates serine 2 in the YSPTSPS repeats of the Rpb1 C-terminal domain, a modification that couples transcription to mRNA 3'-end processing. The higher eukaryotic kinase Cdk9 not only performs a similar function, but also functions at the 5'-end of genes in the transition from initiation to elongation. In strains lacking Ctk1, many basal transcription factors cross-link throughout transcribed regions, apparently remaining associated with RNApII until it terminates. Consistent with this observation, preinitiation complexes formed on immobilized templates with transcription extracts lacking Ctk1 leave lower levels of the scaffold complex behind after escape. Taken together, these results suggest that Ctk1 is necessary for the release of RNApII from basal transcription factors. Interestingly, this function of Ctk1 is independent of its kinase activity, suggesting a structural function of the protein. PMID- 19131971 TI - The Axin1 scaffold protein promotes formation of a degradation complex for c-Myc. AB - Expression of the c-Myc proto-oncoprotein is tightly regulated in normal cells. Phosphorylation at two conserved residues, threonine58 (T58) and serine62 (S62), regulates c-Myc protein stability. In cancer cells, c-Myc can become aberrantly stabilized associated with altered T58 and S62 phosphorylation. A complex signalling cascade involving GSK3beta kinase, the Pin1 prolyl isomerase, and the PP2A-B56alpha phosphatase controls phosphorylation at these sites. We report here a novel role for the tumour suppressor scaffold protein Axin1 in facilitating the formation of a degradation complex for c-Myc containing GSK3beta, Pin1, and PP2A B56alpha. Although knockdown of Axin1 decreases the association of c-Myc with these proteins, reduces T58 and enhances S62 phosphorylation, and increases c-Myc stability, acute expression of Axin1 reduces c-Myc levels and suppresses c-Myc transcriptional activity. Moreover, the regulation of c-Myc by Axin1 is impaired in several tested cancer cell lines with known stabilization of c-Myc or loss of Axin1. This study provides critical insight into the regulation of c-Myc expression, how this can be disrupted in three cancer types, and adds to our knowledge of the tumour suppressor activity of Axin1. PMID- 19131973 TI - Begging bowls. PMID- 19131972 TI - Crystal structure of plant light-harvesting complex shows the active, energy transmitting state. AB - Plants dissipate excess excitation energy as heat by non-photochemical quenching (NPQ). NPQ has been thought to resemble in vitro aggregation quenching of the major antenna complex, light harvesting complex of photosystem II (LHC-II). Both processes are widely believed to involve a conformational change that creates a quenching centre of two neighbouring pigments within the complex. Using recombinant LHC-II lacking the pigments implicated in quenching, we show that they have no particular role. Single crystals of LHC-II emit strong, orientation dependent fluorescence with an emission maximum at 680 nm. The average lifetime of the main 680 nm crystal emission at 100 K is 1.31 ns, but only 0.39 ns for LHC II aggregates under identical conditions. The strong emission and comparatively long fluorescence lifetimes of single LHC-II crystals indicate that the complex is unquenched, and that therefore the crystal structure shows the active, energy transmitting state of LHC-II. We conclude that quenching of excitation energy in the light-harvesting antenna is due to the molecular interaction with external pigments in vitro or other pigment-protein complexes such as PsbS in vivo, and does not require a conformational change within the complex. PMID- 19131974 TI - Biotech sector ponders potential 'bloodbath'. PMID- 19131976 TI - Buzz around Campath proof-of-concept trial in MS. PMID- 19131979 TI - Doubts surround link between Bt cotton failure and farmer suicide. PMID- 19131983 TI - JAMA study casts cloud over biologic safety. PMID- 19131984 TI - Mark Roth. PMID- 19131985 TI - Compensation continues rise. PMID- 19131986 TI - Biotech's green gold? PMID- 19131987 TI - Industry and academia must avoid mismatching disclosures. PMID- 19131988 TI - Stem cell transplants: the power of peer-to-peer. PMID- 19131989 TI - Is FAO selling biotech short on biofuels? PMID- 19131991 TI - PRIME importance of pathology expertise. PMID- 19131992 TI - Quality by design for biopharmaceuticals. PMID- 19131994 TI - Patents versus patenting: implications of intellectual property protection for biological research. AB - A new survey shows scientists consider the proliferation of intellectual property protection to have a strongly negative effect on research. PMID- 19131997 TI - Looking ahead in cancer stem cell research. PMID- 19131996 TI - Crossing the rubicon. PMID- 19131998 TI - GPS navigation of the protein-stability landscape. PMID- 19132000 TI - Executive compensation at private life science companies in troubled economic times. PMID- 19132002 TI - Single-port surgery for donor nephrectomy: a new era in laparoscopic surgery? PMID- 19132003 TI - Photodynamic therapy for prostate cancer--a review of current status and future promise. AB - Debate is ongoing about the treatment of organ-confined prostate cancer, particularly in men who have low-risk disease detected by PSA screening. A balance is needed between the harms and benefits of treatment. New techniques are being developed that aim to offer similar treatment effects to current radical therapies, while reducing the associated harmful effects of these treatments. In this Review, we explore the potential of one such technique, photodynamic therapy (PDT), for the treatment of organ-confined prostate cancer. PDT uses a photosensitizing drug that is activated in the prostate by low-power laser light, delivered using optical fibers. The fibers are placed within needles in the prostate, guided by transrectal ultrasound and a perineal template. Following the activation of the photosensitizer by light, and the formation of reactive oxygen species, necrosis occurs at the site of interaction between the photosensitizer, light and oxygen. Clinical studies are underway to investigate the use of PDT for primary and salvage treatment of organ-confined prostate cancer. We review these studies, the potential strategies for enhanced photodynamic effects, and the current limitations of PDT for prostate cancer. PMID- 19132004 TI - Urologic complications of HIV and AIDS. AB - In recent years the nature of HIV infection has been dramatically transformed from an invariably fatal disease to a chronic disorder with a relatively benign course. Disease progression from HIV to AIDS and HIV-related mortality can be reduced effectively by several years of treatment with highly active antiretroviral therapy (HAART). For patients who do not have access to HAART, HIV infection continues to be a lethal disorder characterized by opportunistic infection with uncommon organisms (e.g. mycobacteria, fungi, parasites and viruses), as well as lethal malignancies such as Kaposi sarcoma, non-Hodgkin lymphoma and squamous cell carcinoma of the penis or cervix. In patients receiving HAART, urologic complications are likely to be caused by adverse effects of antiretroviral medication (e.g. indinavir urolithiasis) or disorders associated with aging, such as benign prostatic hyperplasia and prostate cancer. Prospective clinical trials have shown that adult male circumcision can reduce the rate of female to male HIV transmission by more than 50%; however, the development of preventive or curative modalities with 100% efficacy remains elusive. PMID- 19132005 TI - Subtypes of alpha1-adrenoceptors in BPH: future prospects for personalized medicine. AB - The alpha(1)-adrenoceptors (alpha(1)-ARs) are involved in regulation of prostatic smooth muscle tone, and are a critical mediator of lower urinary tract symptoms and pathophysiology in benign prostatic hyperplasia (BPH). As a result, alpha(1) AR antagonists are now used as first-line medical treatment for BPH. Three alpha(1)-AR subtypes (alpha(1a)-AR, alpha(1b)-AR, alpha(1d)-AR) have been identified on the basis of results of pharmacological and molecular cloning studies; however, the precise physiological role of individual alpha(1)-AR subtypes remains elusive. The expression levels of alpha(1)-AR subtypes in the prostate differ between patients, and individual differences in the genetic background of patients with BPH might be associated with variation in responses to subtype-selective alpha(1)-AR antagonists. In addition, single nucleotide polymorphism and microarray-based gene expression profiling studies might provide an opportunity to identify markers that predict clinical response and therapeutic tolerance to alpha(1)-AR antagonists. Further genomic studies will refine our knowledge of the functions of alpha(1)-AR subtypes, lead to new strategies for the clinical management of BPH and, perhaps, enable personalized treatment of BPH in the future. PMID- 19132006 TI - Ribosome hijacking: a role for small protein B during trans-translation. AB - Tight recognition of codon-anticodon pairings by the ribosome ensures the accuracy and fidelity of protein synthesis. In eubacteria, translational surveillance and ribosome rescue are performed by the 'tmRNA-SmpB' system (transfer messenger RNA-small protein B). Remarkably, entry and accommodation of aminoacylated-tmRNA into stalled ribosomes occur without a codon-anticodon interaction but in the presence of SmpB. Here, we show that within a stalled ribosome, SmpB interacts with the three universally conserved bases G530, A1492 and A1493 that form the 30S subunit decoding centre, in which canonical codon anticodon pairing occurs. The footprints at positions A1492 and A1493 of a small decoding centre, as well as on a set of conserved SmpB amino acids, were identified by nuclear magnetic resonance. Mutants at these residues display the same growth defects as for DeltasmpB strains. The SmpB protein has functional and structural similarities with initiation factor 1, and is proposed to be a functional mimic of the pairing between a codon and an anticodon. PMID- 19132007 TI - How does SIRT1 affect metabolism, senescence and cancer? AB - SIRT1 is a multifaceted, NAD(+)-dependent protein deacetylase that is involved in a wide variety of cellular processes from cancer to ageing. The function of SIRT1 in cancer is complex: SIRT1 has been shown to have oncogenic properties by downregulating p53 activity, but recent studies indicate that SIRT1 acts as a tumour suppressor in a mutated p53 background, raising intriguing questions regarding its mechanism of action. Here we discuss the current understanding of how SIRT1 functions in light of recent discoveries and propose that the net outcome of the seemingly opposite oncogenic and tumour-suppressive effects of SIRT1 depends on the status of p53. PMID- 19132008 TI - Do 'basal-like' breast cancers really exist? AB - It has been proposed that gene expression profiles will revolutionize the classification of breast cancer, eventually replacing histopathology with a more reproducible technology. These new approaches, combined with a better understanding of the cellular origins of breast cancer, should enable us to identify patient subgroups for more effective therapy. However, in such a rapidly advancing field it is essential that initial and thought-provoking results do not become established as 'facts' without question. This Opinion addresses some of the negatives and positives generated by the term 'basal-like' breast cancer, and questions its existence as an entity. PMID- 19132011 TI - On the abundance of EpCAM on cancer stem cells. PMID- 19132012 TI - Multisource feedback (MSF): supporting professional development. PMID- 19132009 TI - Senescence-messaging secretome: SMS-ing cellular stress. AB - Oncogene-induced cellular senescence constitutes a strong anti-proliferative response, which can be set in motion following either oncogene activation or loss of tumour suppressor signalling. It serves to limit the expansion of early neoplastic cells and as such is a potent cancer-protective response to oncogenic events. Recently emerging evidence points to a crucial role in oncogene-induced cellular senescence for the 'senescence-messaging secretome' or SMS, setting the stage for cross-talk between senescent cells and their environment. How are such signals integrated into a coordinated response and what are the implications of this unexpected finding? PMID- 19132013 TI - OTC Medications. PMID- 19132014 TI - Formocresol alternatives. PMID- 19132015 TI - Cautionary tale. PMID- 19132016 TI - Fully capable. PMID- 19132017 TI - Formocresol: a plea. PMID- 19132020 TI - High-flying dentistry. PMID- 19132022 TI - Honour awarded. PMID- 19132028 TI - An unusual complication of an inferior dental nerve block: a case report. AB - Local anaesthetic drugs are commonly used in dental practice, with few complications. We describe an unusual complication of an inferior dental nerve block where, as the needle was advanced through the mucosa, the patient experienced profound numbness and skin pallor in the distribution of the infra orbital nerve. We discuss the possible mechanism for this complication. PMID- 19132029 TI - Aviation dentistry: current concepts and practice. AB - BACKGROUND: With the growing number of air passengers, flight attendants, leisure pilots as well as military and airline pilots, dentists may increasingly encounter flight-related oral conditions requiring treatment. Moreover, dentists should prevent the creation of in-flight hazards when treating aircrew members. The aim of this article is to introduce the concepts of aviation (aerospace) medicine and dentistry. METHODS: Data were gathered to cover the following issues: head and facial barotraumas (barotrauma-related headache, external otitic barotrauma, barosinusitis and barotitis-media), dental barotrauma (barometric pressure-related tooth injury), barodontalgia (barometric pressure-related oro dental pain), and dental care for aircrews. RESULTS AND CONCLUSIONS: Special considerations have to be made when planning restorative, endodontic, prosthodontic and surgical treatment to an aircrew patient. This article supplies the dental practitioner with some diagnostic tools as well as treatment guidelines. Principles of prevention, periodic examination, dental-related flight restriction (grounding) and dental documentation (for forensic purposes) are described as well. PMID- 19132030 TI - Qat and its health effects. AB - In Southern Arabia and Eastern Africa, qat chewing is a widely practised socio cultural habit. It consists of placing the green-leaved plant into the mucobuccal fold and chewing it for several hours, with subsequent release of psychoactive agents. Qat chewing is often accompanied by smoking tobacco. The reported prevalence of qat chewing in Europe and North America is on the increase with global migration. Oral diseases reportedly associated with qat chewing include periodontitis, oral leukoplakia and oral cancer. However, precise data on the association of qat use with the development of oral cancer are sparse. The aim of this review is to 1) Educate health clinicians about qat usage and related oral/systemic health issues; and 2) Review the current literature regarding qat use and its association with oral disease but more specifically review its link with oral leukoplakia and oral squamous cell carcinoma (OSCC). To do this we searched the literature (PubMed, Science Direct and Scopus) to identify all relevant articles published over the last 20 years using a combination of terms 'qat', 'khat', 'kat', 'cathinone' and 'cathaedulis'. PMID- 19132037 TI - Educational innovations for dentistry. AB - Over the past four decades, teaching and learning in higher education has been informed by a variety of educational theories. The various initiatives that have been introduced as a consequence have all sought to improve students' educational experiences. However, such moves have not always been successful, as a series of three articles that the authors produced on dental chairside teaching have demonstrated. The first of these investigated the stakeholders' perceptions of this teaching at one dental school. The second evaluated chairside teaching on a UK wide scale, while the third provided educational tools to encourage collaboration among clinical educators to share good chairside teaching practices. What this paper seeks to do is to locate each of these themes within the wider context of a theoretical frame highlighting the need for dental education to be underpinned by such constructs. The aim of this paper is to help ensure that an informed learning experience is achieved for all students engaged in clinical dental education. PMID- 19132038 TI - Clinical audit and peer review scheme for the South West post-new 2006 dental contract: a report on progress so far. AB - With the introduction of personal dental services (PDS) into the South West the Local Assessment Panel (LAP) devised a new scheme consisting of 'cookbook' audits and piloted the scheme amongst the PDS dentists of South and West Devon, Somerset and Avon in 2005/2006. When the new contracting arrangements came into force, and in the absence of guidance from above, the LAP in consultation with the PCTs decided to consolidate the successful pilot audit scheme for PDS dentists and extend the new scheme to all the participating PCTs and their performers. The current scheme covers Devon, Somerset, Avon and Gloucester PCTs and is administrated by Mrs Jackie Derrick on behalf of Somerset PCT. All the audits showed improvement with the exception of the patient satisfaction survey where the first audit cycle showed an average patient satisfaction rating of 99% which cannot be improved on. We have redesigned this audit to try and make it more challenging and informative. The improvement in clinical record keeping was particularly marked. With the advent of new contractual arrangements in April 2009 it is essential that practitioners are able to demonstrate quality assurance in their practice and we believe that the South West scheme is a dentist friendly scheme, relevant to everyday dental practice. PMID- 19132054 TI - A cell-based screening to detect inhibitors of BRAF signaling pathway. PMID- 19132055 TI - Albidopyrone, a new alpha-pyrone-containing metabolite from marine-derived Streptomyces sp. NTK 227. AB - Albidopyrone, a new alpha-pyrone-containing secondary metabolite, was produced by Streptomyces sp. NTK 227, a strain isolated from Atlantic Ocean sediment and found to be a member of the Streptomyces albidoflavus 16S rRNA gene clade. The structure of the compound was determined by MS and NMR spectroscopy, and found to have a moderate inhibitory activity against protein-tyrosin phosphatase B. PMID- 19132056 TI - Ceramidastin, a novel bacterial ceramidase inhibitor, produced by Penicillium sp. Mer-f17067. AB - Decrease of ceramide in the skin is one of the aggravating factors of atopic dermatitis. The skin is often infected by ceramidase-producing bacteria, such as Pseudomonas aeruginosa. The bacterial ceramidase then degrades ceramide in the skin. To develop anti-atopic dermatitis drugs, we searched for ceramidase inhibitors, which led to the discovery of ceramidastin, a novel inhibitor of bacterial ceramidase, from the culture broth of Penicillium sp. Mer-f17067. Ceramidastin inhibited the bacterial ceramidase with an IC(50) value of 6.25 microg ml(-1). Here we describe the isolation, physicochemical properties, structure determination and biological activity of ceramidastin. PMID- 19132057 TI - Synergistic fungicidal activities of polymyxin B and ionophores, and their dependence on direct disruptive action of polymyxin B on fungal vacuole. AB - Polymyxin B (PMB) acts selectively on Gram-negative bacteria by electrostatic and hydrophobic interactions with anionic cell envelope components such as phospholipids and lipopolysaccharides. In this study, PMB was shown to exhibit marked fungicidal activity against yeasts and filamentous fungi in combination with ionophores such as salinomycin (SAM) and monensin (MON), which can selectively interact with monovalent cations. Ca(2+)-selective ionophores, A23187 and ionomycin, were absolutely ineffective in enhancing the fungicidal activity of PMB. SAM and MON increased the rate of cellular uptake of PMB possibly in favor of its intracellular action on the organelle. PMB could indeed directly disrupt the spherical membrane-enclosed architecture of the isolated vacuoles equally in the absence and presence of the ionophores. The loss of energy barrier for transmembrane transport of monovalent cations is considered to be a cause of enhanced incorporation of larger cationic compounds such as PMB across fungal plasma membrane. PMID- 19132058 TI - Micafungin: a sulfated echinocandin. AB - Micafungin is the second approved antifungal agent in the echinocandin series and is now used worldwide in chemotherapy for life-threatening fungal infections. It is water-soluble and is semi-synthesized from the acylated cyclic hexapeptide FR901379, a natural product from the fungus Coleophoma empetri F-11899, through enzymatic deacylation of FR901379, followed by chemical reacylation with the optimized N-acyl side chain. The water solubility of micafungin is ascribed to a sulfate moiety in the molecule. This feature differentiates micafungin from other echinocandin members. Micafungin is a potent inhibitor of 1,3-beta-glucan synthase, an enzyme necessary for cell-wall synthesis of several fungal pathogens. PMID- 19132059 TI - Chemical biology of natural indolocarbazole products: 30 years since the discovery of staurosporine. AB - Staurosporine was discovered at the Kitasato Institute in 1977 while screening for microbial alkaloids using chemical detection methods. It was during the same era that protein kinase C was discovered and oncogene v-src was shown to have protein kinase activity. Staurosporine was first isolated from a culture of Actinomyces that originated in a soil sample collected in Mizusawa City, Japan. Thereafter, indolocarbazole compounds have been isolated from a variety of organisms. The biosynthesis of staurosporine and related indolocarbazoles was finally elucidated during the past decade through genetic and biochemical studies. Subsequently, several novel indolocarbazoles have been produced using combinatorial biosynthesis. In 1986, 9 years since its discovery, staurosporine and related indolocarbazoles were shown to be nanomolar inhibitors of protein kinases. They can thus be viewed as forerunners of today's crop of novel anticancer drugs. The finding led many pharmaceutical companies to search for selective protein kinase inhibitors by screening natural products and through chemical synthesis. In the 1990s, imatinib, a Bcr-Abl tyrosine kinase inhibitor, was synthesized and, following human clinical trials for chronic myelogenous leukemia, it was approved for use in the USA in 2001. In 1992, mammalian topoisomerases were shown to be targets for indolocarbazoles. This opened up new possibilities in that indolocarbazole compounds could selectively interact with ATP-binding sites of not only protein kinases but also other proteins that had slight differences in ATP-binding sites. ABCG2, an ATP-binding cassette transporter, was recently identified as an important new target for indolocarbazoles. PMID- 19132060 TI - Genetic analysis of Helicobacter pylori clinical isolates suggests resistance to metronidazole can occur without the loss of functional rdxA. AB - Resistance to metronidazole (MTZ) in Helicobacter pylori is associated with mutations in rdxA, encoding an oxygen-insensitive NADPH nitroreductase, and mutations in frxA, encoding a NAD(P)H-flavin oxidoreductase. Despite this association, the strict correlation of MTZ resistance with mutations in rdxA or frxA is still controversial. In this study, rdxA allelic replacement was used to distinguish resistance-associated nucleotide mutations from the natural genetic diversity of H. pylori. Replacement with truncated rdxA resulted in MTZ resistance, whereas replacement with missense-mutated rdxA from resistant clinical isolates failed to yield MTZ resistance. Thus, although truncation of rdxA confers MTZ resistance in G27 H. pylori, MTZ resistance found in other clinical isolates is not due to the identified amino-acid substitutions. Three of our MTZ-resistant clinical isolates expressed functional rdxA and two of these also encoded full-length frxA. Therefore, MTZ resistance can arise in H. pylori possessing functional rdxA, suggesting that other factors are involved in MTZ resistance. PMID- 19132062 TI - Microbial drug discovery: 80 years of progress. AB - Microbes have made a phenomenal contribution to the health and well-being of people throughout the world. In addition to producing many primary metabolites, such as amino acids, vitamins and nucleotides, they are capable of making secondary metabolites, which constitute half of the pharmaceuticals on the market today and provide agriculture with many essential products. This review centers on these beneficial secondary metabolites, the discovery of which goes back 80 years to the time when penicillin was discovered by Alexander Fleming. PMID- 19132061 TI - Modification of the antibiotic olivomycin I at the 2'-keto group of the side chain. Novel derivatives, antitumor and topoisomerase I-poisoning activity. AB - A novel way of chemical modification of the antibiotic olivomycin I at the 2' keto group of the side chain of the aglycone moiety was developed. Reaction of olivomycin I with the carboxymethoxylamine hemihydrochloride gave the key intermediate, 2'-carboxymethoxime-olivomycin I, which was further reacted with different amines in the presence of benzotriazol-1-yl-oxy trispyrrolidinophosphonium hexafluorophosphate to give the corresponding amides. The antiproliferative and topoisomerase I (Topo-I)-poisoning activities of the novel derivatives were examined. One of the novel derivatives showed a marked inhibitory activity against Topo-I, a pronounced antitumor activity in in vivo experiments on mice bearing leukemia P-388 and lower toxic side effects compared with the parent olivomycin I. PMID- 19132063 TI - Discovery of okilactomycin and congeners from Streptomyces scabrisporus by antisense differential sensitivity assay targeting ribosomal protein S4. AB - Protein synthesis inhibition is a highly successful target for developing clinically effective and safe antibiotics. There are several targets within the ribosomal machinery, and small ribosomal protein S4 (RPSD) is one of the newer targets. Screening of microbial extracts using antisense-sensitized rpsD Staphylococcus aureus strain led to isolation of okilactomycin and four new congeners from Streptomyces scabrisporus. The major compound, okilactomycin, was the most active, with a minimum detection concentration of 3-12 microg ml(-1) against antisense assay, and showed an MIC of 4-16 microg ml(-1) against Gram positive bacteria, including S. aureus. The congeners were significantly less active in all assays, and all compounds showed a slight preferential inhibition of RNA synthesis over DNA and protein synthesis. Antisense technology, due to increased sensitivity, continues to yield new, even though weakly active, antibiotics. PMID- 19132064 TI - Expression of two human acyl-CoA:diacylglycerol acyltransferase isozymes in yeast and selectivity of microbial inhibitors toward the isozymes. AB - Two isozymes for human acyl-coenzyme A:diacylglycerol acyltransferase (DGAT), DGAT1 and DGAT2, were independently expressed in DGAT-deficient Saccharomyces cerevisiae to establish DGAT1- and DGAT2-S. cerevisiae. The selectivity of DGAT inhibitors of natural origin towards the isozymes was assessed in enzyme assays using the microsomal fractions prepared from DGAT1- and DGAT2-S. cerevisiae. Amidepsines and xanthohumol inhibited DGAT1 and DGAT2 with similar potency, whereas roselipins were found to inhibit DGAT2 selectively. PMID- 19132065 TI - Expression and function of the coxsackie and adenovirus receptor in Barrett's esophagus and associated neoplasia. AB - Cell surface presence of the coxsackie and adenovirus receptor (CAR) is considered a crucial prerequisite for the uptake of attenuated adenovirus. In cancers, however, a frequent loss of CAR has been noted potentially hampering the success of adenovirus-based therapy. In esophageal Barrett's carcinomas and its precursor lesions CAR presence has not been systematically determined yet. Immunohistochemical assessment in tissue specimens of 111 patients revealed CAR positivity in all cases of Barrett's esophagus, including various degrees of intraepithelial neoplasia. In contrast, no considerable CAR presence was seen in squamous esophageal epithelium. Among Barrett's carcinomas, 93% displayed CAR presence, whereas CAR-negativity was observed preferentially in advanced cancers. Aiming to evaluate whether this loss of CAR impacts tumor-biologic properties of esophageal adenocarcinomas we studied cell lines OE19 and OE33 and observed an increased proliferation, migration and invasion upon siRNA-mediated functional CAR knock down. In conclusion, our results indicate that CAR may provide a valuable target for adenovirus-based therapy of Barrett's carcinomas and its precursor lesions. These data do also suggest that CAR does not contribute substantially to carcinogenesis in Barrett's esophagus, however, it may be speculated that loss of CAR promotes tumor progression in advanced stages of Barrett's carcinomas. PMID- 19132066 TI - cGMP-dependent protein kinase: linking foraging to energy homeostasis. AB - Successful foraging is necessary for procurement of nutritional resources essential for an animal's survival. Maintenance of foraging and food acquisition is dependent on the ability to balance food intake and energy expenditure. This review examines the role of cGMP-dependent protein kinase (PKG) as a regulator of foraging behaviour, food acquisition, and energy balance. The role of PKG in food related behaviours is highly conserved among worms, flies, bees, ants, and mammals. A growing body of literature suggests that PKG plays an integral role in the component behaviours and physiologies underlying foraging behaviour. These include energy acquisition, nutrient absorption, nutrient allocation, nutrient storage, and energy use. New evidence suggests that PKG mediates both neural and physiological mechanisms underlying these processes. This review illustrates how investigating the role of PKG in energy homeostasis in a diversity of organisms can offer a broad perspective on the mechanisms mediating energy balance. PMID- 19132067 TI - (Epi)genomics and neurodevelopment in schizophrenia: monozygotic twins discordant for schizophrenia augment the search for disease-related (epi)genomic alterations. AB - Unlike stunning breakthroughs in the identification of genes for Mendelian disorders during the last three decades, gene identification in most complex disorders has been full of twists and turns and little progress. Doing more of the same will not guarantee success. The lessons learned argue for a need to reconsider genetic models that are appropriate for the disorder in question along with an interdisciplinary, systematic approach using genomic methods that have now become possible. We will use schizophrenia as an example to review the genetic progress to date that has been disappointing. We will argue that the causation of this complex disease may involve heterogeneous genomic changes of major effect. We will provide three approaches, retroviral transpositions, methylation, and copy number variations, to test this hypothesis. We will present arguments to suggest that such experiments will be most effective if undertaken on monozygotic twins. It will include our experience with associated experiments on the monozygotic twins discordant for schizophrenia. The results support that (epi)genomic changes of major effect, rather than accumulation of mutations of small effect, underlie the causation of this complex disease. More important, this experimental strategy will be an effective strategy for studies on other complex (behavioural) disorders as well. PMID- 19132068 TI - Mitotic and polytene chromosome analysis in the Mexican fruit fly, Anastrepha ludens (Loew) (Diptera: Tephritidae). AB - The present study constitutes the first attempt to construct a polytene chromosome map of an Anastrepha species, Anastrepha ludens (Loew), a major agricultural pest. The mitotic karyotype has a diploid complement of 12 acrocentric chromosomes, including five pairs of autosomes and an XX/XY sex chromosome pair. The analysis of salivary gland polytene chromosomes has shown a total number of five polytene elements that correspond to the five autosomes. The characteristic features and the most prominent landmarks of each chromosome are described. By comparing chromosome banding patterns, the possible chromosomal homology between A. ludens and Ceratitis capitata (Wiedemann) is presented. This work shows that polytene maps of A. ludens are suitable for cytogenetic studies in this species and may be used as reference for other Anastrepha species, most of which are also serious agricultural pests. PMID- 19132069 TI - Insect mitochondrial genomics 3: the complete mitochondrial genome sequences of representatives from two neuropteroid orders: a dobsonfly (order Megaloptera) and a giant lacewing and an owlfly (order Neuroptera). AB - We describe the complete mitochondrial genomes from representatives of two orders of the Neuropterida: a dobsonfly, Corydalus cornutus (Megaloptera: Corydalidae, GenBank Accession No. FJ171323), a giant lacewing Polystoechotes punctatus (Neuroptera: Polystoechotidae, FJ171325), and an owlfly, Ascaloptynx appendiculatus (Neuroptera: Ascalaphidae, FJ171324). The dobsonfly sequence is 15,687 base pairs with a major noncoding (A+T rich) region of approximately 967 bp. The gene content and organization of the dobsonfly is identical to that of most insects. The giant lacewing sequence is 16 036 bp with a major noncoding region of about 1123 bp, while the owlfly sequence is 15,877 bp with a major noncoding region of about 1066 bp. The two Neuroptera sequences include a transposition of two tRNA genes, tRNATrp and tRNACys. These tRNA genes are coded on opposite strands and overlap by seven residues in the standard insect mitochondrial gene arrangement. Thus, the transposition required a duplication of at least the region of overlap. It is likely that the transposition occurred by a duplication of both genes followed by deletion of one copy of each gene. Examination of this region in two other neuropteroid species, a snakefly, Agulla sp. (Raphidioptera: Raphidiidae), and an antlion, Myrmeleon immaculatus (Neuroptera: Myrmeleontidae), shows that the rearrangement is widespread in the order Neuroptera but not present in either of the other two orders of Neuropterida. PMID- 19132070 TI - Robustness of QTLs across germplasm pools using a model quantitative trait. AB - Knowledge of the inheritance of C-glycosyl flavone synthesis in maize (Zea mays L.) silk tissues has been acquired through detailed genetic studies involving primarily germplasm from the Corn Belt Dent race. To test the robustness of this genetic knowledge, we examined C-glycosyl flavone synthesis in a genetically distinct germplasm pool, popcorn. C-glycosyl flavone profiles and levels and the involvement of three specific genes/quantitative trait loci (p, pr1, and sm1) in C-glycosyl flavone synthesis were examined in popcorn germplasm representing at least two races and various diverse geographic regions. Twenty-four inbred lines and 23 hybrids involving these inbred lines and inbred line R17 were characterized for their flavone profiles and levels in silk tissues. Two F2 mapping populations were constructed to examine the involvement of p, pr1, and sm1 on C-glycosyl flavone synthesis. C-glycosyl flavone levels threefold higher than previously reported in Corn Dent Belt materials and a novel class of compounds were discovered. The gene action of sm1 was different, the functional p allele was not always dominant, and pr1 did not affect mays in synthesis. Based on this rather simplistic "model" quantitative trait, it appears that caution should be exercised when attempting to apply quantitative trait locus knowledge accumulated in one germplasm base to a germplasm base that is known to be distinctly unique. PMID- 19132071 TI - An X chromosome effect responsible for asymmetric reproductive isolation between male Drosophila virilis and heterospecific females. AB - Reproductive isolation between closely related species is expressed through uncoordinated courtship, failed fertilization, and (or) postzygotic barriers. Behavioural components of mating often form an initial barrier to hybridization between species. In many animals, females are responsible for mating discrimination in both intra- and interspecific crosses; males of Drosophila virilis group represent an exception to this trend. Using overall productivity tests, we show that a lower proportion of D. virilis males sire progeny when paired with a heterospecific female (Drosophila novamexicana or Drosophila americana texana) for 2 weeks. This suggests male mate discrimination or some other kind of asymmetrical incompatibility in courtship and mating or early zygote mortality. We used males from D. virilis-D. novamexicana and from D. virilis-D. a. texana backcross populations to map chromosome effects responsible for male reproductive isolation. Results from the analysis of both backcross male populations indicate a major X chromosome effect. Further, we conduct a male behavioural analysis to show that D. virilis males significantly fail to continue courtship after the first step of courtship, when they tap heterospecific females. The combined results of a major X chromosome effect and the observation that D. virilis males walk away from females after tapping suggest that future studies should concentrate on the identification of X-linked genes affecting the ability of males to recognize conspecific females. PMID- 19132072 TI - Repetitive genome elements in a European corn borer, Ostrinia nubilalis, bacterial artificial chromosome library were indicated by bacterial artificial chromosome end sequencing and development of sequence tag site markers: implications for lepidopteran genomic research. AB - The European corn borer, Ostrinia nubilalis, is a serious pest of food, fiber, and biofuel crops in Europe, North America, and Asia and a model system for insect olfaction and speciation. A bacterial artificial chromosome library constructed for O. nubilalis contains 36 864 clones with an estimated average insert size of >or=120 kb and genome coverage of 8.8-fold. Screening OnB1 clones comprising approximately 2.76 genome equivalents determined the physical position of 24 sequence tag site markers, including markers linked to ecologically important and Bacillus thuringiensis toxin resistance traits. OnB1 bacterial artificial chromosome end sequence reads (GenBank dbGSS accessions ET217010 to ET217273) showed homology to annotated genes or expressed sequence tags and identified repetitive genome elements, O. nubilalis miniature subterminal inverted repeat transposable elements (OnMITE01 and OnMITE02), and ezi-like long interspersed nuclear elements. Mobility of OnMITE01 was demonstrated by the presence or absence in O. nubilalis of introns at two different loci. A (GTCT)n tetranucleotide repeat at the 5' ends of OnMITE01 and OnMITE02 are evidence for transposon-mediated movement of lepidopteran microsatellite loci. The number of repetitive elements in lepidopteran genomes will affect genome assembly and marker development. Single-locus sequence tag site markers described here have downstream application for integration within linkage maps and comparative genomic studies. PMID- 19132073 TI - Isolation of expressed sequences from a specific chromosome of Thinopyrum intermedium infected by BYDV. AB - To map important ESTs to specific chromosomes and (or) chromosomal regions is difficult in hexaploid wheat because of its large genome size and serious interference of homoeologous sequences. Large-scale EST sequencing and subsequent chromosome localization are both laborious and time-consuming. The wheat alien addition line TAi-27 contains a pair of chromosomes of Thinopyrum intermedium (Host) Barkworth & D.R. Dewey that carry the resistance gene against barley yellow dwarf virus. In this research, we developed a modified technique based on chromosome microdissection and hybridization-specific amplification to isolate expressed sequences from the alien chromosome of TAi-27 by hybridization between the DNA of the microdissected alien chromosome and cDNA of Th. intermedium infected by barley yellow dwarf virus. Twelve clones were selected, sequenced, and analyzed. Three of them were unknown genes without any hit in the GenBank database and the other nine were highly homologous with ESTs of wheat, barley, and (or) other plants in Gramineae induced by abiotic or biotic stress. The method used in this research to isolate expressed sequences from a specific chromosome has the following advantages: (i) the obtained expressed sequences are larger in size and have 3' end information and (ii) the operation is less complicated. It would be an efficient improved method for genomics and functional genomics research of polyploid plants, especially for EST development and mapping. The obtained expressed sequence data are also informative in understanding the resistance genes on the alien chromosome of TAi-27. PMID- 19132074 TI - Identification and characterization of the WRKY transcription factor family in Pinus monticola. AB - The WRKY gene family represents an ancient and highly complex group of transcription factors involved in signal transduction pathways of numerous plant developmental processes and host defense response. Up to now, most WRKY proteins have been identified in a few angiosperm species. Identification of WRKY genes in a conifer species would facilitate a comprehensive understanding of the evolutionary and function-adaptive process of this superfamily in plants. We performed PCR on genomic DNA to clone WRKY sequences from western white pine (Pinus monticola), one of the most valuable conifer species endangered by white pine blister rust (Cronartium ribicola). In total, 83 P. monticola WRKY (PmWRKY) sequences were identified using degenerate primers targeted to the WRKY domain. A phylogenetic analysis revealed that PmWRKY members fell into four major groups (1, 2a+2b, 2c, and 2d+2e) described in Arabidopsis and rice. Because of high genetic diversity of the PmWRKY family, a modified AFLP method was used to detect DNA polymorphism of this gene family. Polymorphic fragments accounted for 17%-35% of total PCR products in the AFLP profiles. Among them, one WRKY AFLP marker was linked to the major resistance gene (Cr2) against C. ribicola. The results of this study provide basic genomic information for a conifer WRKY gene family, which will pave the way for elucidating gene evolutionary mechanisms in plants and unveiling the precise roles of PmWRKY in conifer development and defense response. PMID- 19132075 TI - A quantitative genomic imbalance gene expression assay in a hexaploid species: wheat (Triticum aestivum). AB - Responses to allopolyploidy include unequal expression of duplicated genes, gene silencing, and sometimes genomic rearrangements. In plants, the relationship between allelic expression differences arising from changes in regulatory regions and the resulting phenotype is poorly understood because of the complexity of their genomes and lack of efficient methodology to identify regulatory variation. Identifying functionally important regulatory variation in crops such as hexaploid wheat (Triticum aestivum) is in its infancy. More knowledge is required about the contribution of participatory genomes to its transcriptome. In this paper, we demonstrate the use of allelic imbalance assays to quantify relative expression levels across tissues and growth regimes of homoeologous transcripts of the A, B, and D genomes. Polymorphisms in the type I thionins have been used as an example. We show that expression levels vary markedly and interactively over all factors. For this gene, the B genome is the smallest contributor to the transcriptome and the D genome the largest. As additional sequence information is accumulated across genomes, this assay will allow the simple study of relative expression across multiple homeologous loci. PMID- 19132076 TI - A molecular marker closely linked to the region of Rht-D1c and Ms2 genes in common wheat (Triticum aestivum). AB - A male sterile wheat mutant, Triticum aestivum L. 'Taigu', was found in a wheat field in China in 1972. The male sterility was controlled by a single dominant gene that was referred to as Ms2. Recently, this gene was found to be linked to a dwarfing gene through crossing Taigu with the short wheat T. aestivum 'Ai-Bian 1' carrying the dwarfing gene Rht-D1c. The objective of this study was to develop molecular markers linked to the male sterility Ms2 gene in common wheat. One hundred and twenty-two near-isogenic lines were developed through backcrossing and sib intercrossing and used as the mapping population for the development of molecular markers. Bulked segregant analysis was used to screen 48 pairs of SSR primers, and a marker, MS2-WMC617, was identified closely linked to the male sterile Ms2 gene that mapped at the distal position of chromosome arm 4DS. The use of the molecular marker MS2-WMC617 can facilitate recurrent selection in a wheat breeding program based on marker-assisted selection. PMID- 19132077 TI - Allopolyploidization reduces alternative splicing efficiency for transcripts of the wheat DREB2 homolog, WDREB2. AB - The wheat DREB2 homolog, WDREB2, encodes an EREBP/AP2-type transcription factor that activates downstream Cor/Lea genes and plays important roles in abiotic stress responses. WDREB2 expression is posttranscriptionally regulated by alternative splicing. In this study, the alternative splicing patterns of WDREB2 transcripts were compared among wheats of different ploidy levels. In diploid progenitors, alternative splicing rapidly produced functional forms from the WDREB2 transcripts and the levels of the nonfunctional form gradually decreased in response to drought stress; however, in hexaploid wheat lines, including both cultivars and synthetic lines, the nonfunctional form failed to decrease. The accumulation levels of the three spliced forms in synthetic allopolyploids were not equal to the sum of those in the parental lines. These results indicated that allopolyploidization during wheat polyploid evolution inhibited efficient alternative splicing of WDREB2 transcripts. PMID- 19132078 TI - Deterministic mechanical model of T-killer cell polarization reproduces the wandering of aim between simultaneously engaged targets. AB - T-killer cells of the immune system eliminate virus-infected and tumorous cells through direct cell-cell interactions. Reorientation of the killing apparatus inside the T cell to the T-cell interface with the target cell ensures specificity of the immune response. The killing apparatus can also oscillate next to the cell-cell interface. When two target cells are engaged by the T cell simultaneously, the killing apparatus can oscillate between the two interface areas. This oscillation is one of the most striking examples of cell movements that give the microscopist an unmechanistic impression of the cell's fidgety indecision. We have constructed a three-dimensional, numerical biomechanical model of the molecular-motor-driven microtubule cytoskeleton that positions the killing apparatus. The model demonstrates that the cortical pulling mechanism is indeed capable of orienting the killing apparatus into the functional position under a range of conditions. The model also predicts experimentally testable limitations of this commonly hypothesized mechanism of T-cell polarization. After the reorientation, the numerical solution exhibits complex, multidirectional, multiperiodic, and sustained oscillations in the absence of any external guidance or stochasticity. These computational results demonstrate that the strikingly animate wandering of aim in T-killer cells has a purely mechanical and deterministic explanation. PMID- 19132080 TI - Evolutionary sequence modeling for discovery of peptide hormones. AB - There are currently a large number of "orphan" G-protein-coupled receptors (GPCRs) whose endogenous ligands (peptide hormones) are unknown. Identification of these peptide hormones is a difficult and important problem. We describe a computational framework that models spatial structure along the genomic sequence simultaneously with the temporal evolutionary path structure across species and show how such models can be used to discover new functional molecules, in particular peptide hormones, via cross-genomic sequence comparisons. The computational framework incorporates a priori high-level knowledge of structural and evolutionary constraints into a hierarchical grammar of evolutionary probabilistic models. This computational method was used for identifying novel prohormones and the processed peptide sites by producing sequence alignments across many species at the functional-element level. Experimental results with an initial implementation of the algorithm were used to identify potential prohormones by comparing the human and non-human proteins in the Swiss-Prot database of known annotated proteins. In this proof of concept, we identified 45 out of 54 prohormones with only 44 false positives. The comparison of known and hypothetical human and mouse proteins resulted in the identification of a novel putative prohormone with at least four potential neuropeptides. Finally, in order to validate the computational methodology, we present the basic molecular biological characterization of the novel putative peptide hormone, including its identification and regional localization in the brain. This species comparison, HMM-based computational approach succeeded in identifying a previously undiscovered neuropeptide from whole genome protein sequences. This novel putative peptide hormone is found in discreet brain regions as well as other organs. The success of this approach will have a great impact on our understanding of GPCRs and associated pathways and help to identify new targets for drug development. PMID- 19132079 TI - An analysis of enzyme kinetics data for mitochondrial DNA strand termination by nucleoside reverse transcription inhibitors. AB - Nucleoside analogs used in antiretroviral treatment have been associated with mitochondrial toxicity. The polymerase-gamma hypothesis states that this toxicity stems from the analogs' inhibition of the mitochondrial DNA polymerase (polymerase-gamma) leading to mitochondrial DNA (mtDNA) depletion. We have constructed a computational model of the interaction of polymerase-gamma with activated nucleoside and nucleotide analog drugs, based on experimentally measured reaction rates and base excision rates, together with the mtDNA genome size, the human mtDNA sequence, and mitochondrial dNTP concentrations. The model predicts an approximately 1000-fold difference in the activated drug concentration required for a 50% probability of mtDNA strand termination between the activated di-deoxy analogs d4T, ddC, and ddI (activated to ddA) and the activated forms of the analogs 3TC, TDF, AZT, FTC, and ABC. These predictions are supported by experimental and clinical data showing significantly greater mtDNA depletion in cell culture and patient samples caused by the di-deoxy analog drugs. For zidovudine (AZT) we calculated a very low mtDNA replication termination probability, in contrast to its reported mitochondrial toxicity in vitro and clinically. Therefore AZT mitochondrial toxicity is likely due to a mechanism that does not involve strand termination of mtDNA replication. PMID- 19132081 TI - Why is the correlation between gene importance and gene evolutionary rate so weak? AB - One of the few commonly believed principles of molecular evolution is that functionally more important genes (or DNA sequences) evolve more slowly than less important ones. This principle is widely used by molecular biologists in daily practice. However, recent genomic analysis of a diverse array of organisms found only weak, negative correlations between the evolutionary rate of a gene and its functional importance, typically measured under a single benign lab condition. A frequently suggested cause of the above finding is that gene importance determined in the lab differs from that in an organism's natural environment. Here, we test this hypothesis in yeast using gene importance values experimentally determined in 418 lab conditions or computationally predicted for 10,000 nutritional conditions. In no single condition or combination of conditions did we find a much stronger negative correlation, which is explainable by our subsequent finding that always-essential (enzyme) genes do not evolve significantly more slowly than sometimes-essential or always-nonessential ones. Furthermore, we verified that functional density, approximated by the fraction of amino acid sites within protein domains, is uncorrelated with gene importance. Thus, neither the lab-nature mismatch nor a potentially biased among-gene distribution of functional density explains the observed weakness of the correlation between gene importance and evolutionary rate. We conclude that the weakness is factual, rather than artifactual. In addition to being weakened by population genetic reasons, the correlation is likely to have been further weakened by the presence of multiple nontrivial rate determinants that are independent from gene importance. These findings notwithstanding, we show that the principle of slower evolution of more important genes does have some predictive power when genes with vastly different evolutionary rates are compared, explaining why the principle can be practically useful despite the weakness of the correlation. PMID- 19132082 TI - Role of intraspecies recombination in the spread of pathogenicity islands within the Escherichia coli species. AB - Horizontal gene transfer is a key step in the evolution of bacterial pathogens. Besides phages and plasmids, pathogenicity islands (PAIs) are subjected to horizontal transfer. The transfer mechanisms of PAIs within a certain bacterial species or between different species are still not well understood. This study is focused on the High-Pathogenicity Island (HPI), which is a PAI widely spread among extraintestinal pathogenic Escherichia coli and serves as a model for horizontal transfer of PAIs in general. We applied a phylogenetic approach using multilocus sequence typing on HPI-positive and -negative natural E. coli isolates representative of the species diversity to infer the mechanism of horizontal HPI transfer within the E. coli species. In each strain, the partial nucleotide sequences of 6 HPI-encoded genes and 6 housekeeping genes of the genomic backbone, as well as DNA fragments immediately upstream and downstream of the HPI were compared. This revealed that the HPI is not solely vertically transmitted, but that recombination of large DNA fragments beyond the HPI plays a major role in the spread of the HPI within E. coli species. In support of the results of the phylogenetic analyses, we experimentally demonstrated that HPI can be transferred between different E. coli strains by F-plasmid mediated mobilization. Sequencing of the chromosomal DNA regions immediately upstream and downstream of the HPI in the recipient strain indicated that the HPI was transferred and integrated together with HPI-flanking DNA regions of the donor strain. The results of this study demonstrate for the first time that conjugative transfer and homologous DNA recombination play a major role in horizontal transfer of a pathogenicity island within the species E. coli. PMID- 19132083 TI - A novel co-crystal structure affords the design of gain-of-function lentiviral integrase mutants in the presence of modified PSIP1/LEDGF/p75. AB - Lens epithelium derived growth factor (LEDGF), also known as PC4 and SFRS1 interacting protein 1 (PSIP1) and transcriptional co-activator p75, is the cellular binding partner of lentiviral integrase (IN) proteins. LEDGF accounts for the characteristic propensity of Lentivirus to integrate within active transcription units and is required for efficient viral replication. We now present a crystal structure containing the N-terminal and catalytic core domains (NTD and CCD) of HIV-2 IN in complex with the IN binding domain (IBD) of LEDGF. The structure extends the known IN-LEDGF interface, elucidating primarily charge charge interactions between the NTD of IN and the IBD. A constellation of acidic residues on the NTD is characteristic of lentiviral INs, and mutations of the positively charged residues on the IBD severely affect interaction with all lentiviral INs tested. We show that the novel NTD-IBD contacts are critical for stimulation of concerted lentiviral DNA integration by LEDGF in vitro and for its function during the early steps of HIV-1 replication. Furthermore, the new structural details enabled us to engineer a mutant of HIV-1 IN that primarily functions only when presented with a complementary LEDGF mutant. These findings provide structural basis for the high affinity lentiviral IN-LEDGF interaction and pave the way for development of LEDGF-based targeting technologies for gene therapy. PMID- 19132084 TI - A role for the unfolded protein response (UPR) in virulence and antifungal susceptibility in Aspergillus fumigatus. AB - Filamentous fungi rely heavily on the secretory pathway, both for the delivery of cell wall components to the hyphal tip and the production and secretion of extracellular hydrolytic enzymes needed to support growth on polymeric substrates. Increased demand on the secretory system exerts stress on the endoplasmic reticulum (ER), which is countered by the activation of a coordinated stress response pathway termed the unfolded protein response (UPR). To determine the contribution of the UPR to the growth and virulence of the filamentous fungal pathogen Aspergillus fumigatus, we disrupted the hacA gene, encoding the major transcriptional regulator of the UPR. The DeltahacA mutant was unable to activate the UPR in response to ER stress and was hypersensitive to agents that disrupt ER homeostasis or the cell wall. Failure to induce the UPR did not affect radial growth on rich medium at 37 degrees C, but cell wall integrity was disrupted at 45 degrees C, resulting in a dramatic loss in viability. The DeltahacA mutant displayed a reduced capacity for protease secretion and was growth-impaired when challenged to assimilate nutrients from complex substrates. In addition, the DeltahacA mutant exhibited increased susceptibility to current antifungal agents that disrupt the membrane or cell wall and had attenuated virulence in multiple mouse models of invasive aspergillosis. These results demonstrate the importance of ER homeostasis to the growth and virulence of A. fumigatus and suggest that targeting the UPR, either alone or in combination with other antifungal drugs, would be an effective antifungal strategy. PMID- 19132085 TI - Arabidopsis CULLIN3 genes regulate primary root growth and patterning by ethylene dependent and -independent mechanisms. AB - CULLIN3 (CUL3) together with BTB-domain proteins form a class of Cullin-RING ubiquitin ligases (called CRL3s) that control the rapid and selective degradation of important regulatory proteins in all eukaryotes. Here, we report that in the model plant Arabidopsis thaliana, CUL3 regulates plant growth and development, not only during embryogenesis but also at post-embryonic stages. First, we show that CUL3 modulates the emission of ethylene, a gaseous plant hormone that is an important growth regulator. A CUL3 hypomorphic mutant accumulates ACS5, the rate limiting enzyme in ethylene biosynthesis and as a consequence exhibits a constitutive ethylene response. Second, we provide evidence that CUL3 regulates primary root growth by a novel ethylene-dependant pathway. In particular, we show that CUL3 knockdown inhibits primary root growth by reducing root meristem size and cell number. This phenotype is suppressed by ethylene-insensitive or resistant mutations. Finally, we identify a function of CUL3 in distal root patterning, by a mechanism that is independent of ethylene. Thus, our work highlights that CUL3 is essential for the normal division and organisation of the root stem cell niche and columella root cap cells. PMID- 19132086 TI - Control of stochastic gene expression by host factors at the HIV promoter. AB - The HIV promoter within the viral long terminal repeat (LTR) orchestrates many aspects of the viral life cycle, from the dynamics of viral gene expression and replication to the establishment of a latent state. In particular, after viral integration into the host genome, stochastic fluctuations in viral gene expression amplified by the Tat positive feedback loop can contribute to the formation of either a productive, transactivated state or an inactive state. In a significant fraction of cells harboring an integrated copy of the HIV-1 model provirus (LTR-GFP-IRES-Tat), this bimodal gene expression profile is dynamic, as cells spontaneously and continuously flip between active (Bright) and inactive (Off) expression modes. Furthermore, these switching dynamics may contribute to the establishment and maintenance of proviral latency, because after viral integration long delays in gene expression can occur before viral transactivation. The HIV-1 promoter contains cis-acting Sp1 and NF-kappaB elements that regulate gene expression via the recruitment of both activating and repressing complexes. We hypothesized that interplay in the recruitment of such positive and negative factors could modulate the stability of the Bright and Off modes and thereby alter the sensitivity of viral gene expression to stochastic fluctuations in the Tat feedback loop. Using model lentivirus variants with mutations introduced in the Sp1 and NF-kappaB elements, we employed flow cytometry, mRNA quantification, pharmacological perturbations, and chromatin immunoprecipitation to reveal significant functional differences in contributions of each site to viral gene regulation. Specifically, the Sp1 sites apparently stabilize both the Bright and the Off states, such that their mutation promotes noisy gene expression and reduction in the regulation of histone acetylation and deacetylation. Furthermore, the NF-kappaB sites exhibit distinct properties, with kappaB site I serving a stronger activating role than kappaB site II. Moreover, Sp1 site III plays a particularly important role in the recruitment of both p300 and RelA to the promoter. Finally, analysis of 362 clonal cell populations infected with the viral variants revealed that mutations in any of the Sp1 sites yield a 6-fold higher frequency of clonal bifurcation compared to that of the wild-type promoter. Thus, each Sp1 and NF-kappaB site differentially contributes to the regulation of viral gene expression, and Sp1 sites functionally "dampen" transcriptional noise and thereby modulate the frequency and maintenance of this model of viral latency. These results may have biomedical implications for the treatment of HIV latency. PMID- 19132088 TI - Evolution of regulatory sequences in 12 Drosophila species. AB - Characterization of the evolutionary constraints acting on cis-regulatory sequences is crucial to comparative genomics and provides key insights on the evolution of organismal diversity. We study the relationships among orthologous cis-regulatory modules (CRMs) in 12 Drosophila species, especially with respect to the evolution of transcription factor binding sites, and report statistical evidence in favor of key evolutionary hypotheses. Binding sites are found to have position-specific substitution rates. However, the selective forces at different positions of a site do not act independently, and the evidence suggests that constraints on sites are often based on their exact binding affinities. Binding site loss is seen to conform to a molecular clock hypothesis. The rate of site loss is transcription factor-specific and depends on the strength of binding and, in some cases, the presence of other binding sites in close proximity. Our analysis is based on a novel computational method for aligning orthologous CRMs on a tree, which rigorously accounts for alignment uncertainties and exploits binding site predictions through a unified probabilistic framework. Finally, we report weak purifying selection on short deletions, providing important clues about overall spatial constraints on CRMs. Our results present a complex picture of regulatory sequence evolution, with substantial plasticity that depends on a number of factors. The insights gained in this study will help us to understand the combinatorial control of gene regulation and how it evolves. They will pave the way for theoretical models that are cognizant of the important determinants of regulatory sequence evolution and will be critical in genome-wide identification of non-coding sequences under purifying or positive selection. PMID- 19132087 TI - A genome-wide association study identifies novel and functionally related susceptibility Loci for Kawasaki disease. AB - Kawasaki disease (KD) is a pediatric vasculitis that damages the coronary arteries in 25% of untreated and approximately 5% of treated children. Epidemiologic data suggest that KD is triggered by unidentified infection(s) in genetically susceptible children. To investigate genetic determinants of KD susceptibility, we performed a genome-wide association study (GWAS) in 119 Caucasian KD cases and 135 matched controls with stringent correction for possible admixture, followed by replication in an independent cohort and subsequent fine-mapping, for a total of 893 KD cases plus population and family controls. Significant associations of 40 SNPs and six haplotypes, identifying 31 genes, were replicated in an independent cohort of 583 predominantly Caucasian KD families, with NAALADL2 (rs17531088, p(combined) = 1.13 x 10(-6)) and ZFHX3 (rs7199343, p(combined) = 2.37 x 10(-6)) most significantly associated. Sixteen associated variants with a minor allele frequency of >0.05 that lay within or close to known genes were fine-mapped with HapMap tagging SNPs in 781 KD cases, including 590 from the discovery and replication stages. Original or tagging SNPs in eight of these genes replicated the original findings, with seven genes having further significant markers in adjacent regions. In four genes (ZFHX3, NAALADL2, PPP1R14C, and TCP1), the neighboring markers were more significantly associated than the originally associated variants. Investigation of functional relationships between the eight fine-mapped genes using Ingenuity Pathway Analysis identified a single functional network (p = 10(-13)) containing five fine-mapped genes-LNX1, CAMK2D, ZFHX3, CSMD1, and TCP1-with functional relationships potentially related to inflammation, apoptosis, and cardiovascular pathology. Pair-wise blood transcript levels were measured during acute and convalescent KD for all fine-mapped genes, revealing a consistent trend of significantly reduced transcript levels prior to treatment. This is one of the first GWAS in an infectious disease. We have identified novel, plausible, and functionally related variants associated with KD susceptibility that may also be relevant to other cardiovascular diseases. PMID- 19132090 TI - A cis-acting diversification activator both necessary and sufficient for AID mediated hypermutation. AB - Hypermutation of the immunoglobulin (Ig) genes requires Activation Induced cytidine Deaminase (AID) and transcription, but it remains unclear why other transcribed genes of B cells do not mutate. We describe a reporter transgene crippled by hypermutation when inserted into or near the Ig light chain (IgL) locus of the DT40 B cell line yet stably expressed when inserted into other chromosomal positions. Step-wise deletions of the IgL locus revealed that a sequence extending for 9.8 kilobases downstream of the IgL transcription start site confers the hypermutation activity. This sequence, named DIVAC for diversification activator, efficiently activates hypermutation when inserted at non-Ig loci. The results significantly extend previously reported findings on AID mediated gene diversification. They show by both deletion and insertion analyses that cis-acting sequences predispose neighboring transcription units to hypermutation. PMID- 19132089 TI - Genomic survey of the non-cultivatable opportunistic human pathogen, Enterocytozoon bieneusi. AB - Enterocytozoon bieneusi is the most common microsporidian associated with human disease, particularly in the immunocompromised population. In the setting of HIV infection, it is associated with diarrhea and wasting syndrome. Like all microsporidia, E. bieneusi is an obligate, intracellular parasite, but unlike others, it is in direct contact with the host cell cytoplasm. Studies of E. bieneusi have been greatly limited due to the absence of genomic data and lack of a robust cultivation system. Here, we present the first large-scale genomic dataset for E. bieneusi. Approximately 3.86 Mb of unique sequence was generated by paired end Sanger sequencing, representing about 64% of the estimated 6 Mb genome. A total of 3,804 genes were identified in E. bieneusi, of which 1,702 encode proteins with assigned functions. Of these, 653 are homologs of Encephalitozoon cuniculi proteins. Only one E. bieneusi protein with assigned function had no E. cuniculi homolog. The shared proteins were, in general, evenly distributed among the functional categories, with the exception of a dearth of genes encoding proteins associated with pathways for fatty acid and core carbon metabolism. Short intergenic regions, high gene density, and shortened protein coding sequences were observed in the E. bieneusi genome, all traits consistent with genomic compaction. Our findings suggest that E. bieneusi is a likely model for extreme genome reduction and host dependence. PMID- 19132091 TI - Ggamma1, a downstream target for the hmgcr-isoprenoid biosynthetic pathway, is required for releasing the Hedgehog ligand and directing germ cell migration. AB - The isoprenoid biosynthetic pathway leading from the production of mevalonate by HMGCoA reductase (Hmgcr) to the geranylation of the G protein subunit, Ggamma1, plays an important role in cardiac development in the fly. Hmgcr has also been implicated in the release of the signaling molecule Hedgehog (Hh) from hh expressing cells and in the production of an attractant that directs primordial germ cells to migrate to the somatic gonadal precursor cells (SGPs). The studies reported here indicate that this same hmgcr-->Ggamma1 pathway provides a novel post-translational mechanism for modulating the range and activity of the Hh signal produced by hh expressing cells. We show that, like hmgcr, ggamma1 and quemao (which encodes the enzyme, geranylgeranyl diphosphate synthetase, that produces the substrate for geranylation of Ggamma1) are components of the hh signaling pathway and are required for the efficient release of the Hh ligand from hh expressing cells. We also show that the hmgcr-->Ggamma1 pathway is linked to production of the germ cell attractant by the SGPs through its ability to enhance the potency of the Hh signal. We show that germ cell migration is disrupted by the loss or gain of ggamma1 activity, by trans-heterozygous combinations between ggamma1 and either hmgcr or hh mutations, and by ectopic expression of dominant negative Ggamma1 proteins that cannot be geranylated. PMID- 19132092 TI - Germ warfare in a microbial mat community: CRISPRs provide insights into the co evolution of host and viral genomes. AB - CRISPR arrays and associated cas genes are widespread in bacteria and archaea and confer acquired resistance to viruses. To examine viral immunity in the context of naturally evolving microbial populations we analyzed genomic data from two thermophilic Synechococcus isolates (Syn OS-A and Syn OS-B') as well as a prokaryotic metagenome and viral metagenome derived from microbial mats in hotsprings at Yellowstone National Park. Two distinct CRISPR types, distinguished by the repeat sequence, are found in both the Syn OS-A and Syn OS-B' genomes. The genome of Syn OS-A contains a third CRISPR type with a distinct repeat sequence, which is not found in Syn OS-B', but appears to be shared with other microorganisms that inhabit the mat. The CRISPR repeats identified in the microbial metagenome are highly conserved, while the spacer sequences (hereafter referred to as "viritopes" to emphasize their critical role in viral immunity) were mostly unique and had no high identity matches when searched against GenBank. Searching the viritopes against the viral metagenome, however, yielded several matches with high similarity some of which were within a gene identified as a likely viral lysozyme/lysin protein. Analysis of viral metagenome sequences corresponding to this lysozyme/lysin protein revealed several mutations all of which translate into silent or conservative mutations which are unlikely to affect protein function, but may help the virus evade the host CRISPR resistance mechanism. These results demonstrate the varied challenges presented by a natural virus population, and support the notion that the CRISPR/viritope system must be able to adapt quickly to provide host immunity. The ability of metagenomics to track population-level variation in viritope sequences allows for a culture independent method for evaluating the fast co-evolution of host and viral genomes and its consequence on the structuring of complex microbial communities. PMID- 19132094 TI - High-intensity focused ultrasound in small renal masses. AB - High-intensity focused ultrasound (HIFU) competes with radiofrequency and cryotherapy for the treatment of small renal masses as a third option among ablative approaches. As an emerging technique, its possible percutaneous or laparoscopic application, low discomfort to the patient and the absence of complications make this technology attractive for the management of small renal masses. This manuscript will focus on the principles, basic research and clinical applications of HIFU in small renal masses, reviewing the present literature. Therapeutic results are controversial and from an clinical view, HIFU must be considered a technique under investigation at present time. Further research is needed to settle its real indications in the management of small renal masses; maybe technical improvements will certainly facilitate its use in the management of small renal masses in the near future. PMID- 19132095 TI - Security Framework for Pervasive Healthcare Architectures Utilizing MPEG-21 IPMP Components. AB - Nowadays in modern and ubiquitous computing environments, it is imperative more than ever the necessity for deployment of pervasive healthcare architectures into which the patient is the central point surrounded by different types of embedded and small computing devices, which measure sensitive physical indications, interacting with hospitals databases, allowing thus urgent medical response in occurrences of critical situations. Such environments must be developed satisfying the basic security requirements for real-time secure data communication, and protection of sensitive medical data and measurements, data integrity and confidentiality, and protection of the monitored patient's privacy. In this work, we argue that the MPEG-21 Intellectual Property Management and Protection (IPMP) components can be used in order to achieve protection of transmitted medical information and enhance patient's privacy, since there is selective and controlled access to medical data that sent toward the hospital's servers. PMID- 19132093 TI - Identification of pax6-dependent gene regulatory networks in the mouse lens. AB - Lineage-specific DNA-binding transcription factors regulate development by activating and repressing particular set of genes required for the acquisition of a specific cell type. Pax6 is a paired domain and homeodomain-containing transcription factor essential for development of central nervous, olfactory and visual systems, as well as endocrine pancreas. Haploinsufficiency of Pax6 results in perturbed lens development and homeostasis. Loss-of-function of Pax6 is incompatible with lens lineage formation and results in abnormal telencephalic development. Using DNA microarrays, we have identified 559 genes expressed differentially between 1-day old mouse Pax6 heterozygous and wild type lenses. Of these, 178 (31.8%) were similarly increased and decreased in Pax6 homozygous embryonic telencephalon [Holm PC, Mader MT, Haubst N, Wizenmann A, Sigvardsson M, Gotz M (2007) Loss- and gain-of-function analyses reveals targets of Pax6 in the developing mouse telencephalon. Mol Cell Neurosci 34: 99-119]. In contrast, 381 (68.2%) genes were differently regulated between the lens and embryonic telencephalon. Differential expression of nine genes implicated in lens development and homeostasis: Cspg2, Igfbp5, Mab21l2, Nrf2f, Olfm3, Spag5, Spock1, Spon1 and Tgfb2, was confirmed by quantitative RT-PCR, with five of these genes: Cspg2, Mab21l2, Olfm3, Spag5 and Tgfb2, identified as candidate direct Pax6 target genes by quantitative chromatin immunoprecipitation (qChIP). In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins. This led to the identification of two and three sites in the respective Mab21l2 and Tgfb2 promoter regions identified by qChIPs. Collectively, the present studies represent an integrative genome-wide approach to identify downstream networks controlled by Pax6 that control mouse lens and forebrain development. PMID- 19132096 TI - Performance Evaluation of an Enhanced Uplink 3.5G System for Mobile Healthcare Applications. AB - The present paper studies the prospective and the performance of a forthcoming high-speed third generation (3.5G) networking technology, called enhanced uplink, for delivering mobile health (m-health) applications. The performance of 3.5G networks is a critical factor for successful development of m-health services perceived by end users. In this paper, we propose a methodology for performance assessment based on the joint uplink transmission of voice, real-time video, biological data (such as electrocardiogram, vital signals, and heart sounds), and healthcare records file transfer. Various scenarios were concerned in terms of real-time, nonreal-time, and emergency applications in random locations, where no other system but 3.5G is available. The accomplishment of quality of service (QoS) was explored through a step-by-step improvement of enhanced uplink system's parameters, attributing the network system for the best performance in the context of the desired m-health services. PMID- 19132097 TI - Neuronal plasticity in the entorhinal cortex. PMID- 19132098 TI - Endoscopic management of upper tract urothelial carcinoma. AB - Nephroureterectomy is currently the gold standard for management of upper urinary tract urothelial carcinoma despite it results. This review article in the loss of a renal unit. The ultimate aim of endoscopic management of this condition is cancer control whilst preserving renal function and the integrity of the urinary tract. Endoscopic treatments of upper tract TCC include the antegrade percutaneous and retrograde ureteroscopic approaches. This review article summarizes the endoscopic management of upper tract urothelial carcinoma, surveillance of the disease after endoscopic management and adjuvant therapy. The main message regarding endoscopic management of upper tract urothelial cancer is that patients must be carefully selected. Patient selection is based on tumour size, grade, and multifocality. Single low-grade tumours, less than 1.5 cm in size, generally have a good outcome with endoscopic treatment provided that they have regular ureteroscopic surveillance. Ureteroscopic treatment of high-grade tumours is essentially palliative. It is essential that patients are motivated and compliant as lifetime follow-up is necessary. However, until large randomized trials with long-term follow-up are performed, endoscopic management cannot be considered a standard treatment and should be limited to poor performance status patients. PMID- 19132099 TI - The characterization of high-fat diet and multiple low-dose streptozotocin induced type 2 diabetes rat model. AB - AIM: Based on the previously established method, we developed a better and stable animal model of type 2 diabetes mellitus by high-fat diet combined with multiple low-dose STZ injections. Meanwhile, this new model was used to evaluate the antidiabetic effect of berberine. METHOD: Wistar male rats fed with regular chow for 4 weeks received vehicle (control groups), rats fed with high-fat diet for 4 weeks received different amounts of STZ once or twice by intraperitoneal injection (diabetic model groups), and diabetic rats were treated with berberine (100 mg/kg, berberine treatment group). Intraperitoneal glucose tolerance test and insulin tolerance test were carried out. Moreover, fasting blood glucose, fasting insulin, total cholesterol, and triglyceride were measured to evaluate the dynamic blood sugar and lipid metabolism. RESULT: The highest successful rate (100%) was observed in rats treated with a single injection of 45 mg/kg STZ, but the plasma insulin level of this particular group was significantly decreased, and ISI has no difference compared to control group. The successful rate of 30 mg/kg STZ twice injection group was significantly high (85%) and the rats in this group presented a typical characteristic of T2DM as insulin resistance, hyperglycemia, and blood lipid disorder. All these symptoms observed in the 30 mg/kg STZ twice injection group were recovered by the treatment of berberine. CONCLUSION: Together, these results indicated that high-fat diet combined with multiple low doses of STZ (30 mg/kg at weekly intervals for 2 weeks) proved to be a better way for developing a stable animal model of type 2 diabetes, and this new model may be suitable for pharmaceutical screening. PMID- 19132100 TI - Role of visible light-activated photocatalyst on the reduction of anthrax spore induced mortality in mice. AB - BACKGROUND: Photocatalysis of titanium dioxide (TiO(2)) substrates is primarily induced by ultraviolet light irradiation. Anion-doped TiO(2) substrates were shown to exhibit photocatalytic activities under visible-light illumination, relative environmentally-friendly materials. Their anti-spore activity against Bacillus anthracis, however, remains to be investigated. We evaluated these visible-light activated photocatalysts on the reduction of anthrax spore-induced pathogenesis. METHODOLOGY/PRINCIPAL FINDINGS: Standard plating method was used to determine the inactivation of anthrax spore by visible light-induced photocatalysis. Mouse models were further employed to investigate the suppressive effects of the photocatalysis on anthrax toxin- and spore-mediated mortality. We found that anti-spore activities of visible light illuminated nitrogen- or carbon doped titania thin films significantly reduced viability of anthrax spores. Even though the spore-killing efficiency is only approximately 25%, our data indicate that spores from photocatalyzed groups but not untreated groups have a less survival rate after macrophage clearance. In addition, the photocatalysis could directly inactivate lethal toxin, the major virulence factor of B. anthracis. In agreement with these results, we found that the photocatalyzed spores have tenfold less potency to induce mortality in mice. These data suggest that the photocatalysis might injury the spores through inactivating spore components. CONCLUSION/SIGNIFICANCE: Photocatalysis induced injuries of the spores might be more important than direct killing of spores to reduce pathogenicity in the host. PMID- 19132101 TI - Predicting the herd immunity threshold during an outbreak: a recursive approach. AB - BACKGROUND: The objective was to develop a novel algorithm that can predict, based on field survey data, the minimum vaccination coverage required to reduce the mean number of infections per infectious individual to less than one (the Outbreak Response Immunization Threshold or ORIT) from up to six days in the advance. METHODOLOGY/PRINCIPAL FINDINGS: First, the relationship between the rate of immunization and the ORIT was analyzed to establish a link. This relationship served as the basis for the development of a recursive algorithm that predicts the ORIT using survey data from two consecutive days. The algorithm was tested using data from two actual measles outbreaks. The prediction day difference (PDD) was defined as the number of days between the second day of data input and the day of the prediction. The effects of different PDDs on the prediction error were analyzed, and it was found that a PDD of 5 minimized the error in the prediction. In addition, I developed a model demonstrating the relationship between changes in the vaccination coverage and changes in the individual reproduction number. CONCLUSIONS/SIGNIFICANCE: The predictive algorithm for the ORIT generates a viable prediction of the minimum number of vaccines required to stop an outbreak in real time. With this knowledge, the outbreak control agency may plan to expend the lowest amount of funds required stop an outbreak, allowing the diversion of the funds saved to other areas of medical need. PMID- 19132102 TI - Common and distinct genetic properties of ESCRT-II components in Drosophila. AB - BACKGROUND: Genetic studies in yeast have identified class E vps genes that form the ESCRT complexes required for protein sorting at the early endosome. In Drosophila, mutations of the ESCRT-II component vps25 cause endosomal defects leading to accumulation of Notch protein and increased Notch pathway activity. These endosomal and signaling defects are thought to account for several phenotypes. Depending on the developmental context, two different types of overgrowth can be detected. Tissue predominantly mutant for vps25 displays neoplastic tumor characteristics. In contrast, vps25 mutant clones in a wild-type background trigger hyperplastic overgrowth in a non-autonomous manner. In addition, vps25 mutant clones also promote apoptotic resistance in a non autonomous manner. PRINCIPAL FINDINGS: Here, we genetically characterize the remaining ESCRT-II components vps22 and vps36. Like vps25, mutants of vps22 and vps36 display endosomal defects, accumulate Notch protein and--when the tissue is predominantly mutant--show neoplastic tumor characteristics. However, despite these common phenotypes, they have distinct non-autonomous phenotypes. While vps22 mutations cause strong non-autonomous overgrowth, they do not affect apoptotic resistance. In contrast, vps36 mutations increase apoptotic resistance, but have little effect on non-autonomous proliferation. Further characterization reveals that although all ESCRT-II mutants accumulate Notch protein, only vps22 and vps25 mutations trigger Notch activity. CONCLUSIONS/SIGNIFICANCE: The ESCRT II components vps22, vps25 and vps36 display common and distinct genetic properties. Our data redefine the role of Notch for hyperplastic and neoplastic overgrowth in these mutants. While Notch is required for hyperplastic growth, it appears to be dispensable for neoplastic transformation. PMID- 19132103 TI - A novel form of memory for auditory fear conditioning at a low-intensity unconditioned stimulus. AB - Fear is one of the most potent emotional experiences and is an adaptive component of response to potentially threatening stimuli. On the other hand, too much or inappropriate fear accounts for many common psychiatric problems. Cumulative evidence suggests that the amygdala plays a central role in the acquisition, storage and expression of fear memory. Here, we developed an inducible striatal neuron ablation system in transgenic mice. The ablation of striatal neurons in the adult brain hardly affected the auditory fear learning under the standard condition in agreement with previous studies. When conditioned with a low intensity unconditioned stimulus, however, the formation of long-term fear memory but not short-tem memory was impaired in striatal neuron-ablated mice. Consistently, the ablation of striatal neurons 24 h after conditioning with the low-intensity unconditioned stimulus, when the long-term fear memory was formed, diminished the retention of the long-term memory. Our results reveal a novel form of the auditory fear memory depending on striatal neurons at the low-intensity unconditioned stimulus. PMID- 19132104 TI - Relationship between expression of the family of M proteins and lipoteichoic acid to hydrophobicity and biofilm formation in Streptococcus pyogenes. AB - BACKGROUND: Hydrophobicity is an important attribute of bacteria that contributes to adhesion and biofilm formation. Hydrophobicity of Streptococcus pyogenes is primarily due to lipoteichoic acid (LTA) on the streptococcal surface but the mechanism(s) whereby LTA is retained on the surface is poorly understood. In this study, we sought to determine whether members of the M protein family consisting of Emm (M protein), Mrp (M-related protein), Enn (an M-like protein), and the streptococcal protective antigen (Spa) are involved in anchoring LTA in a manner that contributes to hydrophobicity of the streptococci and its ability to form biofilms. METHODOLOGY/PRINCIPAL FINDINGS: Isogenic mutants defective in expression of emm, mrp, enn, and/or spa genes of eight different serotypes and their parental strains were tested for differences in LTA bound to surface proteins, LTA released into the culture media, and membrane-bound LTA. The effect of these mutations on the ability of streptococci to form a hydrophobic surface and to generate biofilms was also investigated. A recombinant strain overexpressing Emm1 was also engineered and similarly tested. The serotypes tested ranged from those that express only a single M protein gene to those that express two or three members of the M protein family. Overexpression of Emm1 led to enhanced hydrophobicity and biofilm formation. Inactivation of emm in those serotypes expressing only a single emm gene reduced biofilm formation, and protein-bound LTA on the surface, but did not alter the levels of membrane-bound LTA. The results were more varied in those serotypes that express two to three members of the M protein family. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that the formation of complexes with members of the M protein family is a common mechanism for anchoring LTA on the surface in a manner that contributes to hydrophobicity and to biofilm formation in S. pyogenes, but these activities in some serotypes are dependent on a trypsin-sensitive protein(s) that remains to be identified. The need for interactions between LTA and M proteins may impose functional constraints that limit variations in the sequence of the M proteins, major virulence factors of S. pyogenes. PMID- 19132105 TI - Variations in the electrostatic landscape of class II human leukocyte antigen molecule induced by modifications in the myelin basic protein peptide: a theoretical approach. AB - The receptor-ligand interactions involved in the formation of the complex between Class II Major Histocompatibility Complex molecules and antigenic peptides, which are essential for establishing an adaptive immunological response, were analyzed in the Class II Human Leukocyte Antigen (HLA)--Myelin Basic Protein (MBP) peptide complex (HLA-DRbeta1*1501-MBP) using a multipolar molecular electrostatic potential approach. The Human Leukocyte Antigen--peptide complex system was divided into four pockets together with their respective peptide fragment and the corresponding occupying amino acid was replaced by each of the remaining 19 amino acids. Partial atomic charges were calculated by a quantum chemistry approach at the Hatree Fock/3-21*G level, to study the behavior of monopole, dipole and quadrupole electrostatic multipolar moments. Two types of electrostatic behavior were distinguished in the pockets' amino acids: "anchoring" located in Pocket 1 and 4, and "recognition" located in Pocket 4 and 7. According to variations in the electrostatic landscape, pockets were ordered as: Pocket 1>Pocket 9>>Pocket 4 approximately Pocket 7 which is in agreement with the binding ability reported for Class II Major Histocompatibility Complex pockets. In the same way, amino acids occupying the polymorphic positions beta13R, beta26F, beta28D, beta9W, beta74A, beta47F and beta57D were shown to be key for this Receptor-Ligand interaction. The results show that the multipolar molecular electrostatic potential approach is appropriate for characterizing receptor-ligand interactions in the MHC-antigenic peptide complex, which could have potential implications for synthetic vaccine design. PMID- 19132106 TI - Production of nitric oxide and expression of inducible nitric oxide synthase in ovarian cystic tumors. AB - Tumor sections from nonneoplastic (n = 15), benign (n = 28), and malignant ovarian tumors (n = 20) were obtained from 63 women. Immunohistochemistry of the tumor sections demonstrated that inducible nitric oxide synthase (iNOS) expression was increased in ovarian cancer samples compared to nonneoplastic or benign tumor samples. Using the Griess method, nitric oxide (NO) metabolite levels were also found to be elevated in malignant tumor samples compared to benign tumor samples (P < .05). For stage I ovarian cancer, intracystic NO levels >80 microM were more frequent than NO levels <80 microM, and iNOS expression in well-differentiated carcinomas was greater than in moderately/poorly differentiated carcinomas (P < .05). These data suggest an important role for NO in ovarian carcinogenesis. PMID- 19132110 TI - Editors' clubs benefit researchers and trainees. PMID- 19132113 TI - Disease modeling for Ebola and Marburg viruses. AB - The filoviruses Ebola and Marburg are zoonotic agents that are classified as both biosafety level 4 and category A list pathogens. These viruses are pathogenic in humans and cause isolated infections or epidemics of viral hemorrhagic fever, mainly in Central Africa. Their natural reservoir has not been definitely identified, but certain species of African bat have been associated with Ebola and Marburg infections. Currently, there are no licensed options available for either treatment or prophylaxis. Different animal models have been developed for filoviruses including mouse, guinea pig and nonhuman primates. The 'gold standard' animal models for pathogenesis, treatment and vaccine studies are rhesus and cynomolgus macaques. This article provides a brief overview of the clinical picture and the pathology/pathogenesis of human filovirus infections. The current animal model options are discussed and compared with regard to their value in different applications. In general, the small animal models, in particular the mouse, are the most feasible for high biocontainment facilities and they offer the most options for research owing to the greater availability of immunologic and genetic tools. However, their mimicry of the human diseases as well as their predictive value for therapeutic efficacy in primates is limited, thereby making them, at best, valuable initial screening tools for pathophysiology, treatment and vaccine studies. PMID- 19132114 TI - I kid you not. PMID- 19132115 TI - The fission yeast model for the lysosomal storage disorder Batten disease predicts disease severity caused by mutations in CLN3. AB - The function of the CLN3 protein, which is mutated in patients with the neurodegenerative lysosomal storage disorder Batten disease, has remained elusive since it was identified 13 years ago. Here, we exploited the Schizosaccharomyces pombe model to gain new insights into CLN3 function. We modelled all missense mutations of CLN3 in the orthologous protein Btn1p, as well as a series of targeted mutations, and assessed trafficking and the ability of the mutant proteins to rescue four distinct phenotypes of btn1Delta cells. Mutating the C terminal cysteine residues of Btn1p caused it to be internalised into the vacuole, providing further evidence that this protein functions from pre-vacuole compartments. Mutations in the lumenal regions of the multi-spanning membrane protein, especially in the third lumenal domain which contains a predicted amphipathic helix, had the most significant impact on Btn1p function, indicating that these domains of CLN3 are functionally important. Only one mutant protein was able to rescue the cell curving phenotype (p.Glu295Lys), and since this mutation is associated with a very protracted disease progression, this phenotype could be used to predict the disease severity of novel mutations in CLN3. The ability to predict disease phenotypes in S. pombe confirms this yeast as an invaluable tool to understanding Batten disease. PMID- 19132116 TI - Fishing for the genetic basis of cardiovascular disease. AB - Cardiovascular disease (CVD) has recently overtaken infectious disease to become the biggest global killer. Genetic factors have emerged as being of major importance in the pathogenesis of CVD. Owing to disease heterogeneity, variable penetrance and high mortality, human genetic studies alone are not sufficient to elucidate the genetic basis of CVD. Animal models are needed to identify novel genes that are involved in cardiovascular pathology and to verify the effect of suspected disease genes on cardiovascular function. An intriguing model organism is the zebrafish danio rerio. Several features of the zebrafish, such as a closed cardiovascular system, transparency at embryonal stages, rapid and external development, and easily tractable genetics make it ideal for cardiovascular research. Moreover, zebrafish are suitable for forward genetics approaches, which allow the unbiased identification of novel and unanticipated cardiovascular genes. Zebrafish mutants with various cardiovascular phenotypes that closely correlate with human disease, such as congenital heart disease, cardiomyopathies and arrhythmias, have been isolated. The pool of zebrafish mutants, for which the causal gene mutation has been identified, is constantly growing. The human orthologues of several of these zebrafish genes have been shown to be involved in the pathogenesis of human CVD. Cardiovascular zebrafish models also provide the opportunity to develop and test novel therapeutic strategies, using innovative technologies such as high throughput in vivo small molecule screens. PMID- 19132117 TI - Foci of Listeria monocytogenes persist in the bone marrow. AB - Murine listeriosis is one of the most comprehensive and well-studied models of infection, and Listeria monocytogenes has provided seminal information regarding bacterial pathogenesis. However, many aspects of the mouse model remain poorly understood, including carrier states and chronic colonization which represent important features of the spectrum of host-pathogen interaction. Bone marrow has recently been shown to harbor L. monocytogenes, which spreads from this location to the central nervous system. Bone could, therefore, be an important chronic reservoir, but this infection is difficult to study because it involves only a few bacteria and the extent of infection cannot be assessed until after the animal is sacrificed. We employed in vivo bioluminescence imaging to localize L. monocytogenes bone infections over time in live mice, revealing that the bacteria grow in discrete foci. These lesions can persist in many locations in the legs of mice and are not accompanied by a histological indication such as granuloma or a neutrophil infiltratate. We demonstrate that highly attenuated hly mutants, which have defective intracellular replication, are capable of prolonged focal infection of the bone marrow for periods of up to several weeks. These results support the recently proposed hypothesis that the bone marrow is a unique niche for L. monocytogenes. PMID- 19132118 TI - Expression of H-RASV12 in a zebrafish model of Costello syndrome causes cellular senescence in adult proliferating cells. AB - Constitutively active, 'oncogenic' H-RAS can drive proliferation and transformation in human cancer, or be a potent inducer of cellular senescence. Moreover, aberrant activation of the Ras pathway owing to germline mutations can cause severe developmental disorders. In this study we have generated transgenic zebrafish that constitutively express low levels, or can be induced to express high levels, of oncogenic H-RAS. We observed that fish carrying the integrated transgene in their germline display several hallmarks of Costello syndrome, a rare genetic disease caused by activating mutations in the gene H-RAS, and can be used as a model for the disease. In Costello-like fish, low levels of oncogenic H RAS expression are associated with both reduced proliferation and an increase in senescence markers in adult progenitor cell compartments in the brain and heart, together with activated DNA damage responses. Overexpression of H-RAS through a heat-shock-inducible promoter in larvae led to hyperproliferation, activation of the DNA damage response and tp53-dependent cell cycle arrest. Thus, oncogene induced senescence of adult proliferating cells contributes to the development of Costello syndrome and provides an alternative pathway to transformation in the presence of widespread constitutively active H-RAS expression. PMID- 19132120 TI - MDM2-A, a common Mdm2 splice variant, causes perinatal lethality, reduced longevity and enhanced senescence. AB - MDM2 is the predominant negative regulator of p53 that functions to maintain the appropriate level of expression and activity of this central tumor suppressor. Mdm2-a is a commonly identified splice variant of Mdm2; however, its physiological function is unclear. To gain insight into the activity of MDM2-A and its potential impact on p53, an Mdm2-a transgenic mouse model was generated. Mdm2-a transgenic mice displayed a homozygous-lethal phenotype that could be rescued by a reduction in p53 expression, demonstrating a dependence upon p53. Mdm2-a hemizygous mice exhibited reduced longevity, and enhanced senescence was observed in their salivary glands. In addition, the transgenic mice lacked typical, accelerated aging phenotypes. Growth of transgenic mouse embryonic fibroblasts (MEFs) was inhibited relative to wild-type MEFs, and MDM2-A was shown to bind to full-length MDM2 in an interaction that could increase p53 activity via reduced MDM2 inhibition. Evidence of p53 activation was shown in the Mdm2-a transgenic MEFs, including p53-dependent growth inhibition and elevated expression of the p53 target protein p21. In addition, MDM2-A increased senescence in a p21-independent manner. In conclusion, unexpected roles for MDM2 A in longevity and senescence were identified in a transgenic mouse model, suggesting that Mdm2 splice variants might be determinants of these phenotypes in vivo. PMID- 19132124 TI - The birth of cloning: an interview with John Gurdon. [Interview by Kristin Kain]. PMID- 19132128 TI - DMM awards travel funds to early-career researchers. PMID- 19132123 TI - Progenitor cell therapies for traumatic brain injury: barriers and opportunities in translation. AB - Traumatic brain injury (TBI) directly affects nearly 1.5 million new patients per year in the USA, adding to the almost 6 million cases in patients who are permanently affected by the irreversible physical, cognitive and psychosocial deficits from a prior injury. Adult stem cell therapy has shown preliminary promise as an option for treatment, much of which is limited currently to supportive care. Preclinical research focused on cell therapy has grown significantly over the last decade. One of the challenges in the translation of this burgeoning field is interpretation of the promising experimental results obtained from a variety of cell types, injury models and techniques. Although these variables can become barriers to a collective understanding and to evidence based translation, they provide crucial information that, when correctly placed, offers the opportunity for discovery. Here, we review the preclinical evidence that is currently guiding the translation of adult stem cell therapy for TBI. PMID- 19132125 TI - A zebrafish model for Waardenburg syndrome type IV reveals diverse roles for Sox10 in the otic vesicle. AB - In humans, mutations in the SOX10 gene are a cause of the auditory-pigmentary disorder Waardenburg syndrome type IV (WS4) and related variants. SOX10 encodes an Sry-related HMG box protein essential for the development of the neural crest; deafness in WS4 and other Waardenburg syndromes is usually attributed to loss of neural-crest-derived melanocytes in the stria vascularis of the cochlea. However, SOX10 is strongly expressed in the developing otic vesicle and so direct roles for SOX10 in the otic epithelium might also be important. Here, we examine the otic phenotype of zebrafish sox10 mutants, a model for WS4. As a cochlea is not present in the fish ear, the severe otic phenotype in these mutants cannot be attributed to effects on this tissue. In zebrafish sox10 mutants, we see abnormalities in all otic placodal derivatives. Gene expression studies indicate deregulated expression of several otic genes, including fgf8, in sox10 mutants. Using a combination of mutant and morphant data, we show that the three sox genes belonging to group E (sox9a, sox9b and sox10) provide a link between otic induction pathways and subsequent otic patterning: they act redundantly to maintain sox10 expression throughout otic tissue and to restrict fgf8 expression to anterior macula regions. Single-cell labelling experiments indicate a small and transient neural crest contribution to the zebrafish ear during normal development, but this is unlikely to account for the strong defects seen in the sox10 mutant. We discuss the implication that the deafness in WS4 patients with SOX10 mutations might reflect a haploinsufficiency for SOX10 in the otic epithelium, resulting in patterning and functional abnormalities in the inner ear. PMID- 19132131 TI - Peroxisome proliferator-activated receptors and hepatitis C virus-induced insulin resistance. AB - Insulin resistance and type 2 diabetes are associated with hepatitis C virus infection. A wealth of clinical and experimental data suggests that the virus is directly interfering with the insulin signalling in hepatocytes. In the case of at least one viral genotype (the type 3a), insulin resistance seems to be directly mediated by the downregulation of the peroxisome proliferator-activated receptor gamma. Whether and how this interaction may be manipulated pharmacologically, in order to improve the responsiveness to antivirals of insulin resistant chronic hepatitis C, patients remain to be fully explored. PMID- 19132132 TI - Mouse genome-wide association mapping needs linkage analysis to avoid false positive Loci. AB - We carried out genome-wide association (GWA) studies in inbred mouse strains characterized for their lung tumor susceptibility phenotypes (spontaneous or urethane-induced) with panels of 12,959 (13K) or 138,793 (140K) single-nucleotide polymorphisms (SNPs). Above the statistical thresholds, we detected only SNP rs3681853 on Chromosome 5, two SNPs in the pulmonary adenoma susceptibility 1 (Pas1) locus, and SNP rs4174648 on Chromosome 16 for spontaneous tumor incidence, urethane-induced tumor incidence, and urethane-induced tumor multiplicity, respectively, with the 13K SNP panel, but only the Pas1 locus with the 140K SNP panel. Haplotype analysis carried out in the latter panel detected four additional loci. Loci reported in previous GWA studies failed to replicate. Genome-wide genetic linkage analysis in urethane-treated (BALB/cxC3H/He)F2, (BALB/cxSWR/J)F2, and (A/JxC3H/He)F2 mice showed that Pas1, but none of the other loci detected previously or herein by GWA, had a significant effect. The Lasc1 gene, identified by GWA as a functional element (Nat. Genet., 38:888-95, 2006), showed no genetic effects in the two independent intercross mouse populations containing both alleles, nor was it expressed in mouse normal lung or lung tumors. Our results indicate that GWA studies in mouse inbred strains can suffer a high rate of false-positive results and that such an approach should be used in conjunction with classical linkage mapping in genetic crosses. PMID- 19132133 TI - PPAR-delta in Vascular Pathophysiology. AB - Peroxisome proliferator-activated receptors belong to the superfamily of ligand dependent nuclear receptor transcription factors, which include three subtypes: PPAR-alpha, beta/delta, and gamma. PPAR-delta, play important roles in the regulation of cell growth and differentiation as well as tissue wound and repair. Emerging evidence has also demonstrated that PPAR-delta is implicated in lipids and glucose metabolism. Most recently, the direct effects of PPAR-delta on cardiovascular processes such as endothelial function and angiogenesis have also been investigated. Therefore, it is suggested that PPAR-delta may have critical roles in cardiovascular pathophysiology and is a potential target for therapeutic intervention of cardiovascular disorders such as atherosclerosis. PMID- 19132134 TI - Alternate Pacing of Border-Collision Period-Doubling Bifurcations. AB - Unlike classical bifurcations, border-collision bifurcations occur when, for example, a fixed point of a continuous, piecewise C1 map crosses a boundary in state space. Although classical bifurcations have been much studied, border collision bifurcations are not well understood. This paper considers a particular class of border-collision bifurcations, i.e., border-collision period-doubling bifurcations. We apply a subharmonic perturbation to the bifurcation parameter, which is also known as alternate pacing, and we investigate the response under such pacing near the original bifurcation point. The resulting behavior is characterized quantitatively by a gain, which is the ratio of the response amplitude to the applied perturbation amplitude. The gain in a border-collision period-doubling bifurcation has a qualitatively different dependence on parameters from that of a classical period-doubling bifurcation. Perhaps surprisingly, the differences are more readily apparent if the gain is plotted vs. the perturbation amplitude (with the bifurcation parameter fixed) than if plotted vs. the bifurcation parameter (with the perturbation amplitude fixed). When this observation is exploited, the gain under alternate pacing provides a useful experimental tool to identify a border-collision period-doubling bifurcation. PMID- 19132135 TI - Refined Synthesis of 2,3,4,5-Tetrahydro-1,3,3-trimethyldipyrrin, a Deceptively Simple Precursor to Hydroporphyrins. AB - 2,3,4,5-Tetrahydro-1,3,3-trimethyldipyrrin (1) is a crucial building block in the rational synthesis of chlorins and oxochlorins. The prior 5-step synthesis of 1 from pyrrole-2-carboxaldehyde (2) employed relatively simple and well-known reactions yet suffered from several drawbacks, including limited scale (>/= 0.5 g of 1 per run). A streamlined preparation of 1 has been developed that entails four steps: (i) nitro-aldol condensation of 2 and nitromethane under neat conditions to give 2-(2-nitrovinyl)pyrrole (3), (ii) reduction of 3 with NaBH(4) to give 2-(2-nitroethyl)pyrrole (4), (iii) Michael addition of 4 with mesityl oxide under neat conditions or at high concentration to give gamma nitrohexanonepyrrole 5, and (iv) reductive cyclization of 5 with zinc/ammonium formate to give 1. Several multistep transformations have been established, including the direct conversion of 2 --> 1. The advantages of the new procedures include (1) fewer steps, (2) avoidance of several problematic reagents, (3) diminished consumption of solvents and reagents, (4) lessened reliance on chromatography, and (5) scalability. The new procedures facilitate the preparation of 1 at the multigram scale. PMID- 19132136 TI - MODELING THE INTONATION OF DISCOURSE SEGMENTS FOR IMPROVED ONLINE DIALOG ACT TAGGING. AB - Prosody is an important cue for identifying dialog acts. In this paper, we show that modeling the sequence of acoustic-prosodic values as n-gram features with a maximum entropy model for dialog act (DA) tagging can perform better than conventional approaches that use coarse representation of the prosodic contour through acoustic correlates of prosody. We also propose a discriminative framework that exploits preceding context in the form of lexical and prosodic cues from previous discourse segments. Such a scheme facilitates online DA tagging and offers robustness in the decoding process, unlike greedy decoding schemes that can potentially propagate errors. Using only lexical and prosodic cues from 3 previous utterances, we achieve a DA tagging accuracy of 72% compared to the best case scenario with accurate knowledge of previous DA tag, which results in 74% accuracy. PMID- 19132137 TI - Macrocyclic Binucleating beta-Diketiminate Ligands and their Lithium, Aluminum, and Zinc Complexes. AB - The incorporation of rigid aromatic linkers into beta-diketiminate ligands creates a binucleating scaffold that holds two metals near each other. This paper discloses the synthesis, characterization, and reactivity of mBin(2-), which has a meta-substituted xylylene spacer, and pBin(2-), which has a para-substituted xylylene spacer. Lithium, aluminum, and zinc complexes of each ligand are isolated, and in some cases are characterized by X-ray crystallography. The lithium complexes are coordinated to solvent-derived THF ligands, while the zinc and aluminum complexes have alkyl ligands. Complexes of the mBin(2-) ligand have an anti conformation in which the metals are on opposite sides of the macrocycle, while pBin(2-) complexes prefer a syn conformation. The (1)H NMR spectra of the complexes demonstrate that the conformations rapidly interconvert in the lithium complexes, and less rapidly in the zinc and aluminum complexes. PMID- 19132138 TI - Moral Enhancement. AB - Opponents of biomedical enhancement often claim that, even if such enhancement would benefit the enhanced, it would harm others. But this objection looks unpersuasive when the enhancement in question is a moral enhancement - an enhancement that will expectably leave the enhanced person with morally better motives than she had previously. In this article I (1) describe one type of psychological alteration that would plausibly qualify as a moral enhancement, (2) argue that we will, in the medium-term future, probably be able to induce such alterations via biomedical intervention, and (3) defend future engagement in such moral enhancements against possible objections. My aim is to present this kind of moral enhancement as a counter-example to the view that biomedical enhancement is always morally impermissible. PMID- 19132139 TI - Toward the Synthesis of Antascomicin B. Synthesis of a Model of the C22-C34 Fragment via Ireland-Claisen and Allylic Diazene Rearrangements. AB - The C22-C34 fragment of antascomicin B lacking the C31 and C32 hydroxyl groups has been prepared in 11 steps from commercially available 2-OH-cyclohexanone. An Ireland-Claisen rearrangement was employed to install the C26 and C27 stereocenters. Our recently reported diastereoselective acyclic 1,3-reductive transposition was used to establish the remote C23 stereocenter. Directed hydrogenation was employed to set the C29 stereocenter. The model compound contains 5 of the stereocenters and all of the carbons of the corresponding fragment of antascomicin B. PMID- 19132141 TI - Decision analysis for the evaluation of diagnostic tests, prediction models and molecular markers. AB - The traditional statistical approach to the evaluation of diagnostic tests, prediction models and molecular markers is to assess their accuracy, using metrics such as sensitivity, specificity and the receiver-operating characteristic curve. However, there is no obvious association between accuracy and clinical value: it is unclear, for example, just how accurate a test needs to be in order for it to be considered "accurate enough" to warrant its use in patient care. Decision analysis aims to assess the clinical value of a test by assigning weights to each possible consequence. These methods have been historically considered unattractive to the practicing biostatistician because additional data from the literature, or subjective assessments from individual patients or clinicians, are needed in order to assign weights appropriately. Decision analytic methods are available that can reduce these additional requirements. These methods can provide insight into the consequences of using a test, model or marker in clinical practice. PMID- 19132142 TI - A NOVEL ALGORITHM FOR UNSUPERVISED PROSODIC LANGUAGE MODEL ADAPTATION. AB - Symbolic representations of prosodic events have been shown to be useful for spoken language applications such as speech recognition. However, a major drawback with categorical prosody models is their lack of scalability due to the difficulty in annotating large corpora with prosodic tags for training. In this paper, we present a novel, unsupervised adaptation technique for bootstrapping categorical prosodic language models (PLMs) from a small, annotated training set. Our experiments indicate that the adaptation algorithm significantly improves the quality and coverage of the PLM. On a test set derived from the Boston University Radio News corpus, the adapted PLM gave a relative improvement of 13.8% over the seed PLM on the binary pitch accent detection task, while reducing the OOV rate by 16.5% absolute. PMID- 19132140 TI - New sources and instrumentation for neutrons in biology. AB - Neutron radiation offers significant advantages for the study of biological molecular structure and dynamics. A broad and significant effort towards instrumental and methodological development to facilitate biology experiments at neutron sources worldwide is reviewed. PMID- 19132143 TI - Occasional Reinforced Responses During Extinction Can Slow the Rate of Reacquisition of an Operant Response. AB - Three experiments with rats examined reacquisition of an operant response after either extinction or a response-elimination procedure that included occasional reinforced responses during extinction. In each experiment, reacquisition was slower when response elimination had included occasional reinforced responses, although the effect was especially evident when responding was examined immediately following each response-reinforcer pairing during reacquisition (Experiments 2 and 3). An extinction procedure with added noncontingent reinforcers also slowed reacquisition (Experiment 3). The results are consistent with research in classical conditioning (Bouton, Woods, & Pineno, 2004) and suggest that rapid reacquisition after extinction is analogous to a renewal effect that occurs when reinforced responses signal a return to the conditioning context. Clinical implications are also discussed. PMID- 19132144 TI - Off-Pump Coronary Artery Bypass Leads to a Regional Hypercoagulable State Not Detectable Using Systemic Markers. AB - OBJECTIVE: It is believed that off-pump coronary artery bypass grafting (OPCAB) leads to hypercoagulability, but efforts to document such a state have been unrevealing. We hypothesized that OPCAB increases the risk of developing a regional hypercoagulable state. METHODS: Blood was obtained from the aorta and coronary sinus (CS) after CABG performed off- (N=69) or on-pump (N=35) to determine the transcardiac gradients of F1.2 (thrombin production), XIIa (coagulation activation), myoglobin (ischemia) and IL-6, IL-8 using ELISA and platelet-derived microparticles using FACS. Platelet function was measured using aggregometry. Regional myocardial pH and SVG flow were recorded intraoperatively. SVG biopsies were analyzed for endothelial integrity (EI) using immunohistochemistry and graft patency was determined by predischarge CT angiography. RESULTS: Compared with on-pump, OPCAB provoked significantly higher transcardiac F1.2 (117+/-200 v. 31+/-38%), FXII-a (14+/-29 v. 2+/-4%), microparticles (14+/--9.5% v. 6.4+/--4.1%), IL-6 (119+/-183 v. 28+/-39%), and a trend toward increased IL-8 (67+/-94 v. 24+/-46%, P = 0.077). Myoglobin release after OPCAB, also greater than on-pump CABG (54+/-89 v. 8+/-14%, P < 0.01), correlated with regional pH change (R=-0.96, P < 0.0001), and F1.2 release (R=0.55, P = 0.0002). In contrast, systemic changes in these markers were all less after OPCAB. SVG flow was significantly reduced in OPCAB (39.4 versus 66.5 mL/min, P = 0.0002), but EI and graft patency rates were the same. CONCLUSIONS: Through the use of transcardiac assays, we illustrated that regional coagulation was enhanced after off- compared with on-pump CABG. If the findings of this pilot study are confirmed, OPCAB may require additional antithrombotic therapies to respond to this local hypercoagulable state. PMID- 19132146 TI - [Magnetic resonance imaging of the joints: a revolution for the practicing rheumatologist]. AB - In the last 15 years, new imaging techniques have changed the life of practicing rheumatologists in terms of both diagnostic approach and knowledge of disease mechanisms. Clinical symptoms, disease signs and the results of physical examination have been more closely related to their anatomical basis. In particular, magnetic resonance imaging allow diagnosis of disease in its early phase and its follow-up with a previously unknown sensitivity. Novel imaging studies have contributed to elucidate several pathogenetic mechanisms in musculoskeletal diseases, such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, polymyalgia rheumatica and osteoarthritis; allow evaluation of the real degree of joint inflammation, which is often uncoupled from clinical signs; and possibly reduce the need for large clinical trials. In conclusion, new imaging techniques and refinements of the established techniques have opened exciting perspectives in our understanding and treatment of many rheumatic diseases. Much attention should be paid to the training of new generations of rheumatologists in this field. PMID- 19132145 TI - MECP2 promoter methylation and X chromosome inactivation in autism. AB - Epigenetic mechanisms have been proposed to play a role in the etiology of autism. This hypothesis is supported by the discovery of increased MECP2 promoter methylation associated with decreased MeCP2 protein expression in autism male brain. To further understand the influence of female X chromosome inactivation (XCI) and neighboring methylation patterns on aberrant MECP2 promoter methylation in autism, multiple methylation analyses were peformed on brain and blood samples from individuals with autism. Bisulfite sequencing analyses of a region 0.6 kb upstream of MECP2 in brain DNA samples revealed an abrupt transition from a highly methylated region in both sexes to a region unmethylated in males and subject to XCI in females. Chromatin immunoprecipitation analysis demonstrated that the CCTC-binding factor (CTCF) bound to this transition region in neuronal cells, consistent with a chromatin boundary at the methylation transition. Male autism brain DNA samples displayed a slight increase in methylation in this transition region, suggesting a possible aberrant spreading of methylation into the MECP2 promoter in autism males across this boundary element. In addition, autistic female brain DNA samples showed evidence for aberrant MECP2 promoter methylation as an increase in the number of bisulfite sequenced clones with undefined XCI status for MECP2 but not androgen receptor (AR). To further investigate the specificity of MECP2 methylation alterations in autism, blood DNA samples from females and mothers of males with autism were also examined for XCI skewing at AR, but no significant increase in XCI skewing was observed compared to controls. These results suggest that the aberrant MECP2 methylation in autism brain DNA samples is due to locus-specific rather than global X chromosome methylation changes. PMID- 19132147 TI - [The rehabilitative approach in rheumatoid arthritis]. AB - The rehabilitative approach for the patient with rheumatoid arthritis should be early, global and complementary to an early pharmacological therapy, in the context of a multidisciplinary approach, that should include physicians with different specialties and other health professionals. Evaluation scales assessing disability and quality of life are necessary for the rehabilitative approach. These can be classified in 2 groups: specific tools and generic tools, each evaluating different components of the health status. After the evaluation and the definition of the aims of the rehabilitation, a rehabilitative project, potentially including physical therapies, therapeutic exercises, occupational therapy and orthosis should be defined. PMID- 19132148 TI - [Nailfold capillaroscopic report: qualitative and quantitative methods]. AB - Nailfold capillaroscopy (NVC) is a simple and non-invasive method used for the assessment of patients with Raynaud's phenomenon (RP) and in the differential diagnosis of various connective tissue diseases. The scleroderma pattern abnormalities (giant capillaries, haemorrages and/or avascular areas) have a positive predictive value for the development of scleroderma spectrum disorders. Thus, an analytical approach to nailfold capillaroscopy can be useful in quantitatively and reproducibly recording various parameters. We developed a new method to assess patients with RP that is capable of predicting the 5-year transition from isolated RP to RP secondary to scleroderma spectrum disorders. This model is a weighted combination of different capillaroscopic parameters (giant capillaries, microhaemorrages, number of capillaries) that allows physicians to stratify RP patients easily using a relatively simple diagram to deduce prognosis. PMID- 19132149 TI - [Dynamic contrast-enhanced magnetic resonance imaging of the wrist in early arthritis]. AB - OBJECTIVES: MRI has been proposed as the imaging method of choice to evaluate the long-term outcome in patients with early arthritis. The role of dynamic MRI, performed at presentation, in predicting the outcome of patients with early arthritis has been addressed in the present study. METHODS: 39 patients with early arthritis, involving at least one wrist, were studied with clinical visits and laboratory investigations, every 3 months. Dynamic MRI was performed with a low-field (0.2T), extremity-dedicated machine (Artoscan, Esaote, Genova, Italy) equipped with a permanent magnet and with a dedicated hand and wrist coil. During the intravenous injection of Gd-DTPA, twenty consecutive fast images of 3 slices of the wrist were acquired. The synovial contrast enhancement ratio was calculated both as rate of early enhancement (REE) per second during the first 55" and as relative enhancement (RE) at t seconds. RESULTS: In our cohort of patients, REE and RE were significantly lower than those observed in a historical cohort of 36 patients with active rheumatoid arthritis. In univariate analysis, low RE predicted complete remission of arthritis. In multivariate analysis, fulfillment of RA criteria during follow-up was predicted by high RE. The need for immunosoppressive treatment at the end of follow-up was predicted by both low RE and high REE. CONCLUSIONS: Dynamic MRI may be used to predict several outcomes of early arthritis involving the wrist. PMID- 19132150 TI - [Septic arthritis: a 12 years retrospective study in a rheumatological university clinic]. AB - BACKGROUND: Septic arthritis is a disabling and potentially life-threatening condition that requires prompt diagnosis and treatment. The most important risk factors are joint prosthesis, pre-existing joint disease and immunosuppressive drugs. The aim of our study therefore was to revaluate all septic arthritis cases discharged from our Rheumatologic Unit in the last 12 years, to assess the risk factors, the clinical and laboratory characteristics, the causative microorganisms and its possible increase in frequency. METHODS: The medical records of 42 consecutive patients with septic arthritis discharged from our Rheumatology Unit between January 1995 and December 2006 were reviewed. The patients ranged in age from 23 to 90 and there isn't gender predominance. Septic arthritis was diagnosed based on the finding of purulent material in the joint space and/or the isolation of a bacterial pathogen from joint fluid. Demographic data, risk factors, co-morbidity, clinical manifestations, time interval between symptoms onset and diagnosis, treatment and laboratory data including serum white blood cell count, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), synovial white blood cells and culture results were analysed. We considered these parameters in the whole population and in two different age groups (< or =60, >60) and tried to determine if there was a change of microorganisms involved in septic arthritis during the years. RESULTS: Of 42 patients, 47% were aged 60 and younger. Only 10 patients were admitted to our unit before 2001. A predisposing factor was recorded in 90,5% of cases: 15 patients had rheumatoid arthritis, 8 were diabetic, 6 had seronegative arthritis, 4 had a connective tissue disease, 8 patients had a prosthetic infection and 3 were subjected recently to arthrocentesis. We found that patients aged 60 and younger were more frequently affected by joint disease and had a synovial white blood cell count lower than patients older than 60. Staphylococcus aureus caused septic arthritis in 70% of cases before 2001, and only in 35,8 % after 2001. Also, after 2001, some infections were caused by more unusual pathogens, prevalently in patients treated with TNFa inhibitors. Instead Streptococcus infections were found only in patients aged 70 and older. CONCLUSION: The incidence of bacterial arthritis has increased in the last six years and there was a modification of microorganisms involved, possibly related to a greater therapeutic aggressiveness. The increased frequency of joint disease and the use of immunosuppressive drugs in patients under the age of 60 could be responsible for a lower synovial white blood cell count in these patients. PMID- 19132151 TI - [Hypocomplementemia in systemic sclerosis]. AB - BACKGROUND: Hypocomplementemia has been detected in about 15% of unselected series of SSc patients. It constitutes one of the 10 parameters needed to evaluate the European Scleroderma Study Group (EScSG) activity index. A few studies have been so far devoted to investigate the clinical manifestations correlated with this finding. OBJECTIVE: To investigate SSc patients for hypocomplementemia and point out clinical manifestations associated with it. METHODS: 302 patients with SSc consecutively admitted to the Rheumatology Unit of the Second University of Naples were enrolled in the study. SSc patients were all investigated for sex, age, disease duration, clinical and serological subset, disease activity, organ/system severity and functional status. Patients were divided into 2 groups: normo-complementemic and hypocomplementemic (low C3 and/or C4) as measured by nephelometry. RESULTS: 252 of the 302 patients had normal complementemia; 50 (16,5%) had hypocomplementemia. Significant associations were found between hypocomplementemia and EScSG activity index (p<0.0003); functional disability (i.e. HAQ-DI >0.5)( p=0.04); and the severity of general manifestations (p<0.006); skin (p<0.0001); vascular (p<0.0001); heart (p<0.0001) and lung (p<0.0001) involvement. CONCLUSIONS: Our study confirms that hypocomplementemia occurs in patients with SSc. It resulted to be associated with disease activity, functional status and the severity of distinct disease manifestations. PMID- 19132152 TI - [Distress influence in fibromyalgia]. AB - OBJECTIVE: Fibromyalgia syndrome (FM) is a controversial chronic painful syndrome. Although the aetiology is unknown, FM is frequently correlated with stressors events. Recent studies highlighted the frequent comorbidity with anxiety and depression and a close relationship between stress and pain. METHODS: We evaluated the relevance of stressors events in 23 patients with FM (mean age: 45.7+/-7.4 SD), compared with 18 healthy controls (mean age: 41.7+/-6.4 SD) and 17 patients with dysfunctional syndrome (mean age 40.8+/-6.1). We performed the italian validated rapid assessment of the stress test (VRS) for the assessment of stress. Furthermore, we evaluated the psychological history for a semi quantitative assessment (IVAS) of the 49 stress-generating events listed in 1994's DSM-IV. RESULTS: The group of "healthy" subjects showed stress values at VRS test (mean: 7.00+/-4.65 SD) significantly lower (p=0.0001) than the patients with "dysfunctional syndrome" (mean 14.82+/-7.69 SD) and those with FM (mean 20.04+/-9.90 SD). The IVAS test, showed higher values in FM subgroup than healthy (p=0.0001) and dysfunctional syndrome (p=0,007). Also, the patients with FM showed a greater gravity to attribute to single stressors events (p=0.02). CONCLUSIONS: Our results emphasize the importance of the perceived stress among the patients with FM, and support the hypothesis that FM could be due to a psycho neuro-endocrinal response to several stressors events in patients with genetical hyperresponsiveness to stress. PMID- 19132153 TI - [Phytothermotherapy with fermenting alpine grass in knee osteoarthritis: mid-long term results]. AB - This is an observational study of the mid-long-term results of a single course of phytothermotherapy with grass baths (group A, 54 patients), of a course of usual medical care (group B, 58 patients) and of a course of physiokinesistherapy (FKT, group C, 30 patients) in knee osteoarthritis. For each group of consecutively treated patients we evaluated the Lequesne algo-functional Index, the drug consumption, the frequency of the patient-physician contacts and laboratory or radiological examinations after 10-15 days of treatment and at 3, 6, 9 and 12 months with blind telephonic follow-up. The mean Lequesne-score at basal time was 7.5+/-3.3, 11.9+/-5.3 and 11.0+/-2.7 in group A, B and C respectively. In each group this score diminished at the end of the treatment (p<0.001). At 3, 6, 9 and 12 months the score remained lower than at basal time in group A (p<0.001) and group B (p<0.01), but not in group C. Drug consumption, patient-physician contacts and lab examinations were 5 times lower in group A than in group B and group C at basal time and throughout the follow-up. The study underlines the mid long term efficacy of grass baths on both pain and functionality in knee osteoarthritis; this effect, compared to basal values, was even more evident at 3 and 6 months than that of usual medical care. FKT shows improvement only at the end of the treatment, but not long-lastingly. PMID- 19132154 TI - Efficacy and safety of anti-TNF agents in the Lombardy rheumatoid arthritis network (LORHEN). AB - Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic synovitis and bone damages, which consist of joint destruction. Clinical trials have shown that anti-tumour necrosis factor (TNF) drugs are effective in patients with rheumatoid arthritis (RA) refractory to disease-modifying antirheumatic drugs (DMARDs). At about the same time as the European approval of the third anti-TNF agent for treating rheumatoid arthritis (RA) patients, the Italian Society of Rheumatology (Societa Italiana di Reumatologia [SIR]) started a database for the registration and active follow-up of patients with RA treated with biological response modifiers. Since 1999, all patients with RA (ACR criteria) and treated with at least one dose of an anti-TNF agent at four Rheumatology Centres in Lombardy (northwest Italy) have been included in the Lombardy Rheumatology Network (LORHEN) registry in order to track the efficacy and safety of the three available TNF inhibitors during the first three years of treatment. PMID- 19132155 TI - A dermatomyositis and scleroderma overlap syndrome with a remarkable high titer of anti-exosome antibodies. AB - An overlap syndrome of dermatomyositis and scleroderma is reported. The case corresponded to a 27-year-old female with a clinical picture of 14 months evolution, characterized by proximal muscles weakness, erythematous rash in wrists, knees, ankles, Gottron sign, heliotrope periorbital rash and dysphagia. A muscle biopsy was compatible with dermatomyositis; meanwhile the skin biopsy was compatible with scleroderma. Muscle enzymes were increased. Interestingly, the antinuclear antibody determination in HEp-2 cells was positive with a remarkable titer of 81,920 exhibiting a nucleolar pattern. Anti-Jo1 antibody was negative, but anti-PM/Scl-100 positive. The patient received methylprednisolone and cyclophosphamide pulses, with gradual improvement. Present report constitutes a case of overlap dermatomyositis-scleroderma syndrome, with anti-PM/Scl autoantibodies (anti-exosome). The remarkable of this case was the exceptional high antinucleolar antibody titer. PMID- 19132156 TI - [Historical evolution of the concept of fibromyalgia: the main stages]. AB - The concept of fibromyalgia is fairly recent, by evolving from previous notions such as muscular rheumatism and fibrositis. In this concise report the main stages leading to the development of the concept of fibromyalgia are sketched out, beginning from the notions of fibrositis nodules, trigger points and myofascial pain, up to the most recent knowledge including this clinical condition in the cluster of central sensitivity syndromes. PMID- 19132157 TI - [Hemostasis and thrombosis research--an intensive course in clinical hemostasis]. PMID- 19132158 TI - [Physiology of blood coagulation and fibrinolysis: biochemistry]. AB - The principles of initiator and amplifications reactions of blood coagulation and fibrinolysis will be presented and discussed in relation to various regulatory pathways of haemostasis. In particular, cell surface-dependent activation and inhibition reactions are characteristics of multicomponent enzyme complexes that also allow the endogenous control of the haemostasis system. The understanding of these relationships in complications of haemostasis has lead to different strategies for the therapeutic intervention with pro- and anticoagulant substances. PMID- 19132159 TI - [Molecular biology and haemostasis]. AB - Molecular biological methods have become increasingly important not only in the diagnostics of inherited monogenetic diseases such as hemophilia A or B but also in the diagnostics of polygenetic diseases e.g. venous and arterial thrombosis. In haemophilia A, sequencing of the factor VIII gene has been established in addition to the analysis of the two most frequent genetic abnormalities, the inversions in intron 22 and intron 1, in several centers. Molecular testing has proved helpful to identify haemophilia patients at high risk to develop inhibitors as well as in carrier analysis. In patients with familial protein C or protein S deficiency mutation analysis contributes to the verification of the diagnosis. The frequently performed tests for the factor V Leiden mutation and the prothrombin 20210G>A variation can potentially support the estimation of the thrombotic risk as well as the risk of recurrence. However, any uncritical application of these genetic tests does not improve diagnostics nor does it support therapeutic decision making or counselling of the patient. Therefore, one should only do genetic tests with medical or therapeutic consequences. The applicability of mutation analysis in the daily routine is still limited in spite of the importance of molecular diagnostics in the understanding of pathomechanisms of haemostatic disorders. As has been demonstrated in large studies, the phenotypic effects of mutations can vary significantly between individuals. Endogenous and exogenous modulators that are still largely unknown, play a role. Currently, the understanding of these modulators is limited, and large multicenter studies and meta-analyses are needed for a better understanding of gene-gene and gene-environment interactions. PMID- 19132160 TI - [Platelets: biochemistry and physiology]. AB - This article reviews the roles of blood platelets in haemostasis as well as in the pathogenesis of thromboembolic diseases. Besides the basic processes in primary haemostasis, platelet adhesion, platelet secretion, platelet aggregation, clot retraction, the new model of thrombin formation on the platelet surface is presented. The different signal transduction pathways in platelets are a main focus of this review. PMID- 19132161 TI - [Acquired platelet function disorders: pathogenesis, classification, frequency, diagnosis, clinical management]. AB - Given the high consumption of pharmacological agents in western societies, it is not surprising at all that drugs represent the most common cause of acquired platelet dysfunction. While acetylsalicylic acid, clopigogrel and integrin alphaIIbbeta3 (GPIIb-IIIa) receptor antagonists are well-known as prototypes of antiplatelet drugs, other widely used agents including non-steroidal anti inflammatory drugs, antibiotics, serotonin reuptake inhibitors, and volume expanders can also impair platelet function and cause or aggravate haemorrhages. Besides pharmacological agents, certain clinical conditions are often associated with qualitative platelet disorders and bleeding diathesis. Consequently, in contrast to inherited platelet disorders, acquired platelet function defects are much more frequent in clinical practice and deserve special attention. Their pathogenesis is widespread and heterogeneous with various, sometimes overlapping abnormalities. Moreover, acquired platelet dysfunctions can occur at any age and range in severity from mild to life-threatening haemorrhages. Due to their heterogeneity, acquired platelet function disorders will be classified and discussed according to the underlying clinical setting or disease. PMID- 19132162 TI - [Inborn and acquired von Willebrand disease]. AB - Von Willebrand disease (VWD) is known for its marked heterogeneity which was already recognized by von Willebrand in 1926. The basis of phenotypic differentiation are quantitative and qualitative or functional differences between the different types and subtypes of VWD. Clinical relevant facts for the practioner on diagnosis and therapy of von Willebrand disease and von Willebrand syndrome are presented. PMID- 19132163 TI - [Diagnostic of blood coagulation]. AB - A survey is given on the peculiar characteristics of laboratory methods for analyzing the blood coagulation system with special regard to the preanalytical, analytical and postanalytical phase. Routinely used methods are described. PMID- 19132164 TI - [Congenital coagulopathies and coagulation factor inhibitors]. AB - An overview is given on the clinical picture and therapy of the haemophilias A and B as an example for congenital coagulopathies. The survey deals with the special problems of today's "treated haemophilia" and its recommendations on diagnosis and treatment regimens. A special focus is put on the pathogenesis of the haemophilic joint arthropathy, that is affecting most haemophiliacs earlier or later during life. Safe plasma derived and recombinant clotting factor concentrates allow an efficient on-demand and also a prophylactic treatment for the prevention of bleeding sequelae. As a consequence life quality and life expectancy of today born haemophiliacs have nearly normalised. The problems of haemophilia with inhibitors and future therapeutic options are discussed. The development of inhibitors at start of therapy with factor VIII concentrates represent the nowadays most serious complication of haemophilia treatment. The only way to a long lasting eradication of the inhibitor ist the induction of immune tolerance that can be achieved by various protocols. A successful immune tolerance therapy (ITT) is the precondition for a regular prophylactic treatment regime. The Guidelines of the Federal Chamber of Physicians, the German recommendations for ITT and the consensus recommendations of an international working group provide an important lead for the conduction of immune tolerance therapy. PMID- 19132165 TI - [Acquired coagulopathies]. AB - Acquired coagulopathies regularly result from underlying diseases such as liver failure or sepsis. Optimal treatment of these underlying diseases defines longterm prognosis. In a few coagulopathies, such as immunocoagulopathies, in a relevant portion of patients an underlying disease cannot be identified. Therapeutic modulation of haemostasis aiming at the prevention of fatal bleeding or thromboembolism is able to improve the prognosis of patients with acquired coagulopathies. Most often this is the result of individualized, lab-controlled treatments, which are difficult to study. Within the last years the role of therapeutic modulation of haemostasis to reduce mortality was successfully studied for the first time in the frame of randomized placebo-controlled phase III studies. PMID- 19132166 TI - [Thrombotic microangiopathies]. AB - Thrombotic microangiopathies are disorders of microvascular occlusion and are characterized by thrombocytopenia and microangiopathic haemolytic anaemia. Varying additional signs and symptoms of organ ischaemia may be present. Thrombotic microangiopathies are pathophysiologically heterogenous disorders and include primarily thrombotic thrombocytopenic purpura and haemolytic uraemic syndrome. Besides this, they may occur in association with certain diseases or drugs, or after allogeneic haematopoietic stem cell transplantation. PMID- 19132167 TI - [Update thrombophilia]. AB - A survey on definitions, epidemiology, clinical manifestations of congenital and acquired thrombophilias is given with focus on evidence-based data. Diagnostic and therapeutical strategies are presented. Selective thrombophilia screening based on previous personal and/or family history of venous thromboembolism is more cost-effective than universal screening in all patient groups. In the majority of patients with venous thrombosis the results of thrombophilia screening do not influence the duration of oral anticoagulation. The only patient population who clearly profits from thrombophilia screening in this situation are patients with a newly diagnosed antiphospholipid syndrome, because prolonged anticoagulation can avoid the high incidence of recurrence in this patient population. Because of the increased risk of venous thrombosis during pregnancy and the puerperium, thrombophilia screening is indicated in selected patients with a previous history of venous thrombosis to be able to decide whether anticoagulation is necessary when these patients become pregnant. Significant associations with early and late pregnancy loss are observed for patients with thrombophilia while it remains unknown whether these patients profit from anticoagulation with heparin or low dose aspirin during their next pregnancy since placebo-controlled multicenter trials are not yet available. PMID- 19132168 TI - [Diagnosis and treatment of venous thrombosis]. AB - In the diagnosis of deep vein thrombosis in ambulatory patients, the recommended initial steps are assessment of clinical probability (CP) and a sensitive D-dimer test. With a low CP and negative D-dimer, thrombosis can be ruled out. All other constellations require further investigation with imaging techniques. Compression ultrasonography is the first-line investigation. Low-molecular weight heparin or fondaparinux is the treatment of choice for uncomplicated venous thrombosis. Secondary prophylaxis with a vitamin K antagonist is introduced in parallel as quickly as possible. The duration of treatment depends on the exposure and predisposing factors, weighing carefully the risk of recurrence on the one hand against the risk of bleeding on the other. The danger of a post thrombotic syndrome is reduced by the immediate begin of a long lasting compression therapy. PMID- 19132169 TI - [Clotting factor concentrates]. AB - Topics of this survey are the indications and use of plasma-derived and recombinant coagulation factor concentrates. These substitution therapies need to be handled carefully by weighing up their effectiveness against the long and short term side effects in the individual patient. PMID- 19132170 TI - [Pharmacology of heparins and direct anticoagulants]. AB - For decades, the options for therapeutic anticoagulation were limited to unfractionated heparin (UFH) and vitamin K antagonists (VKA), and their well known limitations had to be accepted. With the introduction of the various LMWHs, the short- and medium-term anticoagulation could be much improved, but an alternative to VKA is still missing The heparins provided the proof of concept that FXa and thrombin represent suitable targets for therapeutic anticoagulation. Consequently, the search for new anticoagulants focuses on inhibitors of thrombin (DTI) or FXa (DXI). Apart from the VKA, the anticoagulants presently available or in an advanced stage of development can thus be divided in two classes: One are the glyco-anticoagulants with the natural sulfated glycosaminoglycans (GAGs) (UFH, LMWHs, and danaparoid) and the synthetic oligosaccharides (OS) (fondaparinux, idraparinux, and SR123781A). The other class are the xenobiotic anticoagulants, i.e. proteins and synthetic chemical entities. Die glyco anticoagulants act partially (GAGs) or exclusively (oligosaccharides) by catalysing antithrombin, whereas the xenobiotic anticoagulants are direct inhibitors of either thrombin or FXa. At present, three parenteral DTI (lepirudin, argatroban, and bivalirudin) and since March 2008 one oral DTI (dabigatran etexilate) are clinically used for limited indications. In September 2008 rivaroxaban has been approved as the first oral DXI. This review describes the development of the anticoagualants as well as the pharmacological profile of the clinically used anticoagualants. PMID- 19132171 TI - [Anticoagulation by oral treatment with vitamin K antagonists]. AB - A survey is given on pharmacology and indications for the treatment with vitamin K antagonists. The therapeutic handling and self control by the patient is described. PMID- 19132172 TI - [Fibrinolytic therapy]. AB - Fibrinolytic therapy still is a keystone in the therapeutic approach of acute occlusion in different vascular regions. However, before starting fibrinolytic therapy an evidence-based risk assessment is necessary. This article presents the most common fibrinolytic agents as well as evidences and recommendations for fibrinolytic therapy of different vascular regions. PMID- 19132173 TI - [Anticoagulant and thrombolytic agents in acute coronary syndromes]. AB - The term acute coronary syndrome (ACS) pertains to the instable and life threatening forms of a clinically manifest coronary artery disease with biochemical and/or electro-cardiographic evidence of myocyte cell death. In detail, it includes the unstable angina pectoris, the non-ST segment elevation myocardial infarction (NSTEMI) the ST segment elevation myocardial infarction (STEMI) and as well the sudden cardiac death. As early reperfusion of ischaemic myocardium is the most effective way for limiting infarct size by restoring the balance between myocardial oxygen supply and demand, it is the most important therapeutic goal to achieve early and complete antegrade flow in the occluded or restricted vessel, related with a reduction of short and longtime complications as heart failure and severe arrhythmias. It is generally accepted, that the primary percutaneous coronary intervention (PCI) is the method of choice in acute myocardial infarction (STEMI) to restore TIMI-3 blood flow in occluded coronary arteries, if this can be performed within two hours of symptom onset and by a highly specialized team. Since these requirements are only met in 20% of hospitals caring for patients with STEMI in Germany, the therapy with thrombolytic and anticoagulant agents plays still an important role. Apart from a rapid and effective prehospital primary care, it depends furthermore on a differentiated anticoagulatory and antithrombotic therapy during coronary intervention to get optimal results. PMID- 19132174 TI - [Managing perioperative haemostasis]. AB - Managing perioperative haemostasis starts with the diligently taken patient history. Unfortunately, classic global tests such as the PT and aPTT have no predictive value with regard to an acquired intra- or postoperative bleeding diathesis. However, new assays for preoperative risk stratification are in clinical development. An attribute of good perioperative haemostasis management is the early, multidisciplinary problem assessment. With a preoperatively existing anticoagulation or antiplatelet therapy, perioperative bridging therapy needs to be carefully planned as cardiovascular risk patients have an increased risk of morbidity and mortality when their current anticoagulative therapy is simply stopped. If a haemorrhagic diathesis is known, a specific therapy should be scheduled early preoperatively. When excessive intra- and postoperative bleeding occurs, point of care diagnostics can help to determine the underlying pathophysiology. A predefined validated algorithm reduces the need for blood products. To establish an evidence based approach for the use of blood components and other procoagulants in such a situation requires prospective clinical trials. The actual knowledge on the pathophysiology of such incidents (e. g. cross linking defects by use of colloids, dilutional effects of volume therapy, repeated use of FFP, of antifibrinolytics, frequency of unwanted effects) should also be considered. PMID- 19132175 TI - [Thrombophilia in pregnancy: venous thromboembolism, fetal loss, preeclampsia, intrauterine growth restriction]. AB - Women with acquired and hereditary thrombophilia are at increased risk of developing venous thromboembolism and other associated gestational vascular complications like fetal loss, preeclampsia, intrauterine growth restriction, and placental abruption during pregnancy. These complications are a major cause of maternal and fetal morbidity and mortality. In view of the data showing an association between thrombophilia and these adverse pregnancy outcomes, clinicians are increasingly using antithrombotic therapy in women at risk of these complications. Aside from recurrent pregnancy loss in antiphospholipid syndrome and prevention of venous thromboembolism, there is limited evidence on the benefit of antithrombotic interventions to guide therapy. The data in favour of antithrombotic therapy in women with hereditable thrombophilia and vascular placental complications consist predominantly of small uncontrolled trials or observational studies. Randomized, placebo-controlled trials are lacking as most patients do not accept placebo. Further randomised controlled trials are urgently required to explore this therapeutic option. PMID- 19132176 TI - [Blood coagulation disorders in children]. AB - As in adults, haemorrhagic or thrombotic events may also occur in children. The underlying reasons are inborn or acquired. Inherited disorders usually present during in early infancy. In order to interpret clinical and laboratory findings in children, it is necessary to keep in mind some specific paediatric features. This knowledge also forms the basic requirement to choose the appropriate therapy. PMID- 19132177 TI - [Haemostatic aspects in clinical oncology]. AB - The clinical link between cancer and thrombosis has been recognized by Armand Trousseau in 1865. It has now become clear that clotting activation in malignancy not only plays an important role in the evolution of venous thromboembolism (VTE) or systemic coagulation disorders such as disseminated intravascular coagulation, but that multiple components of the haemostatic and fibrinolytic systems are directly involved in tumour progression. In particular, tissue factor (TF) appears to be involved in several pathways relevant to cancer growth and metastasis. Increasing evidence emerges that haemostatic perturbances in cancer patients are, at least in part, controlled by defined genetic events in molecular tumourigenesis including activating and inactivating mutations of oncogenes and tumour suppressor genes, respectively. Long-term therapy with low-molecular weight heparin (LMWH) is considered as standard treatment for cancer-associated VTE. However, several experimental studies and clinical trials suggest that LMWH may also be beneficial as an adjunct in the treatment of patients with malignant disease. This article provides an overview on the significance, pathogenesis and treatment of cancer-related clotting disorders as well as on the cellular and molecular mechanisms, by which haemostatic components such as TF, platelets and fibrin(ogen) drive tumour progression. PMID- 19132178 TI - Face to face with basic and clinical research in Thrombosis and Haemostasis. PMID- 19132179 TI - Dabigatran etexilate for prevention of venous thromboembolism. PMID- 19132180 TI - VTE recurrence in patients with inherited deficiencies of natural anticoagulants. PMID- 19132181 TI - Pulmonary embolism diagnosis: remember the history and physical exam. PMID- 19132182 TI - Drug-eluting balloons for percutaneous coronary interventions. PMID- 19132183 TI - Another view on prasugrel. PMID- 19132184 TI - Prasugrel development - claims and achievements. AB - The priority task to develop better and safer antiplatelet agents is difficult to overestimate. In fact, oral antiplatelet agents, such as aspirin in ISIS-2 study, and clopidogrel in COMMIT mega trial in moderate doses are among the very few classes of drugs, which lead to the absolute mortality reduction benefit in patients after acute vascular thrombotic events. Prasugrel is an experimental thienopyridine, and irreversible antagonist of ADP P2Y(12) receptor leading to inhibition of platelet activation and aggregation. The recent TRITON trial, assessing head-to-head prasugrel versus standard of care clopidogrel, both on top of aspirin, reveals numerous controversies with regard to the trial design, definition of events, interpretation of the results, and suitability of the high maintenance prasugrel dose for chronic preventive human use. We critically review various aspects of prasugrel development, focusing on the discrepancies between claims and achievements. We conclude that the benefits of prasugrel are overestimated, and the risks, especially during chronic use are underestimated. Very careful maintenance-dose selection with the main focus on long-term safety profile for the new agents will become a key to success for the future oral antiplatelet drug development. PMID- 19132185 TI - Improving outcomes with antiplatelet therapies in percutaneous coronary intervention and stenting. PMID- 19132186 TI - The 8(th) American College of Chest Physicians Guidelines - a perspective on venous thromboembolism guidelines. PMID- 19132187 TI - Compensatory anti-inflammatory response syndrome. AB - The concept of 'Compensatory anti-inflammatory response syndrome' (CARS) was proposed in 1997 by Roger Bone (1941-1997) to qualify the consequences of the counter-regulatory mechanisms initiated to limit the overzealous inflammatory process in patients with infectious (sepsis) or non-infectious systemic inflammatory response syndrome (SIRS). One major consequence of CARS is the modification of the immune status that could favour the enhanced susceptibility of intensive care patients to nosocomial infections. Indeed, most animal 'two hit' models illustrate an enhanced sensitivity to infection after a first insult. However, this observation is highly dependent on the experimental procedure. Numerous functions of circulating leukocytes are altered in sepsis and SIRS patients, as well as in animal models of sepsis or SIRS. However, this is rather a reprogramming of circulating leukocytes, since there is not a global defect of the immune cells functions. Furthermore, within tissues, leukocytes are rather primed or activated than immunosuppressed. Thus, CARS may be considered as an adapted compartmentalized response with the aim to silence some acute proinflammatory genes, and to maintain the possible expression of certain genes involved in the anti-infectious process. PMID- 19132188 TI - Biobanks and the search for predictive biomarkers of local and systemic outcome in atherosclerotic disease. AB - Multiple risk factors have been associated with progression of atherosclerosis. To identify the individual patient who is at risk for disruption of a vulnerable plaque, leading to a cardiovascular event, remains a major challenge. Current screening methods, based on traditional risk factors, do not allow risk stratification on an individual level. The discovery of new biomarkers would aid in identifying specific patient groups at risk for adverse cardiovascular events due to atherosclerotic disease progression. The current definition of the vulnerable plaque, e.g. atheromatous inflammatory plaque with a thin fibrous cap, has been based on cross-sectional post-mortem studies. The predictive value of these histological characteristics of the vulnerable plaque is likely to be low, because they are also frequently observed at multiple locations in symptomatic and asymptomatic patients. The Athero-express study follows a new concept to search for the atherosclerotic patient who may suffer from adverse events. In this study, we investigate the predictive value of local plaque composition for adverse events in other vascular territories, regarding the plaque as a concentrated expression of this systemic disease. First results from this longitudinal biobank study show that the local plaque hides strong predictive value for cardiovascular events elsewhere in the vascular tree. Longitudinal biobank studies will facilitate the identification of novel local plaque markers. The search for the plaque protein signature that is predictive for adverse events might enable patient stratification that will allow individualized tailor made medicine and subsequently guide the choice for therapeutic interventions. PMID- 19132189 TI - Prothrombin amino terminal region helps protect coagulation factor Va from proteolytic inactivation by activated protein C. AB - The hypothesis that prothrombin (FII) protects coagulation factor Va (FVa) from proteolytic inactivation by activated protein C (APC) was tested using purified proteins. FII dose-dependently protected FVa from APC proteolysis under conditions where competition of proteins for binding to negatively-charged phospholipid surface was not relevant (i.e. either at high phospholipid vesicle concentrations or using soluble dicaproylphosphatidylserine at levels below its critical micellar concentration). Cleavages in FVa at both Arg(506) and Arg(306) by APC were inhibited by FII. FII did not alter the amidolytic activity of APC towards chromogenic oligopeptide substrates or inhibit FVIIIa inactivation by APC, implying that the FII-mediated protection of FVa from APC proteolysis was due to the ability of FII to inhibit protein-protein interactions between FVa and APC. FII also protected FVa from inactivation by Gla-domainless APC, ruling out a role for the APC Gla domain for these observations. To identify domains of FII responsible for the observed phenomenon, various forms or fragments of FII were employed. Biotin-Phe-ProArg-CMK-inhibited meizothrombin and fII-fragment 1*2 protected FVa from proteolysis by APC. In contrast, no significant protection of FVa from APC cleavage was observed for Gladomainless-FII, prethrombin-1, prethrombin-2, FII fragment 1 or active site inhibited-thrombin (DEGR-thrombin). Overall, these data demonstrate that the Gla domain of FII linked to kringle 1 and 2 is necessary for the ability of FII to protect FVa from APC cleavage and support the general concept that assembly of the FII activation complex (FXa*FVa*FII*lipid surface) protects FVa from APC inactivation so that the procoagulant, thrombin generating pathway can act unhindered by APC. Only following FII activation and dissociation of the FII Gla domain fragments from the FII-ase complex, can APC inactivate FVa and down-regulate thrombin generation. PMID- 19132190 TI - Polymorphisms in the protein C gene as risk factor for venous thrombosis. AB - Protein C is an important inhibitor of blood coagulation. We investigated the effect of two polymorphisms within the promoter region of the protein C gene (C/T at position 2405 and A/G at position 2418) on risk of venous thrombosis and on plasma protein C levels. In addition the combined effect of the two polymorphisms with factor V Leiden and oral contraceptive use was investigated. Previous studies on these polymorphisms were small and were not able to investigate synergistic effects. In the Multiple Environmental and Genetic Assessment of risk factors for venous thrombosis (MEGA study), protein C levels were determined in 2,043 patients with venous thrombosis and 2,857 control subjects, and the two polymorphisms in 4,285 patients and 4,863 control subjects. The CC/GG genotype was associated with the lowest protein C levels. Compared to carriers of the TT/AA genotype - a genotype associated with higher protein C levels - the risk of venous thrombosis in CC/GG carriers was 1.3-fold increased (95% confidence interval 1.09-1.48). The combination of factor V Leiden with the CC/GG genotype led to a 4.7-fold increased risk, compared to non-carriers with the TT/AA genotype. Oral contraceptive use together with the CC/GG genotype resulted in a 5.2-fold increased risk. In conclusion, the CC/GG genotype is associated with lower levels of protein C and an elevated risk of venous thrombosis compared to the TT/AA genotype. There is no clear synergistic effect of the CC/GG genotype with factor V Leiden or oral contraceptive use on thrombotic risk. PMID- 19132191 TI - A randomized evaluation of betrixaban, an oral factor Xa inhibitor, for prevention of thromboembolic events after total knee replacement (EXPERT). AB - Betrixaban is an oral direct inhibitor of factor Xa (FXa) being developed for the prevention of venous thromboembolism (VTE). Its antithrombotic effects had not been previously tested in patients. This exploratory clinical trial in the US and Canada randomized 215 patients undergoing elective total knee replacement (TKR) in a 2:2:1 ratio to receive post-operative betrixaban 15 mg or 40 mg p.o. bid or enoxaparin 30 mg s.c. q12h, respectively, for 10-14 days. The betrixaban dosage was blinded, but enoxaparin was not. Primary efficacy outcome was the incidence of VTE, consisting of deep-vein thrombosis (DVT) on mandatory unilateral (operated leg) venography, symptomatic proximal DVT, or pulmonary embolism (PE) through Day 10-14. Safety outcomes included major and clinically significant non major bleeds through 48 h after treatment. All efficacy and bleeding outcomes were adjudicated by a blinded independent central adjudication committee. Of 214 treated patients, 175 (82%) were evaluable for primary efficacy. VTE incidence was 14/70 (20%; 95% CI: 11, 31) for betrixaban 15 mg, 10/65 (15%; 95% CI: 8, 27) for betrixaban 40 mg, and 4/40 (10%; 95% CI: 3, 24) for enoxaparin. No bleeds were reported for betrixaban 15 mg, 2 (2.4%) clinically significant non-major bleeds with betrixaban 40 mg, and one (2.3%) major and two (4.6%) clinically significant non-major bleeds with enoxaparin. A dose- and concentration-dependent effect of betrixaban on inhibition of thrombin generation and anti-Xa levels was observed. Betrixaban demonstrated antithrombotic activity and appeared well tolerated in knee replacement patients at the doses studied. PMID- 19132192 TI - Efficacy and safety of dabigatran etexilate for the prevention of venous thromboembolism following total hip or knee arthroplasty. A meta-analysis. AB - Dabigatran etexilate has been investigated in three phase III trials for the prevention of venous thromboembolism (VTE). Health technology assessment agencies increasingly require meta-analyses of all relevant evidence for an intervention, if appropriate. The objective of this study was to perform a meta-analysis of efficacy and safety data for the recommended dose of dabigatran etexilate, 220 mg once daily (od), for VTE prophylaxis after total knee arthroplasty (TKA) and total hip arthroplasty (THA), and discuss the appropriateness of combining the data. Risk ratios (RR) for VTE and bleed end-points were estimated using fixed and random effects meta-analysis. Analyses were performed combining RE-MODEL and RE-NOVATE, which compared dabigatran etexilate with enoxaparin 40 mg od after TKA and THA, respectively, and also including RE-MOBILIZE, which compared dabigatran etexilate with enoxaparin 30 mg twice daily after TKA. Tests for statistical heterogeneity were performed using the Chi-squared statistic. No significant differences were detected between dabigatran etexilate and enoxaparin in any of the end-points analysed, either in the two trial analysis (all p > 0.15), or when all three trials were combined ( all p > 0.30). RRs (random effects) for the composite end-point total VTE and all-cause mortality were 0.95 [95% confidence intervals 0.82 - 1.10] and 1.05 [0.87 - 1.26] in the two and three trial analyses, respectively. Meta-analysis of RE-MODEL and RE-NOVATE supported the conclusions of the individual trials that dabigatran etexilate is non-inferior to enoxaparin 40 mg od, with a similar safety profile. Meta-analysis of all three trials found no significant differences between treatments in any of the end points analysed. Heterogeneity between the trials cannot be ruled out. PMID- 19132193 TI - Evaluation of enoxaparin dosing requirements in infants and children. Better dosing to achieve therapeutic levels. AB - Increasing the starting dose of enoxaparin results in the early achievement of therapeutic anti-factor Xa levels in children receiving enoxaparin which is critical for effective therapy and the reduction of venipunctures. The aim of this study was: i) to determine the enoxaparin dose required to achieve therapeutic anti-factor Xa levels in infants and children, and ii) to establish whether increasing the starting dose of enoxaparin influenced the time required to reach the therapeutic range and the number of venipunctures required for dose adjustment, and iii) the radiographic outcome of the thrombosis, where applicable. A retrospective chart review of children who received enoxaparin was carried out at the Stollery Children's Hospital, Edmonton, Alberta, Canada. Patients treated with standard-dose enoxaparin (1.5 mg/kg for children < or =3 months of age, 1.0 mg/kg for children > or =3 months of age), were compared with children who received a higher initial starting dose of enoxaparin (1.7 mg/kg for children > or =3 months of age, 1.2 mg/kg for children > or =3 months of age). Infants <3 months required an enoxaparin dose of 1.83 mg/kg, and those who received an increased initial enoxaparin dose resulted in faster attainment of therapeutic anti-factor Xa levels requiring significantly fewer venipunctures. Similarly, infants > or =3-12 months, 1-5 years, and 6-18 years, require enoxaparin 1.48 mg/kg, 1.23 mg/kg and 1.13 mg/kg, respectively, in order to achieve a therapeutic anti-factor Xa level. In conclusion, increasing the starting dose of enoxaparin may result in more rapid attainment of therapeutic range with fewer venipunctures, dose adjustments, and without an increase in adverse events. PMID- 19132194 TI - High long-term absolute risk of recurrent venous thromboembolism in patients with hereditary deficiencies of protein S, protein C or antithrombin. AB - Hereditary deficiencies of protein S, protein C and antithrombin are known risk factors for first venous thromboembolism. We assessed the absolute risk of recurrence, and the contribution of concomitant thrombophilic defects in a large cohort of families with these deficiencies. Annual incidence of recurrence was estimated in 130 deficient patients, with separate estimates for those with each of protein S, protein C, and antithrombin deficiency, and in eight non-deficient patients with prior venous thromboembolism. All patients were also tested for factor V Leiden, prothrombin G20210A, high levels of factors VIII, IX and XI, and hyperhomocysteinemia. There were 81 recurrent events among 130 deficient patients. Median follow-up was 4.6 years. Annual incidences (95% confidence interval) of recurrent venous thromboembolism were 8.4% (5.8-11.7) for protein S deficiency, 6.0% (3.9-8.7) for protein C deficiency, 10.0% (6.1-15.4) for antithrombin deficiency, and overall 7.7% (6.1-9.5). Relative risk of recurrence in patients with a spontaneous versus provoked first event was 1.5 (0.95-2.3). Cumulative recurrence rates at 1, 5 and 10 years were 15%, 38% and 53%. Relative risk of recurrence with concomitant defects was 1.4 (0.7-2.6) (1 defect) and 1.4 (0.8-2.7) (> or =2 defects). Annual incidence was 1.0% (0.03-5.5) in eight non deficient patients. Annual incidence of major bleeding in deficient patients on oral anticoagulant treatment was 0.5% (0.2-1.0). We conclude that patients with a hereditary protein S, protein C or antithrombin deficiency appear to have a high absolute risk of recurrence. This risk is increased after a first spontaneous event, and by concomitance of other thrombophilic defects. PMID- 19132195 TI - Antiphospholipid antibodies predict imminent vascular events independently from other risk factors in a prospective cohort. AB - Antiphospholipid antibodies (aPL) are associated with vascular events, but the magnitude of this risk, alone, or in combination with other atherogenic and thrombophilic risk factors, remains unclear. A prospective cohort of 415 persons was studied for arterial and venous events (AE and VE) over a median time of 7.4 years. aPL and coagulation abnormalities were measured upon beginning of the study and annually for the first four years. Within the cohort, a nested case control study was conducted to investigate the role of endothelial and inflammatory markers in predicting new vascular events. Forty-five individuals had new vascular events: 18 occurred during the first year of follow-up. The proportion of event-free survivors at eight years was 90% (95%CI = 87%, 94%) for aPL-negative and 72% (60%, 85%) for aPL-positive individuals, respectively. Predictors for new AE were previous AE (HR = 5.7 [2.7, 12.0]), diabetes (5.6 [2.4, 13.2]), aPL positivity (2.6 ([1.2, 5.9]), and age (1.04 [1.01, 1.07]). New VE were predicted by previous VE (6.1 [1.9, 19.9]), anti-beta2-glycoprotein I (abeta2GPI) positivity (5.8 [1.4, 24.1]), activated protein C resistance (APCR) (4.1 [1.1, 15.1]), and gender (3.7 [1.1, 12.9]). In the nested case-control study, similar predictors were observed for AE, while abnormal APCR (OR = 5.5 [1.1, 26.6]) and elevated von Willebrand factor (vWF) (OR = 5.0 [1.2, 19.8]) best predicted VE. We demonstrate that aPL independently predict new vascular events and discriminate between individuals with and without events in the first two years of follow-up, indicating that aPL are associated with a short-term risk of developing new and recurrent vascular events. PMID- 19132196 TI - Clopidogrel versus prasugrel in rabbits. Effects on thrombosis, haemostasis, platelet function and response variability. AB - The new P2Y(12) antagonist prasugrel produces greater inhibition of ADP-induced platelet aggregation (IPA) and reduction of thrombotic events in patients versus approved doses of clopidogrel, but increases major bleeding. We examined whether IPA level or P2Y(12) receptor occupancy (RO) could be optimized to better balance the efficacy and bleeding effects of these thienopyridines and reduce the response variability in rabbits. Rabbits were given three daily oral doses of clopidogrel (0.3-30 mg/kg/d), prasugrel (0.03-10 mg/kg/d) or vehicle (n = 6 40/group). Electrically-induced carotid artery thrombosis (AT, % thrombus weight reduction), cuticle bleeding time (BT, fold-increase over control), IPA to 20 microM ADP (% inhibition of peak light transmission) and RO (% inhibition of [(33)P]-2MeS-ADP binding to P2Y(1)-blocked platelets) were determined 2-3 hours after the last dose. ED(50) (doses for half-maximal effect, mg/kg/d) of AT, BT, IPA and RO were 1.6, 6.7, 1.9 and 1.4 for clopidogrel vs. 1.2, 1.9, 0.5 and 0.2 for prasugrel. IPA of 30-40% for both compounds produced the optimal balances of efficacy (AT: 50-60%) and BT of about 2-fold with significant RO of 70-80%. IPA of 50-60% achieved higher efficacy (AT: 60-80%), but with increased BT of five- to six-fold and >90% RO. Box-plot suggests no significant difference in the IPA and RO response variability between both compounds. Clopidogrel was 1.3-7 times less potent than prasugrel in rabbits, depending upon which biomarker was studied. The ratio of efficacy: bleeding was most favorable at a moderate IPA of 30% to 40%. Both compounds had similar IPA and RO response variability. PMID- 19132197 TI - Functional variability of platelet response to clopidogrel correlates with P2Y(12) receptor occupancy. AB - Interindividual variability of response to clopidogrel is currently a subject of much interest. We tested the hypothesis that functional variability in the platelet response to clopidogrel correlates with occupancy of the platelet P2Y(12) receptor by clopidogrel active metabolite. Healthy subjects were screened after seven days' treatment with clopidogrel 75 mg/day to select three clopidogrel-response groups (n = 12/group), defined as 'average' (40-60% inhibition of platelet aggregation [IPA]), 'low' (<10% IPA) or 'high' (>80% IPA) responders. After a two- to six-week wash-out period, subjects were randomized (double-blind) to clopidogrel 75 mg/day (n = 10/group) for 10 days, followed by clopidogrel 300 mg on day 11, or placebo (n = 2/group). IPA induced by adenosine diphosphate (ADP), and P2Y(12) receptor occupancy were measured repeatedly. The incidence of low responders was 3.7%, and low responses to clopidogrel were maintained during the randomized evaluation phase. Treatment with clopidogrel for 10 days induced a significant increase in P2Y(12) receptor occupancy in each group of responders versus placebo; receptor affinity was unchanged. This reduction correlated with IPA response (r = 0.54). The additional 300 mg dose of clopidogrel on top of 75 mg chronic treatment increased IPA and P2Y(12) receptor occupancy in all groups, but relatively more in low responders. Variability in the response to clopidogrel appears to be linked to differences in P2Y(12) receptor occupancy. An additional 300 mg dose of clopidogrel improves both IPA and P2Y(12) receptor occupancy mostly in the subset of 'low' responders. PMID- 19132199 TI - Risk of venous thromboembolism with rheumatoid arthritis. AB - Rheumatoid arthritis is not generally considered to be a risk factor for venous thromboembolism (VTE), although abnormalities of coagulation factors have been found in patients with rheumatoid arthritis. Sparse data in a few patients suggest that patients with rheumatoid arthritis may have higher rates of VTE. The purpose of this investigation was to determine if the incidences of pulmonary embolism (PE) and deep venous thrombosis (DVT) are increased in hospitalized patients with rheumatoid arthritis. The number of patients discharged from non Federal short-stay hospitals throughout the United States from 1979 through 2005 with a discharge code for rheumatoid arthritis was obtained from the National Hospital Discharge Survey (NHDS). Among hospitalized patients with rheumatoid arthritis who did not have joint surgery, 41,000 of 4,818,000 (0.85%) had PE compared with 3,366,000 of 891,055,000 (0.38%) among patients who did not have rheumatoid arthritis and who did not have operations or joint surgery (relative risk = 2.25). Deep venous thrombosis was diagnosed in 79,000 of 4,818,000 (1.64%) patients with rheumatoid arthritis and no joint operation, versus 7,681,000 of 891,055,000 (0.86%) who did not have rheumatoid arthritis or a joint operation (relative risk = 1.90). The relative risk of venous thromboembolism (PE and/or DVT) in these patients was 1.99. The data suggest that rheumatoid arthritis is a risk factor for VTE in hospitalized medical patients. A heightened awareness of the risks for VTE and a lower threshold for evaluation of patients for possible DVT or PE would be appropriate in caring for hospitalized patients with rheumatoid arthritis. PMID- 19132198 TI - Lack of association between aspirin responsiveness and seven candidate gene haplotypes in patients with symptomatic vascular disease. AB - We studied the effect of prophylactic aspirin (ASA) ingestion on platelet function in 463 patients with stroke, transient ischemic attack (TIA) or acute coronary disease (ACD), using the Platelet Function Analyzer-100 (PFA-100). We correlated ASA responsiveness with haplotypes of seven candidate genes, selected for their documented role in platelet function, namely, the genes for integrins alpha2beta1and alphaIIbbeta3 (ITGA2, ITGA2B, and ITGB3), platelet glycoproteins Ibalpha and VI (GPIBA and GP6), the purinergic receptor P2Y1 (P2RY1), and prostaglandin H synthase 1 (PTGS1 = COX1). Non-responsiveness to ASA was defined as the failure of prior ASA ingestion to prolong the PFA-100 closure time (CT) when blood was perfused through cartridges coated with collagen plus epinephrine (CEPI-CT). ASA non-responsiveness was observed in 114 of 463 patients (24.6 %), but was not associated with haplotypes of any of the seven candidate genes. There was also no association between any haplotypes and the CT when blood was perfused through cartridges coated with collagen plus ADP (CADP-CT). The ASA non responsive cohort had significantly increased whole blood platelet counts (p = 0.03) and plasma von Willebrand Factor antigen levels (p < 0.001), which likely contributes to resistance to the inhibitory effects of ASA in the PFA-100. PMID- 19132200 TI - Venous thromboembolism in critically ill patients. Observations from a randomized trial in sepsis. AB - Venous thromboembolism (VTE) is a central concern in the intensive care unit (ICU). However, little is known about both current practices for VTE prevention in the ICU and the risk for VTE in persons with severe sepsis and septic shock. XPRESS was a randomized, double-blind, placebo-controlled trial of prophylactic heparin in patients with severe sepsis and higher disease severity who were treated with drotrecogin alfa (activated) (DAA). Subjects were randomized to unfractionated heparin, low-molecular-weight heparin, or placebo during the DAA infusion period. All patients underwent ultrasonography between days 4-6 to screen for VTE. We assessed baseline utilization of VTE prophylaxis along with application of these methods after completion of the DAA infusion. The study included 1,935 subjects and, prior to enrollment approximately half were given no form of prophylaxis. By day 6, 5% of subjects developed a VTE, and the rate of VTE did not vary based on type of heparin administered. The vast majority of VTE detected by day 6 were clinically silent. Of factors analyzed, history of VTE was the only variable independently associated with development of a VTE (odds ratio, 3.66, 95% confidence interval 1.77-7.56, p = 0.005). Strikingly, patients who were initially receiving heparin prophylaxis prior to enrollment but who then had this discontinued because of randomization to placebo suffered more VTE that persons continuing on some form of heparin. Despite multiple guidelines, physicians do not uniformly prescribe VTE prophylaxis. Nonetheless, early VTE occurs even in persons given DAA. Most VTE in critically ill patients are clinically silent. PMID- 19132201 TI - Ischaemic stroke patients with heterozygous factor V Leiden present with multiple brain infarctions and widespread atherothrombotic disease. AB - Factor V Leiden (FVL) mutation is a risk factor for venous and, to a degree, arterial thrombosis. It is unknown whether and how FVL affects the manifestations of ischaemic stroke (IS). We assessed the clinical, laboratory, radiological, and prognostic characteristics in an observational study with adult IS patients having FVL. We tested 860 patients with first-ever IS for FVL and found 48 FVL positive patients. Detailed clinical, laboratory, and radiological evaluation were compared with that of 144 (1:3) gender and age matched IS patients without FVL. All patients underwent a prognostic evaluation at an average of five years follow-up. Despite the relatively young age (mean 48.5 years, range 44-54 years) of the FVL positive IS patients, peripheral arterial occlusive disease (PAOD), coronary artery disease (CAD), and previous transient cerebrovascular event occurred more frequently compared with controls. Family history of cardiovascular disease (CVD) and multiple silent brain infarctions were revealed in half of the FVL positive patients, whereas these were seldom encountered among controls. Stroke severity, long-term recovery, and recurrence rates seemed similar irrespective of FVL status. Our findings indicate that the clinical profile of IS patients with FVL associated with wider manifestation of atherothrombosis, positive family history of arterial thrombosis, and presence of multiple silent infarctions on brain images. PMID- 19132202 TI - Immature platelets in patients with acute coronary syndromes. AB - Platelets newly released from the bone marrow are RNA-containing and more haemostatically active than mature platelets. Immature platelets are reliably quantified by flow cytometry, and the immature platelet fraction (IPF) reflects platelet production and the rate of platelet turnover. It was the objective of this study to evaluate the presence of immature platelets in healthy subjects, patients with stable coronary artery disease (CAD) and patients with acute coronary syndromes. Flow cytometric determination of immature platelets was performed with an automated analyzer (Sysmex XE-2100) using RNA fluorescent dyes. IPF was determined in 420 individuals: 22 healthy subjects, 39 patients with stable CAD, 182 patients with unstable angina/non-ST-segment elevation myocardial infarction (non-STEMI) and 177 patients with acute STEMI. The geometric mean [95% confidence interval] of IPF was 2.51 [2.04-3.10] in healthy subjects, 2.87 [2.45 3.36] in CAD patients, 2.93 [2.72-3.15] in the non-STEMI/unstable angina group and 3.71 [3.45-3.99] in patients with STEMI (ANOVA: p < 0.0001). This difference remained significant after adjusting for baseline characteristics (p = 0.0003). In active smokers, IPF was 18% higher than in non-smoking individuals (p = 0.007), and IPF was 16% higher in diabetics compared with non-diabetics (p = 0.060). In conclusion, the fraction of immature platelets is increased in acute coronary syndromes, especially in the acute phase of STEMI. Immature platelets with an increased haemostatic potential may contribute to coronary thrombus formation. PMID- 19132203 TI - No effect of lipid lowering on platelet activity in patients with coronary artery disease and type 2 diabetes or impaired glucose tolerance. AB - In addition to lowering cholesterol, statins may reduce platelet activity and exert beneficial non-lipid (pleiotropic) effects. We evaluated the effects of two different simvastatin based treatment regimens on platelet reactivity in patients with dysglycemia and coronary artery disease (CAD). Thirty-two patients with type 2 diabetes or impaired glucose tolerance and stable CAD received six weeks of double-blind treatment with simvastatin 80 mg daily (S80; n = 16) or ezetimibe 10 mg and simvastatin 10 mg daily (E10/S10; n = 16). Total and low-density lipoprotein (LDL) cholesterol, and high sensitivity C-reactive protein (CRP) decreased similarly in the two groups. LDL (mM) decreased from 3.2 +/- 0.6 to 1.7 +/- 0.7 with E10/S10 and from 3.0 +/- 1.0 to 1.4 +/- 0.5 with S80 treatment. Platelet function was evaluated by whole blood flow cytometry and turbidimetric aggregometry with agonist stimulation ex vivo before and after treatment. Neither treatment affected basal or adenosine diphosphate (ADP)- or thrombin-induced platelet P-selectin expression, or fibrinogen binding, or platelet-leukocyte aggregation. Similarly, neither treatment affected ADP-induced platelet aggregation. In conclusion, lipid lowering treatment with high dose simvastatin or low dose simvastatin plus ezetimibe did not exert any substantial inhibitory effects on the basal or agonist-stimulated activity of circulating platelets in patients with stable CAD and type 2 diabetes or impaired glucose tolerance. PMID- 19132204 TI - The technique of measuring thrombin generation with fluorogenic substrates: 3. The effects of sample dilution. AB - Assessing the clotting function inevitably brings about dilution of plasma. With the existing techniques of thrombin generation (TG) measurement, dilution ranges from 2:3 to 1:8. However, the possibility that dilution alters procoagulant and anticoagulant pathways differently has not been examined. We investigated the effects of dilution on the thrombin generation process and found that the anticoagulant pathways are far more affected by dilution than the procoagulant pathways. That is, when prothrombin and antithrombin concentrations are kept constant, dilution of plasma does not significantly affect tissue factor (TF) driven thrombin generation. We demonstrate that dilution of plasma slows down the inhibitory activity of tissue factor pathway inhibitor (TFPI) to a greater extent when compared with the down regulation by diluting procoagulant factors. Dilution of plasma has also a negative effect on the participation of the antihaemophiliac factors VIII and IX in TG driven by contact activation or low TF concentration. We also investigated the effect of dilution on the participation of the anticoagulant system that consists of thrombomodulin, protein C and protein S (APC system). We found that plasma dilution causes a loss of sensitivity towards TM and APC. Furthermore, at high dilutions (> 1:12) a second wave of prothrombinase-activity was observed that could be attributed to the suppression of protein S-dependent inhibition. In conclusion, the mechanism of TG is profoundly disturbed by plasma dilution. As a consequence, the less a plasma sample is diluted, the better a TG experiment represents the physiological process. PMID- 19132205 TI - The technique of measuring thrombin generation with fluorescent substrates: 4. The H-transform, a mathematical procedure to obtain thrombin concentrations without external calibration. AB - In fluorogenic thrombin generation (TG) experiments, thrombin concentrations cannot be easily calculated from the rate of the fluorescent signal increase, because the calibration coefficient increases during the experiment, due to substrate consumption and quenching of the fluorescent signal by the product. Continuous, external calibration via an in a parallel sample therefore was hitherto required for an accurate calculation of the TG curve. A technique is presented that allows mathematical transformation of experimental fluorescence intensities into "ideal" data, i.e. in the data that would have been obtained if substrate consumption and quenching by the product would not play a role. The method applies to fluorescence intensities up to 90% of the maximal fluorescent signal corresponding to total substrate conversion and thereby covers the entire region of interest encountered in practice. The first derivative of the transformed signal can then be converted into thrombin concentrations via a conventional, fixed calibration factor. This calibration factor can be obtained from a separate experiment but also by measuring the amidolytic activity of the alpha(2)macroglobulin-thrombin complex present in the reaction mixture ("serum") after thrombin generation is over. This method halves the amount of sample required per experiment. PMID- 19132206 TI - Are laboratories following published recommendations for lupus anticoagulant testing? An international evaluation of practices. AB - Laboratory tests for lupus anticoagulants (LA) are commonly performed to evaluate thrombosis or suspected phospholipid antibody syndromes. To determine current LA testing practices, and if they conform to published recommendations, two questionnaires were distributed to clinical laboratory members of the North American Specialized Coagulation Laboratory Association (NASCOLA) and the ECAT Foundation (ECAT). The first and second questionnaires were completed by 113 and 96 laboratories, respectively. Commonly performed LA tests included the dilute Russell's viper venom time, LA sensitive activated partial thromboplastin time and hexagonal phospholipid test. Although some laboratories did single LA tests if requested, the majority complied with published recommendations: to use platelet poor plasma for LA tests; to use two or more screening tests, representing different assay principles, and one assay having a low phospholipid concentration to exclude LA; to confirm LA phospholipid dependency by the method giving an abnormal LA screen; to document the inhibitor activity on pooled normal plasma; and not to use phospholipid antibodies to confirm LA. A minority (<35%) followed the recommendations to exclude factor deficiencies and factor inhibitors as the cause of an abnormal LA test. After participating, 32% of laboratories had changed practices and 20% indicated that they would be changing practices. While most laboratories generally follow published guidelines for LA testing, few follow recommendations to evaluate for other coagulation abnormalities. Questionnaires may be helpful quality initiatives to improve compliance with laboratory testing guidelines and recommendations. PMID- 19132207 TI - Laboratory detection of the antiphospholipid syndrome via calibrated automated thrombography. AB - Lupus anticoagulants (LAC) consist of antiphospholipid antibodies, detected via their anticoagulant properties in vitro. Strong LAC relate to thromboembolic events, a hallmark of the antiphospholipid syndrome. We have analyzed whether detection of this syndrome would benefit from thrombin generation measurements. Therefore, calibrated automated thrombography was done in normal plasma (n = 30) and LAC patient plasma (n = 48 non-anticoagulated, n = 12 on oral anticoagulants), diluted 1:1 with a normal plasma pool. The anti-beta2 glycoprotein I monoclonal antibody 23H9, with known LAC properties, delayed the lag time and reduced the peak height during thrombin generation induction in normal plasma dose-dependently (0-150 microg/ml). At variance, LAC patient 1:1 plasma mixtures manifested variable lag time prolongations and/or peak height reductions. Coupling these two most informative thrombin generation parameters in a peak height/lag time ratio, and upon normalization versus the normal plasma pool, this ratio distributed normally and was reduced in the plasma mixtures, for 59/60 known LAC plasmas. The normalized peak height/lag time ratio correlated well with the normalized dilute prothrombin time, diluted Russell's viper venom time and silica clotting time, measured in 1:1 plasma mixtures (correlation coefficients 0.59-0.72). The anticoagulant effects of activated protein C (0-7.5 nM) or 23H9 (0-150 microg/ml), spiked in the 1:1 LAC plasma mixtures were reduced for the majority of patients, compatible with functional competition between patient LAC and activated protein C and LAC and 23H9, respectively. Hence, the normalized thrombin generation-derived peak height/lag time ratio identifies LAC in plasma with high sensitivity in a single assay, irrespective of the patient's treatment with oral anticoagulants. PMID- 19132208 TI - Validity and clinical utility of the simplified Wells rule for assessing clinical probability for the exclusion of pulmonary embolism. AB - The recently introduced simplified Wells rule for the exclusion of pulmonary embolism (PE) assigns only one point to the seven variables of the original Wells rule. This study was performed to independently validate the simplified Wells rule for the exclusion of PE. We retrospectively calculated the prevalence of PE in the "unlikely" probability categories of the original Wells (cut-off < or =4) and the simplified Wells rule (cut-off < or =1) in 922 consecutive patients with clinically suspected PE from a multicenter cohort study. We compared the three month incidence of venous thromboembolism (VTE) in patients with an unlikely probability and a normal D-dimer test using both scores, and the proportion of patients with this combination (clinical utility). The proportion of patients categorized as PE "unlikely" was similar using the original (78%) and the simplified (70%) Wells rule. The prevalence of PE was 13% (95% confidence interval [CI], 11-16%) and 12% (95%CI, 9.7-15%) for the original Wells and simplified Wells "unlikely" categories, respectively. None of the patients with PE "unlikely" and a normal D-dimer test experienced VTE during three-month follow up. The proportions of patients in whom further tests could safely be withheld based on PE "unlikely" and a normal D-dimer test was 28% (95%CI, 25-31%) using the original and 26% (95%CI, 24-29%) using the simplified Wells rule. In this external retrospective validation study, the simplified Wells rule appeared to be safe and clinically useful, although prospective validation remains necessary. Simplification of the Wells rule may enhance the applicability. PMID- 19132209 TI - Drug-eluting balloon: very short-term exposure and overlapping. AB - Paclitaxel balloon coating has shown promising effects in inhibiting restenosis in initial clinical trials. The aim of the present study was to evaluate the influence of two critical features of drug-eluting balloon (DEB) application - inflation time and increased dose due to overlapping balloons. Fifty-six stainless steel stents were implanted in the left anterior descending and circumflex coronary arteries of 28 domestic pigs using a 1.2:1.0 overstretch ratio. Stents were mounted on conventional uncoated and paclitaxel-coated angioplasty balloon catheters. The animals were randomized to five different treatments with a range of short (10 seconds [s] inflation using 1 DEB) to extended (2x60 s inflation using 2 DEB) intima contact time. After 28 days, quantitative angiography and histomorphometry of the stented arteries was performed on a total of 23 pigs. Paclitaxel balloon coating led to a marked reduction of parameters characterizing in-stent stenosis: Late lumen loss was 1.37 +/- 0.49 mm for uncoated balloons, 0.23 +/- 0.42 mm for one coated balloon 60 s inflation time, 0.37 +/- 0.28 mm for 10 s inflation time and 0.30 +/- 0.19 mm for the vessel segment treated by two coated balloons with 60 s inflation each. Neointimal areas were 4.26 +/- 1.18, 1.68 +/- 0.23, 1.83 +/- 0.40 and 1.67 +/- 0.46 mm(2), respectively (p = 0.001 versus control, p > 0.05 between paclitaxel-treated groups). Despite the marked reduction of neointimal proliferation, endothelialization of stent struts was present in all samples. DEB were found to effectively reduce neointimal proliferation regardless of inflation time and dose within the tested range. No adverse reactions were seen as dose was increased to more than three times the clinically tested dose. PMID- 19132210 TI - Multiple electrode aggregometry for ex-vivo detection of the anti-platelet effect of non-opioid analgesic drugs. PMID- 19132211 TI - Low absolute risk of venous and arterial thrombosis in hyperhomocysteinaemia - a prospective family cohort study in asymptomatic subjects. PMID- 19132212 TI - Haplotypic or genotypic combinations of three protein Z polymorphisms influence protein Z plasma level. PMID- 19132213 TI - Plasma Gas6 levels and coronary artery disease. PMID- 19132214 TI - Bleeding risks with combination of oral anticoagulation plus antiplatelet therapy: is clopidogrel any safer than aspirin when combined with warfarin? PMID- 19132215 TI - Difference between sepsis and trauma diagnosed as sensitive diagnostic criteria for disseminated intravascular coagulation. PMID- 19132216 TI - Leptin, obesity and platelet responsiveness: another piece in the puzzle. PMID- 19132217 TI - Predicting thrombotic events: creating a complex approach for a complex condition. PMID- 19132218 TI - Plasminogen activator inhibitor-1 (PAI-1): a molecule at the crossroads to cell survival or cell death. PMID- 19132219 TI - Regulation of plasminogen activator inhibitor type 1 gene expression by inflammatory mediators and statins. AB - Elevated plasma concentrations of plasminogen activator inhibitor type 1 (PAI-1), also named serpin E1, are encountered in patients with thrombophilia, atherosclerosis, septicemia and the metabolic syndrome and may be associated with an increased risk of complications. Expression of PAI-1 is increased by inflammatory stimuli and decreased by statins, drugs widely used in patients with cardiovascular disease. Increased expression of PAI-1 by inflammatory stimuli is mediated by a large variety of signal transduction pathways, which include the NF kappaB and MAP kinase pathways. The downregulating effect of statins on PAI-1 expression is dependent on the inhibition of Rho family proteins and may involve an activation of PI-3 kinase/Akt signaling pathways. In this review we summarize the findings on the effect of inflammation and statins on PAI-1 expression. PMID- 19132221 TI - The 'PAI-1 paradox' in vascular remodeling. AB - Vascular remodelling is a complex phenomenon associated with restructuring of the vessel wall as a consequence of disruption of vascular homeostasis. Alterations of the vascular wall have been linked to a variety of cardiovascular disorders including atherosclerosis, vascular injury and pulmonary hypertension. Plasminogen activator inhibitor-1 (PAI-1) is a member of the serpin (serine proteinase inhibitor) family and acts as an important inhibitor of fibrinolysis by interfering with the plasminogen system. In addition to its anti-fibrinolytic effects, PAI-1 appears to modulate cellular responses linked to vascular remodelling. Since PAI-1 levels have been shown to be altered in various disorders associated with vascular remodelling of the systemic and pulmonary vascular bed, this serpin may play a pivotal role in the pathogenesis of these diseases. PMID- 19132220 TI - Integration of non-SMAD and SMAD signaling in TGF-beta1-induced plasminogen activator inhibitor type-1 gene expression in vascular smooth muscle cells. AB - Overexpression of plasminogen activator inhibitor-1 (SERPINE1, PAI-1), the major physiological inhibitor of pericellular plasmin generation, is a significant causative factor in the progression of vascular disorders (e.g. arteriosclerosis, thrombosis, perivascular fibrosis) as well as a biomarker and a predictor of cardiovascular-disease associated mortality. PAI-1 is a temporal/spatial regulator of pericellular proteolysis and ECM accumulation impacting, thereby, vascular remodeling, smooth muscle cell migration, proliferation and apoptosis. Within the specific context of TGF-beta1-initiated vascular fibrosis and neointima formation, PAI-1 is a member of the most prominently expressed subset of TGF-beta1-induced transcripts. Recent findings implicate EGFR/pp60c-src- >MEK/ERK1/2 and Rho/ROCK-->SMAD2/3 signaling in TGF-beta1-stimulated PAI-1 expression in vascular smooth muscle cells. The EGFR is a direct upstream regulator of MEK/ERK1/2 while Rho/ROCK modulate both the duration of SMAD2/3 phosphorylation and nuclear accumulation. E-box motifs (CACGTG) in the PE1/PE2 promoter regions of the human PAI-1 gene, moreover, are platforms for a MAP kinase-directed USF subtype switch (USF-1-->USF-2) in response to growth factor addition suggesting that the EGFR-->MEK/ERK axis impacts PAI-1 expression, at least partly, through USF-dependent transcriptional controls. This paper reviews recent data suggesting the essential cooperativity among the EGFR-->MAP kinase cascade, the Rho/ROCK pathway and SMADs in TGF-beta1-initiated PAI-1 expression. The continued clarification of mechanistic controls on PAI-1 transcription may lead to new targeted therapies and clinically-relevant options for the treatment of vascular diseases in which PAI-1 dysregulation is a major underlying pathogenic feature. PMID- 19132222 TI - Metabolic, hormonal and environmental regulation of plasminogen activator inhibitor-1 (PAI-1) expression: lessons from the liver. AB - Plasminogen activator inhibitor-1 (PAI-1) controls the regulation of the fibrinolytic system in blood by inhibiting both urokinase-type and tissue-type plasminogen activators. Enhanced levels of PAI-1 are found in patients with type 2 diabetes mellitus which is associated with a dysbalance in glucose and lipid homeostasis. Especially a defective insulin response in the liver contributes to the development of hyperglycemia, dyslipidemia and peripheral insulin resistance and may contribute to hepatic over-expression of PAI-1 in diabetes type 2. Furthermore, a substantial upregulation of PAI-1 expression has also been shown in a variety of liver injury models. Thus, the liver appears to be not only a major site of PAI-1 synthesis in response to hormonal changes, but also in response to a variety of other pathological events. PAI-1 expression in liver largely depends on activation of signalling pathways and transcriptional regulators which may be the basis for a new level of cross-talk between different signalling pathways and thus may represent attractive therapeutic candidates. This article will primarily focus on the regulation of PAI-1 expression in liver cells and discuss potential cross-talks between metabolic, hormonal and environmental signals. PMID- 19132223 TI - Transcriptional regulation of the plasminogen activator inhibitor type 1--with an emphasis on negative regulation. AB - By inhibiting plasminogen activators uPA and tPA, inducing uPA-uPAR internalization and interfering with the interaction between extracellular matrix protein vitronectin and alphavbeta3 integrin, plasminogen activator inhibitor type 1 (PAI-1) is active in the regulation of various biological processes involving extracellular proteolysis and tissue remodeling. PAI-1 is expressed in many cell types under the control of a variety of signals, depending on cell type. The most prominent and important of these signals are TGFbeta, hypoxia and insulin. Although the signaling pathways were largely elucidated, recent investigations have revealed more complicated aspects. The pathways interact at the level of both transcription factors and regulatory elements on the promoter. Furthermore, the engagement of negative factors in these pathways has been shown to be important, adding complexity and versatility to PAI-1 gene regulation. PMID- 19132224 TI - Anti-apoptotic roles of plasminogen activator inhibitor-1 as a neurotrophic factor in the central nervous system. AB - Plasminogen activator inhibitor-1 (PAI-1), a member of the serpin gene family, is the primary inhibitor of urokinase-type and tissue-type PAs. PAI-1 plays an important role in the process of peripheral tissue remodeling and fibrinolysis through the regulation of PA activity. This serpin is also produced in brain tissues and may regulate the neural protease sequence in the central nervous system (CNS), as it does in peripheral tissues. In fact, PAI-1 mRNA is up regulated in mouse brain after stroke. The serpin activity of PAI-1 helps to prevent tissue-type PA-induced neuron death. However, we have previously found that PAI-1 has a novel biological function in the CNS: the contribution to survival of neurites on neurons. In neuronally differentiated rat pheochromocytoma (PC-12) cells, a deficiency of PAI-1 in vitro caused a significant reduction in Bcl-2 and Bcl-X(L) mRNAs and an increase in Bcl-X(S) and Bax mRNAs. The change in the balance between mRNA expressions of the anti- and pro-apoptotic Bcl-2 family proteins promoted the apoptotic sequence: caspase-3 activation, cytochrome c release from mitochondria and DNA fragmentation. Our results indicate that PAI-1 has an anti-apoptotic role in neurons. PAI-1 prevented the disintegration of the formed neuronal networks by maintaining or promoting neuroprotective signaling through the MAPK/ERK pathway, suggesting that the neuroprotective effect of PAI-1 is independent of its action as a protease inhibitor. This review discusses the neuroprotective effects of PAI-1 in vitro, together with the relevant data from other laboratories. Special emphasis is placed on its action on PC-12 cells. PMID- 19132225 TI - Rac-1 promotes pulmonary artery smooth muscle cell proliferation by upregulation of plasminogen activator inhibitor-1: role of NFkappaB-dependent hypoxia inducible factor-1alpha transcription. AB - Pulmonary vascular remodeling is commonly associated with pulmonary hypertension and is characterized by media thickening and disordered cellular proliferation, often accompanied by fibrin deposition and thrombosis in situ. However, the signaling pathways linking these different processes are not well understood. Since the GTPase Rac-1 has been suggested to act as a signaling relay in various cell types we investigated whether Rac-1 could be the link between thrombin signaling, plasminogen activator inhibitor-1 (PAI-1), which inhibits fibrinolysis and promotes fibrin deposition, and proliferation of pulmonary artery smooth muscle cells (PASMC). Exposure to thrombin enhanced the levels of Rac-1 protein and increased PAI-1 mRNA and protein expression in dependence of the thrombin receptor PAR-1. Expression of dominant-negative Rac-1 (RacT17N) prevented thrombin-induced PAI-1 expression whereas constitutively active RacG12V enhanced PAI-1 levels. In the presence of RacT17N thrombin-induced PAI-1 promoter activity was abrogated whereas RacG12V increased PAI-1 promoter activity, and this response was essentially dependent on the transcription factor hypoxia-inducible factor-1 (HIF-1). Subsequently, RacG12V not only increased HIF transcriptional activity but also HIF-1alpha protein and mRNA levels, whereas RacT17N prevented these responses elicited by thrombin. In line, RacG12V enhanced HIF-1alpha promoter activity, and this response was dependent on nuclear factor-kappaB (NFkappaB) binding to the HIF-1alpha promoter. Finally, upregulation of PAI-1 by Rac-1 and HIF-1 was essential for thrombin-stimulated proliferation of PASMC. These findings indicate that Rac-1 is an important mediator of thrombin signaling and may contribute to pulmonary vascular remodeling via HIF-1-dependent upregulation of PAI-1 leading to enhanced proliferation of PASMC. PMID- 19132227 TI - The effect of plasminogen activator inhibitor type 1 on apoptosis. AB - Plasminogen activator inhibitor type-1 (PAI-1), an inhibitor of plasminogen activators, inhibits formation of plasmin and plasmin-mediated proteolysis. Apoptosis, or programmed cell death, is a potentially important phenomenon in mediating overall cell death. This review focuses on the influence of PAI-1 on apoptosis. Greater expression of PAI-1 has been associated with increased survival of cells and resistance to apoptosis. PAI-1 appears to influence apoptosis by decreasing cell adhesion (anoikis) as well as its effect on intracellular signaling. Mechanisms by which PAI-1 may render a cell resistant to apoptosis include its ability to inhibit generation of plasmin, its ability to inhibit caspase 3, and its ability to inhibit cell adhesion mediated by vitronectin. Inhibition of caspase 3 by PAI-1 may divert intracellular signalling from induction of apoptosis to induction of proliferation. PMID- 19132226 TI - Plasminogen activator inhibitor-1: the double-edged sword in apoptosis. AB - Plasminogen activator inhibitor type-1 (PAI-1) is a multi-functional protein. It is a fast-acting inhibitor of plasminogen activators; urokinase-plasminogen activator and tissue type plasminogen activator, and also plays an important role in regulating cell proliferation, adhesion, migration, and signal transduction pathways. These biological events are important processes during angiogenesis and restenosis. PAI-1 has been shown to regulate proliferation, migration, and apoptosis of vascular smooth muscle cells and endothelial cells. The ability of PAI-1 to regulate cellular proliferation and migration has been attributed to its ability to control plasmin production, modify signaling pathways, and its inherent multifactorial ability to bind to vitronectin and lipoprotein receptor related protein. However, the mechanism by which PAI-1 regulates the apoptotic pathway is not well understood. Evidence from the literature suggests that PAI-1 or its deficiency alters key signalling pathways, such as the PI3-k/Akt and the Jak/STAT pathways, and is involved in maintaining endothelial cell integrity thereby regulating cell death. Other investigators have demonstrated that PAI-1 directly binds to caspases as a mechanism of PAI-1-mediated cellular apoptosis. Moreover, results from studies assessing the role of PAI-1 in apoptosis have suggested that PAI-1 can exert pathogenic or protective effects, which may be related to the disease model or type of injury employed. PMID- 19132228 TI - Regulation of programmed cell death by plasminogen activator inhibitor type 1 (PAI-1). AB - Elevated levels of plasminogen activator inhibitor-1 (PAI-1) are associated with poor prognosis in cancer. An explanation to the elevated levels of PAI-1 could be a protective response to the increased proteolytic activity, caused by elevated levels of urokinase-type plasminogen activator (uPA) observed in tumours; however, several lines of evidence suggest that PAI-1 may contribute directly to the pathology of the disease. PAI-1 has been reported to have an effect on most of the basic cellular processes including cell adhesion, cell migration, cell invasion, and cell proliferation and increasing numbers of reports suggest that PAI-1 also can regulate programmed cell death (PCD) in cancer cells and normal cells. A number of reports suggest that PAI-1 can inhibit PCD through its pro adhesive/anti-proteolytic property whereas other reports suggest that PAI-1 induces PCD through its anti-adhesive property. Furthermore, it has been suggested that PAI-1 can either induce or inhibit PCD though activation of cell signalling pathways. This review will focus on the regulation of programmed cell death by PAI-1 in both normal cells and cancer cells. PMID- 19132229 TI - Bleeding complications in patients on anticoagulants who would have been disqualified for clinical trials. PMID- 19132230 TI - Pharmacogenetic differences between warfarin, acenocoumarol and phenprocoumon. AB - Coumarin oral anticoagulant drugs have proven to be effective for the prevention of thromboembolic events. World-wide, warfarin is the most prescribed drug. In Europe, acenocoumarol and phenprocoumon are also administered. Yet it has been proven that variant alleles of the VKORC1 and CYP2C9 genotypes influence the pharmacokinetics and pharmacodynamics of these drugs. The combination of these two variant genotypes is a major cause of the inter-individual differences in coumarin anticoagulant drug dosage. Individuals who test positive for both variant genotypes are at increased risk of major bleeding. The impact of the CYP2C9 and VKORC1 genotype is most significant during the initial period of coumarin anticoagulant therapy. The effect of VKORC1 allelic variants is relatively similar for all three VKAs. The CYP2C9 polymorphism is associated with delayed stabilisation for coumarin anticoagulants. The effects of CYP2C9 polymorphisms on the pharmacokinetics and anticoagulant response are least pronounced in the case of phenprocoumon. In the long term, patients using phenprocoumon have more often international normalised ratio (INR) values in the therapeutic range, requiring fewer monitoring visits. This leads us to conclude that in the absence of pharmacogenetic testing, phenprocoumon seems preferable for use in long-term therapeutic anticoagulation. Pharmacogenetic testing before initiating coumarin oral anticoagulants may add to the safety of all coumarin anticoagulants especially in the elderly receiving multiple drugs. PMID- 19132231 TI - Multimerin 1 binds factor V and activated factor V with high affinity and inhibits thrombin generation. AB - Multimerin 1 (MMRN1) is a polymeric, factor V (FV) binding protein that is stored in platelet and endothelial cell secretion granules but is undetectable in normal plasma. In human platelet alpha-granules, FV is stored complexed to MMRN1, predominantly by noncovalent binding interactions. The FV binding site for MMRN1 is located in the light chain, where it overlaps the C1 and C2 domain membrane binding sites essential for activated FV (FVa) procoagulant function. Surface plasmon resonance (SPR), circular dichroism (CD) and thrombin generation assays were used to study the binding of FV and FVa to MMRN1, and the functional consequences. FV and FVa bound MMRN1 with high affinities (K(D): 2 and 7 nM, respectively). FV dissociated more slowly from MMRN1 than FVa in SPR experiments, and CD analyses suggested greater conformational changes in mixtures of FV and MMRN1 than in mixtures of FVa and MMRN1. SPR analyses indicated that soluble phosphatidylserine (1,2-Dicaproylsn-glycero-3-phospho-L-serine) competitively inhibited both FV-MMRN1 and FVa-MMRN1 binding. Furthermore, exogenous MMRN1 delayed and reduced thrombin generation by plasma and platelets, and it reduced thrombin generation by preformed FVa. Exogenous MMRN1 also delayed FV activation, triggered by adding tissue factor to plasma, or by adding purified thrombin or factor Xa to purified FV. The high affinity binding of FV to MMRN1 may facilitate the costorage of the two proteins in platelet alpha-granules. As a consequence, MMRN1 release during platelet activation may limit platelet dependent thrombin generation in vivo. PMID- 19132232 TI - Modulation of tissue factor and tissue factor pathway inhibitor-1 by neutrophil proteases. AB - During systemic inflammation, neutrophil activation is accompanied by endothelial cell damage and hypercoagulability. Activated neutrophils release serine proteases that participate in tissue injury. We sought to investigate the effects of neutrophil proteases on proinflammatory and procoagulant changes in endothelial cells. The effects of elastase (HNE), cathepsin G (CG), and proteinase 3 (PR3) on expression of tissue factor (TF) and tissue factor pathway inhibitor-1 (TFPI) were examined in human umbilical vein endothelial cells. Flow cytometry demonstrated that these proteases proteolytically degraded endothelial cell-bound TFPI. TFPI mRNA expression was reduced by HNE and CG. PR3, but not HNE or CG, increased surface expression of TF and TF mRNA. Yet, increased TF expression did not enhance TF activity suggesting induction of encrypted TF. Using antibodies and siRNA to inhibit and silence PAR-1 and PAR-2, we observed that PR3 upregulation of TF is at least in part mediated by PAR-1. Although CG and HNE cleaved PAR-1, antibody reactivity to the PAR-1 hirudin-like sequence demonstrated inactivating cleavage, accounting for the selective ability of PR3 to induce PAR-1-mediated procoagulant effects. This was supported by induction of p42/44 MAPK by PR3. In conclusion, PR3 degradation of TFPI increases the procoagulant activity of endothelial cells. Release of PR3 after neutrophil activation may represent an important step in neutrophil-mediated vascular injury. PMID- 19132233 TI - Effect of oral antiplatelet agents on major bleeding in users of coumarins. AB - Treatment with vitamin K antagonists (coumarins) is associated with an increased risk of bleeding. In order to elucidate the bleeding risk of users of antiplatelet drugs among users of coumarins, we assessed the odds ratio of major bleeding associated with use of antiplatelet drugs in users of the coumarins acenocoumarol and phenprocoumon. We used data from a Dutch record linkage system, including pharmacy and linked hospitalization records for approximately two million subjects, to conduct a nested case control study in a cohort of new users of coumarins. Cases were patients who were hospitalized with a primary diagnosis of major bleeding while taking coumarin and were matched with up to four control subjects. Conditional logistic regression analysis was used to determine ORs and 95% confidence intervals (CI). We identified 1848 case patients who were matched to 5818 controls. Users of clopidogrel or aspirin showed a significantly increased risk of hospitalization because of major bleeding (OR 2.9, 95% CI 1.2 6.9 and OR 1.6, 95% CI 1.3-1.9, respectively), whereas users of dipyridamole and combinations of antiplatelet drugs showed a strong trend (OR 1.5, 95% CI 1.0-2.3 and OR 1.8, 95 % CI 1.0-3.3, respectively). In all cases, the risks were greater for upper gastrointestinal bleedings than for other bleedings. In conclusion, the use of any antiplatelet drug increases the risk of hospitalization for major bleeding among users of coumarins. Concurrent use of clopidogrel or dipyridamole and coumarins is probably not safer than concurrent use of aspirin and coumarins. PMID- 19132234 TI - Risks and benefits of transcatheter thrombolytic therapy in patients with splanchnic venous thrombosis. AB - Transcatheter local thrombolytic therapy in patients with acute, extended splanchnic venous thrombosis is controversial. Here we present our single-center experience with transcatheter thrombolytic therapy in these patients. All consecutive patients (n = 12) with acute, extended splanchnic venous thrombosis who underwent transcatheter thrombolytic therapy in our hospital, were included in this study. Thrombolytic therapy was successful for three thrombotic events and partially successful for four thrombotic events. Two patients developed minor procedure-related bleeding (17%). Six patients (50%) developed major procedure related bleeding, with a fatal outcome in two. Transcatheter thrombolytic therapy in patients with acute, extended splanchnic vein thrombosis is found to be associated with a high rate of procedure-related bleeding. Therefore, thrombolysis should be reserved for patients in whom the venous flow cannot be restored by using conventional anticoagulant therapy or stent placement across the thrombosed vessel segment. PMID- 19132235 TI - Self-management of oral anticoagulation reduces major outcomes in the elderly. A randomized controlled trial. AB - Although many patients with long-term oral anticoagulation (OAC) can manage their medication safely and reliably themselves, no study on elderly patients has as yet assessed the safety and efficacy of OAC self-management with major thromboembolic and haemorrhagic complications as primary outcomes. In this multi centre trial, patients aged 60 years or more were randomised into a self management (SMG) (N = 99) or routine care group (RCG) (N = 96). The primary outcome was the combined endpoint of all thromboembolic events requiring hospitalisation and all major bleeding complications. Mean follow-up was 2.9 +/- 1.2 and 3.0 +/- 1.1 years in the SMG and RCG, respectively. In intention-to-treat analysis, 12 patients in the SMG versus 22 patients in the RCG reached a primary endpoint (hazard ratio [HR]: 0.50; 95% confidence interval [CI]: 0.25 to 1.00; p = 0.049). The post-hoc analysis on OAC treatment sensitivity supported the benefit of self-management (HR 0.27; 95% CI: 0.13 to 0.71; p = 0.006). Fifteen patients in the SMG died during the study, but none of the deaths was directly associated with anticoagulation therapy. In the RCG, 11 patients died; of those, three deaths were directly associated with anticoagulation therapy and there was one death of unknown cause. During follow-up, the quality of OAC control was significantly better in the SMG than in the RCG. In elderly patients, long-term self-management of oral anticoagulation is superior for the prevention of major thromboembolic and bleeding complications and for the quality of oral anticoagulation control compared to routine care for a mean follow-up period of three years. PMID- 19132237 TI - Peculiar whole blood rotation thromboelastometry (Rotem) profile in 40 sideropenic anaemia patients. AB - The ROtation ThromboElastoMetry analyser (ROTEM, Pentapharm, Munich, Germany) is useful for studying whole blood (WB) clot formation and lysis. Reduction of haematocrit (HCT) has been reported to influence traditional thromboelastography parameters without compromising "in vitro" blood coagulation. We performed this case-control study to evaluate ROTEM profiles in sideropenic anaemia patients with different degrees of reduction of HCT levels. Forty consecutively referred patients with sideropenic anaemia were enrolled. A group of 40 healthy age and gender matched subjects acted as a control. The influence of HCT on ROTEM was assessed in the study population and in a model of artificially reconstituted blood with modified HCT values. Cases presented significantly increased levels of maximum clot firmness (MCF) as compared to controls (p < 0.001) mimicking a sort of "hypercoagulable profile". However, thrombin generation tests failed to detect an increase in thrombin generation in cases as compared to controls. A statistically significant inverse linear correlation between HCT and MCF (p < 0.0001) was found. In addition, ROTEM profiles following "in vitro" manipulation of HCT confirmed the inverse linear correlation between HCT and MCF found in the study population. In conclusion, the increased clot firmness found by ROTEM in anaemic patients is likely to be related to the method in itself rather than representing a marker of hypercoagulability "in vivo". Since ROTEM is widely used by anaesthesiologists when deciding the optimisation of products supplementation during surgery, attention should be paid in the case of anaemic patients taking depending on the peculiar thrombo-elastography profile found. PMID- 19132236 TI - Clinical course and outcome of disseminated intravascular coagulation diagnosed by Japanese Association for Acute Medicine criteria. Comparison between sepsis and trauma. AB - The Japanese Association for Acute Medicine (JAAM) disseminated intravascular coagulation (DIC) study group recently announced new diagnostic criteria for DIC. These criteria have been prospectively validated and demonstrated to progress to overt DIC as defined by the International Society on Thrombosis and Haemostasis (ISTH). Although an underlying condition is essential for the development of DIC, it has never been clarified if patients with different underlying disorders have a similar course. Among 329 patients with DIC diagnosed by the JAAM criteria, those with underlying sepsis (n = 98) or trauma (n = 95) were compared. The 28 day mortality rate was significantly higher in sepsis patients than trauma patients (34.7% vs. 10.5%, p < 0.0001). Within three days of fulfilling the JAAM criteria, sepsis patients had a lower platelet count, higher prothrombin time ratio, higher systemic inflammatory response syndrome score, and higher Sequential Organ Failure Assessment score compared with trauma patients. On day 3, a significantly higher percentage of trauma patients than sepsis patients showed improvement of DIC (64.2% vs. 30.6%, p < 0.001). These differences were mainly due to patients with lower JAAM DIC scores. More than 50% of the JAAM DIC patients with sepsis who died within 28 days could not be detected by ISTH DIC criteria during the initial three days. In contrast, most trauma patients who died within 28 days had DIC simultaneously diagnosed by JAAM and ISTH criteria, except for those with brain death. These findings suggest that coagulation abnormalities, organ dysfunction, and the outcome of JAAM DIC differ between patients with sepsis and trauma. PMID- 19132239 TI - Absence of leptin resistance in platelets from morbidly obese individuals may contribute to the increased thrombosis risk in obesity. AB - Clinical studies have shown that elevated leptin levels are an independent cardiovascular risk factor. However, little is known about the existence of platelet resistance to leptin in the setting of obesity. We examined the effects of leptin on platelet aggregation in morbidly obese subjects (n = 40; BMI, 41.6 +/- 1.1 kg/m2; leptin, 49.7 +/- 3.4 ng/ml) in comparison to normal-weight controls (n = 36; BMI, 23.3 +/- 0.4 kg/m2; leptin, 6.5 +/- 0.7 ng/ml). The aggregatory response to increasing concentrations of adenosine diphosphate (ADP) (2, 3, 4, and 5 microM) was significantly increased in platelets from obese compared to lean donors, reflecting a left shift in the dose-response curve. Plasma leptin levels, but not BMI, were significantly higher in subjects with stronger (above the median) compared to weaker (below the median) platelet aggregation at all ADP concentrations tested. In further experiments, stimulation (preincubation) with leptin (500 ng/ml) promoted ADP-induced platelet aggregation by approximately 25%, and there was no difference between platelets from obese and those from lean donors regarding the responsiveness to leptin (p = 0.99). Western blotting revealed that leptin induced phosphorylation of JAK2 and STAT3 to a similar extent in platelets from both groups. Expression of potential mediators of leptin resistance (SOCS3 and PTP1B) also did not differ in platelets from obese and control subjects. In conclusion, our data indicate that platelets from obese donors show increased aggregatory response to ADP, and that this might partly be the consequence of increased circulating leptin levels. Platelets from obese donors are not resistant to the enhancing effects of leptin on ADP-induced platelet aggregation. PMID- 19132238 TI - Altered bioavailability of platelet-derived factor VIII during thrombocytosis reverses phenotypic efficacy in haemophilic mice. AB - Ectopic delivery of factor VIII (FVIII) to megakaryocytes (Mk) represents a viable approach for localized tenase generation by concentrating the FVIIIa/FIXa enzyme-cofactor complex onto activated platelet membranes. We utilized a core rat platelet factor 4 (PF4) promoter for Mk/platelet-restricted expression of human B domain-deleted (hBDD) FVIII within the background of a haemophilia A mouse (rPF4/hBDD/FVIII-/-). Platelets from rPF4/hBDD/FVIII(-/-) mice contained approximately 122 mU FVIII:C/1 x 10(9) platelets/ml with no detectable plasmatic FVIII:C, and with no effect on alpha-granule-derived platelet factor V/Va function. Paired tenase assays (+/- thrombin) confirmed that platelet (pt) FVIII (unlike platelet FV) required thrombin cleavage for complete activation. rPF4/hBDD/FVIII(-/-) mice exposed to a thrombocytotic stimulus (thrombopoietin, TPO) demonstrated a statistically-significant 66% reduction in molar ptFVIII activity with a non-significant reduction in total ptFVIII biomass. Decreased molar ptFVIII concentration correlated with loss of phenotypic correction as evaluated using a haemostatic tail-snip assay. Comparative studies using a transgenic mouse expressing human amyloid-beta-precursor protein (hAbetaPP) from the rPF4 promoter confirmed diminished hAbetaPP expression without affecting endogenous alpha-granule PF4, establishing generalizability of these observations. While Mk/platelet-released ptFVIII (unlike pFV) is proteolytically inactive, we also conclude that thrombocytotic stimuli negatively affect ptFVIII bioavailability and phenotypic efficacy, results which correlate best with molar ptFVIII concentration, and not systemically available ptFVIII. PMID- 19132240 TI - Heparin-induced thrombocytopenia (HIT): clinical and economic outcomes. AB - Heparin-induced thrombocytopenia (HIT) is an immune-mediated adverse drug reaction that occurs following exposure to unfractionated heparin (UFH) or low molecular-weight heparin (LMWH). HIT with thrombosis (HITT) can cause devastating venous thromboembolism or arterial clots, prolonged hospitalization, and increased costs. To explore the economic and clinical implications of HIT and HITT, we initiated a single-center patient registry. In this report, we describe patient characteristics, comorbidities, management strategies, clinical outcomes, and costs. We enrolled 349 hospitalized patients with an enzyme immunoassay confirmed diagnosis of HIT over a 40-month period. Patients were assessed for the primary outcome of 30-day mortality, as well as baseline characteristics, development of thrombosis, and the economic impact of HIT. The primary outcome measure was 30-day mortality and occurred in 58 (16.6%) patients, 40 (15.3%) in the HIT group versus 18 (20.7%) in the HITT group (p = 0.25). The frequency of HIT was greater in patients exposed to UFH than in patients exposed to LMWH (0.8% vs. 0.2%, respectively, p < 0.001). Both HIT and HITT patients who were exposed to UFH had higher hospital costs than those exposed to LMWH ($113,100 vs. $56,352, respectively, p < 0.001). HIT remains an important clinical problem with a high mortality rate and significant cost, regardless of development of thrombosis. Prospective controlled trials need to be conducted to determine the optimal strategy to reduce the frequency of HIT. PMID- 19132241 TI - Thrombotic events in high risk patients are predicted by evaluating different pathways of platelet function. AB - A higher rate of clinical events in poor clopidogrel and/or aspirin responders was documented by using different methods to measure platelet function, but no conclusive data about the appropriate methodology to explore platelet reactivity are available. A total of 746 patients included in the cohort of the RECLOSE trial who had successful drug-eluting stent implantation were assessed for post treatment residual platelet reactivity (RPR) in platelet-rich plasma by 10 microM adenosine 5'-diphosphate (ADP), 1 mM arachidonic acid (AA) and 2 microg/ml collagen-induced platelet aggregation and in whole blood by the PFA-100 system. At six-month follow-up, RPR by two stimuli (ADP and AA or ADP and collagen) and by three stimuli (ADP, AA and collagen) is significantly associated with higher percentage of primary (definite or probable stent thrombosis) and secondary (cardiac mortality and stent thrombosis) end-points than RPR by ADP, AA, collagen and PFA-100 system. According to the primary and secondary end points, the specificity values for RPR identified by two (ADP and AA:94%; ADP and collagen:97%) and three stimuli were higher with respect to RPR by ADP (88%), or RPR by AA (83%) or RPR by collagen (90%). The positive likelihood ratio values of RPR by three stimuli (9.55) or of RPR by ADP and collagen (8.08) were higher than those of RPR by ADP (2.59), by AA (2.05), by collagen (4.73), or by PFA-100 (2.63). This prospective study documents that the evaluation of platelet reactivity addressed to identify patients at risk of thrombotic events on dual antiplatelet treatment has to be carried out by methods able to explore different pathways. PMID- 19132242 TI - Enzymatically hydrolyzed low-density lipoprotein modulates inflammatory responses in endothelial cells. AB - There is evidence that low-density lipoprotein (LDL) is modified by hydrolytic enzymes, and that the product (E-LDL) induces selective production of interleukin 8 (IL-8) in endothelial cells. Since nuclear factor-kappaB (NF-kappaB) is a major regulator of IL-8 transcription, we studied its activation in endothelial cells treated with E-LDL. Unexpectedly, the modified lipoprotein not only failed to activate NF-kappaB, but completely blocked its activation by tumour necrosis factor-alpha (TNF-alpha) in EA.hy926-cells, as assessed by electrophoretic mobility shift assays and immunofluorescence. Inhibition occurred upstream of NF kappaB translocation, as inhibitor of NF-kappaB- (IkappaB)-phosphorylation was suppressed by E-LDL. In contrast to NF-kappaB, transcription factor activator protein-1 (AP-1) proved to be activated. Removal of free fatty acids present in E LDL obliterated both activation of AP-1 and inhibition of NF-kappaB. Chromatin immunoprecipitation revealed that phosphorylated c-jun, but not NF-kappaBp65 bound to the natural IL-8 promoter. Production of endothelial IL-8 and simultaneous modulation of NF-kappaB in response to hydrolyzed LDL might serve to protect the vessel wall and promote silent removal of the insudated lipoprotein. PMID- 19132243 TI - High glucose conditions induce upregulation of fractalkine and monocyte chemotactic protein-1 in human smooth muscle cells. AB - The major complication of diabetes mellitus is accelerated atherosclerosis that entails an inflammatory process, in which fractalkine and monocyte chemotactic protein-1 (MCP-1) play a key role. We investigated the effect of diabetes associated high glucose (HG) on these chemokines and signalling mechanisms involved in human aortic smooth muscle cells (SMC). Exposure of SMC to HG resulted in an increase of fractalkine and MCP-1 expression and the activated mitogen-activated protein kinase (MAPK) signalling pathway, a process associated with elevated oxidative stress. Transfection with decoy oligodeoxynucleotides identified the involvement of transcription factors activator protein 1 (AP-1) and nuclear factor kappa B (NF-kappaB) in the observed up-regulation of chemokines. The MAPK inhibitors blocked the phosphorylation of IkBalpha and c jun, indicating the role of MAPK in NF-kappaB and AP-1 activation in SMC under HG conditions. The up-regulation of MCP-1 and fractalkine was associated with increased adhesive interactions between HG-exposed SMC and monocytes. Treatment of HG-exposed SMC with peroxisome proliferator-activated receptors alpha (PPARalpha) activators (fenofibrate and clofibrate) resulted in a reduction of mRNA and protein expression of MCP-1 and fractalkine. In conclusion, HG upregulates the expression of fractalkine and MCP-1 in SMC leading to increased monocyte-SMC adhesive interactions by a mechanism involving activation of MAPK, activator protein-1 (AP-1) and NF-kappaB. The increased expression of these two pro-inflammatory chemokines and the ensuing increased adhesion between SMC and monocytes may trigger the inflammatory process associated with further vascular complications of diabetes. PMID- 19132244 TI - Melanoma cell adhesion can be blocked by heparin in vitro: suggestion of VLA-4 as a novel target for antimetastatic approaches. AB - The clinical benefit of heparin in cancer patients to prolong survival can be attributed to non-anticoagulant mechanisms. Since adhesion molecules are crucially involved in tumour cell metastasis, their inhibition offers an attractive approach for interfering with the metastatic cascade. Heparin is known to attenuate metastasis in a selectin-dependent manner and possesses a variety of additional effects that are thought to influence tumour cell dissemination, proliferation, and angiogenesis. We investigated the adhesion behaviour of B16F10 melanoma cells in vitro regarding selectin- and VLA-4/VCAM-1-mediated binding to get an insight into underlying mechanisms of melanoma cell metastasis. We show that B16F10 cells display binding ability to P- and L-selectin as well as to isolated platelets. In contrast, B16F10 cells did not adhere to immobilized P selectin under flow. This contributes to recent findings that elucidate a major role of platelet P-selectin for microemboli formation and thus, facilitating metastasis. In contrast, B16F10 cells adhered to endothelial cells under flow, which could partly be inhibited by a function-blocking anti-VCAM-1 mAb. To emphasize VCAM-1 function, we analyzed cell adhesion at immobilized VCAM-1 and observed an integrin dependency. Inhibition experiments reveal that heparin influences VLA-4-mediated binding pathways. By a combination of different techniques we prove that the site of heparin action is rather VLA-4 than VCAM-1. To our knowledge, this is the first time that heparin is shown to interfere with the VLA-4/VCAM-1 interaction leading to the suggestion of a novel heparin target. Our results may contribute to the understanding of how heparin exerts its anti metastatic activity. PMID- 19132245 TI - Extended perioperative thromboprophylaxis in patients with cancer. A systematic review. AB - We systematically reviewed the literature to compare the relative efficacy and safety of extended versus limited duration heparin for perioperative thromboprophylaxis in patients with cancer. We followed the Cochrane Collaboration systematic review methodology and searched MEDLINE, EMBASE, ISI the Web of Science, and CENTRAL. The outcomes of interest included mortality, symptomatic deep venous thrombosis (DVT), pulmonary embolism, and bleeding. We evaluated the quality of evidence by outcome using the GRADE approach. Of 3,986 identified citations, we included three randomized clinical trials using low molecular-weight heparin (LMWH). The quality of evidence for mortality, DVT, and major bleeding was low. There was no significant difference between extended (4 weeks) and limited duration thromboprophylaxis in terms of death at three months (relative risk [RR] = 0.49; 95% confidence interval [CI] 0.12-1.94), or major bleeding at four weeks (RR = 2.94; 95% CI 0.12-71.85). An extended regimen was associated with a significantly lower risk of asymptomatic DVT (RR = 0.21; 95% CI 0.05-0.94). No data was available for symptomatic DVT. In conclusion, there is limited and low-quality evidence that extended duration LMWH for perioperative thromboprophylaxis reduces DVT in patients with cancer undergoing major abdominal or pelvic surgery. More and better quality evidence is needed to justify extended regimens. PMID- 19132246 TI - Influence of haematocrit on international normalised ratio (INR) differences between a whole blood point-of-care coagulation monitor and reference prothrombin time in plasma. AB - The CoaguChek S system is designed for use in patient self-testing. In this system a whole blood sample without sodium citrate is applied to a test strip containing thromboplastin and iron oxide particles. The detection principle is based on fibrin formation which inhibits and finally stops the movement of the iron oxide particles. In the classic prothrombin time (PT) test, citrate plasma is mixed with thromboplastin and an excess of calcium ions. In the monitoring of vitamin K-antagonist (VKA) treatment, all results are expressed on a common scale, i.e. international normalised ratio (INR). In patients on long-term VKA treatment, INRs were determined by the CoaguChek S system and reference methods for the classic PT. Four different CoaguChek S strip lots were evaluated. The difference in INR between the reference PT and the CoaguChek S system was negatively correlated to the haematocrit of the patients. We conclude that INR differences between CoaguChek S and plasma PT may be explained in part by the haematocrit. The magnitude of the effect of haematocrit, within the reference interval of 0.37 - 0.51, on the INR difference was not greater than approximately 10% for the combined data of the four strip lots. A bias of less than 10% seems to be acceptable clinically. PMID- 19132247 TI - Recombinant production of a hybrid plasminogen activator composed of surfactant protein B and low-molecular-weight urokinase. AB - Intraalveolar fibrin deposition is commonly observed during acute inflammatory and chronic interstitial lung diseases and may contribute to impairment of surfactant function and gas exchange. We recently described a chemically cross linked chimeric protein consisting of surfactant protein (SP)-B and urokinase (uPA) for targeting alveolar fibrin under conditions such as acute respiratory distress syndrome (ARDS) or lung fibrosis. We now investigated the feasibility of a recombinant production of a fusion protein encoding mature SP-B and uPA, termed SPUC. Four different SPUC proteins (N-term SP-B/C-term uPA, N-term uPA/C-term SP B, each +/- His-tag) were prepared by cloning the cDNA encoding mature SP-B and low-molecular-weight uPA into the expression vector pcDNA3.1. CHO-cells were transfected with the constructs and the supernatant and cell lysates were analyzed for expression of SPUC. Using a chromogenic substrate assay uPA activity was found in supernatants and lysates of transfected cells with highest activities related to the N-term uPA/C-term SP-B (+/- His-tag) construct in supernatants 48h after transfection. Casein enzymography showed an enzymatically active fusion proteins with a molecular weight of approximately 42 kDa in the supernatant of cells transfected with the N-term uPA/C-term SP-B (+/- His-tag) construct, but only a minor activity with the N-term SP-B/C-term uPA construct. The N-term uPA/C-term SP-B construct was also shown to possess higher resistance towards inhibition by plasminogen activator inhibitor-1. We conclude that recombinant production of a fusion protein consisting of mature SP-B and uPA is feasible, when the SP-B moiety is fused to the C-terminus of urokinase. PMID- 19132248 TI - Retrievable filter as an unusual cause of abdominal and lower back pain: a report of an exceptional case. PMID- 19132249 TI - Long-term efficacy of surgical removal of pseudomembranes in a child with ligneous conjunctivitis treated with plasminogen eyedrops. PMID- 19132250 TI - Three cases of abnormal fibrinogens: sumperk (Bbeta His67Leu), Unicov (Bbeta Gly414Ser), and Brno (gammaArg275His). PMID- 19132251 TI - Platelet surface P-selectin expression is highly correlated with serotonin transporter density in human subjects. PMID- 19132252 TI - Excluding pulmonary embolism without imaging tests--can our diagnostic algorithm be optimized? PMID- 19132253 TI - Fresh frozen plasma does not reduce in vivo thrombin formation after neonatal cardiopulmonary bypass surgery. PMID- 19132254 TI - Futile D-dimer testing in hospitalized patients--description, interpretation, improvements. PMID- 19132255 TI - TUBB1 Q43P polymorphism does not protect against acute coronary syndrome and premature myocardial infarction. PMID- 19132256 TI - On the significance of marginally low von Willebrand factor. PMID- 19132257 TI - Editor's choice articles 2006--2008. PMID- 19132258 TI - [Coding of hospital hyponutrition: the validity of a statement]. PMID- 19132259 TI - [Sociological contributions to the study of human nutrition: a developing scientific perspective in Spain]. AB - Human eating becomes one of the clearest intersections among biological, social, psychological and cultural processes nowadays. However, each scientific discipline has developed its own hypothesis and corpus of knowledge about human nutrition. Against this, it is the contention of this article to offer to nutritional professionals the efforts of sociology on this topic. It will be developed its purposes and contributions, its fields of research and, finally, a description of the social features of nourishment in modern societies. PMID- 19132260 TI - [SENPE-SEDOM document on coding of hospital hyponutrition]. PMID- 19132261 TI - [Bibliometric study of the journal Nutricion Hospitalaria for the period 2001- 2005: Part 2, consumption analysis; the bibliographic references]. AB - OBJECTIVE: To describe and assess the consumption of the information consulted and cited in the articles published in the journal Nutricion Hospitalaria for the period 2001--2005 by means of bibliometric analysis. METHOD: Cross-sectional descriptive analysis of the results obtained from the analysis of the lists of bibliographic references of the articles published at Nutricion Hospitalaria. We studied the most cited journals, the signatures index, the type of document referred, the publication language, the distribution of geographical origin, and obsolescence and readiness index. We took into account all types of documents with the exception of Communications to Congresses. RESULTS: 345 articles were published at Nutr Hosp, containing 8,113 bibliographic references, with a median of 18, a maximum of 136 and minimum of 0 BR per article. The mean (rate of publications per published article during the specified period) is 23.52 (95% IC 20.93-26.10) and the mean at 5% is 20.66 per article. The 25th and 75th percentiles are 6 and 32, respectively, the interquartile interval being 26 BR per document. The semi-period of Burton and Kebler is 7 years and the Price Index is 38.18%. CONCLUSION: The bibliographic references, the consumption of information, of the articles published at Nutricion Hospitalaria present parameters similar to other journals on health science. However, good data on obsolescence are observed, which reveal the good validity of most of the references studied. PMID- 19132262 TI - Antioxidant capacity of infant fruit beverages: influence of storage and in vitro gastrointestinal digestion. AB - AIM: The total antioxidant capacity of three beverages based on fruit juice, milk and cereals, intended for infants and young children up to 3 years of age was evaluated by two methods Trolox Equivalent Antioxidant Capacity and Oxygen Radical Absorption Capacity. RESULTS: According to the total antioxidant values obtained by both methods, the beverages can be ranked as follows: grape-orange banana > peach-apple > pineapple-banana. Ascorbic acid was the main contributor (60%) to the total antioxidant capacity, while the contribution of skimmed milk was less than 1.2%. After one month of storage at -20 degrees C, significant losses (p < 0.05) in total antioxidant capacity were found, though these were lower than 3% and therefore lacked nutritional significance. The bioaccessible fractions (maximum soluble fraction in simulated gastrointestinal media) of the beverages, obtained by in vitro gastrointestinal digestion, had antioxidant activities significantly lower (p < 0.05) than the original beverages, though the loss of antioxidant activity was always lower than 19%--thus indicating the stability of the total antioxidant capacity under the applied conditions. CONCLUSIONS: The total antioxidant capacity values of the bioaccessible fraction show that most antioxidants are available for absorption after digestion, and might contribute to the beneficial effects attributed to antioxidants. PMID- 19132263 TI - [Energy expenditure of healthy Brazilian adults: a comparison of methods]. AB - There are many available methods to measure the human energy consumption. Due to the high cost and small number of laboratories able to perform it, the direct calorimetry and doubly labeled water technique are rarely used to measure the energy expenditure. As consequence, there is an increase use of the indirect calorimetry and bioelectrical impedance analysis methods by researchers and predictive equations by physicians to calculate the total energy requirements in research and in clinic. OBJECTIVE: To evaluate the basal energy expenditure (GEB) and total energy expenditure (GET) of healthy subjects comparing results of indirect calorimetry (CI), bioelectrical impedance analysis (BIA) and predictive equations such as estimated energy requirements (EER), Fleish, FAO/WHO/UNU 1985 and 2001. MATERIAL AND METHODS: It was evaluated the energy expenditure in 81 subjects of both gender by CI and BIA. Of these, the GEB by predictive equations FAO/WHO/UNU 1985 and 2001 and the GET by EER, Fleisch and FAO/WHO/UNU 1985 and 2001 were calculated. The data was analyzed with the software Sigma Stat for Windows version 2.03 and Dunn's test was used for comparison of mean obtained with methods and equations (p < 0.05). RESULTS: It was verified that 60.5% of the participants were men. The majority of subjects were eutrofic. BIA showed significant differences between men and women. The differences between the values of GEB, obtained by BIA and CI, were statistically significant only for women (p < 0.05) with an underestimation of the GEB results obtained by BIA in relation to those obtained by CI. Significant difference was found between the predictive equations and CI for both sexes, also demonstrating an underestimation of the equations in relation to CI. It was observed an overestimation of GET with the predictive equations of FAO/WHO/UNU/85 and 2001 in relation to EER and Fleisch for both sexes. CONCLUSION: The estimation of GEB can be carried out by both BIA and CI, although BIA must be used with criteria since it underestimates in a significant way for women. In relation to the predictive equations, overestimation can develop positive energy balance in populations with risks for overweight and obesity. Thus, it is necessary comparisons to identify the limitations of each method. PMID- 19132264 TI - [Nitrogen losses and nitrogen balance in patients with malignant infarction of the median cerebral artery submitted to moderate hypothermia (32-33 degrees C)]. AB - OBJECTIVES: Induction of moderate hypothermia in patients with median cerebral artery (MCA) infarction may produce metabolic and nutritional impairments. Currently, we do not know which is the best method to carry out nutritional assessment in this population group. The aim of the present study was to assess the usefulness of nitrogen balance in the follow-up of patients with MCA submitted to moderate hypothermia (32-33 degrees C) by means of intravascular cooling at the Neurocritical Patients Unit at a tertiary hospital. MATERIAL AND METHODS: We designed a retrospective study including patients with MCA infarction of whom we gathered bio-demographical, clinical, hypothermia, and nutritional variables. Similarly, we carried out a prospective follow-up of a patient with MCA infarction and induced hypothermia, gathering the same variables at different time points of his clinical course. RESULTS: Six patients with MCA infarction submitted to moderate hypothermia for a mean duration of 12 days (interval 9-15) were included in the retrospective series. We observed that nitrogen losses (mean 9.9 g) were lower than those previously thought for critical patients during the acute phase. During the prospective follow-up of the patient with malignant infarction of the MCA from day 1 to day 22 after the application of hypothermia, low levels of nitrogen losses were similarly observed during the phase of induced hypothermia, which increased later on when the patient recovered normothermia. The mean nitrogen expenditure during the period of hypothermia was 10.7 g and increased up to 27.3 g during the normothermia period (day 17). CONCLUSIONS: These results suggest that moderate hypothermia-induced metabolic suppression is clinically relevant and thus the determination of nitrogen balance does not seem to be a useful tool in the nutritional followup of this type of patients. PMID- 19132265 TI - [The contribution of diet and sun exposure to the nutritional status of vitamin D in elderly Spanish women: the five countries study (OPTIFORD Project)]. AB - Vitamin D deficiency represents an important public health problem, especially among elderly people, by increasing the morbimortality. Because of the importance of this, in the year 2001 the "Five Countries Study" was put in place, within the European OPTIFORD project (Towards a strategy for optimal vitamin D fortification). This cross-sectional and observational study aims at knowing the vitamin D status in adolescent and elder women from five European countries, including Spain, according to different dietary and behavioral habits. OBJECTIVE: This work analyzes the relative contribution of sun exposure and diet to the vitamin D status in 53 Caucasian Spanish elder women (72 +/- 1.6 years), with an autonomous life, participating at the Five Countries Study. METHODOLOGY: The information was gathered in summer and winter time, by using homologated and validated questionnaires: health status and life style questionnaire; assessment of sun exposure by a standardized test and using a Viospor UV dosimeter (only in summer time); biochemical analysis of 25 hydroxyvitamin D (S-25-OHD) and parathyroid hormone; and questionnaire of frequency of selective intake. RESULTS: We observed a direct relationship between sun exposure measured with the dosimeter (741 +/- 624 J/m2) and the number of hours outdoors during the measuring week (3.4 +/- 1.9 hours/day) (p < 0.0001). The serum levels of S-25-OHD were higher in summer time than during the winter (40.32 +/- 20.39 nmol/L and 30.08 +/- 17.39 nmol/L, respectively), and 40% of the participants had worse vitamin status in winter as compared with summer time (p < 0.001). During the summer time, vitamin D deficiency (S-25-OHD < or = 25 nmol/L) affected 28% of the population, being virtually twice as much during the winter time. The higher the sun exposure, as assessed by the dosimeter, the higher the value of S-25-OHD, with clear differences between participants in the S-25-OHD < or = 25 nmol/L group and those in the S-25-OHD > 50 nmol/L (p = 0.01). There is also a direct association between the number of hours outdoors and S-25-OHD (p = 0.09), with differences between the participants in the S-25-OHD < or = 25 nmol/L group and the S-25-OHD > 50 nmol/L group. In more than 95% of the sample, mean dietary intakes of vitamin D (5.17 +/- 4.84 microg/day in summer time and 4.70+/- 4.72 microg/day in winter time), the main source of which being fish, did not cover the Recommended Allowances. We did not observe a relationship between the dietary intake and blood levels of vitamin D. By contrast, those participants taking vitamin D supplements presented higher S-25-OHD levels (summer = 69.64 nmol/L and winter = 55 nmol/L) than those not consuming it (summer = 36.83 nmol/L and winter = 25.82 nmol/L) (psummer =0.0003 and p winter < 0.001). CONCLUSIONS: The deficient status of vitamin D among the elderly female population has to be corrected, whenever possible, with appropriate sun exposure and an increase in vitamin D intake through the diet, assessing at each particular case the benefits of pharmacological supplementation. PMID- 19132266 TI - Nutritional status of patients with gynecologic and breast cancer. AB - OBJECTIVE: To identify the preoperative nutritional status of women with gynecologic or breast cancer, in correlation with disease site and staging as well as previous treatments. SUBJECTS AND METHODS: A cross-sectional study of 250 women evaluated by Body Mass Index (BMI) and Subjective Global Assessment (SGA). For data analysis, the chi-square test was applied. RESULTS: Breast cancer was the most frequent cancer, predominating in 56.2%. The median age of the patients was 52 years. In about 57% of these women, the tumor was restricted to clinical stages 0, I and II and 77% of the women had not undergone any other oncologic treatment prior to surgery. Subjective Global Assessment detected 76% of nourished women and 24% undernourished women, while Body Mass Index identified 34% of nourished women, 3.6% undernourished women and 62.4% overweight/obese women. A low level of diagnostic agreement between normal nutrition and malnutrition by both methods was observed (63.8%; kappa (95% CI) = 0.0884 (-0.07 0.24). No correlation between nutritional evaluation and previous treatment and disease staging was observed. Concerning anatomic site, it was subjectively observed that women with cancer of the uterine corpus were more malnourished than the rest (p = 0.02). CONCLUSIONS: The findings suggest that a more careful evaluation should be employed to identify preoperative nutritional status in women with gynecologic or breast cancer. PMID- 19132268 TI - Effects of dietary supplementation with medicinal fungus in fasting glycemia levels of patients with colorectal cancer: a randomized, double-blind, placebo controlled clinical study. AB - OBJECTIVE: The objective of this study was to evaluate the effects of dietary supplementation with Agaricus sylvaticus medicinal fungus in fasting glycemia of post-surgery patients with colorectal cancer. SCOPE: Proctology Ambulatory of Federal District Base Hospital-Brazil. SUBJECTS: Fifty-six colorectal cancer patients in postsurgery phase, stadiums phase I, II and II, which followed specific criteria of inclusion and exclusion. INTERVENTIONS: All patients were randomized in two groups: supplemented with Agaricus sylvaticus fungus (30 mg/kg/day) and placebo, and were followed up for six months. Subjects were divided later on according to BMI--Body Mass Index, sex and stage. Three fasting glycemia evaluations were carried out throughout the treatment. RESULTS: Subjects presented BMI medium = 24.65 kg/m2 (57.1% women and 42.9% men). The placebo group (average age 59.14 +/- 12.95 years) had initial glycemia levels of 94.36 +/- 15.34 mg/dL, after three months 98.12 +/- 15.54 mg/dL (p = 0.03) and in the sixth month 98.52 +/- 9.03 mg/dL (p = 0.01). The supplemented group (average age of 56.34 +/- 15.53 years) had initial glycemia levels of 95.92 +/- 11.64 mg/dL, after three months 94.88 +/- 12.24 mg/dL (p = 0.65) and, in the sixth month, a significant reduction to 92.86 +/- 6.82 mg/dL (p = 0.01). CONCLUSION: The results suggest that the dietary supplementation with Agaricus sylvaticus medicinal fungus can significantly reduce fasting glycemia levels of colorectal cancer patients in post-surgery phase. PMID- 19132267 TI - [Study of maternal nutrition and genetic on the foetal adiposity programming (The PREOBE study)]. AB - BACKGROUND: Maternal genetics and feeding before and during pregnancy, different maternal metabolic pathologies, as well as nutrient intakes of newborns in their first months of life may be involved in the obesity aetiology and its long-term consequences. The possible role of these and others factors, the mechanisms and the effects on the metabolism, and the development of this disease need further research. OBJECTIVE: To acquire more knowledge about foetal adipose tissue development and the influence of genetic, dietetic and environmental factors on the risk to suffer from obesity. METHODOLOGY: Four study groups have been established with 30 pregnant women in each one: 1) control group; 2) mothers with glucose intolerance/gestational diabetes; 3) women with low weight gain during pregnancy, and 4) women with overweight/obesity at the beginning of the pregnancy. The magnitudes to be studied are: 1) dietary intake; 2) life-style habits; 3) physical activity; 4) anthropometry and body composition; 5) haematological study; 6) biochemical study (lipid and metabolic biomarkers); 7) immune function profile related to nutritional status; 8) psychological profile; 9) genetic biomarkers, and 10) microbiological markers; all of them in relation to the development of the foetal adipose tissue in the first stages of life and the risk of suffering from obesity in the future. CONCLUSION: This project, coordinated by the Department of Paediatrics of the School of Medicine in the University of Granada, and with the collaboration of well-known and expert research groups, tries to contribute to the knowledge about the obesity aetiology in infancy and its subsequent development in later periods of life. PMID- 19132269 TI - Evolution of nutritional status of pediatric in patients of a tertiary care general hospital in Brazil. AB - JUSTIFICATION AND OBJECTIVE: Identify changes in the nutritional status of hospitalized children is fundamental for the early establishment of interventions. This study aims at describing the prevalence of undernutrition at admission and over the weeks of in-hospital stay in pediatric patients and evaluate the association between nutritional status and length of in-hospital stay. MATERIALS AND METHODS: A cohort study was carried out. It followed all the in-patients admitted to the general pediatric unit, composed of 72 beds, in the Hospital de Clinicas de Porto Alegre (HCPA), in the south of Brazil, from 20 march to 20 october in 2004. Patients who were between 1 month and 12 years of age and who had been admitted for clinical or/and surgical reasons were included. Those with Down Syndrome or without clinical condition and/or stature for weight measurement were excluded. Anthropometric data were collected up to 48 hours after admission and, weekly, up to hospital discharge (at admission, on 7th, 14th, and 21st day after admission). In children below 5 years of age, the standard defined by the World Heath Organization (WHO/2006) for the classification of the z-score for the stature/age (S/A), weight/age (W/A) and weight/stature (W/S) scores was used. In children from 5 to 10 years of age, the standards of the National Center for Health Statistics (NCHS, 1977) were used to classify the same rates as reference values. In children above 10 years of age, the classification of the Body Mass Index (BMI) was used (OMS/1995). In order to compare the z-scores over the four evaluation moments, analysis of variance (ANOVA) was used for repeated measurements, with Bonferroni's Post-Hoc test, and, for the evaluation of the in-hospital stay length, according to the nutritional status, Kaplan-Meier's survival curve, in the SPSS program, version 12.0, was used. RESULTS: 426 patients were included in the study. 57% of them were male and 50.7% were below one year of age. At admission, the prevalence of malnutrition was 10%, 18%, 21% and 14.7%, according to the W/S, W/A, S/A, and BMI criteria, respectively. Improvement of the nutritional status over the in-hospital stay was observed (at admission to 21st day) in children below 5 years (Z-score W/A: from 1.49 +/- 2.47 to -0.85 +/- 2.36, p = 0.001, and S/A: from -1.69 +/- 2.05 to -1.21 +/- 1.99, p = 0.007) and also in 5 to 10 years of age (S/A: from -0.43 +/- 1.31 to -0.30 +/- 1.37, p = 0.024). Undernourished patients, compared to nourished patients (according to the W/S rate for children below 10 years of age and BMI for above 10 years of age), showed a higher probability of remaining hospitalized (HR = 1.41; IC95%: 1.02-1.92). CONCLUSIONS: The prevalence of malnutrition by the W/S score was about half the prevalence found by the W/A and W/A indices, possibly reflecting a chronic impairment of the nutritional status. Undernutrition was confirmed as a health problem, once undernourished patients remained hospitalized for longer periods, which is a treatable problem as it was observed an improvement in the nutritional status rates over in-hospital stay. PMID- 19132270 TI - Effects of soy milk as a dietary complement during the natural aging process. AB - INTRODUCTION: Oxidative stress is one mechanism that could contribute to the acceleration of aging and age-related diseases. On the other hand, because of their antioxidative qualities soybean derived foods could have beneficial effects on the aging process. OBJECTIVES: The aim of our work was to study the effects of a diet supplemented with soy milk on certain biological features of aging in rats. METHODS: Male Wistar rats of 3 to 18 months of age, were assigned to one of two diets: 1) Experimental Group, commercial rat formula and soy milk; 2) Control Group, commercial rat formula and water. Every three months both lipid profile and lipid peroxidation were determined and neuronal cells of hippocampus were counted in control and experimental rats. RESULTS: The soy milk diet significantly improved the plasmatic lipid profile, decreasing serum cholesterol (total as well as LDL) and serum triglycerides, HDL-cholesterol was significatively higher in experimental animals. The LDL/HDL ratio was thus significantly lowered. The soy diet also produced decreased values of lipid peroxidation in brain, liver and kidney. These effects were significant after 6 to 9 months. The experimental animals lost fewer hippocampal neurons than the controls. Finally at 18 months of age, a greater number of surviving animals in experimental group with respect to the control one was observed. CONCLUSIONS: 1) soy intake could have beneficial effects as a complement of normal diet, but not as a replacement for animal proteins and 2) these effects are the result of a very long period (almost lifelong) of consumption of this diet. PMID- 19132271 TI - Validation of predictive equations for weight and height using a metric tape. AB - Weight and height measurements are important data for the evaluation of nutritional status but some situations prevent the execution of these measurements in the standard manner, using special equipment or an estimate by predictive equations. Predictive equations of height and weight requiring only a metric tape as an instrument have been recently developed. OBJECTIVE: To validate three predictive equations for weight and two for height by Rabito and evaluating their agreement with the equations proposed by Chumlea. METHODS: The following data were collected: sex, age and anthropometric measurements, ie, weight (kg), height (m), subscapular skinfold (mm), calf (cm), arm (cm) and abdominal (cm) circumferences, arm length (cm), and half span (cm). Data were analyzed statistically using the Lin coefficient to test the agreement between the equations and the St. Laurent coefficient to compare the estimated weight and height values with real values. RESULTS: 100 adults (age 48 +/- 18 years) admitted to the University Hospital (HCFMRP/USP) were evaluated. Equations I: W(kg) = 0.5030 (AC) + 0.5634 (AbC) + 1.3180 (CC) +0.0339 (SSSF) - 43.1560 and II: W (kg) = 0.4808 (AC) + 0.5646 (AbC) +1.3160 (CC) - 42.2450 showed the highest coefficients of agreement for weight and equations IV and V showed the highest coefficients of agreement for height. The St. Laurent coefficient indicated that equations III and V were valid for weight and height, respectively. CONCLUSION: Among the validated equations, the number III W (kg) = 0.5759 (AC) + 0.5263 (AbC) +1.2452 (CC) - 4.8689 (S) - 32.9241 and VH (m) = 63,525 -3,237(S) - 0,06904 (A) + 1,293 (HS) are recommended for height or weight because of their easy use for hospitalized patients and the equations be validated in other situations. PMID- 19132272 TI - [Nutritional status and diet quality in a group of school dancers]. PMID- 19132273 TI - [Nutritional support of the critically ill patient with continuous renal replacement therapies]. PMID- 19132274 TI - Nutritional diagnosis obtained by subjective global assessment in surgical patients and occurrence of post operative complications. PMID- 19132275 TI - [Study of dietary habits of young sportsmen]. PMID- 19132276 TI - [Perception of obesity and culture]. PMID- 19132277 TI - [Factors associated to hyponutrition in elderly people attending health care and social-health care centers]. PMID- 19132278 TI - [Dietary model of college students]. PMID- 19132279 TI - [Comparison between subjective global assessment and body mass index in the evaluation of hospital hyponutrition]. PMID- 19132280 TI - Should the food guide be implemented without evaluation? PMID- 19132283 TI - Vertical fluxes and accumulation of organochlorine pesticides in sediments of Haihe River, Tianjin, China. AB - Organochlorine pesticides (OCPs) concentrations in sediments and sediment trap fluxes of particulate organic carbon and OCPs were measured from October 2006 to May 2008 in the urban reach of Haihe River, Tianjin, China, in order to investigate vertical fluxes and accumulation of OCPs in sediments. The concentrations of OCPs in sediments (dry weight basis) were in the range of 2.08 10.07 ng/g (mean 4.70 ng/g) for HCHs (alpha-, beta-, gamma-, delta-HCH), 5.82 43.39 ng/g (mean 17.43 ng/g) for DDTs (p,p'-DDE, p,p'-DDD, o,p'-DDT, p,p'-DDT). Compared with data reported in Haihe River 2003, the levels of OCPs showed a significant decrease between 2003 and 2008. This is mainly attributed to significant decrease of production and usage of HCHs and DDTs and biodegradation of HCHs and DDTs in sediment. Concentrations of HCHs and DDTs in sediments were well correlated with sediment organic carbon contents. Concentrations of OCPs in settling particles (dry weight basis) ranged from 21.2 to 138.4 ng/g (mean 52.6 ng/g) for HCHs, 30.0-245.7 ng/g (mean 87.1 ng/g) for DDTs, which were 2.2-48.2 fold higher than those associated with sediments. beta-HCH was the dominant HCH isomer in both sediments and settling particles. Among DDTs, p,p'-DDE was found to be dominant in sediments, while p,p'-DDT was the predominant form in settling particles. This suggests that further biodegradation of OCPs occurred during the course of sedimentation and burial. The annual sediment trap flux of OCPs in Haihe River in the urban area of Tianjin was 177-211 microg/m(2) yr for HCHs and 213-341 microg/m(2) yr for DDTs, respectively. PMID- 19132284 TI - Urogynaecology training in the UK: past, present and future. AB - INTRODUCTION: With the implementation of Modernising Medical Careers, there has been considerable discussion and debate regarding the role of the urogynaecologist in secondary and tertiary care and how trainees with subspecialty training and those with a special interest fit into the current and future provision of urogynaecology services within the framework of obstetrics and gynaecology as a whole. DISCUSSION: Because of these changes, training in urogynaecology has and will continue to see major changes. This will also have an impact on workforce planning and service provision within the NHS. An attempt to highlight these issues has been made in this article. PMID- 19132286 TI - Wheat streak mosaic virus genotypes introduced to Argentina are closely related to isolates from the American Pacific Northwest and Australia. AB - Wheat streak mosaic virus (WSMV) was first detected in Argentina in 2002. Comparison of 78 WSMV coat protein sequences revealed that three Argentine isolates were closely related to isolates from the American Pacific Northwest (APNW) and Australia. Complete sequences were determined for one Argentine isolate, four APNW isolates, and three additional isolates from other regions of the USA. Comparison of these eight new sequences with five previously sequenced isolates of WSMV confirmed close affinity of WSMV from Argentina with APNW isolates. Collectively, these results indicate concurrent establishment of the same WSMV lineage in both Argentina and Australia. PMID- 19132285 TI - Continued symptom improvement in sexually active women with overactive bladder and urgency urinary incontinence treated with tolterodine ER for 6 months. AB - Improvements in overactive bladder (OAB) symptoms and health-related quality of life (HRQL) were assessed during a 24-week study of tolterodine extended release (TOL ER) in sexually active women with OAB and urgency urinary incontinence (UUI). A 12-week, double-blind, randomized, placebo-controlled trial was followed by a 12-week open-label phase. Sexually active women reported symptoms for >or=3 months. Subjects completed bladder diaries and HRQL measures at baseline and weeks 12 and 24. One hundred sixty-one women received TOL ER for 24 weeks. Women reported significant improvements in all end points at week 12 that were maintained or improved at 24 weeks. At week 24, 70% of subjects reported no UUI episodes. TOL ER resulted in improvements in OAB symptoms and HRQL that were maintained or greater with 6 months of use. Long-term compliance with OAB pharmacotherapy may be important for optimal treatment outcomes. PMID- 19132287 TI - Proteomic characterization of the sulfur-reducing hyperthermophilic archaeon Thermococcus onnurineus NA1 by 2-DE/MS-MS. AB - Thermococcus onnurineus NA1, a sulfur-reducing hyperthermophilic archaeon, was isolated from a deep-sea hydrothermal vent area in Papua New Guinea. The strain requires elemental sulfur as a terminal electron acceptor for heterotrophic growth on peptides, amino acids and sugars. Recently, genome sequencing of Thermococcus onnurineus NA1 was completed. In this study, 2-DE/MS-MS analysis of the cytosolic proteome was performed to elucidate the metabolic characterization of Thermococcus onnurineus NA1 at the protein level. Among the 1,136 visualized protein spots, 110 proteins were identified. Enzymes related to metabolic pathways of amino acids utilization, glycolysis, pyruvate conversion, ATP synthesis, and protein synthesis were identified as abundant proteins, highlighting the fact that these are major metabolic pathways in Thermococcus onnurineus NA1. Interestingly, multiple spots of phosphoenolpyruvate synthetase and elongation factor Tu were found on 2D gels generated by truncation at the N terminus, implicating the cellular regulatory mechanism of this key enzyme by protease degradation. In addition to the proteins involved in metabolic systems, we also identified various proteases and stress-related proteins. The proteomic characterization of abundantly induced proteins using 2-DE/MS-MS enables a better understanding of Thermococcus onnurineus NA1 metabolism. PMID- 19132288 TI - Achyranthoside H methyl ester, a novel oleanolic acid saponin derivative from Achyranthes fauriei roots, induces apoptosis in human breast cancer MCF-7 and MDA MB-453 cells via a caspase activation pathway. AB - Achyranthoside H methyl ester (AH-Me) is an oleanolic acid saponin derivative isolated from the roots of Achyranthes fauriei through diazomethane treatment. AH Me exhibited significant cytotoxicity against human breast cancer MCF-7 and MDA MB-453 cells, with respective ID(50) values of 4.0 and 6.5 muM: in the MTT assay. AH-Me is a unique saponin containing three methoxycarbonyl groups in the sugar moiety linked to the C-3 position of oleanolic acid. The demethylation of these methoxycarbonyl groups by alkaline hydrolysis caused a marked reduction of the cytotoxicity of AH-Me, suggesting that the methoxycarbonyl groups of AH-Me are key groups for the acquisition of cytotoxicity against human cancer cells. The staining of cancer cells with 4',6'-diamidino-2-phenylindole (DAPI) showed that the population of cells with altered nuclear morphology, for example chromatin condensation and fragmentation, increased markedly after AH-Me treatment. Exposure of MCF-7 and MDA-MB-453 cells to AH-Me resulted in a dose-dependent and time-dependent increase in the sub-G1 population, and in the cleavage of poly-ADP ribose polymerase (PARP) followed by the formation of an 89 kD peptide. Pretreatment of the cells with the pan-caspase inhibitor z-VAD-fmk abolished the cleavage of PARP by AH-Me treatment and suppressed the antiproliferative effect of AH-Me on tumor cell growth. These results together led to the suggestion that AH-Me induces apoptosis via the caspase activation pathway in human breast cancer cells, and apoptosis is the major mode of the cytotoxic effect triggered by AH Me. PMID- 19132289 TI - A new ecdysteroside from Lygodium japonicum (Thunb.) Sw. AB - A new ecdysteroside, named lygodiumsteroside A, as well as a known ecdysteroside, were isolated from the roots of Lygodium japonicum (Thunb.) Sw. Their structures were identified by spectral and chemical methods. PMID- 19132290 TI - Breaking biological symmetry in membrane proteins: the asymmetrical orientation of PsaC on the pseudo-C2 symmetric Photosystem I core. AB - The elucidation of assembly pathways of multi-subunit membrane proteins is of growing interest in structural biology. In this study, we provide an analysis of the assembly of the asymmetrically oriented PsaC subunit on the pseudo C(2) symmetric Photosystem I core. Based on a comparison of the differences in the NMR solution structure of unbound PsaC with that of the X-ray crystal structure of bound PsaC, and on a detailed analysis of the PsaC binding site surrounding the F(X) iron-sulfur cluster, two models can be envisioned for what are likely the last steps in the assembly of Photosystem I. Here, we dissect both models and attempt to address heretofore unrecognized issues by proposing a mechanism that includes a thermodynamic perspective. Experimental strategies to verify the models are proposed. In closing, the evolutionary aspects of the assembly process will be considered, with special reference to the structural arrangement of the PsaC binding surface. PMID- 19132291 TI - Molecular domestication of transposable elements: from detrimental parasites to useful host genes. AB - Transposable elements (TEs) are commonly viewed as molecular parasites producing mainly neutral or deleterious effects in host genomes through their ability to move. However, during the past two decades, major interest has been focusing on the positive contribution of these elements in the evolution of gene regulation and in the creation of diverse structural host genes. Indeed, DNA transposons carry an attractive and elaborate enzymatic machinery as well as DNA components that have been co-opted in several cases by the host genome via an evolutionary process referred to as molecular domestication. A large number of transposon derived genes known to date have been recruited by the host to function as transcriptional regulators; however, the biological role of the majority of them remains undetermined. Our knowledge on the structure, distribution, evolution and mechanism of transposons will continue to provide important contributions to our understanding of host genome functions. PMID- 19132292 TI - Sugar-mimicking glycosidase inhibitors: bioactivity and application. AB - A large number of compounds mimicking the structures of monosaccharides or oligosaccharides have been discovered from natural sources. Such sugar mimics inhibit carbohydrate-degrading enzymes because of a structural resemblance to the sugar moiety of the natural substrate. Carbohydrate-degrading enzymes are involved in a wide range of important biological processes, such as intestinal digestion, posttranslational processing of the sugar chain of glycoproteins, their quality control mechanisms, lysosomal catabolism of glycoconjugates, and some viral infections. It has now been realized that inhibitors of the enzymes have enormous therapeutic potential in diabetes and lysosomal storage disorders. In this review, the general bioactivity, current applications, and the prospects for new therapeutic applications are described. PMID- 19132293 TI - Signaling versus punching hole: How do Bacillus thuringiensis toxins kill insect midgut cells? AB - Cry proteins, produced by Bacillus thuringiensis (Bt), are widely used for the control of insect pests in agriculture as spray products or expressed in transgenic crops, such as maize and cotton. Little was known regarding the mechanism of action of these toxins when the first commercial Bt product was introduced fifty years ago. However, research on the mechanism of action over the last two decades has enhanced our knowledge of toxin interaction with membrane receptors and their effects in insect midgut cells. All this information allowed for the rational design of improved toxins with higher toxicity or toxins that overcome insect resistance, which could compromise Bt use and effectiveness in the field. In this review we discuss and evaluate the different models of the mode of action of Cry toxins, including a discussion about the role of various receptors in toxin action. PMID- 19132294 TI - "Classical 3 + 3 design" versus "accelerated titration designs": analysis of 270 phase 1 trials investigating anti-cancer agents. AB - The number of patients treated at each dose-level in dose seeking phase I trials is arbitrarily established. The most frequently used design is the "classical 3 + 3 design (3 + 3D)". Recently, Simon et al. had introduced several "accelerated titration designs (ATD)". In the present analysis, we compared the performance of these two types of designs in 270 recently (1997-2008) published phase I trials. ATD had been used in only 10% of the recent studies. ATD had permitted to explore significantly more dose levels (seven versus five, p = 0.0001) and reduced the rate of patients treated at doses below phase-2 recommended dose (46% versus 56%, p = 0.0001). Nevertheless, ATD did not allow a reduction in the number of enrolled patients, shorten the accrual time nor increase the efficacy of phase I trials. These data support that ATD as an effective clinical trial design over a standard 3 + 3 dose escalation design. PMID- 19132295 TI - Anti-proliferative effects of simocyclinone D8 (SD8), a novel catalytic inhibitor of topoisomerase II. AB - INTRODUCTION: Simocyclinone D-8 (SD8), a semi-synthetic compound derived from yeast, has been shown to decrease the proliferation of MCF-7 breast cancer cells. It has been shown to be a potent bacterial DNA gyrase inhibitor, a homologue of human topoisomerase II (hTopoII). We tested SD8 activity alone and in combination with cisplatin against malignant mesothelioma (MM) and non-small cell cancer (NSCLC) cell lines. METHODS: Inhibition of hTopoII supercoiling function by SD8 and a known hTopoII poison, etoposide, were done by in vitro assay using purified hTopoII and kinetoplast DNA as the substrate. The DNA products were analyzed by agarose gel electrophoresis after treatment with increasing concentrations of each drug. Mesothelioma cell lines (H2373, H2461 and H2596) and NSCLC cell lines (H2030, H460, and H2009) grown in RPMI with 10% calf serum were used. Non malignant mesothelial cells, LP9, were grown in 1:1 ratio of MCDB:199E medium supplemented with 15% calf serum, 0.4 microg/mL hydrocortisone, and 15 ng/mL epidermal grown factor. Cell proliferation assays were performed in 96-well plates using the CCK-8 kit (Dojindo inc.). Cells were treated for 72 h with various SD8 concentrations and controls containing equal volume of the vehicle, DMSO. Treated cells were assayed for the induction of apoptosis with poly ADP ribose polymerase-1 (PARP) cleavage assay. RESULTS: Biochemical assays revealed that the IC(50) for hTopoII inhibition was 100 microM for SD8 and 400 microM for etoposide. SD8 inhibited hTopoII function without inducing DNA cleavage events. SD8 inhibited the growth of NSCLC and Mesothelioma cells with IC(50) ranging from 75-125 microM. Furthermore, SD8 was not toxic to non-transformed primary mesothelial cell line, LP9 at the IC(50) doses. SD8 induced apoptosis in all cell lines tested. CONCLUSIONS: SD8 inhibits hTopoII in vitro without inducing DNA strands breaks and has significant activity against NSCLC and MM cell lines. While doses required for SD8 anticancer activity are unlikely to be achieved in vivo, chemical modifications to SD8 to increase its potency could lead to improved therapies for these diseases. PMID- 19132296 TI - Excellent visual outcome following Aspergillus flavus endogenous endophthalmitis farmer's lung disease. AB - INTRODUCTION: The purpose is to report an eye with endogenous Aspergillus flavus endophthalmitis that achieved a good visual outcome following early and aggressive management. METHODS: A 76-year-old male recently hospitalized for allergic Aspergillus pneumonitis after cleaning out a grain bin presented with reduced vision and anterior chamber and vitreous inflammation. The patient was treated with intravenous amphotericin and a pars plana vitrectomy with intravitreal amphotericin, and the vitreous biopsy sent for histopathological and microbial analysis. RESULTS: A. flavus was isolated from the vitreous biopsy. Two weeks after vitrectomy, intravitreal amphotericin was again injected into the affected eye. The patient regained vision to 20/80 several months later, despite a moderate cataract. CONCLUSION: Early treatment of A. flavus endophthalmitis with pars plana vitrectomy, intravitreal and systemic amphotericin can lead to good visual outcomes. PMID- 19132299 TI - The contribution of the BELLA study in filling the gap of knowledge on mental health and well-being in children and adolescents in Germany. AB - The mental health of children and adolescents is an epidemiological outcome of considerable public health relevance. This is because of a comparatively high prevalence of mental health problems and disorders in children and adolescents. The 'BELLA Study on Mental Health', a modular component of the Health Interview and Examination Survey for Children and Adolescents conducted by the Robert Koch Institute, assessed the prevalence of general and specific mental health problems in a representative sample of children and adolescents in Germany. Furthermore, determinants of mental health were studied in a developmental context. In the cross-sectional part of the study, participants were interviewed to assess the occurrence of risk factors and available assets (protective factors) for mental health, providing valuable data for health monitoring and planning. The sample was also repeatedly studied in a 2-year longitudinal design in order to estimate the effect size of risk and protective factors for mental health, and to analyse their combined effects on the process of mental health development. The BELLA study--as described in this supplement--supplies valuable estimates of the prevalence of mental health problems and disorders, identifies risk groups needing intervention, and describes the subjective health of children and adolescents in Germany. PMID- 19132300 TI - The mental health module (BELLA study) within the German Health Interview and Examination Survey of Children and Adolescents (KiGGS): study design and methods. AB - The BELLA study on mental health and well-being in children and adolescents is the mental health module of the German Health Interview and Examination Survey for Children and Adolescents (KiGGS). The cross-sectional KiGGS survey collected comprehensive data on the health status of 17,641 children and adolescents, aged 0-17 years, living in Germany. The survey included physical examinations and tests, questionnaires filled in by parents and equivalent questionnaires for children aged 11 years or older. A computer-assisted personal interview was conducted by study physicians, and various laboratory tests, e.g. on blood and urine samples, were performed. The longitudinal BELLA study collected initial cross-sectional data from a representative KiGGS sub-sample of families with children aged 7-17 years. Of the 4,199 randomly selected families invited to participate in the BELLA study, 2,863 took part. A total of 48.5% of the participating children and adolescents were girls, 51.5% were boys. Within the BELLA study, trained interviewers conducted standardised telephone interviews with one parent and also the child, if it was at least 11 years old. Afterwards the families received additional questionnaires. The interviews and questionnaires included various standardised and ICD-10-oriented instruments examining overall mental health problems and associated burden, depression, anxiety, attention deficit-/hyperactivity and conduct disorders. Furthermore they covered a broad spectrum of aspects related to mental health and well-being, such as risk and protective factors and health-related quality of life. An analysis of the non-responders showed that the families who agreed to participate form a representative sample with respect to the German population. The sample deviated only slightly from the KiGGS sample structure regarding socioeconomic status and regarding parent-reported mental health problems. No such difference was observed for the children's self-reported mental health. A weighting procedure was applied to correct for deviations from the sociodemographic and socioeconomic structure of the target population. PMID- 19132301 TI - Prevalence of mental health problems among children and adolescents in Germany: results of the BELLA study within the National Health Interview and Examination Survey. AB - BACKGROUND: Over the past decades the public health relevance of mental health conditions in children and adolescents has been of growing concern. However, so far no detailed epidemiological data has been available for a representative national sample in Germany. OBJECTIVES: The present paper reports prevalence rates of general and specific mental health problems among children and adolescents in Germany and describes the link between symptoms and impairment as well as the treatment situation. METHODS: The mental health module (BELLA study) examines mental health problems in a representative sub-sample of 2,863 families with children aged 7-17 from the National Health Interview and Examination Survey among Children and Adolescents (KiGGS). Mental health problems were determined using the extended version of the strengths and difficulties questionnaire (SDQ). Further standardised screening measures were employed to screen for anxiety disorders (SCARED), conduct disorder (CBCL), attention deficit-/hyperactivity disorder (FBB-HKS, Conners' Scale) and depressive disorders (CES-DC). Furthermore, substance abuse and suicidal tendencies were assessed. Health related quality of life (HRQoL) and health care use were determined. RESULTS: Overall, 14.5% of the children and adolescents aged 7-17 fulfilled the criteria for at least one specific mental health problem associated with impairment, or had an overall mental health problem indicated by an abnormal SDQ score and present impairment. However, high comorbidity was found in the children concerned. Symptoms of overall mental health problems were present in 8.6% of the children and 6.6% of the adolescents. This number was reduced to prevalence rates of 6.3 and 4.9% when additional impairment was taken as a criterion. Irrespective of the type of disorder, fewer than half of the children affected were reported as receiving treatment. However, for those suffering from mental health problems, large impairments in HRQoL were observed. CONCLUSIONS: The observed prevalence of mental health problems as well as their large impact on well-being and functioning calls for early prevention. This is especially important with regard to the large decrease in HRQoL in the children and adolescents affected. PMID- 19132302 TI - Assessing psychopathological problems of children and adolescents from 3 to 17 years in a nationwide representative sample: results of the German health interview and examination survey for children and adolescents (KiGGS). AB - BACKGROUND: Reliable information on the prevalence of mental health problems in children and adolescents in the general population in Germany is scarce. With the German health and examination survey for children and adolescents (KiGGS) data is available for the first time on psychopathological problems for the complete age range from 3 to 17 years. OBJECTIVES: To estimate prevalence rates for several groups with broadly defined psychopathology and to report distributions of psychopathological problems for total and fine-grained subgroups according to age, gender, socioeconomic status (SES), migration of children and adolescents from 3 to 17 years. METHODS: The strengths and difficulties questionnaire (SDQ) parent version was completed for 14,478 children and adolescents from 3 to 17 years. Data was collected from May 2003 to May 2006 from 167 representative sample points all over Germany. Prevalence rates of SDQ scales were calculated. Effects of age, gender, socio-economic status and migration status were determined. RESULTS: A total of 18.5% of the boys and girls were classified as 'borderline or abnormal' in the total difficulties score and thus determined as risk group for public health policies. Analyses of Variance showed significant effects for age and gender, SES and migration status. Significant interactions between age and gender were found for the total difficulties, conduct problems, hyperactivity-inattention and the emotional symptoms scores; significant interaction between SES and migration status was found for the prosocial behaviour score. CONCLUSIONS: Prevalence rates of psychopathological problems in children and adolescents are in line with other published findings. Younger age, lower SES and migration are related to more psychopathological problems. While girls display more emotional problems, boys have more externalizing problems. PMID- 19132303 TI - How impaired are children and adolescents by mental health problems? Results of the BELLA study. AB - BACKGROUND: The consideration of impairment plays a crucial role in detecting significant mental health problems in children whose symptoms do not meet diagnostic criteria. The assessment of impairment may be particularly relevant when only short screening instruments are applied in epidemiological surveys. Furthermore, differences between childrens' and parents' perceptions of present impairment and impairing symptoms are of interest with respect to treatment seeking behaviour. OBJECTIVES: The objectives were to assess parent- and self reported impairment due to mental health problems in a representative sample of children and adolescents; to describe the characteristics of highly impaired children with normal symptom scores; and to investigate the associations between symptoms in different problem areas and impairment. METHODS: The mental health module of the German Health Interview and Examination Survey for Children and Adolescents (the BELLA study) examined mental health in a representative sub sample of 2,863 families with children aged 7-17. Self-reported and parent reported symptoms of mental health problems and associated impairment were identified by the extended version of the strengths and difficulties questionnaire (SDQ) in children 11 years and older. RESULTS: Considerable levels of distress and functional impairment were found with 14.1% of the boys and 9.9% of the girls being severely impaired according to the parental reports. However, self-reported data shows a reversed gender-difference as well as lower levels of severe impairment (6.1% in boys; 10.0% in girls). Six percent of the sampled children suffer from pronounced impairment due to mental health problems but were not detected by screening for overall symptoms. Childrens' and parents' reports differed in regard to the association between reported symptom scores and associated impairment with children reporting higher impairment due to emotional problems. CONCLUSIONS: The assessment of impairment caused by mental health problems provides important information beyond the knowledge of symptoms and helps to identify an otherwise undetected high risk group. In the assessment of impairment, gender-specific issues have to be taken into account. Regarding the systematic differences between childrens' and parents' reports in the assessment of impairment, the child's perspective should be given special attention. PMID- 19132304 TI - How often are German children and adolescents diagnosed with ADHD? Prevalence based on the judgment of health care professionals: results of the German health and examination survey (KiGGS). AB - BACKGROUND: Attention deficit-/hyperactivity disorder (ADHD) is a chronic disorder with a substantial lifelong impact on personal and social functioning, academic performance, and the health system in general. Extended knowledge regarding its epidemiology will help to optimise the distribution of health resources and support affected children and adolescents. OBJECTIVES: To report (1) the lifetime prevalence rates of ADHD in children and adolescents in Germany ages 3-17 years diagnosed by health care professionals, (2) the symptoms of hyperactivity and inattention in children and adolescents, and 3) the distributions and odds ratios for gender, age, socioeconomic status (SES), and history of migration. METHODS: Data were collected from May 2003 until May 2006 in 167 representatively selected sample points in Germany. A total of 17,461 children and adolescents (7,569 boys and 7,267 girls) were medically and physically examined, and their parents completed a self-administered questionnaire. Parent-reports of a lifetime ADHD diagnosis by a medical doctor or psychologist were taken as case definitions. Additional information was obtained via the parents from the strengths and difficulties questionnaire (SDQ) and also from trained observers. RESULTS: The overall lifetime prevalence of ADHD diagnosis was 4.8%. As expected, there was a significant gender difference between boys (7.7%) and girls (1.8%). Additionally, 4.9% of subjects had scores above threshold on the Inattention/ Hyperactivity subscale of the SDQ. As expected, a significant age effect was found for ADHD diagnosis (1.5% preschool age; 5.3% primary school; 7.1% secondary school). There were neither German east/west differences nor differences for rural versus urban areas. However, socioeconomic status was significantly associated with the prevalence of diagnosis (low SES: 6.4%, medium SES: 5.0%; high SES: 3.2%). CONCLUSION: The prevalence of diagnosed ADHD and the influence of its mediating factors found in our study are in line with those from other European countries, but our findings reflect a lower band of variation. PMID- 19132305 TI - How often do children meet ICD-10/DSM-IV criteria of attention deficit /hyperactivity disorder and hyperkinetic disorder? Parent-based prevalence rates in a national sample--results of the BELLA study. AB - BACKGROUND: There is a lack of representative prevalence rates for attention deficit-/hyperactivity disorder (ADHD) according to DSM-IV criteria and hyperkinetic disorder (HD) according to ICD-10 criteria for German subjects. OBJECTIVE: To report the results of analyses of categorical data on the prevalence rates of the symptoms of ADHD/HD and additional diagnostic criteria, as well as of the diagnoses of ADHD and HD according to symptoms and other diagnostic criteria, according to the ICD-10 and DSM-IV. Further, to report administrative prevalence rates of the diagnosis and rates of co-existing behavioural and emotional problems. METHOD: Within the BELLA module of the German Health Interview and Examination Survey for Children and Adolescents (KiGGS), a representative sample of parents of 2,452 children and adolescents aged 7-17 years completed an ADHD symptom checklist (FBB-HKS/ADHS) and additional questionnaires for the assessment of coexisting behavioural and emotional problems. RESULTS: The prevalence rates for the diagnoses of ADHD according to DSM-IV criteria were 5.0% and the rate for HD according to ICD-10 criteria was 1.0%. Higher prevalence rates were found in boys and in younger children. The addition of other diagnostic criteria (impairment, pervasiveness, onset, duration) resulted in a significant decrease of the prevalence rates of ADHD and HD to 2.2 and 0.6%, respectively. Higher prevalence rates were found in families of lower socioeconomic status and families from urban areas. The lifetime administrative prevalence rate was 6.5%. Children with ADHD had an increased risk for coexisting behavioural and emotional problems, especially for aggressive and antisocial behaviour problems, but also for anxiety and mood problems. CONCLUSION: The results of the national sample are in line with community studies in other countries. The effects of the additional diagnostic criteria of impairment, situational pervasiveness, symptom onset and symptom duration on the prevalence rates have to be considered in other epidemiological studies. PMID- 19132306 TI - Depressive symptoms of children and adolescents in a German representative sample: results of the BELLA study. AB - BACKGROUND: In Europe, a considerable proportion of children and adolescents is affected by depressive symptoms, impairing their everyday life and social functioning. OBJECTIVES: The aim of this paper is to provide an overview of the depressive symptoms in children and adolescents in Germany, addressing risk factors, comorbidity, and impact of depressive symptoms on everyday life. METHODS: In the BELLA study, the mental health module of the German Health Interview and Examination Survey for Children and Adolescents (KiGGS), a representative sample of young people aged 7-17 years was enrolled. Depressiveness, assessed by the CES-DC, as well as other mental health problems were examined in the context of risk and protective factors. RESULTS: Depressive symptoms showed high prevalence in parent- and self-reports. Higher depression scores were found in those with a high number of psychosocial risks existing in the family, and they decreased as the number of protective factors the children and adolescents had at their disposal increased. Although only half of the boys and girls with high depression scores were regarded as significantly impaired, all of them had a much higher risk for additional mental health problems. Furthermore, their health-related quality of life was limited compared to their peers who had low depression scores. CONCLUSIONS: To differentiate between clinically significant depression and milder forms, it is necessary to take into account the different perspectives of children and their parents. Prevention and intervention should acknowledge the widespread distribution of depressive symptoms in children and adolescents, the high comorbidity of depressive and other mental health problems and the impact of depression on the aspects of everyday life. PMID- 19132307 TI - Disordered eating behaviour and attitudes, associated psychopathology and health related quality of life: results of the BELLA study. AB - OBJECTIVE: To identify disordered eating behaviour and attitudes in a large representative population in order to determine the relationship with body weight status, and to assess associated psychopathology and health-related quality of life. METHODS: A total of 11-17 year-old adolescents (n=1,895) were randomly selected from the national representative sample of 17,641 families participating in the German Health Interview and Examination Survey for Children and Adolescents (KiGGS). Weight and height were assessed by trained staff. Mental health problems and health-related quality of life were examined by means of a telephone interview and different questionnaires. Eating disordered behaviour and attitudes were identified by the SCOFF, an instrument consisting of five questions originally developed to screen for eating disorders in clinical settings. RESULTS: About one third of the girls and 15% of the boys reported disordered eating behaviour and attitudes, which were most prevalent in overweight youth. There was a significant association between the presence of disordered eating behaviour and psychopathology, which was comprised of internalising and externalising behavioural problems. In addition, adolescents with disordered eating behaviour reported reduced quality of life. CONCLUSION: The high prevalence of disordered eating in the general population of Germany is of great concern. Health professionals should not only be aware of disordered eating in underweight adolescents, but in all youth, especially overweight individuals. Disordered eating behaviour is associated with a wide range of psychopathological and psychosocial concerns. Thus, youngsters engaging in disordered eating behaviour should also be explored for other serious mental or social problems. PMID- 19132308 TI - Self-mutilation and suicidal behaviour in children and adolescents: prevalence and psychosocial correlates: results of the BELLA study. AB - OBJECTIVE: To investigate the prevalence and psychosocial correlates of suicidal behaviour in a representative sample of children and adolescents in Germany. METHODS: Suicidal behaviour was assessed in the BELLA study in a sample of 2,863 families with children aged 7-17 using the corresponding questions from the child behavior check list and the youth self report. Self-reported as well as parent reported measures of overall mental health problems, anxiety, depression, aggressive and delinquent behaviour, attention deficit-/hyperactivity as well as health-related quality of life were also administered. RESULTS: Self-mutilation and/or suicidal attempts within the last six months were reported by 2.9% of the adolescents 11-17 years of age. Suicidal thoughts were reported by 3.8% of the same group of adolescents. The prevalence rates reported by the parents were 1.4% for self-mutilation and/or suicidal attempts and 2.2% for suicidal thoughts. The prevalence of parent-reported self-mutilation/suicidal attempts in children below 11 years of age was very low. Youth reporting suicidal behaviour were older than youth not reporting suicidal behaviour. Children and adolescents exhibiting suicidal behaviour reported significantly more general mental health problems, depressive symptoms, anxiety, and hyperactivity as well as lower health-related quality of life. CONCLUSIONS: There is a strong connection between suicidal behaviour and emotional and behavioural problems, especially with symptoms of depression, anxiety and hyperactivity. The association observed between attention deficit-/hyperactivity and suicidal behaviour requires further investigation. The differences in the extent of reported suicidal behaviour in adolescents between the self- and parent-ratings and the degree of confidentiality in the collection of the data are subjects for future research. PMID- 19132309 TI - Psychometric properties of the parent strengths and difficulties questionnaire in the general population of German children and adolescents: results of the BELLA study. AB - BACKGROUND: The strengths and difficulties questionnaire (SDQ) is a brief screening instrument that addresses negative and positive behavioural attributes of children and adolescents in the age range of 4-16 years and can be completed by parents, teachers, and as a self-report. Furthermore, the impact supplement of the extended SDQ surveys for perceived problems, impact, and burden. OBJECTIVE: This paper aims to examine the psychometric properties of the parent form and to investigate differences in the SDQ scores for sociodemographic and socioeconomic subgroups. Patterns of association with other measures of mental health and descriptive comparison with the first normative sample are also reported. METHODS: Within the BELLA study module of the German Health Interview and Examination Survey for Children and Adolescents (KiGGS), a total of 2,406 children and adolescents aged 7-16 years as well as their parents answered the items of the SDQ and the additional impact supplement. The internal consistency of scale responses was assessed via Cronbach's alpha (alpha). Likert scale assumptions of sufficient and similar item-total correlation and item variance were investigated. The factorial validity of the SDQ measurement model was tested by means of exploratory and confirmatory factor analysis. Mean score differences between males and females, age groups (7-10 years vs. 11-16 years), and socioeconomic status groups (Winkler index) were examined via ANOVA. RESULTS: Factor analysis provided an exact replication of the original five-factor SDQ subscale structure. All subscales were sufficiently homogeneous. The mean total difficulties and SDQ subscale scores of the BELLA sample did not differ from the first German normative data. Younger children were more impaired on various SDQ scales than older children, girls were more emotionally affected, and boys showed more externalising problems. CONCLUSIONS: The present study confirmed the validity and reliability of the parent-reported SDQ scale structure. The SDQ was found to be a valid and helpful questionnaire for use in the framework of an epidemiological survey. PMID- 19132310 TI - Psychometric properties of two ADHD questionnaires: comparing the Conners' scale and the FBB-HKS in the general population of German children and adolescents- results of the BELLA study. AB - BACKGROUND AND OBJECTIVE: To examine and compare the psychometric properties of two short screening instruments for children and adolescents suffering from attention deficit-/ hyperactivity disorder (ADHD). The Conners' Hyperactivity Index consists of ten items that assess symptoms of hyperactivity through self report and parents' proxy. The German ADHD Rating scale (FBB-HKS/ADHS) consists of 20 items that assess the severity and perceived burden of inattention, hyperactivity, and impulsiveness as defined by the ICD-10 and DSM-IV. METHODS: Within the BELLA module of the German Health Interview and Examination Survey for Children and Adolescents (KiGGS), the parents of 2,863 children and adolescents rated the Conners' Hyperactivity Index and the FBB-HKS. RESULTS: The internal consistency of item responses was assessed via Cronbach's alpha and showed that both instrument scores were able to obtain a reliable measurement. The factorial validity of the FBB-HKS measurement model as well as the unidimensionality of the Conners' scale was tested by means of exploratory and confirmatory factor analysis (EFA and CFA), indicating satisfactory goodness of fit for the FBB-HKS (RMSEA=0.06) and some deviation from the unidimensionality assumption of the Conners' scale. Stability of results across age could be confirmed with few exceptions. Mean scores differences were found between both sexes, age groups, and different socioeconomic status groups (Winkler-Index) with males, younger respondents, and children with low socioeconomic status displaying more ADHD related behaviour. Correlation coefficients between the two instruments' scores and other scales assessing emotional and behavioural problems hinted at convergent validity. CONCLUSION: Both instruments' scores showed reliability as well as factorial and convergent/discriminant validity. The pros and cons of the two instruments as well as for which purpose and under which circumstances one of the measures can be favoured must be considered prior to applying such a measure. PMID- 19132311 TI - The German version of the Centre for Epidemiological Studies Depression Scale for Children: psychometric evaluation in a population-based survey of 7 to 17 years old children and adolescents--results of the BELLA study. AB - OBJECTIVES: To examine the psychometric properties and test the theoretical quality of the German version of the Centre for Epidemiological Studies depression scale for children (CES-DC), a 20-item screening instrument measuring the frequency of parent- and self-reported depressive symptoms in children and adolescents. METHODS: Using a population-based, representative sample of n=2,863 7 to 17-year-old German children and adolescents, factorial validity were determined by means of linear structural equation modelling. Cross-sectional coefficients of reliability, inter-rater agreement as well as descriptive statistics of the scales were calculated. RESULTS: In a population-based German sample, the four-factor version of the CES-DC following Radloff (Appl Psychol Meas 1:385-401, 1977) is considered to have good factorial validity and stability across age and informant versions. The main problems of the questionnaire are the high item difficulties, strong floor effects of the scales and low cross sectional reliability, which are acceptable only for screening purposes. The low inter-rater agreement indicates that parental assessment can replace self assessment only to a limited degree. CONCLUSION: The strengths and weaknesses of the CES-DC are discussed taking previous data and comparable tests into consideration. Particular advantages are the existence of the parent-report form and the adult version, as well as its multifactorial structure. Parental assessment should be supplemented by self-report data whenever possible. PMID- 19132312 TI - Psychometric properties of the KINDL-R questionnaire: results of the BELLA study. AB - BACKGROUND: The concept of health-related quality of life (HRQoL) involves the respondents' perception of well-being and functioning in physical, emotional, mental, social, and everyday life areas. Research in the area of subjective health has resulted in the development of a multitude of HRQoL instruments that meet satisfying psychometric standards with regard to reliability, validity, and sensitivity of the scales. One frequently used generic measure for children and adolescents is the KINDL-R questionnaire developed by Ravens-Sieberer and Bullinger (Qual Life Res 7:399-407, 1998). METHODS: Within the representative sample of the BELLA study, analyses regarding psychometric properties (namely reliability as well as discriminant and construct validity) are performed. RESULTS: Psychometric testing of the KINDL-R questionnaire reveals good scale utilisation and scale fit as well as moderate internal consistency. Correlations with the KIDSCREEN-52 subscales are shown. Differences in KINDL-R scores exist between chronically ill and healthy children as well as between SDQ problem scores. CONCLUSION: The KINDL-R is a suitable instrument for measuring HRQoL in children and adolescents through self-report. The testing of the instrument in a representative sample of German children and adolescents as well as their parents provides reference values extending the potential of the KINDL-R questionnaire. PMID- 19132313 TI - Risk and protective factors for children's and adolescents' mental health: results of the BELLA study. AB - BACKGROUND: Since prevalence rates of mental health problems in children and adolescents are high and of considerable relevance to public health, determinants of mental health, such as risk and protective factors, are of special interest. OBJECTIVES: The present paper reports the frequencies and distributions of potential risk and protective factors and analyses their effects on children's mental health. METHODS: The BELLA study is the mental health module of the German National Health Interview and Examination Survey among Children and Adolescents (KiGGS). Mental health problems and their assumed determinants are examined in a representative sub-sample of 2,863 families with children and adolescents aged 7 17. In order to identify mental health problems, the extended version of the Strengths and Difficulties Questionnaire was administered. Data on psychosocial risk factors as well as on protective factors in terms of personal, familial and social resources were collected by questioning the parents and, from the age of 11 years upwards, the children themselves. RESULTS: Adverse family climate stands out particularly as a negative contributor to children's mental health. When several risk factors occur simultaneously, the prevalence of mental health problems increases markedly. Conversely, pronounced individual, family and social resources coincide with a reduced occurrence of mental health problems, especially in children with a limited number of risk factors. CONCLUSIONS: The results suggest that differential prevention strategies are needed depending on the risk level: in the low risk group, to which most children belong, effective prevention programmes should define strengthening resources as a key objective. In the smaller group of children with a high number of risk factors, more complex intervention designs are required, which must consider the reduction of risks as well as strengthening resources. PMID- 19132314 TI - Health-related quality of life in children and adolescents in Germany: results of the BELLA study. AB - BACKGROUND: The self-perceived health or health-related quality of life of children and adolescents is increasingly recognised as a relevant outcome in medical practice and public health research. Identifying children and adolescents with particularly low health-related quality of life allows for an early detection of hidden morbidity and health care needs. OBJECTIVES: The present study investigates health-related quality of life in children and adolescents in Germany. METHODS: In the Mental Health Module (BELLA study) of the German National Health Interview and Examination Survey for Children and Adolescents (KiGGS), the parents of 2,863 children and adolescents aged 7-17 years, and 1,700 children and adolescents aged 11-17 years completed the KINDL-R quality of life questionnaire. RESULTS: The reliability (Cronbach's alpha=0.86) and validity of the measurements using the parent-reported KINDL-R were confirmed. Means and percentiles were calculated for the total sample as well as for strata defined by age, sex, geographical region (east/west), migration status and socioeconomic status. Expected differences in health-related quality of life of children and adolescents from different social backgrounds and with different health statuses were demonstrated by differences in the KINDL-R scores (effect size d up to 1.29). CONCLUSION: This study provides representative, normative data (self report and parent-report) on the test scores of health-related quality of life (KINDL-R) for the population of children and adolescents in Germany in general, as well as in sociodemographic and socioeconomic subpopulations. PMID- 19132315 TI - Pediatric neurology. Editorial. PMID- 19132316 TI - Management of pediatric migraine: Current concepts and controversies. AB - Migraine in children and adolescents is a common condition. Emerging knowledge about the pathophysiology of migraine is leading to new targeted treatments toward specific mechanisms of migraine. This review explores the evidence for different treatments for pediatric migraine including the periodic syndromes of childhood that commonly represent precursors to migraine. PMID- 19132317 TI - Arterial ischemic stroke in children--recent advances. AB - Childhood stroke syndromes are an important cause of mortality and morbidity. This paper focuses on the recent advances in arterial ischaemic stroke beyond the neonatal period. Vascular risk factors are identified in the majority of children and guide both acute and longer term treatments, as well as determining prognosis. Contrary to popular belief many children have residual impairments encompassing a wide range of domains. National and international collaborations are facilitating an increase in the understanding of childhood stroke and have the eventual aim of conducting trials of potential therapeutic interventions. PMID- 19132319 TI - Management of septic shock: where do we stand? PMID- 19132318 TI - Traumatic brain injury in children: recent advances in management. AB - To define and discuss new developments in the field of pediatric traumatic brain injury (TBI). Review of several recent key studies on therapy since publication of the first U.S. traumatic brain injury guidelines in 2003. In addition, we discuss new developments in the use of biomarkers of brain injury in TBI diagnosis and also discuss recent advances in bedside neuromonitoring that may be helpful in the setting of pediatric brain injury. Important new information on optimal cerebral perfusion pressure management, cerebrospinal fluid drainage, decompressive craniectomy, hypothermia, biomarkers of brain injury along with advances in neuromonitoring are presented. The 2003 guidelines have stimulated important new research. This is reshaping bedside care. PMID- 19132320 TI - Genomics of olfactory receptors. AB - In many species, the sense of smell plays important roles in locating food, detecting predators, navigating, and communicating social information. The olfactory system has evolved complex repertoires of odor receptors (ORs) to fulfill these functions. Through computational data mining, OR repertoires of multiple species were identified, revealing a surprisingly large OR gene family in rodents and evolutionary fluctuation among different organisms. Characteristics of OR genes were explored through computational and experimental methods, showing a complicated gene structure and special genomic distribution. Utilizing high-throughput OR microarrays, expression profiles of the mouse and human OR repertoire were examined, their olfactory functions verified, and their zonal, ectopic and developmental expression determined. Variation in human smelling abilities results from different functional OR repertoires, variable expressional levels and polymorphisms in the copy number of the OR genes. These genomic approaches have both provided new data and generated new questions. PMID- 19132322 TI - Impact of TNF-R1 and CD95 internalization on apoptotic and antiapoptotic signaling. AB - Internalization of cell surface receptors has long been regarded as a pure means to terminate signaling via receptor degradation. A growing body of information points to the fact that many internalized receptors are still in their active state and that signaling continues along the endocytic pathway. Thus endocytosis orchestrates cell signaling by coupling and integrating different cascades on the surface of endocytic vesicles to control the quality, duration, intensity, and distribution of signaling events. The death receptors tumor necrosis factor receptor 1 (TNF-R1) and CD95 (Fas, APO-1) are known not only to signal for cell death via apoptosis but are also capable of inducing antiapoptotic signals via transcription factor NF-kappaB induction or activation of the proliferative mitogen-activated protein kinase (MAPK)/ERK (extracellular signal-regulated kinase) protein kinase cascades, resulting in cell protection and tissue regeneration. A clue to the understanding of these contradictory biological phenomena may arise from recent findings which reveal a regulatory role of receptor internalization and intracellular receptor trafficking in selectively transmitting signals, which lead either to apoptosis or to the survival of the cell. In this chapter, we discuss the dichotomy of pro- and antiapoptotic signaling of the death receptors TNF-R1 and CD95. First, we will address the role of lipid rafts and post-translational modifications of death receptors in regulating the formation of receptor complexes. Then, we will discuss the role of internalization in determining the fate of the receptors and subsequently the specificity of signaling events. We propose that fusion of internalized TNF receptosomes with trans-Golgi vesicles should be recognized as a novel mechanism to transduce death signals along the endocytic route. Finally, the lessons learnt from the strategy of adenovirus to escape apoptosis by targeting death receptor internalization demonstrate the biological significance of TNF receptor compartmentalization for immunosurveillance. PMID- 19132323 TI - FasL expression and reverse signalling. AB - FasL plays a central role in the induction of apoptosis within the immune system. It mediates activation-induced cell death (AICD) of T lymphocytes and contributes to the cytotoxic effector function of T and NK cells. Moreover, FasL is discussed as direct effector molecule for the establishment of immune privilege and tumour survival. Besides its death-promoting activity, FasL has been implicated in reverse signalling and might thus also play a role in T cell development and selection and the modulation of T cell activation. Considering these diverse functions, the overall FasL expression has to be tightly controlled to avoid unwanted damage. Based on an activation-associated transcriptional control, several post-transcriptional processes ensure a safe storage, a rapid mobilisation, a target-directed activity and a subsequent inactivation. Over the past years, the identification and characterisation of FasL-interacting proteins provided novel insight into the mechanisms of FasL transport, processing and reverse signalling, which might be exemplary also for the other members of the TNF family. PMID- 19132324 TI - Glucocorticoid-mediated apoptosis resistance of solid tumors. AB - More than a quarter of a century ago, the phenomenon of glucocorticoid-induced apoptosis in the majority of hematological cells was first recognized. More recently, glucocorticoid-induced antiapoptotic signaling associated with apoptosis resistance towards cytotoxic therapy has been identified in cells of epithelial origin, most of malignant solid tumors and some other tissues. Despite these huge amounts of data demonstrating differential pro- and anti-apoptotic effects of glucocorticoids, the underlying mechanisms of cell type-specific glucocorticoid signaling are just beginning to be described. This review summarizes our present understanding of cell type-specific pro- and anti apoptotic signaling induced by glucocorticoids. We shortly introduce mechanisms of glucocorticoid resistance of hematological cells. We highlight and discuss the emerging molecular evidence of a general induction of survival signaling in epithelial cells and carcinoma cells by glucocorticoids. We give a summary of our current knowledge of decreased proliferation rates in response to glucocorticoid pre- and combination treatment, which are suspicious to be involved not only in protection of normal tissues, but also in protection of solid tumors from cytotoxic effects of anticancer agents. PMID- 19132321 TI - Many checkpoints on the road to cell death: regulation of Fas-FasL interactions and Fas signaling in peripheral immune responses. AB - Interactions between the TNF-family receptor Fas (CD95) and Fas Ligand (FasL, CD178) can efficiently induce apoptosis and are critical for the maintenance of immunological self-tolerance. FasL is kept under strict control by transcriptional and posttranslational regulation. Surface FasL can be cleaved by metalloproteases, resulting in shed extracellular domains, and FasL can also traffic to secretory lysosomes. Each form of FasL has distinct biological functions. Fas is more ubiquitously expressed, but its apoptosis-inducing function is regulated by a number of mechanisms including submembrane localization, efficiency of receptor signaling complex assembly and activation, and bcl-2 family members in some circumstances. When apoptosis is not induced, Fas-FasL interactions can also trigger a number of activating and proinflammatory signals. Harnessing the apoptosis-inducing potential of Fas for therapy of cancer and autoimmune disease has been actively pursued, and despite a number of unexpected side-effects that result from manipulating Fas-FasL interactions, this remains a worthy goal. PMID- 19132325 TI - Cancer prevention knowledge of people with profound hearing loss. AB - BACKGROUND: Deaf persons, a documented minority population, have low reading levels and difficulty communicating with physicians. The effect of these on their knowledge of cancer prevention recommendations is unknown. METHODS: A cross sectional study of 222 d/Deaf persons in Michigan, age 18 and older, chose one of four ways (voice, video of a certified American Sign Language interpreter, captions, or printed English) to complete a self-administered computer video questionnaire about demographics, hearing loss, language history, health-care utilization, and health-care information sources, as well as family and social variables. Twelve questions tested their knowledge of cancer prevention recommendations. The outcome measures were the percentage of correct answers to the questions and the association of multiple variables with these responses. RESULTS: Participants averaged 22.9% correct answers with no gender difference. Univariate analysis revealed that smoking history, types of medical problems, last physician visit, and women having previous cancer preventive tests did not affect scores. Improved scores occurred with computer use (p = 0.05), higher education (p < 0.01) and income (p = 0.01), hearing spouses (p < 0.01), speaking English in multiple situations (p < 0.001), and in men with previous prostate cancer testing (p = 0.04). Obtaining health information from books (p = 0.05), physicians (p = 0.008), nurses (p = 0.03) or the internet (p = 0.02), and believing that smoking is bad (p < 0.001) also improved scores. Multivariate analysis revealed that English use (p = 0.01) and believing that smoking was bad (p = 0.05) were associated with improved scores. CONCLUSION: Persons with profound hearing loss have poor knowledge of recommended cancer prevention interventions. English use in multiple settings was strongly associated with increased knowledge. PMID- 19132327 TI - Anaerobic ethylene glycol degradation by microorganisms in poplar and willow rhizospheres. AB - Although aerobic degradation of ethylene glycol is well documented, only anaerobic biodegradation via methanogenesis or fermentation has been clearly shown. Enhanced ethylene glycol degradation has been demonstrated by microorganisms in the rhizosphere of shallow-rooted plants such as alfalfa and grasses where conditions may be aerobic, but has not been demonstrated in the deeper rhizosphere of poplar or willow trees where conditions are more likely to be anaerobic. This study evaluated ethylene glycol degradation under nitrate-, and sulphate-reducing conditions by microorganisms from the rhizosphere of poplar and willow trees planted in the path of a groundwater plume containing up to 1.9 mol l(-1) (120 g l(-1)) ethylene glycol and, the effect of fertilizer addition when nitrate or sulphate was provided as a terminal electron acceptor (TEA). Microorganisms in these rhizosphere soils degraded ethylene glycol using nitrate or sulphate as TEAs at close to the theoretical stoichiometric amounts required for mineralization. Although the added nitrate or sulphate was primarily used as TEA, TEAs naturally present in the soil or CO(2) produced from ethylene glycol degradation were also used, demonstrating multiple TEA usage. Anaerobic degradation produced acetaldehyde, less acetic acid, and more ethanol than under aerobic conditions. Although aerobic degradation rates were faster, close to 100% disappearance was eventually achieved anaerobically. Degradation rates under nitrate-reducing conditions were enhanced upon fertilizer addition to achieve rates similar to aerobic degradation with up to 19.3 mmol (1.20 g) of ethylene glycol degradation l(-1) day(-1) in poplar soils. This is the first study to demonstrate that microorganisms in the rhizosphere of deep rooted trees like willow and poplar can anaerobically degrade ethylene glycol. Since anaerobic biodegradation may significantly contribute to the phytoremediation of ethylene glycol in the deeper subsurface, the need for "pump and treat" or an aerobic treatment would be eliminated, hence reducing the cost of treatment. PMID- 19132326 TI - Effect of advanced access scheduling on processes and intermediate outcomes of diabetes care and utilization. AB - BACKGROUND: The impact of open access (OA) scheduling on chronic disease care and outcomes has not been studied. OBJECTIVE: To assess the effect of OA implementation at 1 year on: (1) diabetes care processes (testing for A1c, LDL, and urine microalbumin), (2) intermediate outcomes of diabetes care (SBP, A1c, and LDL level), and (3) health-care utilization (ED visits, hospitalization, and outpatient visits). METHODS: We used a retrospective cohort study design to compare process and outcomes for 4,060 continuously enrolled adult patients with diabetes from six OA clinics and six control clinics. Using a generalized linear model framework, data were modeled with linear regression for continuous, logistic regression for dichotomous, and Poisson regression for utilization outcomes. RESULTS: Patients in the OA clinics were older, with a higher percentage being African American (51% vs 34%) and on insulin. In multivariate analyses, for A1c testing, the odds ratio for African-American patients in OA clinics was 0.47 (CI: 0.29-0.77), compared to non-African Americans [OR 0.27 (CI: 0.21-0.36)]. For urine microablumin, the odds ratio for non-African Americans in OA clinics was 0.37 (CI: 0.17-0.81). At 1 year, in adjusted analyses, patients in OA clinics had significantly higher SBP (mean 6.4 mmHg, 95% CI 5.4 - 7.5). There were no differences by clinic type in any of the three health-care utilization outcomes. CONCLUSION: OA scheduling was associated with worse processes of care and SBP at 1 year. OA clinic scheduling should be examined more critically in larger systems of care, multiple health-care settings, and/or in a randomized controlled trial. PMID- 19132328 TI - Enrichment and identification of polycyclic aromatic compound-degrading bacteria enriched from sediment samples. AB - The degradation of polycyclic aromatic compounds (PACs) has been widely studied. Knowledge of the degradation of PACs by microbial populations can be utilized in the remediation of contaminated sites. To isolate and identify PAC-degrading bacteria for potential use in future bioremediation programmes, we established a series of PAC enrichments under the same experimental conditions from a single sediment sample taken from a highly polluted estuarine site. Enrichment cultures were established using the pollutants: anthracene, phenanthrene and dibenzothiophene as a sole carbon source. The shift in microbial community structure on each of these carbon sources was monitored by analysis of a time series of samples from each culture using 16S rRNA polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Significantly, our findings demonstrate that shifts in the constituent species within each degradative community are directly attributable to enrichment with different PACs. Subsequently, we characterized the microorganisms comprising the degradative communities within each enrichment using 16S rRNA sequence data. Our findings demonstrate that the ability to degrade PACs is present in five divisions of the Proteobacteria and Actinobacteria. By determining the precise identity of the PAC degrading bacterial species isolated from a single sediment sample, and by comparing our findings with previously published research, we demonstrate how bacteria with similar PAC degrading capabilities and 16S rRNA signatures are found in similarly polluted environments in geographically very distant locations, e.g., China, Italy, Japan and Hawaii. Such a finding suggests that geographical barriers do not limit the distribution of key PAC-degrading bacteria; this finding is in accordance with the Baas-Becking hypothesis "everything is everywhere; the environment selects" and may have significant consequences for the global distribution of PAC-degrading bacteria and their use in bioremediation. PMID- 19132329 TI - [Coughing attacks and reflux after extubation]. PMID- 19132330 TI - [Opioid-induced pruritus. Mechanisms and treatment regimens]. AB - Substantial progress has been achieved in recent years in research on the interaction between pain and pruritus. Over and above the known inhibition of pruritus by painful stimuli (e.g. scratching), a foundation for the explanation of opioid-induced pruritus was laid through the discovery of pruritus-specific neuronal processing channels. Although traditionally the degranulating effect of opioids on mast cells was assumed to be the essential mechanism, it is now clear that opioids can also induce itching at the spinal level. Neurons of the dorsal horn of the pain system inhibit spinal itch neurons. If this inhibition is weakened by opioids, the disinhibited itch neurons become active and mediate itching, without stimulation of the primary afferent peripheral nerves. Spinal triggering of itching is observed in particular by activation of mu-opioid receptors (mu-OR), while kappa-OR surprisingly suppress itch. The therapeutic implications of this interaction will be described. PMID- 19132331 TI - [Primary documentation quality for paper-assisted digital mission data documentation. Initial results from the air rescue service]. AB - BACKGROUND: With the assistance of digital pen and paper technology in the field of prehospital data reporting, it seems to be possible to fulfill the requirements of "documentation" as well as the requirements of "quality management". The aim of this study was to determine the "primary documentation quality" (PDQ) of a data reporting system based on digital pen and paper technology (so-called DINO) within a helicopter emergency medical service (HEMS) over a 6-month period. RESULTS: The PDQ is defined as the proportion of prehospital documented data, which is primarily recorded correctly by the data reporting system. For the national uniform core dataset (so-called MIND2) the PDQ was 96.7%, for "checkbox" and "numeric data fields" the PDQ was 99.8% and 93.6%, respectively and for "vital data" the PDQ was 96.7% [heart rate (HF), measurement of blood pressure] and 84.1% [peripheral oxygen saturation (S(p)O2), end tidal carbon dioxide concentration (etCO2), oxygen administration (O2)]. For "measurements" the PDQ was 96.9% (time stamps) and 86.9% (time frames), for "drugs" the PDQ was 43.6% (drug name) and 69.8% (drug dosage) and for"placement of infusion" 42% (infusion name) and 85.3% (infusion time), respectively. The average time for the "verification process" after mission completion was 1.5+/ 0.4 min/mission. CONCLUSIONS: The "primary documentation quality" of prehospital mission data reporting with the assistance of a digital pen and paper based documentation system (DINO) has been shown to completely fulfill the requirements of rapid and safe data documentation in actual missions, even at this early stage of development. PMID- 19132332 TI - [Progress in locating the genetic causes of schizophrenia]. AB - Elucidation of the pathogenesis of schizophrenia is progressing rapidly. The importance of the glutamatergic system and the glutamate receptor GRM3 were shown in both genetic and pharmacological studies of the new drug LY2140023. The zinc finger domain-containing gene ZNF804A could be identified as a new schizophrenia susceptibility gene, while large copy number variants at 1q21.1 and 15q13.3 now are seen as monogenic causes of schizophrenia. It is anticipated that the coming years will see further rapid progress in the unraveling of the causes of schizophrenia. PMID- 19132333 TI - The single Andigenum origin of Neo-Tuberosum potato materials is not supported by microsatellite and plastid marker analyses. AB - Neo-Tuberosum refers to cultivated potato adapted to long-day tuberization and a syndrome of related morphological and physiological traits, developed by intercrossing and selection of short-day adapted potatoes of the Solanum tuberosum Andigenum Group, native from the Andes of western Venezuela to northern Argentina. This re-creation of the modern potato helped support the theory of an Andigenum Group origin of potato in temperate regions and the possibility to access the largely untapped diversity of the Andigenum Group germplasm by base broadening breeding. This Neo-Tuberosum derived theory, the re-creation of the modern potato from Andigenum germplasm, has been universally accepted for almost 40 years, and has had tremendous impact in planning some breeding programs and supporting phylogenetic conclusions in cultivated potato. We show, with microsatellite (simple sequence repeat, SSR) and plastid DNA marker data, that Neo-Tuberosum germplasm is closely related to Chilotanum Group landraces from lowland south-central Chile rather than to Andigenum Group germplasm. We interpret this quite unexpected result to be caused by strong rapid selection against the original Andigenum clones after unintended hybridization with Chilotanum Group germplasm. In addition, we show that Neo-Tuberosum and Andigenum Group germplasm did not serve to broaden the overall genetic diversity of advanced potato varieties, but rather that Neo-Tuberosum lines and lines not using this germplasm are statistically identical with regard to genetic diversity as assessed by SSRs. These results question the long-standing Neo-Tuberosum derived theory and have implications in breeding programs and phylogenetic reconstructions of potato. PMID- 19132334 TI - Forward and reverse genetics of rapid-cycling Brassica oleracea. AB - Seeds of rapid-cycling Brassica oleracea were mutagenized with the chemical mutagen, ethylmethane sulfonate. The reverse genetics technique, TILLING, was used on a sample population of 1,000 plants, to determine the mutation profile. The spectrum and frequency of mutations induced by ethylmethane sulfonate was similar to that seen in other diploid species such as Arabidopsis thaliana. These data indicate that the mutagenesis was effective and demonstrate that TILLING represents an efficient reverse genetic technique in B. oleracea that will become more valuable as increasing genomic sequence data become available for this species. The extensive duplication in the B. oleracea genome is believed to result in the genetic redundancy that has been important for the evolution of morphological diversity seen in today's B. oleracea crops (broccoli, Brussels sprouts, cauliflower, cabbage, kale and kohlrabi). However, our forward genetic screens identified 120 mutants in which some aspect of development was affected. Some of these lines have been characterized genetically and in the majority of these, the mutant trait segregates as a recessive allele affecting a single locus. One dominant mutation (curly leaves) and one semi-dominant mutation (dwarf like) were also identified. Allelism tests of two groups of mutants (glossy and dwarf) revealed that for some loci, multiple independent alleles have been identified. These data indicate that, despite genetic redundancy, mutation of many individual loci in B. oleracea results in distinct phenotypes. PMID- 19132335 TI - Differentially expressed genes during malting and correlation with malting quality phenotypes in barley (Hordeum vulgare L.). AB - Breeding for malting quality is an important goal of malting barley breeding programs. Malting quality is a complex phenotype that combines a large number of interrelated components, each of which shows complex inheritance. Currently, only a few genes involved in determining malting quality have been characterized. We combined transcript profiling with phenotypic correlations to identify candidate genes for malting quality. The Barley1 GeneChip array containing 22,792 probe sets was used to conduct transcript profiling of genes expressed in several different stages of malting of four malting cultivars. Genes that were differentially expressed in comparisons between different malting stages relative to ungerminated seed, as well as in comparisons between malting cultivars in the same malting stage were identified. Correlation analysis of 723 differentially expressed genes with malting quality phenotypes showed that 11-102 of these genes correlated with six malting quality phenotypes. Genes involved in carbohydrate metabolism were among the positively correlated genes. Genes for protein and lipid metabolism, cell wall organization and biogenesis, and genes involved in stress and defense response also correlated with malting quality phenotypes. Expressed sequence tags (ESTs) were generated from a 'malting-gene enriched' cDNA library made by suppression subtractive hybridization between malted and ungerminated seeds of 'Morex'. Eleven percent of the ESTs had no significant homology with sequences in the databases, suggesting that there may be other malting-related genes not represented in the barley gene chip array. The results provide candidate genes for malting quality phenotypes that need to be functionally validated. PMID- 19132336 TI - A detailed synteny map of the eggplant genome based on conserved ortholog set II (COSII) markers. AB - We report herein the mapping of 115 PCR-based orthologous markers, including 110 conserved ortholog set or COSII markers, on the reference RFLP map of eggplant. The result permitted inference of a detailed syntenic relationship between the eggplant and tomato genomes. Further, the position of additional 522 COSII markers was inferred in the eggplant map via eggplant-tomato synteny, bringing the total number of markers in the eggplant genome to 869. Since divergence from their last common ancestor approximately 12 million years ago, the eggplant and tomato genomes have become differentiated by a minimum number of 24 inversions and 5 chromosomal translocations, as well as a number of single gene transpositions possibly triggered by transposable elements. Nevertheless, the two genomes share 37 conserved syntenic segments (CSSs) within which gene/marker order is well preserved. The high-resolution COSII synteny map described herein provides a platform for cross-reference of genetic and genomic information (including the tomato genome sequence) between eggplant and tomato and therefore will facilitate both applied and basic research in eggplant. PMID- 19132337 TI - Bayesian multilocus association mapping on ordinal and censored traits and its application to the analysis of genetic variation among Oryza sativa L. germplasms. AB - Association mapping can be a powerful tool for detecting quantitative trait loci (QTLs) without requiring line-crossing experiments. We previously proposed a Bayesian approach for simultaneously mapping multiple QTLs by a regression method that directly incorporates estimates of the population structure. In the present study, we extended our method to analyze ordinal and censored traits, since both types of traits are common in the evaluation of germplasm collections. Ordinal probit and tobit models were employed to analyze ordinal and censored traits, respectively. In both models, we postulated the existence of a latent continuous variable associated with the observable data, and we used a Markov-chain Monte Carlo algorithm to sample the latent variable and determine the model parameters. We evaluated the efficiency of our approach by using simulated- and real-trait analyses of a rice germplasm collection. Simulation analyses based on real marker data showed that our models could reduce both false-positive and false-negative rates in detecting QTLs to reasonable levels. Simulation analyses based on highly polymorphic marker data, which were generated by coalescent simulations, showed that our models could be applied to genotype data based on highly polymorphic marker systems, like simple sequence repeats. For the real traits, we analyzed heading date as a censored trait and amylose content and the shape of milled rice grains as ordinal traits. We found significant markers that may be linked to previously reported QTLs. Our approach will be useful for whole-genome association mapping of ordinal and censored traits in rice germplasm collections. PMID- 19132340 TI - Prolonging resuscitation and postponing the death of activated protein C or is it? PMID- 19132338 TI - Pilot safety study of low-dose vasopressin in non-septic critically ill children. AB - OBJECTIVE: To assess the safety of low-dose vasopressin infusion in critically ill children requiring prolonged mechanical ventilation (MV) at risk of developing sedation/analgesia-related hypotension. METHOD: Randomized pilot safety study in children expected to require MV for at least 3 days. Children received either vasopressin (0.0005 U/kg/min) or sodium chloride (0.9%) infusion for a period of 48 h. Haemodynamic variables, urine output and serum electrolytes were closely monitored and analyzed. RESULTS: Twelve children in each group had similar baseline characteristics. Vasopressin infusion was associated with an 8 mmol/L fall in serum sodium concentration (p < 0.01) and with higher incidence of hyponatraemia (8 vs. 66%, p < 0.01). In normotensive children, low-dose vasopressin also induced a reversible decrease in urine output, and acutely increased blood pressure (p < 0.01). After stopping the vasopressin there was rebound hypotension (p < 0.01). CONCLUSION: Low-dose vasopressin infusion in haemodynamically stable, but critically ill, children is associated with reduction in urine output and decreased serum sodium level, yielding a high incidence of hyponatraemia. We conclude that these effects limit further study of prophylactic vasopressin for sedation-related hypotension in a randomized controlled trial. PMID- 19132342 TI - Comment on "Indwelling time and risk of colonization of peripheral arterial catheters in critically ill patients" by Khalifa et al. PMID- 19132343 TI - Mini-series: I. Basic science. Uncertainty and inaccuracy of predicting CYP mediated in vivo drug interactions in the ICU from in vitro models: focus on CYP3A4. AB - Drug-drug interactions (DDIs) contribute significantly to the incidence of adverse drug reactions. Important advances in the knowledge of human drug metabolizing enzymes have fueled the integration of in vitro drug metabolism and clinical DDIs studies for use in drug development programs and in the clinical setting. The activity of cytochrome P450 (CYP) 3A4 and P-glycoprotein are critical determinant of drug clearance, interindividual variability in drug disposition and clinical efficacy, and appears to be involved in the mechanism of numerous clinically relevant DDIs. Cell-based in vitro models are being increasingly applied in elucidating the pharmacokinetic profile of drug candidates during the preclinical steps of drug development. Human liver, intestinal samples and recombinant human CYP3A4 are now readily available as in vitro screening tools to predict the potential for in vivo DDIs. Although it is easy to determine in vitro metabolic DDIs, the interpretation and extrapolation of in vitro interaction data to in vivo situations requires a good understanding of pharmacokinetic principles. Clinicians and pharmacokineticists should recognize that in vitro models may not be clinically relevant in all situations. In the current article, research will be presented on drug metabolism and DDIs along with examples illustrating the utility of specific in vitro or in vivo approaches. In addition, the impact and clinical relevance of complexities such as dosing-route dependent effects, multi-site kinetics of drug-metabolizing enzymes and non-CYP determinants of metabolic clearance will be addressed. PMID- 19132344 TI - Mini-series: II. clinical aspects. clinically relevant CYP450-mediated drug interactions in the ICU. AB - BACKGROUND: In the critically ill, multiple drug therapies for acute and chronic conditions are often used at the same time and adverse drug events occur frequently. Many pharmacological and disease-related factors, e.g. altered renal and hepatic function, catecholamine-related circulatory changes, altered drug volume of distribution, enteral versus parenteral feeding and morbid obesity, along with concomitant multiple drug regimens may account for the wide inter individual variability in drug exposure and response in critically ill patients and for the high risk for drug-drug interactions to occur. The practicing intensivist must remain aware of the major mechanisms for drug-drug interactions, among which the drug-metabolizing enzyme inhibitory or induction potential of associated chemical entities are paramount. Metabolism-based drug-drug interactions are largely due to changes in levels of drug-metabolizing enzymes caused by one drug, leading to changes in the systemic exposure clearance of another. Among the numerous drug-metabolizing enzymes identified to date, the activity of cytochrome P450s (CYP450) is a critical determinant of drug clearance and appears to be involved in the mechanism of numerous clinically relevant drug drug interactions observed in critically ill patients. DISCUSSION: This manuscript will cover a practical overview of clinically relevant CYP450-mediated drug-drug interactions. Medications frequently used in the intensive care unit such as benzodiazepines, immunosuppressive agents, opioid analgesics, certain anticonvulsants, the azoles and macrolides have the potential to interact with CYP450-mediated metabolism and may lead to toxicity or therapeutic failure. PMID- 19132345 TI - Accuracy of side-cutting implantation instruments for total knee arthroplasty. AB - A new generation of implantation instruments were developed for quadriceps sparing surgical approaches during total knee arthroplasty (TKA). There is little information on the accuracy of the bone cuts performed with the side-cutting technique. A total of 100 patients were randomized to undergo computer-assisted TKA or non-navigated TKA using a mini-subvastus surgical approach and side cutting implant instrumentation. The radiographic parameters, clinical outcomes and knee scores were evaluated 3 months postoperative. The mechanical axis of the limb was within 3 degrees varus/valgus in 76% of the patients who had navigated procedures versus 66% of patients who had conventional surgery. The tibial slope showed a rate of inaccuracy of 3 degrees or less for 78% of the patients in the navigated total knee arthroplasty group versus 66% of the patients in the conventional group. Clinical outcomes and knee scores were similar in both groups. The navigation technique could not compensate for shortcomings of the implantation instruments. PMID- 19132346 TI - Epidemiologic, clinical and arthroscopic study of the discoid meniscus variant in Greek population. AB - This retrospective study was aimed to investigate the epidemiologic, clinical and arthroscopic features of discoid meniscus variant in Greek population. We reviewed the cases of 2,132 patients who underwent knee arthroscopy between 1986 and 2004 and diagnosis of discoid lateral meniscus was established in 39 patients with mean age of 31.7 +/- 9.4 years old. Incidence of the discoid lateral meniscus variant was recorded at rate of 1.8% presenting no significant differences according to patient gender or lesion body side. Regarding the type of discoid dysmorphy, 23 cases attributed to complete type, 15 were incomplete and in one case, Wrisberg type was observed. Predictive values of the most commonly recorded physical signs in the clinical diagnosis of the discoid meniscus were analysed. Comparative evaluation of the long-term results of arthroscopic partial meniscectomy performed in patients with intact or torn discoid lateral meniscus and torn normally shaped lateral meniscus was carried out using Lysholm and IKDC scoring systems. Also, we investigated any correlation between dysmorphy type and tear pattern analysing the arthroscopic findings. Results demonstrated that the discoid meniscus lesion represents an atypical clinical entity in adults and no significant predictive value of the signs encountered in the clinical examination of the patients with discoid meniscus was observed. Clinical outcome after arthroscopic partial meniscectomy regarding the intact discoid meniscus group was superior in comparison with that of torn discoid meniscus cases. On other hand, no difference in the result of partial meniscectomy between discoid and normal lateral meniscus tear groups was found. No statistically significant relationship between the type of discoid menisci and tear pattern or incidence rate of concomitant intraarticular lesions was confirmed. PMID- 19132347 TI - Management of overtight medial patellofemoral ligament reconstruction. AB - The clinical presentation of an overtight medial patellofemoral ligament (MPFL) reconstruction can differ depending on whether it is too tight in extension (extensor lag) or too tight in flexion (anterior knee pain and loss of flexion). We report one clinical case of each presentation. Both cases were treated with a percutaneous release of the graft. After the release, both patients regained a full active range of motion without residual symptoms. These complications demonstrate that the adjustment of the graft tensioning as well as its femoral position are critical steps in MPFL reconstruction. This procedure requires training and experience in order to avoid early complications related to malposition or inappropriate tensioning of the graft. A surgical management for these overtight reconstructions is recommended, as it will restore function and range of motion, and prevent late patellofemoral degeneration. PMID- 19132350 TI - Post-operative analysis of ACL tibial fixation. AB - The main aims of the study were to describe the mechanical behaviour of the bone and tendon graft in ACL reconstruction. We also wanted to evaluate the stresses upon the tendon and try to relate these to modes of healing observed in animal studies. An MRI scan of a knee from a 22-year-old male patient, 3 months post ACL reconstruction, was used to generate a finite element model for assessing the stresses on the tendon and bone. A 200 N force, equivalent to the force during normal gait, was placed on the graft and the stress distribution recorded. The maximum stress was found on the tendon graft at the proximal end of the tibial tunnel. Direct tendon healing, which has been observed in animal studies, maybe as a result of the increased stress in this area. PMID- 19132348 TI - Functional outcomes and health-related quality of life after robot-assisted anterior cruciate ligament reconstruction with patellar tendon grafts. AB - During a short period of time, surgical robots had been propagated for automated tunnel placement in anterior cruciate ligament (ACL) reconstruction. Clinical outcome data are currently unavailable. Between 2000 and 2003, 152 patients underwent ACL replacement with the assistance of the Computer Assisted Surgical Planning and Robotics system (CASPAR, OrtoMaquet, Germany) at our hospital. After minimal invasive pin placement in both the tibia and femur, computed tomography was used to register anatomical landmarks and to plan graft tunnel alignment. The robot was used to drill tibial and femoral tunnels in an outside-in fashion according to pre-operative planning. There was one procedure-specific Serious Adverse Event (i.e., an intraoperative transection of the posterior cruciate ligament). After IRB approval, all patients were invited for a follow-up examination. Data from 100 patients (35 women, 65 men, mean age 35 [SD 11] years, median follow-up 61 [range 42-77] months) form the basis of this report. Side-to side differences in anterior laxity were measured with the KT-1000 arthrometer. Patient-centered outcomes included the Lysholm-Score, the lower extremity functional scale (LEFS), and the Short Form 36 (SF36). The mean KT-1000 side-to side difference was 0.89 [95% confidence interval (CI) 0.52-1.26] mm. Eight and five patients had a positive Lachman and pivot shift test, respectively. The Lysholm-Score averaged 86 (95% CI 83-89) points. Excellent, good, fair, and poor outcomes were reported by 38, 32, 20, and 10 patients. The LEFS averaged 85 (95% CI 82-88) points. The mean SF36 Physical Component Score was 48.4 (95% CI 46.5 50.3), indicating residual deficits compared to the population norm. All tibial graft tunnels did not cross the Blumensaat line, but were placed slightly anterior to the optimal center of 42% reported in previous studies. Compared to literature data, robot-assisted ACL reconstruction with BTB grafts may lead to higher knee stability, but poorer functional outcomes. The immense additional efforts with the procedure did not pay off in a benefit to patients. PMID- 19132352 TI - Evaluation of gas chromatography columns for the analysis of the 15 + 1 EU priority polycyclic aromatic hydrocarbons (PAHs). AB - Three different stationary phases were investigated for the analysis of the 15 + 1 EU-priority polycyclic aromatic hydrocarbons (PAHs) by gas chromatography-mass spectrometry. In addition to the most commonly used 5% phenyl methylpolysiloxane, a mid-polar phase (50% phenyl methylpolysiloxane) and a recently commercialised mid-polar to polar phase (Optima delta-6), were evaluated. Challenging groups of PAHs in terms of separation, such as the pair dibenz[a,h]anthracene-indeno[1,2,3, cd]pyrene and the two groups benzo[b]fluoranthene-benzo[k]fluoranthene benzo[j]fluoranthene and cyclopenta[cd]pyrene-benz[a]anthracene-chrysene, were satisfactorily separated by using the mid-polar phase. Moreover, discrimination in terms of peak height for the heaviest PAHs (caused from the strong interaction of these compounds with the stationary phase) was reduced without compromising the resolution of the other target analytes when applying the mid-polar phase in a tailor-made column geometry (20 m x 0.18 mm internal diameter and 0.14 microm film thickness) in combination with optimised chromatographic conditions. A significant enhancement of the analytical sensitivity for dibenzopyrenes is demonstrated with an almost threefold increase of the signal-to-noise (S/N) ratio for dibenzo[a,h]pyrene, the last eluting PAH. The ability of the selected column to separate potentially interfering PAHs from the target analytes in both solvent solutions and food extracts is demonstrated. PMID- 19132351 TI - The adenosine A2A antagonist MSX-3 reverses the effects of the dopamine antagonist haloperidol on effort-related decision making in a T-maze cost/benefit procedure. AB - RATIONALE: Mesolimbic dopamine (DA) is a critical component of the brain circuitry regulating behavioral activation and effort-related processes. Research involving choice tasks has shown that rats with impaired DA transmission reallocate their instrumental behavior away from food-reinforced tasks with high response requirements and instead select less effortful food-seeking behaviors. OBJECTIVE: Previous work showed that adenosine A(2A) antagonism can reverse the effects of the DA antagonist haloperidol in an operant task that assesses effort related choice. The present work used a T-maze choice procedure to assess the effects of adenosine A(2A) and A(1) antagonism. MATERIALS AND METHODS: With this task, the two arms of the maze have different reinforcement densities (four vs. two food pellets), and a vertical 44 cm barrier is positioned in the arm with the higher density, presenting the animal with an effort-related challenge. Untreated rats strongly prefer the arm with the high density of food reward and climb the barrier in order to obtain the food. RESULTS: Haloperidol produced a dose-related (0.05-0.15 mg/kg i.p.) reduction in the number of trials in which the rats chose the high-barrier arm. Co-administration of the adenosine A(2A) receptor antagonist MSX-3 (0.75, 1.5, and 3.0 mg/kg i.p.), but not the A(1) antagonist 8 cyclopentyl-1,3-dipropylxanthine (0.75, 1.5, and 3.0 mg/kg i.p.), reversed the effects of haloperidol on effort-related choice and latency. CONCLUSIONS: Adenosine A(2A) and D2 receptors interact to regulate effort-related decision making, which may have implications for the treatment of psychiatric symptoms such as psychomotor slowing or anergia that can be observed in depression, parkinsonism, and other disorders. PMID- 19132353 TI - A MIP-based flow-through fluoroimmunosensor as an alternative to immunosensors for the determination of digoxin in serum samples. AB - This work reports a comparative study of two automated flow-through fluorosensors for the determination of digoxin in serum samples: an immunosensor with an anti digoxin polyclonal antibody as the reactive phase permanently immobilised on controlled-pore glass and a sensor with a selective reaction system based on a methacrylic molecularly imprinted polymer (MIP) synthesised by bulk polymerisation. The variables affecting the sensitivity and dynamic range of the sensors (e.g. the carrier and elution solutions, flow rates, pH and reagent concentrations) were optimized, and the binding characteristics of their reactive phases were compared in a competitive fluorescent assay. Digoxin was reproducibly determined by both sensors at the milligram per litre level (detection limit = 1.20 x 10(-3) mg L(-1) and RSD = 4-7% for the immunosensor; detection limit = 1.7 x 10(-5) mg L(-1) and RSD = 1-2% for the MIP sensor). No cross-reactivity with digoxin-related compounds was seen for either sensor at a digoxin/interferent ratio of 1:100. The lifetime of the immunosensor was about 50 immunoassays; its shelf life, when unused, is about 3 months. The lifetime of the MIP sensor was over 18 months. Both sensors were used to determine the digoxin concentration of human serum samples with satisfactory results. PMID- 19132354 TI - MicroRNA detection by microarray. AB - MicroRNAs (miRNAs) are a class of small noncoding RNAs approximately 22 nt in length that regulate gene expression and play fundamental roles in multiple biological processes, including cell differentiation, proliferation and apoptosis as well as disease processes. The study of miRNA has thus become a rapidly emerging field in life science. The detection of miRNA expression is a very important first step in miRNA exploration. Several methodologies, including cloning, northern blotting, real-time RT-PCR, microRNA arrays and ISH (in situ hybridization), have been developed and applied successfully in miRNA profiling. This review discusses the main existing microRNA detection technologies, while emphasizing microRNA arrays. PMID- 19132355 TI - Diffusion tensor imaging: the normal evolution of ADC, RA, FA, and eigenvalues studied in multiple anatomical regions of the brain. AB - INTRODUCTION: The aim of our work was to investigate the process of myelination in healthy patients using the diffusion parameters apparent diffusion coefficient (ADC), relative anisotropy (RA), fractional anisotropy (FA), and eigenvalues. Age dependent changes were assessed using the slope m of the fit functions that best described the data. MATERIALS AND METHODS: Seventy-two patients (3 weeks-19 years) without pathological magnetic resonance imaging findings were selected from all pediatric patients scanned with diffusion tensor imaging over a 5-year period at our institution. ADC, RA, FA, and eigenvalue maps were calculated and regions of interest were selected in anterior/posterior pons, genu/splenium of corpus callosum (CC), anterior/posterior limb of internal capsule (IC), and white matter (WM) regions (frontal, temporal, parietal, occipital WM). Statistical analysis was performed using Spearman correlation coefficient and regression analysis. RESULTS: Mean values ranged 71.6 x 10(-5) to 90.3 x 10(-5) mm(2)/s (pons/parietal WM) for ADC, 0.32-0.94 (frontal WM/CC) for RA, and 0.36-0.81 (frontal WM/splenium) for FA. Logarithmic fit functions best described the data. Strong age influences were observed for CC, pons, and parietal/frontal WM and changes were significant for all three eigenvalues, most pronounced for perpendicular eigenvalues. Changes in RA and FA differed depending on the structure anisotropy. CONCLUSIONS: Changes observed for ADC, RA, FA, and eigenvalues with age were consistent with previous findings. Changes detected for RA and FA varied due to the different scaling of both parameters. We found that the use of the largely linear scaled RA adds more valuable information for the assessment of age-dependent structural changes as compared to FA. Additionally, we report normative values for the diffusion parameters studied. PMID- 19132356 TI - Preoperative use of ultrasonography to localize an ingested foreign body. AB - We report a 9-year-old boy who had swallowed a small dressmaker's pin during an art & craft class. Confirmation of ingestion of the pin and its passage through the gut was achieved with abdominal radiography. When the pin had not passed after 8 days, and with increasing concern about the likelihood of perforation, US was used to locate its exact position to allow surgical removal. This case report illustrates the unique use of US to reveal the intraappendiceal location of an ingested foreign body, facilitating its surgical removal. PMID- 19132357 TI - Congenital persistent terminal ventricle and filar cyst. PMID- 19132358 TI - Gastric pneumatosis and portal venous gas: benign findings in hypertrophic pyloric stenosis. PMID- 19132359 TI - Mechanical model for a collagen fibril pair in extracellular matrix. AB - In this paper, we model the mechanics of a collagen pair in the connective tissue extracellular matrix that exists in abundance throughout animals, including the human body. This connective tissue comprises repeated units of two main structures, namely collagens as well as axial, parallel and regular anionic glycosaminoglycan between collagens. The collagen fibril can be modeled by Hooke's law whereas anionic glycosaminoglycan behaves more like a rubber-band rod and as such can be better modeled by the worm-like chain model. While both computer simulations and continuum mechanics models have been investigated for the behavior of this connective tissue typically, authors either assume a simple form of the molecular potential energy or entirely ignore the microscopic structure of the connective tissue. Here, we apply basic physical methodologies and simple applied mathematical modeling techniques to describe the collagen pair quantitatively. We found that the growth of fibrils was intimately related to the maximum length of the anionic glycosaminoglycan and the relative displacement of two adjacent fibrils, which in return was closely related to the effectiveness of anionic glycosaminoglycan in transmitting forces between fibrils. These reveal the importance of the anionic glycosaminoglycan in maintaining the structural shape of the connective tissue extracellular matrix and eventually the shape modulus of human tissues. We also found that some macroscopic properties, like the maximum molecular energy and the breaking fraction of the collagen, were also related to the microscopic characteristics of the anionic glycosaminoglycan. PMID- 19132360 TI - Modeling tumor cell shedding. AB - Cell shedding is an important step in the development of tumor invasion and metastasis. It influences growth saturation, latency, and tumor surface roughness. In spite of careful experiments carried out using multicellular tumor spheroids (MTS), the effects of the shedding process are not yet completely understood. Using a simulational model, we study how the nature and intensity of cell shedding may influence tumor morphology and examine the dependence of the total number of shed cells with the relevant parameters, finding the ranges that maximize cell detachment. These ranges correspond to intermediate values of the adhesion, for which we observe the emergence of a rough tumor surface. They are also likely to maximize the probability of generating invasion and metastases. Using numerical values taken from experiments, we find that the shedding-induced reduction in the growth rate is not intense enough to lead to latency, except when cell adhesion is assumed to be very weak. This suggests that the presence of inhibitors is a necessary condition for the observed MTS growth saturation. PMID- 19132362 TI - Macrophage ion currents are fit by a fractional model and therefore are a time series with memory. AB - We studied macroscopic ion currents from macrophages and compared their patterns of behavior using classical and fractal analysis. Peak and steady state currents were measured respectively at the beginning and end of a voltage-clamp pulse. Hurst coefficients H and fractional dimensions were calculated for the current fluctuations (I(H)) during the intervening interval; these fluctuations are usually assumed to be white noise. We show that I(H) is different from 0.5 and that the increments are stationary, indicating that the dynamic model has memory and that the intervening current fluctuations cannot be considered as white noise. I(H) is less than 0.5, implying an antipersistent pattern. In addition, we show that the relation between inactivation and I(H) versus voltage V fit an equation I(H)(V) = f(V, alpha, m, d), where alpha is associated with fractional calculus and m and d are free parameters. Fitting by a fractional model confirms that the phenomenon has memory. PMID- 19132361 TI - Movements of native C505 during channel gating in CNGA1 channels. AB - We investigated conformational changes occurring in the C-linker and cyclic nucleotide-binding (CNB) domain of CNGA1 channels by analyzing the inhibition induced by thiol-specific reagents in mutant channels Q409C and A414C in the open and closed state. Cd(2+) (200 microM) inhibited irreversibly mutant channels Q409C and A414C in the closed but not in the open state. Cd(2+) inhibition was abolished in the mutant A414C(cys-free), in the double mutant A414C + C505T and in the tandem construct A414C + C505T/CNGA1, but it was present in the construct A414C + C505(cys-free). The cross-linker reagent M-2-M inhibited mutant channel Q409C in the open state. M-2-M inhibition in the open state was abolished in the double mutant Q409C + C505T and in the tandem construct Q409C + C505T/CNGA1. These results show that C(alpha) of C505 in the closed state is located at a distance between 4 and 10.5 A from the C(alpha) of A414 of the same subunit, but in the open state C505 moves towards Q409 of the same subunit at a distance that ranges from 10.5 to 12.3 A from C(alpha) of this residue. These results are not consistent with a 3-D structure of the CNGA1 channel homologous to the structure of HCN2 channels either in the open or in the closed state. PMID- 19132363 TI - The role of protons in fast and slow gating of the Torpedo chloride channel ClC 0. AB - Transmembrane proton transport is of fundamental importance for life. The list of H(+) transporting proteins has been recently expanded with the discovery that some members of the CLC gene family are stoichiometrically coupled Cl(-)/H(+) antiporters. Other CLC proteins are instead passive Cl(-) selective anion channels. The gating of these CLC channels is, however, strongly regulated by pH, likely reflecting the evolutionary relationship with CLC Cl(-)/H(+) antiporters. The role of protons in the gating of the model Torpedo channel ClC-0 is best understood. ClC-0 is a homodimer with separate pores in each subunit. Each protopore can be opened and closed independently from the other pore by a "fast gate". A common, slow gate acts on both pores simultaneously. The opening of the fast gate is controlled by a critical glutamate (E166), whose protonation state determines the fast gate's pH dependence. Extracellular protons likely can arrive directly at E166. In contrast, protonation of E166 from the inside has been proposed to be mediated by the dissociation of an intrapore water molecule. The OH(-) anion resulting from the water dissociation is stabilized in one of the anion binding sites of the channel, competing with intracellular Cl(-) ions. The pH dependence of the slow gate is less well understood. It has been shown that proton translocation drives irreversible gating transitions associated with the slow gate. However, the relationship of the fast gate's pH dependence on the proton translocation and the molecular basis of the slow gate remain to be discovered. PMID- 19132364 TI - Effect of headgroup on the dipole potential of phospholipid vesicles. AB - The dipole potentials, psi (d), of phospholipid vesicles composed of pure dimyristoylphosphatidylcholine (DMPC) or vesicles in which 50 mol% of the DMPC was substituted by dimyristoylphosphatidylserine (DMPS), dimyristoylphosphatidylglycerol (DMPG), dimyristoylethanolamine (DMPE), dimyristoylphosphatidic acid (DMPA) or monomyristoylphosphatidylcholine (MMPC) were measured via a fluorescent ratiometric method utilizing the probe di-8 ANEPPS. The PS and PG headgroups were found to cause only minor changes in psi (d). PE caused an increase in psi (d) of 51 mV. This could be explained by a decrease in the dielectric constant of the glycerol backbone region as well as a movement of the P(-)-N(+) dipole of the less bulky PE headgroup to a position more parallel to the membrane surface than in PC. The negatively charged PA headgroup increases psi (d) by 215 mV relative to PC alone. This indicates that the positive pole of the dipole predominantly responsible for the dipole potential is located at a position closer to the interior of the membrane than the phosphate group. The increase in the charge of the negative pole of the dipole by the phosphate group of PA increases the electrical potential drop across the lipid headgroup region. The incorporation of the single chain lipid MMPC into the membrane causes a decrease in psi (d) of 142 mV. This can be explained by a decrease in packing density within the membrane of carbonyl dipoles from the sn-2 chain of DMPC. The results presented should contribute to a better understanding of the electrical effect of lipid headgroups on the functioning of membrane proteins. PMID- 19132365 TI - Stability of microbubbles prepared by co-axial electrohydrodynamic atomisation. AB - Previous studies have indicated that microbubbles prepared by co-axial electrohydrodynamic atomisation (CEHDA) are less stable than those prepared by other methods such as sonication and microfluidic techniques. The aim of this investigation was to determine the reasons for this observation and how this might be addressed in future work. Microbubbles were prepared by CEHDA using (i) a glycerol-air system, (ii) a glycerol-Tween 80-air system and (iii) a glycerol zirconia-air system and also by simple agitation of (i) and (ii), in order to compare the effect upon the dissolution rate of microbubbles of different materials and processing methods. Both theoretical examination and the experimental results indicated that all three quantities were important in controlling the rate of microbubble dissolution, namely: surface tension at the gas/liquid interface, the effective diffusivity of gas through this interface and the initial concentration of gas dissolved in the surrounding liquid. However, it was the difference in gas concentration in the surrounding liquid that was indicated as the primary reason for the differences in stability observed with different processing methods. It was concluded, therefore, that improved stability could be achieved for microbubbles prepared using CEHDA by saturating the collecting fluid with gas and/or maintaining a high concentration of microbubbles during collection. PMID- 19132367 TI - Thermal aggregation and ion-induced cold-gelation of bovine serum albumin. AB - Protein cold-gelation has recently received particular attention for its relevance in bio and food technology. In this work, we report a study on bovine serum albumin cold-gelation induced by copper or zinc ions. Metal-induced cold gelation of proteins requires two steps: during the first one, the heat treatment causes protein partial unfolding and aggregation; then, after cooling the solution to room temperature, gels are formed upon the addition of metal ions. The thermally induced behaviour has been mainly investigated through different techniques: Fourier transform infrared (FTIR) spectroscopy, circular dichroism, dynamic light scattering (DLS) and rheology. Data have shown that the aggregation process is mainly due to protein conformational changes--alpha-helices into beta aggregates-forming small aggregated structures with a mean diameter of about 20 nm a few minutes after heating. After metal ion addition, the viscoelastic properties of the gels have been investigated by rheological measurements. The behaviour of the elastic and viscous moduli as a function of time is discussed in terms of ion concentration and type. Our results show that: (1) the elastic behaviour depends on ion concentration and (2) at a given ion concentration, gels obtained in the presence of zinc exhibit an elastic value larger than that observed in the Cu(2+) case. Data suggest that cold-gelation is the result of different mechanisms: the ion-mediated protein-protein interaction and the bridging effect due to the presence of divalent ions in solution. PMID- 19132366 TI - FLIM and emission spectral analysis of caspase-3 activation inside single living cell during anticancer drug-induced cell death. AB - Two-photon excitation (TPE) fluorescence lifetime imaging microscopy (FLIM) and emission spectral imaging (ESI) are powerful tools for fluorescence resonance energy transfer (FRET) measurement. In this study, we use these two techniques to analyze caspase-3 activation inside single living cells during anticancer drug induced human lung adenocarcinoma (ASTC-a-1) cell death. TPE-ESI of SCAT3, a caspase-3 indicator based on FRET, was performed inside single living cell stably expressing SCAT3. The TPE-ESI measurement was performed using 780 nm excitation which was considered to selectively excite the donor ECFP of SCAT3 by measuring the emission ratio of 526 to 476 nm. The emission peak at 526 nm disappeared and that of 476 nm increased after STS or bufalin treatment, but taxol treatment did not induce a significant change for the SCAT3 emission spectra, indicating that caspase-3 was activated during STS- or bufalin-induced cell apoptosis, but was not involved in taxol-induced PCD. Fluorescence lifetime of ECFP inside living cells was acquired using FLIM. The lifetime of ECFP was the same as that of the control group after taxol treatment, but increased from 1.83 +/- 0.02 to 2.05 +/- 0.03 and 1.90 +/- 0.03 ns, respectively after STS and bufalin treatment, which agree with the results obtained using TPE-ESI. Taken together, TPE-FLIM and ESI analysis were proved to be valuable approaches for monitoring caspase-3 activation inside single living cells. PMID- 19132368 TI - Concentration dependent effect of GsMTx4 on mechanosensitive channels of small conductance in E. coli spheroplasts. AB - The spider peptide GsMTx4, at saturating concentration of 5 muM, is an effective and specific inhibitor for stretch-activated mechanosensitive (MS) channels found in a variety of eukaryotic cells. Although the structure of the peptide has been solved, the mode of action remains to be determined. Because of its amphipathic structure, the peptide is proposed to interact with lipids at the boundaries of the MS channel proteins. In addition, GsMTx4 has antimicrobial effects, inhibiting growth of several species of bacteria in the range of 5-64 microM. Previous studies on prokaryotic MS channels, which serve as model systems to explore the principle of MS channel gating, have shown that various amphipathic compounds acting at the protein-lipid interface affect MS channel gating. We have therefore analyzed the effect of different concentrations of extracellular GsMTx4 on MS channels of small conductance, MscS and MscK, in the cytoplasmic membrane of wild-type E. coli spheroplasts using the patch-clamp technique. Our study shows that the peptide GsMTx4 exhibits a biphasic response in which peptide concentration determines inhibition or potentiation of activity in prokaryotic MS channels. At low peptide concentrations of 2 and 4 microM the gating of the prokaryotic MS channels was hampered, manifested by a decrease in pressure sensitivity. In contrast, application of peptide at concentrations of 12 and 20 microM facilitated prokaryotic MS channel opening by increasing the pressure sensitivity. PMID- 19132369 TI - Characterization of beta-1,3-galactosyl-N-acetylhexosamine phosphorylase from Propionibacterium acnes. AB - Homologs of the beta-1,3-galactosyl-N-acetylhexosamine phosphorylase (GalHexNAcP) gene (gnpA) were cloned from the genomic DNA of Propionibacterium acnes JCM6425 and P. acnes JCM6473, showing 99.9% and 97.9% nucleotide sequence identity, respectively, with the ppa0083 gene from the genome-sequenced P. acnes KPA171202. No gnpA gene was detected in the genomic DNA of type strain P. acnes ATCC25746. The recombinant enzyme from P. acnes JCM6425 (GnpA) showed approximately 70 times higher specific activity of phosphorolysis on galacto-N-biose (Galbeta1- >3GalNAc, GNB) than that on lacto-N-biose I (Galbeta1-->3GlcNAc). K (m) value for GnpA on GNB was high, but GnpA did not exhibit activity on any derivatives of GNB examined. These results indicate that GnpA is GalHexNAcP which should be classified as galacto-N-biose phosphorylase. The large k (cat) value of GnpA on GalNAc suggests that GnpA would be a useful catalyst for the synthesis of GNB. PMID- 19132370 TI - MR-plastination-arthrography: a new technique used to study the distal tibiofibular syndesmosis. AB - PURPOSE: The purpose of this study was to describe a new technique called MR plastination arthrography to study both intra- and extra-articular anatomy. MATERIALS AND METHODS: In six human cadaveric lower legs MR arthrography was performed in either a one-step or two-step procedure. In the former a mixture of diluted Gadolinium and dyed polymer was injected. In the latter the dyed polymer was injected after arthrography wih diluted Gadolinium. Three-millimeter slices of these legs, obtained in a plane identical to that of the MR images, were plastinated according to the E12 technique of von Hagens. The plastination slices were subsequently compared with the MR images. RESULTS: The one-step procedure resulted in an inhomogeneous arthrogram. The two-step procedure resulted in a good correlation between the high-resolution MR images and plastination slices, as expressed by a good comparison of anatomic detail of the small syndesmotic recess. CONCLUSIONS: Images of the distal tibiofibular syndesmosis obtained with plastination arthrography correlated well with images acquired by MR arthrography when performed in a two-step procedure. PMID- 19132371 TI - Median nerve cross-sectional area and MRI diffusion characteristics: normative values at the carpal tunnel. AB - OBJECTIVE: Enlargement of the median nerve is an objective potential imaging sign of carpal tunnel syndrome. Diffusion tensor MRI (DTI) may provide additional structural information that may prove useful in characterizing median neuropathy. This study further examines normal values for median nerve cross-sectional area (CSA), apparent diffusion coefficient (ADC), and fractional anisotropy (FA). MATERIALS AND METHODS: Twenty-three wrists in 17 healthy volunteers underwent MRI of the wrist at 3 T. In 13 subjects, DTI was performed at a B value of 600 mm(2)/s. Median nerve CSA, ADC, and FA were analyzed at standardized anatomic levels. RESULTS: Mean (SD) median nerve CSA within the proximal carpal tunnel was 10.0 (3.4) mm(2). The mean (SD) FA of the median nerve was 0.71 (0.06) and 0.70 (0.13) proximal to and within the carpal tunnel, respectively. There was a significant difference between nerve CSA and ADC, but not FA, at the distal forearm and proximal carpal tunnel. Nerve CSA, ADC, and FA did not differ between men and women or between dominant and non-dominant wrists. Nerve CSA at the proximal carpal tunnel was positively correlated with subject age and body mass index. CONCLUSION: Our results suggest a 90% upper confidence limit for normal median nerve CSA of 14.4 mm(2) at the proximal carpal tunnel, higher than normal limits reported by many ultrasound studies. We observed a difference between the CSA and ADC, but not the FA, of the median nerve at the distal forearm and proximal carpal tunnel levels. PMID- 19132372 TI - Oxalosis in primary hyperoxaluria in infancy : Report of a case in a 3-month-old baby. Follow-up for 3 years and review of literature. AB - Primary hyperoxaluria (PH1) is a rare inborn autosomal recessive metabolic disorder due to the deficiency of hepatic alanine-glyoxylate-aminotransferase. This deficiency results in excessive synthesis and urinary excretion of oxalate, inducing renal stone formation and deposition of calcium oxalate in the kidney, bone, myocardium, and vessels (systemic oxalosis, SO) in the most severely affected individuals. We report renal and skeletal changes in a 3-month-old girl with PH1 and SO. Intense cortico-medullary hyperechogenicity and increased homogeneous radiopacity of normal-sized kidneys suggested the diagnosis of SO. Skeletal survey showed osteopenia and characteristic symmetrical metaphyseal transverse bands in long bones, progressively becoming more dense and migrating towards the diaphysis. Multiple pathological and slowly healing fractures of the limbs occurred at the dense band level. A radiopaque rim was then observed in flat bones, epiphyseal nuclei, and vertebral bodies. Inflammatory granulomatous reaction, induced by the presence of oxalate crystals in the marrow spaces, coexisted with progressively evident radiological signs of secondary hyperparathyroidism, with partially overlapping features. The patient was treated by peritoneal dialysis and hemodialysis until combined liver-kidney transplantation. There are no previous reports of infants treated with hemodialysis for more than 2 years. PMID- 19132373 TI - The microtubule-active antitumor compound TTI-237 has both paclitaxel-like and vincristine-like properties. AB - PURPOSE: To compare TTI-237 (5-chloro-6-[2,6-difluoro-4-[3 (methylamino)propoxy]phenyl]-N-[(1S)-2,2,2-trifluoro-1-methylethyl]-[1, 2, 4]triazolo[1,5-a]pyrimidin-7-amine butanedioate) with paclitaxel and vincristine in order to better understand the properties of this new anti-microtubule agent. METHODS: Tubulin polymerization and depolymerization were followed by turbidimetric assays. Effects of compounds on the binding of [(3)H]guanosine triphosphate ([(3)H]GTP) to tubulin were studied by competition binding assays. Effects of compounds on the phosphorylation of a panel of intracellular proteins were determined by flow cytometry using phosphoprotein-specific antibodies. RESULTS: At low molar ratios of TTI-237:tubulin heterodimer (about 1:30), TTI-237 enhanced depolymerization kinetics in response to low temperature, but stabilized the aggregates at higher ratios (about 1:4). Similarly, the aggregates induced in microtubule protein by TTI-237 were depolymerized by excess Ca(++) at low TTI 237:tubulin-heterodimer molar ratios, but were stable at higher ratios. TTI-237 inhibited the exchange of [(3)H]GTP at the exchangeable nucleotide site of the tubulin heterodimer, and was similar to vincristine in its effects on the phosphorylation of eight intracellular proteins in HeLa cells. CONCLUSIONS: TTI 237 has properties that distinguish it from typical vinca-site and taxoid-site ligands, and therefore it may exemplify a new class of microtubule-active compounds. PMID- 19132374 TI - Becatecarin (rebeccamycin analog, NSC 655649) is a transport substrate and induces expression of the ATP-binding cassette transporter, ABCG2, in lung carcinoma cells. AB - PURPOSE: ABCG2 overexpression has been linked to resistance to topoisomerase inhibitors, leading us to examine the potential interaction between ABCG2 and becatecarin. METHODS: Interaction with ABCG2 was determined by ATPase assay, competition of [(125)I]iodoarylazidoprazosin (IAAP) photolabeling and flow cytometry. Cellular resistance was measured in 4-day cytotoxicity assays. ABCG2 expression was measured by fluorescent-substrate transport assays and immunoblot. RESULTS: Becatecarin competed [(125)I]-IAAP labeling of ABCG2, stimulated ATPase activity and, at concentrations greater than 10 microM, inhibited ABCG2-mediated transport. Becatecarin-selected A549 Bec150 lung carcinoma cells were 3.1-, 15-, 8-, and 6.8-fold resistant to becatecarin, mitoxantrone, SN-38 and topotecan, respectively. A549 Bec150 cells transported the ABCG2 substrates pheophorbide a, mitoxantrone and BODIPY-prazosin and displayed increased staining with the anti ABCG2 antibody 5D3 compared to parental cells. Increased ABCG2 expression was confirmed by immunoblot. CONCLUSIONS: Our results suggest that becatecarin is transported by ABCG2 and can induce ABCG2 expression in cancer cells. PMID- 19132375 TI - [CUP syndrome: molecular pathogenesis and biology]. AB - Carcinoma of unknown primary (CUP) is an intriguing clinical finding defined as biopsy-proven metastasis from a malignancy in the absence of an identifiable primary site after a complete clinical workup. CUP is a relatively common clinical entity, accounting for approximately 3-5% of all cancer diagnoses, and consists of a heterogeneous group of tumors that have acquired the capacity to metastasize before the development of a clinically evident primary lesion. The mechanisms responsible for early metastasis and lack of a detectable primary tumor are largely unknown. Although remarkable tools have been developed for immunohistological classification of CUP on the basis of the likely tissue of origin, data on molecular pathogenesis and biology of this disorder are rare. A wide variety of chromosomal aberrations are seen in CUP, with aberrations of chromosomes 1, 6, 7, and 11 having been most frequently described. 66-75% of CUP express epidermal growth factor receptor while overexpression of Her2/neu seems to be rare. In contrast to most other tumor entities p53 mutations have been found only in a minority of CUP tumors. Recently, several independent studies have demonstrated proof of principle for the use of gene expression microarrays in identifying a primary site for CUP. Therefore, gene expression and also genomic profiling tools represent promising analytical approaches to assist with the management of CUP patients. PMID- 19132376 TI - Ectopic overexpression of AtHDG11 in tall fescue resulted in enhanced tolerance to drought and salt stress. AB - Tall fescue (Festuca arundinacea Schreb.) is a cool-season perennial grass, which has been conventionally grown in the temperate area. However, as a major type of cool-season turf grass, its growth has been extended to the sub-tropical climate or even to the transitional climate between the sub-tropical and the tropical, and, in some cases, to heavily salinized lands. The extended growth imposes a serious challenge to its tolerance to the abiotic stress, particularly to drought, salt and high temperature. Here, we report a successful introduction of Arabidopsis AtHDG11 into the tall fescue via Agrobacterium-mediated transformation. The ectopic overexpression of AtHDG11 under the control of CaMV 35S promoter with four enhancers resulted in significantly enhanced tolerance to drought and salt stress. No obvious adverse effects on growth and development were observed in the transgenic plants. The enhanced stress tolerance was associated with a more extensive root system, a lower level of malondialdehyde, a nearly normal Na(+)/K(+) ratio, a higher level of proline and a kinetically accelerated induction of SOD and CAT activities observed in the transgenic plants during drought and/or salt stress, indicating that an enhanced ROS scavenging capability might play a significant role in the acquired tolerance to the abiotic stress. PMID- 19132378 TI - Navigated femoral anteversion measurements: general precision and registration options. AB - INTRODUCTION: Intraoperative reduction of femoral fractures can result in rotational malalignment. Navigation modules allow fluoroscopy-based femoral anteversion (AV) measurements; however, their clinical feasibility has not been fully evaluated. An important technical consideration when obtaining navigated femoral rotational alignment is the necessity for orthogonal navigated fluoroscopic images. METHODS: The current investigators hypothesized that there would be a critical imaging angle between the fluoroscopic images which optimized the acquisition of accurate rotational measurements. Following initial testing in plastic femora, 14 intact human cadaveric femora were subjected to CT and navigated fluoroscopic assessment of AV. The navigated registration process included the following test series: Standard perpendicular AP and lateral imaging at a 90 degrees angle; planar rotation of the lateral fluoroscopy position perpendicular to the axis of femoral neck and angled to the AP position at 75 degrees, 60 degrees, and 50 degrees; inlet and outlet imaging of the axis of the femoral neck at angles of 10 degrees, 20 degrees, and 30 degrees. For all setups, the difference between the AV angle of the navigation system and the CT-measured angle was calculated. RESULTS: Results revealed no relevant differences between CT and navigated determination of plastic femora (1.5 degrees). No significant deviations of cadaver femora with 90 degrees angle imaging (mean 2.00 degrees, range 0.00 degrees-4.33 degrees), 75 degrees angle imaging (mean 1.95 degrees, range 0.00 degrees-3.33 degrees) and 60 degrees imaging (mean 2.00 degrees, range 0.00 degrees-3.33 degrees) were found. However, significant deviations were found for the navigated 50 degrees angled imaging technique (mean 5.02 degrees, range 2.33 degrees-7.67 degrees). CONCLUSION: These data demonstrated that the femoral AV angle can be calculated accurately by using two navigated fluoroscopic images when a minimum angle of 60 degrees is utilized between the angled fluoroscopic images for the registration process. A difference of <60 degrees does not allow for accurate navigated measurements. PMID- 19132377 TI - [Chemical and spectroscopic aspects of the application of dyes in vitreoretinal surgery]. AB - The advantages of the use of optical contrast agents in surgery, such as vitreoretinal microsurgery, are presented. Requirements concerning these materials and the potentials and limitations of their application are discussed on the basis of their coloristic properties. Applications of fluorescent dyes are examined. PMID- 19132379 TI - Complete septate uterus, obstructed hemivagina, and ipsilateral renal anomaly: pregnancy course complicated by a rare urogenital anomaly. AB - BACKGROUND: Abnormalities of the female reproductive tract are associated with a multitude of intrapartum complications. In the case that follows, we report the complicated pregnancy course of a woman diagnosed postpartum with the uncommon syndrome of obstructed hemivagina and ipsilateral renal anomaly (OHVIRA) associated with a complete septate uterus. CASE: A 19-year-old nulliparous woman with a history of a right nephrectomy secondary to multicystic dysplastic kidney disease presented with premature rupture of membranes at 30 weeks' gestation. After failed induction of labor at 34 weeks' gestation, a uterine anomaly was discovered at the time of cesarean section. Magnetic resonance imaging performed postpartum revealed a complete septate uterus, longitudinal vaginal septum, and obstructed right hemivagina. CONCLUSION: The OHVIRA syndrome is classically associated with uterus didelphys, not complete septate uterus as in this woman. This case illustrates the potential for intrapartum complications in women with abnormalities of the urogenital tract. PMID- 19132380 TI - A rare case of primary malignant melanoma of cervix. AB - Primary malignant melanoma (MM) of the uterine cervix is a rare neoplasm. Historically all MMs of the cervix were thought to be metastatic. But after the discovery of melanocytes in the cervix in 1959, it was recognized that primary MM of the cervix exists as a separate entity. The treatment of this condition is not yet standardized and the overall prognosis of these patients is very poor. We present a 40-year-old patient of primary MM of cervix including its diagnosis, management, follow-up and a brief review of literature. PMID- 19132381 TI - Fetal lung volume: comparison by 2D- and 3D-sonography in normal fetuses. AB - PURPOSE: To compare two-dimensional ultrasonography (2D US) and three-dimensional ultrasonography (3D US) in the assessment of normal fetal lung volume. METHODS: A cross-sectional study was performed involving 50 normal pregnancies at 24-32 weeks' gestation. The following equations were used for lung volume calculation by 2D US: Eq(2D1) = 4.24 + {1.53 x [(area of base of both lungs) x 1/3 (height of right lung)]} and Eq(2D2) = [anteroposterior diameter (X) x transverse diameter (Y) x cranial-caudal diameter (Z) of the right lung x 0.152 + (X') x (Y') x (Z') of the left lung x 0.167]. For 3D US, the virtual organ computer-aided analysis (VOCAL) method was used with a 30 degrees rotation angle and the total lung volume (V3D) was obtained by summing the volumes of each lung. Regression models (R (2)) were devised to assess lung volume evolution over the course of the pregnancy. Pearson's correlation coefficient (r) was used to assess correlation among the techniques, while Friedman's test was used for means comparisons. RESULTS: Strong correlation was observed among the three techniques [V3D vs. Eq(2D2) r = 0.856; V3D vs. Eq(2D1) r = 0.838 and Eq(2D2) vs. Eq(2D1) r = 0.964; all with P < 0.001]. Mean lung volumes were 37.05 +/- 9.67, 29.79 +/- 8.79 and 12.67 +/- 4.12 ml for V3D, Eq(2D1) and Eq(2D2), respectively (P < 0.001). CONCLUSIONS: Strong correlation and significant difference was observed among the three techniques of fetal lung volume assessment in normal fetuses. PMID- 19132382 TI - Thirteen X-chromosomal short tandem repeat loci multiplex data from Taiwanese. AB - Study results of variations in the X chromosome are useful tools in researching the genetic diversity of human populations and individual identification. We developed a 13 X chromosomal short tandem repeat (STR) multiplex system (DXS6807, DXS8378, DSX9902, DXS7132, DXS9898, DXS6809, DXS6789, DXS7424, DXS101, GATA172D05, HPRTB, DXS8377, DXS7423) amplified in one single polymerase chain reaction. DNA samples of 113 male and 108 female Taiwanese Han subjects were successfully analyzed using this 13 X-STR multiplex system. The distributions of allele frequencies were examined for independence. DXS8377, DXS101, DXS6789, and DXS6809 were found to be the most polymorphic markers in this study. High values of discrimination power and mean exclusion chance without significant evidence of association between these markers were obtained. In conclusion, this 13 X chromosomal STR multiplex system offers considerable forensic efficiency and may be useful in forensic identification casework. PMID- 19132383 TI - Long-term administration of tempol attenuates postinfarct ventricular dysfunction and sympathetic activity in rats. AB - The purpose of this study was to determine whether the long-term administration of tempol attenuates postinfarct ventricular dysfunction and sympathetic activity in rats. Myocardial infarction (MI) was induced by left descending coronary artery ligation. Tempol was orally administered in drinking water (2 mmol/L), which was initiated 4 h after infarction and continued for 6 weeks. Tempol prevented not only the increases in left ventricular end-diastolic pressure and volume but also the decreases in ejection fraction and peak velocities of contraction in MI rats. The treatment normalized the increased renal sympathetic nerve activity (RSNA) and plasma norepinephrine level, as well as the enhanced cardiac sympathetic afferent reflex (CSAR; an excitatory cardiovascular reflex partially contributing to the sympathetic activation in chronic heart failure) and the RSNA responses to microinjection of angiotensin II into paraventricular nucleus in MI rats. Furthermore, tempol prevented the increased AT(1) receptor protein expression and superoxide anion level in both paraventricular nucleus and rostral ventrolateral medulla in MI rats. In conclusion, long-term administration of tempol attenuates ventricular dysfunction and normalizes sympathetic neural control in MI rats. The normalization of the CSAR, levels of superoxide anions and AT(1) receptor expression, and the response to angiotensin II in the paraventricular nucleus and rostral ventrolateral medulla may partially contribute to the beneficial effects of tempol on central sympathetic control. PMID- 19132384 TI - Active neovascularization and possible vascular-centric development of gastric and periscapular elastofibromas. AB - An elastofibroma is a benign and rare fibrous lesion that most commonly occurs in the periscapular region. A gastrointestinal elastofibroma is extremely rare. In the present study, six cases of elastofibromas including a case in the stomach were evaluated. The gastric case revealed widely distributed lesions in the submucosal layer with perivascular fibrotic lesions (PVFLs) and some PVFLs were distributed to the skip lesions of elastofibroma. These PVFLs were also observed in all five periscapular cases and invariably contained elastic fibers which showed various degree of maturation. CD34-positive stromal cells were observed not only in elastofibromas but also in PVFLs in each case. These findings suggested the possibility of the PVFLs were the primary lesions of elastofibroma and their vascular-centric development. The percentage of the CD105-positive vessels in elastofibroma group was significantly higher than in the control group. This result indicates active neovascularization in elastofibromas. PMID- 19132385 TI - Neurotoxic lesions of the thalamic reuniens or mediodorsal nucleus in rats affect non-mnemonic aspects of watermaze learning. AB - Rats with bilateral neurotoxic reuniens (RE), mediodorsal (MD), hippocampal (HIPP) or sham (SH) lesions were tested in a standard watermaze task, together with unoperated rats. RE-rats and SH-controls readily learned to swim directly to a hidden platform. In contrast, MD-rats displayed a transient deficit characterized initially by thigmotaxis. Like in previous studies, HIPP-rats had long latencies throughout training and displayed more random swims than the other groups. In a memory probe test with the platform removed, SH- and RE-rats approached the correct location relatively directly but, whereas SH-controls persistently searched in the training quadrant, RE-rats switched to searching all over the pool. The MD-group swam in loops to the platform, but then displayed persistent searching in the training quadrant. The HIPP-group performed at chance. These distinct patterns indicate that, although their search strategies were different, RE- and MD-rats had acquired sufficient knowledge about the platform location and could recall information in the probe test. All groups performed well in a subsequent cue test with a visible platform, with RE-rats initially escaping faster than the SH- and HIPP-groups, and MD-rats improving from an initially poorer level of performance to control level. This indicates that there were no sensorimotor or motivational deficits associated with any of the lesions. In conclusion, while the RE and MD nuclei seem not to be critical for the learning and memory of a standard watermaze task, they may contribute to non-mnemonic strategy shifting when animals are challenged in ways that do not occur during training. PMID- 19132386 TI - Analysis of body posture in children with mild to moderate asthma. AB - The mechanical alterations related to the excessive use of accessory respiratory muscles and the mouth breathing observed in children with asthma may lead to the development of alterations in head posture, shoulders, thoracic region and, consequently, in alterations of body posture. The purpose of this study was to assess body posture changes of children with asthma compared to a non-asthmatic control group matched for gender, age, weight, and height. Thirty children with asthma and 30 non-asthmatic children aged 7 to 12 years were enrolled in this study. Digital photographic records were obtained for analysis of the body posture of the children by computed photogrammetry. The intraclass correlation coefficient and Student's t test (p < 0.05) were used for statistical analysis. There were no significant differences between groups for the angles analyzed, except for the knee flexor angle. These results demonstrate that children with asthma did not present postural alterations compared to non-asthmatic controls since the only angle for which there was a significant difference between groups showed weak reproducibility. The findings of this study do not support the notion that children with asthma present alterations in body posture. PMID- 19132387 TI - Ulceroglandular tularemia in a toddler in Germany after a mosquito bite. AB - INTRODUCTION: Although tularemia is a long-known disease, its significance had diminished over the last decades in Middle Europe. However, over the past years, there is new evidence suggesting that tularemia has re-emerged in Germany. In 2007, the highest number of human cases for almost 50 years has been notified. Beside typical vectors, new ways of transmission seem to gain significance. So far, mosquito bite-transmitted tularemia has only been known from Scandinavia but not from Middle Europe. CASE REPORT: We report the first case of a 1-year-old toddler from Southwestern Germany with mosquito bite-associated ulceroglandular tularaemia. The new and interesting features of this case are the young age of the patient and the unusual transmission route. The available data suggesting changes in the epidemiology for tularemia in Germany are reviewed. This is an interesting case of infantile tularemia with a very unusual transmission route, highlighting ongoing changes in the epidemiology of tularemia in Germany. PMID- 19132388 TI - Elevated PRL-3 expression was more frequently detected in the large primary gastric cancer and exhibits a poor prognostic impact on the patients. AB - PURPOSE: It has been reported that elevated PRL-3 expression was closely associated with lymph node metastasis of gastric cancer. In the present study, we detected the expression of PRL-3 in primary gastric cancer tissue, and evaluated its prognostic impact on the patients. METHODS: Total 137 gastric tumor samples were measured for PRL-3 phosphatase expression using immunohistochemistry method, and the overall survival rate was compared between the patients with high PRL-3 expression (n = 85) and those with moderate or low PRL-3 expression (n = 52) in the primary tumor. RESULTS: PRL-3 expression was more frequently detected in the tumors with a diameter >40 mm and in advanced stages (TNM stage III or IV). Furthermore, the overall survival rate of the patients with high expression of PRL-3 in the primary tumor was significantly less than those with moderate or low expression (P < 0.001). CONCLUSIONS: PRL-3 expression is associated with gastric cancer progression. Its high expression in the primary lesion had a negative impact on the prognosis of the patients. This strongly suggests that PRL-3 should be considered as a prognostic factor. PMID- 19132389 TI - Impact of pepsinogen C polymorphism on individual susceptibility to gastric cancer and its precancerous conditions in a Northeast Chinese population. AB - PURPOSE: Human pepsinogen C (PGC) is an aspartic protease produced specifically by the gastric mucosa, and is considered as a mature marker of gastric epithelium. This study examined the contributions of PGC polymorphisms and the Helicobacter pylori (H. pylori) infection to the risk of gastric cancer (GC), and its precancerous conditions in a Northeast Chinese population. METHODS: The PGC insertion/deletion polymorphism was evaluated by polymerase chain reaction analysis, followed by direct DNA sequencing in 564 cases of GC, atrophic gastritis (AG), gastric ulcer (GU) and superficial gastritis (as control). All cases were frequency-matched 1:1 by gender and age (+/-5). H. pylori infection was identified by serum anti-H. pylori IgG measurement through enzyme-linked immunosorbent assay. RESULTS: Patients with a homozygous PGC allele 1 genotype had a significant risk of AG [adjusted odds ratio (OR) 3.11; 95% confidence interval (CI) 1.44-6.71] or of GC (OR 3.00; 95% CI 1.38-6.51), and a significantly elevated risk of intestinal metaplasia (OR 1.90, 95% CI 1.11-3.27). PGC polymorphism with H. pylori infection increased risk of GU (OR 8.69; 95% CI 1.01-74.69), and AG (OR 11.12; 95% CI 1.37-90.84) or GC (OR 10.61; 95% CI 1.28 87.79) in a super-multiplicative manner. The S value was 5.40, 6.48 and 4.34; and the AP value was 72.09, 7.00 and 69.69%, respectively. CONCLUSIONS: The PGC gene polymorphism increases an individual's susceptibility to GC and its precancerous conditions. Moreover, the PGC gene polymorphism shows a positive link to H. pylori infection in the development of GC. PMID- 19132390 TI - Site influence in parasite distribution from fishes of the Bonifacio Strait Marine Reserve (Corsica Island, Mediterranean Sea). AB - The spatial distribution of parasite communities from teleost fishes (Diplodus vulgaris, Mullus surmuletus, Pagellus erythrinus, Phycis phycis, Scorpaena scrofa and Symphodus tinca) was studied in the Bonifacio Strait Marine Reserve (Corsica Island) with different protection levels. Canonical correspondence analysis was used to study spatial patterns and to quantify the importance of site influence on parasite communities. To supplement these results, we used the indicator species analysis (Indval) to identify parasite species as bioindicators of a given ecosystem. Moreover, fish spatial distribution in the marine reserve was evaluated in order to link parasite species occurrence and fish abundance. Several parasite species were associated with a specific site, and some of them were considered as site bioindicators, especially in the western part of the protected area (Bruzzi and Moines Islands). However, the relative abundance of fish followed a different spatial pattern. Stronger fish abundance was observed in the eastern part of the reserve (Lavezzi Islands). The parasite species were correlated to sites inside as well outside the reserve. The parasite distribution seemed not to be influenced by the protection status of the site. Furthermore, in our study, most of the parasite species were not linked to a site. The most important factor influencing their distribution was host specificity. This suggested that the parasite life traits history remained a fundamental determinant in the community composition. PMID- 19132393 TI - Faithful inheritance of cytosine methylation patterns in repeated sequences of the allotetraploid tobacco correlates with the expression of DNA methyltransferase gene families from both parental genomes. AB - The widespread occurrence of epigenetic alterations in allopolyploid species deserves scrutiny that DNA methylation systems may be perturbed by interspecies hybridization and polyploidization. Here we studied the genes involved in DNA methylation in Nicotiana tabacum (tobacco) allotetraploid containing S and T genomes inherited from Nicotiana sylvestris and Nicotiana tomentosiformis progenitors. To determine the inheritance of DNA methyltransferase genes and their expression patterns we examined three major DNA methyltransferase families (MET1, CMT3 and DRM) from tobacco and the progenitor species. Using Southern blot hybridization and PCR-based methods (genomic CAPS), we found that the parental loci of these gene families are retained in tobacco. Homoeologous expression was found in all tissues examined (leaf, root, flower) suggesting that DNA methyltransferase genes were probably not themselves targets of uniparental epigenetic silencing for over thousands of generations of allotetraploid evolution. The level of CG and CHG methylation of selected high-copy repeated sequences was similar and high in tobacco and its diploid progenitors. We speculate that natural selection might favor additive expression of parental DNA methyltransferase genes maintaining high levels of DNA methylation in tobacco, which has a repeat-rich heterochromatic genome. PMID- 19132394 TI - The application of expression analysis in elucidating the eukaryotic elongation factor one alpha gene family in Arabidopsis thaliana. AB - Eukaryotic elongation factor one alpha (eEF1A) encoding genes are part of the large GTP binding protein family. The eEF1A family is important for protein synthesis and actin filament and bundle formation. In this study, the expression of four eEF1A genes in Arabidopsis thaliana is reported. Microarray analyses of the gene family showed high expression levels in germinating seeds, embryos, and shoot and root meristems. Quantitative real time RT-PCR was used to determine individual eEF1A gene expression. Unlike animals, in Arabidopsis tissues all four eEF1A genes were expressed in all tissues sampled. However, the abundance of each transcript varied spatially. Knocking out expression of one eEF1A gene produced seedlings with stunted roots and a subsequent change in expression of the other three eEF1A genes. The varying abundance of each gene in different tissues may indicate different concentration requirements for each message product. These results will be very useful for elucidating the role of each gene in growth, development, and stress responses of the plant. PMID- 19132395 TI - Uterine leiomyomata and decreased height: a common HMGA2 predisposition allele. AB - Uterine leiomyomata (UL) are the most common female pelvic tumors and the primary indication for hysterectomy in the United States. We assessed genetic liability for UL by a known embryonic proliferation modulator, HMGA2, in 248 families ascertained through medical record-confirmed affected sister-pairs. Using a (TC)( n ) repeat in the 5' UTR and 17 SNPs spanning HMGA2, permutation-based association tests identified a significant increase in transmission of a single TC repeat allele (TC227) with UL (allele-specific P = 0.00005, multiple testing corrected min-P = 0.0049). The hypothesis that TC227 is a pathogenic variant is supported by a trend towards higher HMGA2 expression in TC227 allele-positive compared with non-TC227 UL tissue as well as by absence of culpable exonic sequence variants. HMGA2 has also been suggested recently by three genome-wide SNP studies to influence human height variation, and our examination of the affected sister-pair families revealed a significant association of TC227 with decreased height (allele-specific P = 0.00033, multiple testing corrected min-P = 0.016). Diminished stature and elevated risk of UL development have both been correlated with an earlier age of menarche, which may be the biological mechanism for TC227 effects as a tendency of women with TC227 to have an earlier onset of menarche was identified in our study population. These results indicate HMGA2 has a role in two growth-related phenotypes, UL predisposition and height, of which the former may affect future medical management decisions for many women. PMID- 19132396 TI - Venom apparatus of the Brazilian tarantula Vitalius dubius Mello-Leitao 1923 (Theraphosidae). AB - Tarantula venoms are a cocktail of proteins and peptides that have been increasingly studied in recent years. In contrast, less attention has been given to analyzing the structure of the paired cephalic glands that produce the venom. We have used light, electron, and confocal microscopy to study the organization and structure of the venom gland of the Brazilian tarantula Vitalius dubius. The chelicerae are hairy chitinous structures, each with a single curved hollow fang that opens via an orifice on the anterior surface. Internally, each chelicera contains striated muscle fiber bundles that control fang extension and retraction, and a cylindrical conical venom gland surrounded by a thick well developed layer of obliquely arranged muscle fibers. Light microscopy of longitudinal and transverse sections showed that the gland secretory epithelium consists of a sponge-like network of slender epithelial cell processes with numerous bridges and interconnections that form lacunae containing secretion. This secretory epithelium is supported by a basement membrane containing elastic fibers. The entire epithelial structure of the venom-secreting cells is reinforced by a dense network of F-actin intermediate filaments, as shown by staining with phalloidin. Neural elements (axons and acetylcholinesterase activity) are also associated with the venom gland. Transmission electron microscopy of the epithelium revealed an ultrastructure typical of secretory cells, including abundant rough and smooth endoplasmic reticulum, an extensive Golgi apparatus, and numerous mitochondria. PMID- 19132397 TI - Starvation induces apoptosis in the midgut nidi of Periplaneta americana: a histochemical and ultrastructural study. AB - The effects of starvation on cell death in the midgut of Periplaneta americana were studied histochemically and ultrastructurally. TUNEL assays showed that cell death began to increase in the columnar cells and nidi, the nests of stem cells and newborn cells from 2 weeks of starvation. A significant increase in cell death occurred in the nidi after 4 weeks of starvation. Cockroaches starved for 4 weeks showed active-caspase-3-like immuno-reactivity both in the columnar cells and nidi, whereas control cockroaches that were fed for 4 weeks showed this reactivity only in the apical cytoplasm of columnar cells. Electron microscopy revealed no chromatin condensation in the nucleus of columnar cells of cockroaches, whether fed or starved for 4 weeks. Starved cockroaches exhibited many small vacuoles in the cytoplasm of some columnar cells and "floating" organelles including nuclei in the lumen. A 4-week starvation induced the appearance of cytoplasmic fragmentation and secondary lysosomes in the nidi. Each fragment contained nuclear derivatives with condensed chromatin, i.e. apoptotic bodies. Mitotic cells were found in some, but not all nidi, even within the same starved sample. Fragmentation was not observed in the nidi of control cockroaches. Thus, starvation increases cell death not only in the columnar cells, but also in the nidi. The cell death in the nidi is presumably apoptosis executed by caspase 3. PMID- 19132398 TI - Phosphorus reserves increase grass regrowth after defoliation. AB - Accumulation of P above levels that promote growth, a common plant response called "luxury consumption", can be considered as a form of reserve to support future growth when the nutrient can subsequently be mobilized. However, the effect of P reserves on regrowth following defoliation has not been demonstrated. We tested the hypothesis that P luxury consumption increases plant tolerance to defoliation. We performed two experiments with four grass species from a continuously grazed temperate grassland in the Flooding Pampa (Argentina). The first experiment, aimed at generating P luxury consumption by fertilization, resulted in one species (Sporobolus indicus) showing luxury consumption. In this way, we were able to obtain plants of S. indicus with similar biomass but contrasting amounts of P reserves. The second experiment evaluated the subsequent regrowth following defoliation on a P-free medium of these plants differing in P reserves. Regrowth was larger for plants that had shown P luxury consumption during a previous period than for plants with lower levels of P reserves. During regrowth these plants showed a clear pattern of P remobilization from the stubble, crown, and root compartments to the regrowing tissue, in addition to a likely reutilization of P present in leaf-growth zones. This work is the first showing that high levels of P reserves can confer tolerance to defoliation by promoting compensatory growth under P deficiency. PMID- 19132399 TI - Evidence of rapid shifts in the trophic base of lotic predators using experimental dietary manipulations and assimilation-based analyses. AB - Assimilation-based techniques such as stoichiometric analysis, bulk tissue stable isotope analysis (SIA), fatty acid (FA) profiling and compound-specific SIA of FAs can be used to resolve ambiguities in consumer-resource relationships, but comparisons of their effectiveness are lacking. Feeding trials and concurrent application of these techniques were used to investigate the trophic base of two shrimps from Hong Kong streams: the atyid Caridina cantonensis, which is a primary consumer, and predatory Macrobrachium hainanense (Palaemonidae). Leaf litter and periphyton were fed to C. cantonensis (CC-LF and CC-PF, respectively) reared in the laboratory for 2 months, when C. cantonensis, that had fed on a mixed diet (CC-WC), were collected from the field. Atyids from each group (CC-LF, CC-PF and CC-WC) were fed to M. hainanense (MH-L, MH-P and MH-W, respectively) during a further 2-month trial, at the end of which M. hainanense were also collected from the field (MH-R). FA biomarkers present in CC-WC and CC-PF indicated that C. cantonensis depended primarily on autochthonous foods, and FA profiles of CC-WC were distinct from those of CC-LF. Differences in C/N ratios and isotope signatures of leaf litter and periphyton were not reflected in tissues of atyids or palaemonid predators. FA profiles of M. hainanense groups were similar, but FA stable C isotope ratio (delta(13)C) signatures of MH-R and MH-W were distinct from those of MH-L and leaf litter. FA delta(13)C signatures of MH-R were similar to those of MH-P and MH-W, indicating that autochthonous resources constituted the trophic base of production for this predator. This is the first study using compound-specific SIA to study stream food chains. Compound specific SIA and FA profiling allowed accurate elucidation of consumer-resource relationships that were not revealed by stoichiometry or bulk tissue SIA. Compound-specific SIA was particularly sensitive for detecting rapid shifts in the predator trophic base. This tool will have wide applicability for investigating food webs in a range of ecosystems. PMID- 19132400 TI - Frequent fire promotes diversity and cover of biological soil crusts in a derived temperate grassland. AB - The intermediate disturbance hypothesis (IDH) predicts that species diversity is maximized at moderate disturbance levels. This model is often applied to grassy ecosystems, where disturbance can be important for maintaining vascular plant composition and diversity. However, effects of disturbance type and frequency on cover and diversity of non-vascular plants comprising biological soil crusts are poorly known, despite their potentially important role in ecosystem function. We established replicated disturbance regimes of different type (fire vs. mowing) and frequency (2, 4, 8 yearly and unburnt) in a high-quality, representative Themeda australis-Poa sieberiana derived grassland in south-eastern Australia. Effects on soil crust bryophytes and lichens (hereafter cryptogams) were measured after 12 years. Consistent with expectations under IDH, cryptogam richness and abundance declined under no disturbance, likely due to competitive exclusion by vascular plants as well as high soil turnover by soil invertebrates beneath thick grass. Disturbance type was also significant, with burning enhancing richness and abundance more than mowing. Contrary to expectations, however, cryptogam richness increased most dramatically under our most frequent and recent (2 year) burning regime, even when changes in abundance were accounted for by rarefaction analysis. Thus, from the perspective of cryptogams, 2-year burning was not an adequately severe disturbance regime to reduce diversity, highlighting the difficulty associated with expression of disturbance gradients in the application of IDH. Indeed, significant correlations with grassland structure suggest that cryptogam abundance and diversity in this relatively mesic (600 mm annual rainfall) grassland is maximised by frequent fires that reduce vegetation and litter cover, providing light, open areas and stable soil surfaces for colonisation. This contrasts with detrimental effects of 2-year burning on native perennial grasses, indicating that this proliferation of cryptogams has potentially high functional significance for situations where vegetation cover is depleted, particularly for reducing soil erosion. PMID- 19132401 TI - Integrating edge effects into studies of habitat fragmentation: a test using meiofauna in seagrass. AB - Habitat fragmentation is thought to be an important process structuring landscapes in marine and estuarine environments, but effects on fauna are poorly understood, in part because of a focus on patchiness rather than fragmentation. Furthermore, despite concomitant increases in perimeter:area ratios with fragmentation, we have little understanding of how fauna change from patch edges to interiors during fragmentation. Densities of meiofauna were measured at different distances across the edges of four artificial seagrass treatments [continuous, fragmented, procedural control (to control for disturbance by fragmenting then restoring experimental plots), and patchy] 1 day, 1 week and 1 month after fragmentation. Experimental plots were established 1 week prior to fragmentation/disturbance. Samples were numerically dominated by harpacticoid copepods, densities of which were greater at the edge than 0.5 m into patches for continuous, procedural control and patchy treatments; densities were similar between the edge and 0.5 m in fragmented patches. For taxa that demonstrated edge effects, densities exhibited log-linear declines to 0.5 m into a patch with no differences observed between 0.5 m and 1 m into continuous treatments. In patchy treatments densities were similar at the internal and external edges for many taxa. The strong positive edge effect (higher densities at edge than interior) for taxa such as harpacticoid copepods implies some benefit of patchy landscapes. But the lack of edge effects during patch fragmentation itself demonstrates the importance of the mechanisms by which habitats become patchy. PMID- 19132402 TI - Environmental factors influencing adult sex ratio in Trinidadian guppies. AB - Sex ratios can influence mating behaviour, population dynamics and evolutionary trajectories; yet the causes of natural sex ratio variation are often uncertain. Although secondary (birth) sex ratios in guppies (Poecilia reticulata) are typically 1:1, we recorded female-biased tertiary (adult) sex ratios in about half of our 48 samples and male-biased sex ratios in none of them. This pattern implies that some populations experience male-biased mortality, perhaps owing to variation in predation or resource limitation. We assessed the effects of predation and/or inter-specific resource competition (intraguild predation) by measuring the local catch-per-unit-effort (CPUE) of species (Rivulus killifish and Macrobrachium prawns) that may differentially prey on male guppies. We assessed the effects of resource levels by measuring canopy openness and algal biomass (chlorophyll a concentration). We found that guppy sex ratios were increasingly female-biased with increasing CPUE of Macrobrachium, and perhaps also Rivulus, and with decreasing canopy openness. We also found an interaction between predators and resource levels in that the effect of canopy openness was greatest when Macrobrachium CPUE was highest. Our study thus also reveals the value of simultaneously testing multiple environmental factors that may drive tertiary sex ratio variation. PMID- 19132403 TI - Plant-hummingbird interactions in the West Indies: floral specialisation gradients associated with environment and hummingbird size. AB - Floral phenotype and pollination system of a plant may be influenced by the abiotic environment and the local pollinator assemblage. This was investigated in seven plant-hummingbird assemblages on the West Indian islands of Grenada, Dominica and Puerto Rico. We report all hummingbird and insect pollinators of 49 hummingbird-pollinated plant species, as well as six quantitative and semi quantitative floral characters that determine visitor restriction, attraction and reward. Using nonmetric multidimensional scaling analysis, we show that hummingbird-pollinated plants in the West Indies separate in floral phenotypic space into two gradients-one associated with the abiotic environment and another with hummingbird size. Plants pollinated by large, long-billed hummingbirds had flowers with long corolla tube, large amounts of nectar and showy orange-red colouration. These attracted few or no insect species, whereas plants pollinated by small, short-billed hummingbirds were frequently pollinated by insects, particularly lepidopterans. The separation of plants related to environmental factors showed that species in the wet and cold highlands produced large amounts of dilute nectar, possessed no or a weak odour, and were associated with few insects, particularly few hymenopterans, compared to plants in the dry and warm lowlands. The most specialised hummingbird-pollinated plants are found in the West Indian highlands where they are pollinated by mainly large, long-billed hummingbirds. At the other extreme, highly generalised plants growing in the dry and warm lowlands are pollinated by small, short-billed hummingbirds and numerous insect species. This illustrates that, even within the hummingbird-pollinated flora, pollination syndrome and the degree of specialisation may vary tremendously depending on pollinator morphology and environment. PMID- 19132404 TI - Differential impacts of plant interactions on herbaceous species recruitment: disentangling factors controlling emergence, survival and growth of seedlings. AB - Recruitment is a crucial event in the plant life cycle that is very sensitive to interaction with established vegetation. Based on a large comparative experiment, we tested the hypothesis that the components of recruitment--emergence time and rate, seedling survival and biomass--differ in response to plant-plant interactions during recruitment. The consequences for the population are predicted with a simple demographic model assessing the response of seed production. In a common garden experiment, we recorded the recruitment of four target species in an individual-based survey protocol. A total of 7,680 seeds were sown within 20 neighbourhoods, consisting of 19 mono-specific herbaceous stands and a control treatment without vegetation. We measured transmitted light, temperature and moisture at soil surface to characterise the environmental conditions within neighbourhoods. The mean height of neighbours controlled temperature buffering and light interception and thus depicted the interaction gradient. Emergence rate and time increased with neighbour height in two of the four target species, while seedling survival and biomass significantly decreased with neighbour height in three and all four target species, respectively. We recorded a shift in seedling neighbour interactions under the tallest neighbours that largely favoured emergence but strongly depressed seedling survival and biomass. The components of recruitment were predicted to differ in their impact on later adult performance. Biomass strongly contributed to predicted seed production in three target species, and emergence had an equal or greater impact on a fourth species. These results confirm the fundamental role of plant-plant interactions in the recruitment of herbaceous species through a complex combination of habitat amelioration, which facilitates emergence and light competition, which in turn limits seedling survival and biomass. PMID- 19132405 TI - Rearing environment effects on immune defence in blue tit Cyanistes caeruleus nestlings. AB - Rearing conditions may influence ontogeny and functioning of the immune system. Activation of different mechanisms involved in host disease resistance and their internal regulation can be affected by intrinsic and extrinsic factors influencing development. I investigated how rearing environment can influence associations between humoral and cellular constituents of immune defence in nestling blue tits (Cyanistes caeruleus). The ability to mount a cell-mediated immune response was estimated as a hypersensitivity reaction to phytohaemagglutinin, and the ontogeny of humoral immunity was determined by assessing circulating levels of total IgG in 15-day-old nestlings. Heterogeneity in rearing conditions was evoked by placing nest-boxes in areas differing in habitat structural characteristics, and examining natural variation in nest ectoparasite infestations, hatching date, brood size and brood sex-ratio. Habitat characteristics, parasitism and hatching date may shape associations between different components of the immune system in developing birds. I discuss the effects of rearing conditions on the interaction between different arms of the immune system and the implications for understanding negative correlations within the immune system at the individual and brood level. PMID- 19132406 TI - Nonlinear continuum of egg size-number trade-offs in a snake: is egg-size variation fitness related? AB - The relationship between offspring size and offspring number is crucial to life history evolution. To examine how these two life history variables are coupled and whether an altered balance between them will result in changes in maternal fitness, we manipulated clutch size of the Chinese cobra (Naja atra) by using the techniques of hormonal manipulation and follicle ablation. Females receiving exogenous follicle-stimulating hormone produced more but smaller eggs, and females undergoing follicle ablation produced fewer but larger eggs. Neither body size (body mass and snout-vent length) at hatching nor egg mass at oviposition had a role in determining hatchling survival and growth. Female hatchlings were more likely to die in early post-hatching days and grew more slowly than male hatchlings. Our data show that: (1) there is a nonlinear continuum of egg size number trade-offs in N. atra within which there is a single inflexion where the rate at which egg size decreases with increasing clutch size, or clutch size increases with decreasing egg size, is maximized; (2) there is a fixed upper limit to egg size for a given-sized female, and the limit is not determined by her body volume; (3) egg size has no role in determining hatchling survival and growth; and (4) the extent to which females may enjoy reproductive benefits in a given reproductive episode depends on how well egg size and egg number are balanced. PMID- 19132407 TI - Morphological and physiological variation among seagrass (Zostera marina) genotypes. AB - Intraspecific variation in habitat-forming species can have important ecological consequences at the population, community, and ecosystem level. However, the contribution of genetic variation among individuals to these effects is seldom documented. We quantified morphological and physiological variation among genotypes of a marine foundation species, the seagrass Zostera marina. We grew replicate shoots of eight genetically distinct Zostera individuals collected from Bodega Bay, California, in a common garden environment and then quantified shoot production and morphology, nutrient uptake, and key photosynthetic parameters. We found that genotypes differed in shoot production, biomass, and both root and shoot nutrient uptake rates, even when corrected for genotype-specific biomass differences. In addition, the rank order of uptake ability differed for ammonium and nitrate, indicating that genotypes may exhibit resource partitioning of different forms of nutrients. Our results suggest that both niche complementarity among genotypes and the sampling/selection effect could contribute to previously observed positive effects of seagrass clonal diversity on resource utilization and biomass production. Further, they highlight that genotypic variation in key traits of habitat-forming species could have measurable effects on community structure and function. PMID- 19132408 TI - Pacific salmon effects on stream ecosystems: a quantitative synthesis. AB - Pacific salmon (Oncorhynchus spp.) disturb sediments and fertilize streams with marine-derived nutrients during their annual spawning runs, leading researchers to classify these fish as ecosystem engineers and providers of resource subsidies. While these processes strongly influence the structure and function of salmon streams, the magnitude of salmon influence varies widely across studies. Here, we use meta-analysis to evaluate potential sources of variability among studies in stream ecosystem responses to salmon. Results obtained from 37 publications that collectively included 79 streams revealed positive, but highly inconsistent, overall effects of salmon on dissolved nutrients, sediment biofilm, macroinvertebrates, resident fish, and isotopic enrichment. Variation in these response variables was commonly influenced by salmon biomass, stream discharge, sediment size, and whether studies used artificial carcass treatments or observed a natural spawning run. Dissolved nutrients were positively related to salmon biomass per unit discharge, and the slope of the relationship for natural runs was five to ten times higher than for carcass additions. Mean effects on ammonium and phosphorus were also greater for natural runs than carcass additions, an effect attributable to excretion by live salmon. In contrast, we observed larger positive effects on benthic macroinvertebrates for carcass additions than for natural runs, likely because disturbance by live salmon was absent. Furthermore, benthic macroinvertebrates and biofilm associated with small sediments (<32 mm) displayed a negative response to salmon while those associated with large sediments (>32 mm) showed a positive response. This comprehensive analysis is the first to quantitatively identify environmental and methodological variables that influence the observed effects of salmon. Identifying sources of variation in salmon-stream interactions is a critical step toward understanding why engineering and subsidy effects vary so dramatically over space and time, and toward developing management strategies that will preserve the ecological integrity of salmon streams. PMID- 19132409 TI - Investigating the adaptive model of thermal comfort for naturally ventilated school buildings in Taiwan. AB - Divergence in the acceptability to people in different regions of naturally ventilated thermal environments raises a concern over the extent to which the ASHRAE Standard 55 may be applied as a universal criterion of thermal comfort. In this study, the ASHRAE 55 adaptive model of thermal comfort was investigated for its applicability to a hot and humid climate through a long-term field survey performed in central Taiwan among local students attending 14 elementary and high schools during September to January. Adaptive behaviors, thermal neutrality, and thermal comfort zones are explored. A probit analysis of thermal acceptability responses from students was performed in place of the conventional linear regression of thermal sensation votes against operative temperature to investigate the limits of comfort zones for 90% and 80% acceptability; the corresponding comfort zones were found to occur at 20.1-28.4 degrees C and 17.6 30.0 degrees C, respectively. In comparison with the yearly comfort zones recommended by the adaptive model for naturally ventilated spaces in the ASHRAE Standard 55, those observed in this study differ in the lower limit for 80% acceptability, with the observed level being 1.7 degrees C lower than the ASHRAE recommended value. These findings can be generalized to the population of school children, thus providing information that can supplement ASHRAE Standard 55 in evaluating the thermal performance of naturally ventilated school buildings, particularly in hot-humid areas such as Taiwan. PMID- 19132410 TI - Incorporating diffuse photosynthetically active radiation in a single-leaf model of canopy photosynthesis for a 56-year-old Douglas-fir forest. AB - A simple top-down model of canopy photosynthesis (P) was developed and tested in this study. The model (referred to as the Q(e)-MM model) is P = alphaQ (e) P (max)/(alphaQ ( e ) + P (max)), alpha and P (max) are quantum-use efficiency and potential P, respectively. Q (e) is given by Q (d) (0) + kQ (b) (0), where Q (d) (0) and Q (b) (0) are the diffuse and direct photosynthetically active radiation (PAR) incident on the canopy, respectively. Q (e) can be considered to be the effective incident PAR contributing to P and k is a measure of the contribution of Q (b) (0) to Q (e). When k = 1, the Q(e)-MM model becomes the regular Michaelis-Menten type model of P (referred to as the MM model). A major objective of this study was to determine how well the Q(e)-MM model could estimate P of a 56-year-old coastal Douglas-fir stand. To this end, we parameterized the Q(e)-MM model using five and half years of eddy-covariance measurements of CO(2) flux above the Douglas-fir stand. The Q(e)-MM model, with the incorporation of a function of air temperature, accounted for 74% of the variance in over 34,000 half-hourly P measurements. P estimated using the Q(e)-MM model had no systematic errors with respect to Q (d) (0). Although the Q(e)-MM model has only one more parameter than the MM model, it accounted for 30% more variance in P than the latter when total incident PAR exceeded 900 micromol m(-2) s(-1). On average, k was found to be 0.22. We show that this small value of k reflects the significant effect of the scattering of the solar beam and the fraction of light-limited sunlit leaves. We also show that the success of the Q(e)-MM model was due to the fact that a large fraction of the sunlit leaves were light-limited as a result of their orientation to the solar beam. PMID- 19132411 TI - "When will I recover?" A national survey on patients' and physicians' expectations concerning the recovery time for acute back pain. AB - The objective of this study was to compare patients' and physicians' expectations concerning the recovery time for acute back pain, and their determinants. A French national observational survey was performed between October and December 2005. Each physician was asked to include the first three patients aged between 20 and 70 years presenting with acute back pain (VAS > 40 mm) of less than 1 month's duration. A total number of 1982 patients, with a mean age of 48.4 +/- 11.8 years (52.2% men), were enrolled by 834 physicians. Patients and their physicians expected recovery to take the same amount of time in 60.3% of cases (Kappa = 0.43). In 17.4% of cases, patients predicted a shorter recovery time than physicians, and in 22.4% of cases, patients predicted a longer recovery time. Both patients and physicians expected recovery time to be longer in women, and in older, unmarried, obese, and non working patients. Expected recovery time was also longer in patients with no sports activities, low back pain (as opposed to pain at other sites), a high level of disability, a history of back pain, and no triggering factor. The intensity of pain and the number of days of sick leave taken did not affect the patient's prediction of recovery time, but did affect the physicians' expectations. Physicians considered professional status, analgesic intake and previous sick leave to be independent predictive factors, whereas patients did not. Expected recovery time, assessed shortly after the onset of acute back pain is influenced by most of the clinical and demographic factors usually considered risk factors for chronic low back pain. Patients' predictions of recovery time should be assessed in acute back pain, to identify risks for expected chronicity. PMID- 19132413 TI - Single-level cervical radiculopathy: clinical outcome and cost-effectiveness of four techniques of anterior cervical discectomy and fusion and disc arthroplasty. AB - Although there are several accepted methods of surgical treatment for single level cervical radiculopathy, the choice depend on the surgeon's preference. The techniques may vary in peri-operative morbidity, short- and long-term outcome, but no study so far has analyzed their cost-effectiveness. This study might give some insight in balancing cost and effectiveness and deciding the right technique. Sixty consecutive patients (15 each group), mean age 36 (range 24-76 years) with single-level cervical disc disease underwent surgical treatment with four different techniques in two centers over the period of 1999-2005. The four groups were--(1) plate and tricortical autograft, (2) plate, cage, and bone substitute, (3) cage only, and (4) disc arthroplasty. The data was collected prospectively according to our protocol and subsequently analyzed. The clinical outcome was assessed comparing visual analog scale (VAS) of neck pain and, short form 12 (SF12) questionnaire both pre- and postoperatively. The radiological assessment was done for fusion rate and postoperative related possible complications at 3 months, 6 months, 1 year, and final follow-up. The cost analysis was done calculating the operative time, hospital stay, implant cost together. The mean follow-up period was 31 months (range 28-43 months). The clinical outcome in terms of VAS of neck and arm pain and SF12 physical and mental score improvement (P=0.001) were comparable with all four techniques. The radiological fusion rate was comparable to current available data. As the hospital stay was longer (average 5 days) with plate and autograft group, the total cost was maximum (average 2,920 pound sterling) with this group. There was satisfactory clinical and radiological outcome with all four techniques. Using the cage alone was the most cost-effective technique, but the disc arthroplasty was comparable to the use of cage and plate. Anterior cervical discectomy and fusion is an established surgical treatment for cervical radiculopathy. Single level cervical radiculopathy was treated with four different techniques. The clinical outcome and cost-effectiveness were compared in this study. PMID- 19132412 TI - Timing of surgery for sciatica: subgroup analysis alongside a randomized trial. AB - Surgery speeds up recovery for sciatica. Prolonged conservative care with surgery for those patients with persistent sciatica however, yields similar results at 1 year. To investigate whether baseline variables modify the difference in recovery rates between these treatment strategies, baseline data of 283 patients enrolled in a randomized trial, comparing early surgery with prolonged conservative care, were used to analyse effect modification of the allotted treatment strategy. For predictors shown to modify the effect of the treatment strategy, repeated measurement analyses with the Roland Disability Questionnaire and visual analogue scale pain as continuous outcomes were performed for every level of that predictor. Presumed predictive variables did not have any interaction with treatment, while "sciatica provoked by sitting" showed to be a significant effect modifier (P = 0.07). In a Cox model we estimated a hazard ratio (HR, surgery versus conservative) of 2.2 (95% CI 1.7-3.0) in favour of surgery when sciatica was provoked by sitting, while the HR was 1.3 (95% CI 0.8-2.2) when this sign was absent. The interaction effect is marginally significant (interactions are usually tested at the 10% level) but the patterns generated by the repeated measurement analyses of all primary outcomes are completely consistent with the inferred pattern from the survival analysis. Classical signs did not show any contribution as decision support tools in deciding when to operate for sciatica, whereas treatment effects of early surgery are emphasized when sciatica is provoked by sitting and negligible when this symptom is absent. PMID- 19132414 TI - Morphology of the smear layer after the application of simplified self-etch adhesives on enamel and dentin surfaces created with different preparation methods. AB - Mild self-etching adhesive systems modify and/or incorporate the smear layer into the resin-infiltrated demineralised dentin. Some factors such as type of bur and use of water spray might affect the thickness of the smear layer on substrates, enamel and dentin. Because of this, the present study evaluated the thickness of smear layers created by different finishing procedures, after the application of three simplified self-etching primers (Adper Prompt L-Pop and two experimental formulations) on enamel and dentin. After the application and removal of the primers' resinous component, the specimens were prepared for examination under a scanning electron microscope. Smear layers were thicker on enamel than on dentin, irrespective of the finishing methods used. Therefore, different thicknesses of smear layer on enamel/dentin might be an important factor to consider when evaluating the bonding efficacy of self-etching adhesives to both tooth substrates. PMID- 19132415 TI - Solvent impact on the aromaticity of benzene analogues: implicit versus explicit solvent approach. AB - Solvent impact on the structural index of aromaticity was modelled by polarised continuum field approximation (IEFPCM) and hybrid quantum chemistry (QM/MM) method. Significant solvent related relaxation of the solutes geometries were noticed especially for highly polar species. The significant reduction of the aromaticity was observed for some aromatic compounds in water solution compared to gas phase. The rationale of this fact was provided based on dipole moment changes, energy penalty for polarisation of solute and the distribution of frontier orbital densities. The incoherent predictions of explicit and implicit solvation models are noticed since in some cases the PCM approach artificially exaggerate the geometry relaxation in solution which is not observed if explicit solvent molecules are taken into account. PMID- 19132416 TI - CoMFA and CoMSIA analysis of 2,4-thiazolidinediones derivatives as aldose reductase inhibitors. AB - Diabetes remains a life-threatening disease. The clinical profile of diabetic subjects is often worsened by the presence of several long-term complications, for example neuropathy, nephropathy, retinopathy, and cataract. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on a series of 2,4-thiazolidinediones derivatives as aldose reductase (ALR2) inhibitors. Molecular ligand superimposition on a template structure was finished by the database alignment method. The 3D-QSAR models resulted from 44 molecules gave q (2) values of 0.773 and 0.817, r (2) values of 0.981 and 0.979 for CoMFA and CoMSIA, respectively. The contour maps from the models indicated that a large volume group next to the R-substituent will increase the ALR2 inhibitory activity. In fact, adding a CH(2)COOH substituent at the R-position would generate a new compound with higher predicted activity. PMID- 19132417 TI - Structural and energetic heterogeneities of canonical and oxidized central guanine triad of B-DNA telomeric fragments. AB - The intermolecular interaction energies in central guanine triad of telomeric B DNA were estimated based on ab initio quantum chemistry calculations on the MP2/aDZ level of theory. The source of structural information was molecular dynamics simulation of both canonical (AGGGTT) and oxidized (AG8oxoGGTT) telomere units. Our calculations demonstrate that significant stiffness of central triad occurs if 8oxoG is present. The origin of such feature is mainly due to the increase of stacking interactions of 8oxoG with neighbouring guanine molecules and stronger hydrogen bonding formation of 8oxoG with cytosine if compared with canonical guanine. Another interesting observation is the context independence of stacking interactions of 8oxoG. Unlike to 5'-G2/G3-3' and 5'-G3/G4-3' sequences which are energetically different, 5'-G2/8oxoG3-3' and 5'-8oxoG3/G4-3' sequences are almost iso-energetic. PMID- 19132419 TI - A systematic comparison of all mutations in hereditary sensory neuropathy type I (HSAN I) reveals that the G387A mutation is not disease associated. AB - Hereditary sensory neuropathy type 1 (HSAN I) is an autosomal dominant inherited neurodegenerative disorder of the peripheral nervous system associated with mutations in the SPTLC1 subunit of the serine palmitoyltransferase (SPT). Four missense mutations (C133W, C133Y, V144D and G387A) in SPTLC1 were reported to cause HSAN I. SPT catalyses the condensation of Serine and Palmitoyl-CoA, which is the first and rate-limiting step in the de novo synthesis of ceramides. Earlier studies showed that C133W and C133Y mutants have a reduced activity, whereas the impact of the V144D and G387A mutations on the human enzyme was not tested yet. In this paper, we show that none of the HSAN I mutations interferes with SPT complex formation. We demonstrate that also V144D has a reduced SPT activity, however to a lower extent than C133W and C133Y. In contrast, the G387A mutation showed no influence on SPT activity. Furthermore, the growth phenotype of LY-B cells--a SPTLC1 deficient CHO cell line--could be reversed by expressing either the wild-type SPTLC1 or the G387A mutant, but not the C133W mutant. This indicates that the G387A mutation is most likely not directly associated with HSAN I. These findings were genetically confirmed by the identification of a nuclear HSAN family which showed segregation of the G387A variant as a non synonymous SNP. PMID- 19132418 TI - A computational approach to studying monomer selectivity towards the template in an imprinted polymer. AB - A computational approach was proposed to study monomer-template interactions in a molecularly imprinted polymer (MIP) in order to gain insight at the molecular level into imprinting polymer selectivity, regarding complex formation between template and monomer at the pre-polymerisation step. This is the most important step in MIP preparation. In the present work, chlorphenamine (CPA), diphenhydramine (DHA) and methacrylic acid (MAA), were chosen as the template, non-template, and monomer, respectively. The attained complexes were optimised, and changes in the interaction energies, atomic charges, IR spectroscopy results, dipole moment, and polarisability were studied. The effects of solvent on template-monomer interactions were also investigated. According to a survey of the literature, this is the first work in which dipole moment and polarisability were used to predict the types of interactions existing in pre-polymerisation complexes. In addition, the density functional tight-binding (DFTB) method, an approximate version of the density functional theory (DFT) method that was extended to cover the London dispersion energy, was used to calculate the interaction energy. PMID- 19132420 TI - Are pelvic radiographs needed in assault victims? AB - To determine the incidence of acute pelvic fractures in patients with a history of being assaulted. Retrospective review by two musculoskeletal radiologists of 357 consecutive patients in a 12-month period, (308 males, 49 females). Two hundred seventy-six patients also had pelvic CT scans done at the same time. There were no acute pelvic fractures in this group. One patient had a 12th rib fracture and two other patients had a total of three lumbar spine transverse process fractures at the L1, L2, and L5 levels. Pelvic radiographs are not needed as part of standard protocols for patients who present with a history of being assaulted. PMID- 19132421 TI - Acute gastric outlet obstruction secondary to papillary serous adenocarcinoma of the endometrium with peritoneal psammomatous implants: a case report. AB - Endometrial carcinoma is the most common gynecologic cancer in the United States. Uterine papillary serous carcinoma comprises approximately 5-10% of endometrial carcinomas. This aggressive carcinoma typically occurs in older women, characteristically arising on atrophic endometrium. This frequently is associated with early extrauterine spread and, infrequently, with peritoneal dissemination. We present a case of acute gastric outlet obstruction secondary to papillary serous adenocarcinoma of the endometrium with diffuse peritoneal psammomatous implants. PMID- 19132422 TI - Musculoskeletal: what's different in children? Running and fell on the outstretched upper extremity. PMID- 19132423 TI - Air around the pulmonary vein: an unusual finding in a patient with blunt thoracic trauma. AB - Traumatic thoracic injuries and related complications have a mortality of 15.5 25%. We present a case of a 30-year-old with blunt thoracic trauma, massive edema of the lung, and laceration of the middle lobe, associated with air around the pulmonary vein, evaluated with multidetector computed tomography. To the best of our knowledge, air around the pulmonary vein has not been previously described in the literature. PMID- 19132424 TI - MR imaging of urgent inflammatory and infectious conditions affecting the soft tissues of the musculoskeletal system. AB - Soft tissue infections and inflammatory conditions of the musculoskeletal system are a group of disorders commonly seen by emergency room physicians and radiologists. Many of these entities can either be limb- or life-threatening. Magnetic resonance imaging is currently the best imaging modality to evaluate these conditions. In this review, the characteristic imaging findings of cellulitis, abscess formation, necrotizing fasciitis, pyomyositis, diabetic ischemic infarction, acute and exertional compartment syndromes, and rhabdomyolysis will be emphasized as well as imaging factors that can help to differentiate these disorders. PMID- 19132425 TI - Accuracy of CT angiography for the diagnosis of vascular abnormalities causing intraparenchymal hemorrhage in young patients. AB - The objective of this study is to measure the accuracy of multidetector CT angiography (MD CTA) in the detection of vascular abnormalities in patients 60 years). RESULTS: In the Emergency Department two patients were comatose and required intubation. Another patient was intubated within the first 24 h because of ARDS. At presentation, all patients but two had a PaO(2)/FiO(2) < 300, while ARDS was present in 17 and acute lung injury in 15 cases. The nine remaining hypoxemic patients had either focal infiltrates or a negative chest X-ray. Superimposed pneumonia was observed in four patients and resulted in a protracted hospital stay. Improvement of lung injury was rapid in most cases: by day 4 resolution of hypoxemia was observed in 33/43 (76.7%) of the cases and resolution of radiographic findings in 66.6%. Duration of hospitalization varied from 2 to 14 days (mean = 5.2 +/- 0.5 days). One patient with coma died due to ventilator-associated pneumonia (mortality = 2.3%). CONCLUSION: Respiratory manifestations of near-drowning in adult immersion victims are often severe. Nevertheless, in noncomatose patients at least, intubation can often be avoided and quick improvement is the rule while a good outcome is usually expected even in elderly patients. PMID- 19132446 TI - Clinical considerations on sentinel node biopsy in melanoma from an Italian multicentric study on 1,313 patients (SOLISM-IMI). AB - BACKGROUND: Although widely used for the management of patients with cutaneous melanoma, the sentinel lymph node (SLN) biopsy (SNB) procedure raises several issues. This study was designed to investigate: the predictive factors of SLN status, the false-negative (FN) rate, and patients' prognosis after SNB. PATIENTS AND METHODS: This is an observational, prospective study conducted on a large series of consecutive patients (n = 1,313) enrolled by 23 Italian centers from 2000 through 2002. A commonly shared protocol was adopted for the SNB surgical procedure and the SLN pathological examination. RESULTS: The SLN positive and false-negative (FN) rates were 16.9% and 14.4%, respectively (median follow-up, 4.5 years). At multivariable logistic regression analysis, the frequency of positive SLN increased with increasing Breslow thickness (p < 0.0001) and decreased in patients with melanoma regression (p = 0.024). At the multivariable Cox regression analysis, SLN status was the most important prognostic factor (hazards ratio (HR) = 3.08) for overall survival; the other statistically significant factors were sex, age, Breslow thickness, and Clark's level. Considering SLN and NSLN status, including FN cases, we identified four groups of patients with different prognoses. The 5-year overall survival of patients with positive SLNs was 71.3% in those with negative nonsentinel lymph nodes (NSLNs) and 50.4% if NSLNs were positive. CONCLUSIONS: Regression in the primary melanoma seems to be a protective factor from metastasis in the SLN. When correctly calculated, the SNB FN rate is 15-20%. Furthermore, the SNB is important to more precisely assess the prognosis of patients with melanoma. PMID- 19132447 TI - Preoperative chemotherapy decreases the need for re-excision of breast cancers between 2 and 4 cm diameter. AB - INTRODUCTION: It is accepted that preoperative chemotherapy can result in increased breast preservation for breast cancers greater than 4 cm. The benefits of preoperative chemotherapy are less clear, however, for patients who present with smaller tumors and are already candidates for breast-preserving surgery. The goal of this study is to assess the effect of preoperative chemotherapy on breast cancers between 2 and 4 cm diameter. METHODS: A retrospective chart review was conducted of patients diagnosed with new breast cancer at the Yale-New Haven Breast Center for the years 2002-2007. Patients were included in the study if their breast cancer was between 2 and 4 cm and their initial surgical treatment had been completed. Patients with distant metastases were excluded. RESULTS: There were 156 new cancers that met study requirements. Forty-seven patients underwent preoperative chemotherapy, and 109 patients had their surgery first, usually followed by chemotherapy. Initial surgery was lumpectomy for 31 out of 47 patients (66%) in the preoperative chemotherapy group compared with 62 out of 109 patients (57%) in the surgery group. For patients with lumpectomies, 2 out of 31 patients (6%) in the preoperative group had positive margins and required re excision compared with 20 out of 62 patients (37%) in the surgery-first group (P<0.01). CONCLUSIONS: We conclude that, for tumors between 2 and 4 cm, preoperative chemotherapy is associated with a significantly decreased rate of re excision following lumpectomy. This not only results in fewer mastectomies, but also avoids the morbidity and inferior cosmetic results associated with a re excision lumpectomy. PMID- 19132448 TI - Dynamic monitoring of apoptosis in chemotherapies with multiple fluorescence reporters. AB - PURPOSE: The aim of the study is to dynamically and non-invasively monitor the temporal relationship among caspase-3, BID, and cytochrome c in chemotherapy. PROCEDURES: ASTC-a-1 cells expressing the corresponding fluorescence reporters were treated with Taxol or cisplatin and imaged using FRET and fluorescence overlapping technique. Western blot was performed to validate the fluorescence analysis. RESULTS: In fluorescence imaging analysis, Taxol-induced apoptosis showed caspase-3 activation (13 h 50 min) was prior to BID cleavage (15 h 10 min) and subsequent significant cytochrome c release (17-18 h 20 min), whereas the cisplatin-induced apoptosis showed BID cleavage (5 h 40 min) and significant cytochrome c release (7-8 h 20 min) were prior to caspase-3 activation (14 h 20 min). Western blot further validated the results above. CONCLUSIONS: The new approach successfully reveals the difference in temporal signaling apoptosis events between Taxol and cisplatin. It may help us come to a better understanding of the detailed mechanisms in chemotherapeutic-agents-induced apoptosis. PMID- 19132449 TI - Metabolic changes in the rodent brain after acute administration of salvinorin A. AB - PURPOSE: Salvinorin A (SA) is a potent and highly selective kappa-opioid receptor (KOR) agonist with rapid kinetics and commensurate behavioral effects; however, brain regions associated with these effects have not been determined. PROCEDURES: Freely moving adult male rats were given SA intraperitoneally during uptake and trapping of the brain metabolic radiotracer, 2-deoxy-2-[F-18]fluoro-D: -glucose (FDG), followed by image acquisition in a dedicated animal positron emission tomography (PET) system. Age-matched control animals received vehicle treatment. Animal behavior during FDG uptake was recorded digitally and later analyzed for locomotion. Group differences in regional FDG uptake normalized to whole brain were determined using Statistical Parametric Mapping (SPM) and verified by region of interest (ROI) analysis. RESULTS: SA-treated animals demonstrated significant increases in FDG uptake compared to controls in several brain regions associated with the distribution of KOR such as the periaqueductal grey, bed nucleus of the stria terminalis and the cerebellar vermis, as well as in the hypothalamus. Significant bilateral activations were also observed in the auditory, sensory, and frontal cortices. Regional decreases in metabolic demand were observed bilaterally in the dorsolateral striatum and hippocampus. Locomotor activity did not differ between SA and vehicle during FDG uptake. CONCLUSIONS: We have provided the first extensive maps of cerebral metabolic activation due to the potent kappa-opioid agonist, salvinorin A. A major finding from our small animal PET studies using FDG was that neural circuits affected by SA may not be limited to direct activation or inhibition of kappa-receptor-expressing cells. Instead, salvinorin A may trigger brain circuits that mediate the effects of the drug on cognition, mood, fear and anxiety, and motor output. PMID- 19132450 TI - Local treatment for recurrent colorectal hepatic metastases after partial hepatectomy. AB - OBJECTIVE: The objective of the study was to identify patients who may benefit from local treatment in recurrent colorectal liver metastases. MATERIALS AND METHODS: A total of 51 consecutive patients were treated for hepatic recurrence(s) after an initial partial hepatic resection. Surgery was considered as the primary treatment option for eligible patients. Patients with a small liver remnant after major hepatectomy were treated with radiofrequency ablation (RFA) or stereotactic body radiation therapy (SRx). SRx was given as an outpatient, emerging local treatment option for patients with intra-hepatic recurrences not eligible for surgery or RFA. Partial liver resection was performed in 36 patients (70%), RFA in ten patients (20%), and SRx in five patients (10%). RESULTS: Median hospital stay was 7 (range, 3-62) days with a morbidity of 16% without in-hospital death. None of the patients received adjuvant chemotherapy. There was no difference in recurrence or survival between the three treatment modalities. Overall 5-year survival was 35% with an estimated median survival of 37 months. Patients with a disease-free interval between first hepatectomy and hepatic recurrence less than 6 months did not survive 3 years. CONCLUSIONS: Resection, RFA, and SRx can be performed safely in patients with recurrent colorectal liver metastases and offer a survival that seems comparable to primary liver resections of colorectal liver metastases. PMID- 19132451 TI - Colectomy in patients with acute colitis: a systematic review. AB - BACKGROUND: For patients with acute colitis, the decision when and how to operate is difficult in most cases. It was the aim of this systematic review to analyze early mortality and morbidity of colectomy for severe acute colitis in order to identify opportunities to improve perioperative treatment and outcome. METHODS: A systematic review of the available literature in the Medline and PubMed databases from 1975 to 2007 was performed. All articles were assessed methodologically; the articles of poor methodological quality were excluded. Articles on laparoscopic colectomy for acute colitis were analyzed separately. RESULTS: In total, 29 studies met the criteria for the systematic review, describing a total of 2,714 patients, 1,257 of whom were operated on in an acute setting, i.e., urgent or emergency colectomy. Reported in-hospital mortality was 8.0%; the 30-day mortality was 5.2%. Morbidity was 50.8%. The majority of complications were of infectious and thromboembolic nature. Over the last three decades, there was a shift in indications from toxic megacolon, from 71.1% in 1975-1984 to 21.6% in 1995-2005, to severe acute colitis not responding to conservative treatment, from 16.5% in 1975-1984 to 58.1% in 1995-2007. Mortality decreased from 10.0% to 1.8%. Morbidity remained high, exceeding 40% in the last decade. Mortality after laparoscopic surgery was 0.6%. Complication rate varies from 16-37%. CONCLUSION: Colectomy for acute colitis is complicated by considerable morbidity. The incidence of adverse outcome has substantially decreased over the last three decades, but further improvements are still required. The retrospective nature of the included studies allows for a considerable degree of selection bias that limits robust and clinically sound conclusions about both conventional and laparoscopic surgery. PMID- 19132452 TI - Suspension suture augmentation for repair of coracoclavicular ligament disruptions. AB - Surgical reconstruction of the coracoclavicular ligament is a fundamental part of management of high-grade acromioclavicular dislocations and Type II lateral third clavicular fractures. However, no single surgical procedure is fully satisfactory because of failure or complications. We present an alternative coracoclavicular stabilization technique, which avoids the use of hardware or tendon graft, that was used in 10 consecutive patients with complete coracoclavicular ligament disruptions. These patients were followed for a minimum of 14 months (average, 34.8 months; range, 14-55 months). At the final followup, functional outcome measurement instruments (University of California-Los Angeles shoulder rating system and Western Ontario Shoulder Instability Index) and radiographic analysis were adopted as the main outcome measures of shoulder function. The mean University of California-Los Angeles shoulder rating score and the mean Western Ontario Shoulder Instability Index aggregation score at 12 months after surgery were 33.8 (95% confidence interval, 32.8-34.8) and 93.4 (95% confidence interval, 88.2-98.6), respectively. The radiographic analysis revealed all patients had maintained reduction on radiographs at the final followup. These preliminary results suggest that this simple technique can achieve stable coracoclavicular reconstruction and facilitate healing of the repaired ligaments or fractures. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19132453 TI - Femoroacetabular impingement: current status of diagnosis and treatment: Marius Nygaard Smith-Petersen, 1886-1953. AB - This biographical sketch of M. N. Smith-Petersen corresponds to the historic text, The Classic: Treatment of Malum Coxae Senilis, Old Slipped Upper Femoral Epiphysis, Intrapelvic Protrusion of the Acetabulum, and Coxa Plana by Means of Acetabuloplasty, available at DOI 10.1007/s11999-008-0670-0 . PMID- 19132456 TI - Perceived barriers to paracetamol (acetaminophen) prescribing, especially following rofecoxib withdrawal from the market. AB - Cyclooxygenase-2 inhibitor (COX-2) inhibitors were publicly subsidized in Australia for osteoarthritis. However, guidelines still recommended paracetamol as first choice therapy. When rofecoxib was withdrawn in 2004, paracetamol should have been offered as replacement. However, dispensing data indicate no increase in paracetamol use. The objective of this study was to gain understanding about barriers to paracetamol use and to identify what choices consumers were offered after rofecoxib withdrawal. We conducted two focus groups (consumers and pharmacists) and 15 semi-structured interviews (seven with patients taking rofecoxib at the time it was withdrawn in Australia, four with pharmacists, and four with general practitioners). Familiarity with and use of paracetamol, perceived strengths and weaknesses of paracetamol for chronic pain, and choices given about therapy changes were investigated. All interviews and focus groups were recorded, transcribed verbatim, and thematically analyzed. Consumers reported that transfer of information on their medicines was limited or absent. They perceived that their knowledge about COX-2 inhibitor safety and/or appropriate use of paracetamol was lacking. Pharmacists agreed that several factors were relevant concerning paracetamol and COX-2 inhibitor use, including lack of counseling and information for consumers. Not personalizing prescribing to elderly patients was identified as a weakness. Consumers who had received rofecoxib were divided about their perceptions of the efficacy of paracetamol. It appears that when rofecoxib was withdrawn, they were not offered an opportunity to try paracetamol. Consumers in this study appeared to have poor knowledge about the opportunity to effectively use paracetamol. Consumers did not remember being given the choice to use paracetamol as regular treatment for chronic pain. Pharmacists and doctors did not appear to be discussing options for pain control well with consumers and had mismatched perceptions with consumers about paracetamol. An educational intervention to encourage more rational use of paracetamol is now being planned to provide consumers with more knowledge about paracetamol effective use. PMID- 19132457 TI - Acute eosinophilic pneumonia is a non-infectious lung complication after allogeneic hematopoietic stem cell transplantation. AB - Acute eosinophilic pneumonia (AEP) is an acute febrile illness with respiratory impairment, diffuse pulmonary infiltrates, and eosinophilia in bronchoalveolar lavage (BAL) fluid. We report an adult male who developed severe cough and dyspnea with slight fever on day 78 after allogeneic hematopoietic stem transplantation. The symptoms coexisted with skin and gut GVHD. The imaging test demonstrated interstitial infiltrates and BAL analysis revealed marked increase of eosinophils and no sign of infection. We made a diagnosis of AEP and steroid was started. AEP remitted with other GVHD symptoms but exacerbated partially when steroid was decreased. This case suggests a potential link between AEP and GVHD. PMID- 19132458 TI - Ruptured intramural intestinal hematoma in an adolescent patient with severe hemophilia A. AB - We report on a 17-year-old patient with severe hemophilia A without inhibitors who developed abdominal bleeding after an episode of severe cough. Abdominal ultrasound showed intramural intestinal hematoma as well as large amount of peritoneal fluid appearing as blood and right hematocele. Abdominal CT revealed markedly thickened intestinal wall in sigmoidal region. Patient was managed with replacement therapy as well as peritoneal drainage with favorable outcome. This is the first report on a hematoperitoneum in a hemophiliac due to ruptured intramural sigmoidal hematoma. PMID- 19132459 TI - What is the real contribution of extrahepatic cells to liver regeneration? AB - Extrahepatic cells, especially bone marrow (BM) cells, might contribute to liver repair, but recent published evidence suggests that they do not play a role in the normally regenerating liver. The mechanism by which extrahepatic cells express a liver-specific function in the liver, whether by transdifferentiation or by cell fusion, remains unclear. In this review, we investigate the status of findings on this controversial subject and summarize the recent research. PMID- 19132460 TI - Clinical and pathological features of intracystic papillary carcinoma of the breast. AB - PURPOSE: To evaluate the clinicopathological features of intracystic papillary carcinoma (ICPC), which have not been established given its rarity and lack of standard diagnostic criteria. METHODS: We reviewed the clinicopathological findings and treatment outcomes of 14 patients with ICPC diagnosed between 2002 and 2006. RESULTS: Intracystic papillary carcinoma was diagnosed by fine-needle aspiration biopsy in three patients and by core-needle biopsy in six patients. A preoperative diagnosis was not made in five patients. Three patients underwent magnetic resonance imaging preoperatively, which helped to differentiate benign tumors and maintain free surgical margins. The final pathological diagnosis was invasive carcinoma in 2 (14.2%) of the 14 patients. The patients were followed up for 1-72 months, during which time only one died, of a cancer-unrelated cause. CONCLUSION: Our results show that ICPC is more difficult to diagnose than common breast cancer preoperatively. Excisional biopsy was necessary when fine-needle aspiration and core-needle biopsy could not provide a diagnosis. Magnetic resonance imaging is helpful to differentiate a benign tumor from invasive disease. PMID- 19132461 TI - Transcutaneous carbon dioxide tension for the evaluation of limb ischemia. AB - PURPOSE: Understanding the hemodynamics of critical limb ischemia caused by chronic peripheral arterial occlusive disease is important to evaluate its severity and the efficacy of treatment. We investigated the usefulness of transcutaneous carbon dioxide tension (tcPCO(2)) measurement for evaluating ischemic limbs, in conjunction with the measurement of ankle pressure (AP), toe pressure (TP), skin perfusion pressure (SPP), and transcutaneous oxygen tension (tcPO(2)). METHODS: We measured tcPCO(2) in the dorsum of the foot in 158 patients (304 limbs) with arteriosclerosis obliterans. RESULTS: The tcPCO(2) in normal limbs without any clinical sign or abnormal noninvasive measurement was 43.7 +/- 3.7 mmHg; that in noncritical ischemic limbs was 45.5 +/- 9.0 mmHg, which was not significantly different from that in the normal limbs; and that in critically ischemic limbs was 87.6 +/- 35.5 mmHg, which was significantly different from that in the normal limbs. All limbs with a tcPCO(2) of 100 mmHg or higher, indicative of critical ischemia, had a tcPCO(2) of less than 100 mmHg after revascularization. CONCLUSION: We found tcPCO(2) to be a useful measurement for diagnosing the severity of limb ischemia, and for evaluating the effect of treatment, especially in patients with critically ischemic limbs. PMID- 19132463 TI - Stent placement for superficial femoral arterial occlusive disease in high-risk patients: preliminary results. AB - PURPOSE: To report the preliminary results of primary stenting of the superficial femoral artery (SFA) in high-risk patients with symptomatic SFA occlusive disease. METHODS: Between February 2005 and April 2007, a collective 30 lower limbs in 28 high-risk patients with SFA occlusive disease were treated by primary stenting. Hemodynamic improvement was assessed by ankle brachial pressure index (ABI), before and after the procedure. Primary and primary-assisted patency and limb salvage were measured in concordance with the Society for Vascular Surgery guidelines. RESULTS: The technical success rate was 97%. The average ABI before and after stent placement increased significantly, from 0.41 +/- 0.33 to 0.82 +/- 0.20 (P < 0.001). The primary and primary-assisted patency rates were 86% and 90% at 6 months, 86% and 90% at 12 months, and 68% and 90% at 24 months, respectively. The limb salvage rate was 92% at 6, 12, and 24 months and the survival rate was 95% at 6, 12, and 24 months. CONCLUSION: Our results suggest that the primary stenting of SFA lesions is a feasible, safe, and effective procedure for high-risk patients with SFA occlusive disease. PMID- 19132462 TI - Quantitative assessment of therapeutic effects in the critically ischemic limb using (99m)Tc-diethylene-triamine-pentaacetic acid human serum albumin. AB - PURPOSE: To investigate the reliability and limitations of a new radioisotope method using (99m)Tc-diethylenetriamine-pentaacetic acid human serum albumin (Tc 99m-DTPA-HSA) and to evaluate the diagnostic ability of isotope infusion for assessing hemodynamic changes in the foot before and after treatment. METHODS: Hemodynamic changes before and after treatment were assessed in 21 limbs with ulcer or gangrene, by analyzing changes in the time-activity curve, the uptake ratio, and the values obtained with noninvasive techniques. RESULTS: There were significant differences between each pair of the three types of time-activity curve and their uptake counts. The uptake ratio was correlated with ankle blood pressure (ABP) and toe blood pressure (TBP), but not with transcutaneous oxygen pressure (tcPO(2)) or skin perfusion pressure (SPP). The hemodynamic change induced by pharmacotherapy was subtle, but that induced by arterial reconstruction was remarkable. Although there was not always a good correlation between the degree of hemodynamic change and the clinical outcome in limbs treated with pharmacotherapy, the hemodynamic change was quantitatively assessed. CONCLUSION: Our study suggests that this isotope technique is a useful quantitative method to evaluate hemodynamic change from a different perspective to conventional noninvasive methods. PMID- 19132464 TI - Chemotherapy for small-bowel Adenocarcinoma at a single institution. AB - PURPOSE: Small-bowel adenocarcinoma (SBA) is rare. No standard chemotherapy for this type of cancer has yet been established. At Cancer Institute Hospital (CIH), the chemotherapy regimen used for colorectal cancer is initially used for patients with SBA, followed by that used for gastric cancer. METHODS: Patients with advanced or recurrent SBA who had been treated with chemotherapy in CIH were retrospectively analyzed. The first-line treatments were fluoropyrimidines used alone or in combination with other drugs, such as 5-fluorouracil plus leucovorin (FL), UFT-E, or TS-1. The second-line treatment was irinotecan (CPT-11) monotherapy. RESULTS: Fluoropyrimidine-based regimens, mainly FL, were used for 10 patients. Seven patients received the second-line CPT-11 regimen. Disease control was seen in five patients (50%) with the first-line chemotherapy and in three (43%) with the second-line. The median overall survival time was 12 months (range 3-39). The treatments were generally tolerated. Gastrointestinal symptoms were the most common adverse effects. CONCLUSIONS: Fluoropyrimidines as the first line and CPT-11 as the second-line chemotherapy yielded low response, although the adverse effects were mild. The FOLFOX and FOLFIRI regimens such as those used for metastatic colorectal cancer are potential alternative strategies. Extensive trials are needed to develop standard chemotherapy with new drugs. PMID- 19132465 TI - Roux-en-Y reconstruction using staplers during pancreaticoduodenectomy: results of a prospective preliminary study. AB - PURPOSE: The aim of this study was to reveal the utility of alimentary reconstruction using staplers during pancreaticoduodenectomy (PD), focusing on the occurrence of delayed gastric emptying. METHODS: Between 2003 and 2007, 72 PDs with alimentary reconstruction were performed by a single surgeon. Since August 2006, the new Roux-en-Y reconstruction methods using staplers were applied in 26 of the patients. We compared their clinical outcomes with those of the 46 patients who underwent PD using the conventional hand-sewn reconstruction methods. RESULTS: The results of upper gastrointestinal study showed improvement within 10 postoperative days (PODs; P = 0.03): the patients resumed eating their regular diet sooner (13 vs 6 days, P < 0.001), and both the incidence of delayed gastric emptying (43% vs 19%, P = 0.04) and the hospital stay (27 vs 21 days, P = 0.008) were reduced significantly in patients with stapled reconstruction. Despite the fact that operative costs were significantly higher for patients with stapled reconstruction (P = 0.009), hospital costs were significantly lower (P = 0.049) for those who underwent the conventional method. CONCLUSIONS: Our retrospective analysis shows that stapled reconstructions might reduce the incidence of delayed gastric emptying; however, further study will be necessary to evaluate the utility of this new method. PMID- 19132466 TI - Effects of oxaliplatin and 5-fluorouracil on the healing of colon anastomoses. AB - PURPOSE: Oxaliplatin (OX) and 5-fluorouracil (5-FU) are the most widely used chemotherapeutic agents in the adjuvant treatment of colon cancer. Although the early initiation of adjuvant chemotherapy can improve the outcome of surgery, it carries potentially fatal risks. This experimental study investigates the effects of 5-FU and OX on colon anastomoses. METHODS: We used 60 rats, divided into six groups. After being subjected to bowel resection and anastomosis, the rats were given 5-FU on days 1-3, or OX 130 mg/m(2) on days 1 or 5, or 5% dextrose as a control. The bursting pressures and hydroxyproline content of the anastomoses were measured, and complications and adhesions were recorded. RESULTS: There were no major complications in the treatment groups. The bursting pressures of the 5 FU group were significantly lower than those of the control and OX groups. The bursting pressures of the OX groups were not significantly different from those of the control groups. The hydroxyproline levels of the rats treated with OX on day 1 were significantly lower than those of the rats treated with OX on day 5 and the 5-FU groups. CONCLUSION: Oxaliplatin and 5-FU did not compromise wound healing of the colon significantly. Our results indicate that OX is less detrimental to the healing of colonic anastomoses, when administered on days 1 and 5 after resection, than 5-FU. PMID- 19132467 TI - Solitary cranial metastasis of thyroid carcinoma 13 years after primary surgery: report of a case. AB - Thyroid cancers rarely metastasize to the brain: the incidence of brain metastasis of differential thyroid carcinomas has been reported at only 0.69% 1.3%. We report a case of a solitary brain metastasis from thyroid follicular carcinoma found 13 years after primary surgery. A 73-year-old woman was admitted to our hospital for investigation of headaches, double vision, and facial nerve palsy, 13 years after a subtotal thyroidectomy for thyroid follicular carcinoma. Magnetic resonance imaging (MRI) showed a solitary mass in the midbrain associated with a focal osteolytic lesion of the temporal bone. Further radiological examinations revealed no other mass lesion. After making a provisional preoperative diagnosis of a primary brain tumor, we performed subtotal removal of the tumor. Histopathological examination confirmed metastasis of thyroid follicular carcinoma. Thus, the patient underwent completion thyroidectomy followed by internal radiotherapy. She has remained well in the 4 years since her operation, without any signs of further recurrence. PMID- 19132468 TI - Osteoclast-like giant cell tumor arising in the soft tissue of the breast: report of a case. AB - Extraosseous manifestations of osteoclast-like giant cell tumors (OGCTs) in soft tissue are unusual, especially in the breast. However, multinucleated osteoclast like giant cells have been described in association with epithelial malignancy, as a variant of breast carcinoma. We report a case of OGCT of the soft tissue of the breast, not associated with epithelial elements. To the best of our knowledge, this is only the second such case reported. PMID- 19132469 TI - Spontaneous rupture of the spleen in type IV Ehlers-Danlos syndrome: report of a case. AB - The vascular type of Ehlers-Danlos syndrome, type IV, is associated with severe complications, including arterial rupture and visceral perforation. However, to our knowledge, there has been only one previous report of splenic rupture caused by a spontaneous hemorrhage in type IV Ehlers-Danlos syndrome. We report another case of this uncommon complication, occurring in a 35-year-old woman who presented after the sudden onset of acute abdominal pain. Patients should be stabilized quickly in the intensive care unit and the most timesaving surgical techniques used. Moreover, tissues must be handled with great care intraoperatively in view of their extreme fragility. Despite prompt and appropriate treatment, the prognosis is often dismal. PMID- 19132470 TI - Reconstructive surgery for a fusiform common celiomesenteric trunk aneurysm and coexistent abdominal aortic aneurysm: report of a case. AB - A celiomesenteric trunk (CMT) accounts for fewer than 1% of all visceral artery anomalies. Aneurysms involving a CMT are exceptionally rare: our search of the literature found only eight cases reported. We treated a 73-year-old man with both a fusiform CMT aneurysm and an abdominal aortic aneurysm (AAA). The celiac artery and superior mesenteric artery were reconstructed by performing a retrograde prosthetic-graft bypass originating from the right limb of the bifurcated aortic graft used to repair the AAA. To our knowledge, this is the first report of successful surgical treatment of a CMT aneurysm and coexistent AAA. PMID- 19132472 TI - Primary omental torsion: report of two cases. AB - Two patients with omental torsion, who presented with acute abdomen, are reported. The first case, a 14-yearold boy, was admitted for acute appendicitis. During surgery, omental torsion on the long axis was diagnosed and the involved omentum was resected. The patient had not experienced any previous abdominal surgery. The second case, a 49-year-old man, was admitted with symptoms of acute abdomen. Similarly, a laparotomy revealed omental torsion around the long axis. This patient also had no history of previous abdominal surgery. The involved portion of the omentum was also resected. Omental torsion is a rare cause of acute abdomen in children and adults, who may present with various signs and symptoms mimicking other etiologies of acute abdomen. A preoperative diagnosis may therefore be difficult and it usually can only be established during surgery. PMID- 19132471 TI - Proctocolectomy for colon cancer associated with ulcerative colitis a few months after living donor liver transplantation for primary sclerosing cholangitis: report of a case. AB - Colorectal cancer (CRC) frequently develops in patients with ulcerative colitis (UC). We report a case of CRC treated successfully by proctocolectomy 8 months after living donor liver transplantation (LDLT) for primary sclerosing cholangitis (PSC). The lesion was detected early, probably as a result of colonoscopic surveillance after LDLT. Thus, patients with a long history of UC, who undergo LDLT for PSC, should be followed up with regular surveillance colonoscopy. Moreover, surgery, such as radical resection of the colon and rectum should be performed without delay, even shortly after LDLT. To our knowledge, this is the first report of a patient undergoing proctocolectomy after LDLT. PMID- 19132473 TI - Successful utilization of coronary covered stents to treat a common hepatic artery pseudoaneurysm secondary to pancreatic fistula after Whipple's procedure: report of a case. AB - A 73-year-old man underwent a pylorus-preserving Whipple's procedure for distal cholangiocarcinoma. His postoperative course was complicated by the formation of a pancreatic fistula, which was initially managed conservatively. On postoperative day (POD) 86, he lost 100 ml of blood from the site of the pancreatic fistula. Contrast-enhanced computed tomography (CT) showed a pseudoaneurysm, 12 mm in diameter, in the common hepatic artery. The diameter of the pseudoaneurysm increased to 15 mm on POD 89, so we implanted coronary covered stents to prevent massive bleeding from rupture and to retain hepatic arterial flow. Six days after implantation, computed tomography findings confirmed a thrombosed pseudoaneurysm as well as patent hepatic arterial flow. Follow-up CT 18 months after surgery showed patent hepatic arterial flow. There have been no signs of rebleeding or abnormal liver function. PMID- 19132474 TI - Abdominal wall recurrence of Hilar bile duct cancer 12 years after a curative resection: report of a case. AB - A 74-year-old woman was diagnosed with hilar bile duct cancer, and underwent a curative resection of the bile duct and the left and caudate lobes of the liver in 1995. Ten years later (April 2005), she noted a small mass in the abdominal wall. The mass slowly enlarged to reach 4 cm in diameter by January 2007. With a diagnosis of a possible recurrence of bile duct cancer, a laparotomy was thus performed. The abdominal wall tumor was buried in the rectus abdominis muscle and was tightly attached to the ileum. The lesion was resected en bloc with the associated rectus muscle and ileocecal region. A histopathological examination of the resected specimen revealed tubular adenocarcinoma that closely resembled the original primary bile duct cancer. In addition, the immunohistochemical staining pattern of the abdominal tumor was identical to that of the original bile duct cancer. This indicated that the abdominal tumor represented a local recurrence (probably due to peritoneal implantation) at 12 years after the resection of the hilar bile duct cancer. This case emphasizes that long-time surveillance is required for patients with bile duct cancer, even if they have survived without recurrence for more than 5 years after a curative resection. PMID- 19132475 TI - Type VI biliary cyst: report of a case. AB - An isolated cyst of the cystic duct is an extremely rare lesion. Only single case reports are documented in the literature. The most accepted classification system of biliary cysts, the Todani classification, does not include this lesion. We report a case of isolated cyst of the cystic duct. The initial referral was for evaluation of a gallbladder mass discovered during evaluation of abdominal pain. Preoperative diagnosis was challenging as multiple imaging studies were unable to differentiate this lesion from a choledochal cyst. Surgical planning thus included cyst excision and biliary reconstruction. Operative exploration revealed a type VI biliary cyst and cholecystectomy with cystic duct ligation near the common bile duct was curative. PMID- 19132476 TI - Torsion of an accessory lobe of the liver in a child: report of a case. AB - We report a case of torsion of an accessory lobe of the liver (ALL) in a 14-year old girl. The patient was admitted for acute abdominal pain and laparotomy revealed an ALL, the pedicle of which was elongated with 180 degrees torsion. The diagnosis was not made preoperatively because of the rarity of this condition. However, a close relationship between omphalocele repair and the development of ALL has been reported; thus, a history of omphalocele repair should alert the doctor to the possibility of this condition, which could be suggested by imaging findings preoperatively. Torsion of an ALL should be included in the differential diagnosis of a patient with a history of omphalocele, who presents with acute abdominal symptoms. PMID- 19132477 TI - A new procedure with stomach-lifting techniques to simplify laparoscopy-assisted distal gastrectomy and extraperigastric lymph node dissection for gastric cancer. AB - Laparoscopy-assisted gastrectomy with extraperigastric lymph node dissection for gastric cancer is a complicated procedure that generally requires advanced laparoscopic surgical skill. We devised a simplified but effective laparoscopic procedure that provides a better visual field to perform safe lymph node dissection more quickly. First, a mini-laparotomy is done and a clear visual field is created by pulling a mini-retractor to the right or left. The laparoscopic procedure is made easier and safer by taping the stomach body, and by using the fringe of an abdominal wall sealing device (Lapdisk) placed behind the stomach, and a scope holder for the snake-retractor. The lymph nodes along the common hepatic vessels, left gastric vessels, and celiac artery (extraperigastric lymph nodes) are then dissected laparoscopically. The suprapyloric and infrapyloric lymph nodes are dissected through the mini laparotomy incision and gastroduodenostomy is done using an anastomotic device. We performed laparoscopy-assisted distal gastrectomy (LADG) in 70 patients with gastric carcinomas located in the distal stomach (mean body mass index: 24.3). The mean operating time was 170 min and blood loss was minimal. All patients recovered well with minimal pain and good postoperative quality of life. We conclude that our simple and practical procedure for LADG with extraperigastric lymph node dissection can be performed safely and easily. PMID- 19132478 TI - Ludwik Rydygier--contributor to modern surgery. AB - Modern surgery is usually considered to have begun in nineteenth century Europe. One of the most famous contributors to gastric cancer surgery was the Polish surgeon Ludwik Rydygier, born in 1850. He initiated new methods in several fields, such as gastrointestinal surgery, orthopedics, gynecology, and urology. He was the second surgeon in the world to perform an antral resection, which he carried out on November 16, 1880. The patient, a 64-year-old man, suffered from pyloric cancer and died 12 hours after the procedure as a result of postoperative shock. The next pyloric resection was performed by Billroth in Vienna in 1881. In the nineteenth century few gastric resections were performed for peptic ulcer. The first successful antral resection for gastric ulcer penetrating to the pancreas was also performed by Ludwik Rydygier, in 1881. For many years Rydygier advocated resection in the treatment of gastric ulcers, although it was considered too dangerous for benign disease. He eventually proposed four indications for gastric resection: antral cancer, gastric ulcer, perforated gastric ulcer, and bleeding ulcers. Another operation performed for the first time by Ludwik Rydygier was gastroenterostomy, in a patient with a duodenal ulcer. In the following years other types of partial gastric resection and total gastrectomy were introduced. In 1992 the Ludwik Rydygier Association was founded in Krakow to commemorate the achievements of and pay tribute to this great surgeon. The Eighth International Gastric Cancer Congress will take place in 2009 in Krakow, where Ludwik Rydygier built a new surgical clinic in 1889. PMID- 19132479 TI - Disseminated cancer cells in the peritoneal cavity: what can we do when we detect them? PMID- 19132480 TI - Evaluation of lymph node metastases in gastric cancer using magnetic resonance imaging with ultrasmall superparamagnetic iron oxide (USPIO): diagnostic performance in post-contrast images using new diagnostic criteria. AB - BACKGROUND: We assessed the value of magnetic resonance imaging (MRI), using ultrasmall superparamagnetic iron oxide (USPIO) with new diagnostic criteria, in the evaluation of regional lymph node metastases in gastric cancer. METHODS: Thirty-one patients with gastric cancer were enrolled. 1000 lymph nodes were dissected during surgery, and of these, 519 nodes (51.9%) were identified by currently used MRI imaging analysis. We evaluated lymph nodes on USPIO-post contrast T2*-weighted images using the following two criteria: (1) we diagnosed the nodes on T2*-weighted images according to conventional criteria, where a node having an overall low signal intensity (pattern A) was nonmetastatic, while a node having partial (pattern B) or overall (pattern C) high signal intensity was metastatic; (2) we subdivided pattern B nodes on T1-weighted images using the new criteria, in which a node for which the high-intensity area on T2*-weighted images was not defined as adipose tissue on T1-weighted images (pattern B1) was metastatic, while a node for which the high-intensity area was defined as adipose tissue (pattern B2) was nonmetastatic. RESULTS: (1) The results using the conventional criteria were 96.2% sensitivity, 92.5% specificity, 76.3% positive predictive value (PPV), 99.0% negative predictive value (NPV), and 93.3% accuracy. (2) The results using the new criteria were 96.2% sensitivity, 98.3% specificity, 90.1% PPV, 99.0% NPV, and 97.1% accuracy. CONCLUSION: The assessment of lymph node metastases from USPIO-post-contrast MRI alone using the new criteria was useful in the diagnosis of regional lymph node metastases in gastric cancer. PMID- 19132481 TI - Combination chemotherapy with cisplatin and irinotecan in patients with adenocarcinoma of the small intestine. AB - BACKGROUND: Small-bowel adenocarcinoma (SBA) is a rare tumor that has a poor response to chemotherapy and a poor prognosis. Treatment strategies for SBA have not been clearly established. METHODS: All patients with SBA treated using a combination of cisplatin and irinotecan (IP) as first-line chemotherapy at the National Cancer Center Hospital in Japan between January 1999 and February 2007 were studied retrospectively. RESULTS: Eight patients received IP as first-line chemotherapy. The median follow-up was 9.5 months (range, 4.2-37.5 months). The median number of cycles of IP was three (range, 1-5). The overall response rate (complete or partial response) was 12.5% (complete response, n = 0; partial response, n = 1). The disease control rate (complete or partial response or stable disease) was 75%. The median time to treatment failure was 4.5 months (95% confidence interval, 0.9-5.8 months), and overall survival was 17.3 months (range, 1.9-21.3 months). The most common adverse events were neutropenia and anorexia. CONCLUSION: IP combination chemotherapy may be an acceptable option for patients with SBA. Further studies are warranted to determine the optimal chemotherapeutic regimen for SBA. PMID- 19132482 TI - Detection of micrometastases in peritoneal washings of gastric cancer patients by the reverse transcriptase polymerase chain reaction. AB - BACKGROUND: Gastric cancer patients with positive (+) peritoneal cytology have a prognosis similar to stage IV patients. We studied the ability of quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to detect peritoneal micrometastases in patients undergoing staging laparoscopy. METHODS: Peritoneal washings were obtained prospectively from 34 patients with gastric adenocarcinoma undergoing staging laparoscopy and 6 patients undergoing laparoscopy for benign disease. Each sample underwent cytologic and RT-PCR analysis for tumor markers: carcinoembryonic antigen (CEA), cytokeratin 20 (CK20), survivin, and MUC2. Markers were evaluated on the basis of their deviance from the ideal marker. RESULTS: Pathologic stages for the gastric cancer patients were: stage I, 9 (27%); stage II, 7 (21%); stage III, 15 (44%); and stage IV, 3 (9%). The four cytology (+) patients were: stage II, 1; stage III, 1; and stage IV, 2. Fifteen patients were RT-PCR (+), including all cytology (+) patients. The optimal threshold for cycle amplification was 35, based on a receiver operating characteristic curve. CEA had the smallest deviance. CONCLUSION: RT-PCR using a panel of tumor markers, including CEA, detects (+) cytology. The clinical significance of "false-positive" overexpression of CEA, survivin, or CK20 but cytology (-) remains to be defined. RT-PCR could represent a more sensitive method than cytology for detection of subclinical peritoneal tumor dissemination; this may be useful in improving patient selection for operative management and clinical trials. PMID- 19132483 TI - Recurrence in early gastric cancer with lymph node metastasis. AB - BACKGROUND: Early gastric cancer (EGC) has an excellent prognosis, but some patients with lymph node-positive disease will develop recurrence. In this study we investigated the risk factors for recurrence in this selected group of patients. METHODS: The clinical and pathological records of 2368 patients who underwent gastrectomy for solitary EGC between 1980 and 1999 at the National Cancer Center Hospital, Tokyo, were examined. Two hundred and thirty-eight patients (10%) were lymph node-positive (positive for lymph node metastasis) and form the population of this study. RESULTS: Nineteen (8%) of the 238 patients with lymph node-positive disease developed recurrence. The most common site of recurrence was lymph node (37%), followed by liver (21%). The interval between surgery and the detection of recurrence ranged from 3 to 98 months, with a median of 26 months. Multivariate analysis demonstrated that the number of metastatic nodes was an independent risk factor for recurrence. Patients with seven or more metastatic nodes had the highest rate of recurrence, at 38%. CONCLUSION: The number of nodes positive for metastasis was the only independent risk factor for recurrence after curative surgery in patients with lymph node-positive early gastric cancer. These high-risk patients may obtain additional survival benefit if targeted with adjuvant chemotherapy. PMID- 19132484 TI - Heterogeneous prognoses of patients with tumors invaded within muscularis propria according to tumor depth in the layers of the muscularis propria. AB - BACKGROUND: The muscularis propria of the stomach is histologically divided into three layers; namely, the innermost oblique, the inner circular, and the outer longitudinal layers. In patients with gastric cancer the depth of tumor invasion has been reported to correlate with lymph node metastasis and prognosis. However, it is unclear whether the depth of tumor invasion in the muscularis propria has an effect on lymph node metastasis and prognosis. METHODS: Fifty-nine gastric cancer patients with muscularis propria invasion were analyzed retrospectively. These patients were divided into two groups, the inner group, with invasion up to the inner circular layer; and the outer group, with invasion beyond the inner circular layer. The relationships between tumor invasion and clinicopathological factors and survival were evaluated. RESULTS: Of the 59 patients, 34 were classified as the inner group, and 25 were classified as the outer group. The inner group had a significantly lower probability of lymph node metastasis (P = 0.0053) and a significantly better overall cancer-specific survival (P = 0.017) than the outer group. CONCLUSION: Gastric cancers with muscularis propria invasion had heterogeneous prognoses according to the tumor depth in the muscularis propria layers. PMID- 19132485 TI - Initial experience in Brazil with endoscopic submucosal dissection for early gastric cancer using insulation-tipped knife: a safety and feasibility study. AB - BACKGROUND: Endoscopic resection is an adequate treatment for subgroups of patients with early gastric cancer. Endoscopic submucosal dissection (ESD) represents a recent advance and leads to good results when adequately indicated. There is great experience with this technique in Japan and it is gaining acceptance among Western endoscopists. We present the first gastric ESD series performed in Brazil. METHODS: Patients with well-differentiated adenocarcinomas macroscopically classified as early cancer, less than 30 mm with no ulcer or scar, were included. ESD was performed with an insulated-tip knife and electrosurgical unit with endocut mode. Clinicopathological aspects and morbidity were evaluated. The study was approved by the local ethics committee and informed consent was obtained from all participant subjects. RESULTS: From October 2005 to July 2007, 160 patients received surgical treatment for gastric cancer; 44 patients (27.5%) had early gastric cancer. In this latter group, 15 procedures (ESD) were performed in 12 patients. The mean size of the lesions was 16.8 mm. Almost half of the lesions were located in the proximal third of the stomach and the mean elapsed time for the procedure was 140 min. In 80% of the cases resection was en bloc and 80% of the resections were considered curative. We had three perforations, which were managed clinically, and no bleeding. CONCLUSION: When adequately indicated, ESD is a safe and feasible technique. PMID- 19132486 TI - Laparoscopy-assisted esophagoenteral anastomosis using endoscopic purse-string suture instrument "Endo-PSI (II)" and circular stapler. AB - It is said that laparoscopic esophagoenteral anastomosis is not easy. In particular, purse-string suture of the abdominal esophagus is difficult when using a circular stapler. We have developed an endoscopic purse-string suture instrument, the "Endo-PSI (II)", and the instrument was employed clinically during laparoscopy-assisted total gastrectomy. The device was inserted into the abdominal cavity through a 4-cm minilaparotomy of the epigastrium, and pneumoperitoneum was established by closing a Lap Disc. The Endo-PSI (II) was attached to the abdominal esophagus and a straight needle with a 2-0 polypropylene suture was passed through the device laparoscopically. After a purse-string suture of the abdominal esophagus was made, the abdominal esophagus was transected laparoscopically and the removed stomach was pulled out through the minilaparotomy. The anvil head of a circular stapler was inserted into the abdominal cavity through the minilaparotomy, and insertion of the anvil into the esophagus and ligation of the purse-string suture were performed laparoscopically, too. The combination of using a circular stapler for esophagojejunostomy and closure of the jejunal stump was also performed laparoscopically. Between May 2007 and May 2008, these products were used in 23 patients during laparoscopy-assisted total gastrectomy. There were no cases that required conversion to a conventional open procedure. The newly developed Endo PSI (II) was useful for laparoscopic purse-string suture of the esophagus. PMID- 19132487 TI - Intensity-modulated radiation therapy for orbital lymphoma. AB - PURPOSE: Orbital manifestations of non-Hodgkin's lymphoma (NHL) are rare and accounts for only 1% of all cases of NHL. There have been no reports of treating orbital lymphoma using intensity-modulated radiotherapy (IMRT). MATERIALS AND METHODS: Four patients were treated at our institution for orbital lymphoma using IMRT. Radiotherapy (RT) plans using wedged pair fields were developed for comparison. Clinical results using IMRT are presented and a dosimetric analysis between IMRT and RT was performed. RESULTS: All patients had a complete response based on their physical examinations and post-IMRT imaging. Symptoms that had been present at initial presentation resolved in all patients during the course of the treatment. All four patients experienced only grade 1 dry eye syndrome and keratitis. The average dose to the contralateral orbit, lacrimal gland, and lens were all significantly reduced (P < 0.01) in IMRT patients as compared to the RT patients. IMRT reduced the V5 and V10 for the contralateral lens, orbit, and lacrimal gland and the optic chiasm (P < 0.05). CONCLUSION: IMRT is feasible when treating orbital lymphoma and reduces dose to critical structures while providing excellent dose coverage of target volumes. IMRT offers patients with orbital lymphoma excellent clinical outcomes, similar to conventional RT, with no increased toxicity. PMID- 19132488 TI - Multidetector computed tomography diagnosis of primary and secondary epiploic appendagitis. AB - PURPOSE: The aim of this study was to evaluate the epiploic appendages in patients with acute abdomen using multidetector computed tomography (MDCT) and to determine the incidence of primary and secondary epiploic appendagitis (EA). MATERIALS AND METHODS: A radiologist reviewed MDCT images from 1338 patients with acute abdomen for visible epiploic appendages. Two radiologists then reviewed the MDCT images showing inflamed epiploic appendages and diagnosed primary EA, secondary EA, or other conditions by consensus. The CT criteria for primary EA are a round or oval pericolonic fatty lesion with a hyperattenuated rim and adjacent fat stranding, without other causes of inflammation. Secondary EA is diagnosed if an epiploic appendage is found to be due to inflammation from other inflammatory entities. RESULTS: Epiploic appendages were identified in 19 patients. Four patients (0.3%) had a retrospective CT diagnosis of primary EA. Twelve patients (0.9%) had a retrospective CT diagnosis of secondary EA (primary condition was diverticulitis in 10 patients and inflammatory bowel disease in 2 patients). The remaining three patients had calcification of an epiploic appendage suggestive of old EA. CONCLUSION: Primary EA should be included in the differential diagnosis of acute abdomen. Occasionally, inflammation of the epiploic appendages is secondary to other inflammatory conditions. PMID- 19132489 TI - Concurrent chemoradiotherapy with gemcitabine plus regional hyperthermia for locally advanced pancreatic carcinoma: initial experience. AB - PURPOSE: The aim of this study was to evaluate the efficacy and toxicity of concurrent chemoradiotherapy (CRT) with gemcitabine plus regional hyperthermia (HT) for locally advanced pancreatic carcinoma (LAPC). MATERIALS AND METHODS: A total of 29 patients with LAPC treated with concurrent CRT using gemcitabine were retrospectively analyzed. Radiotherapy was administered with a median total dose of 61.2 Gy. Of the 29 patients, 20 (69%) also underwent regional HT during CRT (CRHT group). The remaining 9 patients did not receive regional HT (CRT group) because of a common bile duct stent placement, patient refusal, older age, or obesity. The efficacy and toxicity of the treatments and the predictors of good outcome were evaluated. RESULTS: The median disease progression-free and overall survival times were significantly better for the CRHT group than for the CRT group (8.8 vs. 4.9 months, P = 0.02, and 18.6 vs. 9.6 months, P = 0.01), respectively. Grade 3-4 hematological toxicities for the CRHT group were detected in eight patients (40%) and grade 3 nonhematologic toxicity in one (diarrhea). CONCLUSION: Concurrent CRT using gemcitabine with regional HT may be a feasible and promising regimen for LAPC, and the results justified further evaluation in a large number of patients to confirm its definite benefit. PMID- 19132490 TI - Communication between cochlear perilymph and cerebrospinal fluid through the cochlear modiolus visualized after intratympanic administration of Gd-DTPA. AB - PURPOSE: Intratympanic injection of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) has been reported as a procedure to visualize endolymphatic hydrops of Meniere's disease. We frequently noted that cerebrospinal fluid (CSF) in the internal auditory canal (IAC) was also enhanced after this procedure. The purpose of this study was to evaluate how frequently this occurs and to investigate the specific features of patients who lack this communication. MATERIALS AND METHODS: A total of 25 patients with clinically suspected endolymphatic hydrops underwent the procedure. After 24 h, three-dimensional fluid-attenuated inversion recovery (3D-FLAIR) and 3D constructive interference in steady state (3D-CISS) were performed. The presence of contrast enhancement in the CSF space of the fundus of the IAC was evaluated. RESULTS: The contrast ratio between CSF of the IAC fundus and cerebellar white matter on the injected side was 1.49 +/- 0.65, and that of the noninjected side was 0.32 +/- 0.16 (P < 0.01). Enhancement of the CSF space in the IAC fundus was seen in all but two subjects: one had enlarged endolymphatic duct and sac syndrome (EEDS), and the other had cochlear nerve agenesis. In these two patients, the cochlear modiolus seemed to be normal. CONCLUSION: Intratympanic Gd-DTPA administration can reveal permeability of the modiolus and might facilitate evaluation of functional abnormalities of the modiolus not detected by conventional imaging tests. PMID- 19132491 TI - Percutaneous osteoplasty for hypervascular bone metastasis. AB - We report three cases of percutaneous osteoplasty for the treatment of hypervascular bone metastasis. Four hypervascular bone metastases were treated in three consecutive patients by percutaneous osteoplasty under fluoroscopic or computed tomographic guidance. Primary malignant tumors included pheochromocytoma and renal cell carcinoma. Pain relief after osteoplasty was achieved in all three patients. There was no major complication. Projectile bleeding and subcutaneous hematoma were noted during or after osteoplasty but were easily controlled conservatively. Percutaneous osteoplasty for hypervascular bone metastases is not only a highly effective but also a minimally invasive technique that provides immediate pain relief without major complication. PMID- 19132492 TI - Intraorbital lobular capillary hemangioma (pyogenic granuloma). AB - A 44-year-old man with a history of a foreign body in his right eye visited our hospital. On computed tomography a well-enhanced mass with low attenuation septal walls and a capsule was detected in the right orbit. On magnetic resonance imaging the mass showed inhomogenous high intensity on T2-weighted images and low intensity on diffusion-weighted images. The mass was histopathologically diagnosed as a lobular capillary hemangioma. This is the first report about image findings of lobular capillary hemangioma in the orbit. PMID- 19132493 TI - Computed tomographic and magnetic resonance features of inflammatory myofibroblastic tumor of the lung in children. AB - We report two cases of inflammatory myofibroblastic tumor (IMT) of the lung in a 4-year-old boy and a 7-year-old girl. We performed dynamic contrastenhancement computed tomography in both of our cases and dynamic contrast enhancement magnetic resonance imaging (MRI) in one case. These dynamic studies showed a demarcated mass with delayed enhancement in both cases. A T1-weighted MR image shows a mass with homogeneous low signal intensity, and a T2- weighted image shows a mass with slightly high signal intensity. A post-contrast-enhanced T1 weighted image demonstrates homogeneous enhancement. We speculate that the delayed enhancement could be attributed to the abundant fibrous tissue, which was the main structural material of the tumor. This same finding has been described in previous reports of IMT in other organs. PMID- 19132494 TI - Computed tomography of the gastrointestinal manifestation of hereditary angioedema. AB - We report a case of gastrointestinal manifestation of hereditary angioedema. Computed tomography (CT) revealed wall thickening of the gastric antrum, duodenum, and jejunum. Dilatation of the third part of the duodenum, thickening of the small bowel mesentery and omentum, and retroperitoneal edema were present. The importance of considering this condition in patients presenting such CT findings correlated with the appropriate history is discussed. PMID- 19132495 TI - Helical tomotherapy for total lymphoid irradiation. AB - Total lymphoid irradiation is employed in the preparative regimens for allogeneic bone marrow and solid organ transplantation, solid organ transplant rejection, and chronic graft-versus-host disease. Linear accelerator-based radiotherapy, typically involving opposed anteroposterior and posteroanterior beams, has been commonly used; however, extended source-to-skin patient setup and/or field matching are required, and all organs within the beam coverage receive the entire prescribed dose. Megavoltage helical tomotherapy represents a technological advance in terms of both treatment delivery and patient positioning. The continuously rotating multileaf collimated fan beam allows highly conformal coverage of complex target geometries, in turn allowing avoidance of radiosensitive adjacent organs. In addition, the megavoltage computed tomographic scans allow potentially more accurate, targetbased setup verification. The present case report describes tomotherapy-based total lymphoid irradiation in an adult patient with late-onset cardiac transplant rejection. Treatment planning allowed dose minimization to the spinal cord, kidneys, intestinal compartment, and lungs. The patient tolerated treatment well without acute adverse effects, and he is now in early follow-up. PMID- 19132498 TI - Highlights of a new type of intercellular communication: microvesicle-based information transfer. AB - Microvesicles (MVs) are membrane-covered cell fragments released by most cell types during apoptosis or activation. They are increasingly considered to play a pivotal role in information transfer between cells. Their presence and role have been proven in several physiological and pathological processes, such as immune modulation in inflammation and pregnancy, or blood coagulation and cancer. MVs represent a newly recognized system of intercellular communications. They not only may serve as prognostic markers in different diseases, but could also hold the potential to be new therapeutic targets or drug delivery systems. The present overview aims to highlight some aspects of this new means of cellular communication: "microvesicular communication". PMID- 19132496 TI - Ultra-low-dose computed tomography system with a flat panel detector: assessment of radiation dose reduction and spatial and low contrast resolution. AB - PURPOSE: The aim of this study was to introduce a prototype cone-beam computed tomography system equipped with a flat panel detector (FPD-CT system) and measure its radiation dose and spatial and lowcontrast resolution. MATERIALS AND METHODS: A patient was rotated in a sitting position, and cone beam data were acquired with the flat panel detector from a fixed X-ray tube. Absorbed dose, spatial and low-contrast resolution, and variation in the CT attenuation value were assessed quantitatively in the acrylic phantom. The visibility of normal blood vessels in clinical images of seven patients was analyzed qualitatively by five board certified radiologists. These quantitative and qualitative data were compared between the FPD-CT system and multidetector row CT (MDCT). RESULTS: Minimal low contrast sensitivity and a moderate spatial resolution were demonstrated in images of central lung fields acquired by FPD-CT. The absorbed dose in the FPD-CT system decreased to approximately 2.5% of the dose in the MDCT system. CONCLUSION: Considering crossover structures in normal blood vessels and bronchi in the central areas of lung fields, this result implies that fairly acceptable spatial resolution can be realized with FPD-CT for detection and frequent follow up of pulmonary abnormalities in the central lung fields. PMID- 19132499 TI - The effect of capillarisin on glycochenodeoxycholic acid-induced apoptosis and heme oxygenase-1 in rat primary hepatocytes. AB - The accumulation of hydrophobic bile acids plays a role in the induction of apoptosis and necrosis of hepatocytes during cholestasis. Glycochenodeoxycholate acid (GCDC) triggers a rapid oxidative stress response as an event of glutathione (GSH) depletion and nuclear factor kappa B (NF-kappaB) activation. We therefore investigated whether the bioactivity of the antioxidant capillarisin (Cap) prevents GCDC-induced hepatocyte damage. Isolated rat hepatocytes were co incubated with 100 muM GCDC and 0.5 mg/ml Cap for 4 h. GSH depletion and thiobarbituric acid-reactive substances (TBARS, measure of lipid peroxidation) increased after GCDC exposure, but were markedly suppressed by Cap treatment. Cap protected hepatocytes from a GCDC-induced increase in reactive oxygen species (ROS) generation and mitochondrial membrane potential induction, as measured by flow cytometry analysis. In addition, Cap was shown to inhibit GCDC-mediated NF kappaB activation by using electrophoretic mobility shift assays (EMSA). In contrast to GCDC, Cap not only significantly decreased cytochrome c release and caspase-3 enzyme activity, but also suppressed heme oxygenase-1 protein and mRNA expression in hepatocytes. These results demonstrate that Cap function as an antioxidant reduced hepatocyte injury caused by hydrophobic bile acids, perhaps by preventing generation of ROS and release of cytochrome c, thereby minimizing hepatocytes apoptosis. PMID- 19132500 TI - pH-sensitive nanoparticles self-assembled from a novel class of biodegradable amphiphilic copolymers based on chitosan. AB - A new type of biodegradable amphiphilic graft copolymers, PEG-g-hexanoyl chitosan, was synthesized by a facile scheme. The self-assemble properties of the copolymers were studied by TEM, fluorometry and dynamic light scattering. It was found that spherical nanoparticles of the copolymers could be formed through dialysis method. With the increase of PEG content in the copolymers, the average diameter of the nanoparticles decreased from about 180 to 40 nm. PEG fraction in the copolymers has little effect on the copolymer CAC. The micellization of the copolymers was strongly dependent of the medium pH. When pH was lowered from 7.2 to 6.8, the average diameter of the nanoparticles dramatically changed from about 180 to 60 nm. PMID- 19132501 TI - Strontium oxide doped quaternary glasses: effect on structure, degradation and cytocompatibility. AB - This preliminary study focuses on the effect of adding SrO to a Ti-containing quaternary phosphate glass system denoted by P(2)O(5)-Na(2)O-CaO-TiO(2). The following four different glass compositions were manufactured: 0.5P(2)O(5) 0.17Na(2)O-0.03TiO(2)-(0.3-x)CaO-xSrO where x = 0, 0.01, 0.03 and 0.05. Structural characterisation revealed glass transition temperatures in the range 427-437 degrees C and the presence of sodium calcium phosphate as the dominant phase in all the glasses. Degradation and ion release studies conducted over a 15 day period revealed that the Sr-containing glasses showed significantly higher degradation and ion release rates than the Sr-free glass. Cytocompatibility studies performed over a 7-day period using MG63 cells showed that the addition of 5 mol% SrO yielded glasses with cell viability nearly equivalent to that observed for quaternary TiO(2) glasses. PMID- 19132503 TI - Synthesis of high purity hydroxyapatite nanopowder via sol-gel combustion process. AB - A polymeric sol-gel combustion method has been used to synthesize nanocrystalline hydroxyapatite (HA) powder from calcium nitrate and triethyl phosphate with the addition of NH(4)OH. The sol-gel combustion process generates phase-pure nanocrystalline HA powder, as characterized using Fourier transform infrared (FTIR), X-ray diffraction (XRD), and transmission electron microscopy (TEM). Sintering of the HA powder compact at 1200 degrees C for 2 h leads to a 93% theoretical dense ceramic body. This method offers an easy route for the preparation of phase-pure nanocrystalline HA powder. PMID- 19132502 TI - Fibronectin modulates the morphology of osteoblast-like cells (MG-63) on nano grooved substrates. AB - Cell interactions with biomaterials are affected by surface topographic and chemical cues. Although it is well-known that nanometrical grooves/ridges structure modulates cellular spreading, elongation, and alignment, the combinational influence of surface topographic and chemical cues is not well studied. In this study, nano-textured silicon substrata with parallel ridges of 90, 250, or 500 nm wide, separated by grooves with equal width, were fabricated by electron beam lithography and dry etching techniques. Osteoblast-like cells, MG-63, were cultured on the patterned substrata with or without pre-adsorption of fibronectin. The cell morphology was imaged by scanning electron microscopy, and analyzed by image software. We found that FN coating initially modulated cellular spreading, length, and orientation on all types of grooved surfaces. However, after 24 h of culture, the cell morphology was not affected by FN coating on the 250-nm and 500-nm surfaces, while FN decreased cell alignment on the 90-nm surfaces. Our results suggest that surface chemical cues influence the initial cell-substratum contact, while the long-term cellular morphology is dictated by surface topographic cues. PMID- 19132504 TI - Biomimetic apatite formation on Ultra-High Molecular Weight Polyethylene (UHMWPE) using modified biomimetic solution. AB - Modifications were performed on a biomimetic solution (SBF), according to previous knowledge on the behavior of ions present in its composition, in order to obtain apatite coatings onto Ultra-High Molecular Weight Polyethylene (UHMWPE) without having to use polymer pre-treatments that could compromise its properties. UHMWPE substrates were immersed into a 30% H(2)O(2) solution for a 24 h period and then submitted to a biomimetic coating method using standard SBF and two other modified SBF solutions. Apatite coatings were only obtained onto UHMWPE when the modified SBF solutions were used. Based on these results, apatite coatings of biological importance (calcium-deficient hydroxyapatite-CDHA, amorphous calcium phosphate-ACP, octacalcium phosphate-OCP, and carbonated HA) can be obtained onto UHMWPE substrates, allowing an adequate conciliation between bonelike mechanical properties and bioactivity. PMID- 19132505 TI - Preparation and characterization of gelatin sponge millispheres from air-in-water in-oil-type emulsions. AB - A novel method for the preparation of gelatin sponge millispheres (GSMs) for biomaterials such as embolic agents and cell scaffolds was developed using an air in-water-in-oil-type emulsion. The droplets, consisting of a foamy gelatin suspension in caprylic triglyceride, were gelled and rinsed with isopropanol. Sonication and depressurization were used during the rinsing process to create interconnected pores. GSMs cross-links created over 4 h at 155 degrees C without any agent were insoluble and had short and long diameters of 1.1 +/- 0.2 mm and 1.3 +/- 0.2 mm, respectively. The residual isopropanol and caprylic triglyceride were <0.05% (w/w) and <1% (w/w) respectively. The level of bacterial endotoxins in the extracts was below 0.025 EU/ml, and no bacterial or fungal growth was found during sterility testing. The GSMs produced using this method were considered to meet the basic requirements of embolic agents. PMID- 19132506 TI - Biocompatibility study of theophylline/chitosan/beta-cyclodextrin microspheres as pulmonary delivery carriers. AB - To evaluate the biocompatibility of the theophylline/chitosan/beta-cyclodextrin microspheres, which has a potential application in pulmonary delivery system. The detection of LDH and protein in BALF was examined acute cell toxicity, hemolysis test was carried out to estimate blood toxicity; Micronucleus Test was reckoned to identify genotoxicity, MTT assay was used to evaluate in vitro cytotoxicity, and muscle implantation investigated the tissue biocompatibility. The results demonstrated that the total contents of protein and LDH in BALF were not significantly different from that of normal group. The experiments showed that the cytotoxicity was depended on the concentration and had no cytoxicity at low concentration and no hemolysis activity. The micronucleus frequency of MS B was 0.99 per thousand, which showed no genotoxic effects either. The results of implantation showed that the microspheres had no effect on hemoglobin and no toxicity in the liver and kidney. The inflammations of muscle tissue were not significantly different from that of operative suture, therefore, the MS B possess high good biocompatibility and can be applied in pulmonary sustained release systems. PMID- 19132507 TI - Study on compressive mechanical properties of nanohydroxyapatite reinforced poly(vinyl alcohol) gel composites as biomaterial. AB - Nanohydroxyapatite reinforced poly(vinyl alcohol) (nano-HA/PVA) gel composites has been proposed as a promising biomaterial to replace diseased or damaged articular cartilage. In this paper, nano-HA/PVA gel composites were prepared by in situ synthesis nano-HA particles in PVA solution and accompanied with freeze/thaw method. The influence of nano-HA content, PVA concentration and freeze/thaw cycle times on the compressive mechanical behavior of nano-HA/PVA gel composites were evaluated using mechanical test equipment. The results showed that the compressive mechanical behavior of nano-HA/PVA gel composites was similar to that of natural articular cartilage, which held special viscoelastic characteristics. Both the compressive strength and modulus of the composites improved correspondingly with the rise of freeze/thaw cycle times and PVA concentration. The compressive strength and modulus of nano-HA/PVA gel composites firstly increased and then presented decreasing trend with the rise of nano-HA content. Furthermore, the compressive modulus of the composites improved exponentially with the rise of compressive strain ratio. PMID- 19132508 TI - An in vitro release study of 5-fluoro-uracil (5-FU) from swellable poly-(2 hydroxyethyl methacrylate) (PHEMA) nanoparticles. AB - Nanomaterials are at the leading edge of the rapidly developing field of nanotechnology. The use of nanoparticles as drug delivery vehicles for anticancer therapeutics has great potential to revolutionize the future of cancer therapy. The present paper concerns both the optimizations of anticancer drug loading and its release from polymeric nanoparticles. The major aim of this study was to design poly (HEMA) nanoparticles as swelling controlled drug release system for anticancer drug. The prepared nanoparticles were characterized by Infra-Red (IR) Spectra, Particle size Analysis, and Scanning Electron Microscopy (SEM). The nanoparticles were loaded with widely used anticancer drug, 5-Fluorouracil, and controlled release of drug was investigated to observe the effects of various parameters such as percent loading of the drug, chemical architecture of the nanocarriers, pH, temperature, and nature of release media on the release profiles. The chemical stability of 5-Fluorouracil (5-FU) was also tested in phosphate buffer saline (PBS) (pH = 7.4) and release was studied in various simulated biological fluids. The prepared nanoparticles could provide a possible pathway for controlled and targeted delivery of anticancer drug, thus causing lower side effects and higher efficacy. PMID- 19132509 TI - Preparation of highly porous hydroxyapatite from cuttlefish bone. AB - Hydroxyapatite structures for tissue engineering applications have been produced by hydrothermal (HT) treatment of aragonite in the form of cuttlefish bone at 200 degrees C. Aragonite (CaCO(3)) monoliths were completely transformed into hydroxyapatite after 48 h of HT treatment. The substitution of CO(3) (2-) groups predominantly into the PO(4) (3-) sites of the Ca(10)(PO(4))(6)(OH)(2) structure was suggested by FT-IR spectroscopy and Rietveld structure refinement. The intensity of the nu(3)PO(4) (3-) bands increase, while the intensity of the nu(2)CO(3) (2-) bands decrease with the duration of HT treatment resulting in the formation of carbonate incorporating hydroxyapatite. The SEM micrographs have shown that the interconnected hollow structure with pillars connecting parallel lamellae in cuttlefish bone is maintained after conversion. Specific surface area (S (BET)) and total pore volume increased and mean pore size decreased by HT treatment. PMID- 19132510 TI - Feasibility of biodegradable PLGA common bile duct stents: an in vitro and in vivo study. AB - The current study investigates the feasibility of using a biodegradable polymeric stent in common bile duct (CBD) repair and reconstruction. Here, poly(L-lactide co-glycolide) (PLGA, molar ratio LA/GA = 80/20) was processed into a circular tube- and dumbbell-shaped specimens to determine the in vitro degradation behavior in bile. The morphology, weight loss, and molecular weight changes were then investigated in conjunction with evaluations of the mechanical properties of the specimen. Circular tube-shaped PLGA stents with X-ray opacity were subsequently used in common bile duct exploration (CBDE) and primary suturing in canine models. Next, X-ray images of CBD stents in vivo were compared and levels of serum liver enzymes and a histological analysis were conducted after stent transplantation. The results showed that the PLGA stents exhibited the required biomedical properties and spontaneously disappeared from CBDs in 4-5 weeks. The degradation period and function match the requirements in repair and reconstruction of CBDs to support the duct, guide bile drainage, and reduce T tube-related complications. PMID- 19132511 TI - Physicochemical characterisation of novel ultra-thin biodegradable scaffolds for peripheral nerve repair. AB - In this study, the physicochemical properties of microporous poly (epsilon caprolactone) (PCL) films and a composite material made of PCL and polylactic acid (PLA) blend were tested. Fabricated by solvent casting using dichloromethane, these ultra-thin films (60 +/- 5 microm in thickness) have a novel double-sided surface topography, i.e. a porous surface with pores 1-10 microm in diameter and a relatively smooth surface with nano-scaled texture. Porous surfaces were found to be associated with increased protein adsorption and the treatment of these polyester scaffolds with NaOH rendered them more hydrophilic. Differential Scanning Calorimetry (DSC) showed that the incorporation of PLA reduced the crystallinity of the original homopolymer. Chemical changes were investigated by means of Fourier Transform Infrared Spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). Average surface roughness (Ra), hydrophilicity/hydrophobicity and mechanical properties of these materials were also assessed for the suitability of these materials as nerve conduits. PMID- 19132512 TI - The preparation, cytocompatibility, and in vitro biodegradation study of pure beta-TCP on magnesium. AB - Biodegradable and bioactive beta-tricalcium phosphate (beta-TCP) coatings were prepared on magnesium (Mg) in order to improve its biocompatibility by a chemical method. The tensile bonding strength of beta-TCP coating and Mg substrate was measured by the standard adhesion test (ISO 13779-4). And the cytocompatibility of beta-TCP coated Mg was studied by using human osteoblast-like MG63 cells. It was found that the MG63 cells could grow well on the surface of beta-TCP coated Mg and the cell viability on beta-TCP coated Mg was above 80% during the cocultivation of MG63 cells and beta-TCP coated Mg for 10 days, indicating no cytotoxicity. It was concluded that the beta-TCP coated Mg had good cytocompatibility. The degradation of Mg substrate with beta-TCP coating in vitro was studied in detail by XRD, EDX, SEM, and ICP. The results showed that a bone like apatite continually formed on the surface of the sample with the degradation of both Mg substrate and beta-TCP coating in Hank's solution (a simulated body fluid). The biodegradation mechanism was preliminarily analyzed in the paper. PMID- 19132513 TI - Carboxymethyl derivative of scleroglucan: a novel thermosensitive hydrogel forming polysaccharide for drug delivery applications. AB - A carboxymethyl derivative of scleroglucan (Scl-CM) with a derivatization degree of 65 +/- 5% was synthesized. The rheological behaviour of this novel polymer was studied and compared with that of the starting polymer. We observed that the charged moieties carried on the chains could prevent the triple helix formation of Scl. Scl-CM aqueous solutions behave like true polymer solutions up to 1% w/v, whereas above this concentration a weak gel behaviour was observed. CaCl(2) addition to aqueous Scl-CM solutions led to a physical gel formation; the hydrogel strength was related to polymer and CaCl(2) concentrations. Temperature sweeps, registered at 1 Hz on hydrogels differing in CaCl(2) concentration, evidenced a gel --> sol transition in the range of 30-40 degrees C, depending on the molar ratio between carboxylic groups and Ca(+2). In order to verify a possible use of these hydrogels as drug delivery systems, acyclovir was loaded into the network. Rheological analysis evidenced that the loaded drug can affect the hydrogel elastic modulus. The release of acyclovir in phosphate buffer was evaluated at different temperatures in order to assess the suitability of this novel drug delivery system in topical applications. PMID- 19132514 TI - Novel platforms for oral drug delivery. AB - The aim of this review is to provide the reader general and inspiring prospects on recent and promising fields of innovation in oral drug delivery. Nowadays, inventive drug delivery systems vary from geometrically modified and modular matrices, more close to "classic" pharmaceutical manufacturing processes, to futuristic bio micro-electro-mechanical systems (bioMEMS), based on manufacturing techniques borrowed from electronics and other fields. In these technologies new materials and creative solutions are essential designing intelligent drug delivery systems able to release the required drug at the proper body location with the correct release rate. In particular, oral drug delivery systems of the future are expected to have a significant impact on the treatment of diseases, such as AIDS, cancer, malaria, diabetes requiring complex and multi-drug therapies, as well as on the life of patients, whose age and/or health status make necessary a multiple pharmacological approach. PMID- 19132515 TI - Combining MCMC with 'sequential' PKPD modelling. AB - We introduce a method for preventing unwanted feedback in Bayesian PKPD link models. We illustrate the approach using a simple example on a single individual, and subsequently demonstrate the ease with which it can be applied to more general settings. In particular, we look at the three 'sequential' population PKPD models examined by Zhang et al. (J Pharmacokinet Pharmacodyn 30:387-404, 2003; J Pharmacokinet Pharmacodyn 30:405-416, 2003), and provide graphical representations of these models to elucidate their structure. An important feature of our approach is that it allows uncertainty regarding the PK parameters to propagate through to inferences on the PD parameters. This is in contrast to standard two-stage approaches whereby 'plug-in' point estimates for either the population or the individual-specific PK parameters are required. PMID- 19132516 TI - Selective recognition of cobalt (II) ion by a new cryptand compound with N2O2S2 donor atom possessing 2-hydroxy-1-naphthylidene Schiff base moiety. AB - A new cryptand compound carrying 2-hydroxy-1-naphthylidene Schiff base moiety (3) was designed and synthesized by reaction of the corresponding macrobicyclic amine compound (1) and 2-hydroxy-1-naphthaldehyde (2). The influence of metal cations such as Mg2+, Ca2+, Sr2+, Fe2+,Co2+, Mn2+, Zn2+, Cd2+, Hg2+, Al3+ and Pb2+ on the spectroscopic properties of the new fluoroionophore was investigated in acetonitrile-dichloromethane solution (9.5/0.5) by means of absorption and emission spectrometry. The blue shifts on the fluorescence spectrum were observed for all metal cations at 504 nm. At the same time the fluorescence spectrum of the ligand showed quenching in the intensity of the signal at 504 nm for all metal cations except for Zn2+. Interaction of Co2+ with the ligand caused quenching of naphtyl fluorescence higher than 84%. The method showed good selectivity and sensitivity for Co2+ with respect to other metal cations with linear range and detection limit of 1.5 x 10(-7) to 3.3 x 10(-6) M and 4.8 x 10( 8) M respectively. PMID- 19132517 TI - Smoking cessation counseling practices of Texas pharmacists. AB - A cross-sectional study was conducted to determine pharmacists' awareness and education about smoking cessation and their communication with patients about smoking cessation. A survey was mailed to East Texas pharmacists practicing in the areas of hospital or clinical, retail or community, managed care, consultant, or academic pharmacy. Outcome measurements included: measures of the awareness of the 5 A's and 5 R's of smoking cessation, training received in smoking cessation, and communication practices regarding smoking cessation. There were 320 respondents. Approximately 10% of the respondents indicated they had received tobacco cessation counseling education during their formal educational training, 36% during continuing education programs, and 9% during both formal training and continuing education. About 44% reported they had received no tobacco cessation counseling training. Among pharmacists surveyed, 5% responded that they usually or always ask their patients if they smoke cigarettes, pipe, or cigars, 43% reported they sometimes or half of the time ask, and 45% said they never ask. There is a clear relationship between pharmacists awareness and education of smoking cessation techniques and their communication with patients about them. Pharmacy education leaders must continue their movement to include public health in the pharmacy curricula to produce pharmacists who are prepared to better serve the community. PMID- 19132518 TI - Richmond Latino needs assessment: a community-university partnership to identify health concerns and service needs for Latino youth. AB - The presence of Latinos in Virginia is a new phenomenon and as a result, less is known about the health needs of these newest community members. We formed a community-university partnership to identify health concerns and service needs as they relate to Latino youth living in Richmond, Virginia and the surrounding area. Using a mixed-method approach, survey data was obtained from 212 Latino adults, qualitative interviews were conducted with 15 community leaders and focus groups with 23 Latino parents (16 mothers, 7 fathers) and 6 Latino boys. Participants expressed concern about sexually transmitted infections/pregnancy among youth (76%), youth behavior problems at home/school (75%), and mental health problems (75%). Participants also expressed worry that youth would lose their connection to their Latin culture (83%). Qualitative data provided more information regarding these concerns by linking them with inter-ethnic tensions, and immigration and acculturation-related stressors. Survey participants also indicated a need for bilingual mental health services (88%) and after-school programs for youth (94%). This study provided the local community with information on the health concerns and service needs of a new group of community youth-Latino youth. Findings were presented to local community and City organizations that used the information to respond to the identified needs and/or concerns. The process in which the data was obtained may prove useful to other individuals interested in obtaining local level health information in emerging communities. PMID- 19132519 TI - Living at risk: the sibling's perspective of early-onset Alzheimer's disease. AB - Early-onset Alzheimer's disease (EOAD) is an increasingly diagnosed condition and is associated with genetic risk factors. This is one of the first studies exploring the lived experience of siblings of individuals with EOAD. We used structured questionnaires and semi-structured interviews to assess a broad range of siblings' experiences with and beliefs about EOAD, including knowledge, perceptions of personal risk, level of worry, and effects on life decisions. Participants (n = 24) were predominantly female (62.5%) and middle-aged (mean = 56.8 years; range 37-83). When asked about risk factors, genetics was cited most frequently (62.5%). Several potential means of reducing AD risk were endorsed, with 54% reporting engagement in behaviors for this purpose (e.g., keeping mentally active). Participants ranged widely in their perceived personal risk of AD (range: 0-100; mean = 35.6%), with higher perceived risk associated with worry about AD (p < 0.01). Understanding siblings' experiences with EOAD can inform how genetic counselors and healthcare professionals work with this population to facilitate risk communication and decision-making about testing and healthcare. PMID- 19132520 TI - Callous-unemotional traits and social information processing: multiple risk factor models for understanding aggressive behavior in antisocial youth. AB - This study examined multiple risk factor models of links among callous unemotional traits, aggression beliefs, social information processing, impulsivity, and aggressive behavior in a sample of 150 antisocial adolescents. Consistent with past research, results indicated that beliefs legitimizing aggression predicted social information processing biases and that social information processing biases mediated the effect of beliefs on aggressive behavior. Callous-unemotional traits accounted for unique variance in aggression above and beyond effects of more established risk factors of early onset of antisocial behavior, social information processing, and impulsivity. These findings add to recent research showing that callous-unemotional traits are a unique risk factor associated with aggression and criminal offending and suggest that targeting both affective and cognitive vulnerabilities may enhance clinical intervention with antisocial youth. PMID- 19132521 TI - CC16 inhibits the migration of eosinophils towards the formyl peptide fMLF but not towards PGD2. AB - Clara cell 16-kDa (CC16) is an anti-inflammatory protein chiefly produced in the lung epithelium. CC16 has been shown to inhibit the migration of rabbit neutrophils and human monocytes toward the formyl peptide N-formyl-methionine leucin-phenylalanin (fMLF). Eosinophils migrate towards prostaglandin D2 (PGD(2)) and CC16 has been shown to bind to PGD(2). Therefore we investigated if CC16 could inhibit the migration of human eosinophils and neutrophils towards fMLF and/or PGD(2). Migration of eosinophils and neutrophils was assessed in a microplate migration system using specific ligands and receptor antagonists. CC16 inhibited the migration of eosinophils and neutrophils toward fMLF, which is likely to result from the interaction of CC16 with members of the formyl-peptide receptor family. However, CC16 did not inhibit eosinophil migration towards PGD(2). We therefore propose that CC16 may down-modulate the entry of human eosinophils and neutrophils into the airways during inflammation in the lung. PMID- 19132522 TI - Psychiatric symptoms and psychosocial difficulties in young adults with autistic traits. AB - A screening version of the social responsiveness scale (SRS) was administered to 1,847 university students to identify a subgroup reporting significantly greater autism traits relative to their peers (High SRS group). A group reporting minimal autism traits was also identified (Low SRS group) matched for age, gender, and attentional difficulties. We administered the Behavioral Assessment System for Children-2nd edition (BASC-2), a comprehensive questionnaire designed to assess psychiatric symptoms and personality characteristics, to both groups. The high SRS group reported significantly more difficulties across the majority of areas, including depression/anxiety, interpersonal relationships, and personal adjustment. Thus, young adults reporting a greater degree of autistic traits also reported greater psychiatric difficulties across a wide psychosocial range. PMID- 19132523 TI - Assessment of physical self-concept in adolescents with intellectual disability: content and factor validity of the very Short Form of the Physical Self Inventory. AB - The purpose of this study was to test the factor validity and reliability of the Very Short Form of the Physical Self-Inventory- (PSI-VSF) within a sample of adolescents with mild to moderate Intellectual Disability (ID). A total of 362 ID adolescents were involved in two studies. In Study 1, the content and format scale response of the PSI-VSF were adapted for adolescents with ID. This instrument was thus renamed PSI-VSF-ID and two versions with two alternative responses scales format, were developed: Likert and graphical. In Study 2, results provided support for: (1) the factorial validity and reliability; and (2) factorial invariance across gender, age, type of school placement and ID level of the PSI-VSF-ID associated with a graphical response scale format. PMID- 19132524 TI - Molecular mutations associated with the in vitro passage of virulent porcine reproductive and respiratory syndrome virus. AB - Mutants of a highly pathogenic, porcine reproductive, and respiratory syndrome virus (PRRSV), JXA1 strain, were prepared by continuous in vitro passage. Genomic sequence comparisons were made between mutants obtained at different passages and the parental strain JXA1. The mutant strain obtained at passage 80 contained a 12 nucleotide insertion and 108 nucleotide mutations that resulted in 45 amino acid changes. Most of these changes (89%) occurred between passage 10 and 45 and were genetically stable for the next 35-70 passages. A comparison of the mutants, their parental strain, and several American PRRSV strains, identified 13 characteristic amino acid changes. These sites, as well as the distinct 12 nucleotide insertion, represent possible genetic markers for the evaluation of live vaccine applications, particularly for additional studies of the safety and potency of live PRRSV vaccines. PMID- 19132526 TI - A tryptophan hydroxylase 2 gene polymorphism is associated with panic disorder. AB - Panic disorder (PD) is a complex and heterogeneous psychiatric condition. Dysfunction within the serotonergic system has been hypothesized to play an important role in PD. The novel brain-specific serotonin synthesizing enzyme, tryptophan hydroxylase 2 (TPH2), which represents the rate-limiting enzyme of serotonin production in the brain, may therefore be of particular importance in PD. We investigated the TPH2 703G/T SNP for association with PD. Patients with PD (n = 108), and control subjects (n = 247), were genotyped for rs4570625 (TPH2 703G/T). Male and female subjects were analyzed separately. The severity of their symptoms was measured using the Spielberger state-trait anxiety inventory (STAI), panic disorder severity scale (PDSS), anxiety sensitivity index (ASI), acute panic inventory (API), and Hamilton's rating scale for depression (HAMD). The genotype and allele frequencies of the PD patients and controls were analyzed using chi(2) statistics. There was a significant difference in the allele frequency in rs4570625 between the PD patients and normal controls. The T allele was significantly less frequent in the PD patients. We also found a significant association with rs4570625 in the female subgroup. There was no difference in symptom severity among the genotypes of this polymorphism. This result suggests that rs4570625 polymorphism may play a significant role in the pathogenesis of PD. Moreover, rs4570625 may have a gender-dependent effect on susceptibility to PD. Further studies are needed to replicate the association that we observed. PMID- 19132527 TI - Adrenocortical functioning in boys with attention-deficit/hyperactivity disorder: examining subtypes of ADHD and associated comorbid conditions. AB - Disruptions to hypothalamic-pituitary-adrenal (HPA) axis function have been associated with varying forms of psychopathology in children. Studies suggesting children with ADHD have blunted HPA function have been complicated by the prevalence of comorbid diagnoses and heterogeneity of ADHD. The goals of this research were to assess the relations between waking and stress-response salivary cortisol levels and comorbid disruptive behavior (DBD) and anxiety (AnxD) disorders and problems in boys with ADHD, and to examine whether cortisol levels varied across ADHD subtypes. One hundred seventy elementary school-age boys with ADHD provided salivary cortisol at waking and in reaction to venipuncture. Parent reports were used to assess boys' psychiatric diagnoses and severity of behavioral problems. Boys' comorbid AnxD and anxiety problems were associated with greater cortisol reactivity, whereas boys' comorbid DBD and oppositional problems predicted diminished adrenocortical activity. Reactive cortisol increases were greatest in boys with ADHD and comorbid AnxD, but without DBD. ADHD subtypes were not differentially associated with waking, pre-stress baseline, or reactive cortisol levels. However, comorbid DBD predicted decreased cortisol reactivity in boys with inattentive and hyperactive subtypes of ADHD, but not in boys with combined subtype of ADHD. The results clarify previous patterns of distinct and divergent dysregulations of HPA function associated with boys' varying kinds of psychopathology. PMID- 19132529 TI - Opposite effects of protein kinase C beta1 (PKCbeta1) and PKCepsilon in the metastatic potential of a breast cancer murine model. AB - In this paper we investigated whether protein kinase C (PKC) beta1 and PKCepsilon, members of the classical and novel PKC family, respectively, induce phenotypic alterations that could be associated with tumor progression and metastatic dissemination in a murine model of breast cancer. Stable overexpression of PKCbeta1 in LM3 cells altered their ability to proliferate, adhere, and survive, and impaired their tumorigenicity and metastatic capacity. Moreover, PKCbeta1 induced the re-expression of fibronectin, an extracellular matrix glycoprotein which loss has been associated with the acquisition of a transformed phenotype in different cell models, and exerted an important inhibition on proteases production, effects that probably impact on LM3 invasiveness and dissemination. Conversely, PKCepsilon overexpression enhanced LM3 survival, anchorage-independent growth, and caused a significant increase in spontaneous lung metastasis. Our results suggest PKCbeta1 functions as an inhibitory protein for tumor growth and metastasis dissemination whereas PKCepsilon drives metastatic dissemination without affecting primary tumor growth. PMID- 19132528 TI - Enhanced dopamine D1 and D2 receptor gene expression in the hippocampus of hypoglycaemic and diabetic rats. AB - Hypoglycaemic coma and brain injury are potential complications of insulin therapy. Hippocampal neurons are particularly vulnerable to hypoglycaemic stress leading to memory impairment. In the present article, we have investigated the dopamine (DA) content, homovanillic acid (HVA)/DA turnover ratio, DA D(1) and DA D(2) receptors in the hippocampus of insulin-induced hypoglycaemic (IIH) and streptozotocin induced diabetic rats where brain functions are impaired. The DA content decreased significantly in hippocampus of diabetic, diabetic +IIH and control +IIH rats compared to control. The HVA/DA turnover ratio also increased significantly in diabetic, diabetic +IIH and control +IIH rats compared to control. Scatchard analysis using [(3)H] DA in the hippocampus showed a significant increase in DA receptors of diabetic, diabetic +IIH and control +IIH rats with decreased affinity. Gene expression studies using Real-time PCR showed an increased expression of DA D(1) and DA D(2) receptors in the hippocampus of hypoglycaemic and diabetic rats. Our results indicate that the dopaminergic system is impaired in the hippocampus of hypoglycaemic and hyperglycaemic rats impairing DA related functions of hippocampus. We observed a prominent dopaminergic functional disturbance in the hypoglycaemic condition than in hyperglycaemia compared to control. This dopaminergic dysfunction in hippocampus during hypoglycaemia and hyperglycaemia is suggested to contribute to cognitive and memory deficits. This will have clinical significance in the treatment of diabetes. PMID- 19132530 TI - Sepsis associated encephalopathy studied by MRI and cerebral spinal fluid S100B measurement. AB - The pathogenesis of sepsis associated encephalopathy (SAE) is not yet clear: the blood-brain barrier (BBB) disruption has been indicated among the possible causative mechanisms. S100B, a calcium binding protein, originates in the central nervous system but it can be also produced by extra-cerebral sources; it is passively released from damaged glial cells and neurons; it has limited passage through the BBB. We aimed to demonstrate BBB damage as part of the pathogenesis of SAE by cerebral spinal fluid (CSF) and serum S100B measurements and by magnetic resonance imaging (MRI). This paper describes four septic patients in whom SAE was clinically evident, who underwent MRI and S100B measurement. We have not found any evidence of CSF-S100B increase. Serum S100B increase was found in three out of four patients. MRI did not identify images attributable to BBB disruption but vasogenic edema, probably caused by an alteration of autoregulation, was diagnosed. S100B does not increase in CSF of septic patients; S100B increase in serum may be due to extracerebral sources and does not prove any injury of BBB. MRI can exclude other cerebral pathologies causing brain dysfunction but is not specific of SAE. BBB damage may be numbered among the contributors of SAE, which aetiology is certainly multifactorial: an interplay between the toxic mediators involved in sepsis and the indirect effects of hyperthermia, hypossia and hypoperfusion. PMID- 19132532 TI - Increased expression of beta-catenin in brain microvessels of a segmentally trisomic (Ts65Dn) mouse model of Down syndrome. AB - We examined the distribution of beta-catenin and endogenous blood serum albumin at the ultrastructural level in blood microvessels (capillaries) from brains of control and trisomic Ts65Dn mice. Morphological examination revealed an increased immunolabeling for beta-catenin in endothelial substructures of the capillary network, such as intercellular junctions, cytoplasm, and nuclei. These immunosignals were significantly increased in all endothelial substructures from trisomic mice. These changes, however, did not affect the blood-brain barrier function of the entire microvascular network, because the increased uptake of albumin by endothelial cells and the eventual escape of this protein (microleakage) into the perivascular neuropil were noted only in a few capillary profiles. Nevertheless, these findings suggest the involvement of some segments of the microvascular network in the brain pathology associated with DS. PMID- 19132533 TI - O-GlcNAc modification of radial glial vimentin filaments in the developing chick brain. AB - We examined the post-translational modification of intracellular proteins by beta O-linked N-acetylglucosamine (O-GlcNAc) with regard to neurofilament phosphorylation in the developing chick optic tectum. A regulated developmental pattern of O-GlcNAcylation was discovered in the developing brain. Most notably, discernible staining occurs along radial glial filaments but not along neuronal filaments in vivo. Immunohistochemical analyses in sections of progressive stages of development suggest upregulation of O-GlcNAc in the ependyma, tectofugal neuron bodies, and radial glial processes, but not in axons. In contrast, double label immunostaining of monolayer cultures made from dissociated embryonic day (E) 7 optic tecta revealed O-GlcNAcylation of most axons. Labeling of brain sections together with Western blot analyses showed O-GlcNAc modification of a few discrete proteins throughout development, and suggested vimentin as the protein in radial glia. Immunoprecipitation of vimentin from E9 whole brain lysates confirmed O-GlcNAcylation of vimentin in development. These results indicate a regulated pattern of O-GlcNAc modification of vimentin filaments, which in turn suggests a role for O-GlcNAc-modified intermediate filaments in radial glia, but not in neurons during brain development. The control mechanisms that regulate this pattern in vivo, however, are disrupted when cells are placed in vitro. PMID- 19132534 TI - Direct interaction of SNARE complex binding protein synaphin/complexin with calcium sensor synaptotagmin 1. AB - Although the binding of synaphin (also called complexin) to the soluble N ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex is critical for synaptic vesicle exocytosis, the exact role of synaphin remains unclear. Here, we show that synaphin directly binds to synaptotagmin 1, a major Ca(2+) sensor for fast neurotransmitter release, in a 1:1 stoichiometry. Mapping of the synaphin site involved in synaptotagmin 1 binding revealed that the C terminal region is essential for the interaction between these two proteins. Binding was sensitive to ionic strength, suggesting the involvement of charged residues in the C-terminus region. Mutation of the seven consecutive glutamic acid residues (residues 108-114) at the C-terminal region of synaphin to alanines or glutamines resulted in a dramatic reduction in synaptotagmin 1 binding activity. Furthermore, a peptide from the C-terminus of synaphin (residues 91 124) blocked the binding of synaptotagmin 1 to synaphin, an effect that was abolished by mutating the consecutive glutamic acid residues to alanine. Immunoprecipitation experiments with brain membrane extracts showed the presence of a complex consisting of synaphin, synaptotagmin 1, and SNAREs. We propose that synaphin recruits synaptotagmin 1 to the SNARE-based fusion complex and synergistically functions with synaptotagmin 1 in mediating fast synaptic vesicle exocytosis. PMID- 19132535 TI - Persons and their parts: new reproductive technologies and risks of commodification. AB - This paper explores one aspect of the social implications of new reproductive technologies, namely, the impact such technologies have on our understandings of family structures and our expectations of children. In particular it considers whether the possibilities afforded by such technologies result in a more contractual and commodified understanding of children. To do this the paper outlines the possibilities afforded by NRTs and their commodificatory tendencies; second, it explores the commodification debate using the somewhat parallel example of commodification of organs; and third, in light of these debates the link between the commodification of body parts and persons is addressed. It will argue that there is a prime facie connection between body parts and persons and thus, although needing to be balanced with other ethically relevant factors, commodification remains an issue of ethical concern. Accordingly we should only be supporting potentially commodifying practices when there are ethically pressing reasons to do so (such as in organ transplantation). Moreover given this link between body part and persons we should attempt to lessen commodifying attitudes and thus should resist the increasing use of practices which regard children as having choose-able parts. PMID- 19132536 TI - Change in criteria for USP dissolution performance verification tests. AB - The US Pharmacopeial Convention has been evaluating its performance verification tests (PVT) for several years. These tests help ensure the integrity of the US Pharmacopeia performance test when a dissolution procedure, as described in General Chapter Dissolution <711>, is relied upon to test a nonsolution orally administered dosage form. One result of the evaluation is a change in the PVT criterion from one based on individual tablet results to one based on the mean and variability of a set of tablets. This paper describes the new PVT and its criterion and how its acceptance limits are derived from results of a collaborative study, explains a two-stage option for the test, and presents operating characteristics. PMID- 19132537 TI - Polymeric matrix system for prolonged delivery of tramadol hydrochloride, part I: physicochemical evaluation. AB - Management of moderate or severe chronic pain conditions is the burden of clinicians dealing with patients trying to improve their quality of life and diminish their suffering. Although not a new opioid, tramadol has been recently rediscovered and widely used; this may be due to its favorable chronic safety and dependence profiles together with its high potency. Tramadol is a centrally acting analgesic with half-life of approximately 6 h; therefore, it requires frequent dosing. It is freely soluble in water; hence, judicious selection of retarding formulations is necessary. The current study is focused on the innovation of a novel, simple, monolayer, easy-to-use, cost-effective, and aesthetically acceptable bioadhesive transdermal delivery system overcoming the defects of the conventional "patch" as carrier system for tramadol, ensuring its adequate delivery, along with the physicochemical evaluation of the designed formulations. Monolithic tramadol matrix films of chitosan, different types of Eudragit, and binary mixtures of both were prepared. As a single-polymer film, chitosan film showed best properties except for somewhat high moisture uptake capacity, insufficient strength and rapid release, and permeation. Polymer blends were monitored in order to optimize both properties and performance. Promising results were obtained, with chitosan-Eudragit NE30D (1:1) film showing the most desirable combined, sufficiently rapid as well as prolonged release and permeation profiles along with satisfactory organoleptic and physicochemical properties. PMID- 19132540 TI - Assessing suturing skills in a self-guided learning setting: absolute symmetry error. AB - Directed self-guidance, whereby trainees independently practice a skill-set in a structured setting, may be an effective technique for novice training. Currently, however, most evaluation methods require an expert to be present during practice. The study aim was to determine if absolute symmetry error, a clinically important measure that can be assessed by the trainee, is a feasible assessment tool for self-guided learning of suturing skill. Forty-eight undergraduate medical trainees independently practiced suturing and knot tying skills using a benchtop model. Performance on a pretest, posttest, retention test and a transfer test was assessed using (1) the validated final product analysis (FPA), (2) the surgical efficiency score (SES), a combination of the FPA and hand motion analysis and (3) absolute symmetry error, a new measure that assesses the symmetry of the final product. Absolute symmetry error, along with the other objective assessment tools, detected improvements in performance from pretest to posttest (P < 0.05). A battery of correlation analyses indicated that absolute symmetry error correlates moderately with the FPA and SES. The development of valid, reliable and feasible technical skill assessments is needed to ensure all training centers evaluate trainee performance in a standardized fashion. Measures that do not require the use of experts or computers have potential for widespread use. We suggest that absolute symmetry error is a useful approximation of novices' suturing and knot tying performance. Future research should evaluate whether absolute symmetry error can enhance learning when used as a source of feedback during self-guided practice. PMID- 19132538 TI - Targeted delivery of nucleic-acid-based therapeutics to the pulmonary circulation. AB - Targeted delivery of functional nucleic acids (genes and oligonucleotides) to pulmonary endothelium may become a novel therapy for the treatment of various types of lung diseases. It may also provide a new research tool to study the functions and regulation of novel genes in pulmonary endothelium. Its success is largely dependent on the development of a vehicle that is capable of efficient pulmonary delivery with minimal toxicity. This review summarizes the recent progress that has been made in our laboratory along these research directions. Factors that affect pulmonary nucleic acids delivery are also discussed. PMID- 19132539 TI - Maximizing resources to study an uncommon cancer: E2C2--Epidemiology of Endometrial Cancer Consortium. AB - Endometrial cancer affects more than 40,000 women a year in the U.S. While the association of this disease with high body mass index and sex steroid hormones is well known, there are many questions about etiology that have not been resolved. Little is known about the genetic basis for risk associated with hormones or obesity, other common genetic factors associated with risk, or gene-environment interactions. E2C2, the Epidemiology of Endometrial Cancer Consortium, was formed in 2006 to provide a collaborative environment for addressing these questions by pooling data from existing studies. This allows for investigations of uncommon risk factors, risk for rare histologic subtypes, and associations within strata that cannot be achieved in individual studies. This report describes the establishment of the consortium, ongoing projects that demonstrate the advantages of collaborative efforts, and challenges faced. Overall, the consortium promises to provide an important means of furthering our knowledge about this cancer. PMID- 19132541 TI - The ABC-transporter AtmA is involved in nickel and cobalt resistance of Cupriavidus metallidurans strain CH34. AB - Cupriavidus metallidurans CH34 genome contains an ortholog of Atm1p named AtmA (Rmet_0391, YP_582546). In Saccharomyces cerevisiae, the ABC-type transport system Atm1p is involved in export of iron-sulfur clusters from mitochondria into the cytoplasm for assembly of cytoplasmic iron-sulfur containing proteins. An atmA mutant of C. metallidurans was sensitive to nickel and cobalt but not iron cations. AtmA increased also resistance to these cations in Escherichia coli strains that carry deletions of the genes for other nickel and cobalt transport systems. In C. metallidurans, atmA expression was not significantly induced by nickel and cobalt, but repressed by zinc. AtmA was purified as a 70 kDa protein after expression in E. coli. ATPase activity of AtmA was stimulated by nickel and cobalt. PMID- 19132542 TI - Assessing job satisfaction and stress among pharmacists in Northern Ireland. AB - OBJECTIVE AND SETTING: The pharmacy profession in Northern Ireland (NI) is currently experiencing major changes in practice development. The aim of this study was to determine the levels of job satisfaction and stress in pharmacists in NI in the context of these changes. METHOD: A questionnaire was adapted from one previously used in the United States (US). Following minor amendments and piloting, the survey was distributed by mail on two occasions (January/February 2007) to all registered community and hospital pharmacists in NI (n = 1,965). Data were entered into SPSS (version 15) and analysed using descriptive statistics, t-test and regression analysis. The significance level was set at P < 0.05. RESULTS: The overall response rate was 39% (n = 766). Of the potentially highest stress score of 165, overall mean scores were significantly higher for community than hospital pharmacists (P < 0.05). Both groups found interruptions, excessive workload, and inadequate staffing to be the most stressful aspects of their employment. Just over 30% (n = 178) of community pharmacists and half of all hospital pharmacists cited feeling often or frequently stressed because of imminent changes in contractual or organisational arrangements. Regression analysis indicated that sector of work (community or hospital) appeared to account for differences in self-reported stress. CONCLUSION: Any developments in professional practice need to be considered in the context of the well-being of the health professionals who implement, and are affected by, the changes. Unless stress in pharmacy is recognised and reduced, pharmacists and patients may be at risk. PMID- 19132543 TI - Incidental extracardiac findings at cardiac CT angiography: comparison of prevalence and clinical significance between precontrast low-dose whole thoracic scan and postcontrast retrospective ECG-gated cardiac scan. AB - PURPOSE: To compare the prevalence and clinical significance of incidental extracardiac findings at cardiac CT angiography (CCTA) with precontrast low-dose whole thoracic scan (LDCT) and ECG-gated CCTA. MATERIALS AND METHODS: We reviewed 254 patients who underwent CCTA. All participants first underwent LDCT to determine a range for CCTA and to screen unrecognized extracardiac lesions. CCTA was reconstructed with a small field of view of the heart. Clinically significant extracardiac findings were defined as abnormalities requiring further diagnostic work up, therapeutic intervention, or follow-up. RESULTS: On LDCT, 285 extracardiac findings were detected in 62.6% patients; on CCTA, 18 findings in 7% patients. Among these, 66 findings in 20.4% patients were considered clinically significant on LDCT, and 4 findings in 1.6% patients on CCTA. CONCLUSION: Clinically significant extracardiac findings are common in patients undergoing CCTA with a considerable number of extracardiac findings being detected only on LDCT. We advise performing whole thorax LDCT prior to CCTA. PMID- 19132544 TI - Suggestion for a new image-based aortic wall stiffness evaluation technique: arterial wall stiffness index. AB - This study evaluated arterial wall stiffness independent of variant background blood pressure. A new technique-arterial wall stiffness index (AWSI)-was developed and its use verified. Intraluminal pressure and luminal volume were measured on eight swine descending aortas. AWSI was formulated to evaluate absolute arterial wall stiffness independent of variable blood pressure and aortic size. AWSI variability with pressure change was compared with other wall stiffness evaluation parameters. AWSI determined from 100 descending aortic cine CT images and 108 carotid artery ultrasonography datasets were compared with age and Framingham risk score, respectively. Between 50 and 360 mmHg blood pressures, AWSI variance was 5.43% compared to 64.99% for classical compliance. AWSI correlated better with Framingham risk score and age than conservative wall stiffness evaluation methods. AWSI is a suitable method to evaluate arterial wall properties independent of variable background blood pressure and aortic size effects. PMID- 19132545 TI - Left ventricular myocardial volumes measured during arterial and delayed phases of multidetector row computed tomography: a study on intra- and interobserver variability. AB - To compare and correlate left ventricular (LV) myocardial volumes obtained using arterial and delayed phases of multidetector row computed tomography (CT) and evaluate their intra- and interobserver variation. Two observers evaluated the arterial- and delayed-phase serial short-axis images of 45 healthy volunteers. Intra- and interobserver variations in LV myocardial volumes were correlated with four factors-myocardial volume, contrast-volume-to-body-weight ratio, and contrast-to-noise ratios in the arterial and delayed phases. Variations in the apex, mid-ventricle, and base were compared. Intra- and interobserver analyzes revealed no statistical difference and good correlation. Intra- and interobserver variations were within 5 and 10%, respectively, and were independent of the four factors. Variations were the highest at the apex. LV myocardial volumes measured using arterial- and delayed-phase cardiac CT exhibit no significant difference and good correlation. Intra- and interobserver variations are both clinically acceptable, and the apex contributes most to these variations. PMID- 19132546 TI - Myocardial perfusion cardiac magnetic resonance for the diagnosis of coronary artery disease: do we need rest images? AB - Adenosine stress cardiovascular magnetic resonance (CMR) has been reported to be useful for the diagnosis of coronary artery disease (CAD). Most studies use rest and stress perfusion images. The objectives of this study were to determine (Barkhausen et al. in J Magn Reson Imaging 19(6):750-757, 1) the accuracy of visual assessment and myocardial perfusion reserve index (MPRI) in the diagnosis of CAD and (Rieber et al. in Fur Heart J 27(12):1465-1471, 2) the accuracy of analysis based on rest-stress and stress images. We enrolled patients with suspected CAD and referred them for coronary angiography (CAG). All the patients underwent adenosine stress CMR before CAG. Rest and stress perfusion images were analyzed by calculation of MPRI and visual assessment separately. Visual assessment was performed separately by using rest and stress images and by using only stress images. CAG was considered the gold standard. Sensitivity, specificity, and accuracy of both methods were calculated and compared. A total of 66 patients (mean age, 61.3 +/- 11.7 years) were studied. Thirty-eight patients (57.6%) were diagnosed with CAD. The sensitivity and specificity for the diagnosis of CAD (> or =50% stenosis) were 89.5 and 78.6% for MPRI, 76.3 and 75% for stress-rest visual method, and 86.8 and 75% for stress visual method, respectively. CMR perfusion had a relatively lower accuracy in patients with left ventricular systolic dysfunction, high left ventricular mass, or presence of late gadolinium enhancement than in patients without those CMR findings. Visual assessment of stress image of CMR perfusion is accurate and comparable to MPRI for the detection of CAD. PMID- 19132547 TI - In vivo magnetic resonance imaging of injected mesenchymal stem cells in rat myocardial infarction; simultaneous cell tracking and left ventricular function measurement. AB - To determine whether magnetic resonance imaging (MRI) can enable magnetically labeled mesenchymal stem cell (MSC) tracking and simultaneous in vivo functional data acquisition in rat models of myocardial infarction. Superparamagnetic iron oxide-laden human MSCs were injected into rat myocardium infarcted by cryoinjury 3 weeks after myocardial infarction. The control group received cell-free media injection. Before injection and for 3 months after, in vivo serial MRI was performed. Electrocardiography-gated gradient echo sequence MRI and cine MRI were performed for in vivo cell tracking and assessing cardiac function using left ventricular ejection fraction (LVEF), respectively. MRI revealed a persistent signal-void representing iron-laden MSCs until ten post-injection weeks. Serial follow-up MRI revealed that LVEF was significantly higher in the MSC injection group than in the control group. We conclude that MRI enables in vivo tracking of injected cells and evaluation of the long-term therapeutic potential of MSCs for myocardial infarction. PMID- 19132548 TI - Should we select for genetic moral enhancement? A thought experiment using the MoralKinder (MK+) haplotype. AB - By using preimplantation haplotype diagnosis, prospective parents are able to select embryos to implant through in vitro fertilization. If we knew that the naturally-occurring (but theoretical) MoralKinder (MK+) haplotype would predispose individuals to a higher level of morality than average, is it permissible or obligatory to select for the MK+ haplotype? I.e., is it moral to select for morality? This paper explores the various potential issues that could arise from genetic moral enhancement. PMID- 19132549 TI - Palliative sedation until death: an approach from Kant's ethics of virtue. AB - This paper is concerned with the moral justification for palliative sedation until death. Palliative sedation involves the intentional lowering of consciousness for the relief of untreatable symptoms. The paper focuses on the moral problems surrounding the intentional lowering of consciousness until death itself, rather than possible adjacent life-shortening effects. Starting from a Kantian perspective on virtue, it is shown that continuous deep sedation until death (CDS) does not conflict with the perfect duty of moral self-preservation because CDS does not destroy capacities for agency. In addition, it is argued that CDS can frustrate the imperfect duty of self-cultivation by reducing consciousness permanently. Nevertheless, there are cases where CDS is morally acceptable, namely, cases where the agent has already permanently lost the possibility for free action in advance of sedation--for example, due to excruciating and ongoing pain. Because the latter can be difficult to diagnose properly, safeguards may be needed in order to prevent the application of CDS for the wrong reasons. PMID- 19132551 TI - Fresh-frozen human bone graft for repair of defect after adenomatoid odontogenic tumour removal. AB - The aim of this paper was report the clinical, radiographic, and histological case of adenomatoid odontogenic tumour (AOT) in adolescent woman as well as present the reconstructive treatment of AOT using fresh-frozen human bone graft with guided bone regeneration. AOT is a benign, noninvasive lesion with slow but progressive growth. Biopsy and microscopic examination confirmed the presence of an AOT. Treatment was conservative and the prognosis was excellent. The patient has been followed-up for without recurrence. The use of fresh-frozen human bone graft can be a safe choice for reconstruction of the bone defects to treat AOT. PMID- 19132552 TI - The expense for one implantation of a banked bone allograft from a cadaveric donor and the issues affecting current advanced medical treatment in the Japanese orthopaedic field. AB - Demand for banked bone allografts is increasing in Japan; however, there are too few bone banks and the bone bank network is not well-established. One reason for this was lack of funding for banks. Bone banks had to bear all material expenses of banked bone allografts themselves because this was not designated a covered expense. In December 2004, the Japanese government started a new "Advanced Medical Treatment" administration system which allowed an approved institution to charge the expense of authorized advanced medical treatments directly to patients. The treatment named "Cryopreserved allogenic bone and ligamentous tissue retrieved from cadaveric donor" was approved as an advanced medical treatment in March 2007. We present the calculation method and the expense per implantation of a banked bone allograft from a cadaveric donor under this treatment and raise issues which affect this advanced medical treatment and remain to be resolved in the Japanese orthopaedic field. PMID- 19132550 TI - Exploring household income as a predictor of psychological well-being among long term colorectal cancer survivors. AB - PURPOSE: The purpose of this analysis was to determine the unique contribution of household income to the variance explained in psychological well-being (PWB) among a sample of colorectal cancer (CRC) survivors. METHODS: This study is a secondary analysis of data collected as part of the Health-Related Quality of Life in Long-Term Colorectal Cancer Survivors Study, which included CRC survivors with (cases) and without (controls) ostomies. The dataset included socio demographic, health status, and health-related quality of life (HRQOL) information. HRQOL was assessed with the modified City of Hope Quality of Life (mCOH-QOL)-Ostomy questionnaire and SF-36v2. To assess the relationship between income and PWB, a hierarchical linear regression model was constructed combining data from both cases and controls. RESULTS: After accounting for the proportion of variance in PWB explained by the other independent variables in the model, the additional variance explained by income was significant (R (2) increased from 0.228 to 0.250; P = 0.006). CONCLUSIONS: Although the study design does not allow causal inference, these results demonstrate a significant relationship between income and PWB in CRC survivors. The findings suggest that for non-randomized group comparisons of HRQOL, income should, at the very least, be included as a control variable in the analysis. PMID- 19132553 TI - Enhanced production of spinosad in Saccharopolyspora spinosa by genome shuffling. AB - Spinosad (spinosyns A and D) is a mixture of secondary metabolites produced by Saccharopolyspora spinosa. It is used in agriculture as a potent insect control agent with exceptional safety to non-target organisms. In this study, we applied genome shuffling of S. spinosa to achieve a rapid improvement of spinosad production. Ten strains with subtle improvements in spinosad production were obtained from the populations generated by the mutation with nitrosoguanidine and ultraviolet irradiation, and then they were subjected for recursive protoplast fusion. After four rounds of genome shuffling, a high yielding strain, designated as S. spinosa 4-7, was successfully isolated. Its production reached 547 mg/L, which was increased by 200.55% and 436.27% in comparison with that of the highest parent strain and the original strain, respectively. The subculture experiments indicated that the high producer of S. spinosa 4-7 was stable. Spinosad fermentation experiments by S. spinosa 4-7 were carried out in a 5-L fermentor, and its production of spinosad reached 428 mg/L after 168 h of fermentation. PMID- 19132554 TI - Substrate specificity and thermostability of the dehairing alkaline protease from Bacillus pumilus. AB - An alkaline protease (DHAP) from Bacillus pumilus has shown great potential in hide dehairing. To get better insights on its catalytic properties for application, the substrate specificity and thermostability were investigated using five natural proteins and nine synthetic peptides. The results showed that DHAP could hydrolyze five proteins tested here in different specificity. Collagen, a component of animal skin, was more resistant to hydrolysis than casein, fibrin, and gelatin. Among the synthetic peptides, the enzyme showed activity mainly with tetrapeptide substrates with the catalytic efficiency in order of Phe>Leu>Ala at P1 site, although k(m) value for AAVA-pN is much lower than that for AAPL-pN and AAPF-pN. With tripeptide substrates, smaller side-chain group (Gly) at P1 site was not hydrolyzed by DHAP. The enzyme showed good thermostability below 60 degrees C, and lost activity so quickly above 70 degrees C. The thermostability was largely dependent on metal ion, especially Ca(2+), although other ions, like Mg(2+), Mn(2+), and Co(2+), could sustain stability at certain extent within limited time. Cu(2+), Fe(2+), as well as Al(3+), did not support the enzyme to retain activity at 60 degrees C even in 5 min. In addition, the selected metal ions could coordinate calcium in improvement or destruction of thermostability for DHAP. PMID- 19132555 TI - Biodiesel production from various oils under supercritical fluid conditions by Candida antartica lipase B using a stepwise reaction method. AB - In this study, we evaluate the effects of various reaction factors, including pressure, temperature, agitation speed, enzyme concentration, and water content to increase biodiesel production. In addition, biodiesel was produced from various oils to establish the optimal enzymatic process of biodiesel production. Optimal conditions were determined to be as follows: pressure 130 bar, temperature 45 degrees C, agitation speed 200 rpm, enzyme concentration 20%, and water contents 10%. Among the various oils used for production, olive oil showed the highest yield (65.18%) upon transesterification. However, when biodiesel was produced using a batch system, biodiesel conversion yield was not increased over 65%; therefore, a stepwise reaction was conducted to increase biodiesel production. When a reaction medium with an initial concentration of methanol of 60 mmol was used and adjusted to maintain this concentration of methanol every 1.5 h during biodiesel production, the conversion yield of biodiesel was 98.92% at 6 h. Finally, reusability was evaluated using immobilized lipase to determine if this method was applicable for industrial biodiesel production. When biodiesel was produced repeatedly, the conversion rate was maintained at over 85% after eight reuses. PMID- 19132556 TI - Enzymatic saccharification and viscosity of sawdust slurries following ultrasonic particle size reduction. AB - Results in a previous study showed up to a 55% increase in saccharification rates when the initial particle size range decreased from 590 < x < 850 microm down to 33 < x < 75 microm. The smaller particle sizes also lowered the viscosity of the slurry 50-fold (for an equivalent initial solids concentration). In this study, ultrasonic irradiation was employed to further reduce the particle size of sawdust slurries below the ranges in the previous study in an attempt to further increase enzymatic saccharification rates and lower the slurry viscosity. Average particle sizes were reduced to less than 1 microm under the conditions tested. Surprisingly, the amount and rates of sugar released in this study with the approximately 1 microm particles is comparable (maximum glucose release of 30%) to, but no better than that seen for particle sizes in the range of 33 < or = x < or = 75 microm (maximum glucose release of 31%). Also surprisingly, the viscosity increased as the average particle sizes in the slurries decreased, which is opposite to the trend in the previous study. For example, there was an approximately threefold increase in the viscosity between unsonicated samples with a range of 10 < or = x < or = 75 microm and sonicated samples with a range of 0.05 < or = x < or = 12 microm. This is attributed to the variations in surface characteristics of the particles which were characterized here using X ray diffraction profiles and SEM pictures. PMID- 19132557 TI - Application of artificial neural network for yield prediction of lipase-catalyzed synthesis of dioctyl adipate. AB - In this study, an artificial neural network (ANN) trained by backpropagation algorithm, Levenberg-Marquadart, was applied to predict the yield of enzymatic synthesis of dioctyl adipate. Immobilized Candida antarctica lipase B was used as a biocatalyst for the reaction. Temperature, time, amount of enzyme, and substrate molar ratio were the four input variables. After evaluating various ANN configurations, the best network was composed of seven hidden nodes using a hyperbolic tangent sigmoid transfer function. The correlation coefficient (R2) and mean absolute error (MAE) values between the actual and predicted responses were determined as 0.9998 and 0.0966 for training set and 0.9241 and 1.9439 for validating dataset. A simulation test with a testing dataset showed that the MAE was low and R2 was close to 1. These results imply the good generalization of the developed model and its capability to predict the reaction yield. Comparison of the performance of radial basis network with the developed models showed that radial basis function was more accurate but its performance was poor when tested with unseen data. In further part of the study, the feedforward backpropagation model was used for prediction of the ester yield within the given range of the main parameters. PMID- 19132558 TI - Cerebrovascular complications of methamphetamine abuse. AB - OBJECTIVE: Methamphetamine is a stimulant widely abused in the United States. The objective of this study was to demonstrate an association of methamphetamine use and ischemic stroke, subarachnoid hemorrhage, and intracerebral hemorrhage and to further reveal the underlying vascular pathology using neuroimaging and pathology. METHODS: This was a retrospective study based on medical chart review of admissions to the neurovascular service of a tertiary care medical center from January 2003 to July 2007. Cases included patients who used methamphetamine as documented by history or urine toxicology screening. RESULTS: From 1,574 records, 30 cases were identified. The mean age of patients was 43 years and the discharge diagnoses included ischemic stroke, intracerebral hemorrhage, and subarachnoid hemorrhage. All subarachnoid hemorrhages were aneurysmal with the majority of the aneurysms located in the anterior circulation. The majority of strokes were located in the anterior circulation. In many cases, radiologic imaging confirmed arterial stenoses in the vascular distribution of the stroke. One patient who presented with ischemic stroke had severe atherosclerosis of bilateral common, internal, and external carotid arteries. On pathology, there was no evidence of inflammation or necrosis to suggest vasculitis as a possible etiology. CONCLUSIONS: Methamphetamine use is associated with ischemic stroke, intracerebral hemorrhage, and subarachnoid hemorrhage, especially among young patients. We showed no evidence that the ischemic stroke associated with methamphetamine use is due to an inflammatory etiology but may be due to a process of accelerated atherosclerosis. PMID- 19132560 TI - Teaching physical exam skills using web-based video. PMID- 19132561 TI - Patient interviewing behaviors. PMID- 19132562 TI - Clinical research in family medicine: published articles 2000 and 2005. PMID- 19132564 TI - Teaching learners about pediatric caries prevention. PMID- 19132559 TI - Recent advances in the treatment of systemic sclerosis. AB - Systemic sclerosis (SSc) is a chronic autoimmune disease with clinical manifestations resulting from immune activation, fibrosis development, and damage of small blood vessels. Our aim was to critically illustrate the available data on the new treatments proposed for SSc to provide a clinically oriented overview of the current evidence. PubMed was used for literature search using "scleroderma" and "therapy" to identify all articles published on indexed journals between 1972 and 2008. The search was limited to publications in English and produced a total of 3,441 references, which included 735 review articles. These citations were then screened for articles dealing with the most recent therapy options for SSc, and 214 articles were selected for evaluation and discussion. Methotrexate, cyclophosphamide, calcium channel blockers, angiotensin converting enzyme inhibitors, prostacyclin analogues, D-penicillamine, and extracorporeal photopheresis are the most widely studied treatments for SSc and were considered as practiced treatments. Other therapeutic approaches have been developed more recently and include endothelin receptor antagonists and phosphodiesterase-5 inhibitors for pulmonary arterial hypertension and peripheral vascular disease. High-dose immunosuppression and stem cell transplantation constitute a promising treatment and data from randomized controlled trials are awaited. Intravenous gamma globulins, mycophenolate mophetil, collagen tolerance induction, rituximab, fluoxetine, pirfenidone, relaxin, halofuginone, anti-TGF beta antibodies, and tyrosine kinase inhibitors awaits more solid data. The clinical management of patients with SSc remains a challenge and currently involves practiced and newly proposed therapeutic approaches. The disease pleiomorphism poses numerous difficulties to determine ideal outcomes to be used in clinical trials. PMID- 19132565 TI - Fallibility/Forgiveness. PMID- 19132567 TI - Zooming in to health ethics: an action to promote ethics. PMID- 19132568 TI - Student attitudes: potential barriers to implementing a community medicine field activity. AB - OBJECTIVE: Clerkship-year medical students may have complex attitudes regarding communities in which they train. Our objective was to assess medical students' attitudes following implementation of a community medicine experience in their clerkship year. METHODS: We analyzed transcripts of audiotaped medical student group discussions following a community field activity. A multidisciplinary team coded the transcripts using content analysis techniques to identify key features of student narratives regarding students' attitudes and attributions regarding their community medicine experiences. RESULTS: Students expressed several important perceptions regarding the community in which they conducted their field experience. These included strong insider/outsider themes, resentment that historical legacy shape the doctor-patient relationship, and concerns over personal safety and educational relevance of the activity. Some students noted improvement in understanding of patients within their communities. CONCLUSIONS: Negative attitudes and attributions expressed by students can act as obstacles in the development of community medicine initiatives, hinder professional development if unaddressed, and have the potential to contribute to health disparities. Further work on how to incorporate community medicine training is needed. PMID- 19132569 TI - First-year medical students can demonstrate EHR-specific communication skills: a control-group study. AB - BACKGROUND AND OBJECTIVES: Graduating medical students will universally use electronic health records (EHRs), yet a June 2007 literature search revealed no descriptions of EHR-specific communication skills curricula in US medical schools. We designed and tested methods to teach first-year medical students to optimally integrate EHRs into physician-patient communication in ambulatory encounters. METHODS: We randomly assigned 17 volunteer students to control (n=8) and intervention (n=9) groups. Both groups learned the mechanics of documenting patient histories using the EHR. Additionally, we taught the intervention group EHR-specific communications skills using guided discovery, brief didactics, and practice role plays. We compared both groups' general and EHR-specific communications skills using a standardized patient (SP) case. RESULTS: Students receiving EHR communication skills training performed significantly better than controls in six of 10 EHR communication skills. In 10 of 11 general communication skills, there were no significant differences between groups. CONCLUSIONS: First year medical students can demonstrate EHR communication skills early in their medical training. However, in our setting, students did not spontaneously demonstrate EHR skills without instruction, and such skills did not correlate with general communication skills. PMID- 19132570 TI - The central city site: an urban underserved family medicine training track. AB - BACKGROUND AND OBJECTIVES: We describe the development of an urban track in family medicine residency designed to recruit a high percentage of minority students and promote their future practice in urban, underserved areas of Milwaukee. We report here on the residents and their first practice location and compared this information to what occurred in our original "main" residency program. METHODS: Information about the program's development was obtained through testimonials from faculty and residency graduates and review of the original accreditation application to the Residency Review Committee. Information about the residents and their practice locations was obtained from the National Resident Matching Program and graduate placement data. RESULTS: The goal of training more minority doctors in Milwaukee was met, with eight of 16 (50%) residents at our urban-track site from minority groups. This compared to only 12% at our main program. Thirty-eight percent of graduates stayed to practice in an underserved area, compared to only 21% in our main program. CONCLUSIONS: Development of an urban track for our family medicine residency increased the number of minority physicians trained and the number of physicians practicing in underserved areas after graduation. PMID- 19132571 TI - A comparison of sodium phosphosoda purgative to polyethylene glycol bowel preparations prior to colonoscopy. AB - BACKGROUND AND OBJECTIVES: Successful colonoscopy is contingent upon adequate bowel preparation, which is often achieved using either an oral sodium phosphate preparation or a polyethylene glycol-based preparation. Comparison of the relative performance of these two classes of agents has been assessed only in the context of clinical trials (and with mixed findings). However, efficacy measured in clinical trials often is not reflective of effectiveness in clinical practice. We undertook this analysis to determine the relative clinical effectiveness of oral sodium phosphosoda versus polyethylene glycol in clinical practice. METHODS: Subjects (n=343) were selected from among patients receiving outpatient colonoscopy at our institution between January 2004 and February 2006. Demographic, biochemical, and comorbid disease data were abstracted from the electronic medical record. Colonoscopy preparation, indication, and preparation quality were abstracted from colonoscopy reports. RESULTS: As compared to subjects receiving polyethylene glycol, those receiving oral sodium phosphosoda had an adjusted odds ratio (OR) (95% confidence intervals [CIs]) for adequate/good/excellent bowel preparation quality of 2.23 (1.18--4.22) and an adjusted OR (95% CIs) for good/excellent bowel preparation of 2.24 (1.26--3.97). There was no interaction on the basis of colonoscopy indication. CONCLUSIONS: Oral sodium phosphate-based purgatives were associated with significantly better bowel preparation quality among outpatients at our center. PMID- 19132572 TI - Decreased continuity in a residency clinic: a consequence of open access scheduling. AB - BACKGROUND AND OBJECTIVES: Open access scheduling decreases waiting time to see physicians by using same-day appointment scheduling. In primary care residency training, continuity of care may be difficult to preserve with this method of scheduling because requirements for rotations often results in residents being unavailable in their primary clinic practice. Our objective was to examine continuity of care in a family medicine residency clinic during a 1-year period prior to implementation of open-access scheduling and during a 1-year period after open access scheduling started. METHODS: Two indices to measure continuity were used: the Usual Provider Continuity Index (UPC) and the Modified Modified Continuity Index (MMCI). The Mann-Whitney test was used to determine differences in the UPC and MMCI between groups. RESULTS: The mean UPC and MMCI scores decreased with open access scheduling. Mean UPC was 0.59 with traditional scheduling versus 0.55 with open access scheduling. Mean MMCI was 0.51 for traditional scheduling and 0.44 with open access. CONCLUSIONS: Continuity of care decreased in our clinic after implementation of open access scheduling. Our results have implications for all primary care residency training programs since one of the hallmarks of primary care is maintaining continuity in the physician patient relationship. PMID- 19132573 TI - The effect of a carve-out advanced access scheduling system on no-show rates. AB - BACKGROUND AND OBJECTIVES: The relationship between advanced access scheduling and no-show rates in academic settings is unclear. The purpose of this analysis is to assess the effect of moving to a carve-out model of an advanced access scheduling system on no-show rates in an academic practice. METHODS: A multivariable logistic regression with repeated measures analysis of patient visits was used to assess the relationship between the time to third-next available appointment and no-show rates. RESULTS: The time to third-next available appointment was not a significant factor in appointment-keeping behavior. Rather, provider-patient continuity, wait time for an appointment, the number of previous appointments, and type of provider were strong predictors of a patient no-show. CONCLUSIONS: Implementation of the concepts of advanced access scheduling does not directly result in a reduction in no-show rates; rather, provider-patient continuity, wait time to an appointment, and other factors may be a more important measure. PMID- 19132575 TI - International Family Medicine Education. PMID- 19132574 TI - Advanced access, open access, and continuity of care: should we enforce continuity? PMID- 19132578 TI - Evidence for age-related changes to temporal attention and memory from the choice time production task. AB - The effect of aging on interval timing was examined using a choice time production task, which required participants to choose a key response based on the location of the stimulus, but to delay responding until after a learned time interval. Experiment 1 varied attentional demands of the response choice portion of the task by varying difficulty of stimulus-response mapping. Choice difficulty affected temporal accuracy equally in both age groups, but older participants' response latencies were more variable under more difficult response choice conditions. Experiment 2 tested the contribution of long-term memory to differences in choice time production between age groups over 3 days of testing. Direction of errors in time production between the two age groups diverged over the 3 sessions, but variability did not differ. Results from each experiment separately show age-related changes to attention and memory in temporal processing using different measures and manipulations in the same task. PMID- 19132579 TI - Preliminary study on intrasplenic implantation of artificial cell bioencapsulated stem cells to increase the survival of 90% hepatectomized rats. AB - We implanted artificial cell bioencapsulated bone marrow mesenchymal stem cells into the spleens of 90% hepatectomized (PH) rats. The resulting 14 days survival rate was 91%. This is compared to a survival rate of 21% in 90% hepatectomized rats and 25% for those receiving free MSCs transplanted the same way. Unlike free MSCs, the bioencapsulated MSCs are retained in the spleens and their hepatotrophic factors can continue to drain directly into the liver without dilution resulting in improved hepatic regeneration. In addition, with time the transdifferentiation of MSCs into hepatocyte-like cells in the spleen renders the spleen as a ectopic liver support. PMID- 19132580 TI - Higher order factor structure of the WISC-IV in a clinical neuropsychological sample. AB - A confirmatory factor analysis was conducted examining the higher order factor structure of the WISC-IV scores for 344 children who participated in neuropsychological evaluations at a large children's hospital. The WISC-IV factor structure mirrored that of the standardization sample. The second order general intelligence factor (g) accounted for the largest proportion of variance in the first-order latent factors and in the individual subtests, especially for the working memory index. The first-order processing speed factor exhibited the most unique variance beyond the influence of g. The results suggest that clinicians should not ignore the contribution of g when interpreting the first-order factors. PMID- 19132581 TI - Challenges in researching life with HIV/AIDS: an intersectional analysis of black African migrants in London. AB - Most social science research on HIV has focused on prevention. The arrival of new therapies generated more studies on life with HIV. However most have been carried out in developed world contexts. Much less is known about the vast majority of those living with HIV and dying from AIDS. If this gap is to be filled, more qualitative research will be needed on affected individuals in the developing world and also among migrants who have left developing countries to live in the diaspora. It will also be essential to explore the lives of individuals from the same communities who may experience HIV in very different ways as a result of their gender and/or sexuality. This paper presents findings from three studies of Black African migrants living with HIV in London. It uses an intersectional approach to examine the similarities and the differences between the experiences of heterosexual women, heterosexual men and gay and/or bisexual men. The article highlights the importance of research of this kind both for providing the evidence base for context-specific policy development and also for making better conceptual and theoretical sense of the impact of HIV on individuals and their lives. PMID- 19132582 TI - South African teachers' reflections on the impact of culture on their teaching of sexuality and HIV/AIDS. AB - This paper aims to explore South African Life Orientation teachers' perception and practice of teaching HIV/AIDS and sexuality in a cultural perspective. We aim to investigate how teachers respond to perceived cultural differences between the local community and the content of their teaching. Data were collected through interviews with teachers who taught students in grades 8 or 9 in public high schools. The teachers expressed differing viewpoints regarding the rationale for teaching about HIV/AIDS and sexuality. Many teachers saw teaching these topics as a response to declining moral standards, while others suggested that they were teaching issues that parents failed to address. The teachers were more concerned about young people's sexual behaviour than about preventing HIV/AIDS. They perceived that cultural contradictions between what was taught and local cultural values were an issue to which they needed to respond, although they differed in terms of how to respond. Some took an adaptive approach to try to avoid conflicts, while others claimed the moral neutrality of their teaching. Teaching about sexuality was perceived to be challenging in terms of language and communication norms. Teaching about HIV/AIDS was perceived as challenging because teachers often needed to convince students about the reality of AIDS. PMID- 19132583 TI - Degradation of RDX using granular iron and nickel-plated granular iron. AB - Using granular iron (Fe) and nickel-plated iron (Ni/Fe), this paper examines the effectiveness of these two types of reactive materials for the treatment of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), a common groundwater and soil contaminant at military facilities. RDX degraded very rapidly in the presence of both Fe and Ni/Fe in column and batch experiments. Enhancement by Ni/Fe did not prove to be effective as the half-lives of RDX ranged from 3 to 24 seconds and 3 to 11 seconds in the Fe and Ni/Fe columns, respectively. Reaction vessel experiments and estimation of the mass transfer coefficient in the column indicated that reaction kinetics was mass transfer limited. Detailed analyses of reaction intermediates and products suggest that RDX degradation proceeds through direct electron transfer processes and following to the same pathways in the presence of Fe and Ni/Fe. The formation of carbon-containing products, including formaldehyde (up to 60%), CO2 (up to 45%) and formic acid (1%) and the nitrogen containing products of ammonium (up to 48%) and N2O (up to 13%), provides convincing evidence that RDX was completely decomposed to non-toxic end products. CO2, previously reported to form only in biological or Fe-microbial combined systems, was detected as one of the main C-bearing end product. Therefore, this study shows that Fe is an effective material for remediating groundwater and industrial effluents containing RDX; and the use of additional enhancement, either biological or with Ni catalyst, does not provide additional advantages. PMID- 19132584 TI - Effect of ozone and granular activated coal (GAC) on the bioactivity of drinking water. AB - In this research, the appearance of easily biodegradable organic material in ozonation and granular activated coal (GAC) filtration was studied. The amount of bioactivity was measured by conventional AOC analyses used in two different modes and also using quite a new growth potential (GP) method. GAC filtration without ozone doubled the amount of AOC of the chemically treated surface water, whereas by ozonation with GAC filtration it was possible to halve the amount of the AOC. The measurement of GP was noticeably simpler than measuring AOC, but for wider use more parallel studies are needed for the comparability of the results of the analysis. PMID- 19132585 TI - Preliminary water quality assessment of Spunky Bottoms restored wetland. AB - The approximately 1200-acre "Spunky Bottoms" wetland in Southern Illinois has been undergoing restoration to conditions prior to levying of the Illinois River and draining of adjacent floodplain for intensive agriculture (circa 1900). As part of a long-term water quality impact assessment of this restoration project, baseline water quality monitoring was conducted soon after restoration began. During this baseline/preliminary assessment, water samples were taken every 2-4 weeks from 10 sampling wells and seven surface water sites throughout the wetlands area for a period of 18 months. Measured parameters include nutrients (nitrate (NO3-) and phosphate (PO4(3-)), cations and anions (SO4(2-), Cl-, Na+, K+, Mg2+, Ca2+) commonly found in surface and well water, trace metals (Al, Cd, Cu, Fe, Mn, Ni, Pb, Se, Zn), total dissolved solids (TDS), pH, and trace organics (triazine herbicides and their metabolites). In general, highest concentrations of ions were found in the southwest and northeast perimeter of the wetland area for both surface and ground water samples. Primarily low concentrations of heavy metals and organic compounds were found throughout the wetland sampling area. Distribution of NO3--N suggests that this restored wetland, even at its infant age, may still contribute to biogeochemical (particularly N) element cycling. Continued monitoring and further research is necessary to determine long-term specific contribution of restored wetland to biogeochemical cycles. PMID- 19132586 TI - Cadmium concentration in sea bottom sediment and its potential risk in the upper Gulf of Thailand. AB - Untreated or partially treated waste water discharge from industrial and domestic sources entering the Upper Gulf of Thailand have been reported to increase cadmium concentration in bottom sediment. This study was directed at providing a better understanding of cadmium transformation in the sediment from the area. Sediment samples collected from Chao Phraya River mouth (CPY), Bang Pakong River mouth (BPK) and Klong Dan estuary (KD) located in the Upper Gulf of Thailand were analyzed for cadmium concentration in various sediment particle size fractions. Using laboratory microcosms, cadmium release from sediment as affected by salinity and sediment redox condition was studied. A higher concentration of cadmium (0.2-0.6 microg/g dry weight) was measured in finer sediment particle size fractions (<0.075 mm) as compared to courser fractions at all sampling stations. Cadmium release from the sediment to water was influenced by both salinity and redox condition. Sediment was spiked with 10 ppm cadmium which is the cadmium level in sediment adopted by the Australia and New Zealand sediment quality guidelines which can cause adverse environmental impacts. Experiments conducted showed soluble cadmium concentration at sediment oxidation reduction conditions representative of bottom sediment were at levels that can adversely impact aquatic organisms, according to the PCADMIUM water quality guideline. In Thailand, there is no sediment quality guideline. Based on these experiments, 10 ppm of cadmium in sediment was recommended as a regulatory guidelines for allowable levels of cadmium in sediment in the study area. PMID- 19132587 TI - Evaluation of fecal contamination indicators (fecal coliforms, somatic phages, and helminth eggs) in ryegrass sward farming. AB - The effect of soil supplementation with biosolids at various ratios on fecal origin microorganism activity was evaluated in a ryegrass sward farm. Fifteen plots with 3 different soil and biosolid mixture ratios were assessed. Soil and grass were sampled over a period of 4 months (days 0, 30, 45, 60, 75, and 120) for soil and on days 75 and 120 for grass, corresponding to first and second grass harvest periods. We analyzed fecal coliforms, somatic phages, helminth eggs, and environmental factors, such as rainfall, temperature, and moisture. The fecal coliforms decreased by 2 logarithmic units (LU) in all soils containing biosolids and by 1 LU in the soil alone and in biosolid control plots alone. The concentration of somatic phages decreased to 2 to 3 LU in the soil containing biosolids and to 1 to 2 LU in the control plots. In contrast, however, there was a noticeable increase in helminth eggs on days 75 ad 120, but not in the soil control alone. Maximum concentrations (10(2) CFU/g TS; colony forming units per gram total solids) of fecal coliforms were found on the grass and in other samples, but the concentrations of phages and helminth eggs were below detection limits. Environmental factors did not significantly influence the results, and grass production increased from 35 to 50 Ton/Ha (tons per hectare) with biosolid supplementation, as compared with controls (14 Ton/Ha). PMID- 19132588 TI - Removal of ethylene and bioaerosol by chlorine dioxide using a chemical scrubbing system in a fruit and vegetable storage facility. AB - Ethylene (C2H4) and bioaerosol are commonly present in the inside atmosphere of postharvest fruit and vegetable storage facilities, which may affect the aging of postharvest fruit and human health. We have assessed the feasibility of chlorine dioxide (ClO2) as the scrubbing solution in a chemical scrubbing tower for simultaneously removing C2H4 and bioaerosol emissions from a gas stream. Parameters such as the ClO2concentration, contact time, and liquid-to-gas (L/G) ratio were examined with the aim of determining the optimal operating conditions. Using the system reported here, the optimal C2H4 removal efficiency was 99.5% when 500 ppm ClO2 was used at a reaction time of 30-60 s under a continuous non recycle ClO2 flow mode. In terms of C2H4 removal, a greater L/G resulted in a higher C2H4 removal efficiency up to the optimal ratio of 12.5. In terms of the simultaneous removal of C2H4 and bioaerosol, the removal efficiency of C2H4 was 99.2% and those for the bioaersols of Escherichia coli and Staphylococcus aureus were 99.92 and 99.10%, respectively, under a continuous non-recycle flow mode. Our results also indicate that oxidation reduction potential (ORP) can be a valuable indicator for the timing of the replacement of the scrubbing solution in the system under a continuous recycle flow mode. Additional confirmation of the feasibility of the ORP as an indicator of C2H4 and bioaerosol removal in situ was obtained in a 3-month test of our system in continuous recycle flow mode with the periodical replacement of scrubbing solution, ClO2. The removal efficiencies for C2H4, bacterial and fungus aerosol, and total hydrocarbon compounds (THC) were 83.4, 96.8, 96.1, and 76.5%, respectively. Our results prove that ClO2 is an excellent scrubbing solution in the chemical scrubbing tower for the removal of C2H4 emissions and bioaerosol. We demonstrate, for the first time, the feasibility of this system in a fruit and vegetable storage facility. PMID- 19132589 TI - Behaviour of fixed-bed column for the adsorption of malachite green on surfactant modified alumina. AB - Prepared surfactant-modified alumina (SMA) was used to remove malachite green (MG) from aqueous media. At a dose of 10 g/L, SMA removed approximately 99% MG (initial concentration 100 mg/L). The adsorption capacity (Qmax) of SMA was 185 mg/g as calculated from Langmuir isotherm. In a fixed-bed column study, using the MG-spiked distilled water, the column design parameters were evaluated by Logit model at a bed depth of 10 cm. The adsorption rate constant (K) and adsorption capacity (No) was obtained as 0.002636 L/(mg h) and 76283.16 mg/L for the minimum bed depth 3.33 cm in the 1st cycle. Acetone was used for desorption of MG from SMA. In batch regeneration study, regenerated SMA could remove only approximately 80% of MG under the same experimental conditions. In column regeneration study, the efficiency of the regenerated bed decreased and the values obtained as, K=0.007931 L/mg h and No=12341.08 mg/L for the minimum bed depth of 6.83 cm. Column study was conducted with the real MG bearing wastewater (MG concentration was 396.54 mg/L) under the same experimental condition. The value of adsorption rate constant (K) and adsorption capacity (No) was obtained as 0.0008786 L/(mg h) and 197939.02 mg/L, respectively for the minimum bed depth 5.92 cm. PMID- 19132590 TI - Citotoxicity status of electroplating wastewater prior/after neutralization/purification with alkaline solid residue of electric arc furnace dust. AB - Toxicological safety of new procedure for the neutralisation/purification of wastewater originated from zinc plating facility was investigated. Wastewater was treated with alkaline solid residue-by-product of zinc recovery from electric arc furnace dust. For determination of cytotoxic potential of untreated and purified wastewater MTT test on HEp2 (human laryngeal carcinoma) and HeLa (human cervical carcinoma) cells lines and alkaline comet assay on human leukocytes were used. Then 100% of the sample as well as different dilutions were tested. Compared to negative control 100, 75 and 50% of the sample of untreated wastewater significantly decreased survival of both HEp2 and HeLa cell lines. In the presence of undiluted sample survival percentage of HeLa and HEp2 cells were only 2.3 and 0.3% respectively. Only undiluted purified wastewater showed slight but insignificant decrease of the survival of both cell lines. Even 0.5% of the sample of original electroplating wastewater exhibited significantly higher value of all comet assay parameters compared to negative control. There was no significant difference between negative control and purified wastewater for any of comet assay parameters. Significantly lower level of primary DNA damage recorded after treatment with purified water, even comparable with negative control, confirmed effectiveness of the purification process. PMID- 19132591 TI - In vitro assessment of genotoxic effects of electric arc furnace dust on human lymphocytes using the alkaline comet assay. AB - In vitro genotoxic effects of leachates of electric arc furnace dust (EAFD) on human peripheral lymphocytes, assessed prior and following the treatment with a strong alkaline solution were investigated using the alkaline comet assay. Prior and following the treatment, lymphocytes were incubated with leachate of EAFD for 6 and 24 hours at 37 degrees C. Negative controls were also included. Mean values of the tail lengths established in the samples treated with the leachate stemming from the original dust for 6 and 24 hours, were 15.70 microm and 16.78 microm, respectively, as compared to 12.33 microm found in the control sample. Slight, but significant increase in the tail length was also found with the dust treated with a strong alkaline solution (13.37 microm and 13.60 microm). In case of high heavy metal concentrations (the extract of the original furnace dust), the incubation period was revealed to be of significance as well. The obtained results lead to the conclusion that alkaline comet assay could be used as a rapid, sensitive and low-cost tool when assessing genotoxicity of various waste materials, such as leachates of the electric arc furnace dust. PMID- 19132592 TI - A molecular fragments variable connectivity index for studying the toxicity (Vibrio fischeri pT50) of substituted-benzenes. AB - Molecular fragments variable connectivity index (mfVCI) has been proposed as a molecular descriptor, which has been applied to the study of toxicity (Vibrio fischeri pT50) of 138 substituted-benzenes. Molecular fragments are defined as the atoms or functional groups having different characteristics due to different chemical bonding, which are considered as the molecular structure unit for the major influence of toxicity. The molecular fragment is regarded as the vertex of the topological diagram and endue with variable wi as substitute for the topological matrix diagonal. The QSAR model obtained by molecular fragments variable connectivity index shows good descriptive (R2=0.88) and predictive (Q2EXT=0.72) performance. The optimal weights of the molecular fragments have the ability to interpret the mechanism from the compounds' insight structure. PMID- 19132593 TI - Modeling of catalytic ozonation process in a three-phase reactor. AB - In this research, the main objective was to determine the flow characteristics of a three-phase reactor in order to use this knowledge in the modeling of catalytic ozonation of aqueous dye solutions. Therefore, the stimulus-response method was used in the tracer experiments; thus, the degree of liquid mixing in the reactor was estimated by means of residence time distribution, Peclet number and axial dispersion coefficient in the presence and the absence of the catalyst. Experimental data were obtained by performing the catalytic ozonation of aqueous Acid Red-151(AR-151) and Remazol Brilliant Blue-R (RBBR) dye solutions, in the presence of perfluorinated-octyl-alumina (PFOA) catalyst particles at different operating conditions. The chemical oxygen demand (COD), the dye and ozone concentrations in the liquid phase were measured at the steady state along the height of the column reactor and at the exit. According to the results, it was observed that the gas-liquid reactor without the catalyst particles showed a hydrodynamic behavior equivalent to two or three completely stirred tank reactors (CSTRs) in-series for the conventional ozonation process. The presence of catalyst particles caused the flow behavior of the three phase reactor to approach to one CSTR or two CSTRs in-series depending on the gas and liquid flow rates so that the modeling of the catalytic ozonation process was done satisfactorily on that basis. The modeling results showed satisfactory agreement with the experimental ones in the prediction of outlet dye and dissolved ozone concentrations from the reactor, especially at relatively high gas velocities (QG=150 and 200 L h(-1)) for AR-151, where the dissolved ozone concentration was not limited. However, the discrepancy was about 15% between the theory and experiment at the lower gas flow rates due to the limited ozone concentrations with respect to the dye concentrations at the high inlet dye concentration of AR 151 (CD,i=100 mg L(-1)). For RBBR, the predicted and experimental values were very close to each other since ozone limitation or dye abundance was not realized in these experiments. PMID- 19132594 TI - Treatability of atrazine in a simulated DEPHANOX process. AB - In this study a simulated DEPHANOX process was used, including anaerobic/anoxic and oxic phases. An upflow anaerobic sludge blanket reactor (UASB), an anoxic sludge blanket reactor (UANSB) and a completely stirred tank reactor (CSTR) were used, sequentially. The atrazine, chemical oxygen demand (COD) removal efficiencies, methane and nitrogen (N2) gas productions and volatile fatty acid (VFA) concentrations were monitored to assess the stability and the performance of anaerobic/anoxic and oxic reactors. The produced intermetabolites were Diaminochloroatrazine(DACT), Desethylatrazine(DEA), Deisopropylatrazine(DIA) urea, ammonia, aromatic amines, Cl(-1) and NO3-N. For maximum atrazine and COD removal efficiencies (86% and 82%, respectively) the optimum atrazine concentrations were between 30-80 mg L(-1). The methane gas percentage varied between 40 and 68% while no N2 production was observed in the anaerobic UASB reactor; 6-10 mg L(-1) of urea, 4-21 mg L(-1) of ammonia, 8-10 mg L(-1) of aromatic amine and 4-6 mg L of Cl(-1) were detected during anaerobic atrazine degradation. 25-45% N2 gas production was observed in the anoxic reactor while the methane gas production was 1-5%. In the aerobic phase COD and atrazine were removed with removal efficiencies of 98% and 99% for initial atrazine concentrations of 0.3 mg L(-1) and 2.5 mg L(-1), respectively. PMID- 19132596 TI - The irre cell recognition module (IRM) proteins. AB - One of the most challenging problems in developmental neurosciences is to understand the establishment and maintenance of specific membrane contacts between axonal, dendritic, and glial processes in the neuropils, which eventually secure neuronal connectivity. However, underlying cell recognition events are pivotal in other tissues as well. This brief review focuses on the pleiotropic functions of a small, evolutionarily conserved group of proteins of the immunoglobulin superfamily involved in cell recognition. In Drosophila, this protein family comprises Irregular chiasm C/Roughest (IrreC/Rst), Kin of irre (Kirre), and their interacting protein partners, Sticks and stones (SNS) and Hibris (Hbs). For simplicity, we propose to name this ensemble of proteins the irre cell recognition module (IRM) after the first identified member of this family. Here, we summarize evidence that the IRM proteins function together in various cellular interactions, including myoblast fusion, cell sorting, axonal pathfinding, and target recognition in the optic neuropils of Drosophila. Understanding IRM protein function will help to unravel the epigenetic rules by which the intricate neurite networks in sensory neuropils are formed. PMID- 19132597 TI - Working memory in bees: also in flies? AB - Decision making requires reference to both actual and remote information in the context of the requirements of the animal. Here, I explore a cognitive approach to decision making in honeybees and ask the question whether flies may share the faculties observed in bees. Examples are drawn primarily from natural behavior in bees, particularly from navigation and communication. I conclude that studies in Drosophila learning and memory in the tradition of Martin Heisenberg may gain from cognitive concepts, meaning that the "internal doing" of the brain should be included in the search for the neural basis of decision making. PMID- 19132598 TI - The Wuerzburg hybridoma library against Drosophila brain. AB - This review describes the present state of a project to identify and characterize novel nervous system proteins by using monoclonal antibodies (mAbs) against the Drosophila brain. Some 1,000 hybridoma clones were generated by injection of homogenized Drosophila brains or heads into mice and fusion of their spleen cells with myeloma cells. Testing the mAbs secreted by these clones identified a library of about 200 mAbs, which selectively stain specific structures of the Drosophila brain. Using the approach "from antibody to gene", several genes coding for novel proteins of the presynaptic terminal were cloned and characterized. These include the "cysteine string protein" gene (Csp, mAb ab49), the "synapse-associated protein of 47 kDa" gene (Sap47, mAbs nc46 and nb200), and the "Bruchpilot" gene (brp, mAb nc82). By a "candidate" approach, mAb nb33 was shown to recognize the pigment dispersing factor precursor protein. mAbs 3C11 and pok13 were raised against bacterially expressed Drosophila synapsin and calbindin 32, respectively, after the corresponding cDNAs had been isolated from an expression library by using antisera against mammalian proteins. Recently, it was shown that mAb aa2 binds the Drosophila homolog of "epidermal growth factor receptor pathway substrate clone 15" (Eps15). Identification of the targets of mAbs na21, ab52, and nb181 is presently attempted. Here, we review the available information on the function of these proteins and present staining patterns in the Drosophila brain for classes of mAbs that either bind differentially in the eye, in neuropil, in the cell-body layer, or in small subsets of neurons. The prospects of identifying the corresponding antigens by various approaches, including protein purification and mass spectrometry, are discussed. PMID- 19132599 TI - Contact. PMID- 19132600 TI - From form to function: the ways to know a neuron. AB - The shape of a neuron, its morphological signature, dictates the neuron's function by establishing its synaptic partnerships. Here, we review various anatomical methods used to reveal neuron shape and the contributions these have made to our current understanding of neural function in the Drosophila brain, especially the optic lobe. These methods, including Golgi impregnation, genetic reporters, and electron microscopy (EM), necessarily incorporate biases of various sorts that are easy to overlook, but that filter the morphological signatures we see. Nonetheless, the application of these methods to the optic lobe has led to reassuringly congruent findings on the number and shapes of neurons and their connection patterns, indicating that morphological classes are actually genetic classes. Genetic methods using, especially, GAL4 drivers and associated reporters have largely superceded classical Golgi methods for cellular analyses and, moreover, allow the manipulation of neuronal activity, thus enabling us to establish a bridge between morphological studies and functional ones. While serial-EM reconstruction remains the only reliable, albeit labor intensive, method to determine actual synaptic connections, genetic approaches in combination with EM or high-resolution light microscopic techniques are promising methods for the rapid determination of synaptic circuit function. PMID- 19132602 TI - Relations among religiousness, spirituality, and sexual practices. AB - Although religiousness is commonly associated with limited sexual activity, little is known about spirituality's unique effect. Study aims involved measuring spirituality's unique affect on young adults' sexual practices (frequency of sex, number of sexual partners, and condom use) and determining whether spirituality adds significant increment over well-established predictors. Three hundred fifty three (61% female) heterosexual young adults aged 17 to 29 completed this cross sectional, self-report questionnaire study. Hierarchical regression analyses demonstrated spirituality is positively associated with participants' number of sexual partners and frequency of sex without a condom. Spirituality contributed to the prediction of participants' number of partners and condom use above and beyond the variance accounted for by religiousness, alcohol use, and impulsivity. A moderating effect for gender was found. Spirituality appears to have a unique and strong association with the sexual practices of young adults, particularly women, and should be assessed in future studies of young adults' sexual practices. PMID- 19132603 TI - Attitudes toward stereotypical versus counterstereotypical gay men and lesbians. AB - This study examined differences in the way heterosexual men and women feel about gay men and lesbians who either confirm or disconfirm prevailing stereotypes. Fifty-three heterosexual college men and women read descriptions about 2 fictitious homosexual students at their university, both of whom were either male or female. One of the students was portrayed as being more stereotypically masculine, whereas the other was portrayed as more stereotypically feminine. Participants rated the targets on a variety of dimensions, including liking. Male participants liked the masculine gay man more than the feminine gay man, whereas female participants showed similar degrees of liking for both men. Both male and female participants reported greater liking for the feminine lesbian than the masculine lesbian. Implications for understanding attitudes toward stereotypical versus counterstereotypical gay men and lesbians are discussed. PMID- 19132601 TI - Swiss cheese et allii, some of the first neurodegenerative mutants isolated in Drosophila. AB - Drosophila has only recently become a model organism to study progressive neurodegeneration, mainly using transgenic flies expressing human disease genes. However, classical forward genetics isolating and characterizing fly mutants that show characteristic features of progressive neurodegeneration can also provide a useful tool to get insights into the mechanisms of neurodegeneration. Interestingly, the first such mutants have been already isolated in the 1970s, and this review focuses on the description of four such mutants originally isolated by Martin Heisenberg. PMID- 19132604 TI - Does post-event cognitive load undermine thought suppression and increase intrusive memories after exposure to an analogue stressor? AB - Ironic process theory has been used in part to explain the phenomenon of intrusive memories in various disorders including post-traumatic stress disorder. How thought suppression interacts with other cognitive processes believed to be instrumental in the development of traumatic intrusive memory was tested. In an analogue design 120 participants were randomised to five conditions, four of which also required participants to attempt to suppress intrusive memories after viewing a film of traumatic content. Participants in three conditions were also required to perform concurrent tasks that acted as a cognitive load during suppression. Intrusive memories were recorded during the experimental phase and at 1-week follow-up. Contrary to predictions, post-film processing did not undermine suppression success. There was some suggestion that post-film processing resulted in those participants experiencing intrusions of shorter duration than the no-suppression control group in two 5-minute intrusion monitoring intervals at the initial and follow-up phase of the experiment, but this was not reflected in a 1-week diary measure of intrusions. All experimental groups performed in a similar fashion in terms of memory testing of the film's content. The findings are discussed in the context of ironic process theory and cognitive models of post-traumatic stress. PMID- 19132605 TI - The use of autobiographical knowledge in age estimation. AB - Event dates are not directly associated with memories, so the processes by which we maintain a sense of time and sequence in our autobiographical memories is of considerable interest. The present study examined participants' reported age estimation strategies for childhood memories retrieved using a Galton-Crovitz cueing technique. The results indicate that all three categories of autobiographical knowledge in Conway and Pleydell-Pearce's (2000) self-memory system model-lifetime periods, general events, and event-specific details-support temporal inferences. However, participants most frequently used lifetime period knowledge to provide an initial age range, and event-specific knowledge was used to confirm or narrow the range of their estimated age. PMID- 19132607 TI - Internationalism and global norms for neuroethics. PMID- 19132608 TI - Responsibility and speculation: on possible applications of pediatric fMRI. PMID- 19132610 TI - BIID-Aqua Fortis for scientific explanations of psychic phenomena? PMID- 19132609 TI - Ethical challenges and interpretive difficulties with non-clinical applications of pediatric FMRI. AB - In this article, we critically examine some of the ethical challenges and interpretive difficulties with possible future non-clinical applications of pediatric fMRI with a particular focus on applications in the classroom and the courtroom - two domains in which children come directly in contact with the state. We begin with a general overview of anticipated clinical and non-clinical applications of pediatric fMRI. This is followed by a detailed analysis of a range of ethical challenges and interpretive difficulties that trouble the use of fMRI and are likely to be especially acute with non-clinical uses of the technology. We conclude that knowledge of these challenges and difficulties should influence policy decisions regarding the non-clinical uses of fMRI. Our aim is to encourage the development of future policies prescribing the responsible use of this neuroimaging technology as it develops both within and outside the clinical setting. PMID- 19132611 TI - The fear of new technology: a naturally occurring phenomenon. PMID- 19132612 TI - What we should really worry about in pediatric functional magnetic resonance imaging (FMRI). PMID- 19132613 TI - Considerations for pediatric neuroimaging at the translational coalface. PMID- 19132614 TI - Ethical and regulatory issues in pediatric research supporting the non-clinical application of fMR imaging. PMID- 19132615 TI - Enthusiasm for functional magnetic resonance imaging (FMRI) often overlooks its dependence on task selection and performance. PMID- 19132616 TI - Children in non-clinical functional magnetic resonance imaging (FMRI) studies give the scan experience a "thumbs up". PMID- 19132617 TI - A neuroimaging perspective on the use of functional magnetic resonance imaging (FMRI) in educational and legal systems. PMID- 19132618 TI - Functional magnetic resonance imaging (FMRI) in the classroom. PMID- 19132619 TI - A holistic approach to the use of pediatric FMRI. PMID- 19132620 TI - Considering the long term in the short term use of FMRI in the classroom. PMID- 19132621 TI - Body integrity identity disorder (BIID)--is the amputation of healthy limbs ethically justified? AB - The term body integrity identity disorder (BIID) describes the extremely rare phenomenon of persons who desire the amputation of one or more healthy limbs or who desire a paralysis. Some of these persons mutilate themselves; others ask surgeons for an amputation or for the transection of their spinal cord. Psychologists and physicians explain this phenomenon in quite different ways; but a successful psychotherapeutic or pharmaceutical therapy is not known. Lobbies of persons suffering from BIID explain the desire for amputation in analogy to the desire of transsexuals for surgical sex reassignment. Medical ethicists discuss the controversy about elective amputations of healthy limbs: on the one hand the principle of autonomy is used to deduce the right for body modifications; on the other hand the autonomy of BIID patients is doubted. Neurological results suggest that BIID is a brain disorder producing a disruption of the body image, for which parallels for stroke patients are known. If BIID were a neuropsychological disturbance, which includes missing insight into the illness and a specific lack of autonomy, then amputations would be contraindicated and must be evaluated as bodily injuries of mentally disordered patients. Instead of only curing the symptom, a causal therapy should be developed to integrate the alien limb into the body image. PMID- 19132622 TI - Whose identity is it anyway? PMID- 19132623 TI - The target of the self and the arrows of volition and self-representation. PMID- 19132624 TI - Being whole after amputation. PMID- 19132625 TI - Autonomy is (largely) irrelevant. PMID- 19132626 TI - Two types of autonomy. PMID- 19132627 TI - The relevance of identity in responding to BIID and the misuse of causal explanation. PMID- 19132628 TI - Body integrity identity disorder (BIID) and the limits of autonomy. PMID- 19132630 TI - Are attractive facial characteristics peculiar to the sex of a face? AB - Preferences for faces are thought to be the result of either general adaptations for mate selection, and thus influenced by sexual dimorphism, or mechanisms of general information processing and thus nonspecific to faces. If mate choice determines face preference then it should follow that the sex of a face should affect attractiveness judgements. To test this idea we used image morphing to generate three versions of face images: original, opposite sex, and antiface. First we established that the sex of the face was identifiable in our images. We then collected attractiveness ratings for the three face types. We found that attractiveness ratings to the original faces were correlated with, and did not differ significantly between, ratings to the opposite-sex faces. However, ratings for either the original or opposite face types were not correlated with and were significantly lower than ratings to the antifaces. Our findings failed to support the idea that attractiveness is related to sexual dimorphism in faces alone but suggest instead that other more generic factors influence preferences for all faces. PMID- 19132631 TI - Social gaze cueing to auditory locations. AB - Spatial attention is oriented by social visual cues: targets appearing at cued (gazed-at) locations are detected more rapidly than those appearing at uncued locations. The current studies provide evidence that social gaze directs attention to auditory as well as visual targets at cued locations. For auditory target detection the effect lasted from 300 to 1,005 ms while for discrimination the effect was restricted to 600 ms. Improved performance at 600-ms stimulus onset asynchrony was observed across all experiments and may reflect an optimal processing window for social stimuli. In addition, the orienting of attention by gaze was impaired by the presentation of negative faces. These experiments further demonstrate the unique and cross-modal nature of social gaze cueing. PMID- 19132632 TI - Is advance reconfiguration in voluntary task switching affected by the design employed? AB - In task switching, when the amount of preparation time is increased, a reduction in switch cost or RISC effect is observed. This RISC effect is frequently attributed to advance reconfiguration processes. In the explicit task-cueing procedure, RISC effects are observed when varying the preparation time within participants but not when varying the preparation time across participants--a finding suggesting that RISC effects in the explicit task-cueing procedure are restricted to specific designs. The present study investigated RISC effects in voluntary task switching and compared RISC effects in a within-subjects design with RISC effects in a between-subjects design. Our results indicate that RISC effects are present in both designs. We conclude that advance reconfiguration in voluntary task switching is robust. PMID- 19132633 TI - Attentional orientation induced by temporarily established self-referential cues. AB - Self-referential stimuli such as self-face surpass other-referential stimuli in capture of attention, which has been attributed to attractive perceptual features of self-referential stimuli. We investigated whether temporarily established self referential stimuli are different from other-referential cues in guiding voluntary visual attention. Temporarily established self-referential or friend referential shapes served as central cues in Posner's endogenous cueing task. We found that, relative to friend-referential cues, self-referential cues induced smaller cueing effect (i.e., the difference in reaction times to targets at cued and uncued locations) when the interstimulus interval was short but larger cueing effect when the interstimulus interval was long. Our findings suggest that temporarily established self-referential cues are more efficient to capture reflexive attention at the early stage of perceptual processing and to shift voluntary attention at the later stage of perceptual processing. PMID- 19132634 TI - Effects of frequency allocation on lexical tone identification by Mandarin speaking children with a cochlear implant. AB - CONCLUSION: Frequency allocation with extended frequency ranges yielded significantly higher accuracy in pediatric CI recipients' lexical tone identification. These findings suggest that frequency allocation with extended frequency ranges may be useful in improving lexical tone recognition for at least some pediatric CI recipients. OBJECTIVES: To assess the effects of frequency allocation on lexical tone identification by Mandarin-speaking children with a cochlear implant (CI). SUBJECTS AND METHODS: In a prospective study, 15 prelingually deafened children between 7.17 and 16.17 years of age served as participants. Using Med-el CI devices, each participant's accuracy in lexical tone identification was compared in two conditions: first, the experimental condition, i.e. use of the extended frequency range from 233 to 8501 Hz; second, the control condition, i.e. use of the participant's clinically assigned frequency range from 300 to 8404 Hz. RESULTS: The group mean of pediatric CI users' accuracy in lexical tone identification was 88.02% (SD = 6.31%) in the experimental condition and 83.82% (SD = 9.84%) in the control condition. The group mean was 4.20% (SD = 5.48%) higher in the experimental condition than that in the control condition; this difference was statistically significant (t(14) = 2.97, p=0.010). PMID- 19132635 TI - Management of otitis media-related diseases in children with a cochlear implant. AB - CONCLUSION: In all, 4 of 19 children with a cochlear implant (CI) with otitis media developed cholesteatoma. The insertion of a CI may interfere with normal mastoid pneumatization, stimulate mucosa as foreign body, or act as a nidus of infection. Regular follow-up is necessary for children with a CI at the prevalent ages of otitis media. Antibiotic treatment is suggested whenever acute otitis media is suspected, to prevent chronic progress of infection, cholesteatoma, or even meningitis. OBJECTIVE: To evaluate treatment modality and outcomes of otitis media-related diseases in children with a CI. PATIENTS AND METHODS: This was a retrospective case review in a tertiary referral center of Taipei Medical University, Chi Mei Medical Center. All patients had a history of otitis media or related disease in the implanted ear. Treatments included antibiotics, tympanotomy, and tympanomastoidectomy. RESULTS: In all, 19 of 186 children with a CI were identified as having otitis media, and 4 of them as having cholesteatoma. Among the others, 10 were identified as having acute otitis media, 4 as having middle ear effusion, and 1 as having mastoiditis in the implanted ear. Surgery was performed in children with cholesteatoma and mastoiditis. The CIs of three children were eventually explanted to eradicate cholesteatoma. PMID- 19132636 TI - Polymerized placenta hemoglobin improves cardiac functional recovery and reduces infarction size of isolated rat heart. AB - OBJECTIVE: To investigate the cardioprotective effect of polymerized human placenta hemoglobin (PolyPHb) to rat heart subjected to 8-hour hypothermic storage and 2-hour normothermic reperfusion. METHODS AND RESULTS: Isolated rat hearts were perfused with Langendorff model; after 30 minutes of baseline, the hearts were arrested and stored by St. Thomas' solution (STS) without (STS group) or with 0.5 gHb/dL PolyPHb (PolyPHb group) at 4 degrees C for 8 hours, then reperfused for 2 hours. Compared with STS group, PolyPHb in STS greatly improved the recovery of left ventricular developed pressure (LVDP), maximum LVDP increase and decrease rate (+/-dp/dt), coronary flow rate (CF). Also, both the cardiac enzyme release, including creatine kinase (CK) and lactate dehydrogenase (LDH), and myocardial infarction size were significantly reduced in PolyPHb group. CONCLUSION: Our study demonstrated that the PolyPHb was beneficial to improving cardiac functional recovery and reducing myocardial infarction of 8-hour hypothermic stored rat heart. PMID- 19132637 TI - In-vitro myoglobin clearance by a novel sorbent system. AB - Rhabdomyolysis may lead to acute kidney injury following deposition of myoglobin in renal tubules. Although high-flux dialysis membranes may remove a substantial amount of myoglobin from plasma, this may still not be sufficient to prevent renal damage. We tested a new polymer sorbent, X-Sorb, in vitro to determine its potential to clear myoglobin from solutions. Normal saline or human serum in which myoglobin was dissolved was perfused by a peristaltic pump through a column packed with the sorbent. After a 4-hour perfusion, the myoglobin level in normal saline fell from 200,000 ng/ml to virtually undetectable (<780 ng/ml). Perfusion through the sorbent was then found to lower concentrations of dissolved myoglobin in 3 different 110-ml samples of human serum consistently by > 90% over 4 hours. X-Sorb appears to be an effective sorbent for myoglobin and warrants a trial in vivo to determine whether it is equally effective and safe. PMID- 19132638 TI - Medical application of microbial biopolyesters polyhydroxyalkanoates. AB - Polyhydroxyalkanoates (PHA) are a family of polyesters synthesized by microorganisms under unbalanced growth conditions. PHA including poly-3 hydroxybutyrate (PHB), copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV), poly-4-hydroxybutyrate (P4HB), copolymers of 3-hydroxybutyrate and 3 hydroxyhexanoate (PHBHHx), and poly-3-hydroxyoctanoate (PHO) are now available in sufficient quantity for tissue engineering medical application studies due to their reasonable biocompatibility, adjustable mechanical properties, and controllable biodegradability. This paper reviews many achievements based on PHA for medical devices development, tissue repair, artificial organ construction, drug delivery, and nutritional/therapeutic uses. Combined with the recent FDA approval for P4HB clinical application, one can expect a good prospect for PHA application in the medical fields. PMID- 19132639 TI - Hemoglobin conjugated with a Band 3 N-terminus derived peptide as an oxygen carrier. AB - A peptide composed of 9 amino acids, 7 residues from N-terminus of human erythrocytic Band 3 protein (AcMEELQDD) followed by cysteine and glutamic acids, was conjugated to hemoglobin (Hb) serving as an allosteric effector for oxygen release. The activated polyethylene glycol (PEG), maleimide-PEG-N hydroxysuccinimidyl, was used to crosslink Hb with the peptide. The putative conjugation site on Hb for effective enhancement of oxygen release was characterized as Lys-beta95 by liquid chromatography-tandem mass spectrometry. In addition, the conjugated peptide causes a rightward shift of the oxygen dissociation curve as compared to that of its parent Hb when the degree of oxygen saturation is higher than 50%. Furthermore, this conjugated peptide remains effective on lowering Hb's oxygen affinity after Hb polymerization by another PEG crosslinker. The allosteric properties of the peptide-conjugated Hb may provide a new aspect of Hb-based oxygen carriers. PMID- 19132640 TI - Next steps for clinicians in religious and spiritual therapy: an endpiece. AB - To conclude this issue of the Journal of Clinical Psychology: In Session, the authors identify several next steps for clinicians interested in religious and spiritual therapy. They call for more clinically useful definitions of religion and spirituality and suggest that new methods of clinical practice that employ both Western and Eastern religion and spirituality need to be developed and tested. The need for more clinically focused religious and spiritual assessments is highlighted. They recommend greater clergy-psychotherapist collaboration and propose that clinicians lead and collaborate with researchers to further meaningful research on religion and spirituality. Finally, the authors identify areas of graduate training that require strengthening and provide corresponding guidelines. PMID- 19132641 TI - Clinical psychology training in sleep and sleep disorders. AB - There is growing evidence to suggest that clinical psychologists would benefit from more training in sleep and sleep disorders. Sleep disturbances are commonly comorbid with mental health disorders and this relationship is often bidirectional. In addition, psychologists have become integral members of multidisciplinary sleep medicine teams and there are not enough qualified psychologists to meet the clinical demand. The purpose of this study was to evaluate the current education on sleep and sleep disorders provided to clinical psychology predoctoral students and interns. Directors of graduate programs and internships (N=212) completed a brief online survey on sleep education in their program. Only 6% of programs offers formal didactic courses in sleep, with 31% of programs offering training in the treatment of sleep disorders. There are few programs with sleep faculty (16%), and most reported that their institutions were ineffective in providing sleep education. Thirty-nine percent of training directors reported they would implement a standard curriculum on sleep, if available. The findings from this study suggest that more opportunities are needed for trainees in clinical psychology to gain didactic and clinical experience with sleep and sleep disorders. PMID- 19132642 TI - Intellectual activity and likelihood of subsequently improving or maintaining instrumental activities of daily living functioning in community-dwelling older Japanese: a longitudinal study. AB - OBJECTIVE: To test the a priori hypothesis that increase or maintenance of the level of cognitive activity would be independently associated with a subsequent beneficial impact (i.e. improvement or maintenance) on instrumental activity of daily living (IADL) functioning. METHODS: Observational study using 4-year longitudinal data from 1477 community-dwelling Japanese who were 66 years and older and living in Yoita town, a rural community located in the northwest along the Sea of Japan. Baseline assessment occurred in November 2000. Intellectual Activity and IADL were evaluated using the Tokyo Metropolitan Institute of Gerontology (TMIG) Index of Competence. RESULTS: Increase or maintenance of exposure to intellectual activities over a period of 2 years was associated with increased likelihood of better IADL functioning trajectory (i.e. improvement or stability) over the subsequent 2 years (OR: 1.63; 95% CI: 1.03-2.56; p = 0.035), even after adjustment for major confounders. CONCLUSION: This study documented a longitudinal, independent association of exposure to intellectual activities with better IADL functioning over time. These findings provide preliminary support for promotion of exposure to intellectual activities among older adults as an opportunity to prevent disability in the older segment of the population. Assessment of public health programs targeting such an opportunity is warranted. PMID- 19132643 TI - Is the geriatric depression scale a reliable screening tool for depressive symptoms in elderly patients with cognitive impairment? AB - OBJECTIVE: To determine the reliability of the 30-item Geriatric Depression Scale (GDS-30) for the screening of depressive symptoms in dementia and mild cognitive impairment (MCI) using the Cornell Scale for Depression in Dementia (CSDD) as the 'gold standard'. METHODS: Diagnosed according to strictly applied clinical diagnostic criteria, patients with MCI (n = 156) and probable Alzheimer's disease (AD) (n = 247) were included. GDS-30, CSDD, Mini Mental State Examination (MMSE) and Global Deterioration Scale were assessed in all patients at inclusion. The AD group was divided in three subgroups: mild AD (MMSE>or=18) (n = 117), moderate AD (MMSE< 18 and >or=10) (n = 89) and severe AD (MMSE<10) (n = 38). RESULTS: In MCI patients, moderate but highly significant correlations were found between GDS-30 and CSDD scores (Pearson: r = 0.565; p < 0.001). In mildly (r = 0.294; p = 0.001), moderately (r = 0.273; p = 0.010) and severely (r = 0.348; p = 0.032) affected AD patients, only weak correlations between GDS-30 and CSDD scores were calculated. ROC curve analysis showed that sensitivity and specificity values of respectively 95% and 67% were achieved when a GDS-30 cut-off score of 8 was applied in MCI patients. In AD patients, too low sensitivity and specificity values did not allow selecting an optimal cut-off score by means of ROC curve analysis. CONCLUSION: Using the CSDD as 'gold standard', we demonstrated that the GDS-30 is a reliable screening tool for depressive symptoms in MCI but not in AD patients. PMID- 19132644 TI - Light in aminocatalysis: the asymmetric intermolecular alpha-alkylation of aldehydes. AB - Following the light: Photoredox catalysis along with aminocatalysis have proved to be the right combination for one of the most challenging asymmetric transformation in organic synthesis: the direct intermolecular alpha-alkylation of aldehydes. PMID- 19132645 TI - Nucleation: what happens at the initial stage? AB - Crystallizing growth: The initial structure of crystal nuclei is supersaturation dependent. At low degrees of supersaturation, liquid-like nuclei are formed initially, which undergo a continuous structure transition from liquid-like to crystal-like as the size N increases. This gradual structure evolution substantially lowers the nucleation barrier DeltaG* and facilitates the nucleation relative to the formation of crystal-like clusters from the beginning. PMID- 19132648 TI - [ZAK(R)--Approved drugs for children. Database of children's medicines]. PMID- 19132649 TI - [The in silico Child. Can computer simulations replace clinical pharmacokinetic studies?]. PMID- 19132650 TI - [Ethical aspects of conducting clinical studies in children. An instrumentation loser?]. PMID- 19132651 TI - [Nobel viruses]. PMID- 19132652 TI - [Medicines used for and with children. No, I don't need my drops!]]. PMID- 19132653 TI - [Is green so green]. PMID- 19132654 TI - [Phytotherapy research. A thyme-ivy combination using synergy effects in action and research]. PMID- 19132657 TI - [siRNAs--your friend and helper?]. PMID- 19132669 TI - [Children's medical treatment in the early modern period. "Pro infantibus"]. PMID- 19132671 TI - [Particularities of pharmacotherapy in children. Is there a metamorphosis of "therapeutic orphans"?]. PMID- 19132674 TI - [Pharmazie in unserer Zeit 1/2009]. PMID- 19132675 TI - [Dosage forms for children. After the EU Reform]. PMID- 19132676 TI - [Dispensing and quality control. Treatment options for pediatric patients]. PMID- 19132678 TI - [Regulatory aspects of the drug authorization for children. USA, Europe and Germany]. PMID- 19132679 TI - [Herbal medicines for children. Fennel tea and Co]. PMID- 19132680 TI - Differential protein expression in heart in UT-B null mice with cardiac conduction defects. AB - Cardiac conduction defects were found in transgenic mice deficient in urea transporter UT-B. To investigate the molecular mechanisms of the conduction defects caused by UT-B deletion, we studied the protein expression profiles of heart tissue (comprising most conduction system) in wild-type versus UT-B null mice at different ages. By two-dimensional electrophoresis-based comparative analysis, we found that more than dozen proteins were modulated (>two-fold) in the myocardium of UT-B null mice. Out of these modulated proteins, troponin T (TNNT2) presented significant changes in UT-B null mice at early stage prior to the development of P-R interval elongation, while the change of atrial natriuretic peptide (ANP) occurred only at late stage in UT-B null mice that had the AV block. These data indicate that UT-B deletion caused the dynamic expression regulation of TNNT2 and ANP, and these proteins may provide new clues to investigate the molecular events involved in cardiac conduction. PMID- 19132681 TI - A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates. AB - While protein interaction studies and protein network modeling come to the forefront, the isolation and identification of protein complexes in a cellular context remains a major challenge for plant science. To this end, a nondenaturing extraction procedure was optimized for plant whole cell matrices and the combined use of gel filtration and BN-PAGE for the separation of protein complexes was studied. Hyphenation to denaturing electrophoresis and mass spectrometric analysis allows for the simultaneous identification of multiple (previously unidentified) protein interactions in single samples. The reliability and efficacy of the technique was confirmed (i) by the identification of well-studied plant protein complexes, (ii) by the presence of nonplant interologs for several of the novel complexes (iii) by presenting physical evidence of previously hypothetical plant protein interactions and (iv) by the confirmation of found interactions using co-IP. Furthermore practical issues concerning the use of this 2-D BN/SDS-PAGE display method for the analysis of protein-protein interactions are discussed. PMID- 19132683 TI - Chitosan treatment induces changes of protein expression profile and stilbene distribution in Vitis vinifera cell suspensions. AB - Polyphenols, including stilbenes and flavonoids, are an essential part of human diet and constitute one of the most abundant and ubiquitous groups of plant secondary metabolites, and their level is inducible by stress, fungal attack or biotic and abiotic elicitors. Proteomic analysis of Vitis vinifera (L.) cultivar (cv.) Barbera grape cell suspensions, showed that the amount of 73 proteins consistently changed in 50 microg/mL chitosan-treated samples compared with controls, or between the two controls, of which 56 were identified by MS analyses. In particular, de-novo synthesis and/or accumulation of stilbene synthase proteins were promoted by chitosan which also stimulated trans resveratrol endogenous accumulation and decreased its release into the culture medium. No influence was shown on cis-resveratrol. There was no effect on the accumulation of total resveratrol mono-glucosides (trans- and cis-piceid and trans- and cis-resveratroloside). Throughout the observation period the upregulation of phenylalanine ammonia lyase, chalcone synthase, chalcone flavanone isomerase (CHI) transcript expression levels well correlated with CHI protein amount and with the accumulation of anthocyanins. Chitosan treatment strongly increased the expression of eleven proteins of the pathogenesis related protein-10 family, as well as their mRNA levels. PMID- 19132682 TI - Mass spectrometric and computational analysis of cytokine-induced alterations in the astrocyte secretome. AB - The roles of astrocytes in the CNS have been expanding beyond the long held view of providing passive, supportive functions. Recent evidence has identified roles in neuronal development, extracellular matrix maintenance, and response to inflammatory challenges. Therefore, insights into astrocyte secretion are critically important for understanding physiological responses and pathological mechanisms in CNS diseases. Primary astrocyte cultures were treated with inflammatory cytokines for either a short (1 day) or sustained (7 days) exposure. Increased interleukin-6 secretion, nitric oxide production, cyclooxygenase-2 activation, and nerve growth factor (NGF) secretion confirmed the astrocytic response to cytokine treatment. MS/MS analysis, computational prediction algorithms, and functional classification were used to compare the astrocyte protein secretome from control and cytokine-exposed cultures. In total, 169 secreted proteins were identified, including both classically and nonconventionally secreted proteins that comprised components of the extracellular matrix and enzymes involved in processing of glycoproteins and glycosaminoglycans. Twelve proteins were detected exclusively in the secretome from cytokine-treated astrocytes, including matrix metalloproteinase-3 (MMP-3) and members of the chemokine ligand family. This compilation of secreted proteins provides a framework for identifying factors that influence the biochemical environment of the nervous system, regulate development, construct extracellular matrices, and coordinate the nervous system response to inflammation. PMID- 19132684 TI - Proteomic profiling of antisense-induced exon skipping reveals reversal of pathobiochemical abnormalities in dystrophic mdx diaphragm. AB - The disintegration of the dystrophin-glycoprotein complex represents the initial pathobiochemical insult in Duchenne muscular dystrophy. However, secondary changes in signalling, energy metabolism and ion homeostasis are probably the main factors that eventually cause progressive muscle wasting. Thus, for the proper evaluation of novel therapeutic approaches, it is essential to analyse the reversal of both primary and secondary abnormalities in treated muscles. Antisense oligomer-mediated exon skipping promises functional restoration of the primary deficiency in dystrophin. In this study, an established phosphorodiamidate morpholino oligomer coupled to a cell-penetrating peptide was employed for the specific removal of exon 23 in the mutated mouse dystrophin gene transcript. Using DIGE analysis, we could show the reversal of secondary pathobiochemical abnormalities in the dystrophic diaphragm following exon-23 skipping. In analogy to the restoration of dystrophin, beta-dystroglycan and neuronal nitric oxide synthase, the muscular dystrophy-associated differential expression of calsequestrin, adenylate kinase, aldolase, mitochondrial creatine kinase and cvHsp was reversed in treated muscle fibres. Hence, the re establishment of Dp427 coded by the transcript missing exon 23 has counter-acted dystrophic alterations in Ca2+-handling, nucleotide metabolism, bioenergetic pathways and cellular stress response. This clearly establishes the exon-skipping approach as a realistic treatment strategy for diminishing diverse downstream alterations in dystrophinopathy. PMID- 19132685 TI - Proteomic profiling of tumor cells after induction of telomere dysfunction. AB - Cell division in the absence of telomerase causes progressive telomere shortening which ultimately leads to telomere dysfunction and initiation of genome instability. In order to identify factors related to loss of telomere function, the effects of telomerase inhibition on the proteome of five tumor cell lines were followed by SELDI-TOF-MS. Five differentially expressed protein peaks (p<0.01) were found in a total of 60 clones of five cell lines representing four tissues (lung, breast, prostate, and colon) in which telomerase was inhibited by retroviral overexpression of a dominant negative (DN) mutant of human telomerase reverse transcriptase (hTERT). Among these, a 11.3 kDa peak diminished in DN hTERT clones was identified as histone H4 by nanoflow-HPLC-MS/MS. Immunoblot analysis not only confirmed the decline of histone H4, but also of other core histone proteins including histone H3. Furthermore, upregulation of several cytokeratins was found to be associated with telomere attrition. In conclusion, loss of telomere function is associated with alterations in the proteome which may represent novel biomarkers for the detection of replicative senescence. PMID- 19132686 TI - Comparative proteomics of leaf, stem, and root tissues of synthetic Brassica napus. AB - Comparative proteomics was applied to three vegetative organs of Brassica napus, the leaf, stem, and root using 2-DE. Among the >1600 analyzed spots, 43% were found to be common to all three organs, suggesting the existence of a "basal" or ubiquitous proteome composed of housekeeping proteins. The green organs, leaf, and stem, were closely related (approximately 80% common spots) while the root displayed more organ-specific polypeptides (approximately 10%). Reference maps were established using MS, allowing the identification of 93, 385, and 266 proteins in leaf, stem, and root proteomes, respectively. Bioinformatic analyses were also performed; in silico functional categorization and cellular localization allow obtaining a precise picture of the cell molecular network within vegetative organs. These proteome maps can be explored using the PROTICdb software at the following address: http://bioinformatique.moulon.inra.fr/proticdb/web_view/. PMID- 19132687 TI - Chemical de-O-glycosylation of glycoproteins for application in LC-based proteomics. AB - We describe a cyclic on-column procedure for the sequential degradation of complex O-glycans on proteins or peptides by periodate oxidation of sugars and cleavage of oxidation products by elimination. Desialylated glycoproteins were immobilized to alkali-stable, reversed-phase Poros 20 beads followed by two degradation cycles and the eluted apoproteins were either separated by SDS gel electrophoresis or digested with trypsin prior to LC/ESI-MS. We demonstrate on the peptide and protein level that even complex glycan moieties are removed under mild conditions with only minimal effects on structural integrity of the peptide core by fragmentation, dehydration or by racemization of the Lys/Arg residues. The protocol is applicable on gel-immobilized glycoproteins after SDS gel electrophoresis. Conversion of O-glycoproteins into their corresponding apoproteins should result in facilitated accessibility of tryptic cleavage sites, increase the numbers of peptide fragments, and accordingly enhance protein coverage and identification rates within the subproteome of mucin-type O glycoproteins. PMID- 19132688 TI - Managing the data explosion. A report on the HUPO-PSI Workshop. August 2008, Amsterdam, The Netherlands. AB - The plenary session of the Proteomics Standards Initiative (PSI) of the Human Proteome Organisation at the 7(th) annual HUPO world congress updated the delegates on the current status of the ongoing work of this group. The release of the new MS interchange format, mzML, was formally announced and delegates were also updated on the advances in the area of molecular interactions, protein separations, proteomics informatics and also on PEFF, a common sequence database format currently under review in the PSI documentation process. Community input on this initiative was requested. Finally, the impact these new data standards are having on the data submission process, which increasingly is an integral part of the publication process, was reviewed and discussed. PMID- 19132689 TI - HUPO World Congress Publication Committee meeting. August 2008, Amsterdam, The Netherlands. AB - The plenary session of the Publications Committee of the Human Proteome Organisation at the 7(th) annual HUPO world congress examined the relationship between journals, proteomics standardization initiatives, such as the work of the HUPO-PSI, and the public domain data repositories. Delegates from industry, academia and the publishing houses discussed how best to bring these bodies closer to together and facilitate the publication process for the bench scientist. PMID- 19132690 TI - Norms for the Mini-Mental State Examination from a sample of Sri Lankan older people. AB - OBJECTIVE: To derive norms for the Mini Mental State Examination (MMSE) based on age, gender and level of formal education among the elderly in Sri Lanka. METHOD: A validated Sinhalese version of the MMSE was administered to people aged 55 years and above residing in 14 randomly selected elders' homes. Effects of age, gender and level of formal education on MMSE scores were compared using multiple linear regression analysis. RESULTS: A total of 446 subjects [male/female = 136/320] formed the final sample. Level of education and gender exerted a significant influence on MMSE scores, but not age. The median and 10th percentile scores on the Sinhalese MMSE for the education groups were as follows: education <5 years = 19 and 12; education >5 years = 24 and 16, respectively. CONCLUSION: The findings confirm the influence of level of education on MMSE scores among the elderly living in care homes in Sri Lanka, and suggest that education stratified cut-off scores should be used while screening for cognitive impairment in this population. PMID- 19132691 TI - Cognitive deficits in middle-aged and older adults with bipolar disorder and cognitive complaints: comparison with mild cognitive impairment. AB - OBJECTIVE: Cognitive impairment has been reported in elderly bipolar disorder (BD) patients, however, few studies have evaluated middle-aged and older BD patients using standardized cognitive assessments and none (to our knowledge) analysed middle-aged and older BD patients with recent cognitive complaints. The main objective of this study is to characterize the cognitive deficits of middle aged and older patients with BD and compare them with the common age-related cognitive deficits observed in Mild Cognitive Impairment (MCI). METHODS: For this retrospective study, a systematic search for all cases of BD patients submitted to a neuropsychological assessment from 1999-2007, at participant institutions, was performed, and cases were matched (1:2) by gender and age to a sample of MCI subjects. RESULTS: A total sample of 135 patients, 45 patients with the diagnosis of BD, clinically stable, mean age of 63.8 +/- 8.8 years, and 90 patients with the diagnosis of MCI, mean age of 64.2 +/- 8.4 years, was studied. Patients with MCI were more impaired in verbal memory, whereas BD patients showed more deficits in attention, motor initiative, calculation and verbal abstraction. Interestingly, discriminant analysis classified about half of the BD group as belonging to the MCI group. This BD subgroup showed deficits in episodic memory similar to MCI patients. CONCLUSIONS: Patients with BD and patients with MCI have distinct profiles of cognitive impairment. A subgroup of BD patients with recent cognitive complaints may actually suffer from concomitant incipient MCI, and this finding may have diagnostic and therapeutical implications. PMID- 19132692 TI - Design and synthesis of new trehalose-conjugated pentapeptides as inhibitors of Abeta(1-42) fibrillogenesis and toxicity. AB - Aggregation of the amyloid Abeta peptide and its accumulation into insoluble deposits (plaques) are believed to be the main cause of neuronal dysfunction associated with Alzheimer's disease (AD); small molecules that can interfere with the Abeta amyloid fibril formation are therefore of interest for a potential therapeutic strategy. Three new trehalose-conjugated peptides of the well known beta-sheet breaker peptide iAbeta5p, were synthesized. The disaccharide was covalently attached to different sites of the LPFFD peptide chain, i.e. at the N terminus, C-terminus or at the Asp side chain. CD spectroscopy in different solvents was used to assess changes in the peptide conformation of these compounds. The effects of these glycopeptides on the self-assembly and morphology of Abeta aggregates were investigated by ThT fluorescence assay and dynamic Scanning Force Microscopy, respectively. All the synthesized compounds were tested as inhibitors of Abeta toxicity toward pure cultures of rat cortical neurons. PMID- 19132693 TI - Variants in nicotinic acetylcholine receptors alpha5 and alpha3 increase risks to nicotine dependence. AB - Nicotinic acetylcholine receptors bind to nicotine and initiate the physiological and pharmacological responses to tobacco smoking. In this report, we studied the association of alpha5 and alpha3 subunits with nicotine dependence and with the symptoms of alcohol and cannabis abuse and dependence in two independent epidemiological samples (n = 815 and 1,121, respectively). In this study, seven single nucleotide polymorphisms were genotyped in the CHRNA5 and CHRNA3 genes. In both samples, we found that the same alleles of rs16969968 (P = 0.0068 and 0.0028) and rs1051730 (P = 0.0237 and 0.0039) were significantly associated with the scores of Fagerstrom test for nicotine dependence (FTND). In the analyses of the symptoms of abuse/dependence of alcohol and cannabis, we found that rs16969968 and rs1051730 were significantly associated with the symptoms of alcohol abuse or dependence (P = 0.0072 and 0.0057) in the combined sample, but the associated alleles were the opposite of that of FTND. No association with cannabis abuse/dependence was found. These results suggested that the alpha5 and alpha3 subunits play a significant role in both nicotine dependence and alcohol abuse/dependence. However, the opposite effects with nicotine dependence and alcohol abuse/dependence were puzzling and future studies are necessary to resolve this issue. PMID- 19132694 TI - Reversible control of exo- and endo-budding transitions in a photosensitive lipid membrane. AB - We have developed a method for the photomanipulation of lipid membrane morphology in which the shape of a vesicle can be switched by light through the use of a synthetic photosensitive amphiphile containing an azobenzene unit (KAON12). We prepared cell-sized liposomes from KAON12 and 1,2-dioleoyl-sn-glycero-3 phosphocholine (DOPC) and conducted real-time observations of vesicular transformation in the photosensitive liposome by phase-contrast microscopy. Budding transitions-either budding toward the centre of the liposome (endo-bud) or budding out of the liposome (exo-bud)-could be controlled by light. We discuss the mechanism of this transformation in terms of the change in the effective membrane surface area due to photoisomerization of the constituent molecules. PMID- 19132695 TI - A supported copper hydroxide as an efficient, ligand-free, and heterogeneous precatalyst for 1,3-dipolar cycloadditions of organic azides to terminal alkynes. PMID- 19132696 TI - Tetraalkylammonium salts of weakly coordinating aluminates: ionic liquids, materials for electrochemical applications and useful compounds for anion investigation. AB - In this study, we investigated the tetraalkylammonium salts of the weakly coordinating fluorinated alkoxyaluminates [pftb](-) ([Al(O(C(CF(3))(3))(4)](-)), [hfip](-) ([Al(OC(H)(CF(3))(2))(4)](-)) and [hftb](-) ([Al(OC(CH(3))(CF(3))(2))(4)](-)) in order to obtain information on their undisturbed spectral and structural properties, as well as to study their electrochemical behavior (i.e., conductivities in non-polar solvents and electrochemical windows). Several of the compounds qualify as ionic liquids with melting points as low as 42 degrees C for [NBu(4)](+)[hfip](-). Simple and almost quantitative metathesis reactions yielding these materials in high purity were developed. These [NR(4)](+) salts serve as model compounds for undisturbed anions and their vibrational spectra--together with simulated spectra based on quantum chemical DFT calculations--were used for the clear assignment of the anion bands. Besides, the ion volumes of the anions (V(ion)([pftb](-)) = 0.736 nm(3), V(ion)([hftb](-)) = 0.658 nm(3), V(ion)([hfip](-)) = 0.577 nm(3)) and their decomposition pathways in the mass spectrometric measurements have been established. The salts are highly soluble in non-polar solvents (up to 1.09 mol L(-1) are possible for [NBu(4)](+)[hftb](-) in CH(2)Cl(2) and 0.41 mol L(-1) for [NBu(4)](+)[hfip](-) in CHCl(3)) and show higher molar conductivities if compared to [NBu(4)](+)[PF(6)](-). The electrochemical windows of CH(2)Cl(2), CH(3)CN and 1,2-F(2)C(6)H(4) using the [NBu(4)](+) aluminate electrolytes are up to +0.5 V/ 0.7 V larger than those using the standard [NBu(4)](+)[PF(6)](-). PMID- 19132697 TI - Morphological transformation of Co(OH)2 microspheres from solid to flowerlike hollow core-shell structures. AB - We report, for the first time, a detailed investigation into the formation of highly uniform, 3D, flowerlike, hollow, spherical architectures of cobalt hydroxide through a facile solvothermal process. Various controlling parameters were examined, such as water content in starting materials, reaction time, cobalt(II) precursor concentration, and reaction temperature. On the basis of the experimental results, the formation mechanism of these flowerlike cobalt hydroxide hollow spheres involves aggregation of cobalt hydroxide building clusters into solid spheres and hollowing effect through subsequent dissolution, diffusion, and re-deposition of the smaller crystallites under the surface layer driven by an Ostwald ripening process. Metallic cobalt hollow spheres have also been obtained by thermal decomposition of cobalt hydroxide flowers in a mixed gas of Ar+4 % H(2) at 400 degrees C. The morphology and composition of the products were characterized by X-ray diffraction, field-emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, and (high resolution) transmission electron microscopy. PMID- 19132698 TI - Oxidation of ethers, alcohols, and unfunctionalized hydrocarbons by the methyltrioxorhenium/H2O2 system: a computational study on catalytic C-H bond activation. AB - The potential-energy surfaces (PESs) of methyltrioxorhenium (MTO)-catalyzed C-H insertion reactions in the presence of hydrogen peroxide were studied by accurate DFT methods for a series of substrates including unsaturated hydrocarbons, an ether, and an alcohol. Based on the comprehensive analysis of transition states and intrinsic reaction coordinate (IRC) scans, C-H insertion was found to proceed by a concerted mechanism that does not require, as previously thought, a side-on or a butterfly-like transition state. We found that a typical transition state follows requirements of the S(N)2 reaction instead. Furthermore, by exploring the PESs of several C-H insertion reactions, we discovered that no ionic intermediate is formed even in a polar solvent. The latter was modeled within the self consistent reaction field approach in a polarizable continuum model (PB SCRF/PCM). According to our study, C-H insertion occurs by a concerted but highly asynchronous mechanism that first proceeds by hydride transfer and then turns into hydroxide transfer/rebound. For the oxidation of alcohols, C-H bond cleavage occurs without formation of alkoxide intermediates on the dominant pathway. The computed deuterium kinetic isotope effect of 2.9 for the hydride-transfer transition state for alcohol oxidation is in good agreement with the experimental k(H)/k(D) ration of 3.2 reported by Zauche and Espenson. As confirmed by IRC and PES scans in different solvents, the OH-rebound phase of the C-H insertion pathway demonstrates strong similarities with the rebound mechanism that was previously proposed for cytochrome P450 and metalloporphyrin-catalyzed oxidations. PMID- 19132699 TI - Matrix isolation and spectroscopic characterization of the phenylperoxy radical and its rearranged products. AB - The phenylperoxy radical 1 has been synthesized by the reaction of the phenyl radical 2 with (3)O(2). Radical 1 could be either generated in the gas phase and subsequently trapped in solid argon at 10 K, or directly synthesized in argon matrices. By reacting 2 as well as its perdeuterated isotopomer [D(5)]-2 with (16)O(2) and with (18)O(2), respectively, the four isotopomers [H(5)]-(16)O(2)-1, [D(5)]-(16)O(2)-1, [H(5)]-(18)O(2)-1, and [D(5)]-(18)O(2)-1 were matrix-isolated and characterized by IR spectroscopy. The experimental IR spectra are in excellent agreement with results from DFT calculations. Irradiation of 1 with visible light produces the 2-oxepinoxy radical 5 in a clean reaction. Subsequent irradiation results in ring-opening and formation of several conformers of ketoketene 6. The radicals 1, 5, and 6 play an important role in the combustion of aromatic hydrocarbons and could now be isolated and spectroscopically characterized for the first time. PMID- 19132700 TI - Aluminium speciation in 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)amide/AlCl3 mixtures. AB - Electrodeposition of aluminium is possible from solutions of AlCl(3) dissolved in the 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)amide (C(4)mpyrNTf(2)) ionic liquid. However, electrodeposition is dependant on the AlCl(3) concentration as it only occurs at concentrations >1.6 mol L(-1). At these relatively high AlCl(3) concentrations the C(4)mpyrNTf(2)/AlCl(3) mixtures exhibit biphasic behaviour. Notably, at 1.6 mol L(-1) AlCl(3), aluminium can only be electrodeposited from the upper phase. Conversely, we found that at 3.3 mol L( 1) aluminium electrodeposition can only occur from the lower phase. The complex chemistry of the C(4)mpyrNTf(2)/AlCl(3) system is described and implications of aluminium speciation in several C(4)mpyrNTf(2)/AlCl(3) mixtures, as deduced from Raman and (27)Al NMR spectroscopic data, are discussed. The (27)Al NMR spectra of the C(4)mpyrNTf(2)/AlCl(3) mixtures revealed the presence of both tetrahedrally and octahedrally coordinated aluminium species. Raman spectroscopy revealed that the level of uncoordinated NTf(2)(-) anions decreased with increasing AlCl(3) concentration. Quantum chemical calculations using density functional and ab initio theory were employed to identify plausible aluminium-containing species and to calculate their vibrational frequencies, which in turn assisted the assignment of the observed Raman bands. The data indicate that the electroactive species involved are likely to be either [AlCl(3)(NTf(2))](-) or [AlCl(2)(NTf(2))(2)](-). PMID- 19132701 TI - Self-organization, optical, and electrical properties of alpha-quinquethiophene dinucleotide conjugates. AB - The synthesis and properties of (5')TA(3')-t5 (8a) and (5')CG(3')-t5 (8b) conjugates, in which the self-complementary dinucleotides TA and CG are covalently bound to the central ring of alpha-quinquethiophene (t5), are described. According to molecular mechanics calculations, the preferred conformation of both 8a and 8b is that with the dinucleotide folded over the planar t5 backbone, with the nucleobases facing t5 at stacking distance. The calculations show that the aggregation process of 8a and 8b is driven by a mix of nucleobase-thiophene interactions, hydrogen bonding between nucleobases (non Watson-Crick (W&C) in TA, and W&C in CG), van der Waals, and electrostatic interactions. While 8b is scarcely soluble in any solvents, 8a is soluble in water, indicating that the aggregates of the former are more stable than those of the latter. Microfluidic-induced self-assembly studies of 8a showed the formation of lamellar, spherulitic, and dendritic supramolecular structures, depending on the concentration and solvent evaporation time. The self-assembled structures displayed micrometer dimensions in the xy plane of the substrate and nanometer dimensions in the z direction. Spatially resolved confocal microscopy and spectroscopy showed that the aggregates were characterized by intense fluorescence emission. Cast films of 8a from water solutions showed chirality transfer from the dinucleotide to t5. The hole mobility of the cast films of 8a was estimated using a two-electrode device under high vacuum and found to be up to two orders of magnitude greater than those previously measured for dinucleotide-quarterthiophene conjugates under the same experimental conditions. PMID- 19132702 TI - Zr-zeolite beta: a new heterogeneous catalyst system for the highly selective cascade transformation of citral to (+/-)-menthol. AB - The transformation of citral to menthols involves hydrogenation steps as well as cyclisation of the intermediate, citronellal. The ability of Zr-zeolite beta to catalyse the cyclisation with high diastereoselectivity to (+/-)-isopulegol is the critical step in this cascade transformation. Bifunctional catalysts containing nickel or rhodium supported on Zr-zeolite beta gave menthols in yields of 87-89% and an excellent diastereoselectivity of 94% for the desired (+/-) menthol. Dual catalyst systems of Zr-zeolite beta and nano-dispersed Ni on an MCM 41 support were equally effective and have the added advantage that the rates of the acid- and hydrogenation-catalysed steps can be independently varied. By applying a pressure ramp of 0.2-2 MPa, the yield of menthols could be increased to 95%, with 94% diastereoselectivity for (+/-)-menthol. The low initial pressure minimises the rates of competing hydrogenation reactions to byproducts such as citronellol and 3,7-dimethyloctanol. PMID- 19132703 TI - Regioselectivity control in the metal-catalyzed O-C functionalization of gamma allenols, part 1: Experimental study. AB - We describe versatile regiocontrolled metal-catalyzed heterocyclization reactions of gamma-allenol derivatives leading to a variety of fused enantiopure tetrahydrofurans, dihydropyrans, and tetrahydrooxepines. Regioselectivity control in the O-C functionalization of gamma-allenols can be achieved through the choice of catalyst: use of AuCl(3) exclusively affords tetrahydrofurans, use of La[N(SiMe(3))(2)](3) usually favors the formation of dihydropyrans, whereas use of PdCl(2) solely gives tetrahydrooxepines. In addition, it has been observed that for the Au-catalyzed cycloisomerization, the presence of a methoxymethyl protecting group not only masks a hydroxy functionality, but also exerts directing effects as a controlling unit in a regioselectivity reversal (7-endo versus 5-exo cyclization). In addition, the regioselectivity of the La-catalyzed cycloetherification can be tuned (5-exo versus 7-endo) simply through a subtle variation in the substitution pattern of the allene component (Ph versus Me). Thus, for the first time the regiocontrolled heterocyclization of gamma-allenol derivatives is both catalyst- and substrate-directable. These metal-catalyzed heterocyclization reactions have been developed experimentally (Part 1, this paper), and their mechanisms have additionally been investigated by a theoretical study (Part 2, accompanying paper). PMID- 19132704 TI - Regioselectivity control in the metal-catalyzed functionalization of gamma allenols, part 2: Theoretical study. AB - The gold-, palladium- and lanthanum-catalyzed oxycyclization reactions of azetidin-2-one-tethered gamma-allenol derivatives to a variety of fused enantiopure tetrahydrofurans, dihydropyrans, and tetrahydrooxepines have been developed experimentally (Part 1, accompanying paper). The mechanisms of these regiocontrolled metal-catalyzed heterocyclization reactions have now been computationally explored at the DFT level (Part 2). The energies of the reaction intermediates and transition states for different possible pathways have been calculated in various model systems very close to the real system. Additionally, we selected the La[N(SiH(3))(2)](3) complex to simulate the lanthanide amide precatalyst species. The agreement of theoretically predicted and experimentally observed selectivities is very good in all cases. PMID- 19132705 TI - Applications of multicomponent reactions to the synthesis of diverse heterocyclic scaffolds. AB - The sequencing of multicomponent reactions (MCRs) and subsequent cyclization reactions is a powerful stratagem for the rapid synthesis of diverse heterocyclic scaffolds. The optimal MCR is sufficiently flexible that it can be employed to generate adducts bearing a variety of functional groups that may then be selectively paired to enable different cyclization manifolds, thereby leading to a diverse collection of products. The growing interest in diversity-oriented synthesis has led to increased attention to this paradigm for library synthesis, which has inspired many advances in the design and implementation of MCRs for the construction of diverse heterocyclic scaffolds. PMID- 19132706 TI - Room temperature dynamic polymers based on Diels-Alder chemistry. AB - Dynamers based on reversible Diels-Alder chemistry have been obtained and shown to undergo dynamic exchange at room temperature. Their study in solution by small angle neutron scattering indicated the formation of long and highly flexible chains. Polydispersed molecules gave T(g) values below room temperature, permitting the generation of a dynamic elastomer upon introduction of a dynamic cross-linking agent. The use of a system with a low equilibrium constant gives access to materials with interesting self-healing properties. PMID- 19132707 TI - Copper-catalyzed enantioselective beta-boration of acyclic enones. AB - The enantioselective beta-boration of various acyclic enones has been studied by using chiral diphosphine-copper complexes. Good to excellent yields and enantioselectivities (up to 97 % ee (enantiomeric excess)) were observed for a range of substrates under optimized conditions. In this transformation, the addition of a controlled amount of alcohol, especially methyl alcohol, is critical to obtain products in high ee and yield. This methodology accommodates structural variation of acyclic enones and provides access to a range of functionalized chiral organoboronates in high enantiomeric purity. PMID- 19132708 TI - Highly efficient access to bi- and tricyclic ketals through gold-catalyzed tandem reactions of 4-acyl-1,6-diynes. AB - Single step: Fused bicyclic and bridged tricyclic ketals were synthesized in a single step from the reactions of easily available 4-acyl-1,6-diynes with H(2)O and alkanols (see scheme). The highly efficient AuCl(3)-catalyzed multicomponent domino reactions, involving five C-O bond formations, can proceed in a highly regio- and diastereoselective manner at room temperature under air and lead to structures of high molecular complexity from simple starting materials in an atom economic way. PMID- 19132709 TI - Dual organocatalysis: asymmetric allylic-allylic alkylation of alpha,alpha dicyanoalkenes and Morita-Baylis-Hillman carbonates. AB - The first highly enantioselective allylic-allylic alkylation of alpha,alpha dicyanoalkenes and Morita-Baylis-Hillman carbonates by dual catalysis of (DHQD)(2)AQN and (S)-BINOL has been investigated. Excellent stereoselectivities have been achieved for a broad spectrum of substrates (d.r. > 99:1, up to 99 % ee). The multifunctional allylic products could be efficiently converted to a range of complex chiral cyclic frameworks. EWG = electron-withdrawing group, (DHQD)(2)AQN = hydroquinidine (anthraquinone-1,4-diyl) diether, (S)-BINOL = (S)-( )-1,1'-bi-2-naphthol. PMID- 19132710 TI - Evidence that putative ADHD low risk alleles at SNAP25 may increase the risk of schizophrenia. AB - Synaptosomal Associated Protein 25 kDa (SNAP25) has been implicated in the pathogenesis of schizophrenia by numerous neuropathological studies and genetic variation at SNAP25 has been reported to be associated with ADHD. Expression levels of the putative schizophrenia susceptibility gene DTNBP1 has been shown to influence the levels of SNAP25 in vitro. We undertook directed mutation screening of SNAP25 in UK schizophrenic cases followed by direct association analysis of all variants identified and identified known exonic SNPs that showed evidence for association (rs3746544 P = 0.004 OR = 1.26, rs8636 P = 0.003 OR = 1.27), although these SNPs are highly correlated (r(2) > 0.99). We additionally genotyped a further 31 tag SNPs spanning the SNAP25 locus and identified several independent SNPs that were nominally associated with schizophrenia (strongest association at rs3787283, P = 0.006, OR = 1.25) however, due to the number of tests performed no SNP met experiment-wise significance (minimum permuted P-value = 0.1). Post hoc analysis revealed that the SNPs nominally associated with schizophrenia (rs3787283, rs3746544) were the same as those previously demonstrated to be associated with ADHD but with the opposite alleles, allowing the intriguing hypothesis that genetic variation at SNAP25 may be differentially associated with both schizophrenia and ADHD. PMID- 19132711 TI - Does perception of the childbirth experience predict women's early parenting behaviors? AB - Evidence regarding the predictors of positive parenting behaviors in the early transition to parenting is inconsistent and limited. In this prospective, cohort study, we examined whether women's perceptions of their childbirth experience, as well as selected demographic, obstetrical, and psychosocial variables, predicted positive parenting behaviors at 1 month postpartum in 175 Canadian mothers. Women's birth experience did not predict early parenting behaviors, however being better educated and having a vaginal birth did. Excellent partner support and maternal mental health were also significantly associated with positive parenting at 1 month. Nurses have a responsibility to assess women for possible risks for sub-optimal parenting, based on the predictors found, and intervene to enhance parenting behaviors. PMID- 19132712 TI - A non-randomized study to investigate the effects of the atypical antipsychotic aripiprazole on the steady-state pharmacokinetics of lamotrigine in patients with bipolar I disorder. AB - OBJECTIVE: To determine the effect of aripiprazole on steady-state pharmacokinetics of lamotrigine in patients with bipolar I disorder who were clinically stable on lamotrigine (100-400 mg/day) for >or=4 weeks. METHODS: In this open-label study, aripiprazole was administered at 10 mg/day for 3 days, 20 mg/day for 3 days, then 30 mg/day for 8 days. Blood samples were collected on Days -1 and 14 for determination of lamotrigine steady-state pharmacokinetic parameters. Safety and tolerability were assessed. RESULTS: Eighteen patients were administered aripiprazole in combination with lamotrigine. Geometric mean (GM) values for lamotrigine maximum plasma concentration were similar for lamotrigine alone (26 ng/mL) and with co-administered aripiprazole (23 ng/mL). GM values for plasma lamotrigine area under the concentration-time curve (AUCtau) were comparable for lamotrigine alone (434 ng/h/mL) and with co-administered aripiprazole (394 ng/h/mL). Median T(max) of lamotrigine alone and combined with aripiprazole was 1.98 and 0.77 h, respectively. No changes to lamotrigine dose normalized plasma trough concentrations were observed with co-administered aripiprazole. Sixteen patients (88.9%) experienced >or=1 adverse event (AE), the most common of which was insomnia (n = 6). CONCLUSIONS: Aripiprazole had no meaningful effect on lamotrigine steady-state pharmacokinetics in patients with bipolar I disorder. No dosage adjustment of lamotrigine is required and the combination was generally safe and well tolerated. PMID- 19132713 TI - Transition metals as electron traps. I. Structures, energetics, electron capture, and electron-transfer-induced dissociations of ternary copper-peptide complexes in the gas phase. AB - Electron-induced dissociations of gas-phase ternary copper-2,2'-bipyridine complexes of Gly-Gly-Gly and Gly-Gly-Leu were studied on a time scale ranging from 130 ns to several milliseconds using a combination of charge-reversal ((+)CR(-)) and electron-capture-induced dissociation (ECID) measured on a beam instrument and electron capture dissociation (ECD) measured in a Penning trap. Charge-reduced intermediates were observed on the short time scale in the (+)CR( ) and ECID experiments but not in ECD. Ion dissociations following electron transfer or capture mostly occurred by competitive bpy or peptide ligand loss, whereas peptide backbone fragmentations were suppressed in the presence of the ligated metal ion. Extensive electron structure theory calculations using density functional theory and large basis sets provided optimized structures and energies for the precursor ions, charge-reduced intermediates, and dissociation products. The Cu complexes underwent substantial structure changes upon electron capture. Cu was calculated to be pentacoordinated in the most stable singly charged complexes of the [Cu(peptide-H)bpy](+*) type where it carried a approximately +1 atomic charge. Cu coordination in charge-reduced [Cu(peptide-H)bpy] intermediates depended on the spin state. The themodynamically more stable singlet states had tricoordinated Cu, whereas triplet states had a tetracoordinated Cu. Cu was tricoordinated in stable [Cu(peptide-H)bpy](-*) products of electron transfer. [Cu(peptide)bpy](2+*) complexes contained the peptide ligand in a zwitterionic form while Cu was tetracoordinated. Upon electron capture, Cu was tri- or tetracoordinated in the [Cu(peptide)bpy](+) charge-reduced analogs and the peptide ligands underwent prototropic isomerization to canonical forms. The role of excited singlet and triplet electronic states is assessed. PMID- 19132714 TI - Chemical cross-linking with NHS esters: a systematic study on amino acid reactivities. AB - Structure elucidation of tertiary or quaternary protein structures by chemical cross-linking and mass spectrometry (MS) has recently gained importance. To locate the cross-linker modification, dedicated software is applied to analyze the mass or tandem mass spectra (MS/MS). Such software requires information on target amino acids to limit the data analysis time. The most commonly used homobifunctional N-hydroxy succinimide (NHS) esters are often described as reactive exclusively towards primary amines, although side reactions with tyrosine and serine have been reported. Our goal was to systematically study the reactivity of NHS esters and derive some general rules for their attack of nucleophilic amino acid side chains in peptides. We therefore studied the cross linking reactions of synthesized and commercial model peptides with disuccinimidyl suberate (DSS). The first reaction site in all cases was expectedly the alpha-NH(2)-group of the N-terminus or the epsilon-NH(2)-group of lysine. As soon as additional cross-linkers were attached or loops were formed, other amino acids were also involved in the reaction. In addition to the primary amino groups, serine, threonine and tyrosine showed significant reactivity due to the effect of neighboring amino acids by intermediate or permanent Type-1 cross link formation. The reactivity is highly dependent on the pH and on adjacent amino acids. PMID- 19132715 TI - Synovial sarcoma of the Eustachian tube. AB - BACKGROUND: Synovial sarcoma is a rare, aggressive high-grade neoplasm with poor prognosis. Synovial sarcoma is usually encountered in the lower extremities of young adults; only a few cases have been reported in the head and neck region. METHODS: We describe a case of a 44-year-old man seen with a 3-month history of progressive nasal obstruction. Nasoendoscopy revealed a pink polypoid mass filling in the entire right posterior nasal cavity. CT and MRI studies showed that a hyperintense mass on the right nasopharynx extending to the nasal cavity anteriorly and the oropharynx inferiorly. During the operation, the mass was noted as originating from the lower portion of anterior cushion of the right Eustachian tube. We endoscopically removed the tumor and part of the anterior cushion of the right Eustachian tube. Because of the large tumor size (7.0 x 3.5 cm), we took the tumor out transorally. RESULTS: The histopathology and genetic analysis confirmed the diagnosis of monophasic synovial sarcoma prominent myxoid pattern. The patient then underwent adjuvant irradiation to 64.8 Gy, and he was free of recurrence and distant metastasis in the 5-year follow-up. CONCLUSION: To our knowledge, this is the first case report of synovial sarcoma of the Eustachian tube. Synovial sarcoma should be included in the differential diagnosis when managing the Eustachian tube region mass. Recommended treatment is surgery and postoperative adjuvant radiotherapy. PMID- 19132716 TI - Effectiveness of salvage selective and modified radical neck dissection for regional pathologic lymphadenopathy after chemoradiation. AB - BACKGROUND: The objective of this study was to evaluate the effectiveness and safety of our careful observational strategy and neck dissections and the accuracy of ultrasound-guided fine-needle aspiration cytology, and to determine the prognostic factors for outcome and regional control after primary chemoradiation. Diagnostic evaluation of the regional status after concurrent chemoradiation for advanced head and neck cancer remains difficult, and the indications for a salvage neck dissection and its extent are not clearly defined. METHODS: In a series of 540 patients, there was suspicion of regional residual or recurrent disease after chemoradiation in 61 patients who underwent 68 salvage neck dissections and 68 patients who were considered unresectable. For the patients with salvage neck dissection, the accuracy of ultrasound-guided fine needle aspiration cytology was determined. Disease control in the neck, disease specific and overall survival, and parameters that may have prognostic value for the outcome were evaluated. RESULTS: Neck dissection specimens contained viable tumor in 26 (43%) patients. Of these, 13 had selective neck dissections and 13 modified radical neck dissections. Ultrasound-guided fine-needle aspiration cytology had a sensitivity of 80% and specificity of 42%. Nine patients developed a regional recurrence after salvage neck dissection (5 located in contralateral neck). Five-year regional control and overall survival rates were 79% and 36%, respectively. Significant prognostic factors for overall survival were surgical margins and "residual versus recurrent disease" in multivariate analysis. CONCLUSION: Considering the good regional control rate and the high rate of unnecessary neck dissections with a theoretical planned neck dissection strategy, we conclude that a careful observational strategy is worthwhile and safe. For the evaluation of radiation treatment response, ultrasound-guided fine-needle aspiration cytology has a low specificity. PMID- 19132717 TI - Elective neck dissection for the management of the N0 neck in early cancer of the oral tongue: need for a randomized controlled trial. AB - BACKGROUND: The aim of this study is to determine the need for a randomized controlled trial in order to define the role of an elective neck dissection (END) in the treatment of early tongue cancers. METHODS: We present a large retrospective analysis of patients with T1-2 N0 squamous cell cancers of the oral anterior tongue treated at a single institution. A total of 359 eligible patients with early tongue cancers were divided into 2 groups: END and wait and watch (WW). An analysis for survival outcomes and prognostic factors was conducted. RESULTS: The estimated 3- and 5-year disease-free survival for the END group was 76% and 74% versus 71% and 68% for the WW group, respectively (p = .53). The 3- and 5-year overall survival (OS) rate for the END group was 69% and 60% versus 62% and 60% for the WW group, respectively (p = .24). Tumor grade and perineural invasion were independent predictors of recurrence. CONCLUSION: END did not impact disease-free or OS. Current literature still remains divided on this issue emphasizing the need for a randomized controlled trial. PMID- 19132718 TI - Extramammary myofibroblastoma in the head and neck region. AB - BACKGROUND: Mammary myofibroblastoma is a benign mesenchymal tumor of the breast. Extramammary location is very rare and usually occurs along the embryonic milk line. METHODS AND RESULTS: In this article, we present a case of a woman with an infra-auricular swelling. A diagnosis could not be made on CT scans, so the mass was surgically removed. Histological examination showed spindle cells in clusters, with CD34, desmin, and actin expression. The diagnosis of extramammary myofibroblastoma was made. CONCLUSION: We describe this case because of the unusual location of this rare tumor. Furthermore, we want to highlight the possible relationship between (extra)mammary myofibroblastoma and spindle cell lipoma (SCL). SCL has a predilection site in the posterior neck. Both tumors show the same clinical behavior, a close histological resemblance, and their cells are both immunoreactive for CD34 and CD10. The loss of RB/13q14 and FKHR/13q14 loci, which is typically found in SCL cells, was also shown in this case. PMID- 19132719 TI - Radiotherapy for perineural invasion in cutaneous head and neck carcinomas: toward a risk-adapted treatment approach. AB - BACKGROUND: We retrospectively reviewed outcomes in patients treated with radiotherapy (RT) for cutaneous head and neck carcinoma with perineural invasion (PNI), with the aim of developing risk-adapted treatment guidelines. METHODS: A total of 118 patients were treated with RT between April 1992 and July 2000. Ninety-seven patients had PNI discovered through histology (pPNI) and 21 patients had symptoms/signs of PNI (cPNI). All received RT (median dose, 55 Gy; range, 17 74): 114 postoperatively and 4 definitively. Median follow-up was 84 months (range, 4-201). RESULTS: The 5-year local control (LC) rates were 90% with pPNI and 57% with cPNI (p < .0001). The pPNI and cPNI groups also differed in relapse free survival (76% vs 46%, p = .003), disease-specific survival (90% vs 76%, p = .002), and overall survival (69% vs 57%, p = .03). pPNI patients with BCC histology (n = 42) had better LC (97% vs 84%, p = .02) than pPNI SCC (n = 55). CONCLUSION: Surgery plus RT provides a high rate of LC in patients with pPNI, particularly those with BCC. Therapeutic improvements are needed for patients with cPNI. PMID- 19132720 TI - Endoscopic laser surgery of early glottic cancer: involvement of the anterior commissure. AB - BACKGROUND: Early glottic cancer can be cured with transoral laser resection, but in cases with anterior commissure involvement, there is still controversy concerning the best treatment modality. METHODS: The impact of anterior commissure involvement on local control was analyzed in a retrospective review of 444 patients with early glottic cancer (pT1a-pT2a) treated between 1986 and 2004 with transoral laser microsurgical resection. RESULTS: The anterior commissure was involved in 153 cases; the 5-year local control rate with and without anterior commissure involvement was 73% versus 89% for T1a and 68% versus 86% for T1b tumors. For T2a lesions, the 5-year local control rate was 76%, irrespective of anterior commissure involvement. CONCLUSION: In early glottic cancer treated by transoral laser microsurgery, a decrease in local control is evident in case of anterior commissure involvement for T1a and T1b but not for T2a tumors. PMID- 19132721 TI - Alternative reconstructive choices for anterolateral thigh flap dissection in cases in which no sizable skin perforator is available. AB - BACKGROUND: Alternative choices were proposed to facilitate a successful reconstruction when no sizable skin perforator is encountered in anterolateral thigh (ALT) flap dissection. METHODS: Alternative choices such as harvest of a tensor fascia latae (TFL) flap, elevation of an anteromedial thigh (AMT) flap, use of a free muscle component with full-thickness skin graft coverage, and use of another donor flap were performed. RESULTS: Between August 1995 and December 2006, 10 of 923 patients underwent ALT flap elevation with no sizable perforators. Of them, each of the 4 patients received reconstruction with a free TFL flap, 3 with an AMT flap, 2 with a free muscle flap, and 1 with a radial forearm flap. There was 1 TFL flap failure due to postoperative venous thrombosis, and the defect was repaired by an AMT flap. CONCLUSION: When no sizable perforator is encountered, successful reconstruction could still be achieved by the proposed alternative choices. PMID- 19132722 TI - Evaluation of treatment results with regard to initial anterior commissure involvement in early glottic carcinoma treated by external partial surgery or transoral laser microresection. AB - BACKGROUND: Modalities of surgical treatment of early glottic carcinoma include transoral laser microresection and external partial surgery. METHODS: This is a retrospective analysis of 119 glottic carcinomas treated by external partial surgery (57 pT1a, 1 pT1b, 10 pT2) or transoral laser microresection (46 pT1a, 4 pT1b, 1 pT2) with special regard to initial anterior commissure involvement. RESULTS: Local recurrence in external partial surgery was 12%. Three- and 5-year local control was 86%. Local recurrence in transoral laser microresection was 16%. Three- and 5-year local control was 88% and 70%, respectively. No significant correlation was found between local control and surgical approach. An analysis of all 119 tumor revealed that anterior commissure involvement significantly decreased local control. CONCLUSION: Initial anterior commissure involvement was associated with a higher risk of local recurrence. Overall, treatment of glottic carcinoma involving the anterior commissure requires much experience and advanced surgical skills regardless which technique is preferred. PMID- 19132723 TI - Positive personal changes in the aftermath of head and neck cancer diagnosis: a qualitative study in patients and their spouses. AB - BACKGROUND: There is increasing evidence that serious and life-threatening illnesses such as malignant tumors not only can lead to increased psychological stress, but also can lead to positive changes. METHODS: In this qualitative study, 31 patients with head and neck cancer and 25 women partners were interviewed with regard to positive personal changes (posttraumatic growth). The audiotaped responses were analyzed using qualitative context analysis. RESULTS: A total of 25 patients (81%) and their women partners (84%) reported positive changes. Qualitative content analysis revealed 3 different categories of growth: attitudes toward life, personal strength, and relationships. Partners reported significantly more positive changes in relationships, especially, within the partnership. The total amount of positive change was almost equal. CONCLUSION: Positive personal changes are frequently reported by patients and by significant others. The total amount of positive change is almost equal in patients and their women partners, but wives experience more positive changes within the partnership. PMID- 19132724 TI - Expression modes and clinical manifestations of latent membrane protein 1, Ki-67, cyclin-B1, and epidermal growth factor receptor in nonendemic nasopharyngeal carcinoma. AB - BACKGROUND: We aimed to identify clinical significance of latent membrane protein 1 (LMP1), Ki-67, cyclin-B1, and epidermal growth factor receptor (EGFR), in nonendemic nasopharyngeal carcinoma (NPC). METHODS: The relation between expression of the markers in 45 NPC specimens and clinicopathological and survival variables was statistically analyzed. RESULTS: LMP1 was present in 33% of the tumors, and its presence was associated with advanced nodal and disease stages. Overexpression was defined as labeling index > or = median value for Ki 67, > or = 15% for cyclin-B1, and > or =50% for EGFR, and it was displayed in 50%, 55%, and 80% of the specimens, respectively. Strong EGFR staining intensity and not overexpression of the 3 markers was the variable with statistically significant impact on treatment outcomes in terms of worse local and locoregional tumor control rates. CONCLUSIONS: Our results suggest that the evaluation of EGFR staining intensity in patients with NPC may identify a subgroup of patients with poor prognosis. PMID- 19132726 TI - Length change patterns of the extensor retinaculum and the effect of total knee replacement. AB - Patellofemoral dysfunction following total knee replacement (TKR) is a significant clinical problem, but little information exists on the mechanics of the patellofemoral retinacula or the effects of TKR on these structures. We hypothesized that TKR would cause significant elongation of the retinacula. Retinacular length changes were measured by threading sutures along the retinacula, fixing the sutures to the patella and the iliotibial band (ITB), and attaching the femoral ends to displacement transducers. The intact knee was flexed-extended while the quadriceps and ITB were tensed and the retinacular length change patterns were recorded. The measurements were repeated post-TKR. The medial patellofemoral ligament (MPFL) was close to isometric, stretching 2 mm in terminal knee extension, whereas the lateral retinaculum slackened 8 mm from 110 degrees to 0 degrees flexion. TKR did not cause significant elongation of either of the retinacula, the largest change being 3 mm elongation of the MPFL around 40 degrees , which stretched the MPFL by 1.4 mm above its maximum natural length. Thus, this work did not support the hypothesis that TKR causes significant elongation of the retinacula sufficient to affect knee function. PMID- 19132727 TI - Cobalt ions induce chemokine secretion in primary human osteoblasts. AB - Chemokines are major regulators of the inflammatory response and have been shown to play an important role in periprosthetic osteolysis. Titanium particles have previously been shown to induce IL-8 and MCP-1 secretion in osteoblasts. These chemokines result in the chemotaxis and activation of neutrophils and macrophages, respectively. Despite a resurgence in the use of cobalt-chromium molybdenum alloys in metal-on-metal arthroplasty, cobalt and chromium ion toxicity in the periprosthetic area has been insufficiently studied. In this study we investigate the in vitro effect of cobalt ions on primary human osteoblast activity. We demonstrate that cobalt ions rapidly induce the protein secretion of IL-8 and MCP-1 in primary human osteoblasts. This elevated chemokine secretion is preceded by an increase in the transcription of the corresponding chemokine gene. Using a Transwell migration chemotaxis assay we also demonstrate that the chemokines secreted are capable of inducing neutrophil and macrophage migration. Furthermore, cobalt ions significantly inhibit osteoblast function as demonstrated by reduced alkaline phosphatase activity and calcium deposition. In aggregate these data demonstrate that cobalt ions can activate transcription of the chemokine genes IL-8 and MCP-1 in primary human osteoblasts. Cobalt ions are not benign and may play an important role in the pathogenesis of osteolysis by suppressing osteoblast function and stimulating the production and secretion of chemokines that attract inflammatory and osteoclastic cells to the periprosthetic area. PMID- 19132728 TI - Hypofractionated stereotactic radiotherapy for the treatment of brain metastases. AB - BACKGROUND: This retrospective review evaluated the efficacy and toxicity profiles of various dose fractionations using hypofractionated stereotactic radiotherapy (HSRT) in the treatment of brain metastases. METHODS: Between 2004 and 2007, 36 patients with 66 brain metastases were treated with HSRT. Nine of these subjects were excluded because of the absence of post-treatment magnetic resonance imaging scans, resulting in 27 patients with a total of 52 lesions. Of these 52 lesions, 45 lesions were treated with whole-brain radiotherapy plus a HSRT boost and 7 lesions were treated with HSRT as the primary treatment. The median prescribed dose was 25 grays (Gy) (range, 20 Gy-36 Gy) with a median of 5 fractions (range, 4 fractions-6 fractions) to a median 85% isodose line (range, 50%-100%). The median follow-up interval was 6.6 months (range, 0.9 months-26.8 months). RESULTS: The median overall survival time was 10.8 months, and 66.7% of patients died of disease progression. After HSRT treatment of 52 brain lesions, 13 lesions demonstrated complete responses, 12 lesions demonstrated partial responses, 22 lesions demonstrated stable disease, and 5 lesions demonstrated progressive disease. Actuarial local tumor control rates at 6 months and 1 year were 93.9% and 68.2%, respectively. Maximum tumor dimension, concurrent chemotherapy, and a tumor volume <1 cc were found to be statistically significant factors for local tumor control. One patient had a grade 3 toxicity (according to National Cancer Institute Common Terminology Criteria for Adverse Events). CONCLUSIONS: HSRT provides a high level of tumor control with minimal toxicity comparable to single-fraction stereotactic radiosurgery (SRS). The results of the current study warrant a prospective randomized study comparing single-fraction SRS with HSRT in this patient population. PMID- 19132729 TI - Pharmacokinetics of high-dose methotrexate in infants treated for acute lymphoblastic leukemia. AB - BACKGROUND: Interfant-99 was an international collaborative treatment protocol for infants with acute lymphoblastic leukemia (ALL). PROCEDURE: We collected data on 103 infants at the time of their first treatment with high-dose methotrexate (HD MTX), 5 g/m(2). Children <6 months of age received two-third of the calculated dose based on body surface area (BSA), children 6-12 months three fourth of the calculated dose, and children >12 months full dose. RESULTS: The median steady-state MTX concentration at the end of the 24-hr infusion was 57.8 microM (range 9.5-313). The median systemic clearance was 6.22 L/hr/m(2) BSA, and tended to increase with age (P = 0.099). Boys had higher clearance than girls, 6.77 and 5.28 L/hr/m(2) (P = 0.030), and tended to have lower median MTX concentration at 24 hr. Eight infants had MTX levels below 20 microM, a level judged to be sufficient in B-lineage ALL in children >1 year of age. All infants tolerated the dose well enough to receive a second dose of HD MTX without dose reduction. We found no significant effect on disease-free survival for MTX steady state concentration, MTX clearance, or time to MTX below 0.2 microM. CONCLUSIONS: Our data provide no support for a change in the dosing rules for MTX used in Interfant-99. However, in view of the poor treatment results for infants, one might consider increase in the dose for patients who reach plasma levels below median after the first MTX dose. PMID- 19132730 TI - Neural distribution of nonapeptide binding sites in two species of songbird. AB - Vasotocin (VT) and its mammalian homologue, vasopressin (VP), modulate many social behaviors in a variety of vertebrate species. In songbirds, the effects of centrally administered VT vary according to species, which may reflect species specific distributions of VT binding sites. Different radioligands used to map receptors in previous autoradiographical studies have revealed nonoverlapping distributions of VT binding, suggesting a heterogeneous population of more than one type of VT receptor. For two model songbird species, the white-throated sparrow (Zonotrichia albicollis) and zebra finch (Taeniopygia guttata), we labeled putative VT receptors with two radioligands, [(125)I]ornithine vasotocin analog ([(125)I]OVTA) and [(125)I]linear VP antagonist ([(125)I]HO-LVA). Competitive binding assays in the lateral septum showed that both ligands were effectively displaced by both VT and a related nonapeptide, mesotocin (MT), showing that these radioligands, which were developed to label mammalian nonapeptide receptors, label at least one population of related receptors in songbirds. [(125)I]OVTA labeled receptors throughout the telencephalon, diencephalon, midbrain, and brainstem, with a similar distribution in both species. In contrast, the binding of [(125)I]HO-LVA was restricted to the septal area, dorsal arcopallium, and optic tectum in sparrow and was essentially undetectable in zebra finch. Because the avian brain is likely to express multiple types of VT receptors, we hypothesize that the binding patterns of these radioligands represent a heterogeneous receptor population. PMID- 19132731 TI - Dialects in pygmy marmosets? Population variation in call structure. AB - Population variation in primate vocal structure has been rarely observed. Here, we report significant population differences in the structure of two vocalizations in wild pygmy marmosets (Trills and J calls). We studied 14 groups of pygmy marmosets Callithrix (Cebuella) pygmaea pygmaea from five populations in northeastern Ecuador. We analyzed the acoustic structure of Trills and J calls recorded from two adult animals in each group through focal samples. Although individuals and groups within a population differed in call structure, we found consistent structural differences at a population level for Trills and J calls. Pair-wise comparisons for the two call types point to San Pablo and Amazoonico as the populations that differed the most, whereas Hormiga and Zancudococha showed the least differences. Discriminant function analysis indicates that calls from each population could be classified accurately at rates significantly above chance. Habitat acoustics, social factors and genetic drift may explain interpopulation vocal differences. This is the first evidence of within subspecies vocal differences, or dialects, in wild populations of a neotropical primate species. PMID- 19132732 TI - Quantitative variability of cyanogenesis in Cathariostachys madagascariensis-the main food plant of bamboo lemurs in Southeastern Madagascar. AB - Giant bamboo (Cathariostachys madagascariensis) is a major food plant for three sympatric species of bamboo-eating lemurs (Hapalemur aureus, H. griseus, and Prolemur simus) in the rain forests of southeastern Madagascar. This plant species is strongly cyanogenic. However, quantitative data on cyanide concentration in C. madagascariensis are scarce. Previous studies reported 15 mg cyanide per 100 g fresh shoot material (corresponding to approx. 57 micromol cyanide per gram dry weight). However, we found mean concentrations (+/-SE) ranging from 139.3+/-19.32 in ground shoots to 217.7+/-16.80 micromol cyanide per gram dry weight in branch shoots. Thus, cyanogenesis of C. madagascariensis was up to four times higher than reported before. In contrast to the strongly cyanogenic shoots no cyanide could be detected in differently aged leaves of C. madagascariensis confirming earlier studies. Within individual shoots fine-scaled analysis revealed a characteristic ontogenetic pattern of cyanide accumulation. Highest concentrations were found in youngest parts near the apical meristem, whereas concentrations decreased in older shoot parts. Beyond the general intra individual variability of cyanogenic features analyses indicated site-specific variability of both, the ontogenetic pattern of cyanide concentration as well as the total amount of cyanide accumulated in shoots. Additionally, analyses of soluble proteins-one important nutritive measure affecting food plant quality demonstrated a converse quantitative relation of protein concentrations in leaves to cyanide concentration in shoots at the site-specific level. We, thus, suggest integrative analyses on quantitative variation of cyanogenesis together with nutritive plant parameters in future studies. This approach would allow obtaining more detailed insights into spatial variability of giant bamboo's overall browse quality and its impact on lemur herbivores. PMID- 19132733 TI - The KK-Periome database for transcripts of periodontal ligament development. AB - The periodontal ligament (PDL) is a strong connective tissue that surrounds the tooth root, absorbs occlusal forces, and functions as a sense organ. PDL originated from dental follicle (DF), which possessed mesenchymal progenitors in the developing tooth germ. However, as specific marker genes for PDL and DF are currently unavailable, the molecular mechanisms of PDL development are yet to be clarified. To facilitate the identification of such genes, we have previously established a transcriptome database of the human PDL (the KK-Periome database) and screened for specific genes expressed during PDL development. Initial screening of the database revealed two marker genes for distinguishing DF and PDL. The KK-Periome database thus appears to offer a useful resource for investigating genes involved in PDL development. PMID- 19132734 TI - Spermiogenic nuclear protein transitions and chromatin condensation. Proposal for an ancestral model of nuclear spermiogenesis. AB - We have chosen three species (Sparus aurata, Dicentrarchus labrax, and Monodonta turbinata) that represent different transition patterns in the composition and structure of spermiogenic nuclei. The transition patterns of these species are representative of spermiogenesis in a large number of animal species. We analyze: (a) nuclear protein exchange; (b) chromatin condensation pattern; and (c) histone acetylation during spermiogenic development. In the simplest spermiogenesis histones and nucleosomes remain in mature sperm. Chromatin of spermatids is organized into 20 nm granules, simultaneous with a nuclear volume reduction. The granules coalesce in the final stage of spermiogenesis. Granular chromatin is correlated with acetylation of histones H3 and H4, whereas final coalescence is associated with histone deacetylation. We also studied two other spermiogenesis where a basic protein substitutes histones. Each species has a very different substituting protein. One has a typical protamine of 34 amino acids; the other has a sperm nuclear basic proteins (SNBP) of 106 amino acids. In both, the structural transitions and histone acetylation pattern are similar: in early spermiogenesis chromatin is organized into 20 nm granules, and histones are significantly acetylated, while the nuclear volume decreases. Subsequently, acetylated histones are displaced by the protamine or SNBP. Histone substitution causes chromatin remodelling and additional reduction in nuclear volume. We analyze these three cases together with earlier works and propose that the formation of 20 nm granules containing acetylated H3 and H4 accomplishes the minimum functional requirement to be considered the most evolutionarily ancestral chromatin conformation preceding condensation in animal spermiogenesis. PMID- 19132735 TI - Crown formation during tooth development and tissue engineering. AB - Considering tooth crown engineering, three main parameters have to be taken into account: (1) the relationship between crown morphology and tooth functionality, (2) the growth of the organ, which is hardly compatible with the use of preformed scaffolds, and (3) the need for easily available nondental competent cell sources. In vitro reassociation experiments using either dental tissues or bone marrow-derived cells (BMDC) have been designed to get information about the mechanisms to be preserved in order to allow crown engineering. As the primary enamel knot (PEK) is involved in signaling crown morphogenesis, the formation and fate of this structure was investigated (1) in heterotopic reassociations between embryonic day 14 (ED14) incisor and molar enamel organs and mesenchymes, and (2) in reassociations between ED14 molar enamel organs and BMDC. A PEK formed in cultured heterotopic dental tissue reassociations. The mesenchyme controls the fate of the EK cells, incisor or molar-specific using apoptosis as criterion, and functionality to drive single/multiple cusps tooth development. Although previous investigations showed that they might differentiate as odontoblast- or ameloblast like cells, BMDC reassociated to an enamel organ could not support the development of multicusp teeth. These cells apparently could neither maintain nor stimulate the formation of a PEK. PMID- 19132736 TI - Functional analysis of neurovascular adaptations to exercise in the dentate gyrus of young adult mice associated with cognitive gain. AB - The discovery that aerobic exercise increases adult hippocampal neurogenesis and can enhance cognitive performance holds promise as a model for regenerative medicine. This study adds two new pieces of information to the rapidly growing field. First, we tested whether exercise increases vascular density in the granular layer of the dentate gyrus, whole hippocampus, and striatum in C57BL/6J mice known to display procognitive effects of exercise. Second, we determined the extent to which new neurons from exercise participate in the acute neuronal response to high levels of running in B6D2F1/J (F1 hybrid of C57BL/6J female by DBA/2J male). Mice were housed with or without a running wheel for 50 days (runner vs. sedentary). The first 10 days, they received daily injections of BrdU to label dividing cells. The last 10 days, mice were tested for performance on the Morris water maze and rotarod and then euthanized to measure neurogenesis, c Fos induction from running and vascular density. In C57BL/6J, exercise increased neurogenesis, density of blood vessels in the dentate gyrus and striatum (but not whole hippocampus), and enhanced performance on the water maze and rotarod. In B6D2F1/J, exercise also increased hippocampal neurogenesis but not vascular density in the granular layer. Improvement on the water maze from exercise was marginal, and no gain was seen for rotarod, possibly because of a ceiling effect. Running increased the number of c-Fos positive neurons in the granular layer by fivefold, and level of running was strongly correlated with c-Fos within 90 min before euthanasia. In runners, approximately 3.3% (+/-0.008 S.E.) of BrdU positive neurons in the middle of the granule layer displayed c-Fos when compared with 0.8% (+/-0.001) of BrdU-negative neurons. Results suggest that procognitive effects of exercise are associated with increased vascular density in the dentate gyrus and striatum in C57BL/6J mice, and that new neurons from exercise preferentially function in the neuronal response to running in B6D2F1/J. PMID- 19132737 TI - Religion and spirituality in psychotherapy: a practice-friendly review of research. AB - The role of religion and spirituality in psychotherapy has received growing attention in the last two decades, with a focus on understanding the ways that religion and spirituality relate to therapists, clients, and treatment methods. The authors reviewed recent empirical research on religion and spirituality in psychotherapy to inform practitioners about effective ways to incorporate the sacred into their clinical work. Three main areas are covered: religion/spirituality and therapists, religion/spirituality and clients, and religious/spiritual interventions. Research indicates that therapists are open to religious/spiritual issues, that clients want to discuss these matters in therapy, and that the use of religious/spiritual interventions for some clients can be an effective adjunct to traditional therapy interventions. PMID- 19132738 TI - A theistic spiritual treatment for women with eating disorders. AB - The authors describe a psychological treatment for women with eating disorders who have theistic spiritual beliefs and illustrate its application with a case report. They begin by briefly summarizing a theistic view of eating disorders. Then they illustrate how a theistic approach can complement traditional treatment by describing the processes and outcomes of their work with a 23-year-old Christian woman receiving inpatient treatment for an eating disorder not otherwise specified and a major depressive disorder (recurrent severe). PMID- 19132739 TI - Integrating spirituality into alcohol treatment. AB - Spirituality is presumed by millions of Americans to be directly relevant to problems of alcohol abuse. We summarize findings regarding the role of religion and spirituality in the prevention and treatment of substance abuse and present a case illustration. We also consider mechanisms responsible for these effects. We offer advice about why, by whom, and how religion and spirituality should be discussed with clients with substance use disorders. In a recent clinical trial, therapists trained in a client-centered approach to facilitate exploration of spirituality fostered clients' use of spiritual practices. We suggest that the therapist's ability to skillfully engage clients in a discussion of spirituality is largely determined by how the therapist balances the dual roles of authoritative expert and evocative facilitator. PMID- 19132740 TI - Religiously oriented mindfulness-based cognitive therapy. AB - The interface of religiously accommodative and oriented treatments and the cognitive-behavioral tradition is explored. In terms of Hayes' characterization of the evolution of the cognitive-behavioral tradition through three waves, considerable theoretical, clinical, and empirical work emerged to support a religiously accommodative cognitive-behavioral therapy (CBT) during the second generation CBTs. Rather than including religion and spirituality, the third-wave CBT traditions have engaged in spiritual themes inspired heavily from Eastern religious traditions. The authors discuss the application of a religiously congruent third-wave cognitive therapy with a depressed conservatively Christian client. Some conceptual challenges and rationales for adopting such treatments with Christian or other theist clients are described. PMID- 19132742 TI - Biofilm monitoring on rotating discs by image analysis. AB - The macrostructure development of biofilms grown in a lab-scale rotating biological contactor was monitored by analyzing the average opacity and the texture of gray-level images of the discs. The reactor was fed with municipal or synthetic wastewater. Experiments lasted on average 4-14 weeks. The images were obtained with a flat-bed scanner. The opacity and its standard deviation are directly extracted from the annular zone where the biofilm develops. This zone is defined by the outer edge of the disc and the waterline. The spatial gray-level dependence matrix (SGLDM) approach was used for the texture assessment. As this method requires rectangular images, a geometrical transformation had to be developed to transform the ring into a workable area. This transformation now allows quantitative image analysis on circular biofilms. As a last step, Principal Components Analysis was applied to the set of textural descriptors to reduce the number of textural parameters. Opacity and textural information allowed the non-intrusive monitoring of the growth/regrowth of the biofilms as well as biofilm loss, due to detachment, auto-digestion, or protozoan grazing. Textural description was very valuable by helping to discriminate biofilms of similar opacity characteristics but presenting different macrostructures. PMID- 19132743 TI - Small molecule clearance in ultrafiltration/diafiltration in relation to protein interactions:Study of citrate binding to a Fab. AB - Ultrafiltration/diafiltration (UFDF) is commonly utilized in the purification of recombinant proteins to concentrate and buffer exchange the product. It is often the final step in the purification process, placing the protein in its final formulation and clearing small molecules introduced in upstream purification steps. This article presents a case study of reduced small molecule clearance in ultrafiltration/diafiltration of an antigen-binding fragment of a monoclonal antibody. Citrate, a commonly utilized small molecule in downstream processes, is shown to have reduced clearance due to specific interactions with the protein product. The study presents process solutions and utilizes a simple model to characterize clearance of small molecules which exhibit interactions with product protein. PMID- 19132744 TI - Mining transcriptome data for function-trait relationship of hyper productivity of recombinant antibody. AB - In the past decade we have witnessed a drastic increase in the productivity of mammalian cell culture-based processes. High-producing cell lines that synthesize and secrete these therapeutics have contributed largely to the advances in process development. To elucidate the productivity trait in the context of physiological functions, the transcriptomes of several NS0 cell lines with a wide range of antibody productivity were compared. Gene set testing (GST) analysis was used to identify pathways and biological functions that are altered in high producers. Three complementary tools for GST-gene set enrichment analysis (GSEA), gene set analysis (GSA), and MAPPFinder, were used to identify groups of functionally coherent genes that are up- or downregulated in high producers. Major functional classes identified include those involved in protein processing and transport, such as protein modification, vesicle trafficking, and protein turnover. A significant proportion of genes involved in mitochondrial ribosomal function, cell cycle regulation, cytoskeleton-related elements are also differentially altered in high producers. The observed correlation of these functional classes with productivity suggests that simultaneous modulation of several physiological functions is a potential route to high productivity. PMID- 19132745 TI - Monitoring of biomass composition from microbiological sources by means of FT-IR spectroscopy. AB - An FT-IR spectroscopic method was developed for the simultaneous quantitative analysis of biomacromolecular components in biomass, originating from various microbiological sources. For the determination of protein, lipid and carbohydrate content, creatine phosphokinase, egg phosphatidyl choline and starch hydrolysate were chosen as external standards. This selection was based on spectral similarity and ease of availability. Protein content was based on the area under the amide II band profile around 1,545 cm(-1). Because of the heterogeneous lipid composition in the different species, lipid content was determined using integration over the C-H stretching vibrational population between 2,984 and 2,780 cm(-1). Carbohydrate content was determined using integration over a C-O and C-O-C stretching band area between 1,180 and 1,133 cm(-1). Linear regression analysis provided three calibration lines, according to which biomasses from ten species were analyzed. This approach showed good intra-batch reproducibility. With this method we could demonstrate good reproducibility between batches of the same species with similar growth conditions while large differences in biomass composition were observed between the various species. Protein content as determined by FT-IR spectroscopy compared well with the results obtained from elemental analysis. PMID- 19132746 TI - On the role of protein structural dynamics in the catalytic activity and thermostability of serine protease subtilisin Carlsberg. AB - The effect of structural dynamics on enzyme activity and thermostability has thus far only been investigated in detail for the serine protease alpha-chymotrypsin (for a recent review see Sola et al., Cell Mol Life Sci 2007, 64(16): 2133-2152). Herein, we extend this type of study to a structurally unrelated serine protease, specifically, subtilisin Carlsberg. The protease was incrementally glycosylated with chemically activated lactose to obtain various subtilisin glycoconjugates which were biophysically characterized. Near UV-CD spectroscopy revealed that the tertiary structure was unaffected by the glycosylation procedure. H/D exchange FT IR spectroscopy was performed to assess the changes in structural dynamics of the enzyme. It was found that increasing the level of glycosylation caused a linearly dependent reduction in structural dynamics. This led to an increase in thermostability and a decrease in the catalytic turnover rate for both, the enzyme acylation and deacylation steps. These results highlight the possibility that a structural dynamics-activity relationship might be a phenomenon generally found in serine proteases. PMID- 19132747 TI - PMS2 involvement in patients suspected of Lynch syndrome. AB - It is well-established that germline mutations in the mismatch repair genes MLH1, MSH2, and MSH6 cause Lynch syndrome. However, mutations in these three genes do not account for all Lynch syndrome (suspected) families. Recently, it was shown that germline mutations in another mismatch repair gene, PMS2, play a far more important role in Lynch syndrome than initially thought. To explore this further, we determined the prevalence of pathogenic germline PMS2 mutations in a series of Lynch syndrome-suspected patients. Ninety-seven patients who had early-onset microsatellite instable colorectal or endometrial cancer, or multiple Lynch syndrome-associated tumors and/or were from an Amsterdam Criteria II-positive family were selected for this study. These patients carried no pathogenic germline mutation in MLH1, MSH2, or MSH6. When available, tumors were investigated for immunohistochemical staining (IHC) for PMS2. PMS2 was screened in all patients by exon-by-exon sequencing. We identified four patients with a pathogenic PMS2 mutation (4%) among the 97 patients we selected. IHC of PMS2 was informative in one of the mutation carriers, and in this case, the tumor showed loss of PMS2 expression. In conclusion, our study confirms the finding of previous studies that PMS2 is more frequently involved in Lynch syndrome than originally expected. PMID- 19132749 TI - Bayesian models identify specific lymphoproliferative disorders associated with hepatitis C virus infection. PMID- 19132750 TI - Murine osteosarcoma primary tumour growth and metastatic progression is maintained after marked suppression of serum insulin-like growth factor I. AB - The insulin-like growth factor I (IGF-I) signaling pathway has been shown to play an important role in several aspects of cancer biology, including metastasis. The aim of this study was to define the contribution of serum (endocrine) and local (tumour microenvironment) IGF-I on osteosarcoma tumour growth and metastasis, a cancer that is known to be dependent on the IGF-I axis. To test this hypothesis, we evaluated the primary tumour growth and metastatic progression of K7M2 murine osteosarcoma cells injected to a genetically engineered mouse [liver-specific IGF I deficient (LID)] in which serum IGF-I levels are reduced by 75%, while maintaining expression of IGF-I in normal tissues. We first demonstrated that IGF I in the tumour and the tumour-microenvironment were maintained in the LID mice. Within this designed model, there was no difference in primary tumour growth or in pulmonary metastasis in LID mice compared to control mice. Furthermore, there was no difference in the number or localization of single metastatic cells immediately after their arrival in the lungs of LID mice and control mice, as analysed by single cell video microscopy. Collectively, these data suggest that marked reduction in serum IGF-I is not sufficient to slow the progression of either primary or metastatic models of osteosarcoma. PMID- 19132751 TI - Recurrent chemical reactivations of EBV promotes genome instability and enhances tumor progression of nasopharyngeal carcinoma cells. AB - Nasopharyngeal carcinoma (NPC) is an endemic malignancy prevalent in South East Asia. Epidemiological studies have associated this disease closely with Epstein Barr virus (EBV) infection. Previous studies also showed that EBV reactivation is implicated in the progression of NPC. Thus, we proposed that recurrent reactivations of EBV may be important for its pathogenic role. In this study, NPC cell lines latently infected with EBV, NA and HA, and the corresponding EBV negative NPC cell lines, NPC-TW01 (TW01) and HONE-1, were treated with 12-O tetradecanoylphorbol-13-acetate (TPA) and sodium n-butyrate (SB) for lytic cycle induction. A single treatment with TPA/SB revealed that DNA double-strand breaks and formation of micronuclei (a marker for genome instability) were associated with EBV reactivation in NA and HA cells. Examination of EBV early genes had identified several lytic proteins, particularly EBV DNase, as potent activators that induced DNA double-strand breaks and contribute to genome instability. Recurrent reactivations of EBV in NA and HA cells resulted in a marked increase of genome instability. In addition, the degree of chromosomal aberrations, as shown by chromosome structural variants and DNA copy-number alterations, is proportional to the frequency of TPA/SB-induced EBV reactivation. Whereas these DNA abnormalities were limited in EBV-negative TW01 cells with mock or TPA/SB treatment, and were few in mock-treated NA cells. The invasiveness and tumorigenesis assays also revealed a profound increase in both characteristics of the repeatedly reactivated NA cells. These results suggest that recurrent EBV reactivations may result in accumulation of genome instability and promote the tumor progression of NPC. PMID- 19132752 TI - Minimizing misclassification of hormone users at mammography screening. AB - The aim of the study was to retrospectively determine the impact of comparing current mammograms with prior mammograms on risk of misclassification especially for hormone users. Data on mammography screening were retrieved for 1993-2005 from Fyn, Denmark. At first screen, two projections were made; at subsequent screens, one projection for fatty and two projections for mixed/dense breasts. Until June 3, 2002, 2-year-old mammograms were used for comparison, and later 4 year-old mammograms. Prescription drug data were used to identify hormone, hormone therapy (HT), use. False positive risk and interval cancer proportion dependency on age, hormone use, screen number, projection and prior mammogram were tested with logistic regression. Controlled for breast density, current HT users had a lower risk of a false positive test 0.69 (95%CI 0.55-0.86) and a lower interval cancer proportion 0.66 (95%CI 0.45-0.99) when 4-year-old instead of 2-year-old mammograms were used for comparison. The use of 4-year-old instead of 2-year-old mammograms for comparison lowered the risk of false positive test in never users, but otherwise age of comparison mammogram had no impact on classification of never and past users of HT. The study indicated that misclassification at screening mammography in current users of HT can be reduced considerably, when the screening mammograms are viewed with the mammograms taken 4 years earlier. It should be stressed that these results come from a single clinic, and replication in other observational and/or experimental studies is warranted. PMID- 19132753 TI - Compact spheroid formation by ovarian cancer cells is associated with contractile behavior and an invasive phenotype. AB - Ovarian cancer cells are present in malignant ascites both as individual cells and as multicellular spheroid aggregates. Although spheroid formation affords protection of cancer cells against some chemotherapeutic agents, it has not been established whether a relationship exists between invasive behavior and predisposition to spheroid formation. Aspects of spheroid formation, including cell-matrix adhesion, remodeling and contractility are characteristic myofibroblast-like behaviors associated with fibrosis that contribute to tumor growth and dissemination. We explored the possibility that cell behaviors that promote spheroid formation also facilitate invasion. Our analysis of 6 human ovarian cancer cell lines indicated that ovarian cancer cells possessing myofibroblast-like properties formed compact spheroids and invaded 3D matrices. These cells readily contracted collagen I gels, possessed a spindle-like morphology, and had elevated expression of genes associated with the TGFbeta mediated fibrotic response and/or beta1 integrin function, including fibronectin (FN), connective tissue growth factor (CTGF/CCN2), lysyl oxidase (LOX1), tissue transglutaminase 2 (TGM2) and urinary plasminogen activator receptor (uPAR). Whereas cell aggregation was induced by TGFbeta, and by beta1-integrin overexpression and activation, these treatments did not stimulate the contractile activity required for spheroid compaction. The positive relationship found between compact spheroid formation and invasive behavior implies a preferential survival of an invasive subpopulation of ovarian cancer cells, as cells in spheroids are more resistant to several chemotherapeutics. Preventing the formation of ovarian cancer spheroids may represent a novel strategy to improve the efficacy of existing therapeutics. PMID- 19132754 TI - Putative functional polymorphisms of MMP9 predict survival of NSCLC in a Chinese population. AB - Matrix metalloproteinases (MMPs) play a crucial role in cancer progression and their over-expression is often associated with unfavorable survival of non-small cell lung cancer (NSCLC). Because genetic variants can alter expression level or biological activity of MMPs, we hypothesized that potentially functional single nucleotide polymorphisms (SNPs) in key MMP genes may be associated with the survival of NSCLC patients. We selected and genotyped 14 putative functional SNPs in six MMP genes (MMP1, MMP2, MMP3, MMP7, MMP9 and MMP12) using PCR-RFLP methods in 561 NSCLC patients. Kaplan-Meier method with the log-rank test and Cox proportional hazard models were used for the survival analyses. The C-1562T, Arg279Gln and Arg668Gln polymorphisms in MMP9 were significantly associated with survival of patients with NSCLC (log-rank p values = 0.032, 0.038 and 0.036, respectively). The C-1562T and Arg668Gln loci were in complete linkage disequilibrium (r(2) = 1). Patients carrying the 668Gln allele had improved survival with a median survival time (MST) of 51.6 months, compared with 21.8 months for those with the 668Arg/Arg genotype (log-rank p = 0.010). In contrast, the 279Gln/Gln genotype was associated with a significantly shortened MST (17.3 months, log-rank p = 0.030) in the recessive model. In the final multivariate Cox regression model, 279Gln/Gln was identified as an independent prognostic factor with an adjusted hazard ratio of 1.60 (95% confidence interval 1.07-2.41). The MMP9 Arg279Gln and Arg668Gln SNPs are potential predictors of survival in NSCLC patients. PMID- 19132755 TI - Eddy-current compensated diffusion weighting with a single refocusing RF pulse. AB - A modification of the Stejskal-Tanner diffusion-weighting preparation with a single refocusing RF pulse is presented which involves three gradient lobes that can be adjusted to null eddy currents with any given decay rate to reduce geometric distortions in diffusion-weighted echo-planar imaging (EPI). It has a very similar compensation performance as the commonly used double-spin-echo preparation but (i) is less sensitive to flip angle imperfections, e.g. along the slice profile, and B(1) inhomogeneities and (ii) can yield shorter echo times for moderate b values, notably for longer echo trains as required for higher spatial resolution. It therefore can provide an increased signal-to-noise ratio as is simulated numerically and demonstrated experimentally in water phantoms and the human brain for standard EPI (2.0 x 2.0 mm(2)) and high-resolution EPI of inner field-of-views using 2D-selective RF excitations (0.5 x 1.0 mm(2)). Thus, the presented preparation may help to overcome current limitations of diffusion weighted EPI, in particular at high static magnetic fields. PMID- 19132756 TI - Functional changes in CSF volume estimated using measurement of water T2 relaxation. AB - Cerebrospinal fluid (CSF) provides hydraulic suspension for the brain. The general concept of bulk CSF production, circulation, and reabsorption is well established, but the mechanisms of momentary CSF volume variation corresponding to vasoreactive changes are far less understood. Nine individuals were studied in a 3T MR scanner with a protocol that included visual stimulation using a 10-Hz reversing checkerboard and administration of a 5% CO(2) mix in air. We acquired PRESS-localized spin-echoes (TR = 12 sec, TE = 26 ms to 1.5 sec) from an 8-mL voxel located in the visual cortex. Echo amplitudes were fitted to a two compartmental model of relaxation to estimate the partial volume of CSF and the T(2) relaxation times of the tissues. CSF signal contributed 10.7 +/- 3% of the total, with T(2,csf) = 503.0 +/- 64.3 [ms], T(2,brain) = 61.0 +/- 2 [ms]. The relaxation time of tissue increased during physiological stimulation, while the fraction of signal contributed by CSF decreased significantly by 5-6% with visual stimulation (P < 0.03) and by 3% under CO(2) inhalation (P < 0.08). The CSF signal fraction is shown to represent well the volume changes under viable physiological scenarios. In conclusion, CSF plays a significant role in buffering the changes in cerebral blood volume, especially during rapid functional stimuli. PMID- 19132757 TI - Variation in the shape of pulsed arterial spin labeling kinetic curves across the healthy human brain and its implications for CBF quantification. AB - Arterial spin labeling (ASL) uses proximally inverted blood water spins as an endogenous contrast agent to measure blood perfusion in tissue. In this work pulsed ASL was used to measure the kinetic curves of cerebral blood flow (CBF) across eight anatomically based regions of interest (ROIs) in five normal healthy subjects. CBF estimates based on these data were compared when obtained using a single inversion time versus fitting a kinetic model to all 10 measured inversion times. CBF estimates were also compared when fitting to the 10 inversion time data using a standard two-parameter approach (CBF and bolus arrival time) or a more recently proposed three-parameter model (CBF, bolus arrival time, and arrival time dispersion). Variations in the shape of the kinetic curve were found across the brain that were consistent across subjects. The arrival time in the occipital ROI was found to be long enough to lead to underestimation of the CBF when using a single inversion time of 1.5 s. In four out of the eight ROIs there was significant underestimation of CBF using the standard two-parameter model compared to the three-parameter approach. These results have important implications for the development of a robust, quantitative ASL protocol. PMID- 19132758 TI - Magnetic resonance elastography with a phased-array acoustic driver system. AB - Dynamic MR elastography (MRE) quantitatively maps the stiffness of tissues by imaging propagating shear waves in the tissue. These waves can be produced from intrinsic motion sources (e.g., due to cardiac motion), from external motion sources that produce motion directly at depth in tissue (e.g., amplitude modulated focused ultrasound), and from external actuators that produce motion at the tissue surface that propagates into the tissue. With external actuator setups, typically only a single transducer is used to create the shear waves, which in some applications might have limitations due to shadowing and attenuation of the waves. To address these limitations, a phased-array acoustic driver system capable of applying independently controlled waveforms to each channel was developed and tested. It was found that the system produced much more uniform illumination of the object, improving the quality of the elastogram. It was also found that the accuracy of the stiffness value of any arbitrary region of interest could be improved by obtaining maximal shear wave illumination with the phased array capability of the system. PMID- 19132759 TI - Measuring RF-induced currents inside implants: Impact of device configuration on MRI safety of cardiac pacemaker leads. AB - Radiofrequency (RF)-related heating of cardiac pacemaker leads is a serious concern in magnetic resonance imaging (MRI). Recent investigations suggest such heating to be strongly dependent on an implant's position within the surrounding medium, but this issue is currently poorly understood. In this study, phantom measurements of the RF-induced electric currents inside a pacemaker lead were performed to investigate the impact of the device position and lead configuration on the amount of MRI-related heating at the lead tip. Seven hundred twenty device position/lead path configurations were investigated. The results show that certain configurations are associated with a highly increased risk to develop MRI induced heating, whereas various configurations do not show any significant heating. It was possible to precisely infer implant heating on the basis of current intensity values measured inside a pacemaker lead. Device position and lead configuration relative to the surrounding medium are crucial to the amount of RF-induced heating in MRI. This indicates that a considerable number of implanted devices may incidentally not develop severe heating in MRI because of their specific configuration in the body. Small variations in configuration can, however, strongly increase the risk for such heating effects, meaning that hazardous situations might appear during MRI. PMID- 19132760 TI - New technologies in gastrointestinal research. AB - This issue presents different new techniques aiming to increase our understanding of the gastrointestinal system and to improve treatment. The technologies cover selected methods to evoke and assess gut pain, new methods for imaging and physiological measurements, histochemistry, pharmacological modelling etc. There is no doubt that the methods will revolutionize the diagnostic approach in near future. PMID- 19132761 TI - New technologies in the gastrointestinal clinic and research: impedance and high resolution manometry. AB - The last five years have been an exciting time in the study of esophageal motor disorders due to the recent advances in esophageal function testing. New technologies have emerged, such as intraluminal impedance, while conventional techniques, such as manometry, have enjoyed many improvements due to advances in transducer technology, computerization and graphic data presentation. While these techniques provide more detailed information regarding esophageal function, our understanding of whether they can improve our ability to diagnose and treat patients more effectively is evolving. These techniques are also excellent research tools and they have added substantially to our understanding of esophageal motor function in dysphagia. This review describes the potential benefits that these new technologies may have over conventional techniques for the evaluation of dysphagia. PMID- 19132762 TI - Axial force measurement for esophageal function testing. AB - The esophagus serves to transport food and fluid from the pharynx to the stomach. Manometry has been the "golden standard" for the diagnosis of esophageal motility diseases for many decades. Hence, esophageal function is normally evaluated by means of manometry even though it reflects the squeeze force (force in radial direction) whereas the bolus moves along the length of esophagus in a distal direction. Force measurements in the longitudinal (axial) direction provide a more direct measure of esophageal transport function. The technique used to record axial force has developed from external force transducers over in-vivo strain gauges of various sizes to electrical impedance based measurements. The amplitude and duration of the axial force has been shown to be as reliable as manometry. Normal, as well as abnormal, manometric recordings occur with normal bolus transit, which have been documented using imaging modalities such as radiography and scintigraphy. This inconsistency using manometry has also been documented by axial force recordings. This underlines the lack of information when diagnostics are based on manometry alone. Increasing the volume of a bag mounted on a probe with combined axial force and manometry recordings showed that axial force amplitude increased by 130% in contrast to an increase of 30% using manometry. Using axial force in combination with manometry provides a more complete picture of esophageal motility, and the current paper outlines the advantages of using this method. PMID- 19132763 TI - Do we really understand the role of the oesophagogastric junction in disease? AB - The role of the oesophago-gastric junction (OGJ) in gastro-oesophageal reflux disease is still not completely understood, and there is no clinically used method to assess the OGJ function in patients. Only indirect methods such as pH studies are carried out. The OGJ acts a valve controlling the flow of solids, liquids and gases between the oesophagus and the stomach. Manometry can determine if a sphincter is toned or relaxed; but, it cannot confirm that the sphincter region is actually open. Distension is a new technique for measuring function in the OGJ. By measuring the cross-sectional area through the narrow region in the junction during distension of a catheter mounted bag, much more information on the opening and closing patterns of the junction can be determined. This technique has already been demonstrated to show changes in the OGJ after surgical treatments for reflux disease. New measurement ideas around the concept of distending the OGJ offer new hope that a clinically useable test for compliance at the junction can be developed and could potentially help in determining appropriate therapy. PMID- 19132764 TI - Sensory testing of the human gastrointestinal tract. AB - The objective of this appraisal is to shed light on the various approaches to screen sensory information in the human gut. Understanding and characterization of sensory symptoms in gastrointestinal disorders is poor. Experimental methods allowing the investigator to control stimulus intensity and modality, as well as using validated methods for assessing sensory response have contributed to the understanding of pain mechanisms. Mechanical stimulation based on impedance planimetry allows direct recordings of luminal cross-sectional areas, and combined with ultrasound and magnetic resonance imaging, the contribution of different gut layers can be estimated. Electrical stimulation depolarizes free nerve endings non-selectively. Consequently, the stimulation paradigm (single, train, tetanic) influences the involved sensory nerves. Visual controlled electrical stimulation combines the probes with an endoscopic approach, which allows the investigator to inspect and obtain small biopsies from the stimulation site. Thermal stimulation (cold or warm) activates selectively mucosal receptors, and chemical substances such as acid and capsaicin (either alone or in combination) are used to evoke pain and sensitization. The possibility of multimodal (e.g. mechanical, electrical, thermal and chemical) stimulation in different gut segments has developed visceral pain research. The major advantage is involvement of distinctive receptors, various sensory nerves and different pain pathways mimicking clinical pain that favors investigation of central pain mechanisms involved in allodynia, hyperalgesia and referred pain. As impairment of descending control mechanisms partly underlies the pathogenesis in chronic pain, a cold pressor test that indirectly stimulates such control mechanisms can be added. Hence, the methods undoubtedly represent a major step forward in the future characterization and treatment of patients with various diseases of the gut, which provides knowledge to clinicians about the underlying symptoms and treatment of these patients. PMID- 19132765 TI - Imaging of the gastrointestinal tract-novel technologies. AB - Imaging of the gastrointestinal tract is very useful for research and clinical studies of patients with symptoms arising from the gastrointestinal tract and in visualising anatomy and pathology. Traditional radiological techniques played a leading role in such studies for a long time. However, advances in non-invasive modalities including ultrasound (US), computed tomography (CT), positron emission tomography (PET), magnetic resonance imaging (MRI), etc, have in the last decades revolutionised the way in which the gastrointestinal tract is studied. The resolution of imaging data is constantly being improved and 3D acquisition, tools for filtering, enhancement, segmentation and tissue classification are continually being developed. Additional co-registration techniques allow multimodal data acquisition with improved classification of tissue pathology. Furthermore, new functional imaging techniques have become available. Altogether, the future of gastrointestinal imaging looks very promising which will be of great benefit in clinical and research studies of gastrointestinal diseases. The purpose of this review is to highlight the capabilities of the newest techniques to explore the detailed morphology, biomechanical properties, function and pathology of the gastrointestinal tract. PMID- 19132766 TI - Gastrointestinal tract modelling in health and disease. AB - The gastrointestinal (GI) tract is the system of organs within multi-cellular animals that takes in food, digests it to extract energy and nutrients, and expels the remaining waste. The various patterns of GI tract function are generated by the integrated behaviour of multiple tissues and cell types. A thorough study of the GI tract requires understanding of the interactions between cells, tissues and gastrointestinal organs in health and disease. This depends on knowledge, not only of numerous cellular ionic current mechanisms and signal transduction pathways, but also of large scale GI tissue structures and the special distribution of the nervous network. A unique way of coping with this explosion in complexity is mathematical and computational modelling; providing a computational framework for the multilevel modelling and simulation of the human gastrointestinal anatomy and physiology. The aim of this review is to describe the current status of biomechanical modelling work of the GI tract in humans and animals, which can be further used to integrate the physiological, anatomical and medical knowledge of the GI system. Such modelling will aid research and ensure that medical professionals benefit, through the provision of relevant and precise information about the patient's condition and GI remodelling in animal disease models. It will also improve the accuracy and efficiency of medical procedures, which could result in reduced cost for diagnosis and treatment. PMID- 19132767 TI - Translational pain research: evaluating analgesic effect in experimental visceral pain models. AB - Deep visceral pain is frequent and presents major challenges in pain management, since its pathophysiology is still poorly understood. One way to optimize treatment of visceral pain is to improve knowledge of the mechanisms behind the pain and the mode of action of analgesic substances. This can be achieved through standardized experimental human pain models. Experimental pain models in healthy volunteers are advantageous for evaluation of analgesic action, as this is often difficult to assess in the clinic because of confounding factors such as sedation, nausea and general malaise. These pain models facilitate minimizing the gap between knowledge gained in animal and human clinical studies. Combining experimental pain studies and pharmacokinetic studies can improve understanding of the pharmacokinetic-pharmacodynamic relationship of analgesics and, thus, provide valuable insight into optimal clinical treatment of visceral pain. To improve treatment of visceral pain, it is important to study the underlying mechanisms of pain and the action of analgesics used for its treatment. An experimental pain model activates different modalities and can be used to investigate the mechanism of action of different analgesics in detail. In combination with pharmacokinetic studies and objective assessment such as electroencephalography, new information regarding a given drug substance and its effects can be obtained. Results from experimental human visceral pain research can bridge the gap in knowledge between animal studies and clinical condition in patients suffering from visceral pain, and thus constitute the missing link in translational pain research. PMID- 19132768 TI - New technologies to investigate the brain-gut axis. AB - Functional gastrointestinal disorders are commonly encountered in clinical practice, and pain is their commonest presenting symptom. In addition, patients with these disorders often demonstrate a heightened sensitivity to experimental visceral stimulation, termed visceral pain hypersensitivity that is likely to be important in their pathophysiology. Knowledge of how the brain processes sensory information from visceral structures is still in its infancy. However, our understanding has been propelled by technological imaging advances such as functional Magnetic Resonance Imaging, Positron Emission Tomography, Magnetoencephalography, and Electroencephalography (EEG). Numerous human studies have non-invasively demonstrated the complexity involved in functional pain processing, and highlighted a number of subcortical and cortical regions involved. This review will focus on the neurophysiological pathways (primary afferents, spinal and supraspinal transmission), brain-imaging techniques and the influence of endogenous and psychological processes in healthy controls and patients suffering from functional gastrointestinal disorders. Special attention will be paid to the newer EEG source analysis techniques. Understanding the phenotypic differences that determine an individual's response to injurious stimuli could be the key to understanding why some patients develop pain and hyperalgesia in response to inflammation/injury while others do not. For future studies, an integrated approach is required incorporating an individual's psychological, autonomic, neuroendocrine, neurophysiological, and genetic profile to define phenotypic traits that may be at greater risk of developing sensitised states in response to gut inflammation or injury. PMID- 19132769 TI - New techniques in the tissue diagnosis of gastrointestinal neuromuscular diseases. AB - Gastrointestinal neuromuscular diseases are a clinically heterogeneous group of disorders of children and adults in which symptoms are presumed or proven to arise as a result of neuromuscular (including interstitial cell of Cajal) dysfunction. Common to most of these diseases are symptoms of impaired motor activity which manifest as slowed or obstructed transit with or without evidence of transient or persistent radiological visceral dilatation. A variety of histopathological techniques and allied investigations are being increasingly applied to tissue biopsies from such patients. This review outlines some of the more recent advances in this field, particularly in the most contentious area of small bowel disease manifesting as intestinal pseudo-obstruction. PMID- 19132770 TI - Mucosal blood flow measurements using laser Doppler perfusion monitoring. AB - Perfusion of individual tissues is a basic physiological process that is necessary to sustain oxygenation and nutrition at a cellular level. Ischemia, or the insufficiency of perfusion, is a common mechanism for tissue death or degeneration, and at a lower threshold, a mechanism for the generation of sensory signalling including pain. It is of considerable interest to study perfusion of peripheral abdominal tissues in a variety of circumstances. Microvascular disease of the abdominal organs has been implicated in the pathogenesis of a variety of disorders, including peptic ulcer disease, inflammatory bowel disease and chest pain. The basic principle of laser Doppler perfusion monitoring (LDPM) is to analyze changes in the spectrum of light reflected from tissues as a response to a beam of monochromatic laser light emitted. It reflects the total local microcirculatory blood perfusion, including perfusion in capillaries, arterioles, venules and shunts. During the last 20-25 years, numerous studies have been performed in different parts of the gastrointestinal (GI) tract using LDPM. In recent years we have developed a multi-modal catheter device which includes a laser Doppler probe, with the intent primarily to investigate patients suffering from functional chest pain of presumed oesophageal origin. Preliminary studies show the feasibility of incorporating LDPM into such catheters for performing physiological studies in the GI tract. LDPM has emerged as a research and clinical tool in preference to other methods; but, it is important to be aware of its limitations and account for them when reporting results. PMID- 19132771 TI - Prevalence of vacA, cagA and babA2 genes in Cuban Helicobacter pylori isolates. AB - AIM: To investigate the prevalence of vacuolating cytotoxin (vacA), cytotoxin associated gene A (cagA) and blood adhesion binding antigen (babA2) genotypes of Helicobacter pylori (H pylori) isolates from Cuban dyspeptic patients. METHODS: DNA was extracted from H pylori-positive cultures taken from 130 dyspeptic patients. Genotyping was performed by PCR, using specific primers for vacA (s1, s2, m1, m2), cagA and babA2 genes. Endoscopic observations and histological examinations were used to determine patient pathologies. RESULTS: vacA alleles s1, s2, m1 and m2 were detected in 96 (73.8%), 34 (26.2%), 75 (57.7%) and 52 isolates (40%), respectively, while the cagA gene was detected in 95 isolates (73.2%). One hundred and seven isolates (82.3%) were babA2-positive. A significant correlation was observed between vacAs1m1 and cagA and between vacAs1m1 and babA2 genotypes (P < 0.001 and P < 0.05, respectively) and between babA2 genotype and cagA status (P < 0.05); but, no correlation was observed between vacAs1 and babA2 genotypes. Eighty five (65.4%) and 73 (56.2%) strains were type 1 (vacAs1-cagA-positive) and "triple-positive" (vacAs1-cagA-babA2 positive), respectively, and their presence was significantly associated with duodenal ulcer (P < 0.01 and P < 0.001, respectively). CONCLUSION: The distribution of the main virulence factors in the Cuban strains in this study resembled that of the Western-type strains, and the more virulent H pylori isolates were significantly associated with duodenal ulcer, ulcer disease being the worst pathology observed in the group studied. PMID- 19132772 TI - Role of bacterial and genetic factors in gastric cancer in Costa Rica. AB - AIM: To evaluate several risk factors for gastric cancer (GC) in Costa Rican regions with contrasting GC incidence rate (GCIR). METHODS: According to GCIR, 191 Helicobacter pylori (H pylori)-positive patients were classified into groups A (high GCIR, n = 101) and B (low GCIR, n = 90). Human DNA obtained from biopsy specimens was used in the determination of polymorphisms of the genes coding for interleukin (IL)-1beta and IL-10 by PCR-RFLP, and IL-1RN by PCR. H pylori DNA extractions obtained from clinical isolates of 83 patients were used for PCR based genotyping of H pylori cagA, vacA and babA2. Human DNA from gastric biopsies of 52 GC patients was utilized for comparative purposes. RESULTS: Cytokine polymorphisms showed no association with GCIR variability. However, gastric atrophy, intestinal metaplasia and strains with different vacA genotypes in the same stomach (mixed strain infection) were more frequently found in group A than in group B, and cagA and vacA s1b were significantly associated with high GCIR (P = 0.026 and 0.041, respectively). IL-1beta+3954_T/C (OR 2.1, 1.0-4.3), IL 1RN*2/L (OR 3.5, 1.7-7.3) and IL-10-592_C/A (OR 3.2, 1.5-6.8) were individually associated with GC, and a combination of these cytokine polymorphisms with H pylori vacA s1b and m1 further increased the risk (OR 7.2, 1.4-36.4). CONCLUSION: Although a proinflammatory cytokine genetic profile showed an increased risk for developing GC, the characteristics of H pylori infection, in particular the status of cagA and vacA genotype distribution seemed to play a major role in GCIR variability in Costa Rica. PMID- 19132773 TI - Acute effects of Helicobacter pylori extracts on gastric mucosal blood flow in the mouse. AB - AIM: To investigate the mechanisms underlying the reduction in gastric blood flow induced by a luminal water extract of Helicobacter pylori (HPE). METHODS: The stomachs of isoflurane-anesthetized mice were exteriorized, and the mucosal surface exposed. Blood flow was measured with the laser-Doppler technique, and systemic arterial blood pressure monitored. C57BL/6 mice were exposed to water extract produced from H pylori strain 88-23. To investigate the role of a nerve- or iNOS-mediated pathway, we used intraluminal lidocaine and iNOS-/- mice. Blood flow response to the endogenous nitric oxide synthase inhibitor asymmetric dimethyl arginine (ADMA) was also assessed. RESULTS: In wild-type mice, HPE decreased mucosal blood flow by approximately 30%. This reduction was abolished in iNOS-deficient mice, and by pre-treatment with lidocaine. Luminally applied ADMA resulted in reduction in blood flow similar to that observed in wild-type mice exposed to HPE. CONCLUSION: A H pylori water extract reduces gastric mucosal blood flow acutely through iNOS- and nerve-mediated pathways. PMID- 19132774 TI - Adherence to surveillance guidelines for dysplasia and colorectal carcinoma in ulcerative and Crohn's colitis patients in the Netherlands. AB - AIM: To study adherence to the widely accepted surveillance guidelines for patients with long-standing colitis in The Netherlands. METHODS: A questionnaire was sent to all 244 gastroenterologists in The Netherlands. RESULTS: The response rate was 63%. Of all gastroenterologists, 95% performed endoscopic surveillance in ulcerative colitis (UC) patients and 65% in patients with Crohn's colitis. The American Gastroenterological Association (AGA) guidelines were followed by 27%, while 27% and 46% followed their local hospital protocol or no specific protocol, respectively. The surveillance was correctly initiated in cases of pancolitis by 53%, and in cases of left-sided colitis by 44% of the gastroenterologists. Although guidelines recommend 4 biopsies every 10 cm, less than 30 biopsies per colonoscopy were taken by 73% of the responders. Only 31%, 68% and 58% of the gastroenterologists referred patients for colectomy when low-grade dysplasia, high-grade dysplasia (HGD) or Dysplasia Associated Lesion or Mass (DALM) was present, respectively. CONCLUSION: Most Dutch gastroenterologists perform endoscopic surveillance without following international recommended guidelines. This practice potentially leads to a decreased sensitivity for dysplasia, rendering screening for colorectal cancer in this population highly ineffective. PMID- 19132775 TI - Features of hepatocellular carcinoma in cases with autoimmune hepatitis and primary biliary cirrhosis. AB - AIM: To characterize the clinical features of hepatocellular carcinoma (HCC) associated with autoimmune liver disease, we critically evaluated the literature on HCC associated with autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC). METHODS: A systematic review of the literature was conducted using the Japana Centra Revuo Medicina database which produced 38 cases of HCC with AIH (AIH-series) and 50 cases of HCC with PBC (PBC-series). We compared the clinical features of these two sets of patients with the general Japanese HCC population. RESULTS: On average, HCC was more common in men than in women with AIH or PBC. While many patients underwent chemolipiodolization (CL) or transcatheter arterial embolization (TAE) (AIH-series: P = 0.048 (vs operation), P = 0.018 (vs RFA, PEIT); PBC-series: P = 0.027 (vs RFA, PEIT), others refused therapeutic interventions [AIH-series: P = 0.038 (vs RFA, PEIT); PBC-series: P = 0.003 (vs RFA, PEIT)]. Liver failure was the primary cause of death among patients in this study, followed by tumor rupture. The survival interval between diagnosis and death was fairly short, averaging 14 +/- 12 mo in AIH patients and 8.4 +/- 14 mo in PBC patients. CONCLUSION: We demonstrated common clinical features among Japanese cases of HCC arising from AIH and PBC. PMID- 19132776 TI - Interaction of hepatitis C virus envelope glycoprotein E2 with the large extracellular loop of tupaia CD81. AB - AIM: To further analyze the interaction of tupaia CD81 with hepatitis C virus (HCV) envelope protein E2. METHODS: A tupaia CD81 large extracellular loop (CD81 LEL), which binds to HCV E2 protein, was cloned and expressed as a GST-fusion protein, and interaction of HCV E2 protein with a tupaia CD81 LEL was evaluated by enzyme-linked immunosorbent assay (EIA). RESULTS: Although tupaia and human CD81 LEL differed in 6 amino acid changes, tupaia CD81 LEL was strongly recognized by anti-CD81 antibodies against human CD81 LEL conformation-dependent epitopes. Investigating LEL CD81-E2 interactions by EIA, we demonstrated that binding of tupaia CD81 LEL GST fusion protein to recombinant HCV E2 protein was markedly reduced compared to binding of human CD81 LEL GST fusion protein to recombinant HCV E2 protein. CONCLUSION: These data suggest that the structural differences in-between the tupaia and human CD81 may alter the interaction of the large extracellular loop with HCV envelope glycoprotein E2. These findings may be important for the understanding of the mechanisms of binding and entry of HCV to PTHs. PMID- 19132777 TI - Perigastric extraskeletal Ewing's sarcoma: a case report. AB - Ewing's sarcoma (ES) is a neoplasm of undifferentiated small round cells, which occurs in the bones and deep soft tissues of children and adolescents. We present a rare case of a 44-year-old woman with gastric ES presenting with epigastric pain and weight loss. Ultrasound and computed tomography scans indicated a solid/cystic mass in the pancreatic tail. At laparotomy, the tumor was found attached to the posterior surface of the stomach, completely free from the pancreas, with no lymphadenopathy or local metastases. The polynodal, partly pseudocystic, dark-red soft tumor was excised. Histopathology revealed an anaplastic small-round-cell tumor with strong membranous CD99 immunoexpression. Additionally, there was patchy immunostaining for S-100 protein, vimentin, protein gene product (PGP) 9.5 and neuron-specific enolase, and weak focal CD117 cytoplasmic immunoreactivity. The patient had no adjuvant chemotherapy; her postoperative recovery was uneventful, and she remains symptom-free, and without any sign of recurrence at 20 mo. To the best of our knowledge, this is only the third ever case of gastric ES. PMID- 19132778 TI - A case report of endocrine cell carcinoma in the sigmoid colon with inferior mesenteric vein tumor embolism. AB - We report a case of endocrine cell carcinoma in the sigmoid colon with inferior mesenteric vein (IMV) tumor embolism. A 79-year-old woman was admitted to our hospital with narrowing of the stools. We performed colonoscopy, computed tomography and positron emission tomography, which disclosed sigmoid colon cancer with IMV tumor embolism. She underwent sigmoidectomy and lymph node dissection. The tumor was diagnosed as endocrine cell carcinoma (type 4, pSS, med, INFalpha, v3, n1, stage IIIb). Immunohistochemically, chromographin A, synaptophysin, cytokeratin 20 and mucicarmine showed partial staining, and CD56 was totally reactive. Three months after operation multiple liver metastases appeared. She was treated with chemotherapy of cisplatin (CDDP) + irinotecan (CPT11). This case highlights the aggressiveness of endocrine cell carcinoma with tumor embolism, and it is essential to establish an accurate diagnosis and effective treatment. PMID- 19132779 TI - Access to pediatric rheumatology subspecialty care in British Columbia, Canada. AB - OBJECTIVE: Early recognition and treatment of pediatric rheumatic diseases is associated with improved outcome. We documented access to pediatric rheumatology subspecialty care for children in British Columbia (BC), Canada, referred to the pediatric rheumatology clinic at BC Children's Hospital, Vancouver. METHODS: An audit of new patients attending the outpatient clinic from May 2006 to February 2007 was conducted. Parents completed a questionnaire through a guided interview at the initial clinic assessment. Referral dates were obtained from the referral letters. Patients were classified as having rheumatic disease, nonrheumatic disease, or a pain syndrome based on final diagnosis by a pediatric rheumatologist. RESULTS: Data were collected from 124 of 203 eligible new patients. Before pediatric rheumatology assessment, a median of 3 healthcare providers were seen (range 1-11) for a median of 5 visits (range 1-39). Overall, the median time interval from symptom onset to pediatric rheumatology assessment was 268 days (range 13-4989), and the median time interval from symptom onset to referral to pediatric rheumatology was 179 days (range 3-4970). Among patients ultimately diagnosed with rheumatic diseases (n = 53), there was a median of 119 days (range 3-4970) from symptom onset to referral, and 169 days (range 31-4989) from onset to pediatric rheumatology assessment. CONCLUSION: Children and adolescents with rheumatic complaints see multiple care providers for multiple visits before referral to pediatric rheumatology, and there is often a long interval between symptom onset and this referral. PMID- 19132780 TI - Effectiveness of specific neck stabilization exercises or a general neck exercise program for chronic neck disorders: a randomized controlled trial. AB - OBJECTIVE: In a cohort of primary care patients with chronic neck pain, to determine whether specific neck stabilization exercises, in addition to general neck advice and exercise, provide better clinical outcome at 6 weeks than general neck advice and exercise alone. METHODS: This was a multicenter randomized controlled trial in 4 physical therapy departments. Seventy-four participants (mean age 51.3 yrs) were randomized to specific neck stabilization exercises with a general neck advice and exercise program (n = 37) or a general neck advice and exercise program alone (n = 37). They attended a 1-hour clinical examination, followed by a maximum of 4 treatment sessions. Assessments were undertaken at baseline, 6 weeks, and 6 months. The primary outcome was the Neck Pain and Disability Scale (NPDS). Analysis was by intention to treat. RESULTS: Seventy-one (96%) participants received their allocated intervention. There was 91% followup at 6 weeks and 92% followup at 6 months. The mean (SD) 6-week improvement (reduction) in NPDS score was 10.6 (20.2) for the specific exercise program and 9.3 (15.7) for the general exercise program. There were no significant between group differences in the NPDS at either 6 weeks or 6 months. For secondary outcomes, participants in the specific exercise group were less likely to be taking pain medication at 6-week followup (p = 0.02). There were no other significant between-group differences. CONCLUSION: Adding specific neck stabilization exercises to a general neck advice and exercise program did not provide better clinical outcome overall in the physical therapy treatment of chronic neck pain. PMID- 19132781 TI - The efficacy and safety of milnacipran for treatment of fibromyalgia. a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the safety and efficacy of milnacipran, a dual norepinephrine and serotonin reuptake inhibitor, in the treatment of fibromyalgia (FM). METHODS: A 27-week, randomized, double-blind, multicenter study compared milnacipran 100 and 200 mg/day with placebo in the treatment of 888 patients with FM. Two composite responder definitions were used to classify each patient's individual response to therapy. "FM responders" concurrently satisfied response criteria for improvements in pain (visual analog scale 24-h morning recall), patient global impression of change (PGIC), and physical functioning (SF-36 Physical Component Summary); while "FM pain responders" concurrently satisfied response criteria for improvements in pain and PGIC. RESULTS: At the primary endpoint, after 3-month stable dose treatment, a significantly higher percentage of milnacipran-treated patients met criteria as FM responders versus placebo (milnacipran 200 mg/day, p = 0.017; milnacipran 100 mg/day, p = 0.028). A significantly higher percentage of patients treated with milnacipran 200 mg/day also met criteria as FM pain responders versus placebo (p = 0.032). Significant pain reductions were observed after Week 1 with both milnacipran doses. At 15 weeks, milnacipran 200 mg/day led to significant improvements over placebo in pain (realtime, daily and weekly recall; all measures, p < 0.05), PGIC (p < 0.001), fatigue (p = 0.016), cognition (p = 0.025), and multiple SF-36 domains. Milnacipran was safe and well tolerated by the majority of patients during 27 weeks of treatment; nausea and headache were the most common adverse events. CONCLUSION: Milnacipran is safe and effective for the treatment of multiple symptoms of FM. PMID- 19132782 TI - Dexamethasone promotes calcium pyrophosphate dihydrate crystal formation by articular chondrocytes. AB - OBJECTIVE: Calcium pyrophosphate dihydrate (CPPD) crystals are commonly found in osteoarthritic joints and correlate with a poor prognosis. Intraarticular corticosteroids, such as dexamethasone (Dxm), are commonly used therapies for osteoarthritis with or without CPPD deposition. Dxm has variable effects in mineralization models. We investigated the effects of Dxm on CPPD crystal formation in a well established tissue culture model. METHODS: Porcine articular chondrocytes were incubated with ATP to generate CPPD crystals. Chondrocytes incubated with or without ATP were exposed to 1-100 nM Dxm in the presence of 45Ca. Mineralization was measured by 45Ca uptake in the cell layer. We also investigated the effect of Dxm on mineralization-regulating enzymes such as alkaline phosphatase, nucleoside triphosphate pyrophosphohydrolase (NTPPPH), and transglutaminase. RESULTS: Dxm significantly increased ATP-induced mineralization by articular chondrocytes. While alkaline phosphatase and NTPPPH activities were unchanged by Dxm, transglutaminase activity increased in a dose-responsive manner. Levels of Factor XIIIA mRNA and protein were increased by Dxm, while type II Tgase protein was unchanged. Transglutaminase inhibitors suppressed Dxminduced increases in CPPD crystal formation. CONCLUSION: These findings suggest a potential for Dxm to contribute to pathologic mineralization in cartilage and reinforce a central role for the transglutaminase enzymes in CPPD crystal formation. PMID- 19132783 TI - Cementoplasty in the treatment of avascular necrosis of the hip. AB - OBJECTIVE: This retrospective study evaluated the role of percutaneous cementoplasty in the treatment of avascular necrosis (AVN) of the hip in order to postpone or avoid total hip replacement. METHODS: The study population comprised 40 patients (47 hips) with mean age of 46 +/- 4.7 years and mean body mass index of 26.7 +/- 4.6 kg/m(2). AVN was classified according to the Ficat-Arlet classification as one stage I, 30 stage II, and 16 stage III. The minimum followup was 9 months. RESULTS: It was found that 74.5% of hips were secondarily operated for total hip replacement a mean of 19.9 +/- 15 months (median 14 mo) after cementoplasty. As well, 94% of patients with stage 3AVN and 68% with stage 2AVN underwent surgery. Twelve hips were not operated, with a mean followup of 39 +/- 19.2 months. Pain decreased by more than 80% after cementoplasty in two thirds of patients, but the mean pain-free interval was only 8.1 +/- 6.6 months (median 5 mo). Nineteen of the 29 working patients were able to transiently return to work. The outcome was more unfavorable with radiological stage III AVN, joint effusion, and/or a double-line sign around the lesions on magnetic resonance images. CONCLUSION: Despite early relief of pain, the results of the cementoplasty technique were disappointing, with need for arthroplasty surgery in most cases within 2 years. Alternative percutaneous techniques using different filler materials with osteoinductive properties should be evaluated in further studies. PMID- 19132784 TI - Uveitis subtypes in a german interdisciplinary uveitis center--analysis of 1916 patients. AB - OBJECTIVE: Studies on the epidemiology of uveitis are rare and cohorts are small. We analyzed the frequencies of classified forms of uveitis in all patients at our center. METHODS: We studied 1916 consecutive patients with inflammatory eye disease. Data were analyzed regarding associated systemic disease, infection, ocular syndromes, anatomic localization, age, and sex. RESULTS: In 59.1% of patients, a classified form of uveitis was observed: associated systemic diseases in 43.7%, the most frequent ones sarcoidosis (17.4%) and ankylosing spondylitis (16.8%); ocular syndromes in 34.3%, the most frequent HLA-B27-positive anterior uveitis (AU; 35.1%) and Fuchs uveitis syndrome (FUS; 34.3%); and infections in 22.4%, the most frequent herpetic infections (46.1%) and toxoplasmosis (31.5%). We found AU in 45.4% of patients (15.4% HLA-B27-positive AU and 11.3% FUS), intermediate uveitis in 22.9% (unclassified 53.7% and multiple sclerosis 10.3%), and posterior uveitis in 13.5% (24.7% toxoplasmosis). Panuveitis was diagnosed in 6.2% of cases (Behcet's disease 12.6%; sarcoidosis 10.9%). The remaining 12.0% of cases showed extrauveal manifestations (scleritis, episcleritis, keratitis, optic neuritis, myositis, and orbital inflammation). CONCLUSION: We describe the largest cohort to date of consecutive patients from a specialized uveitis center. The high frequency of classified disease, nearly 60% in our clinic, shows the usefulness of an interdisciplinary approach, oriented on anatomic presentation. PMID- 19132785 TI - Determination of the subset of Sjogren's syndrome with articular manifestations by anticyclic citrullinated peptide antibodies. AB - OBJECTIVE: To investigate whether anticyclic citrullinated peptide antibodies (anti-CCP) predict the subset of Japanese patients with Sjogren's syndrome (SS) with articular manifestations. METHODS: Eighty-seven patients with SS were enrolled. Prevalence of anti-CCP antibodies, IgM rheumatoid factor, anti-Ro/SSA antibody, anti-La/SSB antibody, and serum IgG concentration and their relation to articular manifestations were examined. Articular manifestations included morning stiffness and the presence of tender or swollen joints. RESULTS: Eighty-seven SS patients were divided into 3 groups: 14 secondary SS with nonerosive rheumatoid arthritis (RA); 47 primary SS with articular manifestations; and 26 primary SS without articular manifestations. Ten out of 14 secondary SS with nonerosive RA expressed anti-CCP. Anti-CCP was the only statistically proven marker preferentially distributed in patients with articular manifestations (the first 2 groups) compared to primary SS without such manifestations; however, its frequency was low in primary SS. No patient with primary SS without articular manifestations expressed anti-CCP. CONCLUSION: Anti-CCP is found in the subset of Japanese with SS with articular manifestations although most of those with anti CCP-positive SS were classified as secondary SS with RA. PMID- 19132786 TI - Angiotensin-converting enzyme gene does not contribute to genetic susceptibility to systemic sclerosis in European Caucasians. AB - OBJECTIVE: To determine whether angiotensin-converting enzyme (ACE) polymorphisms including I/D and 2 single-nucleotide polymorphisms (SNP) affect susceptibility to systemic sclerosis (SSc) in a large French Caucasian population. METHODS: A case-control study was performed in 494 patients with SSc and 280 healthy controls for I/D polymorphism. Two supplementary exonic SNP of ACE gene (rs4309, rs4362) were genotyped in 659 patients with SSc and 511 matched healthy controls. Among the whole SSc population, 453 (67%) patients with SSc had the limited cutaneous subtype, 47 (7%) had precapillary pulmonary arterial hypertension, 209 (32%) had digital ulcers, and 10 (1.5%) had renal crisis. A combined analysis of the available results for ACE I/D genotypes in Caucasians was also performed. RESULTS: There was no association between the 3 polymorphic markers and SSc for allelic and genotype frequencies. No association was observed for the different vascular subsets of the disease. Haplotype analyses did not detect any association. The lack of association for ACE I/D was confirmed by the combined analysis. CONCLUSION: These results in a large cohort of European Caucasian patients with SSc do not support that the ACE gene is implicated in the pathogenesis of SSc and its vascular damage. PMID- 19132787 TI - Apolipoprotein H promoter polymorphisms in relation to lupus and lupus-related phenotypes. AB - OBJECTIVE: Sequence variation in gene promoters is often associated with disease risk. We tested the hypothesis that common promoter variation in the APOH gene (encoding for ss(2)-glycoprotein I) is associated with systemic lupus erythematosus (SLE) risk and SLE-related clinical phenotypes in a Caucasian cohort. METHODS: We used a case-control design and genotyped 345 women with SLE and 454 healthy control women for 8 APOH promoter single-nucleotide polymorphisms (SNP; -1284C>G, -1219G>A, -1190G>C, -759A>G, -700C>A, -643T>C, -38G>A, and 32C>A).Association analyses were performed on single SNP and haplotypes. Haplotype analyses were performed using EH (Estimate Haplotype-frequencies) and Haploview programs. In vitro reporter gene assay was performed in COS-1 cells. Electrophoretic mobility shift assay (EMSA) was performed using HepG2 nuclear cells. RESULTS: Overall haplotype distribution of the APOH promoter SNP was significantly different between cases and controls (p = 0.009). The -643C allele was found to be protective against carotid plaque formation (adjusted OR 0.37, p = 0.013) among patients with SLE. The -643C allele was associated with a ~2-fold decrease in promoter activity as compared to wild-type -643T allele (mean +/- standard deviation: 3.94 +/- 0.05 vs 6.99 +/- 0.68, p = 0.016). EMSA showed that the -643T>C SNP harbors a binding site for a nuclear factor. The -1219G>A SNP showed a significant association with the risk of lupus nephritis (age-adjusted OR 0.36, p = 0.016). CONCLUSION: Our data indicate that APOH promoter variants may be involved in the etiology of SLE, especially the risk for autoimmune mediated cardiovascular disease. PMID- 19132788 TI - Adipokines and systemic lupus erythematosus: relationship with metabolic syndrome and cardiovascular disease risk factors. AB - OBJECTIVE: To study concentrations of adipokines in patients with systemic lupus erythematosus (SLE) and the relationship among adipokines, the metabolic syndrome (MeS), and cardiovascular disease (CVD) risk factors. METHODS: We enrolled 50 SLE patients and 26 controls, all women. Leptin, resistin, visfatin, and adiponectin were measured by commercial ELISA kits. RESULTS: MeS prevalence was increased among subjects with SLE. Leptin levels were higher in patients with SLE than controls. Among SLE patients, independent determinants of leptin were insulin levels (p < 0.0001), triglycerides (p = 0.03), body mass index (p = 0.02), corticosteroid dosage (p = 0.02), and SLE Disease Activity Index (p = 0.005). Other adipokines did not differ between SLE patients and controls. CONCLUSION: Leptin was increased in SLE patients and could play a role in SLE-related cardiovascular diseases. PMID- 19132789 TI - A safety assessment of tumor necrosis factor antagonists during pregnancy: a review of the Food and Drug Administration database. AB - OBJECTIVE: To present any congenital anomalies with respect to tumor necrosis factor (TNF) antagonists reported to the US Food and Drug Administration (FDA) to determine if there are common findings. METHODS: A review of the FDA database of reported adverse events with etanercept, infliximab, and adalimumab from 1999 through December of 2005 was performed. Key words for congenital anomalies were employed as search tools. Duplicate reports were eliminated. Any concomitant medicines were recorded. RESULTS: Our review of > 120,000 adverse events revealed a total of 61 congenital anomalies in 41 children born to mothers taking a TNF antagonist. Of these mothers, 22 took etanercept and 19 took infliximab. There were no reports in women taking adalimumab. The most common reported congenital anomaly was some form of heart defect. Twenty-four of the 41 (59%) children had one or more congenital anomalies that are part of vertebral abnormalities, anal atresia, cardiac defect, tracheoesophageal, renal, and limp abnormalities (VACTERL) association. There were 34 specific types of congenital anomalies in total, and 19 (56%) of those are part of the VACTERL spectrum. Nine of these 19 (47%) types of VACTERL anomalies were observed statistically significantly more than historical controls (p < 0.01); in 4 of these 9 the p value was < or = 0.0001. Thirteen (32%) of the children had more than one congenital anomaly; 7 of these 13 children had 2 defects that are part of the VACTERL spectrum. However, only 1 child was diagnosed with VACTERL. In 24/41 cases (59%) the mother was taking no other concomitant medications. CONCLUSION: A seemingly high number of congenital anomalies that are part of the VACTERL spectrum have been reported. These congenital anomalies are occurring at a rate higher than historical controls. This commonality raises concerns of a possible causative effect of the TNF antagonists. PMID- 19132790 TI - Symptoms of depression predict the trajectory of pain among patients with early inflammatory arthritis: a path analysis approach to assessing change. AB - OBJECTIVE: To assess the longitudinal relationships, including directionality, among chronic pain, symptoms of depression, and disease activity in patients with early inflammatory arthritis (EIA). METHODS: One hundred eighty patients with EIA completed an examination, including swollen joint count, and were administered the Center for Epidemiological Studies Depression Scale (CES-D) and the McGill Pain Questionnaire (MPQ) at 2 timepoints 6 months apart. Cross-lagged panel path analysis was used to simultaneously assess concurrent and longitudinal relationships among pain, symptoms of depression, and number of swollen joints. RESULTS: Pain, symptoms of depression, and number of swollen joints decreased over time (p < 0.001) and were prospectively linked to pain, symptoms of depression, and number of swollen joints, respectively, at 6 months. Symptoms of depression and pain were correlated with each other at baseline (0.47) and at 6 month followup assessments (0.28). Baseline symptoms of depression significantly predicted pain symptoms at 6 months (standardized regression coefficient = 0.28, p = 0.001), whereas pain and disease activity did not predict the course of any other variable after controlling for baseline values. CONCLUSION: Symptoms of depression predicted the trajectory of pain from baseline to 6 months. In addition, there were reciprocal/bidirectional associations between pain and symptoms of depression over time. More research is needed to better understand the relationship between pain and depressive symptoms and how to best manage patients with EIA who have high levels of both. PMID- 19132791 TI - The minimally important difference for the health assessment questionnaire in rheumatoid arthritis clinical practice is smaller than in randomized controlled trials. AB - OBJECTIVE: Patient-reported outcomes are used in clinical practice and trials. We studied a large clinical practice to determine the minimally important difference (MID) estimates for (1) the Health Assessment Questionnaire-Damage Index (HAQ DI): improvement and worsening using patient global assessment anchor; and (2) pain using a patient-reported pain anchor. METHODS: Patients with rheumatoid arthritis (RA; N = 225) had clinic visits at 2 timepoints within 1 year, completed the HAQ-DI and pain visual analog scale (VAS; 0-100 mm), and answered the question, "How would you describe your overall status/overall pain since the last visit?", as much worsened, somewhat worsened, the same, somewhat improved, or much improved. If rated as somewhat improved or worsened, they were defined as the minimally changed subgroups. RESULTS: Eighty-three percent were women, mean age 60 years, with disease duration 11.7 +/- 10.7 years. The baseline HAQ-DI was 0.97 +/- SD 0.76, and at followup 1.0 +/- 0.77 (mean change +0.03 +/- 0.40). The baseline pain VAS was 42.3 +/- 28.8, and at followup 38.5 +/- 27.9 (mean change 2.8 +/- 25.9). The mean (SD) HAQ-DI change score was -0.09 (0.42) for somewhat improved and 0.15 (0.33) for somewhat worsened. The HAQ-DI change for somewhat/much better was -0.20 +/- 0.52, and for somewhat/much worse +0.21 +/- 0.33. For pain, somewhat improved changed by -11.9 mm on the VAS, and somewhat worsened by 6.8 mm. Estimates for HAQ-DI and pain were larger than the for no change group, 0.03 (0.32) and -3.2 (20.9). CONCLUSION: The MID for HAQ-DI in clinical practice is smaller than it is in trials. This may have implications for observational studies and clinical care. PMID- 19132792 TI - Cartilage and bone biomarkers in rheumatoid arthritis: prediction of 10-year radiographic progression. AB - OBJECTIVE: As current predictors of joint destruction have low specificity, serological biomarkers reflecting bone and cartilage destruction have been proposed as tools in assessing prognosis of rheumatoid arthritis (RA). We examined whether serum concentrations of a panel of biomarkers could predict radiographic progression in patients with RA. METHODS: A cohort of 238 patients with RA was followed longitudinally for 10 years with collection of clinical data and serum samples. These analyses focus on the 136 patients with radiographs of the hands available at baseline and at 5 and/or 10 years. Radiographs were scored according to the van der Heijde-modified Sharp score (SHS). Baseline sera were analyzed for receptor activator of nuclear factor-kappaB ligand (RANKL), osteoprotegerin (OPG), human cartilage glycoprotein-39 (YKL-40), C2C, collagen cross-linked C-telopeptide (CTX-I), and cartilage oligomeric matrix protein (COMP). Multivariate linear and logistic regression analyses were used to identify predictors of radiographic progression. RESULTS: Baseline CTX-I levels were higher in progressors [0.41 ng/ml (interquartile range 0.31-0.75)] than in nonprogressors [0.32 ng/ml (IQR 0.21-0.49)], and were independently associated with 10-year change in radiographic damage score [ss = 16.4 (IQR 5.7-27.1)]. We found no association between radiographic progression and baseline serum levels of RANKL, OPG, C2C, YKL-40, or COMP. CONCLUSION: This longterm followup study of patients with RA indicates a relationship between elevated CTX-I levels in serum and subsequent joint destruction. This association was, however, weak, and our study does not support that serum CTX-I or any of the other tested biomarkers will serve as more useful prognostic markers than current predictors such as anti cyclic citrullinated peptide, radiographic damage early in the disease course, and signs of inflammation. PMID- 19132793 TI - Rheumatoid atlantoaxial subluxation can be prevented by intensive use of traditional disease modifying antirheumatic drugs. AB - OBJECTIVE: To evaluate the 5-year incidence of cervical spine disorders in patients with early rheumatoid arthritis (RA) treated by 2 different disease modifying antirheumatic drug (DMARD) strategies. METHODS: In a national, multicenter, prospective FIN-RACo-trial, a cohort of 199 patients with early, clinically active RA was randomly assigned to treatment with a combination of 3 DMARD and prednisolone (Combi group) or with a single DMARD (Single group) with or without prednisolone, aiming to induce remission. After 2 years, the DMARD therapy was unrestricted. Lateral view cervical spine radiographs during full flexion and extension were taken at the 5-year followup visits. The presence of anterior atlantoaxial subluxation (aAAS), atlantoaxial impaction (AAI), and subaxial subluxation (SAS) was assessed in the 149 patients with radiographs available (80 Single and 69 Combi). RESULTS: At the 5-year visits, aAAS, AAI, and SAS were found in 13 (9%), 6 (4%), and 9 (6%) patients, respectively. The corresponding Single/Combi group ratios were 11/2, 5/1, and 5/4. Of the baseline data, only poor physical function [Health Assessment Questionnaire (HAQ); p = 0.024] and Single treatment strategy (p = 0.019) were significantly associated with aAAS. Worse HAQ scores and Disease Activity Score 28 values were found in patients who developed aAAS during the 5-year followup. CONCLUSION: RA patients with sustained clinical disease activity and poor HAQ are at increased risk of developing aAAS. The development of aAAS during the first 5 years of RA was rare among the patients treated with a combination of DMARD for at least 2 years from the diagnosis. Intensive treatment with traditional DMARD prevents or retards the development of aAAS in patients with recent-onset RA. PMID- 19132794 TI - Rheumatoid arthritis and fibromyalgia: a frequent unrelated association complicating disease management. AB - OBJECTIVE: To assess the value of the 28-joint Disease Activity Score (DAS28) in evaluating disease activity in rheumatoid arthritis (RA) associated with fibromyalgia (FM). In this situation, because of the weight of the subjective measures included in the DAS28 equation, the patient's status may be overestimated, leading to inappropriate treatment. We analyze the relationship between RA and FM and discuss whether the association is random or a marker of poor prognosis. METHODS: A questionnaire, developed when biologic therapies were introduced, was administered and the results analyzed in a consecutive, female outpatient population including 105 patients with RA, 49 with RA and FM (RAF), and 28 with FM. Psychosocial characteristics, disease presentation, and radiographic joint destruction evaluation were compared in the 3 populations. RESULTS: The presentation of RA was the same in patients with RA and RAF, but the 2 populations differed by socioprofessional characteristics, significantly higher disease activity in patients with RAF, and significantly more severe joint destruction in patients with RA. The RAF group was similar to the FM control population in socioprofessional and some physical characteristics. Regression analysis using the DAS28 measures differed significantly in the weight allowed to 28-joint counts for pain and swelling, but the constant factor was higher in patients with RAF. CONCLUSION: DAS28 overestimated objective RA severity in patients who also had FM. The association between RA and FM does not appear to be a marker of worse prognosis, but rather a fortuitous association between the 2 diseases and one that may afford these patients some protection against joint destruction. PMID- 19132795 TI - Interaction of brain and intracardiac levels of rhythmogenesis hierarchical system at heart rhythm formation. AB - A single-stage bilateral conduction blockade of the vagus nerves (functional denervation) by constant anodal current was carried out in 13 dogs which are under anesthesia and 3-5 days after operation in chronic experiments. In anesthetized animals, "functional denervation" led to acceleration of the heart rhythm from 102.4+/-3.2 bmp to 123.8+/-4.4 bmp. In chronic dogs "functional denervation" led to transient stoppage of the heart--a preautomatic pause with duration of 2.7+/-0.2 sec. The heartbeats recommenced with the frequency of 89.0+/-3.4 bmp versus an initial rhythm of 118+/-1.5 bpm, i.e., a rhythm deceleration took place. We conclude that in a whole organism the heart rhythm pacemaker is determined by a brain level of the hierarchical system of rhythmogenesis, while the sinoatrial node plays the role of a latent pacemaker. PMID- 19132796 TI - Meg evaluation of epileptic activity in the time and frequency domain. AB - We investigated the localization of current sources in the time and frequency domain from spontaneous MEG data recorded from nine epileptic patients (six females; three males) randomly selected, who had a mean age of 41 years old (range of 17-78 years old), with different types of epilepsy. The MEG data were recorded in a magnetically shielded room with a whole-head 122 channel biomagnetometer. For each MEG spike, we calculated the single Equivalent Current Dipole (ECD) sources at the initial spike peaks with a spherical model. MRI and EEG findings were available in patients' records. Prominent low frequencies can be seen in the majority of channels. For each patient there was an increase of the frequency range after the ECD in comparison with the frequency range before the ECD, in the whole study group due to epileptic discharge which is statistically significant (p=0.02). There was also a statistical significant difference in the increase of the frequency range in four patients with pathologic MRI (p=0.05), in five patients with normal MRI (p=0.02), in five patients with a high incidence of seizures (p=0.04) and in four patients with onset<10 years (p=0.04). The MEG analysis of neuromagnetic data gives information about the modification of the frequency range in the epileptic brains. PMID- 19132797 TI - A model of the cerebellar sensory--motor control applied to fast human forearm movements. AB - To address the problem of how the cerebellum processes the premotor orders that control fast movements of the forearm, a model of the cerebellar control is proposed: a cybernetic circuit composed of a model of the cerebellar premotor pathways driving a biomechanical model of the human forearm. Experiments consist of recording electromyographic (EMG) activities and cinematic variables of the human forearm during fast, single joint, point-to-point movements performed in horizontal and vertical directions with and without mass. The biomechanical model of the forearm is first validated by comparing actual movements and movements simulated by using, as inputs to this model, the synthesized EMG signals and of real EMG activities recorded during the experiments. Then the entire control model is validated by comparing actual movements to the desired ones simulated by the model of the cerebellar pathways whose inputs are velocity signals with Gaussian time-courses. The results show that approximate inverse functions can be computed by means of inner models of direct functions placed in feedback loops, and suggest that the orientation of any member segment with respect to gravity is computed as a cinematic variable in the Central Nervous System (CNS). PMID- 19132798 TI - Neural mechanisms of auditory discrimination of long-duration tonal patterns: a neural modeling and fMRI study. AB - Language perception comprises mechanisms of perception and discrimination of auditory stimuli. An important component of auditory perception and discrimination concerns auditory objects. Many interesting auditory objects in our environment are of relatively long duration; however, the temporal window of integration of auditory cortex neurons processing these objects is very limited. Thus, it is necessary to make active use of short-term memory in order to construct and temporarily store long-duration objects. We sought to understand the mechanisms by which the brain manipulates long-duration tonal patterns, temporarily stores the segments of those patterns, and integrates them into an auditory object. We extended a previously constructed model of auditory recognition of short-duration tonal patterns by expanding the prefrontal cortically-based short-term memory module of the previous model into a memory buffer with multiple short-term memory submodules and by adding a gating module. The gating module distributes the segments of the input pattern to separate locations of the extended prefrontal cortex in an orderly fashion, allowing a subsequent comparison of the stored segments against the segments of a second pattern. In addition to simulating behavioral data and electrical activity of neurons, our model also produces simulations of the blood oxygen level dependent (BOLD) signal as obtained in fMRI studies. The results of these simulations provided us with predictions that we tested in an fMRI experiment with normal volunteers. This fMRI experiment used the same task and similar stimuli to that of the model. We compared simulated data with experimental values. We found that two brain areas, the right precentral gyrus and the left medial frontal gyrus, correlated well with our simulations of the memory gating module. Other fMRI studies of auditory perception and discrimination have also found correlation of fMRI activation of those areas with similar tasks and thus provide further support to our findings. PMID- 19132799 TI - Neural mechanisms of ERP change: combining insights from electrophysiology and mathematical modeling. AB - Using a standardized database of EEG data, recorded during the habituation and oddball paradigms, changes in the auditory event-related potential (ERP) are demonstrated on the time scale of seconds and minutes. Based on previous research and a mathematical model of neural activity, neural mechanisms that could account for these changes are proposed. When the stimulus tones are not relevant to a task, N100 magnitude decreases substantially for the first repetition of a stimulus pattern and increases in response to a variant tone. It is argued these short-term changes are consistent with the hypothesis that there is a refractory period in the neural elements underlying the ERP. In the oddball paradigm, when the stimulus tones are task-relevant, the magnitudes of both N100 and P200 for backgrounds decrease over the entire six-minute recording session. It is argued that these changes are mediated by a decreasing arousal level, and consistent with this, a subject's electrodermal activity (EDA) is shown to reduce over the recording session. By fitting ERPs generated by a biophysical model of neural activity, it is shown that the changes in the background ERPs over the recording session can be reproduced by changing the strength of connections between populations of cortical neurons. For ERPs elicited by infrequent stimuli, there is no corresponding trend in the magnitudes of N100 or P300 components. The effects of stimuli serial order on ERPs are also assessed, showing that the N100 for background ERPs and the N100 and P300 for target ERPs increases as the probability, and expectancy, of receiving a task relevant stimulus increases. Cortical neuromodulation by acetylcholine (ACh) is proposed as a candidate mechanism to mediate the ERP changes associated with attention and arousal. PMID- 19132800 TI - The self and its awareness: genesis of psychoses. AB - Agnosias demonstrate the broadest spectrum of pathology of consciousness in neurology and psychiatry. Agnosias wipe off the definite functions from the brain's activity precisely and completely, allowing the consciousness to be investigated in itself. Thorough investigations of confabulatory manifestations disclose the rationale for the development of pathological functions and point out that there is a remedial (reconstructive) sense behind the senselessness of a mental state. Pathology seems to be accompanied by involuntary reparation on the part of the brain. Investigations of the conscious activity in agnosias show a separate, passive and real brain system of consciousness without pathology and genetic regulations. Neurological symbolic agnosias represent a cerebral and notional character and therefore constitute a link between neurological and psychiatric agnosias, which proves the existence of the same mechanism of development underlying agnosic syndromes, both in psychiatry and neurology. The work proves dual form of human self (somatic and notional) that constitute oneness, when they are dissociated, agnosia is caused. PMID- 19132801 TI - Vision disorders and phosphodiesterase type 5 inhibitors: a review of the evidence to date. AB - Phosphodiesterase type 5 (PDE-5) inhibitors (sildenafil, vardenafil and tadalafil) have been in widespread use for the safe and effective treatment of erectile dysfunction (ED) for nearly a decade. During that time, a relatively small number of patients have experienced adverse visual events, including nonarteritic anterior ischaemic optic neuropathy (NAION). In this article, post marketing reports of adverse visual events along with other relevant literature on ocular safety related to PDE-5 inhibitor use are reviewed. Although a relatively small number of cases have been reported with a possible temporal association with PDE-5 inhibitor use, it has not been possible to conclude whether these events are coincidental or whether they are associated with effects of PDE-5 inhibitors on ocular circulation or on other structures of the eye. A careful review of pooled data from clinical trials for all three PDE-5 inhibitors, which contain well documented information about the dose and duration of exposure to the drug for a large number of patients, yields no evidence for an increased risk of NAION or other adverse ocular events associated with PDE-5 inhibitor use. However, the inherent limitations in interpreting results from clinical trials and potentially incomplete information from post-marketing surveillance preclude a definitive declaration that ocular safety will not be a concern for some patients with ED and co-morbid disease states. Despite the absence of a proven link between PDE-5 use and serious ocular disorders, physicians should continue to advise patients to stop use of a PDE-5 inhibitor and seek immediate medical attention in the event of a sudden loss of vision as a safety measure. PMID- 19132802 TI - Determinants of under-reporting of adverse drug reactions: a systematic review. AB - A voluntary reporting system of adverse drug reactions (ADRs) is fundamental to drug safety surveillance but under-reporting is its major limitation. This bibliographic review sought to assess the influence of personal and professional characteristics on ADR reporting and to identify knowledge and attitudes associated with ADR reporting. A systematic review was conducted using the MEDLINE and EMBASE databases. We included papers that were published in English, French and Spanish, and covered a study population made up of health professionals. In each case, the following data were extracted: study population; workplace; study type; sample size; type of questionnaire; type of scale for measuring knowledge; response rate; personal and professional factors; and knowledge and attitudes (based on Inman's 'seven deadly sins') associated with reporting. Based on a search of computerized databases, we identified a total of 657 papers in MEDLINE and 973 in EMBASE. In all, the review covered 45 papers that fulfilled the inclusion criteria. Medical specialty was the professional characteristic most closely associated with under-reporting in 76% of studies involving physicians. Other factors associated with under-reporting were ignorance (only severe ADRs need to be reported) in 95%; diffidence (fear of appearing ridiculous for reporting merely suspected ADRs) in 72%; lethargy (an amalgam of procrastination, lack of interest or time to find a report card, and other excuses) in 77%; indifference (the one case that an individual doctor might see could not contribute to medical knowledge) and insecurity (it is nearly impossible to determine whether or not a drug is responsible for a particular adverse reaction) in 67%; and complacency (only safe drugs are allowed on the market) in 47% of studies. While personal and professional factors display a weak influence, the knowledge and attitudes of health professionals appear to be strongly related with reporting in a high proportion of studies. This result may have important implications in terms of public health, if knowledge and attitudes are viewed as potentially modifiable factors. PMID- 19132803 TI - Safety of rupatadine administered over a period of 1 year in the treatment of persistent allergic rhinitis: a multicentre, open-label study in Spain. AB - BACKGROUND: Rupatadine (Rupafin), a novel antihistamine approved recently in Europe for the treatment of allergic rhinitis (AR) and chronic idiopathic urticaria in patients aged>or=12 years, has been shown to be highly efficacious, and as safe and well tolerated as other commonly employed antihistamines in the treatment of allergic disease. There are, however, few data on the long-term safety of these antihistamines derived in accordance with the clinical safety recommendations of the European Agency for the Evaluation of Medicinal Products (EMEA) and the International Conference on Harmonisation (ICH) of Technical Requirements for Registration of Pharmaceuticals for Human Use Guideline. OBJECTIVE: To assess the safety and tolerability of treatment with rupatadine 10 mg/day for 12 months in subjects with persistent AR (PER). METHODS: A multicentre, open-label, phase IV study in patients recruited from 33 centres in Spain, from September 2002 to November 2005. The study enrolled 324 male and female patients (aged 12-70 years) with a medical history of PER for at least 12 months and a documented positive skin-prick test to an appropriate allergen. On 4 of the 7 days prior to start of treatment, the patients were required to have a minimum total nasal symptom score (TNSS [for sneezing, rhinorrhoea, nasal obstruction/congestion and nasal itching]) of >or=5. Of the 324 eligible patients starting treatment, 120 needed to be treated for more than 6 months and were followed up until the end of 12 months. All patients received rupatadine 10 mg/day and were allowed to continue their normal concomitant medication for all conditions, other than rhinitis, for up to 6 or 12 months. Safety was assessed by means of adverse events (AEs) reported by patients or detected by investigators, scheduled centralized ECG with special attention to Bazzet corrected QT interval (QTcB) and standard laboratory investigations. RESULTS: Assessment of treatment compliance rates indicated 90% and 83% of patients to be compliant during the 1-6 months and 1-12 months treatment periods, respectively, with compliance rates>80% being associated with the majority of the study population reporting at least one AE. Overall, 74.1% and 65.8% of the patients reported at least one AE during the 1-6 months and 1-12 months treatment periods, respectively, compared with 20.4% and 10.8% of patients reporting at least one treatment-related AE during these periods. Disorders of the nervous system and respiratory thoracic and mediastinal system, in particular headache, somnolence and catarrh, were the three most common AEs reported by >5% of the patients during both treatment periods. Detailed ECG assessments demonstrated no clinically relevant abnormal ECG findings, nor any QTcB increases >60 msec or QTcB values>470 msec for any patient at any time during treatment. Serious AEs were reported in seven patients, of whom six were considered as unlikely to be related to rupatadine treatment, whereas one involving increased blood enzyme levels was considered as possibly related to rupatadine treatment. CONCLUSION: This study confirmed the good long term safety and tolerability of rupatadine at the therapeutic dose of 10 mg/day in patients with PER. PMID- 19132804 TI - Use and risk management of carvedilol for the treatment of heart failure in the community in England: results from a modified prescription-event monitoring study. AB - BACKGROUND: In the UK, the licence for carvedilol was extended in 1998 to include symptomatic heart failure (New York Heart Association [NYHA] class II and III heart failure) with the recommendation that initiation and up-titration should be under the supervision of a hospital physician. A post-marketing surveillance study was conducted to address the UK regulatory authority's request for monitoring the use and safety of carvedilol prescribed for heart failure in clinical practice. AIM: To investigate adherence to risk management recommendations for the use of carvedilol for heart failure, monitor how patients' subsequent care was managed and collect event data to evaluate the safety profile of carvedilol used for the treatment of heart failure. METHODS: An observational cohort study using a modified prescription-event monitoring technique identified patients from dispensed primary care prescriptions in England (August 1999 to June 2001). An eligibility questionnaire was used to identify patients who had been prescribed carvedilol for heart failure for the first time after 31 July 1999. Up to three follow-up questionnaires were sent to the prescribers of eligible patients, requesting demographic information, dosage, supervision of treatment, status of cardiac failure and event information. RESULTS: 2311 patients met the eligibility criteria. For 1666 patients, one or more valid follow-up questionnaires were returned: 68.5% were male; male median age 66 years; female median age 72 years; the observation period was up to 3 years. Hospital physicians supervised initiation of treatment and first up titration in 85.6% and 61.4% of patients, respectively. 49.2% of patients were prescribed the recommended starting dosage of carvedilol (6.25 mg/day). Approximately 25% of patients started on a lower dose than recommended, and the same proportion were prescribed a higher dose. NYHA status of cardiac failure between starting treatment and the third questionnaire improved for 39.5% of patients, deteriorated for 10.9%, and 11.7% of those for whom NYHA status was given died. Adverse drug reactions (ADRs) were reported for 2.4% of patients; the most commonly reported ADR was malaise/lassitude. Overall, 27.1% of patients stopped taking carvedilol. None of the 163 deaths were attributed to carvedilol. CONCLUSIONS: Regulatory guidelines for the use and risk management of carvedilol in heart failure were mostly followed, and most patients appeared to benefit from treatment with carvedilol for heart failure. Malaise/lassitude was the main reason for discontinuing treatment. Further investigations may be warranted to examine the prescribing of carvedilol at lower than recommended doses. PMID- 19132806 TI - Platelet measurements versus discharge diagnoses for identification of patients with potential drug-induced thrombocytopenia: a cross-sectional study in the Netherlands. AB - BACKGROUND: In pharmacoepidemiological studies on the risk of drug-induced blood dyscrasias, including drug-induced thrombocytopenia (DIT), hospital discharge diagnoses have been used to identify potential cases. One of the possible limitations of discharge diagnoses is that due to incomplete registration not all potential cases are identified, which may limit statistical power. Clinical laboratory data have been suggested as a data type that is potentially more sensitive for identifying potential cases of adverse drug reactions than discharge diagnoses. OBJECTIVE: To compare the number of patients with potential DIT that could be identified by using platelet measurements with the number of patients with potential DIT that could be identified by using discharge diagnoses for thrombocytopenia within a population of hospitalized patients. METHODS: The study population of this cross-sectional study comprised all patients admitted to the University Medical Center Utrecht in 2004 and 2005, as captured within the Utrecht Patient Oriented Database (UPOD). The ratio of the number of patients with potential DIT based on platelet measurements (>or=1 platelet count below 100x10(9)/L without alternative diagnoses for DIT) to the number of patients with potential DIT based on discharge diagnoses for thrombocytopenia (International Classification of Diseases, 9th Revision, Clinical Modification [ICD-9-CM] codes 287.3-287.5 without alternative diagnoses for DIT) was determined. RESULTS: Within the study period there were 56,411 hospitalizations. 2817 patients (5.0%) had >or=1 platelet count below 100x10(9)/L. In 96.3% of these patients, alternative diagnoses for DIT were present, resulting in 103 (0.2%) patients with potential DIT based on platelet measurements. There were 74 patients (0.1%) with a discharge diagnosis for thrombocytopenia. In 81.1% of these patients, alternative diagnoses for DIT were present, resulting in 14 (0.02%) patients with potential DIT based on discharge diagnoses. This resulted in a ratio of the number of patients with potential DIT based on platelet measurements to the number of patients with potential DIT based on discharge diagnoses for thrombocytopenia of seven. CONCLUSION: This study showed that the use of platelet measurements is a more sensitive approach to the identification of patients with potential DIT than the use of discharge diagnoses for thrombocytopenia. PMID- 19132807 TI - An intervention to improve spontaneous adverse drug reaction reporting by hospital physicians: a time series analysis in Spain. AB - BACKGROUND: Spontaneous reporting of adverse drug reactions (ADRs) in hospitals is scarce and several obstacles to such reporting have been identified previously. OBJECTIVE: To assess the effectiveness of a multifaceted intervention based on healthcare management agreements for improving spontaneous reporting of ADRs by physicians in a hospital setting. METHODS: In 2003, the spontaneous reporting of ADRs was included as one of the objectives of hospital physicians at the Vall d'Hebron Hospital, Barcelona, Spain, within the context of management agreements between clinical services and hospital managers. A continuous intervention related to these management agreements, including periodic educational meetings and economic incentives, was then initiated. We carried out an ecological time series analysis and assessed the change in the total number of spontaneous reports of ADRs, and the number of serious ADRs, unexpected ADRs, and ADRs associated with new drugs between a period previous to the intervention (from 1998 to 2002) and the period during the intervention (from 2003 to 2005). A time series analysis with ARIMA (Auto-Regressive Integrated Moving Average) models was performed. RESULTS: The median number of reported ADRs per year increased from 40 (range 23-55) in the first period to 224 (range 98-248) in the second period. In the first period, the monthly number of reported ADRs was stable (3.47 per month; 95% CI 1.90, 5.03), but in the second period the number increased progressively (increase of 0.74 per month; 95% CI 0.62, 0.86). In the second period, the proportion of reported serious ADRs increased nearly 2-fold (63.1% vs 32.5% in the first period). The absolute number of previously unknown or poorly known ADRs increased 4-fold in the second period (54 vs 13 in the first period). There was also an increase in the absolute number of suspected pharmacological exposures to new drugs (97 vs 28) and in the number of different new drugs suspected of causing ADRs (50 vs 19). CONCLUSION: A continuous intervention based on healthcare management agreements with economic incentives and educational activities is associated with a quantitative and qualitative improvement of spontaneous reporting of ADRs by hospital physicians. PMID- 19132805 TI - Drug-Induced Liver Injury Network (DILIN) prospective study: rationale, design and conduct. AB - BACKGROUND: Drug-induced liver injury (DILI) is an uncommon adverse drug reaction of increasing importance to the medical community, pharmaceutical industry, regulatory agencies and the general public. OBJECTIVES: The Drug-Induced Liver Injury Network (DILIN) was established to advance understanding and research into DILI by initiating a prospective registry of patients with bona fide DILI for future studies of host clinical, genetic, environmental and immunological risk factors. The DILIN was also charged with developing standardized nomenclature, terminology and causality assessment instruments. METHODS: Five clinical sites, a data coordinating centre and senior scientists from the National Institute of Diabetes and Digestive and Kidney Diseases initiated the DILIN prospective study in September 2004. Eligible patients are required to meet minimal laboratory or histological criteria within 6 months of DILI onset and have other competing causes of liver injury excluded. Patients in the general community setting with pre-existing HIV, hepatitis B virus or hepatitis C virus infections and/or abnormal baseline liver biochemistries are eligible for enrollment. In addition, subjects with liver injury due to herbal products are eligible to participate. Control patients without DILI are also to be recruited in the future. RESULTS: All referred subjects undergo an extensive review of available laboratory, pathology and imaging studies. Subjects who meet pre-defined eligibility criteria at the 6-month study visit are followed for 2 years to better define the natural history of chronic DILI. Causality assessment is determined by a panel of three hepatologists who independently assign a causality score ranging from 1 (definite) to 5 (unlikely) as well as a severity score ranging from 1 (mild) to 5 (fatal). During the first 3 years, 367 subjects were enrolled into the DILIN prospective study. CONCLUSION: DILIN is a multicentre research network charged with improving our understanding of the aetiologies, risk factors and outcomes of DILI in the US. The network is meeting the targeted enrollment of ten patients per month and is developing a repository of clinical data and biological samples for future studies of DILI pathogenesis and outcome. PMID- 19132808 TI - Drug-induced thrombocytopenia: an updated systematic review, 2008. PMID- 19132809 TI - Spatial and temporal patterns of periphyton chlorophyll a related to pulp and paper mill discharges in four US receiving streams. AB - Nutrients in pulp and paper mill effluent (PPME) have been implicated in increased periphyton chlorophyll a (chl a) downstream of discharges. These findings are largely based on short-term studies conducted in artificial stream channels or mesocosms and often in oligotrophic systems, and it is unclear if long-term chl a patterns in higher-nutrient systems would show similar response. We conducted a long-term study of 4 receiving waters (Codorus Creek, Pennsylvania; the Leaf River, Mississippi; and the McKenzie and Willamette rivers, Oregon) in which periphyton samples and associated data on water quality (nitrogen and phosphorus concentrations, pH, color, and conductivity) and 2 physical habitat variables (depth and current velocity) were collected over an 8 y period from multiple sites upstream and downstream of PPME discharges. Study streams represented different ecoregions, warm- and coldwater systems, gradients of in-stream effluent concentration (<1-33%), and mill process types. General Linear Models examining the main and interaction effects of site, season, and year on periphyton chl a for each of the 4 streams showed periphyton chl a downstream of the PPME discharge in Codorus Creek and the McKenzie River was greater at some, but not all upstream sites, suggesting these differences may be due to factors other than PPME. Mean periphyton chl a ranged from <1 to 285 mg/m2 across streams, with relatively consistent site patterns across seasons and years. Overall, chl a in the spring and summer was greater than in the fall in Codorus Creek and on sand substrates in the Leaf River, with overall differences across years seen on rare occasions in the Leaf and Willamette rivers. Regression models examining environmental-chl a relationships explained 45.4% and 30.2% of variation in periphyton chl a in the McKenzie River and Codorus Creek, respectively, and <10% in the Leaf and Willamette rivers. Physical variables (stream depth and current velocity) were the most important model variables in the McKenzie River, while total nitrogen and color were of greatest importance in Codorus Creek. The findings of this study demonstrate the inherent variability of chl a standing crops, highlight the complexity of lotic periphyton communities, and reiterate the importance of long-term, multi-season studies in elucidating spatial and temporal patterns. PMID- 19132810 TI - Positive correlation between hemosporidian parasitemia and likelihood of PCR detection in co-infected birds. AB - As shown previously, the nested PCR method of detection of avian blood parasites, commonly referred to as Waldenstrom's method, sometimes amplifies only 1 parasite species of the several that may be present in the same bird, and not always the one with the highest parasitemia. This result raises questions regarding the use of the molecular method for the identification of bird parasites. Additionally, it is unclear whether the amplified parasite, among the several present in the same host, reflects the intensity of infection. However, I performed statistical analyses on a dataset in which there were multiply infected birds and showed that the parasites with the highest parasitemia are the most likely to be amplified. Such a positive correlation between the likelihood of amplification and the parasitemias of the different blood parasites supports the use of Waldenstrom's method for the comparison of the parasite content of groups of birds exposed to the same parasites. PMID- 19132811 TI - An enriched stable-isotope approach to determine the gill-zinc binding properties of juvenile rainbow trout (Oncorhynchus mykiss) during acute zinc exposures in hard and soft waters. AB - The objective of the present study was to employ an enriched stable-isotope approach to characterize Zn uptake in the gills of rainbow trout (Oncorhynchus mykiss) during acute Zn exposures in hard water (approximately 140 mg/L as CaCO3) and soft water (approximately 30 mg/L as CaCO3). Juvenile rainbow trout were acclimated to the test hardnesses and then exposed for up to 72 h in static exposures to a range of Zn concentrations in hard water (0-1000 microg/L) and soft water (0-250 microg/L). To facilitate detection of new gill Zn from endogenous gill Zn, the exposure media was significantly enriched with 67Zn stable isotope (89.60% vs. 4.1% natural abundance). Additionally, acute Zn toxicity thresholds (96-h median lethal concentration [LC50]) were determined experimentally through traditional, flow-through toxicity tests in hard water (580 microg/L) and soft water (110 microg/L). Following short-term (< or =3 h) exposures, significant differences in gill accumulation of Zn between hard and soft water treatments were observed at the three common concentrations (75, 150, and 250 microg/L), with soft water gills accumulating more Zn than hard water gills. Short-term gill Zn accumulation at hard and soft water LCS0s (45-min median lethal accumulation) was similar (0.27 and 0.20 microg/g wet wt, respectively). Finally, comparison of experimental gill Zn accumulation, with accumulation predicted by the biotic ligand model, demonstrated that model output reflected short-term (<1 h) experimental gill Zn accumulation and predicted observed differences in accumulation between hard and soft water rainbow trout gills. Our results indicate that measurable differences exist in short-term gill Zn accumulation following acclimation and exposure in different water hardnesses and that short-term Zn accumulation appears to be predictive of Zn acute toxicity thresholds (96-h LC50s). PMID- 19132812 TI - Chronic toxicity of five structurally diverse demethylase-inhibiting fungicides to the crustacean Daphnia magna: a comparative assessment. AB - Demethylase inhibitors (DMIs) are broad-spectrum fungicides that are ubiquitously used in agriculture and medicine. They comprise chemically heterogeneous substances that share a common biochemical target in fungi, the inhibition of a specific step in sterol biosynthesis. Several DMIs are suspected to disrupt endocrine-mediated processes in a range of organisms and to inhibit ecdysteroid biosynthesis in arthropods. It is unclear, however, whether and, if so, to what extent different DMI fungicides have a similar mode of action in nontarget organisms, which in turn would lead to a common chronic toxicity profile. Therefore, we selected a representative of each of the major DMI classes--the piperazine triforine, the pyrimidine fenarimol, the pyridine pyrifenox, the imidazole prochloraz, and the triazole triadimefon--and comparatively investigated their chronic toxicity to Daphnia magna. No toxicity was detectable up to the limit of solubility of triforine (61 micromol/L). All other DMIs reduced reproductive success by delaying molting and development and by causing severe developmental abnormalities among offspring. Prochloraz was most toxic (median effective concentration [EC50] for fecundity reduction, 0.76 micromol/L), followed by fenarimol (EC50, 1.14 micromol/L), pyrifenox (EC50, 3.15 micromol/L), and triadimefon (EC50, 5.13 micromol/L). Mean effect concentrations for fecundity reduction were related to lipophilicity and followed baseline toxicity. However, triadimefon and fenarimol (but none of the other tested DMIs) caused severe eye malformations among exposed offspring. Affected neonates did survive, but a reduced ecological fitness can be assumed. Offspring exposed to fenarimol in mater matured earlier. The investigated different life-history parameters were affected in a substance-specific manner. These qualitatively different toxicity profiles suggest additional, substance-specific mechanisms of action in D. magna that probably are related to an antiecdysteroid action. PMID- 19132813 TI - Effects of terrestrial pollutants on insect parasitoids. AB - Parasitoids are important organisms in the regulation of insect herbivores in natural, urban, and agricultural ecosystems. The impact of pollutants acting on parasitoids has not been extensively reviewed. This prompted us to propose a falsifiable null hypothesis (pollutants have no effects on parasitoids) and two alternative hypotheses (pollution negatively or positively affects parasitoids) to assess in the available literature the effects of pollutants acting on parasitoids. We found 26 studies examining 39 biological systems that met our criteria for inclusion. Of these studies, 18 of the 39 biological systems (46.2%) supported the null hypothesis while 18 (46.2%) supported the first alternative hypothesis in which pollutants exhibited negative effects on parasitoids. Only a small percentage of the studies (7.6%, 3 of 39) supported the second alternative hypothesis suggesting that pollutants had positive effects on parasitoids. We provide a synthesis of the available data by pollution type, summarize trends for different pollutants, and suggest future areas of research. PMID- 19132814 TI - Ocean acidification: a prognosis and treatment for this eclipsing issue in marine ecotoxicology. PMID- 19132815 TI - Reducing uncertainty in environmental risk assessment (ERA): clearly defining acute and chronic toxicity tests. PMID- 19132816 TI - Exposure to inhaled benzene and toluene shows a paradoxical response in American kestrels. PMID- 19132817 TI - Mercury in the Arctic: are we overlooking the ocean? PMID- 19132818 TI - Utility of biomarkers in fish for environmental monitoring. PMID- 19132820 TI - Toward a holistic and risk-based management of European river basins. AB - The European Union Water Framework Directive (WFD) requires a good chemical and ecological status of European surface waters by 2015. Integrated, risk-based management of river basins is presumed to be an appropriate approach to achieve that goal. The approach of focusing on distinct hazardous substances in surface waters together with investment in best available technology for treatment of industrial and domestic effluents was successful in significantly reducing excessive contamination of several European river basins. The use of the concept of chemical status in the WFD is based on this experience and focuses on chemicals for which there is a general agreement that they should be phased out. However, the chemical status, based primarily on a list of 33 priority substances and 8 priority hazardous substances, considers only a small portion of possible toxicants and does not address all causes of ecotoxicological stress in general. Recommendations for further development of this concept are 1) to focus on river basin-specific toxicants, 2) to regularly update priority lists with a focus on emerging toxicants, 3) to consider state-of-the-art mixture toxicity concepts and bioavailability to link chemical and ecological status, and 4) to add a short list of priority effects and to develop environmental quality standards for these effects. The ecological status reflected by ecological quality ratios is a leading principle of the WFD. While on the European scale the improvement of hydromorphological conditions and control of eutrophication are crucial to achieve a good ecological status, on a local and regional scale managers have to deal with multiple pressures. On this scale, toxic pollution may play an important role. Strategic research is necessary 1) to identify dominant pressures, 2) to predict multistressor effects, 3) to develop stressor- and type specific metrics of pressures, and 4) to better understand the ecology of recovery. The concept of reference conditions to define the ecological status is hard to apply and tends to ignore the fact that ecosystems can be highly dynamic. A better understanding of ecosystem responses to changes as well as early warning systems and concepts sensitive to various stressors to discriminate disturbances from natural variation are required. Because ecosystems are closely interconnected, an integrated monitoring, diagnosis, and stressors-based management of the whole water, sediment, groundwater, soil, and air system is required considering land use and the interaction with a changing climate. Extending this holistic approach beyond a consideration of existing pressures by anticipating on future ones to use and protect the aquatic environment in a sustainable way is one of the big challenges. PMID- 19132821 TI - Toward an integrated assessment of the ecological and chemical status of European river basins. AB - Here, recommendations to improve ecological and chemical status assessments in accordance with the European Water Framework Directive (WFD) are made on the basis of experience gained from the MODELKEY project database, linking existing biological and chemical monitoring data of 3 case study river basins (Elbe, Scheldt, and Llobregat). The data analysis within and across river basins revealed major obstacles to be tackled, including scarcity of matching ecological and chemical monitoring sites for cause-effect relationships as well as a general lack of stressor-specific metrics for single biological quality elements (BQE) to enable a comprehensive risk assessment of all predominant stressors, including toxicity. An example of such a metric, which is recommended for the BQE of benthic macroinvertebrates, is the trait-based species-at-risk index (SPEAR) that correlated well with a respective measure for toxic stress, referred to as toxic units, based on simple mixture toxicity concepts. Surprisingly, the assessment of chemical status of a total of 695 monitoring sites for 2000 to 2004 showed that environmental quality standards (EQSs) were exceeded for at least 1 of the currently 41 priority pollutants (PPs) in 92% to 98% of the cases in all 3 of the river basins, which, according to definition, indicates potential effects on ecological status. A comparison of compliance with EQSs for 41 PPs with a respective effect threshold (derived for benthic macroinvertebrates) revealed that the rather conservative concept of chemical status is most likely not protective in all cases. Furthermore, to account for the many other compounds that are detected frequently in European surface waters and that may also have ecotoxicological effects, we introduced a provisional predicted no-effect concentration that is in accordance with the EQS methodology and is suggested to identify potential emerging compounds for which no or insufficient toxicity data exist. In conclusion, this study aims to support the implementation of the WFD by drawing conclusions from the analysis of heterogeneous data sets of various member states and by introducing new tools to move toward an integrated European assessment of ecological and chemical status. PMID- 19132822 TI - Oxidative transformation of tetrachlorophenols and trichlorophenols by manganese dioxide. AB - This study examined the transformation kinetics of three tetrachlorophenols (TeCPs) and three trichlorophenols (TCPs) in the presence of MnO2 under different solution chemistry conditions. The reaction rate measured for each CP decreased as a function of solution pH, and under the same solution chemistry conditions, the measured rates may depend primarily on both the adsorbability at the MnO2 surfaces and the isomeric structures of the CPs. Isomeric effects indicated that chloro substituent on ortho or para positions exhibited faster rates of transformation than on meta positions. Gas chromatography-mass spectrometry analysis with a derivatization method showed that dimers including polychlorinated phenoxyphenols and chlorinated polyhydroxybiphenyl were among the major products for all CPs. Monomeric products were among the major products of 2,4,6-TCP, 2,3,4-TCP, and 2,3,4,6-TeCP, whereas trimeric products also were among the major products of 2,3,4-TCP and 2,4,5-TCP. It appeared that hydroxylation of CPs and formation of dimeric or trimeric products via oxidative coupling were the major reaction mechanisms involved in the oxidation of CPs by MnO2. PMID- 19132823 TI - How focal adhesion size depends on integrin affinity. AB - Understanding how the thermodynamics and kinetics of integrin receptor binding and clustering impact the formation of focal adhesions is important for understanding the mechanisms cells use to sense and respond to physical cues in their environment. Cells on chemically well-defined surfaces were observed to have distributions of focal adhesions shifted toward smaller sizes when presented with higher affinity ligands (Kato, M.; Mrksich, M. Biochemistry 2004, 43, 2699). In this paper, we account for this trend with a simple model in which integrins are treated as particles on a lattice, and their stochastic dynamics are simulated with a kinetic Monte Carlo algorithm. How the trend depends on force coupled growth, membrane fluctuations, and heterogeneity of receptor-ligand interactions is analyzed. Predictions are made for substrates in which the ligands presented can vary in either space or time, so that the model can be validated experimentally. PMID- 19132825 TI - Kit-like 18F-labeling of proteins: synthesis of 4-(di-tert butyl[18F]fluorosilyl)benzenethiol (Si[18F]FA-SH) labeled rat serum albumin for blood pool imaging with PET. AB - Radiosyntheses of 18F-radiopharmaceuticals for positron emission tomography (PET) normally require an extraordinarily high effort of technical equipment and specially trained personnel. We recently reported a novel method for the introduction of fluorine-18 into peptides for PET-imaging based on silicon-18F chemistry (SiFA technique). We herewith introduce the first SiFA-based Kit-like radio-fluorination of a protein (rat serum albumin,RSA) and demonstrate its usefulness for in vivo imaging with microPET in normal rats as well as in a rat heterotropic transplanted heart model. As a labeling agent, we prepared 4-(di tert-butyl[18F]fluorosilyl)benzenethiol (Si[18F]FASH)by simple isotopic exchange in 40-60% radiochemical yield (RCY) and coupled it directly to a Sulfo-SMCC derivatized RSA in an overall RCY of 12% within 20-30 min. The technically simple labeling procedure does not require any elaborated purification procedures and is a straightforward example of a successful application of Si-18F chemistry for in vivo imaging with PET. PMID- 19132827 TI - Quantum mechanical study of the photoisomerizations of bicyclo[4,1,0]hept-2-ene (2-norcarene). AB - The mechanisms of the photochemical isomerization reactions were investigated theoretically using a model system of bicyclo[4,1,0]hept-2-ene (2-norcarene) 1 with the CASSCF (eight-electron/eight-orbital active space) and MP2-CAS methods and the 6-311(d) basis set. The structures of the conical intersections and intersystem crossings, which play a crucial role in such photoisomerization reactions, were obtained. The intermediates and transition structures of the ground-state were also calculated to assist in providing a qualitative explanation of the reaction pathways. Our model investigations suggest that the preferred singlet photoreaction route for 1 is as follows: singlet reactant --> Franck-Condon region --> conical intersection --> intermediate --> transition state --> photoproduct. On the other hand, our theoretical findings indicate that the preferred triplet photoreaction route for 1 is as follows: singlet reactant - > Franck-Condon region --> triplet minimum --> triplet transition state --> intersystem crossing --> intermediate --> singlet transition state --> photoproduct. In particular, the intersystem crossing mechanism found in this work gives a better explanation and supports the available experimental observations. Two kinds of reaction pathways, which can lead to final photoproducts, have been identified: (paths I or III) ring-expansion to form a cycloheptene ring and (paths II or IV) ring-closure to form a methylcyclohexene structure. Both exhibit biradical character. Also, our theoretical investigations strongly indicate that substantial interaction occurs between the cyclopropane moiety and the isolated carbon-carbon double bond in the excited state of (1). PMID- 19132831 TI - Tunable multiresponsive methacrylic acid based inverse opal hydrogels prepared by controlling the synthesis conditions. AB - Methacrylic acid based inverse opal hydrogels (MIOHs) have been prepared by controlling the synthesis conditions, including cross-linker content, solvent content, and water content in solvent mixtures to explore the effect of the synthesis conditions (especially solvent content and mixture) on the response performance. Various response events (pH, solvent, ionic strength, 1,4 phenylenediamine dihydrochloride (PDA) response) have been investigated. For pH, solvent response, the same response behaviors have been observed: both the increased solvent (only ethanol) content and the enhanced water content in solvent will lead to the reduced response level of MIOHs compared to that of the increased cross-linker content. However, two different kinds of response behaviors for ionic strength response have been found by adjusting the synthesis conditions. The kinetics of pH response shows characteristics of a diffusion limited process, and the equilibrium response time is about 20 min, which cannot be reduced by changing the synthesis conditions. The PDA response of MIOHs shows a PDA concentration dependence: with the increase of PDA concentration, the response level increases and the response time decreases. PMID- 19132829 TI - Therapeutic index of gramicidin S is strongly modulated by D-phenylalanine analogues at the beta-turn. AB - Analogues of the cationic antimicrobial peptide gramicidin S (GS), cyclo(Val-Orn Leu-D-Phe-Pro)2, with d-Phe residues replaced by different (restricted mobility, mostly) surrogates have been synthesized and used in SAR studies against several pathogenic bacteria. While all D-Phe substitutions are shown by NMR to preserve the overall beta-sheet conformation, they entail subtle structural alterations that lead to significant modifications in biological activity. In particular, the analogue incorporating D-Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) shows a modest but significant increase in therapeutic index, mostly due to a sharp decrease in hemolytic effect. The fact that NMR data show a shortened distance between the D-Tic aromatic ring and the Orn delta-amino group may help explain the improved antibiotic profile of this analogue. PMID- 19132832 TI - Photoresponsive polymer brushes for hydrophilic patterning. AB - The use of photolabile protecting groups (PGs) as a means to create latent hydrophilic surfaces is presented. Naturally hydrophobic PGs, based on o nitrobenzyl chemistry, are used on polymer side chains, poised for cleavage upon exposure to UV light. Removal of the PGs liberates the hydrophilic polymer, thereby switching the surface wettability from hydrophobic to hydrophilic. This switch can be augmented by increasing the surface roughness. Additionally, this system is also shown to be spatially addressable, a highly desirable property for applications which require specific regions of a surface to switch their wettability. PMID- 19132833 TI - Hydrogen-bonding interactions between water and the one- and two-electron-reduced forms of vitamin K1: applying quinone electrochemistry to determine the moisture content of non-aqueous solvents. AB - Vitamin K(1) (VK(1)) was shown by voltammetry and coulometry to undergo two chemically reversible one-electron reduction processes in acetonitrile (CH(3)CN) containing 0.2 M Bu(4)NPF(6) as the supporting electrolyte. The potential separation between the first and second electron-transfer steps diminished sequentially with the addition of water, so that at a H(2)O concentration of approximately 7 M (approximately 13% v/v) only one process was detected, corresponding to the reversible transfer of two electrons per molecule. The voltammetric behavior was interpreted on the basis of the degree of hydrogen bonding between the reduced forms of VK(1) with water in the solvent. It was found that the potential separation between the first and second processes was especially sensitive to water in the low molar levels (0.001-0.1 M); therefore, by measuring the peak separation as a function of controlled water concentrations (accurately determined by Karl Fischer coulometric titrations) it was possible to prepare calibration curves of peak separation versus water concentration. The calibration procedure is independent of the type of reference electrode and can be used to determine the water content of CH(3)CN between 0.01 and 5 M, by performing a single voltammetric scan in the presence of 1.0 mM VK(1). The voltammetry was also investigated in dichloromethane, dimethylformamide, and dimethyl sulfoxide. The reduction processes were monitored by in situ electrochemical UV-vis spectroscopy in CH(3)CN over a range of water concentrations (0.05-10 M) to spectroscopically identify the hydrogen-bonded species. PMID- 19132834 TI - Contribution of nanoscale curvature to number density of immobilized DNA on gold nanoparticles. AB - We report the curvature size dependence of the density of attached single stranded DNA (ssDNA) on the surface of gold nanoparticles. The densities of immobilized ssDNA on 10, 20, 30, and 50 nm gold nanoparticles were examined, and we found that the maximum density of the immobilized ssDNA on 10 nm particles was 13 times larger than that on 50 nm particles, which was still 10 times larger than that on flat gold surfaces. This result indicates the importance of curvature in the nanometer-scale attachment of ssDNAs to nanoparticles. PMID- 19132835 TI - Helical nanocomposites from chiral block copolymer templates. AB - Three-dimensional hexagonally packed PLLA nanohelices in the PS matrix were formed in the self-assembly of PS-PLLA chiral block copolymer. After hydrolysis of the PLLA blocks, PS with hexagonally packed helical nanochannels can be fabricated and treated as the template for the following sol-gel process. Subsequently, silica precursor mixture was introduced into the PS template by a pore-filling process. Well-defined helical nanocomposites with SiO(2) inorganic nanohelices orderly dispersed in polymeric matrix can be successfully obtained after the sol-gel process. As a result, with the combination of the self-assembly of degradable block copolymers and sol-gel chemistry, we suggest a novel method for the preparation of the helical nanocomposites with ordered texture. PMID- 19132836 TI - Iridium-catalyzed asymmetric hydrogenation of cyclic enamines. AB - The first highly enantioselective iridium-catalyzed hydrogenation of cyclic enamines has been developed. This new reaction provided an efficient method for the synthesis of optically active cyclic tertiary amines including natural product crispine A. PMID- 19132837 TI - 4D 1H-13C NMR spectroscopy for assignments of alanine methyls in large and complex protein structures. AB - Alanine (13)CH(3) methyl groups can serve as a useful addition to the Ile, Leu, Val (ILV) selective isotope labeling methodology adopted for NMR studies of high molecular-weight protein systems. A four-dimensional (4D) methyl-detected "out and-back" NMR experiment has been developed that allowed us to obtain practically complete (1)H-(13)C assignments of more than 70 alanine methyl sites in a 723 residue enzyme Malate Synthase G. It can be anticipated that the developed NMR methodology will promote the use of alanine methyls as important probes of molecular structure and dynamics in large proteins. PMID- 19132838 TI - Adsorbate-induced defect formation and annihilation on graphene and single-walled carbon nanotubes. AB - We used density functional theory calculations to probe the chemical reactivity of graphene and single-wall carbon nanotubes (CNTs) toward the small molecules O(2), H(2), N(2), C(2)H(2), CO, and CO(2). We found that there is a threshold CNT size below which C(2)H(2) and CO, typical feedstock precursors for CNT growth, become trapped in decorated hillock-like defects on the side walls of CNTs. We also found that O(2), H(2), and CO(2) can etch isolated C adatoms and C adatom pairs. These processes play a role not only in the growth of CNTs, but also in the postgrowth evolution of defects on CNTs through exposure to typical ambient gases. PMID- 19132839 TI - Spectroscopic and calorimetric studies on the binding of alkaloids berberine, palmatine and coralyne to double stranded RNA polynucleotides. AB - The interaction of two natural protoberberine plant alkaloids berberine and palmatine and a synthetic derivative coralyne to three double stranded ribonucleic acids, poly(A). poly(U), poly(I).poly(C) and poly(C).poly(G) was studied using various biophysical techniques. Absorbance and fluorescence studies showed that the alkaloids bound cooperatively to these RNAs with the binding affinities of the order 10(4) M(-1). Circular dichroic results suggested that the conformation of poly(A). poly(U) was perturbed by all the three alkaloids, that of poly(I).poly(C) by coralyne only and that of poly(C).poly(G) by none. Fluorescence quenching studies gave evidence for partial intercalation of berberine and palmatine and complete intercalation of coralyne to these RNA duplexes. Isothermal titration calorimetric studies revealed that the binding was characterized by negative enthalpy and positive entropy changes and the affinity constants derived were in agreement with the overall binding affinity from spectral data. The binding of all the three alkaloids considerably stabilized the melting of poly(A). poly(U) and poly(I).poly(C) and the binding data evaluated from the melting data were in agreement with that obtained from other techniques. The overall binding affinity of the alkaloids to these double stranded RNAs varied in the order, berberine = palmatine < coralyne. The temperature dependence of the enthalpy changes afforded large negative values of heat capacity changes for the binding of palmatine and coralyne to poly(A).poly(U) and of coralyne to poly(I).poly(C), suggesting substantial hydrophobic contribution in the binding process. Further, enthalpy-entropy compensation was also seen in almost all the systems that showed binding. These results further advance our understanding on the binding of small molecules that are specific binders to double stranded RNA sequences. PMID- 19132840 TI - Low-temperature studies of electron transfer to the M side of YFH reaction centers from Rhodobacter capsulatus. AB - There have been extensive experimental and theoretical studies of the temperature dependence of the rates of electron transfer between the cofactors associated primarily with the L polypeptide (or A branch) in the bacterial photosynthetic reaction center (RC). The focus of this paper is to gain further insight into the temperature dependence of rate of initial electron transfer to the parallel cofactor chain associated mainly with the M polypeptide (or B branch), which is inactive in the native RC. To this end, picosecond transient absorption measurements have been carried out on RCs of the YFH mutant of the photosynthetic bacterium Rhodobacter capsulatus at 77 K. In wild-type RCs, symmetry-related residues Phe M208 and Tyr L181 flank the primary electron donor (P) and are key to initial charge separation. In the YFH mutant these residues are swapped, i.e., are Tyr M208 and Phe L181. The third mutation in YFH changes Leu M212 to His and results in replacement of the L-side bacteriopheophytin (H(L)) with a bacteriochlorophyll denoted beta. Studies were carried out at 77 K for RCs in detergent-buffer/glycerol glasses utilizing either the detergent N-lauryl-N,N dimethylamine N-oxide (LDAO) or Deriphat 160-C. In both media, excitation of P to its lowest singlet excited state (P*) elicits complex kinetic behavior that, in the simplest phenomenological description, involves two P* cofactor-protein populations: one that is capable of charge separation (active) and one that is not (inactive). The amplitudes of the components of the P* stimulated-emission kinetic profiles, and the amplitudes and time course of accompanying P-bleaching recovery, reveal that the two P* populations, active/inactive, are in 60/40 (LDAO) or 40/60 (Deriphat) proportion. In the nonphotoactive fraction, P* decays solely via return to the ground state with its inherent lifetime (i.e., lifetime in the absence of electron transfer) of 170 ps (LDAO) or 350 ps (Deriphat). In the photoactive fraction, P* has a lifetime of 4.5 ps (LDAO) or 13 ps (Deriphat) and decays by parallel electron transfer to H(M) (30%) and beta (70%) on the M and L branches, respectively. The rate constant for P* --> P(+)H(M)(-) electron transfer is (15 ps)(-1) (LDAO) or (43 ps)(-1) (Deriphat) at 77 K. These rate constants are about 2-fold greater than those determined at 295 K in the corresponding detergent/buffer solutions. These results combined with related findings from prior work show that the primary charge-separation events on both sides of the RC are basically activationless processes. In particular, in the functionally active P* population of YFH RCs, there is little or no apparent energy barrier (e.g., involving motions of the cofactors or protein or both) for electron transfer from P* to the either side of the RC. This conclusion holds irrespective of the precise description of the complex kinetic behavior that is observed. In addition to the observations on the temperature-dependent photochemistry, the 77 K transient absorption spectra in the near-infrared resolve a bacteriopheophytin anion band at 955 nm for wild-type RCs and, for the beta-containing mutants L(M212)H and YFH, a bacteriochlorophyll anion band at 1015 nm. PMID- 19132841 TI - Kinetic analysis of beta-phosphoglucomutase and its inhibition by magnesium fluoride. AB - The isomerization of beta-glucose-1-phosphate (betaG1P) to beta-glucose-6 phosphate (G6P) catalyzed by beta-phosphoglucomutase (betaPGM) has been examined using steady- and presteady-state kinetic analysis. In the presence of low concentrations of beta-glucose-1,6-bisphosphate (betaG16BP), the reaction proceeds through a Ping Pong Bi Bi mechanism with substrate inhibition (kcat = 65 s(-1), K(betaG1P) = 15 microM, K(betaG16BP) = 0.7 microM, Ki = 122 microM). If alphaG16BP is used as a cofactor, more complex kinetic behavior is observed, but the nonlinear progress curves can be fit to reveal further catalytic parameters (kcat = 74 s(-1), K(betaG1P) = 15 microM, K(betaG16BP) = 0.8 microM, Ki = 122 microM, K(alphaG16BP) = 91 microM for productive binding, K(alphaG16BP) = 21 microM for unproductive binding). These data reveal that variations in the substrate structure affect transition-state affinity (approximately 140,000-fold in terms of rate acceleration) substantially more than ground-state binding (110 fold in terms of binding affinity). When fluoride and magnesium ions are present, time-dependent inhibition of the betaPGM is observed. The concentration dependence of the parameters obtained from fitting these progress curves shows that a betaG1P x MgF3(-) x betaPGM inhibitory complex is formed under the reaction conditions. The overall stability constant for this complex is approximately 2 x 10(-16) M(5) and suggests an affinity of the MgF3(-) moiety to this transition-state analogue (TSA) of < or = 70 nM. The detailed kinetic analysis shows how a special type of TSA that does not exist in solution is assembled in the active site of an enzyme. Further experiments show that under the conditions of previous structural studies, phosphorylated glucose only persists when bound to the enzyme as the TSA. The preference for TSA formation when fluoride is present, and the hydrolysis of substrates when it is not, rules out the formation of a stable pentavalent phosphorane intermediate in the active site of betaPGM. PMID- 19132842 TI - Interaction of D-amino acid oxidase with carbon nanotubes: implications in the design of biosensors. AB - We have investigated the interaction of d-amino acid oxidase (DAAO) with single walled carbon nanotubes (CNT) by spectroscopic ellipsometry. Dynamic adsorption experiments were performed at different experimental conditions. In addition, the activity of the enzyme adsorbed at different conditions was studied. Our results indicate that DAAO can be adsorbed to CNT at different pH values and concentrations by a combination of hydrophobic and electrostatic interactions. Considering that the highest enzymatic activity was obtained by adsorbing the protein at pH 5.7 and 0.1 mg x mL(-1), our results indicate that DAAO can adopt multiple orientations on the surface, which are ultimately responsible for significant differences in catalytic activity. PMID- 19132844 TI - Magnetophotoselection in the spin-polarized triplet state radical-ion pair formed in the photo-induced solvent-mediated electron transfer reaction from N,N diethylaniline to xanthone in viscous solution. AB - The triplet state radical-ion pair (RIP) formed in the photolysis of xanthone (Xn) and N,N-diethylaniline (DEA) in a highly viscous mixtures of 2-propanol and cyclohexanol was studied by time-resolved ESR. As the viscosity of the mixed solution increases, the spectrum reveals a magnetic dipole-dipole interaction in the triplet state of the RIP. Immediately after laser photolysis, the spin polarized RIP spectrum exhibits magnetophotoselection (MPS). This suggests that the electron transfer (ET) reaction is faster than the longitudinal relaxation of the excited triplet state of Xn ((3)Xn*) or much faster than the tumbling motion of (3)Xn*. The former mechanism is likely under the conditions employed. Indeed, a huge RIP-cored aggregation is quickly formed with solvent molecules which obstruct the free revolution of the RIP. The MPS spectra also indicate that only the molecules closely located react by the solvent-mediated ET. PMID- 19132843 TI - Role of histidine-86 in the catalytic mechanism of ferredoxin:thioredoxin reductase. AB - Ferredoxin:thioredoxin reductase catalyzes the reduction of thioredoxins in plant chloroplasts using the [Fe2S2] ferredoxin as a one-electron donor and as such plays a central role in light regulation of oxygenic photosynthesis. The active site comprises a [Fe4S4] cluster next to a redox-active disulfide that is cleaved in sequential one-electron steps and the combination of spectroscopic and crystallographic studies have revealed a catalytic mechanism involving novel site specific cluster chemistry in the oxidized, one-electron- and two-electron reduced redox states. Histidine-86 has emerged as a potential proton donor/acceptor in the catalytic mechanism based on redox-related changes in the positioning of the imidazole ring during redox cycling and greatly decreased activity for the H86Y variant. Here we report on spectroscopic and redox characterization of the [Fe4S4] center in Synechocystis sp. PCC 6803 H86Y ferredoxin:thoredoxin reductase in the accessible redox states of both the as purified and N-ethylmaleimide-modified forms, using the combination of UV-visible absorption and variable-temperature magnetic circular dichroism, EPR, resonance Raman and Mossbauer spectroscopies. The results demonstrate that His86 is required for formation of the partially valence-localized [Fe4S4]2+ cluster that is the hallmark of two-electron-reduced intermediate. Taken together with the available structural data, the spectroscopic results indicate a functional role for His86 in protonation/deprotonation of the cluster-interacting thiol and anchoring the cluster interacting thiol in close proximity to the cluster in the two-electron-reduced intermediate. PMID- 19132845 TI - The pyrolytic reaction of ketonic hydrazones from S-methyl dithiocarbazate: a combined online GC-MS pyrolysis and DFT study. AB - The gas-phase pyrolysis of ketonic hydrazones from S-methyl dithiocarbazate R(1)R(2)C=N-NHC(=S)SCH(3) (R(1), R(2) = alkyl or aryl) was investigated by online GC-MS pyrolysis and theoretical calculation. Both of these pyrolytic products, ascribed to methanethiol and the corresponding N-isothiocyanate imines, were detected in the total ion chromatography (TIC) of GC-MS. Calculation results exhibit two stable configurational structures for reactants (Re), which can interconvert with relatively low barriers (<78 kJ/mol). DFT calculations showed that the two unimolecular pyrolytic processes, a direct 1,2-elimination of CH(3)SH for syn-Re and a two-step reaction pathway for trans-Re involving tautomer interconversion followed by decomposition of CH(3)SH, are competitive in the reaction. Both syn-Re and trans-Re exhibit lower critical energies in the propagation step of the radical pyrolysis than that in the unimolecular pyrolysis process (187.76 kJ/mol via 131.91 kJ/mol for syn-Re, and 159.15 kJ/mol via 98.92 kJ/mol for trans-Re). However, much more energy is needed to excite the compound to produce the methylthio radical, with 262.03 and 253.60 kJ/mol for syn-Re and trans-Re, respectively. Therefore, the unimolecular pyrolysis rather than the radical one occurs in the condition of this study. PMID- 19132846 TI - Clar theory for molecular benzenoids. AB - Eric Clar's ideas concerning "aromatic sextets" are extended to a quantitative format in terms of a polynomial called the "Clar 2-nomial", along with related derivative quantities. The quantification is successfully tested to make correlations with a selection of numerical data, including resonance energies, bond lengths, and NICS ring-aromaticity values. PMID- 19132847 TI - Effects of heteroatoms on aromatic pi-pi interactions: benzene-pyridine and pyridine dimer. AB - Heteroatoms are found in many noncovalent complexes which are of biological importance. The effect of heteroatoms on pi-pi interactions is assessed via highly accurate quantum chemical computations for the two simplest cases of interactions between aromatic molecules containing heteroatoms, namely, benzene pyridine and pyridine dimer. Benchmark quality estimated coupled-cluster through perturbative triples [CCSD(T)] binding energies are computed near the complete basis set limit. Comparisons to the benzene dimer are made to determine the contributions from heteroatoms. The presence of a heteroatom reduces the spatial extent of the pi-electron cloud and polarizability of pyridine as compared to benzene. As a result, the magnitude of the dispersion, exchange, and induction interactions in benzene-pyridine and pyridine dimer is generally reduced as compared to those for the benzene dimer. Benzene-pyridine and pyridine dimer bind more strongly than the benzene dimer in several configurations, and in contrast to the benzene dimer, parallel-displaced configurations can be significantly preferred over T-shaped configurations. Hydrogens para to a heteroatom are more effective "pi-hydrogen bond" donors, but aromatic rings with heteroatoms are worse "pi-hydrogen bond" acceptors. PMID- 19132848 TI - Onset of double helical structure in small-sized homoleptic gold thiolate clusters. AB - Low-lying isomers of homoleptic gold thiolate clusters (AuSR)(N) (N = 6-12) are studied by using a global-minimum search method. Besides previously known zigzag crown structures for (AuSR)(N) (N = 6-12) and catenane structures for (AuSR)(N) (N = 10, 12), onset of a new structural family of double helical structures in small-sized gold thiolate clusters and the separate-ring structures in larger clusters is revealed. At N = 6-9, the double helical structures are predicted to be the global minima. The onset of double helical structures suggests a new folding way for flexible gold thiolate clusters. The simulated UV-vis spectra and XRD patterns for all low-lying clusters are ready to compare with future experiments. PMID- 19132849 TI - Evaluation of urinary biomarkers for coronary artery disease, diabetes, and diabetic kidney disease. AB - BACKGROUND: In this study we sought to validate urinary biomarkers for diabetes and two common complications, coronary artery disease (CAD) and diabetic nephropathy (DN). METHODS: A CAD score calculated by summing the product of a classification coefficient and signal amplitude of 15 urinary polypeptides was previously developed. Five sequences of biomarkers in the panel were identified as fragments of collagen alpha-1(I) and alpha-1(III). Prospectively collected urine samples available for analysis from 19 out of 20 individuals with CAD (15 with type 1 diabetes [T1D] and four without diabetes) and age-, sex-, and diabetes-matched controls enrolled in the Coronary Artery Calcification in Type 1 Diabetes study were analyzed for the CAD score using capillary electrophoresis and electrospray ionization mass spectrometry. Two panels of biomarkers that were previously defined to distinguish diabetes status were analyzed to determine their relationship to T1D. Three biomarker panels developed to distinguish DN (DNS) and two biomarker panels developed to distinguish renal disease (RDS) were examined to determine their relationship with renal function. RESULTS: The CAD score was associated with CAD (odds ratio with 95% confidence interval, 2.2 [1.3 5.2]; P = 0.0016) and remained significant when adjusted individually for age, albumin excretion rate (AER), blood pressure, waist circumference, intraabdominal fat, glycosylated hemoglobin, and lipids. DNS and RDS were significantly correlated with AER, cystatin C, and serum creatinine. The biomarker panels for diabetes were both significantly associated with T1D status (P < 0.05 for both). CONCLUSIONS: We validated a urinary proteome pattern associated with CAD and urinary proteome patterns associated with T1D and DN. PMID- 19132850 TI - Evaluating the accuracy, reliability, and clinical applicability of continuous glucose monitoring (CGM): Is CGM ready for real time? AB - BACKGROUND: This study was designed to assess the accuracy, reliability, and contribution to clinical decision-making of two commercially available continuous glucose monitoring (CGM) devices using a novel analytical approach. STUDY DESIGN: Eleven individuals with type 1 diabetes and five with type 2 diabetes wore a Guardian RT (GRT) (Medtronic Minimed, Northridge, CA) or DexCom STS Continuous Monitoring System (DEX) (San Diego, CA) device for 200 h followed by an 8-h laboratory study. A subset of these subjects wore both devices simultaneously. RESULTS: Subjects produced 1,902 +/- 269 readings during the ambulatory phase. During the laboratory study we found: lag time of 21 +/- 5 min for GRT and 7 +/- 7 min for DEX (P < 0.005); mean absolute relative difference of 19.9% and 16.7%, respectively, for GRT and DEX; and glucose exposure (the ratio of study device/laboratory reference device [YSI Instruments, Inc., Yellow Springs, OH] area under the curve) of 95 +/- 6% for GRT and 101 +/- 13% for DEX. Reliability measured during laboratory study showed 82% for DEX and 99% for GRT. Clarke Error Grid analysis (YSI reference) showed for GRT 59% of values in zone A, 34% in zone B, and 7% in zone D and for DEX 70% in zone A, 28% in zone B, 1% in zone C, and 1% in zone D. Bland-Altman plots (YSI standard) yielded for DEX 3 mg/dL (95% confidence interval, -78 to 84 mg/dL) and for GRT -21 mg/dL (95% confidence interval, -124 to 82 mg/dL). Six of eight subjects completed both home and laboratory simultaneous use of DEX and GRT. Lag times were inconsistent between devices, ranging from 0 to 32 min; area under the curve revealed a tendency for DEX to report higher total glucose exposure than GRT for the same patient. CONCLUSIONS: CGM detects abnormalities in glycemic control in a manner heretofore impossible to obtain. However, our studies revealed sufficient incongruence between simultaneous laboratory blood glucose levels and interstitial fluid glucose (after calibrations) to question the fundamental assumption that interstitial fluid glucose and blood glucose could be made identical by resorting to algorithms based on concurrent blood glucose levels alone. PMID- 19132851 TI - Interactive diary for diabetes: A useful and easy-to-use new telemedicine system to support the decision-making process in type 1 diabetes. AB - BACKGROUND: Widespread use of carbohydrate (CHO) counting is limited by its complex educational needs, but a telemedicine system could simplify the patients' training. METHODS: The Diabetes Interactive Diary (DID) was set up on patients' mobile phones and allowed to record the blood glucose values and quantify the total CHO intake during a meal, by choosing the specific food and the amount ingested from a list of pictures. It also suggested the most appropriate insulin bolus in relation to the patient's CHO/insulin ratio. Data were sent to the physician by Short Message Service (also known as text message). Two pilot studies were carried out to investigate the feasibility and acceptability of the system and its effectiveness in improving metabolic control. RESULTS: In the first study, 50 patients were involved in a survey with questionnaires administered before and 12 weeks after the start of the DID. The system was considered by almost all the patients as easy to use and very helpful. CHO counting and insulin bolus calculation were ranked as the most useful functions. In the second study, 41 consecutive patients using DID under routine clinical practice conditions were evaluated after a median of 9 months of follow-up. DID was associated with a nonstatistically significant reduction in fasting blood glucose (FBG), postprandial glucose (PPG), and hemoglobin A1c levels. FBG and PPG coefficient of variation (CV) values were significantly reduced: FBG-CV decreased by 6.7% (95% confidence interval -11.9, -1.6; P = 0.02), while PPG-CV decreased by 11.5% (95% confidence interval -19.3, -3.7; P = 0.01). No patients reported serious hypoglycemic episodes requiring medical intervention. CONCLUSIONS: DID can represent a useful, safe, and easy-to-use tool to help the patient with type 1 diabetes promote dietary freedom. Adjustment of insulin doses according to CHO intake allowed the reduction of glucose variability, increasingly recognized as an important, independent risk factor for cardiovascular events. PMID- 19132852 TI - The minimal model of glucose disposal in the analysis of glucose effectiveness: importance of early insulin data. AB - BACKGROUND: Glucose effectiveness (S(g)) is an important component in glucose tolerance. Values of S(g) using "open loop" glucose kinetic computer programs are usually higher compared to closed loop method (CLM) programs that incorporate insulin secretion modeling. We aimed to test whether these differences are caused by (1) inclusion of insulin secretion modeling or (2) the method of representing plasma insulin values in the first few minutes of the frequently sampled intravenous glucose tolerance test (FSIGT). METHODS: FSIGTs without insulin supplementation were performed in six healthy volunteers, and the Bergman minimal model was fitted to the data using the simulation and modeling program SAAM. RESULTS: The CLM, which represents the insulin data in the first few minutes by a best-fit curve extrapolated to the y-axis, yielded a significantly lower S(g) than the approach similar to the computer program MINMOD, where the first few minutes of insulin data are represented by a line joining the basal to the peak values (1.55 +/- 0.28 vs. 1.97 +/- 0.27 [SE] x 10(-2)/min, P < 0.05). This second analysis was then repeated while forcing the program to represent the insulin data after the insulin peak in the same way as in the CLM, obtaining an almost identical result for S(g) (1.99 +/- 0.29). Insulin sensitivity was not significantly affected. CONCLUSIONS: The higher S(g) estimates are caused by the method of representing the first few minutes of insulin data rather than by the incorporation of insulin secretion modeling. It is, therefore, important to know how the early insulin data are represented when comparing results from different computer modeling programs. PMID- 19132853 TI - Pre- and postoperative accuracy and safety of a real-time continuous glucose monitoring system in cardiac surgical patients: a randomized pilot study. AB - BACKGROUND: Our objective was to evaluate the accuracy and safety of a real-time (RT) continuous glucose monitoring system (CGMS) in patients before and after cardiothoracic surgery and to investigate whether activation of the alarm function of the RT-CGMS had an effect on glucose control. METHODS: Patients scheduled for elective cardiothoracic procedures, without a history of insulin requiring diabetes, were perioperatively monitored with RT-CGMS for 72 h and were randomized into two groups: with or without the alarm function (set at 4 and 10 mmol/L) of the device activated. Sensor values were compared with capillary, arterial, and venous blood glucose values. Percentages of time spent in various glucose ranges were compared between groups. RESULTS: There were no adverse effects of the RT-CGMS. Of the 1,001 sensor value comparisons with capillary or arterial measurements, 96.6% fell within Clarke Error Grid zones A and B, with relative absolute differences ranging from 15% (preoperative period) to 12% (intensive care unit period) to 14% (postoperative period on the ward). Seventeen (7.9%) arterial and 16 (2.0%) capillary comparisons fell within zone D or E. Whether or not the alarm function, as used in this pilot study, was activated did not affect time spent in different glucose ranges. CONCLUSIONS: Although the RT CGMS is safe and accurate according to accepted standards, there are still small aberrations, which in our opinion preclude unlimited use in its present form in a clinical setting. The effect of the alarm function at different glucose levels remains to be investigated. PMID- 19132854 TI - The role of nitric oxide in skin blood flow increases due to vibration in healthy adults and adults with type 2 diabetes. AB - BACKGROUND: We recently demonstrated concomitant increases in skin blood flow and nitric oxide (NO) production in young healthy adults in response to externally applied vibration of the forearm. Research has shown that adults with type 2 diabetes exhibit depressed NO production and vascular responses to NO. We hypothesized that subjects with type 2 diabetes would display lower than normal increases in skin blood flow to externally applied vibration. RESEARCH DESIGN AND METHODS: Therefore, the purpose of this study was to compare 20 male and female, age- and body mass index-matched normal adults and adults with type 2 diabetes in terms of the effects of external vibration of the forearm on skin blood flow and the rate of NO production. Skin blood flow and NO production were measured before vibration, immediately after 5 min of vibration, and 5 min after vibration ceased. RESULTS: Although externally applied vibration significantly increased skin blood flow for both groups (P = 0.0001), those with diabetes had significantly lower (223%; P = 0.003) skin blood flows compared to the healthy older adults (461%). The rate of NO production, expressed as microM NO . flux, also increased significantly in both groups after vibration (healthy group, 374%; diabetes group, 236%) and remained significantly elevated (healthy group, 258%; diabetes group, 177%) for at least 5 min; however, the difference between groups was not significant (P = 0.12). CONCLUSIONS: These findings suggest that subjects with diabetes exhibit a lower skin blood flow and lower NO response to externally applied vibration than matched normal subjects. PMID- 19132855 TI - Regular visits to a diabetes clinic were associated with lower magnitude of visual disability and diabetic retinopathy-a hospital-based historical cohort study in yemen. AB - BACKGROUND: We associated regularity in visits to a diabetes clinic with the presence of diabetic retinopathy (DR) and visual disabilities. METHODS: This historical cohort study was conducted in 2004. The physician reported details of diabetes mellitus (DM), hypertension, and other illness. The ophthalmologist examined eyes and noted visual acuity, DR, and other ocular morbidities. We calculated the relative risk (RR) of different complications of diabetes. RESULTS: Our cohort consisted of 228 patients (114 in each group, one that attended diabetes clinics regularly [group A] and one that had irregular attendance [group B]). DR was found in 47 (41.2%) and 68 (61.4%) patients, respectively. The risk of DR was significantly higher in group B (RR = 1.51, 95% confidence interval [CI] 1.23 to 2.18). The severity of DR was also positively associated with irregularity in clinic visits (x(2) = 33.56, degrees of freedom = 5, P = 0.000003). The risk of bilateral blindness (RR = 4.0, 95% CI 1.38 to 11.6) and low vision disability (RR = 2.53, 95% CI 1.84 to 3.47) were higher in group B. The duration of diabetes and the regularity in clinic visits were the predictors of DR. CONCLUSIONS: The presence of DR and visual disabilities among patients with diabetes is associated with irregular attendance at diabetes clinics. The regularity of medical visits seems to be a proxy indicator of better primary prevention of eye complications of DM. PMID- 19132856 TI - The effect of insulin lispro on glycemic control in a large patient cohort. AB - BACKGROUND: The use of rapid-acting insulin analogs and regular insulin differs considerably in countries throughout the world. We therefore studied how glycemic control has been affected by using insulin lispro in clinical practice over 5 years in 14 hospitals in Sweden. METHODS: We used a time period when most patients had not changed the basal insulin, but only the mealtime insulin. Accordingly the most recent years were not suitable since many patients had changed basal insulin from NPH to glargine or determir. We therefore analyzed the metabolic consequences on glycosylated hemoglobin (HbA1c) when changing from regular insulin to insulin lispro from 1997 and during the following 5 years. We studied 1,069 patients with diabetes taking NPH insulin as basal insulin and at least three daily injections of regular insulin, of whom 423 changed their mealtime insulin to insulin lispro and 646 controls who continued with regular insulin. RESULTS: Patients changing to insulin lispro on average decreased by 0.19% units more in HbA1c than those remaining on regular insulin. The effect was most pronounced in patients with high HbA1c even after controlling for regression to the mean. A beneficial effect of insulin lispro was also indicated since patients had the same level of HbA1c during a long period of time with regular insulin but then dropped when changing to insulin lispro. CONCLUSIONS: This study indicates that insulin lispro has had a beneficial and persisting effect on glycemic control when used in patients with diabetes on multiple daily injections of insulin in clinical practice. PMID- 19132857 TI - Patient-reported outcomes for an integrated real-time continuous glucose monitoring/insulin pump system. AB - BACKGROUND: A 16-week, two-site study evaluated outcomes for a new device (the Paradigm 722 System, Medtronic MiniMed, Northridge, CA) that combines a "smart" continuous subcutaneous insulin infusion (CSII) pump with real-time (RT) continuous glucose monitoring (CGM) and CareLinktrade mark data management software (DMS). METHODS: CSII-naive adults with type 1 diabetes in suboptimal control (mean glycosylated hemoglobin [A1C] = 8.6%) were randomized to the control arm, consisting of multiple daily injections (MDI) and self-monitoring of blood glucose (SMBG), or the study arm (CSII with RT-CGM as an adjunct to SMBG). Participants (n = 28) completed the validated Insulin Delivery System Rating Questionnaire (IDSRQ) and the parallel Blood Glucose (BG) Monitoring System Rating Questionnaire (BGMSRQ) at study start and end. Participants in the study arm (n = 14) also completed newly developed User Acceptance Questionnaires (UAQs) for CSII, RT-CGM, and DMS at study end. RESULTS: A1C reduction from study start to end was significant (P < 0.05) in both arms (-1.7% for study arm;-1.0% for control arm); there was no significant change in weight in either arm. The IDSRQ showed significantly (P < 0.05) greater benefit for the study arm in convenience, acceptability of BG monitoring requirements, BG control efficacy, diabetes worries, and interpersonal hassles, as well as higher overall satisfaction/preference. The BGMSRQ showed significantly (P < 0.05) greater benefit for the study arm in the BG monitoring system's ability to help manage glycemic control and less interest in changing to another BG monitoring system. The Study Arm UAQs showed positive ratings of system features. CONCLUSIONS: Several patient-reported outcomes were significantly more positive in the study arm than the control arm; none was significantly more positive in the control arm. The features of the integrated RT-CGM/CSII system were frequently used and highly rated by participants, with high user satisfaction. PMID- 19132858 TI - Biofunctional surfaces based on dendronized cellulose. AB - Biofunctionalized surfaces based on dendronized cellulose were prepared either by embedding 6-deoxy-6-(1,2,3-triazolo)-4-polyamidoamine (PAMAM) cellulose (degree of substitution, DS 0.25), obtained by homogeneous conversion of 6-deoxy-6-azido cellulose with propargyl-PAMAM dendron via the copper-catalyzed Huisgen reaction, in a cellulose acetate (DS 2.50) matrix or by the heterogeneous functionalization of deoxy-azido cellulose film with the dendron. The amount of amino groups provided by the solid supports was determined and the covalent attachment of enzyme was proven with glucose oxidase as model enzyme after activation with glutardialdehyde. The quality of glucose oxidase immobilization was defined by determining of the specific enzyme activity, coupling efficiency, storage stability, and reproducibility. Although the heterogeneous functionalization of the deoxy-azido film yields a product that binds more enzyme compared to the blend of dendronized cellulose derivative with cellulose acetate, the coupling efficiency is comparatively small. Nevertheless, the different approaches for the preparation of biofunctionalized surfaces based on dendronized cellulose provide an excellent reproducibility and good storage stability. PMID- 19132859 TI - Aroma permeability of hydroxypropyl maize starch films. AB - In this study, the role of water and the impact of the glass to rubber transition on aroma mass transport through hydroxypropyl maize starch films were followed. The permeability of four aroma compounds (ethanol, pyrazine, menthone, and decanone) was monitored by atmospheric pressure chemical ionization mass spectroscopy. The increase in water content within the film promoted greater water diffusion and film flexibility and resulted in enhanced transport of the four aroma compounds through the film. At low water contents in the glassy state, the permeability of ethanol was much higher than those of the other three compounds, which was attributed to its low molecular volume and greater solubility in starch. The structural changes induced by the glass transition resulted in the hydroxypropyl maize starch films displaying poor barrier behavior in the rubbery state for all the aromas studied. PMID- 19132860 TI - General quantum-mechanical study on the hydrolysis equilibria for a tetravalent aquaion: the extreme case of the Po(IV) in water. AB - A systematic study of the different hydrolyzed species derived from the hydrated Po(IV) in water, [Po(H(2)O)(n)(OH)(m)]((4-m)) for 1 m 4, and 4 m + n 9, has been carried out by means of quantum mechanical computations. The effects of outer solvation shells have been included using a polarizable continuum dielectric model. For a fixed number of hydroxyl groups, the preferred hydration number for the Po(IV) can be determined in terms of Gibbs energy. It is shown that the hydration number (n) systematically decreases with the increase in the number of hydroxyl groups (m) in such a way the total coordination number (n + m) becomes smaller, being 9 in the aquocomplex and 4 in the neutral hydroxo-complex. Free energies for the hydrolysis processes involving Po(IV) complexes and a different number of hydroxyl groups have been computed, revealing the strong tendency toward hydrolysis exhibited by these complexes. The predominant species of Po(IV) in aqueous solutions are ruled by a dynamical equilibrium involving aggregates containing in the first coordination shell OH(-) groups and water molecules. Although there is not experimental information to check the theoretical predictions, theoretical computations in solution seem to suggest that the most likely clusters are [Po(H(2)O)(5)(OH)(2)](2+) and [Po(H(2)O)(4)(OH)(2)](2+). The geometry of the different clusters is ruled by the trend of hydroxyl groups to be mutually orthogonal and to promote a strong perturbation of the water molecule in trans-position by lengthening the Po-H(2)O distances and tilting the corresponding bond angle. A general thermodynamic cycle is defined to compute the Gibbs free energy associated to the formation of the different hydrolyzed forms in solution. From it, the estimates of pK(a) values associated to the different protolytic equilibria are provided and discussed. Comparison of the relative values of pK(a) along a hydrolysis series with the experimental values for other tetravalent cations supports its consistency. PMID- 19132861 TI - Origin of the activity drop with the E50D variant of catalytic antibody 34E4 for Kemp elimination. AB - In enzymes, multiple structural effects cooperatively lead to the high catalytic activity, while individually these effects can be small. The design of artificial enzymes requires the understanding and ability to manipulate such subtle effects. The 34E4 catalytic antibody, catalyzing Kemp elimination of 5-nitrobenzisoxazole, and its Glu50Asp (E50D) variant are the subject of the present investigation. This removal of only a methylene group yields an approximately 30-fold reduction in the rate for the catalyzed Kemp elimination. Here, the aim is to understand this difference in the catalytic performance. The mechanism of Kemp elimination catalyzed by 34E4 and the E50D mutant is elucidated using QM/MM Monte Carlo simulations and free energy perturbation theory. In both proteins, the reaction is shown to follow a single-step, concerted mechanism. In the mutant, the activation barrier rises by 2.4 kcal/mol, which corresponds to a 62-fold rate deceleration, which is in good agreement with the experimental data. The positions and functionality of the residues in the active site are monitored throughout the reaction. It is concluded that the looser contact with the base, shorter base-Asn58 contact, less favorable pi-stacking with Trp91 in the transition state of the reaction, and different solvation pattern all contribute to the reduction of the reaction rate in the E50D variant of 34E4. PMID- 19132863 TI - Steroids from an Australian sponge Psammoclema sp. AB - Investigation of an extract of the Australian marine sponge Psammoclema sp. for dynamin I inhibitory activity led to the isolation of four new trihydroxysterols (1-4) related to aragusterol G. These compounds were largely identified by 1D and 2D NMR spectroscopic methods. While 1 was found to be inactive in the dynamin bioassay, bioassays did reveal that compounds 1-4 inhibited the growth of colorectal, breast, ovarian, and prostate cancer cell lines (GI(50) 5-27 microM). The additional insight that these new compounds give to previous SAR studies is discussed briefly. PMID- 19132862 TI - Effect of dehydration on the aggregation kinetics of two amyloid peptides. AB - It is well-known that water plays a crucial role in the folding, dynamics, and function of proteins. Here we provide further evidence showing that the aggregation kinetics of peptides also depend strongly on their hydration status. Using reverse micelles as a tool to modulate the accessible number of water molecules and infrared spectroscopy and transmission electron microscopy as means to monitor aggregate formation, we show that the rate of aggregation of two amyloid forming peptides increases significantly under conditions where limited hydration of the peptide molecule is expected to occur. These results not only are in accord with recent computer simulations indicating that the expulsion of interfacial water molecules is a key event in the dimerization/oligmerization of amyloid beta (Abeta) peptides but also have implications for amyloid formation in vivo where molecular crowding is expected to influence the solvation status of proteins. PMID- 19132864 TI - Emerging concepts in effector biology of plant-associated organisms. AB - Plant-associated organisms secrete proteins and other molecules to modulate plant defense circuitry and enable colonization of plant tissue. Understanding the molecular function of these secreted molecules, collectively known as effectors, became widely accepted as essential for a mechanistic understanding of the processes underlying plant colonization. This review summarizes recent findings in the field of effector biology and highlights the common concepts that have emerged from the study of cellular plant pathogen effectors. PMID- 19132865 TI - Development and use of an efficient DNA-based viral gene silencing vector for soybean. AB - Virus-induced gene silencing (VIGS) is increasingly being used as a reverse genetics tool to study functions of specific plant genes. It is especially useful for plants, such as soybean, that are recalcitrant to transformation. Previously, Bean pod mottle virus (BPMV) was shown to be an effective VIGS vector for soybean. However, the reported BPMV vector requires in vitro RNA transcription and inoculation, which is not reliable or amenable to high-throughput applications. To increase the efficiency of the BPMV vector for soybean functional genomics, a DNA-based version was developed. Reported here is the construction of a Cauliflower mosaic virus 35S promoter-driven BPMV vector that is efficient for the study of soybean gene function. The selection of a mild rather than a severe BPMV strain greatly reduced the symptom interference caused by virus infection. The DNA-based BPMV vector was used to silence soybean homologues of genes involved in plant defense, translation, and the cytoskeleton in shoots and in roots. VIGS of the Actin gene resulted in reduced numbers of Soybean mosaic virus infection foci. The results demonstrate the utility of this new vector as an efficient tool for a wide range of applications for soybean functional genomics. PMID- 19132866 TI - Transformed hairy roots of Arachis hypogea: a tool for studying root nodule symbiosis in a non-infection thread legume of the Aeschynomeneae tribe. AB - Arachis hypogea is a non-"infection thread" (IT) legume where rhizobial entry or dissemination in the nodules never involves IT. Rhizobia invade through epidermal "cracks" and directly access the cortical cells to develop the characteristic aeschynomenoid nodules. For investigating these nonclassical nodulation features in Arachis spp., we developed an efficient procedure for Agrobacterium rhizogenes R1000-mediated transformation of this plant. In this study, we optimized the induction of hairy roots and nodulation of composite Arachis hypogea plants in the presence of Bradyrhizobium sp. (Arachis) strain NC92. 35S promoter-driven green fluorescent protein and beta-glucuronidase expression indicated transformation frequency to be above 80%. The transformed roots had the characteristic rosette-type root hairs and had normal level of expression of symbiosis-related genes SymRK and CCaMK. The transgenic nodules resembled the wild-type nodules with an exception of 2 to 3%, where they structurally deviated from the wild-type nodules to form nodular roots. A 16S rRNA profile of an infected-zone metagenome indicated that identical populations of bradyrhizobia invaded both composite wild-type plants grown in natural soil. Our results demonstrate that Arachis hairy root is an attractive system for undertaking investigations of the nonclassical features associated with its nitrogen-fixing symbiotic interactions. PMID- 19132867 TI - Discovery of pathogenicity genes in the crucifer anthracnose fungus Colletotrichum higginsianum, using random insertional mutagenesis. AB - Agrobacterium tumefaciens-mediated transformation (ATMT) was used for random insertional mutagenesis to identify pathogenicity genes in the hemibiotrophic fungus Colletotrichum higginsianum. A high-throughput primary infection assay on Arabidopsis thaliana seedlings allowed the rapid screening of 8,850 transformants. Forty mutants showing reproducible pathogenicity defects on Arabidopsis and Brassica plants were obtained, and their infection phenotypes were characterized microscopically. Six mutants were impaired in appressorial melanization, fifteen had reduced penetration ability, 14 induced host papillae or hypersensitive cell death, and five were affected in the transition from biotrophy to necrotrophy. Southern blot analysis showed 58% of the transformants had single-site T-DNA integrations. Right-border flanking sequences were recovered from 12 mutants by inverse polymerase chain reaction (PCR) or thermal asymmetric interlaced PCR and were used to isolate the tagged genes from a genomic library. The putative pathogenicity genes encoded homologs of a major facilitator superfamily phosphate transporter, importin-beta2, ornithine decarboxylase, beta-1,3(4)-glucanase, ATP-binding endoribonuclease, carbamoyl phosphate synthetase, and the polyprotein precursor of N-acetylglutamate kinase and N-acetylglutamyl-phosphate reductase. Six further loci were homologous to proteins of unknown function. None of these genes were previously implicated in the pathogenicity of any Colletotrichum species. The results demonstrate that ATMT is an effective tool for gene discovery in this model pathogen. PMID- 19132868 TI - The TIR domain of TIR-NB-LRR resistance proteins is a signaling domain involved in cell death induction. AB - In plants, the TIR (toll interleukin 1 receptor) domain is found almost exclusively in nucleotide-binding (NB) leucine-rich repeat resistance proteins and their truncated homologs, and has been proposed to play a signaling role during resistance responses mediated by TIR containing R proteins. Transient expression in Nicotiana benthamiana leaves of "TIR + 80", the RPS4 truncation without the NB-ARC domain, leads to EDS1-, SGT1-, and HSP90-dependent cell death. Transgenic Arabidopsis plants expressing the RPS4 TIR+80 from either dexamethasone or estradiol-inducible promoters display inducer-dependent cell death. Cell death is also elicited by transient expression of similarly truncated constructs from two other R proteins, RPP1A and At4g19530, but is not elicited by similar constructs representing RPP2A and RPP2B proteins. Site-directed mutagenesis of the RPS4 TIR domain identified many loss-of-function mutations but also revealed several gain-of function substitutions. Lack of cell death induction by the E160A substitution suggests that amino acids outside of the TIR domain contribute to cell death signaling in addition to the TIR domain itself. This is consistent with previous observations that the TIR domain itself is insufficient to induce cell death upon transient expression. PMID- 19132869 TI - Activation of the salicylic acid signaling pathway enhances Clover yellow vein virus virulence in susceptible pea cultivars. AB - The wild-type strain (Cl-WT) of Clover yellow vein virus (ClYVV) systemically induces cell death in pea cv. Plant introduction (PI) 118501 but not in PI 226564. A single incompletely dominant gene, Cyn1, controls systemic cell death in PI 118501. Here, we show that activation of the salicylic acid (SA) signaling pathway enhances ClYVV virulence in susceptible pea cultivars. The kinetics of virus accumulation was not significantly different between PI 118501 (Cyn1) and PI 226564 (cyn1); however, the SA-responsive chitinase gene (SA-CHI) and the hypersensitive response (HR)-related gene homologous to tobacco HSR203J were induced only in PI 118501 (Cyn1). Two mutant viruses with mutations in P1/HCPro, which is an RNA-silencing suppressor, reduced the ability to induce cell death and SA-CHI expression. The application of SA and of its analog benzo (1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (BTH) partially complemented the reduced virulence of mutant viruses. These results suggest that high activation of the SA signaling pathway is required for ClYVV virulence. Interestingly, BTH could enhance Cl-WT symptoms in PI 226564 (cyn1). However, it could not enhance symptoms induced by White clover mosaic virus and Bean yellow mosaic virus. Our report suggests that the SA signaling pathway has opposing functions in compatible interactions, depending on the virus-host combination. PMID- 19132870 TI - Allelic variants of the Pseudomonas syringae type III effector HopZ1 are differentially recognized by plant resistance systems. AB - The bacterial plant pathogen Pseudomonas syringae depends on the type III secretion system and type III-secreted effectors to cause disease in plants. HopZ is a diverse family of type III effectors widely distributed in P. syringae isolates. Among the HopZ homologs, HopZ1 is ancient to P. syringae and has been shown to be under strong positive selection driven by plant resistance-imposed selective pressure. Here, we characterized the virulence and avirulence functions of the three HopZ1 alleles in soybean and Nicotiana benthamiana. In soybean, HopZ1 alleles have distinct functions: HopZ1a triggers defense response, HopZ1b promotes bacterial growth, and HopZ1c has no observable effect. In N. benthamiana, HopZ1a and HopZ1b both induce plant defense responses. However, they appear to trigger different resistance pathways, evidenced by two major differences between HopZ1a- and HopZ1b-triggered hypersensitive response (HR): i) the putative N-acylation sites had no effect on HopZ1a-triggered cell death, whereas it greatly enhanced HopZ1b-triggered cell death; and ii) the HopZ1b triggered HR, but not the HopZ1a-triggered HR, was suppressed by another HopZ homolog, HopZ3. We previously demonstrated that HopZ1a most resembled the ancestral allelic form of HopZ1; therefore, this new evidence suggested that differentiated resistance systems have evolved in plant hosts to adapt to HopZ1 diversification in P. syringae. PMID- 19132871 TI - Transcriptome profiling in hybrid poplar following interactions with Melampsora rust fungi. AB - In natural conditions, plants are subjected to a combination of biotic stresses and often have to cope with simultaneous pathogen infections. In this report, we aim to understand the global transcriptional response of hybrid poplar NM6 (Populus nigra x P. maximowiczii) to infection by two biotrophic Melampsora fungi, Melampsora larici-populina and M. medusae f. sp. deltoidae. These pathogens triggered different responses after inoculation of poplar leaves. Transcript profiling using the GeneChip Poplar Genome Array revealed a total of 416 differentially expressed transcripts whose expression level was > or = twofold relative to controls. Interestingly, approximately half of the differentially expressed genes in infected leaves showed altered expression following interaction with either of the Melampsora spp. We also infected poplar leaves simultaneously with both Melampsora spp. to investigate potential interaction between the responses to the individual pathogens during a mixed infection. For this mixed inoculation, the number of differentially expressed transcripts increased to 648 and our analysis showed that infection with both fungi also induced a common set of genes. The genes induced after Melampsora spp. infection were mainly related to primary and secondary metabolic processes, cell wall reinforcement and lignification, defense and stress-related mechanisms, and signal perception and transduction. PMID- 19132872 TI - Constitutive expression of OsGH3.1 reduces auxin content and enhances defense response and resistance to a fungal pathogen in rice. AB - GH3 genes are main components of the hormonal mechanism regulating growth and development and, hence, are deeply involved in a broad range of physiological processes. They are implicated in hormonal homeostasis through the conjugation to amino acids of the free form of essential plant growth regulators such as indoleacetic and jasmonic acids. In this work, we showed that OsGH3.1 overexpression in rice caused dwarfism and significantly reduced both free auxin content and cell elongation. Functional classification of the transcriptomic profiling revealed that most genes involved in auxin biosynthesis and auxin signaling inhibition were induced and repressed, respectively. Many genes related to cell organization and biogenesis were also significantly downregulated. The survey also showed that, although the response to abiotic stresses was not clearly stimulated, OsGH3.1 overexpression did activate a significant number of defense-related genes. In successive bioassays, it was demonstrated that the resistance of rice plants to pathogen infection, evaluated with two different Magnaporthe grisea strains, was higher in the transformants overexpressing OsGH3.1. Taken together, these results indicate that OsGH3.1 overexpression reduces auxin content, inhibits cell growth and cell wall loosening, and enhances resistance to a fungal pathogen. Our results provide evidence that auxin homeostasis can regulate the activation of the defense response in rice. PMID- 19132873 TI - Silencing of Kex2 significantly diminishes the virulence of Cryphonectria parasitica. AB - Cryphonectria parasitica is the causal agent of chestnut blight. Infection of this ascomycete with Cryphonectria hypovirus 1 (CHV1) results in reduction of virulence and sporulation of the fungus. The virus affects fungal gene expression and several of the CHV1 downregulated genes encode secreted proteins that contain consensus Kex2 processing signals. Additionally, CHV1 has been shown to colocalize in infected cells primarily with fungal trans-Golgi network vesicles containing the Kex2 protease. We report here the cloning, analysis, and possible role of the C. parasitica Kex2 gene (CpKex2). CpKex2 gene sequence analysis showed high similarity to other ascomycete kexin-like proteins. Southern blot analyses of CpKex2 showed a single copy of this gene in the fungal genome. In order to monitor the expression and evaluate the function of CpKex2, antibodies were raised against expressed protein and Kex2-silenced mutants were generated. Western blots indicate that the Kex2 protein was constitutively expressed. Growth rate of the fungus was not significantly affected in Kex2-silenced strains; however, these strains showed reduced virulence, reduced sexual and asexual sporulation, and reductions in mating and fertility. The reduced virulence was correlated with reduced Kex2 enzymatic activity and reduced relative mRNA transcript levels as measured by real time reverse-transcriptase polymerase chain reaction. These results suggest that secreted proteins processed by Kex2 are important in fungal development and virulence. PMID- 19132874 TI - Inactivation of the lipoxygenase ZmLOX3 increases susceptibility of maize to Aspergillus spp. AB - Plant and fungal lipoxygenases (LOX) catalyze the oxidation of polyunsaturated fatty acids, creating fatty-acid hydroperoxides (oxylipins). Fungal oxylipins are required for normal fungal development and secondary metabolism, and plant host derived oxylipins interfere with these processes in fungi, presumably by signal mimicry. The maize LOX gene ZmLOX3 has been implicated previously in seed Aspergillus interactions, so we tested the interactions of a mutant maize line (lox3-4, in which ZmLOX3 is disrupted) with the mycotoxigenic seed-infecting fungi Aspergillus flavus and Aspergillus nidulans. The lox3-4 mutant was more susceptible than wild-type maize to both Aspergillus species. All strains of A. flavus and A. nidulans produced more conidia and aflatoxin (or the precursor sterigmatocystin) on lox3-4 kernels than on wild-type kernels, in vitro and under field conditions. Although oxylipins did not differ detectably between A. flavus infected kernels of the lox3-4 and wild-type (WT) maize, oxylipin precursors (free fatty acids) and a downstream metabolite (jasmonic acid) accumulated to greater levels in lox3-4 than in WT kernels. The increased resistance of the lox3 4 mutant to other fungal pathogens (Fusarium, Colletotrichum, Cochliobolus, and Exserohilum spp.) is in sharp contrast to results described herein for Aspergillus spp., suggesting that outcomes of LOX-governed host-pathogen interactions are pathogen-specific. PMID- 19132875 TI - Temperature-induced microstructural changes in ionic liquid-based microemulsions. AB - In the present contribution, results concerning the effect of temperature on the nonionic surfactant Triton X-100 based 1-butyl-3-methylimidazolium tetrafluoroborate (bmimBF4)-in-cyclohexane and bmimBF4-in-toluene ionic liquid (IL) reverse microemulsions are reported. Dynamic light scattering (DLS) along with freeze-fracture transmission electron microscopy (FF-TEM) measurements revealed that the sizes of single microemulsion droplets increased with increasing temperature. However, a decreased temperature led to the appearance of droplet clusters, which have also been observed previously when the single microemulsion droplets were swollen by added bmimBF4 to a certain extent (Gao, Y. A.; Vogit, A.; Hilfert, L.; Sundmacher, K. ChemPhysChem, 2008, 9, 1603-1609). Compared to traditional aqueous microemulsions, IL microemulsions revealed relatively high temperature-independence. The droplet-shaped microstructure was always kept in a large range of temperature. The temperature-independence is ascribed to the temperature-insensitive electrostatic attraction between the solubilized bmimBF4 and Triton X-100, which was considered to be the driving force for solubilizing bmimBF4 into the cores of Triton X-100 aggregates. Two dimensional rotating frame nuclear Overhauser effect (NOE) experiments (ROESY) further confirmed the microstructural change with temperature. PMID- 19132876 TI - Simulated mutagenesis of the hypervariable loops of a llama VHH domain for the recovery of canonical conformations. AB - In this work, wildtype and mutated hypervariable regions of an anti-hCG llama VHH antibody were simulated via a molecular dynamics replica exchange method (REM). Seven mutants were simulated with the goal of identifying structural determinants that return the noncanonical H1 loop of the wildtype antibody to the type 1 canonical structure predicted by database methods formulated for conventional antibodies. Two cases with three point mutations yielded a stable type 1 H1 structure. In addition, other mutants with fewer mutations showed evidence of such conformations. Overall, the mutagenesis results suggest a marked influence of interloop interactions on the attainment of canonical conformations for this antibody. On the methodological front, a novel REM scheme was developed to quickly screen diverse mutants based on their relative propensities for attaining favorable structures. This multimutant REM (MMREM) was used to successfully identify mutations that stabilize a canonical H1 loop grafted on the llama antibody scaffold. The use of MMREM and REM for screening mutants and assessing structural stability may be useful in the rational design of antibody hypervariable loops. PMID- 19132877 TI - Precipitation of oppositely charged nanoparticles by dilution and/or temperature increase. AB - Mixtures of oppositely charged nanoparticles (NPs) exhibit anomalous solubility behavior and precipitate either upon dilution or upon temperature increase. Precipitation is reversible and can be explained by a thermodynamic model that accounts for changes in the electrostatic interactions due to the adsorption/desorption of counterions from the surface of the NPs. Specifically, decreasing the salt concentration via dilution or increasing the temperature causes dissociation of counterions from the NP surfaces, increasing the magnitude of electrostatic interactions between NPs and resulting in their precipitation. Model predictions of NP solubility are in quantitative agreement with the experimental observations. Such predictions are of practical importance for the preparation of "patchy" electrostatic coatings and ionic-like NP supracrystals. PMID- 19132878 TI - Zinc fingers as biologic redox switches? AB - Despite zinc ions being redox inert in biologic systems, zinc-finger structures act as redox-sensitive molecular switches controlling several crucial cellular processes. Oxidative or nitrosative stress, via modification of zinc finger cysteine thiols, leads to a release of Zn(2+) from these structures, causing not only a loss of zinc-finger function but also an increase of cytoplasmic or nuclear free Zn(2+) that may, in turn, stimulate and interfere with cellular signaling cascades. A signaling cascade stimulated by exposure of cells to zinc ions or to stressful stimuli that are reported to cause an intracellular release of zinc ions involves phosphoinositide 3'-kinases and the Ser/Thr protein kinase Akt, resulting in an inactivation of transcriptional regulators of the FoxO family. Possible modes of action of zinc ions to stimulate this signaling cascade and consequences of stimulation are discussed. Moreover, we present an overview on human diseases or disorders characterized by an intracellular Zn(2+) dyshomeostasis. PMID- 19132880 TI - Clinical characteristics and medication use among premenopausal women with osteoporosis and low BMD: the experience of an osteoporosis referral center. AB - AIMS: Osteoporosis is uncommon in premenopausal women, and most cases have a secondary cause. Women with osteoporosis and no known secondary cause are said to have idiopathic osteoporosis (IOP). We aimed to estimate the proportion of premenopausal women seen in our referral center with IOP as opposed to secondary osteoporosis, to describe their clinical characteristics, to compare women with a low-trauma fracture history with those with low bone mineral density (BMD) alone, and to estimate the frequency of bisphosphonate use. METHODS: We reviewed medical records from all premenopausal women evaluated for osteoporosis or low BMD in our center during 2005. We included premenopausal women diagnosed on the basis of low trauma fracture, low BMD or both (Z score < or= -2.0 or T score < or = -2.5), or both. RESULTS: Among these patients (n = 61; mean age 37 +/- 8), 57 (93%) were Caucasian, 34 (57%) had a family history of osteoporosis, and 26 (43%) had used bisphosphonates. The most common secondary causes were amenorrhea (34%, n = 21), anorexia nervosa (16%, n = 10), and glucocorticoid exposure (13%, n = 8). After exclusion of secondary causes, 39% (24 of 61) of the entire group and 48% (14 of 29) of the fracture group were thought to have IOP. Women with a known secondary cause had lower BMD Z scores at the spine and hip than those with IOP. Women with low BMD and no fractures had shorter stature and weighed less than those with fractures, but overall differences between the groups were not statistically significant. Bisphosphonates had been prescribed for 38% (11 of 29) of women with a fracture history and 47% (15 of 32) of women with low BMD and no fractures. CONCLUSIONS: Our findings suggest that IOP is common among premenopausal women with osteoporosis or low BMD evaluated at a referral center. The smaller stature of women diagnosed only on the basis of BMD criteria raises the question of whether their areal BMD measurements are spuriously low because of smaller bone size. The high proportion of premenopausal women who had been prescribed oral bisphosphonates for low BMD measurements is of concern, as such women are likely to be at low short-term risk of fracture, and a more conservative approach to therapy is preferable in this group. PMID- 19132881 TI - Does maternal role functioning improve with antidepressant treatment in women with postpartum depression? AB - AIMS: The ability to mother her infant is reduced in a woman with postpartum depression (PPD). Although antidepressant treatment effectively improves depressive symptoms, various domains of functioning, for example, work and relationships, do not universally improve with treatment. In this pilot study, we investigated whether maternal role functioning improved with antidepressant treatment in women with PPD. METHODS: The pilot study was an exploratory analysis of a larger study. A subset of women (n = 27) from a randomized clinical trial (double-blind, 8-week trial of nortriptyline compared with sertraline) completed three outcome measures of maternal role functioning: gratification in the maternal role, the Infant Care Survey (ICS), and videotapes of maternal-infant interaction. The tapes were analyzed using the Child and Caregiver Mutual Regulation Coding Scale and Noldus Behavioral Coding Software. RESULTS: The two antidepressants were equally efficacious in decreasing depressive symptoms and improving overall functioning and gratification in the maternal role. Differences between times 1 and 2 in the mother-infant interactions were related to time (increasing age of the infant) and not assignment of antidepressant or remission of depression. CONCLUSIONS: Effective treatment with two antidepressants improves gratification in the maternal role but not self-efficacy or maternal-infant interaction in women with PPD. Results of the study can help women and their healthcare providers to weigh the benefits of short-term antidepressant treatment in the postpartum period. Future studies should consider outcomes related to a longer duration of treatment. PMID- 19132882 TI - Dose-response relationship between moderate-intensity exercise duration and coronary heart disease risk factors in postmenopausal women. AB - PURPOSE: This study was designed to investigate whether, in a dose-response manner, there would be greater health benefits in a group of postmenopausal women completing 45 minute- vs. 30 minutes of moderate intensity (50% maximal oxygen uptake reserve, VO2R) exercise 5 days . wk(-1). METHODS: Apparently healthy but sedentary postmenopausal women (n = 33) were randomized to a nonexercise control group, a 30-minute exercise duration group, or a 45-minute exercise duration group. Exercise training was performed 5 days . wk(-1) for 12 weeks at 50% VO2R. Participants were instructed to not change their usual diet throughout the study. RESULTS: Twenty-six women completed the study. After 12 weeks, VO2max increased significantly (p < 0.05) in both 30-minute (0.20 +/- 0.21 L . min(-1)) and 45 minute (0.41 +/- 0.10 L . min(-1)) groups. Repeated measures ANOVA identified a significant interaction between exercise duration and VO2max values (F = 4.72, p < 0.05), indicating that VO2max responded differently to 30-minute and 45-minute exercise durations. Trend analysis showed that body mass, body composition, waist circumference, and high-density lipoprotein cholesterol (HDL-C) changed favorably (p < 0.05) across control, 30-minute, and 45-minute groups. CONCLUSIONS: Although most health organizations agree that 150 min . wk(-1) of physical activity will reduce the risk of all-cause and cardiovascular mortality, few randomized, controlled studies have examined whether completing more physical activity than the recommended amount will yield additional benefits. Findings from the present study suggest that there is a dose-response relationship between exercise duration and numerous health outcomes in postmenopausal women, including cardiorespiratory fitness, body mass, body composition, waist circumference, and HDL-C. PMID- 19132883 TI - Past traumatic events: are they a risk factor for high-risk pregnancy, delivery complications, and postpartum posttraumatic symptoms? AB - BACKGROUND: This study aimed to examine the association among past traumatic events, high-risk pregnancy, delivery complications, and postpartum posttraumatic stress (PTS) symptoms. METHODS: The final convenience sample consisted of 1071 Jewish women at midpregnancy. Data were gathered at three time points (during pregnancy and 1 month and 6 months after childbirth) through self-report questionnaires. RESULTS: There was a higher percentage of high-risk pregnancy among those who reported a history of traumatic events. Although the total score of PTS symptoms did not correspond with high-risk pregnancy, the intrusion and avoidance subscales did. Furthermore, a history of traumatic events as well as prenatal PTS symptoms, prenatal depression, and the subjective pain and distress during delivery accounted for postpartum PTS symptoms. Prenatal depression was found to account for delivery complications. CONCLUSIONS: Findings indicate that a history of trauma should be considered a risk factor for high-risk pregnancy and for postpartum PTS symptoms. PMID- 19132885 TI - Powerful reactive sorption of silver(I) and mercury(II) onto poly(o phenylenediamine) microparticles. AB - The strong adsorbability of Ag(I) and Hg(II) ions onto fine poly(o phenylenediamine) (PoPD) microparticles synthesized through a chemically oxidative polymerization of o-phenylenediamine was systematically examined and PoPD/Ag nanocomposites were facilely prepared through the reactive sorption method. The effect of the (NH4)2S2O8 oxidant/o-phenylenediamine monomer ratio on the polymerization yield, macromolecular structure, conductivity, and insolubility of the PoPD microparticles was studied. The Ag(I) adsorbability of the microparticles was significantly optimized by varying the oxidant/monomer ratio, doping state, Ag(I) concentration, sorption time, and solution pH. The Ag(I) adsorbance steadily increases with changing oxidant/monomer molar ratio from 3/1 to 1/1, reaching up to the highest Ag(I) adsorbance of 533 mg.g(-1) at the oxidant/monomer ratio of 1/1. The sorption process fits the pseudosecond order kinetics. The sorption is rapid because both the adsorbance and adsorptivity within 30 min reach up to 76% of the final values. The initial sorption rate of silver ions obtained from the pseudosecond-order equation is 12.9 mg.g(-1).min(-1). The highest adsorptivity of silver ions is up to 99.1%. The optimal solution pH for Ag(I) sorption is around 5.0. The sorption mechanism may include the chelation and redox reaction between Ag(I) ions and amine/imine groups on the PoPD chains. Similarly, the microparticles also have powerful Hg(II) adsorbability with 96.7% adsorptivity at an initial Hg(II) concentration of 4 mM. Competitive sorption between Ag(I) and Hg(II) in their mixture solution onto the microparticles was studied, exhibiting a preferential sorption toward Ag(I). The microparticles as a cost-effective sorbent demonstrate a promising application in the removal and even recovery of heavy-metal ions from wastewater. The PoPD/Ag nanocomposites possess (1) high Ag content of 34.8 wt %, (2) small diameter of Ag nanoparticles of around 10-20 nm, (3) narrow size distribution, (4) intrinsic electrical conductivity that is much higher than that of original PoPD microparticles without Ag. PMID- 19132886 TI - Mechanisms of composition change and toxic potentiation of chloramidophos emulsifiable concentrate during storage. AB - Storage instability is one of the serious problems that greatly restrict pesticide use. Routine checks on the composition and toxicity of 30% emulsifiable concentrates (EC) of chloramidophos (CP) during storage indicated that 78.6% of the active ingredient had decreased, whereas the anti-acetylcholinesterase (AChE) activity of the formulation was potentiated by 3.5 times. To understand the mechanism for these changes, detailed knowledge of the products present and their effects on anti-AChE potential deserves attention. It was likely that the basis for these changes was methanol, the cosolvent of CP EC, because when the purified CP was stored in methanol at 50 degrees C for 2 weeks, CP drop and toxic potentiation similar to those observed in CP EC also appeared. The major products of the CP-methanol reaction mixture were isolated and identified by HPLC and GC MS, respectively, and their inhibitory potentials against AChE and effectiveness as potentiators were assessed. Following redetermination of the main product (O,S dimethyl-[(2,2,2)-trichloro-1-methoxyethyl]phosphoramidothioate (MCP)) and high anti-AChE material (methamidophos), which were preconfirmed in the reaction mixture in CP EC, it was successfully demonstrated that the majority of CP in the formulation had been transformed to a new stable compound, MCP. Meanwhile, formation of another product, methamidophos, resulted in toxic potentiation. PMID- 19132887 TI - Using functional tissue engineering and bioreactors to mechanically stimulate tissue-engineered constructs. AB - Bioreactors precondition tissue-engineered constructs (TECs) to improve integrity and hopefully repair. In this paper, we use functional tissue engineering to suggest criteria for preconditioning TECs. Bioreactors should (1) control environment during mechanical stimulation; (2) stimulate multiple constructs with identical or individual waveforms; (3) deliver precise displacements, including those that mimic in vivo activities of daily living (ADLs); and (4) adjust displacement patterns based on reaction loads and biological activity. We apply these criteria to three bioreactors. We have placed a pneumatic stimulator in a conventional incubator and stretched four constructs in each of five silicone dishes. We have also programmed displacement-limited stimuli that replicate frequencies and peak in vivo patellar tendon (PT) strains. Cellular activity can be monitored from spent media. However, our design prevents direct TEC force measurement. We have improved TEC stiffness as well as PT repair stiffness and shown correlations between the two. We have also designed an incubator to fit within each of two electromagnetic stimulators. Each incubator provides cell viability like a commercial incubator. Multiple constructs are stimulated with precise displacements that can mimic ADL strain patterns and record individual forces. Future bioreactors could be further improved by controlling and measuring TEC displacements and forces to create more functional tissues for surgeons and their patients. PMID- 19132888 TI - Licorice flavonoids inhibit eotaxin-1 secretion by human fetal lung fibroblasts in vitro. AB - Glycyrrhiza uralensis (Gan-Cao), commonly called "licorice", is one of the most commonly used herbs in traditional Chinese medicine (TCM). In the United States, licorice products are most often consumed as flavoring and sweetening agents in food products. The licorice triterpenoid glycyrrhizin has several biological activities, including anti-inflammatory activity. Other potential anti inflammatory constituents in G. uralensis have not been fully investigated. Airway eosinophilic inflammation is a major feature of allergic asthma. Eotaxin-1 (eotaxin) is involved in the recruitment of eosinophils to sites of antigen induced inflammation in asthmatic airways. Because human lung fibroblasts are the major source of eotaxin, inhibition of eosinophil recruitment by suppression of fibroblast eotaxin production is a potentially valuable approach for the pharmacological intervention in asthma. A systematic bioassay-guided purification of G. uralensis yielded five flavonoids: liquiritin, liquiritigenin, isoliquiritigenin, 7,4'-dihydroxyflavone, and isoononin. The structures of the compounds were established by (1)H and (13)C nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS) studies. The potential ability of these isolated pure compounds and glycyrrhizin to inhibit secretion of eotaxin-1 by human fetal lung fibroblasts (HFL-1) was tested. Liquiritigenin, isoliquiritigenin, and 7,4'-dihydroxyflavone were more effective than liquiritin, isoononin, and glycyrrhizin in suppressing eotaxin secretion. A concentration response study showed the IC(50) concentrations of liquiritigenin, isoliquiritigenin, and 7,4'-dihydroxyflavone were 4.2, 0.92, and 0.21 microg/mL, respectively, demonstrating that Glycyrrhiza flavonoids inhibit eotaxin-1 secretion in vitro. PMID- 19132889 TI - Authentication of organic milk using delta13C and the alpha-linolenic acid content of milk fat. AB - The carbon stable isotope ratio (delta(13)C) and the alpha-linolenic acid (C18:3omega3) content of milk fat were analyzed to examine their applicability as general markers for the authentication of organic retail milk in Germany. To record the variable effect of feeding, including the seasonal influence on milk composition, three conventionally and three organically produced brands of retail milk were collected biweekly during a period of 18 months. Altogether 286 milk samples were analyzed. Threshold values for the identification of German organic milk were established and allowed to delimit almost all conventional samples. Organic retail milk was always above a minimum C18:3omega3 content of 0.50% and below a maximum delta(13)C of -26.5 per thousand. The universal and strongly negative correlation (r = -0.93) between C18:3omega3 and delta(13)C impedes the intentional manipulation of conventional milk. Conventional milk can naturally exceed the C18:3omega3 limit under atypical and rare conditions, but differentiation from organic milk can be improved by time-resolved comparison of data. In contrast with the general opinion, organic milk did not generally contain more c9,t11-C18:2 (CLA) than conventional milk. The proposed limits may deviate with dairy products containing milk from foreign countries. PMID- 19132890 TI - The effect of microstructure of octacalcium phosphate on the bone regenerative property. AB - The present study was designed to investigate whether the microstructure of synthetic octacalcium phosphate (OCP) affects its intrinsic bone regenerative properties as a scaffold and its conversion process into hydroxyapatite (HA). Our previous studies indicated that an agregate of OCP crystals, consisting of randomly oriented plate-like crystals, are capable of enhancing both osteoblastic cell differentiation in vitro and bone regeneration. While the transformation of OCP into HA has been considered in relation to the stimulatory capacity of OCP in bone regeneration, little is known about the effect of the microstructure of OCP granules on these capabilities. Two types of OCP granules, with identical diameters (300-500 microm) but composed of crystals with distinct crystal dimensions (4.0 and 26.6 microm length), were prepared (hereafter referred to as fine OCP granules [F-OCP] and coarse OCP granules [C-OCP], respectively). The intergranule distances and the porosity, including the intergranule spaces, were 108.5 microm and 93.7% for F-OCP, and 67.5 microm and 95.7% for C-OCP, as estimated by mercury intrusion. The OCP granules were implanted in mouse critical sized calvarial defects for up to 14 days. Histological examination demonstrated that osteoblastic cells aligned on the surface of F-OCP at day 7 and formed new bone around the granules up to day 14. On the other hand, cells around C-OCP were sparse at day 7, and resulted in only slight bone formation around the granules at day 14. X-ray diffraction showed that both OCP granules tended to be converted to an apatite structure with similar conversion velocity by the implantation. Adhesion of mouse bone marrow stromal ST-2 cells was markedly inhibited on C-OCP compared to F-OCP in vitro. These results suggested that the microstructure consisting of plate-like crystals of OCP controls cell adhesion on the crystal surfaces and their resultant bone regenerative properties as well as the physicochemical effect associated with the transitory nature of OCP previously reported. PMID- 19132891 TI - Scaffold-free cartilage fabrication system using passaged porcine chondrocytes and basic fibroblast growth factor. AB - The scaffold-free cartilage fabrication system we have reported previously for human application with nonpassaged cells has a big limitation in securing the enough cell number. Therefore, autologous chondrocytes from small biopsy of cartilage tissue inevitably have to be expanded through multiple passages, which result in poor engineered cartilage in terms of quality and quantity. This study applied basic fibroblast growth factor (bFGF) to overcome the limitation and produce an engineered cartilage tissue with clinically applicable size and quality. Porcine articular chondrocytes were expanded until passage 2 in the absence or presence of 5 ng/mL bFGF, and then subjected to the two-stage scaffold free cultures for 21 days again in the absence or presence of 5 ng/mL bFGF. The fabrication of cartilage tissues was evaluated along with time in comparison with the constructs from unpassaged chondrocytes. Expansion of chondrocytes in the presence of bFGF increased accumulation of cartilage extracellular matrices and resultantly fabricated the larger size of cartilage tissues in the subsequent stages, being comparable to those of unpassaged cells. In contrast, bFGF showed no positive, but adverse, effects on the cartilage tissue formation, when treated additionally during the scaffold-free fabrication stages. These results suggested that use of bFGF during the expansion stage of chondrocytes could overcome the limitation of previous two-stage scaffold-free cartilage fabrication system, and provided a novel three-stage system to construct a clinically applicable quality of cartilage tissues. PMID- 19132892 TI - Human neonatal thymus-derived mesenchymal stromal cells: characterization, differentiation, and immunomodulatory properties. AB - Mesenchymal stromal cells represent an attractive cell population for cell transplantation and tissue engineering purposes. The aim of this study was to search for neonatal thymus-derived mesenchymal stromal cells (nTMSC) and further characterize the differentiation and immunomodulatory properties thereof. The thymus glands of 13 infants undergoing congenital cardiac surgery were removed. After in vitro isolation and expansion, we identified adherent stromal cells with substantial proliferation potential. As characterized by FACS, the pattern of surface antigen expression of nTMSC resembled bone marrow stromal cells. Full mesenchymal differentiation potential is maintained during proliferation as confirmed by cultures for osteogenic, chondrogenic, and adipogenic lineages. After 5-azacytidine enrichment, morphological characteristics of cardiomyocytes were achieved. For immunologic investigations, the influence of nTMSC on the proliferative behavior of peripheral blood mononuclear cells was studied as a measure of the immune response. The nTMSC did not stimulate an allogeneic reaction in this coculture. Further, the expression of immunologically relevant markers was measured. Alike MSC from other origins, nTMSC did not express MHC-II. In contrast to mature MSC, some nTMSC even lack the expression of MHC-I. Our results confirm that the neonatal thymus contains mesenchymal stromal cells (nTMSC) with full mesenchymal differentiation potential and immunomodulatory properties. PMID- 19132893 TI - In vivo osteogenic differentiation of human adipose-derived stem cells in an injectable in situ-forming gel scaffold. AB - The sol-to-gel transition occurring at around body temperature makes the MPEG-PCL diblock copolymer an ideal candidate material for use as an injectable in situ forming gel containing human adipose tissue-derived stem cells (hADSCs). The sol can be prepared at room temperature, and the gel forms at body temperature. Solutions of the copolymer containing hADSCs and osteogenic factors injected into rats formed gel scaffolds at the injection sites. The gels thus formed showed the interconnective pore structure required to support growth, proliferation, and differentiation of hADSCs. Bromodeoxyuridine-labeled hADSCs were confirmed to be present in gels formed in vivo. Bone formation was observed only in gel implants containing both hADSCs and osteogenic factors. Subcutaneous implantation of the in situ-forming gel scaffold demonstrated that hADSCs embedded in the gel stimulated much lower host tissue responses than did the gel alone, probably because of the unique immunomodulatory properties of hADSCs. In conclusion, our data on hADSCs embedded in an in situ gel scaffold suggest that this formulation may provide numerous benefits as a noninvasive alternative for tissue-engineered bone formation. PMID- 19132894 TI - Production and characterization of a panel of monoclonal antibodies against native human cellular prion protein. AB - The human prion diseases, such as variant Creutzfeldt-Jakob disease (vCJD), are characterized by the conversion of the normal cellular prion protein (PrP(C)) into an abnormal disease associated form (PrP(Sc)). Monoclonal antibodies (MAbs) that recognize these different PrP isoforms are valuable reagents both in the diagnosis of these diseases and in prion disease research in general but we know of no attempts to raise MAbs against native human PrP(C). We immunized prion protein gene ablated (PrP(-/-)) mice with native human PrP(C) purified from platelets (pHuPrP) generating a predominantly IgG isotype anti-pHuPrP polyclonal antibody response in all mice. Following fusion of splenocytes from the immunized mice with SP2/0 myeloma cells, we were able to identify single cell clone and cryopreserve 14 stable hybridoma cell lines producing MAbs that reacted with pHuPrP. The properties of these MAbs (such as isotype, binding to native/denatured pHuPrP, and HuPrP epitopes recognized) are described. Furthermore, several of these MAbs showed a selectivity in their ability to immunoprecipitate disease associated PrP(Sc) and its corresponding protease resistant core (PrP(res)). PMID- 19132895 TI - Monoclonal antibody and siRNAs for topoisomerase I suppress telomerase activity. AB - Telomerase, a ribonucleoprotein enzyme, is a cellular RNA-dependent DNA polymerase that serves to maintain the tandem arrays of telomeric repeats at the eukaryotic chromosomal ends. We previously reported that topoisomerase I dissociates HIV-1 reverse transcriptase from genomic RNAs, and binding of topoisomerase I to RNA template regulates cDNA synthesis. We also found that a monoclonal antibody (MAb) against topoisomerase I, designated as MAb 1, suppresses the reverse transcription efficiency using a detergent-disrupted HIV-1 virion. In this study, we describe how MAb 1 suppresses telomerase activity in cellular lysates. In addition, siRNAs of topoisomerase I has attenuated telomerase activity in culture cells. These results suggest that topoisomerase I is involved in telomerase activity, as well as HIV-1 reverse transcription. PMID- 19132896 TI - Nuclear exclusion of p33ING1b tumor suppressor protein: explored in HCC cells using a new highly specific antibody. AB - Mouse monoclonal antibodies (MAb) were generated against p33ING1b tumor suppressor protein. 15B9 MAb was highly specific in recognizing a single protein band of approximately 33 kDa endogenous p33ING1b protein from HCC cell lines and normal liver tissue by Western blot analysis and by immunoprecipitation. Although p33ING1b mutations are rarely observed in cancer, differential subcellular distribution and nuclear exclusion of p33ING1b were reported in different cancer types. Therefore we analyzed the expression and subcellular localization of p33ING1b in HCC cell lines using 15B9 MAb. So far, p33ING1b mutations or differential subcellular localization are not reported in HCC. In this study, by indirect immunofluorescence using MAb 15B9, we demonstrate that nuclear localization of p33ING1b was highly correlated with well-differentiated HCC cell lines whereas poorly differentiated HCC cells have nuclear exclusion of the protein. Moreover no association was observed between differential subcellular localization of p33ING1b and p53 mutation status of HCC cell lines. Hence our newly produced MAb 15B9 can be used for studying cellular activities of p33ING1b under normal and cancerous conditions. PMID- 19132897 TI - Lactimidomycin, iso-migrastatin and related glutarimide-containing 12-membered macrolides are extremely potent inhibitors of cell migration. AB - Migrastatin (1), iso-migrastatin (5) and lactimidomycin (7) are all glutarimide containing polyketides known for their unique structures and cytotoxic activities against human cancer cell lines. Migrastatin, a strong inhibitor of tumor cell migration, has been an important lead in the development of antimetastatic agents. Yet studies of the related 12-membered macrolides iso-migrastatin, lactimidomycin, and related analogues have been hampered by their limited availability. We report here the production, isolation, structural characterization, and biological activities of iso-migrastatin, lactimidomycin, and 23 related congeners. Our studies showed that, as a family, the glutarimide containing 12-membered macrolides are extremely potent cell migration inhibitors with some members displaying activity on par or superior to that of migrastatin as exemplified by compounds 5, 7, and 9-12. On the basis of these findings, the structures and activity of this family of compounds as cell migration inhibitors are discussed. PMID- 19132898 TI - Cooperative DNA probing using a beta-cyclodextrin-DNA conjugate and a nucleobase specific fluorescent ligand. AB - A single nucleotide polymorphism (SNP) base on the target is displayed at a gap in a ternary duplex carrying beta-cyclodextrin-modified DNA. A stable tandem duplex forms regardless of the type of SNP base. A nucleobase-specific ligand is then added to this system. The dansyl moiety in the ligand is expected to form a luminous inclusion complex with nearby beta-CyD, only when the ligand recognizes the specific base displayed in the gap. PMID- 19132899 TI - Direct cross-coupling reaction of simple alkenes with acrylates catalyzed by palladium catalyst. AB - An efficient methodology for oxidative cross-coupling reaction of olefins with acrylates catalyzed by palladium(II) was developed. The corresponding dienoates were obtained in moderate to good yields. PMID- 19132911 TI - Can blogging enhance subjective well-being through self-disclosure? AB - Based on the self-disclosure theory and the social capital theory, this study investigates if bloggers' self-disclosure enhances their social capital and if these capitals in turn enhance perception of subjective well-being (SWB). The results reveal that the self-disclosure of bloggers significantly and directly affects a blogger's perception of social integration, bonding social capital, and bridge social capital, which in turn promote bloggers' SWB. It appears that as bloggers share their inner thoughts of their moods/feelings with others through writing, they may gain greater social support and improve their social integration. Therefore, self-disclosure through blogging may serve as the core of building intimate relationships. Furthermore, social capital, built through blogging, may improve a blogger's satisfaction with his or her social contact, interpersonal communication, and overall quality of life. PMID- 19132912 TI - Behavioral indications of object-presence in haptic virtual environments. AB - The success of many virtual environment (VE) applications relies on their ability to induce in their users a sense of presence. In immersive VE, presence is a sense of being and acting inside a virtual place, whereas in a nonimmersive haptic VE, it is a sense of being able to touch and manipulate a virtual object. This latter sense of object-presence is typically measured by questionnaires, and the current study aimed to find objective behavioral indications for it. Participants moved a stylus along the blades of a virtual razor and along identical virtual lines with haptic feedback but without the context of a razor. Our measurements show that participants' movements were slower and they exerted less force on the stylus in the razor condition than in the simple lines condition. This behavioral pattern suggests that some degree of object-presence illusion was formed, which caused participants to act more cautiously in order to avoid any harm from the virtual razor. PMID- 19132913 TI - Agency, the sense of presence, and schizophrenia. AB - The objective of this work is to evaluate the impact of agency deficiency on presence. We hypothesize that a deficit in agency, such as occurs in schizophrenia, has repercussions on the sense of presence as well as on performance in a sensorimotor test involving the body. Nineteen healthy controls and 19 stable patients with schizophrenia, matched by age and gender, participated in the study. We used new interactive technologies that enable the users to physically interact with a virtual environment and simultaneously sees themselves performing the actions. Two conditions were balanced: the control condition in which the participant see himself or herself doing the test, and the mask condition, in which the central part of the participant's body is hidden. At the end of the test, all participants completed a questionnaire on presence. The results show that control participants performed better than those with schizophrenia in the control condition. On the contrary, in the mask condition, their performance was significantly lower, whereas participants with schizophrenia obtained the same score as in the control condition. Controls also rated higher in the scale of presence than did participants with schizophrenia. Those results seem to corroborate a relationship between agency and presence. The body awareness and the self in schizophrenia are discussed in the light of agency. PMID- 19132914 TI - Alexithymia and its relationships with dissociative experiences and Internet addiction in a nonclinical sample. AB - The aim of the present study was to evaluate alexithymia, dissociative experiences, and Internet addiction (IA) in a nonclinical sample of 312 undergraduate students, identifying predictive factors associated with the possible risk of developing IA. We found that alexithymics had more consistent dissociative experiences, lower self-esteem, and higher obsessive-compulsive symptoms than nonalexithymics. In addition, alexithymics reported a higher potential risk for IA when compared to nonalexithymics. Difficulty in identifying feelings, higher dissociative experiences, lower self-esteem, and higher impulse dysregulation were associated with higher IA. Thus, a combination of alexithymia, dissociative experiences, low self-esteem, and impulse dysregulation may be a risk factor for IA, at least in a nonclinical sample. PMID- 19132915 TI - IL-17 and Th17 Cells. AB - CD4+ T cells, upon activation and expansion, develop into different T helper cell subsets with different cytokine profiles and distinct effector functions. Until recently, T cells were divided into Th1 or Th2 cells, depending on the cytokines they produce. A third subset of IL-17-producing effector T helper cells, called Th17 cells, has now been discovered and characterized. Here, we summarize the current information on the differentiation and effector functions of the Th17 lineage. Th17 cells produce IL-17, IL-17F, and IL-22, thereby inducing a massive tissue reaction owing to the broad distribution of the IL-17 and IL-22 receptors. Th17 cells also secrete IL-21 to communicate with the cells of the immune system. The differentiation factors (TGF-beta plus IL-6 or IL-21), the growth and stabilization factor (IL-23), and the transcription factors (STAT3, RORgammat, and RORalpha) involved in the development of Th17 cells have just been identified. The participation of TGF-beta in the differentiation of Th17 cells places the Th17 lineage in close relationship with CD4+CD25+Foxp3+ regulatory T cells (Tregs), as TGF-beta also induces differentiation of naive T cells into Foxp3+ Tregs in the peripheral immune compartment. The investigation of the differentiation, effector function, and regulation of Th17 cells has opened up a new framework for understanding T cell differentiation. Furthermore, we now appreciate the importance of Th17 cells in clearing pathogens during host defense reactions and in inducing tissue inflammation in autoimmune disease. PMID- 19132917 TI - Blood monocytes: development, heterogeneity, and relationship with dendritic cells. AB - Monocytes are circulating blood leukocytes that play important roles in the inflammatory response, which is essential for the innate response to pathogens. But inflammation and monocytes are also involved in the pathogenesis of inflammatory diseases, including atherosclerosis. In adult mice, monocytes originate in the bone marrow in a Csf-1R (MCSF-R, CD115)-dependent manner from a hematopoietic precursor common for monocytes and several subsets of macrophages and dendritic cells (DCs). Monocyte heterogeneity has long been recognized, but in recent years investigators have identified three functional subsets of human monocytes and two subsets of mouse monocytes that exert specific roles in homeostasis and inflammation in vivo, reminiscent of those of the previously described classically and alternatively activated macrophages. Functional characterization of monocytes is in progress in humans and rodents and will provide a better understanding of the pathophysiology of inflammation. PMID- 19132918 TI - Chondrogenesis of adipose-derived adult stem cells in a poly-lactide-co-glycolide scaffold. AB - Adult adipose-derived stem cells (ASCs) are considered to be an alternative cell source for cell-based cartilage repair because of their multiple differentiation potentials. This article addresses the chondrogenic differentiation of ASCs seeded into poly-lactide-co-glycolide (PLGA) scaffolds after implantation in a subcutaneous pocket of nude mice. Human ASCs were seeded into PLGA (polylactic acid:polyglycolic acid = 90:10) scaffolds and cultured in transforming growth factor beta 1 (TGF-beta1)-containing medium for 3 weeks in vitro. Then specimens were implanted into a subcutaneous pocket of severe combined immunodeficiency mice and harvested after 8 weeks. Chondrospecific messenger RNA (mRNA) expression was analyzed using reverse transcriptase polymerase chain reaction. Corresponding extracellular matrix (ECM) synthesis was demonstrated using immunohistochemical staining. Chondrospecific marker molecules such as collagen type II and type X, cartilage oligomeric matrix protein, and aggrecan subsequently increased during the 3 weeks period in vitro. After a further 8 weeks, in vivo samples pretreated with TGF-beta1 continued expressing collagen type II and aggrecan mRNA, and collagen type II was found within the ECM using immunohistochemistry. Chondrospecific mRNA was not detected in control samples. ASC-seeded PLGA scaffolds express a stable chondrogenic phenotype in a heterotopic model of cartilage transplantation and represent a suitable tool for tissue engineering of cartilage. PMID- 19132919 TI - Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines. AB - Long-term in vitro culture of undifferentiated human embryonic stem cells (hESCs) traditionally requires a fibroblast feeder cell layer. Using feeder cells in hESC cultures is highly laborious and limits large-scale hESC production for potential application in regenerative medicine. Replacing feeder cells with defined human extracellular matrix (ECM) components or synthetic biomaterials would be ideal for large-scale production of clinical-grade hESCs. We tested and compared different feeder cell-free hESC culture methods based on different human ECM proteins, human and animal sera matrices, and a Matrigel matrix. Also selected biomaterials were tested for feeder cell-free propagation of undifferentiated hESCs. The matrices were tested together with conventional and modified hESC culture media, human foreskin fibroblast-conditioned culture medium, chemically defined medium, TeSR1, and modified TeSR1 media. The results showed the undefined, xenogeneic Matrigel to be a superior matrix for hESC culture compared with the purified human ECM proteins, serum matrices, and the biomaterials tested. A long-term, feeder cell-free culture system was successful on Matrigel in combination with mTeSR1 culture medium, but a xeno-free, fully defined, and reproducible feeder cell-free hESC culture method still remains to be developed. PMID- 19132920 TI - Spectral simulation protocol for extending the lifetime of near-infrared multivariate calibrations. AB - Multivariate calibration models based on synthetic single-beam near-infrared spectra are used to demonstrate the ability to maintain viable calibrations over extended time periods. Glucose is studied over the physiological concentration range of 1-30 mM in a buffered aqueous matrix containing varying levels of alanine, ascorbate, lactate, urea, and triacetin. By employing a set of 25 test samples measured 23 times over a period of 325 days, partial least-squares (PLS) calibrations based on synthetic spectra are observed to outperform conventional calibrations that use a set of 64 measured calibration samples. The key to the success of this approach is the use of a set of spectra of phosphate buffer collected on each prediction day to construct synthetic calibration spectra that are specific to that day. This allows the incorporation into the calibration model of nonanalyte spectral variance that is unique to a particular day. In this way, the adverse effects of instrumental drift or other sources of spectral variance on prediction performance can be minimized. Through the application of this methodology, values of the standard error of prediction (SEP) for glucose concentration are maintained to a range of 0.50-0.95 mM and an average of 0.68 mM over the 325 days of the experiment. These results are significantly better than those obtained with conventional models based on measured calibration samples. Over the same time period, a PLS model based on measured calibration spectra in absorbance units produced values of SEP that ranged from 0.41 to 2.02 mM and an average of 1.23 mM. PMID- 19132921 TI - Facilitation of applicability in ice chromatography by mechanistic considerations and by preparation of fine water-ice stationary phase. AB - We have, in previous papers, demonstrated that some solute is adsorbed on the ice surface at temperatures below -5 degrees C by the hydrogen-bond formation between the polar groups in the solute and the dangling bonds on the water-ice surface. However, the separation efficiency of this method, named ice chromatography, was seriously restricted principally due to the large sizes of water-ice particles used as a stationary phase. We have devised a convenient method to prepare finer ice particles with diameters of approximately 10 microm. This stationary phase has provided much larger theoretical plate numbers (N = 1500) than that prepared in the previous way (N = 250) and has allowed various applications such as separation of amino acid derivatives, poly(oxyethylene) oligomers, and estrogen. The improved separation performance allows us to discuss retention mechanisms in more detail and to see subtle differences in the retentivity between solutes. PMID- 19132916 TI - T cell activation. AB - This year marks the 25th anniversary of the first Annual Review of Immunology article to describe features of the T cell antigen receptor (TCR). In celebration of this anniversary, we begin with a brief introduction outlining the chronology of the earliest studies that established the basic paradigm for how the engaged TCR transduces its signals. This review continues with a description of the current state of our understanding of TCR signaling, as well as a summary of recent findings examining other key aspects of T cell activation, including cross talk between the TCR and integrins, the role of costimulatory molecules, and how signals may negatively regulate T cell function.Acronyms and DefinitionsAdapter protein: cellular protein that functions to bridge molecular interactions via characteristic domains able to mediate protein/protein or protein/lipid interactions Costimulation: signals delivered to T cells by cell surface receptors other than the TCR itself that potentiate T cell activation cSMAC: central supramolecular activation cluster Immunoreceptor tyrosine-based activation motif (ITAM): a short peptide sequence in the cytoplasmic tails of key surface receptors on hematopoietic cells that is characterized by tyrosine residues that are phosphorylated by Src family PTKs, enabling the ITAM to recruit activated Syk family kinases Inside-out signaling: signals initiated by engagement of immunoreceptors that lead to conformational changes and clustering of integrins, thereby increasing the affinity and avidity of the integrins for their ligands NFAT: nuclear factor of activated T cells PI3K: phosphoinositide 3 kinase PKC: protein kinase C PLC: phospholipase C pMHC: peptide major histocompatibility complex (MHC) complex pSMAC: peripheral supramolecular activation cluster PTK: protein tyrosine kinase Signal transduction: biochemical events linking surface receptor engagement to cellular responses TCR: T cell antigen receptor PMID- 19132922 TI - Oxidation and glycolytic cleavage of etheno and propano DNA base adducts. AB - Non-invasive strategies for the analysis of endogenous DNA damage are of interest for the purpose of monitoring genomic exposure to biologically produced chemicals. We have focused our research on the biological processing of DNA adducts and how this may impact the observed products in biological matrixes. Preliminary research has revealed that pyrimidopurinone DNA adducts are subject to enzymatic oxidation in vitro and in vivo and that base adducts are better substrates for oxidation than the corresponding 2'-deoxynucleosides. We tested the possibility that structurally similar exocyclic base adducts may be good candidates for enzymatic oxidation in vitro. We investigated the in vitro oxidation of several endogenously occurring etheno adducts [1,N(2)-epsilon guanine (1,N(2)-epsilon-Gua), N(2),3-epsilon-Gua, heptanone-1,N(2)-epsilon-Gua, 1,N(6)-epsilon-adenine (1,N(6)-epsilon-Ade), and 3,N(4)-epsilon-cytosine (3,N(4) epsilon-Cyt)] and their corresponding 2'-deoxynucleosides. Both 1,N(2)-epsilon Gua and heptanone-1,N(2)-epsilon-Gua were substrates for enzymatic oxidation in rat liver cytosol; heteronuclear NMR experiments revealed that oxidation occurred on the imidazole ring of each substrate. In contrast, the partially or fully saturated pyrimidopurinone analogues [i.e., 5,6-dihydro-M(1)G and 1,N(2) propanoguanine (PGua)] and their 2'-deoxynucleoside derivatives were not oxidized. The 2'-deoxynucleoside adducts, 1,N(2)-epsilon-dG and 1,N(6)-epsilon dA, underwent glycolytic cleavage in rat liver cytosol. Together, these data suggest that multiple exocyclic adducts undergo oxidation and glycolytic cleavage in vitro in rat liver cytosol, in some instances in succession. These multiple pathways of biotransformation produce an array of products. Thus, the biotransformation of exocyclic adducts may lead to an additional class of biomarkers suitable for use in animal and human studies. PMID- 19132923 TI - Integration host factor (IHF) dictates the structure of polyamine-DNA condensates: implications for the role of IHF in the compaction of bacterial chromatin. AB - Integration host factor (IHF), a nucleoid-associated protein in bacterial cells, is implicated in a number of chromosomal functions including DNA compaction. IHF binds to all duplex DNA with micromolar affinity and at sequence-specific sites with much higher affinity. IHF is known to induce sharp bends in the helical axis of DNA in both modes of binding, but the role of IHF in controlling DNA condensation within bacterial cells has remained undetermined. Here we demonstrate that IHF influences the morphology of DNA condensed by polyamines in vitro. In the absence of IHF, spermidine and spermine condense DNA primarily into toroidal structures, whereas in the presence of IHF, polyamines condense DNA primarily into rodlike structures. Computer simulations of DNA condensation in the absence and presence of IHF binding lend support to our model in which DNA bending proteins, such as IHF and HU, promote the condensation of DNA into rodlike structures by providing the free energy necessary to bend DNA at the ends of linear bundles of condensed DNA. We propose that a common function of IHF and HU in bacterial cells is to facilitate DNA organization in the nucleoid by the introduction of sharp bends in chromosomal DNA. PMID- 19132924 TI - Exploring a series of isostructural dodecanuclear mixed Ni:Co clusters: toward the control of elemental composition using pH and stoichiometry. AB - The compositional parameter space in the formation of polynuclear clusters is probed in the synthesis of a series of dodecanuclear coordination clusters of Ni(II) and Co(II) with isostructural D(3h)-symmetric frameworks. At the core of their construction are a carbonate template and the directing ligands cis,trans 1,3,5-triaminocyclohexane and acetate at contrasting pH values. The pH and stoichiometric dependence has been mapped, and analysis by electrospray mass spectrometry reveals the cluster cores in solution. In two specific cases, site specific occupations are eluded to by analysis of the magnetic properties, and we discuss the possibility of controlling the molecular composition of mixed metal polynuclear clusters. PMID- 19132925 TI - Preparation of bicyclic 1,2,4-trioxanes from gamma,delta-unsaturated ketones. AB - Treatment of gamma,delta-unsaturated ketones with hydrogen peroxide and acid provides a rapid entry into the medicinally important 1,2,4-trioxane structure. Alkene substitution that stabilizes carbocationic intermediates proved to be important for the success of this transformation. PMID- 19132926 TI - An approach to 3,6-disubstituted 2,5-dioxybenzoquinones via two sequential Suzuki couplings. three-step synthesis of leucomelone. AB - Two sequential Suzuki coupling reactions have been developed for efficient synthesis of synthetically and biologically important 3,6-disubstituted 2,5 dioxybenzoquinone architectures in a highly chemoselective controlled manner. The method serves as a key step in the total synthesis of leucomelone in three steps and in 61% overall yield. PMID- 19132927 TI - Measurement of ring A-reduced progesterone metabolites by enzyme-linked immunoassay with colorimetric detection: baseline levels of six metabolites, including pregnanolone, in male rat plasma. AB - The performance of an antiserum to progesterone and pregnane neurosteroids was assessed in two competitive assay setups: radioimmunoassay and enzyme-linked immunoassay with colorimetric detection, both with the same limit of detection of 2 pg. The enzyme-linked immunoassay was less labor-intensive and had better precision of measurement and was used to measure progesterone and six of its ring A-reduced metabolites in rat plasma. The measured levels of allopregnanolone and progesterone were in agreement with those reported previously when measured by gas chromatography/mass spectrometry and high-performance liquid chromatography coupled with radioimmunoassay and substantially lower than those previously measured by radioimmunoassay without chromatographic separation. Both isomers of dihydroprogesterone and all four isomers of pregnanolone were detected in rat plasma, indicating that progesterone is metabolized by reduction at the C5 and C3 position of the A ring, in both alpha and beta configurations. In addition to 5beta-dihydroprogesterone and isopregnanolone, which have not been previously detected in the rat, we found considerable amounts of pregnanolone, which is neuroactive, with similar potency to that of allopregnanolone but was previously thought not to be produced in rats. PMID- 19132928 TI - From proteome to genome for functional characterization of pH-dependent outer membrane proteins in Escherichia coli. AB - Proteomic technology is very powerful in identification of differentially expressed proteins. However, how to identify key proteins and distinguish them from others has been a question to be solved in functional proteomics. Utilizing 2-D gel based proteomic approach, we identified 11 differentially expressed outer membrane (OM) proteins involved in E. coli's response to pH change. The protein expression changes were validated by Western blotting. The function and roles of the differentially expressed proteins were further characterized using genetically modified strains with gene deletion of these altered OM proteins and gene complementation or overexpression approach. Among the differentially expressed proteins identified, OstA, TolC, OmpT, OmpP OmpC, Trak, OmpX, Dps, LamB, Tsx, FadL, OmpW, and OmpF were characterized as pH-related OM proteins. Out of these OM proteins, TolC, OmpC, OmpX, and LamB may play critical roles in pH regulation in E. coli. Using death-rescuing assay developed in house, we found that OmpC, LamB, FadL, OmpX, OmpW, and OmpF, LamB, FadL, and OmpW functioned in a TolC-independent pathway, whereas OmpF, Tsx and OmpC, OmpX, and Tsx might share the same pathway with TolC at the extreme acid or base condition. The information obtained from this study provides novel insight into mechanisms of pH response in E. coli. Our results also demonstrate the importance and efficiency of functional characterization of differentially expressed proteins at different molecular levels in identification of key target proteins and pathways involved in E. coli' s response to pH change. PMID- 19132929 TI - The use of drug metabolism for prediction of intestinal permeability (dagger). AB - The Biopharmaceutics Classification System (BCS), based on the aqueous solubility and intestinal permeability of a drug substance, has been widely used to predict the extent of drug absorption during the course of pharmaceutical development. Combined with product dissolution data, this system has gained a prominent role in regulatory process to determine if a drug formulated in an immediate release solid oral dosage form qualifies for waiver of in vivo bioequivalence studies. In parallel, the Biopharmaceutics Drug Disposition Classification System (BDDCS), using aqueous solubility and drug metabolism, takes on another venue to predict overall drug disposition. It has been suggested that the matrix of drug metabolism in BDDCS can be used to substantiate the classification of permeability by BCS. A total of 51 drugs were compiled in this study to examine the use of drug metabolism for predicting permeability. All compounds were classified as high permeability based on BCS, but only 73% of the compounds were found to exhibit extensive metabolism. Lipophilicity accounts for significant metabolism of many highly permeable drugs. Fourteen (14) out of 51 drugs have poor metabolism, suggesting that high permeability as defined by BCS does not necessarily dictate extensive metabolism. The drugs that have high permeability but poor metabolism are generally hydrophilic molecules with low molecular weight and are likely to be absorbed by active transport mechanisms. Based on the present data and literature information, it seems logical to predict that the extent of absorption is mostly complete (or > or =90%) if the drug is subject to a high degree of metabolism (e.g., > or =90%). The extent of drug metabolism may be useful in supporting permeability classification under certain circumstances. PMID- 19132930 TI - Synthesis of length-controlled aerosol carbon nanotubes and their dispersion stability in aqueous solution. AB - A one-step method combining spray pyrolysis and thermal chemical vapor deposition (CVD) processes was developed to grow hybrid carbon nanotube (CNT)-bimetallic composite particles. Nickel, aluminum, and acetylene were used as the catalytic site, noncatalytic matrix, and hydrocarbon source, respectively. The bimetallic particles (i.e., Al-Ni) were spray pyrolized and subsequently passed through thermal CVD. During the thermal CVD, the catalytic decomposition of acetylene occurred on the free-floating bimetallic particles so that sea urchin-like CNTs were radially grown. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses revealed the CNTs to have a uniform diameter of approximately 10 +/- 2 nm. The length of the CNTs was controlled by varying the residence time of the bimetallic nanoparticles with a length of 200-1000 nm. After nitric acid treatment, the CNTs were released by melting the bimetallic particles. The resulting CNTs were then dispersed in an aqueous solution to examine the effect of the length of CNTs on their dispersion stability, which is a critical issue for the stability and repeatability of the heat transfer performance in nanofluids. Ultraviolet-visible (UV-vis) spectrometer analysis showed that shorter CNTs were less stable than the longer CNTs due to the higher mobility-induced agglomeration of the shorter CNTs. PMID- 19132931 TI - New insights into the torquoselectivity of the Staudinger reaction. AB - To understand the torquoselectivity and the electronic effects of the Staudinger reaction, a study using a combination of experiments and DFT calculations has been conducted for the reactions of an unsymmetric cyclic ketene and cyclic imines with different electronic properties. The predominant formation of the donor-in beta-lactams, the torquoelectronically disfavored products, was observed for the first time. The Hammett analyses reveal a close relationship between the donor-out/donor-in product ratio and the electronic nature of the imines. The kinetic competition experiments and DFT calculations indicate that the two-step Staudinger reaction shows different rate-determining steps in the donor-in and donor-out pathways. Our investigations reveal that the torquoelectronic control in the Staudinger reaction is quite different from that in the ring-opening reaction of cyclobutene derivatives. The diastereoselectivity of the Staudinger reaction involving an unsymmetric disubstituted ketene cannot be simply rationalized or predicted by the torquoelectronic model. PMID- 19132932 TI - Effects of hyperconjugation on the electronic structure and photoreactivity of organic sulfonyl chlorides. AB - The electronic structure of organic sulfonyl compounds of the form RSO(2)G (G = Cl, -OH, -CH(3)) is investigated to evaluate the effect of aryl R groups on photocleavage of the S-G bond. Sulfur K-edge X-ray absorption spectroscopy (XAS) provides a direct measure of the empty low-lying molecular orbitals in these complexes and, in combination with DFT calculations, a detailed description of the bonding in these compounds. The presence of an aryl group bound to the sulfonyl moiety has a significant impact on the spectroscopy and electronic structure of the site. The analysis suggests that the SCl(sigma*) orbital is significantly affected by mixing with the aryl pi* manifold. This mixing is dependent upon the nature of G and is most pronounced in the sulfonyl chlorides, where the energy of the SCl(sigma*) orbital is lowered by approximately 0.5 eV. The observed mixing is best described as excited-state hyperconjugation of the aryl pi system into the SCl(sigma*) orbital. The magnitude of the effect can be estimated directly from the S K-edge XAS spectra. These results are discussed in relation to the observed photochemistry of RSO(2)Cl, which is significantly enhanced when R = aryl as compared to alkyl substituents. PMID- 19132934 TI - Melanin synthesis inhibitors from Lespedeza floribunda. AB - In the course of our search for new melanin synthesis inhibitors from plants, 40 new flavonoids and 11 known flavonoids were isolated from the roots of Lespedeza floribunda Bunge. The structures of the new compounds were determined by MS and NMR analyses, and the absolute configurations by CD spectra. Many of the compounds inhibited melanin synthesis in normal human epidermal melanocytes (NHEM), and compounds 3, 7, 8, 11, 16, 24, 27, 29, 33, 43, 45, and 51 were particularly inhibitory. Their activities were stronger than that of hydroquinone, which is known as a major skin-lightening drug. PMID- 19132933 TI - Generation of DNA interstrand cross-links by post-synthetic reductive amination. AB - DNA interstrand cross-links (ICLs) are the clinically most relevant adducts formed by many antitumor agents. To facilitate the study of biological responses triggered by ICLs, we developed a new approach toward the synthesis of mimics of nitrogen mustard ICLs. 7-Deazaguanine residues bearing acetaldehyde groups were incorporated into complementary strands of DNA and cross-link formation induced by double reductive amination. Our strategy enables the synthesis of major groove cross-links in high yields and purity. PMID- 19132935 TI - Antibiofilm activity of a diverse oroidin library generated through reductive acylation. AB - A diverse 20-compound library of analogues based on the marine alkaloid oroidin were synthesized via a reductive acylation strategy. The final target was then assayed for inhibition and dispersion activity against common proteobacteria known to form biofilms. This methodology represents a significant improvement over the generality of known methods to acylate substrates containing 2 aminoimidazoles and has the potential to have broad application to the synthesis of more advanced oroidin family members and their corresponding analogues. PMID- 19132936 TI - Efficient access to a versatile 5,6-dithio-1,10-phenanthroline building block and corresponding organometallic complexes. AB - A facile access to 5,6-bis(2-cyanoethylsulfanyl)-1,10-phenanthroline 1 and its ruthenium(II) bipyridil complex 2, as versatile building blocks for the straightforward synthesis of 5,6-dithio functionalized 1,10-phenanthroline based systems, is described. PMID- 19132937 TI - Oxidative cyclization reactions of trienes and dienynes: total synthesis of membrarollin. AB - Trienes and dienynes containing one electron-deficient double bond were shown to undergo regio- and stereoselective oxidative cyclization in the presence of permanganate ion to afford 2,5-bis-hydroxyalkyltetrahydrofurans (THF diols). The THF diols produced retained either alkene or alkyne functionalities, which provided convenient handles for the metal oxo-mediated introduction of an adjacent THF ring with overall control of relative and absolute stereochemistry. Adjacent bis-THFs possessing threo-cis-threo-trans-erythro, threo-cis-threo-trans threo, threo-cis-threo-cis-erythro, threo-cis-erythro-cis-threo, or threo-cis erythro-trans-threo relationships were synthesized by appropriate selection of alkene geometry and methodology for the closure of the second ring. The threo-cis threo-cis-erythro stereochemical arrangement is embodied within the bis-THF core units of a number of Annonaceous acetogenins including membrarollin, while trilobacin has a threo-cis-erythro-trans-threo configured core. As an application of the selective oxidative cyclization approach, a total synthesis of membrarollin was completed in 17 linear steps from dodecyne. The C21,C22 double epimer of membrarollin was also synthesized in 15 linear steps and without recourse to the use of hydroxyl group protection. PMID- 19132938 TI - Lotus-like biomimetic hierarchical structures developed by the self-assembly of tubular plant waxes. AB - Hierarchical roughness is beneficial for superhydrophobic and self-cleaning surfaces. Biomimetic hierarchical surfaces were fabricated by replication of a micropatterned master surface and self-assembly of two kinds of tubular wax crystals, which naturally occur on the superhydrophobic leaves of Tropaeolum majus (L.) and Leymus arenarius (L.). These tubule forming waxes are multicomponent waxes, composed of a mixture of long chain hydrocarbons. Thermal evaporation of wax was used to cover artificial surfaces with a homogeneous wax layer and tubule formation was initiated by temperature and a solvent vapor phase. Based on this technique, various nanostructures produced by three dimensional tubular waxes have been fabricated by changing the wax mass. Fabricated structures and surface chemistry mimic the hierarchical surfaces of superhydrophobic and self-cleaning plant surfaces. The influence of structures on superhydrophobicity at different length scales is demonstrated by investigation of contact angle, contact angle hysteresis, droplet evaporation and propensity of air pocket formation as well as adhesive forces. The optimal structural parameters for superhydrophobicity and low static contact angle hysteresis, superior to natural plant leaves including Lotus, have been identified and provide a useful guide for development of biomimtetic superhydrophobic surfaces. PMID- 19132939 TI - Amide-directed alkenylation of sp(2) C-H Bonds catalyzed by a cationic Rh(I)/BIPHEP complex under mild conditions: dramatic rate acceleration by a 1 pyrrolidinecarbonyl group. AB - A cationic rhodium(I)/BIPHEP complex catalyzes amide-directed regioselective alkenylations of olefinic or aromatic sp(2) C-H bonds in good yields under mild reaction conditions. The use of a 1-pyrrolidinecarbonyl group as a directing group dramatically accelerates the reaction. PMID- 19132940 TI - Excited state carbene formation from UV irradiated diazomethane. AB - The laser flash photolysis process of diazomethane has been studied by using a real time propagation time-dependent density functional theory (RTP-TDDFT) combined with molecular dynamics. The activation energy barrier for disintegrating diazomethane into nitrogen (N(2)) and carbene (CH(2)) molecules significantly decreases in the electronic excited S(1) state compared to that in the S(0) ground state. Furthermore, the produced carbene molecule can be in the electronic excited state of (1)CH(2) ((1)B(1)) instead of the lowest state among singlet states (1)CH(2) ((1)A(1)), which is evident in the wave function characteristics of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) throughout the disintegration. This is regarded as the initial stage of the rearrangement in the excited state (RIES), the evidence of which has been given by experiments in the past decade. In the RIES mechanism scheme, we suggest that the photoreaction in the S(1) state contributes considerably to the photochemistry of carbene formation. The passing near the S(1)/S(0) conical intersection, which allows the transition to ground state diazomethane producing the lowest singlet state carbene molecule, is considered a rare event from our molecular dynamics, although this has been regarded as the dominant mechanism in previous theoretical studies. PMID- 19132941 TI - Concise and efficient synthesis of 2-acetamido-2-deoxy-beta-D-hexopyranosides of diverse aminosugars from 2-acetamido-2-deoxy-beta-D-glucose. AB - The furanose acetonide derivative 1 is readily prepared from 2-acetamido-2-deoxy D-glucose on a large scale without the need for chromatography. Mesylation of 1 provides an efficient, concise, synthetic route to rare 2-acetamido-2-deoxy-beta D-hexopyranosides (2 and 3) via the corresponding methyl 2-acetamido-2-deoxy-3-O methanesulfonyl-beta-D-glucopyranoside and subsequent inversion of configuration by direct displacement or formation of a 3,4-epoxide. Opening of this epoxide by azide provided a direct route to methyl 2-acetamido-4-amino-2,4,6-trideoxy-beta-D gulopyranoside 4. Benzylation of 1 followed by ring expansion to the glucopyranoside, deoxygenation at C-6, and subsequent displacement of a C-4 triflate permitted the synthesis of methyl 2-acetamido-4-amino-2,4,6-trideoxy beta-D-galactopyranoside 5. Methyl 2-acetamido-2-deoxy-beta-D-glucopyranoside available from 1 in quantitative yield was readily converted to methyl 2 acetamido-2-deoxy-beta-D-galactopyranoside 6 (>60%) by inversion of configuration at C-4. Introduction of a lactyl substituent at C-3 of oxazoline 1 also provides a facile synthesis of the biologically important muramic acid beta-glycoside 7. An interesting reaction to convert 2-acetamido-2-deoxyhexopyranosides to the corresponding 2-deoxy-2-tetrazole is also reported. PMID- 19132942 TI - Synthesis of phenol and quinone metabolites of benzo[a]pyrene, a carcinogenic component of tobacco smoke implicated in lung cancer. AB - Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental pollutants produced in the combustion of organic matter. PAHs are present in automobile exhaust and tobacco smoke, and they have recently been designated as human carcinogens. Current evidence indicates that PAHs are activated enzymatically to mutagenic metabolites that interact with DNA. There is evidence for three pathways of activation: the diol epoxide path, the radical-cation path, and the quinone path. The relative importance of these paths for human lung cancer has not been established. We now report syntheses of the principal phenol and quinone isomers of the prototype PAH carcinogen benzo[a]pyrene (BP) that are known or are suspected to be formed as metabolites of BP in human bronchoalveolar cells. The methods of synthesis were designed to be adaptable to the preparation of the (13)C-labeled analogues of the BP metabolites. These compounds are needed as standards for sensitive LC-MS/MS methods for analysis of BP metabolites formed in lung cells. Efficient novel syntheses of the 1-, 3-, 6-, 9-, and 12-BP phenols and the BP 1,6-, 3,6-, 6,12-, and 9,10-quinones are now reported. The syntheses of the BP phenols (except 6-HO-BP) involve the key steps of Pd-catalyzed Suzuki Miyaura cross-coupling of a naphthalene boronate ester with a substituted aryl bromide or triflate ester. The BP quinones were synthesized from the corresponding BP phenols by direct oxidation with the hypervalent iodine reagents IBX or TBI. These reagents exhibited different regiospecificities. IBX oxidation of the 7- and 9-BP phenols provided the ortho-quinone isomers (BP 7,8- and 9,10 diones, respectively), whereas TBI oxidation of the 1-, 3-, and 12-BP phenols furnished BP quinone isomers with carbonyl functions in separate rings (BP 1,6-, 3,6-, and 6,12-diones, respectively). PMID- 19132943 TI - Marinisporolides, polyene-polyol macrolides from a marine actinomycete of the new genus Marinispora. AB - Two new polyene macrolides, marinisporolides A and B (1, 2), were isolated from the saline culture of the marine actinomycete, strain CNQ-140, identified as a member of the new marine genus Marinispora. The marinisporolides are 34-membered macrolides composed of a conjugated pentaene and several pairs of 1,3-dihydroxyl functionalities. Marinisporolide A (1) contains a bicyclic spiro-bis tetrahydropyran ketal functionality, while marinisporolide B (2) is the corresponding hemiketal. The structures of these new compounds were assigned by combined spectral and chemical methods including extensive 2D NMR experiments and correlations of (13)C NMR data with Kishi's Universal NMR Database. Chemical modifications, including methanolysis, acetonide formation, and application of the modified Mosher method, provided the full stereostructures of these molecules. Three additional macrolides, marinisporolides C-E (3-5), which are olefin geometric isomers of marinisporolide A (1), were also isolated and their structures defined. Under room light, marinisporolides A and B readily photoisomerize to C-E indicating that they are most likely produced by photochemical conversion during the cultivation or isolation procedures. Although polyenes, marinisporolides A (1) and B (2) showed weak to no antifungal activity against Candida albicans. PMID- 19132944 TI - Divergent synthesis of three classes of aryl N-glycosides by solvent control. AB - Aryl glycosides represent a group of molecules with immense biological applications and implications. While the syntheses of aryl C-glycosides and O glycosides have been studied extensively, the preparation for aryl N-glycosides is relatively unexplored. By employing 1,4-naphthoquinone and glycosyl azides undergoing a [3 + 2] cycloaddition, we have developed a convenient method for constructing three different classes of aryl N-glycosides that include N glycosylated 2-aminomethylene-1,3-indanedione, benzazepine-1,5-dione, and 9,10 anthraquinone derivatives via solvent control. It was found that conducting cycloaddition in DMF formed exclusively 9,10-anthraquinone derivatives, while less polar solvent such as toluene offered all three aryl N-glycosides. The synthesis of N-glycosylated 9,10-anthraquinone derivatives is of particular interest since no known example has been documented. The synthesis of these N glycosylated heterocyclic compounds using traditional glycosylation methods could be challenging. Therefore, our diversity-oriented protocols can be viewed as an alternative and practical glycosylation approach. In addition, we have also demonstrated that alkyl azides can also undergo the same cycloaddition, further expanding the structural repertoire available for a broader interest. Initial anticancer assays have revealed that 19f and 19k exert mean growth percent of 17.58 and -5.95, respectively. PMID- 19132945 TI - Enantioselective nitroaldol reaction catalyzed by sterically modified salen chromium complexes. AB - A group of modified (salen)Cr(III)Cl complexes with bulky benzylic substituents in the 3,3'-position of the salicylidene moiety have been successfully applied for the asymmetric nitroaldol reaction. The readily accessible complex bearing 3 phenylpent-3-yl groups (2 mol %) leads to beta-nitro alcohols in up to 92% yield and 94% ee. PMID- 19132946 TI - Stereocontrolled synthesis of D- and L-beta-rhamnopyranosides with 4-O-6-S-alpha cyanobenzylidene-protected 6-thiorhamnopyranosyl thioglycosides. AB - The synthesis of both enantiomers of a 4-O-6-S-alpha-cyanobenzylidene-protected 6 thiorhamnopyranosyl thioglycoside is described starting from D-mannnose and L arabinose derivatives for the D- and L-series, respectively. This donor is effective in the preparation of the corresponding beta-glycosides using the 1 benzenesulfinyl piperidine/trifluoromethanesulfonic anhydride protocol. Following desulfurization and concomitant debenzylation with Raney nickel, the so-formed 6 thio-beta-mannosides are converted in high yield to the beta-rhamnopyranosides. PMID- 19132947 TI - A catalytic method for the preparation of polysubstituted cyclopentanes: [3+2] cycloaddition of vinylidenecyclopropanes with activated olefins catalyzed by triflic imide. AB - [3+2] Cycloadditions of vinylidenecyclopropanes (VDCPs) with electron-deficient olefins, such as methyl vinyl ketone (MVK) and acrylaldehyde, proceed smoothly in the presence of a catalytic amount of triflic imide (Tf(2)NH) to give the corresponding functionalized cyclopentanes in good to high yields. PMID- 19132949 TI - Unilateral myelinated nerve fibers associated with hypertropia, strabismus and amblyopia? Reverse straatsma syndrome? PMID- 19132950 TI - Overaction of the inferior oblique muscle in 4th nerve palsy. PMID- 19132951 TI - Whither scientific publication in the internet age: are books and hard copies going to disappear? Major aniseikonia review; amblyopia and the retina; binocular motility and alignment: the exoshift under general anesthesia revisited; Straatsma syndrome. Followups on civility and "system failure". PMID- 19132952 TI - Clinical usefulness of the Aniseikonia Inspector: a review. AB - PURPOSE: In the literature several articles have appeared which describe the accuracy and repeatability of direct-comparison aniseikonia tests, such as the New Aniseikonia Test and the aniseikonia test of the Aniseikonia Inspector. In these articles a discussion on the clinical relevance of the results is often missing. Therefore, this article discusses the clinical usefulness of direct comparison aniseikonia tests, in particular that of the Aniseikonia Inspector. METHOD: Review of the literature shows that an aniseikonia test is usually evaluated by inducing aniseikonia in normal subjects using size lenses. The range of induced aniseikonia differs. Some investigations evaluated from appr. -8% to +8%, while other investigations evaluate a (too) small range from appr. -3% to +3%. RESULTS: In general direct-comparison aniseikonia tests have a repeated measurement accuracy of approximately 0.5% standard deviation (this also includes actual aniseikonia fluctuations). A direct-comparison aniseikonia test with free eye rotation seems to underestimate the amount of aniseikonia. However, converting these underestimations into clinically applicable values shows that vertically the 'error' is on average smaller than only 0.5% and horizontally the 'error' is smaller than 1%. This may be statistically significant but clinically less relevant underestimation seems to disappear in tests without free eye rotation (as in the newer versions of the Aniseikonia Inspector). CONCLUSION: Because aniseikonia does not seem to give any clinical symptoms until the aniseikonia has a value of 3-5%, the accuracy and repeatability of direction comparison tests, such as that of the Aniseikonia Inspector, is sufficient for effective aniseikonia management. PMID- 19132953 TI - The exoshift under anesthesia correlates with probable changes in medial rectus innervation after surgery for infantile esotropia. AB - PURPOSE: To study the outward deviation of the eyes (exoshift) under anesthesia, in a variety of clinical settings in order to improve our understanding of how medial rectus recessions change alignment and innervation. METHODS: Pre-operative and intraoperative eye deviations were measured before surgery and under Stage 3 of general anesthesia using a modified Krimsky test in 5 groups: 1) Unoperated infantile esotropia (N=60); 2) Undercorrected infantile esotropia (N=27); 3) Corrected infantile esotropia with subsequent vertical deviations (N=17); 4) Superior oblique palsies without horizontal deviations (N=21); and 5) Late consecutive exotropia (N=16). RESULTS: Group 2 averaged half the esotropic deviation of Group 1 (19.8 +/-7.4 ET vs. 42.1 +/-18.3 ET), but had a nearly identical exoshift (41.9 +/-13.2 vs. 41.8 +/-13.6, p=0.96). Group 4 (orthotropic) and Group 5 (exotropic) demonstrated smaller and nearly identical exoshifts (26.0 +/-8.3 vs.24.0 +/-9.3, p=0.50). Group 3 had significantly less exotropia (30.1 +/ 6.0) than Group 1 and 2 (p=0.002 for both), but significantly more exoshift than Group 4 (p=0.04) or Group 5 (p=0.067). CONCLUSION: Contracture of the lateral rectus reduced the deviations after undercorrecting surgery, but the exoshift remained unchanged. Medial rectus recession by itself has no effect on normal level seen in consecutive exotropia. These data combined with a reasonable set of assumptions regarding the state of contracture (expansure) of the horizontal rectus in a variety of pre- and post-operative settings lead to the conclusion that setting the eyes straight with a successful bilateral medical rectus recession reduces the pre-operative hyperinnervation of medial rectus almost to normal, with a small amount of residual hyperinnervation remaining to overcome the increased contacture of the lateral rectus which occurs due to increased exotropia during sleep after a bilateral medial rectus recession. When this small residual hyperinnervation decreases to normal, consecutive exotropia develops, owing to continued increased contracture of the lateral recti. PMID- 19132954 TI - Macular and retinal nerve fibre layer thickness in strabismus and anisometropic amblyopia. AB - Electrophysiological studies investigating the relationship between amblyopia and retinal function have in the past produced conflicting findings, leaving the retinal correlates of amblyopia, if they exist, unknown. Recent advances in technology, and in particular the emergence of optical coherence tomography, has resulted in further investigations of the retinal nerve fibre layer and macula of amblyopia patients using these instruments. This review summarises the recent literature in this area and discusses various issues relating to this research. PMID- 19132955 TI - The ABC transporter MsbA interacts with lipid A and amphipathic drugs at different sites. AB - MsbA is an essential ABC (ATP-binding cassette) transporter involved in lipid A transport across the cytoplasmic membrane of Gram-negative bacteria. The protein has also been linked to efflux of amphipathic drugs. Purified wild-type MsbA was labelled stoichiometrically with the fluorescent probe MIANS [2-(4' maleimidylanilino)naphthalene-6-sulfonic acid] on C315, which is located within the intracellular domain connecting transmembrane helix 6 and the nucleotide binding domain. MsbA-MIANS displayed high ATPase activity, and its folding and stability were unchanged. The initial rate of MsbA labelling by MIANS was reduced in the presence of amphipathic drugs, suggesting that binding of these compounds alters the protein conformation. The fluorescence of MsbA-MIANS was saturably quenched by nucleotides, lipid A and various drugs, and estimates of the Kd values for binding fell in the range of 0.35-10 microM. Lipid A and daunorubicin were able to bind to MsbA-MIANS simultaneously, implying that they occupy different binding sites. The effects of nucleotide and lipid A/daunorubicin binding were additive, and binding was not ordered. The Kd of MsbA for binding lipid A was substantially decreased when the daunorubicin binding site was occupied first, and prior binding of nucleotide also modulated lipid A binding affinity. These results indicate that MsbA contains two substrate-binding sites that communicate with both the nucleotide-binding domain and with each other. One is a high affinity binding site for the physiological substrate, lipid A, and the other site interacts with drugs with comparable affinity. Thus MsbA may function as both a lipid flippase and a multidrug transporter. PMID- 19132956 TI - A common variant of the eNOS gene (E298D) is an independent risk factor for left ventricular hypertrophy in human essential hypertension. AB - eNOS (endothelial NO synthase) plays a critical role in the development of ventricular remodelling and cardiac hypertrophy. The purpose of the present study was to determine whether three common variants in NOS3 (the eNOS gene) are associated with the risk of LVH [LV (left ventricular) hypertrophy] in patients with essential hypertension. Three NOS3 genetic variants, -T786C (rs2070744), eNOS4a/b and +G894T (rs1799983), were genotyped in two independent case-control studies: the first study consisted of 1061 hypertensive patients with LVH and 1118 hypertensive patients without LVH, and the second sample consisted of 120 patients with LVH and 223 patients without LVH. Echocardiographic measurements were obtained in all of the hypertensive patients. Only the +G894T (E298D) variant of NOS3 was associated with a higher risk of LVH {OR (odds ratio), 1.67 [95% CI (confidence interval), 1.19-2.36]; P<0.01} in the first population, and replicated in the second population [OR, 1.41 (95% CI, 1.01-2.28); P<0.05] in a recessive model. Compared with carriers of the G allele (GT+GG), patients carrying the TT genotype had increased septal wall thickness (16.2%, P<0.01 and 11.7%, P<0.01 respectively), LV posterior wall thickness (8.3%, P<0.01 and 7.1%, P<0.01 respectively), LV mass index (14.0%, P<0.01 and 25.1%, P<0.01 respectively) and relative wall thickness (13.1%, P<0.01 and 16.2%, P<0.01 respectively) in the first and second populations. The results of the present study support that homozygosity for +G894T (E298D) in NOS3 is a genetic risk factor for the development of LVH in patients with hypertension. PMID- 19132957 TI - Do resistin-induced changes in tissue blood perfusion play a role in the pathogenesis of type 2 diabetes? PMID- 19132958 TI - Insulin as ischaemic preconditioning-mimetic. PMID- 19132960 TI - Preface: treatment optimization in schizophrenia through active patient management - proceedings from two European consensus meetings. PMID- 19132961 TI - Definitions of response and remission in schizophrenia: recommendations for their use and their presentation. AB - OBJECTIVE: To review and make recommendations for the definition and presentation of the terms 'response' and 'remission' in schizophrenia. METHOD: Selective review of publications on definitions of response and remission in schizophrenia. RESULTS: When the Brief Psychiatric Rating Scale (BPRS) or the Positive and Negative Syndrome Scale (PANSS) are used for definitions of response, a cut-off of at least 50% reduction of the baseline score should be used for acutely ill, non-refractory patients and a cut-off of at least 25% reduction for refractory patients. When percentage BPRS/PANSS reduction is calculated, the 18/30 points minimum scores meaning 'no symptoms' on the should be subtracted. In addition, responder rates from 0-100% could be presented in a table in steps of 25%. For large and simple practical trials, the Clinical Global Impression scale with suggested improvements could be used 1-7 scale. CONCLUSION: To show how many patients are still symptomatic at the end of study and to show the overall amount of change in both remission and responder criteria should be presented. PMID- 19132962 TI - Schizophrenia and physical health problems. AB - OBJECTIVE: To estimate the prevalence of physical health problems in patients with schizophrenia, and to appraise the impact on mortality rates and quality of life (QoL) in such patients. METHOD: A selective review of clinical articles relating to physical health such as cardiovascular disease, metabolic syndrome and QoL. In addition, current guidelines and recommendations for the monitoring of physical health in schizophrenia were reviewed. RESULTS: Cardiovascular events contribute most strongly to the excess mortality observed in schizophrenia. Other factors that contribute significantly include obesity, metabolic aberrations, smoking, alcohol, lack of exercise and poor diet - all of which might be targets for health promoting activities. CONCLUSION: Physical health problems in patients with schizophrenia are common, and contribute to the excess mortality rate, as well as decreasing QoL. Many adverse physical factors are malleable in such patients, and physical benefit may be gained by following practical guidelines for their monitoring and improvement. PMID- 19132963 TI - Side-effects of antipsychotic medication and health-related quality of life in schizophrenia. AB - OBJECTIVE: This analysis used data from the large (n = 1208) European Schizophrenia Cohort to examine the association between subjective side-effects of antipsychotic medication and the Mental and Physical Composite Scores (MCS; PCS) of the SF-36 scale. METHOD: Relationships between the subjective evaluation of side-effects identified from the Subjective Side-Effects Scale and the adjusted mean score on the PCS and MCS were examined. Where appropriate, these associations of subjective side-effects were compared with those of the same side effects measured objectively. RESULTS: In this study, subjective side-effects of antipsychotic medication were linked either to both the PCS and the MCS or, in a few instances, to neither. Subjective evaluations of sexual side-effects were associated only with the MCS, those of sialorrhoea only with the PCS. Objective ratings of extrapyramidal side-effects were related neither to PCS nor to MCS. CONCLUSION: These data suggest that side-effects, whether subjective or objective, may need to be considered individually in relation to their impact on quality of life. PMID- 19132964 TI - Schizophrenia treatment: content versus delivery. AB - OBJECTIVE: To review the evidence supporting the importance of ensuring that patients with psychiatric disorders receive an optimal and appropriate level of non-pharmacological treatment, and how Assertive Community Treatment (ACT) may be able to contribute to this aim. METHOD: Analysis of data from selected individual published studies on ACT, in addition to reviews from the Cochrane Library, and other study groups. RESULTS: Treatment management using ACT appears to offer benefits in terms of reduction in hospitalisation, although there is some debate as to whether this is the most representative outcome measure. Preliminary indications using remission as an outcome measure have also shown promising results in favour of ACT. CONCLUSION: While further investigation and validation are necessary, current data indicate that ACT may be an appropriate strategy to facilitate the delivery of treatment to patients with psychotic disorders. PMID- 19132965 TI - Connecting patient needs with treatment management. AB - OBJECTIVE: To propose ideas for the development of a core strategy for monitoring patients with schizophrenia to ensure physical health and optimal treatment provision. METHOD: A panel of European experts in the field of schizophrenia met in Bordeaux in June 2006 to discuss, 'Patient management optimisation through improved treatment monitoring.' RESULTS: Key consensus from the discussion deemed that weight gain, oral health and ECG parameters were core baseline parameters to be monitored in all patients with schizophrenia. Further, an identification of a patient's own barriers to treatment alongside local health service strategies might comprise elements of an individualised management strategy which would contribute to optimisation of treatment. Any monitoring strategy should be kept simple to encourage physician compliance. CONCLUSION: A practical solution to the difficulties of providing holistic patient care would be to suggest a limited set of physical parameters to be monitored by physicians on a regular basis. PMID- 19132966 TI - Treatment optimisation in schizophrenia through active patient management, proceedings from two European consensus meetings. Epilogue. PMID- 19132968 TI - More statistical analysis is needed for the meta-analysis of genetic association studies. PMID- 19132972 TI - Rhinitis and asthma represent hot topics for allergy. PMID- 19132970 TI - The safety profile of anti-tumour necrosis factor therapy in inflammatory bowel disease in clinical practice: analysis of 620 patient-years follow-up. AB - BACKGROUND: Anti-TNF agents are now widely used in Crohn's disease (CD), and in ulcerative colitis (UC). AIM: To review the safety profile of anti-TNF agents in all patients treated with infliximab in Edinburgh from 1999 to 2007. METHODS: Complete data were available on 202/207 patients comprising 157 CD, 42 UC and three coeliac disease. Median follow-up was 2.4 years (1.0-4.9) with a total of 620 patient-years follow-up. About 19.1% of CD patients were subsequently treated with adalimumab. RESULTS: Seven deaths (3.3%) occurred in follow-up; only one death was <1 year post-infliximab (at day 72, from lung cancer). A total of six malignancies (three haematological, three bronchogenic) and six cases of suspected demyelination (three with confirmed neurological disease) were reported. In the 90 days following infliximab, 95 adverse events (36 serious) occurred in 58/202 (28.7%) patients. In all, 42/202 (20.8%) had an infectious event (22 serious) and 27/202 (13.4%) of patients had an infusion reaction: 19 acute (four serious) and eight delayed (three serious). CONCLUSIONS: Serious infections, malignancies and neurological disease complicate anti-TNF use in clinical practice. Although evidence for causality is unclear, potential mechanisms and predisposing factors need to be explored. In individual patients, the risk/benefit analysis needs to be carefully assessed and discussed prior to commencement of therapy. PMID- 19132973 TI - Dysregulation of the stress response in asthmatic children. AB - The stress system co-ordinates the adaptive responses of the organism to stressors of any kind. Inappropriate responsiveness may account for increased susceptibility to a variety of disorders, including asthma. Accumulated evidence from animal models suggests that exogenously applied stress enhances airway reactivity and increases allergen-induced airway inflammation. This is in agreement with the clinical observation that stressful life events increase the risk of a new asthma attack. Activation of the hypothalamic-pituitary-adrenal (HPA) axis by specific cytokines increases the release of cortisol, which in turn feeds back and suppresses the immune reaction. Data from animal models suggest that inability to increase glucocorticoid production in response to stress is associated with increased airway inflammation with mechanical dysfunction of the lungs. Recently, a growing body of evidence shows that asthmatic subjects who are not treated with inhaled corticosteroids (ICS) are likely to have an attenuated activity and/or responsiveness of their HPA axis. In line with this concept, most asthmatic children demonstrate improved HPA axis responsiveness on conventional doses of ICS, as their airway inflammation subsides. Few patients may experience further deterioration of adrenal function, a phenomenon which may be genetically determined. PMID- 19132974 TI - Topical glucocorticoids downregulate COX-1 positive cells in nasal polyps. AB - BACKGROUND: Influx of inflammatory cells is one of the hallmarks of nasal polyposis. As glucocorticoids (GC) are known to exhibit strong anti-inflammatory effects, these drugs are frequently used in the treatment of the disease. Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis. As cyclooxygenases (COX) are key enzymes in the synthesis of both pro- (COX-1, COX-2) and anti-inflammatory prostanoids (COX-2), we investigated the role of topical GC on COX-1, COX-2 and inflammatory markers in nasal polyps (NP). METHODS: Immunohistochemical analysis of inflammatory markers (CD68, CD117, MBP, elastase, IgE, BB-1, IL-4, IL-5 and IL-6), COX-1 and COX-2 was performed on normal nasal mucosa (NM) (n = 18), non-GC treated NP (n = 27) and topical GC treated NP (n = 12). NP groups were matched for allergy, asthma and ASA intolerance. RESULTS: Increased numbers of eosinophils, IL-5+ cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation (P < 0.05). In addition, increased numbers of COX-1+ cells are observed in NP epithelium compared to NM (P < 0.05). CONCLUSION: Topical GC significantly reduce the number of COX-1+ NP cells (P < 0.05), but have no significant effect on COX 2+ NP cells. No significant reduction in the number of eosinophils is observed for GC treated NP. The number of IL-5+ cells is however increased significantly upon GC treatment (P < 0.05). PMID- 19132975 TI - Global map of the prevalence of symptoms of rhinoconjunctivitis in children: The International Study of Asthma and Allergies in Childhood (ISAAC) Phase Three. AB - BACKGROUND: Phase One of the International Study of Asthma and Allergies in Childhood (ISAAC) measured the global patterns of prevalence and severity of symptoms of rhinoconjunctivitis in children in 1993-1997. METHODS: International Study of Asthma and Allergies in Childhood Phase Three was a cross-sectional survey performed 5-10 years after Phase One using the same methodology. Phase Three covered all of the major regions of the world and involved 1 059 053 children of 2 age groups from 236 centres in 98 countries. RESULTS: The average overall prevalence of current rhinoconjunctivitis symptoms was 14.6% for the 13- to 14-year old children (range 1.0-45%). Variation in the prevalence of severe rhinoconjunctivitis symptoms was observed between centres (range 0.0-5.1%) and regions (range 0.4% in western Europe to 2.3% in Africa), with the highest prevalence being observed mainly in the centres from middle and low income countries, particularly in Africa and Latin America. Co-morbidity with asthma and eczema varied from 1.6% in the Indian sub-continent to 4.7% in North America. For 6- to 7-year old children, the average prevalence of rhinoconjunctivitis symptoms was 8.5%, and large variations in symptom prevalence were also observed between regions, countries and centres. DISCUSSION: Wide global variations exist in the prevalence of current rhinoconjunctivitis symptoms, being higher in high vs low income countries, but the prevalence of severe symptoms was greater in less affluent countries. Co-morbidity with asthma is high particularly in Africa, North America and Oceania. This global map of symptom prevalence is of clinical importance for health professionals. PMID- 19132978 TI - Identifying rarer genetic variants for common complex diseases: diseased versus neutral discovery panels. AB - The power of genetic association studies to identify disease susceptibility alleles fundamentally relies on the variants studied. The standard approach is to determine a set of tagging-SNPs (tSNPs) that capture the majority of genomic variation in regions of interest by exploiting local correlation structures. Typically, tSNPs are selected from neutral discovery panels - collections of individuals comprehensively genotyped across a region. We investigated the implications of discovery panel design on tSNP performance in association studies using realistically-simulated sequence data. We found that discovery panels of 24 sequenced 'neutral' individuals (similar to NIEHS or HapMap ENCODE data) were sufficient to select well-powered tSNPs to identify common susceptibility alleles. For less common alleles (0.01-0.05 frequency) we found neutral panels of this size inadequate, particularly if low-frequency variants were removed prior to tSNP selection; superior tSNPs were found using panels of diseased individuals. Only large neutral panels (200 individuals) matched diseased panel performance in selecting well-powered tSNPs to detect both common and rarer alleles. The 1000 Genomes Project initiative may provide larger neutral panels necessary to identify rarer susceptibility alleles in association studies. In the interim, our results suggest investigators can boost power to detect such alleles by sequencing diseased individuals for tSNP selection. PMID- 19132979 TI - Retraction. Evaluation of the NK2 homeobox 1 gene (NKX2-1)as a Hirschsprung's disease locus. Ann Hum Genet 72, 170-177. PMID- 19132976 TI - Comparison of the efficacy and safety of bilastine 20 mg vs desloratadine 5 mg in seasonal allergic rhinitis patients. AB - BACKGROUND: Bilastine is a novel, nonsedating H(1)-antihistamine developed for symptomatic treatment of Allergic Rhinitis and Chronic Idiopathic Urticaria. The objective of this study was to compare the efficacy and safety of bilastine 20 mg vs placebo and desloratadine 5 mg in subjects with seasonal allergic rhinitis (SAR). METHODS: This randomized, double blind, placebo-controlled, parallel-group multicentre study evaluated the effect of 2 weeks' treatment with bilastine 20 mg, desloratadine 5 mg or matched placebo once daily, in 12-70 years old symptomatic SAR patients. All subjects assessed the severity of nasal (obstruction, rhinorrhoea, itching, and sneezing) and nonnasal (ocular itching, tearing, ocular redness, itching of ears and/or palate) symptoms on a predetermined scale to provide a total symptom score (TSS), composed of nasal and nonnasal symptom scores (NSS and NNSS, respectively). The primary efficacy measure was the area under the curve (AUC) for the TSS over the entire treatment period. RESULTS: Bilastine 20 mg significantly reduced the AUC of TSS to a greater degree from baseline compared to placebo (98.4 with bilastine vs 118.4 with placebo; P < 0.001), but not compared to desloratadine 5 mg (100.5). Bilastine 20 mg was not different from desloratadine 5 mg but significantly more effective than placebo in improving the NSS, NNSS, and rhinitis-associated discomfort scores (P < 0.05), and rhinoconjunctivitis quality of life questionnaire total (P < 0.005) and four out of seven individual domain (P < 0.05) scores. The incidence of treatment emergent adverse events was similar for bilastine (20.6%), desloratadine (19.8%), and placebo (18.8%). CONCLUSION: Bilastine 20 mg once daily was efficacious, safe and not different from desloratadine 5 mg once daily in the treatment of SAR symptoms. PMID- 19132980 TI - A novel monoclonal antibody recognizing beta(1-3) glucans in intact cells of Candida and Cryptococcus. AB - The cell walls of all medically important fungi contain a unique polyglucose compound, beta(1-3) glucan. In the present study, murine monoclonal antibodies were produced against linear and beta(1-6) branched beta(1-3) glucans, and their specificities were characterized for reactivity to other beta glucans, fungal cell wall fragments, and fungal cells. Their reactivity was also compared with that of rabbit polyclonal antibodies raised against the same immunogens. Two mouse monoclonal antibodies (AG and BG) recognized immunoreactive epitopes in beta(1-3)(1-6) glucan by ELISA. In an inhibition assay of the anti-beta(1-3)(1-6) activity of the monoclonals, the homologous antigen effectively inhibited the activity as expected, while beta(1-3) also inhibited the assay but to a much lesser extent. No inhibition was obtained by beta(1-3)(1-4) or beta(1-6), while a cell wall extract of Candida albicans (PPM) effectively inhibited both monoclonals. Cell wall fragments of C. albicans (CaCW) and Cryptococcus neoformans (CnCW) inhibited the anti-beta(1-3)(1-6) activity of AG, while BG was much less or not inhibited at all. Immunofluorescence confirmed the unique antibody specificity of AG by its recognition of a beta(1-3)(1-6)-associated epitope on the cell surfaces of C. albicans,C. krusei, C. glabrata, and nonencapsulated C. neoformans. The epitope for the AG antibody is suggested to be present in the branching point of beta(1-3)(1-6), or in the randomly coiled beta(1-3) polyglucan due to the presence of branches. Thus, monoclonal antibodies to beta(1-3)(1-6) glucans may have potential as tools in the laboratory diagnosis of invasive yeast infections. PMID- 19132982 TI - Heat shock proteins 27, 60 and 70 as prognostic markers of prostate cancer. AB - Heat shock proteins (HSPs) protect cells against stress-associated injury and are overexpressed in several malignant tumors. We aimed to investigate their value as prognostic markers in prostate cancer. A tissue microarray (TMA) was constructed of 289 prostate cancers from radical prostatectomy (RP) specimens with median follow-up of 48.9 months. Slides were immunostained for HSP27, HSP60 and HSP70. Intensity and extent of immunoreactivity (IR) and their product (IRp) was evaluated by two observers. The IRp of HSP27 and HSP60, but not of HSP70, significantly predicted biochemical recurrence (p=0.014, 0.034 and 0.160, respectively). Recurrence-free survival in patients with strong HSP27 and HSP60 staining was shorter than in those with weak expression (p=0.019 and 0.001, respectively). IRp of HSP27 and HSP60 correlated with Gleason score (p<0.01). HSP60 was an independent predictor of biochemical recurrence in multivariate analysis, including extraprostatic extension, margin status, seminal vesicle invasion and Gleason score. Weighted kappa for interobserver agreement of HSP27, HSP60 and HSP70 IR was 0.613-0.823 for intensity and 0.584-0.719 for IRp, but only 0.036-0.244 for extent, raising the question whether staining extent should be estimated on TMA. We conclude that HSP27 and HSP60 are predictors of biochemical recurrence after RP. PMID- 19132981 TI - Clinical isolates of Staphylococcus aureus from the Arkhangelsk region, Russia: antimicrobial susceptibility, molecular epidemiology, and distribution of Panton Valentine leukocidin genes. AB - A total of 91 consecutive clinical isolates of Staphylococcus aureus were collected at the Regional Hospital of Arkhangelsk, Russia, from May to December 2004, and examined for antimicrobial susceptibility, methicillin resistance and presence of Panton-Valentine leucocidin (PVL) genes. Epidemiological typing was performed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Methicillin-resistant S. aureus (MRSA) isolates were examined by staphylococcal cassette chromosome mec (SCCmec) typing. High-to-moderate rates of resistance to penicillin (beta-lactamase production; 93%), tetracycline (40%), erythromycin and clindamycin (32%) were observed. Forty out of ninety-one (44%) isolates were positive for PVL genes. Thirty-six (40%) PVL-positive methicillin susceptible S. aureus (MSSA) strains were shown by PFGE and MLST typing (ST121, ST681, ST837) to be part of a nosocomial outbreak caused by clonal complex (CC) 121. PFGE, MLST and SCCmec typing revealed three MRSA clones. Sequence type (ST) 239-III (n=11), ST1097-III (n=1) and ST8-IV (n=3) belong to CC8 of epidemic multiresistant MRSA, whereas ST426-MRSA-IV/CC395 (n=1) has not been reported previously. All MRSA strains were PVL negative. The overall results underline the necessity of microbiological sampling, antimicrobial susceptibility testing, and epidemiological typing as a rational basis for antimicrobial treatment of S. aureus infections, and infection control measures to limit the spread of multiresistant MRSA and epidemic MSSA clones. PMID- 19132984 TI - Stereological quantification of tumor volume, mean nuclear volume and total number of melanoma cells correlated with morbidity and mortality. AB - Stereological quantification of tumor volume, total number of tumor cells and mean nuclear volume provides unbiased data, regardless of the three-dimensional shape of the melanocytic lesion. The aim of the present study was to investigate whether these variables are reproducible and may represent potential indicators of prognosis. Sixty patients who underwent surgery at the Department of Plastic Surgery, Aarhus University Hospital, from 1991 to 1994 were included in the study. Total tumor volume was estimated by the Cavalieri technique, total number of tumor cells by the optical dissector principle, and nuclear mean volume by point counting. Tumor volume, total number of tumor cells and nuclear mean volume were all reproducible parameters, showing a non-significant intra-observer variation (p=0.72; p=0.83, and p=0.15). Grouped according to the median values, the disease-free survival time was significantly different for tumor volume as well as total number of tumor cells (p=0.0006 and p=0.0004), but no significant difference in mortality was found (p=0.1 and p=0.06). The mean nuclear volume showed no difference regarding disease-free survival or mortality (p=0.5 and p=0.8). Breslow's thickness showed a significant impact on both disease-free survival (p=0.001) and mortality (p=0.009). In conclusion, tumor volume and total number of cancer cells were highly reproducible but did not add additional, independent prognostic information regarding the study population. PMID- 19132983 TI - Higher FoxP3 mRNA expression in peripheral blood mononuclear cells of GAD65 or IA 2 autoantibody-positive compared with autoantibody-negative persons. AB - The role of regulatory T cells (Tregs) in type 1 diabetes has been studied extensively. The most prevalent way to define Tregs has been by their surface expression of CD4 and CD25. As currently the transcription factor FoxP3 and the low expression of CD127 are regarded to be the most specific markers of Tregs, we analysed the number of Tregs defined by these molecules in peripheral blood mononuclear cells of diabetic patients and healthy controls. The gene expression of transforming growth factor beta and two isoforms of FoxP3 was measured as well. There were no significant differences between diabetic patients and healthy controls regarding the number of Tregs, or the expression of FoxP3 isoforms and TGFbeta in peripheral blood mononuclear cells. However, we found significantly higher expression of both full-length and Delta2FoxP3 in study subjects, positive for either GAD65 or IA-2 autoantibodies. The ratio of the expression of different isoforms was not changed. This study shows the possible role of FoxP3 in the development of tissue characteristic humoral immunity in type 1 diabetes. PMID- 19132985 TI - Prognostic value of COX-2 immunohistochemical expression evaluated by quantitative image analysis in colorectal cancer. AB - Epidemiological studies have shown that the inducible form of cyclooxygenase (COX 2) may be involved in colorectal carcinogenesis, but it is controversial whether its expression is a prognostic factor for colorectal cancer. The aim of the study was to examine the expression of COX-2 in colorectal cancer and investigate its prognostic relevance. Tissue sections of primary tumors from 132 patients undergoing curative resection for colorectal cancer were immunohistochemically examined for COX-2 expression. The levels of intensity and extent of COX-2 staining were quantified by use of a computerized image analysis system and correlated with various clinicopathological characteristics and survival. COX-2 immunoreactivity was observed in the cytoplasm of tumour epithelial cells of all colorectal cancer tissues examined. No significant correlation was found between levels of intensity and extent of COX-2 staining and various clinicopathological characteristics, including age, gender, tumor location, tumor size, tumor grade, depth of invasion, lymph node status and TNM stage. There was an inverse correlation between intensity and extent of COX-2 staining scores (Spearman's rho=-0.414; p<0.001). To analyze the prognostic value of intensity and extent of COX-2 staining, the patients were divided into four groups with respect to quartiles (< or =25; >25 to < or =50; >50 to < or =75; and >75). No significant disease-specific survival difference among the quartiles was found based on analysis of intensity (p=0.689) and extent (p=0.975) of COX-2 staining. These results suggest that the expression of COX-2 protein has no significant impact on the outcome of patients with colorectal cancer. PMID- 19132986 TI - Oestrogen receptor-beta1 but not oestrogen receptor-betacx is of prognostic value in apocrine carcinoma of the breast. AB - Apocrine carcinoma of the breast, which frequently expresses oestrogen receptor beta (ER-beta) in the absence of ER-alpha and only infrequently is treated endocrinologically, gives an opportunity to investigate the clinicopathological role of ER-beta in breast cancer independent of ER-alpha expression or tamoxifen treatment. Several isotypes of ER-beta, ER-beta1-5 etc., have been identified thus far; however, the clinicopathological importance of each ER-beta isotype in breast cancer is still uncertain. Here we aimed to clarify the clinicopathological importance of ER-beta1 and ER-betacx (ER-beta2) in apocrine carcinomas, immunohistochemically examining expressions of ER-beta1 and ER-betacx in 47 apocrine carcinomas. Positivity for ER-beta1 and ER-betacx was observed in 41 (87%) and 18 (38%) of 47 cases, respectively. ER-beta1 positivity was related to smaller tumor size (P=0.0359), lower histological grade (P=0.0322), and higher disease-free survival (P<0.0001), whereas ER-betacx status was related to none of these parameters. ER-beta1 positivity was also associated with favorable clinical outcome in 24 so-called triple-negative (ER-alpha-negative/PR-negative/HER2 negative) apocrine carcinomas. ER-beta1 itself, independent of ER-alpha expression and tamoxifen treatment, seems to have a tumor-suppressive effect, at least in apocrine carcinomas. Further study of ER-beta1 is desired to optimize breast cancer treatment. PMID- 19132987 TI - Absence of the AKT1 pleckstrin homology domain mutation in Japanese gastrointestinal and liver cancer patients. PMID- 19132988 TI - Renal oncocytosis and multiple papillary adenomas with oncocytoma as dominant nodule coexisting with papillary carcinoma in a patient with diabetic glomerulosclerosis, acquired renal cystic disease and B cell lymphoma. PMID- 19132989 TI - Somatic mutation of pro-cell death Bif-1 gene is rare in common human cancers. PMID- 19132990 TI - Undifferentiated carcinoma of the jejunum with extensive rhabdoid features. Case report and review of the literature. AB - Malignant rhabdoid tumor, first described in the kidney of young infants, is a rare and highly aggressive neoplasm of controversial histogenesis that has been reported at many other sites, including the gastrointestinal tract. However, malignant rhabdoid tumor of the small intestine is very rare, with only seven cases published to date. We report a 70-year-old man who presented with abdominal pain and weight loss, and showed a perforated jejunal mass with disseminated metastases by imaging. The patient underwent partial jejunectomy and biopsy of a liver metastasis. Microscopically, the tumor was characterized by neoplastic cells with vesicular nuclei, large nucleoli and abundant eccentric cytoplasm with hyaline globular intracytoplasmic inclusions. Immunohistochemically, the neoplasm coexpressed vimentin and epithelial antigens (AE1/AE3, Cam 5.2, CK34betaE12, CK19 and EMA), most of them showing a peculiar immunostaining pattern in relation to the globular inclusions. Ultrastructurally, the inclusions corresponded to paranuclear whorls of intermediate filaments. The patient received postoperative chemotherapy but died 9 months after surgery. In summary, we report the exceptional case of an undifferentiated carcinoma of the jejunum with rhabdoid phenotype. As with tumors at other sites, recognition of rhabdoid morphology in small intestine neoplasms is of significance because the prognosis is extremely poor. PMID- 19132991 TI - Serotonin content of normal and inflamed appendix: a possible role of serotonin in acute appendicitis. AB - The appendix is lined by a mucosa which has many neuroendocrine cells containing serotonin. Local release of serotonin can act as a mediator of inflammation. In this study we explored the serotonin content of the neuroendocrine cells of the appendixes removed for clinical diagnosis of appendicitis. Appendix specimens were divided into three groups: Acute appendicitis (AA), non-appendicitis (NA), and follicular hyperplasia (FH). Normal appendix specimens from patients undergoing elective abdominal surgery were used as the control group (NL). All sections were exposed to proteinase K, incubated with anti-serotonin, chromogranin A, and synaptophysin antibodies, and treated with the LSAB kit. Polygonal cells were seen within the crypt epithelium (enterochromaffin cell, EC) and within the lamina propria (subepithelial neuroendocrine cell, SNC). In AA, only 16 cases (64%) showed serotonin staining in non-destructed glands. There was a significant reduction in the number of ECs in AA compared to the FH (96%), NA (100%) and NL (100%) groups (P<0.001). Chromogranin and synaptophysin immunostaining also showed a significant reduction in the number of ECs in AA compared with the other three groups (P<0.001). SNC serotonin reactivity was lower in the AA group compared with the other groups (p<0.001). The inflamed appendix is markedly depleted of serotonin in the epithelium and lamina propria. Local serotonin release from ECs and SNCs in the appendix may act as an inflammatory mediator in appendicitis and is likely to be the source of raised blood serotonin in AA. PMID- 19132992 TI - Staphylococcus aureus as source of catheter-related bloodstream infection evaluated by PFGE and rep-PCR typing in a Brazilian hospital. AB - Staphylococci are a common cause of catheter-related bloodstream infection (CR BSI), and epidemiological typing is an important tool for effective infection control. This study evaluated by PFGE and rep-PCR whether Staphylococcus aureus strains isolated from skin and catheter tips were related to specimens isolated from blood. A prospective observational study, carried out in a clinical surgical ward at a Brazilian hospital between September 2000 and November 2002, investigated non-tunneled central venous catheters from 179 patients. S. aureus isolates were mainly obtained from blood (41.4%), while coagulase-negative staphylococci strains were more often isolated from the skin at the catheter insertion site (49.7%) and from the catheter tip (57.5%). Among the 21 strains isolated from 9 patients at 2 or 3 sites simultaneously, 9 were methicillin resistant S. aureus (MRSA) and 12 were methicillin-susceptible S. aureus (MSSA). Seven patients harbored the same S. aureus strain isolated from the skin, blood and/or catheter tip cultures. MRSA isolates belonged to one PFGE pattern (type A- subtypes A(1), A(2) and A(3)), and to two rep-PCR patterns (a and b). MSSA isolates were distinguished in five PFGE (B to F) and in three rep-PCR (c, d and e) patterns. Both PFGE and rep-PCR methods indicated that the skin at the catheter insertion site was the origin of CR-BSI caused by S. aureus. PMID- 19132993 TI - pS2 (TFF1) expression in prostate carcinoma: correlation with steroid receptor status. AB - pS2 or TFF1 is a member of the trefoil factor family, which is distributed throughout the gastrointestinal tract in both normal and diseased tissues. It is also considered to be one of the major estrogen-regulated proteins and an indicator of estrogen receptor (ER) functionality. pS2 has previously been investigated in benign and malignant prostate lesions with little information about its relationship to steroid receptor status. Our purpose was to correlate pS2 expression with steroid receptor status (ER alpha and progesterone receptor (PR)) and other pathologic variables in prostate carcinoma. 15 benign prostate hyperplasia (BPH) and 47 prostate carcinoma cases were investigated by means of immunohistochemistry for pS2, ER and PR expression. 80% of BPH showed pS2 cytoplasmic immunoreactivity in hyperplastic acini and about half of these cases also exhibited nuclear staining decorating basal or both basal and luminal nuclei. pS2 was highly expressed in prostate carcinoma (91.4%) with both cytoplasmic and nuclear patterns of staining. The latter pattern was significantly associated with carcinoma having a low Gleason score (p=0.02). pS2 lacked any significant correlation with steroid receptor status, stage or grade. Univariate survival analysis revealed a significant impact of stage (p=0.03) and nodal status (p<0.0001) on patient outcome. The diagnostic value of pS2 expression in prostate carcinoma validated 74.19% accuracy, 91.48% sensitivity and 78.18% positive predictive value. The high sensitivity of pS2 expression in prostate carcinoma could make it a suitable marker for diagnosis of prostate carcinoma, especially in metastatic cases of unknown origin. The absence of correlation and dissimilarity in immunolocalization between pS2 and ER alpha leads to the assumption that ER alpha could not be the regulatory protein for pS2 and may raise questions about the functionality of ER alpha in prostate. The nuclear pattern of pS2 immunoreactivity either in benign or malignant prostatic lesions is similar to the published data on ER beta distribution and could also identify a subset of carcinoma patients with a favorable prognosis. PMID- 19132994 TI - Pathological findings in rats with experimental allergic encephalomyelitis. AB - The aim of this study was to establish an animal model of experimental allergic encephalomyelitis (EAE) and examine the basic pathological changes, as well as expression and distribution of MMP-2 and MMP-9, in Wistar rats. Tissue sections were processed for HE staining, Weil myelin staining, and modified Bielschowsky staining. Expression and distribution of glial fibrillary acidic protein (GFAP), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) were detected with immunohistochemistry. We divided the EAE into five types, depending on pathological characteristics and clinical manifestations: acute EAE, relapsing remitting EAE, progressive EAE, benign EAE, and asymptomatic EAE. Rats with acute EAE suffered from quick, severe attacks with widespread inflammatory cells and axonal loss. No demyelination or astrocytic hyperplasia was found around the lesions. Rats with relapsing-remitting EAE broke down twice, with many perivascular cuffs and demyelinating plaques in lesions; hyperplastic and hypertrophic astrocytes characterized old lesions and axonal loss was evident. Rats suffering from progressive EAE exhibited continuous aggravation without improvement, accompanied by perivascular cuffs, demyelination, increased gliocytes and axonal damage. Rats with benign EAE recovered to a normal state with obviously decreased inflammatory cells and almost entirely unaffected myelin and axons. Rats with asymptomatic EAE also had various pathological changes that were not coincident with their clinical manifestations. Elevated expression of MMP-2 and MMP-9 was concordant in different types of EAE, but the extent differed in each type of EAE. MMP-2 and MMP-9 can be expressed in the form of vascular endothelial cells, meninges, or accumulated inflammatory cells. Multiple clinical courses of disease were demonstrated in Wistar rat EAE, with attributes similar to multiple sclerosis (MS) in clinical and pathological characteristics. Elevated expression of MMP-2 and MMP-9 may play a role in some aspects of pathological changes in EAE, for example, destroying the blood-brain barrier, degrading the myelin sheath, and damaging axons. PMID- 19132995 TI - Feeding of mice with Arabidopsis thaliana expressing the HIV-1 subtype C p24 antigen gives rise to systemic immune responses. AB - Development of transgenic edible plants, to be used as production, storage and delivery systems for recombinant vaccine antigens, is a promising strategy to obtain cost effective vaccines against infectious diseases, not least for use in developing countries. Therefore, we used Agrobacterium tumefaciens-mediated gene transfer to introduce the p24 gag gene encoding the nucleocapsid protein from HIV 1 subtype C into the Arabidopsis thaliana plant genome. Eighteen plant lines were confirmed positive for the p24 gene by PCR; four of these lines showed an apparent homozygous phenotype when grown on selective medium and these lines also showed transcription of the p24 gene into its corresponding mRNA. The mRNA in all four cases generated the p24 protein in plants, as verified by Western blot analysis. The plants were shown to contain between 0.2 mug and 0.5 mug p24 protein per g of fresh tissue. Analysis of the localisation of the p24 protein showed that stem tissue contained the largest amount of protein, more than twice as much as leaf tissue, whereas no p24 protein was detected in roots. By using Southern blotting, we found that 4, 2-3, 2 and 1 T-DNA insertion events took place in the four lines 1, 2, 7, and 10, respectively. The genetic insertions of line 1 were stable from the T(2) to the T(5) generation and gave rise to the p24 protein in all cases, as verified by Western blotting. In mice fed with fresh transgenic A. thaliana (line 10), anti-gag IgG was obtained in serum after a booster injection with recombinant p37Gag. No immune response was observed after equal booster injection of untreated mice or mice fed with A. thaliana WT plants. PMID- 19132996 TI - Expression of caspase-3-dependent apoptosis in mural thrombi leukocytes. AB - Failure of arteriovenous access is mostly due to graft thrombosis and multifactorial, with medical and surgical etiologies. Apoptosis of blood cells, such as macrophages, lymphocytes and eosinophils, may play an important role in thrombus formation. We also investigated caspase-3-dependent apoptosis in thrombi. We recorded clinical parameters in 43 consecutive patients with vascular access failure (13 men, 30 women; mean age+/-SD, 64.6+/-14.2 years) who underwent surgical thrombectomy. Major presentations included absent (92%) and/or near near absent (16%) flow through the access during hemodialysis. Cardiovascular risk factors were hypertension (70%), hyperlipidemia (47%), diabetes mellitus (47%), chronic obstructive pulmonary disease (12%), heart failure (12%), coronary artery disease (21%), and stroke (16%). Laboratory data included hemoglobin level of 100+/-17 g/L, total white blood cell count of 7.65+/-2.14 x 10(9)/L, and platelet count of 205.6+/-57.9 1000/iL. Abnormal biochemistry data included elevated blood urea nitrogen level of 63.5+/-24.4 mg/dL and creatinine level of 8.6+/-4.0 mg/dL (normal <1.4 mg/dL). Thrombi were characterized by apoptosis (32%) in a caspase dependent pathway in all types of leukocytes. Thrombi in arteriovenous access failure demonstrate apoptosis by means of the caspase-3 pathway in white blood cells. PMID- 19132997 TI - PEComata: highly melanotic multiple perivascular epithelioid cell tumors (PEComa) of the uterus. PMID- 19132998 TI - Real-time PCR and subsequent pyrosequencing for screening of penA mosaic alleles and prediction of reduced susceptibility to expanded-spectrum cephalosporins in Neisseria gonorrhoeae. PMID- 19132999 TI - Absence of ERCC6 gene mutation in common cancers of Korean patients. PMID- 19133000 TI - Black aortic valve--ochronosis. PMID- 19133001 TI - Carcinosarcoma arising in mixed epithelial and stromal tumor of the kidney. PMID- 19133002 TI - Multiple extragenital adenomatoid tumors in the mesocolon and omentum. AB - Adenomatoid tumors are benign mesothelial neoplasms most commonly found in the male and female genital tracts. Extragenital adenomatoid tumors are rare, most of them being solitary tumors. To our knowledge, only one case of multiple extragenital adenomatoid tumors, involving the liver and peritoneum, has been reported to date. Here we report another case of multiple extragenital adenomatoid tumors involving the mesocolon and omentum. A 47-year-old woman presented with a delayed menstrual period. Ultrasonography revealed a left adnexal mass, and surgical resection was attempted at a local hospital. The patient was transferred to our hospital without resection due to the intraoperative finding of multiple peritoneal tumors. At our hospital, an 8.0x7.5x6.0 cm tumor at the mesocolon of the sigmoid colon and three omental nodules measuring up to 2.5x2.0x1.7 cm were resected. Grossly, they were well circumscribed, gray-tan and elastic with small cystic spaces. Microscopically, they were composed of tubules and anastomosing channels lined by flattened or cuboidal cells with bland nuclei. Immunohistochemically, the tumor cells were positive for pan-cytokeratin AE1/AE3, vimentin, cytokeratin 5/6 and calretinin. The postoperative course was uneventful, and the patient was well 10 months after the operation. Despite their rarity, adenomatoid tumors should be included in the differential diagnosis of multiple intra-abdominal tumors. PMID- 19133003 TI - Solitary fibrous tumour of the kidney with sarcomatous overgrowth. Case report and review of the literature. AB - Solitary fibrous tumour (SFT) rarely occurs in the kidney, with only one case exhibiting malignant behaviour. We report the case of a typical SFT of the kidney with sarcomatous overgrowth in a 34-year-old woman. This malignant component, grossly apparent as a nodular area arising in the context of the main tumour mass, consisted of CD34+ mitotically active atypical plump spindle- to epithelioid-shaped cells, including pleomorphic multinucleated giant cells. A novel immunohistochemical finding was diffuse and strong S-100 protein expression by sarcomatous cells. This should be kept in mind by pathologists to avoid confusion with other S-100 protein-positive malignant neoplasms. PMID- 19133004 TI - Organ-confined clear cell renal cell carcinoma: the prognostic impact of microvascular invasion, nuclear grade and tumour size. AB - The frequency of diagnosed and treated organ-confined renal cell carcinoma is increasing. The prognosis of this group of tumours is difficult to predict. The main purpose of this study was to examine the prognostic significance of microvascular invasion, tumour size and nuclear grade in a complete cohort of 76 consecutive patients with organ-confined clear cell renal cell carcinoma treated with radical nephrectomy. Patient ages ranged from 39 to 88 years (mean 66 years). Median follow-up was 10.2 years (range 0.1-19.4 years). The tumours were graded according to Fuhrman. Representative histological sections were stained for CD31, which decorates endothelial cells, in order to assess microvascular invasion (MVI). In univariate analysis, microvascular invasion (p<0.01), tumour size (TS) (p=0.01), TNM stage (p=0.01) and Fuhrman nuclear grade (p=0.02) were significant predictors of cancer-specific survival. Multivariate analysis, adjusted for age, revealed that microvascular invasion, tumour size and nuclear grade were independent covariates. According to our findings microvascular invasion is a strong independent prognostic predictor, and including this in the histopathology report should be considered together with nuclear grade and tumour size. PMID- 19133005 TI - Expression of non-mast cell histidine decarboxylase in tumor-associated microvessels in human esophageal squamous cell carcinomas. AB - Histamine is produced by mast cells and many other types of cells. The role of histamine released from mast cells in promoting tumor angiogenesis has been intensively studied; however, the role of non-mast cell histamine in regulating tumor angiogenesis has been largely ignored. In this study, tissue specimen sections from 43 patients with esophageal squamous cell carcinoma (ESCC) and normal esophageal biopsies from 17 heath individuals obtained from a high incidence area of north China were used to assess changes in microvessel density (MVD) and non-mast cell L-histidine decarboxylase (HDC) (the only rate-limiting enzyme that catalyzes the formation of histamine from L-histidine) expression in the tumor microenvironment by immunohistochemistry (IHC). In addition, the cellular characterization of non-mast cell HDC-positive cells in microvessels was examined by double IHC combined with HDC/CD34 and HDC/PCNA antibodies. These IHC analyses revealed a significantly increased HDC-positive MVD in ESCC as compared with normal controls, which accounted for approximately 61% of CD34-labeled general MVD in ESCC. Furthermore, IHC in serial sections and double IHC showed that most of these HDC-positive cells were CD34-positive endothelial cells in microvessels with an increased proliferative capacity. Thus, our results suggest that non-mast cell histamine expressed in endothelial cells of microvessels could be an additional cellular source and might play a role in regulating angiogenesis in ESCC. PMID- 19133006 TI - Effect of interferon gamma and CD40 ligation on intracellular monocyte survival of nontypeable Haemophilus influenzae. AB - Nontypeable Haemophilus influenzae (NTHi) is a mucosal pathogen that is a major cause of respiratory infection, including sinusitis, otitis media and bronchitis. This bacterium has evolved a number of mechanisms to facilitate its survival in the human host. Recently it has been recognized that it is capable of intracellular survival in monocytes/macrophages and epithelial cells. Previous work by the authors has demonstrated that the protective response to NTHi is Th1 predominant. This information led to the hypothesis that the intracellular survival of NTHi in human monocytes may be reduced by two key effector mechanisms of Th1-mediated immunity: interferon gamma and ligation of CD40. This study assessed the effect of interferon gamma and ligation of CD40 on the intracellular survival of NTHi in human monocytes. Responses were studied in monocytes from subjects with bronchiectasis and persistent airway infection with NTHi and compared with control subjects. The results demonstrated that different isolates of NTHi were able to survive inside monocytes. Killing of one strain of NTHi could be enhanced by the addition of interferon gamma and CD40 ligation in both control and bronchiectasis subjects. Other strains were more resistant. PMID- 19133007 TI - Recurrence of pleomorphic adenoma of the parotid gland--predictive value of cadherin-11 and fascin. AB - The predictive value of cadherin-11, tenascin, fascin, and mucin-1 as markers for the likelihood of recurrence in pleomorphic adenoma of the parotid gland was examined. In this retrospective study we analysed 20 tumours from16 patients by immunohistochemistry. Staining intensities were measured using a semiquantitative scoring approach; localisation (tumour centre vs border) as well as clinical data were analysed and correlated with follow-up. Cadherin-11 was increased in recurrent tumours. However, no changes of fascin, tenascin or mucin-1 were observed. Cadherin-11 and fascin were increased in primary tumours of patients with later recurrence, with fascin upregulation restricted to the tumour border. In conclusion, cadherin-11 and fascin should be further analysed for their value as markers for later recurrence in pleomorphic adenoma. Our observations might reflect dysregulation of cellular pathways contributing to cellular dissemination, which might potentially result in later recurrence. PMID- 19133008 TI - Choice of endothelial marker is crucial for assessment of bone marrow microvessel density in chronic lymphocytic leukemia. AB - Angiogenesis is a potential prognostic factor in chronic lymphocytic leukemia (CLL). Elevated circulating levels of angiogenic factors in CLL have been repeatedly reported. Nevertheless, the issue of bone marrow neovascularization in CLL remains controversial, partly due to limited number of published studies, different methods of assessing microvessel density (MVD) and small patient cohorts. Moreover, there are very scarce data regarding the relationship of marrow angiogenesis to prognostic markers in CLL. Our objectives were: 1. To assess bone marrow MVD in CLL using two different monoclonal antibodies and a reproducible method of MVD quantification; 2. To examine the possible association of marrow MVD and clinical course, pattern of marrow infiltration, Rai stage, cytogenetic abnormalities detected by fluorescence in situ hybridization (FISH), and mutation status of immunoglobulin heavy chain variable region (IgVH). MVD was higher using CD34 vs vWF antibody (p<0.0001). However, no MVD differences were detected between CLL subgroups subdivided according to the above-mentioned prognostic factors. In conclusion, MVD assessment using anti-CD34 resulted in higher MVD counts than when using anti-vWF antibody. No association of MVD with any prognostic factors was observed, possibly due to the limited patient cohort. As the need for bone marrow trephine biopsies in CLL is significantly decreasing, a standardized method of neovascularization assessment is required to enable possible multicentre studies in order to conduct larger investigations and thereby shed more light on the real clinical significance of bone marrow angiogenesis in CLL. PMID- 19133009 TI - Intracellular replication of fusobacteria requires new actin filament formation of epithelial cells. AB - We examined survival and replication of fusobacteria inside epithelial cells. Subconfluent cultures of HaCaT keratinocytes were infected with five bacterial strains representing three Fusobacterium species: F. nucleatum, F. necrophorum, and F. mortiferum. Adhesion and invasion of the bacteria were assayed before and after antibiotic treatment that killed the adhered and extracellular bacteria. The number of live fusobacteria was examined by bacterial culturing after sonication of the epithelial cells. The role of host cell cytoskeleton functions was examined by treating the epithelial cells with cell function inhibitors. Number of viable epithelial cells was measured with the CellTiter96 kit. The tested Fusobacterium species adhered to and invaded the epithelial cells, and multiplied intracellularly for several hours. Thereafter, the intracellular number of bacteria rapidly declined. Concomitantly, viable fusobacteria were detected in the culture medium. Treatment of the infected epithelial cells with an actin formation inhibitor markedly reduced the number of living intracellular fusobacteria. Newly formed actin filaments were seen by confocal microscopy in the epithelial cells associated with the invaded bacteria. Fusobacteria infection did not reduce the number of viable epithelial cells in culture. Thus, fusobacteria are able to adhere to and invade epithelial cells, and survive under aerobic conditions. This property may enable them to survive in mucosa and participate in various disease processes of oral and pharyngeal tissues. PMID- 19133011 TI - Expression of chemokines and adhesion molecules in human coronary artery endothelial cells infected with Chlamydia (Chlamydophila) pneumoniae. AB - Chlamydia pneumoniae has during recent years been associated with cardiovascular disease and atherosclerosis. Chemokines, leukocyte adhesion proteins and metalloproteinases are significant for chemotaxis and attachment of leukocytes to vessel walls, and for stability of atherosclerotic plaques. To determine the ability of C. pneumoniae to elicit inflammation in a relevant target host cell, we infected human coronary artery endothelial cells (HCAEC) with a clinical isolate of C. pneumoniae. Extracellular release of five chemokines, two adhesion proteins and a metalloproteinase was measured at different time points after infection using a cytometric bead assay and ELISA. Secretion of IL-8, MCP-1, MIG, IP-10 and ICAM-1 was significantly increased 48 h after C. pneumoniae infection of HCAEC in comparison with uninfected controls. Release of RANTES occurred already 6 h after infection. C. pneumoniae did not elicit release of E-selectin or MMP-1. We conclude that C. pneumoniae induces expression of proinflammatory components in HCAEC, which would promote migration of leukocytes towards endothelial cells. This suggests that C. pneumoniae initiates and propagates vascular inflammation in ways that contribute to coronary artery disease. PMID- 19133010 TI - Immunogenicity and protective efficacy of a DNA vaccine encoding the fusion protein of mycobacterium heat shock protein 65 (Hsp65) with human interleukin-2 against Mycobacterium tuberculosis in BALB/c mice. AB - Developing a new generation of vaccines is important for preventing tuberculosis (TB). DNA vaccine is one promising candidate. In this study we evaluated the immunogenicity and protective efficacy of the DNA vaccine encoding the fusion protein of Mycobacterium tuberculosis heat shock protein 65 (Hsp65) with human interleukin-2 (hIL-2) in BALB/c mice. We showed that the DNA vaccine pcDNA-Hsp65 hIL-2 could induce high levels of antigen-specific antibody, IFN-gamma, CD4(+) and CD8(+) T cell production. When the immunized mice were infected with M. tuberculosis H37Rv, the organ bacterial loads in the DNA immunized group were significantly reduced compared to those of the saline control group, but the ability to reduce bacteria was not better than for BCG. The histopathology in lungs of the DNA vaccine immunized mice was similar to that of BCG immunized mice, which was obviously ameliorated compared to that of the saline control group. Overall, the DNA vaccine could afford protection against M. tuberculosis infection, though the protection efficacy was not as great as that of conventional BCG. PMID- 19133012 TI - Expression of L protein of Hantaan virus 84FLi strain and its application for recovery of minigenomes. AB - Hantaan virus (HTNV) is an Old World hantavirus associated with hemorrhagic fever with renal syndrome (HFRS). To visualize the localization of the L protein of HTNV strain 84FLi within cells, a fusion protein composed of enhanced green fluorescent protein and L protein, EGFP-L, was expressed in Vero cells. The 273 KDa expressed fusion protein of EGFP-L localized in the perinuclear region. We also described the development of a reverse genetics system for HTNV strain 84FLi. The RNA polymerase I (pol I)-mediated transcription system was used to generate artificial viral RNA genome segments (minigenomes), which contained the chloramphenicol acetyltransferase (CAT) reporter gene in antisense (virus RNA) or sense (virus-complementary RNA) orientation flanked by the noncoding regions of HTNV 84FLi L segment. CAT could be detected in cells after transfection, indicating the successful encapsidation, transcription and replication of the pol I-derived minigenomes. The passaged transfer of CAT demonstrates that recombinant virus containing packaged pol I-derived minigenomes has been produced. This system may be helpful in studying the gene function and pathogenesis of HTNV. PMID- 19133013 TI - Enhanced phosphorylation of the epidermal growth factor receptor at the site of tyrosine 992 in esophageal carcinomas. AB - This study aimed to determine whether epidermal growth factor receptor (EGFR), which has been reported to be frequently overexpressed in esophageal carcinoma cells, is actually activated in the cells. Paraffin-embedded specimens of 39 cases of esophageal carcinoma were analyzed immunohistochemically with anti-EGFR polyclonal antibody (alpha-EGFR Ab) and also an anti-phospho-EGFR-specific polyclonal antibody (alpha-p-EGFR(Tyr992) Ab) that specifically recognizes phosphorylated tyrosine 992 of EGFR. All of the 39 cases were found to express EGFR, but the expression levels were not significantly higher than those in basal cells of the normal esophageal epithelium. In 38 of the 39 cases, alpha-p EGFR(Tyr992) immunoreactivity was evident. Interestingly, the positively stained carcinoma cells were not distributed diffusely, and strongly immunostained cells tended to be localized in areas of severe dysplasia and in microinvasive foci just adjacent to the main invasive carcinoma. However, the deeply invasive front never exhibited positive immunoreactivity. The present findings suggest that phosphorylation of EGFR( Tyr992) may play some specific functional role in esophageal carcinomas besides promotion of cell proliferation. PMID- 19133014 TI - National case-control study on risk of death and Neisseria meningitidis sequence types in Denmark, 2000-2007. AB - In the past decades efforts to further diminish the case-fatality rate from meningococcal disease have proven challenging due to the often rapid progression of the disease in patients. In this study our objective was to characterise a subset of Neisseria meningitidis isolates to establish which sequence types were associated with increased mortality in Denmark during the period 2000-2007. We designed a matched case control and performed serogrouping, serotyping, serosubtyping and multilocus sequence typing (MLST) on 100 isolates. The clonal complex ST-32/ET-5 was found in 36% of the isolates, followed by the ST-11/ET-37 complex (14%) and ST-41/44 complex/Lineage 3 (14%). Eight new sequence types were found. None of the clonal complexes were significantly associated with increased mortality. Phenotype B:15:P1.7,16 tended to be a better predictor of death than ST-32. Although the numbers were low, the present study indicates that phenotyping may be a better predictor of mortality than MLST, which suggests that each typing method has its advantages and disadvantages. If this notion can be confirmed by other studies, it may stimulate additional research regarding the pathogenesis of severe illness, for example, if certain surface molecules trigger a cytokine storm more than others. PMID- 19133015 TI - Quinolones and oropharyngeal tularemia. PMID- 19133016 TI - Ventricular assist devices for bridge to myocardial repair. PMID- 19133017 TI - Matching efficacy of online hemodiafiltration in simple hemodialysis mode. AB - PUREMA H (referred to as PES) is an innovative dialysis membrane for enhanced low molecular-weight (LMW) protein removal. The purpose of the study was to prove whether its efficacy in hemodialysis (HD) matches that of online hemodiafiltration (HDF) with conventional high-flux membranes. In a prospective, randomized, cross-over study on eight maintenance dialysis patients, treatment efficacy of HD with PES was compared with online postdilution HDF with the two synthetic high-flux membranes polysulfone (referred to as PSU) and Polyamix (referred to as POX). Apart from the infusion of replacement fluid, which was set at 20% of the blood flow rate of 300 mL/min, operating conditions in HD and HDF were kept identical. Small solute and LMW protein plasma clearances as well as the reduction ratio (RR) of cystatin C and retinol-binding protein were not different between the therapies. HDF with POX resulted in a significantly lower myoglobin RR as compared with HD with PES, and HDF with PSU. A 4% higher beta(2) microglobulin RR was determined in HDF with PSU (73 +/- 5%) as compared with PES in HD (69 +/- 5%). The albumin loss was below 1 g for all treatments. Despite the fact that simple HD did not fully exploit the characteristics of PES, it achieved essentially similar LMW protein removal and albumin loss as compared with online postdilution HDF with the conventional synthetic high-flux membranes PSU and POX. Therefore, HD with PES may have beneficial effects on the outcome of maintenance dialysis patients similar to high-efficiency HDF. PMID- 19133018 TI - Serum free-light chain removal by high cutoff hemodialysis: optimizing removal and supportive care. AB - In multiple myeloma the predominant cause of irreversible renal failure is cast nephropathy, secondary to excess kappa or lambda serum free light chains (FLCs). These molecules are efficiently cleared by hemodialysis (HD) using the Gambro HCO 1100 dialyzer. To optimize the removal of FLCs by this dialyzer we have studied the effect of dialyzers in series, dialyzer change, and hemodiafiltration in 14 patients with multiple myeloma and renal failure. The clearance rates of both kappa FLCs and lambda FLCs were significantly increased on two dialyzers from 19 (7.3-34)-15.3 (9-28) mL/min to 47 (17-79)-35.5 (20-57) mL/min, respectively. Clearance rates of both FLCs decreased over the course of the dialysis sessions (both P < 0.001). Changing the dialyzer during a HD session increased lambda FLC clearance rates (22.5 [6-41] to 37.6 [9-52] mL/min; P < 0.001) and decreased kappa FLC clearance rates (39.6 [9-72] to 19 [8-59] mL/min; P < 0.003). Ultrafiltration during HD increased the clearance rates of kappa FLCs (R 0.52, P < 0.01) but not lambda FLCs (R -0.25; P < 0.076). Hemodiafiltration increased the clearance rates of both kappa (19 [SD 6.8] to 32 [SD 9.8] mL/min) and lambda FLCs (15 [SD 7.8] to 20 [SD 7.7] mL/min). Albumin replacement requirements for 8 h of HD increased from 12 g for a single dialyzer to 45 g for two dialyzers in series (P < 0.001). Different protocols are required to optimize the removal of kappa and lambda FLCs in patients with myeloma and renal failure. PMID- 19133019 TI - Using detachment-promoting agents for the prevention of chronic peritoneal dialysis-associated infections. AB - Biofilms are known to be responsible for chronic peritoneal dialysis (PD)-related infections. Such infections are still frequent among patients in PD. The aim of this study was to develop a new approach in the prevention of chronic PD-related infection by regular injection of specific formulations containing detachment promoting agents. A biofilm reactor system reproducing PD-like operating conditions was developed. A first set of experiments allowed the assessment of the anti-biofilm efficacy of various formulations. Then, experiments were performed for a longer duration and selected formulations were tested and compared with taurolidine. Biofilm removal was quantified by calculating the percentage of coverage reduction compared with an untreated control. A regular weekly treatment led to a 97% reduction of the surface coverage although a daily treatment with taurolidine still left 48% of the biomass on the surface. Such treatment is recommended to reduce the frequencies of chronic PD-related infections. PMID- 19133020 TI - Tissue-engineered skin containing mesenchymal stem cells improves burn wounds. AB - There is increasing evidence showing that adult stem cells are useful for tissue regeneration. Bone marrow mesenchymal stem cells (MSCs) are self-renewing and are potent in differentiating into multiple cells and tissues. To investigate the practicability of repairing burn wounds with tissue-engineered (TE) skin combined with bone MSCs, we established a burn wound model in the porcine skin. With a controlling temperature and time of the burning device to obtain different degrees of burn wounds, a deep dermal partial thickness burn was introduced to the porcine skin using a heated-brass contact injury at 100 degrees C for 20 s. Collagen-GAG scaffolds were utilized as the matrix; MSCs separated from pigs were seeded on them to form the skin equivalent. When grafted to the burn wounds, the TE skin containing MSCs showed better healing and keratinization, less wound contraction, and more vascularization. Grafts proliferated well and contributed to the neo-tissues. These data suggest that TE skin containing MSCs in a burn defect can accelerate wound healing and receive satisfactory effects. PMID- 19133021 TI - In vitro and in vivo characterization of a silk fibroin-coated polyester vascular prosthesis. AB - Silk fibroin (SF) is well known to be biocompatible, degradable, and nontoxic. In this study, SF was impregnated into a porous polyester graft (InterVascular external velour, InterVascular, Inc., La Ciotat, France), 8 mm in diameter. The SF-impregnated graft was investigated in vitro and in vivo to evaluate its potential for use as a new vascular graft impervious to blood, while retaining high porosity for tissue ingrowth and biological healing. For in vitro investigation, the water permeability, coating weight, morphology, and mechanical properties of the SF-impregnated grafts were compared with collagen-coated grafts (InterGard grafts, InterVascular, Inc.). The water permeability of the controls (1388 +/- 30.5 mL/cm(2)/min at 120 mm Hg) was reduced >99% by SF impregnation, rendering the graft impervious to blood. The coating weight of the collagen was 117 +/- 22 mg/g of graft, producing a slightly lower value than the InterGard prosthesis (302 +/- 23 mg/g). For the in vivo experiment, six SF-sealed vascular grafts were implanted in the abdominal aorta of dogs for scheduled periods ranging from 4 h to 6 months. Commercial collagen-impregnated grafts (InterGard) and untreated external velour grafts (InterVascular) were also implanted for scheduled periods ranging from 1 to 6 months for comparison. Gross observation of the explanted grafts and histological examination of the representative sections were conducted for two types of grafts using a light microscope after hematoxylin eosin staining. These SF-impregnated grafts showed less foreign body and inflammation reactions, and the SF layer was almost completely absorbed. The average of the values in each period for the SF grafts was 48% neointima at 1 month, 85% at 3 months, and 97% at 6 months, whereas those of the InterGard prostheses was 34, 46, and 90%, respectively. This study demonstrated that the use of a biodegradable SF as biological sealant can be a feasible approach to prepare impervious textile arterial prostheses. The SF-impregnated graft showed less thrombogenesis and induced host cell migration along the matrix without foreign body or inflammatory reactions. Moreover, it appears to facilitate the development of endothelial-like cells. PMID- 19133022 TI - A new method of in vitro wear assessment of the UHMWPE tibial insert in total knee replacement. AB - The wear of the ultra-high-molecular-weight polyethylene (UHMWPE) tibial insert was determined using a four-station knee simulator. The bearings were subjected to flexion/extension (between 0 and 58 degrees ), anterior-posterior translation (between 0 and -5.2 mm), internal-external rotation (between -1.9 degrees and +5.7 degrees ), and a maximum axial load of 2.6 KN, as per ISO 14243-1,2,3. The wear tests were run at a frequency of 1.1 Hz for 5 million cycles, and the wear of the inserts (n = 3) was determined using the gravimetric method. The novelty of the study was a special setup developed to simulate, as realistically as possible, in vivo conditions; this involved fixing the femoral component to the distal end of a synthetic femur model by a qualified orthopedic surgeon using an approved method. After 5 million cycles, the mean weight losses were 11.16, 19.74, and 12.61 mg for specimens #1, #2, and #3, respectively. Visual and nondestructive inspections for each of the test specimens showed similar wear tracks and these were very similar to those seen on inserts retrieved after 2 years in vivo. These results show the efficacy of the new in vitro UHMWPE wear assessment method. Furthermore, a comparison between the present in vitro results and those reported in a relevant previous study provide some insight into the influence of the method used to fix the femoral component to the simulator on the wear magnitude and patterns of the tibial insert. PMID- 19133023 TI - Oxygen delivery and hemoglobin concentration in cardiac surgery: when do we have enough? AB - For many years it has been assumed that patients undergoing cardiac surgery utilizing cardiopulmonary bypass accumulate an "oxygen debt" that requires a higher postoperative hemoglobin concentration for its reversal. Much of this evidence has now been discredited due to mathematical error with recent research suggesting critical levels of oxygen delivery are lower than previously thought. This article aims to explore the relationship between observed and critical oxygen delivery with an estimation of the minimal hemoglobin required. This was a single-center observational study. Nineteen adult elective cardiac surgery patients were recruited to participate with four subsequently excluded. Observed measurements of oxygen delivery were recorded and compared with calculated "critical" values adjusted for temperature. The hemoglobin value that represented critical oxygen delivery was compared with the observed value to identify any "hemoglobin reserve." At no perioperative time point did observed oxygen delivery or critical hemoglobin concentration significantly approach its corresponding critical value. Current transfusion practice in noncritically ill cardiac surgery patients may be considered excessive if systemic oxygen requirement is the sole parameter considered. PMID- 19133024 TI - Changes in atrial natriuretic peptide levels during cardiac bypass in the fetal goat. AB - The aim of this study is to evaluate the effect of fetal cardiac bypass on the production and secretion of fetal atrial natriuretic peptide (ANP) in the goat. Eighteen pregnant goats, at days 120 to 140 of gestation, were randomly divided into control (n = 8) and bypass (n = 10) groups. The control group underwent a sham procedure involving fetal sternotomy and cannulation. The bypass group underwent fetal cardiac bypass using a centrifugal pump for 30 min. Fetuses in the bypass group exhibited hypoxia, hypercapnia, and acidosis during and after cardiac bypass. The pulse index (PI) of the umbilical artery in the bypass group increased significantly after cardiac bypass compared with the control group. Tei indices of the left and right ventricles in the bypass group increased remarkably after cardiac bypass. Plasma troponin I levels in the bypass group increased significantly compared with that of the control group. Plasma ANP levels increased markedly in the bypass group after cardiac bypass, and the difference between two groups was significant. Transcriptional levels of ANP mRNA in the fetal heart elevated remarkably in the bypass group compared with the control group at 2 h after the bypass. A significant positive correlation between plasma ANP levels and Tei indices of the ventricles, plasma troponin I was observed (left ventricular Tei index, r = 0.606, P < 0.01; right ventricular Tei index, r = 0.581, P < 0.01; plasma troponin I, r = 0.275, P < 0.05). In conclusion, fetal cardiac bypass promoted the production and secretion of ANP and was associated with fetal cardiac dysfunction. PMID- 19133025 TI - Score model for the evaluation of dialysis membrane hemocompatibility. AB - The interaction of blood with artificial surfaces is of particular interest during hemodialysis treatments with extracorporeal blood circuits. Components of the extracorporeal blood circuit are known to have only a moderate, sometimes even an unfavorable hemocompatibility, and thus may provoke adverse biochemical or clinical sequelae. This article describes a newly established hemocompatibility assessment score. This score is based on on a standardized series of in vitro tests and is applied to commercially available hemodialysis membranes. It relates to a variety of membrane polymers, such as regenerated cellulose, diethylaminoethyl-modified cellulose, polyethersulfone/polyarylate blends and polysulfone. In order to compare different polymers used in the manufacturing of dialysis membranes, a set of the following hemocompatibility parameters was assessed and assembled to an overall score: generation of complement factor 5a, thrombin-antithrombin III-complex, release of platelet factor 4, generation and release of elastase from polymorphonuclear granulocytes, and platelet count. With respect to these parameters, the results reveal major differences between the selected dialysis membranes. This new score model proves to be an efficient tool to derive objective results, and it may, thus, be used in the future to facilitate the selection of membrane polymers with an appropriate hemocompatibility pattern for dialysis therapy. PMID- 19133026 TI - Subcutaneous transposition of the superficial femoral artery for arterioarterial hemodialysis: technique and results. AB - We report the use of subcutaneous transposition of the femoral artery (STFA) for placement of both inflow and outflow needles in 14 hemodialysis (HD) adult patients with difficult access. Follow-up time was 318 months during which a total of 3215 arterioarterial HD sessions were done. Kt/V values ranged between 0.71 and 1.59. Elevated access recirculation and dialysis outflow pressures were common findings to all patients. Complications were: (i) two episodes of bleeding secondary to puncture-related arterial wall laceration, repaired by stitching; (ii) three episodes of thrombosis in two patients, all successfully declotted; (iii) three puncture-related complications needing placement of a vein interposition graft, namely, aneurysm, pseudoaneurysm, and arterial stenosis; and (iv) one case of arterial ligation because of suppurative puncture site infection, without subsequent distal ischemia signs or claudication. The use of STFA should only be reserved for patients in urgent need for vascular access with no remaining options. PMID- 19133027 TI - Acid-base balance and oxygen tension during dialysis in uremic patients with chronic obstructive pulmonary disease. AB - Recent reports on the effects of dialysis on acid-base balance and metabolic acidosis correction in end-stage renal disease (ESRD) patients with chronic obstructive pulmonary disease (COPD) are lacking. Here, we compared acid-base balance and blood gasses among 14 patients with established COPD (group A) and eight patients with normal respiratory function (group B). The two groups were homogeneous for age, time on dialysis, and male/female ratio. At the beginning of dialysis, acid-base balance and blood gasses were comparable between patients of groups A and B. A significant difference between groups was observed only in pCO(2) at 20 min, together with a delay in pH increase. Effective correction of acidosis was reported at the end of dialysis and is not significantly affected by COPD. Nevertheless, weight loss must be carefully monitored in these patients in order to prevent hyperhydration and worsening of respiratory function. PMID- 19133028 TI - Endothelial cell retention on a viscoelastic nanocomposite vascular conduit is improved by exposure to shear stress preconditioning prior to physiological flow. AB - In this study, endothelial cell (EC)-seeded nanocomposite grafts were preconditioned with 1-2 dynes/cm(2) in vitro to establish whether low shear stress resulted in improved cell adherence prior to physiological shear stress (15 dynes/cm(2)). Alamar blue cell viability was assessed. Polymerase chain reaction was conducted for glyceraldehyde-3-phosphate dehydrogenase, transforming growth factor beta-1 (TGFbeta-1), vascular endothelial growth factor receptor-1 (VEGFR-1), platelet EC adhesion molecule-1, and vascular endothelial growth factor receptor-2 (VEGFR-2). The Alamar blue results demonstrated improved cellular retention following preconditioning (P < 0.001). VEGFR-2 and TGFbeta-1 expression was up-regulated, and VEGFR-1 down-regulated following preconditioning. This investigation confirms previous findings regarding the potential benefits of preconditioning, and demonstrates that these benefits can be applied to ECs seeded on the nanocomposite employed. It also demonstrates further the suitability and potential of nanocomposite for future use in tissue engineered cardiovascular devices. PMID- 19133029 TI - Effect of RGD-immobilized dual-pore poly(L-lactic acid) scaffolds on chondrocyte proliferation and extracellular matrix production. AB - Interactions between cell and polymer surface have great implications in tissue engineering. In this study, chondrocyte proliferation and matrix production were examined using porous poly(L-lactide) (PLLA) scaffolds that have different surface characteristics. PLLA scaffolds were prepared using a gas-foaming method, and subjected to surface modifications through plasma treatment and subsequent in situ grafting of hydrophilic acrylic acid (AA). To immobilize peptide ligands, the AA-grafted PLLA scaffolds (PLLA-PAA) were further reacted with either Gly-Arg Asp-Gly (GRDG) or Gly-Arg-Gly-Asp (GRGD) to produce PLLA-PAA-GRDG or PLLA-PAA GRGD scaffold, respectively. The average porosities of the scaffolds were more than 90%, and their pore sizes ranged from 200 approximately 300 to 10 approximately 50 microm for large and small pores, respectively. The concentrations of each bound component were 2.14 x 10(-4) mmol/cm(2) for AA, 1.87 nmol/g for GRDG, and 1.68 nmol/g for GRGD. When chondrocytes were seeded onto the different PLLA scaffolds, cell adhesion and proliferation were highly affected as the substrate types vary. The RGD-immobilized scaffolds resulted in higher cellularity and better accumulation of total glycosaminoglycan than the others. Histological staining of Safranin O showed that the deposited extracellular matrix was more intense and widely distributed in the PLLA-PAA-GRGD scaffold. The present data suggest that immobilization of RGD peptide on the AA-grafted PLLA scaffold can be an effective tool for chondrocyte attachment and proliferation, and that it may also be helpful to facilitate cartilaginous tissue formation. PMID- 19133030 TI - The use of an approved biodegradable polymer scaffold as a solid support system for improvement of islet engraftment. AB - The aim of this study was to investigate whether the use of a medically approved biodegradable scaffold as a solid support system would enhance graft survival following transplantation into the omental pouch in a preclinical large animal model. Six beagle dogs underwent total pancreatectomy followed by islet autotransplantation into the omental pouch. Four dogs received islets seeded in a biodegradable polymer scaffold and two received free islets without a scaffold. All four animals that received islets in the scaffold became normoglycemic without exogeneous insulin injection. One dog, transplanted with the largest number of islets, maintained a normal metabolic state until the graft was removed at 5 months posttransplant. In two out of the three that received a marginal islet mass, insulin independence was sustained up to 2 months. In contrast, two dogs transplanted with a similar marginal mass without the scaffold never became normoglycemic. Histological examination of the grafts in the scaffold showed numerous well-granulated, insulin-containing cells as well as glucagon-positive cells. These results indicate that biodegradable scaffolds may enhance survival and function of islet grafts. Manipulation of the microenvironment of transplanted islets may constitute the basis for new approaches to enhance islet engraftment. PMID- 19133031 TI - Temperature control using a heat exchanger of a cardioplegic system in cardiopulmonary bypass model for rats. AB - Small animal cardiopulmonary bypass (CPB) model would be a valuable tool for investigating pathophysiological and therapeutic strategies on bypass. However, the rat CPB models have a number of technical limitations. Effective maintenance and control of core temperature by heat exchanger (HE) is among them. The purpose of this study was to confirm the effect of rectal temperature maintenance and hypothermic control using a HE of cardioplegia system in CPB model for rats. The miniature circuit consisted of a reservoir, HE, membrane oxygenator, and roller pump; the static priming volume was 40 cc. In the first stage of experiment, 10 male Sprague-Dawley rats were divided into two groups; HE group was subjected to CPB with HE from a cardioplegia system, and control group was subjected to CPB with warm water circulating around the reservoir. Partial CPB was conducted at a flow rate of 40 mg/kg/min for 20 min after venous cannulation (via the internal jugular vein) and arterial cannulation (via the femoral artery). Rectal temperature was measured after anesthetic induction, after cannulation, 5, 10, 15, and 20 min after CPB. Arterial blood gas with hematocrit was also analyzed, 5 and 15 min after CPB. In the second stage with the same experimental setting, rectal temperatures were lowered in 10 rats to the target temperature of 32 degrees C. After reaching the target temperature, animals were rewarmed. Rectal temperature was measured after cannulation, 5, 10, 15, 20, 25, and 30 min after CPB. Arterial blood gas with hematocrit was also analyzed, 5 and 15 min after CPB. Rectal temperature change differed between the two groups (P < 0.01). The temperatures of the HE group were well maintained during CPB, whereas the control group was under progressive hypothermia. Rectal temperature 20 min after CPB was 36.16 +/- 0.32 degrees C in the HE group and 34.22 +/- 0.36 degrees C in the control group. In the second set of experiments, the hypothermia targeted (32 degrees C) was reached in 15 min (from 35.56 +/- 1.05 degrees C to 31.75 +/- 0.47 degrees C). The rats were successfully rewarmed within the observation period of 30 min. Arterial blood gases and hematocrits showed no further significant findings. We confirmed the effect of rectal temperature maintenance and hypothermic control using an HE of cardioplegia system in CPB model for rats. This model would be a valuable tool for further use in hypothermic CPB experiments in rats. PMID- 19133033 TI - Innovative, replaceable heart valve: concept, in vitro study, and acute in vivo study. PMID- 19133034 TI - June 2008 issue: Jesus Herreros from the University Hospital of Navarra, Pamplona, Spain. Letter to the editor. PMID- 19133036 TI - Indigenous women's health in the spotlight. PMID- 19133037 TI - Aboriginal and Torres Strait Islander women's health: acting now for a healthy future. AB - This paper summarises the recent RANZCOG Indigenous Women's Health Meeting with recommendations on how the College and its membership can act now to improve the health of Aboriginal and Torres Strait Islander women and infants. PMID- 19133038 TI - First trimester predictors of adverse pregnancy outcomes. AB - AIM: To identify first trimester indicators of adverse pregnancy outcomes. METHOD: Data were obtained from the statewide evaluation of first trimester screening for Down syndrome in Western Australia which included 22,695 pregnancies screened between August 2001 and October 2003. Screening data were linked with pregnancy outcome information from the Hospital Morbidity Database and the Birth Defects Registry. The odds ratios (OR) of adverse outcomes were analysed for combined risk incorporating maternal age, nuchal translucency (NT) and biochemical parameters and then separately for each parameter (pregnancy associated plasma protein-A (PAPP-A), free beta human chorionic gonadotropin (beta-hCG) and NT). RESULTS: Risk assessments for first trimester combined screening are derived from maternal age, ultrasound measurement of fetal NT, maternal serum free beta-hCG and PAPP-A. Increased combined risk for Down syndrome was significantly (P < 0.01) associated with spontaneous loss at or before 24 weeks gestation (OR 13.51), birth defects (OR 6.58) and preterm birth at or before 32 weeks gestation (OR 3.2). Maternal serum PAPP-A below the 5th centile was associated with Down syndrome (OR 8.43), spontaneous loss before 24 weeks (OR 5.04) and later than 24 weeks (OR 4.50), preterm delivery before 32 weeks (OR 3.11) and before 37 weeks (OR 2.24). NT above the 95th centile was associated with Down syndrome (OR 43.91), birth defects (OR 4.02) and spontaneous loss before 24 weeks (OR 6.24). Low levels of free beta-hCG and increased NT were less consistently associated with adverse outcomes and high levels of free beta hCG showed limited use as an indicator. The detection rates for all outcomes other than Down syndrome were less than 40%. CONCLUSION: Biochemical indicators and NT that are measured during first trimester screening for Down syndrome show a number of associations with adverse outcomes, but do not show appropriate performance characteristics for screening tests. These data are consistent with the view that the individual components, specifically low PAPP-A levels alone, do not provide an effective screening tool for adverse pregnancy outcomes. PMID- 19133039 TI - The prevalence of inherited thrombophilic polymorphisms in an asymptomatic Australian antenatal population. AB - AIM: Inherited thrombophilic polymorphisms have been linked to pregnancy-related thromboembolism and other adverse pregnancy outcomes. As there are limited data on the prevalence of these polymorphisms in Australian populations, we aimed to assess this in an antenatal population. METHODS: Healthy nulliparous women (n = 2031) were recruited to this study. The women had no past or family history of venous thromboembolism. Women were excluded if they or a family member was known to be a carrier of any thrombophilic marker. Genotyping from venous blood for the factor V Leiden, prothrombin 20210A, MTHFR 677 and 1298 and thrombomodulin C1418T polymorphisms was undertaken. RESULTS: Key findings were that 107 of 2019 (5.30, 95% confidence interval 4.36-6.37%) women tested were heterozygous carriers of factor V Leiden and one was homozygous (0.05, 0-0.27%); 2.43% of women were heterozygous carriers of the prothrombin gene mutation (1.80-3.20%) while no women were homozygous for this mutation; 11.62% (10.22-13.02%) and 9.98% (8.67 11.29%) were homozygous for the MTHFR 677 and 1298 polymorphisms, respectively, and 3.43% (2.63-4.22%) of women were homozygous for the thrombomodulin polymorphism. CONCLUSIONS: The prevalence of these polymorphisms is consistent with previously published data in Caucasian populations. These data will provide the basis for further assessment of the relationship between poor pregnancy outcome and these inherited thrombophilic polymorphisms in an asymptomatic antenatal population. PMID- 19133040 TI - Willingness of pregnant women and clinicians to participate in a hypothetical randomised controlled trial comparing vaginal delivery and elective caesarean section. AB - BACKGROUND: Elective caesarean section is controversial in the absence of compelling evidence of the relative benefits and harms compared with vaginal delivery. A randomised trial of the two procedures to compare outcomes for women and babies would provide the best quality scientific evidence to confirm this debate but it is not known whether such a trial would be feasible. AIMS: To ascertain the proportion of primiparas and clinicians who would participate in a hypothetical randomised controlled trial comparing vaginal delivery with elective caesarean section. METHODS: Pregnant women (mean 22 weeks gestation) recruited from public and private antenatal clinics at a major tertiary referral centre were interviewed to ascertain their willingness to participate in a hypothetical randomised controlled trial. A self-administered questionnaire was mailed to midwives, obstetricians, urogynaecologists and colorectal surgeons, and results between groups were compared. RESULTS: One hundred pregnant women, 84 midwives, 166 obstetricians, 12 urogynaecologists and 87 colorectal surgeons participated. Only 14% (95% confidence interval (CI), 8-22) of pregnant women and 31% (95% CI, 26-36) of clinicians indicated that they would participate in a randomised controlled trial. CONCLUSIONS: A randomised controlled trial comparing vaginal delivery and elective caesarean section may not be feasible due to low levels of willingness to participate, particularly among pregnant women. PMID- 19133041 TI - Tumour necrosis factor (-308A) polymorphism in pre-eclampsia: meta-analysis of 16 case-control studies. AB - BACKGROUND: The guanine to adenine transition at position -308 nucleotides in the tumour necrosis factor promoter region (TNF -308A) is a putative genetic risk factor for pre-eclampsia/eclampsia (PE/E). However, association studies have reported conflicting findings and have generally been underpowered to exclude modest effect sizes. AIM: To assess and synthesise the available evidence for the association of the TNF (-308A) polymorphism with PE/E. METHODS: Systematic review and random effects meta-analysis of genetic association studies. RESULTS: Sixteen eligible case-control studies in which a total of 1919 patients with PE/E and 2374 controls participated were identified. The studies were generally small but of reasonable methodological quality. Random effects meta-analysis improved the precision of the estimate of effect size but did not detect a statistically significant association: pooled relative risk 1.02 (95% confidence interval 0.86 to 1.20). CONCLUSIONS: The available data are not consistent with more than a very modest association between the TNF (-308A) allele and PE/E. Screening pregnant women for this allele in order to guide antenatal surveillance or treatment is unwarranted. Future research efforts should focus on alternative candidate genes. PMID- 19133042 TI - Implementation of a clinical practice guideline for smoking cessation in a public antenatal care setting. AB - BACKGROUND: Despite high level evidence showing that antenatal smoking cessation programs are effective in reducing the number of women who smoke during pregnancy and the number of low birthweight and preterm births, few Australian hospitals have adopted a systematic approach to assist pregnant women to stop smoking. AIMS: The aim of this study was to assess the effectiveness of a smoking cessation guideline, developed specifically for clinicians providing antenatal care in public maternity hospitals, combined with an implementation program on the uptake of evidence-based practice. METHODS: A clinical practice guideline was developed and an implementation strategy was tested, using a prospective before and-after study design, at the Mater Mothers' Hospital in Brisbane. Women were surveyed in late pregnancy, pre- and post-implementation. The primary outcome measures were women's report of appropriate smoking cessation support received, specifically, information brochures and referral to Quitline. Secondary outcome measures included women's report of smoking status in late pregnancy and relapse rates. RESULTS: Post-implementation, more women reported receiving written materials on smoking cessation (76% vs 35%; relative risk (RR) 3.4; 95% confidence interval (CI) 2.7, 4.2) and referral to Quitline (67% vs 14%; RR 4.9; 95% CI 3.0, 8.0). While not statistically significant, fewer women post implementation reported smoking in late pregnancy (19.5% vs 16.7%) and fewer reported smoking > 10 cigarettes per day (38% vs 25%). CONCLUSIONS: Clinical practice guidelines specifically designed for a public maternity care setting combined with an implementation program resulted in an increase in evidence-based practice with some indication of improved smoking behaviour for women. PMID- 19133043 TI - Effectiveness of prenatal diagnosis of congenital heart defects in South Australia: a population analysis 1999-2003. AB - AIMS: To report on the efficiency of fetal echocardiography, outcome after prenatal diagnosis of congenital heart disease (CHD) and the effectiveness of South Australia's obstetric screening program in detecting CHD. METHODS: A retrospective audit of our institutional cardiac database, the South Australia Birth Defects Register and the South Australia Pregnancy Outcome Unit from 1999 to 2003. RESULTS: CHD was present in 10.7 per 1000 births and was potentially detectable in 5.2 per 1000 births. Of these, only 22.5% were referred for fetal echocardiography. Prenatal detection of transposition of the great arteries occurred in four of 27 (14.8%), atrioventricular septal defect in five of 21 (23.8%), tetralogy of Fallot in seven of 28 (25.0%) and hypoplastic left heart syndrome in 20 of 33 (60.6%). Fetal echocardiography had 95.2% sensitivity, 99.5% specificity, 99.0% positive predictive value and 97.6% negative predictive value for CHD. A prenatal diagnosis of CHD resulted in termination in 30.1%, stillbirth in 7.8% and neonatal death in 3.9% of cases. CONCLUSION: Routine obstetric ultrasound detects CHD infrequently. Outflow tract lesions are most commonly missed. Fetal echocardiography is an accurate tool for prenatal diagnosis of CHD. Pregnancy outcomes after prenatal diagnosis are comparable to other Australasian and western countries, suggesting uniformity in parental counselling and population behaviour. Strategies that improve routine obstetric ultrasound detection of CHD are likely to yield the greatest impact. PMID- 19133044 TI - Prepartum and intrapartum caesarean section rates at Mater Mothers' Hospital Brisbane 1997-2005. AB - AIM: To document the rise in prepartum and intrapartum caesarean section and the demographic and medical factors contributing to the rise. METHODS: Data from 52,423 deliveries between January 1997 to May 2005 were analysed for yearly change in caesarean section rates and multiple demographic and medical factors. RESULTS: The prepartum caesarean section rate increased by 1.6% per year and the intrapartum caesarean section rate by 0.8% per year. There was no increase in the overall prevalence of obesity, short stature, advanced maternal age, medical complications or previous caesarean section. There were significant increases in nulliparity, private care, induction of labour and the use of electronic monitoring, but these were insufficient to explain the magnitude of the rise. CONCLUSION: The increase in prepartum and intrapartum caesarean section displayed was not fully explained by medical and demographic changes in the population. PMID- 19133045 TI - A randomised controlled trial of uterine exteriorisation and non-exteriorisation at caesarean section. AB - BACKGROUND: There is not enough information to evaluate the routine use of exteriorisation of the uterus for repair of the uterine incision and further studies are needed to examine the value of exteriorisation of the uterus at caesarean section as against non-exteriorisation. AIMS: To assess intraoperative and postoperative morbidity following exteriorisation of the uterus at caesarean section as compared to those with non-exteriorisation. METHODS: A randomised controlled trial at Obafemi Awolowo University Teaching Hospital, Ile Ife, Nigeria. Two hundred and ten women were randomised to either exteriorisation (N = 105) or non-exteriorisation group (N = 105). Statistical analysis using SPSS compared intraoperative and postoperative outcome. The outcome measures include intraoperative blood loss, number of analgesic dose, difference in operating time, febrile morbidity, intraoperative complications, and duration of hospital stay. RESULTS: There were significant reductions in intraoperative blood loss (P < 0.05), number of analgesic dose (P < 0.05) and duration of hospital stay (P < 0.05) in the uterine exteriorisation group as compared to those in the non exteriorisation group. There was no significant difference in operating time, febrile morbidity, intraoperative complications and operating time. The period for return of bowel function was significantly longer in the exteriorisation group. CONCLUSION: Exteriorisation of uterus at caesarean section is associated with less intraoperative blood loss, less number of analgesic dose and shorter hospital stay. PMID- 19133046 TI - Transplacental haemorrhage may explain the intrapartum transmission of HIV. A pilot study uses flow cytometry to quantify maternal red blood cells in infants born vaginally or by caesarean section. AB - Intrapartum transmission is epidemiologically important for some viruses such as HIV and hepatitis B virus, but its precise mechanism is unknown. We hypothesised that the ability of elective caesarean section to prevent HIV may be due to prevention of transplacental microtransfusions of blood during labour. Their frequency is not known so we performed a pilot study which showed evidence of transplacental transfusion from mother to fetus in one of ten mother-infant pairs delivering vaginally and none of ten delivering by elective caesarean section. We conclude that transplacental transfusion occurs and is one possible mechanism for the intrapartum transmission of viruses from mother to baby. PMID- 19133047 TI - Peripartum hysterectomy in Aba southeastern Nigeria. AB - Emergency peripartum hysterectomy is a challenging but life-saving procedure. In this descriptive study carried out in a rural Nigerian hospital, we found an incidence of emergency peripartum hysterectomy of 5.4 per 1000 deliveries and a significant association with abdominal mode of delivery, unbooked status, previous caesarean section and placenta previa. The most common indications for peripartum hysterectomy were placenta accreta (47.6%) and uterine rupture (28.6%). There were five (23.8%) maternal deaths and other complications included sepsis (five), bladder injury (three) and prolonged hospital stay (11). PMID- 19133048 TI - Monozygotic triplets after single blastocyst transfer: case report and literature review. AB - A 28-year-old woman undertaking in vitro fertilisation had a single day five blastocyst transferred, resulting in a monozygotic triplet pregnancy. This case report illustrates the potential for multiple pregnancies even after transfer of a single embryo. A literature review regarding monozygotic triplets following use of assisted reproductive techniques reveals that an unexpectedly large proportion of these cases are associated with blastocyst transfer. PMID- 19133049 TI - Prolapse worsens with age, doesn't it? AB - BACKGROUND: Female pelvic organ prolapse is common and generally thought to worsen over time. This assumption has recently become less plausible, as the author and others have been able to show that mild to moderate pelvic organ descent is common in young, nulligravid women. AIMS: To investigate the relationship between age and pelvic organ prolapse. METHODS: The records of 1110 women seen for interview, clinical examination, urodynamics and ultrasound were evaluated in a retrospective study. Data were analysed to investigate the relationship between patient age at presentation and pelvic organ descent on clinical examination and ultrasound imaging. RESULTS: After removal of 139 datasets of women with previous incontinence or prolapse surgery, 971 datasets remained. Mean age was 54 years (17-90), mean vaginal parity was 2.4 (0-12), with 31% complaining of prolapse. We found weak complex relationships between age and cystocele/rectocele staging, with a positive correlation to menopause and a negative relationship thereafter. This was confirmed on imaging findings, with regression showing an almost parabolic fitted line plot for cystocele and rectocele, but a near-linear curve for uterine prolapse. In nulliparous women, the positive relationship between age and cystocele in premenopausal women was still significant (P = 0.028), indicating that it is not explained by the confounding effect of child bearing. CONCLUSIONS: Ageing seems to play a complex role in the aetiology and pathogenesis of pelvic organ prolapse. Our results contradict epidemiological studies showing age to be a major risk factor for pelvic reconstructive surgery and pelvic organ prolapse. PMID- 19133050 TI - Quality-initiated prophylactic antibiotic use in laparoscopic-assisted vaginal hysterectomy. AB - BACKGROUND: An evidence-based initiation of quality improvement activity for reducing the usage of prophylactic antibiotic in laparoscopic-assisted vaginal hysterectomy (LAVH) in a tertiary hospital. AIMS: The authors investigated whether single or multiple doses of cefazoline were more cost-effective in preventing postoperative infection associated with LAVH. METHODS: The study groups comprised of 310 patients who had undergone LAVH continuously in a medical centre. Patients were divided into two groups on the basis of whether they received a single dose or multiple doses (range: two to four doses) of cefazoline during the perioperative period. Postoperative infections such as pelvic cellulitis or abscess, vaginal cuff abscess, wound infection and urinary tract infection that occurred either during hospitalisation or within one month after discharge were observed and recorded. Incremental cost-effectiveness ratio (ICER) was calculated using the mean direct drug cost and the prophylactic effect of infection in both groups. RESULTS: The prophylactic effect of infection was similar in the single-dose group and the multiple-dose group (94.6% vs 93.9%, P = 0.986). The ICER was significantly lower in the single-dose group (153.3 vs 460.4, P < 0.001). CONCLUSIONS: The result revealed that a single dose of cefazoline is more cost-effective than multiple doses in the prevention of infection associated with LAVH. It fulfils the goal of cost minimisation and quality of care in today's environment of medical cost containment. PMID- 19133051 TI - The diagnosis and treatment of Mullerian adenosarcoma of the uterus. AB - BACKGROUND: Adenosarcoma of the uterus is one of the rare types of gynaecological malignant tumours. Poor awareness of it among clinicians makes it difficult to diagnose correctly and timely, so it is helpful to enrich and update our knowledge about the tumour with new information of patients. AIM: To improve the level of diagnosis and treatment of Mullerian adenosarcoma of the uterus. METHODS: The medical data of nine patients with Mullerian adenosarcoma of the uterus who were treated from May 1995 to March 2006 in our hospital were analysed retrospectively. The analysis focused on clinicopathological features, treatment and prognosis. RESULTS: Patients typically presented with abnormal uterine bleeding, pain in the lower abdomen, enlargement of the uterus, a mass in the uterine cavity and/or a cervical neoplasm. Microscopically, the glands were lined by benign or atypical glandular epithelium, together with sarcomatous stromal cells which showed characteristic structures of 'periglandular cuff' of increased cellularity and 'intraglandular polypoid projections'. The primary diagnostic rate was 33.3% and the average interval from symptom onset to final diagnosis was 13 months and eight weeks for pre- and postmenopausal patients, respectively. The prognosis was better in early stage disease in young patients. CONCLUSIONS: The most common symptom of adenosarcoma of the uterus is abnormal uterine bleeding. Younger patients are more often misdiagnosed. Clinical stage and age of the patient are closely related to prognosis. Surgery is the primary treatment, and chemotherapy may be somewhat beneficial. PMID- 19133052 TI - Gamete donors' and recipients' evaluation of donor counselling: a prospective longitudinal cohort study. AB - BACKGROUND: Those considering donating or receiving gametes are required to undergo counselling to ensure that they are aware of and consider the psychosocial, emotional and ethical complexities of donor conception before proceeding. AIMS: The aim of this study was to evaluate gamete donors' and recipients' views about donor counselling and beliefs about disclosure of the use of donor gametes to conceive, before and after attending counselling. METHOD: All donors and recipients and their partners who attended donor counselling at Monash IVF, Melbourne, Australia, for the first time between February 2003 and March 2004 were asked to complete self-report questionnaires. Using seven-point Likert scales where higher scores indicated more positive views, participants rated the importance of discussing a range of topics before, and the usefulness of discussing these after attending counselling. RESULTS: Pre-counselling questionnaires were completed by 152 of 184 (83%) individuals and 72 (47%) of those also completed the post-counselling questionnaire. Donors' post-counselling ratings were significantly higher than their pre-counselling ratings. For recipients the pre- to post-counselling changes were less pronounced. Both donors and recipients rated matters relating to disclosure and the possible future interaction between donor and child the most useful to discuss. More participants favoured disclosure after than before counselling (84% vs 66%, P = 0.01). CONCLUSIONS: This study suggests that counselling is beneficial for those contemplating donor procedures, particularly donors, and that most have a positive attitude towards disclosure after counselling. PMID- 19133053 TI - Primary psoas muscle abscess in pregnancy. AB - Primary iliacus-psoas muscle abscess formation is very uncommon during pregnancy. We present a case of a primary iliacus-psoas muscle abscess in pregnancy causing back pain with delayed diagnosis and treatment. Understanding the clinical presentation of iliacus-psoas muscle abscess helps with considering it in the differential diagnosis of back pain during pregnancy. PMID- 19133054 TI - Successful pregnancy outcome in a patient with moyamoya disease. PMID- 19133055 TI - Re: alcohol and pregnancy: the pivotal role of the obstetrician. PMID- 19133056 TI - Looking back: editors' pick of 2008. PMID- 19133058 TI - Effect of naproxen on the hypothalamic-pituitary-adrenal axis in healthy volunteers. AB - AIM: To study the effect of the nonsteroidal anti-inflammatory drug naproxen on the activity of the hypothalamic-pituitary-adrenal (HPA) axis in healthy volunteers. METHODS: A double-blind, randomized study in two groups of 20 healthy volunteers was performed. The activity of the HPA axis was measured before and after the use of naproxen or placebo during a period of 2 weeks. Basal plasma adrenocorticotropic hormone (ACTH) and cortisol, 24-h urinary cortisol, and circadian cortisol rhythm in saliva were determined. Plasma ACTH and cortisol were also measured during submaximal physical exercise. RESULTS: There were no significant differences between the placebo and naproxen groups in basal plasma ACTH [09.00 h 3.1 pmol l(-1), 95% confidence interval (CI) 2.0, 4.2, and 2.8 pmol l(-1), 95% CI 1.9, 3.7, respectively], cortisol levels (09.00 h 0.45 micromol l( 1), 95% CI 0.39, 0.51, and 0.40 micromol l(-1), 95% CI 0.35, 0.44, respectively), 24 h urinary cortisol excretion (67.5 nmol 24 h(-1), 95% CI 54.3, 80.7, and 86.8 nmol 24 h(-1), 95% CI 54.4, 119.2, respectively), circadian cortisol rhythm measured in salivary samples, or ACTH and cortisol concentrations after physical exercise. After the use of placebo or naproxen for 2 weeks, no significant change in any of the parameters occurred (ACTH 09.00 h 3.0 pmol l(-1), 95% CI 2.0, 3.9, and 3.0 pmol l(-1), 95% CI 2.2, 3.8, respectively; cortisol 09.00 h 0.45 micromol l(-1), 95% CI 0.37, 0.52, and 0.39 micromol l(-1), 95% CI 0.34, 0.44, respectively; cortisol urine 79.5 nmol 24 h(-1), 95% CI 59.5, 99.4, and 81.7 nmol 24 h(-1), 95% CI 64.0, 99.4, respectively), and no significant differences were found in these parameters between the placebo and naproxen groups. CONCLUSIONS: The use of naproxen does not influence the activity of the HPA axis in healthy volunteers under basal circumstances or in response to physical stress. PMID- 19133059 TI - Contribution of the activities of CYP3A, CYP2D6, CYP1A2 and other potential covariates to the disposition of methadone in patients undergoing methadone maintenance treatment. AB - AIMS: To investigate the influence of different cytochrome P450 (CYP) activities and other potential covariates on the disposition of methadone in patients on methadone maintenance therapy (MMT). METHODS: Eighty-eight patients (58 male; 21 55 years; 84 White) on MMT were studied. CYP2D6 activity [3 h plasma metabolic ratio of dextromethorphan (DEX) to dextrorphan (DOR)] was determined in 44 patients (29 male; 24-55 years), CYP1A2 activity (salivary caffeine elimination half-life) in 44 patients (21 male; 24-55 years) and CYP3A activity (oral clearance of midazolam) in 49 patients (33 male; 23-55 years). Data on all three CYPs were obtained from 32 subjects. Total plasma concentrations of (RS) methadone and total and unbound plasma concentrations of both enantiomers were measured by LC/MS. Population pharmacokinetics and subsequent multiple regression analysis were used to calculate methadone oral clearance and to identify its covariates. RESULTS: Between 61 and 68% of the overall variation in total plasma trough concentrations of (RS)-, (R)- and (S)-methadone was explained by methadone dose, duration of addiction before starting MMT, CYP3A activity and illicit morphine use. CYP3A activity explained 22, 16, 15 and 23% of the variation in unbound (R)-, unbound (S)-, total (RS)- and total (S)-methadone clearances, respectively. Neither CYP2D6 nor CYP1A2 activity was related to methadone disposition. CONCLUSIONS: CYP3A activity has a modest influence on methadone disposition. Inhibitors and inducers of this enzyme should be monitored in patients taking methadone. PMID- 19133060 TI - High-dose antipsychotic use in schizophrenia: a comparison between the 2001 and 2004 Research on East Asia Psychotropic Prescription (REAP) studies. AB - AIMS: We aimed to examine the frequency of high-dose (defined as mean chlorpromazine mg equivalent doses above 1000) antipsychotic prescriptions in schizophrenia and their clinical correlates in the context of a comparison between studies in 2001 and 2004 within six East Asian countries and territories. METHODS: Prescriptions of high-dose antipsychotic for a sample of 2136 patients with schizophrenia from six countries and territories (mainland China, Hong Kong, Korea, Japan, Taiwan and Singapore) were evaluated in 2004 and compared with data obtained for 2399 patients in 2001. RESULTS: Overall, the comparison between 2001 and 2004 showed a significant decrease in high-dose antipsychotic use from 17.9 to 6.5% [odds ratio (OR) 0.32, 95% confidence interval (CI) 0.26, 0.39, P < 0.001]. Patients who received high-dose antipsychotics were significantly more likely to have multiple admissions (OR 1.96, 95% CI 1.16, 3.33, P = 0.009), more positive psychotic symptoms such as delusions (OR 2.05, 95% CI 1.38, 3.05, P < 0.001) and hallucinations (OR 1.85, 95% CI 1.30, 2.64, P = 0.001), but less likely to have negative symptoms (OR 0.58, 95% CI 0.40, 0.82, P = 0.002). Multivariate regression analyses revealed that prescription of high-dose antipsychotics was also predicted by younger age (P < 0.001), time period of study (2001; P < 0.001), use of first-generation antipsychotic (P < 0.001) and depot antipsychotics (P < 0.001) as well as antipsychotic polytherapy (P < 0.001). CONCLUSIONS: We identified the clinical profile and treatment characteristics of patients who are at risk of receiving high antipsychotic doses. These findings should provide impetus for clinicians to constantly monitor the drug regimes and to foster rational, evidence-based prescribing practices. PMID- 19133057 TI - Biomarkers for the effects of cannabis and THC in healthy volunteers. AB - An increasing number of novel therapeutic agents are targeted at cannabinoid receptors. Drug development programmes of new cannabinoid drugs may be facilitated by the identification of useful biomarkers. This systemic literature review aims to assess the usefulness of direct biomarkers for the effects of cannabis and tetrahydrocannabinol (THC) in healthy volunteers. One hundred and sixty-five useful articles were found that investigated the acute effects of cannabis or THC on the central nervous system (CNS) and heart rate in healthy volunteers. Three hundred and eighteen tests (or test variants) were grouped in test clusters and functional domains, to allow their evaluation as a useful biomarker and to study their dose-response effects. Cannabis/THC affected a wide range of CNS domains. In addition to heart rate, subjective effects were the most reliable biomarkers, showing significant responses to cannabis in almost all studies. Some CNS domains showed indications of depression at lower and stimulation at higher doses. Subjective effects and heart rate are currently the most reliable biomarkers to study the effect of cannabis. Cannabis affects most CNS domains, but too many different CNS tests are used to quantify the drug response relationships reliably. Test standardization, particularly in motor and memory domains, may reveal additional biomarkers. PMID- 19133061 TI - The CADEUS study: burden of nonsteroidal anti-inflammatory drug (NSAID) utilization for musculoskeletal disorders in blue collar workers. AB - AIM: The aim of this study was to compare patterns of utilization of NSAIDs for musculoskeletal disorders (MSD) by occupation in a general employed population. METHODS: This was a secondary analysis of the CADEUS cohort study on 5651 actively employed patients, who submitted at least one claim for the reimbursement of a NSAID dispensation for a MSD between August 2003 and July 2004, in the French National Healthcare Insurance database. Questionnaires were sent to prescribing physicians to obtain diagnoses and the medical history, and to patients for their occupation, height and weight and smoking status. Multivariate logistic regression was used to study the determinants of a heavy use of NSAIDs defined as 'over four dispensations in one year with less than two months between any two'. RESULTS: Factors associated with heavy use of NSAIDs were age (Odds ratio (OR): 1.8 (ten years), 95% confidence interval (CI): 1.6 1.9), osteoarthritis (versus back pain) (OR: 1.8, 95% CI: 1.5-2.1), body mass index (superior to 30) (OR: 1.8, 95% CI: 1.5-2.2), and occupation (blue collar versus white collar workers) (OR: 1.4, 95% CI: 1.2-1.6). Blue collar workers also had a 20% higher prevalence of 5-year history of dyspepsia. No difference was observed between sexes or in the use of COX-2 selective inhibitors between occupations. CONCLUSION: Factors associated with occupational constraints that contribute to the severity of MSDs, may explain the heavier use of NSAIDs among blue collar workers in spite of a concurrent and past medical history of adverse reactions to this type of medication. PMID- 19133062 TI - Randomized clinical comparisons of diclofenac concentration in the soft tissues and blood plasma between topical and oral applications. AB - AIMS: To compare tissue concentrations of diclofenac resulting from topical and oral applications of diclofenac according to clinically recommended prescriptions. METHODS: Diclofenac sodium was applied to 14 subjects (four male and 10 female), who were scheduled to undergo knee arthroplasty due to osteoarthritis, according to the oral or topical prescription (a capsule containing 37.5 mg diclofenac sodium or two tapes containing a total of 30 mg diclofenac sodium). At 12 h after prescription, the diclofenac concentration in the fat, muscle and synovial tissues was measured with liquid chromatography and mass analysis. RESULTS: The diclofenac concentration in the muscle was significantly higher (P = 0.0196) after topical application (average 9.29 ng ml( 1)) than after oral application (0.66 ng ml(-1)), whereas there was no significant difference in the plasma diclofenac level (4.70 and 6.63 ng ml(-1)) between the two applications. The concentration in the synovial membrane was significantly (P = 0.0181) lower in the topical application (4.99 ng ml(-1)) than in the oral application (15.07 ng ml(-1)). CONCLUSIONS: Whereas plasma levels resulting from topical and oral applications of diclofenac according to clinically recommended prescriptions were comparable, concentration levels in the muscle and synovial tissues were different. PMID- 19133066 TI - Terminology of the BSCC, European Community and the Bethesda system: the boundary between low-grade and high-grade cytology. PMID- 19133067 TI - European guidelines for clinical management of abnormal cervical cytology, part 2. AB - The current paper presents the second part of chapter 6 of the second edition of the European Guidelines for Quality Assurance in Cervical Cancer Screening. The first part of the same chapter was published in a previous issue (Cytopathology 2008;19:342-54). This part provides guidance on how to manage and treat women with histologically confirmed cervical intraepithelial neoplasia. The paper describes the characteristics, indications and possible complications of excisional and ablative treatment methods. The three options to monitor the outcome after treatment (repeat cytology, HPV testing and colposcopy) are discussed. Specific recommendations for particular clinical situations are provided: pregnancy, immuno-suppression, HIV infection, post-menopause, adolescence and cyto-colpo-histological disparity. The paper ends with recommendations for quality assurance in patient management and some general advice on how to communicate screening, diagnosis and treatment results to the woman concerned. Finally, a data collection form is attached. PMID- 19133068 TI - From the chief editor. PMID- 19133070 TI - Saccharomyces cerevisiae: the effect of different forms and concentrations of iodine on uptake and yeast growth. AB - The essentiality of iodine for humans, especially in the early stages of life, is well recognized. The chemical forms of iodine in food supplements, infant formulae and iodated salt are either iodide (KI) or iodate (KIO(3)). Because there are no or rare data about iodine uptake by yeasts, we investigated the influence of different sources of iodine, as KI, KIO(3) and periodate (KIO(4)), on its uptake in and growth of the model yeast Saccharomyces cerevisiae. KIO(3) inhibited the growth of the yeast the most and already at a 400 microM initial concentration in the growth medium; the OD was reduced by 23% in comparison with the control, where no KIO(3) was added. The uptake of different iodine sources by the yeast S. cerevisiae was minimal, in total <1%. Tracer experiments with radioactive (131)I added as KI showed that the yeast S. cerevisiae does not have the ability to transform KI into volatile species. We investigated the specificity of iodine uptake added as KIO(3) in the presence of Na(2)SeO(4) or ZnCl(2) or K(2)CrO(4) in the growth medium, and it was found that chromate had the most influence on reduction of KIO(3) uptake. PMID- 19133069 TI - In silico evidence for functional specialization after genome duplication in yeast. AB - A fairly recent whole-genome duplication (WGD) event in yeast enables the effects of gene duplication and subsequent functional divergence to be characterized. We examined 15 ohnolog pairs (i.e. paralogs from a WGD) out of c. 500 Saccharomyces cerevisiae ohnolog pairs that have persisted over an estimated 100 million years of evolution. These 15 pairs were chosen for their high levels of asymmetry, i.e. within the pair, one ohnolog had evolved much faster than the other. Sequence comparisons of the 15 pairs revealed that the faster evolving duplicated genes typically appear to have experienced partially--but not fully--relaxed negative selection as evidenced by an average nonsynonymous/synonymous substitution ratio (dN/dS(avg)=0.44) that is higher than the slow-evolving genes' ratio (dN/dS(avg)=0.14) but still <1. Increased number of insertions and deletions in the fast-evolving genes also indicated loosened structural constraints. Sequence and structural comparisons indicated that a subset of these pairs had significant differences in their catalytically important residues and active or cofactor binding sites. A literature survey revealed that several of the fast-evolving genes have gained a specialized function. Our results indicate that subfunctionalization and even neofunctionalization has occurred along with degenerative evolution, in which unneeded functions were destroyed by mutations. PMID- 19133071 TI - Concordant gene regulation related to perturbations of three GDP-mannose-related genes. AB - Glycosylation of proteins is one of the most crucial post-translational modifications. In order to access system-level and state-dependent data related to the regulation of glycosylation events, we cultivated yeast cell strains each harboring a selected conditional knockdown construct for a gene (either SEC53, VRG4 or DPM1) related to GDP-mannose synthesis or its utilization in glycan biosynthesis. In order to carry this out efficiently, we developed automated sampling from bioreactor cultivations, a collection of in silico workflows for data analysis as well as their integration into a large data warehouse. Using the above-mentioned approaches, we could show that conditional knocking down of transcripts related to GDP-mannose synthesis or transportation led to altered levels of over 300 transcripts. These transcripts and their corresponding proteins were characterized by their gene ontology (GO) annotations, and their putative transcriptional regulation was analyzed. Furthermore, novel pathways were generated indicating interactions between GO categories with common proteins, putative transcriptional regulators of such induced GO categories, and the large protein-protein interaction network among the proteins whose transcripts indicated altered expression levels. When these results are always added to an ever-expanding data warehouse as annotations, they will incrementally increase the knowledge of biological systems. PMID- 19133072 TI - Vanderwaltozyma verrucispora sp. nov., a new ascomycetous yeast species. AB - A new yeast species, Vanderwaltozyma verrucispora, is proposed in this study based on two strains isolated from partially decayed leaves in Japan and one strain from soil in Taiwan. The species is characterized by the fermentation of glucose and galactose, formation of one to four spheroidal to ellipsoidal ascospores with warty surfaces in each ascus, and assimilation of a few carbon and nitrogen compounds. Genus assignment and distinction of the species from the other two recognized species of Vanderwaltozyma is based on the morphological and physiological characteristics, and phylogenetic analysis of nucleotide sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene. From these comparisons, the name V. verrucispora sp. nov. is proposed. Sequence analysis of the D1/D2 domains of the LSU rRNA gene reveals that the phylogenetically closest relative of V. verrucispora is Vanderwaltozyma yarrowii. The type strain of the new species, which was isolated from a partially decayed leaf in Kagoshima Prefecture, Japan, is NBRC 1884(T) (=CBS 10887(T)=BCRC 23141(T)). PMID- 19133073 TI - Mining an Ostrinia nubilalis midgut expressed sequence tag (EST) library for candidate genes and single nucleotide polymorphisms (SNPs). AB - Genes expressed in lepidopteran midgut tissues are involved in digestion and Bacillus thuringiensis (Bt) toxin resistance traits. Five hundred and thirty five unique transcripts were annotated from 1745 high quality O. nubilalis larval midgut expressed sequence tags (ESTs). Full-length cDNA sequence of 12 putative serine proteinase genes and 3 partial O. nubilalis aminopeptidase N protein genes, apn1, apn3, and apn4, were obtained, and genes may have roles in plant feeding and Bt toxin resistance traits of Ostrinia larvae. The EST library was not normalized and insert frequencies reflect transcript levels under the initial treatment conditions and redundancy of inserts from highly expressed transcripts allowed prediction of putative single nucleotide polymorphisms (SNPs). Ten di-, tri- or tetranucleotide repeat unit microsatellite loci were identified, and minisatellite repeats were observed within the C-termini of two encoded serine proteinases. Molecular markers showed polymorphism at 28 SNP loci and one microsatellite locus, and Mendelian inheritance indicated that markers were applicable to genome mapping applications. This O. nubilalis larval midgut EST collection is a resource for gene discovery, expression information, and allelic variation for use in genetic marker development. PMID- 19133074 TI - The gustatory receptor family in the silkworm moth Bombyx mori is characterized by a large expansion of a single lineage of putative bitter receptors. AB - The gustatory receptor (Gr) family of insect chemoreceptors includes receptors for sugars and bitter compounds, as well as cuticular hydrocarbons and odorants such as carbon dioxide. We have annotated a total of 65 Gr genes from the silkworm Bombyx mori genome. The Gr family in the silkworm moth includes putative carbon dioxide receptors and sugar receptors, as well as duplicated orthologues of the orphan DmGr43a receptor. Most prominent in this 65-gene family, however, is a single large expansion of 55 Grs that we propose are predominantly 'bitter' receptors involved in perception of the large variety of secondary plant chemicals that caterpillars and moths encounter. These Grs might therefore mediate food choice and avoidance as well as oviposition site preference. PMID- 19133075 TI - Genome-wide analysis of genes related to ovary activation in worker honey bees. AB - A defining characteristic of eusocial animals is their division of labour into reproductive and nonreproductive specialists. Here, we used a microarray study to identify genes associated with functional sterility in the worker honey bee Apis mellifera. We contrasted gene expression in workers from a functionally sterile wild-type strain with that in a mutant (anarchist) strain selected for high rates of ovary activation. We identified a small set of genes from the brain (n = 7) and from the abdomen (n = 5) that are correlated in their expression with early stages of ovary activation. Sterile wild-type workers up-regulated two unknown genes and a homologue of Drosophila CG6004. By contrast, reproductive anarchist workers up-regulated genes for the yolk protein vitellogenin, venom peptides and a member of the AdoHycase superfamily, among others. The differentially expressed genes identified are likely to be involved in early differentiation into sterile and reproductive worker phenotypes and may therefore form part of the gene networks associated with the regulation of honey bee worker sterility. Our study may have lacked sufficient power to detect all but a minority of biologically relevant changes taking place; however, the differential expression of vitellogenin and a putative AdoHycase suggests that our screen has captured core reproductive genes and that ovary activation may involve an epigenetic mechanism. PMID- 19133076 TI - Isolation and functional characterization of two independently-evolved fatty acid Delta12-desaturase genes from insects. AB - We report the first isolation and characterization of insect fatty acid Delta12 desaturase genes, AdD12Des from house cricket (Acheta domesticus) and TcD12Des from the red flour beetle (Tribolium castaneum), responsible for the production of linoleic acid from oleic acid. Sequence analysis shows the cricket and flour beetle Delta12-desaturase genes have evolved independently from all previously known Delta12-desaturases and are much more closely related to the archetypal stearoyl-Coenzyme A-acting desaturase from rat than to the phospholipid-acting Delta12-desaturases widely reported in plants. Phylogenetic and functional analysis indicates the cricket AdD12Des gene may have evolved from an ancestral Delta9-desaturase. By contrast, the beetle Delta12-desaturase is distantly related to the cricket genes and beetle Delta9-desaturases suggesting evolution by an independent route. Linoleic acid has key physiological roles in insects and this is the first report of genes capable of producing this essential fatty acid in higher animals. PMID- 19133077 TI - Chikungunya-Wolbachia interplay in Aedes albopictus. AB - A severe Chikungunya (CHIK) outbreak recently hit several countries of the Indian Ocean. On La Reunion Island, Aedes albopictus was incriminated as the major vector. This mosquito species is naturally co-infected with two distinct strains of the endosymbiont Wolbachia, namely wAlbA and wAlbB, which are increasingly attracting interest as potential tools for vector control. A PCR quantitative assay was developed to investigate Wolbachia/mosquito host interactions. We show that Wolbachia densities are slightly decreased in CHIK virus (CHIKV)-infected females. We measured the impact of CHIKV replication on a lysogenic virus: WO bacteriophage. Our data indicate that WO is sheltered by wAlbB, likely at a single copy per bacteria, and that CHIKV replication is not a physiological stress triggering WO entrance into the lytic cycle. PMID- 19133078 TI - Nucleotide diversity in the Hsp90 gene in natural populations of Drosophila melanogaster from Australia. AB - Hsp90 is regarded as one of the best candidates for an evolved mechanism that regulates the expression of genetic and phenotypic variability. We examined nucleotide diversity in both the promoter and coding regions of Hsp90, the gene which encodes Hsp90 in Drosophila, in natural populations of Drosophila melanogaster from eastern Australia. We found that Hsp90 is polymorphic for only two nonsynonymous changes in the coding region, both of which are deletions of a lysine residue. One of these lysine deletions was in complete linkage disequilibrium with the inversion In(3L)P, and showed a significant association with latitude. The other lysine deletion reported here for the first time varied from 0 to 15% in natural populations, but did not show a clinal pattern. The regulatory and coding regions of Hsp90 showed very low nucleotide diversity compared to other nuclear genes, and chromosomes containing In(3L)P had lower levels of nucleotide diversity than the standard arrangements. Non-neutral evolution of Hsp90 was not supported by analyses of either the regulatory or coding regions of the gene. These results are discussed within the context of Hsp90 variation being involved in thermotolerance as well as the expression of genetic and phenotypic variability. PMID- 19133079 TI - The hsp27 gene of the Mediterranean fruit fly, Ceratitis capitata: structural characterization, regulation and developmental expression. AB - In the present study, a genomic DNA clone encoding the medfly homolog of Drosophila melanogaster hsp27 gene, named Cchsp27, was isolated. We sequenced a part of the clone containing the coding region, the 5' untranslated region and approximately 2.8 Kb of the 5' flanking region of the gene. Phylogenetic analysis of several insect small heat shock proteins, suggested that CcHsp27 is orthologous to Drosophila Hsp27 and Sarcophaga crassipalpis Hsp25. The Cchsp27 gene was mapped at the 81A division of the sixth chromosome which coincides with one of the major heat shock puffs of medfly. Structural analysis of the 5' flanking region of the Cchsp27 gene revealed the presence of five putative heat shock elements and one putative ecdysone response element. In addition to heat induction, the Cchsp27 gene was expressed at several stages of normal medfly development. In general, the developmental expression pattern of the Cchsp27 gene was similar to the respective pattern of Drosophila hsp27 gene. However, there were some important differences in certain developmental stages suggesting differential regulation of the hsp27 gene in the two dipterans species. Salivary gland culture experiments showed that the Cchsp27 gene is regulated by 20 hydroxyecdysone. PMID- 19133080 TI - Professor Thomas Pitt Ford. PMID- 19133081 TI - Frequency of radiological review in pulp studies. PMID- 19133082 TI - Calcium ion release from calcium hydroxide stimulated fibronectin gene expression in dental pulp cells and the differentiation of dental pulp cells to mineralized tissue forming cells by fibronectin. AB - AIM: The effect of calcium ions on dental pulp cells was examined and the mechanism of dentine bridge formation by calcium hydroxide was investigated. METHODOLOGY: Human dental pulp cells were treated with high concentration of calcium or magnesium ions for 24 h and fibronectin gene expression was measured by the quantitative PCR method. Human dental pulp cells were then cultured on fibronecin-coated dishes for 24 h, and osteocalcin and osteopontin gene expression, which are typical phenotypes of mineralized tissue forming cells, were measured by the quantitative PCR method. RESULTS: Fibronectin gene expression was stimulated by calcium ions dose-dependently. On the other hand, magnesium ions did not influence fibronectin gene expression. Furthermore, pulp cells cultured on fibronectin-coated dishes enhanced the expression of phenotypes of mineralized tissue forming cells. CONCLUSIONS: Calcium ions released from calcium hydroxide stimulates fibronectin synthesis in dental pulp cells. Fibronectin might induce the differentiation of dental pulp cells to mineralized tissue forming cells that are the main cells to form dentine bridges, via contact with cells. PMID- 19133083 TI - Mechanical behaviour of nickel-titanium rotary endodontic instruments in simulated clinical conditions: a computational study. AB - AIM: To develop an accurate finite element (FE) model for studying rotary endodontic instruments and to demonstrate the usefulness of the FE method in improving the knowledge of the mechanical behaviour of these instruments during root canal preparation. METHODOLOGY: An accurate geometrical model of a Ni-Ti ProTaper F1 instrument was created. The interaction between the rotating instrument and differently shaped root canals during the insertion and removal procedure was studied using FE analyses. The complex thermo-mechanical behaviour of the Ni-Ti alloy was reproduced using an ad hoc computational subroutine. With the aim of demonstrating the enhanced performance of the shape memory alloy employment, the same analysis was performed on a 'virtual' ProTaper F1 made of stainless steel. RESULTS: The Ni-Ti instrument operated in its pseudo-elastic range and was able to recover its original shape and to follow the canal curvature without deviation. The radius and the position of the canal curvature are the most critical parameters that determined the stress in the instrument with higher stress levels being produced by decreasing the radius and moving from the apical to the mid root position. CONCLUSIONS: The most demanding working conditions were observed in canals with sharp curves, especially in areas where the instruments had larger diameters. To prevent possible damage to instruments and fracture, it is advised that the instruments should be discarded following their use in such canals. PMID- 19133084 TI - Ex vivo evaluation of dye penetration associated with various dentine bonding agents in conjunction with different irrigation solutions used within the pulp chamber. AB - AIM: To compare the effect of sodium hypochlorite (NaOCl), chlorhexidine, povidone iodine and sodium ascorbate on dye penetration associated with various dentine adhesives used within the pulp chamber. METHODOLOGY: One hundred and sixteen mandibular molar teeth were divided into eight groups of 12 teeth each by irrigation regimen and adhesive system. The roof of pulp chambers and roots were removed under water cooling. The pulp chambers in the experimental groups were irrigated with 5.25% NaOCl, 0.2% chlorhexidine, 5% w/v povidone iodine or 5.25% NaOCl followed by sodium ascorbate and restored with Surefil using either Prime & Bond NT or Xeno III with each irrigation regimen. Twenty teeth were used as control specimens in which no disinfectant was used before bonding. Ten teeth from each group were immersed in 2% methylene blue dye and assessed for dye penetration. The data were statistically analysed by Kruskal-Wallis and Mann Whitney U-tests. Two samples in each group were observed under scanning electron microscopy for interfacial gap evaluation. RESULTS: No significant difference was found in dye leakage between control and groups with 5.25% NaOCl pretreatment with both adhesive systems. Chlorhexidine and povidone iodine pretreatment resulted in significantly less dye penetration with Xeno III as compared with Prime & Bond NT. Sodium ascorbate treatment following NaOCl application significantly reduced microleakage and improved the marginal adaptation with both adhesive systems. CONCLUSIONS: The type of irrigant affected the sealing ability of bonding agents inside the pulp chamber and was material specific. Xeno III had less dye penetration with CHX and povidone iodine whilst sodium ascorbate pretreatment after NaOCl reduced dye penetration with both PBNT and Xeno III. PMID- 19133085 TI - Technical quality of root fillings and periapical status in root filled teeth in Jonkoping, Sweden. AB - AIM: To study changes in technical quality of root fillings and periapical status in root filled teeth in random samples of 20 to 70 year-olds between 1973 and 2003. METHODOLOGY: Data from dentate subjects aged 20-70 years from examinations conducted in 1973 (n = 498), 1983 (n = 530), 1993 (n = 547) and 2003 (n = 491) were used for the analysis. Length of root fillings were measured on radiographs to the nearest 0.1 mm. Adequate seal was defined as a root filling without lateral and/or apical voids. Periapical status was assessed according to the periapical index. All observations were made by one calibrated observer registered from full mouth radiographic examinations. Teeth with root fillings ending within the canal without lateral or apical voids were considered adequate. The association between root filling quality and periapical status was analysed by means of the Chi-squared test and a multilevel logistic regression analysis. RESULTS: Adequately root filled teeth had a significantly lower frequency of apical periodontitis than inadequately root filled teeth (11.8% vs. 22.8%). The frequency of technically adequate root fillings increased statistically significant from 1973 (23.7%) to 2003 (36.4%) without a concomitant improvement of the periapical status in root filled teeth over time (24.5% vs. 24.6%). CONCLUSIONS: This study reports on an improving technical quality of root fillings over time, without a concomitant improvement of the periapical status in root filled teeth. A larger proportion of treated molars over time may be of importance for the result. PMID- 19133086 TI - Effect of remaining dentine structure and thermal-mechanical aging on the fracture resistance of bovine roots with different post and core systems. AB - AIM: To evaluate the influence of remaining dentine thickness around post and core systems and the thermo-mechanical stresses on fracture resistance of bovine roots. METHODOLOGY: This study involved 288 bovine incisor roots with standardized dimensions. Roots were randomly distributed into 24 groups (n = 12) according to root conditions [intact, semi-weakened, or weakened] and post and core systems [custom cast core, composite resin core, prefabricated metallic post, or prefabricated carbon fibre post], submitted or not to thermomechanical aging [5000 thermal cycles and 100 000 mechanical cycles at a 135-degree angle to the long axis of the root]. Specimens were submitted to a tangential compressive load (135 degrees angle) at a crosshead speed of 0.5 mm min(-1) until failure. Fracture resistance data were analyzed using 3-way anova and Tukey test: alpha = 5%. RESULTS: Roots restored with composite resin cores demonstrated no resistance to mechanical aging. No statistically significant difference was observed between aged and nonaged specimens involving all post-systems. Roots restored with custom cast cores had the highest fracture strength, followed by prefabricated metallic posts and carbon fibre posts, regardless of root conditions and thermomechanical aging. The remaining dentine thickness affected significantly roots restored with custom cast cores; weakened roots had a lower fracture resistance. CONCLUSIONS: Although custom cast cores had a higher fracture resistance when compared to the other techniques, the results were highly dependent on remaining dentine thickness. Prefabricated posts performed in a similar manner in intact, semi weakened and weakened roots reinforced with composite resin. PMID- 19133087 TI - Microscopic appearance and apical seal of root canals filled with gutta-percha and ProRoot Endo Sealer after immersion in a phosphate-containing fluid. AB - AIM: To investigate the sealing quality of ProRoot Endo Sealer, a calcium silicate-based sealer and its morphologic characteristics after immersion in a phosphate-containing fluid (PCF). METHODOLOGY: Single-rooted canals were filled with gutta-percha and either ProRoot Endo Sealer or two commercially available zinc oxide eugenol (ZOE)-based and epoxy resin-based sealers. The sealers were allowed to set for 6 days and the filled teeth were immersed in PCF for 24 h before fluid leakage evaluation. After initial leakage evaluation at the 7th day, each filled root was restored and reimmersed in PCF for 28 days before the second phase of leakage evaluation at 35 days. Cryofractured specimens of additional teeth filled with the three sealers were examined using scanning electron microscopy after immersion in PCF for the two periods. RESULTS: One-way repeated measures anova and Tukey test revealed significant differences between the ZOE based sealer at 35 days and the calcium silicate-based sealer at 35 days (P < 0.001), and between the ZOE-based sealer at 7 days and the calcium silicate-based sealer at 35 days (P = 0.001). No difference was found between the epoxy resin based sealer and the calcium silicate-based sealer after both storage periods. Cryofractured calcium silicate-based sealer specimens demonstrated apatite-like crystalline deposits along the apical and middle thirds of the canal walls via transformation from amorphous calcium phosphate-like precursors. CONCLUSIONS: ProRoot Endo Sealer is comparable in sealing quality to the epoxy resin-based sealer and seals better than the ZOE-based sealer after immersion in PCF. The calcium silicate-based sealer also demonstrates ex vivo bioactivity when it comes into contact with phosphate ions. PMID- 19133088 TI - Assessment of vascularity as an index of angiogenesis in periradicular granulomas. Comparison with oral carcinomas and normal tissue counterparts. AB - AIM: To quantify vascularity in periradicular granulomas using different endothelial markers, and assess its value as an index of angiogenesis by comparing granulomas with healthy periodontal ligament (PDL). To use oral tumours, compared with adjacent normal mucosa, as positive controls. METHODOLOGY: Paraffin-embedded sections were stained with antibodies to von Willebrand factor (vWF), a pan-endothelial marker, and CD105, a putative marker for angiogenic vessels. Vascularity was quantified by different methods reflecting vessel volume and density. RESULTS: Irrespective of the marker or method used, vascularity values were similar in periradicular granuloma and PDL. Both tissues were highly vascularized, with levels similar to those found in oral squamous cell carcinoma. Vascularity was significantly higher in the latter than in normal mucosa. Fewer vessels were positive for CD105 than for vWF in the normal mucosa, whereas similar numbers were found in the other tissues examined. CONCLUSIONS: A comparison of vascularity in oral tumours and normal oral mucosa provided evidence of angiogenesis in the former. Staining with CD105 added limited value to staining with vWF in these tissues. In contrast, a comparison of periradicular granuloma and PDL failed to demonstrate evidence of angiogenesis in the granuloma. As all vessels were similarly stained with vWF and CD105 in granuloma and PDL, a possible hypothesis is that all vessels are newly formed in these tissues. A more plausible alternative is that CD105 expression may reflect the metabolic activity or intrinsic characteristics of the tissues, rather than the presence of angiogenic vessels. PMID- 19133089 TI - External apical root resorption: two case reports. AB - AIM: To present two cases of external root resorption and in so doing highlight rarer causative factors as well as the difficulties in classifying root resorption. SUMMARY: Root resorption in the permanent dentition is usually pathological. Local factors are the most frequent causes of resorption, most commonly excessive pressure and inflammation. However, many systemic abnormalities have been implicated in the resorption process. Resorption is often an incidental finding during routine examination. Otherwise, late in the disease process, tooth mobility or infection may arise. A rarer form of root resorption is external apical root resorption which may or may not be linked to systemic disease. In most cases, no causative factors are found. At present there is no curative treatment for external apical root resorption. The current management for these patients is symptomatic, minimal intervention and long-term monitoring. KEY LEARNING POINTS: * Apical resorption may be associated with a systemic disease or of an idiopathic origin. It may also occur in association with orthodontic treatment or with pathoses such as tumours, cysts, etc. * In the absence of signs or symptoms of pulpal and/or periapical disease, endodontic treatment is not indicated. * Long-term monitoring of affected patients is essential. PMID- 19133090 TI - Anomalous mandibular premolars: a mandibular first premolar with three roots and a mandibular second premolar with a C-shaped canal system. AB - AIM: To describe unusual variations in the root morphology and root canal systems of mandibular first and second premolar teeth extracted for orthodontic reasons. SUMMARY: Normally mandibular first and second premolar teeth have single roots with single canals. A 15-year-old patient presented for orthodontic treatment and two mandibular premolar teeth were examined post-extraction. The mandibular first premolar exhibited three distinct, separate roots and the mandibular second premolar exhibited a C-shaped root canal system. The coronal morphology of each of the mandibular premolars revealed dimensions and anatomy within normal limits. The incidence of a three-rooted mandibular first premolar is approximately 0.2%. KEY LEARNING POINTS: * Thorough clinical and radiographic interpretation is important in recognizing anomalous root and root canal systems. * The most common forms of root and canal systems and its aberrations must be understood to realize variations from normal do occur. * Successful root canal treatment requires an accurate diagnosis of the root canal system using all available aids. * Value of microcomputed tomography in the study of anatomy ex vivo and cone-beam tomography in clinical endodontics of complex premolar cases is increasing. PMID- 19133091 TI - Step-wise treatment of two periodontal-endodontic lesions in a heavy smoker. AB - AIM: To report a clinical case of two advanced periodontal-endodontic lesions with a focus on treatment issues related to tobacco use. SUMMARY: A 53-year-old Caucasian male was referred to the School of Dentistry, Basel, Switzerland, for periodontal treatment. The major diagnoses were chronic (smoker) periodontitis and advanced combined periodontal-endodontic lesions on the mandibular left lateral incisor and right incisor. Conventional root canal treatment was performed, and subsequently led to reduced radiolucencies around the affected roots after 14 months. The remaining osseous defect was augmented by guided tissue regeneration using bovine bone substitute and resorbable membrane. The follow-up revealed a stable situation from clinical (probing depth 2-4 mm) and radiological points of view 32 months after initiation of treatment. Treatment considerations related to tobacco use are discussed. KEY LEARNING POINTS: * After conventional root canal treatment, osseous healing should occur before further complementary therapy is taken into account. * Issues related to tobacco use have to be considered before treatment is initiated. PMID- 19133092 TI - The International Endodontic Journal--a contemporary publication. PMID- 19133093 TI - Outcome of secondary root canal treatment: a systematic review of the literature. AB - AIMS (I): To investigate the effects of study characteristics on the reported success rates of secondary root canal treatment (2 degrees RCT or root canal retreatment); and (ii) to investigate the effects of clinical factors on the success of 2 degrees RCT. METHODOLOGY: Longitudinal human clinical studies investigating outcome of 2 degrees RCT which were published upto the end of 2006 were identified electronically (MEDLINE and Cochrane database 1966-2006 Dec, week 4). Four journals (Dental Traumatology, International Endodontic Journal, Journal of Endodontics, Oral Surgery Oral Medicine Oral Pathology Endodontics Radiology), bibliographies of all relevant papers and review articles were hand-searched. Two reviewers (Y-LN, KG) independently assessed and selected the studies based on specified inclusion criteria and extracted the data onto a pre-designed proforma, independently. The criteria were: (i) Clinical studies on 2 degrees RCT; (ii) Stratified analyses available for 2 degrees RCT where 1 degrees RCT data included; (iii) Sample size given and larger than 10; (iv) At least 6-month post operative review; (v) Success based on clinical and/or radiographic criteria (strict = absence of apical radiolucency; loose = reduction in size of radiolucency); and (vi) Overall success rate given or could be calculated from the raw data. Three strands of evidence or analyses were used to triangulate a consensus view. The reported findings from individual studies, including those excluded for quantitative analysis, were utilized for the intuitive synthesis which constituted the first strand of evidence. Secondly, the pooled weighted success rates by each study characteristic and potential prognostic factor were estimated using the random effect model. Thirdly, the effects of study characteristics and prognostic factors (expressed as odds ratios) on success rates were estimated using fixed and random effects meta-analysis with DerSimonean and Laird's methods. Meta-regression models were used to explore potential sources of statistical heterogeneity. Study characteristics considered in the meta-regression analyses were: decade of publication, study-specific criteria for success (radiographic, combined radiographic & clinical), unit of outcome measure (tooth, root), duration after treatment when assessing success ('at least 4 years' or '<4 years'), geographic location of the study (North American, Scandinavian, other countries), and qualification of the operator (undergraduate students, postgraduate students, general dental practitioners, specialist or mixed group). RESULTS: Of the 40 papers identified, 17 studies published between 1961 and 2005 were included; none were published in 2006. The majority of studies were retrospective (n = 12) and only five prospective. The pooled weighted success rate of 2 degrees RCT judged by complete healing was 76.7% (95% CI 73.6%, 89.6%) and by incomplete healing, 77.2% (95% CI 61.1%, 88.1%). The success rates by 'decade of publication' and 'geographic location of study' were not significantly different at the 5% level. Eighteen clinical factors had been investigated in various combinations in previous studies. The most frequently and thoroughly investigated were 'periapical status' (n = 13), 'size of lesion' (n = 7), and 'apical extent of RF' (n = 5) which were found to be significant prognostic factors. The effect of different aspects of primary treatment history and re-treatment procedures has been poorly tested. CONCLUSIONS: The pooled estimated success rate of secondary root canal treatment was 77%. The presence of pre-operative periapical lesion, apical extent of root filling and quality of coronal restoration proved significant prognostic factors with concurrence between all three strands of evidence whilst the effects of 1 degrees RCT history and 2 degrees RCT protocol have been poorly investigated. PMID- 19133094 TI - Ex vivo antimicrobial activity of several bleaching agents used during the walking bleach technique. AB - AIM: To investigate ex vivo the antimicrobial activity of a paste of sodium perborate associated with various vehicles comparing it with 37% carbamide peroxide and 35% hydrogen peroxide. METHODOLOGY: The antimicrobial activity of these agents was evaluated against three microorganisms: Enterococcus faecalis, Streptococcus mutans and Candida albicans. One millilitre of each tested substance was placed on the bottom of wells of 24-well cell culture plates. Six wells were used for each time period and group. Two millilitres of the microbial suspension was ultrasonically mixed for 10 s with the bleaching pastes and placed in contact with them for 10, 30, 45 s; 1, 3, 5, 10, 20, 30 min; and 1 and 2 h. After each period of time, 1 mL from each well was transferred to tubes containing 2 mL of freshly prepared brain heart infusion agar + neutralizers. Agar plates were inoculated in appropriate gaseous conditions. Data were analysed statistically by the Kruskal-Wallis test with the level of significance set at P < 0.05. RESULTS: In all groups containing chlorhexidine (groups 3, 5 and 7), the antimicrobial activity of the bleaching paste was significantly increased when compared with groups with other kinds of vehicle (groups 1, 2, 4, 6 and 8). For all tested groups, the most resistant microorganism was E. faecalis. CONCLUSIONS: Chlorhexidine when used as a vehicle for sodium perborate enhanced its antimicrobial activity. PMID- 19133095 TI - Apical extrusion of intracanal bacteria following use of various instrumentation techniques. AB - AIM: To evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using a manual technique and three engine-driven techniques utilizing nickel-titanium instruments (K3, RaCe, and FlexMaster). METHODOLOGY: Seventy extracted human mandibular premolar teeth with similar dimensions were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and then dried. The contaminated roots were divided into four experimental groups of 15 teeth each and one control group of 10 teeth. G1. RaCe group: the root canals were instrumented using RaCe instruments. G2. K3 group: the root canals were instrumented using K3 instruments. G3. FlexMaster group: the root canals were instrumented using FlexMaster instruments. G4. Manual technique group: the root canals were instrumented using K-type stainless steel instruments. G5. CONTROL GROUP: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The resultant microbiological samples were removed from the vials and then incubated in culture media for 24 h. The number of colony-forming units (CFU) was determined for each sample. The data obtained were analysed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. RESULTS: There was a significant difference between experimental-control and engine-driven-manual technique groups (P < 0.05). The manual technique was associated with the greatest extrusion of microorganism. CONCLUSIONS: All instrumentation techniques extruded intracanal bacteria apically. No significant difference was found in the number of CFU among the engine-driven techniques; manual techniques extruded significantly more microorganisms. PMID- 19133097 TI - Opinions and attitudes of endodontists and general dental practitioners in the UK towards the intra-canal fracture of endodontic instruments. Part 2. AB - AIM: To investigate the attitudes of general dental practitioners (GDPs) and endodontists in the UK towards management of fractured endodontic instruments. METHODOLOGY: A questionnaire was sent to 330 systemically selected GDPs and all endodontists working in the UK (170). It was accompanied by a covering letter explaining the aims of the study and indicating that all the information given would remain confidential. Those who did not respond to the first mailing were sent another two mailings. Data were analysed using chi-square test at P 70% cytotoxicity). CONCLUSIONS: The current results suggest that some endodontic materials have an elevated biological risk for extended intervals. PMID- 19133098 TI - Efficacy of rotary instruments with greater taper in preparing oval root canals. AB - AIM: To compare the preparation quality of two rotary systems and NiTi-hand files in oval root canals, and to evaluate the effect of canal dimensions on the preparation. METHODOLOGY: Ninety roots with oval root canals were selected. The middle third was cross sectioned at two levels and photographed. The maximum and minimum diameters of the root and canal were recorded. Teeth were distributed in three groups (n = 30) using stratified randomization, and prepared under simulated clinical conditions with Mtwo, ProTaper, or NiTi-hand files. The pre- and post-preparation photographs were traced and superimposed, the thickness of dentine removed was measured and the ratio of prepared canal outline was calculated. The impact of preparation system and canal dimensions on the quality of the preparation was evaluated using regression analysis. RESULTS: With regards to the ratio of prepared canal outline, no statistical significant difference was found between Mtwo (0.75 [95%CI: 0.69; 0.81]) and ProTaper (0.75 [95%CI: 0.69; 0.80]), but both systems performed significantly better than NiTi-hand files (0.65 [95%CI: 0.60; 0.71]). In six root canals in Mtwo-group (20%), and eight root canals in ProTaper-group (27%), the minimal thickness of dentine-wall after preparation was less than 0.5 mm. In contrast to the maximum diameter of the root canal, the minimum diameter influenced the quality of the preparation (P = 0.0006). CONCLUSIONS: No instrumentation technique was able to circumferentially prepare the oval outline of root canals. Nevertheless, instruments with greater taper (ProTaper and Mtwo) were more efficient than NiTi- hand files, but this was, in some cases, at the expense of remaining dentine-wall thickness. PMID- 19133099 TI - Degradation properties and ion release characteristics of Resilon and phosphate glass/polycaprolactone composites. AB - AIMS: To (i) characterise the degradation pattern and ion release characteristics of bioactive glass/polycaprolactone (PCL) composites; (ii) compare the degradation behaviour of composites containing phosphate glass to that of Resilon; and (iii) determine the potential to control the degradation of the composites by modifying the iron content of phosphate glasses. METHODOLOGY: The degradation behaviour of the phosphate glass/PCL samples (n = 3) and that of Resilon were determined in two aqueous solutions (buffered distilled water, Hanks Buffered Saline Solution) over a 7-day period using weight change measurement, ion chromatography, light microscopy (LM) and scanning electron microscopy (SEM). RESULTS: An initial increase in weight of the samples was followed by loss. All phosphate glass/PCL composites released various amounts of iron, sodium, calcium and phosphate ions into solution. The Resilon samples released only sodium and calcium ions. The visual methods (LM, SEM) showed distinct precipitate (calcium phosphate) formation on the surface of the phosphate glass/PCL composites but only a faint surface residue on the surface of Resilon. CONCLUSIONS: The degradation pattern of the composites containing phosphate glass was different from that of Resilon. The phosphate glass/PCL composites encouraged precipitate formation on the material surface, which may be advantageous in creating a marginal and apical seal. PMID- 19133100 TI - Effect of EDTA conditioning upon the retention of fibre posts luted with resin cements. AB - AIM: To determine if a final rinse with EDTA improves the short-term in-tooth retentive strength of adhesively luted fibre-reinforced composite posts. METHODOLOGY: Six different commercial fibre post systems were tested. Post spaces in extracted human anterior teeth were irrigated with either 6% NaOCl or 6% NaOCl followed by 17% EDTA. Posts were luted using the manufacturer's recommended bonding agent and luting cement. Two different sizes of post were tested for each post system. After 24 h of storage, the retentive strength of the post to the tooth (14 per group, 336 total) was tested using a pull-out technique. Three-way anova and the Student-Newman-Kuels test were used to test the effects of irrigation regimen, post size and post brand on the retentive strength of the fibre post. RESULTS: Almost every sample failed at the interface between dentine and luting cement. Irrigation method did not have any significant effect (P > 0.14) for any group other than ParaPost Fibre White (P < 0.001). The overall retention of a post appeared to be correlated to the amount of surface texture on the post; the smooth ICEPost was the least retentive brand, the textured FibreKor and the ParaPost Fibre White posts were moderately retained, and the threaded Flexi-Post Fibre and the Flexi-Flange Fibre posts were the most retentive brands tested. For all brands, larger diameter posts were more retentive than smaller diameter posts (P < 0.008). CONCLUSIONS: A final rinse of EDTA did not improve the short-term retention of fibre posts except for ParaPost Fibre White. PMID- 19133101 TI - The biocompatibility of modified experimental Portland cements with potential for use in dentistry. AB - AIM: To evaluate the biocompatibility of a group of new potential dental materials and their eluants by assessing cell viability. METHODOLOGY: Calcium sulpho-aluminate cement (CSA), calcium fluoro-aluminate cement (CFA) and glass ionomer cement (GIC; Ketac Molar), used as the control, were tested for biocompatibility. Using a direct test method cell viability was measured quantitatively using alamarBluetrade mark dye, and an indirect test method where cells were grown on material elutions and cell viability was assessed using methyltetrazolium (MTT) assay as recommended by ISO 10 993-Part 5 for in vitro testing. Statistical analysis was performed by analysis of variance and Tukey multi-comparison test method. RESULTS: Elution collected from the prototype cements and the GIC cured for 1 and 7 days allowed high cell activity after 24 h cell exposure, which reduced after 48 h when compared to the nontoxic glass ionomer control, but increased significantly after 72 h cell contact. Elutions collected after 28 days revealed reduced cell activity at all cell exposure times. Cells placed in direct contact with the prototype materials showed reduced cell activity when compared with the control. CONCLUSIONS: Cell growth was poor when seeded in direct contact with the prototype cements. GIC encouraged cell growth after 1 day of contact. The eluted species for all the cements tested exhibited adequate cell viability in the early ages with reduced cell activity at 28 days. Changes in the production of calcium hydroxide as a by-product of cement hydration affect the material biocompatibility adversely. PMID- 19133102 TI - Influence of tooth age and root section on root dentine dye penetration. AB - AIM: To investigate dye permeability of root dentine according to patients' age, root section and dye penetration time. METHODOLOGY: A total of 96 extracted human single-rooted teeth, assigned to four age groups (<30, 30-45, 45-60 and >60 years) were separated at the cemento-enamel junction and root canals were enlarged. The root surfaces were coated with cyanocrylate to prevent external dye penetration and centrifuged in distilled water to eliminate air. For dye penetration the root canals were filled with methylene blue 5%. After 1, 30 and 60 days eight roots per age group were cross-sectioned in 1 mm slices. Dye penetrated areas and the complete dentine areas were digitized and measured. Differences between groups were judged with anova and LSD, P < 0.05 or P < 0.01. RESULTS: The root section, the patients' age and the penetration time influenced significantly the penetrated areas (P < 0.05). After 1 and 30 days significant differences could be found only in the apical root sections between all age groups (P < 0.05). Dye penetration areas systematically decreased with increasing age and also from coronal to apical (P < 0.01). CONCLUSIONS: Age influenced dye penetration significantly. Dye penetration also depended on the location (coronal, middle and apical) within the root canal. These findings indicate that there may be a correlation between the tooth age and permeability of root dentine, which may influence the distribution and effectiveness of drugs used for root canal disinfection. PMID- 19133103 TI - Dens invaginatus. Part 1: classification, prevalence and aetiology. AB - AIM: To review and discuss the aetiology, prevalence and classification of this dental anomaly together with the morphology of an invagination and the most appropriate nomenclature. SUMMARY: This review considers the different possible nomenclatures and concludes that dens invaginatus is the most appropriate description. The paper highlights the different reported prevalence figures and concludes that the problem is probably one of the most common of the dental developmental abnormalities with maxillary lateral incisors most commonly affected. The paper suggests that the classification system described by Oehlers (1957a) is probably the most clinically relevant and that the morphological features associated with this problem may increase the risk of pulpal pathology developing and complicate any possible endodontic treatment. KEY LEARNING POINTS: * The aetiology of dens invaginatus is still unknown, although there is some evidence that it may be genetic in origin. * The problem is probably more prevalent than most clinicians are aware of and this is because of the diagnostic difficulties associated with the anomaly. * The nature of the problem may increase the risk of pulp disease and complicate any root canal treatment. PMID- 19133104 TI - Dens invaginatus. Part 2: clinical, radiographic features and management options. AB - AIM: To describe the possible clinical and radiographic features of this developmental anomaly, review previous treatment recommendations and suggest management options based on the classification of the problem. SUMMARY: This paper describes the clinical and radiographic features related to the different types of dens invaginatus and highlights those features which may indicate the presence of a previously undetected invagination. Aids to clinical diagnosis are described together with a description of the possible radiographic features, which may suggest the presence of an invagination. Previous treatment suggestions are described and suggestions as to possible management options, based on current endodontic knowledge and the classification of the problems are described. KEY LEARNING POINTS: * Thorough clinical and radiographic examination is required to diagnose and successfully treat minor to severe invaginations. * Modern clinical techniques may facilitate the management of invaginations once considered untreatable. PMID- 19133105 TI - 13th Biennial Congress of the European Society of Endodontology, Istanbul, Turkey, 6-8 september 2007. PMID- 19133107 TI - Bioinformatics: alive and kicking. AB - Bioinformatics has become too central to biology to be left to specialist bioinformaticians. Biologists are all bioinformaticians now. PMID- 19133108 TI - Recently published papers: Heavyweight problems in the intensive care unit? AB - The management of the obese patient in intensive care is fraught with difficulty, or so conventional wisdom would have us believe. Two recent studies add to the growing evidence that the obese patient is not significantly disadvantaged in terms of outcome following intensive care admission. Another study highlights the increasing burden of end-stage renal disease on intensive care unit resources with some interesting results. And there is more on citrate, the new anticoagulant! PMID- 19133109 TI - Next-generation biomass feedstocks for biofuel production. AB - The development of second-generation biofuels--those that do not rely on grain crops as inputs--will require a diverse set of feedstocks that can be grown sustainably and processed cost-effectively. Here we review the outlook and challenges for meeting hoped-for production targets for such biofuels in the United States. PMID- 19133110 TI - Chemotherapeutic treatment efficacy and sensitivity are increased by adjuvant alternating electric fields (TTFields). AB - BACKGROUND: The present study explores the efficacy and toxicity of combining a new, non-toxic, cancer treatment modality, termed Tumor Treating Fields (TTFields), with chemotherapeutic treatment in-vitro, in-vivo and in a pilot clinical trial. METHODS: Cell proliferation in culture was studied in human breast carcinoma (MDA-MB-231) and human glioma (U-118) cell lines, exposed to TTFields, paclitaxel, doxorubicin, cyclophosphamide and dacarbazine (DTIC) separately and in combinations. In addition, we studied the effects of combining chemotherapy with TTFields in an animal tumor model and in a pilot clinical trial in recurrent and newly diagnosed GBM patients. RESULTS: The efficacy of TTFields chemotherapy combination in-vitro was found to be additive with a tendency towards synergism for all drugs and cell lines tested (combination index 0.60) was found in HG chickens and in the wild junglefowl populations from Thailand. HG chickens exhibited the highest allelic richness (mean A = 2.9). No significant genetic subdivisions of the chicken population within the Ha Giang province were found. As compared to other breeds, HG chickens clustered with wild populations. Furthermore, the neighbornet tree and the Bayesian clustering analysis showed that chickens from 4 communes were closely related to the wild ones and showed an admixture pattern. CONCLUSION: In the absence of any population structuring within the province, the H'mong chicken, identified from its black phenotype, shared a common gene pool with other chickens from the Ha Giang population. The large number of alleles shared exclusively between Ha Giang chickens and junglefowl, as well as the results of a Bayesian clustering analysis, suggest that gene flow has been taking place from junglefowl to Ha Giang chickens. PMID- 19133137 TI - Terameprocol, a methylated derivative of nordihydroguaiaretic acid, inhibits production of prostaglandins and several key inflammatory cytokines and chemokines. AB - BACKGROUND: Extracts of the creosote bush, Larrea tridentata, have been used for centuries by natives of western American and Mexican deserts to treat a variety of infectious diseases and inflammatory disorders. The beneficial activity of this plant has been linked to the compound nordihydroguaiaretic acid (NDGA) and its various substituted derivatives. Recently, tetra-O-methyl NDGA or terameprocol (TMP) has been shown to inhibit the growth of certain tumor-derived cell lines and is now in clinical trials for the treatment of human cancer. In this report, we ask whether TMP also displays anti-inflammatory activity. TMP was tested for its ability to inhibit the LPS-induced production of inflammatory lipids and cytokines in vitro. We also examined the effects of TMP on production of TNF-alpha in C57BL6/J mice following a sublethal challenge with LPS. Finally, we examined the molecular mechanisms underlying the effects we observed. METHODS: RAW 264.7 cells and resident peritoneal macrophages from C57BL6/J mice, stimulated with 1 mug/ml LPS, were used in experiments designed to measure the effects of TMP on the production of prostaglandins, cytokines and chemokines. Prostaglandin production was determined by ELISA. Cytokine and chemokine production were determined by antibody array and ELISA.Western blots, q-RT-PCR, and enzyme assays were used to assess the effects of TMP on expression and activity of COX-2.q-RT-PCR was used to assess the effects of TMP on levels of cytokine and chemokine mRNA.C57BL6/J mice injected i.p. with LPS were used in experiments designed to measure the effects of TMP in vivo. Serum levels of TNF alpha were determined by ELISA. RESULTS: TMP strongly inhibited the production of prostaglandins from RAW 264.7 cells and normal peritoneal macrophages. This effect correlated with a TMP-dependent reduction in levels of COX-2 mRNA and protein, and inhibition of the enzymatic activity of COX-2.TMP inhibited, to varying degrees, the production of several cytokines, and chemokines from RAW 264.7 macrophages and normal peritoneal macrophages. Affected molecules included TNF-alpha and MCP-1. Levels of cytokine mRNA were affected similarly, suggesting that TMP is acting to prevent gene expression.TMP partially blocked the production of TNF-alpha and MCP-1 in vivo in the serum of C57BL6/J mice that were challenged i.p. with LPS. CONCLUSION: TMP inhibited the LPS-induced production of lipid mediators and several key inflammatory cytokines and chemokines, both in vitro and in vivo, raising the possibility that TMP might be useful as a treatment for a variety of inflammatory disorders. PMID- 19133140 TI - An experience of virtual leadership development for human resource managers. AB - PROBLEM: Strong leadership and management skills are crucial to finding solutions to the human resource crisis in health. Health professionals and human resource (HR) managers worldwide who are in charge of addressing HR challenges in health systems often lack formal education in leadership and management. APPROACH: Management Sciences for Health (MSH) developed the Virtual Leadership Development Program (VLDP) with support from the United States Agency for International Development (USAID). The VLDP is a Web-based leadership development programme that combines face-to-face and distance-learning methodologies to strengthen the capacity of teams to identify and address health challenges and produce results. RELEVANT CHANGES: The USAID-funded Leadership, Management and Sustainability (LMS) Program, implemented by MSH, and the USAID-funded Capacity Project, implemented by IntraHealth, adapted the VLDP for HR managers to help them identify and address HR challenges that ministries of health, other public-sector organizations and nongovernmental organizations are facing. LOCAL SETTINGS: Three examples illustrate the results of the VLDP for teams of HR managers: 1. the Uganda Protestant and Catholic Medical Bureaus 2. the Christian Health Association of Malawi 3. the Developing Human Resources for Health Project in Uganda. LESSONS LEARNT: The VLDP is an effective programme for developing the management and leadership capacity of HR managers in health. PMID- 19133139 TI - Absence of the complement regulatory molecule CD59a leads to exacerbated neuropathology after traumatic brain injury in mice. AB - BACKGROUND: Complement represents a crucial mediator of neuroinflammation and neurodegeneration after traumatic brain injury. The role of the terminal complement activation pathway, leading to generation of the membrane attack complex (MAC), has not been thoroughly investigated. CD59 is the major regulator of MAC formation and represents an essential protector from homologous cell injury after complement activation in the injured brain. METHODS: Mice deleted in the Cd59a gene (CD59a-/-) and wild-type littermates (n = 60) were subjected to focal closed head injury. Sham-operated (n = 60) and normal untreated mice (n = 14) served as negative controls. The posttraumatic neurological impairment was assessed for up to one week after trauma, using a standardized Neurological Severity Score (NSS). The extent of neuronal cell death was determined by serum levels of neuron-specific enolase (NSE) and by staining of brain tissue sections in TUNEL technique. The expression profiles of pro-apoptotic (Fas, FasL, Bax) and anti-apoptotic (Bcl-2) mediators were determined at the gene and protein level by real-time RT-PCR and Western blot, respectively. RESULTS: Clinically, the brain injured CD59a-/- mice showed a significantly impaired neurological outcome within 7 days, as determined by a higher NSS, compared to wild-type controls. The NSE serum levels, an indirect marker of neuronal cell death, were significantly elevated in CD59a-/- mice at 4 h and 24 h after trauma, compared to wild-type littermates. At the tissue level, increased neuronal cell death and brain tissue destruction was detected by TUNEL histochemistry in CD59a-/- mice within 24 hours to 7 days after head trauma. The analysis of brain homogenates for potential mediators and regulators of cell death other than the complement MAC (Fas, FasL, Bax, Bcl-2) revealed no difference in gene expression and protein levels between CD59a-/- and wild-type mice. CONCLUSION: These data emphasize an important role of CD59 in mediating protection from secondary neuronal cell death and further underscore the key role of the terminal complement pathway in the pathophysiology of traumatic brain injury. The exact mechanisms of complement MAC-induced secondary neuronal cell death after head injury require further investigation. PMID- 19133141 TI - Filtering for increased power for microarray data analysis. AB - BACKGROUND: Due to the large number of hypothesis tests performed during the process of routine analysis of microarray data, a multiple testing adjustment is certainly warranted. However, when the number of tests is very large and the proportion of differentially expressed genes is relatively low, the use of a multiple testing adjustment can result in very low power to detect those genes which are truly differentially expressed. Filtering allows for a reduction in the number of tests and a corresponding increase in power. Common filtering methods include filtering by variance, average signal or MAS detection call (for Affymetrix arrays). We study the effects of filtering in combination with the Benjamini-Hochberg method for false discovery rate control and q-value for false discovery rate estimation. RESULTS: Three case studies are used to compare three different filtering methods in combination with the two false discovery rate methods and three different preprocessing methods. For the case studies considered, filtering by detection call and variance (on the original scale) consistently led to an increase in the number of differentially expressed genes identified. On the other hand, filtering by variance on the log2 scale had a detrimental effect when paired with MAS5 or PLIER preprocessing methods, even when the testing was done on the log2 scale. A simulation study was done to further examine the effect of filtering by variance. We find that filtering by variance leads to higher power, often with a decrease in false discovery rate, when paired with either of the false discovery rate methods considered. This holds regardless of the proportion of genes which are differentially expressed or whether we assume dependence or independence among genes. CONCLUSION: The case studies show that both detection call and variance filtering are viable methods of filtering which can increase the number of differentially expressed genes identified. The simulation study demonstrates that when paired with a false discovery rate method, filtering by variance can increase power while still controlling the false discovery rate. Filtering out 50% of probe sets seems reasonable as long as the majority of genes are not expected to be differentially expressed. PMID- 19133142 TI - Removal of spermatozoa with externalized phosphatidylserine from sperm preparation in human assisted medical procreation: effects on viability, motility and mitochondrial membrane potential. AB - BACKGROUND: Externalization of phosphatidylserine (EPS) occurs in apoptotic-like spermatozoa and could be used to remove them from sperm preparations to enhance sperm quality for assisted medical procreation. We first characterized EPS in sperms from infertile patients in terms of frequency of EPS spermatozoa as well as localization of phosphatidylserine (PS) on spermatozoa. Subsequently, we determined the impact of depleting EPS spermatozoa on sperm quality. METHODS: EPS were visualized by fluorescently-labeled annexin V binding assay. Double staining with annexin V and Hoechst differentiates apoptotic from necrotic spermatozoa. We used magnetic-activated cell sorting using annexin V-conjugated microbeads (MACS ANMB) technique to remove EPS spermatozoa from sperm prepared by density gradient centrifugation (DGC). The impact of this technique on sperm quality was evaluated by measuring progressive motility, viability, and the integrity of the mitochondrial membrane potential (MMP) by Rhodamine 123. RESULTS: Mean percentages of EPS spermatozoa were 14% in DGC sperm. Four subpopulations of spermatozoa were identified: 70% alive, 3% early apoptotic, 16% necrotic and 11% late apoptotic or necrotic. PS were localized on head and/or midpiece or on the whole spermatozoa. MACS efficiently eliminates EPS spermatozoa. MACS combined with DGC allows a mean reduction of 70% in EPS and of 60% in MMP-disrupted spermatozoa with a mean increase of 50% in sperm survival at 24 h. CONCLUSION: Human ejaculates contain EPS spermatozoa which can mostly be eliminated by DGC plus MACS resulting in improved sperm long term viability, motility and MMP integrity. EPS may be used as an indicator of sperm quality and removal of EPS spermatozoa may enhance fertility potential in assisted medical procreation. PMID- 19133143 TI - Expression of human papilloma virus type 16 E5 protein in amelanotic melanoma cells regulates endo-cellular pH and restores tyrosinase activity. AB - BACKGROUND: Melanin synthesis, the elective trait of melanocytes, is regulated by tyrosinase activity. In tyrosinase-positive amelanotic melanomas this rate limiting enzyme is inactive because of acidic endo-melanosomal pH. The E5 oncogene of the Human Papillomavirus Type 16 is a small transmembrane protein with a weak transforming activity and a role during the early steps of viral infections. E5 has been shown to interact with 16 kDa subunit C of the trans membrane Vacuolar ATPase proton pump ultimately resulting in its functional suppressions. However, the cellular effects of such an interaction are still under debate. With this work we intended to explore whether the HPV16 E5 oncoprotein does indeed interact with the vacuolar ATPase proton pump once expressed in intact human cells and whether this interaction has functional consequences on cell metabolism and phenotype. METHODS: The expression of the HPV16-E5 oncoproteins was induced in two Tyrosinase-positive amelanotic melanomas (the cell lines FRM and M14) by a retroviral expression construct. Modulation of the intracellular pH was measured with Acridine orange and fluorescence microscopy. Expression of tyrosinase and its activity was followed by RT-PCR, Western Blot and enzyme assay. The anchorage-independence growth and the metabolic activity of E5 expressing cells were also monitored. RESULTS: We provide evidence that in the E5 expressing cells interaction between E5 and V ATPase determines an increase of endo-cellular pH. The cellular alkalinisation in turn leads to the post-translational activation of tyrosinase, melanin synthesis and phenotype modulation. These effects are associated with an increased activation of tyrosine analogue anti-blastic drugs. CONCLUSION: Once expressed within intact human cells the HPV16-E5 oncoprotein does actually interact with the vacuolar V-ATPase proton pump and this interaction induces a number of functional effects. In amelanotic melanomas these effects can modulate the cell phenotype and can induce a higher sensitivity to tyrosine related anti-blastic drugs. PMID- 19133144 TI - Modulation of human uterine smooth muscle cell collagen contractility by thrombin, Y-27632, TNF alpha and indomethacin. AB - BACKGROUND: Preterm labour occurs in approximately 10% of pregnancies and is a major cause of infant morbidity and mortality. However, the pathways involved in regulating contractility in normal and preterm labour are not fully elucidated. Our aim was to utilise a human myometrial contractility model to investigate the effect of a number of uterine specific contractility agents in this system. Therefore, we investigated the contractile response of human primary uterine smooth muscle cells or immortalised myometrial smooth muscle cells cultured within collagen lattices, to known mediators of uterine contractility, which included thrombin, the ROCK-1 inhibitor Y-27632, tumour necrosis factor alpha (TNF alpha) and the non-steroidal anti-inflammatory indomethacin. METHODS: Cell contractility was calculated over time, with the collagen gel contraction assay, utilising human primary uterine smooth muscle cells (hUtSMCs) and immortalised myometrial smooth muscle cells (hTERT-HM): a decrease in collagen gel area equated to an increase in contractility. RNA was isolated from collagen embedded cells and gene expression changes were analysed by real time fluorescence reverse transcription polymerase chain reaction. Scanning electron and fluorescence microscopy were employed to observe cell morphology and cell collagen gel interactions. Statistical analysis was performed using ANOVA followed by Tukey's post hoc tests. RESULTS: TNF alpha increased collagen contractility in comparison to the un-stimulated collagen embedded hUtSMC cells, which was inhibited by indomethacin, while indomethacin alone significantly inhibited contraction. Thrombin augmented the contractility of uterine smooth muscle cell and hTERT-HM collagen gels, this effect was inhibited by the thrombin specific inhibitor, hirudin. Y-27632 decreased both basal and thrombin-induced collagen contractility in the hTERT-HM embedded gels. mRNA expression of the thrombin receptor, F2R was up-regulated in hUtSMCs isolated from collagen gel lattices, following thrombin stimulated contractility. CONCLUSION: TNF alpha and thrombin increased uterine smooth muscle cell collagen contractility while indomethacin had the opposite effect. Thrombin-induced collagen contractility resulted in F2R activation which may in part be mediated by the ROCK-1 pathway. This study established the in vitro human myometrial model as a viable method to assess the effects of a range of uterotonic or uterorelaxant agents on contractility, and also permits investigation of the complex regulatory pathways involved in mediating myometrial contractility at labour. PMID- 19133147 TI - Locked lung by looped hernia: a case report. AB - BACKGROUND: Large pleural effusions are usually symptomatic. We report a patient with asymptomatic massive left sided pleural effusion with left lung collapse secondary to a traumatic diaphragmatic hernia. CASE PRESENTATION: A 44 year old male presented with recurrent pleural effusions over six weeks. His pleural effusion was first diagnosed incidentally on a chest X-ray after a fall. Extensive diagnostic studies were unyielding for the etiology of the effusion. A diagnostic and therapeutic video assisted thoracoscopy revealed a diaphragmatic hernia with inflamed, incarcerated omentum. After hernia repair there was no recurrence. CONCLUSION: This case underscores the obscure presentation of an incarcerated diaphragmatic hernia presenting as massive recurrent pleural effusions. PMID- 19133148 TI - Acute thrombosis in mitralic mechanical prosthesis: a case report. AB - We describe a case of a man, 42 years old, submitted to successful fibrinolitic strategy with rTPA in acute mitralic prosthesis valve thrombosis and ipomobility of one emidisk. There aren't a consensus agreement in therapeutic strategy but we may support the approach of some authors that employ fibrinolisis in patients without absolute or relative controindications and if thrombus dimension is less than 1 cm otherwise they make use of heparin therapy in non obstructive thrombosis with successive transesophageal echocardiography evaluation for the efficacy and the later indication for thrombolisis or surgery treatment. PMID- 19133149 TI - Cost analysis of an integrated disease surveillance and response system: case of Burkina Faso, Eritrea, and Mali. AB - BACKGROUND: Communicable diseases are the leading causes of illness, deaths, and disability in sub-Saharan Africa. To address these threats, countries within the World Health Organization (WHO) African region adopted a regional strategy called Integrated Disease Surveillance and Response (IDSR). This strategy calls for streamlining resources, tools, and approaches to better detect and respond to the region's priority communicable disease. The purpose of this study was to analyze the incremental costs of establishing and subsequently operating activities for detection and response to the priority diseases under the IDSR. METHODS: We collected cost data for IDSR activities at central, regional, district, and primary health care center levels from Burkina Faso, Eritrea, and Mali, countries where IDSR is being fully implemented. These cost data included personnel, transportation items, office consumable goods, media campaigns, laboratory and response materials and supplies, and annual depreciation of buildings, equipment, and vehicles. RESULTS: Over the period studied (2002-2005), the average cost to implement the IDSR program in Eritrea was $0.16 per capita, $0.04 in Burkina Faso and $0.02 in Mali. In each country, the mean annual cost of IDSR was dependent on the health structure level, ranging from $35,899 to $69,920 at the region level, $10,790 to $13,941 at the district level, and $1,181 to $1,240 at the primary health care center level. The proportions spent on each IDSR activity varied due to demand for special items (e.g., equipment, supplies, drugs and vaccines), service availability, distance, and the epidemiological profile of the country. CONCLUSION: This study demonstrates that the IDSR strategy can be considered a low cost public health system although the benefits have yet to be quantified. These data can also be used in future studies of the cost-effectiveness of IDSR. PMID- 19133150 TI - Thrombus or tumor? a case of fibroelastoma as indicated during the submission process. AB - We describe the case of a 50-year-old woman who was admitted to a pheriferal department for heart failure. The echocardiography revealed a small mass measuring about 1.3 x 1.0 cm adhering to the non-coronary cusp of the aortic valve, mild dilated cardiomiopathy and severe biventricular dysfunction. This mass had erroneously been considered a thrombotic lesion, so the patient was treated with thrombolysis and heparin e.v. Only after a transoesophageal echocardiography a tumour cardiac mass was suspected. The diagnosis of fibroelastoma was confirmed by MRI and then from the anatomic and histoligical definition after surgery. PMID- 19133146 TI - A tool for self-assessment of communication skills and professionalism in residents. AB - BACKGROUND: Effective communication skills and professionalism are critical for physicians in order to provide optimum care and achieve better health outcomes. The aims of this study were to evaluate residents' self-assessment of their communication skills and professionalism in dealing with patients, and to evaluate the psychometric properties of a self-assessment questionnaire. METHODS: A modified version of the American Board of Internal Medicine's (ABIM) Patient Assessment survey was completed by 130 residents in 23 surgical and non-surgical training programs affiliated with a single medical school. Descriptive, regression and factor analyses were performed. Internal consistency, inter-item gamma scores, and discriminative validity of the questionnaire were determined. RESULTS: Factor analysis suggested two groups of items: one group relating to developing interpersonal relationships with patients and one group relating to conveying medical information to patients. Cronbach's alpha (0.86) indicated internal consistency. Males rated themselves higher than females in items related to explaining things to patients. When compared to graduates of U.S. medical schools, graduates of medical schools outside the U.S. rated themselves higher in items related to listening to the patient, yet lower in using understandable language. Surgical residents rated themselves higher than non-surgical residents in explaining options to patients. CONCLUSION: This appears to be an internally consistent and reliable tool for residents' self-assessment of communication skills and professionalism. Some demographic differences in self-perceived communication skills were noted. PMID- 19133152 TI - The role of ENSO in understanding changes in Colombia's annual malaria burden by region, 1960-2006. AB - BACKGROUND: Malaria remains a serious problem in Colombia. The number of malaria cases is governed by multiple climatic and non-climatic factors. Malaria control policies, and climate controls such as rainfall and temperature variations associated with the El Nino/Southern Oscillation (ENSO), have been associated with malaria case numbers. Using historical climate data and annual malaria case number data from 1960 to 2006, statistical models are developed to isolate the effects of climate in each of Colombia's five contrasting geographical regions. METHODS: Because year to year climate variability associated with ENSO causes interannual variability in malaria case numbers, while changes in population and institutional control policy result in more gradual trends, the chosen predictors in the models are annual indices of the ENSO state (sea surface temperature [SST] in the tropical Pacific Ocean) and time reference indices keyed to two major malaria trends during the study period. Two models were used: a Poisson and a Negative Binomial regression model. Two ENSO indices, two time reference indices, and one dummy variable are chosen as candidate predictors. The analysis was conducted using the five geographical regions to match the similar aggregation used by the National Institute of Health for its official reports. RESULTS: The Negative Binomial regression model is found better suited to the malaria cases in Colombia. Both the trend variables and the ENSO measures are significant predictors of malaria case numbers in Colombia as a whole, and in two of the five regions. A one degree Celsius change in SST (indicating a weak to moderate ENSO event) is seen to translate to an approximate 20% increase in malaria cases, holding other variables constant. CONCLUSION: Regional differentiation in the role of ENSO in understanding changes in Colombia's annual malaria burden during 1960-2006 was found, constituting a new approach to use ENSO as a significant predictor of the malaria cases in Colombia. These results naturally point to additional needed work: (1) refining the regional and seasonal dependence of climate on the ENSO state, and of malaria on the climate variables; (2) incorporating ENSO-related climate variability into dynamic malaria models. PMID- 19133151 TI - Hypothesis: primary antiangiogenic method proposed to treat early stage breast cancer. AB - BACKGROUND: Women with Down syndrome very rarely develop breast cancer even though they now live to an age when it normally occurs. This may be related to the fact that Down syndrome persons have an additional copy of chromosome 21 where the gene that codes for the antiangiogenic protein Endostatin is located. Can this information lead to a primary antiangiogenic therapy for early stage breast cancer that indefinitely prolongs remission? A key question that arises is when is the initial angiogenic switch thrown in micrometastases? We have conjectured that avascular micrometastases are dormant and relatively stable if undisturbed but that for some patients angiogenesis is precipitated by surgery. We also proposed that angiogenesis of micrometastases very rarely occurs before surgical removal of the primary tumor. If that is so, it seems possible that we could suggest a primary antiangiogenic therapy but the problem then arises that starting a therapy before surgery would interfere with wound healing. RESULTS: The therapy must be initiated at least one day prior to surgical removal of the primary tumor and kept at a Down syndrome level perhaps indefinitely. That means the drug must have virtually no toxicity and not interfere meaningfully with wound healing. This specifically excludes drugs that significantly inhibit the VEGF pathway since that is important for wound healing and because these agents have some toxicity. Endostatin is apparently non-toxic and does not significantly interfere with wound healing since Down syndrome patients have no abnormal wound healing problems. CONCLUSION: We propose a therapy for early stage breast cancer consisting of Endostatin at or above Down syndrome levels starting at least one day before surgery and continuing at that level. This should prevent micrometastatic angiogenesis resulting from surgery or at any time later. Adjuvant chemotherapy or hormone therapy should not be necessary. This can be continued indefinitely since there is no acquired resistance that develops, as happens in most cancer therapies. PMID- 19133153 TI - Simple allele-discriminating PCR for cost-effective and rapid genotyping and mapping. AB - BACKGROUND: Single nucleotide polymorphisms (SNPs) are widely observed between individuals, ecotypes, and species, serving as an invaluable molecular marker for genetic, genomic, ecological and evolutionary studies. Although, a large number of SNP-discriminating methods are currently available, few are suited for low throughput and low-cost applications. Here, we describe a genotyping method named Simple Allele-discriminating PCR (SAP), which is ideally suited for the small scale genotyping and gene mapping routinely performed in small to medium research or teaching laboratories. RESULTS: We demonstrate the feasibility and application of SAP to discriminate wild type alleles from their respective mutant alleles in Arabidopsis thaliana. Although the design principle was previously described, it is unclear if the method is technically robust, reliable, and applicable. Three primers were designed for each individual SNP or allele with two allele discriminating forward primers (one for wild type and one for the mutant allele) and a common reverse primer. The two allele-discriminating forward primers are designed so that each incorporates one additional mismatch at the adjacent (penultimate) site from the SNP, resulting in two mismatches between the primer and its non-target template and one mismatch between the primer and its target template. The presence or absence of the wild type or the mutant allele correlates with the presence or absence of respective PCR product. The presence of both wild type-specific and mutant-specific PCR products would indicate heterozygosity. SAP is shown here to discriminate three mutant alleles (lug-3, lug-16, and luh-1) from their respective wild type alleles. In addition, the SAP principle is shown to work in conjunction with fluorophore-labeled primers, demonstrating the feasibility of applying SAP to high throughput SNP analyses. CONCLUSION: SAP offers an excellent alternative to existing SNP-discrimination methods such as Cleaved Amplified Polymorphic Sequence (CAPS) or derived CAPS (dCAPS). It can also be adapted for high throughput SNP analyses by incorporating fluorophore-labeled primers. SAP is reliable, cost-effective, fast, and simple, and can be applied to all organisms not limited to Arabidopsis thaliana. PMID- 19133154 TI - Accessory nipple reconstruction following a central quadrantectomy: a case report. AB - INTRODUCTION: nipple dichotomy (or intra-areolar polythelia) is a rare congenital malformation in which one or more supernumerary nipples are located within the same areola.A case of a woman undergoing a central quadrantectomy with a contralateral supernumerary nipple used for reconstruction is reported. No other report in the Literature, according to our search, has focused on reconstructive use of an accessory nipple after breast conserving surgery. CASE PRESENTATION: the patient is a 73 year-old Caucasian woman, who two years earlier underwent a lower-outer left Quadrantectomy plus axillary sampling and radiation therapy for a 2,2 cm lobular carcinoma with no lymph node involvement.A routine follow-up assessment showed an important fibrotic change on the operated breast, just across the infra-mammary fold; at a breast Magnetic Resonance Imaging, a 1,5 cm area in retroareolar position, suspicious for local recurrence, was evident.An open biopsy was therefore performed, under local anaesthesia, including the nipple-areolar complex to realize a central Quadrantectomy with a Grisotti procedure; a congenital dichotomic nipple in the contralateral breast was then used to repair the defect through a "nipple-sharing" technique. The final histological examination reported a fibrotic mastopathy without atypias. CONCLUSION: in this case, the "nipple-sharing" technique has allowed in the same time the correction of a rare congenital defect and provided the surgeon with a supernumerary nipple to be used in the immediate reconstruction after breast conserving surgery. PMID- 19133155 TI - Association between maternal use of traditional healer services and child vaccination coverage in Pont-Sonde, Haiti. AB - BACKGROUND: Child vaccination is one of the public health interventions that are responsible for the relatively low child morbidity and mortality in developed nations compared to the developing world. We carried out this study to examine the association between mothers' use of traditional healer services and vaccination among Haitian children. Our hypothesis was that children whose mothers used the services of traditional healers were less likely to be vaccinated compared to children whose mothers did not use the services of traditional healers. METHODS: A two-stage stratified sampling method was used to select 720 mothers from the population of Pont-Sonde, Haiti. Of these mothers, 691 (96%) completed the survey by responding to a standardized questionnaire on vaccination giving unadjusted odds ratios (OR) and adjusted odds ratios (AOR) and 95% confidence intervals (CI) and use of traditional healers. Bivariate and multivariate logistic regression analyses were performed to estimate the effect of explanatory variables on vaccination (the main outcome). RESULTS: Mother's use of traditional healer services was negatively associated with vaccination after controlling for maternal age, education, religion, and distance from the nearest health care facility. For those children whose mothers often or always used the services of traditional healers, we found a 53% decrease in the odds of vaccination (AOR = 0.47; 95% CI [0.27, 0.83]) compared against children whose mothers never used the services of the traditional healers. There were negative associations between practice of Vodou and vaccination (AOR = 0.56; 95% CI [0.35, 0.92]), and distance from the nearest health care service facility and vaccination (AOR = 0.53; 95% CI [0.29, 0.97] and AOR = 0.34; 95% CI [0.20, 0.59] at 46-60 and more than 60 minutes walk time, respectively). CONCLUSION: We found that mother's use of traditional healer services was negatively associated with vaccination of Haitian children. Findings from this study underscore the potential to enlist the support of traditional healers in promoting child health by educating, mentoring them (the traditional healers) in supporting vaccination efforts. PMID- 19133156 TI - Preoperative Y-90 microsphere selective internal radiation treatment for tumor downsizing and future liver remnant recruitment: a novel approach to improving the safety of major hepatic resections. AB - BACKGROUND: Extended liver resections are being performed more liberally than ever. The extent of resection of liver metastases, however, is restricted by the volume of the future liver remnant (FLR). An intervention that would both accomplish tumor control and induce compensatory hypertrophy, with good patient tolerability, could improve clinical outcomes. CASE PRESENTATION: A 53-year-old woman with a history of cervical cancer presented with a large liver mass. Subsequent biopsy indicated poorly differentiated carcinoma with necrosis suggestive of squamous cell origin. A decision was made to proceed with pre operative chemotherapy and Y-90 microsphere SIRT with the intent to obtain systemic control over the disease, downsize the hepatic lesion, and improve the FLR. A surgical exploration was performed six months after the first SIRT (three months after the second). There was no extrahepatic disease. The tumor was found to be significantly decreased in size with central and peripheral scarring. The left lobe was satisfactorily hypertrophied. A formal right hepatic lobectomy was performed with macroscopic negative margins. CONCLUSION: Selective internal radiation treatment (SIRT) with yttrium-90 (Y-90) microspheres has emerged as an effective liver-directed therapy with a favorable therapeutic ratio. We present this case report to suggest that the portal vein radiation dose can be substantially increased with the intent of inducing portal/periportal fibrosis. Such a therapeutic manipulation in lobar Y-90 microsphere treatment could accomplish the end points of PVE with avoidance of the concern regarding tumor progression. PMID- 19133157 TI - A prospective, non-randomized phase II trial of Trastuzumab and Capecitabine in patients with HER2 expressing metastasized pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is the fourth most common cause of cancer related death in Western countries. Advantages in surgical techniques, radiation and chemotherapy had almost no impact on the long term survival of affected patients. Therefore, the need for better treatment strategies is urgent. HER2, a receptor tyrosine kinase of the EGFR family, involved in signal transduction pathways leading to cell growth and differentiation is overexpressed in a number of cancers, including breast and pancreatic cancer. While in breast cancer HER2 has already been successfully used as a treatment target, there are only limited data evaluating the effects of inhibiting HER2 tyrosine kinases in patients with pancreatic cancer. METHODS: Here we report the design of a prospective, non randomized multi-centered Phase II clinical study evaluating the effects of the Fluoropyrimidine-carbamate Capecitabine (Xeloda) and the monoclonal anti-HER2 antibody Trastuzumab (Herceptin) in patients with non-resectable, HER2 overexpressing pancreatic cancer. Patients eligible for the study will receive Trastuzumab infusions on day 1, 8 and 15 concomitant to the oral intake of Capecitabine from day 1 to day 14 of each three week cycle. Cycles will be repeated until tumor progression. A total of 37 patients will be enrolled with an interim analysis after 23 patients. DISCUSSION: Primary end point of the study is to determine the progression free survival after 12 weeks of bimodal treatment with the chemotherapeutic agent Capecitabine and the anti-HER2 antibody Trastuzumab. Secondary end points include patient's survival, toxicity analysis, quality of life, the correlation of HER2 overexpression and clinical response to Trastuzumab treatment and, finally, the correlation of CA19-9 plasma levels and progression free intervals. PMID- 19133159 TI - How 40 kilograms of fluid retention can be overlooked: two case reports. AB - INTRODUCTION: With a rising incidence of severe obesity in developed nations, heart failure, a well-recognised co-morbidity, is becoming more common. CASE PRESENTATION: We describe two recent patients encountered, a 64 year old and a 42 year old, who are both severely obese and presented with fluid retention of approximately 40 kilograms. Assessment revealed that the explanation of the gross clinical features were relatively subtle cardiac abnormalities. These cases illustrate how fluid retention in severe obesity can differ from that seen in 'traditional' heart failure in terms of clinical assessment and management. CONCLUSION: Severe obesity can result in insidious fluid retention, which can be easily overlooked until large volumes of fluid have accumulated. Cardiac abnormalities are usually found in these patients, but may be relatively subtle, leading to current debate in the definition and classification of heart failure. These scenarios are increasingly being encountered in clinical practice. Recognition, assessment and treatment of the 'clinical syndrome of heart failure' in severe obesity is often difficult. PMID- 19133158 TI - Multiple splice defects in ABCA1 cause low HDL-C in a family with hypoalphalipoproteinemia and premature coronary disease. AB - BACKGROUND: Mutations at splice junctions causing exon skipping are uncommon compared to exonic mutations, and two intronic mutations causing an aberrant phenotype have rarely been reported. Despite the high number of functional ABCA1 mutations reported to date, splice variants have been reported infrequently. We screened DNA from a 41 year-old male with low HDL-C (12 mg/dL [0.31 mmol/L]) and a family history of premature coronary heart disease (CHD) using polymerase chain reaction single-strand conformation polymorphism (SSCP) analysis. METHODS: Family members with low levels of HDL-C (n = 6) were screened by SSCP for mutations in ABCA1. Samples with altered SSCP patterns were sequenced directly using either an ABI 3700 or ABI3730Xl DNA Analyzer. To screen for splicing defects, cDNA was isolated from the proband's RNA and was sequenced as above. A series of minigenes were constructed to determine the contribution of normal and defective alleles. RESULTS: Two novel splice variants in ABCA1 were identified. The first mutation was a single base pair change (T->C) in IVS 7, 6 bps downstream from the exon7/intron7 junction. Amplification of cDNA and allelic subcloning identified skipping of Exon 7 that results in the elimination of 59 amino acids from the first extracellular loop of the ABCA1 protein. The second mutation was a single base pair change (G->C) at IVS 31 -1, at the intron/exon junction of exon 32. This mutation causes skipping of exon 32, resulting in 8 novel amino acids followed by a stop codon and a predicted protein size of 1496 AA, compared to normal (2261 AA). Bioinformatic studies predicted an impact on splicing as confirmed by in vitro assays of constitutive splicing. CONCLUSION: In addition to carnitine-acylcarnitine translocase (CACT) deficiency and Hermansky-Pudlak syndrome type 3, this represents only the third reported case in which 2 different splice mutations has resulted in an aberrant clinical phenotype. PMID- 19133160 TI - Studying alternative splicing regulatory networks through partial correlation analysis. AB - BACKGROUND: Alternative pre-mRNA splicing is an important gene regulation mechanism for expanding proteomic diversity in higher eukaryotes. Each splicing regulator can potentially influence a large group of alternative exons. Meanwhile, each alternative exon is controlled by multiple splicing regulators. The rapid accumulation of high-throughput data provides us with a unique opportunity to study the complicated alternative splicing regulatory network. RESULTS: We propose the use of partial correlation analysis to identify association links between exons and their upstream regulators or their downstream target genes (exon-gene links) and links between co-spliced exons (exon-exon links). The partial correlation analysis avoids taking the ratio of two noisy random variables, exon expression level and gene expression level, so that it achieves a higher statistical power. We named this analysis procedure pCastNet (partial Correlation analysis of splicing transcriptome Network). Through studies of known alternative exons, conservation patterns, relative positions, functional annotations, and RT-PCR experiments, we concluded that pCastNet can effectively identify exon-gene or exon-exon links. We further found that gene pairs with exon gene or exon-exon links tend to have similar functions or are present in the same pathways. More interestingly, gene pairs with exon-gene or exon-exon links tend to share cis-elements in promoter regions and microRNA binding elements in 3' untranslated regions, which suggests the coupling of co-alternative-splicing, co transcription-factor-binding, and co-microRNA-binding. CONCLUSIONS: Alternative splicing regulatory networks reconstructed by pCastNet can help us better understand the coordinate and combinatorial nature of alternative splicing regulation. The proposed tool can be readily applied to other high-throughput data such as transcriptome sequencing data. PMID- 19133162 TI - Pure sensory stroke from compression of putaminal haemorrhage: a case report. AB - INTRODUCTION: The literature rarely describes putaminal haemorrhage producing pure spinothalamic sensory deficit. Here reports a case of putaminal haemorrhage in which selective impairment of the spinothalamic sensory modality was due to the compression of the hematoma. CASE PRESENTATION: A 57 year old hypertensive man presented with a pure sensory stroke(PSS), and CT scan showed a putaminal haemorrhage. The clinical course was characterized by rapid resolution of the deficits. CONCLUSION: This case illustrates this rarely of PSS from compression of putaminal haemorrhage of good functional and vital prognosis, and stresses the value of CT scanning for diagnosis and prognosis. PMID- 19133163 TI - Explaining cross-country variation in cigarette consumption. AB - This short paper uses cross-country data on per capita cigarette consumption and selected socio-economic variables to explain inter-country differentials in consumption. It is found that the proportion of the aged in the total population and higher literacy among women have relatively greater and positive impact on cigarette consumption. Even after controlling for the effect of the two variables, a country's industrialized status has a positive impact on consumption. It would thus seem that aging and economic, and social developments are pro-cigarette consumption. PMID- 19133161 TI - Human Rad51 mediated DNA unwinding is facilitated by conditions that favour Rad51 dsDNA aggregation. AB - BACKGROUND: Human Rad51 (RAD51), analogous to its bacterial homolog, RecA, binds and unwinds double stranded DNA (dsDNA) in the presence of certain nucleotide cofactors. ATP hydrolysis is not required for this process, because even ATP non hydrolysable analogs like AMP-PNP and ATPgammaS, support DNA unwinding. Even ADP, the product of ATP hydrolysis, feebly supports DNA unwinding. RESULTS: We find that human Rad52 (RAD52) stimulates RAD51 mediated DNA unwinding in the presence of all Adenine nucleotide cofactors, (except in AMP and no nucleotide conditions that intrinsically fail to support unwinding reaction) while enhancing aggregation of RAD51-dsDNA complexes in parallel. Interestingly, salt at low concentration can substitute the role of RAD52, in facilitating aggregation of RAD51-dsDNA complexes, that concomitantly also leads to better unwinding. CONCLUSION: RAD52 itself being a highly aggregated protein perhaps acts as scaffold to bring together RAD51 and DNA molecules into large co-aggregates of RAD52-RAD51-DNA complexes to promote RAD51 mediated DNA unwinding reaction, when appropriate nucleotide cofactors are available, presumably through macromolecular crowding effects. Our work highlights the functional link between aggregation of protein-DNA complexes and DNA unwinding in RAD51 system. PMID- 19133164 TI - Role of Nurr1 and Ret in inducing rat embryonic neural precursors to dopaminergic neurons. AB - BACKGROUND: Nurr1, a member of the nuclear receptor superfamily of transcription factors, is highly expressed in midbrain dopaminergic (DA) neurons. Ret is a member of the receptor tyrosine kinase (RTK) superfamily and a critical signal transducing subunit of receptors for glial cell line-derived neurotrophic factor (GDNF). Both Nurr1 and Ret play important roles in the development of DA neurons. PURPOSE: To investigate possible correlation between Nurr1 and Ret on inducing expression of tyrosine hydroxylase (TH) in neural precursor cells. METHODS: Neural precursors isolated from rat embryonic mesencephalon (E13.5d) were transfected with vectors containing Nurr1 or Ret and treated with 9-cis-retinoic acid (RA) and/or GDNF for 3 days. RT-PCR and immunocytochemistry was used to test the expression of Nurr1 and Ret and the TH positive cells. RESULTS: The number of TH positive cells was increased from 1.53 +/- 0.12 to 3.83 +/- 0.56% after the cells were transfected with Nurr1. Increased endogenous Nurr1 by RA lead to 1.8 times (2.86 +/- 0.32% versus 1.53 +/- 0.12% in the controls) increase in TH positive cells. A double inducing effect by both endogenous and exogenous Nurr1 on the expression of TH was observed by 3.3 times increase in the positive cells (from 5.03 +/- 0.76 to 1.53 +/- 0.12% in control). Ret expression was induced by overexpression of Nurr1. Overexpressed Ret had no inducing effect on the expression of Nurr1 and the number of TH positive cells. The gene of dopamine transporter (DAT) was clearly induced in the cells transfected with Ret. CONCLUSION: Nurr1, required for the expression of Ret, had inducing effect on TH positive cells, and Ret may promote maturation of DA neuron by up-regulating DAT expression through its ligand. As a cooperator, Ret seems to work together with Nurr1 in the development of DA neurons. PMID- 19133165 TI - Peri-operative medical management of cerebral vasospasm. AB - INTRODUCTION: The physiological mechanism of cerebral vasospasm after aneurysmal subarachnoid hemorrhage remains elusive and its treatment can be challenging. Traditionally, 'triple-H' therapy and the calcium channel blocker, nimodipine, are used to treat cerebral vasospasm. However, as the etiology of vasospasm is unraveled, investigative pharmaceutical agents that stop the development and attenuate the severity of cerebral vasospasm, are being investigated in clinical trials. METHODS: In this manuscript, we review the clinical presentation and characteristics of cerebral vasospasm after aneurysmal subarachnoid hemorrhage and the utility of hyperdynamic therapy and pharmacotherapies. RESULTS: Triple-H therapy improves cerebral perfusion and improves neurological outcome during clinically evident cerebral vasospasm. Nimodipine is the accepted standard medication used to reduce the incidence of cerebral vasospasm, but more importantly, has a neuroprotective effect during hypoxia. Other medications such as magnesium sulfate, HMG-CoA reductase inhibitors and enoxaparin, are also being trialed with some promising results. CONCLUSION: Endovascular administration of intra-arterial anti-spasmodic agents and balloon angioplasty are becoming more commonly utilized at institutions where endovascular therapy is available. However, triple-H therapy and nimodipine remain the accepted first-line of treatment for cerebral vasospasm after aneurysmal subarachnoid hemorrhage. PMID- 19133166 TI - Peri-operative measures for treatment and prevention of cerebral vasospasm following subarachnoid hemorrhage. AB - OBJECTIVE: Aneurysmal subarachnoid hemorrhage (aSAH) is associated with high early mortality rates. Cerebral vasospasm remains the major source of morbidity after aSAH. Angiographic evidence of vasospasm is apparent in 70% of patients, while clinical manifestation of vasospasm is present in one third of patients. Early or existing vasospasm at the time of presentation poses an additional challenge in the management of the patient, and forms the basis for this review. METHODS: Treatment modalities for management of ruptured aneurysms in the setting of vasospasm, including timing of aneurysm surgery and peri-operative management, are reviewed. Intraoperative measures aimed at treatment of existing vasospasm and at the prevention of vasopasm are discussed. RESULTS: Operative/endovascular means to secure the ruptured aneurysm should be performed as soon as possible to facilitate treatment of the vasospasm. Surgery performed in the presence of angiographic/symptomatic vasospasm can be associated with good outcome. Operative measures to decrease the incidence of vasospasm include clot removal, intracisternal injection of thrombolytics, fenestration of the lamina terminalis and local application of vasodilatory agents. Post-operative measures include early intra-arterial injection of vasodilators (verapamil or nicardipine), percutaneous angioplasty, triple-H therapy and CSF drainage. DISCUSSION: The utilization of a multimodality approach to treat patients with aneurysmal subarachnoid hemorrhage presenting with existing vasospasm can result in good outcome. PMID- 19133167 TI - Familial and genetic researches on three Chinese families with von Hippel-Lindau disease. AB - OBJECTIVE: Hemangioblastoma of the central nervous system (CNS) occur as sporadic tumors or as a part of von Hippel-Lindau (VHL) disease, an autosomal dominant hereditary tumor syndrome caused by germline mutation of the VHL tumor suppressor gene. This study shows the clinical characteristics of three large Chinese families with VHL disease and evaluates the consequence of the genetic test for the diagnosis of VHL disease and clinical screening of the family members. METHODS: DNA is extracted from peripheral blood in 43 members from three large families with VHL disease and amplified by PCR to three exons of the VHL gene. The PCR products were directly sequenced and the mutations compared with the Human Gene Mutation Database. RESULTS: The ages of the patients who are given the initial diagnosis ranged from 16 to 47 years (mean: 31 years), and the mean time was 17.3 months (2-30 months) from the emergence of the symptom to patients' first visit. Furthermore, the gender distribution was 20% female (4) and 80% male (16). Twenty VHL disease patients in the three families have the most common manifestation of CNS hemangioblastoma. The cytosine replaced the 716th guanine on four patients and three carriers of virulence gene from the first family, which made the 168th serine replaced by threonine. And no mutation was found on the 22 members of the second family. Meanwhile, it was also found that the guanine replaced the 559th cytosine on one patient and two carriers from the third family, which made the 116th leucine replaced by valine. CONCLUSION: The DNA analysis of VHL germline mutations is clearly superior to clinical information to diagnose VHL disease. The CNS hemangioblastoma is the early manifestation in VHL disease. It is recommended that every patient with CNS hemangioblastoma should be screened for VHL gene mutation. The test for the VHL gene plays a key role in the discovery of asymptomatic patients and the carriers of virulence gene. PMID- 19133170 TI - [Psychosocial risk factors for illicit drug use in a sample of Mexican high school students]. AB - OBJECTIVES: To identify psychosocial risk factors for substance abuse among Mexican students and to offer elements for the design of prevention programs. METHODS: A cross-sectional, nonexperimental study of a sample of 516 high school students in six of Mexico's most important cities. From April-June 2005, a customized version of the Drug Use Screening Inventory (revised) (DUSI-R) was administered. The analysis comprised eight factors: alcohol and drug abuse, affective disorders, poor self-control, poor school adjustment, low social competence, dysfunctional family relationships, social isolation, and being part of a detrimental social network (whose members take drugs and have antisocial attitudes). Factors predictive for illicit drug use were found by logistical regression, and a structural equation model was designed to determine the relationships among the factors. RESULTS: The factors that predicted substance abuse were poor self-control with a tendency to act impulsively and aggressively; associating with troublemakers; and being frequently exposed to family conflicts, violence, and drug and/or alcohol use in the home. The structural equation model indicated that substance abuse is one of a group of disorders directly determined by associating with detrimental peers, and a higher rate of socioaffective disorders, and indirectly, by dysfunctional family relationships. CONCLUSIONS: Some of the suggestions made by theoretical models to explain substance abuse were confirmed. These empirically-supported elements can contribute to the design of prevention programs, especially those that are selective and recommended. PMID- 19133168 TI - Carotid bruit for detection of hemodynamically significant carotid stenosis: the Northern Manhattan Study. AB - OBJECTIVE: The prevalence of carotid bruits and the utility of auscultation for predicting carotid stenosis are not well known. We aimed to establish the prevalence of carotid bruits and the diagnostic accuracy of auscultation for detection of hemodynamically significant carotid stenosis, using carotid duplex as the gold standard. METHODS: The Northern Manhattan Study (NOMAS) is a prospective multiethnic community-based cohort designed to examine the incidence of stroke and other vascular events and the association between various vascular risk factors and subclinical atherosclerosis. Of the stroke-free cohort (n=3298), 686 were examined for carotid bruits and underwent carotid duplex. Main outcome measures included prevalence of carotid bruits and sensitivity, specificity, positive predictive value, negative predictive value and accuracy of auscultation for prediction of ipsilateral carotid stenosis. RESULTS: Among 686 subjects with a mean age of 68.2 +/- 9.4 years, the prevalence of >/=60% carotid stenosis as detected by ultrasound was 2.2% and the prevalence of carotid bruits was 4.1%. For detection of carotid stenosis, sensitivity of auscultation was 56%, specificity was 98%, positive predictive value was 25%, negative predictive value was 99% and overall accuracy was 97.5%. DISCUSSION: In this ethnically diverse cohort, the prevalence of carotid bruits and hemodynamically significant carotid stenosis was low. Sensitivity and positive predictive value were also low, and the 44% false-negative rate suggests that auscultation is not sufficient to exclude carotid stenosis. While the presence of a bruit may still warrant further evaluation with carotid duplex, ultrasonography may be considered in high-risk asymptomatic patients, irrespective of findings on auscultation. PMID- 19133172 TI - [Agricultural toxic use and temporal distribution of male birth rate in the state of Parana, Brazil]. AB - OBJECTIVE: To determine the trend in male birth rates from 1994-2004 in the state of Parana, Brazil, and whether a correlation exists between this trend and the state's agricultural toxic sales in 1985. METHOD: This ecological study employed data from the Brazilian Institute for Geography and Statistics (agricultural toxic sales) and the Unified Health System's database "DATASUS" (sex ratio at birth). Three hundred and eight (308) municipalities were analyzed, divided into quartiles by pesticide usage. Each of the four resulting quartiles included 77 municipalities, with the fourth quartile representing the highest consumption. The male birth rate was obtained for each year for each municipality. Then the median rate was calculated for the entire period (1994-2004) for each municipality. A bivariate analysis was carried out. Pearson coefficients were calculated for each quartile. An exploratory and comparative analysis of quartiles was performed, and the mean per capita consumption was calculated for each quartile. Ten municipalities with intense agricultural activity were then selected and analyzed individually in terms of their agricultural profile and male birth rate trend during the study period. RESULTS: A discrete decline was observed in the rate of live male births for the entire state. The magnitude of this decline was more evident when the group of 10 municipalities was analyzed: in this group, the rates observed at the end of the study period were much lower than those usually described in the literature (male birth rate < 50%). Pearson's correlation was negative for all quartiles, although without statistical significance. CONCLUSIONS: The statistically significant trend towards a decline in male births in some of the Parana municipalities suggests that this phenomenon may be a result of a high level of environmental exposure to pesticides. Future studies should confirm and measure the impact of environmental pesticide contamination on reproductive health, expressed in this study as sex ratio at birth. PMID- 19133171 TI - Pharmacoeconomics of antidepressants in moderate-to-severe depressive disorder in Colombia. AB - OBJECTIVE: To compare three antidepressant drugs from different classes used in treating moderate-to-severe major depressive disorder (MDD) in Colombian adults. METHODS: Based on expert input, a decision-tree model was adapted for Colombia to analyze data over 6 months from the government-payer perspective. The cost effectiveness of amitriptyline, fluoxetine, and venlafaxine was determined. The clinical outcome was remission of depression (a score 2 h; 29.7 % applied pesticides against the wind, while 26.4 % applied these with the wind. After applying the pesticides, 37.4 % of them bathed after 3 h, 34.5 % changed clothes, and 18.8 % returned to cultivation some hours later; 23 % of workers presented some symptom, the most frequent being headache, itchiness, numbness, and perception of burning on skin; this latter proportion was greater in the < or = 46 years old group CONCLUSIONS: symptom prevalence is in agreement with inappropriate pesticide management; nonetheless, we found no statistically significant association. Our results are lower than other studies, possibly due to lesser exposure, determined by toxicity, pesticide amount utilized, and use and management of chemicals. PMID- 19133186 TI - [The influence of different factors on energy intake and Basal metabolic rate in children with cancer]. AB - OBJECTIVE: to identify the Influence of different factors on energy intake and basal metabolic rate in children with cancer at diagnosis. METHODS: the basal metabolic rate and energy intake were measured during hospitalization and before treatment. The basal metabolic rate and energy from foods were measured by indirect calorimetry. Data were compared by Student t test and a multiple linear regression. RESULTS: energy intake ranged from 636 to 3063 kcal/d, mean 1956 +/- 530 kcal/d. Measured basal metabolic rate was within 10 % of predict for Schofield equation in ten of fourteen patients. Four patients were classified as hypometabolic. Energy intake was related to kind of tumor (solid tumor), sex (males) and loss weight, while basal metabolic rate was related to the age and sex, but only the age reached statistical significance (p < 0.01). CONCLUSIONS: the most influent variables on basal metabolic rate and energy intake were the variables related with age, sex and nutritional status at diagnosis. Physician should be consider the energy requirements of children in order to establish an appropriate nutritional and oncology therapy, and consequently, modify the risk of nutritional deterioration in order to improve the prognostic of patients. PMID- 19133187 TI - [Pneumoconiosis in affiliated workers have irreversible sequelae]. AB - OBJECTIVE: to analyze the frequency and importance of pneumoconiosis in affiliated workers to Instituto Mexicano del Seguro Social. MATERIAL AND METHODS: it was carried out a transverse, observational and comparative study; the behavior of the qualified pneumoconiosis was revised 1994-2004. The analyzed variables were sex, age, occupation, permanent disability and economic activity. It was carried out the analysis of absolute frequencies and rates. RESULTS: it was registered 14,827 cases; the rate for 10,000 workers oscillated from 0.48 to 2.39. The 99 % were presented in males, with 14,262 cases with permanent disability (rate: 96.89 per 100 cases of pneumoconiosis); employments with more frequency were miners, quarry workers and bricklayers. The average of permanent disability for pneumoconiosis in the period was of 11.09 to 15 %. States with a higher number of cases were Coahuila, Zacatecas and Hidalgo. Economic activities with a greater incidence were the extraction and benefit of mineral coal, graphite and metallic and non-metallic minerals. CONCLUSION: cases of pneumoconiosis have been decreasing, but even the determination of diagnosis is delayed, and due to this most workers have irreversible sequelae. PMID- 19133188 TI - [Physician's humanistic qualities expected by society]. AB - Forming physicians educative institutions must pay attention to society expectations about health professionals. Physicians and patients as users of health services were interviewed in order to know what the physician's profile is to be qualified as satisfactory. Results were analyzed with a qualitative method. Thirty seven health professionals were interviewed. Group included: physicians, nurses, social medical workers, medical office assistants and medicine students. We interviewed three patient groups (hypertensive and diabetic patients) and 30 additional patients with age range from 18 to 50, most of them female. Characteristics that were more frequently mentioned by patients: skills in communication in order to give them better information about its disease, kindly and humane care (compassion and emphatic). These aspects are related more with attitude than to knowledge. Therefore, we can conclude that forming physician institutions should not worry only their graduate's cognitive skills, but also to define their humane qualities. This would make possible to establish suitable pedagogic strategies in order to achieve and assess humane formation to fit with society expectations. PMID- 19133189 TI - [Urethral duplication with giant calculus. Case report]. AB - BACKGROUND: the urethral duplication is an uncommon congenital anomaly. We presented a case of male patient with congenital urethral duplication with a giant calculus in the accessory urethra. CASE: male patient 23 years old with sensation of tumor and pain in the lower pelvis and base of the penis with decrease of the urinary spurt and final dripping. There was edema in the penescrotal area; a hard tumor can be touched, painful to movement. The urethrography and cistoscopy showed urethral duplication type IIB according to the classification of Effman without communication with bladder, it had rooted before the fossa navicular in a blind-ending which was hidden at the bulbar urethral where the giant calculus was located. A surgical procedure was done to remove it. It measured 3 x 2 cm. The patient had a satisfactory recuperation. CONCLUSION: the presence of a giant calculus in the urethral is a strange situation and when it does happen, one must suspect malformation in such structure. There was no surgical correction done to the urethral duplication. The etiological factor for the formation of the giant calculus seems to be the urinary stasis, it is usually present in the urethral duplication. The surgical treatment must be individualized for each patient according to type of anatomical duplication. PMID- 19133190 TI - [Disc alterations of lumbar spine on magnetic resonance images in asymptomatic workers]. AB - OBJECTIVE: to determine abnormal findings of the lumbar spine on magnetic resonance images in asymptomatic subjects. METHODS: prospective, transverse and descriptive study, in workers of the Instituto Mexicano del Seguro Social without low back pain; they were invited to be observed with magnetic resonance images of lumbar spine. A total of 105 cases was interpreted by a radiologist, who did not know the patients' clinical conditions. 107 lumbar spine alterations studies were mixed in order to not influence in the results, and they were not included in the statistic analysis. RESULTS: 55 % of the cases had discal alterations, 38 % presented bulging disk and 17 % presented protrusion. Other alterations were Schmorl's nodule, osteocondrosis, espondilolistesis, and annular tears. CONCLUSIONS: bulging disk and discal protrusion frequency have high prevalence in magnetic resonance images in healthy individuals, so its presence in symptomatic patients is not necessarily cause of low back pain. PMID- 19133191 TI - [Ambulatory treatment of prolonged pregnancy with prostaglandin E2 gel]. AB - BACKGROUND: the rutinary labor induction with prostaglandin E2 (PGE2) in pregnancy at 41 weeks has showed a fetal benefit without an increase in maternal morbidity or cesarean delivery. OBJECTIVE: to show that the ambulatory management of prolonged pregnancy with PGE2 gel decreases the cesarean delivery and prenatal morbidity rates. METHODS: quasiexperimetal study of patients with an accurate dated pregnancy of 41 weeks and beyond were analyzed. The women were divided in two groups of 196 each one. In the treated group, the endocervical application of PGE2 was followed by cardiotocographic control. If there was no reason to interrupt the pregnancy they were evaluated twice a week. Perinatal outcomes, mode of delivery and indications for cesarean section were assessed in both groups. RESULTS: there was a decrease in rate of cesarean delivery in treated group, 43 % versus 54 % in control group (p < 0.05). Apgar score at 1 and 5 minute showed no difference, but there were two intrauterine deaths in control group. The indications for cesarean surgery were the same in both groups and there was a case of tachysystole in each one. CONCLUSION: we concluded that decrease in the rate of cesarean deliveries without increments of fetal and maternal morbidity in this study, point to a secure management choice with PGE2 in ambulatory patients. PMID- 19133192 TI - [Four years experience in the follow up of nursing children with urolithiasis]. AB - OBJECTIVE: to describe clinical data, metabolic profile, treatment and evolution of a nursing children cohort with urolithiasis. METHODS: using a descriptive, prospective and longitudinal design, were registered all patients with urolithiasis, of Pediatric Division at the Hospital General "Agustin O'Horan," in Yucatan, Mexico. We included all patients who were 36 months old and younger. We did not include children with malformations or associated disease which causes urolithiasis. Clinical and metabolic profiles, evaluation of the applied treatment as well as an evolution by four years period were done. RESULTS: 63 patients were recruited with ages ranging from 2 to 36 months; 39 (61.9 %) were male. The most common clinical signs were crying upon urination (49.2 %), urinary retention (31.7 %), hematuria (30.2 %), and pollakiuria (28.6 %). Calculus were most frequently located in the kidney (58.7 %). The most common complications were acute urinary tract infection (61.9 %), malnutrition (47.6 %) and acute renal failure (12.7 %). Documented metabolic alterations included hyperuricosuria (57.1 %), hypomagnesiuria (11.1 %) and hypercalciuria (9.5 %). CONCLUSIONS: urolithiasis should be suspected in children with crying upon urination and urinary symptoms; it should be considered among the different causes of renal failure; hyperuricosuria was the most frequent metabolic alteration. PMID- 19133193 TI - [Neural defect prevalence in 248,352 consecutive newborns]. AB - BACKGROUND: neural tube defects are a scourge for the population in nondeveloped countries. OBJECTIVE: to evaluate the prevalence of neural tube defects at a Gynecology and Obstetric Hospital at Instituto Mexicano del Seguro Social. METHODS: it was compared two 5 years periods: one before (1995-1999) and other after (2000-2004) a State Health Department program (Secretaria de Salud, Nuevo Leon) with 5 mg folic acid supplementation once a week to women in reproductive age. RESULTS: we registered 248,352 consecutive deliveries in a 10 year period, and we observed 319 cases of neural tube defects, with a global rate of 12.84 per 10,000 deliveries. From 1995 to 1999, there were 132,360 deliveries, with 212 neural tube defects (16.01 rate per 10,000 deliveries); the rate of anencephaly was 6.49, spina bifida 8.23, and encephalocele 1.28. After the folic acid program was established, during the following period 2000 to 2004, there were 115,992 deliveries, 108 with neural tube defects (9.31 rate per 10,000 newborns), with 41.8 % reduction (p <0.001). Also decreased were anencephaly rates: 28.3 % (nonsignificant) and spina bifida 63.4 % (p < 0.001). CONCLUSIONS: the neural tube defects rate has been declining in the last five years compared with the 5 years previous by 41 %, significantly more for spina bifida than anencephaly. PMID- 19133194 TI - [Gas-forming liver abscess in diabetic patient]. AB - Pyogenic liver abscess is still considered a medical challenge due to the seriousness of the clinical course, late diagnosis, and usually high mortality. Gas-forming liver abscess is not uncommon, the incidence among all pyogenic liver abscess is around 7 to 24 %, mostly in individuals in their sixth and seventh decades ,and with diabetes mellitus disease. The epidemiology of gas-forming liver abscess has been changing with a shift from young to older diabetic adults. A clinical case of a 71 year old diabetic female with gas-forming liver abscess is presented; we describe the clinical course and treatment is analyzed. PMID- 19133195 TI - [Maternal mortality decrease at IMSS, 2000-2005. As a result of specific actions or by chance?]. AB - OBJECTIVE: to assessment the epidemiological characteristics and evolution of maternal hospital deaths at Instituto Mexicano del Seguro Social (IMSS) on the years 2000-2005 and to analyze a possible relation with a specific program to reduce it, starting in 2001. METHODS: we collected and studied data about 253 and 144 hospital maternal deaths between 2000 and 2005, respectively. We compared rates, causes, type of obstetrical death, age, parity, history of prenatal care and preventability at admission in hospitals were the women died. Data about live births in were obtained from the official Information Medical System of IMSS. The analysis was made with descriptive statistical measures and values of chi(2). RESULTS: the maternal mortality rate fell by 30.8 %, as result of a decline from 39 per 100,000 live births in 2000, to 27 in 2005; the proportion of direct obstetric deaths showed a reduction from 77.1 to 66.7 %; all the specific-age mortality rates felt too, but with some variability. Preeclampsia-eclampsia, hemorrhages and abortion were responsible for more of 50 % of total deaths in both years compared. We did not observe significant changes in other variables. CONCLUSIONS: the reduction in general rate of hospital maternal mortality, in percentage of direct obstetric causes of death and specific-age rates were chronologically coincidental with the development of a program to increase opportunity and quality of obstetric care at the whole institution. The results let us think there is a possible cause-effect relation. It is imperative to conduct a more long-term observation to confirm this epidemiological phenomenon. PMID- 19133196 TI - [A brief historical account of IMSS hospital in Orizaba, Veracruz]. AB - In 1947, Orizaba City was chosen, due its political, economic and labor importance, for the Instituto Mexicano del Seguro Social (IMSS) to establish its health service in Veracruz State. At the beginning, hospital care was rented out at the Hospital Civil "Ignacio de la Llave." In 1959 a hospital was specifically constructed and had 280 beds. In 1973 a earthquake partially destroyed the hospital building and the medical services were almost discontinued. In 1975 it was rebuilt, and was renamed "Ignacio Garcia Tellez," who had been a founder member of IMSS. At the beginning the main causes of hospital admissions were infectious diseases and gynecologic and obstetric problems; at present the challenges are the chronic diseases and accidents. PMID- 19133197 TI - [Health actions to become integrated into IMSS-oportunidades]. AB - The Program IMSS-Oportunidades has a 28 year history in the public health arena in Mexico, serving the indigenous and most marginalized population living in 17 204 localities in the country. With 3548 health units and 69 hospitals the program delivers health and community interventions providing better health services to this historically underserved population. The program rests in the work developed by 269,000 volunteers including traditional healers. In general terms there were 20 million medical visits to the health units, 215,000 hospital discharges and nearly 85,000 surgeries. Preventive programs are important and evaluated through the immunization coverages, early detection of several diseases and the descending numbers of communicable diseases. The main causes of death show a mixture of infectious and chronic diseases where cardiovascular disease and diabetes are highlighted. This panorama gives a brief summary of the efforts displayed by the program and an institution that distinguishes for its level of organization and efficiency. PMID- 19133198 TI - [Risk cardiovascular factors, clinical epidemiology and disease]. PMID- 19133199 TI - [Prevalence of buccal lesions on type 2 diabetes mellitus]. AB - OBJECTIVE: to know the prevalence of buccal lesions on type 2 diabetes mellitus patients and its association with blood glycemia levels. METHODS: descriptive cross-sectional study. Casual glycemia was practiced to 162 patients, who regularly attend public health services, and results from fasting blood glucose test were analyzed. A buccal examination and World Health Organization's community periodontal index and treatment needs (CPITN) was practiced. Descriptive statistics and bivaried analysis were used. RESULTS: the woman:man relation was 3:1, age range from 17 to 80 years. Out of 99 patients with laboratory glycemia, 76 % was poorly controlled; 99 % showed at least one buccal lesion: 93.2 %, periodontal disease; 73.4 %, xerostomia and 28.4 %, erythematous candidiasis. Having bad controlled glucose, a group with xerostomia and erythematous candidiasis and another showed xerostomia plus erythematous candidiasis plus periodontal disease (OR = 3.5 and a value of p = 0.01). CONCLUSIONS: presence of oral diseases is exhibited on patients with a poor glucose control. Medical attention results inefficient and insufficient and patients do not get orientation to understand their illness. PMID- 19133200 TI - [Lipid profile in household contacts of patients with pulmonary tuberculosis]. AB - BACKGROUND: we have recently published the hypothesis that hypocholesterolemia might be a risk factor for the development of pulmonary tuberculosis. In this sense, albeit patients with pulmonary tuberculosis often have total cholesterol levels lower than the general population, it is not known if the latter have had a recent exposure to Mycobacterium tuberculosis. OBJECTIVE: to evaluate serum lipids in subjects exposed to mycobacteria. METHODS: we studied 25 pulmonary tuberculosis patients and 44 household contacts, and compared their serum lipid profile. RESULTS: We found that total cholesterol, LDL and triglycerides concentrations increased with age in contacts but not in pulmonary tuberculosis patients, with statistically significant differences in regression lines (age versus lipid level). Multiple linear regression analysis confirmed that being a household contact was associated with higher levels of total cholesterol, LDL, HDL and triglycerides. CONCLUSIONS: We conclude that lipid profile differed between pulmonary tuberculosis patients and their household contacts, thus supporting that low cholesterol levels might be a risk factor for developing pulmonary tuberculosis. PMID- 19133201 TI - [Associated family factors with nutrition and oral health in adolescents]. AB - OBJECTIVE: to evaluate the association of family factors with malnutrition and oral condition in adolescence. METHODS: a cross-sectional study of 456 adolescents was carried out to examine associations between family factors and risk for overweight or overweight (ROoO), risk for short stature or short stature (RSSoSS), and a DMFT (decayed, filled and missing-teeth) index > or = 8. RESULTS: the risk factors for ROoO were maternal educational level of high school or above (OR = 1.9, IC 95 % = 1.16-3.14) and having less than two siblings (OR = 1.6, IC 95 % = 1.13-2.34). Age > 15 years is associated with a DMFT index > or = 8 (OR = 1.9, IC 95 % = 1.14-2.34). CONCLUSIONS: a higher maternal educational level and having one or no siblings were ROoO risks. Increased age was a risk factor for caries. PMID- 19133202 TI - [Tomography-electroencephalography agreement in pediatric patients with epilepsy]. AB - OBJECTIVE: to evaluate the tomography-electroencephalography agreement in pediatric patients attended with epilepsy diagnosis at service neuropediatric in the Medical Unit of High Specialization "Adolfo Ruiz Cortines," during the period from January to October of year 2006. METHODS: retrospective, cross-sectional, descriptive, correlational, observational. One hundred eighty files of pediatric patients were reviewed with clinical diagnosis of epilepsy with electroencephalography of date next to their study of tomography; the agreement was realized with the test of kappa, comparing the results of each diagnosis method, in each patient. RESULTS: of 69 patients, 10 patients were catalogued as normal by both methods and 23 showed cerebral alterations in the same site of epilepsy focus; the observed agreement was 47.8 % with an index of kappa of 0.095 CONCLUSION: the agreement between the electroencephalography and the computed tomography is very low, in the findings of cerebral structural injuries related to the location of the epilepsy focus in pediatric patients with epilepsy diagnosis. PMID- 19133203 TI - [Validation of three screening tests used for early detection of cervical cancer]. AB - OBJECTIVE: to evaluate the validity (sensitivity, specificity, and accuracy) of three screening methods used in the early detection of the cervical carcinoma versus the histopathology diagnosis. METHODS: a selected sample of 107 women attended in the Opportune Detection of Cervicouterine Cancer Program in the Hospital de Zona 46, Instituto Mexicano del Seguro Social in Durango, during the 2003 was included. The application of Papa-nicolaou, acetic acid test, and molecular detection of human papillomavirus, and histopatholgy diagnosis were performed in all the patients at the time of the gynecological exam. The detection and tipification of the human papillomavirus was performed by polymerase chain reaction (PCR) and analysis of polymorphisms of length of restriction fragments (RFLP). Histopathology diagnosis was considered the gold standard. The evaluation of the validity was carried out by the Bayesian method for diagnosis test. RESULTS: the positive cases for acetic acid test, Papanicolaou, and PCR were 47, 22, and 19. The accuracy values were 0.70, 0.80 and 0.99, respectively. CONCLUSION: since the molecular method showed a greater validity in the early detection of the cervical carcinoma we considered of vital importance its implementation in suitable programs of Opportune Detection of Cervicouterino Cancer Program in Mexico. However, in order to validate this conclusion, cross-sectional studies in different region of country must be carried out. PMID- 19133204 TI - [Cardiovascular risk factors associated to abdominal obesity in apparently healthy subjects]. AB - OBJECTIVE: to determine the relationship between the abdominal obesity and cardiovascular risk factors in apparently healthy subjects from Mexico City. METHODS: a total of 186 apparently healthy men and nonpregnant women from Mexico City, were enrolled in a cross-sectional study. A detailed medical history and physical examination were performed. Abdominal obesity was defined by waist circumference > or = 80 cm for women and > or = 90 cm for men. RESULTS: a total of 125 women (67.2 %) and 61 men (32.8 %) were enrolled. Of them, 151 (81.2 %) had insulin resistance and 130 (69.9 %) abdominal obesity. Among obese subjects 96 (46.2 %) showed metabolic syndrome. There were a high prevalence of hypertriglyceridemia (31 %) and low serum levels of HDL-cholesterol (58 %). CONCLUSIONS: the used cut point for abdominal obesity, despite identifying a high proportion of subjects with cardiovascular risk, did not recognize a high proportion of subjects with disorders in their lipid profile. PMID- 19133205 TI - [Medical knowledge validity: an approximation]. AB - The fulfillment of their professional responsibilities as necessity, prevent to physicians sometimes to reflect upon medical scientific knowledge validity. In this paper some reflections about it are made considering the importance of integral vision of medicine for the physicians. This knowledge result of scientific development and special application methods. The reflection on medical scientific knowledge validity is different from its historical or ethical valuation. The author point out four dimensions strongly connected on such validity: construct which result of theorical and empirical validity, and consensus validity. The four dimensions are modified for diverse factors. PMID- 19133206 TI - [Cardiovascular risk of cyclooxygenase selective inhibitors]. AB - It has been estimated that 30 million people worldwide take an nonsteroidal antiinflammatory drug (NSAID) daily. The main clinical objectives of these drugs are both to reduce joint pain and to improve joint function. However, gastrointestinal adverse events lead to the development of cyclooxygenase selective inhibitors (COXIB) with a better gastrointestinal safety profile. Since 1999, COXIB shown capacity to develop cardiovascular adverse events. Subsequent discoveries confirm overall risk of cardiovascular events. The increased cardiovascular risk occurred both in patients who were taking aspirin and in those who were not. Similar results with different COXIB appears to be a class effect of the COX-2 inhibitors, so patient risk factors must be identified and used in treatment decision making. Patients with gastrointestinal risk factors and no cardiovascular risk may benefit from use of a gastroprotective agent plus a nonselective nonsteroidal antiinflammatory drugs as a COXIB. We review assays whose objective was to study cardiovascular security of nonsteroidal antiinflammatory drugs and COXIB for known advantages and limitations of these drugs. PMID- 19133207 TI - [Letter to the editor]. PMID- 19133208 TI - [Effectiveness of a strategy in type 2 diabetic patient education in relation to metabolic control, nutrition and blood pressure]. AB - BACKGROUND: diet is a main strategy treatment in the diabetic patient. It allows effective use of energy sources in the tissues, and to support apropiate glucose, protein and lipid serum levels. OBJECTIVE: to determine the effectiveness of a participative educational program on metabolic control of patients with newly diagnosed diabetes mellitus type 2 (less than 12 months). METHODS: cuasiexperimental design with newly diagnosed diabetic patients of both sexes and previously informed consent. Biochemical and anthropometric markers were analyzed at basal measure and then monthly for 6 months: lipids profile, glycemia, glycosilated hemoglobin, body mass index, and tricipital fold and hip circumference and blood pressure. Educational sessions were with a participative strategy. The diet was calculated according with their physical activitiy and patient's requirements. A subsequent adjustments were done to due according biochemical measures. RESULTS: of 16 patients, 81.2 % were female. The average age was 52 +/- 1 3 years. There were statistical significance values in glucose (28.7 %), glycosilated hemoglobin (35.4 %), cholesterol (12.6 %), high density lipoproteins (11.3 %), systolic (6.1 %) and diastolic (16.8 %) blood pressure, body mass index (6.7 %) and weight (6.9 %). CONCLUSIONS: it was demonstrated that a participative educational program is effective in the control of newly diagnosed type 2 diabetic patient when diet is individualized according to requirements. PMID- 19133209 TI - [Acromegaly. Report of 84 cases]. AB - BACKGROUND: acromegaly is a rare disease, it is characterized by elevation of the levels of growth hormone and Insulin-like growth factor 1 (IGF-1). The definitive treatment of this disease is surgical. OBJECTIVE: to know the importance of this clinical disease and to exhort to the medical community for opportune delivery to specialist, to prove diagnosis and to treat with opportunity in order to avoid complications. METHODS: the clinical files of the patients received at Endocrinology Service from 1995 to 2005 were reviewed. The data of serum growth hormone levels after a 75 g oral glucose tolerance test, tumor size, criteria for cure, co-morbidities, affectation of hormonal axes and visual field defects of operated patients and not operated patients, were collected. RESULTS: 84 patients were studied, 35 men and 49 women, with an average of 42 years of aged agreement to the Classification of Hardy Vezina, was greater number of cases in degree 2 (macro adenoma with local expansion of the chair). 35 patients with microadenomas: and 25 with macroadenomas were operated. The most frequent co morbidities were the alteration in the metabolism of carbohydrates and hypertension, and a frequent hormonal affectation was hypothyroidism; the rate of biochemical cure was 7 %. Discordant results with levels of growth hormone high and normal levels of IGF-1 were in 10 %. CONCLUSIONS: the present study confirms that the acromegaly is diagnosed late and it had been associated with co morbidities and bigger tumor size, that they have influence on the surgical treatment results, since the rate of success were low (7 %). PMID- 19133210 TI - [Hearing loss in attended patients in otorhinolaryngology service at IMSS in Guadalajara]. AB - OBJECTIVE: to know the etiology, topography, degree of hearing loss and coexisting morbidity in outpatient otorhinolaryngology service at Instituto Mexicano del Seguro Social, Guadalajara, Jalisco. METHODS: descriptive and transversal study of 393 patients older than 12 years, were studied during 12 months. They were sent to the otorhinolaryngology service from five hospitals in Guadalajara. Three population groups were made (adolescents, adults and elderly); medical history, otoscopy and conventional tone audiometry was carried out. RESULTS: the average age of patients was 51.2 (SD 4.2 years), 57.8 % were women and 58.5 % adults. Of the 786 examined ears, 91.9 % (95% CI = 89.8-93.7) had hearing loss; chronic suppurative otitis media with or without sensorineural factor, was the most common of hearing loss, affecting 25.4 % (95% CI = 22.3 28.7) of the 723 hearing loss ears; after mixed cochleopathies (21.8 %, 95% CI = 18.8-25). Sensorineural hearing loss occurred in 85.6 % (95% CI = 82.8-88) and 31 % (95% CI= 27.4-34.3) had moderate hearing loss. The most common coexisting morbidity were high blood pressure and diabetes mellitus. CONCLUSIONS: our data suggest that hearing loss in population study is a treatable pathology, this information will aid in the promotion of preventive measures for opportune detection of otitis and comorbidity and to implement detection campaigns in population of risk, schools and labor centers. PMID- 19133211 TI - [Lidocaine with epinephrine effect on arterial tension of children]. AB - OBJECTIVE: to determine the effect of the epinephrine-lidocaine anaesthetic solutions on blood pressure in healthy children subjected to short dentistry procedures. METHODS: we performed a prospective, observational and analytical study in 39 children with 7 and 8 years of age, who were subjected to dentistry treatment of short duration in a dentistry service. The blood pressure was measured in three times: previous to the infiltration of lidocaine to 2 % with epinephrine; 10 minutes later; at the end of procedure. RESULTS: the systolic blood pressure, like diastolic and the average increased after 10 minutes of the infiltration with the anesthetic-vasoconstrictor solution, but this difference was not significant (p = 0.39, 0.14 and 0.40). We observed an increase of the systolic (6.0 mm Hg, 95 % CI = 4.6-7.5, F = 14.0, p = 0.0001), diastolic (9.9 mm Hg, 95 % CI = 7.3-12.5; F = 26.0, p = 0.0001) and average arterial tension (7.3 mm Hg, 95 % IC = 5.8-8.8; F = 23, p = 0.0001) to the 10 minutes after the treatment. CONCLUSIONS: the use of the epinephrine-lidocaine anaesthetic solutions does not have significant effect on the blood pressure of children subjected to short dentistry procedures. PMID- 19133212 TI - [Respiratory tract fluid microbiology in an intensive care unit]. AB - OBJECTIVE: to know patterns of antimicrobial resistance of bacterial isolates from tracheal aspirates in an Intensive Care Unit and to evaluate the cases of ventilator-associated pneumonia. METHODS: antibiotic sensitivity test was done. A comparison was made between patients with nosocomial pneumonia reported by infection surveillance team against those reported by the attending physician with the infectious disease consultant. RESULTS: Pseudomonas aeruginosa was the bacteria most frequently reported with 134 isolates (26 %), 71 were multiple-drug resistant; followed by Staphylococcus with 122 isolates (24 %), of which 88 were S. aureus with 62 of them (70 %) methicillin-resistant. Enterobacteriaceae, E. coli, K. pneumoniae, E. cloacae, S. marcescens, as well as Acinetobacter sp. and S. maltophilia were occasionally isolated. Candida represented 17 % of the isolates. Three peaks of isolates of S. aureus and P. aeruginosa were identified during the two years of surveillance. There were differences in cases of ventilator associated pneumonia reported by the hospital based epidemiology team and the attending clinicians in collaboration with an Infectious disease consultant. CONCLUSIONS: prevalence of multiple-drug-resistant Pseudomonas aeruginosa (53 %) and methicillin resistant Staphylococcus (70 %) isolated from the airway is high in our Intensive Care Unit. Enterobacterias, Acinetobacter sp. and Stenotrophomonas maltophilia colonization are low in our Intensive Care Unit. PMID- 19133213 TI - [The quality control in the coagulation laboratory]. AB - This is a broad review of sources of error in the coagulation tests, all of which must be considered, evaluated and supervised to improve their quality. The analytical result of a laboratory examination is a scientific fact and has no medical meaning as such. It must be interpreted to become a medical finding. Recent improvement in equipment, reagents, and the fact that hemophilic and thrombotic events are considered the main causes of death are the principal reasons to prepare this article. The internal quality control program considers this fact because its beginning starts when the physician makes the request to the lab; moreover, the clinical interpretation of the results of the laboratory represents the end of the cycle. Outcome of clinical test in hemostasis is critically dependent upon the quality of the sample, the method, the instruments, the reagents and the personnel involved in the performance of the test. Use of high quality blood collection procedures and an awareness of preanalytic errors are presented to lead to quality outcomes. Today, performance of internal and external quality control programs is obligatory. PMID- 19133214 TI - [Introduction. A brief history of AIDS]. AB - Human immunodeficiency virus (HIV) infection shows that knowledge of the molecular mechanisms involved in the pathogenesis can lead to the rapid development of new drugs to treat infected patients. Since the description of acquired immunodeficiency syndrome (AIDS) in 1981 and the identification of its causal agent in 1983, twenty-three antiretrovirals, belonging to 5 families, have been marketed. The viral entry point is one of the preferred therapeutic treatment targets, but the development of antagonists against the different HIV receptors has been a long process and fraught with difficulties. Maraviroc is the first CCR5 antagonist drug approved for clinical use and represents a milestone in the development of new treatments against HIV infection. Maraviroc is a novel drug and different from the rest of the antiretrovirals due to the special characteristics of its mechanism of action and is also the first antiretroviral directed towards a cell target. The different aspects of treatment with Maraviroc are analysed in this article; mechanism of action, toxicity, efficacy, resistance mechanisms, and its role in the context of antiretroviral treatment of the HIV infected patient. PMID- 19133215 TI - [Viral entry as therapeutic target. Current situation of entry inhibitors]. AB - Viral entry is an early stage and specific of the infection in which different viral and cellular targets are accessible to therapeutic treatment. CXCR4 and CCR5 act in this process as coreceptor molecules of HIV for its entry into the host cell. The predominant role played by the CCR5 coreceptor in the transmission and spreading of HIV makes this molecule the target of choice for blocking this mechanism. In the last few years, different specific inhibitors of HIV coreceptors have been generated of which only one, Maraviroc, has been approved for clinical use. The synthetic inhibitors developed act as allosteric antagonists that induce a non-permissive state or configuration of the coreceptor for binding viral envelope-glycoproteins. The CCR5 antagonists act on a wide spectrum of viruses with affinity or tropism for this receptor (virus R5), are absorbed orally and have powerful antiviral activity. However, the optimistic perspective offered by these new molecules has to be moderated due to the possible and expected appearance of viral resistances, on the one hand, and the propagation of viral species with affinity or tropism for the CXCR4 receptor (virus X4). This situation is a reality verified in the first clinical trials with these drugs and they acutely and urgently show the need to have effective and non-toxic CXCR4 inhibitors available to block this alternative viral replication and escape route. PMID- 19133216 TI - [Pharmacokinetics, interactions and mechanism of action of maraviroc]. AB - Maraviroc (MVC, Celsentri) is an allosteric and reversible inhibitor of the CCR5 chemokine coreceptor. MVC is the first marketed CCR5 antagonist and the only oral entry inhibitor approved so far for the treatment of HIV infection. It has been approved for adults with previous antiretroviral exposure. MVC exclusively inhibits the replication of R5- tropic HIV-1 variants after binding to the transmembrane CCR5 receptor cavity. MVC is rapidly absorbed following oral administration, and plasma T(max) is achieved within 0.5- 4 hours after a 300 mg dose. Renal clearance is approximately 10-12 L/h. MVC is a substrate of the cytochrome P450 isoenzyme 3A4; therefore dose adjustments are required when co administrated with other drugs that induce or inhibit CYP3A4. In addition, MVC dose adjustments are advised in patients with renal failure (CLcr <80 ml/min) only if they receive CYP3A4 inhibitors. PMID- 19133217 TI - [Maraviroc efficacy in clinical studies on the development of the molecule]. AB - Maraviroc is the first co-receptor antagonist to be approved for use in HIV infected patient. In a phase II trial, in which the drug was administered as a single therapy, the most suitable maintenance dose was 300 mg, once or twice per day. The MOTIVATE double blind, placebo-compared studies, were carried out on patients infected by HIV-1 with R5 tropism and resistant to drugs from three families of retrovirals. Maraviroc two times per day achieved < 50 copies/mL in 45.5% of the patients compared to 16.7% in the placebo group (p < 0.001). The CD4+ lymphocyte count had a mean of 63 cells/mm(3) higher with Maraviroc. The drug was shown to be superior in all patient groups regardless of the baseline viral load, the baseline CD4+ lymphocyte count or the number of accompanying active drugs. In the study on patients infected by HIV with X4/dual/mixed tropism, Maraviroc, was not virologically effective, but did produce a CD4 increase higher than the placebo. Maraviroc was compared with Efavirenz in the study on patients with no previous treatment and with R5-tropic virus. At 48 weeks, the percentage of patients with a viral load of <50 copies/mL was 69.3% in the group that received Efavirenz and 65.3% in the Maraviroc group. In conclusion, Maraviroc has demonstrated its increased efficacy in patients with CCR5-tropic virus and a long history of antiretroviral use and failure, and in patients with no previous treatment. PMID- 19133218 TI - [Secondary effects of treatment with maraviroc and other CCR5 antagonists. Potential impact of the CCR5 blocker]. AB - Maraviroc is the first inhibitor of CCR5 co-receptors to be marketed as an antiretroviral. The pre-clinical studies and phase III trials have shown that it has a very favourable safety profile. No characteristic adverse effect of maraviroc has been identified. Unlike with aplaviroc, where its clinical development was stopped due to serious hepatoxicity, no increase in liver toxicity has been demonstrated in patients treated with maraviroc even if they are co-infected by hepatotropic virus. Nor was there any evidence of an increase in the incidence of neoplasms or serious infections in patients treated with maraviroc. In a study on naive patients, maraviroc produced nonsignificant changes in total cholesterol, LDL, HDL and triglycerides. Although CCR5 co receptors play a role in the immune response of the body, it has not been shown whether individuals homozygote for its deletion (delta-32 mutation) have an increased risk of serious infections, with the possible exception of encephalitis due to the West Nile virus. However, long-term follow up is required on patients treated with to be able to rule out any increased susceptibility to infections or neoplasms. PMID- 19133219 TI - [Mechanisms of resistance and failure of treatment with maraviroc]. AB - Maraviroc (MVC) is a new antagonist of the CCR5 coreceptor and is the first antiviral compound available that has a cell factor essential for HIV entry as a target. The information available from clinical studies with MVC suggests that the main cause of therapeutic failure is, more than the tropism change, the rapid selection of pre-existing strains with an affinity for CXCR4, not detected by the reference test. A recently developed tropism test with an improved sensitivity will help to detect the minority, but clinically significant, presence of strains that use CXCR4. Evidence of resistance to MVC has been shown in vivo in some patients. The mechanism of this resistance appears to be related to changes in gp120 and mainly in the V3 region which enables the virus to recognise the CCR5 co-receptor bound to the MVC molecule. From a practical point of view, standardised tests are currently unavailable to assess susceptibility to MVC, although in dose-response phenotype tests a maximum percentage inhibition (MPI) < 95% would be indicative of resistance to the compound. Similarly, although some mutations associated with resistance in V3, and other zones of gp120, have been described, this preliminary information suggests different resistance patterns and at the moment, we do not know the canonical mutations to be able to establish genotyping algorithms. PMID- 19133221 TI - [Methods for determining viral tropism: genotype and phenotype tests]. AB - The recent approval of the first CCR5 antagonist, Maraviroc (MVC, Celsentri), with specific antiviral activity against R5-tropic virus variants has generated the need for studies to determine the viral tropism in all those patient candidates for starting treatment with this new drug. Although genotyping methods appear to be the most useful tool due to its speed and simplicity, in the case of viral tropism, phenotyping techniques are currently considered the most reliable. In the last few years, different phenotyping assays have been developed to determine the use of the co-receptor. However, the Trofile phenotype assay is currently the one most used for the determination of tropism, since it is the only one that has been clinically validated. Given that the presence of X4- tropic variants in the viral population has been associated with virological failure to MVC, the main challenge of both genotyping and phenotyping tools is to optimise their sensitivity for detecting X4-tropic variants present in a minority of the viral population. At the same time, the correlation between genotyping/phenotyping methods is being evaluated to determine whether genotyping tools can be useful to make therapeutic decisions. PMID- 19133220 TI - [At what time and with which combinations should maraviroc be indicated in the new antiretroviral treatment scenario?]. AB - Maraviroc is a selective and slowly reversible antagonist of the CCR5 co-receptor which has shown to have powerful antiviral activity, in vitro, against a wide range of HIV clinical isolates, including strains multi-resistant to 4 classes of pre-existing antiretroviral drugs. Maraviroc is active against HIV populations that only use the CCR5 coreceptor to enter the cell and has not demonstrated significant activity in the treatment of viral populations that use the CXCR4 co receptor. The mechanism of action of maraviroc, non-competitive with other antiretroviral drugs, and the absence of crossed resistance with the rest of their families, has led to Maraviroc being a drug available for use in rescue antiretroviral treatment. However, the excellent tolerance of maraviroc compared to the placebo in phase III clinical trials, its safety and its favourable pharmacological interactions profile with other drugs commonly used in HIV infected patients with comorbidity brings to light other scenarios in which Maraviroc could be useful. PMID- 19133222 TI - [Conclusions and perspectives. Maraviroc]. AB - This monograph discusses most important aspects Maraviroc, the development and application perspectives of this drug, as well as main questions raised in its use. Maraviroc is the first CCR5 antagonist approved for treating HIV infection. Its imidazopyridine structure interacts with CCR5 and induces a co-receptor conformation that prevents glycoproteins binding to the viral envelope. It has powerful antiviral activity and acts on a wide spectrum of viruses with affinity for this receptor. This situation means that a tropism test has to be done before treatment to define whether the patient is a carrier of R5 variants. Maraviroc is indicated in HIV infected patients who have received previous antiretroviral treatment. It has a low toxicity and, according to preliminary data, a high genetic barrier. The resistance mechanism is associated with changes in the V3 region which allow the virus to recognise the CCR5 co-receptor bound to the Maraviroc molecule. The main cause of treatment failure is the selection of pre existing X4 strains not detected by the reference test. Maraviroc can be combined with any other antiretroviral on the market or in clinically advanced development. The indication of Maraviroc in the early phases of the infection is not currently recommended and will depend whether it is shown to be inferior when compared other treatments or a benefit in other pathogenic aspects, such as recovery of CD4 lymphocytes or a reduction in viral reservoirs. PMID- 19133225 TI - Stimulation of EAAC1 in C6 glioma cells by store-operated calcium influx. AB - This study investigated how modulation of intracellular calcium alters the functional activity of the EAAC1 glutamate transporter in C6 glioma cells. Pre incubation of C6 glioma cells with the endoplasmic reticulum Ca2+ ATP pump inhibitor, thapsigargin (10 microM) produced a time-dependent increase in the Vmax for D-[3H]aspartate transport that reached a maximum at 15 min (143% of control; P<0.001) that was accompanied by increased plasma membrane expression of EAAC1 and was blocked by inhibition of protein kinase C. Pre-incubation of C6 glioma cells with phorbol myristate-3-acetate (100 nM for 20 min) also caused a significant increase in the Vmax of sodium-dependent D-[3H]aspartate transport (190% of control; P<0.01). In contrast, in the absence of extracellular calcium, thapsigargin caused a significant inhibition in D-[3H]aspartate transport that was not mediated by protein kinase C. Blockade of store-operated calcium channels with 2-aminoethoxydiphenyl borate (50 microM) or SKF 96365 (10 microM) caused a net inhibition of D-[3H]aspartate uptake. Co-incubation of C6 glioma cells with both thapsigargin and 2-aminoethoxydiphenyl borate (but not SKF 96365) prevented the increase in D-[3H]aspartate transport that was observed in the presence of thapsigargin alone. Furthermore, 2-aminoethoxydiphenyl borate, but not SKF 96365, reduced the increase in intracellular calcium that occurred following pre incubation of the cells with thapsigargin. It is concluded that, in C6 glioma cells, stimulation of EAAC1-mediated glutamate transport by thapsigargin is dependent on entry of calcium via the NSCC-1 subtype of store operated calcium channel and is mediated by protein kinase C. In contrast, in the absence of store operated calcium entry, thapsigargin inhibits transport. PMID- 19133223 TI - A global benchmark study using affinity-based biosensors. AB - To explore the variability in biosensor studies, 150 participants from 20 countries were given the same protein samples and asked to determine kinetic rate constants for the interaction. We chose a protein system that was amenable to analysis using different biosensor platforms as well as by users of different expertise levels. The two proteins (a 50-kDa Fab and a 60-kDa glutathione S transferase [GST] antigen) form a relatively high-affinity complex, so participants needed to optimize several experimental parameters, including ligand immobilization and regeneration conditions as well as analyte concentrations and injection/dissociation times. Although most participants collected binding responses that could be fit to yield kinetic parameters, the quality of a few data sets could have been improved by optimizing the assay design. Once these outliers were removed, the average reported affinity across the remaining panel of participants was 620 pM with a standard deviation of 980 pM. These results demonstrate that when this biosensor assay was designed and executed appropriately, the reported rate constants were consistent, and independent of which protein was immobilized and which biosensor was used. PMID- 19133224 TI - A microfluidic device for multiplexed protein detection in nano-liter volumes. AB - We describe a microfluidic immunoassay device that permits sensitive and quantitative multiplexed protein measurements on nano-liter-scale samples. The device exploits the combined power of integrated microfluidics and optically encoded microspheres to create an array of approximately 100-microm(2) sensors functionalized with capture antibodies directed against distinct targets. This strategy overcomes the need for performing biochemical coupling of affinity reagents to the device substrate, permits multiple proteins to be detected in a nano-liter-scale sample, is scalable to large numbers of samples, and has the required sensitivity to measure the abundance of proteins derived from single mammalian cells. The sensitivity of the device is sufficient to detect 1000 copies of tumor necrosis factor (TNF) in a volume of 4.7nl. PMID- 19133226 TI - Active 1918 pandemic flu viral neuraminidase has distinct N-glycan profile and is resistant to trypsin digestion. AB - The 1918 pandemic flu virus caused one of the most deadly pandemics in human history. To search for unique structural features of the neuraminidase from this virus that might have contributed to its unusual virulence, we expressed this enzyme. The purified enzyme appeared as a monomer, a dimer and a tetramer, with only the tetramer being active and therefore biologically relevant. The monomer and the dimer could not be oligomerized into the tetramer in solution, suggesting that some unique structural features were required for oligomerization and activation. These features could be related to N-glycosylation, because the tetramer displayed different N-glycans than the monomer and the dimer. Furthermore, the tetramer was found to be resistant to trypsin digestion, which may give the virus the capability to invade tissues that are normally not infected by influenza viruses and make the virus more robust for infection. PMID- 19133227 TI - Characterization of novel 2-oxoglutarate dependent dioxygenases converting L proline to cis-4-hydroxy-l-proline. AB - Hydroxyprolines are valuable chiral building blocks for organic synthesis of pharmaceuticals. Several microorganisms producing L-proline trans-4- and cis-3 hydroxylase were discovered and these enzymes were applied to the industrial production of trans-4- and cis-3-hydroxy-L-proline, respectively. Meanwhile, other hydroxyproline isomers, cis-4- and trans-3-hydroxy-L-proline, were not easily available because the corresponding hydroxylase have not been discovered. Herein we report novel L-proline cis-4-hydroxylases converting free L-proline to cis-4-hydroxy-L-proline. Two genes encoding uncharacterized proteins from Mesorhizobium loti and Sinorhizobium meliloti were cloned and overexpressed in Escherichia coli, respectively. The functions of purified proteins were investigated in detail, and consequently we detected L-proline cis-4-hydroxylase activity in both proteins. Likewise L-proline trans-4-, cis-3-hydroxylase and prolyl hydroxylase, these enzymes belonged to a 2-oxoglutarate dependent dioxygenase family and required a non-heme ferrous ion. Although their reaction mechanisms were similar to other hydroxylases, the amino acid sequence homology was not observed (less than 40%). PMID- 19133228 TI - Clustering of disease-causing mutations on the domain-domain interfaces of ABCC6. AB - Mutations in ABCC6 are responsible for pseudoxanthoma elasticum (PXE), a rare genetic disease affecting the elastic tissues of the body. ABCC6 encodes a 1503 amino acid long ABC transporter, ABCC6/MRP6. The functional link between the impaired activity of the protein and the disease is not known. We have built a homology model of this transporter, and analyzed the distribution of the known 119 missense PXE-associated mutations within the predicted structure. Significant clustering of the missense mutations has been found at complex domain-domain interfaces: at the transmission interface that involves four intracellular loops and the two ABC domains as well as at the ABC-ABC interacting surfaces. The mutations affecting these regions are 2.75 and 3.53-fold more frequent than the average mutational rate along the transporter protein sequence. These data provide a genetic proof of the importance of these domain-domain interactions in the ABCC6 transporter. PMID- 19133229 TI - Recombination rates of human microRNA. AB - The fact that microRNAs play a role in almost all biological processes is well established, as is the importance of recombination in generating genome variability. However, the association between microRNAs and recombination remains largely unknown. In order to investigate the recombination patterns of microRNAs, we performed a comprehensive analysis of the recombination rate of human microRNAs. We observed that microRNAs that are expressed in several tissues tend to have lower recombination rates than tissue-specific microRNAs. Additionally, microRNAs that are associated with a number of diseases are also likely to have lower recombination rates. Furthermore, microRNAs with higher expression levels are found to have fewer recombination events. These findings reveal patterns in recombination rates of microRNAs that could help in understanding the function, evolution, and disease-related roles of microRNAs. PMID- 19133230 TI - A role of helix 12 of the vitamin D receptor in SMRT corepressor interaction. AB - To repress gene transcription, the unliganded nuclear receptor (NR) recruits the N-CoR and SMRT corepressors via its direct association with the conserved motif within bipartite NR-interaction domains (IDs) of corepressors. We recently reported that SMRT is directly involved in the VDR-mediated repression via an ID1 specific interaction with the VDR. Here we show that removal of helix 12 from VDR (VDRDeltaAF2) converts it to a more potent repressor through additional interaction between the VDR and SMRT-ID2 in yeast and mammalian systems. These data suggest that the VDR helix 12 actively regulates the ID1 preference of the VDR by inhibiting ID2-VDR association. Using the one- plus two-hybrid system, we identified specific residues within the extended helix motif of SMRT-ID2 that are required for VDRDeltaAF2 binding. Analyses of these mutants also revealed the specific residues of SMRT-ID2 generally required for optimal NR binding as well as those involved in preferential interaction with specific NRs. PMID- 19133232 TI - A neuromedin-pyrokinin-like neuropeptide signaling system in Caenorhabditis elegans. AB - Neuromedin U (NMU) in vertebrates is a structurally highly conserved neuropeptide of which highest levels are found in the pituitary and gastrointestinal tract. In Drosophila, two neuropeptide genes encoding pyrokinins (PKs), capability (capa) and hugin, are possible insect homologs of vertebrate NMU. Here, the ligand for an orphan G protein-coupled receptor in the nematode Caenorhabditis elegans (Ce PK-R) was found using a bioinformatics approach. After cloning and expressing Ce PK-R in HEK293T cells, we found that it was activated by a neuropeptide from the C. elegans NLP-44 precursor (EC(50)=18nM). This neuropeptide precursor is reminiscent of insect CAPA precursors since it encodes a PK-like peptide and two periviscerokinin-like peptides (PVKs). Analogous to CAPA peptides in insects and NMUs in vertebrates, whole mount immunostaining in C. elegans revealed that the CAPA precursor is expressed in the nervous system. The present data also suggest that the ancestral CAPA precursor was already present in the common ancestor of Protostomians and Deuterostomians and that it might have been duplicated into CAPA and HUGIN in insects. In vertebrates, NMU is the putative homolog of a protostomian CAPA-PK. PMID- 19133231 TI - Constitutively active erythropoietin receptor expression in breast cancer cells promotes cellular proliferation and migration through a MAP-kinase dependent pathway. AB - The role of erythropoietin receptor (EpoR) expression in tumor cells and the potential of EpoR-mediated signaling to contribute to cellular proliferation and invasiveness require further characterization. To determine whether EpoR expression and activation in tumor cells modulates intracellular signal transduction to promote cellular proliferation and migration, we employed a novel experimental model using human breast cancer cells engineered to stably express a constitutively active EpoR-R129C variant. EpoR-R129C expression resulted in increased cellular proliferation and migration of breast cancer cells and these effects were associated with significantly increased Epo-induced phosphorylation of ERK1/2, AKT and c-Jun-NH2-kinase (SAPK/JNK) proteins. Expression of the constitutively active EpoR-R129C receptor promoted the proliferation and migration of breast cancer cells via activation of ERK- and SAPK/JNK-dependent signaling pathways, respectively. These findings suggest that EpoR over expression and activation in breast cancer cells has the potential to contribute to tumor progression by promoting the proliferation and invasiveness of the neoplastic cells. PMID- 19133233 TI - MSC response to pH levels found in degenerating intervertebral discs. AB - Painful degenerative disc disease is a major health problem and for successful tissue regeneration, MSCs must endure and thrive in a harsh disc microenvironment that includes matrix acidity as a critical factor. MSCs were isolated from bone marrow of Sprague-Dawley rats from two different age groups (<1 month, n=6 and 4 5 months, n=6) and cultured under four different pH conditions representative of the healthy, mildly or severely degenerated intervertebral disc (pH 7.4, 7.1, 6.8, and 6.5) for 5 days. Acidity caused an inhibition of aggrecan, collagen-1, and TIMP-3 expression, as well as a decrease in proliferation and viability and was associated with a change in cell morphology. Ageing had generally minor effects but young MSCs maintained greater mRNA expression levels. As acidic pH levels are typical of increasingly degenerated discs, our findings demonstrate the importance of early interventions and predifferentiation when planning to use MSCs for reparative treatments. PMID- 19133234 TI - Stress-induced overexpression of the heme-regulated eIF-2alpha kinase is regulated by Elk-1 activated through ERK pathway. AB - The heme-regulated eIF-2alpha kinase, also called the heme-regulated inhibitor (HRI), is activated under various cytoplasmic stresses in reticulocytes leading to inhibition of initiation of protein synthesis. Our previous studies indicated that the promoter activity and expression of the human HRI (hHRI) increase in human K562 cells during heat shock and lead exposure. Contrary to this, hemin chloride which inactivates the kinase, downregulates HRI expression. Here, we attempted to understand the mechanism of regulation of hHRI expression in the lead- and hemin-exposed cells. Our results demonstrate the involvement of two transcription factors, Elk-1 and MZF-1 in regulating HRI expression. Chromatin immunoprecipitation assays established further that Elk-1 is involved in upregulating HRI expression during stress along with a co-activator p300, while MZF-1 along with HDAC-1 is instrumental in its downregulation during hemin treatment. We also demonstrate the involvement of ERK pathway in activating Elk-1 during stress resulting in an over expression of hHRI. PMID- 19133235 TI - A novel small molecule, LAS-0811, inhibits alcohol-induced apoptosis in VL-17A cells. AB - One of the pathways by which alcohol induces hepatocyte apoptosis is via oxidative stress. We screened several chemically-synthesized small molecules and found LAS-0811, which inhibits oxidative stress. In this study, we elucidated its role in inhibiting alcohol-induced apoptosis in hepatocyte-like VL-17A cells. VL 17A cells were pre-incubated with LAS-0811, followed by ethanol incubation. Ethanol-induced reactive oxygen species and apoptosis were significantly inhibited in LAS-0811 pre-treated cells. VL-17A cells were transfected with a reporter (ARE/TK-GFP) plasmid containing green fluorescent protein (GFP) as a reporter gene and the anti-oxidant response element as the promoter. LAS-0811 pre treatment significantly induced the GFP expression compared to the cells treated with ethanol alone. LAS-0811 induced the activation of nrf2 and enhanced the expression and activity of glutathione peroxidase, one of the downstream targets of nrf2. The results indicate that LAS-0811 protects VL-17A cells against ethanol induced oxidative stress and apoptosis at least in part via nrf2 activation. PMID- 19133236 TI - Role of naloxone as an exogenous opioid receptor antagonist in spatial learning and memory of female rats during the estrous cycle. AB - Opioid neurotransmitters play a modulatory role in learning and memory processing. Their levels fluctuate throughout the reproductive cycle. The purpose of this research described herein was to study the effects of an exogenous opioid receptor antagonist, naloxone, on spatial learning and memory during the estrous cycle in female rats. Proestrus and estrus female rats were trained in a Y-maze. After a 4-hour delay, spatial recognition memory was assessed. The rats were administered naloxone (2 mg/kg) or saline before training, after training or before the retention test. The administration of naloxone to the estrus and proestrus rats before and after training had no significant effects on their preference for the novel arm of the Y-maze. Injection of naloxone to estrus rats before the retention test enhanced their preference for the novel arm of the Y maze, whereas, in the case of the proestrus rats, naloxone decreased their preference for the novel arm. Therefore, it can be concluded that naloxone enhances the retrieval of spatial recognition memory in the estrus phase rat, but that it impairs retrieval in the proestrus phase rat. This finding indicates that there is an interaction between ovarian hormone levels and opioids in cognitive function, so that naloxone prevents the facilitatory role of ovarian hormones in the retrieval of spatial memory. PMID- 19133237 TI - Hippocampal gene expression changes during age-related cognitive decline. AB - As humans age, cognitive performance decreases differentially across individuals. This age-related decline in otherwise healthy individuals is likely due to the interaction of multiple factors including genetics and environment. We hypothesized that altered spatial memory performance in genetically similar mice could be in part due to differential gene expression patterns in the hippocampus. To investigate this we utilized Morris water maze (MWM) testing in a group of young (3 months) and aged (24 months) C57BL/J male mice. Two sub-groups were identified in the aged animals; one in which MWM performance was not significantly different when compared to the young animals (aged-unimpaired; "AU") and one in which performance was significantly different by 1.5 standard deviations from the mean (aged-impaired; "AI"). One week after testing was completed the entire hippocampus was collected from six each of AU, AI and young mice and their gene expression profiles were compared using Affymetrix microarrays. Benjamini and Hochberg FDR correction at p<0.05 identified 18 genes differentially expressed between the AI and AU mice. The correlation between behavioral deficits and gene expression patterning allows a better understanding of how altered gene expression in the hippocampus contributes to accelerated age related cognitive decline and delineates between gene expression changes associated with normal aging vs. memory performance. PMID- 19133238 TI - Copper suppresses hippocampus LTP in the rat, but does not alter learning or memory in the morris water maze. AB - The objective of our study was to determinate the effect of copper on long-term potentiation (LTP) in hippocampus slices and a learning test in the Morris Water Maze (MWM). A group of adult Wistar rats received intraperitoneal (ip) injections of 1 mg/kg of CuSO(4) dissolved in saline for 30 consecutive days (Cu.R). A group of control rats (Sal.R), received saline by the same routes and duration. After this period, every individual of both groups was submitted to learning in MWM. Once the learning was completed, the LTP was studied in slices of hippocampus of both groups. The statistical assessment shows that the rats in both groups did not show significant differences in their progressive learning, notwithstanding that group Cu.R had 14.2 times more copper in their hippocampus and 16.7 times more in the visual cortices than in those of group Sal.R. On the other hand, the neurons of CA1 in hippocampus slices of Sal.R showed a significant development of LTP, but this was not observed in group Cu.R. In a second situation, 13 rats received training in MWM. Then, a group of 6 animals were injected with copper i.p. at the dose and time previously described. The 7 other animals were administered saline. Afterward, both groups were retrained in the MWM. The results obtained in Cu.R were similar to those obtained in Sal.R. Both groups maintained the concentrations of copper in the hippocampus indicated above, nonetheless, only the hippocampus slices of Cu.R did not show LTP. The spatial learning behavior of the rats was not affected by high copper concentration. PMID- 19133145 TI - Iron behaving badly: inappropriate iron chelation as a major contributor to the aetiology of vascular and other progressive inflammatory and degenerative diseases. AB - BACKGROUND: The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular 'reactive oxygen species' (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. REVIEW: We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation).The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible.This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, since in some circumstances (especially the presence of poorly liganded iron) molecules that are nominally antioxidants can actually act as pro-oxidants. The reduction of redox stress thus requires suitable levels of both antioxidants and effective iron chelators. Some polyphenolic antioxidants may serve both roles.Understanding the exact speciation and liganding of iron in all its states is thus crucial to separating its various pro- and anti-inflammatory activities. Redox stress, innate immunity and pro- (and some anti-)inflammatory cytokines are linked in particular via signalling pathways involving NF-kappaB and p38, with the oxidative roles of iron here seemingly involved upstream of the IkappaB kinase (IKK) reaction. In a number of cases it is possible to identify mechanisms by which ROSs and poorly liganded iron act synergistically and autocatalytically, leading to 'runaway' reactions that are hard to control unless one tackles multiple sites of action simultaneously. Some molecules such as statins and erythropoietin, not traditionally associated with anti-inflammatory activity, do indeed have 'pleiotropic' anti-inflammatory effects that may be of benefit here. CONCLUSION: Overall we argue, by synthesising a widely dispersed literature, that the role of poorly liganded iron has been rather underappreciated in the past, and that in combination with peroxide and superoxide its activity underpins the behaviour of a great many physiological processes that degrade over time. Understanding these requires an integrative, systems-level approach that may lead to novel therapeutic targets. PMID- 19133239 TI - Regional fMRI brain activation does correlate with global brain volume. AB - In healthy elderly participants, there has been little correlation found between measures of global brain atrophy and functional magnetic resonance imaging activated voxel counts and mean percent magnetic resonance signal change. We used a validated passive viewing task to calculate the fMRI activated voxel counts and mean percent MR signal change from the striate and ventral extrastriate cortices of 24 healthy control subjects (age range=33-89, mean age=66). We correlated these data with three measures of atrophy - percent brain volume, normalised brain volume and grey matter volume - to evaluate the relationship between global brain atrophy and regional fMRI activation. There was a strong positive correlation (r>0.90) between regional activation volume in striate and ventral extrastriate cortex and decreasing total brain volume when results were averaged by decade. Correlation with percent brain volume was particularly strong. In addition, we found a strong negative correlation between mean percent MR signal change and brain volume measures. Within-subject individual correlation was less strong, highlighting the problem of individual variability in brain volume and regional activation measures. PMID- 19133240 TI - Water-soluble Coenzyme Q10 formulation (Q-ter) promotes outer hair cell survival in a guinea pig model of noise induced hearing loss (NIHL). AB - The mitochondrial respiratory chain is a powerful source of reactive oxygen species (ROS) also in noise induced hearing loss (NIHL) and anti-oxidants and free-radicals scavengers have been shown to attenuate the damage. Coenzyme Q(10) (CoQ(10)) or ubiquinone has a bioenergetic role as a component of the mithocondrial respiratory chain, it inhibits mitochondrial lipid peroxidation, inducing ATP production and it is involved in ROS removal and prevention of oxidative stress-induced apoptosis. However the therapeutic application of CoQ(10) is limited by the lack of solubility and poor bio- availability, therefore it is a challenge to improve its water solubility in order to ameliorate the efficacy in tissues and fluids. This study was conducted in a model of acoustic trauma in the guinea pig where the effectiveness of CoQ(10) was compared with a soluble formulation of CoQ(10) (multicomposite CoQ(10) Terclatrate, Q-ter) given intraperitoneally 1 h before and once daily for 3 days after pure tone noise exposure (6 kHz for 1 h at 120 dB SPL). Functional and morphological studies were carried out by measuring auditory brainstem responses, scanning electron microscopy for hair cell loss count, active caspase 3 staining and terminal deoxynucleotidyl transferase-mediated dUTP labelling assay in order to identify initial signs of apoptosis. Treatments decreased active caspase 3 expression and the number of apoptotic cells, but animals injected with Q-ter showed a greater degree of activity in preventing apoptosis and thus in improving hearing. These data confirm that solubility of Coenzyme Q(10) improves the ability of CoQ(10) in preventing oxidative injuries that result from mitochondrial dysfunction. PMID- 19133242 TI - D-serine induces lipid and protein oxidative damage and decreases glutathione levels in brain cortex of rats. AB - The present work investigated the in vitro effects of D-serine (D-Ser) on important parameters of oxidative stress in cerebral cortex of young rats. Our results show that D-Ser significantly induced lipid peroxidation, as determined by increase of thiobarbituric acid-reactive substances and chemiluminescence levels, as well as protein oxidative damage since carbonyl formation and sulfhydryl oxidation were enhanced by this amino acid. Furthermore, the addition of free radical scavengers significantly prevented D-Ser-induced lipid oxidative damage, suggesting that free radicals were involved in this effect. D-Ser also significantly diminished glutathione levels in cortical supernatants, decreasing therefore the major brain antioxidant defense. Finally, D-Ser oxidized a glutathione commercial solution in a medium devoid of brain supernatants, indicating that it behaved as a direct acting oxidant. In contrast, L-serine, L alanine and L-threonine at concentrations as high as 5 mM did not significantly change chemiluminescence values, carbonyl content and GSH concentrations, implying a selective effect for D-serine. However, cortical supernatants exposed to 5 mM L-serine for different periods resulted in a gradual enhancement of TBA RS levels as pre-incubation time increased. The present data indicate that D-Ser induces oxidative stress in cerebral cortex of young rats. Therefore, it is presumed that this mechanism may be involved at least in part in the neurological damage found in patients affected by disorders in which D-Ser metabolism is compromised, leading to altered concentrations of this D-amino acid. PMID- 19133244 TI - Subthalamic neurons show increased firing to 5-HT2C receptor activation in 6 hydroxydopamine-lesioned rats. AB - The subthalamic nucleus is innervated by 5-HT afferents from the dorsal raphe nucleus and expresses high density of 5-HT(2C) receptors. However, the role of these receptors in neuronal firing of subthalamic neurons in vivo is unknown. In the present study, we examined the changes in the firing rate and firing pattern of subthalamic neurons, and the effect of the nonselective 5-HT(2C) receptor agonist m-CPP and selective antagonist SB242084 on the neuronal firing of subthalamic neurons in normal rats, sham rats, and rats with 6-hydroxydopamine lesions of the substantia nigra pars compacta by using extracellular recording. Results showed an increase in the percentage of subthalamic neurons exhibiting burst-firing pattern with no change in firing rate during the third week after the lesion compared to normal rats. The systemic administration of m-CPP (20-320 microg/kg, i.v.) dose-dependently increased the firing rate of subthalamic neurons, and the local application of m-CPP, 4 microg, in the subthalamic nucleus also increased the firing rate of subthalamic neurons in the lesioned rats. Similarly, at the same doses, the systemic and local administration of m-CPP induced the excitatory effects on subthalamic neurons in normal and sham rats. The excitatory effect of m-CPP was reversed by the subsequent administration of SB242084 (200 microg/kg, i.v.). These results suggest that the response of subthalamic neurons to 5-HT(2C) receptor stimulation is not altered after 6 hydroxydopamine lesions of the substantia nigra pars compacta. PMID- 19133243 TI - Long-term consequences of URB597 administration during adolescence on cannabinoid CB1 receptor binding in brain areas. AB - Despite the alarming increment in the use and abuse of cannabis preparations among young people, little is known about possible long-term consequences of targeting the endocannabinoid system during the critical developmental period of adolescence. Therefore, we aimed to analyze possible long-lasting neurobiological consequences of enhancing endocannabinoid signalling during adolescence, by means of blocking anandamide (AEA) hydrolysis. Adolescent Wistar male rats were administered an inhibitor of AEA hydrolysis, i.e. URB597 (0, 0.1 or 0.5 mg/kg/day from postnatal days 38 to 43). The expression of brain cannabinoid receptor type 1 (CB1R) was then analyzed by [(3)H]CP-55,940 auto-radiographic binding at adulthood. Repeated URB597 administration during adolescence persistently modified CB1R binding in a region-dependent manner. A long-lasting decrease of CB1R binding levels was found in caudate-putamen, nucleus accumbens, ventral tegmental area and hippocampus, while an opposite increment was observed in the locus coeruleus. Present results provide evidence for long-lasting effects of adolescent URB597 administration. Activation of endocannabinoid transmission during the still plastic phase of adolescence may have implications for the maturational end-point of the endocannabinoid system itself, which could lead to permanent alterations in neuronal brain circuits and behavioural responses. Insights into the developmental trajectories of this neuromodulatory system may help us to better understand and prevent outcomes of neonatal and adolescent cannabis exposure. PMID- 19133241 TI - Increased leukocyte ROCK activity in patients after acute ischemic stroke. AB - BACKGROUND: Rho-kinase (ROCK) is a downstream effector of Rho GTPase that is known to regulate various pathological processes important to the development of ischemic stroke, such as thrombus formation, inflammation, and vasospasm. Inhibition of ROCK leads to decreased infarct size in animal models of ischemic stroke. This study tests the hypothesis that ROCK activity increases during the acute phase of ischemic stroke. METHODS: Serial blood samples were drawn from 10 patients with acute ischemic stroke presenting within 24 h of symptom onset and with NIHSS scores >or=4. Samples were taken at 24, 48, and 72 h. Leukocyte ROCK activity was determined by immunoblotting leukocyte lysates with antibodies to the phosphorylated form of myosin-binding subunit (P-MBS) of myosin light chain phosphatase (MLCP). MBS and P-MBS contents were normalized to alpha-tubulin, and ROCK activity was expressed as the ratio of P-MBS to MBS. ROCK activities in these 10 patients were compared to baseline ROCK activities in 10 control subjects without acute illness and matched for sex, age, and number of vascular risk factors using a two-tailed Student's t-test. RESULTS: The mean NIHSS score in patients with stroke was 15.4. ROCK activity was significantly increased at 24 and 48 h in patients after acute ischemic stroke when compared to control values, with peak elevations at 48 h after stroke onset. There was no apparent correlation between ROCK activity and stroke severity based on NIHSS. CONCLUSIONS: Leukocyte ROCK activity is increased in patients after acute ischemic stroke with maximal activity occurring about 48 h after stroke onset. These findings suggest that activation of ROCK may play a role in the pathogenesis of ischemic stroke in humans. PMID- 19133245 TI - Sox11 transcription factor modulates peripheral nerve regeneration in adult mice. AB - The ability of adult peripheral sensory neurons to undergo functional and anatomical recovery following nerve injury is due in part to successful activation of transcriptional regulatory pathways. Previous in vitro evidence had suggested that the transcription factor Sox11, a HMG-domain containing protein that is highly expressed in developing sensory neurons, is an important component of this regenerative transcriptional control program. To further test the role of Sox11 in an in vivo system, we developed a new approach to specifically target small interfering RNAs (siRNAs) conjugated to the membrane permeable molecule Penetratin to injured sensory afferents. Injection of Sox11 siRNAs into the mouse saphenous nerve caused a transient knockdown of Sox11 mRNA that transiently inhibited in vivo regeneration. Electron microscopic level analysis of Sox11 RNAi injected nerves showed that regeneration of myelinated and unmyelinated axons was inhibited. Nearly all neurons in ganglia of crushed nerves that were Sox11 immunopositive showed colabeling for the stress and injury-associated activating transcription factor 3 (ATF3). In addition, treatment with Sox11 siRNAs in vitro and in vivo caused a transcriptional and translational level reduction in ATF3 expression. These anatomical and expression data support an intrinsic role for Sox11 in events that underlie successful regeneration following peripheral nerve injury. PMID- 19133247 TI - Long-term ginsenoside consumption prevents memory loss in aged SAMP8 mice by decreasing oxidative stress and up-regulating the plasticity-related proteins in hippocampus. AB - Ginsenoside, the effective component of ginseng, has been reported to have a neuron protective effect, but the preventive effect on Alzheimer's disease (AD) related memory loss and the underlying mechanisms have not been well determined. The senescence-accelerated mouse (SAM) is a useful model of AD-related memory impairment. In the present study, SAMP8 mice aged 4 months were chronically treated with ginsenoside (3 dose groups were given ginsenoside in drinking water for 7 months). The three groups were treated with ginsenoside 50, 100 and 200 mg/kg per day, respectively. Placebo-treated aged mice and young ones (4 months old) were used as controls. In addition, SAMR1 mice were used as "normal aging" control. The beneficial role of ginsenoside was manifested in the prevention of memory loss in aged SAMP8 mice. The optimal dose of ginsenoside is 100 or 200 mg/kg per day. In ginsenoside treated groups, the Abeta level markedly decreased in hippocampus and antioxidase level significantly increased in serum. In addition, the plasticity-related proteins in hippocampus significantly increased in the two ginsenoside treated groups. The plasticity-related proteins were checked in the present study including postsynaptic density-95 (PSD-95), phosphor N-methyl-D-aspartate receptor 1 (p-NMDAR1), phospho-calcium-calmodulin dependent kinase II (p-CaMKII), phospho-protein kinase A Catalyticbeta subunit (p-PKA Cbeta) and protein kinase Cgamma subunit (PKCgamma), phospho-CREB (p-CREB) and brain derived neurotrophic factor (BDNF) etc. These findings suggest that the increase of antioxidation and up-regulation of plasticity-related proteins in hippocampus may be one of the mechanisms of ginsenoside on the memory loss prevention in aged SAMP8 mice. PMID- 19133246 TI - Effects of chronic manganese exposure on working memory in non-human primates. AB - Human exposure to manganese (Mn) has been associated with a variety of cognitive deficits including learning and memory deficits. However, results from epidemiological studies have been inconsistent in describing the nature of such cognitive deficits. The present study was conducted to evaluate the effects of chronic Mn exposure on memory functioning in non-human primates and to correlate behavioral outcome with brain Mn levels in an attempt to explain outcome variability seen in prior studies. Cynomolgus macaque monkeys were trained to perform memory-related tasks (spatial working memory, non-spatial working memory, reference memory) and exposed to manganese sulfate (15-20 mg/kg/week) over an exposure period lasting 227.5+/-17.3 days. Blood manganese levels were in the upper range of levels reported for human environmental, medical or occupational exposures. By the end of the manganese exposure period, animals developed mild deficits in spatial working memory, more significant deficits in non-spatial working memory and no deficits in reference memory. Linear regression analyses showed that for most brain regions sampled, there was a significant inverse relationship between working memory task performance and brain Mn concentration. These results suggest that chronic exposure to levels of manganese achieved in this study may have detrimental effects on working memory and that Mn levels achieved in several brain regions are inversely related to working memory performance. PMID- 19133248 TI - Different reactivities of enzyme-linked immunosorbent assays for hepatocyte growth factor. AB - BACKGROUND: Hepatocyte growth factor (HGF) plays diverse roles in organ development, tissue regeneration, and tumor progression. Measurement of HGF concentrations in blood and tissues using enzyme-linked immunosorbent assay (ELISA) is a simple and easy way to understand the significance of HGF in tissue regeneration, pathophysiology, and diagnosis. METHODS: We evaluated 3 ELISA kits from different sources, referred to herein as kits A, B, and C for convenience. RESULTS: We found that the concentrations of human HGF determined using ELISA vary significantly depending on the source of the ELISA kit. Kits A and B detected both single-chain pro-HGF and 2-chain mature HGF, but kit C detected only 2-chain HGF. A difference in reactivity was also detected during analysis of plasma samples. When rat plasma collected 4 h after subcutaneous administration of human HGF was analyzed, the HGF concentration determined using kit B was remarkably higher than those obtained using kits A and C. Results of a biological assay and Western blot analysis indicated that kit B detects even degraded HGF, by which the HGF concentrations determined using kit B were significantly overestimated. CONCLUSIONS: This information serves as a guide for the selection of ELISA kits for human HGF. PMID- 19133249 TI - Upregulation and secretion of macrophage inhibitory cytokine-1 (MIC-1) in gastric cancers. AB - BACKGROUND: Macrophage inhibitory cytokine-1 (MIC-1), a distant member of the transforming growth factor (TGF)-beta superfamily, has been reported to be upregulated and secreted from several cancers. We examined MIC-1 expression and secretion in gastric cancers. METHODS: MIC-1 mRNA and protein levels in cancer tissues and cell lines were analyzed by RT-PCR and Western blot. MIC-1 expression in cancer tissues and its secretion in serum were analyzed using immunohistochemistry and ELISA. RESULTS: MIC-1 was significantly upregulated in gastric cancer tissues and cell lines. MIC-1 was secreted from gastric SNU620 cells and its levels in the serum of cancer patients were 10-fold higher than those of healthy controls. In addition, the staining of MIC-1 expression was strongly increased in metastatic gastric cancers. CONCLUSIONS: MIC-1 was obviously overexpressed in gastric cancers and MIC-1 secretion into blood may be useful for the prediction of gastric cancer progression. PMID- 19133250 TI - Determination of beta4-galactosyltransferase-7 activity using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry. AB - OBJECTIVES: Patients with Ehlers-Danlos syndrome were described to contain reduced activities of beta4-galactosyltransferase-7 (beta4Gal-T7). Therefore, measurement of beta4Gal-T7 activity can help to characterize defects in proteoglycan biosynthesis in patients with connective tissue diseases. DESIGN AND METHODS: We developed a sensitive and specific method to assay beta4Gal-T7 which is based on the transfer of galactose from UDP-galactose to the synthetic peptide Bio-BIK-F-Xyl. RESULTS: Calibration curves exhibited consistent linearity in the range of 10-2000 microg/L Bio-BIK-F-Xyl-Gal, corresponding to a beta4Gal-T7 activity of 3.5-659 microU/L. The limit of detection and the lower limit of quantification were 3.70 microg/L (1.22 microU/L) and 4.50 microg/L Bio-BIK-F-Xyl Gal (1.48 microU/L beta4Gal-T7 activity), respectively. Interassay imprecision (CV) was 8.1-13.1% in the range from 15.9 to 659 microU/L, and mean recovery was 85.3% (range 61.7-106.3%). CONCLUSIONS: This sensitive, robust and interference free LC-MS/MS assay allows an accurate determination of beta4Gal-T7 activity in human body fluids. PMID- 19133251 TI - Coenzyme Q10 distribution in blood is altered in patients with fibromyalgia. AB - OBJECTIVE: Coenzyme Q10 (CoQ(10)) is an essential electron carrier in the mitochondrial respiratory chain and a strong antioxidant. Signs and symptoms associated with muscular alteration and mitochondrial dysfunction, including oxidative stress, have been observed in patients with fibromyalgia (FM). The aim was to study CoQ(10) levels in plasma and mononuclear cells, and oxidative stress in FM patients. METHODS: We studied CoQ(10) level by HPLC in plasma and peripheral mononuclear cells obtained from patients with FM and healthy control subjects. Oxidative stress markers were analyzed in both plasma and mononuclear cells from FM patients. RESULTS: Higher level of oxidative stress markers in plasma was observed respect to control subjects. CoQ(10) level in plasma samples from FM patients was doubled compared to healthy controls and in blood mononuclear cells isolated from 37 FM patients was found to be about 40% lower. Higher levels of ROS production was observed in mononuclear cells from FM patients compared to control, and a significant decrease was induced by the presence of CoQ(10). CONCLUSION: The distribution of CoQ(10) in blood components was altered in FM patients. Also, our results confirm the oxidative stress background of this disease probably due to a defect on the distribution and metabolism of CoQ(10) in cells and tissues. The protection caused in mononuclear cells by CoQ(10) would indicate the benefit of its supplementation in FM patients. PMID- 19133252 TI - RAGE polymorphisms, renal function and histological finding at 12 months after renal transplantation. AB - OBJECTIVES: Rage (receptor for advanced glycation end products) is involved in pathogenesis of many diseases. The aim of the study was to test whether polymorphisms of RAGE gene are associated with the outcome of kidney transplantation. DESIGN AND METHODS: Four polymorphisms of the RAGE gene ( 429T/C, -374T/A, Gly82Ser and 2184A/G) were assessed in 145 renal transplant recipients and their relationship to histological changes in 12 months protocol kidney graft biopsy and renal function was examined. RESULTS: Genotype frequencies of each polymorphism corresponded to expected frequencies according to Hardy-Weinberg equilibrium. No differences between allelic and genotype frequencies among patients with normal histological findings, chronic allograft nephropathy and subclinical rejection were observed. CONCLUSION: This is the first study on polymorphisms of the RAGE gene in patients with the transplanted kidney. No association of RAGE selected gene polymorphisms with 12-months outcome of renal transplants was shown in study. PMID- 19133253 TI - Establishment of a transitory dorsal-biased window of localized Ca2+ signaling in the superficial epithelium following the mid-blastula transition in zebrafish embryos. AB - Using complementary luminescent- and fluorescent-based Ca(2+) imaging techniques, we have re-examined the Ca(2+) dynamics that occur during the Blastula Period (BP) of zebrafish development. We confirm that aperiodic, localized Ca(2+) transients are generated predominately in the superficial epithelial cells (SECs). At the start of the BP, these Ca(2+) transients are distributed homogeneously throughout the entire superficial epithelium. Following the mid blastula transition (MBT), however, their distribution becomes asymmetrical, where a significantly greater number are generated in the presumptive dorsal quadrant of the superficial epithelium. This asymmetry in Ca(2+) signaling lasts for around 60 min, after which the total number of transients generated from the entire superficial epithelium falls to less than one per minute until the end of the BP. We have thus called this asymmetry the "dorsal-biased Ca(2+) signaling window". The application of pharmacological agents indicates that the post-MBT SEC Ca(2+) transients are generated via the phosphatidylinositol (PI) signaling pathway. This suggests that the previously reported ventralizing function attributed to the homogeneously distributed PI pathway-generated SEC Ca(2+) transients is most likely to occur earlier in development, prior to the MBT. PMID- 19133254 TI - ErbB2 and ErbB3 regulate amputation-induced proliferation and migration during vertebrate regeneration. AB - Epimorphic regeneration is a unique and complex instance of postembryonic growth observed in certain metazoans that is usually triggered by severe injury [Akimenko et al., 2003; Alvarado and Tsonis, 2006; Brockes, 1997; Endo et al., 2004]. Cell division and migration are two fundamental biological processes required for supplying replacement cells during regeneration [Endo et al., 2004; Slack, 2007]. However, the connection between the early stimuli generated after injury and the signals regulating proliferation and migration during regeneration remain largely unknown. Here we show that the oncogenes ErbB2 and ErbB3, two members of the EGFR family, are essential for mounting a successful regeneration response in vertebrates. Importantly, amputation-induced progenitor proliferation and migration are significantly reduced upon genetic and/or chemical modulation of ErbB function. Moreover, we also found that NRG1 and PI3K functionally interact with ErbB2 and ErbB3 during regeneration and interfering with their function also abrogates the capacity of progenitor cells to regenerate lost structures upon amputation. Our findings suggest that ErbB, PI3K and NRG1 are components of a permissive switch for migration and proliferation continuously acting across the amputated fin from early stages of vertebrate regeneration onwards that regulate the expression of the transcription factors lef1 and msxB. PMID- 19133256 TI - MiR-101 downregulation is involved in cyclooxygenase-2 overexpression in human colon cancer cells. AB - Overexpressed cyclooxygenase-2 (COX-2) strongly contributes to the growth and invasiveness of tumoral cells in patients affected by colorectal cancer (CRC). It has been demonstrated that COX-2 overexpression depends on different cellular pathways involving both transcriptional and post-transcriptional regulations. We assumed that COX-2 expression could be regulated also by microRNAs (miRNAs) since these short RNA molecules participate to the fine regulation of several genes implicated in cell growth and differentiation. In this paper, we report the inverse correlation between COX-2 and miR-101 expression in colon cancer cell lines and we demonstrated in vitro the direct inhibition of COX-2 mRNA translation mediated by miR-101. Moreover, this correlation was supported by data collected ex vivo, in which colon cancer tissues and liver metastases derived from CRC patients were analyzed. These findings provide a novel molecular insight in the modulation of COX-2 at post-transcriptional level by miR-101 and strengthen the observation that miRNAs are highly implicated in the control of gene expression. An impairment of miR-101 levels could represent one of the leading causes of COX-2 overexpression in colon cancer cells. PMID- 19133255 TI - The selective non-peptidic delta opioid agonist SNC80 does not facilitate intracranial self-stimulation in rats. AB - Delta opioid receptor agonists are under development for a variety of clinical applications, and some findings in rats raise the possibility that agents with this mechanism have abuse liability. The present study assessed the effects of the non-peptidic delta opioid agonist SNC80 in an assay of intracranial self stimulation (ICSS) in rats. ICSS was examined at multiple stimulation frequencies to permit generation of frequency-response rate curves and evaluation of curve shifts produced by experimental manipulations. Drug-induced leftward shifts in ICSS frequency-rate curves are often interpreted as evidence of abuse liability. However, SNC80 (1.0-10 mg/kg s.c.; 10-56 mg/kg i.p.) failed to alter ICSS frequency-rate curves at doses up to those that produced convulsions in the present study or other effects (e.g. antidepressant effects) in previous studies. For comparison, the monoamine releaser d-amphetamine (0.1-1.0 mg/kg, i.p.) and the kappa agonist U69,593 (0.1-0.56 mg/kg, i.p.) produced dose-dependent leftward and rightward shifts, respectively, in ICSS frequency-rate curves, confirming the sensitivity of the procedure to drug effects. ICSS frequency-rate curves were also shifted by two non-pharmacological manipulations (reductions in stimulus intensity and increases in response requirement). Thus, SNC80 failed to facilitate or attenuate ICSS-maintained responding under conditions in which other pharmacological and non-pharmacological manipulations were effective. These results suggest that non-peptidic delta opioid receptor agonists have negligible abuse-related effects in rats. PMID- 19133257 TI - The tyrosine phosphatase SHP-2 controls urokinase-dependent signaling and functions in human vascular smooth muscle cells. AB - The urokinase (uPA)/urokinase receptor (uPAR) multifunctional system is an important mediator of functional behaviour of human vascular smooth muscle cells (VSMC). uPAR associates with platelet-derived growth factor receptor beta (PDGFR beta), which serves as a transmembrane adaptor for uPAR in VSMC, to transduce intracellular signaling and initiate functional changes. The precise and rapid propagation of these signaling cascades demands both strict and flexible regulatory mechanisms that remain unexplored. We provide evidence that the tyrosine phosphatase SHP-2 mediates these processes. uPA regulated SHP-2 phosphorylation, catalytic activity, and its co-localization and association with the PDGFR-beta. Active PDGFR-beta was required for the uPA-induced SHP-2 phosphorylation. uPAR-directed STAT1 pathway was disturbed in cells expressing SHP-2 inactive mutant. Both, cell proliferation and migration were impaired in VSMC with downregulated SHP-2. Elucidating the underlying mechanisms, we found that uPA induced SHP-2 recruitment to lipid rafts. Disruption of rafts abolished uPA-related control of SHP-2 phosphorylation, its association with PDGFR-beta and finally the VSMC functional responses. Our results demonstrate that SHP-2 plays an important role in uPA-directed signaling and functional control of human VSMC and suggest that this phosphatase might contribute to the pathogenesis of the uPA related vascular remodeling. PMID- 19133258 TI - Molecular assemblies and membrane domains in multivesicular endosome dynamics. AB - Along the degradation pathway, endosomes exhibit a characteristic multivesicular organization, resulting from the budding of vesicles into the endosomal lumen. After endocytosis and transport to early endosomes, activated signaling receptors are incorporated into these intralumenal vesicles through the action of the ESCRT machinery, a process that contributes to terminate signaling. Then, the vesicles and their protein cargo are further transported towards lysosomes for degradation. Evidence also shows that intralumenal vesicles can undergo "back fusion" with the late endosome limiting membrane, a route exploited by some pathogens and presumably followed by proteins and lipids that need to be recycled from within the endosomal lumen. This process depends on the late endosomal lipid lysobisphosphatidic acid and its putative effector Alix/AIP1, and is presumably coupled to the invagination of the endosomal limiting membrane at the molecular level via ESCRT proteins. In this review, we discuss the intra-endosomal transport routes in mammalian cells, and in particular the different mechanisms involved in membrane invagination, vesicle formation and fusion in a space inaccessible to proteins known to control intracellular membrane traffic. PMID- 19133259 TI - Collagen ultrastructural changes during stromal wound healing in organ cultured bovine corneas. AB - Corneal collagen ultrastructural changes occur during the healing process. The present study was designed to compare collagen ultrastructural changes after trephine wounding or flap creation. Bovine corneas were injured and maintained in organ culture for up to 4 weeks. Samples were removed from culture at 0, 1, 2, 3 and 4 weeks and snap frozen in liquid N(2). X-ray scattering was used to measure changes in collagen interfibrillar spacing, intermolecular spacing and fibrillar diameter. Some samples were fixed in 10% Neutral Buffered Formalin solution and wax embedded for immunohistochemistry to monitor myofibroblast differentiation in corneal flaps. Swelling effects (i.e. changes in interfibrillar spacing) were more severe in trephined corneas than in those with stromal flaps. Collagen fibrillar diameter remained normal in the periphery of injured corneas, but increased significantly in areas within and around the wound in trephined samples and in the epithelial incision site in corneal flaps. Intermolecular spacing was unchanged in all samples. In the flaps, alphaSMA expression was only detected in an area adjacent to the epithelial plug, and cell numbers gradually increased during the culture. We conclude that stromal swelling is more rapid for trephine wounded corneas than in stromal flaps, indicating that the intensity of the corneal healing response depends on the type of injury. PMID- 19133260 TI - Corneal stroma PDGF blockade and myofibroblast development. AB - Myofibroblast development and haze generation in the corneal stroma is mediated by cytokines, including transforming growth factor-beta (TGF-beta), and possibly other cytokines. This study examined the effects of stromal PDGF-beta blockade on the development of myofibroblasts in response to -9.0 diopter photorefractive keratectomy in the rabbit. Rabbits that had haze generating photorefractive keratectomy (PRK, for 9 diopters of myopia) in one eye were divided into three different groups: stromal application of plasmid pCMV.PDGFRB.23KDEL expressing a subunit of PDGF receptor b (domains 2-3, which bind PDGF-B), stromal application of empty plasmid pCMV, or stromal application of balanced salt solution (BSS). The plasmids (at a concentration 1000ng/microl) or BSS was applied to the exposed stroma immediately after surgery and every 24h for 4-5 days until the epithelium healed. The group treated with pCMV.PDGFRB.23KDEL showed lower alphaSMA+ myofibroblast density in the anterior stroma compared to either control group (P ERK1/2 involves receptor/non receptor tyrosine kinases and the Ras signaling transducer. Here we demonstrate a novel mechanism elicited by Pb for transmitting ERK1/2 signaling in CL3 human non small-cell lung adenocarcinoma cells. Pb induction of higher steady-state levels of Ras-GTP was essential for increasing phospho-Raf-1(S338) and phospho-ERK1/2. Pre-treatment of the cells with a conventional PKC inhibitor Go6976 or depleting PKCalpha using specific small interfering RNA blocked Pb induction of Ras-GTP. Pb also activated cellular tyrosine kinases. Specific pharmacological inhibitors, PD153035 for epidermal growth factor receptor (EGFR) and SU6656 for Src family tyrosine kinases (SFK), but not AG1296 for platelet-derived growth factor receptor, could suppress the Pb-induced tyrosine kinases, PKCalpha, Ras-GTP, phospho-Raf-1(S338) and phospho-ERK1/2. Furthermore, phosphorylation of tyrosines on the EGFR multiple autophosphorylation sites and the conserved SFK autophosphorylation site occurred during exposure of cells to Pb for 1-5 min and 5-30 min, respectively. Intriguingly, Pb activation of EGFR required the intrinsic kinase activity but not dimerization of the receptor. Inhibition of SFK or PKCalpha activities did not affect EGFR phosphorylation, while knockdown of EGFR blocked SFK phosphorylation and PKCalpha activation following Pb. Together, these results indicate that immediate activation of EGFR in response to Pb is obligatory for activation of SFK and PKCalpha and subsequent the Ras-Raf-1-MKK1/2 ERK1/2 signaling cascade. PMID- 19133286 TI - Data from complete mtDNA sequencing of Tunisian centenarians: testing haplogroup association and the "golden mean" to longevity. AB - Since the mitochondrial theory of ageing was proposed, mitochondrial DNA (mtDNA) diversity has been largely studied in old people, however complete genomes are still rare, being limited to Japanese and UK/US samples. In this work, we evaluated possible longevity associated polymorphisms/haplogroups in an African population, from Tunisia, by performing complete mtDNA sequencing. This population has a mixed Eurasian/sub-Saharan mtDNA gene pool, which could potentially facilitate the evaluation of association for sub-Saharan lineages. Sub-Saharan haplogroups were shown to be significantly less represented in centenarians (9.5%) than in controls (54.5%), but it is not possible to rule out an influence of population structure, which is high in these populations. No recurrent polymorphism were more frequent in centenarians than in controls, and although the Tunisian centenarians presented less synonymous and replacement polymorphisms than controls, this difference was not statistically significant. So far, it does not seem that centenarians have significantly less mildly deleterious substitutions, not only in Tunisia but also in Japanese and UK/US samples, as tested here, not favouring a "golden mean" to longevity. PMID- 19133288 TI - Stage of adoption of the human papillomavirus vaccine among college women. AB - BACKGROUND: Certain types of human papillomavirus (HPV) can cause cervical and other cancers. A vaccine that protects against HPV types responsible for 70% of cervical cancers is available to females ages 9-26. OBJECTIVE: To examine correlates of stage of vaccine adoption among women ages 18-22. METHODS: In 2007, female students (n=4774) at a New England University in the U.S. were invited to complete an on-line survey that assessed knowledge of HPV, perceived susceptibility, severity, vaccine benefits/barriers, social and subjective norms, and stage of vaccine adoption RESULTS: 1897 women (40%) responded; complete data were available for 1401. About half (53%) were planning to be vaccinated, 12% had received the vaccine, 15% were undecided, and 7% had decided against vaccination. HPV knowledge was low (mean 58%). In multivariate analyses, social norms was the strongest correlate of stage; each standard deviation increase in social norms score was associated with more than four times the odds of intending to be vaccinated within the next 30 days, compared with those who had decided against vaccination (OR=4.15; 95% CI 2.17-6.36). CONCLUSIONS: Acceptance of the vaccine was high, although misconceptions about viral transmission, availability of treatment, and the role of Pap tests were common. Perceived norms were strongly associated with intentions. Interventions on college campuses should stress vaccination as a normative behavior, provide information about viral transmission, and stress the role of continued Pap screening. PMID- 19133287 TI - Sleep deprivation increases cigarette smoking. AB - Loss of sleep may impair the ability to abstain from drug use, through any of a number of mechanisms. Sleep loss may increase drug use by impairing attention and inhibitory control, increasing the value of drug rewards over other rewards, or by inducing mood states that facilitate use of a drug. In the present study, we examined whether sleep deprivation (SD) would increase smoking in cigarette smokers, and whether it would do so by impairing attention or inhibitory control. Healthy cigarette smokers (N=14) were tested in a two-session within subject study, after overnight SD or after a normal night's sleep. Subjects were tested in both conditions in randomized order, after abstaining from cigarettes for 48 hours. The procedure was designed to model the human relapse situation. On each 6 h laboratory session after sleep or no sleep, subjects completed mood and craving questionnaires, tasks measuring behavioral inhibition and attention, and a choice procedure in which they chose between money and smoking cigarettes. SD increased self-reported fatigue and decreased arousal, it increased the number of cigarettes subjects chose to smoke, impaired behavioral inhibition and attention. However, the impairments in inhibition or attention were not related to the increase in smoking. It is possible that SD increases smoking because smokers expect that it will reduce sleepiness. Thus, the findings suggest that sleep loss may increase the likelihood of smoking during abstinence not through inhibitory or attentional mechanisms but because of the potential of nicotine to reduce subjective sleepiness. PMID- 19133289 TI - Application of immobilized horseradish peroxidase onto modified acrylonitrile copolymer membrane in removing of phenol from water. AB - A non-modified and modified with NaOH and ethylenediamine ultrafiltration membranes prepared from AN copolymer have been used as carriers for the immobilization of horseradish peroxidase (HRP) enzyme. The amount of bound protein onto the membranes and the activity of the immobilized enzyme have been investigated as well as the pH and thermal optimum, and the thermal stability of the free and immobilized HRP. The experiments have proved that the modified membrane is a better support for the immobilization of HRP enzyme. The latter has shown a greater thermal stability than the free enzyme. A possible application has been studied for reducing phenol concentration in water solutions through oxidation of phenol by hydrogen peroxide, in the presence of free and immobilized HRP enzyme on modified AN copolymer membranes. A higher degree of the phenol oxidation has been observed in the presence of the immobilized enzyme. A total removal of phenol has been achieved in the presence of immobilized HRP at concentration of the hydrogen peroxide 0.5 mmol L(-1) and concentration of the phenol in the model solutions within the interval 5-40 mg L(-1). A high degree of phenol oxidation (95.4%) has been achieved in phenol solution with 100 mg L(-1) concentration in the presence of hydrogen peroxide and immobilized HRP, which demonstrates the promising opportunity of using the enzyme for bioremediation of waste waters, containing phenol. The immobilized HRP has shown good operational stability. Deactivation of the immobilized enzyme to 50% of the initial activity has been observed after the 20th day of the enzyme operation. PMID- 19133290 TI - Surface characteristics of polyurethane elastomers based on chitin/1,4-butane diol blends. AB - Biodegradable polyurethane elastomers with tunable hydrophobicity were synthesized by step-growth polymerization techniques using poly(epsilon caprolactone) (PCL) and 4,4'-diphenylmethane diisocyanate (MDI). The prepolymer was extended with different mass ratios of chitin and 1,4-butane diol (BDO). The effect of chitin contents in chain extenders (CE) proportion on surface properties was studied and investigated. Incorporation of chitin contents into the final PU showed decrease in surface free energy and its polar component. Simultaneously, the work of water adhesion to polymer decreases significantly by increasing the chitin contents in the synthesized polymer. Contact angle measurement, water absorption and swelling behavior of the synthesized polyurethane samples were affected by varying the chitin contents in the chemical composition of the final PU. The interactions of the final PU films with solvents on the surface were displayed clear dependent on the contents of chitin in to the final polyurethane formulation. The results of different tests demonstrated that the synthesized products are a potential candidate as non-absorbable suture as previously investigated into their in vitro biocompatibility and non-toxicity [K.M. Zia, M. Zuber, I.A. Bhatti, M. Barikani, M.A. Sheikh, Int. J. Biol. Macromol. 44 (2009) 18-22]. PMID- 19133291 TI - Supramolecular organization of regenerated silkworm silk fibers. AB - The microstructures of N-methylmorpholine-N-oxide (NMMO) regenerated silk fibers have been characterized by atomic force microscopy from the micrometer to the nanometer scale and compared with those previously found from natural silks. Regenerated fibers show poor tensile properties and a brittle behavior, but their mechanical properties improve if subjected to post-spinning drawing. Consequently, it was hypothesized that post-spinning drawing would lead to a microstructure more similar to that of the natural material. Here we show that the microstructure of the samples not subjected to post-spinning drawing is composed of nanoglobules that differ from those found in natural silkworm silk both in size and orientation with respect to the macroscopic axis of the fiber. The microstructure of samples subjected to post-spinning drawing evolves in the sense of decreasing the size but increasing the orientation of the nanoglobules, but these effects are only observed in some regions of the fibers. PMID- 19133292 TI - REM sleep, energy balance and 'optimal foraging'. AB - Mechanisms underlying mammalian REM sleep (REM) indicate commonality with feeding and energy balance. REM 'epiphenomena' may facilitate this, in providing heat conservation and appetite modulation, with the atonia reflecting search (foraging?) behaviour, and the lost neck muscle tonus a suppressed ingestion. In rodents, REM deprivation severely undermines energy balance. It is argued that REM may also facilitate 'optimal foraging' in wakefulness by updating 'decisions' about: appropriate food, where to find it, allocation of time in obtaining it, the locomotion/energy expenditure required to do so, vs. risk of predation. Whereas REM in the early sleep period is oriented to this updating, later REM can put feeding 'on hold'. PGO intensity changes over successive REM periods may reflect this shift. Humans, pets, zoo and laboratory mammals, and neonates have easy food accessibility, without need to forage, leading to a potential excess of REM in the later sleep period; loss of which has different outcomes to total REM deprivation. A more ecological approach in understanding REM function is desirable. PMID- 19133293 TI - Uptake and intracellular distribution of various metal ions in human monocyte derived dendritic cells detected by Newport Green DCF diacetate ester. AB - BACKGROUND: The attempt to visualise intracellular protein metal complexes has currently been difficult due to the unavailability of probes for such molecular structures. Newport Green DCF diacetate ester is a cell permeant acetate ester, which becomes fluorescent after hydrolysis. This molecule is initially uncharged, allowing it to pass through cell membranes. Once in the cell, it is hydrolysed and becomes charged, hindering its escape from the cell and allowing it to bind charged protein metal complexes, which then become fluorescent. METHODS: In this study, we exposed cultured human monocyte-derived dendritic cells (mDC) to a variety of metal ions with the aim of having the cells take up and process protein metal complexes. Newport Green DCF diacetate ester was used to fluorescently label intracellular protein metal complexes. RESULTS: Flow cytometry analysis and confocal imaging showed specific staining for mDC exposed to aluminium, chromium, nickel, titanium and zirconium ions. The intensity of staining varied between ion types, whereby Ti(III) resulted in the brightest fluorescence signal. Aluminium, Cr(III), Ni, Ti(IV) and Zr(IV) were also clearly detectable. CONCLUSION: For the first time, intracellular metal ion protein complexes undergoing cellular processing were successfully visualised in human mDC using flow cytometry and confocal microscopy. PMID- 19133294 TI - Effects of transferrins and cytokines on nitric oxide production by an avian lymphoblastoid cell line infected with Marek's disease virus. AB - Marek's disease virus (MDV), the causative agent of Marek's disease (MD), is a herpesvirus that infects poultry causing T lymphomas. Although vaccination may prevent lymphomas formation, it is not known whether it controls viral replication and spreading in the environment. Ovotransferrin (Otrf), a member of the transferrin family, is known to exert in vitro antiviral activity in primary cultures of chicken embryo fibroblasts (CEF). In addition Otrf is produced by CEF and by an avian lymphoblastoid cell line (MDCC-MSB1) following infection/reinfection with MDV. The present work was designed to investigate the effects of reinfection and of Otrf and lactoferrin (Lf) on the production by MDCC MSB1 of nitric oxide (NO), a molecule naturally exerting an antiviral activity. These effects were also tested with two cytokines (IL-8 and IFN-gamma), alone and in association with transferrins. Synergy was found between Otrf and IFN-gamma, thus suggesting a possible role in a complementary or alternative strategy against MDV spreading. PMID- 19133295 TI - Effective ribavirin concentration in mice brain using cyclodextrin as a drug carrier: evaluation in a measles encephalitis model. AB - Ribavirin (RBV) is a water-soluble synthetic nucleoside with broad spectrum antiviral properties, but it is ineffective against major viral encephalitis because of a failure to cross the blood-brain barrier (BBB). The antiviral activity of the complex ribavirin/alpha-cyclodextrin was previously demonstrated to be stronger than free ribavirin, in an in vivo model of measles virus (MV) encephalitis in mice. The role of cyclodextrin (CD) on ribavirin uptake into the brain needs to be defined. Ribavirin specific extraction from brain tissue was developed, based on a solid phase extraction. It was quantified by high performance liquid chromatography at different time points after intraperitoneal injection of single or multiple doses of free ribavirin or of the complex ribavirin/alpha-cyclodextrin. Whatever the tested dose (40 or 100mg/kg), the amount of ribavirin in the brain was significantly higher (p<0.001) when the drug was injected as a complex with alpha-cyclodextrin, in healthy or measles virus infected mice. PMID- 19133296 TI - Exhumed from thought: basal ganglia and response learning in the plus-maze. AB - The plus-maze apparatus figured prominently in the historical debate between cognitive and stimulus-response habit learning theorists concerned with the fundamental question of "what" animals learn. An important feature of this task is that variants of the training procedure can be arranged to allow for an assessment of the relative use of cognitive/place or habit/response learning mechanisms. This brief review describes findings from several neurobiological studies published primarily over the past decade that have re-introduced the plus maze to investigate the role of the dorsal striatum in learning and memory. Converging evidence from research using brain lesion, pharmacological, and molecular/genetic approaches is described supporting the hypothesis that the dorsolateral striatum plays a selective role in response learning in the plus maze. Within a multiple systems framework of memory organization, factors that can influence the relative use of place and response learning in the plus-maze are also considered, including the nature of the visual environment, reinforcement/training parameters, and emotional state of the organism. Response learning in the plus-maze may be considered an exemplar task useful for investigating the neurobiological bases of dorsal striatal involvement in habit learning and memory. This mnemonic function of the dorsal striatum generalizes across several sensory modalities and mammalian species, including humans. PMID- 19133297 TI - Effect of an NCAM mimetic peptide FGL on impairment in spatial learning and memory after neonatal phencyclidine treatment in rats. AB - The FGL peptide is a neural cell adhesion molecule-derived fibroblast growth factor receptor agonist. FGL has both neurotrophic and memory enhancing properties. Neonatal phencyclidine (PCP) treatment on postnatal days 7, 9, and 11 has been shown to result in long-lasting behavioral abnormalities, including cognitive impairment relevant to schizophrenia. The present study investigated the effect of FGL on spatial learning and memory deficits induced by neonatal PCP treatment. Rat pups were treated with 30 mg/kg PCP on postnatal days 7, 9, and 11. Additionally, the rats were subjected to a chronic FGL treatment regimen where FGL was administered throughout development. Rats were tested as adults for spatial reference memory, reversal learning, and working memory in the Morris water maze. The PCP-treated rats demonstrated a robust impairment in working memory and reversal learning. However, the long-term memory component of the reference memory task was not affected by PCP. Chronic FGL treatment had no effect on the reversal learning impairment but ameliorated the working memory deficits almost to the levels of the control groups. In conclusion, the results suggest that the neonatal PCP treatment produced deficits in cognition relevant to schizophrenia. Moreover, working memory function was selectively protected by the neurotrophic peptide, FGL. PMID- 19133298 TI - Self-activation of Caspase-6 in vitro and in vivo: Caspase-6 activation does not induce cell death in HEK293T cells. AB - Caspase-6 (Casp6) is a short pro-domain caspase that is activated early in Alzheimer disease, yet, little is known on the mechanism of activation of this caspase. In this study, critical proteolytic processing events required for Casp6 activation in vitro and in vivo were evaluated by site directed mutagenesis of the D23 pro-domain, and D179 and D193 linker processing sites. We found that (1) Casp6 was self-processed and activated in vitro and in vivo, (2) uncleavable Casp6 possessed low activity in vitro but not in vivo, (3) the pro-domain of Casp6 entirely prevented self-processing and activation in vivo but not in vitro, (4) removal of the pro-domain promoted Casp6 activation, (5) cleavage at either D179 or D193 was sufficient to generate activity in vitro and in vivo, and (6) Casp6 activity did not induce cell death in HEK293T cells. We conclude that the Casp6 is activated through proteolytic cleavage, as are the effector Caspase-3 and -7. However, unlike other effector caspases, Casp6 can be entirely self activated and its activation does not necessarily induce cell death. PMID- 19133299 TI - The influence of calcium signaling on the regulation of alternative splicing. AB - In this review the influence of calcium signaling on the regulation of alternative splicing is discussed with respect to its influence on cell- and developmental-specific expression of different isoforms of the plasma membrane calcium pump (PMCA). In a second part the possibility is discussed that due to the interaction of the calcium-binding protein ALG-2 with a spliceosomal regulator of alternative splicing, RBM22, Ca2+-signaling may thus influence its regulatory property. PMID- 19133300 TI - Calcium fingerprints induced by calmodulin interactors in eukaryotic cells. AB - Calcium (Ca2+) is a ubiquitous second messenger which promotes cell responses through transient changes in intracellular concentrations. The prominent role of Ca2+ in cell physiology is mediated by a whole set of proteins constituting a Ca2+-signalling toolkit involved in Ca2+-signal generation, deciphering and arrest. The different Ca2+-signalosomes deliver Ca2+-signals with spatial and temporal dynamics to control the function of specific cell types. Among the intracellular proteins involved in Ca2+-signal deciphering, calmodulin (CaM) plays a pivotal role in controlling Ca2+-homeostasis and downstream Ca2+-based signalling events. Due to its ubiquitous expression in eukaryotic cells and the variety of proteins it interacts with, CaM is central in Ca2+-signalling networks. For these reasons, it is expected that disrupting or modifying CaM interactions with its target proteins will affect Ca2+-homeostasis and cellular responses. The resulting calcium response will vary depending on which interactions between CaM and target proteins are altered by the molecules and on the specific Ca2+-toolkit expressed in a given cell, even in the resting state. In the present paper, the effect of six classical CaM interactors (W5, W7, W12, W13, bifonazole and calmidazolium) was studied on Ca2+-signalling in tumor initiating cells isolated from human glioblastoma (TG1) and tobacco cells (BY-2) using the fluorescent Ca2+-sensitive Indo-1 dye and aequorin, respectively. Various Ca2+-fingerprints were obtained depending both on the CaM interactor used and the cell type investigated. These data demonstrate that interaction between the antagonists and CaM results in a differential inhibition of CaM-dependent proteins involved in Ca2+-signal regulation. In addition, the distinct Ca2+ fingerprints in tobacco and human tumor initiating glioblastoma cells induced by a given CaM interactor highlight the specificity of the Ca2+-signalosome in eukaryotic cells. PMID- 19133302 TI - Autophagic cell death: analysis in Dictyostelium. AB - Autophagic cell death (ACD) can be operationally described as cell death with an autophagic component. While most molecular bases of this autophagic component are known, in ACD the mechanism of cell death proper is not well defined, in particular because in animal cells there is poor experimental distinction between what triggers autophagy and what triggers ACD. Perhaps as a consequence, it is often thought that in animal cells a little autophagy is protective while a lot is destructive and leads to ACD, thus that the shift from autophagy to ACD is quantitative. The aim of this article is to review current knowledge on ACD in Dictyostelium, a very favorable model, with emphasis on (1) the qualitative, not quantitative nature of the shift from autophagy to ACD, in contrast to the above, and (2) random or targeted mutations of in particular the following genes: iplA (IP3R), TalB (talinB), DcsA (cellulose synthase), GbfA, ugpB, glcS (glycogen synthase) and atg1. These mutations allowed the genetic dissection of ACD features, dissociating in particular vacuolisation from cell death. PMID- 19133303 TI - Pharmacokinetics and pharmacogenomics in gastric cancer chemotherapy. AB - Despite extensive efforts, treatment of gastric cancer by chemotherapy, the globally accepted standard, is yet undetermined, and uncertainty remains regarding the optimal regimen. Recent introduction of active "new generation agents" offers hope for improving patient outcomes. Current chemotherapeutic trials provided several regimens that may become a possible standard treatment, including docetaxel/cisplatin/5-FU (TCF) and cisplatin/S-1 for advanced and metastatic cancer and S-1 monotherapy in the adjuvant setting. Along with the development of novel active regimens, individual optimization of cancer chemotherapy has been attempted in order to reduce toxicity and enhance tumor response. Unlike the rare and limited contribution of pharmacokinetic studies, pharmacogenomic studies are increasing the potential to realize the therapeutics against gastric cancer. Despite the limited data, pharmacogenomics in gastric cancer have provided a number of putative biomarkers for the prediction of tumor response to chemotherapies and of toxicity. PMID- 19133301 TI - Regulation of inositol 1,4,5-trisphosphate-induced Ca2+ release by reversible phosphorylation and dephosphorylation. AB - The inositol 1,4,5-trisphosphate (IP3) receptor (IP3R) is a universal intracellular Ca2+-release channel. It is activated after cell stimulation and plays a crucial role in the initiation and propagation of the complex spatio temporal Ca2+ signals that control cellular processes as different as fertilization, cell division, cell migration, differentiation, metabolism, muscle contraction, secretion, neuronal processing, and ultimately cell death. To achieve these various functions, often in a single cell, exquisite control of the Ca2+ release is needed. This review aims to highlight how protein kinases and protein phosphatases can interact with the IP3R or with associated proteins and so provide a large potential for fine tuning the Ca2+-release activity and for creating efficient Ca2+ signals in subcellular microdomains. PMID- 19133305 TI - Guilty or not? Feelings of guilt about food among college women. AB - The aim of this study was to assess how frequently and under what circumstances college women (n=55) experience feelings of guilt about food. Data were collected by means of a 1-week food diary. Results showed that a majority of the sample regularly experienced mild feelings of guilt. These feelings were particularly likely to result from eating between-meal snacks in the afternoon or after-dinner eating in the evening. Eating candy and ice cream accounted for the most 'guilty moments'. Eating in certain social situations (e.g., eating at a friend's house) induced relatively more often feelings of guilt. PMID- 19133306 TI - Antiviral cellular immunity in colorectal cancer patients. AB - Immunotherapy is a growing field of interest in the treatment of malignant diseases, such as colorectal cancer (CRC). The induction or enhancement of T-cell responses against tumor-associated antigens is particularly important in tumor vaccination strategies. Successful immunization relies on an intact immune system. Both chemotherapy and the tumor itself are known to potentially inhibit immune responses. In this study we analyzed T cells directed against antigens of cytomegalovirus (CMV) and influenza virus in 39 HLA-A2-positive CRC patients and 29 HLA-A2-positive healthy donors using the tetramer technology. We found no difference between CRC patients and the healthy control group for either the proportion of samples with detection of virus specific T cells or the magnitude of these specific T cells. Although we cannot draw a firm conclusion on T-cell induction in cancer patients during vaccination therapy, our results show that CRC patients retain their antiviral T cells, suggesting a potential susceptibility to immunotherapy. The quantity of adaptive immunity acquired earlier in life seems not to be affected by the presence of CRC. PMID- 19133307 TI - Utility of proteomics techniques for assessing protein expression. AB - Proteomic technologies are currently used as an effective analytical tool for examining modifications in protein profiles. Understanding the natural variation of soybean seed proteins is necessary to evaluate potential unintended (collateral) effects due to transgenic modifications in genetically modified (GMO) soybeans. We used two-dimensional polyacrylamide gel electrophoresis (2D PAGE), matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry, and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to separate, identify and quantify the different classes of soybean seed proteins. Sixteen soybean genotypes, including four wild and twelve cultivated genotypes, belonging to four different subgroups were used as models for protein profile evaluation. Significant variations of allergen and anti nutritional protein profiles were observed between two different groups, cultivated and wild soybean genotypes. However, only minor variations in protein profiles were observed within the soybean samples from the same group (cultivated or wild). These results may be useful to scientists needing to compare GMO and non-GMO soybeans once additional data are generated on additional soybean varieties and the same varieties grown at different geographical locations. PMID- 19133308 TI - Toxicity value for 3-monochloropropane-1,2-diol using a benchmark dose methodology. AB - 3-Monochloropropane-1,2-diol (alpha-chlorohydrin, 3-MCPD) is a well-known contaminant that has been detected in a wide range of foods, and that is principally generated in foods prepared by hydrochloric acid hydrolysis, such as acid-hydrolyzed vegetable protein (acid-HVP). 3-MCPD is nephrotoxic to animals at high doses and induced tumors in some organs in both sexes of rodents. NITR have recently reported on the carcinogenicity of 3-MCPD in SD rats that were exposed for 2 years to drinking water. We considered that the kidney was the main target organ for 3-MCPD in SD rats and that renal tubular hyperplasia was the most sensitive endpoint. Benchmark dose analysis of the dose-response data for renal tubular hyperplasia in male and female rats exposed to 3-MCPD in drinking water for 2 years was conducted. We applied this to the benchmark dose (BMD) methodology to yield a point of departure for developing tolerable daily intakes (TDIs). The calculated BMDs and lower-bound confidence limits (BMDLs) for the critical endpoint were estimated using the seven different models. Predicted doses associated with 10% extra risk were calculated. The smallest Akaike's Information Criterion (AIC) was used in selecting the appropriate model. The model chosen by AIC for males was the logistic and for females it was the multistage. In summary, the predicted BMD(10) and BMDL(10) were 1.21 mg/kg bw/day and 0.87 mg/kg bw/day for the male rat incidence data, and values for female rats were 26.31 mg/kg bw/day and 19.47 mg/kg bw/day. In this study, the BMDL(10) of 0.87 mg/kg bw/day for male rats was suggested as the point of departure for deriving the human tolerable daily intake level of 3-MCPD. PMID- 19133304 TI - Mucus-penetrating nanoparticles for drug and gene delivery to mucosal tissues. AB - Mucus is a viscoelastic and adhesive gel that protects the lung airways, gastrointestinal (GI) tract, vagina, eye and other mucosal surfaces. Most foreign particulates, including conventional particle-based drug delivery systems, are efficiently trapped in human mucus layers by steric obstruction and/or adhesion. Trapped particles are typically removed from the mucosal tissue within seconds to a few hours depending on anatomical location, thereby strongly limiting the duration of sustained drug delivery locally. A number of debilitating diseases could be treated more effectively and with fewer side effects if drugs and genes could be more efficiently delivered to the underlying mucosal tissues in a controlled manner. This review first describes the tenacious mucus barrier properties that have precluded the efficient penetration of therapeutic particles. It then reviews the design and development of new mucus-penetrating particles that may avoid rapid mucus clearance mechanisms, and thereby provide targeted or sustained drug delivery for localized therapies in mucosal tissues. PMID- 19133309 TI - Saponins isolated from the root of Platycodon grandiflorum protect against acute ethanol-induced hepatotoxicity in mice. AB - The protective effects of saponins isolated from the root of Platycodon grandiflorum (Changkil saponins: CKS) against alcoholic steatosis in liver injury induced by acute ethanol administration were investigated. Pretreatment with CKS prior to ethanol administration significantly prevented the increases in serum alanine aminotransferase activity, hepatic TNF-alpha level, hepatic lipid peroxidation and hepatic triglyceride level. CKS prevented ethanol-induced steatosis and necrosis, as indicated by liver histopathological studies. Additionally, CKS protected against ethanol-induced depletion of hepatic glutathione levels. CYP2E1 has been suggested as a major contributor to ethanol induced oxidative stress and liver injury. The concurrent administration of CKS efficaciously abrogated the CYP2E1 induction and CYP2E1-dependents hydroxylation of aniline as compared to the individual treatment at higher doses. These findings suggest that CKS may prevent ethanol-induced acute liver injury, possibly through its ability to block CYP2El-mediated ethanol bioactivation and its free radical scavenging effects. PMID- 19133310 TI - Possible participation of oxidative stress in causation of cell proliferation and in vivo mutagenicity in kidneys of gpt delta rats treated with potassium bromate. AB - Clarifying the participation of oxidative stress among possible contributing factors in potassium bromate (KBrO(3))-induced carcinogenesis is of importance from the perspective of human health protection. In the present study, utilizing the antioxidative effects of alpha-tocopherol (alpha-TP) or sodium ascorbic acid (SAA) to attenuate oxidative stress, alterations in bromodeoxyuridine labeling indices (BrdU-LIs) and reporter gene mutations in kidneys of male and female gpt delta rats given KBrO(3) were examined. Five male and female gpt delta rats in each group were given KBrO(3) at a concentration of 500ppm in the drinking water for 9 weeks, with 1% of alpha-TP or SAA administered in the diet from 1 week prior to the KBrO(3) treatment until the end of the experiment. Increases in 8 hydroxydeoxyguanosine levels in kidney DNA of both sexes of rats given KBrO(3) were significantly inhibited by SAA, but not alpha-TP. While BrdU-LIs in the proximal tubules of female rats were also significantly reduced by SAA, those in the males and gpt mutant frequencies in kidney DNA of both sexes were not affected by SAA or alpha-TP. Immunohistochemical and Western blot analyses for alpha(2u)-globulin strongly suggested that induction of cell proliferation observed in the males might primarily result from accumulation of this protein, independent of oxidative stress. The overall data indicated that while oxidative stress well correlates with induction of cell proliferation in females, its role in males and in generation of in vivo mutagenicity by KBrO(3) in both sexes is limited. PMID- 19133311 TI - Protective role of arjunolic acid in response to streptozotocin-induced type-I diabetes via the mitochondrial dependent and independent pathways. AB - Increasing evidences in both experimental and clinical studies suggest that oxidative stress is involved in the pathogenesis of diabetic tissue damage. Pancreatic beta-cell death is the cause of decreased insulin production in diabetes. Streptozotocin (STZ) is widely used to induce experimental diabetes due to its ability to selectively target and destroy insulin producing pancreatic beta-cells via the formation of both reactive oxygen species (ROS) and RNS (reactive nitrogen species). This study investigated the prophylactic role of arjunolic acid (AA) against STZ-induced diabetes in the pancreas tissue of the Swiss albino rats (as a working model). We observed that STZ administration (at a dose of 65mg/kg body weight, injected in the tail vain) caused increased production of both ROS and RNS in the pancreas tissue of experimental animals. Formation of these reactive intermediates decreased the intracellular antioxidant defense, increased the levels of lipid peroxidation, protein carbonylation, serum glucose and TNF-alpha. Investigating the signaling pathways, we found that STZ administration caused the activation of phospho-ERK1/2, phospho-p38, NF-kappaB and destruction of mitochondrial transmembrane potential, release of cytochrome c as well as activation of caspase 3 in the pancreas tissue keeping the levels of total ERK1/2 and p38 significantly unchanged. Treatment of animals with AA (at a dose of 20mg/kg body weight, orally) both prior and post to the STZ administration effectively reduced these adverse effects by inhibiting the excessive ROS and RNS formation as well as by down-regulating the activation of phospho-ERK1/2, phospho-p38, NF-kappaB and mitochondrial dependent signal transduction pathways leading to apoptotic cell death. Combining all, these results suggest that AA plays some beneficial roles against STZ-induced diabetes. PMID- 19133312 TI - A novel pro-apoptosis gene PNAS4 that induces apoptosis in A549 human lung adenocarcinoma cells and inhibits tumor growth in mice. AB - The gene PNAS4 is a high conservative gene that shares high homology of sequence in various organisms from plants to animals. We found overexpression of human PNAS4 induced apoptosis and arrested cell cycle in S phase in A549 human lung adenocarcinoma cells. In C57BL/6 mice model of Lewis lung carcinoma, overexpression of mouse PNAS4 significantly suppressed tumor growth and prolonged survival time through induction of tumor cell apoptosis, exhibiting effective antitumor. Our original investigations in vitro and vivo indicated PNAS4 is a novel pro-apoptosis gene, which could be used as a potential target of cancer biotherapy in future. PMID- 19133313 TI - Role of the transcriptional factor C/EBPbeta in free fatty acid-elicited beta cell failure. AB - Fatty acids can favour the development of Type 2 diabetes by reducing insulin secretion and inducing apoptosis of pancreatic beta-cells. Here, we show that sustained exposure of the beta-cell line MIN6 or of isolated pancreatic islets to the most abundant circulating fatty acid palmitate increases the level of C/EBPbeta, an insulin transcriptional repressor. In contrast, two unsaturated fatty acids, oleate and linoleate were without effect. The induction of C/EBPbeta elicited by palmitate was prevented by inhibiting the ERK1/2 MAP kinase pathway or by reducing mitochondrial fatty acid oxidation with an inhibitor of Carnitine Palmitoyl Transferase-1. Overexpression of C/EBPbeta mimicked the detrimental effects of palmitate and resulted in a drastic reduction in insulin promoter activity, impairment in the capacity to respond to secretory stimuli and an increase in apoptosis. Our data suggest a potential involvement of C/EBPbeta as mediator of the deleterious effects of unsaturated free fatty acids on beta-cell function. PMID- 19133314 TI - Expression and activity of vitamin D receptor in the human placenta and in choriocarcinoma BeWo and JEG-3 cell lines. AB - Vitamin D receptor (VDR) regulates the expression of many genes involved in mineral metabolism, cellular proliferation, differentiation and drug biotransformation. We studied the expression and activity of VDR and its heterodimerization partner retinoid X receptor-alpha (RXRalpha) in choriocarcinoma trophoblast cell lines BeWo and JEG-3, in comparison with human isolated placental cytotrophoblasts and human full term placenta. We found that VDR and RXRalpha are localised in the human term placenta trophoblast and expressed in isolated cytotrophoblasts. However, we found low expression and no transcriptional activity of VDR in used choriocarcinoma cell lines. The inhibitor of DNA methylation, 5-deoxy-3'-azacytidine, and histone deacetylase inhibitor sodium butyrate partially restored the expression of VDR, suggesting an epigenetic suppression of the gene in choriocarcinoma cells. Differentiation of BeWo cells resulted in up-regulation of VDR mRNA. Finally, we observed a non genomic effect of 1,25(OH)(2)D(3) in the activation of the extracellular signal regulated kinase (ERK) signalling pathway in JEG-3 cells. In conclusion, our results suggest an epigenetic repression of VDR gene expression and activity in choriocarcinoma cell lines, and a non-genomic effect of 1,25(OH)(2)D(3) in JEG-3 cells. PMID- 19133315 TI - Cardio-respiratory fitness, habitual physical activity and serum brain derived neurotrophic factor (BDNF) in men and women. AB - Short episodes of high intensity exercise transiently increase serum levels of BDNF in humans, but serum levels of BDNF at rest appear to be lower in more physically active humans with greater levels of energy expenditure. The relationship between serum BDNF concentration, cardio-respiratory fitness (Astrand-Rhyming test estimated VO2 max) and volume of long-term, regular exercise and sporting activity (Baecke Habitual Physical Activity Index) was investigated in 44 men and women between the age range of 18-57 years. In this group an inverse relationship between resting serum BDNF concentration and measures of both estimated VO2 max (r=-0.352; P<0.05) and long-term sporting activity (r=-0.428, P<0.01) was found. These results indicate that increased levels of cardio-respiratory fitness and habitual exercise are associated with lower resting levels of serum BDNF in healthy humans. This is the first study to demonstrate an inverse relationship between a physiological estimate of cardio respiratory fitness and serum BDNF. PMID- 19133316 TI - Platelet-derived growth factor BB promotes the migration of bone marrow-derived mesenchymal stem cells towards C6 glioma and up-regulates the expression of intracellular adhesion molecule-1. AB - Recent studies have indicated that bone marrow-derived mesenchymal stem cells (BMSCs) have the capacity of migrating towards gliomas. However, few data are available about the molecular mechanism responsible for this migratory capacity. The aim of our study was to investigate the role of platelet-derived growth factor BB (PDGFBB) in the migration of BMSCs towards C6 glioma and evaluate the effect of PDGFBB on the migrating capacity and intercellular adhesion molecule-1 (ICAM-1) expression of BMSCs. The chemokinetic activity of BMSCs in response to C6 glioma-conditioned medium and recombinant rat PDGFBB was analyzed by in vitro migration assay. The effect of PDGFBB on the expression of ICAM-1 was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence. Our data showed that C6 glioma-conditioned medium significantly increased the migration of BMSCs, which could be partially blocked by a PDGFBB neutralizing antibody. Recombinant rat PDGFBB enhanced the migration of BMSCs in a concentration-dependent way from 5 to 50ng/ml. Moreover, RT-PCR and immunofluorescence showed that 12h of 20ng/ml PDGFBB incubation could up-regulate the ICAM-1 expression of BMSCs. Our data also revealed that SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK), significantly decreased the PDGFBB-induced migration and ICAM-1 expression of BMSCs. These results demonstrate that PDGFBB contributes to the migration of BMSCs towards C6 glioma and up-regulates the expression of ICAM-1, and that p38MAPK is an important signaling molecule correlating with the signal transduction of PDGFBB-induced migration and ICAM-1 expression of BMSCs. PMID- 19133317 TI - The neuroprotective effects of NGF combined with GM1 on injured spinal cord neurons in vitro. AB - Monosialoganglioside (GM1) has been considered to have a neurotrophic factor-like activity. Nerve growth factor (NGF), a member of the neurotrophin family, is essential for neuronal survival, differentiation and maturation. The aim of the present study was to investigate whether co-administration of GM1 and NGF reverses glutamate (Glu) neurotoxicity in primary cultured rat embryonic spinal cord neurons. Spinal cord neurons were exposed to Glu (2 mmol/l), Glu (2 mmol/l) plus GM1 (10 mg/ml), Glu (2 mmol/l) plus NGF (10 ng/ml), Glu (2 mmol/l) plus GM1 (5 mg/ml) and NGF (5 ng/ml) and then processed for detecting intracellular concentrations of Ca2+([Ca2+]i) by confocal laser scanning microscopy and growth associated protein 43 (GAP43) mRNA by RT-PCR. The fluorescent intensity in Glu plus GM1 and NGF incubated neurons was the lowest as compared with that in other groups. The expression of GAP43 mRNA in Glu plus GM1 and NGF incubated neurons was the highest as compared with that in other groups. These results implicated that GM1 and NGF have synergistic neuroprotective effects on spinal cord neurons with excitotoxicity induced by Glu in vitro. PMID- 19133318 TI - Effect of olfactory bulbectomy on adenylyl cyclase activity in the limbic system. AB - Monoaminergic neurotransmission is a key element in the physiopathology of depressive disorders, but information is still sparse on animal models of this disease. Here, we used the olfactory bulbectomy (OBX) model of depression to characterize cAMP-second messenger signaling pathways, i.e., adenylyl cyclase activity (basal, sodium fluoride (NaF)- and forskolin-stimulated conditions) as well as Gi and Gs protein levels in different regions of the limbic system. Two weeks after surgery and compared to sham controls, OBX rats displayed reduced NaF stimulated adenylyl cyclase activity and increased Gi/Gs ratios in the hypothalamus, pre-frontal and cingulate cortices but not in the amygdala, hippocampus and caudate nucleus. No differences were found in basal or forskolin stimulated conditions. The observed reduction of adenylyl cyclase activity induced by NaF and the increase in the Gi/Gs ratio could explain the changes in neurotransmission in OBX rats as well as in humans with depression. PMID- 19133319 TI - Mating dominance amongst male Himalayan tahr: blonds do better. AB - In ungulates, rank order is determined by differences in weight, body size, weapon size and age. In the Caprini tribe (Bovidae: Caprinae), adult male Himalayan tahr are unique to show different coat colours, but no sexual dimorphism in weapons. A highly significant correlation between hair colour and rank order was found during the rut: males with a lighter coloured ruff dominated over darker ruffed ones, in both aggressive interactions and access to oestrus females. We studied colour-based dominance in relation to weight, age and testosterone levels, which establish the social rank in most ungulates. No differences in weight and testosterone concentrations were found between adult male colour classes, but males with paler ruffs were significantly younger than darker adult males. The distribution of physical traumas from fights confirmed that younger, lighter-coloured males had a higher rank than older, darker males, a pattern which is unusual amongst ungulates. Coat colour seems to work as a signal of rank in male-male aggressive interactions and it changes according to age, whereas the relevant physiological determinants deserve further research. Intrasexual male competition has not changed weapon size or shape in the Himalayan tahr, but ruff colours are apparently used to signal rank and dominance. Colour patterns of adult males may then be homologous to ritualised weapons, apparently being a unique feature of male tahr amongst mammals. PMID- 19133320 TI - Visual lateralization and development of spatial and social spacing behaviour of chicks (Gallus gallus domesticus). AB - We investigated whether the development of spatial behaviour of the domestic chicken is influenced by light exposure of the embryo, as is known to be the case for some other lateralized visual functions. Ninety-six chicks were incubated in the dark or exposed to light during the final days of incubation. Half of the chicks in each group had the experience of moving behind opaque screens from 10 to 12 days of age. The other half were given transparent screens as a control. Chicks were tested in a detour test and a rotated floor test and their dispersal in groups was observed in larger pens. In the rotated floor test, chicks that had had experience with opaque screens used distal cues significantly more often than chicks that had experience with transparent screens (P=0.042), regardless of whether they had been exposed to light before hatching or incubated in the dark. There were no significant differences between treatments in the detour test or in the dispersal behaviour. Hence, visual lateralization has no influence on the development of the spatial behaviour that we tested, whereas the occlusion experience is quite specific and results in shifted attention to distal spatial cues. PMID- 19133321 TI - Cone visual pigments in two species of South American marsupials. AB - Marsupials are largely confined to Australasia and to Central and South America. The visual pigments that underlie the photosensitivity of the retina have been examined in a number of species from the former group where evidence for trichromatic colour vision has been found, but none from the latter. In this paper, we report the cone opsin sequences from two nocturnal South American marsupial species, the gray short-tailed opossum, Monodelphis domestica, and the big-eared opossum, Didelphis aurita. Both are members of the Order Didelphimorphia (American opossums). For both species, only two classes of cone opsin were found, an SWS1 and an LWS sequence, and in vitro expression showed that the peak sensitivity of the SWS1 pigment is in the UV. Analysis of the Monodelphis genome confirms the absence of other classes of cone visual pigment genes. The SWS1 and LWS genes with 4 and 5 introns respectively, show the same exon-intron structure as found for these genes in all other vertebrates. The SWS1 gene shows a conserved synteny with flanking genes. The LWS gene is X-linked, as in all therian mammals so far examined, with a locus control region 1.54 kb upstream. PMID- 19133322 TI - Ethanol-induced expression of glutamate-cysteine ligase catalytic subunit gene is mediated by NF-kappaB. AB - Glutamate-cysteine ligase is a rate-limiting enzyme in the de novo synthesis of glutathione, a known scavenger of electrophiles and reactive oxygen species. Glutamate-cysteine ligase catalytic subunit (GCLC) is regulated transcriptionally by nuclear factor erythroid 2-related factor 2 (Nrf2). It has been reported that ethanol induces human GCLC production via Nrf2-mediated transactivation of the antioxidant-responsive element (ARE). Here, the luciferase reporter assay revealed the presence of an ethanol-responsive element in the human GCLC promoter; it spanned bases -1432 to -832 in hepatocytes and HepG2 cells transfected with cytochrome P450 2E1 (CYP2E1). The region lacked an ARE but had a putative nuclear factor-kappaB (NF-kappaB) element. NF-kappaB DNA-binding activity was activated in response to ethanol treatment. CYP2E1 expression was required for GCLC promoter-driven gene expression and the activation of NF kappaB. Thus ethanol-induced GCLC transcription is mediated by not only Nrf2 but also NF-kappaB. PMID- 19133323 TI - Age-related changes in choroid plexus and blood-cerebrospinal fluid barrier function in the sheep. AB - Dysfunction of the choroid plexuses (CPs) and the blood-cerebrospinal fluid barrier (BCSFB) might contribute to age-related cognitive decline and neurodegenerative disease. We used the CPs from young (1-2 years), middle-aged (3 6 years) and old (7-10 years) sheep to explore effects of aging on various aspects of CP and BCSFB functions. Total protein in the cerebrospinal fluid (CSF) was significantly higher in old compared to young sheep and CSF secretion by the CP perfused in situ was significantly lower in both old and middle-aged when compared to young sheep, which correlated with reduced (22)Na(+) uptake and efflux by the CP. Steady-state extractions of a low and medium size molecular weight extracellular space marker, (14)C-mannitol and (3)H-polyethylene glycol, respectively, were significantly higher in CPs from old compared to young animals; however, there was no significant difference in steady-state extraction of a high molecular weight marker, (125)I-bovine serum albumin. This indicates increased passive BCSFB permeability for small and medium sized molecules in old sheep. CP redox activity was significantly lower in the old animals as assessed by the MTT assay, however, there was no significant difference in ATP content and energy charge of the CP with age suggesting adequate baseline energy reserve capacity. These data indicate that normal aging processes alter protein content in the CSF, CSF secretion, integrity of the BCSFB and Na(+) flux in the epithelial layer, which could impact on CSF homeostasis and turnover. PMID- 19133324 TI - Sex differences in the relationship between obesity, C-reactive protein, physical activity, depression, sleep quality and fatigue in older adults. AB - Fatigue is a serious health concern in the elderly. Sex differences exist in adiposity, systemic inflammation, physical activity/fitness and fatigue; however, the relations among these variables remain inadequately characterized impeding the development of fatigue prevention strategies. Measures of adiposity, C reactive protein, physical activity, aerobic fitness, fatigue, sleep quality and depression were obtained from 127 community-dwelling older adults. Although similar in age (70 y) and BMI (28.0kg/m(2)) women (n=80) reported 63% greater fatigue than men (p=0.04). Adiposity (r=0.44), CRP (r=0.29), physical activity (r=-0.26) and fitness (r=-0.41) were related to fatigue in women (all p<0.05), but not in men. Depression was also related to fatigue in women (r=0.37), and was the only variable related to fatigue in men (r=0.42). In women, fatigue was independently explained (all p<0.05) by CRP (6.6%), depression (6.3%), physical activity (5.8%), and adiposity (3.9%); however, in men, only depression explained variance in fatigue (12.0%). CRP was 40% higher and adiposity 12% higher in women reporting fatigue compared to those with no fatigue; no such differences existed in men. Obese women perceived a greater degree of fatigue than non-obese women, but this was not the case in men. Women report more fatigue than men which was independently associated with inflammation, depression, physical activity and adiposity, whereas in men the only independent predictor was depression. Strategies to prevent fatigue may differ in older women and men, especially with regard to inflammation, physical activity and adiposity. PMID- 19133326 TI - Beta-cell death and proliferation after intermittent hypoxia: role of oxidative stress. AB - Intermittent hypoxia (IH), such as occurs in sleep apnea, induces increased oxidative stress and is associated with altered glucose homeostasis. Because pancreatic beta cells are very sensitive to oxidative stress we tested whether they could be affected by IH. The effects of IH exposure (24 h/day, 5.7 and 21% O(2) alternation) in mice on beta-cell proliferation and beta-cell death were tested using Ki67 staining and TUNEL staining, respectively. To assess the role of oxidative stress in these processes, transgenic mice with beta-cell-specific overexpression of the antioxidant protein MnSOD were exposed to IH. After 4 days of IH exposure, beta-cell proliferation was increased almost fourfold. Coinciding with the increase in proliferation, the subcellular localization of the cell cycle regulator cyclin D2 was increased in the nucleus. In addition, beta-cell death was increased approximately fourfold. MnSOD transgene did not alter the effects of IH on beta-cell proliferation, but completely abrogated the IH effects on cell death. Thus, IH exposure that mimics sleep apnea can lead to increased beta-cell proliferation and cell death. Furthermore, the cell death response seems to be due to oxidative stress. PMID- 19133325 TI - Fatty acid transduction of nitric oxide signaling: nitrolinoleic acid mediates protective effects through regulation of the ERK pathway. AB - In vivo and in vitro studies revealed that nitroalkenes serve as protective mediators in the lung by inducing the cytoprotective enzyme heme oxygenase-1 (HO 1). Nitrolinoleic acid (LNO2) increased HO-1 mRNA, protein, and activity in cultured pulmonary epithelial cells treated with 5 to 50 microM LNO2 and in lungs of rats injected intraperitoneally with 2.6 mg/kg LNO2 twice daily for 20 days. Western blotting revealed that HO-1 protein increased significantly within 4 h of in vitro LNO2 addition and was preceded by an increase in HO-1 mRNA, consistent with transcriptional regulation of HO-1 expression by LNO2. LNO2 also dephosphorylated and activated eukaryotic initiation factor 2alpha, a key translational regulatory protein, indicating that increased translation may also contribute to LNO2-induced increases in HO-1. Exposure of cells to LNO2 activated ERK and JNK, as evidenced by increased phosphorylation. Downstream targets of ERK and JNK, Elk-1 and c-Jun, respectively, were also phosphorylated in response to LNO2 exposure. However, inhibitor studies revealed that only the ERK pathway is necessary for the LNO2-mediated increase in HO-1 mRNA and protein. These data reveal that LNO2 induces pulmonary epithelial HO-1 expression and downstream adaptive responses to inflammation via both transcriptional and translational regulatory mechanisms. PMID- 19133328 TI - Vitamin E: the shrew waiting to be tamed. AB - Vitamin E is the last of all vitamins whose essentiality is not yet understood. Its widely accepted role as a lipophilic antioxidant has been questioned, since proof of its in vivo relevance remained scarce. The influence of vitamin E on biomarkers of oxidative stress in vivo is inconsistent and metabolites of vitamin E having reacted as an antioxidant are hardly detectable. Novel functions of vitamin E include the regulation of enzymes, most of which are membrane bound or activated by membrane recruitment. Also, expression of genes responds to vitamin E. The search for a transcription factor common to all regulated genes failed so far and a receptor that specifically binds vitamin E has not yet been identified. According to microarray data, pathways preferentially affected by the vitamin E status are the inflammatory response and cellular traffic. A role of vitamin E in cellular trafficking could best explain the neurological symptoms seen in vitamin E deficiency. Emerging knowledge on vitamin E is compiled here with the perspective to unravel the molecular mechanisms that could more likely explain the essentiality of the vitamin than its ability to scavenge free radicals. PMID- 19133327 TI - Antioxidant activity of liver growth factor, a bilirubin covalently bound to albumin. AB - We previously reported that treatment of spontaneously hypertensive rats (SHR) with liver growth factor (LGF), an albumin-bilirubin complex with a covalent bond, reduces blood pressure, improves nitric oxide (NO)-dependent vasodilatation, and exerts vascular antifibrotic actions. Because bilirubin, albumin, and albumin-bound bilirubins have antioxidant properties, we hypothesize that LGF might exert its cardiovascular actions through an antioxidant mechanism. We have tested in vitro the capacity of LGF to scavenge ABTS cation and peroxyl and hydroxyl radicals and to protect vascular NO from degradation by superoxide anion. We have also compared the antioxidant capacity of LGF with that of its molecular components albumin and bilirubin and the reference antioxidant trolox. LGF exhibited antioxidant capacity against all free radicals tested at lower concentrations than albumin, bilirubin, and trolox. LGF, bilirubin, and albumin were also able to protect endothelial NO from superoxide anion degradation in a fashion similar to that of superoxide dismutase or tiron, but at much lower concentrations. These data, together with our previous results in SHR, suggest that LGF might exert its cardiovascular regenerative actions, at least in part, through an antioxidant mechanism and that LGF could be a relevant circulating antioxidant in situations of oxidative stress. PMID- 19133330 TI - DASH diet ignored. PMID- 19133329 TI - CFTR mediates cadmium-induced apoptosis through modulation of ROS level in mouse proximal tubule cells. AB - The aim of this study was to characterize the role of CFTR during Cd(2+)-induced apoptosis. For this purpose primary cultures and cell lines originated from proximal tubules (PCT) of wild-type cftr(+/+) and cftr(-/-) mice were used. In cftr(+/+) cells, the application of Cd(2+) (5 microM) stimulated within 8 min an ERK1/2-activated CFTR-like Cl(-) conductance sensitive to CFTR(inh)-172. Thereafter Cd(2+) induced an apoptotic volume decrease (AVD) within 6 h followed by caspase-3 activation and apoptosis. The early increase in CFTR conductance was followed by the activation of volume-sensitive outwardly rectifying (VSOR) Cl(-) and TASK2 K(+) conductances. By contrast, cftr(-/-) cells exposed to Cd(2+) were unable to develop VSOR currents, caspase-3 activity, and AVD process and underwent necrosis. Moreover in cftr(+/+) cells, Cd(2+) enhanced reactive oxygen species (ROS) production and induced a 50% decrease in total glutathione content (major ROS scavenger in PCT). ROS generation and glutathione decrease depended on the presence of CFTR, since they did not occur in the presence of CFTR(inh)-172 or in cftr(-/-) cells. Additionally, Cd(2+) exposure accelerates effluxes of fluorescent glutathione S-conjugate in cftr(+/+) cells. Our data suggest that CFTR could modulate ROS levels to ensure apoptosis during Cd(2+) exposure by modulating the intracellular content of glutathione. PMID- 19133331 TI - Ask the doctor. In October 2007, I had three Cypher stents placed in my left anterior descending artery. My doctor told me to take Plavix and aspirin indefinitely. Now I need to have a hernia repaired. My surgeon refuses to operate while I'm taking Plavix and aspirin, and said I would have to stop taking the two drugs five to six days before the operation. My cardiologist says that stopping the Plavix and aspirin, as well as the stress of the operation itself, would throw me into a "hypercoagulable state," greatly increasing my chance of developing a blood clot inside one of the stents. What should I do? PMID- 19133332 TI - Ask the doctor. When I was diagnosed with atrial fibrillation four years ago, my doctor put me on a calcium-channel blocker. A year or so later, after I had angioplasty and got a stent, my doctor switched me to twice-a-day sotalol. Now I feel tired all the time and have gained weight, even though I feel like I am eating less. Can this be from the sotalol? PMID- 19133333 TI - Influence of microgroove dimension on cell behavior of human gingival fibroblasts cultured on titanium substrata. AB - OBJECTIVE: The purpose of this study was to determine the dimension of surface microgrooves on titanium (Ti) substrata that shows the greatest positive influence on characterizing specific cell behavior of cultured human gingival fibroblasts. MATERIAL AND METHODS: Commercially pure Ti disks with surface microgrooves of monotonous 3.5 mm in depth and respective 15, 30, and 60 microm in width were fabricated using photolithography and used as the culture substrata in the three experimental groups in this study (TiD15, TiD30, and TiD60 groups), whereas the smooth Ti disk was used as the control substrata (smooth Ti group). Human gingival fibroblasts were cultured on the four groups of Ti substrata on successive timelines. Cell behaviors, such as adhesion, morphology,viability and proliferation, and gene expression were analyzed and compared between all groups using crystal violet stain, scanning electron microscopy (SEM), XTT assay, and reverse transcriptase-polymerase chain reaction, respectively. RESULTS: SEM demonstrated that cells were able to readily descend into the microgrooves of TiD30 at the early phase of culture. Cells on the ridge edges or in groove corners were spindle shaped with abundant filopodia formation toward the acid etched surface inside the microgrooves, thus mimicked the cell shape in three dimensional (3D) nanoenvironment. TiD15 significantly increased the cell viability and proliferation compared with the smooth Ti substrata after 72 h of culture. Up-regulation of fibronectin(FN) and alpha5 integrin genes was noted in cells cultured on TiD15 and TiD30. Gene expression pattern specific to the cells in 3D-matrix culture, such as down-regulation of alpha-smooth muscle actin gene along with up-regulation of FN and p21 genes, was pronounced in cells cultured on TiD30. CONCLUSION: This study indicates that surface microgrooves of both 15 and 30 microm in widt hand a monotonous 3.5 microm in depth on Ti substrata increase various cell behaviors of cultured human gingival fibroblasts. PMID- 19133334 TI - Validation of a brief self-administered questionnaire for cluster headache screening in a tertiary center. AB - BACKGROUND: Cluster headache remains substantially underdiagnosed and undertreated. Early neurologic referral is indicated in patients with a suspected diagnosis of cluster headache (CH) so that management can be optimized and unnecessary procedures avoided. OBJECTIVE: To validate a brief self-administered questionnaire designed to screen CH cases in tertiary centers. METHODS: The review of clinical studies led us to identify the 3 more prevalent criteria of the second edition of the International Headache Society classification (International Classification of Headache Disorders, 2nd edition [ICHD II]) for all forms of CH (episodic and chronic forms). These 3 criteria were: strictly unilaterality of pain, attack duration <180 minutes if untreated, ipsilateral conjunctival injection, and/or lacrimation. These criteria were transformed in questions formulated in such a way that they could be self-administered and easily understood. Answer to each question was yes or no. Patients were unaided. The self-questionnaire was compared with the gold standard, the ICHD II criteria used by specialists at the university of Bordeaux headache center.We calculated the sensitivity and specificity for the 3 questions and for each pair of questions. RESULTS: The self-questionnaire was consecutively and prospectively submitted to 37 patients with CH and 59 patients with migraine. The 3-item questionnaire had a 78.4% sensibility and a 100% specificity. The 2-item questionnaire only using the attack duration associated with conjunctival injection and/or lacrimation was more sensitive (81.1%) with the same specificity (100%). CONCLUSIONS: This 2-item questionnaire could be a useful tool for screening CH cases in tertiary centers. PMID- 19133335 TI - Double checking medicines: defence against error or contributory factor? AB - RATIONALE AND AIM: The double checking of medicines in health care is a contestable procedure. It occupies an obvious position in health care practice and is understood to be an effective defence against medication error but the process is variable and the outcomes have not been exposed to testing. This paper presents an appraisal of the process using data from part of a larger study on the contributory factors in medication errors and their reporting. METHODS: Previous research studies are reviewed; data are analysed from a review of 991 drug error reports and a subsequent series of 40 in-depth interviews with health professionals in an acute hospital in northern England. RESULTS: The incident reports showed that errors occurred despite double checking but that action taken did not appear to investigate the checking process. Most interview participants (34) talked extensively about double checking but believed the process to be inconsistent. Four key categories were apparent: deference to authority, reduction of responsibility, automatic processing and lack of time. Solutions to the problems were also offered, which are discussed with several recommendations. CONCLUSIONS: Double checking medicines should be a selective and systematic procedure informed by key principles and encompassing certain behaviours. Psychological research may be instructive in reducing checking errors but the aviation industry may also have a part to play in increasing error wisdom and reducing risk. PMID- 19133336 TI - Anger over bluefin tuna decision. PMID- 19133337 TI - Acid tests. PMID- 19133338 TI - Warming worries for cod. PMID- 19133339 TI - Nearly 6.4 million Californians lacked health insurance in 2007--recession likely to reverse small gains in coverage. PMID- 19133340 TI - Low-income adolescents face more barriers to healthy weight. PMID- 19133341 TI - Commissioning. Broad sweep. PMID- 19133342 TI - Data warehouse. SUS it out. PMID- 19133343 TI - Tools and services. Magic touch. PMID- 19133344 TI - Connectivity. Info central. PMID- 19133345 TI - Skills. Knowing me, knowing you. PMID- 19133346 TI - Local to national. Primary numbers. PMID- 19133347 TI - Plan (b). AB - A raft of new rules from the IRS governing 403(b) retirement plans promises to give healthcare systems a new headache. Regular audits are part of the new system, and it means a new level of paperwork. "We're having to monitor them more, and employees are finding them less flexible. The cost to do all the recordkeeping is eating away at the employees' savings," says Sue Allyn, left, of Iowa Health System. PMID- 19133348 TI - A timely save. Citigroup bailout aids big player in municipal bonds. PMID- 19133349 TI - Cutting back. Downturn not necessarily to blame for layoffs. PMID- 19133350 TI - A good year. AHIP profits soar, but below '05 level. PMID- 19133351 TI - Time for sacrifice. With proposed deal on coverage, insurers say they'll do right -if they're paid. PMID- 19133352 TI - Auction-rate endgame. Restructuring debt has cost hospitals millions. PMID- 19133353 TI - Leadership and successful financial performance in healthcare. PMID- 19133355 TI - [Baden Wurttemberg family physician's contract: how to keep the patients in the boat?]. PMID- 19133354 TI - Inspired leadership: Dr. Patricia Gabow talks about pursuing transformation and why mentoring matters. PMID- 19133356 TI - [Treatable tumor but death through infection -- that must not happen! Complications to be diagnosed and treated on time]. PMID- 19133357 TI - [Clinical trials go on: will vaccination against Alzheimer's come soon?]. PMID- 19133358 TI - [Case report: hypokalemia as a cause of paraplegia]. PMID- 19133359 TI - [Specific program for smoking cessation: thus your patients become nonsmokers]. PMID- 19133360 TI - [Which further diagnosis is meaningful? Urine test strips: erythrocyte positive]. PMID- 19133361 TI - [Proton pump inhibition: more than only reflux therapy]. PMID- 19133362 TI - [Sign of the times]. PMID- 19133363 TI - [Decision against Duisburg family physicians. Cooperation with hospital prohibited]. PMID- 19133364 TI - [Statin cuts cardiovascular risk in half]. PMID- 19133365 TI - [Numbers only for national health insurance physicians]. PMID- 19133366 TI - [How benign is syncope?]. PMID- 19133367 TI - [Gastrointestinal toxicity of oncological agents]. PMID- 19133368 TI - [EGFR (epidermal growth factor receptor) inhibitor-associated skin disorders in tumor therapy]. PMID- 19133369 TI - [Acute urinary retention]. PMID- 19133370 TI - [Diagnosis and therapy of Alzheimer's disease: what's important for the family doctor?]. PMID- 19133371 TI - [Valsartan improves symptoms and quality of life in patients with chronic heart failure]. PMID- 19133372 TI - [Insulin therapy in type 2 diabetes. Insured patients would be willing to pay for added benefits]. PMID- 19133373 TI - Teamed up for reform. As Obama looks to teamed up solve healthcare's problems, Daschle, Lambrew healthcare team makes sense, industry executives say. AB - The president-elect sent strong signals that he's serious about reform, with some key appointments last week. The move drew applause from industry executives, including one who said the system is now set on a crash course. "There's no guarantee that someone is going to be able to put Humpty Dumpty back together, but we're never going to know until someone starts sorting through the pieces," says William Atkinson, left, of WakeMed. PMID- 19133374 TI - Ill. governor's healthy thoughts. Healthcare at center of Blagojevich allegations. PMID- 19133375 TI - Safety net frayed. With budget deficits totaling $140 billion, 43 states eyeing cuts: report. PMID- 19133376 TI - 5 million lives? ... not quite. Critics say clear standards needed to assess efficacy. PMID- 19133377 TI - HIT budgets taking a hit: study. Majority deferring purchases, extending timelines. PMID- 19133378 TI - Edging closer. Cash erodes, weak hospitals risk defaults. PMID- 19133379 TI - A little off the top. Executives urged to take pay cuts, freeze salaries. PMID- 19133380 TI - Joining reform push. Hospital exec group wants to work with lawmakers. PMID- 19133381 TI - Open the curtain on PQRI: docs. CMS told program needs to be more transparent. PMID- 19133382 TI - It's never too early. Excellence in education is key, whether it's grammar school or medical school. PMID- 19133383 TI - Patients before profits. A doc-owned hospital would raise costs, utilization. PMID- 19133384 TI - Click for savings. Web tutorials set realistic patient expectations. PMID- 19133386 TI - Spirit of Excellence Award for Service--honorable mention. Turning frowns upside down. PMID- 19133385 TI - Spirit of Excellence Award for Service--winner A side order of sensitivity. PMID- 19133387 TI - Spirit of Excellence Award for Quality--winner. Taking responsibility. PMID- 19133388 TI - Spirit of Excellence Award for Community--honorable mention. Battling injuries. PMID- 19133389 TI - Spirit of Excellence Award for Team--winner. Stopping the bleeding. PMID- 19133390 TI - Spirit of Excellence Award for Team--honorable mention. Something to talk about. PMID- 19133391 TI - Spirit of Excellence Award for C.A.R.E.S.--winner. Eating troubles addressed. PMID- 19133392 TI - Spirit of Excellence Award for C.A.R.E.S.--honorable mention. Employee outreach. PMID- 19133393 TI - Beliefwatch heavenly. PMID- 19133394 TI - For safer baby bottles. PMID- 19133395 TI - Hospital hygiene. PMID- 19133396 TI - Why the new WHO growth charts are dangerous to breastfeeding. PMID- 19133397 TI - The new WHO Child Growth Standards: possible effects on exclusive breastfeeding in the first six months. PMID- 19133398 TI - The WHO Child Growth Standards and current Western growth references. PMID- 19133399 TI - The benefits of phone support and home visits: an evaluation of the City of Kingston's Breastfeeding Support Service. AB - This paper presents an evaluation of the City of Kingston Breastfeeding Support Service that was performed in 2006. The evaluation utilised a mail-out questionnaire designed to investigate mothers' breastfeeding expectations, experiences, issues and support received, in addition to their experience of using the Service. Seventy-seven percent of mothers contacted the service for support due primarily to problems with positioning and attachment, nipple pain or mastitis. Other reasons were also cited, reflecting that it takes time and experience for women to learn to breastfeed and to develop an understanding of their lactation. The study found that on discharge from the hospital, 59% of mothers were fully breastfeeding; this figure decreased to 32% at the time of contacting the service, but increased to 42% two weeks after contacting the service. Satisfaction with both the model of service and the support received was very high. Mothers were able to feel more comfortable with their breastfeeding (75%), to breastfeed for longer (59%) and with more confidence (71%) and greater knowledge (68%) about breastfeeding and lactation. The majority of mothers (96%) reported they would contact the service again if they needed further breastfeeding advice and support. PMID- 19133400 TI - Infant feeding and professional advice in the first half of the 20th century in Greece. AB - This study aims to assess the role that health professional and State policies played in shaping breastfeeding practices and attitudes in Greece during the first half of the 20th century. Original texts were used; including those concerned with breastfeeding traditions, health professionals' attitudes to breastfeeding, infant feeding patterns, partial breastfeeding, artificial feeding and State policies for the promotion of breastfeeding. Content analysis was used and breastfeeding rates were considered. In the first two decades of the 20th century, most Greek women breastfed their children, as advised by other experienced women. In the succeeding decades, health professionals and policy makers wrote books and articles praising breastfeeding albeit stressing the nursing mothers' ignorance of sanitary measures. Many health professionals were influenced by trends in developed countries and advocated novel infant feeding practices. Consequently, full breastfeeding was not promoted. PMID- 19133401 TI - Current world literature. Protein, amino acid metabolism and therapy. PMID- 19133402 TI - Commentaries on leadership and successful financial performance in healthcare. Creating unified, inspired levels of greater accountability and leadership. PMID- 19133403 TI - Bibliography. Current world literature. Circulation and hemodynamics. PMID- 19133404 TI - Consequences of alcohol consumption on neurotransmitters -an overview. AB - Alcohol one of the important products of the global addiction alters brain function by interacting with multiple neurotransmitter systems, thereby disrupting the delicate balance between inhibitory and excitatory neurotransmitters. Alcohol positively reinforces drinking by producing a mild euphoria. The reinforcing effects of alcohol are mediated by several neurochemical systems and are associated with some of the behavioral manifestations of intoxication. Alcohol consumption is initially accompanied by decreased attention, alterations in memory, mood changes and drowsiness. Generally all vital functions of brain depend on a delicate balance between excitatory and inhibitory neurotransmission,which in turn dependent on short and long term alcohol consumption. Detailed understanding of alcohol's mechanism of action on the neurotransmitters of brain is a prerequisite in discovering effective treatments for both alcohol abuse and alcoholism. This review covers the elaborate literature on the subject and highlights the functions and interactions of neurotransmitters and alcoholism. PMID- 19133405 TI - Sphenoid mucocele after radiation for the treatment of nasopharyngeal carcinoma. PMID- 19133406 TI - Metabolic abnormalities in chronic kidney disease that contribute to cardiovascular disease, and nutritional initiatives that may diminish the risk. AB - PURPOSE OF REVIEW: Chronic kidney disease (CKD) is associated with a wide range of severe metabolic and nutritional disturbances that directly or indirectly contribute to left ventricular hypertrophy, ischemic heart disease, vascular calcification, heart failure and other manifestations of cardiovascular disease (CVD). The CVD mortality rate in CKD patients is far higher than in the general population, and CKD is today recognized as one of the most important risk factors for CVD. In this review, we discuss the links between metabolic abnormalities and CVD in CKD patients and nutritional initiatives that may reduce this risk. RECENT FINDINGS: Certain nontraditional risk factors, such as protein-energy wasting, inflammation, and biomarkers reflecting bone and mineral disorders, are strong predictors of CVD mortality in CKD patients. Although several small nutritional intervention studies have been performed and nutritional guidelines have been introduced in order to minimize metabolic disorders and improve nutritional status, they have so far not been proven to reduce morbidity nor mortality. SUMMARY: Although the pathophysiological mechanisms involved in the markedly increased CVD risk of CKD patients are becoming more evident, still few nutritional randomized controlled studies have been conducted in this high-risk patient group. PMID- 19133407 TI - Direct antiatherosclerotic effects of PPAR agonists. AB - PURPOSE OF REVIEW: Peroxisome proliferator activated receptors (PPARs) are ligand dependent transcription factors that mediate a range of important metabolic functions by transactivation, transrepression or corepression of various gene targets. PPAR agonists also have direct antiatherosclerotic effects, independent of their metabolic effects on glucose and lipid homeostasis. The purpose of this review is to evaluate the currently available evidence for a direct vasculoprotective effect of PPAR agonists. RECENT FINDINGS: Current studies have emphasized PPAR-mediated effects on inflammatory and immune responses, oxidative stress, the renin-angiotensin system and modulation of plaque composition. Furthermore, it has become evident that the relative activation of the different PPAR isoforms and the contribution of transactivation of target genes against transrepression of transcription factors need to be considered when assessing the vasculoprotective effects of PPAR agonists. SUMMARY: It is anticipated that the antiatherosclerotic effects of PPAR agonists observed in experimental studies will translate into reduced cardiovascular events. This promise is yet to be realized in short-to-medium term studies. Given the central role of the PPAR in gene regulation, particularly in metabolic states, it is possible that more targeted modulation of PPAR signalling may hold many rewards for the prevention of atherosclerosis. PMID- 19133408 TI - Inflammatory pseudotumours of the larynx: three cases and a review of the literature. PMID- 19133409 TI - Potential impact of omega-3 treatment on cardiovascular disease in type 2 diabetes. AB - PURPOSE OF REVIEW: Hypothesis-generating systematic review of the impact of marine-derived omega-3 polyunsaturated fatty acids (PUFAs) on lipid, glycemic and hematological risk factors in type 2 diabetes using pooled data from randomized controlled trials searched up to 20 September 2008. RECENT FINDINGS: Seven new trials in 2007 and 2008 were identified from 206 abstracts to give a total of 24 trials between 1966 and 2008 involving 1533 participants that could be pooled. The mean omega-3 PUFAs dose and duration of treatment in the new trials was 2.4 g/day and 24 weeks, respectively. Compared with placebo, omega-3 PUFAs supplementation decreased triglycerides by 7% (mean -0.17 mmol/l, 24 trials, 1530 participants), fibrinogen by 10% (mean -0.96 micromol/l, three trials, 159 participants), ADP platelet aggregation to ADP by 22% (mean -10.30%, two trials, 64 participants) and to collagen by 21% (mean -10.55%, two trials, 64 participants), with an LDL-cholesterol increase of 3% (mean 0.08 mmol/l, 21 trials, 1104 participants). None of the following risk factors appeared to be beneficially influenced: HDL-cholesterol, LDL particle size, glycemia, insulinemia, inflammatory biomarkers, blood pressure. However for some of these risk factors (such as inflammatory biomarkers) the number of trial patients was small Higher doses of omega-3 PUFAs (>or=2 g/day) may have greater triglyceride lowering effects. SUMMARY: This systematic review and meta-analysis confirms the triglyceride lowering effects of omega-3 PUFAs, demonstrates potential dose response effects and shows improvements in thrombogenesis. Omega-3 PUFAs raise LDL levels without concomitant changes in lipid particle size. Changes seen in conventional risk factors are insufficient to explain the cardiovascular disease risk reductions suggested to occur with omega-3 PUFAs. PMID- 19133410 TI - The effect of dietary cholesterol on macrophage accumulation in adipose tissue: implications for systemic inflammation and atherosclerosis. AB - PURPOSE OF REVIEW: It is well recognized that adipose tissue in obesity is characterized by macrophage accumulation and local inflammation. This review summarizes current evidence regarding dietary cholesterol on adipose tissue macrophage accrual, systemic inflammation and its potential link to atherosclerosis. RECENT FINDINGS: Based upon epidemiological data and animal studies, both obesity and dietary cholesterol have been associated with coronary artery disease. However, the effect of dietary cholesterol on adipose tissue has not been widely studied. In an animal model of obesity/metabolic syndrome, feeding a diabetogenic diet high in saturated fat and refined carbohydrate with 0.15% cholesterol added resulted in increased adipose tissue macrophage accumulation, local inflammation and chronic systemic inflammation compared to animals that received the same diet without added cholesterol. There also was an increased macrophage content of atherosclerotic lesions observed in the added cholesterol group. SUMMARY: Mechanisms involved in adipose tissue macrophage accrual continue to be elusive. There are limited data that dietary cholesterol may worsen macrophage accumulation in adipose tissue and the artery wall. Cytokines produced by inflamed adipose tissue may lead to inflammatory changes in the liver, which could then play a role in atherogenesis. PMID- 19133411 TI - Dietary fat oxidation as a function of body fat. AB - PURPOSE OF REVIEW: Is trafficking of dietary fat involved in the development of obesity? RECENT FINDINGS: Studies on energy expenditure and substrate utilization during overfeeding, studies on individual differences in substrate utilization between individuals fed at energy balance, and differences in responses in energy expenditure and substrate utilization in individuals after weight reduction, illustrate that the capacity of fat oxidation is a potential determinant for the development of obesity. SUMMARY: The ability to store dietary fat seems to be involved in the susceptibility to gain weight during a positive energy balance. Obese individuals show less oxidation and more storage of dietary fat as compared with the lean phenotype. Differences in fuel trafficking make individuals prone to overeating in the current obesogenic environment with a high availability of energy-dense fatty foods. It is difficult to get rid of excess body fat as energy requirement for weight maintenance after weight reduction is lower than predicted from the new body composition reached and thus, there is a high risk for weight regain. PMID- 19133412 TI - Ectopic lipids and organ function. AB - PURPOSE OF REVIEW: To summarize recent studies that shed more light on possible mechanisms by which ectopic lipid storage affects organ function. RECENT FINDINGS: Although ectopic lipids have been considered as biomarkers of lipotoxicity, adaptation of metabolic fluxes and of mitochondrial function seem to be more important than actual cellular fat contents in liver and muscle. Diabetic and obese humans have elevated myocardial lipid contents, which are associated with mitochondrial and contractile dysfunction and could even precede the development of heart failure. Although pancreatic fat content is negatively associated with insulin secretion, [beta]-cell triglycerides are not easily accessible to measurement in humans rendering their role for [beta]-cell function unclear. New approaches to quantify energy metabolism in various organs could help to identify novel biomarkers of organ function in humans. SUMMARY: Dietary intake of high-caloric high-fat diets and sedentary lifestyle lead to increased storage of triglycerides not only in adipose tissue but also ectopically in other tissues. Intracellular lipid contents in skeletal muscle and liver have been related to insulin resistance and inflammatory processes. Myocardial fat is increased in heart failure, whereas pancreatic fat could relate to insulin secretion. PMID- 19133413 TI - Re: de Souza RM, de Menezes LM. Nickel, chromium and iron levels in the saliva of patients with simulated fixed orthodontic appliances. Angle Orthod. 2008;78:345 350. PMID- 19133414 TI - New year, new collaboration. 2009 brings new partnerships in advancing HIM issues. PMID- 19133415 TI - 2008 FORE Triumph Awards. AHIMA honors those who make a difference. PMID- 19133416 TI - Professional practice solutions. Releasing records from other providers. PMID- 19133417 TI - Professional practice solutions. CMS bans use of physician signature stamps. PMID- 19133418 TI - Health hazards of aflatoxin. PMID- 19133419 TI - Aflatoxin levels in locally grown maize from Makueni District, Kenya. AB - OBJECTIVES: Investigations were carried out to determine aflatoxin levels in household maize in Makueni District and to correlate aflatoxin levels to maize drying and storage practices. Also, aflatoxin exposure in villages that reported aflatoxicosis cases in 2005 was compared with that in villages that did not report cases to assess whether aflatoxin exposure levels could be used to identify high-risk villages for targeted prevention interventions. DESIGN: A cross-sectional study. SETTING: Three divisions of Makueni district, Kibwezi, Makindu and Mtito Andei in Eastern Province, Kenya. SUBJECTS: Ninety six households were surveyed, and 104 maize samples were analysed for total aflatoxin levels from June to July 2005. The households were selected from high and low aflatoxicosis risk areas. RESULTS: Out of the 104 maize samples collected from 96 households, 37 (35.5%) had aflatoxin levels above the World Health Organisation (WHO) recommended maximum limit of 20 ppb. All of these samples were homegrown or purchased. Twenty one samples (20.1%) had levels above 100 ppb. Eleven (10.6%) had extremely high levels above 1000 ppb. No relief supply maize had aflatoxin levels above the WHO maximum limit. CONCLUSION: High levels of aflatoxin in homegrown and purchased maize suggested that aflatoxin exposure was widespread. PMID- 19133420 TI - Prevalence of hepatitis C virus and its genotypes among a cohort of drug users in Kenya. AB - BACKGROUND: Prevalence of hepatitis C virus and that of its main genotypes varies between the worlds geographic regions. The risk factors for infection with HCV include blood transfusion, tattoing and injecting drug use. OBJECTIVES: To examine the prevalence of HCV and determine its main genotypes among a cohort of drug users in Kenya. DESIGN: A laboratory based study. SETTING: Hepatitis research laboratory in the Centre for Virus Research at the Kenya Medical Research Institute, Nairobi. SUBJECTS: Three hundred and fourteen male and 19 female intravenous and non-intravenous drug users aged between 15-55 years. RESULTS: Seventy four (22.2%) out of 333 samples tested positive for anti-HCV. Sixty nine out of the 74 serum samples were assayed for HCV RNA and 38 (55.5%) were positive. The RNA positive samples were further subjected to sequencing and 19 (73%) of the samples were classified as genotype 1a, while seven (27%) samples were classified as genotype 4. Genotypes 2, 3, 5 and 6 were not identified in this study. CONCLUSIONS: These results demonstrate a high HCV infection prevalence among this cohort of drug users (22.2%) as compared to that of the general population, which is estimated to be 0.2-0.9%. The study also confirms the presence of at least two major genotypes among Kenyan drug users (genotypes 1 and 4). PMID- 19133422 TI - Modified ureterosigmoidostomy for management of malignant and non-malignant conditions. AB - OBJECTIVE: To investigate the outcome of Mainz Pouch II urinary diversion for both malignant and non-malignant diseases. DESIGN: A retrospective analysis. SETTING: Kilimanjaro Christian Medical Centre, Institute of Urology, Moshi, Tanzania from April 1995 to May 2007. PATIENTS: Mainz Pouch II was created in 83 patients of which, 38 were females and 45 were males (M:F 1.2:1). RESULTS: Early complications were seen in 11 (13.2%) patients, as follows: one (1.2%) prolonged ileus, 1(1.2%) wound dehiscence, two (2.4%) perioperative deaths among the malignant group, seven (8.4%) superficial wound sepsis. Long term complications were seen in 14 (16.9%) patients, as follows: one (1.2%) patient developed an incision hernia, one (1.2%) patient developed unilateral pyelonephritis, one (1.2%) patient developed unilateral ureteral stenosis, two (2.4%) patients had deterioration of renal function, three (3.6%) patients developed mild to moderate unilateral hydronephrosis, three (3.6%) patients developed mucoceles. Among the 83 patients in this series, three (3.6%) patients developed metabolic acidosis, two (2.4%) of which, required oral bicarbonate supplementation. All (100%) patients had daytime continence while three (3.6%) patients had occasional night time incontinence. Overall total continence was achieved in 80 (96.4%) of the patients. CONCLUSION: The Mainz Pouch II is a safe and reproducible method of urinary diversion and serves as a satisfying method of continent urinary diversion in all age groups. This reconstructive surgery enabled the afflicted to achieve personal goals, hopes and aspirations, positively influencing their quality of life. The follow up show low complication rate with good results in terms of continence and quality of life, however, long term results remain to be evaluated. PMID- 19133421 TI - Stakeholders perception of HIV sero-discordant couples in western Kenya. AB - OBJECTIVE: To describe the perceptions of key stakeholders regarding the counselling needs of HIV sero-discordant couples as part of preparation for a clinical trial involving HIV sero-discordant couples. DESIGN: Qualitative study using key informant and couple interviews. SETTING: Moi Teaching and Referral Hospital (MTRH). SUBJECTS: A purposive sample of nine key informants and 31 couple interviews totaling 71 participants. The couple interviews consisted of HIV untested, HIV concordant (positive and negative) and discordant couples. RESULTS: Seventy one individuals participated in nine key informant and 31 couple interviews. The responses identified the following as key issues in counselling HIV discordant couples: The need for education on the meaning of HIV sero discordancy including potential sources of infection; assistance in disclosing HIV test results to one's partner; discussion of the stigma surrounding formula feeding. Overall, the participants supported safer sexual practices in discordant partnerships. CONCLUSIONS: Psychosocial support of HIV sero-discordant couples should include messages about the meaning, mechanisms and implications of sero discordancy. Culturally appropriate HIV-disclosure and safer sex messages are also needed to support these partnerships. PMID- 19133423 TI - Fertility rate of epileptic women at Kenyatta National Hospital. AB - BACKGROUND: Reproductive health issues of women with epilepsy (WWE) are complex and multifaceted, and both epilepsy and antiepileptic drug (AEDs) use may alter fertility and pregnancy outcomes in these women. OBJECTIVE: To determine the fertility rate of women with epilepsy at Kenyatta National Hospital (KNH). DESIGN: Cross-sectional study. SETTING: Neurology clinic, KNH, Nairobi, Kenya, between October 2006 and March 2007. SUBJECTS: A total of 191 women with epilepsy (aged 15-49 years) who had been epileptic for at least one year were interviewed regarding their pregnancy and birth histories, and the information validated with medical records where available. RESULTS: The general fertility rate (GFR) for reproductive-aged epileptic women for the three-year period (2003-2006) preceding the study was 46 livebirths per 1000 women-years (95% CI 35.13-63.59). CONCLUSION: Fertility rate in epileptic women is decreased by two thirds (compared to that of general population of women in Kenya). Reasons for this are probably miltifactorial. PMID- 19133424 TI - Gender-associated violence at a women's hospital in Nairobi, Kenya. AB - OBJECTIVE: To describe the pattern of gender-associated violence amongst adult survivors. DESIGN: Descriptive case analysis of prospectively collected data. SETTING: The gender violence and recovery centre (GVRC), a shelter and violence treatment facility for the gender-associated violence at the Nairobi Women's hospital (NWH). SUBJECTS: Data on 663 consecutive adult patients who presented to GVRC between February 2003 and April 2004 were evaluated. RESULTS: The patients age ranged from 18 to 74 years (mean 27.7 years). Four hundred and eight patients (61.5%) presented following sexual assault. Most of assaults were perpetrated at night. A stranger was the assault perpetrator in 75.1% and 2% of sexual and non sexual assault respectively. An intimate partner was the perpetrator in majority (86.5%) of nonsexual violence. Most physical injuries were minor bruises and swellings. The rate of positive spermatozoa from high vaginal swabs was only 15.9% in cases of sexual assault. Sexual assault was significantly associated with single status of the victim and assault by more than one assailant. CONCLUSIONS: Violence against women is a common public health problem in the city of Nairobi. Women are vulnerable both in and out of the home. More efforts, including massive public education, are needed to protect this vulnerable population. PMID- 19133425 TI - Re-usable low density polyethylene arm glove for puerperal intrauterine exploration. AB - OBJECTIVE: To design a long arm glove that can be used within a puerperal uterus to prevent the health-care worker contracting HIV from an infected patient. The designed long arm glove should be cheap (affordable) and readily available in low resource centres and must have proven sterility assurance and tensile strength to confer protection for the health worker. DESIGN: Prospective study. SETTING: Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria from 1st December 2006 to 31st May, 2007. SUBJECTS: Fifty medical students of both sexes were selected randomly and the average length from the styloid process to the mid upper-arm of each was measured and the average length was later determined. This was to determine the length of the low density polyethylene long arm gloves to be made from virgin polyethylene material. Consecutive cases of patients with retained placentae in the puerperium who were scheduled for manual removal of the placenta within the period. RESULTS: Packs of low density polyethylene (LDPE) long arm gloves were made from virgin polyethylene material. When subjected to bacteriological analysis, three out of four glove packs were contaminated with Staphylococcus, Bacillus and Klebsiella species of bacteria. Gamma irradiation ranging from 28.133 to 83.35 kiloGray of gamma-irradiation (kGy) sterilised all the gloves as postirradiation glove specimens showed no bacterial contamination. However, at doses up to 50 kGy gamma irradiation caused "strengthening" of the polyethylene gloves. While at doses greater than 50 kGy, gamma irradiation caused "embrittlement" of the material. Thus, 50 kGy of gamma irradiation was found to be an ideal dose to strengthen and to sterilise the glove for usage. The sterilized gloves were found to be effective when used in consecutive cases of retained placenta in protecting the health care workers (HCW) from contamination by possibly HIV contaminated blood. CONCLUSION: There is a risk of contracting HIV for the health-care worker while carrying out a procedure within the puerperal uterus. The low density polyethylene arm glove was designed to prevent this in low resource centres as it would be affordable, available, with proven sterility assurance and mechanical properties to confer protection for the health care worker. PMID- 19133426 TI - Laparoscopic myomectomy of a large pedunculated fibroid: case report. AB - A 37-year-old para 0 + 0 presented with abdominal pain and progressive abdominal swelling. She had no menstrual complains. Abdominal examination revealed a large 30 week non-tender, firm and mobile mass. A pelvic/abdominal ultrasound confirmed a 15.7 x 9.9 cm pedunculated fibroid arising from the uterine fundus. A laparoscopic myomectomy was successfully undertaken, and the morcellated fragments weighed 1490 grams. PMID- 19133428 TI - [Operative technique and preliminary results of arthroscopic reconstruction of posterior cruciate ligament]. AB - Injuries of knee generally result in damage to major ligaments which stabilize this joint. Clendenin observed that 20% failure of ligaments of knee it is PCL rapture. A consequence of this type injuries is mechanical and functional instability of the knee joint. The goal of this article is to present our technique of arthroscopic reconstruction of PCL. We also present preliminary results of our treatment options we have applied in 14 surgery patients. The study material consisted of 14 patients who underwent surgical treatment for PCL injuries in the Department of Pediatric Orthopedics and Traumatology in Poznan. The arthroscopic reconstructions of PCL in all 14 patients have been performed by first author with the same operative technique. Knee laxity we assessed before and after PCL reconstructive surgery using arthrometer "Rolimeter" (Aircast). Knee ligament rating scales (Lyscholm and IKDC 2000) have been used to perform subjective, functional assessment of knee joint. The follow up was about 2 years. We fund statistical important improvement in all tested data (p = 0.001). The arthroscopic reconstruction of PCL with autogenic hamstring graft improves function and stability of knee joint. Endurance and size of hamstring grafts are sufficient to restore good mechanical and functional stability of knee joint. PMID- 19133427 TI - [Knee dislocation: concurrent arthroscopic ACL and PCL reconstruction - operating technique]. AB - Injuries involving knee luxation or subluxation generally result in damage to two or three out of the four major ligaments which stabilize this joint. Past treatment of this type of multiligament injuries, consisting in 6- to 12-week immobilization in a plaster cast, does not allow for restoration of the normal, anatomical course of these ligaments. A consequence of this type of healing, which involves the formation of random scars in place of ligaments, is mechanical and functional instability of the knee joint. The goal of this article is to present a classification of multiligament knee injuries and surgical treatment options. We also present our own technique of arthroscopic, concurrent reconstruction of knee joint ligaments. The study material consisted of 20 patients who underwent surgical treatment for multiligament knee injuries in the Department of Pediatric Orthopedics and Traumatology in Poznan. We performed concurrent arthroscopic reconstruction of ACL and PCL in all 20 patients. In addition, reconstruction of the medial complex was performed in 7 patients, and lateral complex was reconstructed in another 7 patients. In three cases we stitched the medial meniscus using a Smith & Nephew FasT-Fix system, and partial menisectomy was performed in 2 patients. The details of the proposed concurrent arthroscopic ACL and PCL reconstruction are extensively discussed. One of the advantages of this operating technique is the exclusive use of autogenous ST and GR tendon grafts for reconstruction of all damaged knee joint structures. The presented operating technique makes possible the reconstruction of all damaged knee joint ligaments in less than two hours, which in turn allows the surgery to be performed in an ischemic setting using an Esmarch band around the limb. PMID- 19133429 TI - [Inability to work before and after operation for carpal tunnel syndrome]. AB - INTRODUCTION: Carpal tunnel syndrome is the most common compressive neuropathy, affecting approximately 5.8% women and 0.6% men. In consideration of its commonness and occurrence in persons in employment age it is considered a cause of significant work absence. Operative treatment of the syndrome results in temporary decreasing of the power and dexterity of the hand, what is also a cause of inability to work. There is lack of information in Polish literature about the dimension of this problem. The objective of this study was to determinate a work absence caused by carpal tunnel syndrome, both before and after the operation. PATIENTS AND METHODS: Fifty-six employed patients, 44 women and 12 men in a mean age of 49 years (range 27-63) who underwent a mini-invasive carpal tunnel release were analyzed. Patients were asked for duration of the disease and time off work before the operation and were followed-up for 6 months with a time off work to be noted. At 6 months, a self-assessment of the satisfaction with the result was performed in a simple four-grade scale. RESULTS: A mean duration of the disease in 56 employed patients was 34 months (range 2 months-20 years). Prior to the surgery, 22 patients (39%) were on sickness related sick leave for an average 2.9 months (range 3-12), After the operation all patients availed of sick leave for a mean of 2.3 months (range 1-6). Eight (14%) of the employed patients did not return to work in a 6 months follow-up for various reasons, but mostly due to weakness and reduced dexterity of the involved hand. None of the patients complained of symptoms similar to the pre-operative and in none any complication occurred. In subjective evaluation of the effectiveness of the surgery, comparing to the status before operation 31 patients (26%) were completely free of pain and other symptoms, in 80 (66%) complaints significantly reduced, in 5 (4%) remained the same and in 5 (4%) deteriorated. Statistical analysis revealed that 22 patients who availed of sick leave prior to surgery, were statistically significantly longer off work (a mean of 3 months) than 34 patients who did not were on sick leave (a mean of 1.8 months). Similarly, in 8 patients who did not return to work, 7 availed of sick leave before the operation and only one did not. CONCLUSION: Our results show that carpal tunnel syndrome is in Poland a cause of considerable work absence. In comparison to other countries, time off work after surgery is relatively long and a proportion of patients did not return to work within 6 months, regardless that no complications occurred in these patients. We intuitively think, that a main cause of a long lasting time work off in majority of patients after carpal tunnel release is an unwillingness but no an inability to work. PMID- 19133430 TI - [Treatment of the radial shaft loss with pedicled graft from the ulna--a case report]. AB - A case of 68-year-old woman with severe deformation of the left forearm and disability of the hand as a consequence of total dissociation and partial loss of radial shaft, after fracture sustained one year earlier is reported. The restoration of the length and shape of the radius was performed with the use of 4 cm long fragment of the ulna pedicled on the interosseous membrane, fixated with plate. Normal shape of the radius was achieved and the hand was held in the axis of the forearm. Bone healing was achieved at one year, and at the 2.5 years follow-up the patient was free of pain, had full fingers movement, full rotation of the forearm, total grip strength of 16 kG (64% of the power of the healthy hand) and subjective evaluation of the dexterity with DASH scale was 25 points, suggesting a mild loss of function. PMID- 19133431 TI - [Forearm tumor of the soft tissue inflammatory-like etiology]. AB - We write about case of 60-year-old man with a forearm tumor about 7 cm large. The change revealed to have inflammatory-like etiology. The surgery made a good result for the health and quality of life for the patient. The histopatlogy excluded the neoplastic etiology. We observed phenomenas big tumor and no pathological indication in function of medial and ulnar nerve. PMID- 19133432 TI - [Latissimus dorsi transfer for treatment of irreparable rotator cuff lesions]. AB - The paper presents results of treatment of irreparable rotator cuff lesions by means of latissimus dorsi transfer. Material consists of 10 patients treated in our center in 2003-2006. 9 male and 1 female patients were operated. The av. age was 56.3 (27-65). Follow-up period was 15.3 months on average. Evaluation included following measurements: range of motion, pain, UCLA score. 4 patients were evaluated also by ultrasound imaging and EMG. Average increase of active flexion was 70 degrees, active abducion--71 degrees, active external rotation--23 degrees. Pain was significantly diminshed in all patients. The result of treatment in our study group are similar to that presented in literature. Based on this paper latissimus dorsi transfer can be recommended as a technique for traetment of irreparable rotator cuff PMID- 19133433 TI - [Adam Gruca (1893-1983)]. AB - Adam Gruca is one of most eminent and most well-known Polish orthopaedists. His contribution to the development of this branch of medicine is huge, and innovations, of which was an author remain current to this day. In the article author introduced both his unusual, often the dramatic life, as well as the wonderful academic career. PMID- 19133434 TI - [Famous figures of the Poznan orthopaedics of the period of the occupation and post-war years. Coryphees of Polish orthopaedics]. AB - In this article author presents, from a perspective of own memories is portraying persons which he met in his professional activity. They participated in forming the orthopaedics in Poznan and different nooks of Poland. He resembles their, often very dramatic, fates and the influence they had on Polish medicine reviving after the II world war. With the special attention he is reminding one of most well-known and valued celebrities of the Polish orthopaedics professor Wiktor Dega. PMID- 19133435 TI - Acute traumatic hemarthrosis of the adult's knee--a diagnostic options in arthroscopic era. Literature review. AB - The review presenting the most common causes of acute knee hemarthrosis in adults, discuss diagnostic accuracy, advantages and disadvantages as well as cost effectiveness assessment of physical examination, magnetic resonance imaging and artrhroscopy as diagnostic tools in appraisal of acute injured knee. PMID- 19133436 TI - [Bluetongue virus serotype 8 in healthy young calves]. AB - Following the detection in Northern Ireland of bluetongue serotype-8 (BTV-8) PCR positive calves born from PCR-negative but seropositive heifers imported from the Netherlands, an investigation was started to determine whether PCR-positive calves were born in the first quarter of 2008 in the Netherlands. Blood samples were collected on 43 dairy farms from 388 cow-calf combinations. All calves were at least 10-days-old and had been born after 1 January 2008. In total, 229 cows had had a BTV-8 infection, and from these cows 37 PCR-positive calves were born (16.2%). If these calves are viraemic, they can contribute to BTV-8 overwintering in North-West Europe. PMID- 19133437 TI - [Veterinary dentistry: an update 2008]. AB - Rooted in human dentistry, veterinary dentistry has developed steadily in the Netherlands since the 1980s and is now recognized as an essential discipline of veterinary medicine. The availability of specialized tools and techniques has led to improved treatment outcomes and results, with the choice of treatment being largely determined by the functionality of the dentition and the costs involved. Domestic animals and horses with dental problems should be referred to dental veterinarians. The Working Group Veterinary Dentistry in the Netherlands is an association for skilled veterinarians with professional dental equipment at their disposal. PMID- 19133438 TI - [Punctured through]. PMID- 19133439 TI - [Vaccination against bluetongue serotype 8 in the Netherlands]. PMID- 19133440 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, etc. Veterinary cream pot from the government veterinary school]. PMID- 19133441 TI - [MRSA]. PMID- 19133442 TI - [MRSI and the veterinary clinic]. PMID- 19133443 TI - [Supervisory veterinarian with the food safety authorities: "supervising animal welfare is becoming more and more important"]. PMID- 19133444 TI - [Cash is king]. PMID- 19133445 TI - ["Meeting of real patients is very motivating"]. PMID- 19133446 TI - [Veterinary ergonomics: a matter of life style]. PMID- 19133447 TI - [New committee DSK (Veterinary Student Circle)]. PMID- 19133448 TI - A binomial and species-independent approach to trap capture analysis of flying insects. AB - Traps for monitoring of flying insect pests constitute a critical part of integrated pest management strategies. However, interpretation of trap captures is hampered by 1) factors associated with the performance of traps (i.e., lure, trap design, placement); 2) an often poorly defined relationship between trap captures and population density; and 3) interpretation approaches being highly specific to a certain insect species, trapping method, or trapping environment. The main purpose of this study was to identify a trap capture interpretation approach with little sensitivity to characteristics specific to a given data set, which would allow easier comparison of trapping data sets and make it easier to standardize sampling plans across insect pests and trapping environments. Based on fits of trapping data sets to standard distributions (normal, Poisson, and negative binomial), evaluations of the index of aggregation, k, and linear regression coefficients from Taylor's power law, we concluded that these characteristics varied considerably among data sets, which means that enumerative sampling plans may not be appropriate. Across 13 trapping data sets of six insect species, we showed a consistent nonlinear relationship between average trap captures and number of traps with zero captures and that the k can be stabilized by converting trapping data into binomial data. A trap interpretation approach based on number of zero captures is both easy to use, was found to be species independent, and means that it may be possible to establish meaningful and reliable action thresholds based on trap captures of flying insects. Although developed using trapping data from food facilities, this approach may have application to trapping data from other environments as well. PMID- 19133449 TI - Genetic profiling to determine potential origins of boll weevils (Coleoptera: Curculionidae) captured in a Texas eradication zone: endemicity, immigration, or sabotage? AB - Thirty-seven boll weevils, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), were captured in pheromone traps near Lubbock, TX, in the Southern High Plains/Caprock eradication zone during August-October 2006. No boll weevils had been captured in this zone or neighboring zones to the north earlier in the year, and only very low numbers had been captured in neighboring zones to the south and east. Therefore, the captures near Lubbock were unexpected. Five of the weevils captured the last week of August were preserved and genotyped at 10 microsatellite loci for comparison with a database of genotypes for 22 boll weevil populations sampled from eight U.S. states and four locations in Mexico. The Lubbock population itself is an unlikely source, suggesting that the captured weevils probably did not originate from a low-level endemic population. Populations from eastern states, Mexico, and Big Spring, TX, can be confidently excluded as potential source regions. Although the Weslaco and Kingsville, TX, areas cannot be statistically excluded, they are unlikely sources. The most likely sources are nearby areas in New Mexico, TX, or southwest Oklahoma, or from areas of eastern Texas represented by Waxahachie and El Campo populations. Together, genetic and circumstantial evidence suggest either that the trapped boll weevils are the offspring of alone mated female that immigrated from eastern Texas earlier in the summer or that weevils originally captured near Waxahachie but now long-dead were planted in the traps by a disgruntled employee of the eradication program. PMID- 19133450 TI - Validation of a method using queenless Bombus terrestris micro-colonies for testing the nutritive value of commercial pollen mixes by comparison with queenright colonies. AB - The nutritive value of pollen blends purchased by bumble bee producers from beekeepers is a key factor of successful mass rearing. We have already devised a method for quality ranking of pollen diets. It was a 1-mo bioassay using queenless micro-colonies of three callow workers (Bombus terrestris L.; Apidae: Bombinae). In the current study, we tested three pollen diets through the micro colony method and compared the results to the development of queenright colonies supplied with the same diets. The three diets were "A," a spring blend with a protein rate of 15.7% and a dominance of Corylus avellana L.(46%) and Buxus sempervivens L. (35%); "B," a summer assemblage with alower crude protein content (14.4%), with a dominance of Helianthus annuus L. (37%), Zea mays L. (35%), and Poaceae (21%); and "C," a 50/50 mixture by weight of A and B, with a crude protein content of 15.8%. In micro-colonies, the more sensitive parameter for diet ranking was the mean weight of a larva, whereas in queenright colonies, the discrimination between treatments was enabled by considering the body size of new queens (weight and length of the radial cell) and the slope of the sigmoidal curve of the pollen consumption at the inflection point reflecting brood growth. Both testing methods assigned the lower rank to diet B compared with diet A. Despite the closeness of the three diets in terms of nitrogen content, we concluded that micro-colonies were a good estimate of colony development when nutritive value of pollen was tested. PMID- 19133451 TI - Abnormal foraging behavior induced by sublethal dosage of imidacloprid in the honey bee (Hymenoptera: Apidae). AB - Although sublethal dosages of insecticide to nontarget insects have never been an important issue, they are attracting more and more attention lately. It has been demonstrated that low dosages of the neonicotinoid insecticide imidacloprid may affect honey bee, Apis mellifera L., behavior. In this article, the foraging behavior of the honey bee workers was investigated to show the effects of imidacloprid. By measuring the time interval between two visits at the same feeding site, we found that the normal foraging interval of honey bee workers was within 300 s. However, these honey bee workers delayed their return visit for > 300 s when they were treated orally with sugar water containing imidacloprid. This time delay in their return visit is concentration-dependent, and the lowest effective concentration was found to be 50 microg/liter. When bees were treated with an imidacloprid concentration higher than 1,200 microg/liter, they showed abnormalities in revisiting the feeding site. Some of them went missing, and some were present again at the feeding site the next day. Returning bees also showed delay in their return trips. Our results demonstrated that sublethal dosages of imidacloprid were able to affect foraging behavior of honey bees. PMID- 19133452 TI - Brood pheromone effects on colony protein supplement consumption and growth in the honey bee (Hymenoptera: Apidae) in a subtropical winter climate. AB - Fatty acid esters extractable from the surface of honey bee, Apis mellifera L. (Hymenoptera: Apidae), larvae, called brood pheromone, significantly increase rate of colony growth in the spring and summer when flowering plant pollen is available in the foraging environment. Increased colony growth rate occurs as a consequence of increased pollen intake through mechanisms such as increasing number of pollen foragers and pollen load weights returned. Here, we tested the hypothesis that addition of brood pheromone during the winter pollen dearth period of a humid subtropical climate increases rate of colony growth in colonies provisioned with a protein supplement. Experiments were conducted in late winter (9 February-9 March 2004) and mid-winter (19 January-8 February 2005). In both years, increased brood area, number of bees, and amount of protein supplement consumption were significantly greater in colonies receiving daily treatments of brood pheromone versus control colonies. Amount of extractable protein from hypopharyngeal glands measured in 2005 was significantly greater in bees from pheromone-treated colonies. These results suggest that brood pheromone may be used as a tool to stimulate colony growth in the southern subtropical areas of the United States where the package bee industry is centered and a large proportion of migratory colonies are overwintered. PMID- 19133453 TI - Inheritance of resistance to Acarapis woodi (Acari: Tarsonemidae) in crosses between selected resistant Russian and selected susceptible u.s. honey bees (Hymenoptera: Apidae). AB - The pattern of inheritance of tracheal mite resistance in selected Russian bees was determined in bioassays and in samples from field colonies. Resistant colonies of Russian origin and colonies selected for high susceptibility in the United States were used to generate divergent parental populations. Seven groups of F1 colonies were produced by crossing queens and drones from these selected resistant Russian and selected susceptible populations. In a series of bioassays with young workers exposed in infested colonies, average mite abundance (female mites per worker) in F1 colonies was intermediate (1.04 +/- 0.13 [mean +/- SE]) and significantly different from that of both resistant Russian (0.74 +/- 0.13) and selected susceptible (1.57 +/- 0.13) colonies. Colonies representing the three populations were established in two apiaries in July 2005. Colonies surviving with original queens after 10 mo had mite prevalences supporting the findings of the bioassay. All three resistant colonies had undetectable mite levels, whereas prevalences in four F1 colonies ranged from 0 to 53%, and in 10 susceptible colonies ranged from 0 to 90%. Tracheal mite resistance in Russian bees is likely polygenic, but there may be a number of genes with major dominance interacting with minor genes. Use of selected Russian queens mated with Russian drones or with drones from unknown sources is beneficial for beekeeping in areas with persistent problems with tracheal mite infestation. PMID- 19133454 TI - Cold storage of adult Gonatocerus ashmeadi (Hymenoptera: Mymaridae) and effects on maternal and progeny fitness. AB - Storage of Conatocerus ashmeadi Girault (Hymenoptera: Mymaridae) adults at 2, 5, and 10 degrees C showed that these parasitoids do not survive at 2 degrees C for 5 d, and exposure to 5 and 10 degrees C shortens their life span. The lethal time (LT) 50 (i.e., length of storage time for 50% wasp survival) at 5 degrees C was 14 d for males and approximately 29 d for females, whereas at 10 degrees C was 32 and 39 d, respectively. Effects of adult storage at 10 degrees C on other factors indicating fitness, such as fecundity, developmental time, parasitism, emergence, and sex ratio, were examined on female wasps and their progeny at 10-d intervals for up to 60 d. Glassy-winged sharpshooter, Homalodisca vitripennis (Germar), eggs were used as hosts for propagation of this wasp and for assessing its oviposition and fecundity. Increasing adult storage time decreased the length of the ovipositional period for the maternal generation, and oviposition was decreased by 90% after 60-d storage. A significant reduction in maternal lifetime fecundity occurred after 20-d storage and in the incidence of parasitism after 40 d. We also found a carryover effect caused by storage of the maternal generation that was expressed in the F1 generation. When cold storage of the adult parents was > or = 20 d, we observed delayed development, decreased fecundity, reduced longevity, and increased male production occurring in the F1 generation. Reduced fitness of the F1 generation was also expressed as a decrease in net reproductive rate (R(o)) and an increase in mean generation time (T(c)). However, none of these deleterious effects were evident in the F2 progeny that descended from grandparents that had experienced cold storage. Damage caused by indirect chilling injury and/or induced maternal aging occurring during storage can account for the decreased fitness of maternal and F1 generations. Providing that the limits of cold tolerance of G. ashmeadi as defined in this study are not exceeded, our results show that short-term cold storage of adults could be used in a mass-rearing program. PMID- 19133455 TI - Development of a strategy for selective collection of a parasitoid attacking one member of a large herbivore guild. AB - Selectively collecting a single natural enemy species that parasitizes one member of a guild of herbivores that attack the same host plants can be a challenging problem during development of biological control programs. We present here a successful strategy for the collection of a strain of the egg parasitoid Avetianella longoi Siscaro (Hymenoptera: Encyrtidae), that parasitizes eggs of the longhorned borer Phoracantha recurva Newman (Coleoptera: Cerambycidae). This cerambycid is one member of a large guild of woodborers that simultaneously infest dying and fallen Eucalyptus in Australia, and it has become a major pest of Eucalyptus in many areas of the world where Eucalyptus has been introduced. Adult P. recurva of both sexes were caged on freshly cut Eucalyptus logs, and the resulting egg masses were marked and then left exposed to natural parasitization in the field. Parasitized egg masses were then harvested and held in the laboratory until adult parasitoids emerged. Parasitoids were identified as A. longoi by morphological comparisons with reference specimens, and with molecular markers. This strain of A. longoi readily accepted and had high survival rates in eggs of P. recurva. In contrast, the strain of A. longoi that has been used for biological control of P. semipunctata in California since the 1990s strongly prefers eggs of eucalyptus longhorned borer, Phoracantha semipunctata (F.) (Coleoptera: Cerambycidae), and has relatively low rates of survival to adulthood in eggs of P. recurva. The causes of these behavioral and physiological differences between the two strains are not yet known. PMID- 19133456 TI - Effects of production practices on the abundance of quarantine pests in Dracaena marginata in Costa Rican production fields. AB - To mitigate risks of invasive pests, international nursery exporters may soon be expected to participate in clean stock programs that reduce the probability of shipping crops that contain pests of quarantine significance. We worked with Costa Rican producers of Dracaena marginata Lam. (Ruscaceae) to identify critical pests, as well as the field conditions and production practices that could be explored for lowering pest abundance in a clean stock program. We sampled the abundance of leafhopper and katydid eggs, armored scales, and snails on leaves using a 10- by 10-m grid in a total of 34 production plots located in two distinct growing regions during the rainy and dry seasons of 2006. Growers were surveyed to determine the production practices in each field. Populations of leafhoppers and snails increased during the rainy season, whereas armored scale populations reached higher densities in the dry season. Fertilizer applications were associated with a greater abundance of leafhoppers, the keystone pests in the system. In contrast, abundances of armored scales and katydids were somewhat lower in fertilized fields. Insecticide applications targeting leafhoppers succeeded in reducing their populations, but secondary outbreaks of armored scales and katydids were observed. Increased frequency of herbicide application was associated with lower populations of leafhoppers with no significant effects on the other herbivores. Although effects of specific production practices in a given plot are confounded, the effects of fertilizer, insecticide, and herbicide use on leafhoppers are consistent with those predicted by literature on this pest. For this reason, the 10- by 10-m sampling grid is sensitive enough for use in controlled studies that evaluate effects of specific practices on pest abundance. As such, it is likely to be a useful tool for developing a clean stock program for D. marginata in Costa Rica. PMID- 19133457 TI - Low temperature phosphine fumigation for postharvest control of western flower thrips (Thysanoptera: Thripidae) on lettuce, broccoli, asparagus, and strawberry. AB - U.S. exported lettuce, broccoli, asparagus, and strawberries often harbor western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), a quarantined pest in Taiwan, and therefore require quarantine treatment. Fumigation with diluted pure phosphine at a low temperature of 2 degrees C was studied to control western flower thrips and to determine effects on the quality of the treated products. Total thrips control was achieved in > or = 18-h fumigation treatments with > or = 250 ppm phosphine. One day fumigation treatment with 1,000 ppm phosphine was tested on lettuce and broccoli. One-day fumigation treatments with 500 ppm and 1,000 ppm phosphine were tested on asparagus and strawberry. Visual quality of lettuce, broccoli, and asparagus was evaluated after 2-wk posttreatment storage. Strawberry quality was evaluated immediately after fumigation and after 1-wk posttreatment storage. For all the products, there were no significant differences between the treatments and the controls in postharvest quality, and there were no injuries caused by the fumigation treatments. Therefore, phosphine fumigation at low temperature was promising for postharvest control of western flower thrips on lettuce, broccoli, asparagus, and strawberry. PMID- 19133458 TI - Effect of wild flowers on oviposition of Hippodamia variegata (Coleoptera: Coccinellidae) in the laboratory. AB - Marginal vegetation in crops is very important for natural enemies and their pest control capacity. The effects of Brassica nigra L. (Brassicaceae), Daucus carota L. (Apiaceae), and Sonchus oleraceous L. (Asteraceae) flowers as supplemental food on the number of eggs laid during 7 d and on the preoviposition time in Hippodamia variegata (Goeze, 1777) were studied in the laboratory under conditions of several densities of Acyrthosiphon pisum (Harris, 1776). The results show the presence of flowers of Brassica and Sonchus increased egg production 1.44X and doubled the pre-oviposition period (2.13X). This suggests that the availability of flowers of Brassica and Sonchus as supplemental foods (pollen and nectar) in the marginal vegetation of crops can serve to improve reproductive performance of H. variegata, specifically under conditions of prey limitation. Thus, the increase in fitness of this predator allows a better response to changes in pest density. PMID- 19133459 TI - Effect of temperature on the life history of the mealybug Paracoccus marginatus (Hemiptera: Pseudococcidae). AB - Effect of temperature on the life history of the mealybug Paracoccus marginatus Williams & Granara de Willink (Hemiptera: Pseudococcidae) was investigated in the laboratory. P. marginatus was able to develop and complete its life cycle at 18, 20, 25, and 30 +/- 1 degrees C. At 15, 34, and 35 degrees C, the eggs hatched after 27.5, 5.9, and 5.5 d of incubation, respectively, but further development of the first-instar nymphs was arrested. No eggs hatched at 37 degrees C. The developmental time for egg to adult was the longest at 18 degrees C for both males and females. Approximately 80-90% of the eggs survived between 20 and 30 degrees C. The highest fecundity was at 25 degrees C with each female producing an average of 300 eggs. Adult longevity, and preoviposition and oviposition periods increased with decreasing temperature up to 25 degrees C. The proportion of females was approximately 42% at 25 degrees C and was between 70 and 80% at 18, 20, and 30 degrees C. Adult males and females required 303.0 and 294.1 degree days (DD), respectively, to complete their development. The estimated minimum temperature thresholds for the adult males and females were 14.5 and 13.9 degrees C, respectively. For adult males, the estimated optimum and maximum temperature thresholds were 28.7 and 31.9 degrees C; and for adult females, they were 28.4 and 32.1 degrees C, respectively. The ability of P. marginatus to develop, survive, and reproduce successfully between 18 and 30 degrees C suggests that it has the capability to develop and establish in areas within this temperature range. PMID- 19133460 TI - Lethal and sublethal effects of endosulfan on Apolygus lucorum (Hemiptera: Miridae). AB - Various pesticides directly impact insect pest populations through acute toxicity (lethal) effects, but they also cause a range of indirect (sublethal) effects on pest physiology or behavior. In this study, we describe both lethal and sublethal effects of endosulfan on the survival, development, and fecundity of Apolygus lucorum Meyer-Dur (Hemiptera: Miridae), an important pest of Bacillus thuringiensis (Bt) cotton, Gossypium hirsutum L., in China. We quantify pesticide effects on adult mirids as well as on subsequent (F1, F2) generations. Toxicity bioassays determined the median lethal concentration (LC50) for adult A. lucorum as 0.58 ppm. To evaluate the sublethal effect of endosulfan, we used LC20 and LC40 concentrations at 0.30 ppm and 0.48 ppm, respectively. Sublethal concentrations of endosulfan significantly reduced longevity and fecundity of female adults, but slightly affected male longevity. We also found sublethal effects on the F1 generation, through a reduction of egg hatching rate, nymphal mortality, adult molting rate, and adult fecundity, and on the F2 generation, through a decrease of egg hatching rate. These results show that endosulfan has far greater effects on A. lucorum than estimated by acute toxicity tests, and the effects of this pesticide on surviving mirids can be comparatively long-lived. This pesticide likely causes severe disruptions of A. lucorum population dynamics, with single endosulfan application, invoking a great reduction of population growth of resident mirid populations. We discuss implications of this work for control programs for A. lucorum on Bt cotton. PMID- 19133461 TI - Impact of decreasing ratios of insecticide-treated seed on flea beetle (Coleoptera: Chrysomelidae, Phyllotreta spp.) feeding levels and canola seed yields. AB - Field studies were conducted at two locations on the Canadian prairies to investigate use of reduced ratios of insecticide-treated seed in controlling flea beetle (Coleoptera: Chrysomelidae, Phyllotreta spp.) damage to canola (Brassica napus L. and Brassica rapa L.). Five treatments were evaluated: bare seed control, fungicide-only (0X), and three ratios of insecticide plus fungicide in proportions of all (1X), two thirds (0.67X), or one third (0.33X) of the seeds coated with insecticide. Decreasing treated seed ratios by one third had no consistent deleterious effects on flea beetle damage, seedling growth, plant density, seed yield, or net cash return. Flea beetle injury to seedlings in the 1X treatment was similar to that of seedlings in the 0.67X treatment, with only two exceptions, and it was almost always lower than that of seedlings without insecticide. The 0.33X treatment generally had flea beetle feeding levels between those of the two high and the two noninsecticide treatments. Plant stand and seedling growth rates with 1X and 0.67X treatments were similar and higher than with bare seed or fungicide-alone treatments. Seed yields were inversely proportional to flea beetle feeding levels. Under very heavy flea beetle feeding, seed yields and net cash returns were highest in 1X plots, but when flea beetle feeding pressure was less extreme and canola growing conditions were favorable, 0.67X seed yields and profits from them were comparable to those in 1X treatments. On an economic basis, currently there is no advantage to decreasing the level of insecticide treated canola seed, but other considerations may affect this assessment. PMID- 19133462 TI - Field response of Dendroctonus frontalis (Coleoptera: Scolytinae) to synthetic semiochemicals in Chiapas, Mexico. AB - Dendroctonus frontalis Zimmermann (Coleoptera: Scolytinae) is the most serious pest of pines (Pinus spp.) in Mexico. Conspecifics are attracted to trees undergoing colonization by the aggregation pheromone frontalin, which is synergized by odors of pine oleoresin released from beetle-damaged host tissue. Synthetic racemic frontalin combined with turpentine has been the operational bait used in traps for monitoring populations of D. frontalis in Mexico as well as the United States. Recently, racemic endo-brevicomin has been reported to be a synergist of the frontalin/turpentine bait and as an important component of the aggregation pheromone for D. frontalis populations in the United States. To determine whether racemic endo-brevicomin also might function as an aggregation synergist for the geographically isolated D. frontalis populations of Central America and Mexico, we performed a field trapping trial in Lagunas de Montebello National Park, Chiapas, Mexico, during July and August 2007. The combination of endo-brevicomin (placed either directly on the trap or 4 m away) plus racemic frontalin and turpentine caught at least 5 times more D. frontalis of both sexes than did turpentine either alone or in combination with either frontalin or endo brevicomin. The addition of endo-brevicomin to the frontalin/turpentine bait also increased the proportion of females trapped. We conclude that the addition of endo-brevicomin might substantially improve the efficiency of the frontalin/turpentine bait for monitoring of D. frontalis in Central America and Mexico. We discuss factors that reconcile our results with previous studies that reported endo-brevicomin to be an attractant antagonist for populations of D. frontalis in Mexico and Honduras. PMID- 19133463 TI - High release rate 3-methylcyclohex-2-en-1-one dispensers prevent Douglas-fir beetle (Coleoptera: Curculionidae) infestation of live Douglas-fir. AB - The Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (Coleoptera: Curculionidae), antiaggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH), has been used by natural resource managers and landowners to protect high-value, high risk trees from Douglas-fir beetle infestation throughout the western United States since 2000. Labor is a major portion of the cost of MCH treatments. MCH is applied by walking through treatment areas and stapling the formulated pheromone in bubble capsules to trees and other objects on a regular grid pattern. Reducing the number of MCH release points and increasing the distance between them could significantly reduce labor costs, particularly in areas with steep terrain or large volumes of woody debris that could impede the movement of applicators. This study compared the standard MCH application method with a method releasing MCH at a 3 times higher rate and placed at three times fewer release points per unit area. Treatments were applied to 2-ha plots simulating an operational application. Aggregation pheromone-baited traps were placed at plot centers to ensure that dispersing adult beetles would be present on all plots. Both MCH treatments were equally effective at preventing the infestation of live Douglas fir, Pseudotsugae menziesii (Mirbel) Franco, trees (> or = 30 cm diameter at breast height). These results confirm that MCH formulated to release at three times the current standard rate and placed at 3 times fewer points per unit area can effectively prevent the infestation of live Douglas-fir. The new treatment will significantly reduce the labor cost of MCH applications making them feasible for areas that may have previously been marginal economically. PMID- 19133464 TI - Influence of trap placement and design on capture of the emerald ash borer (Coleoptera: Buprestidae). AB - The key to an effective pest management program for the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera Buprestidae), is a survey program equipped with tools for detecting and delimiting populations. We studied the effects of trap design, color, and placement on the efficacy of sticky traps for capturing the emerald ash borer. There were significant differences in trap catch along a transect gradient from wooded to open field conditions, with most beetles being caught along the edge, or in open fields, 15-25 m outside an ash (Fraxinus spp. L.) (Oleaceae) woodlot. Greater emerald ash borer catch occurred on purple traps than on red or white traps. Traps placed in the mid-canopy of ash trees (13 m) caught significantly more beetles than those placed at ground level. We also describe a new trap design, a three-sided prism trap, which is relatively easy to assemble and deploy. PMID- 19133465 TI - Abiotic factors and trap design modulate the performance of traps used to monitor the plum curculio. AB - All published studies on effects of abiotic factors on plum curculio, Conotrachelus nenuphar (Hersbt), adults have taken a retrospective approach. Here, we present the results of experiments where factors and their levels were determined and controlled a priori. We compared the effectiveness of miniature pyramidal traps (45 by 20 by 20 cm) constructed of four kind of materials--wood, geotextile, nylon screening, and corrugated plastic--to monitor overwintered and summer adults of univoltine plum curculio. We also studied the effects of photoperiod, temperature, wind, and rain on pyramidal trap effectiveness. The experiments, which were replicated over time, were done in two controlled chambers that were divided into four sections, corresponding to simulated combinations (wind or no wind/rain or no rain). The temperatures tested (15, 20, and 25 degrees C) were randomly assigned in the chambers. During scotophase, geotextile traps captured significantly more overwintered and summer adults than traps made of other materials. The maximum proportion of captures (for overwintered and summer adults) during photophase was obtained at 25 degrees C, and it was significantly different than captures at 15 and 20 degrees C. During scotophase, significantly more overwintered and summer plum curculios were caught at 20 and 25 degrees C than at 15 degrees C. Our experiments demonstrated that geotextile is a good alternative to wooden pyramidal trap. Our results suggest that captures were higher 1) at night, 2) during warmer periods (20 and 25 degrees C), 3) when wind velocity was low and 4) during or shortly after rainfall, and 5) that photoperiod is a factor having an important predictive value for plum curculio captures. PMID- 19133466 TI - Seasonal phenology and management of Tomarus subtropicus (Coleoptera: Scarabaeidae) in St. Augustinegrass. AB - The grub Tomarus subtropicus Blatchley (Coleoptera: Scarabaeidae) is one of the most destructive pests of turfgrass and sugarcane (Saccharum spp.) in southern Florida, but its life cycle has only been described in sugarcane production. Preventive insecticides applied against T. subtropicus in turfgrass when adult activity and oviposition were expected in sugarcane have necessitated subsequent curative treatments. Thus, the seasonal activity of T. subtropicus was monitored using UV blacklight traps and by sampling soil in infested St. Augustinegrass lawns in Cape Coral and Punta Gorda, FL, during 2005-2006. Several preventive and curative insecticides and entomopathogenic nematode species were evaluated in laboratory, greenhouse and field tests. T. subtropicus is univoltine in both sugarcane and St. Augustinegrass, but adult flight peaked approximately 1 mo later in St. Augustinegrass than in sugarcane. Halofenozide, clothianidin, and imidacloprid were effective preventive control products, and carbaryl and trichlorfon were satisfactory curative insecticides. The nematodes Steinernema scarabei (Stock & Koppenhofer), S. glaseri (Steiner), and Heterorhabditis bacteriophora (Poinar) were most effective against T. subtropicus grubs, and second instars were more susceptible compared with third instars. Improved knowledge of the biology and effectiveness of different management options for T. subtropicus will help minimize excessive pesticide use, especially in coastal areas of Florida. PMID- 19133467 TI - Developing wax-based granule formulations for mating disruption of oriental beetles (Coleoptera: Scarabaeidae) in turfgrass. AB - Oriental beetle, Anomala orientalis Waterhouse (Coleoptera: Scarabaeidae), is a pest of turfgrass that may be controlled by applications of synthetic pheromone (Z)-/ (E)-7-tetradecen-2-one to disrupt mating. Laboratory experiments were conducted to determine release profiles of pheromone from experimental wax-based granules, a proprietary wax granule, and rubber septa commonly used in pheromone traps. Rubber septa loaded with 10, 100, and 300 microg per septum provided steady rates of pheromone release (zero-order) over 4 wk of laboratory evaluation (total = 1.1, 9.0, and 26.9 microg/4 wk, respectively). Septa with 1,000 microg per septum had a significant decline in the rate of pheromone release for this 4 wk exposure time (total = 119 microg/4 wk). A large proprietary wax granule (44 mg per granule, 25% wt:wt pheromone) provided a steady rate of pheromone release (total = 2,347 microg/4 wk per granule). Experimental granules (16 mg per granule) made of soywax with higher pheromone loads (10% wt:wt) approached zero order release (steady state) (total = 69 microg/4 wk per granule), whereas smaller granules (4 mg per granule) with less pheromone (0.1% wt:wt) provided first-order release profiles (decreasing rate with longer exposure time) (total = 0.35 microg/4 wk per granule). A field trial in turfgrass demonstrated the potential of selected granular formulations to provide effective mating disruption for up to 4 wk, as measured by pheromone trap shutdown. Documenting pheromone release profiles for these experimental granules and rubber septa provides valuable information that will support future field evaluations of mating disruption as a control strategy. PMID- 19133468 TI - Impact of the zoophytophagous plant bug Nesidiocoris tenuis (Heteroptera: Miridae) on tomato yield. AB - Nesidiocoris tenuis Reuter (Heteroptera: Miridae) is one of the most controversial dicyphines because of the injuries it causes to tomato (Solanum esculentum) crops. The aim of this study was to quantify the impact of N. tenuis on fruit yield. Tomato plants were exposed to an average of 0.53 +/- 0.26, 3.4 +/ 1.1, 12.0 +/- 1.4, and 35.2 +/- 7.7 N. tenuis per plant for 3 wk. When fruit were exposed to N. tenuis as flowers, the percentage of aborted fruit was higher in compartments infested with N. tenuis, and this was related directly to the cumulative number of N. tenuis (CNN). However, compartments with the highest abortion rates had heavier and bigger fruit and were not significantly different from the controls. The variation in fruit weight was satisfactorily explained with a logistic equation in relation to the proportion of aborted fruit. To estimate the density thresholds, the variation in truss weight was predicted as a function of fruit weight and aborted fruit. The two latter variables were expressed as a function of the CNN. A maximum of 15% truss weight overcompensation was predicted at 15.8% of fruit abortion. Yield reduction was predicted at fruit abortion rates > or = 27.7%, which corresponded to 566 CNN per plant or 32.11 CNN per leaf. N. tenuis may be considered a useful predator of small pests in tomato crops if kept under these thresholds. Mathematical models predict a yield increase and fruit upgrade that overcompensates for the reduction in the number of fruit below the density threshold. PMID- 19133469 TI - Effects of a killed-cover crop mulching system on sweetpotato production, soil pests, and insect predators in South Carolina. AB - Sweetpotatoes, Ipomoea batatas (L.) Lam. (Convolvulaceae), are typically grown on bare soil where weeds and erosion can be serious problems. Conservation tillage systems using cover crop residues as mulch can help reduce these problems, but little is known about how conservation tillage affects yield and quality of sweetpotato or how these systems impact populations of beneficial and pest insects. Therefore, field experiments were conducted at the U.S. Vegetable Laboratory, Charleston, SC, in 2002-2004 to evaluate production of sweetpotatoes in conventional tillage versus a conservation tillage system by using an oat (Avena sativa L. (Poaceae)-crimson clover (Trifolium incarnatum L.) (Fabaceae) killed-cover crop (KCC) mulch. The four main treatments were 1) conventional tillage, hand-weeded; 2) KCC, hand-weeded; 3) conventional tillage, weedy; and 4) KCC, weedy. Each main plot was divided into three subplots, whose treatments were sweetpotato genotypes: 'Ruddy', which is resistant to soil insect pests; and 'SC1149-19' and 'Beauregard', which are susceptible to soil insect pests. For both the KCC and conventional tillage systems, sweetpotato yields were higher in plots that received hand weeding than in weedy plots. Orthogonal contrasts revealed a significant effect of tillage treatment (conventional tillage versus KCC) on yield in two of the 3 yr. Ruddy remained resistant to injury by soil insect pests in both cropping systems; and it consistently had significantly higher percentages of clean roots and less damage by wireworm-Diabrotica-Systena complex, sweetpotato flea beetles, grubs, and sweetpotato weevils than the two susceptible genotypes. In general, injury to sweetpotato roots by soil insect pests was not significantly higher in the KCC plots than in the conventionally tilled plots. Also, more fire ants, rove beetles, and carabid beetle were captured by pitfall traps in the KCC plots than in the conventional tillage plots during at least 1 yr of the study. This study suggests that a sweetpotatoes can be successfully grown under a killed-cover crop production system. PMID- 19133470 TI - Relative repellency and lethality of the neonicotinoids thiamethoxam and acetamiprid and an acetamiprid/bifenthrin combination to Reticulitermes flavipes termites. AB - Field-collected Reticulitermes flavipes (Kollar) termites were placed in bioassay tubes containing soil treated with one of three termiticides: thiamethoxam, acetamiprid, or a combination of acetamiprid + bifenthrin. In the bioassay tubes, treated soil was placed in a layer centered within untreated sand between two 1.5 cm agar plugs. All termiticides were tested at concentrations of 0.1, 1, 10, and 100 ppm with narrow (1 mm), medium (5 mm), and broad (50 mm) thicknesses of treated soil. Soil penetration and termite mortality were measured after 7 d, and repellency was assessed. Thiamethoxam treatments allowed the greatest soil penetration, whereas acetamiprid + bifenthrin treatments were the most inhibitory to soil penetration. Thiamethoxam treatments also caused consistently greater termite mortality than acetamiprid treatments. These data indicated that acetamiprid prevented soil penetration by termites more than thiamethoxam, although both were less repellent compared with bifenthrin alone, which causes little termite mortality at the tested doses. When there was direct contact of treated soil with the agar plugs in broad treatments, the combination of acetamiprid + bifenthrin was more toxic to R. flavipes termites than either acetamiprid or thiamethoxam alone. The combination acetamiprid + bifenthrin termiticide may be effective in keeping termites away from the treated soil, because of the combined effects of acetamiprid and bifenthrin. PMID- 19133471 TI - Cloning and characterization of a GABA receptor from Plutella xylostella (Lepidoptera: Plutellidae). AB - A full-length cDNA, with an open reading frame (ORF) of 1,449 bp, encoding a subunit of the gamma-aminobutyric acid (GABA)-activated chloride channel was isolated from Plutella xylostella (L.) (Lepidoptera: Plutellidae) (GenBank accession no. EF156251). The subunit gene encoded a 483-amino acid polypeptide that showed 84% sequence identity with DmRdl subunit (U02042) (Drosophila melanogaster resistant to dieldrin). When expressed in Xenopus laevis oocytes, the subunit assembled as a functional homomeric complex activated by GABA and abamectin in a dose-dependent manner. The EC50 value of GABA was 0.49 mM (0.41 0.58) (n = 5). However, the responses to abamectin were very robust, with an EC50 of 4.85 microM (4.02-5.89) (n = 6), indicating that abamectin was > 100-fold more potent in activating chloride currents than GABA. The results suggest that this subunit is vital to the formation of a functional channel and contains the binding site of abamectin. PMID- 19133472 TI - Dynamics of esterase alleles in Culex pipiens complex mosquitoes in Beijing. AB - To investigate insecticide resistance levels and dynamic changes of carboxylesterase polymorphism with time in resistant populations of mosquitoes in the Culex pipiens complex, four field populations were collected in September 2006. The resistance levels of fourth-instar larvae to organophosphate (dichlorvos, parathion, and chlorpyrifos), carbamate (fenobucarb and propoxur), and pyrethroid (permethrin and tetramethrin) insecticides were determined by bioassay. Larvae had a low but significant resistance to organophosphate and carbamate insecticides but no significant resistance to pyrethroid insecticides. Starch gel electrophoresis revealed the presence of the overproduced esterases B1, A2-B2, A8-B8, A9-B9, and All-B11. The frequency of each overproduced esterases varied depending on its regional localities. Compared with previous surveys, the polymorphism of amplified esterase alleles in Beijing populations increased, although the resistance level to organophosphate insecticides declined. PMID- 19133473 TI - Selection and life history traits of tebufenozide-resistant sugarcane borer (Lepidoptera: Crambidae). AB - Varying susceptibility to tebufenozide was recorded in the sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), collected from Louisiana sugarcane locations with different selection pressures. Results from diet incorporation bioassays with tebufenozide indicated significant increases in LC50 (3.78-fold) and LC90 (7-fold) values for a colony from Duson (DU), an area with higher selection pressure, compared with a colony from Alexandria (ALEX), an area with no selection pressure. Differences were not detected in LC50 values among colonies from areas where use of tebufenozide was discontinued or rotated with other chemistries. Selections with tebufenozide of DU larvae over 12 generations resulted in a highly resistant colony (DU-R) with 27.1- and 83.3-fold increases in LC50 and LC90 values, respectively. Comparison of pupal weight, days to pupation, and emergence after exposure to an equitoxic (LC20) concentration of tebufenozide revealed a decrease in pupal weight (34 and 33% for males and females, respectively), and an increase in days to pupation (47 and 40% for males and females, respectively), and emergence (43 and 33% for males and females, respectively) for the DU-R colony compared with the parent DU colony. Fecundity of DU-R females decreased to 72 eggs per female compared with 180 (DU) and 261 (ALEX). Egg viability of the ALEX and DU colonies was 61 and 56%, respectively, whereas only 27% of eggs laid by females from the DU-R colony hatched. These results are discussed in terms of their practical implications for control of D. saccharalis in Louisiana sugarcane. PMID- 19133474 TI - Genetic characterization of resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from India. AB - The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is considered as one of the most difficult pests to control. It has developed resistance not only to synthetic insecticides but also to Bacillus thuringiensis based pesticides. We tested the hypothesis that selection in a P. xylostella population, from Hosur, India, with deltamethrin would give a broad spectrum of resistance to several insecticides. We also were interested in genetically classifying resistance to deltamethrin in the selected population and in evaluating whether resistance can be suppressed using synergists. Bioassays (at generation 1, G1) using deltamethrin indicated a resistance ratio of 161-fold compared with a laboratory-susceptible population of P. xylostella (Lab-UK). At G2, the field-derived population was divided into two subpopulations; one population was selected (G1 to G8) with deltamethrin (Delta-SEL), and the second population was left unselected (UNSEL). Bioassays at G9 indicated that selection with deltamethrin gave a resistance ratio of 15-fold compared with UNSEL and 1,647-fold compared with Lab-UK. The resistance to deltamethrin in the UNSEL population was stable. The Delta-SEL population maintained resistance to lambda cyhalothrin, but there was no cross-resistance to indoxacarb, DDT, or Cry1Ac. Crossing experiments indicated that resistance to deltamethrin in Delta-SEL was multigenic and inherited in an incompletely dominant fashion. Piperonyl butoxide (PBO) and S.S.S-tri-n-butyl phosphorotrithioate with potent inhibitory activity against esterases and/or monooxygenases significantly increased the toxicity of deltamethrin against both UNSEL and Delta-SEL, but they showed no such synergism with Lab-UK. Thus, it can be predicted that development of resistance to deltamethrin would be delayed under appropriate control strategies that favor the dilution of resistance alleles by enhanced flow of susceptible alleles. Further analysis suggested that mixing PBO and deltamethrin could eliminate the substantial resistance to deltamethrin in this population. PMID- 19133475 TI - Survey for resistance to four insecticides in Myzus persicae clones from peach trees and weeds in south-central Washington. AB - The insecticides esfenvalerate, endosulfan, imidacloprid, and methamidophos were screened against 74 clonal populations of the green peach aphid, Myzus persicae (Sulzer), collected as fundatrices from peach (Prunus spp.) and as apterous virginoparae from various weeds near potato (Solanum spp.) fields in the Yakima Valley and Columbia Basin regions of Washington state. Response to diagnostic concentration of four insecticide products demonstrated a bimodal pattern of survival to endosulfan (Phaser), suggesting two phenotype classes responding to this insecticide. This pattern was not observed with the other products. Moderately significant correlations in mortality of aphid clones treated with diagnostic concentrations of endosulfan versus clones treated with methamidophos and similarly with the correlation of mortalities for imidacloprid-treated versus methamidophos-treated clones suggested modest levels of cross tolerance. No significant correlations were observed with the remaining four possible comparisons. Concentration-response bioassays were conducted on 16 clones with the four insecticides. The greatest difference between resistant and susceptible clones (expressed as the ratio of lethal concentrations producing 50% mortality; RR) was only seven-fold observed in the endosulfan-treated clones. The greatest RR was five-fold for imidacloprid, four-fold for esfenvalerate, and three-fold for methamidophos-treated clones. Only the endosulfan response is likely to be ofbiological significance and reflects the same cyclodiene resistance discovered in this region over a decade ago. PMID- 19133476 TI - Fitness costs limit the development of resistance to indoxacarb and deltamethrin in Heliothis virescens (Lepidoptera: Noctuidae). AB - Insecticide resistance in Heliothis virescens (F.) (Lepidoptera: Noctuidae) has been documented from all over the world and is often associated with reduced fitness. Fitness costs could delay the development of resistance depending upon the prevailing conditions. We were interested in establishing whether a field collected population from Washington County, MS, was resistant to spinosad, indoxacarb, and deltamethrin and whether any such resistance was associated with fitness costs. Bioassays results showed that the insecticides were equally toxic to the field population. Upon laboratory selection (generations [G]3 to G8), the resistance ratio increased only 2-, 3-, and 1-fold for spinosad, deltamethrin, and indoxacarb, respectively, compared with the field population. In contrast, the resistance ratios increased 213-, 65-, and 55-fold compared with an unselected population at G9. The estimated realized heritability (h2) after six generations of selection was 0.17, 0.03, and 0.12, respectively, and the number of generations required for 10-fold increase in LC50 of Spino-SEL, Indoxa-SEL, and Delta-SEL was estimated to be 14.3, 50, and 14.3. Comparison of life traits between the selected and unselected populations revealed that the selected populations laid a significantly lower number of eggs and that a lower percentage of eggs hatched. This also was reflected in both the net replacement rate and the intrinsic rate of population increase, which were both lower for the selected populations. It also was observed that the mean relative growth rate of the larvae was lower for the selected populations; not only did the larvae take longer to pupate but the mean weight of the prepupae from the selected populations was lower. Our data suggest that due to fitness costs the development of resistance to the insecticides was limited such that after six generations of selection the larvae were no less susceptible to the insecticides than the field population although were considerable more resistant than the unselected population. PMID- 19133477 TI - Developing and testing a diagnostic probe for grape phylloxera applicable to soil samples. AB - Grape phylloxera, Daktulosphaira vitifoliae (Fitch) (Hemiptera Phylloxeridae) is a damaging pest of grapevines (Vitis spp.) around the world, and the management of this pest requires early detection of infestations. Here, we describe the development and validation of a sensitive DNA test for grape phylloxera that can be applied to soil. Species-specific primers were developed for grape phylloxera in the internal transcribed space region 2, and their specificity was confirmed after thorough screening by using a wide range of vineyard organisms and aphid genera. Preliminary testing of the detection limits of the grape phylloxera specific primers was conducted using field-sourced soil types spiked with a known number of grape phylloxera. The assay was converted to a real-time polymerase chain reaction format (TaqMan MGB). This assay, in combination with DNA extraction from soil, can detect phylloxera crawlers added to soil. The assay was evaluated in the field at a recently detected grape phylloxera infestation site from the Yarra Valley in Victoria, Australia. The DNA assay proved to be substantially more sensitive than a standard ground survey for detecting grape phylloxera presence on vine roots in the infested vineyard. Moreover, unlike the ground survey, the assay provided quantitative information on grape phylloxera infestations, because grape phylloxera DNA concentrations in samples from vines closely matched the numbers of grape phylloxera crawlers collected with emergence traps placed at the base of vines. Unlike other detection techniques, the method can be applied at any time of the year, and it can be potentially modified to provide specific information on the virulence levels of the particular grape phylloxera genotypes responsible for any new infestations. PMID- 19133478 TI - DNA diagnostics of three armored scale species on kiwifruit in New Zealand. AB - Three species of armored scale (Hemiptera: Diaspididae) are found on kiwifruit (Actinidia spp.) in New Zealand orchards: latania scale, Hemiberlesia lataniae (Signoret); greedy scale, Hemiberlesia rapax (Comstock); and oleander scale, Aspidiotus nerii (Bouche). Each of them is a quarantine pest in some of the markets to which New Zealand kiwifruit are exported. Adult females of the three species can be distinguished morphologically; however, the task is laborious when large numbers must be identified. Furthermore, it is not possible to distinguish among the immature stages. A DNA-based diagnostic using a multiplex polymerase chain reaction (PCR) method based on differences in the cytochrome oxidase I and II genes was developed to distinguish the three species. The test relies on the rapid isolation of amplifiable DNA by using a protease (prepGEM), followed by multiplex PCR using primers that distinguish the species at three or more nucleotide positions within cytochrome oxidase I and II, resulting in PCR products of characteristic size for each species. The test was validated in a double-blind experiment and then used to determine the relative distribution and abundance of the three species on leaves and fruit of 'Hayward' and 'Hortl6A' kiwifruit across the dominant growing regions throughout New Zealand during the 2007 season. In total, 3,418 scale insects were identified to species level: 1,904 (56%) were latania scale; 1,473 (43%) were greedy scale; and 41 (1%) were oleander scale. Since the last survey in 1988, latania scale has displaced greedy scale as the dominant species of armored scale on Hayward kiwifruit in the North Island and was found for the first time in the South Island. Only a single latania scale was found on Hortl6A fruit, consistent with previous reports of reduced rates of settlement on the fruit of this cultivar by latania scale compared with greedy scale. PMID- 19133479 TI - Efficacy of transgenic cotton expressing Cry1Ac and Cry1F insecticidal protein against heliothines (Lepidoptera: Noctuidae). AB - Cotton, Cossypium hirsutum L, plants expressing Cry1Ac and Cry1F (Phytogen 440W) insecticidal crystal proteins of Bacillus thuringiensis (Bt) Berliner, were evaluated against natural populations of tobacco budworm, Heliothis virescens (F.), and bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), across 13 southern U.S. locations that sustained low, moderate, and high infestations. The intrinsic activity of Phytogen 440W was compared with nontreated non-Bt cotton (PSC355) and with management strategies in which supplemental insecticides targeting heliothines were applied to Phytogen 440W and to PSC355 cotton. Infestations were composed primarily of bollworm, which is the least sensitive of the heliothine complex to Cry toxins. Therefore, damage recorded in these studies was primarily due to bollworm. Greater than 75% of all test sites sustained heliothine infestations categorized as moderate to high (10.6-64.0% peak damaged bolls in nontreated PSC355). Phytogen 440W, alone or managed with supplemental insecticide applications, reduced heliothine-damaged plant terminals, squares (flower buds), flowers, and bolls equal to or better (1.0-79.0-fold) than managing a non-Bt cotton variety with foliar insecticides across all infestation environments. Rarely (frequency of < or = 11% averaged across structures), sprayed Phytogen 440W reduced damaged structures compared with nontreated Phytogen 440W. Protection against heliothine-induced plant damage was similar across the three levels of infestation for each viable management strategy, with exception to damaged squares for nontreated Phytogen 440W. In situations of moderate to high heliothine infestations, cotton plants expressing Cry1Ac and Cry1F may sustain higher levels of damage compared with that same variety in low infestations. No significant difference in yield was observed among heliothine management strategies within each infestation level, indicating cotton plants may compensate for those levels of plant damage. These findings indicate Phytogen 440W containing Cry1Ac and Cry1F provided consistent control of heliothines across a range of environments and infestation levels. PMID- 19133480 TI - Toxicity of cassia and cinnamon oil compounds and cinnamaldehyde-related compounds to Sitophilus oryzae (Coleoptera: Curculionidae). AB - The toxicity to adult Sitophilus oryzae (L.) (Coleoptera: Curculionidae) of two cassia oils (Especial and true), four cinnamon oils (technical, #500, bark, and green leaf), and (E)-cinnamaldehyde and its 41 structurally related compounds was examined by residual and vapor-phase toxicity bioassays. Results were compared with those of dichlorvos. In residual bioassays, cassia and cinnamon oils exhibited good insecticidal activity. Based on 48-h LD50 values, the toxicity of allyl cinnmate (0.0003 mg/cm2) was comparable with that of dichlorvos (0.00025 mg/cm2). Potent insecticidal activity also was observed with benzaldehyde, beta caryophyllene, cinnamonitrile, hydrocinnamyl acetate, (E)-4-hydroxycinnamic acid, and alpha-terpineol (LD50 = 0.003-0.009 mg/cm2). Structure-activity relationships indicate that types of functional groups rather than hydrophobicity or vapor pressure parameters seem to play a role in determining the toxicities to adult S. oryzae. In vapor-phase toxicity tests with weevils, these compounds were more effective in closed containers than in open ones. These results indicate that the effect of the compounds was largely a result of action in the vapor phase. Cassia and cinnamon oils and test compounds described merit further study as potential fumigants for the control of S. oryzae because of their greater activity as a fumigant. PMID- 19133481 TI - Infestation and population dynamics of insects on stored cassava and yams chips in Benin, West Africa. AB - Natural insect infestation in cassava (Manihot esculenta Crantz subspecies esculenta) and yam (Dioscorea spp.) chips was evaluated during two consecutive storage seasons (2003-2004 and 2004-2005) in two agroecological zones of Benin (Northern Guinea Savanna [NGS] and Sudan Savanna [SS]). The insects infesting chips were collected, identified, and counted, they included Prostephanus truncatus (Horn) (Coleoptera: Bostrychidae), Cathartus quadricollis (Guerin) (Coleoptera: Silvanidae), Carpophilus dimidiatus (F.) (Coleoptera: Nitidulidae), Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), and Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae). P. truncatus and C. quadricollis were observed with a higher prevalence on cassava than on yam chips. During both seasons after 3 mo of storage, all (100%) cassava chip samples were infested with P. truncatus and C. quadricollis in both agroecological zones, whereas yam chips only showed lower infestation rates of 59.5 and 19.1% for P. truncatus and C. quadricollis, respectively, at the end of storage in 2003-2004. During the 2004 2005 season after 3 mo of storage infestation rate in yam chips was 66 and 24% in NGS and 100 and 0% in SS for P. truncatus and C. quadricollis, respectively, showing that insect infestation levels vary significantly with commodity, year, and fluctuate during the storage season. PMID- 19133482 TI - Differential heat shock tolerance and expression of heat-inducible proteins in two stored-product psocids. AB - The recent recognition of psocids as a major concern in stored products and also the reemergence of heat treatment as a control tactic of stored-product insects led to the present investigation. The objectives of this study were to determine whether there are differences in heat shock tolerance of two species of stored product psocids--Lepinotus reticulatus Enderlein (Trogiidae) and Liposcelis entomophila (Enderlein) (Liposcelididae)--and to determine whether heat shock proteins (HSPs) underlay such tolerance. Time-response bioassays were therefore carried out at increasing temperatures for both psocids. The lethal time (LT)50 and LT95 estimates were correlated with the expression of heat shock proteins after exposure at the same range of temperatures for 30 min. The expression of HSP was determined through Western blot analyses using HSP 70 antibody. Liposcelis entomophila was more than two-fold more tolerant than L. reticulatus for nearly all of the range of temperatures (> or = 40.0 degrees C). Expression of HSP 70 was not observed for either of the psocid species, but the expression of two low-molecular-mass heat-inducible proteins (HIPs; 23 and 27 kDa) was observed in L. entomophila. The expression of these small proteins was induced by exposure to higher temperatures, and the trend was particularly strong for HIP 27. In contrast, no expression of small heat-inducible proteins was detected in L. reticulatus, reflecting its higher susceptibility to heat treatments. The relatively high heat tolerance of L. entomophila might help explain its more common occurrence in grain stored in warmer regions of the world. PMID- 19133483 TI - Effect of systemic intermittent administration of human parathyroid hormone (rhPTH[1-34]) on the resistance to reverse torque in rabbit tibiae. AB - The aim of this study was to evaluate the effect of intermittent administration of human parathyroid hormone [rhPTH (1-34)] on the removal torque of implants placed in rabbit tibiae. Twenty male New Zealand rabbits were submitted to implant surgery. Each animal received one machined screw-type implant (3.75 mm diameter x 8 mm length) in the proximal metaphysis of the right tibia. The rabbits were then divided into 2 groups: the test group (n = 10) received 6 microg/kg of rhPTH (1-34) subcutaneously in the dorsal region 3 days a week, and the control group (n = 10) received placebo. Removal torque was performed at 28 and 56 days after implant placement for both groups. The mean removal torque values at 28 days were 37.0 +/- 4.36 Ncm and 47.4 +/- 6.77 Ncm for control and test groups respectively (P < .05). At 56 days the reverse torque was 45.8 +/- 3.96 Ncm for the control group and 55.8 +/- 2.86 Ncm for the test group, indicating that the removal torque was significantly higher in the test groups (P < .05). These results demonstrated that intermittent treatment with rhPTH (1-34) enhanced the removal torque of implants in rabbit tibiae. PMID- 19133485 TI - Vertical alveolar ridge expansion and simultaneous implant placement in posterior maxilla using segmental osteotomy: report of two cases. AB - Implant placement in vertically insufficient alveolar ridges of the posterior maxillae may require bone augmentation, such as sinus floor elevation, onlay grafting, and distraction osteogenesis. The purpose of this report is to evaluate the feasibility of vertical alveolar ridge expansion and simultaneous implant placement in the posterior maxilla using segmental osteotomy without sinus floor elevation, onlay grafting, and distraction osteogenesis. Two patients presenting with vertically insufficient alveolar ridges in the posterior maxillae were treated with the surgical procedure as follows. The edentulous ridge was split vertically using a segmental osteotomy before the preparation of an implant bed, and 1 implant body was placed simultaneously in the ridge with the vertical split. Three months later, abutments were connected and prosthetic loading of the implants was started. The follow-up was performed with the aid of dental and panoramic radiographs, evaluation of peri-implant probing depths, and Periotest values. In both patients, alveolar ridge expansion and implant placement were carried out without problems, good functional results were performed, and the clinical and radiologic findings were satisfactory. The outcome of the surgical procedure showed minimal invasion and good functional results. PMID- 19133484 TI - Arachidonic acid and prostaglandin E2 influence human osteoblast (MG63) response to titanium surface roughness. AB - Prior studies have shown that implant surface roughness affects osteoblast proliferation, differentiation, matrix synthesis, and local factor production. Further, cell response is modulated by systemic factors, such as 1,25(OH)2D3 and estrogen as well as mechanical forces. Based on the fact that peri-implant bone healing occurs in a site containing elevated amounts of prostaglandin E2 (PGE2), the hypothesis of the current study is that PGE2 and arachidonic acid (AA), the substrate used by cyclooxygenase to form PGE2, influence osteoblast response to implant surface roughness. To test this hypothesis, 4 different types of commercially pure titanium (cpTi) disks with surfaces of varying roughness (smooth Ti, R(a) 0.30 microm; smooth and acid etched Ti [SAE Ti], R(a) 0.40 microm; rough Ti, R(a) 4.3 microm; rough and acid etched Ti [RAE Ti], R(a) 4.15 (microm) were prepared. MG63 osteoblasts were seeded onto the surfaces, cultured to confluence, and then treated for the last 24 hours of culture with AA (0, 0.1, 1, and 10 nM), PGE2 (0, 1, 10, 25, and 100 nM), or the general cyclooxygenase inhibitor indomethacin (0 or 100 nM). At harvest, the effect of treatment on cell proliferation was assessed by measuring cell number and [3H]-thymidine incorporation, and the effect on cell differentiation was determined by measuring alkaline phosphatase (ALP) specific activity. The effect of AA and PGE2 on cell number was somewhat variable but showed a general decrease on plastic and smooth surfaces and an increase on rough surfaces. In contrast, [3H]-thymidine incorporation was uniformly decreased with treatment on all surfaces. ALP demonstrated the most prominent effect of treatment. On smooth surfaces, AA and PGE2 dose-dependently increased ALP, while on rough surfaces, treatment dose dependently decreased enzyme specific activity. Indomethacin treatment had either no effect or a slightly inhibitory effect on [3H]-thymidine incorporation on all surfaces. In contrast, indomethacin inhibited ALP on smooth surfaces and stimulated ALP on rough. Taken together, the results indicate that both AA and PGE2 influence osteoblast response by promoting osteoblast differentiation on smooth surfaces, while inhibiting it on rough surfaces. Because implants with rough surfaces are acknowledged to be superior to those with smooth surfaces, these results suggest that use of nonsterioidal anti-inflammatory drugs to block PGE2 production and reduce inflammation may be beneficial in the postoperative period after implant placement. They also indicate that manipulation of the AA metabolic pathway may offer a new therapeutic approach for modulating bone healing after implant placement. Because peri-implant healing takes place in a complex cellular environment quite different from the one used in the present study, additional work will be necessary to substantiate these possibilities. PMID- 19133486 TI - A modified crestal ridge expansion technique for immediate placement of implants: a report of three cases. AB - Atrophic edentulous jaws can pose a significant challenge to successful oral rehabilitation with endosseous dental implants. Although ridge augmentation can help to restore ridge volume, grafting procedures can significantly increase patient morbidity, costs, and treatment time, depending on the case, before dental implants can be placed. This article reports on an alternative technique used in 3 patients to expand ridge volume and place dental implants in a single procedure. A partial-thickness flap was elevated to expose the alveolar crest, and conventional implant osteotomies were partially prepared. Along the crest of the ridge, a furrow with terminal vertical releases 1 to 3 mm deep were created, and a bone chisel was used to deepen the furrow. Osteotomes were used to complete preparation of the implant receptor sites, and the implants were placed. Bony plates were stabilized through the use of resorbable sutures. Furrows more than 2 mm deep between the plates were augmented with a xenograft. Collegen membranes were placed over the sites, and soft tissue was sutured. Healing was unremarkable, and all implants were successfully restored. For these patients, the ridge expansion technique resulted in substantial bone reconstruction with little or no grafting. Long-term, prospective studies on this procedure are required before definitive conclusions can be drawn. PMID- 19133487 TI - A new twist on surgical guides. AB - Surgical guides have evolved to the point that they can be created with the aid of computer-assisted tomography (CT) technology. Surgical guides also are used in construction of the prosthesis prior to implant surgery. With existing guide techniques, the prosthesis is screw retained, or cone-shaped temporary abutments may be used to overcome the lack of parallelism of the implants. With the New Twist technique, we are able to create the surgical guide in such a way that optimum bone may be used and the rotational position of the implant may be established for the surgeon and for the laboratory. Custom abutments are inserted at the time of surgery, as is an esthetic, functionally provisional cemented prosthesis. Castings for the finished ceramo-metal bridge are fabricated before surgery is performed. The final prosthesis then is easily constructed by the restorative dentist during just 3 simple patient visits. PMID- 19133488 TI - Repair of the perforated sinus membrane with buccal fat pad during sinus augmentation. AB - Several reports demonstrate successful use of the buccal fat pad (BFP) as pedicled graft in reconstructing small to medium sized maxillary defects. BFP harvesting has so far been shown to be an easy, well-tolerated, and uncomplicated technique for oral reconstruction. This case report proposes the use of BFP for repairing of the perforated sinus membrane during sinus augmentation. PMID- 19133489 TI - Clinical and histomorphometric evaluation of staged approach using bone augmentation and autogenous masticatory mucosal graft with endosseous implant placement: a case report. AB - Various materials have been used to prevent or minimize ridge collapse after tooth extraction in an attempt to improve implant placement. However, there is no report evaluating the effect the additional autogenous masticatory mucosal graft (free gingival graft [FGG]) on the maturation of the grafted area. A patient who presented with a missing area with prior bone-augmentation procedure was treated with FGG. The additional procedure of FGG performed well as a soft tissue barrier for frenal pull, and the underlying bone showed good healing maturity with a high percentage of new bone formation. PMID- 19133490 TI - Psychiatric nursing as gift giving. PMID- 19133491 TI - Understanding the clinical profile of a drug on the basis of its pharmacology: mirtazapine as an example. AB - Direct comparisons of different drugs used for the same indication can reveal important differences in efficacy or tolerability. Such clinical differences can often be explained or predicted on the basis of the particular pharmacological properties of each drug. To demonstrate the clinical importance of understanding the distinct pharmacology of a drug, this article describes the pharmacology and clinical profile of mirtazapine (Remeron). Mirtazapine is a pharmacologically unique agent among antidepressant drugs. Clinical studies have demonstrated similar overall efficacy for mirtazapine compared with the serotonin reuptake inhibitors and with venlafaxine (Effexor), but a more rapid onset of antidepressant effect and a substantially different side effect profile. Knowledge about drug pharmacology leads to better understanding of the differences in the efficacy and tolerability of various drugs. This knowledge also provides a rationale for choosing one drug over another for a particular patient or switching or combining drugs for treating nonresponders or alleviating side effects. PMID- 19133492 TI - A creative arts intervention for children with diabetes. Part 2: evaluation. AB - In Part 1 of this article (published in the October 2008 issue), we discussed the importance of using creative arts skits as an expressive technique for children with Type 1, or juvenile, diabetes. This creative arts intervention offers children the opportunity to decipher emotional difficulties through symbolic play in a secure atmosphere. Analysis of feelings following the skits encourages children to share concerns about their illnesses as well as self-concept issues. In Part 2, we use the case study method to demonstrate the benefits of creative arts skits for children with diabetes. PMID- 19133493 TI - Equine-facilitated psychotherapy with adult female survivors of abuse. AB - This qualitative study examined the stories of 5 women who experienced abuse and participated in equine-facilitated psychotherapy (EFP) as part of their recovery. Anecdotal accounts support the effectiveness of EFP with women who have experienced abuse, but there is a lack of supporting research. This study was designed to examine the effectiveness of EFP in the treatment of women who have experienced abuse. Selection criteria included age, experience of abuse, participation in EFP, and ability to understand English. Data analysis identified four patterns in the participants' stories: I Can Have Power; Doing It Hands On, Horses as Co-Therapists, and Turned My Life Around. Overall, the participants' stories show that EFP can be an effective intervention for women who have experienced abuse. PMID- 19133494 TI - Promoting dual diagnosis awareness in everyday clinical practice. AB - Working with and treating individuals with co-occurring substance misuse presents unique challenges for mental health nurses. Substance misuse is common in mental health settings, and its clinical challenges with system barriers impede positive treatment outcomes. To work effectively in everyday clinical practice with dual diagnosis clients, nurses need to consider potential guiding principles. These principles pertain to treatment aims, engagement, assessment, symptoms, care plans, cycle of change, motivational interviewing, traps to avoid, active treatment, relapse prevention, agency collaboration, and treatment diversity. Clinical tools are easily accessible resources that can provide a framework for contemporary practice in a range of settings. Such resources are helpful in practice and as a foundation for orientation, education, self-reflection, and peer review. Setting expectations for everyday practice and ensuring that clients with a dual diagnosis receive the best care possible are important goals for the nursing profession. PMID- 19133495 TI - A model of CAM use by women with female-specific cancers. AB - In this cross-sectional, retrospective, explanatory secondary analysis of the 2002 National Health Interview Survey, complementary and alternative modality (CAM) use by women with female-specific cancers is described. The effect of personal factors on the odds of CAM use was measured using the researcher's model, based on Pender's Health Promotion Model, as a framework. Women age 40 and older, with high levels of edu cation, presence of pain, and presence of depression/anxiety were more likely to use CAM. Because women with female specific cancers who report depression/anxiety are more likely to use CAM, they need to be targeted by mental health nurses for education regarding safe CAM use. PMID- 19133496 TI - Biological activity and identification of neuropeptides in the neurosecretory complexes of the cabbage pest insect, Mamestra brassicae (Noctuidae; Lepidoptera). AB - The need for more environmentally sound strategies of plant protection has become a driving force in physiological entomology to combat insect pests more efficiently. Since neuropeptides regulate key biological processes, these "special agents" or their synthetic analogues, mimetics, agonists or antagonists may be useful tools. We examined brain-suboesophageal ganglia and corpora cardiaca-corpora allata complexes of the cabbage moth, Mamestra brassicae, in order to obtain clues about possible peptide candidates which may be appropriate for the biological control of this pest. With the aid of bioassays, reversed phase high performance liquid chromatography, and mass spectrometry, five neuropeptides were unequivocally identified and the presence of a further three were inferred solely by comparing mass spectra with known peptides. Only one neuropeptide with adipokinetic capability was identified in M. brassicae. Data from the established homologous bioassay indicated that the cabbage moths rely on a lipid-based metabolism which is aided by an adipokinetic hormone (viz. Manse AKH) that had previously been isolated in many different lepidopterans. Other groups of neuropeptides identified in this study are: FLRFamides, corazonin, allatostatin and pheromonotropic peptide. PMID- 19133497 TI - Thyroidal inhibition following diverse stress in soft-shelled turtle, Lissemys punctata punctata bonnoterre. AB - The current study was undertaken to ascertain the effects of diverse stress on thyroid activity in soft-shelled turtles, Lissemys punctata punctata. The findings revealed that starvation (10 days), dehydration (10 days) or exposure to electric shock (12 volts for 15 seconds at an interval of 30 min for 3 h) caused significant decrease in the body weight (except in electric shock), relative weight, peripheral and central epithelial heights of the follicles and peroxidase activity of the thyroid gland of turtles. The degree of change in the values of these parameters was nearly same in all the stress experiments, indicating that there is not much difference in the degree of thyroid responses to diverse stress in turtles. It is suggested that these stressors might have exerted their actions on thyroid activity presumably indirectly via adrenal medulla and/or substance in metabolic stress (starvation and dehydration) and via hypothalamo-hypophysial adrenocortical axis in non-metabolic stress (electric shock) in Lissemys turtles. PMID- 19133498 TI - Rat testicular mitochondrial antioxidant defence system and its modulation by aging. AB - Accumulation of oxidative damage caused by reactive oxygen species (ROS) underlies fundamental changes found during aging. In the present study, age related effect on testicular mitochondrial oxidant generation and antioxidant defence profile was investigated in Wistar rats at 3 months (young adults), 12 months (old adults) and 24 months (senescent animals) of age. Mitochondrial oxidative stress parameters viz., lipid peroxidation (LPx), protein carbonylation, hydrogen peroxide (H2O2) generation and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), levels of total, oxidized (GSSG) and reduced glutathione (GSH) were studied to find out their roles in maintenance of mitochondrial glutathione redox pool as a function of age. Increased levels of LPx, H2O2 and decreased GSH content accompanied by a decline in activities of SOD, GPx and GR with advancing age suggest that antioxidant defense profile of testicular mitochondria exhibit age related alterations which might play a critical role in regulating physiological functions of the testis such as steroidogenesis and spermatogenesis. PMID- 19133499 TI - Investigation of abiogenic stress-induced alterations in the level of secondary metabolites in poppy plants (Papaver somniferum L.). AB - We aimed to understand the effects of water stress on the alkaloid production in various developmental stages of poppy plants and the effect of stress on the alkaloids content in the capsules. Three stages of the life cycle of Papaver somniferum L. were selected in our studies: Rosette, Flowering and Lancing developmental stages. Four types of water conditions were examined: Control, Withdrawal of Water, 50% Water Supply and Inundation. The morphological monitoring, results of Relative Water Content and proline content were used as indicators of stress. The result of the measurements in poppy leaves show that the secondary metabolites dramatically respond to these stress conditions. The constant water supply was beneficial for the accumulation of alkaloids in the capsules. PMID- 19133500 TI - Modeling and biochemical analysis of the activity of antibiofilm agent Dispersin B. AB - Bacteria in a biofilm are enmeshed in a self-synthesized extracellular polysaccharide matrix (PGA), which is a linear polymer of beta(1,6)-linked N acetylglucosamine (GlcNAc) residues. Dispersin B (DspB), a soluble glycoside hydrolase produced by the periodontal pathogen Actinobacillus actinomycetemcomitans degrades PGA. The enzyme DspB is an alpha/beta TIM-barrel protein and belongs to family 20 glycosyl hydrolases members. The enzyme activity of DspB with regard to its substrate specificity towards beta(1,6)-linked GlcNAc polymers and its endo/exo character was investigated through ligand docking and the hydrolysis of synthetic oligosaccharides. Ligand docking analysis suggested that beta(1,6)-linked GlcNAc oligosaccharide bound to the active site better that beta(1,4)-linked GlcNAc oligosaccharide. Our combined results indicate that DspB is an exo-acting enzyme that hydrolyzes beta(1,6)-linked N-acetylglucosamine oligomers. PMID- 19133501 TI - Effects of hyperbaric oxygenation on cultured barley embryos. AB - Changes in relative water content (RWC), lipid peroxidation, proline and antioxidant system in relation to the tolerance to oxidative stress enzymes mediated high pressure were investigated in Hordeum vulgare L. cv. Tokak. For this purpose, mature embryos cultured on MS media were treated in a hyperbaric oxygenation chamber (approx. 59.06 feets, 2 kp/cm2) with pure oxygen for 60 minutes/day for a growth period of ten days in a plant growth chamber. Constitutive activities of SOD, APOX, GR and POX were higher in hyperbaric oxygenated (HBO) explants, being 96.07%, 28.57%, 77.77% and 54.14% for the 5th days; 95.78%, 40%, 37.5%, and 94.98% for the 10th days of culture, respectively, than in the control plants. Increase in SOD activity was also shown on polyacrilamide gel electrophoresis on the 10th day of application. Proline accumulation was increased in HBO-treated explants both on the 5th days (85.71%) and 10th days (37.14%) of treatment. MDA content was found to be higher in HBO treated explants both on the 5th (53.84%) and 10th (59.83%) days of culture. PMID- 19133502 TI - Trifluralin-induced disorganization of microtubular cytoskeleton alters the development of roots in Hordeum vulgare L. AB - The extensive use of herbicides in agriculture becomes an important factor in environmental pollution, especially in case of slowly degradable compounds. Some agents act on plants during a long period of time, even if a very low concentration of the herbicide remains in the soil. Here, we investigated the toxicological effect of a low concentration of dinitroaniline herbicide, trifluralin, on growing seedlings of Hordeum vulgare L. Trifluralin in concentration of 1 microg/ml inhibited root growth. The mitotic activity of meristematic cells was suppressed due to the retardation of metaphase progression -alteration that can be caused by cytoskeleton disorder. Using antibodies to alpha-tubulin, we investigated the distribution of microtubules in root meristem cells. During all stages of mitosis, the highly regular system of microtubular cytoskeleton observed in control cells was slightly disorganized. An examination of root structure using light and electron microscopy demonstrated that the cell walls did not form normally during cell division that led to the appearance of large multinucleated cells. Also, the premature (pathological) cell differentiation was induced by trifluralin. A part of differentiating cells showed intracellular structural changes that are consistent with programmed cell death. It seems that the development of alterations in trifluralin-treated roots was due to the microtubular cytoskeleton disorganization. PMID- 19133503 TI - The effect of silicon on the symptoms of manganese toxicity in maize plants. AB - The effect of exogenously applied silicon (Si) on plant growth, lipid peroxidation, total phenolic compounds and non-protein thiols was studied in two maize varieties (Zea mays L. vars. Kneja 605, 434) differing in sensitivity to excess manganese (Mn). Based on the density of brown spots per leaf area and relative shoot weight (RSW) used to define Mn tolerance var. Kneja 434 was found to be more Mn-tolerant than Kneja 605. The lipid peroxidation level and total phenolic compounds were enhanced with increasing Mn concentration in the nutrient solution. In addition, the Mn-sensitive var. Kneja 605 with markedly expressed first visible Mn toxicity symptoms had higher levels of total phenolic acids than var. Kneja 434 thus supporting the hypothesis that a stimulating effect of Mn on phenol content reflected rather a stress response to Mn excess than a tolerance mechanism. In contrast, non-protein SH content increased to a higher extent in the Mn-tolerant var. Kneja 434. The increased amount of non-protein SH compounds was accompanied by a much stronger oxidative stress in the Mn-sensitive plants when compared with the Mn-tolerant variety, thus suggesting that non-protein SH compounds may play a role in Mn tolerance in maize. The addition of silicon (Si) reduced the density of brown spots per leaf area as well as lipid peroxidation level and improved plant growth in Mn-treated plants. PMID- 19133504 TI - A novel poly(3-hydroxybutyrate)-degrading Streptoverticillium kashmirense AF1 isolated from soil and purification of PHB-depolymerase. AB - A new bacterial strain, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from soil. This organism, identified as Streptoverticillium kashmirense AF1, secreted PHB depolymerases both on solid as well as in liquid mineral salt medium containing poly(3-hydroxybutyrate) as sole carbon source. The optimum production of PHB depolymerase was observed at pH 8 and 7, at 45 degrees C, 1% substrate concentration and in the presence of lactose as an additional carbon source. The extracellular PHB depolymerase was purified by gel permeation chromatography using Sephadex G-75. The Streptoverticillium kashmirense AF1 produced two types of PHB depolymerases having molecular weights of about 37 and 45 kDa as determined by SDS-PAGE. The difference in dry cell mass and amount of CO2 evolved in the test and control calculated gravimetrically through Sturm test indicated the degradative capabilities of Streptoverticillium kashmirense AF1. PMID- 19133505 TI - Changes in the leaf polypeptide patterns of wheat flag leaves spraying with non purine cytokinins. AB - The changes in the polypeptide patterns of soluble proteins of wheat flag leaves after treatment with two synthetic cytokinin-like substances 3-CP-2-PU [1-(3 chlorophenyl)-3-(2-pyridyl)urea] and 3-CP-4-MPU [1-(3-chlorophenyl)-3-(4-methyl-2 pyridyl)urea] was studied. Both compounds increased differentially in quantity and composition most individual polypeptides identified compared to the non treated leaves. While 3-CP-2-PU increased only the polypeptide quantity, 3-CP-4 MPU led to the appearance of new polypeptide with Mw of 51 kDa. PMID- 19133506 TI - Gender differences in infertility-related stress and the relationship between stress and social support in Thai infertile couples. AB - OBJECTIVE: To study infertility-related stress among men and women and to examine its relationship with the level of perceived social support. MATERIAL AND METHOD: The Fertility Problem Inventory (FPI) and the Personal Resource Questionnaire (PRQ) were translated into Thai and used to assess the level of infertility related stress and perceived social support, respectively, in 238 infertile subjects. RESULTS: The global FPI scores for men and women were 154.2 +/- 18.3 and 154.7 +/- 22.6, respectively (p > 0.05). There was no significant difference in their perceived social support (PRQ scores = 137.8 + 14.0 and 134.0 +/- 16. 7 respectively). A significant negative correlation (r = -0.1894; p < 0.001) existed between global stress and social support in women, but not in men. CONCLUSION: Thai infertile couples experienced a high level of stress. Unlike previous studies from Western countries, there was no gender diference in infertility-related stress. PMID- 19133507 TI - Nomograms of the fetal heart between 16 and 39 weeks of gestation. AB - OBJECTIVE: To establish the normal fetal cardiac measurement from 16 to 39 weeks' gestation in Thai fetuses. MATERIAL AND METHOD: Four hundred and eighty normal singleton pregnant women were recruited into the present study. The sonographic measurements were obtained from axial scans at the level of the four-chamber view. RESULTS: The regression model for cardiac circumference (CC), thoracic circumference (TC), thickness of left ventricular wall during diastole (LVW), diameter of left ventricle during diastole (LVD) and systole (LVS) and thickness of interventricular septum during diastole (IVS) were best fit by allowing a polynomial for gestational age. The cardiothoracic ratio (CTR) and shortening fraction of left ventricle (SFLV) were not statistically different with advancing gestation. CONCLUSION: The normal values of fetal CC, TC, CTR, LVW, LVD, LVS, IVS and SFLV during gestation were established These data could serve as a standard to investigate cardiac change in pregnancies with fetuses at risk. PMID- 19133508 TI - Prognostic significance of ER, PR, Ki67, c-erbB-2, and p53 in endometrial carcinoma. AB - OBJECTIVE: To better discern the prognostic significance of estrogen-progesterone (ER-PR) receptor proliferative index, tumor suppressor gene, and over expression of oncogene c-erbB-2 in correlation with survival time and recurrence of tumor. MATERIAL AND METHOD: Paraffin blocks from 65 cases of endometrial carcinoma diagnosed and treatment at Rajavithi Hospital, Bangkok, Thailand with a follow-up time of at least 60 months were immunohistochemical studiedfor ER and PR status, tumor proliferative index (Ki-67), tumor suppressor gene (p53), and overexpression of oncogene c-erbB-2. Survival analysis was performed with the Cox proportional hazards. RESULTS: The mean age of the patients was 54.94 years with a range of 24 to 80 years. The mean follow-up time was 50.35 months. Nine patients (13.8%) had recurrent tumors, 5 years after treatment. Ten patients (15.4%) died of the primary disease during the follow-up period. ER was positive in 50 cases (76.9%) and negative in 15 cases (23.1%). PR was positive in 47 cases (72.3%) and negative in 18 cases (27.7%). Both ER and PR showed significant correlation (p<0.01). Ki-67 showed 27 cases (41.5%) having >35% positive nuclear staining and 38 cases (58.5%) had < or =35% positive nuclear staining. p53 was positive in 31 cases (47.7%) and negative in 34 cases (52.3%). c-erbB-2 was positive in one case (1.5%), equivocal in six cases (9.2%), and negative in 58 cases (89.3%). CONCLUSION: Survival analysis showed that cases with low-stage, low-grade, no recurrent tumor, ER and PR positive, and Ki-67 < or =35% had good survival compared to cases with high-stage, high-grade, presence of recurrent tumor, ER-PR-negative, and Ki-67 > 35% (p<0.05). Cox regression analysis showed ER-PR status and Ki-67 were significant independent prognostic indicators for survival time. Ki-67 expression was also a significant independent prognostic indicator for recurrent tumor p53 and c-erbB-2 displayed no statistical significance related to survival time. PMID- 19133509 TI - Reagent strip testing for antenatal screening and first meaningful of asymptomatic bacteriuria in pregnant women. AB - OBJECTIVE: To evaluate the diagnostic performance of reagent strip test as a screening test for asymptomatic bacteriuria (ABU) in pregnant women. MATERIAL AND METHOD: Three hundred and sixty asymptomatic pregnant women who attended their first antenatal appointment at Rajavithi Hospital from August 1st to October 31st, 2005 were enrolled Those with symptoms of urinary tract infection within one month, those who had been prescribed antibiotics during the previous 7 days, and those with medical or obstetric complications, vaginal bleeding and a history of urinary tract diseases were excluded Urine specimens were collected by clean catched midstream urine technique for urine dipstick and culture. RESULTS: The prevalence of ABU was 10.0% The urine dipstick nitrite leukocyte esterase and combined test had a sensitivity of 16.7%, 75.0% and 16.7%, specificity of 99.1%, 67.9% and 99.4%, positive predictive value of 66.7%, 20.6% and 75.0%, negative predictive value of 91.5, 96.1% and 91.5%, accuracy of 90.8%, 68.6% and 91.1%, respectively. CONCLUSION: Reagent strip testing indicated a fair sensitivity for routine antenatal screening for asymptomatic bacteriuria in pregnant women PMID- 19133510 TI - The accuracy of intraoperative frozen sections in the diagnosis of ovarian tumors. AB - OBJECTIVE: To determine whether frozen section evaluation of ovarian tumors can be used to establish a histopathologic diagnosis and guide the surgeon to perform the appropriate surgical procedure. MATERIAL AND METHOD: Between January 2002 and December 2006, 376 ovarian specimens were submitted for frozen section examination and received for permanent section evaluation. The accuracy, sensitivity, specificity, and positive and negative predictive values of the frozen sections were studied. RESULTS: The overall accuracy was 87.8% while the inaccuracy was 12.2%. The sensitivity for malignant tumors was 79.6%, the sensitivity for borderline tumors was 61.8%, and the sensitivity for benign tumors was 98.2%. The specificity for the benign tumors was 88.6%, 93.8% for borderline tumors, and 97.1% for malignancy The positive predictive value was 92.2% for benign tumors, 63% for borderline tumors, and 91.1% for malignant tumors. CONCLUSION: The present study confirms that frozen section diagnosis is a reliable method for the surgical management of patients with ovarian masses. PMID- 19133511 TI - The accuracy of late antenatal screening cultures in predicting intrapartum group B streptococcal colonization. AB - OBJECTIVE: To determine the accuracy of late antenatal (35-37 weeks) screening cultures in predicting intrapartum group B streptococcal (GBS) colonization in Rajavithi Hospital (RH). MATERIAL AND METHOD: From the September 1st, 2006 to November 30th, 2006 at RH, 360 pregnant women who fulfilled the specified criteria were selected from antenatal clinic. Swabs were cultured from the lower vagina and anorectum for GBS using Todd-Hewitt broth with 15 mcg/ml nalidixic acid and 8 mcg/ml gentamicin. When they were admitted in the labor room (LR) for labor; the cultures were repeated in the same way. RESULTS: 302 out of 360 cases had been cultured in LR. The prevalence of GBS in pregnant women at 35-37 weeks and delivery were 13.05% and 13.58% respectively. Fifty-three out of 302 cases were colonized with GBS from at least ANC and/or LR. So the overall prevalence of GBS colonization in pregnant women in this study was 17.55% (53/302). The sensitivity, specificity, positive and negative predictive values of late antenatal GBS culture were 70.73%, 95.40%, 70.73% and 95.40% respectively. There was no significant difference between PPV and NPV in any interval between antenatal and intrapartum cultures. CONCLUSION: Late antenatal screening cultures were not sensitive in predicting intrapartum GBS colonization status. PMID- 19133512 TI - Alcohol septal ablation for hypertrophic obstructive cardiomyopathy, the first cases series in Thailand. AB - BACKGROUND: Alcohol septal ablation (ASA) is an emerging percutaneous technique to treat patients with hypertrophic obstructive cardiomyopathy (HOCM) and refractory to medical therapy. ASA in Thailand has never been reported. MATERIAL AND METHOD: The authors reviewed clinical and hemodynamic data of four patients with HOCM before and after ASA performed between November 2007 and May 2008 in Songklanagarind Hospital. RESULTS: ASA without myocardial contrast echocardiography was done successfully in all four patients with HOCM and refractory to medical therapy. The authors used the anatomical characteristics of vessel and pressure-guided technique to identify the optimal septal perforator artery. The averages of left ventricular outflow tract (LVOT) peak/mean pressure gradients (PPG/MPG) were 105/56 before and 32/18 mmHg immediately after ASA, consecutively. There were further falls in LVOT PPG and MPG to averages of 14 and 8.5 mmHg respectively at 6-12 week follow-up. The mean absolute alcohol volume was 2.5 +/- 0.41 ml. Transient complete atrioventricular block occurred in one patient. All patients reported substantial symptomatic improvement. CONCLUSION: The authors reported the first cases series of HOCM patients who underwent ASA in Thailand ASA without myocardial contrast echocardiography in carefully selected patient is feasible, effective, and safe. PMID- 19133513 TI - Cardiac troponin-T in pre-end stage kidney disease. AB - BACKGROUND: Cardiac troponin T level (cTnT) is commonly increased in end stage chronic kidney disease (CKD) in the absence of acute myocardial infarction. There are few data available on serum cTnT concentration in patients with pre-end stage CKD. OBJECTIVE: To evaluate the correlation of cTnT level and severity of kidney disease in patients with CKD stage 3 and 4 and to evaluate whether there is a relationship between left ventricular mass index and cTnT level. MATERIAL AND METHOD: Patients (103) with CKD stage 3-4 between 26 and 85 years of age (mean 60.0 +/- 11.9) entered the present study. Serum cTnT determined using a third generation electrochemiluminescent immunoassay on an Elecsys 2010 analyzer (Roche Diagnostics Ltd.). All patients underwent echocardiography. Left ventricular hypertrophy (LVH) was considered when LV mass index exceeded 125 g/m2. RESULTS: Overall, 28 patients (28.2%) had cTnT > 0.01 micro/L and two patients (1.8%) had cTnT > 0.1 microg/L cTnT concentration was commonly increased in more severe CKD (9 patients in stage 3 and 20 patients in stage 4). LVH was not associated with increased cTnT (p=0.105). CONCLUSION: The present study demonstrated that the elevated cTnT > 0.01 microg/L is relatively common in patients with CKD stage 3-4 who do not require dialysis treatment, however serum cTnT level above > 0.1 microg/L is uncommon in this population. Increased serum cTnT is associated with decreased renal clearance but not LVH. PMID- 19133514 TI - Prevalence of depression among a population aged over 45 years in Chiang Mai, Thailand. AB - OBJECTIVE: To determine the prevalence of depression in Thai people of 45 years and over. MATERIAL AND METHOD: The presented project was a cross sectional study on the prevalence of depression and cognitive impairment in Chiang Mai. Door-to door interview technique was assigned in condition with multistage probability random sampling to obtain subjects that represent a population of Chiang Mai. The research was conducted between October 2004 and September 2005. Data were collected on subject that were 45 years old and over. All subjects were selected from all districts in Chiang Mai. Thai Mini Mental State Examination (TMSE) and Thai Beck Depression Inventory (BDI) were used as the assessment tool If the subjects had a TMSE score less than 24 points, it was assumed as a cognitive impairment. RESULTS: One thousand four hundred ninety two people, 610 males and 882 females, were enrolled in the present study. Their mean age was 59.7 +/- 10.4 years (45-88 year). The prevalence of depression only was 29.2%, the prevalence of cognitive impairment only was 5.63% and the prevalence of cognitive impairment with depression was 3.96%. The prevalence of depression increased with age. CONCLUSION: The prevalence of depression in Thai people of 45 years and over was 29.2% and increased with age. PMID- 19133515 TI - Serum cardiac troponin-T as a prognostic marker in septic shock. AB - BACKGROUND: Cardiac troponins are markers for myocardial injury. Sepsis and septic shock can cause myocardial injury. The significance of elevated cardiac troponin T (cTnT) in septic shock was evaluated in the present study. MATERIAL AND METHOD: Serum levels of cTnT were measured in 40 patients with septic shock from Rajavithi Hospital during a nine-month period between December 2004 and August 2005. Patients with acute myocardial infarction were excluded by clinical presentation, electrocardiography, and measurement of creatine kinase. RESULTS: The levels of cardiac troponin T were elevated in 17 patients (42.5%) and not elevated in 23 patients (57.5%). There were 28 deaths (70%) and 12 patients (30%) survived and were discharged from the hospital. cTnT-positive patients had a significantly higher mortality rate (I 7 of 17 patients in the cTnT-positive group and 11 of 23 patients in the cTnT-negative group; p<0.001). CONCLUSION: Elevated cTnT levels may be a marker for severity of illness and higher mortality in patients with septic shock. PMID- 19133516 TI - Thai falls risk assessment test (Thai-FRAT) developed for community-dwelling Thai elderly. AB - OBJECTIVE: To develop falls risk assessment test that is appropriate for community-dwelling Thai elderly, and to verify this test with the second set of population. MATERIAL AND METHOD: A cross-sectional study was performed in 270 elderly living in Bansrang subdistrict, Ayuttaya province to identify a combination of variables that effectively predicted fall status in order to develop the Thai-FRAT The Thai-FRAT was validated with a second set of population whose cohort data had been collected during 1997-2002 in the study named "CERB project". One hundred fifty six elderly subjects were recruited in the analysis. RESULTS: The newly developed Thai-FRAT was composed of six factors including "History of falls", "Impaired body balance", "Female", "Specific medication use", "Impaired visual acuity" and "Thai style house". Possible score of the Thai-FRAT ranged from 0-11. The best cutoff score identified by the receiver operating curve analysis was 4. Sensitivity and specificity were 0.92 and 0.83 respectively. The Thai-FRAT could predict recurrent fall after two years among the elderly subjects who had had a history of fall during the past six months in the second set of population. Association between the Thai-FRAT score and mortality was also shown. CONCLUSION: The Thai-FRAT is the first fall risk assessment test developed for Thai community-dwelling elderly. It is a valid and reliable measure of fall risk. An effect of environment on falls among Thai elderly was clarified in the present study. PMID- 19133517 TI - Fatty acid composition in breast milk from 4 regions of Thailand. AB - BACKGROUND: DHA contents in breast milk varied upon maternal dietary intakes. OBJECTIVE: To study DHA contents in breast milk in Thai lactating women from four diferent regions of Thailand. MATERIAL AND METHOD: 20 mL of hind milk from 40 lactating women from Bangkok, Chantaburi, Tak, and Surin were collected and analyzed for fatty acids contents by gas chromatography Dietary intake of lactating women after delivery until the present study was assessed by a food frequency questionnaire. Then, the average DHA intake was estimated from the diets by using the reference data. RESULTS: DHA contents in breast milk of mothers from Surin were higher than those from other areas. There were no correlations between history of DHA intake and DHA contents in breast milk. CONCLUSION: DHA contents in breast milk vary from region to region of the country. Local dietary intake and genetics might explain this contrast. PMID- 19133518 TI - Normal anal position index in Thai newborns. AB - BACKGROUND: The quantitative measurement of the normal anal position by anal position index (API) has been reported from various institutes. The existing API data appear to vary among the ethnic differences. To date, the normal anal position in Thai children has never been reported. OBJECTIVE: Aim of the present study was to establish the normal values of the API in Thai newborns. MATERIAL AND METHOD: A cross-sectional study of 403 normal newborn born at Srinakharinwirot University Hospital between November 2003 and September 2004 was performed Demographic data and anal position index are reported separately for each gender split. Correlations between API and other anthropometric parameters were tested. RESULTS: There was significant difference of API between males (0.51 +/- 0.07; 95% CI 0.50 to 0.52) and females (0.38 +/- 0.08; 95% CI 0.37 to 0.39). API was not different between preterm and term newborn in each gender split. API had no significant correlation with gestational age (r=0.018, p-value = 0.71), birth weight (r=0.014, p-value = 0.79), birth length (r=0.02, p-value = 0.69) and head circumference (r=0.047, p-value = CONCLUSION: Analposition indexes in Thai newborn infants are 0.51 and 0.38 in male and female, respectively API had no correlation with gestational age and other anthropometric parameters such as birth weight, length, and head circumference. PMID- 19133519 TI - Multicentered study of model of difficult endotracheal intubation by incident reports from university and non-university hospitals. AB - OBJECTIVE: To compare the characteristics, causative factors, outcomes, prevention, and suggested preventive strategies of difficult intubation between university (U) and general community (non-U) hospitals. MATERIAL AND METHOD: One thousand nine hundred and ninety-six reports were reviewed from Thai anesthesia incident monitoring study (Thai AIMS) conducted in 51 hospitals nationwide between January and June 2007. Thirty-four cases ofDI were reported from U hospitals and 69 cases from non-U hospitals. The described details on each report on dfficult intubation (DI) in adults undergoing general anesthesia were thoroughly reviewed by three reviewers to give their consensus opinions on causative factors, outcomes, contributing preventive factors, and strategies for corrections. Descriptive statistics were used for data analysis. RESULTS: Patient factors were the most common cause of DI (88% in U and 87% in non-U hospitals). Fifty percent of U and 51% of non-U DI cases were consequences of human errors, which were preventable and mostly based on knowledge (88% vs. 71%) and rules of practice (23% vs. 51%). Substitution of an intubating anesthesiologist, reducing the size of endotracheal tubes, and stylet guided technique were the three commonly used methods after DI. MacCoy laryngoscope, fiber optic-aided intubation, laryngeal mask airway and Frova introducer were commonly used as substitutes for the standard laryngoscope. Inadequate experience was the major problem of U hospitals, which required additional training to gain more skill. The most common problem ofDI in non-U hospitals was inadequate preanesthetic evaluation. Therefore, they required practice guidelines and experienced assistants in difFicult situations. CONCLUSION: Half of DI cases were preventable. DI cases in Non-U hospitals were mostly caused by inadequate preanesthetic evaluation. This indicates the necessities of providing practice guidelines and experienced assistants. In U hospitals, in-training practice of intubation should be performed under supervision. More advanced substitution techniques were applicable in U hospitals. PMID- 19133520 TI - The Thai Anesthesia Incident Monitoring Study (Thai AIMS) of endobronchial intubation: an analysis of 1996 incident reports. AB - OBJECTIVE: To analyze the clinical course, outcomes, contributing factor, corrective and preventive strategies of accidental endobroncheal intubation (EBI) in the Thai Anesthesia Incident Monitoring Study (Thai AIMS). MATERIAL AND METHOD: This was a prospective descriptive multicenter study of anesthesia related adverse incidents from 51 hospitals across Thailand from January to June 2007. Possible accidental EBI data were extracted and analyzed using descriptive statistics by 3 reviewers. RESULTS: Thirty-two cases (1.6%) of EBI were reported from a total of l996 Thai AIMS incidents. EBI occurred more often in females (71.9%). Most of the incidents happened in the operating theater (93.8%) and the most common surgical specialties were general and gynecological surgery (20.6% each). Two cases had hypoxemia and 1 case required respiratory supported postoperatively. Most incidents (65.6%) were first recognized via monitoring equipment which was detected by pulse oximeter (71.4%) and airway pressure measurement (4.8%). Ninety six percent of cases were considered preventable. Anesthetic factors and system factors were found to involve in 62.5% and 11.8% of incidents respectively. The major contributing factors were inexperience of the performers (84.4%), lack of knowledge (40.6%), haste (21.9%) and communication failure (9.4%). The incident would be minimized by having prior experience of incident, high awareness and experienced assistants available. Three main strategies to prevent the incident included additional training, improvement supervision and established guideline practice. CONCLUSION: Accidental endobronchial intubation was reported as 1.6% of anesthetic adverse event in Thai AIMS. Majority of the incidents were contributed by anesthesia and system factors. High awareness, experience of performers and additional training would decrease the incidents and improve anesthetic outcome. PMID- 19133522 TI - In vitro antimicrobial activity of phramongkutklao hydroxyapatite antibiotic pellet. AB - OBJECTIVE: Assess the antimicrobial activity of Phramongkutklao antibiotic hydroxyapatite pellets in vitro. MATERIAL AND METHOD: The selected bacteria used in the present study were the standard and drug-resistant strains of Staphylococcus aureus and Pseudomonas aeruginosa, which are the common organisms causing infection in Orthopedics. Phramongkutklao antibiotic hydroxyapatite pellets developed by the Orthopedics Department, Phramongkutklao Hospital contained either Vancomycin, Fosfomycin, or Gentamycin. Each preparation was placed on an agar plate inoculated with each bacterium. The inhibition zones were monitored in 24 hours. Then the pellets were moved onto the new inoculated plate every day for 28 days. RESULTS: Compared with the control group, Phramongkutklao antibiotic hydroxyapatite pellets had good inhibitory effect against S.aureus. Vancomycin hydroxyapatite pellets were able to maintain their activity for 28 days whereas Gentamycin hydroxyapatite pellets was effective for only three days. Fosfomycin hydroxyapatite pellets could inhibit MSSA for 13 days and MRSA for 25 days. On the contrary, all three Phramongkutklao antibiotic hydroxyapatite pellets had very low efficacy against Paeruginosa. CONCLUSION: Phramongkutklao antibiotic hydroxyapatite pellets were able to inhibit the growth of S. aureus both MSSA and MRSA while the activity against P. aeruginosa needs to be developed. PMID- 19133521 TI - Local perianal nerve block versus spinal block for closed hemorrhoidectomy: a ramdomized controlled trial. AB - OBJECTIVE: To compare analgesic effectiveness, postoperative pain, complications, and patients' satisfaction between two randomly allocated groups--one group that had local perianal nerve block and another group that had spinal block following closed hemorrhoidectomy. MATERIAL AND METHOD: Sixty-seven patients underwent elective hemorrhoidectomy. Of these, 33 were randomly allocated to receive spinal anesthesia (SA) while 34 received perianal local analgesia (LA) with bupivacaine. Pain measurement at 6 and 24 hours following hemorrhoidectomy, the quantity of postoperative analgesic medication administered, patient's satisfaction and complications were recorded. RESULTS: Among the patients who had SA, there were 5 patients (15.2%) who developed hypotension during surgery. There was no reported case of hypotension among those who had LA. There was no significant difference in degree of median postoperative pain at 6 hours (LA: 38 vs. SA: 50 with VAS; p = 0.09) and at 24 hours (LA: 31 vs. SA: 35 with VAS; p = 0.35) between the two groups. Patients had a high satisfaction on both anesthetic methods. Patients in the SA group required more parenteral analgesics (p = 0.03) and had a higher incidence of urinary retention than those in the LA group (SA: 30.3% vs. LA: 8.8%, p = 0.03). CONCLUSION: Local perianal nerve block for hemorrhoidectomy is feasible and safe and superior to spinal block due to a lower incidence of post op urinary retention and less requirement of parenteral analgesics post-op. PMID- 19133523 TI - Prevalence and management of cancer pain in Srinagarind Hospital, Khon Kaen, Thailand. AB - OBJECTIVE: Cancer pain remains an invisible problem in cancer care and our study aimed to document its prevalence, characteristics, and patterns of management at a tertiary care teaching hospital. STUDY DESIGN: Descriptive, prospective, cohort study. MATERIAL AND METHOD: We recruited 335 consecutive adult patients diagnosed with cancers, admitted to Srinagarind Hospital, between February and April 2004. All of the participants were interviewed, and their pain evaluated by direct assessment using a numeric rating scale. RESULTS: The overall prevalence of cancer pain prior to admission was 56.5%, and within the first 24 hours of admission 41.5%. Three-quarters (74%) of patients with pain reported improvement; however one-third of those with pain never received any pain control intervention. Moreover; about half of those with persistent pain only received treatment by requesting it and then only received simple analgesics. CONCLUSION: Cancer pain remains under-detected and under-treated in many patients. Pain monitoring on a regular basis as well as a training program on pain management should be considered as first-line tools for improving pain control among cancer patients. PMID- 19133524 TI - Dimensional guide to harvesting the radius for orofacial reconstruction. AB - OBJECTIVE: To provide an accurate harvesting guide to maintain the maximum possible safe dimensions of the radius for orofacial reconstruction using the osteocutaneous radial forearm free flap. MATERIAL AND METHOD: Two hundred radii of 100 adult formalin-embalmed cadavers (52 males, 48 females) were measured. RESULTS: The mean minimum circumference of the radius measured between the pronator teres and brachioradialis insertions was 41.0 +/- 4.3 mm. At this point, the mean medio-lateral diameter was 13.4 +/- 1.6 mm. The mean of the maximum harvested length of the radius between the pronator teres and brachioradialis insertions was 81.3 +/- 10.4 mm. On the average, the maximum harvested length of the radius was 34.9% of its total length. The minimum medio-lateral diameter of the radius was consistently 1-5 mm less than 40% of its minimum circumference. There was no significant dimensional difference between sides but gender difference existed in all measurements (p < 0.001), except the maximum harvested length of the radius. CONCLUSION: The present study may be helpful to guide reconstructive surgeons for safer harvesting of the radius using the osteocutaneous radial forearm free flap to repair orofacial defects. PMID- 19133525 TI - Post stroke shoulder subluxation and shoulder pain: a cohort multicenter study. AB - OBJECTIVE: To study the occurrence of shoulder subluxation, shoulder pain in stroke patients and identify factors associated to these conditions during rehabilitation period. MATERIAL AND METHOD: Stroke patients from 9 rehabilitation centers from March to December 2006 were enrolled in the present study. All subjects were registered for demographic data including risk factors and type of stroke. They were assessed for motor recovery, cognitive ability, functional ability, psychological reaction and quality of life by using Brunnstrom stage, Thai Mental State Examination (TMSE), Barthel ADL Index (BI), Hospital Anxiety and Depression Scale (HADS) and WHOQOL-BREF questionnaires respectively at the beginning and the end of the present study. The occurence of shoulder subluxation and shoulder pain were recorded and then were analyzed for the associated factors. All subjects received the conventional rehabilitation program until they reached their rehabilitation goals or discharge criteria. RESULTS: Of 376 stroke patients, 327 met the inclusion criteria, 62 patients (19%) were found to have shoulder pain and 122 (37%) patients had shoulder subluxation. Shoulder pain was significantly more frequent in subjects with shoulder subluxation (odds ratio (OR) 2.48, 95% confidence interval (CI) 1.38-4.46) and at 2-6 months after stroke onset (OR 4.0, 95% CI 2.06-7.79). Shoulder subluxation was significantly associated with hemorrhagic type of stroke (OR 2.06, 95% CI 1.08-3.93), loss of proprioceptive sensation (OR 3.03, 95% CI 1.26-7.29) and negatively associated with Brunnstrom's stage of arm recovery (OR 0.44, 95% CI 0.34-0.56). No significant functional and quality of life impact was found from these conditions. CONCLUSION: Post stroke shoulder pain and subluxation were common during the rehabilitation period. Shoulder pain significantly occurred within 6 months after stroke onset and increased risk in patients with shoulder subluxation. Shoulder subluxation was correlated with Brunnstrom's stage, proprioceptive loss and hemorrhagic type of stroke. PMID- 19133526 TI - Clinical characteristics of ADHD in Thai children. AB - OBJECTIVE: To study the clinical characteristics of children who were diagnosed as ADHD. MATERIAL AND METHOD: A retrospective chart review was conducted on 202 children who came to a child mental health clinic and were diagnosed as ADHD. RESULTS: Most cases were in the 6-12 years age group and came from small families with 1-2 children. Males outnumbered females (M:F = 3.4:1). One-fifth of the sample received previous psychiatric evaluation from other health professionals but parents needed 'second opinion'. The most frequent chief complaints were academic/learning problems. Almost one-fourth of the samples came for problems not directly related to ADHD. In this group the most frequent complaints were aggressive and oppositional behavior Comorbidity was found in 53.5%. More than half of the cases who took intelligence tests had an IQ below 90. Behavioral management was the only treatment modality in 38% of the sample. In 62% stimulants were instituted either at the beginning of treatment or as an "add-on" after behavioral management proved to be insufficient. Among cases that received stimulants, 28% needed the combination of other psychotropic medications, mostly antidepressant and anxiolytic drugs. CONCLUSION: A study of the clinical characteristics of ADHD in Thai children revealed male preponderance and high rates of non-ADHD presentations and comorbid conditions. Awareness of varied presentations of ADHD and proper treatment of comorbid conditions is imperative in the comprehensive care of ADHD children. PMID- 19133527 TI - A case report of tumor-induced osteomalacia: eight year followed-up. AB - Tumor-induced osteomalacia (TIO) is a rare but potentially curable disease. It is caused by excessive renal clearance ofphosphate induced by a substance secreted from the tumor Here, the authors report a Thai patient who presented with multiple pathologic fractures, low serum phosphorus, and low tubular maximum reabsorption of phosphorus/glomerular filtration rate (TmPO4/GFR). The clinical, biochemical and bone abnormalities improved 6 months after the surgery. Two years follow-up showed no recurrence of the disease. Physicians should be aware of this condition when encountering with adult onset osteomalacia. PMID- 19133528 TI - Bilateral acute onset myopia and angle closure glaucoma after oral topiramate: a case report. AB - OBJECTIVE: Describe bilateral acute onset myopia and angle-closure glaucoma as ocular adverse effects of topiramate. CASE REPORT: A 23 year-old woman developed bilateral severe blurred vision seven days after initiating therapy with topiramate. Her visual acuity was counting fingers in both eyes. Intraocular pressures were 33 mmHg and 32 mmHg in the right and left eyes, respectively, with conjunctival chemosis, corneal edema, shallow anterior chambers, and closed angles. Her refraction was -7.50 diopters in both eyes. The symptoms and clinical findings resolved completely upon discontinuation of topiramate and, administration of antiglaucoma drugs. CONCLUSION: Topiramate use can result in acute bilateral angle-closure glaucoma and myopia, which are usually reversible upon cessation of the drug. Visual outcome is usually good and the episode resolves within a few weeks. Thus, it is important for clinicians to recognize these conditions and educate patients about these serious adverse effects when prescribing topiramate. PMID- 19133529 TI - Optic neuritis in a patient with Miller-Fisher syndrome. AB - Miller-Fisher syndrome (MFS) is considered a variant of Guillain-Barrd syndrome (GBS). The syndrome is characterized by acute onset of gait ataxia, ophthalmoplegia, and areflexia. Conventionally, MFS has been considered exclusively a disease of the peripheral nervous system. However there are occasional reports of central nervous system involvement. Here, a 62 year-old woman who presented with classical features of MFS and progressive bilateral dyschromatopsia and reported visual impairment. Normal MRI of the brain and CSF albumino-cytologic dissociation were observed. CSF oligoclonal IgG pattern indicated a passive transfer of oligoclonal IgG from a systemic inflammatory response. Nerve conduction studies showed slow motor conduction velocity in her extremities. The optic fundi were normal. Visual evoked potentials (VEPs) revealed bilateral optic neuropathy. Marked spontaneous improvement of her syndrome was documented within six weeks. Optic neuritis may be a central nervous system feature that should be recognized as part of the MFS. PMID- 19133530 TI - Utility of plasma fluorometric emission scanning for diagnosis of the first 2 cases reports of variegate porphyria: a very rare type of porphyrias in Thai. AB - Two Thai women who are siblings presented with a history of recurrent pruritic vesicles on dorsum of both hands and extensor surface of forearms where the sun exposed areas are. The excoriated vesicles were healed with depressed scars. They had no previous history of intense abdominal pain, seizure, or psychiatric disorder Urinary porphyrins were analyzed by High Performance Liquid Chromatography (HPLC). The level of coproporphyrin III was detected to be higher than the uroporphyrin level. Fluorescence emission scanning of both patients' plasma was performed and demonstrated typical emission peak at 626 nm, that confirmed the diagnosis of variegate porphyria. PMID- 19133531 TI - Attempted suicide by massive insulin injection: a case report and review of the literature. AB - The authors present a case of an 80-year-old man, non-diabetic, who attempted suicide by injecting himself subcutaneously with 10,000 units of Humulin R and 6000 units of Humulin N. Administration of dextrose intravenously was required for 13 days to maintain the capillary blood glucose within the range of 100-180 mg/dl. Hyponatremia, hypokalemia, hypophosphatemia, and elevated liver enzymes were also seen after massive insulin injection. Glucose requirement index was established to demonstrate the trend of glucose requirement during hospitalization. He recovered completely without any complication after monitoring blood glucose and titrating intravenous glucose carefully for two weeks. Current literature about how to manage insulin overdose was reviewed in the present article. PMID- 19133532 TI - Guidelines for antiretroviral therapy in HIV-1 infected adults and adolescents: the recommendations of the Thai AIDS Society (TAS) 2008. AB - BACKGROUND: More than 100,000 patients have been treated, since the implementation of the National Universal Coverage for antiretroviral therapy (ART) in Thailand Although there are several comprehensive guidelines available internationally, there is a need to have guidelines that can be implemented in Thailand. MATERIAL AND METHOD: The guidelines were developed by a panel of 17 members who are the experts on HIV research and/or HIV patient care and appointed without incentive by the Thai AIDS Society (TAS). The recommendations were based on evidences from the published studies and availability of antiretroviral agents. Published studies that are relevant and applicable to Thailand in particular have been taken into consideration. RESULTS: The recommendations include: when to start ART; what to start; how to monitor the therapy; adverse effects and its management; diagnosis of treatment failure; and antiretroviral treatment options in patients with treatment failure. ART in special circumstances, i.e., patients with co-infection of tuberculosis or hepatitis B virus, is also included Appropriate level of CD4+ T-cell count to start ART among Thai patients has been considered carefully. The authors recommend to start ART at CD4+ T-cell count < 200 cells/mm3. CONCLUSION: ART should be initiated in adults and adolescents HIV-1 infected patients with a history of HIV-related illness or AIDS or with a CD4+ T-cell count <200 cells/mm3. For treatment-naive patients, the preferred initial therapy is a non-nucleoside reverse transcriptase inhibitor (NNRTI)-based regimen. CD4' T-cell count and viral load should be monitored for at least twice and once a year, respectively. Proper management of antiretroviral-related toxicity and enhancement of adherence are crucial for the long-term success of ART. PMID- 19133533 TI - [Characteristics of disturbances of intercortical and cortical-subcortical integration in various clinical forms of neurotic depression]. PMID- 19133535 TI - [Echo thresholds measured in the vertical and horizontal planes]. PMID- 19133534 TI - [Neurophysiological indices of the CNS plasticity in the course of treatment of craniocerebral trauma in adolescents]. PMID- 19133536 TI - [Parameters of cerebral blood flow of 7- to 11-year-old children living in Northern European Russia]. PMID- 19133537 TI - [Electrical activity of the brain and oxygen supply for cognitive-mnestic activity in humans under different degrees of hypoxia]. PMID- 19133538 TI - [Effects of olfactory stimuli on students with various tones of the autonomic nervous system]. PMID- 19133540 TI - [Psychophysiological characteristics of young workers at an aircraft plant]. PMID- 19133539 TI - [Chronophysiological study of occupational fatigue]. PMID- 19133541 TI - [External respiration function of young male residents of Northern Europe during the annual cycle]. PMID- 19133542 TI - [Microcirculation characteristics in apparently healthy subjects during acute hypoxia and intermittent hypoxic training]. PMID- 19133543 TI - [Electrode configuration influence on the velocity of the action potential transmission during registration of interference electromyogram]. PMID- 19133544 TI - [Effect of intense muscle exercise on motor responses to magnetic stimulation of the brain and the spinal cord]. PMID- 19133545 TI - [Various mechanisms of voluntary control of walking in humans]. PMID- 19133546 TI - [The compensatory role of increased air content of the lung respiratory regions]. PMID- 19133547 TI - [Effects of the production of perceived exertion during cycle ergometry]. PMID- 19133548 TI - [Physical work capacity of athletes with different types of adaptation of the cardiorespiratory system]. PMID- 19133549 TI - [Change opens perspectives]. PMID- 19133550 TI - [Nothing works anymore--or the addiction of gambling]. PMID- 19133551 TI - ["The clear hearing of nurses is often required"]. PMID- 19133552 TI - [Self-reliance of the nursing profession]. PMID- 19133553 TI - [Better care of the skin]. PMID- 19133554 TI - [Clarifying the role of psychiatric nurses]. PMID- 19133555 TI - [Unknown heroes preserve an image]. PMID- 19133557 TI - [Nurses represent "security measures"]. PMID- 19133556 TI - [Taking the measure of the supply of nursing]. PMID- 19133558 TI - ["Let me stir things up with fantasy and courage"]. PMID- 19133559 TI - [The essential record for intensive care patients]. PMID- 19133560 TI - [Record of the patient at home]. PMID- 19133561 TI - [A universal consideration is forced]. PMID- 19133562 TI - [New perspectives]. PMID- 19133563 TI - [Starting up: a difficult choice]. PMID- 19133564 TI - [Taking on responsibility]. PMID- 19133565 TI - Measurement of nasal nitric oxide in neonates. PMID- 19133566 TI - Intravenous pamidronate therapy in Taiwanese patients with osteogenesis imperfecta. AB - BACKGROUND: Information on the long-term efficacy of intravenous pamidronate therapy in Asian patients with osteogenesis imperfecta (OI) is limited. We report our experience using pamidronate in Taiwanese patients with OI. METHODS: Twenty six patients with type I, III, or IV OI (eight males and 18 females; age range at last follow-up, 2.9-39.2 years) who received (or were currently receiving) intravenous pamidronate treatment (30mg/m2/dose, every month) were retrospectively analyzed. Patients were followed for 1.0-7.3 years over the study period from February 2000 to October 2007. RESULTS: The mean standard deviation score (SDS) for bone mineral density (BMD) had increased significantly from -4.72 to -3.37 (p < 0.005) after 1 year of treatment. In 16 patients evaluated after 4 years and eight after 6 years, the mean SDS continued to improve, to -2.69 (p < 0.001) and -1.54 (p < 0.005), respectively. The fracture rate decreased significantly (from 2.8 +/- 1.1 to 0.6 +/- 0.6, p < 0.001), and nine patients (35%) had no fractures while receiving treatment. The response to pamidronate was significantly better in patients with poorer initial BMD SDS (1 year: r = -0.71, p < 0.01; 4 years: r = -0.81, p < 0.01). CONCLUSION: This retrospective study suggests that Taiwanese patients with OI can benefit from pamidronate treatment, leading to a reduced incidence of fractures and increased BMD, especially in patients with poor baseline BMD. PMID- 19133567 TI - Endogenous nitric oxide in the nasal airways of healthy term newborn infants in Taiwan. AB - BACKGROUND: Nitric oxide (NO) in the respiratory tract is an important regulator of pulmonary homeostasis during the perinatal transition. In humans, much of the nitric oxide is derived from the upper airways, and autoinhalation of nasal NO has been suggested to influence pulmonary function. No standard methods for measuring nasal NO in neonates currently exist, and previous studies have reported varying levels of nasal nitric oxide in infants, due to the different measuring methods used. The use of nasal olives with a central lumen placed securely in the nares was recommended as a standardized procedure for the measurement of nasal NO in adults and children in 1999. We attempted to establish a safe, convenient and practical method for measuring nasal NO in healthy newborn infants, and investigated possible correlations between nasal NO and gender, postnatal age, gestational age, birth weight, and the differences between the right and left nostrils. METHODS: Nasal NO was studied in healthy newborn infants within the first 3 postnatal days. Gas was sampled from the nostril, and NO concentrations were determined using a fast response chemiluminescence analyzer. Each newborn infant underwent NO measurements on the first, second and the third postnatal days. Ninety-one newborn infants completed the study. RESULTS: Peak nasal NO in 91 newborn infants was 42.52 +/- 16.82 (mean +/- SD) parts per billion (ppb) (right nostril) and 40.86 +/- 16.08 ppb (left nostril) on the first postnatal day, 48.75 +/- 17.64 ppb (right nostril) and 49.47 +/- 17.26 ppb (left nostril) on the second postnatal day, and 59.65 +/- 19.72 ppb (right nostril) and 59.29 +/- 20.09 ppb (left nostril) on the third postnatal day. Nasal NO increased significantly with postnatal age (p < 0.001). There were no significant differences in nasal nitric oxide between sexes, or in relation to gestational age or birth weight, or between left or right nostrils. CONCLUSION: We conclude that nasal NO increased significantly in the first 3 days of life. PMID- 19133568 TI - Effects of the immunomodulatory agent Cordyceps militaris on airway inflammation in a mouse asthma model. AB - BACKGROUND: Cordyceps militaris is a well-known fungus with immunomodulatory activity. It is generally used in traditional Chinese medicine to treat hemoptysis, bronchial or lung inflammation, and urogenital disorders. The purpose of our study was to evaluate the effect of cultivated C. militaris on airway inflammation in a mouse asthma model. METHODS: BALB/c mice were sensitized with intraperitoneal ovalbumin (OVA) on Days 0 and 14, and were then given intranasal OVA on Day 14 and Days 25-27. Randomized treatment groups of sensitized mice were administered C. militaris, prednisolone, montelukast, or placebo by gavage from Days 15-27. Airway hyperreactivity to aerosolized methacholine was determined. Bronchoalveolar lavage fluid and serum were analyzed to assess airway inflammation. RESULTS: OVA-sensitized mice developed a significant airway inflammatory response that was inhibited by prednisolone and montelukast, whilst C. militaris reduced airway inflammation less effectively. Airway hyperresponsiveness to methacholine was observed in OVA-sensitized mice and was reversed by both prednisolone and montelukast. C. militaris initially reversed airway hyperreactivity, but this effect disappeared at higher methacholine doses. CONCLUSION: C. militaris can modulate airway inflammation in asthma, but it is less effective than prednisolone or montelukast. These results demonstrate that C. militaris is unable to adequately block the potent mediators of asthmatic airway inflammation. PMID- 19133569 TI - Dislodgment of port-a-cath catheters in children. AB - BACKGROUND: Port-a-cath catheters are frequently used in children with malignancies. Their dislodgment is rare, but carries potentially serious risks. This study analyzed our 11-year experience of this important issue. METHODS: Between June 1997 and January 2008, 290 ports of different brands were implanted in children by pediatric surgeons. Among the patients, 12 children with catheter dislodgement were retrospectively studied. Their ages ranged from 2-16 years, with a median of 6.4 years. Their body weights ranged from 12-39 kg, with a median of 20 kg. Ten patients presented with a port-a-cath dysfunction, while the other two patients were identified incidentally during surgery for removal of their ports. RESULTS: The downstream ends of dislodged catheters were located in the right ventricle (five patients), right atrium (four), main pulmonary artery (one), left pulmonary artery (one) and right pulmonary artery (one). Eleven catheters were broken, and one catheter was disconnected from the port. Most (10/11) catheters were broken at the site of anastomosis to the port. All dislodged catheters were successfully retrieved without complications by transcatheter retrieval using a gooseneck snare. CONCLUSION: The dislodgment rate of port-a-cath catheters in children in our series was 4.1%. Most (83%) catheters were broken at the site of anastomosis to the port. All dislodged catheters could be successfully retrieved by transcatheter retrieval using a gooseneck snare. PMID- 19133570 TI - Applications of flexible bronchoscopy in infants with congenital vocal cord paralysis: a 12-year experience. AB - BACKGROUND: Congenital vocal cord paralysis (VCP) is a common cause of congenital stridor. Before the widespread application of flexible bronchoscopy (FB) by pediatricians, congenital stridor in infants was usually attributed to laryngomalacia. Prompt recognition and careful follow-up is crucial for the management of congenital VCP. METHODS: We performed a retrospective chart review of newborn infants with congenital VCP diagnosed by FB over a 12-year period. RESULTS: During the 12-year period, FB was performed on a total of 356 infants. Fifteen (4%) infants were diagnosed with congenital VCP. There were eight males and seven females and the mean age at diagnosis was 76.6 days. Stridor with respiratory distress was the most prominent presenting symptom. The majority (93%, 14/15) demonstrated bilateral VCP, while one patient (7%) had unilateral VCP. Seven of the 15 (46%) patients had idiopathic VCP, while eight (54%) had VCP associated with neuromuscular disorders. Tracheotomy was necessary in four patients (26%). None of them underwent further surgical interventions. Spontaneous recovery occurred in 10 patients (71%), and of these, 90% (9/10) were treated without tracheotomy. CONCLUSION: In order to allow prompt diagnosis of congenital VCP, FB should be performed in every newborn infant with stridor. Patients with congenital VCP should undergo additional imaging studies to detect any associated neurological abnormalities and intrathoracic comorbidities. The majority of patients can be managed conservatively and monitored carefully using serial FB. Corrective surgery should be reserved for those with a lack of resolution at prolonged follow-up, and those with significant comorbidities. PMID- 19133571 TI - Sacral dysgenesis associated with terminal deletion of chromosome 7 (q36-qter). AB - We report on the clinical, cytogenetic, and imaging findings in a patient with a 7q terminal deletion. The 11-year-old girl had mental retardation, microcephaly, a distinctive face, relatively small hands and feet, and sacral dysgenesis. High resolution GTG banding (550-850 bands) showed a 7q terminal deletion. A detailed evaluation of associated malformations and the overall clinical picture should be taken into account when identifying the underlying diagnosis in cases of sacral dysgenesis with mental retardation. PMID- 19133572 TI - Acquired duodenal obstruction in children. AB - Traumatic intramural hematoma of the duodenum is a rare cause of acquired duodenal obstruction in children, and a high degree of suspicion is therefore required to make an early and accurate diagnosis. We report a 6-year-old boy whose epigastrium was impacted by the handlebar of his bicycle during a traffic accident. The boy then experienced epigastralgia. Six days later, progressive bilious vomiting suggestive of gastrointestinal obstruction was noted. Imaging studies revealed a large hematoma extending from the fourth portion of the duodenum to the jejunum. Conservative methods of treatment failed to manage his condition. He underwent laparoscopic surgery to evacuate the hematoma. We also report a case of duodenal obstruction in a previously healthy 2-year-old girl who presented for the first time with acute symptoms of proximal intestinal obstruction. Contrast examinations showed apparent barium retention over the stomach and proximal duodenum. She underwent surgery due to persistent obstruction, and a mushroom-like foreign body was detected embedded in the orifice of the windsock duodenal web. After duodenoduodenostomy and removal of the bezoar, she had a smooth recovery and tolerated feeding well. We conclude that blunt abdominal trauma and incomplete duodenal obstruction, such as that caused by duodenal web, should be considered as possible causes of acquired proximal gastrointestinal obstruction in previously healthy children, despite their rarity. PMID- 19133573 TI - Spontaneous multiseptated cystic pneumomediastinum in a term newborn. AB - Pneumomediastinum usually occurs after assisted ventilation. Spontaneous pneumomediastinum without preceding assisted ventilation, and presenting as a huge cystic mass in a term newborn infant has rarely been reported. We report a term baby who developed respiratory distress at 2 days of age. A huge cystic tumor of the thymus or a congenital mediastinal cystic tumor was initially suggested by chest radiography and computed tomography. Follow-up chest film revealed a "spinnaker sail sign" which is a typical radiographic presentation of pneumomediastinum. PMID- 19133574 TI - Valproate-induced hyperammonemic encephalopathy. AB - Valproate-induced hyperammonemic encephalopathy is an unusual but serious complication that can occur in people with normal liver-associated enzyme levels, and despite normal therapeutic doses and serum levels of valproate. Here, we describe an adolescent girl suffering from absence seizures, who complained of progressive dizziness and general malaise several days after restarting valproate. She developed vomiting and decreased consciousness after 3 weeks of valproate use. She had a serum ammonia level five times higher than the upper normal limit, normal liver-associated enzymes, and a supra-therapeutic valproate level. Electroencephalography (EEG) showed continuous generalized slowing. Tandem mass spectrometry analysis revealed carnitine deficiency. Her consciousness improved after emergent hemodialysis. Her ammonia level and EEG also became normal. Possible mechanisms, risk factors and treatments of valproate-induced hyperammonemic encephalopathy are described. Physicians should consider this possibility when consciousness disturbance occurs in patients treated with valproate. PMID- 19133575 TI - [Snuff doesn't save lives - only smoking cessation does!]. PMID- 19133576 TI - [We must have clear apologies]. PMID- 19133577 TI - [Fewer check ups and re-appointments when Vasterbotten redistributes 114 millions]. PMID- 19133578 TI - [Risk with strict glucose control and hydroxyethyl starch in severe sepsis. Routine intensive care treatments are now questioned]. PMID- 19133579 TI - [Medical students admitted by interviews as good as the rest of the students in examination following internship. They were also younger at the time of the final examination]. PMID- 19133580 TI - [Snuff and blood pressure. Cross-sectional study of blood pressure in rest among men in the MONICA study in Northern Sweden]. PMID- 19133581 TI - [Arabinitol ratio in urine for early diagnosis of Candidiasis infection]. PMID- 19133582 TI - [High-dose statin in ischemic stroke reduces the risk of recurrent stroke]. PMID- 19133583 TI - [Burns]. PMID- 19133584 TI - [Drug interaction or adverse effect of Lapacho-te?]. PMID- 19133585 TI - [How do we quality-assure point-of-care diagnosis?]. PMID- 19133586 TI - [HPV test for quality control of gynecologic cytological tests]. PMID- 19133587 TI - [No value for money with certain antihypertensive agents]. PMID- 19133588 TI - [KI's decision on psychotherapy education from a Holy Graal to a holy dispute]. PMID- 19133590 TI - [Create a Committee of Inquiry for safety management in health care]. PMID- 19133589 TI - [Does LMV consider comparative clinical trials between generic and original preparations unnecessary?]. PMID- 19133591 TI - [Remarkable HSAN decision on rhinosinusitis]. PMID- 19133592 TI - [Stress symptoms and hyperreactivity--diagnosis and treatment of "culture diseases"]. PMID- 19133593 TI - [Tocilizumab in rheumatoid arthritis. Interleukin 6 as a new target]. PMID- 19133594 TI - [Nano-particles--pharmaceutical "dwarves" with know-how]. AB - Self-cleaning surface coatings, tooth paste with repair effect, mini fuel cells and extremely small data memories, which contain the knowledge of whole libraries: After "micro" in the 1980ies and "electronic" in the 1990ies, "nano" is the technological keyword of this decade. The new nano-materials fascinate laymen and experts alike. Also in pharmacy the advance into dimensions unattainable so far, paved the way for the formulation of new pharmaceutical preparations. The nanotechnology offers innovative answers to previously unresolved galenic and/or biopharmaceutical questions and offers unexpected possibilities for drug targeting. PMID- 19133595 TI - [Supplementation with folic acid before and during pregnancy]. AB - Since 1995, supplementation of folic acid (400 microg/d) for prevention of neural tube defects is recommended for women during or before pregnancy by the German Society of Nutrition. Nevertheless, most women in childbearing age and also other people in the German population have no adequate supply with folic acid. Furthermore, genetic polymorphisms of the methylenetetrahydrofolate reductase gene can elevate the risk of congenital defects, particularly in children of mothers with low levels of folic acid. To allow a sufficient supply with folic acid for persons with reduced enzymatic activity of the methylenetetrahydrofolate reductase, a supplementation with a combination of folic acid and 5 methyltetrahydrofolate can be performed. PMID- 19133596 TI - [Specific immunotherapy in allergy treatment. Evidence for subcutaneous and sublingual administration]. PMID- 19133597 TI - How SII may affect your PPR. PMID- 19133598 TI - Modifications to the electronic Statement of Conditions for states requiring suspension of testing activities. PMID- 19133599 TI - Policy and requirement changes for ambulatory surgical centers using the Joint Commission deemed status option. PMID- 19133600 TI - CORRECTION: new accreditation decision rule for long-term care. PMID- 19133601 TI - Joint Commission resources launches the flu vaccination challenge. PMID- 19133602 TI - Neoadjuvant chemotherapy for ovarian cancer: clarifying the confusion. PMID- 19133603 TI - First-line therapeutic strategies in metastatic colorectal cancer. AB - The treatment of metastatic colorectal cancer (mCRC) has changed dramatically from the 1980s, when only fluorouracil (5-FU) was available for treatment and the median survival was at best 12 months, to a time when mCRC is considered more of a chronic disease in which the median survival is now reported in excess of 2 years. This review traces the evolution of treatment in this setting, including studies of single-agent vs combination treatment with 5-FU/leucovorin, irinotecan, oxaliplatin, and capecitabine, comparisons of simultaneous and sequential regimens, and the role of targeted agents such as bevacizumab, cetuximab, and panitumumab. PMID- 19133604 TI - Management of small-cell lung cancer: incremental changes but hope for the future. AB - Over 25,000 people are diagnosed with small-cell lung cancer (SCLC) in the United States annually. SCLC is a highly aggressive tumor with a propensity for early metastases and a high case-fatality rate. Systemic treatment with etoposide plus a platinum agent is recommended for all stages of this disease and has been a standard first-line therapy for SCLC since the 1980s. Three recently presented randomized clinical trials failed to show superiority of newer regimens over etoposide and cisplatin. Patients with limited-stage (LS) disease benefit from the addition of radiotherapy to systemic chemotherapy, a combination that affords high complete response rates and potential cures. Incremental improvements in radiotherapy delivery over the past decade include the use of accelerated hyperfractionated thoracic radiotherapy for LS disease. Prophylactic cranial irradiation, previously recommended for patients with LS disease, has recently been shown to benefit those with extensive-stage (ES) disease as well. Surgery, largely abandoned in the 1970s, is being reevaluated as primary local therapy in patients with very early-stage SCLC. Topotecan remains the only US Food and Drug Administration-approved therapy for recurrent disease. Amrubicin has demonstrated single-agent activity in multiple phase II trials in both chemotherapy-sensitive and -refractory relapse. The past 2 decades have been marked by an improved understanding of SCLC biology, and these discoveries are reflected in the number and diversity of novel therapies entering early-phase testing in this disease. PMID- 19133606 TI - Management of a patient with borderline resectable pancreatic cancer. Case report and review. AB - The definition and management of borderline resectability for periampullary pancreatic adenocarcinoma are evolving. In this case report, we discuss the presentation, workup, and therapeutic management of a 40-year-old man who presented with borderline resectable, periampullary pancreatic cancer and underwent a margin-negative resection following neoadjuvant chemoradiotherapy. PMID- 19133607 TI - Complementary therapies, herbs, and other OTC agents. PMID- 19133605 TI - Adult Burkitt lymphoma: advances in diagnosis and treatment. AB - Burkitt lymphoma is a unique B-cell malignancy with a high proliferation rate and characteristic genetic changes involving the c-myc oncogene. Burkitt lymphoma is common in children but also occurs in adults, where distinction from diffuse large B-cell lymphoma may pose a problem. The development of brief, very intensive chemotherapy regimens has led to a very high cure rate in children with Burkitt lymphoma. The use of these regimens in adults, often in combination with the antibody rituximab (Rituxan), has also made the cure of a majority of adults possible. Burkitt lymphoma in adults cannot be treated effectively with the common regimens used for diffuse large B-cell lymphoma such as CHOP-R (cyclophosphamide, doxorubicin HCl, vincristine [Oncovin], prednisone, rituximab). Prompt diagnosis and initiation of appropriate therapy with attention to the possibility of tumor lysis syndrome are necessary for optimal results. PMID- 19133608 TI - Isoflavone regulates lipid metabolism via expression of related genes in OVX rats fed on a high-fat diet. AB - OBJECTIVE: To investigate the effects of isoflavone on body weight, fat mass, and gene expression in relation to lipid metabolism. METHODS: Thirty-six female SD rats were ovariectomized or sham-operated and fed on a high-fat diet. Two months later, abdominal incision was made, blood was collected to separate serum, and the liver and adipose tissue were immediately collected and weighed. Some portions of these tissues were frozen in liquid nitrogen and stored at -80 degrees C. RESULTS: Ovariectomy (OVX) with a high-fat diet could induce obesity in rats, while treatment with isoflavone significantly inhibited the increase in body weight and fat mass in abdomen. Serum total cholesterol and leptin were significantly decreased in isoflavone group, compared with the OVX group. The mRNA expression of liver fatty acid synthase (FAS) in the OVX group was significantly higher than that in sham-operated group, while this difference was not observed in the isoflavone group. The mRNA expression of liver hormone sensitive lipase (HSL) in the OVX rats tended to be lower than that in the sham operated rats. Furthermore, a large amount of isoflavone maintained the mRNA expression at a sham level. CONCLUSION: Isoflavone may prevent obesity induced by ovariectomy with a high-fat diet, in part by modulating gene expression related to lipid metabolism. PMID- 19133610 TI - Recombinant human IgG antibodies against human cytomegalovirus. AB - OBJECTIVE: To study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection. METHODS: Fab monoclonal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of V(H) and V(L) and neutralizing activity on HCMV AD169 stain were analyzed in vitro. The light and heavy chain Fd fragment genes of Fab antibodies were further cloned into a recombinant baculovirus expression vector pAC-kappa-Fc to express intact IgG. Secreted products were purified with affinity chromatography using protein G. RESULTS: SDS-PAGE and Western blot confirmed the expression of the intact IgG. Immuno-blotting and -precipitation were used to identify HCMV proteins. One Fab monoclonal antibody recognized a conformational HCMV protein. CONCLUSION: IgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 microg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans. PMID- 19133609 TI - Antagonistic potential against pathogenic microorganisms and hydrogen peroxide production of indigenous lactobacilli isolated from vagina of Chinese pregnant women. AB - OBJECTIVE: To investigate the indigenous lactobacilli from the vagina of pregnant women and to screen the isolates with antagonistic potential against pathogenic microorganisms. METHODS: The strains were isolated from pregnant women's vagina and identified using the API50CH system. The ability of the isolates to produce hydrogen peroxide was analyzed semi-quantitatively using the TMB-HRP-MRS agar. The antagonistic effects of the isolates on pathogenic microorganisms were determined with a double layer agar plate. RESULTS: One hundred and three lactobacilli strains were isolated from 60 samples of vaginal secretion from healthy pregnant women. Among them, 78 strains could produce hydrogen peroxide, in which 68%, 80%, 80%, and 88% had antagonistic effects against Candida albicans CMCC98001, Staphylococcus aureus CMCC26003, Escherichia coli CMCC44113, and Pseudomonas aeruginosa CMCC10110, respectively. CONCLUSION: The recovery of hydrogen peroxide-producing lactobacilli decreases with the increasing pregnant age and time. The most commonly isolated species from vagina of Chinese pregnant women are Lactobacillus acidophilus and Lactobacillus crispatus. Most of L. acidophilus and L. crispatus produce a high H2O2 level. PMID- 19133611 TI - Bioassay of estrogenic activity of effluent and influent in a farm wastewater treatment plant using an in vitro recombinant assay with yeast cells. AB - OBJECTIVE: Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on industrial discharges, while the impact of livestock wastes as a source of endocrine disrupters in aquatic environments has been rarely elucidated. In order to investigate the contribution of environmental estrogens from livestock, the estrogenic activity in water samples from a farm wastewater treatment plant was analyzed by a recombinant yeast screening method. METHODS: The extracts prepared from 15 selected water samples from the farm wastewater treatment plant, among which 6 samples were from pre-treatment section (influents) and 9 from post-treatment section (effluents), were analyzed for estrogenic activity by cellar bioassay. Yeast cells transfected with the expression plasmid of human estrogen receptor and the Lac Z reporter plasmid encoding beta-galactossidase, were used to measure the estrogen-like compounds in the farm wastewater treatment plant. RESULTS: The wastewater samples from influents showed a higher estrogenic potency than the effluent samples showing a low induction of beta-galactossidase relative to solvent control condition. By comparison with a standard curve for 17 beta-estradiol (E2), estrogenic potency in water samples from the influents was calculated as E2-equivalent and ranged from 0.1 to 150 pM E2-equivalent. The estrogenic potency in water samples from the effluents was significantly lower than that in the influents, and 7 water samples had less detectable limit in the total of 9 samples. CONCLUSION: Yeast bioassay of estrogenic activity in most of the samples from the farm wastewater after disposal by traditional sewage treatment showed negative results. PMID- 19133612 TI - Non-fusion and fusion expression of beta-galactosidase from Lactobacillus bulgaricus in Lactococcus lactis. AB - OBJECTIVE: To construct four recombinant Lactococcus lactis strains exhibiting high beta-galactosidase activity in fusion or non-fusion ways, and to study the influence factors for their protein expression and secretion. METHODS: The gene fragments encoding beta-galactosidase from two strains of Lactobacillus bulgaricus, wch9901 isolated from yogurt and 1.1480 purchased from the Chinese Academy of Sciences, were amplified and inserted into lactococcal expression vector pMG36e. For fusion expression, the open reading frame of the beta galactosidase gene was amplified, while for non-fusion expression, the open reading frame of the beta-galactosidase gene was amplified with its native Shine Dalgarno sequence upstream. The start codon of the beta-galactosidase gene partially overlapped with the stop codon of vector origin open reading frame. Then, the recombinant plasmids were transformed into Escherichia coli DH5 alpha and Lactococcus lactis subsp. lactis MG1363 and confirmed by determining beta galactosidase activities. RESULTS: The non-fusion expression plasmids showed a significantly higher beta-galactosidase activity in transformed strains than the fusion expression plasmids. The highest enzyme activity was observed in Lactococcus lactis transformed with the non-fusion expression plasmids which were inserted into the beta-galactosidase gene from Lactobacillus bulgaricus wch9901. The beta-galactosidase activity was 2.75 times as high as that of the native counterpart. In addition, beta-galactosidase expressed by recombinant plasmids in Lactococcus lactis could be secreted into the culture medium. The highest secretion rate (27.1%) was observed when the culture medium contained 20 g/L of lactose. CONCLUSION: Different properties of the native bacteria may have some effects on the protein expression of recombinant plasmids. Non-fusion expression shows a higher enzyme activity in host bacteria. There may be a host-related weak secretion signal peptide gene within the structure gene of Lb. bulgaricus beta galactosidase, and its translation product may introduce the enzyme secretion out of cells in special hosts. PMID- 19133613 TI - A rabbit model of hormone-induced early avascular necrosis of the femoral head. AB - OBJECTIVE: To establish an experimental model of early stage avascular necrosis of the femoral head (ANFH) caused by corticosteroid in adult rabbits and to observe the pathological changes with various imaging techniques. METHODS: ANFH was induced by a combination of hypersensitivity vasculitis caused by injection of horse serum and subsequent administration of a high dose of corticosteroid. The pathological changes were detected with digital radiography (DR), computed tomography (CT), magnetic resonance imaging (MRI), ink artery infusion angiography, hematoxylin-eosin staining, and immunohistochemistry. RESULTS: The imageological and pathological changes corresponded to the clinical characteristics of early stage ANFH. DR showed bilaterally increased bone density, an unclear epiphyseal line, and blurred texture of cancellous bone. CT showed spot-like low-density imaging of cancellous bone, thinner cortical bone, osteoporosis, and an unclear epiphyseal line. MRI showed bone marrow edema and spot-like high signals in T2-weighted imaging in cancellous bone. Ink artery infusion angiography showed fewer obstructed blood vessels in the femoral head. HE staining of pathological sections showed fewer trabeculae and thin bone, an increased proportion of empty osteocyte lacunae, decreased hematopoiesis, thrombosis, and fat cell hypertrophy. Immunohistochemistry showed attenuated expression of vascular endothelial growth factor in osteoblasts and chondrocytes, and on the inner membrane of blood vessels. CONCLUSION: Experimental rabbit model of early stage ANFH caused by corticosteroid can be successfully established and provide the foundation for developing effective methods to treat early stage ANFH. PMID- 19133614 TI - Vulnerability to HIV infection and related health risk behaviors of the out of school adolescents migrated with their parents to Beijing. AB - OBJECTIVE: To explore the vulnerability to HIV infection and related risk behaviors of the out-of-school adolescents who migrated with their parents from rural areas to cities. METHODS: From September to October 2005, 260 out-of-school adolescents aged 14-20 years were interviewed through a questionnaire. RESULTS: Out-of-school adolescents lacked HIV/AIDS knowledge and related life skills, and their psychosocial competency was relatively low. The interviewed adolescents were open in their sexuality, showing a low rate of condom use. Their attitudes towards people living with HIV/AIDS and HIV/AIDS prevention competency were positively correlated with family well-being, harmonious relations between their parents, a stable job, knowledge, life skill, and psychosocial competency for emotion control and empathy. CONCLUSION: Out-of-school adolescents are highly vulnerable to HIV/AIDS infection and transmission. Practical and feasible health and life skill education and psychosocial competency are the keys to effective HIV/AIDS prevention among out-of-school adolescent migrants. PMID- 19133615 TI - Parental alcoholism, adverse childhood experiences, and later risk of personal alcohol abuse among Chinese medical students. AB - OBJECTIVE: To determine the status of adverse childhood experiences (ACEs) and the association of multiple ACEs with both parental alcoholism and later personal alcohol abuse among Chinese medical students with a view of improving adolescent health and reducing alcohol abuse among them. METHODS: In this cross-sectional study, 2073 Chinese medical students completed a survey on ten categories of ACEs in Anhui province of China. The association of parental alcoholism with ACEs and personal alcohol abuse was assessed by logistic regression analyses. RESULTS: The adjusted odds ratio (OR) for each category of ACEs in the subjects whose parents (either fathers or mothers or both) had alcohol abuse was 2 to 14 times higher than that in those with parental alcoholism (P<0.05). Subjects with bi-parental alcoholism had the highest likelihood of ACEs. Compared with the subjects without ACEs, therisk of personal alcohol abuse was increased by 2-4-folds in the subjects with ACEs, irrespective of parental alcoholism (P<0.05). The total number of ACEs (ACE score) had a graded relationship to 4 categories of personal alcohol abuse with or without parental alcoholism. The prevalence of personal alcohol abuse among the subjects with parental alcoholism was higher, which was independent of ACE scores. CONCLUSION: The prevalence of ACEs is generally serious in China. Efforts should be made to prevent and treat children with ACEs and subsequently to reduce alcohol abuse and later problems. PMID- 19133616 TI - Experimental study of cell migration and functional differentiation of transplanted neural stem cells co-labeled with superparamagnetic iron oxide and BrdU in an ischemic rat model. AB - OBJECTIVE: To explore the migration of transplanted neural stem cells co-labeled with superparamagnetic iron oxide (SPIO) and bromodeoxyuridine (Brdu) using the 4.7T MR system and to study the cell differentiation with immuno-histochemical method in ischemic rats. METHODS: Rat neural stem cells (NSCs) co-labelled with SPIO mediated by poly-L-lysine and bromodeoxyuridine (BrdU) were transplanted into the unaffected side of rat brain with middle cerebral artery occlusion (MCAO). At weeks 1, 2, 3, 4, 5, and 6 after MCAO, migration of the labelled cells was monitored by MRI. At week 6, the rats were killed and their brain tissue was cut according to the migration site of transplanted cells indicated by MRI and subjected to Prussian blue staining and immunohistochemical staining to observe the migration and differentiation of the transplanted NSCs. RESULTS: Three weeks after transplantation, the linear hypointensity area derived from the migration of labelled NSCs was observed by MRI in the corpus callosum adjacent to the injection site. Six weeks after the transplantation, the linear hypointensity area was moved toward the midline along the corpus callosum. MRI findings were confirmed by Prussian blue staining and immunohistochemical staining of the specimen at week 6 after the transplantation. Flourescence co-labelled immunohistochemical methods demonstrated that the transplanted NSCs could differentiate into astrocytes and neurons. CONCLUSION: MRI can monitor the migration of SPIO-labelled NSCs after transplantation in a dynamical and non invasive manner. NSCs transplanted into ischemic rats can differentiate into astrocytes and neurons during the process of migration. PMID- 19133617 TI - Detection and molecular characterization of enteroviruses in Korean surface water by using integrated cell culture multiplex RT-PCR. AB - OBJECTIVE: To identify waterborne enteric viruses in Korean surface water. METHODS: Integrated cell culture (ICC)-multiplex reverse transcription-polymerase chain reaction (RT-PCR) was simultaneously designed to detect coxsackieviruses (CV), polioviruses (PV), and reoviruses (RV). ICC-multiplex RT-PCR and phylogenetic analysis were conducted using 21 total culturable virus assay (TCVA) positive sample-inoculated cell cultures. RESULTS: CV and RV were detected in 9 samples each, and 3 samples were positive for both CV and RV. PV was not detected in any sample. Molecular phylogenetic analysis of the VP1 gene sequences revealed that CV types B2 and B4 predominated in Korean surface water, and the nucleotide sequences of CV type B2 were clustered with those of CVs isolated from China and Japan. The results suggested that the evolution of these viruses occurred in a region-specific manner. CONCLUSION: CV and RV are detectable in Korean surface water, with a predominance of CV type B2, and the evolution of CV type B2 occur in a region-specific manner. PMID- 19133618 TI - Spatiotemporal property analysis of birth defects in Wuxi, China. AB - OBJECTIVE: To describe the temporal trends and spatial patterns of birth defects occurring in Wuxi, a developed region of China. METHODS: Wavelet analysis was used to decompose the temporal trends of birth defect prevalence based on the birth defect rates over the past 16 years. Birth defect cases with detailed personal and family information were geo-coded and the relative risk in each village was calculated. General G statistic was used to test the spatial property with different scales. RESULTS: Wavelet analysis showed an increasing temporal trend of birth defects in this region. Clustering analysis revealed that changes continued in the spatial patterns with different scales. CONCLUSION: Wuxi is confronted with severe challenges to reduce birth defect prevalence. The risk factors are stable and show no change with spatial scale but an increasing temporal trend. Interventions should be focused on villages with a higher prevalence of birth defects. PMID- 19133619 TI - Visible light induced photodegradation and phototoxicity of phloxine B and uranine. AB - OBJECTIVE: To determine the visible light-induced photodegradation kinetics of two xanthene photosensitizers, phloxine B and uranine, in solution and on the surface of silica TLC plates, and to examine the phototoxicity of residues of degradation, which could provide valuable safety data on the two photosensitizers and other xanthene chemicals when applied in the environment. METHODS: UV-Vis absorption during photodegradation was monitored with a Unico 2102 spectrophotometer. Organic content of samples was measured with a Shimadzu TOC 4100. Phototoxicity tests were carried out using Saccharomyces cerevisiae with the methods modified from Daniels. RESULTS: When phloxine B and uranine degraded in solution, their apparent rate constant k was 0.0019 and 0.0027 min(-1), respectively. The total organic carbon (TOC) content decreased by approximately 50% during the 8 h irradiation period, which led to a gradual decrease in phototoxicity of the residues. The photodegradation of photosensitizers on the surface of silica TLC plates was much faster than that in the solution. The apparent rate constant k and the half life of phloxine B were 0.0073 min(-1) and 95 min, respectively. CONCLUSION: Visible light can rapidly induce photodegradation of phloxine B and uranine. The phototoxicity of residues is also decreased. The environmental risk of applications of phloxine B and uranine is minimal. PMID- 19133620 TI - Effects of a regional Chinese diet and its vitamin supplementation on proliferation of human esophageal cancer cell lines. AB - OBJECTIVE: To study the effects of a local diet popular in Yanting region (YT diet) on the proliferation of two human cell lines (Eca-109 esophageal squamous cell carcinoma line and HL7702 normal liver epithelial cell line) in rats by a sero-physiological approach. METHODS: Male SD rats were divided into six groups and fed respectively with a conventional diet and the YT diet (one of the five experimental diets) supplemented with two vitamin mixtures (Mix. 1: vitamins A, E, and folic acid; Mix.2: mix.1 plus riboflavin and vitamin C) at two different doses. On the 30th day, sera were collected from the rats and added into a medium for cell culture, with 10% FBS used as a serum control. The effects were assessed by MTT assay, DNA synthesis and flow cytometry assays. RESULTS: Compared with the control, the sera from rats fed with the YT diet significantly promoted the proliferation of Eca-109 cells, which was, however, reversed by the supplementation with two vitamin mixtures at high doses. Surprisingly, the same treatment produced contrary effects on HL7702 cells as compared with Eca-109 cells. CONCLUSION: The sera from rats fed with the YT diet could promote the proliferation of human esophageal cancer cell line Eca-109, whereas the sera from those fed with the YT diet supplemented with vitamin mixtures might have inhibitory effects on the proliferation of Eca-109 cells. PMID- 19133621 TI - The role of nitric oxide in biocompatibility. AB - Inflammation is a major factor in the development of the foreign body response to biomaterials and medical devices, including compatibility with blood, and the mechanisms are obviously complex. It has been realised recently that nitric oxide (NO) gas is continuously generated by some cells in the body and that the molecules of this gas are able to modulate inflammatory and thrombotic processes. The challenge has then become one of incorporating NO releasing materials into medical devices to control biocompatibility. PMID- 19133622 TI - Trends in silicone tubing. AB - Silicone is the material of choice for many new applications. Those examined here include drug delivery and peristaltic pumps, pacemaker leads and tubing used in minimally invasive surgery and to deliver high pressure gases and liquids. PMID- 19133623 TI - Micro machining brain implant needle arrays. AB - A major challenge in the industry is the creation of a degree of control over electromechanical prostheses in severely disabled spinal injury patients or loss of-limb patients. Of the many techniques being researched to alleviate disabilities and damage of the nervous system, one approach is based on engineered electrodes interfacing with the brain. The case study described here illustrates the use of microelectrodischarge machining, which is a critical component in the creation of an engineering system that interlinks directly with the brain. PMID- 19133624 TI - The challenges of packaging combination devices. AB - This article focuses on the development of a packaging format for drug eluting stents where the package not only has to meet the needs of the stent, but also the needs of the drug incorporated into its polymer coating. The package has to allow the transfer of ethylene oxide gas for sterilisation, but when in storage, must provide a barrier to keep out moisture and oxygen. A pouch and commercial scale manufacturing process were developed to incorporate this dual function into one item. PMID- 19133625 TI - The evolving process of European combination product review, Part II. AB - A new European guideline on drug-eluting stents (DES) introduces for the first time detailed information that European drug authorities should review concerning the medicinal substance that is incorporated into this type of drug-device combination product. Part I of this article discussed European requirements that apply to DES. This article discusses the new guideline, other stent guidelines, and the evolving process of drug-device regulatory review in Europe. PMID- 19133626 TI - Outsourcing for product success and restful nights. AB - The potential benefits of outsourcing are well known, but the decision to contract with subcontract partner can be daunting. This article examines how to determine if a product is suitable for outsourcing and the steps to take to identify and qualify the correct outsourcing company. PMID- 19133627 TI - Validation and Six Sigma. AB - Medical device design and manufacturing organizations, and their suppliers, that are pursuing operational excellence are urged to employ Six Sigma. It is a validation activity that addresses risk and by harnessing this methodology companies can accelerate and improve their risk management. This article explains how. PMID- 19133628 TI - Approaches to the physico-chemical and mechanical characterisation of functional coatings, Part II. AB - Part 1 of this article demonstrated that knowledge of coating science is important in the formulation and early development of a functionally active coating for a medical device/combination product. Part II focuses on the testing challenges and verification of the finished coating on the product. PMID- 19133629 TI - The Irish medical device diagnostics industry in 2008. AB - The industry's continued success is the result of sound strategy and its application. The next tranche of planning extends up until 2013 and its important elements are outlined here. PMID- 19133630 TI - Advantages of automation in production and assembly. AB - How to achieve better products, greater product quality and increased profitability. PMID- 19133631 TI - Leadership plays a vital role in the dental profession. PMID- 19133632 TI - Is your practice at risk for an employee-related lawsuit? PMID- 19133633 TI - Biofilm formation and caries incidence with removable partial dentures. PMID- 19133634 TI - Flapless implant surgery for replacement of posterior teeth. PMID- 19133635 TI - Treating the edentulous mandible: immediately loaded implants. PMID- 19133636 TI - Smile makeover utilizing direct composite resin veneers. AB - Creating a beautiful smile is more than restoring a single tooth back to its proper form. One must take into account the entire aesthetic zone, along with the mechanics of restoring the teeth to proper form and function. To make this effort even more challenging, the clinician is in full control and completely accountable for making the direct composite resin restorations from which the smile is created. Patients usually won't critique the aesthetics of a posterior direct resin, but once we move into the visible smile (along with the fact that most cosmetic procedures are patient desire- and want-driven) we must be able to deliver what the patient expects. Preplanning the case and avoiding the "prep and pray" approach to the smile-design process is the cornerstone of success. Utilizing tools for the creation of the restorations, such as a preoperative wax up and silicone putty matrix, help the clinician break the procedure down to individual restorations that when created in harmony with the pre-operative design or wax-up, will allow a final "smile design" to emerge with predictability without getting lost in the daunting task of creating the entire smile all at once. Proper use of ideal composite materials adds the final touch on creating realistic results that even the most discerning patients demand. Layering colors, utilizing differing opacities and translucencies within the restorative process, is a must. Having a "recipe" to follow simplifies the process and gives the clinician confidence that the final result will have that realistic look. All in all, the easiest way to handle a challenging case is to break it down into smaller and more manageable increments in order to ensure a predictable outcome. PMID- 19133637 TI - Long-term composite provisionalization: the conversion. PMID- 19133638 TI - An alternative to gutta-percha for root canal obturation. PMID- 19133639 TI - Laser biopsy, decontamination, and hemostasis: a case report. PMID- 19133640 TI - Functional-cosmetic dentistry: a full-mouth smile makeover, part 2. PMID- 19133641 TI - Identifying the aesthetic and functional determinants in the collapsed dentition. AB - This patient's treatment involved a complex diagnostic challenge, as well as a challenging clinical sequence due to the utilization of immediate implant placement and restoration after extraction along with immediate prosthodontic restoration with sinus elevation with bone grafting. The inability to have diagnostic patient wax try-ins required a detailed and exacting diagnostic work up which included significant laboratory diagnostic wax-ups. The utilization of the ACP Parameters of Care for Partial Edentulism for a PDI Class IV patient provided a framework in which care could be planned and executed with confidence. The availability of various reconstructive materials and techniques to create a seamless restorative result is essential to the success of this type of advanced treatment. PMID- 19133642 TI - You're the only chance I have! PMID- 19133643 TI - Flexible removable partial dentures: design and clasp concepts. PMID- 19133644 TI - Dental tourism: a growing concern. PMID- 19133645 TI - Radiologic-pathologic correlation of hepatocellular carcinoma treated with internal radiation using yttrium-90 microspheres. AB - We present the correlation between radiologic and pathologic findings in HCC patients who underwent radioembolization with yttrium-90 ((90)Y) microspheres prior to resection or transplantation. Thirty-five patients with a total of 38 lesions who underwent liver explantation after (90)Y radioembolization were studied. Imaging surrogates following treatment were evaluated; the explants were examined for assessment of necrosis by pathology. The correlation between radiologic and histologic findings of the treated lesions was analyzed. Twenty three of 38 (61%) target lesions showed complete pathologic necrosis. All target lesions demonstrated some degree of histologic necrosis at explant. Complete histologic necrosis was seen in 89% of lesions with pretreatment size <3 cm. Complete pathologic necrosis was seen in 100%, 78%, and 93% of the lesions that were shown to have complete response by European Association for the Study of the Liver (EASL) necrosis criteria, partial response by World Health Organizaton (WHO) criteria, or thin rim enhancement on posttreatment imaging, respectively. In contrast, complete pathologic necrosis was seen in only 52% and 38% of the lesions that showed partial response by EASL criteria and peripheral nodular enhancement, respectively. CONCLUSION: Post-radioembolization imaging findings of response by EASL and WHO criteria are predictive of the degree of pathologic necrosis. Rim enhancement was an imaging characteristic that correlated well with histologic necrosis. PMID- 19133646 TI - Computer-assisted image analysis of liver collagen: relationship to Ishak scoring and hepatic venous pressure gradient. AB - Histopathological scoring of disease stage uses descriptive categories without measuring the amount of fibrosis. Collagen, the major component of fibrous tissue, can be quantified by computer-assisted digital image analysis (DIA) using histological sections. We determined relationships between DIA, Ishak stage, and hepatic venous pressure gradient (HVPG) reflecting severity of fibrosis. One hundred fifteen patients with hepatitis C virus (HCV) who had undergone transplantation had 250 consecutive transjugular liver biopsies combined with HVPG (median length, 22 mm; median total portal tracts, 12), evaluated using the Ishak system and stained with Sirus red for DIA. Liver collagen was expressed as collagen proportionate area (CPA). Median CPA was 6% (0.2-45), correlating with Ishak stage (stage 6 range, 13%-45%), and with HVPG (r = 0.62; P < 0.001). Median CPA was 4.1% when HVPG was less than 6 mm Hg and 13.8% when HVPG was 6 mm Hg or more (P < 0.0001) and 6% when HVPG was less than 10 mm Hg and 17.3% when HVPG was 10 mm Hg or higher (P < 0.0001). Only CPA, not Ishak stage/grade, was independently associated by logistic regression, with HVPG of 6 mm Hg or more [odds ratio, 1.206; 95% confidence interval (CI), 1.094-1.331; P < 0.001], or HVPG of 10 mm Hg or more (odds ratio, 1.105; 95% CI, 1.026-1.191; P = 0.009). CPA increased by 50% (3.6%) compared with 20% in HVPG (1 mm Hg) in 38 patients with repeated biopsies. CONCLUSION: CPA assessed by DIA correlated with Ishak stage scores and HVPG measured contemporaneously. CPA was a better histological correlate with HVPG than Ishak stage, had a greater numerical change when HVPG was low, and resulted in further quantitation of fibrosis in cirrhosis. PMID- 19133647 TI - The quest for liver fibrosis biomarkers: promises from the enhanced liver fibrosis panel and beyond. PMID- 19133648 TI - Quantitative T2 mapping during follow-up after matrix-associated autologous chondrocyte transplantation (MACT): full-thickness and zonal evaluation to visualize the maturation of cartilage repair tissue. AB - The purpose of this article was to evaluate the potential of in vivo zonal T2 mapping as a noninvasive tool in the longitudinal visualization of cartilage repair tissue maturation after matrix-associated autologous chondrocyte transplantation (MACT). Fifteen patients were treated with MACT and evaluated cross-sectionally, with a baseline MRI at a follow-up of 19.7 +/- 12.1 months after cartilage transplantation surgery of the knee. In the same 15 patients, 12 months later (31.7 +/- 12.0 months after surgery), a longitudinal 1-year follow up MRI was obtained. MRI was performed on a 3 Tesla MR scanner; morphological evaluation was performed using a double-echo steady-state sequence; T2 maps were calculated from a multiecho, spin-echo sequence. Quantitative mean (full thickness) and zonal (deep and superficial) T2 values were calculated in the cartilage repair area and in control cartilage sites. A statistical analysis of variance was performed. Full-tickness T2 values showed no significant difference between sites of healthy cartilage and cartilage repair tissue (p < 0.05). Using zonal T2 evaluation, healthy cartilage showed a significant increase from the deep to superficial cartilage layers (p < 0.05). Cartilage repair tissue after MACT showed no significant zonal increase from deep to superficial cartilage areas during baseline MRI (p > 0.05); however, during the 1-year follow-up, a significant zonal stratification could be observed (p < 0.05). Morphological evaluation showed no significant difference between the baseline and the 1-year follow-up MRI. T2 mapping seems to be more sensitive in revealing changes in the repair tissue compared to morphological MRI. In vivo zonal T2 assessment may be sensitive enough to characterize the maturation of cartilage repair tissue. PMID- 19133649 TI - Ljungan virus present in intrauterine fetal death diagnosed by both immunohistochemistry and PCR. AB - OBJECTIVES: Following up on prior evidence from animal and human studies of Ljungan virus (LV) in intrauterine fetal death (IUFD), we examine additional cases of IUFD using two standard assays of viral detection: immunohistochemistry (IHC) and real time RT-PCR. MATERIALS AND METHODS: Frozen and formalin-fixed specimens from IUFD cases were tested for the presence of LV using real time RT PCR and IHC, respectively. Formalin-fixed organs from terminated pregnancies diagnosed as trisomy 21 were used as controls in the IHC assay. RESULTS: Presence of LV was demonstrated in all five IUFD cases by IHC and further confirmed in three of these cases by real time RT-PCR. Only one of 18 trisomy 21 controls was LV positive by IHC. CONCLUSION: The presence of LV in IUFD patients has been confirmed by two different assays. PMID- 19133650 TI - An early complement-dependent and TLR-4-independent phase in the pathogenesis of ethanol-induced liver injury in mice. AB - The innate immune system has been implicated in the pathogenesis of alcoholic liver disease. Although innate immunity is usually considered an early response to injury, previous work implicating innate immunity in ethanol-induced liver injury focuses primarily on long-term ethanol exposure. We investigated the early period of ethanol exposure to determine whether there were temporal associations between activation of innate immune responses and known correlates of liver injury. Female C57BL/6 mice were allowed free access to an ethanol-containing Lieber-DeCarli diet or were pair-fed a control diet. Within 4 days of ethanol exposure, we observed a striking spike in expression of hepatic proinflammatory cytokines-including tumor necrosis factor alpha (TNF-alpha), interleukin-6, and interferon-gamma-prior to hepatic triglyceride accumulation or increased plasma alanine aminotransferase activities, as well as before the induction of cytochrome P450 2E1 or oxidative stress. This early spike in inflammatory cytokines coincided with deposition of C3b-iC3b/C3c (C3b) in the liver. This deposition, resulting from the cleavage of the third component of the complement system (C3), is evidence for activation of complement in response to ethanol. C3( /-) mice were protected from the early, ethanol-induced increase in hepatic TNF alpha expression. Ethanol increased C3b deposition in mice deficient in C3a receptor or C5a receptor, as well as in wild-type mice depleted of hepatic macrophages; however, there was no increase in hepatic TNF-alpha in the absence of C3a receptor, C5a receptor, or hepatic macrophages. In contrast, the absence of Toll-like receptor 4 (TLR-4) had no effect on the early, ethanol-induced increase in either C3b or TNF-alpha. CONCLUSION: We have identified a complement- and macrophage-dependent, but TLR-4 independent, phase in the pathogenesis of ethanol-induced liver injury. PMID- 19133651 TI - MicroRNA-101 regulates expression of the v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) oncogene in human hepatocellular carcinoma. AB - MicroRNAs (miRNAs) have recently been proposed as a versatile class of molecules involved in regulation of various biological processes. Although there is emerging evidence that some microRNAs can function as oncogenes or tumor suppressors, the specific role of miRNA in human hepatocellular carcinoma (HCC) is unclear at this point. In this study, we examined the microRNA expression profiles in a set of 20 human HCC specimens by miRNA microarray and quantitative real-time polymerase chain reaction. The results showed that among the 20 HCC samples analyzed, microRNA-101 was significantly down-regulated twofold or more (twofold to 20-fold) in 16 samples compared with the matching nontumoral liver tissues. Using both a luciferase reporter assay and Western blot analysis, we showed that microRNA-101 repressed the expression of v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) oncogene, a key component of the activator protein-1 (AP-1) transcription factor. Moreover, using a luciferase expression vector (pAP-1-Luc) driven by seven copies of an AP-1 cis-element, we observed that microRNA-101 expression inhibited phorbol 12-myristate 13-acetate (PMA)-induced AP-1 activity. In in vitro Matrigel invasion and Transwell migration assays, enhanced microRNA-101 expression inhibited the invasion and migration of cultured HCC cells, respectively. These findings suggest that microRNA-101 may play an important role in HCC. CONCLUSION: MicroRNA-101, which is aberrantly expressed in HCC, could repress the expression of the FOS oncogene. PMID- 19133652 TI - Bone lesions: role of sediment cytology. AB - This study was conducted to evaluate the role of sediment cytology of biopsy specimen fixatives, which is usually discarded, in early diagnosis of bone lesions. Cytological smears prepared from sediments of biopsy specimen fixatives (sediment cytology) were used to study 65 bone specimens biopsied with suspicion of malignancy. The cytological diagnosis was then compared with histological diagnosis, taking the latter as gold standard. Smears were adequately cellular and showed good preservation of cellular morphology. Some of the smears showed microfragments of tissue. Cytology labeled 29 lesions as malignant, 26 lesions as benign, 3 as inflammatory, and 7 smears as inconclusive because of low cell yield. Sediment cytology was able to correctly diagnose 58 of 65 lesions. There was no false-positive or false-negative case. The sediment cytology could be considered as an easy and effective diagnostic tool that can provide early diagnosis for the lesion of bone. PMID- 19133653 TI - Responsiveness to levodopa in epsilon-sarcoglycan deletions. AB - Myoclonus-dystonia (M-D) is characterized by early-onset myoclonus and dystonia, and is often due to mutations in the epsilon-sarcoglycan gene (SCGE) at locus 7q21. The pathogenesis of M-D is poorly understood, and in a murine knockout model, dopaminergic hyperactivity has been postulated as a mechanism. We present two unrelated individuals with M-D due to SCGE deletions who displayed a robust and sustained response to levodopa (L-dopa) treatment. In contrast to using dopamine blocking agents suggested by the hyperdopaminergic knockout model, we propose that a trial of L-dopa may be considered in patients with myoclonus dystonia. PMID- 19133654 TI - Recurrent hemichorea following a single infarction in the contralateral subthalamic nucleus. PMID- 19133655 TI - Parkinsonism and impulse control disorder: presentation of a new progranulin gene mutation. PMID- 19133657 TI - Effects of inhibitory rTMS on bladder function in Parkinson's disease patients. AB - Patients affected by Parkinson's disease (PD) may present with lower urinary tract (LUT) dysfunction characterized by involuntary detrusor overactivity. We evaluated possible impact of a 2-week course of low frequency 1 Hz repetitive transcranial magnetic stimulation (rTMS) on LUT behavior in eight advanced PD patients complaining of urinary disturbances. We tested the effects of rTMS measuring urodynamic examination and the International Prostate Symptoms Score (IPSS) questionnaire, used for evaluation of subjective LUTS. rTMS was able to improve temporarily LUT behavior in PD patients, increasing bladder capacity and the first sensation of filling phase. Moreover, a reduction of IPSS score was noticed, due to an improvement on filling phase symptoms. The beneficial effects assessed with the IPSS lasted for up to 2 weeks after the end of the stimulation. rTMS seems to be an effective, noninvasive alternative treatment for PD patients with urinary disturbances. PMID- 19133656 TI - Cancer surveillance behaviors and psychosocial factors among long-term survivors of breast cancer. Cancer and Leukemia Group B 79804. AB - BACKGROUND: Little is known about cancer surveillance (mammography, clinical breast examination, and pelvic examination) behaviors in long-term (9-16 years) breast cancer survivors. This report describes the relation of these behaviors to demographic and clinical characteristics, psychological symptoms, body satisfaction, and social support. METHODS: Survivors who had participated in Cancer and Leukemia Group B treatment Trial 8541 completed a survey that included questions on breast cancer surveillance and pelvic examination, psychological well being, body satisfaction, and social support. RESULTS: The participation rate was 78% and included 245 breast cancer survivors. Survivors (n = 107; 44%) reported completing breast cancer surveillance (mammography and clinical breast examination) and completing pelvic examination (n = 162; 68%) within recommended guidelines. There were no significant associations between breast cancer surveillance and breast cancer anxiety, depression, stressful life events, body satisfaction, social support, or demographic characteristics. Survivors within recommended guidelines for pelvic examinations were younger (P = .05), married (P = .003), had health insurance (P = .004), and had lower depression scores (P = .005) than survivors who underused or overused pelvic examination. In addition, survivors within recommended pelvic examination guidelines had significantly lower levels of breast cancer anxiety (P = .03) compared with survivors who underused pelvic examination. CONCLUSIONS: Many long-term breast cancer survivors were not within recommended cancer surveillance guidelines. Private health insurance was associated with following recommendations for pelvic examinations, although such a relation did not exist for breast cancer surveillance. The results of this study have implications for the development of educational programs to improve cancer surveillance among the growing population of long-term breast cancer survivors. PMID- 19133658 TI - Examination of the Lille Apathy Rating Scale in Parkinson disease. AB - Apathy is a unique, multidimensional syndrome commonly encountered in patients with Parkinson disease (PD). Recently, the Lille Apathy Rating Scale (LARS), a semistructured interview yielding a global score, and composite subscores for different domains of apathy (i.e., cognitive, behavioral, affective, self awareness), was developed and given to a sample of patients with PD in France. This study is the first outside of its original developers to examine the English language version of the LARS in PD. We found the LARS to be a coherent instrument demonstrating both convergent and divergent validity, as compared to the Apathy Scale (AS) and Beck Depression Inventory (BDI-II). Using a receiver operating characteristic (ROC) analysis comparing the LARS to the AS, a validated and widely-used measure, we identified a cut-off score (sensitivity = 64%, specificity = 92%, PPV = 88%, NPV = 75%) that was higher than that proposed by the original authors, who derived their cut-off by comparing LARS global scores to clinical judgments of apathy. Although the present study does not compare the LARS to a diagnostic gold standard or promote its utility for diagnosing apathy, it provides further support for the LARS as a promising instrument to examine apathy in PD. PMID- 19133659 TI - FGF20 and Parkinson's disease: no evidence of association or pathogenicity via alpha-synuclein expression. AB - Genetic variation in fibroblast growth factor 20 (FGF20) has been associated with risk of Parkinson's disease (PD). Functional evidence suggested the T allele of one SNP, rs12720208 C/T, altered PD risk by increasing FGF20 and alpha-synuclein protein levels. Herein we report our association study of FGF20 and PD risk in four patient-control series (total: 1,262 patients and 1,881 controls), and measurements of FGF20 and alpha-synuclein protein levels in brain samples (nine patients). We found no evidence of association between FGF20 variability and PD risk, and no relationship between the rs12720208 genotype, FGF20 and alpha synuclein protein levels. PMID- 19133660 TI - Drug hoarding: a case of atypical dopamine dysregulation syndrome in a RLS patient. PMID- 19133661 TI - Clinically deployable Kinesia technology for automated tremor assessment. AB - The objective was to design, build, and assess Kinesia, a wireless system for automated assessment of Parkinson's disease (PD) tremor. The current standard in evaluating PD is the Unified Parkinson's Disease Rating Scale (UPDRS), a qualitative ranking system typically completed during an office visit. Kinesia integrates accelerometers and gyroscopes in a compact patient-worn unit to capture kinematic movement disorder features. Objectively quantifying PD manifestations with increased time resolution should aid in evaluating efficacy of treatment protocols and improve patient management. In this study, PD subjects performed the tremor subset of the UPDRS motor section while wearing Kinesia. Quantitative kinematic features were processed and highly correlated to clinician scores for rest tremor (r(2) = 0.89), postural tremor (r(2) = 0.90), and kinetic tremor (r(2) = 0.69). The quantitative features were used to develop a mathematical model that predicted tremor severity scores for new data with low errors. Finally, PD subjects indicated high clinical acceptance. PMID- 19133662 TI - Funding of Parkinson research from industry and US federal and foundation sources. AB - Funding for biomedical and neuroscience research has increased over the last decade but without a concomitant increase in new therapies. This study's objectives were to determine the level and principal sources of recent funding for Parkinson disease (PD) research and to determine the current state of PD drug development. We determined the level and principal sources of recent funding for PD research from the following sources: US federal agencies, large PD foundations based in the United States, and global industry. We assessed the status of PD drug development through the use of a proprietary drug pipeline database. Funding for PD research from the sources examined was approximately $1.1 billion in 2003 and $1.2 billion in 2005. Industry accounted for 77% of support from 2003 to 2005. The number of drugs in development for PD increased from 67 in 2003 to 97 in 2007. Of the companies with at least one compound in development for PD in 2007, most were small (62% had annual revenue of less than $100 million), and most (53%) were based outside the United States. These companies will likely require partnerships to drive successful development of new PD therapies. PMID- 19133663 TI - Traditional Chinese medicine on four patients with Huntington's disease. AB - Four Huntington's disease (HD) patients were treated with traditional Chinese medicines Yi-Gan San (YGS) and Chaihu-Jia-Longgu-Muli Tan (CLMT) in a cross-over manner. Two patients took YGS for 8 weeks first, and after 4 weeks of washing out, they took CLMT for 8 weeks. Two other patients took these medicines in reverse order. All patients showed a decrease in the Unified Huntington's Disease Rating Scale-motor assessment (from 106.3 +/- 4.7 to 89.6 +/- 5.8 as mean +/- SD, P = 0.0004) by YGS treatment with no extrapiramidal symptoms or changes of cognition or ADL. Our study suggests a possibility of a new treatment for involuntary movements. PMID- 19133664 TI - Reported mutations in GIGYF2 are not a common cause of Parkinson's disease. PMID- 19133665 TI - Psychotic and compulsive symptoms in Parkinson's disease. AB - The objective of this study is to evaluate psychiatric symptoms in Parkinson's disease (PD) patients and to assess their relation with other clinical aspects of PD. Psychotic symptoms (PS) and compulsive symptoms (CS) as well as other nonmotor and motor features were evaluated in 353 PD patients. Psychotic and compulsive symptom scores did not correlate significantly. PS occurred in 65% of patients, with item frequencies ranging from 10% (paranoid ideation) to 55% (altered dream phenomena). Regression analysis showed that autonomic impairment accounted for 20% of the 32% explained variance of PS, whereas cognitive problems, depression, daytime sleepiness, and dopamine agonist (DA) dose explained the rest. CS occurred in 19%, with item frequencies of 10% for both sexual preoccupation and compulsive shopping/gambling. Patients with more severe CS (score > or = 2 on one or both items) were significantly more often men, had a younger age at onset, a higher DA dose and experienced more motor fluctuations compared to the other patients. PS and CS are common but unrelated psychiatric symptoms in PD. The relations found between PS and cognitive problems, depression, daytime sleepiness, and autonomic impairment suggests a resemblance with Dementia with Lewy Bodies. The prominent association between PS and autonomic impairment may be explained by a shared underlying mechanism. Our results confirm previous reports on the profile of patients developing CS, and mechanisms underlying motor fluctuations may also play a role in the development of CS in PD. PMID- 19133666 TI - Orthostatic hypotension, balance and falls in Parkinson's disease. AB - Orthostatic hypotension (OH) is a common feature in Parkinson's disease (PD). As the control of balance and gait is already affected by PD per se, OH may further predispose patients to falls and accidents. The study was conducted to evaluate the clinical correlates of OH and its association with mobility and balance in PD. From a total population of 205,000 inhabitants, 120 PD patients were included in the study. Medical data including history of recent falls were collected, and patients were clinically examined using the orthostatic test, the Timed Up & Go test, walking speed, and the quantitative measurement of postural sway. Sixty three (52.5%) patients had OH in the orthostatic test. Twenty-five (39.5%) patients with and 16 (28.1%) patients without OH (P = 0.614) had fallen during the past 3 months. Patients with OH had significantly increased postural sway in standing compared with patients without OH. However, OH was not associated with mobility or walking speed. The current results support the concept that the control of body balance and OH may be closely linked. PMID- 19133667 TI - Transcatheter closure of patent foramen ovale with radiofrequency: acute and intermediate term results in 144 patients. AB - AIMS: Currently available devices for transcatheter closure of patent foramen ovale (PFO) which rely on a permanent implant have limitations, including late complications. The study objective was to evaluate the safety, feasibility, and effectiveness of the PFx Closure System, the first transcatheter technique for PFO closure without an implantable device. METHODS AND RESULTS: A prospective study of 144 patients was conducted at nine clinical sites from October 2005 through August 2007. All patients had a history of cryptogenic stroke, transient ischemic attack, migraines, or decompression illness. The mean balloon stretched diameter of the PFO was 7.9 +/- 2.5 mm. Technical success (successful application of radiofrequency energy) was achieved in 130 patients. One patient required a transfusion as a result of blood loss during the procedure. There were no other major procedural complications. There were no recurrent strokes, deaths, conduction abnormalities, or perforations following the procedure. At a mean follow-up of 6 months, successful closure was achieved in 79 patients (55%). In PFOs with balloon sized or stretched diameters less than 8 mm, the closure rate was 72% (53/74). CONCLUSION: This study demonstrates that transcatheter closure of a PFO without a permanent implant is technically feasible and safe. Further technique and device modifications are required to achieve higher closure rates. PMID- 19133668 TI - Evaluation of intermediate coronary stenosis with intravascular ultrasound and fractional flow reserve: Its use and abuse. AB - Clinical decision making in patients with intermediate coronary stenosis is still debated. Intravascular ultrasound (IVUS) examination and/or functional assessment of coronary stenosis by fractional flow reserve (FFR) are currently used to define the severity of such lesions. There are very few studies with a small sample size that have a head-to-head comparison between IVUS and FFR in the evaluation of angiographically de novo intermediate lesions. There are no randomized, controlled trials to demonstrate the superiority of IVUS versus FFR in providing improved clinical outcomes in comparison with angiography alone. However, the issue of superiority might be irrelevant, because IVUS and FFR could be complementary techniques to be used in the catheterization laboratory to provide critical anatomic and functional data that permit more accurate decisions in the management of the patient. PMID- 19133669 TI - Stenting for a symptomatic posterior cerebral artery stenosis. AB - Evolvement of endovascular devices and increase of operator expertise have made angioplasty and stenting in intracranial vessels technically possible. Stenting has been reported in treating stenosis in middle and anterior cerebral arteries with favorable outcomes. However, the feasibility of stenting for stenosis in posterior cerebral artery (PCA) has not been established. We report a patient with progressive focal cerebral ischemic symptoms, which were arrested after reconstruction of the associated PCA stenosis with stenting. PMID- 19133670 TI - Contrast administration in pediatric cardiac catheterization: dose and adverse events. AB - BACKGROUND: In pediatrics, contrast-related AE such as allergic reactions, seizures, and nephropathy have been reported to occur after cardiac catheterization, but their incidence remains unknown. OBJECTIVE: We sought to report adverse event (AE) rates attributed to contrast administration in a pediatric cardiac catheterization lab and identify characteristics related to higher doses. METHODS: A single institution prospective cardiac catheterization AE database identified AE in children <18 years old exposed to contrast. All AE were reviewed and classified by relationship to contrast. Medical records for the 50 cases who received highest contrast doses were retrospectively reviewed for AE. Patient and procedural characteristics were compared in the top quartile of contrast dose versus remaining cases. RESULTS: Over 3 years, 2,321 consecutive cases required median 3.9 cm(3)/kg [IQR: 2.0, 6.0] of contrast. Patients receiving high dose contrast (top quartile) were more likely to be <1 year (51% vs. 24%), weigh <10 kg (66% vs. 29%), have complex 2 ventricle disease (56% vs. 35%), be in a high procedure type risk group (57% vs. 26%), and undergo procedures >2 h (67% vs. 28%), all P < 0.001. Only 2 of 2,321 cases (0.09%, 95% CI 0.01-0.31%) had AE possibly related to contrast. These events were an acute neurological change and transient nephropathy. In 50 cases receiving the most contrast, no AE were attributed to contrast. CONCLUSION: A large volume pediatric cardiac catheterization lab administered >or= 6 cm(3)/kg of contrast in a quarter of cases; however, AE related to contrast exposure were exceedingly rare. PMID- 19133671 TI - Percutaneous closure of aortic pseudoaneurysm by Amplatzer occluder device-case series of six patients. AB - Thoracic pseudoaneurysms are rare variety of aortic disorders. Although mostly asymptomatic, they represent potentially fatal conditions (Sullivan et al., Chest 1988;93:138-143; Razzouk et al., Ann Surg 1993;59:818-823) that are traditionally treated surgically. False aneurysms of aorta are usually a late complication of previous surgical procedure (Sullivan et al., 1988)-especially reconstructive surgery, trauma (Razzouk et al., 1993), and rarely infection (Sanchez-Recalde et al., J Am Coll Cardiol 2003;41:152-154). Surgical management is often complicated by poor outcomes with high morbidity and mortality (Mulder et al., Arch Surg 1998;133:45-49). Endovascular treatment is emerging as promising options for aortic diseases with fewer complications. We report a series of six cases at a single center where Amplatzer device was used to treat thoracic aortic pseudo aneurysm. To our knowledge only one series of three cases (Kanani et al., Catheter Cardiovasc Interv 2007;69:146-153) and few case reports (Bashi et al., Catheter Cardiovasc Interv 2005;65:547-551) of successful closure have been published previously. Our case series is the largest, so far, including mortality and specifically the preprocedural and postprocedural CT angiographic assessment of the pseudoaneurysms. PMID- 19133672 TI - Novel self-expanding stent system for enhanced provisional bifurcation stenting: Examination by StentBoost and intravascular ultrasound. AB - A 62-year-old man underwent percutaneous coronary intervention of a bifurcation lesion (Medina type 010) involving the mid-left anterior descending coronary artery and an important first diagonal branch with a novel stent specifically designed for bifurcations, the Stentys coronary bifurcation system. This is a self-expanding nitinol stent, with Z-shaped struts linked by interconnections that can be disconnected (in prespecified points every 1.5 mm all around the circumference and the length of the stent) at the level of the ostium of the side branch, simply by inflating an angioplasty balloon tracked to the ostium of the side branch, through the stent struts. The steps required for deployment of the stent and the final result obtained were evaluated by intravascular ultrasound examination and StentBoost Subtract, a specific X-ray stent-enhancing visualization technique. PMID- 19133673 TI - Continuous epoprostenol therapy and septal defect closure in a patient with severe pulmonary hypertension. AB - A 31-year-old woman with exertional dyspnea diagnosed as having atrial septal defect (ASD) with severe pulmonary hypertension (PH). Intravenous epoprostenol therapy was started to improve PH. Although pulmonary arterial pressure decreased, her symptoms remained in class III of WHO functional class, probably because of exacerbation of the left-to-right shunt caused by the reduction of pulmonary vascular resistance (PVR). Transcatheter atrial septal closure was therefore performed. Soon after the procedure, additional reduction in pulmonary arterial pressure was achieved. Her symptoms improved and oxygen inhalation was discontinued. One year after the procedure, although intravenous epoprostenol was still required, her symptoms had improved to class I of WHO functional class without exacerbation of PH. Transcatheter atrial septal closure after lowering PVR by intravenous epoprostenol would be a novel therapy for patients with ASD accompanied by PH. PMID- 19133674 TI - Transcatheter closure of atrial septal defect in elderly patients with permanent atrial fibrillation. AB - OBJECTIVES: The aim of this study is to evaluate the feasibility and efficacy of device closure of atrial septal defect (ASD) in elderly patients with permanent atrial fibrillation. BACKGROUND: Little is known about the feasibility of device closure of ASD in those patients. METHODS: Nine consecutive patients (mean age 68.1 years) with permanent atrial fibrillation (>1 year persistent) underwent catheter closure using the Amplatzer septal occluder. Transthoracic echocardiography and plasma B-type natriuretic peptide (BNP) level were assessed before and at 24 hours; and 1, 3, and >6 months after the closure. Before the procedure, appropriate dose of warfarin was used in all, diuretics was used in 8/9. Same amount of medications were continued after the procedure. RESULTS: ASD could be closed in all (mean device size 27.3 mm) without hemodynamic and thromboembolic complications. New York Heart Association (NYHA) functional classification was significantly improved in all patients after device closure. No hemodynamic and thromboembolic complications were observed during the follow up period (mean 10.6 months). Although permanent atrial fibrillation did not change in all after the procedure, resting heart rate decreased from 76.2 +/- 16.0 to 68.3 +/- 13.2 beats/min (P = 0.015). There was statistically significant improvement in right ventricular/left ventricular diameter ratio (1.08 +/- 0.16 to 0.73 +/- 0.10, P = 0.008) and plasma BNP level (183.7 +/- 90.5 to 94.6 +/- 47.4 pg/mL, P = 0.008) after >6 months device closure. CONCLUSIONS: Even in the patients complicated with permanent fibrillation, transcatheter closure of ASD can contribute to symptomatic improvement as well as cardiac geometric remodeling. PMID- 19133675 TI - Multiple coronary artery fistulae treated with a single Amplatzer vascular plug: check the back door when the front is locked. AB - Congenital coronary artery fistulae are rare anomalies, which can result in myocardial ischemia or infarction, congestive heart failure, fistula rupture, or death. In this report, the authors describe a 56-year-old woman with new onset chest pain and palpitations. Exercise myocardial perfusion imaging was significant for a reversible perfusion defect in the anterior left ventricular wall. Left and right heart catheterization demonstrated multiple fistulous communications between the right coronary and left anterior descending coronary arteries with the pulmonary artery. All fistulae drained into the pulmonary artery by a common ampulla. Closure was achieved via a percutaneous approach using a single Amplatzer vascular plug. PMID- 19133676 TI - Is routine ultrasound guidance really necessary for closure of patent foramen ovale using the Amplatzer PFO occluder? AB - OBJECTIVES: The aim of the study was to evaluate safety, efficacy, and long-term clinical outcome of percutaneous closure of patent foramen ovale (PFO closure) in a low volume center using the Amplatzer PFO occluder without echocardiographic guidance. BACKGROUND: Most centers perform PFO closure either by transesophageal echocardiography (TEE) or intracardiac echocardiography (ICE) guidance for optimal device selection. As TEE is poorly tolerated by patients in supine position and ICE is a costly alternative that increases vascular access complications, we wanted to assess the safety and efficacy of PFO closure by fluoroscopic guidance only. METHODS: Before PFO closure, all patients had a diagnostic contrast-TEE and morphological classification of PFO. All PFO closures were performed using the 25-mm Amplatzer PFO occluder with fluoroscopic guidance only. Intraprocedural echocardiography was replaced by right atrial opacification using contrast angiography. Contrast TEE was done after 6 weeks, contrast TTE after 3, 6, and 12 months postprocedural. RESULTS: In all 92 patients (52.4 +/- 1.5 years), a 25-mm Amplatzer PFO occluder was implanted in the correct position. Total fluoroscopic time was 8.4 +/- 0.6 minutes and the application of contrast medium was 122.5 +/- 5.8 mL. By contrast-TEE, 12 patients (13 %) showed a small residual shunt (grade 1). During follow-up (2.09 +/- 0.13 years) two patients (2.1%) suffered from a recurrent event (TIA in both cases). CONCLUSIONS: Percutaneous closure of PFO using the 25-mm Amplatzer PFO occluder guided by fluoroscopy only is a safe and efficacious intervention for nearly all patients. PMID- 19133677 TI - Chronotropic impairment after surgical or percutaneous closure of atrial septal defect. AB - BACKGROUND: In previous studies, an attenuated heart rate response to exercise has been noted in patients after surgical closure of atrial septal defect. The aim of this study was to compare the prevalence of chronotropic impairment after surgical and percutaneous closure of atrial septal defect. METHODS: Thirty-eight pediatric patients who underwent a surgical (group A, n = 18) or transcatheter closure (group B, n = 20) of atrial septal defect in our institution were prospectively included in the study. Treadmill exercise testing was performed using the Bruce walking treadmill protocol to voluntary exhaustion, with continuous monitoring of heart rate and oxygen consumption. RESULTS: All the children were in sinus rhythm, and had normal values for peak oxygen uptake and endurance time. Exercise heart rate was significantly lower than normal in group A at the end of stage 2 (Z-score = -0.71 +/- SD 1.02), and in both groups at the end of stage 3 (Z-score = -2.06 +/- 1.76 in A and -1.00 +/- 0.71 in B) and at peak exercise (Z-score = -2.78 +/- 2.14 in A and -0.81 +/- 0.75 in B). However, the heart rate response to exercise was significantly less attenuated in group B than in group A. Moreover, maximal heart rate was <-2 SD in 8/18 surgical patients but in no patient of the group B. CONCLUSION: The chronotropic impairment is significantly less important after transcatheter closure of atrial septal defect than after surgical closure. It is an additional argument in favor of the interventional catheterization. PMID- 19133678 TI - A novel 0.010-inch guidewire for crossing jailed sidebranches. AB - Complete occlusion of a side branch sometimes occurs due to a jailed-stent during treatment of bifurcation lesions. In this report, we describe a novel 0.010-inch coronary guidewire (Decillion) that is effective for crossing a completely occluded side branch through the jailed-stent strut, whereas another standard 0.014-inch guidewire was unable to cross the same lesion. This novel 0.010-inch guidewire may be useful for crossing of completely occluded side branches that are difficult to cross through the stent strut after stent implantation. PMID- 19133680 TI - Concomitant renal and iliac fibromuscular dysplasia. AB - Fibromuscular dysplasia (FMD) usually involves the renal arteries and is an important cause of secondary hypertension in younger women. We report an unusual case of FMD in a middle-aged woman, not only involving both the renal and iliac arteries but also presenting in a pattern more typical of atherosclerosis. PMID- 19133679 TI - Characterization of a Dazl-GFP germ cell-specific reporter. AB - In this study, we characterized the promoter activity of a 1.7 kb sequence in the 5' flanking region of the mouse Deleted in Azoospermia-Like (Dazl) gene. We found the 1.7 kb sequence sufficient to drive robust germ cell-specific expression of green fluorescent protein (GFP) in adult mouse testis and lower levels of expression in adult ovary and in fetal and newborn gonads of both sexes. This expression pattern was confirmed in two independently-derived transgenic mouse lines. In adult testis, Dazl-GFP exhibited a developmentally-regulated, stage specific expression pattern during spermatogenesis. GFP was highly expressed in spermatocyte stages, with strongest expression in pachytene spermatocytes. Weaker expression was observed in round and elongating spermatids, as well as spermatogonial cells. In the fetal gonad, GFP transcript was detected by e12.5 in both sexes; however, GFP fluorescence was only detected during later embryonic stages. In addition, we produced mouse embryonic stem cell (ESC) lines harboring the Dazl-GFP reporter and used this reporter to isolate putative germ cell populations derived from mouse ESCs following embryoid body differentiation and fluorescence activated cell sorting. PMID- 19133681 TI - Sudden cardiac death complicating alcohol septal ablation: a case report and review of literature. AB - Over the years, alcohol septal ablation has become an effective and well-accepted modality in the treatment of patients with hypertrophic obstructive cardiomyopathy refractory to standard medical therapy. Malignant tachyarrythmias infrequently complicates the procedure and are usually self-terminating. We describe a case of alcohol septal ablation complicated by sudden cardiac death occurring immediately following the procedure requiring prolonged resuscitative efforts with eventual complete recovery. We also discuss the pathophysiologic significance of this event in the setting of this cardiomyopathy and its relevance as a complication of the procedure. PMID- 19133682 TI - Re: One year perspective on COURAGE. PMID- 19133683 TI - Use of magnetic guidewire navigation in the treatment of lower extremity peripheral vascular disease: report of the first human clinical experience. AB - The use of a magnetic navigation system (MNS) to facilitate catheter placement in electrophysiologic procedures has been increasingly documented over the past decade. Recently, the utilization of MNS to aid guidewire directionality in complex coronary lesions has also been described. Given the multifaceted ability of MNS to be applied to varying vascular beds, we describe the successful employment of this modality to facilitate treatment of lower extremity peripheral arterial disease in a symptomatic male with gangrenous toe ulceration. PMID- 19133684 TI - Endovascular balloon occlusion for catheter-induced large artery perforation in the catheterization laboratory. AB - BACKGROUND: Vessel perforation is a complication that cannot be completely avoided in the setting of endovascular procedures. When a large noncompressible artery is disrupted, uncontrolled bleeding may lead to hemodynamic collapse. Endovascular occlusion may provide rapid control of hemorrhage and facilitate definitive therapy; yet, occlusion balloons are not commonly utilized in the cardiac catheterization laboratory. METHODS: We describe our experience with the use of dedicated occlusion balloons for the management of catheter-induced major bleeding events. Endovascular occlusion was utilized to control severe bleeding in five patients with perforation of the aorta, iliac, femoral, or pulmonary artery. RESULTS: Acute control of hemorrhage was achieved in all patients with hemodynamic stabilization in four of five cases. Endovascular balloon occlusion facilitated definitive therapy with surgical repair in two patients, covered stent deployment in one patient and transcatheter vessel occlusion in one patient. Care was withdrawn in one elderly patient with multiple comorbidities. All four patients who survived the procedure were discharged alive from the hospital. CONCLUSIONS: In most cases of severe catheter-induced bleeding, endovascular balloon occlusion provide a safe, rapid, and effective means of temporary bleeding control. PMID- 19133685 TI - Treatment of aortic arch aneurysm with a NuMED-covered stent and restoration of flow to excluded left subclavian artery: perforation and dilation of e-PTFE can be done! AB - We describe a case using a NuMED-covered Cheatham-Platinum (CP) stent (NuMED, Hopkinton, NY) to treat an aneurysm after previous balloon angioplasty and bare stent implantation for coarctation of the aorta (CoA). Exclusion of the left subclavian artery (LSCA) was anticipated. After wire perforation of the covered CP stent, balloon angioplasty was performed through a stent cell to recannulize the LSCA. PMID- 19133686 TI - Collagen embolization for the successful treatment of a distal coronary artery perforation. AB - A 42-year old man suffered a coronary artery perforation in the right coronary artery secondary to a hydrophilic guidewire. We describe for the first time the use of collagen embolization to successfully treat this complication. PMID- 19133687 TI - Implantation of two self-expanding aortic bioprosthetic valves during the same procedure-Insights into valve-in-valve implantation ("Russian doll concept"). AB - Transcatheter aortic valve implantation has gained acceptance as a treatment for elderly patients considered high risk for surgical aortic valve replacement. There are still, however, many unresolved clinical and technical issues. The occurrence of transcatheter aortic valve-in-valve implantation has been reported anecdotally. Aside from a single case report, there is little literature on this topic. This study was conducted to evaluate the procedural, imaging, and clinical outcomes of patients who underwent transcatheter valve-in-valve implantation with two self-expanding aortic valve bioprostheses during the same procedure. We discuss also the potential valve of on-line quantitative angiography for assessing the depth of valve implantation and the need to implant a second valve. PMID- 19133688 TI - Novel technique using the Outback LTD catheter for a common iliac artery occlusion. AB - We present a case of an unusual arterial--arterial anastamosis that resulted from an unsuccessful attempt at crossing a total occlusion of the common iliac artery. Subsequently, we were able to successful recanalize the artery using a modified technique with the Outback reentry catheter. PMID- 19133690 TI - Cortisol levels in Prader-Willi syndrome support changes in routine care. PMID- 19133689 TI - Gene expression patterns in experimental colitis in IL-10-deficient mice. AB - BACKGROUND: While others have described gene expression patterns in humans with inflammatory bowel diseases and animals with chemically induced colitis, a genome wide comparison of gene expression in genetically susceptible animals that develop spontaneous colitis has not been reported. METHODS: We used microarray technology to compare gene expression profiles in cecal specimens from specific pathogen-free IL10-deficient (IL10-/-) mice with colitis and normal wildtype (WT) mice. RNA isolated from ceca of IL10-/- and WT mice was subjected to microarray analysis. The results were confirmed by real-time polymerase chain reaction (PCR) and immunofluorescence microscopy of selected molecules. Expression of the selected genes in dextran sodium sulfate (DSS)-treated mice with colitis and epithelial cell lines activated with pathophysiologic stimuli was measured by real-time PCR. RESULTS: Histological inflammation of the colon and IL-12/23p40 secretion from intestinal explants were greater in IL10-/- and DSS-treated mice versus WT and untreated mice. Microarray analysis demonstrated >10-fold induction of the following molecules in the ceca of IL10-/- mice: mitochondrial ribosomal protein-L33, aquaporin-4, indoleamine-pyrrole-2,3-dioxygenase, and MHC class II with 63, 25, 20, and 12-fold increases, respectively. Cytochrome-P450, pancreatic lipase-related protein-2, and transthyretin were downregulated in IL10-/- mice. MHC II was increased throughout the colon, and aquaporin-4 was increased in the basolateral aspect of cecal epithelial cells. MHC II mRNA was increased in epithelial cells treated with IFN-gamma, but not TNF or Toll-like receptor ligands. CONCLUSIONS: Although most upregulated genes in experimental colitis are immune-related, aquaporin-4 and mitochondrial ribosomal protein-L33, which have not been previously associated with inflammation, were most highly upregulated. PMID- 19133691 TI - Rett syndrome and long-term disorder profile. AB - In a cohort of 103 females clinically diagnosed with Rett syndrome (RTT), 91 had a detectable MECP2 mutation. Emphasis on details of natural history facilitated grouping of females with the same MECP2 mutation and the development of so-called disorder profiles. Some examples of disorder profiles of different recurrent MECP2 mutations are discussed. RTT females with the frequently recurrent R133C and R306C missense mutations and those with intragenic deletions in the C terminus of MECP2 deserve more attention in larger studies as their development is different and milder in the long term. RTT females with the T158M missense mutation are often atypical with mainly behavioral characteristics in infancy and childhood but become classic RTT in adolescence after a slower, protracted course. PMID- 19133692 TI - 15q overgrowth syndrome: a newly recognized phenotype associated with overgrowth, learning difficulties, characteristic facial appearance, renal anomalies and increased dosage of distal chromosome 15q. AB - Trisomy and tetrasomy of distal chromosome 15q have rarely been reported. Although most of the described patients have some learning difficulties and are overgrown, the phenotype associated with distal trisomy/tetrasomy 15q is uncertain due to the small numbers of reported cases and the common co-occurrence of additional chromosome deletions in many patients with trisomy 15q. We present five individuals with overgrowth, learning difficulties and increased dosage of distal 15q. Partial trisomy 15q was identified in four of these cases. Two were generated through recombination of a parental pericentric inversion and two were generated through malsegregation of a maternal balanced 14;15 reciprocal translocation. In all four cases the trisomy can be considered "pure" as the 14p and 15p monosomies will exert no phenotypic effect. Partial tetrasomy 15q, as the result of an analphoid supernumerary chromosome derived from an inverted duplication of distal 15q, was identified in the fifth patient. In addition to the overgrowth and learning difficulties, all five had a characteristic facial appearance and three had renal anomalies. The gestalt consists of a long, thin face with a prominent chin and nose. Renal anomalies included renal agenesis, horseshoe kidney, and hydronephrosis. We provide further support for a distinct "15q overgrowth syndrome" caused by either trisomy or tetrasomy resulting in increased dosage of distal 15q. In addition we propose that renal anomalies and a distinctive facial appearance be considered major features of this condition. PMID- 19133693 TI - Cognitive profile of disorders associated with dysregulation of the RAS/MAPK signaling cascade. AB - Mutations in genes coding for transducers participating in the RAS/MAPK pathway have been identified as the molecular cause underlying a group of clinically related developmental disorders with cognitive deficits of variable severity. To determine the spectrum of cognitive defects associated with dysregulation of this signal cascade, we studied the profile of cognitive abilities in patients with mutations affecting the PTPN11, SOS1, HRAS, KRAS, BRAF, RAF1, and MEK1 genes and phenotype-genotype correlations. Our findings support the observation that heterogeneity in cognitive abilities can be at least partially ascribed to the individual affected genes and type of mutation involved. While mutations affecting transducers upstream of RAS were less frequently associated with mental retardation, mutations in downstream components of the pathway were generally associated with a more severe cognitive impairment. Among patients with a heterozygous PTPN11 mutation, the T468M substitution was associated with a mean IQ significantly higher compared to that of individuals carrying the N308D change. Our study provides insights on the range of cognitive abilities in patients with gene mutations causing dysregulation of RAS signaling suggesting that the presence and severity of cognitive involvement can be predicted in part by the gene involved. PMID- 19133694 TI - Molecular cytogenetic characterization of two small supernumerary marker chromosomes derived from chromosome 19. PMID- 19133695 TI - Somatic mosaicism in a patient with Lynch syndrome. AB - Genetic mosaicism is the presence of genetically different cell populations within an individual and can be associated with a milder disease phenotype. We describe a somatic mosaicism in a Lynch syndrome patient with a MLH1 gene mutation (c.1050delA). Since she was the sister of a heterozygous proposita, the mosaicism appeared to be caused by reversion of an inherited mutation and not a de novo mutation. In order to better understand her cancer risk, we tested different tissues to quantify the amount of mutated allele in several districts. The mosaicism was analyzed using DNA sequencing, primer extension, and dHPLC. The MLH1 mutation was present in somatic cells representative of the three embryonic layers and its percentage was > or =80% in both blood and tissues. Since this patient had a relevant quota of mutated cells, a significantly milder phenotype is not expected. PMID- 19133697 TI - Paradoxical association of extensive nevus flammeus together with unilateral lower limb and breast hypoplasia. PMID- 19133698 TI - Posttraumatic neuroma of the radial nerve treated with an autogenous epineural conduit technique. A case report. AB - We present the outcome of the first clinical application of a new technique using an epineural flap to bridge a short nerve defect. A 28-year-old male had suffered a radial nerve laceration at the lower third of the arm, proximal to the brachioradialis branch, 3 weeks before surgery. During surgery, a neuroma-in continuity was excised preserving the epineural sleeve. Two longitudinal epineural flaps were created, one from the proximal and one from the distal nerve stump and used to bridge a 1-cm-long nerve defect. Each epineurium flap was sutured to the intact epineurium of the other side and additionally to each other. An electromagnetic nerve stimulator was used to enhance the nerve regeneration process. Nerve regeneration was followed up for 17 months with excellent functional results. PMID- 19133700 TI - Anxiety sensitivity as a predictor of the clinical course of panic disorder: a 1 year follow-up study. AB - BACKGROUND: There is evidence that negative affect (NA) and anxiety sensitivity (AS) predict the development of anxiety disorders, particularly panic disorder (PD). The main purpose of this study was to examine whether NA and AS will also predict the clinical course of PD. METHODS: Participants were 136 individuals with a DSM-III-R diagnosis of PD (with or without agoraphobia) enrolled in a naturalistic and longitudinal study of anxiety disorders, the Harvard/Brown Anxiety Research Project (HARP). Participants were administered the Anxiety Sensitivity Index and the Negative Affect Scales of the Positive and Negative Affect Schedule-Expanded Form (PANAS-X-NA) and their percentage of time in PD episode was followed for 1 year after the administration of the measures. RESULTS: Multiple regression analyses indicated that AS, but not NA, was a significant predictor of percentage of time in PD episode after controlling for previous time in PD episodes, comorbid depression, other anxiety disorders, and exposure to psychopharmacological and behavioral treatments. As expected, the Physical Concerns subscale of the Anxiety Sensitivity Index had a significant independent contribution in predicting the course of the disorder. CONCLUSIONS: Overall, these findings suggest that AS, as a unique construct, may be predictive of the amount of time patients are in episode of PD. PMID- 19133699 TI - Glucocorticoid inhibition in the treatment of depression: can we think outside the endocrine hypothalamus? AB - BACKGROUND: Major depressive disorder affects a substantial percentage of the U.S. population, and can be highly debilitating. Selective serotonin reuptake inhibitors are commonly prescribed to treat depression, but may not be as effective for more severe or persistent depression. METHODS: The authors review data concerning the effects of corticosteroid synthesis inhibitors (CSIs) in the management of depressive disorders, present a hypothesis as to their possible mechanisms of action based on recent data suggesting synergistic effects of glucocorticoids on extrahypothalamic corticotropin-releasing hormone (CRH), and consider alternative hypotheses. Published reports evaluating the efficacy of CSIs in treating depression are reviewed and presented in light of recent findings regarding actions of glucocorticoids on the central CRH system. RESULTS: Results from open label and double-blind studies by several groups have indicated that CSIs may be efficacious or of adjunctive value in some patients with depression, including those refractory to other agents; however, there is a need for more controlled studies. Several lines of data suggest that the mechanism of action of these agents may not be solely a function of inhibition of adrenal cortisol production. CONCLUSIONS: The authors propose that CSIs may be efficacious in part by reducing glucocorticoid enhancement of CRH action in neurons of the central nucleus of the amygdala and other structures outside the endocrine hypothalamus. Possible effects of systemically administered CSIs on glucocorticoid receptor regulation, neuroactive steroids, and classical monoamine systems are also discussed. We conclude that available clinical data suggest a potential role for CSIs in the management of depressive disorders, especially major depression with psychotic features. PMID- 19133701 TI - Pain interference impacts response to treatment for anxiety disorders. AB - BACKGROUND: Anxiety disorders and pain are commonly comorbid, though little is known about the effect of pain on the course and treatment of anxiety. METHODS: This is a secondary analysis of a randomized controlled trial for anxiety treatment in primary care. Participants with panic disorder (PD) and/or generalized anxiety disorder (GAD) (N=191; 81% female, mean age 44) were randomized to either their primary-care physician's usual care or a 12-month course of telephone-based collaborative care. Anxiety severity, pain interference, health-related quality of life, health services use, and employment status were assessed at baseline, and at 2-, 4-, 8-, and 12-month follow-up. We defined response to anxiety treatment as a 40% or greater improvement from baseline on anxiety severity scales at 12-month follow-up. RESULTS: The 39% who reported high pain interference at baseline had more severe anxiety (mean SIGH-A score: 21.8 versus 18.0, P<.001), greater limitations in activities of daily living, and more work days missed in the previous month (5.8 versus 4.0 days, P=.01) than those with low pain interference. At 12-month follow-up, high pain interference was associated with a lower likelihood of responding to anxiety treatment (OR=.28; 95% CI=.12-.63) and higher health services use (26.1% with >/=1 hospitalization versus 12.0%, P<.001). CONCLUSIONS: Pain that interferes with daily activities is prevalent among primary care patients with PD/GAD and associated with more severe anxiety, worse daily functioning, higher health services use, and a lower likelihood of responding to treatment for PD/GAD. PMID- 19133702 TI - Added benefits and user satisfaction with a low-dose oral contraceptive containing drospirenone: results of three multicentre trials. AB - The ethinylestradiol 20 microg/drospirenone 3 mg combined oral contraceptive (COC), administered in cycles of 21 days continuous use followed by a 7-day hormone-free interval (21/7) [Yasminelle], has been proven to be an effective and well tolerated contraceptive with an acceptable bleeding pattern and good safety profile. In addition, the ethinylestradiol 20 microg/drospirenone 3 mg 21/7 COC prevents water-retention-related weight gain, and improves physical and emotional well-being, consistent with other drospirenone-containing COCs. In clinical studies, >85% of those who received the ethinylestradiol 20 microg/drospirenone 3 mg 21/7 COC were satisfied or very satisfied with this contraceptive. The positive attributes of the ethinylestradiol 20 microg/drospirenone 3 mg 21/7 COC beyond contraceptive protection may have important implications for improving treatment compliance and reducing treatment discontinuation. Physical well-being stayed the same or improved during use of this COC in 56% and 34% of women, respectively. Emotional well-being stayed the same or improved in 71% and 20% of women, respectively. PMID- 19133703 TI - Safety and tolerability of fluticasone furoate nasal spray once daily in paediatric patients aged 6-11 years with allergic rhinitis: subanalysis of three randomized, double-blind, placebo-controlled, multicentre studies. AB - BACKGROUND: Fluticasone furoate is a novel enhanced-affinity corticosteroid that has demonstrated favourable safety and tolerability in paediatric patients. Three pivotal phase III studies were conducted in children as young as 2 years of age; however, because the European Medicines Agency has recently approved fluticasone furoate for use in children aged>or=6 years, a subanalysis has been conducted to support safety and tolerability in the 6- to 11-year age group. METHODS: All three trials included in the subanalysis were randomized, double-blind, placebo controlled, parallel-group studies: a 2-week US study in patients with seasonal allergic rhinitis (SAR), a 12-week global study in patients with perennial allergic rhinitis (PAR) and a 6-week US hypothalamic-pituitary-adrenal (HPA) axis safety study in patients with PAR. Randomized patients received once-daily treatment with either fluticasone furoate nasal spray (FFNS) 55 microg (n=297; 2 week SAR and 12-week PAR studies only), FFNS 110 microg (n=321) or vehicle placebo nasal spray (n=330). Safety assessments included clinical adverse event (AE) monitoring, clinical laboratory tests, detailed nasal examinations, monitoring of vital signs, and 12-lead ECGs. Ophthalmic examinations (12-week PAR study only) were conducted by an ophthalmologist or optometrist before randomization and at final treatment visit. Twenty-four-hour urinary cortisol (UC) excretions (6- and 12-week PAR studies) and serum cortisol (SC) concentrations (6-week PAR study only) were assessed at specified timepoints over 24 hours as a measure of HPA axis function. In the 6-week HPA axis safety study in patients with PAR, 24-hour UC and SC samples were collected in a domiciled (clinical) setting. RESULTS: No unexpected safety results were observed in patients aged 6-11 years enrolled in the three studies. Across all three trials, AEs considered by the investigator to be drug related were observed in 10%, 7% and 8% of patients in the FFNS 55 microg, FFNS 110 microg and placebo groups, respectively. The most common AEs were headache (8%, 9% and 8% in the FFNS 55 microg, FFNS 110 microg and placebo groups, respectively), nasopharyngitis (5%, 6% and 5%, respectively), pharyngolaryngeal pain (5%, 3% and 4%, respectively), epistaxis (4% in both active treatment groups and 4% in the placebo group) and pyrexia (3% in both active treatment groups and 2% in the placebo group). Findings from nasal examinations were similar across the treatment and placebo groups. Ophthalmic examinations found no differences between the treatment groups for mean change from baseline in intraocular pressure (assessed in the 12-week PAR study only), and no posterior subcapsular cataracts were reported in patients in either FFNS-treatment group. In the 6-week HPA axis study, 24-hour SC geometric mean concentrations were similar for FFNS and placebo groups. The lower limit of two-sided 95% confidence interval (CI) for the treatment ratio was greater than the prespecified noninferiority margin of 0.8 (treatment ratio=0.92, 95% CI 0.80, 1.05). CONCLUSIONS: FFNS has a favourable safety and tolerability profile in patients aged 6-11 years with PAR or SAR. PMID- 19133704 TI - Pharmacokinetic profile of armodafinil in healthy subjects: pooled analysis of data from three randomized studies. AB - BACKGROUND AND OBJECTIVES: Armodafinil (R-modafinil) is the R- and longer-lasting isomer of the racemic compound modafinil, a wakefulness-promoting medication. Armodafinil is eliminated approximately three times more slowly than the S-isomer of racemic modafinil. Published studies have demonstrated the efficacy of armodafinil for treating excessive sleepiness associated with obstructive sleep apnoea, shift work disorder and narcolepsy. The objectives of this study were to describe the pharmacokinetic profile, tolerability and safety of armodafinil in healthy subjects. METHODS: Pooled pharmacokinetic data from three separate randomized studies in 119 healthy subjects who received single or multiple (once daily for up to 14 days) oral doses of armodafinil ranging between 50 and 400 mg were analysed. The impact of food on the single-dose pharmacokinetic profile of armodafinil was also assessed in subjects following an overnight fast and after the consumption of a standard fatty meal. RESULTS: Armodafinil was readily absorbed and exhibited linear pharmacokinetics over the 50-400 mg dose range. Peak plasma concentrations were reached around 2 hours after administration in the fasted state. Food had no effect on the overall bioavailability of armodafinil; however, the peak concentration was delayed by approximately 2-4 hours. In the multiple-dose study, dose proportionality was confirmed by linear regression analyses of the log-transformed area under the plasma concentration versus time curve (AUC) and maximum plasma concentration (Cmax) values as a function of dose. After reaching the peak, plasma concentrations of armodafinil declined in a monophasic manner, with a mean elimination half-life of approximately 15 hours. Steady state appeared to be reached within 7 days. At steady state, the systemic exposure to armodafinil was 1.8 times that observed after single-dose administration. Armodafinil was generally well tolerated, the most frequent adverse events being headache, dizziness and nausea. CONCLUSIONS: In the present analysis, armodafinil exhibited linear pharmacokinetics over the dose range of 50-400 mg. While food affected the rate but not the extent of absorption, peak plasma concentrations were reached in approximately 2 hours when the drug was taken on an empty stomach. With once-daily dosing, steady state appeared to be reached within 7 days. After reaching peak plasma levels, concentrations of armodafinil declined monophasically, with a mean elimination half-life of around 15 hours. Armodafinil was generally well tolerated. PMID- 19133705 TI - Long-term evaluation of glucose homeostasis in a cohort of HAART-treated HIV infected children: a longitudinal, observational cohort study. AB - BACKGROUND AND OBJECTIVES: Few and mainly cross-sectional studies of glucose homeostasis are available in HIV-infected children treated with highly active antiretroviral therapy (HAART). The aim of the present study was to describe a 4 year course of glucose homeostasis in a cohort of HAART-treated children and adolescents, using glucose and insulin levels during an oral glucose tolerance test (OGTT) as outcome measures. In addition, we investigated possible risk factors, both related and unrelated to antiretroviral therapy, associated with insulin resistance. METHODS: We assessed glucose metabolism yearly for 4 consecutive years in 37 HIV-infected children receiving a protease inhibitor (PI) based HAART regimen containing lamivudine/stavudine plus indinavir or ritonavir or nelfinavir or a non-nucleoside reverse transcriptase inhibitor (NNRTI)-based HAART regimen containing lamivudine/tenofovir/efavirenz. Generalized estimating equations were used to evaluate the relationship between the loge-transformed area under the serum concentration-time curve (AUC) of insulin during OGTT and antiretroviral therapy, controlling for time, sex, baseline age, puberty, body mass index and CD4+ T cells percentage. RESULTS: Ritonavir-unboosted PI-based HAART regimens were administered to most children at baseline; however, their use decreased during follow-up in favour of an NNRTI-based regimen. The nelfinavir/lamivudine/stavudine (regression coefficient=-0.69, p<0.05) and efavirenz/lamivudine/tenofovir (regression coefficient=-0.93, p<0.05) regimens, but not the ritonavir/lamivudine/stavudine regimen, were negatively associated with loge-transformed insulin AUC compared with indinavir/lamivudine/stavudine. Puberty was positively associated with loge-transformed insulin AUC. CONCLUSIONS: This 4-year prospective study of HAART-treated HIV-infected children shows that: (i) the nelfinavir/lamivudine/stavudine and the efavirenz/lamivudine/tenofovir regimens but not the ritonavir/lamivudine/stavudine regimen were associated with higher insulin sensivity, i.e. lower insulin AUC, compared with indinavir/lamivudine/stavudine; (ii) the treatment switched substantially in favour of NNRTI from the third year on and this change was associated with an improvement in insulin sensitivity compared with the previous HAART-based regimens; and (iii) puberty is a primary determinant of insulin sensitivity. PMID- 19133707 TI - Addition of dexmedetomidine or lornoxicam to prilocaine in intravenous regional anaesthesia for hand or forearm surgery: a randomized controlled study. AB - BACKGROUND AND OBJECTIVES: Intravenous regional anaesthesia (IVRA) is a simple and cost-effective technique that is ideally suited for surgery involving the distal arm. This study compared the effect of lornoxicam or dexmedetomidine in IVRA with prilocaine in patients who underwent hand or forearm surgery. M ethods: This randomized, double-blind study enrolled 75 patients scheduled for hand or forearm surgery. IVRA was achieved with 2% prilocaine 3 mg/kg in the control group (n=25), 2% prilocaine 3 mg/kg plus dexmedetomidine 0.5 microg/kg in the dexmedetomidine group (n=25), and 2% prilocaine 3 mg/kg plus lornoxicam 8 mg in the lornoxicam group (n=25). In all groups, 0.9% NaCl solution was added to make up a total volume of 40 mL. Sensory and motor block onset and recovery times, haemodynamic variables, visual analogue scale (VAS) pain and sedation scores, duration of analgesia, total analgesic consumption over 24 hours, adverse effects and quality of anaesthesia were recorded. RESULTS: Sensory block onset was shorter and sensory block recovery time longer in the dexmedetomidine group compared with the lornoxicam and control groups (p<0.05). Sensory and motor block recovery times and duration of analgesia for tourniquet were prolonged in the dexmedetomidine and lornoxicam groups compared with the control group (p<0.05). Median VAS scores for tourniquet pain in the dexmedetomidine and lornoxicam groups were lower than that of the control group at 15 and 30 minutes (p<0.05). Postoperatively, the duration of analgesia time was longer and median VAS scores were lower during the first 12 hours in the dexmedetomidine and lornoxicam groups compared with the control group (p<0.05). Total analgesic consumption over 24 hours was lower in the dexmedetomidine and lornoxicam groups compared with the control group (p<0.05). Anaesthesia quality as determined by the anaesthesiologist was better in the dexmedetomidine and lornoxicam group than in the control group (p<0.05). CONCLUSIONS: Addition of dexmedetomidine or lornoxicam to prilocaine in IVRA decreased VAS pain scores, improved anaesthesia quality and decreased analgesic requirement. We suggest that addition of dexmedetomidine or lornoxicam at the doses used in this study to IVRA with prilocaine in this setting can be useful without causing adverse effects. No hypotension, bradycardia or hypoxia requiring treatment was seen in any of the patients. Addition of dexmedetomidine had a more potent effect, shortening sensory block onset time and prolonging sensory block recovery time more than lornoxicam. PMID- 19133706 TI - Cost effectiveness of travoprost versus a fixed combination of latanoprost/timolol in patients with ocular hypertension or glaucoma: analysis based on the UK general practitioner research database. AB - OBJECTIVE: This study aimed to compare the cost effectiveness of travoprost versus a fixed combination of latanoprost/timolol as first-line therapies for ocular hypertension or glaucoma. METHODS: Patient charts were extracted from the UK General Practitioner Research Database. Patients with ocular hypertension or glaucoma who received first-line treatment with either travoprost or latanoprost/timolol and were followed up for >6 months were included. Treatment failure was defined as a treatment change or a glaucoma intervention (laser therapy or surgery). Time to treatment failure was compared using a Cox model and adjusted by the propensity score method. RESULTS: Eligible patients received either travoprost (n=639) or latanoprost/timolol (n=176). Their mean age was 70 years at diagnosis and 48.2% of patients were male. Patient characteristics did not differ significantly between treatment groups. Treatment failure rates at 1 year were 31.3% (travoprost) and 39.4% (latanoprost/timolol) and yielded a hazard ratio for failure in favour of travoprost (0.75; p<0.04) after adjusting for age, sex, co-morbidities and duration of follow-up. Adjusted annual costs of glaucoma management were significantly (p<0.001) less with travoprost (pound215.86) than with latanoprost/timolol (pound327.83). CONCLUSIONS: In everyday practice, travoprost was maintained longer than latanoprost/timolol as first-line therapy for glaucoma. The mean daily costs of travoprost were 50.8% less per patient than those of latanoprost/timolol. Despite adjustments, these results might be confounded, at least partially, by disease severity. PMID- 19133708 TI - Use of N-terminal pro-brain natriuretic peptide to assess left ventricular function after adjuvant doxorubicin therapy in early breast cancer patients: a prospective series. AB - BACKGROUND AND OBJECTIVE: Anthracyclines are well established and highly efficacious antineoplastic agents for various haematopoietic and solid tumours, such as breast cancer. The main adverse effect of anthracycline therapy is cardiotoxicity. The aim of this prospective study was to determine the role of plasma levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) in assessing left ventricular function in early breast cancer patients receiving adjuvant anthracycline treatment. METHODS: Thirty-three newly diagnosed breast cancer patients who received a total doxorubicin dosage of 240 mg/m2 over four treatment cycles as part of adjuvant chemotherapy after curative breast surgery were included in this study. Venous NT-proBNP levels were measured before and at the end of doxorubicin therapy. Left ventricular function was measured by echocardiography conducted 3 weeks after surgery and at the end of doxorubicin therapy. RESULTS: NT-proBNP levels were significantly higher in patients (n=10) with decreased left ventricular ejection fraction (LVEF) [p=0.02]. There was no difference in LVEF (p=0.164) or NT-proBNP levels (p=0.844) between the patients who had high NT-proBNP levels and those who had normal NT-proBNP levels before doxorubicin chemotherapy. None of the factors studied (breast cancer grade, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor 2 status, age) was found to be significantly related to NT proBNP. CONCLUSION: The association between higher NT-proBNP levels and reduced LVEF in asymptomatic breast cancer patients after doxorubicin administration could be an early indication of subclinical acute anthracycline cardiotoxicity. Furthermore, breast cancer patients experiencing a progressive increase in NT proBNP levels might be in a higher risk group for acute anthracycline cardiotoxicity. PMID- 19133710 TI - Body mass index and weight gain as alternative or complementary hypothesis to explain olanzapine concentration-dose ratio accumulation. PMID- 19133709 TI - Topiramate in pregnancy and breastfeeding. AB - Recent information suggests that use of the antiepileptic drug topiramate in pregnancy may raise some concerns, especially if used in polytherapy. Moreover, data on the safety of this antiepileptic drug for the breastfed infant are very limited. However, use of topiramate may be unavoidable in women who wish to become pregnant but who have already experienced severe adverse reactions to antiepileptic medications considered relatively safe in this female condition. Hence, we describe the healthy outcome of a male infant exposed to topiramate through the placenta and maternal milk and who was conceived while the mother was undergoing folic acid supplementation. PMID- 19133712 TI - Critical period of sensitivity for effects of cadmium on frog growth and development. AB - Cadmium is a ubiquitous pollutant in aquatic environments that can alter organismal physiology and ecology. Previous experiments found that ecological Cd exposures increased the growth and development of two North American anurans. However, the generality of these effects among species, the time period over which they occur, and the mechanisms responsible remain conjectural. The goal of the present study was to determine the critical period of sensitivity of Rana pipiens exposed to ecologically relevant levels of Cd. We exposed tadpoles to Cd (0 [control], 1.0, and 10.0 microg/L) from Gosner stage (GS) 25 to metamorphic climax. We assessed effects of Cd on amphibian length, survival, and development during premetamorphosis (GS 25-30) and prometamorphosis (GS 31-42). After 14 d of exposure, we staged tadpoles and recorded snout-vent length. Tadpoles were then pooled according to treatment and stage (GS < or =29 or GS > or =30) and allowed to undergo metamorphic development. Tadpoles exposed to 10 microg/L were significantly larger and more advanced in development by 14 d. Survival to forelimb emergence exceeded 90% in all treatments, and time to metamorphic climax was not different from that in controls. Body burdens of Cd were positively correlated with increasing treatment. Early amphibian development (premetamorphosis) was shown to be the critical period of sensitivity for growth and development. Whereas the freshwater criterion for Cd appears to be protective for survival, a lack of knowledge remains about the sublethal effects of chronic exposures of metal pollutants, especially as they relate to tissue concentrations at various stages of amphibian life history. PMID- 19133718 TI - Effect of poly(N-vinyl-pyrrolidone)-block-poly(D,L-lactide) as coating agent on the opsonization, phagocytosis, and pharmacokinetics of biodegradable nanoparticles. AB - The effect of the coating polymer poly(N-vinyl-pyrrolidone) (PVP) on the protein adsorption, phagocytosis, and pharmacokinetics of poly(D,L-lactide)-based nanoparticles was evaluated in vitro and in vivo. Control poly(ethylene glycol) (PEG)-coated nanoparticles were included for comparison. While no difference between PEG- and PVP-decorated nanoparticles in terms of amount of adsorbed protein was evident upon incubation in single protein solutions (BSA, IgG), incubation in serum revealed a protein adsorption pattern both quantitatively and qualitatively distinct. Larger amounts of complement components and immunoglobulins were found to adhere to PVP-coated particles, whereas PEG particles showed preferential adsorption of apolipoproteins. Furthermore, preopsonization in fresh rather than heat-inactivated serum enhanced uptake of both types of particles by murine RAW 264.7 macrophages. However, when isolated rat Kupffer cells were employed, activation of the complement system significantly enhanced the uptake of PVP-coated nanoparticles compared to PEG particles. Ultimately, PVP-coated nanoparticles exhibited considerably shorter circulation times compared to their PEG counterparts when administered intravenously to rats. PMID- 19133717 TI - HPMA polymer-based site-specific delivery of oligonucleotides to hepatic stellate cells. AB - The objective was to determine whether bioconjugation of type I collagen specific triplex forming oligonucleotide (TFO) to N-(2-hydroxypropyl) methacrylamide (HPMA) containing tetrapeptide Gly-Phe-Leu-Gly (GFLG) and mannose 6-phosphate (M6P) can provide their targeted delivery to hepatic stellate cells (HSCs). Following bioconjugation, M6P-GFLG-HPMA-GFLG-32P-TFO was characterized by PAGE, HPLC, and GPC, and then its biodistribution was determined. TFO was dissociated from the conjugate when incubated with papain and formed triplex with the target DNA duplex. Type 1 collagen gene expression was significantly inhibited when HSC T6 cells were transfected with this conjugate. Following tail vein injection into rats, M6P-GFLG-HPMA-GFLG-(32)P-TFO was rapidly cleared from the circulation and accumulated mainly in the liver. The plasma concentration versus time profile was biphasic, with 12.37 min as t(1/2) of distribution and 2886.48 min as t(1/2) of elimination. A large proportion of the injected M6P-GFLG-HPMA-GFLG-32P-TFO was taken up by the HSCs of both normal and fibrotic rats, which were isolated by liver perfusion at 30 min post-injection. Preinjection of M6P-GFLG-HPMA-GFLG-ONP into fibrotic rats decreased the liver uptake of the conjugates from 60% to 13%, suggesting M6P/TGFII receptor-mediated endocytosis of the conjugates by HSCs. Almost 80% of the total liver uptake in fibrotic rats was contributed by HSCs. In conclusion, conjugation with M6P-HPMA-GFLG significantly increased TFO delivery to the HSCs and could be potentially used for treating liver fibrosis. PMID- 19133719 TI - Influence of the lipidation motif on the partitioning and association of N-Ras in model membrane subdomains. AB - In a combined chemical biological and biophysical approach using time-lapse tapping-mode atomic force microscopy, we studied the partitioning of differently lipidated N-Ras proteins with various membrane-localization motifs into lipid domains of canonical model raft mixtures. The results provide direct evidence that partitioning of N-Ras occurs preferentially into liquid-disordered lipid domains, independent of the lipid anchor system. N-Ras proteins bearing at least one farnesyl group have a comparable membrane partitioning behavior and show diffusion of the protein into the liquid-disordered/liquid-ordered phase boundary region, thus leading to a decrease of the unfavorable line tension between domains. In addition, except for the monofarnesylated N-Ras, strong intermolecular interactions foster self-association and formation of nanoclusters at the domain boundaries and may serve as an important vehicle for association processes and nanoclustering, which has also been observed in in vivo studies. No significant changes of the localization between GDP- and GTP-loaded N-Ras could be detected. Conversely, the nonbiological dual-hexadecylated N-Ras exhibits a time-independent incorporation into the bulk liquid-disordered phase to maintain high conformational entropy of its lipid chains. PMID- 19133721 TI - Fluorescence dynamics of directly meso-meso linked porphyrin rings probed by single molecule spectroscopy. AB - Porphyrin rings CZ4, CZ6, and CZ8 that respectively comprise four, six, and eight porphyrins, immobilized in a thin PMMA film, have been investigated using single molecule fluorescence spectroscopy with a focus on the influences of the overall structural rigidity as the ring size of porphyrin array increases. Neighboring porphyrin moieties were linked directly to enhance through-bond electronic interactions and, as a consequence, efficient excitation energy migration processes like the natural LH2 complex. Unlike the ensemble study, the single molecule study using confocal microscopy could eliminate the averaging effect, and consequently provide detailed information on individual molecular behaviors. Indeed, in solution, as a dihedral angle between neighboring porphyrins decreases in the order of CZ6 > CZ8 > CZ4, red-shifted Q-absorption bands and faster excitation energy hopping rates were observed. However, at the single molecule level, we found that they show longer survival times, less frequent on-off behaviors, narrower fluorescence lifetime distributions, and high relative single molecular brightness in the order of CZ8 > CZ6 > CZ4, by recording fluorescence intensity trajectories. Especially, CZ4 reveals high photostability with its rigid structure, and about 3 porphyrin units among the 4 chromophores-constituted molecule behave as a collective coherent domain. Thus, our results single out CZ4 as a potential and promising candidate for application in artificial light harvesting solid-state devices. PMID- 19133720 TI - Entrapment of hydrophobic drugs in nanoparticle monolayers with efficient release into cancer cells. AB - Gold nanoparticles functionalized with water-soluble zwitterionic ligands form kinetically stable complexes with hydrophobic drugs and dyes. These drugs and dyes are efficiently released into cells, as demonstrated through fluorescence microscopy and cytotoxicity assays. Significantly, there is little or no cellular uptake of particle, making these low toxicity particles promising for delivery applications. PMID- 19133722 TI - Comparison of three commercially available DIGE analysis software packages: minimal user intervention in gel-based proteomics. AB - The success of high-performance differential gel electrophoresis using fluorescent dyes (DIGE) depends on the quality of the digital image captured after electrophoresis, the DIGE enabled image analysis software tool chosen for highlighting the differences, and the statistical analysis. This study compares three commonly available DIGE enabled software packages for the first time: DeCyder V6.5 (GE-Healthcare), Progenesis SameSpots V3.0 (Nonlinear Dynamics), and Dymension 3 (Syngene). DIGE gel images of cell culture media samples conditioned by HepG2 and END2 cell lines were used to evaluate the software packages both quantitatively and subjectively considering ease of use with minimal user intervention. Consistency of spot matching across the three software packages was compared, focusing on the top fifty spots ranked statistically by each package. In summary, Progenesis SameSpots outperformed the other two software packages in matching accuracy, possibly being benefited by its new approach: that is, identical spot outline across all the gels. Interestingly, the statistical analysis of the software packages was not consistent on account of differences in workflow, algorithms, and default settings. Results obtained for protein fold changes were substantially different in each package, which indicates that in spite of using internal standards, quantification is software dependent. A future research goal must be to reduce or eliminate user controlled settings, either by automatic sample-to-sample optimization by intelligent software, or by alternative parameter-free segmentation methods. PMID- 19133724 TI - The second golden age of glycomics: from functional glycomics to clinical applications. PMID- 19133723 TI - Templated spherical high density lipoprotein nanoparticles. AB - We report the synthesis of high density lipoprotein (HDL) biomimetic nanoparticles capable of binding cholesterol. These structures use a gold nanoparticle core to template the assembly of a mixed phospholipid layer and the adsorption of apolipoprotein A-I. These synthesized structures have the general size and surface composition of natural HDL and, importantly, bind free cholesterol (K(d) = 4 nM). The determination of the K(d) for these particles, with respect to cholesterol complexation, provides a key starting and comparison point for measuring and evaluating the properties of subsequently developed synthetic versions of HDL. PMID- 19133725 TI - PNIPAM gel-coated gold nanorods for targeted delivery responding to a near infrared laser. AB - Gold nanorods can be used as photothermal converters, permitting near-infrared (NIR) light to be transmitted deep into tissues without causing damage. We prepared hybrid nanorods with a core-shell structure, i.e., a single gold nanorod encapsulated in a poly (N-isopropylacrylamide) nanogel. Hybrid nanorods demonstrated remote, reversible, pulsatile phase transition and in vivo action after irradiation using a NIR laser. PMID- 19133726 TI - Subsurface oxidation for micropatterning silicon (SOMS). AB - Here we present a straightforward patterning technique for silicon: subsurface oxidation for micropatterning silicon (SOMS). In this method, a stencil mask is placed above a silicon surface. Radio-frequency plasma oxidation of the substrate creates a pattern of thicker oxide in the exposed regions. Etching with HF or KOH produces very shallow or much higher aspect ratio features on silicon, respectively, where patterning is confirmed by atomic force microscopy, scanning electron microscopy, and optical microscopy. The oxidation process itself is studied under a variety of reaction conditions, including higher and lower oxygen pressures (2 and 0.5 Torr), a variety of powers (50-400 W), different times and as a function of reagent purity (99.5 or 99.994% oxygen). SOMS can be easily executed in any normal chemistry laboratory with a plasma generator. Because of its simplicity, it may have industrial viability. PMID- 19133728 TI - Confined self-assembly of toric focal conic domains (the effects of confined geometry on the feature size of toric focal conic domains). AB - A smectic liquid crystal (LC) containing a rigid biphenyl group and semifluorinated chains exhibits a high density of toric focal conic domains (TFCDs) arranged in an ordered array when confined within a microchannel. The formation of the TFCDs is strongly influenced by the width (W) and depth (h) of the confined microchannels, most importantly, by the channel depth. We studied a broad variety of microchannels, with varying width in the range of 3-200 mum and depth in the range of 2-10 mum. The radius of the TFCDs increases with increases in the width until the saturated radius is achieved, which is determined by the depth of the channel. We used the elastic-anchoring model of TFCD formation to explain the experimental observations. The model allows one to trace the dependence of the TFCD radius on the channel depth h, to explain why the TFCDs do not form in channels that are too shallow or too narrow. PMID- 19133729 TI - A highly regioselective ring-opening metathesis-cross metathesis process modulated by the electronic effects of the cross metathesis partner: an entry to quaternary prolines. AB - A modulation of the regioselectivity in the ring-opening metathesis-cross metathesis (ROCM) process, originated by the electronic features of the acyclic olefin, is described. Electron-poor and electron-rich olefins showed opposing behavior giving different regioisomers. The reaction opens the way to the synthesis of interesting proline analogues incorporating a quaternary stereocenter. PMID- 19133731 TI - Molecular dynamical approach to the conformational transition in peptide nanorings and nanotubes. AB - We study the conformational transition in d,l-peptide nanorings (PNRs) and nanotubes (PNTs) computationally based on the total energy calculation. Ab initio energy calculation has been carried out to investigate the static states of PNRs, whereas the molecular dynamics (MD) calculation has been employed to examine PNRs' dynamical states. We, then, discuss the time-dependent (TD) feature via the transition process from E-type to B-type and vice versa. The conformational transition occurs easily from E-type equatorial (Eeq) to B-type axial (Bax) but is unreversible for the opposite direction because of a larger activation energy. The TD tracing of the two dihedral angles in the individual amino acid residues reveals that the conformational change propagates along the peptide skeleton ring nearly at the sound velocity. We further expand our study to the tubular forms and reveal that the PNT has an ability to produce the two kinds of homogeneous tubes, being composed of E rings (E-tube) and of B rings (B-tube), and also that these two PNRs should be mixed to produce a binary alloyed PNT. PMID- 19133732 TI - Application of a rhodium-catalyzed addition/cyclization sequence toward the synthesis of polycyclic heteroaromatics. AB - A cascade rhodium-catalyzed addition/cyclization reaction of bifunctional heteroromatic boronate esters to strained bicyclic alkenes has been developed. This method provides an efficient route to generate a variety of polycyclic heteroaromatic molecules containing benzothiophene, benzofuran, and indole moieties. PMID- 19133733 TI - Numerical investigation of photochemical behavior in bromate-1,4-cyclohexanedione reactions. AB - In this study, photocontrolled reaction behaviors in the uncatalyzed and catalyzed bromate-1,4-cyclohexanedione (bromate-CHD) oscillators were investigated numerically. The simulation was performed with two existing models modified to include a photoreaction step denoting the photoreduction of 1,4 benzoquinone in aqueous solution. The reproduced phenomena include photoinduced and photoquenched chemical oscillations. The scene that light with a moderate intensity favors while strong illumination quenches these spontaneous oscillations is also achieved in the simulation. The modified model also illustrates that, as the reaction evolves in time, the influence of light transforms from inhibitory to constructive. The remarkable agreement between simulation and experiment strongly suggests that 1,4-benzoquinone plays a key role in the photoreaction dynamics of the studied systems. PMID- 19133734 TI - First principles study on the diffusion of alkali-metal ions on the armchair single-wall nanotubes. AB - In this paper we have performed density-functional study on the adsorption and diffusion of various alkali-metal ions on the surface of pristine and defective armchair single-wall carbon nanotubes. In the pristine SWNT system, the position above the hexagon is believed to be the most stable site for adsorption, while the adsorption is enhanced in the defective SWNT. In pristine SWNT all the ions prefer to diffuse along the axial direction, with low barriers less than 0.25 eV. In defective SWNT, the axial diffusion is also energetically most preferable, and the barriers increase only slightly and have little influence on the diffusion as compared to pristine SWNT. PMID- 19133735 TI - Ranking the lacunary (Bu4N)9[H[alpha2-P2W17O61]] polyoxometalate's stabilizing ability for Ir(0)(n) nanocluster formation and stabilization using the five criteria method plus necessary control experiments. AB - The primary goal of the present studies is to rank the monoprotonated, lacunary Wells-Dawson-type polyoxometalate [H[alpha2-P2W17O61]](9-) as a stabilizing anion for the formation and subsequent stabilization of Ir(0)(n) nanoclusters in acetone using the five-criteria method developed previously ( Ozkar , S. ; Finke , R. G. J. Am. Chem. Soc. 2002 , 124, 5796 ). A related goal is to compare this potentially tetradentate, three W-O(-) plus one W-OH ligand [H[alpha2 P2W17O61]](9-) system to the nanocluster formation and stabilization abilities of the present "gold standard" polyoxometalate, [P2W15Nb3O62]9-, with its established tridentate, three Nb-O-Nb ligating system. A comprehensive table of 54 references at present examining polyoxometalates (POMs) as additives/stabilizers of nanoclusters is also provided as Table S1 of the Supporting Information . To accomplish the above-noted two main goals, the organic-solvent-soluble tetrabutylammonium salts, (Bu4N)8.4[H1.6[alpha2 P2W17O61]] x 1.4H2O, (Bu4N)9[H[alpha2-P2W17O61]], and (Bu4N)9[P2W15Nb3O62] were prepared and their basic structures (by IR) and purity (by 31P NMR, plus elemental analysis where appropriate) were determined. The parent Wells-Dawson POM (Bu4N)6[alpha-P2W18O62] was also prepared, characterized, and then used as a control of a polyoxometalate with little surface anionic charge density, one therefore expected to be a poor stabilizer for at least metal(0)(n), overall neutral core, nanoclusters. Also prepared and characterized was (Bu4N)4[H3[PW11O39]], but its attempted deprotonation by (Bu4N)OH with direct 31P NMR monitoring did not yield a clean product by 31P NMR, so studies with this second lacunary POM were deemphasized. The resultant POMs of known composition, protonation state, and thus overall charge were then evaluated by the five criteriathe one presently available methodfor their ability to promote the kinetically controlled formation, stabilization, and subsequent catalytic activity of prototype Ir(0)(n) nanoclusters. A number of additional control experiments necessary to provide confidence in the results are also reported. One main finding is that the efficacy of the POMs studied herein, as stabilizers for Ir(0)(n) nanoclusters in acetone solvent, is [P2W15Nb3O62]9- > [H[alpha2 P2W17O61]]9- >> [alpha-P2W18O62]6- approximately [H3[PW11O39]]4-, the potentially tetradentate, lacunary [H[alpha2-P2W17O61]]9-proving somewhat less efficacious as a stabilizer than the tridentate Nb-O-Nb containing [P2W15Nb3O62]9-POM. Another finding is that the degree of protonation and the overall charge of the POM matter, the more highly charged [H[alpha2-P2W17O61]]9- POM being a better stabilizer then the more protonated, less charged [H2[alpha2-P2W17O61]]8-. Two additional important findings are better insights into the inherent errors underlying three of the five criteria, and thus the use of the five-criteria method itself, and further evidence supporting the hypothesis that future studies of direct measurements of nanocluster agglomeration rate constants k3 and k4 ( Ott , L. S. ; Finke , R. G. Chem. Mater. 2008 , 20, 2592 - 2601 ) should prove valuable. PMID- 19133736 TI - Self-assembly of nanoparticle arrays on semiconductor substrate for charge transfer cascade. AB - The current study focuses on the interaction between hierarchical structures of nanoparticles (NPs) and a semiconductor substrate on which they are assembled. Monolayer and bilayer assemblies of two different NPs were prepared on the surface of a GaAs substrate. The photoluminescence response of the bilayer assemblies depends on their hierarchy, namely on the ordering of differently sized nanoparticles with respect to the surface; however, the surface photovoltage does not. Based on these studies, it is possible to determine the importance of each of the possible quenching mechanisms for electron-hole pair excitation. PMID- 19133737 TI - Toward understanding the conformational dynamics of RNA ligation. AB - Members of the genus Trypanosoma, which include the pathogenic species Trypanosoma brucei and Trypanosoma cruzi, edit their post-transcriptional mitochondrial RNA via a multiprotein complex called the editosome. In T. brucei, the RNA is nicked prior to uridylate insertion and deletion. Following editing, nicked RNA is religated by one of two RNA-editing ligases (TbREL). This study describes a recent 70 ns molecular dynamics simulation of TbREL1, an ATP dependent RNA-editing ligase of the nucleotidyltransferase superfamily that is required for the survival of T. brucei insect and bloodstream forms. In this work, a model of TbREL1 in complex with its full double-stranded RNA (dsRNA) substrate is created on the basis of the homologous relation between TbREL1 and T4 Rnl2. The simulation captures TbREL1 dynamics in the state immediately preceding RNA ligation, providing insights into the functional dynamics and catalytic mechanism of the kinetoplastid ligation reaction. Important features of RNA binding and specificity are revealed for kinetoplastid ligases and the broader nucleotidyltransferase superfamily. PMID- 19133738 TI - Synthesis of substituted 5-(1,2,4-oxadiazol-5-yl)-3,4-dihydropyrimidine-2(1H) thiones. AB - We have developed a liquid-phase route for combinatorial synthesis of novel substituted 5-(1,2,4-oxadiazol-5-yl)-3,4-dihydropyrimidine-2(1H)-thiones. Biginelli-type three-component condensation of 1-(3-aryl-1,2,4-oxadiazol-5 yl)acetones, thiourea, and benzaldehydes is shown to result in new 5-(1,2,4 oxadiazol-5-yl)-3,4-dihydropyrimidine-2(1H)-thione heterocyclic system. If salicylaldehydes are used in this reaction, a mixture of 5-(1,2,4-oxadiazol-5-yl) 3,4-dihydropyrimidine-2(1H)-thiones and 11-(1,2,4-oxadiazol-5-yl)-2,3,5,6 tetrahydro-4H-2,6-methano-1,3,5-benzoxadiazocine-4-thiones is formed. PMID- 19133739 TI - Temperature-driven mixing-demixing behavior of binary mixtures of the ionic liquid choline bis(trifluoromethylsulfonyl)imide and water. AB - The ionic liquid (2-hydroxyethylammonium)trimethylammonium) bis(trifluoromethylsulfonyl)imide (choline bistriflimide) was obtained as a supercooled liquid at room temperature (melting point=30 degrees C). Crystals of choline bistriflimide suitable for structure determination were grown from the melt in situ on the X-ray diffractometer. The choline cation adopts a folded conformation, whereas the bistriflimide anion exhibits a transoid conformation. The choline cation and the bistriflimide anion are held together by hydrogen bonds between the hydroxyl proton and a sulfonyl oxygen atom. This hydrogen bonding is of importance for the temperature-dependent solubility properties of the ionic liquid. Choline bistriflimide is not miscible with water at room temperature, but forms one phase with water at temperatures above 72 degrees C (equals upper critical solution temperature). 1H NMR studies show that the hydrogen bonds between the choline cation and the bistriflimide anion are substantially weakened above this temperature. The thermophysical properties of water-choline bistriflimide binary mixtures were furthermore studied by a photopyroelectric technique and by adiabatic scanning calorimetry (ASC). By photothermal analysis, besides highly accurate values for the thermal conductivity and effusivity of choline bistriflimide at 30 degrees C, the detailed temperature dependence of both the thermal conductivity and effusivity of the upper and lower part of a critical water-choline bistriflimide mixture in the neighborhood of the mixing-demixing phase transition could be determined with high resolution and accuracy. Together with high resolution ASC data for the heat capacity, experimental values were obtained for the critical exponents alpha and beta, and for the critical amplitude ratio G+/G-. These three values were found to be consistent with theoretical expectations for a three dimensional Ising-type of critical behavior of binary liquid mixtures. PMID- 19133740 TI - Complexation of trivalent lanthanides with planar tridentate aromatic ligands tuned by counteranions and steric constraints. AB - Among the plethora of parameters controlling the stability and structures of lanthanide coordination complexes, it is often difficult to decipher their relative importance in the global complexation processes. The combination of the bond valence method (for analyzing solid state structures) with the thermodynamic site binding model (for unravelling complexation reactions occurring in solution) appears to be an efficient tool for specifically addressing interligand effects, which affect the output of the coordination process. When applied to the reaction of the tridentate aromatic scaffolds 2,2':6',2''-terpyridine (L1) and 2,6 bis(benzimidazol-2-yl)pyridine (L2) with trivalent lanthanides, Ln(III), we demonstrate that the successive fixation of ligands, eventually leading to the triple-helical complexes [Ln(Lk)3]3+, is anticooperative both in the solid state and in solution, with a special sensitivity to the nature of the counteranion and to the peripheral substitution for L2. Consequently, in addition to the classical entropic driving forces resulting from the use of specific metal/ligand ratio, the stoichiometry of the final complex can be tuned by a judicious choice of interligand interactions, as exemplified by the unusual isolation of stable complexes with Ln/L = 2:3 ratios. PMID- 19133741 TI - Modulation in the solute location in block copolymer-surfactant supramolecular assembly: a time-resolved fluorescence study. AB - Effect of cosurfactant concentration on the location of a dissolved solute in a block copolymer-surfactant supramolecular system has been investigated using time resolved fluorescence anisotropy and dynamic Stokes' shift measurements. Pluronic F88 and cosurfactant CTAC have been used to form a supramolecular assembly. The anion of coumarin 343 dye has been used as the solute/probe. It is seen that as the CTAC concentration is increased in the F88-CTAC supramolecular assembly, the microviscosity around the probe gradually increases. The result suggests that the probe undergoes a gradual migration from micellar surface to the interior of the micelle as the concentration of the CTAC is increased. This is also supported by the dynamic Stokes' shift results. It is seen that as the CTAC concentration is increased in the system, the observed Stokes' shift gradually increases due to the movement of the probe away from the bulk water. By comparing the present results with those reported in another pluronic-surfactant system, namely, P123 CTAC, it is indicated that the extent of modulation in the position of the probe in such supramolecular systems is largely determined by the composition of the pluronic, especially on the length of its hydrophilic ethyleneoxide block. PMID- 19133742 TI - Molecular recognition of redox-switchable bis-crown moieties assembled on a dicobalt complex. AB - A new dinuclear complex of cobalt(III) having double bis-crown moieties (1) was synthesized in a one-pot multistep reaction between the dinucleating ligand (H(2)L(1)), Co(OAc)(2) x 4 H(2)O, and 4'-aminomethylbenzo-15-crown 5-ether under aerobic conditions. The complex was characterized by UV-vis, IR, NMR, and CSI mass spectroscopies, and the axial coordination of the 4'-aminomethylbenzo-15 crown 5-ether on both cobalt centers was confirmed. The bis-crown moieties of 1 acted as a potassium ion binding site with a large association constant (K) of 1.97 x 10(5) M(-1) (1:1) caused by the preorganized bis-crown structure. Solvent extraction of K(+) (potassium picrate) with 1 was investigated by UV-vis and NMR, and the extracted K(+) was quantified by an atomic absorption/flame emission spectrophotometer. The cyclic voltammogram of 1 showed irreversible redox waves at -0.57 V and +0.16 V versus Ag/AgCl in DMF, which are ascribed to the Co(III) and Co(II) redox couple. The large peak separation (Delta0.73 V) of this redox couple indicates the association and dissociation of the axial amine on the cobalt centers. The irreversible reduction peak of 1 at -0.57 V was shifted in the positive direction by 80 mV in the presence of the potassium ion, showing the complexation of 1 with the potassium ion. The reversible formation and decomposition of the double bis-crown moieties in 1 caused by the electrical stimulus were also investigated by thin-layer UV-vis spectroelectrochemistry. The switching of the double bis-crown structure of 1 was established over four trials with response to electrical energy. PMID- 19133743 TI - Ligand-free palladium-catalyzed direct arylation of thiazoles at low catalyst loadings. AB - Ligand-free Pd(OAc)2 was found to catalyze very efficiently the direct arylation of thiazole derivatives under very low catalyst concentration. By using activated aryl bromides, the reaction can be performed employing as little as 0.1-0.001 mol % catalyst. With such substrates, this procedure is economically and environmentally attractive. On the other hand, in the presence of more challenging substrates, such as some strongly deactivated or highly congested aryl bromides, in some cases, disappointing results were obtained. PMID- 19133744 TI - Characterizing slight structural disorder in solids by combined solid-state NMR and first principles calculations. AB - A general approach for structural interpretation of local disorder in partially ordered solids is proposed, combining high-resolution two-dimensional (2D) nuclear magnetic resonance (NMR) and first principles calculations. We show that small chemical shift variations of the order of a ppm can be interpreted in detailed structural terms with advanced density functional theory methods. Focusing on a model system of bisphosphinoamine, we demonstrate that the existence and the spatial range of small amplitude disorder can be probed using quantitative statistical analyses of 2D NMR line shapes obtained from through space correlation experiments collected using variable mixing times. We show how low-energy vibration modes calculated from first principles can be conveniently used not as a cause of disorder but, instead, to generate a basis set of physically plausible local distortions to describe candidate static distributions of local geometries. Calculations of (31)P NMR isotropic chemical shifts are then used for the first time to simulate 2D correlation lineshapes associated with these distortions, which permit their evaluation as a potential source of disorder by comparison to experimental 2D cross-peaks between phosphorus sites. This new type of structural constraints allows the identification of changes in the bonding geometry that most likely contribute to the local structural disorder. We thus identify at least one type of structural deformation that is compatible with the experimental 2D NMR data and is also within the order of magnitude of the "thermal ellipsoids" associated with the uncertainties on the atomic positions of the X-ray diffraction structure. PMID- 19133745 TI - Temperature-dependent transitions between normal and inverse isotope effects pertaining to the interaction of H-H and C-H bonds with transition metal centers. AB - Deuterium kinetic isotope effects (KIEs) serve as versatile tools to infer details about reaction mechanisms and the nature of transition states, while equilibrium isotope effects (EIEs) associated with the site preferences of hydrogen and deuterium enable researchers to study aspects of molecular structure. Researchers typically interpret primary deuterium isotope effects based on two simple guidelines: (i) the KIE for an elementary reaction is normal (k(H)/k(D) > 1) and (ii) the EIE is dictated by deuterium preferring to be located in the site corresponding to the highest frequency oscillator. In this Account, we evaluate the applicability of these rules to the interactions of H-H and C-H bonds with a transition metal center. Significantly, experimental and computational studies question the predictability of primary EIEs in these systems based on the notion that deuterium prefers to occupy the highest frequency oscillator. In particular, the EIEs for (i) formation of sigma complexes by coordination of H-H and C-H bonds and (ii) oxidative addition of dihydrogen exhibit unusual temperature dependencies, such that the same system may demonstrate both normal (i.e., K(H)/K(D) > 1) and inverse (i.e., K(H)/K(D) < 1) values. The transition between a normal and inverse EIE indicates that these systems do not demonstrate the typical monotonic variation predicted by the van't Hoff relationship. Instead, the calculated EIEs in these systems are 0 at 0 K, increase to a value greater than 1, and then decrease to unity at infinite temperature. This unusual behavior may be rationalized by considering the individual factors that contribute to the EIE. Specifically, the EIE may be expressed in the form EIE = SYM x MMI x EXC x ZPE (where SYM is the symmetry factor, MMI is the mass-moment of inertia term, EXC is the excitation term, and ZPE is the zero-point energy term), and the distinctive temperature profile results from the inverse ZPE (enthalpy) and normal [SYM x MMI x EXC] (entropy) components opposing each other and having different temperature dependencies. At low temperatures, the ZPE component dominates and the EIE is inverse, while at high temperatures, the [SYM x MMI x EXC] component dominates and the EIE is normal. The inverse nature of the ZPE term is a consequence of the rotational and translational degrees of freedom of RH (R = H, CH(3)) becoming low-energy isotopically sensitive vibrations in the product, while the normal nature of the [SYM x MMI x EXC] component results from deuterium substitution having a larger impact on the moment of inertia of the smaller molecule. PMID- 19133746 TI - Polymer nanoparticles: shape-directed monomer-to-particle synthesis. AB - Well-defined dumbbell and tripartite organic nanoparticles (30-60 nm) were produced via a one-pot direct synthesis of branched amphiphilic block copolymers, avoiding the need for postsynthesis self-assembly steps. We show the mechanism of dumbbell formation is largely a concerted process of particle growth during polymerization, although data suggest that particle-particle linking also occurs, particularly at higher monomer conversions. Dumbbell particles formed using a disulfide bifunctional initiator lead to cleavable structures, underlining the role of initiator functionality in shape control and the potential for functionality placement. Trifunctional initiators allow the direct one-pot synthesis of "tripartite" clover-leaf shaped nanoparticles which would be difficult to achieve through conventional synthesis/self-assembly/cross-linking strategies. PMID- 19133747 TI - The monocarboxylate transporter 8 and L-type amino acid transporters 1 and 2 are expressed in mouse skeletons and in osteoblastic MC3T3-E1 cells. AB - BACKGROUND: Several plasma membrane transporters have been shown to mediate the cellular influx and/or efflux of iodothyronines, including the sodium-independent organic anion co-transporting polypeptide 1 (OATP1), the sodium taurocholate co transporting polypeptide (NTCP), the L-type amino acid transporter 1 (LAT1) and 2 (LAT2), and the monocarboxylate transporter 8 (MCT8). The aim of this study was to investigate if the mRNAs of these transporters were expressed and regulated by thyroid hormone (TH) in mouse calvaria-derived osteoblastic MC3T3-E1 cells and in the fetal and postnatal bones of mice. METHODS: The mRNA expression of the iodothyronine transporters was investigated with real-time polymerase chain reaction analysis in euthyroid and hypothyroid fetuses and litters of mice and in MC3T3-E1 cells treated with increasing doses of triiodothyronine (T(3); 10(-10) to 10(-6) M) or with 10(-8) M T(3) for 1-9 days. RESULTS: MCT8, LAT1, and LAT2 mRNAs were detected in fetal and postnatal femurs and in MC3T3-E1 cells, while OATP1 and NTCP mRNAs were not. LAT1 and LAT2 mRNAs were not affected by TH status in vivo or in vitro or by the stage of bone development or osteoblast maturation (analyzed by the expression of osteocalcin and alkaline phosphatase, which are key markers of osteoblastic differentiation). In contrast, the femoral mRNA expression of MCT8 decreased significantly during post-natal development, whereas MCT8 mRNA expression increased as MC3T3-E1 cells differentiated. We also showed that MCT8 mRNA was up-regulated in the femur of hypothyroid animals, and that it was down-regulated by treatment with T(3) in MC3T3-E1 cells. CONCLUSIONS: This is the first study to demonstrate the mRNA expression of LAT1, LAT2, and MCT8 in the bone tissue of mice and in osteoblast-like cells. In addition, the pattern of MCT8 expression observed in vivo and in vitro suggests that MCT8 may be important to modulate TH effects on osteoblast differentiation and on bone development and metabolism. PMID- 19133748 TI - Time to testing and accessing care among a population of newly diagnosed patients with HIV with a high proportion of Canadian Aboriginals, 1998-2003. AB - Early HIV diagnosis and treatment are important for decreasing HIV transmission and morbidity. By using initial CD4 counts and time to first viral load test, we examined the stage of disease at the time of diagnosis and the time to accessing medical care after diagnosis, respectively. Initial CD4 count, first HIV viral load test, demographics and exposure risks were obtained for all newly diagnosed HIV cases in Northern Alberta from 1998-2003. Time to accessing care was determined as the time between diagnosis and the first viral load test. Correlates were determined using simple descriptive statistics and survival analysis methods. Of 526 HIV cases, median age was 36 years (interquartile range [IQR]: 31-43), 69% were males and 41% were Aboriginal. At diagnosis, 28% of the population had CD4 counts less than 200 cells=mm3. After diagnosis, 92.2% accessed care and median time to care for the entire population was 29 days. In multivariate analysis, age at diagnosis less than 45 years was independently associated with longer median time to care (versus age 45 years or more; adjusted hazard ratio [AHR]: 0.69; 95% confidence interval [CI] 0.55-0.88), while Aboriginal ethnicity (versus Caucasian; AHR: 0.82; 95% CI 0.68-1.01), and nonmetropolitan residence (versus metropolitan; AHR: 0.81; 95% CI 0.65-1.00) were marginally significant correlates for longer times to care. Although more than one quarter of cases were diagnosed at relatively advanced stages of infection, the majority of new HIV cases in Northern Alberta accessed care within 2 months of diagnosis. We need to explore new strategies to facilitate and promote earlier access to testing among individuals at risk. PMID- 19133749 TI - Factors associated with psychotherapy longevity among HIV-positive patients. AB - Mental health concerns are salient to many HIV-positive individuals, yet, there are also numerous challenges with regard to access and retention in care. The present study sought to investigate factors that are associated with longevity in outpatient psychological care for HIV-positive patients. We aimed to better understand what factors (e.g., race, gender, age, history of mental health service utilization) are associated with the length of participation in psychotherapy in our clinic. This is an archival study consisting of chart review of our clinical documentation related to the psychological services provided in our clinic from January 2005 through December 2007. Eligible charts included all HIV-positive individuals seeking psychotherapy in the Department of Psychiatry and Health Behavior at the Medical College of Georgia (n = 87 records; 33.3% female; 52.9% ethnic minority, 57% sexual minority, middle-aged (42.0 years +/- 9.6 years). Results indicated that the presence of cohabitating social support distinguished between those who continued or terminated after the intake session, with social support being associated with continuation after intake. Furthermore, shorter distance from clinic, history of previous psychotherapy, concurrent pharmacotherapy, comorbid personality disorder diagnosis, and having an ethnic minority provider were associated with greater longevity in treatment. As a result of modest sample size relative to the number of predictor variables, analyses utilized multiple bivariate analyses and thus, results must be considered preliminary and should be replicated in a larger investigation. However, treatment providers may utilize these initial findings to improve retention in care and to improve the quality of mental health care provided to HIV-positive outpatients. PMID- 19133750 TI - Evaluating health information technology: provider satisfaction with an HIV specific, electronic clinical management and reporting system. AB - Health information technology (HIT) offers the potential to improve care for persons living with HIV. Provider satisfaction with HIT is essential to realize benefits, yet its evaluation presents challenges. An HIV-specific, electronic clinical management and reporting system was implemented in Louisiana's eight HIV clinics, serving over 7500. A serial cross-sectional survey was administered at three points between April 2002 and July 2005; qualitative methods were used to augment quantitative. Multivariable methods were used to characterize provider satisfaction. The majority of the sample (n = 196; T1 = 105; T2 = 46; T3 = 45) was female (80.0%), between ages of 25 and 50 years (68.3%), frequent providers at that clinic (53.7% more than 4 days per week), and had been at the same clinic for a year or more (85.0%). Improvements in satisfaction were observed in patient tracking ( p < 0.05), distribution of educational materials ( p < 0.04), and belief that electronic systems improve care ( p < 0.05). Provider self-reports of time to complete critical functions decreased for all tasks, two significantly so. Time (in minutes) to find current CD4 count decreased at each time point (mean 3.9 [standard deviation {SD} 5.8], 2.9 [2.3], 2.1 [2.6], p>0.05), current viral load decreased at each time point (mean 4.0 [SD 5.6], 2.9 [2.5], 1.8 [2.6], p = 0.08], current antiretroviral status decreased at each time point (mean 3.9 [SD 4.7], 2.9 [3.7], 1.5 [1.1], p < 0.04), history of antiretroviral use decreased at each time point (mean 15.1 [SD 21.9], 6.0 [5.4], 5.4 [7.2], p < 0.04]. Time savings were realized, averaging 16.1 minutes per visit ( p < 0.04). Providers were satisfied with HIT in multiple domains, and significant time savings were realized. PMID- 19133751 TI - Spiritual and mind-body beliefs as barriers and motivators to HIV-treatment decision-making and medication adherence? A qualitative study. AB - We examined spiritual/mind-body beliefs related to treatment decision-making and adherence in 79 HIV-positive people (35% female, 41% African American, 22% Latino, 24% White) who had been offered antiretroviral treatment by their physicians. Interviews (performed in 2003) identified spiritual/mind-body beliefs; the Adult AIDS Clinical Trials Group (ACTG) questionnaire assessed adherence and symptoms/side effects. Decision-making was influenced by health related spiritual beliefs (e.g., calling on God/Higher Power for help/protection, God/Higher Power controls health) and mind-body beliefs (e.g., mind controls body, body tells when medication is needed). Participants believing God/Higher Power controls health were 4.75 times more likely to refuse, and participants with mind-body beliefs related to decision-making were 5.31 times more likely to defer antiretrovirals than those without those beliefs. Participants believing spirituality helps coping with side effects reported significantly better adherence and fewer symptoms/side effects. Fewer symptoms/side effects were significantly associated with the beliefs mind controls body, calling on God/Higher Power for help/protection, and spirituality helps adherence. Spiritual/mind-body beliefs as barriers or motivators to taking or adhering to treatment are important, since they may affect survival and quality of life of HIV-positive people. PMID- 19133753 TI - A personal consecutive series of surgically treated 51 cases of insular WHO Grade II glioma: advances and limitations. AB - OBJECT: Few experiences of insular surgery have been reported. Moreover, there are no large surgical studies with long-term follow-up specifically dedicated to WHO Grade II gliomas involving the insula. In this paper, the author describes a personal consecutive series of 51 cases in which patients underwent surgery for an insular Grade II glioma. On the basis of the functional and oncological results, advances and limitations of this challenging surgery are discussed. METHODS: Fifty-one patients harboring an insular Grade II glioma (revealed by seizures in 50 cases) underwent surgery. Findings on preoperative neurological examination were normal in 45 patients (88%). All surgeries were conducted under cortico-subcortical stimulation, and in the case of 16 patients while awake. RESULTS: Despite an immediate postoperative worsening in 30 cases (59%), the condition of all but 2 patients (96%) returned to baseline or better. Postoperative MR imaging demonstrated that 77% of resections were total or subtotal. Ten patients underwent a second or third surgery, with no additional deficit. Forty-two patients (82%) are alive with a median follow-up of 4 years. CONCLUSIONS: This is the largest reported experience with insular Grade II glioma surgery. The better knowledge of the insular pathophysiology and the use of intraoperative functional mapping allow the risk of permanent deficit to be minimized (and even enable improvement in quality of life) while increasing the extent of resection and thus the impact on the course of the disease. Therefore, surgical removal must always be considered for insular Grade II glioma. However, this surgery remains challenging, especially within the anterior perforating substance and the posterior part of the (dominant) insula. Additional surgery can be suggested in cases in which the first resection is not complete. PMID- 19133752 TI - Assessment of ultrasound for use in detecting lipoatrophy in HIV-infected patients taking combination antiretroviral therapy. AB - The aim of this study was evaluation of ultrasound (US) as a tool for the assessment of lipoatrophy in a population of HIV-infected patients. We enrolled a convenience sample of 151 HIV-infected Caucasian participants (males, 79%) who were treated for at least 1 year with combination antiretroviral therapy (CART) in Zagreb, Croatia. US measurements of subcutaneous fat thickness were done over the malar, brachial, and crural region. We determined sensitivity and specificity of US as a diagnostic tool for lipoatrophy using receiver-operating curves and concordant patient and clinician assessment as our reference for the presence of lipoatrophy. HIV was acquired through heterosexual contact in 50% of participants and by sex between men in 42%. The mean current CD4 cell count was 503.1 cells=mm3 (standard deviation [SD] = 250.8). Seventy-seven (51%) participants were treated with stavudine and 91 (64%) with a protease inhibitor for at least 6 months. Nineteen (13%)participants had lipoatrophy in at least one anatomic site. Sensitivity of US ranged from 67%-71%, specificity from 65%-71%, positive and negative predictive values ranged from 11%-20% and 96-97%, respectively. US diagnosed lipoatrophy was more frequently found in patients with a history of stavudine treatment and in females. Patients with lipoatrophy had a longer duration of CART than those without lipoatrophy. US is a useful tool in ruling out the presence of clinical lipoatrophy in patients on CART. Using this objective measure of subcutaneous fat may be useful in helping clinicians make decisions about changing therapy. PMID- 19133754 TI - Impact of cytogenetic abnormalities on the management of skull base chordomas. AB - OBJECT: Cytogenetic studies of chordomas are scarce and show multiple changes involving different chromosomes. These abnormalities are implicated in the pathogenesis of chordoma, but the clinical significance of these changes is yet to be determined. In this study, the authors discuss the cytogenetic changes in a large series of skull base chordomas with long-term follow-up and focus on the impact of these changes on the prognosis, progression, and management of the disease. METHODS: The karyotypes of chordomas in 64 patients (36 men and 28 women) were studied in relation to survival and recurrence or progression over a mean follow-up period of 48 +/- 37.5 months. The standard G-banding technique was used for karyotype analysis. Statistical analysis was performed with the Fisher exact test and ORs, and Kaplan-Meier curves were generated for survival and recurrence/progression of disease. RESULTS: Seventy-four percent of de novo chordomas had normal karyotypes and a 3% recurrence rate; there was a 45% recurrence rate in de novo tumors with abnormal karyotypes (p < 0.01). Recurrent tumors were associated with a high incidence of abnormal karyotype (75%). The OR for recurrence in lesions with an abnormal versus a normal karyotype was 12. Aberrations in chromosomes 3, 4, 12, 13, and 14 were associated with frequent recurrence and decreased survival time. Ninety-five percent of cases with progression involved chromosome 3 and/or 13. The median survival time was 4 months when both of these chromosomes had aberrations (p = 0.02). CONCLUSIONS: Chordomas with normal karyotypes were associated with a low rate of recurrence and a long patient survival, and recurrent chordomas were associated with an abnormal karyotype, disease progression, and poor survival. De novo chordomas with normal karyotypes may be amenable to radical resection and adjunctive proton beam therapy. Recurrent and de novo chordomas with abnormal karyotypes were associated with complex cytogenetic abnormalities and a poor prognosis, particularly in the presence of aberrations underlying tumor progression in chromosomes 3, 4, 12, 13, and 14. PMID- 19133755 TI - Incidence of deep venous thrombosis after subarachnoid hemorrhage. AB - OBJECT: The aim of this study was to determine the efficacy of venous ultrasonography in screening for deep venous thrombosis (DVT) after subarachnoid hemorrhage (SAH). A large cohort of patients who had suffered SAH was evaluated with the primary end points of ascertaining the incidence of asymptomatic DVT with venous Doppler ultrasonography screening and of identifying risk factors for the development of DVT and subsequent pulmonary embolism. METHODS: Data from patients with aneurysmal SAH who had been admitted to the neurosurgical intensive care unit (ICU) between December 2002 and October 2006 were retrospectively evaluated. Patients who had undergone surgical or endovascular treatment of an aneurysm following SAH and survived > or = 15 days were included in the study. RESULTS: The overall incidence of DVT among the entire study cohort was 18%. A subgroup analysis identified all patients, with or without symptoms for DVT, who had undergone venous Doppler ultrasonography screening. The incidence of asymptomatic DVT was 24%. Univariate analysis of all patients revealed a significant correlation between the risk of DVT and Hunt and Hess grade (r = 0.38, p < 0.0001), Fisher grade (r = 0.31, p < 0.0001), total hospital stay (r = 0.49, p < 0.0001), and number of days in the ICU (r = 0.48, p < 0.0001). Multivariate analysis demonstrated that the total hospital stay and number of days in the ICU were significant predictors of DVT in all patients (p < 0.0001 and p < 0.0002, respectively). In the subgroup of screened patients, Hunt and Hess grade, total hospital stay, and number of days in the ICU were significant predictors of DVT. Although screened patients were more likely to have DVT (chi(2) = 6.0976, p < 0.02), there was no significant difference in the incidence of DVT or pulmonary embolism between patients who did and those who did not undergo routine lower-extremity Doppler ultrasonography screening. CONCLUSIONS: Routine compressive venous Doppler ultrasonography is an efficient, noninvasive means of identifying DVT as a screening modality in both symptomatic and asymptomatic patients following aneurysmal SAH. The ability to confirm or deny the presence of DVT in this patient population allows one to better identify the indications for chemoprophylaxis. Prophylaxis for venous thromboembolism in neurosurgical patients is common. Emerging literature and anecdotal experience have exposed risks of complications with prophylactic anticoagulation protocols. The identification of patients at high risk-for example, those with asymptomatic DVT-will allow physicians to better assess the role of prophylactic anticoagulation. PMID- 19133756 TI - Nanomechanical motions of cantilevers: direct imaging in real space and time with 4D electron microscopy. AB - The function of many nano- and microscale systems is revealed when they are visualized in both space and time. Here, we report our first observation, using four-dimensional (4D) electron microscopy, of the nanomechanical motions of cantilevers. From the observed oscillations of nanometer displacements as a function of time, for free-standing beams, we are able to measure the frequency of modes of motion and determine Young's elastic modulus and the force and energy stored during the optomechanical expansions. The motion of the cantilever is triggered by molecular charge redistribution as the material, single-crystal organic semiconductor, switches from the equilibrium to the expanded structure. For these material structures, the expansion is colossal, typically reaching the micrometer scale, the modulus is 2 GPa, the force is 600 microN, and the energy is 200 pJ. These values translate to a large optomechanical efficiency (minimum of 1% and up to 10% or more) and a pressure of nearly 1,500 atm. We note that the observables here are real material changes in time, in contrast to those based on changes of optical/contrast intensity or diffraction. PMID- 19133757 TI - Deposition of Cryptosporidium parvum oocysts on natural organic matter surfaces: microscopic evidence for secondary minimum deposition in a radial stagnation point flow cell. AB - A radial stagnation point flow (RSPF) system combined with a microscope was used to determine the deposition kinetics of Cryptosporidium parvum oocysts on quartz surfaces and silica surfaces coated with Suwannee River natural organic matter (SRNOM) in solutions with different ionic strengths. Microscopic evidence of C. parvum oocysts entrapped in the secondary minimum energy well was presented to show that among the entrapped C. parvum oocysts some were washed away by the radial flow and some were able to transfer to deep primary minima and become irreversibly deposited. Experimental data were compared with simulation results obtained by the convective-diffusion equation and Derjaguin-Landau-Verwey Overbeek (DLVO) theory. The experimental results suggested that surface charge heterogeneity led to a higher attachment efficiency at low ionic strength. In addition, the maximum attachment efficiency was less than 1 at high ionic strength due to steric interaction. PMID- 19133758 TI - 3-Alkylpyridinium alkaloids from the Pacific sponge Haliclona sp. AB - The analysis of the polar extracts of the Pacific sponge Haliclona sp. yielded new dimeric (1), trimeric (2), and polymeric 3-alkylpyridinium alkaloids. Their isolation and structural elucidation, based on NMR and MS data, are discussed in detail, along with their cytotoxic activity. PMID- 19133759 TI - Cytotoxic asterosaponins capable of promoting polymerization of tubulin from the starfish Culcita novaeguineae. AB - Four new asterosaponins, novaeguinosides A (1), B (2), C (3), and D (4), were isolated from the bioactive fraction of the starfish Culcita novaeguineae, as active compounds capable of promoting polymerization of tubulin. Their structures were elucidated by extensive spectroscopic studies and chemical evidence. Compounds 1 and 3 are characterized by sulfated side chains not previously found in asterosaponins, and 1 is the first example of a trisulfated asterosaponin. In the side chains of 2 and 4, the 26-carboxylic acid function is found as an amide derivative of taurine, which is a rare feature and first encountered among asterosaponins. All the asterosaponins showed cytotoxicity against two human tumor cell lines. PMID- 19133760 TI - Phase behavior of aqueous dispersions of mixtures of N-palmitoyl ceramide and cholesterol: a lipid system with ceramide-cholesterol crystalline lamellar phases. AB - Ceramides are particularly abundant in the stratum corneum lipid matrix, where they determine its unusual mesostructure, are involved in the lateral segregation of lipid domains in biological cell membranes, and are also known to act as signaling agents in cells. The importance attributed to ceramides in several biological processes has heightened in recent years, demanding a better understanding of their interaction with other membrane components, namely, cholesterol. Structural data concerning pure ceramides in water are relatively scarce, and this is even more the case for mixtures of ceramides with other lipids commonly associated with them in biological systems. We have derived the thermotropic binary phase diagram of mixtures of N-palmitoyl- D-erythro sphingosine, C16:0-ceramide, and cholesterol in excess water, using differential scanning calorimetry and small- and wide-angle X-ray diffraction. These mixtures are self-organized in lamellar mesostructures that, between other particularities, show two ceramide to cholesterol crystalline phases with molar proportions that approach 2:3 and 1:3. The 2:3 phase crystallizes in a tetragonal arrangement with a lamellar repeat distance of 3.50 nm, which indicates an unusual lipid stacking, probably unilamellar. The uncommon mesostructures formed by ceramides with cholesterol should be considered in the rationalization of their singular structural role in biological systems. PMID- 19133761 TI - Effect of interlayer ions on methane hydrate formation in clay sediments. AB - Natural methane hydrates occurring in marine clay sediments exhibit heterogeneous phase behavior with high complexity, particularly in the negatively charged interlayer region. To date, the real clay interlayer effect on natural methane hydrate formation and stability remains still much unanswered, even though a few computer simulation and model studies are reported. We first examined the chemical shift difference of 27Al, 29Si, and 23Na between dry clay and clay containing intercalated methane hydrates (MH) in the interlayer. We also measured the solid-state 13C MAS NMR spectra of MH in Na-montmorillonite (MMT) and Ca montmorillonite (MMT) to reveal abnormal methane popularity established in the course of intercalation and further performed cryo-TEM and XRD analyses to identify the morphology and layered structure of the intercalated methane hydrate. The present findings strongly suggest that the real methane amount contained in natural MH deposits should be reevaluated under consideration of the compositional, structural, and physical characteristics of clay-rich sediments. Furthermore, the intercalated methane hydrate structure should be seriously considered for developing the in situ production technologies of the deep-ocean methane hydrate. PMID- 19133762 TI - Identification of aliphatic and aromatic tertiary N-oxide functionalities in protonated analytes via ion/molecule and dissociation reactions in an FT-ICR mass spectrometer. AB - A mass spectrometric method is presented for the identification of compounds that contain the aliphatic or aromatic N-oxide functional group. This method utilizes gas-phase ion/molecule reactions of tri(dimethylamino)borane (TDMAB), which rapidly derivatizes protonated aliphatic and aromatic tertiary N-oxides, amides, and some amines via loss of dimethylamine in a Fourier transform ion cyclotron resonance mass spectrometer. The mechanism involves proton transfer from the protonated analyte to the borane, followed by addition of the analyte to the boron center and elimination of dimethylamine. The derivatized analytes are readily identified on the basis of their m/z value which is 98 Th (thomson) greater than that of the protonated analyte, and the characteristic boron isotope patterns. SORI-CAD of the product ions (adduct-(CH3)2NH) yields different fragment ions for aliphatic tertiary N-oxides, aromatic tertiary N-oxides, amides, and pyridines. Therefore, these analytes can be identified based on their characteristic fragment ions. This method was tested by examining two drug samples, Olanzapine and Olanzapine-4' N-oxide. PMID- 19133763 TI - Effect of headgroup chirality in nanoassemblies. Part 1. Self-assembly of D isoascorbic acid derivatives in water. AB - L-(+)-Ascorbic acid and D-(-)-isoascorbic acid are epimers, with an opposite configuration at the C5 stereogenic chiral center. Single-chained surfactants that carry a L-ascorbic or d-isoascorbic acid residue as hydrophilic headgroup and an alkanoate tail as hydrophobic chain were synthesized. We refer to these as L-ASCn and D-ASCn, with n=8, 10, or 12. The role of the headgroup configuration in determining the nature of both the pure compounds and their nano assembly in aqueous dispersions were studied. Surface tension, infrared spectroscopy, differential scanning calorimetry, conductivity, small-angle X-ray scattering, and wide-angle X-ray diffractometry were used to characterize surfactant properties as a function of temperature and concentration. The L and D headgroup forms have significantly different hydration. This greatly affects the structure and stability of the aggregates in the micellar and coagel states. PMID- 19133764 TI - Femtosecond laser-nanostructured substrates for surface-enhanced Raman scattering. AB - We present a new type of surface-enhanced Raman scattering (SERS) substrate that exhibits extremely large and uniform cross-section enhancements over a macroscopic (greater than 25 mm2) area. The substrates are fabricated using a femtosecond laser nanostructuring process, followed by thermal deposition of silver. SERS signals from adsorbed molecules show a spatially uniform enhancement factor of approximately 10(7). Spectroscopic characterization of these substrates suggests their potential for use in few or single-molecule Raman spectroscopy. PMID- 19133765 TI - Halogen bonded supramolecular assemblies of [Ru(bipy)(CN)4]2- anions and N-methyl halopyridinium cations in the solid state and in solution. AB - The interactions between the [Ru(bipy)(CN)(4)](2-) anion and N-methyl halopyridinium cations have been examined in both the solid state and in solution. In the solid state, crystal structures of [Ru(bipy)(CN)(4)](2-) salts containing iodinated cations (N-methyl-3-iodopyridinium and N-methyl-3,5 diiodopyridinium) show clear C-I...NC(Ru) halogen bonds between the externally directed cyanide lone pairs of the anion and the iodine atoms of the cation which dominates the structures. In contrast the analogous brominated cations (N-methyl 3-bromopyridinium and N-methyl-3,5-dibromopyridinium) do not exhibit C Br...NC(Ru) interactions in the solid state, with the cyanide groups instead involved in hydrogen bonding, principally to lattice water molecules. The charge assisted C-I...NC(Ru) interactions are therefore clearly of value as synthons in crystal engineering applications. In CH(2)Cl(2) solution, spectroscopic titrations between [Ru(4,4'-(t)Bu(2)-bipy)(CN)(4)](2-) and both N-methyl-3 iodopyridinium and N-methyl-3-bromopyridinium cations show clear evidence for formation of distinct 1:1, 3:2, and then 2:1 cation/anion adducts with high association constants (>10(7) M(-1) for the first 1:1 association constant). However the presence of identical results using the non-halogenated cation N methyl-pyridinium indicates that this strong cation/anion association in CH(2)Cl(2) is dominated by electrostatic effects: either C-H...NC(Ru) hydrogen bonds or C-X...NC(Ru) halogen bonds could be involved in the ion pairs but it is the charge-assistance that makes the association strong. This is confirmed by a titration between [Ru(4,4'-(t)Bu(2)-bipy)(CN)(4)](2-) and the neutral halogen bond acceptor C(6)F(5)I for which the first association constant is very low (ca. 6 M(-1)). The formation of adducts between [Ru(4,4'-(t)Bu(2)-bipy)(CN)(4)](2-) and the various N-methyl-pyridinium cations in solution results in a clear blue shift of the (1)MLCT absorption maxima associated with the Ru(II) unit, a characteristic consequence of interaction of the cyanide lone pairs with a Lewis acidic site on the cation. The (3)MLCT luminescence from the [Ru(4,4'-(t)Bu(2) bipy)(CN)(4)](2-) center, however, does not show the usual associated increase in intensity associated with this blue shift in the (1)MLCT absorptions, most likely because of electron-transfer quenching by the N-methyl-pyridinium cations in the assemblies. PMID- 19133766 TI - Nonaromatic core-shell structure of nanodiamond from solid-state NMR spectroscopy. AB - The structure of synthetic nanodiamond has been characterized by (13)C nuclear magnetic resonance (NMR) spectral editing combined with measurements of long range (1)H-(13)C dipolar couplings and (13)C relaxation times. The surface layer of these approximately 4.8-nm diameter carbon particles consists mostly of sp(3) hybridized C that is protonated or bonded to OH groups, while sp(2)-hybridized carbon makes up less than 1% of the material. The surface protons surprisingly resonate at 3.8 ppm, but their direct bonding to carbon is proved by fast dipolar dephasing under homonuclear decoupling. Long-range (1)H-(13)C distance measurements, based on (13)C{(1)H} dipolar dephasing by surface protons, show that seven carbon layers, in a shell of 0.63 nm thickness that contains approximately 60% of all carbons, predominantly resonate more than +8 ppm from the 37-ppm peak of bulk diamond (i.e., within the 45-80 ppm range). Nitrogen detected in (15)N NMR spectra is mostly not protonated and can account for some of the high-frequency shift of carbon. The location of unpaired electrons (approximately 40 unpaired electrons per particle) was studied in detail, based on their strongly distance-dependent effects on T(1,C) relaxation. The slower relaxation of the surface carbons, selected by spectral editing, showed that the unpaired electrons are not dangling bonds at the surface. This was confirmed by detailed simulations, which indicated that the unpaired electrons are mostly located in the disordered shell, at distances between 0.4 and 1 nm from the surface. On the basis of these results, a nonaromatic core-shell structural model of nanodiamond particles has been proposed. PMID- 19133767 TI - Design, selection, and characterization of thioflavin-based intercalation compounds with metal chelating properties for application in Alzheimer's disease. AB - Metal chelation is considered a rational therapeutic approach for interdicting Alzheimer's amyloid pathogenesis. At present, enhancing the targeting and efficacy of metal-ion chelating agents through ligand design is a main strategy in the development of the next generation of metal chelators. Inspired by the traditional dye Thioflavin-T, we have designed new multifunctional molecules that contain both amyloid binding and metal chelating properties. In silico techniques have enabled us to identify commercial compounds that enclose the designed molecular framework (M1), include potential antioxidant properties, facilitate the formation of iodine-labeled derivatives, and can be permeable through the blood-brain barrier. Iodination reactions of the selected compounds, 2-(2 hydroxyphenyl)benzoxazole (HBX), 2-(2-hydroxyphenyl)benzothiazole (HBT), and 2-(2 aminophenyl)-1H-benzimidazole (BM), have led to the corresponding iodinated derivatives HBXI, HBTI, and BMI, which have been characterized by X-ray diffraction. The chelating properties of the latter compounds toward Cu(II) and Zn(II) have been examined in the solid phase and in solution. The acidity constants of HBXI, HBTI, and BMI and the formation constants of the corresponding ML and ML2 complexes [M = Cu(II), Zn(II)] have been determined by UV-vis pH titrations. The calculated values for the overall formation constants for the ML2 complexes indicate the suitability of the HBXI, HBTI, and BMI ligands for sequestering Cu(II) and Zn(II) metal ions present in freshly prepared solutions of beta-amyloid (Abeta) peptide. This was confirmed by Abeta aggregation studies showing that these compounds are able to arrest the metal-promoted increase in amyloid fibril buildup. The fluorescence features of HBX, HBT, BM, and the corresponding iodinated derivatives, together with fluorescence microscopy studies on two types of pregrown fibrils, have shown that HBX and HBT compounds could behave as potential markers for the presence of amyloid fibrils, whereas HBXI and HBTI may be especially suitable for radioisotopic detection of Abeta deposits. Taken together, the results reported in this work show the potential of new multifunctional thioflavin-based chelating agents as Alzheimer's disease therapeutics. PMID- 19133768 TI - Fluorous polymeric membranes for ionophore-based ion-selective potentiometry: how inert is Teflon AF? AB - Fluorous media are the least polar and polarizable condensed phases known. Their use as membrane materials considerably increases the selectivity and robustness of ion-selective electrodes (ISEs). In this research, a fluorous amorphous perfluoropolymer was used for the first time as a matrix for an ISE membrane. Electrodes for pH measurements with membranes composed of poly[4,5-difluoro-2,2 bis(trifluoromethyl)-1,3-dioxole]-co-poly(tetrafluoroethylene) (87% dioxole monomer content; known as Teflon AF2400) as polymer matrix, a linear perfluorooligoether as plasticizer, sodium tetrakis[3,5 bis(perfluorohexyl)phenyl]borate providing for ionic sites, and bis[(perfluorooctyl)propyl]-2,2,2-trifluoroethylamine as H+ ionophore were investigated. All electrodes had excellent potentiometric selectivities, showed Nernstian responses to H+ over a wide pH range, exhibited enhanced mechanical stability, and maintained their selectivity over at least 4 weeks. For membranes of low ionophore concentration, the polymer affected the sensor selectivity noticeably at polymer concentrations exceeding 15%. Also, the membrane resistance increased quite strongly at high polymer concentrations, which cannot be explained by the Mackie-Meares obstruction model. The selectivities and resistances depend on the polymer concentration because of a functional group associated with Teflon AF2400, with a concentration of one functional group per 854 monomer units of the polymer. In the fluorous environment of these membranes, this functional group binds to Na+, K+, Ca2+, and the unprotonated ionophore with binding constants of 10(3.5), 10(1.8), 10(6.8), and 10(4.4) M(-1), respectively. Potentiometric and spectroscopic evidence indicates that these functional groups are COOH groups formed by the hydrolysis of carboxylic acid fluoride (COF) groups originally present in Teflon AF2400. The use of higher ionophore concentrations removes the undesirable effect of these COOH groups almost completely. Alternatively, the C(=O)F groups can be eliminated chemically, or they can be used to readily introduce new functionalities. PMID- 19133769 TI - Stability of superoxide ion in imidazolium cation-based room-temperature ionic liquids. AB - The stability of superoxide ion (O(2)(*-)) generated chemically by dissolving KO(2) in dried dimethyl sulfoxide solutions containing imidazolium cation [e.g., 1-ethyl-3-methylimidazolium (EMI(+)) and 1-n-butyl-2,3-dimethylimidazolium (BMMI(+))] based ionic liquids (ILs) was investigated with UV-visible spectroscopic, NMR, and voltammetric techniques and an ab initio molecular orbital calculation. UV-visible spectroscopic and cyclic voltammetric measurements reveal that the O(2)(*-) species reacts with BMMI(+) and EMI(+) cations of ILs to form hydrogen peroxide. The pseudo first order rate constant for the reaction of BMMI(+) and O(2)(*-) species was found to be about 2.5 x 10( 3) s(-1). With a molecular orbital calculation, the O(2)(*-) species is understood to attack the 2-position (C-2) of the imidazolium ring (i.e., BMMI(+)) to form an ion pair complex in which one oxygen atom is bounded to C-2 and the other to the hydrogen atom of -CH(3) group attached to C-2. Eventually, the ion pair complex of BMMI(+) cation and O(2)(*-) species undergoes a ring opening reaction as evidenced with (1)H NMR measurement. PMID- 19133770 TI - Scope of the Suzuki-Miyaura cross-coupling reaction of potassium trifluoroboratoketohomoenolates. AB - Potassium trifluoroboratoketohomoenolates were prepared in good yields from either the corresponding alpha,beta-unsaturated compounds or methyl ketones. These organoboron reagents were effectively cross-coupled with various aryl and heteroaryl chlorides. PMID- 19133771 TI - Photoactivable antibody binding protein: site-selective and covalent coupling of antibody. AB - Here we report new photoactivable antibody binding proteins, which site selectively capture antibodies and form covalent conjugates with captured antibodies upon irradiation. The proteins allow the site-selective tagging and/or immobilization of antibodies with a highly preferred orientation and omit the need for prior antibody modifications. The minimal Fc-binding domain of protein G, a widely used antibody binding protein, was genetically and chemically engineered to contain a site-specific photo cross-linker, benzophenone. In addition, the domain was further mutated to have an enhanced Fc-targeting ability. This small engineered protein was successfully cross-linked only to the Fc region of the antibody without any nonspecific reactivity. SPR analysis indicated that antibodies can be site-selectively biotinylated through the present photoactivable protein. Furthermore, the system enabled light-induced covalent immobilization of antibodies directly on various solid surfaces, such as those of glass slides, gold chips, and small particles. Antibody coupling via photoactivable antibody binding proteins overcomes several limitations of conventional approaches, such as random chemical reactions or reversible protein binding, and offers a versatile tool for the field of immunosensors. PMID- 19133772 TI - Efficient asymmetric transfer hydrogenation of ketones catalyzed by an iron complex containing a P-N-N-P tetradentate ligand formed by template synthesis. AB - A distorted iron complex formed by template synthesis is the basis of a very efficient and economical catalyst system for the asymmetric transfer hydrogenation of ketones for the production of valuable enantioenriched alcohols. PMID- 19133773 TI - A role of the heme-7-propionate side chain in cytochrome P450cam as a gate for regulating the access of water molecules to the substrate-binding site. AB - Cytochrome P450cam is a heme-containing enzyme which catalyzes hydroxylation of d camphor. The heme is bound in the heme pocket via noncovalent interactions, where two heme-propionate side chains interact with Arg, His, and/or Asp residues. To understand the role of the heme-7-propionate side chain, we prepared reconstituted P450cam with an artificial one-legged heme which has a methyl group at the position of the 7-propionate. Removal of 7-propionate dramatically decreases the d-camphor affinity by 3 orders of magnitude relative to that of the wild-type enzyme, and spectroscopic data indicate that 74% of the ferric P450cam exhibits a low-spin state owing to water molecule occupancy in the substrate binding site under the normal assay conditions. Thus, the monooxygenase activity of the reconstituted protein is remarkably low due to the decrease in the rate of the first electron transfer from reduced putidaredoxin, whereas 87% of oxidized NADH was utilized to produce 5-hydroxy-d-camphor without any significant uncoupling reactions. X-ray structural analysis of the reconstituted enzyme reveals a novel water array extending from the substrate-binding site to bulk solvent through the position occupied by 7-propionate. This water array appears without causing any major changes in the protein structure with the notable exception of conformational changes occurring at Asp297 and Gln322 residues. We propose that the 7-propionate forms a barrier against entry of bulk water molecules and therefore in combination with Asp297, Arg299, and Gln322 plays an essential role in the process of elimination of the substrate-binding site water cluster which occurs upon d-camphor binding. PMID- 19133774 TI - An acyclic dimer of cyclodiphosphazane {(t)BuHN((t)BuNP)2OCH2}2 containing alkoxo and amido functionalities: synthesis, derivatization, Bi- (Pd(II), Rh(I)), and tetranuclear (Pd(II), Au(I), Rh(I)Au(I)) transition metal complexes. AB - Acyclic bis(cyclodiphosphazane) [(t)BuHN((t)BuNP)(2)OCH(2)](2) (2) containing alkoxo and amido functionalities was synthesized by reacting cis [(t)BuHN((t)BuNP)(2)Cl] (1) with ethylene glycol and upon further treatment with 4 equiv of elemental sulfur or selenium affords the corresponding tetrachalcogenides [(t)BuHN((t)BuNPE)(2)OCH(2)](2) (3, E = S; 4, E = Se) in quantitative yield. The reaction of 2 with 2 equiv of elemental sulfur results in the oxidation of only the amido-phosphorus atoms to form the di(sulfide) derivative [(t)BuHN(S)P(mu-(t)BuN)POCH(2)](2) (5). The reaction of 2 with 2 equiv of [PdCl(eta(3)-C(3)H(5))](2) afforded the tetrametallic complex [{Pd(eta(3) allyl)Cl}(4){(t)BuHN((t)BuNP)(2)OCH(2)}(2)] (7) containing four independent [PdCl(2)(eta(3)-C(3)H(5))] moieties each coordinated to a phosphorus atom. In contrast, the reaction between [Pd(PEt(3))Cl(2)](2) and 2 leads to the addition of two metal atoms with the formation of [{Pd(PEt(3))Cl}(2){(t)BuHN((t)BuNP)(2)OCH(2)}(2)] (8), in which alternating phosphorus atoms of the bis(cyclodiphosphazane) are coordinated to palladium. Interestingly, the palladium(II) atom coordinated to the phosphorus atom bearing the t-butylamino substituent adopts a trans geometry, whereas that coordinated to the phosphorus atom connected to the -OCH(2)CH(2)O- linker prefers a cis conformation. The reaction between 2 and [Rh(COD)Cl](2) produces exclusively the bimetallic complex [{Rh(COD)Cl}(2){(t)BuHN((t)BuNP)(2)OCH(2)}(2)] (6) irrespective of the stoichiometry of the reactants and the reaction conditions. In complex 6, only the alkoxo-phosphorus atoms are coordinated. However, complex 6 upon treatment with AuCl(SMe(2)) in a 1:2 ratio gives the heterometallic tetrasubstituted complex [{Rh(COD)Cl}(2)(AuCl)(2){(t)BuHN((t)BuNP)(2)OCH(2)}(2)] (10). The reaction between 4 equiv of AuCl(SMe(2)) and 2 resulted in the formation of a tetrametallic gold complex [(AuCl)(4){(t)BuHN((t)BuNP)(2)OCH(2)}(2)] (9). The crystal structures of 2-4 and 7-10 are reported. PMID- 19133775 TI - The translocator protein (18 kDa): central nervous system disease and drug design. PMID- 19133776 TI - Guanidine and 2-aminoimidazoline aromatic derivatives as alpha2-adrenoceptor ligands: searching for structure-activity relationships. AB - In this paper, we report the synthesis of three new 2-aminoimidazoline (compounds 4b, 5b, and 6b) and three new guanidine derivatives (compounds 7b, 8b, and 9b) as potential alpha(2)-adrenoceptor antagonists for the treatment of depression. Their pharmacological profile was evaluated in vitro in human brain tissue and compared to the potential antidepressant 1 and the agonists 2 and 3. All new substrates were evaluated by in vitro functional [(35)S]GTPgammaS binding assays in human prefrontal cortex to determine their agonistic or antagonistic activity. Compound 8b was found to be an antagonist in vitro and was subjected to in vivo microdialysis experiments in rats. Moreover, a new synthesis of the precursor amines for compounds 4b-9b is presented. PMID- 19133777 TI - Effect of fluorination on the pharmacological profile of 11beta isomers of fulvestrant in breast carcinoma cells. AB - We describe the synthesis of an 11beta isomer 3 of the steroidal antiestrogen fulvestrant 2. Partial fluorination of the 11beta side chain in 3 leads to 4, which still shows strong antiproliferative activity on MCF-7 cells. However, unlike 2 and 3, compound 4 fails to down-regulate estrogen receptor alpha (ERalpha). This result suggests that ERalpha down-regulation is not a sine qua non condition for the antitumor activity of steroidal antiestrogens. PMID- 19133778 TI - Rhazinilam and quebrachamine derivatives from Yunnan Kopsia arborea. AB - Three new rhazinilam-derived alkaloids, kopsiyunnanines C1, C2, and C3, and a new quebrachamine-type alkaloid, kopsiyunnanine D, which possess an unusual methoxymethyl or ethoxymethyl function, were isolated from the aerial parts of Yunnan Kopsia arborea. This is the first report of the presence of these functions in natural alkaloids. The structures and absolute configurations of the alkaloids were determined by spectroscopic methods and confirmed by semisynthesis. PMID- 19133779 TI - Antiprotealide is a natural product. AB - Large-scale fermentation of the marine actinomycete Salinispora tropica for production of salinosporamide A (NPI-0052; 1) clinical trials materials provided crude extracts containing minor secondary metabolites, including salinosporamide B (2) and a new congener, 3. Spectroscopic characterization revealed that 3 is identical to antiprotealide, a molecular hybrid of 20S proteasome inhibitors 1 and omuralide (4) not previously described as a natural product. Analysis of crude extracts from shake flask cultures of three wild-type S. tropica strains confirmed the production of antiprotealide at 1.1, 0.8, and 3.0 mg/L. Thus, antiprotealide is a natural product metabolite of S. tropica. PMID- 19133780 TI - Cytotoxic triterpenoid alkaloids from Buxus microphylla. AB - Five new triterpenoid alkaloids, buxmicrophyllines E-I (1-5), and six known ones (6-11) were isolated from the leaves and stems of Buxus microphylla. The structures of compounds 1-5 were elucidated by NMR and MS spectroscopic analysis, and the relative stereochemistry of 5 was determined by single-crystal X-ray crystallography. Compounds 3 and 9 were cytotoxic against HepG2 cells, with IC(50) values of 0.89 and 0.78 microM, and compounds 2, 3, 7, 8, and 9 were cytotoxic against K562 cells, with IC(50) values of 2.95, 4.44, 1.70, 5.61, and 0.37 microM, respectively. PMID- 19133781 TI - Host-guest complexes with protein-ligand-like affinities: computational analysis and design. AB - It has recently been discovered that guests combining a nonpolar core with cationic substituents bind cucurbit[7]uril (CB[7]) in water with ultrahigh affinities. The present study uses the Mining Minima algorithm to study the physics of these extraordinary associations and to computationally test a new series of CB[7] ligands designed to bind with similarly high affinity. The calculations reproduce key experimental observations regarding the affinities of ferrocene-based guests with CB[7] and beta-cyclodextrin and provide a coherent view of the roles of electrostatics and configurational entropy as determinants of affinity in these systems. The newly designed series of compounds is based on a bicyclo[2.2.2]octane core, which is similar in size and polarity to the ferrocene core of the existing series. Mining Minima predicts that these new compounds will, like the ferrocenes, bind CB[7] with extremely high affinities. PMID- 19133782 TI - X-ray structure of native scorpion toxin BmBKTx1 by racemic protein crystallography using direct methods. AB - Racemic protein crystallography, enabled by total chemical synthesis, has allowed us to determine the X-ray structure of native scorpion toxin BmBKTx1; direct methods were used for phase determination. This is the first example of a protein racemate that crystallized in space group I41/a. PMID- 19133783 TI - Iodomesitylene-catalyzed oxidative cleavage of carbon-carbon double and triple bonds using m-chloroperbenzoic acid as a terminal oxidant. AB - Transition metal-catalyzed oxidative cleavage of carbon-carbon multiple bonds has emerged as a powerful tool in organic synthesis. High-valent oxometals, mostly of Ru, Os, Mn, Mo, W, and Re, were used catalytically as reactive oxygen transfer agents to the multiple bonds. Reported here for the first time are the organocatalytic versions of the oxidative cleavage reactions. Our method involves use of iodomesitylene as an effective organocatalyst, which generates an active aryl(hydroxy)-lambda(3)-iodane 5 in situ, and m-chloroperbenzoic acid (m-CPBA) as a terminal oxidant under metal-free conditions. Cyclic and acyclic olefins as well as aliphatic and aromatic alkynes were smoothly cleaved to carboxylic acids under the organocatalytic conditions. PMID- 19133785 TI - Digitally-multiplexed nanoelectrospray ionization atmospheric pressure drift tube ion mobility spectrometry. AB - One of the shortcomings of atmospheric pressure drift tube ion mobility spectrometry (DTIMS) is its intrinsically low duty cycle (approximately 0.04-1%) caused by the rapid pulsing of the ion gate (25-400 micros) followed by a comparatively long drift time (25-100 ms), which translates into a loss of sensitivity. Multiplexing approaches via Hadamard and Fourier-type gating techniques have been reported for increasing the sensitivity of DTIMS. Here, we report an extended multiplexing approach which encompasses arbitrary binary ion injection waveforms with variable duty cycles ranging from 0.5 to 50%. In this approach, ion mobility spectra can be collected using conventional signal averaging, arbitrary, standard Hadamard and/or "extended" Hadamard operation modes. Initial results indicate signal-to-noise gains ranging from 2-7-fold for both arbitrary and "extended" Hadamard sequences. Standard Hadamard transform IMS provided increased sensitivity, with gains ranging from 9-12-fold, however, mobility spectra suffered from defects that appeared as false peaks, which were reduced or eliminated when using arbitrary or "extended" Hadamard waveforms for multiplexing. Digital multiplexing enables variation of the duty cycle in a continuous manner, minimizing the contribution of imperfect modulation on spectral defects without the need for complex spectral correction methods. By reducing the frequency of gating events employed in the variable duty cycle sequences, the contributions of factors such as ion depletion prior to gating, interaction of successively injected ion packets, and the cumulative effect of imperfect gating events were mitigated. PMID- 19133784 TI - Prion disease diagnosis by proteomic profiling. AB - Definitive prion disease diagnosis is currently limited to postmortem assay for the presence of the disease-associated proteinase K-resistant prion protein. Using cerebrospinal fluid (CSF) from prion-infected hamsters, matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS), and support vector machines (SVM), we have identified peptide profiles characteristic of disease state. Using 10-fold leave-one-out cross-validation, we report a predictive accuracy of 72% with a true positive rate of 73% and a false positive rate of 27% demonstrating the suitability of using proteomic profiling and CSF for the development of multiple marker diagnostics of prion disease. PMID- 19133786 TI - Kinetic studies on the thermal cis-to-trans isomerization of 1-phenyltriazenes derived from cyclic amines. AB - Rate constants for thermal cis-to-trans isomerization of N-(phenylazo) substituted nitrogen ring heterocyles were determined as a function of phenyl ring substitution, cyclic amine ring size, and organic solvents. Observed first order rate constants are found to increase with increasing electron-withdrawing character of the para substituent, with larger amine rings, and with increasing solvent polarity. Overall, trends observed are consistent with geometrical isomerization taking place through rotation around the nitrogen-nitrogen double bond via a polarized transition state. PMID- 19133788 TI - Mechanism for activation of molecular oxygen by cis- and trans (pyridine)2Pd(OAc)H: Pd(0) versus direct insertion. AB - We use quantum mechanics to elucidate the mechanism for the reaction of molecular oxygen with palladium-hydride complexes, (pyridine)(2)-Pd(II)(H)OAc, in toluene, focusing specifically on the direct insertion pathway of dioxygen into the Pd-H bond and pathways proceeding through a Pd(0) intermediate for both cis and trans starting configurations as well as with the assistance of an extra HOAc molecule We report the potential energy surfaces and structures for each of these pathways. This is the first examination of these two mechanisms for both cis and trans isomers of a system employing two monodentate ligands. It is the first case of acid-assisted reductive elimination from a square planar Pd-H. The calculated mechanisms indicate that cis/trans isomerization is feasible and demonstrates that the Pd(0) pathway is favored for both cis (DeltaDeltaH(double dagger) = 2.2 kcal/mol, DeltaDeltaG(double dagger) = 9.3 kcal/mol) and trans cases (HOAc assisted; DeltaDeltaH(double dagger) = -2.6 kcal/mol, DeltaDeltaG(double dagger) = 5.8 kcal/mol) demonstrating that the presence of two monodentate ligands changes the mechanism from that of the bidentate case. PMID- 19133787 TI - Orientation and peptide-lipid interactions of alamethicin incorporated in phospholipid membranes: polarized infrared and spin-label EPR spectroscopy. AB - Alamethicin is a 20-residue peptaibiotic that induces voltage-dependent ion channels in lipid membranes. The mode by which alamethicin inserts into membranes was investigated using measurements of peptide-lipid interactions by spin-label electron paramagnetic resonance (EPR) and of peptide orientation by polarized infrared (IR) spectroscopy. In fluid membranes, spin-labeled stearic acid shows no evidence of a specific motionally restricted population of lipid chains, such as that found at the intramembranous surface of integral membrane proteins or oligomeric assemblies of transmembrane alpha-helices. In agreement with recent results from TOAC-substituted alamethicin analogues, native alamethicin is predominantly monomeric in fluid lipid membranes and presents an intramembrane surface that integrates well with the lipid chains but is insufficiently extensive to induce specific motional restriction. Channel formation takes place by transient association of transmembrane monomers. In aligned fluid membranes, alamethicin exhibits a large tilt in short chain-length lipids that decreases first rapidly with increasing chain-length and then more gradually for the lipids with longer chains. This macroscopically low order contrasts with the high local order, relative to the local membrane normal, that is found by EPR for alamethicins spin-labeled with TOAC. The macroscopic behavior is consistent with predictions for the chain-length dependence of elastic bending fluctuations of the membrane surface, which was invoked recently to explain the spontaneous insertion of beta-barrel proteins in short-chain lipid membranes. PMID- 19133789 TI - Multifunctional polypyrrole/strontium hexaferrite composite microspheres: preparation, characterization, and properties. AB - Polypyrrole (PPY)/SrFe12O19 composites with tunable electrical and magnetic properties were synthesized by in situ polymerization of pyrrole in the presence of SrFe12O19 particles. The structure of PPY/SrFe12O19 composites was characterized by means of Fourier transform infrared spectroscopy and X-ray diffraction. Transmission electron microscope and scanning electron microscope images illustrated that the spherical composites consisted of SrFe12O19 hexagonal plates sheathed by PPY. In the electromagnetic measurments, it was found that the ac conductivity of SrFe12O19 particles increased while the saturation magnetization, remanent magnetization, and coercivity decreased after PPY coating; moreover, the desired electrical and magnetic properties of PPY/SrFe12O19 composites can be modulated simply by controlling the contents of SrFe12O19 particles. A possible mechanism was also proposed to interpret the formation of the PPY/SrFe12O19 composites. PMID- 19133790 TI - Improving the stability and sensing of electrochemical biosensors by employing trithiol-anchoring groups in a six-carbon self-assembled monolayer. AB - Alkane thiol self-assembled monolayers (SAMs) have seen widespread utility in the fabrication of electrochemical biosensors. Their utility, however, reflects a potentially significant compromise. While shorter SAMs support efficient electron transfer, they pack poorly and are thus relatively unstable. Longer SAMs are more stable but suffer from less efficient electron transfer, thus degrading sensor performance. Here we use the electrochemical DNA (E-DNA) sensor platform to compare the signaling and stability of biosensors fabricated using a short, six carbon monothiol with those employing either of two commercially available trihexylthiol anchors (a flexible Letsinger type and a rigid adamantane type). We find that all three anchors support efficient electron transfer and E-DNA signaling, with the gain, specificity, and selectivity of all three being effectively indistinguishable. The stabilities of the three anchors, however, vary significantly. Sensors anchored with the flexible trithiol exhibit enhanced stability, retaining 75% of their original signal and maintaining excellent signaling properties after 50 days storage in buffer. Likewise these sensors exhibit excellent temperature stability and robustness to electrochemical interrogation. The stability of sensors fabricated using the rigid trithiol anchor, by comparison, are similar to those of the monothiol, with both exhibiting significant (>60%) loss of signal upon wet storage or thermocycling. Employing a flexible trithiol anchor in the fabrication of SAM-based electrochemical biosensors may provide a means of improving sensor robustness without sacrificing electron transfer efficiency or otherwise impeding sensor performance. PMID- 19133791 TI - Molecular gradients of bioinertness reveal a mechanistic difference between mammalian cell adhesion and bacterial biofilm formation. AB - Chemical gradients play an important role in guiding the activities of both eukaryotic and prokaryotic cells. Here, we used molecularly well-defined chemical gradients formed by self-assembled monolayers (SAMs) on gold films to reveal that mammalian cell adhesion and bacterial biofilm formation respond differently to a gradient of surface chemistry that resists cell attachment. Gradient self assembled monolayers (SAMs) consisting of two mixed alkanethiols were fabricated by differential exposure of the gold film to one alkanethiol, followed by soaking in another alkanethiol solution. A gradient in bioinertness that resisted cell attachment was created on SAMs from a gradient in the surface density of HS(CH2)11(OCH2CH2)3OH, backfilled with either HS(CH2)11OH or HS(CH2)11CH3. Measurements of the amounts of mammalian cells and bacterial biofilms on these gradient surfaces reveal that, for mammalian cells, a critical density of adhesion ligands from absorbed proteins on surfaces exists for supporting maximum adhesion and proliferation, whereas for the bacterium Escherichia coli , the amount of biofilm formed on surfaces increased linearly with the surface density of adhesive groups (methyl or hydroxyl groups) in different media. These results are consistent with mammalian cell adhesion requiring an anchorage via specific molecular recognitions and suggest that biofilms can form by immobilization of bacteria via nonspecific interaction between bacteria and surfaces. PMID- 19133792 TI - Visualization of enantiomers in the liquid-crystalline phase of a fragmented DNA solution. AB - The use of the liquid-crystalline phase of fragmented DNA solution for enantiomeric differentiation by NMR is reported. The lyotropic cholesteric liquid crystal system formed, orients in a magnetic field and is able to discriminate water soluble enantiomeric mixtures in a simple 2D J-resolved NMR experiment. PMID- 19133794 TI - Perfluorinated polycyclic aromatic hydrocarbons: anthracene, phenanthrene, pyrene, tetracene, chrysene, and triphenylene. AB - The properties of perfluoroanthracene (1-C(14)F(10)), perfluorophenanthrene (2 C(14)F(10)), perfluoropyrene (C(16)F(10)), perfluorotetracene (1-C(18)F(12)), perfluorochrysene (2-C(18)F(12)), and perfluorotriphenylene (3-C(18)F(12)) and their radical anions have been studied using density functional theory (DFT). Three measures of neutral-anion energy separations reported in this work are the adiabatic electron affinity (EA(ad)), the vertical electron affinity (EA(vert)), and the vertical detachment energy (VDE). The vibrational frequencies of these perfluoro PAHs and their radical anions are also examined. The predicted adiabatic electron affinities (DZP++ B3LYP) are: 1.84 eV, 1-C(14)F(10); 1.41 eV, 2-C(14)F(10); 1.72 eV, C(16)F(10); 2.39 eV, 1-C(18)F(12); 1.83 eV (C(i) symmetry) and 1.88 eV (C(2) symmetry), 2-C(18)F(12); and 1.69 eV, 3-C(18)F(12). The perfluorotetracene is clearly the most effective electron captor. Perfluorophenanthrene, perfluoropyrene, perfluorochrysene, and perfluorotriphenylene, as well as their radical anions deviate from planarity. For example, the nonplanar perfluorochrysene structures are predicted to lie 7-13 kcal/mol below the pertinent C(2h) stationary points. PMID- 19133795 TI - Systematic first principles parameterization of force fields for metal-organic frameworks using a genetic algorithm approach. AB - A systematic strategy is proposed to derive the necessary force field parameters directly from first principles calculations of nonperiodic model systems to reproduce both the structure and curvature of the reference potential energy surface. The parameters are determined using a genetic algorithm combined with a novel fitness criterion based on a representation of structure and curvature in a set of redundant internal coordinates. Due to the efficiency of this approach it is possible to abandon the need for transferability of the parameters. The method is targeted for the application on metal-organic frameworks (MOFs), where parameters for molecular mechanics force fields are often not available, because of the wide range of possible inorganic fragments involved. The scheme is illustrated for Zn4O-based IRMOF materials on the example of MOF-5. In a "building block" approach parameters are derived for the two model systems basic zinc formate (Zn4O(O2CH)6), and dilithium terephthalate with reference data obtained from density functional theory. The resulting potential gives excellent agreement with the structure, vibrational frequencies, thermal behavior and elastic constants of the periodic MOF-5. PMID- 19133796 TI - Neuraminidase inhibitor resistance after oseltamivir treatment of acute influenza A and B in children. AB - BACKGROUND: Oseltamivir, a specific influenza neuraminidase inhibitor, is an effective treatment for seasonal influenza. Emergence of drug-resistant influenza viruses after treatment has been reported, particularly in children in Japan, where the dosing schedule is different from that used throughout the rest of the world. We investigated the emergence of drug-resistant infection in children treated with a tiered weight-based dosing regimen. METHODS: We analyzed sequential clinical nasopharyngeal samples, obtained before and after tiered weight-based oseltamivir therapy, from children with acute influenza during 2005 2007. We isolated viruses, tested for drug resistance with use of a fluorescence based neuraminidase inhibition assay, performed neuraminidase gene sequencing, and determined quantitative viral loads. RESULTS: Sixty-four children (34 with influenza A subtype H3N2, 11 with influenza A subtype H1N1, and 19 with influenza B virus) aged 1-12 years (median age, 3 years, 1 month) were enrolled. By days 4 7 after initiation of treatment, of 64 samples tested, 47 (73.4%) and 26 (40.6%) had virus detectable by reverse-transcriptase polymerase chain reaction and culture, respectively. By days 8-12 after initiation of treatment, of 53 samples tested, 18 (33.9%) and 1 (1.8%) had virus detectable by reverse-transcriptase polymerase chain reaction and culture, respectively. We found no statistically significant differences in the reduction of viral shedding or time to clearance of virus between viral subtypes. Antiviral-resistant viruses were recovered from 3 (27.3%) of 11 children with influenza A subtype H1N1, 1 (2.9%) of 34 children with influenza A subtype H3N2, and 0 (0%) of 19 children with influenza B virus, all of whom were treated with oseltamivir (P = .004). There was no evidence of prolonged illness in children infected with drug-resistant virus. CONCLUSIONS: Drug resistance emerges at a higher rate in influenza A subtype H1N1 virus than in influenza A subtype H3N2 or influenza B virus after tiered weight-based oseltamivir therapy. Virological surveillance for patterns of drug resistance is essential for determination of antiviral treatment strategies and for composition of pandemic preparedness stockpiles. PMID- 19133797 TI - Low plasma level of cathelicidin antimicrobial peptide (hCAP18) predicts increased infectious disease mortality in patients undergoing hemodialysis. AB - BACKGROUND: Human cathelicidin antimicrobial protein (hCAP18) is an antimicrobial and immunomodulatory peptide that has pleiotropic effects and is transcriptionally regulated by vitamin D. Because the administration of vitamin D analogues has been linked to decreased mortality among patients with end-stage renal disease, we hypothesized that low hCAP18 levels would identify those who are at increased risk of death attributable to infection while undergoing hemodialysis. METHODS: We performed a case-control study nested in a prospective cohort of patients (n = 10,044) initiating incident hemodialysis. Case patients (n = 81) were those who died of an infectious disease within 1 year; control patients (n = 198) were those who survived at least 1 year while undergoing dialysis. RESULTS: Mean (+/-SD) baseline levels of hCAP18 in case patients and control patients were 539 +/- 278 ng/mL and 650 +/- 343 ng/mL, respectively (P = .006). hCAP18 levels had a modest correlation with 1,25-dihydroxyvitamin D levels r = 0.23; P = .053) but not with 25-hydroxyvitamin D levels r = -0.06; P = .44). Patients with hCAP18 levels in the lowest tertile had a 2-fold increased risk (odds ratio, 2.1; 95% confidence interval, 1.2-3.5) of death attributable to infection; after multivariable adjustment, this relationship remained statistically significant (odds ratio, 3.7; 95% confidence interval, 1.2-11.2). CONCLUSIONS: In individuals initiating chronic hemodialysis, low baseline levels of hCAP18, a vitamin D-regulated antimicrobial protein, are independently associated with an increased risk of death attributable to infection. PMID- 19133798 TI - Measurement of airborne influenza virus in a hospital emergency department. AB - Size-fractionated aerosol particles were collected in a hospital emergency department to test for airborne influenza virus. Using real-time polymerase chain reaction, we confirmed the presence of airborne influenza virus and found that 53% of detectable influenza virus particles were within the respirable aerosol fraction. Our results provide evidence that influenza virus may spread through the airborne route. PMID- 19133799 TI - Pregnancy and optimal care of HIV-infected patients. AB - Human immunodeficiency virus (HIV) infection during pregnancy is a condition that requires multidisciplinary care. Care must be rendered that is appropriate for both the mother and the fetus. Prevention of mother-to-child transmission of HIV is of paramount concern. To prevent transmission, universal testing for HIV infection in pregnant women is ideal. In the United States and other developed countries, great strides have been made toward decreasing the risk of HIV transmission to infants to <2% with use of a combination of highly active antiretroviral therapy during the antepartum period and during labor and delivery, scheduled cesarean section when appropriate, avoidance of breast feeding, and 6 weeks of zidovudine prophylaxis for infants. The continuation of antiretroviral therapy after delivery depends on the needs of the mother with regard to treatment of her own health. In resource-limited countries, where simplified and shortened courses of antiretroviral regimens have been used, reduction in mother-to-child transmission has also been shown, although not as effectively as that with highly active antiretroviral therapy. In these settings, exclusive breast-feeding for 6 months is recommended to reduce the risk of postnatal transmission. PMID- 19133800 TI - Single-dose nevirapine to prevent mother-to-child transmission of HIV type 1: balancing the benefits and risks. PMID- 19133801 TI - Nitazoxanide versus vancomycin in Clostridium difficile infection: a randomized, double-blind study. AB - BACKGROUND: Vancomycin is the only US Food and Drug Administration-approved drug for treatment of Clostridium difficile infection (CDI). Metronidazole has been widely used for this purpose but may be inferior to vancomycin, especially for hospitalized patients with severe disease. We report a prospective, double-blind, randomized controlled trial comparing nitazoxanide with vancomycin for treatment of CDI. METHODS: Fifty patients with CDI were randomized to receive vancomycin or nitazoxanide for 10 days. An initial response was considered to be the absence of all CDI symptoms between days 11 and 13, and a final response was considered to be lack of symptom recurrence by day 31. RESULTS: One patient fulfilled an exclusion criterion and was removed from the study. Twenty-seven patients received vancomycin, and 23 received nitazoxanide; 23 and 18 patients, respectively, completed the full course of treatment. Initial responses occurred in 20 (74%) of 27 patients treated with vancomycin and in 17 (77%) of 22 patients treated with nitazoxanide (95% confidence interval, -24% to +28%). In those who completed therapy, response rates were 87% (20 of 23 patients) in the vancomycin group and 94% (17 of 18 patients) in the nitazoxanide group (95% confidence interval, -18% to +30%). Times to complete resolution of symptoms were similar in the 2 groups (P = .55). Two patients in the vancomycin group and 1 patient in the nitazoxanide group experienced relapse within 31 days after beginning treatment. Sustained response rates were 78% (18 of 23 patients) for the vancomycin group, and 89% (16 of 18 patients) for the nitazoxanide group (95% confidence interval, 18% to +35%). CONCLUSIONS: The small sample precludes conclusions about noninferiority of nitazoxanide to vancomycin. Nevertheless, this is the first recent randomized controlled trial to compare any antimicrobial agent other than metronidazole with vancomycin. Results suggest that nitazoxanide may be as effective as vancomycin in treating CDI. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00384527. PMID- 19133802 TI - The role of breast-feeding in the prevention of Helicobacter pylori infection: a systematic review. AB - BACKGROUND: The benefits of breast-feeding for the prevention of infection in infants and young children have been widely recognized, but epidemiologic studies regarding the role of breast-feeding in protecting against Helicobacter pylori infection have produced conflicting results. METHODS: We performed a systematic review of relevant epidemiologic studies conducted during the period 1984-2007 after abstracting data from articles that met our inclusion criteria. Study quality was assessed using the Newcastle-Ottawa scale. With use of the random effects model, we calculated the summary odds ratios (ORs) and 95% confidence intervals (CIs) for H. pylori infection according to history of breast-feeding. RESULTS: For the 14 studies that met inclusion criteria, the summary OR for H. pylori infection was 0.78 (95% CI, 0.61-0.99; 1-sided P = .002). Nine of the 14 studies reported ORs of <1.0, and 6 of these studies reported statistically significant protective effects. Only 1 study reported a statistically significant OR of >1.0. In studies in which the subjects resided in middle- or low-income nations, the summary OR was 0.55 (95% CI, 0.33-0.93; P = .01), compared with 0.93 (95% CI, 0.73-1.19; P = .28) in studies in which subjects resided in high-income nations. The summary OR for studies that use the (13)C-urea breath test was 0.67 (95% CI, 0.32-1.39), compared with 0.91 (95% CI, 0.74-1.11) for studies that used the H. pylori IgG serologic test. We found no statistically significant dose dependent protective effect against H. pylori associated with increasing duration of breast-feeding. CONCLUSIONS: Breast-feeding is protective against H. pylori infection, especially in middle- and low-income nations. PMID- 19133803 TI - Severe postvaccinia encephalitis with acute disseminated encephalomyelitis: recovery with early intravenous immunoglobulin, high-dose steroids, and vaccinia immunoglobulin. AB - We report the second case of severe postvaccinial encephalitis with acute disseminated encephalomyelitis since smallpox vaccination was reintroduced in 2002. Both affected patients responded dramatically with early intervention of intravenous immunoglobulin. Our patient, who also received concurrent vaccinia immunoglobulin and corticosteroids, demonstrated full recovery. PMID- 19133804 TI - Persistent minority K103N mutations among women exposed to single-dose nevirapine and virologic response to nonnucleoside reverse-transcriptase inhibitor-based therapy. AB - OBJECTIVE: We investigated whether there are long-lasting effects of exposure to single-dose nevirapine (sdNVP) treatment on virologic response to nonnucleoside reverse-transcriptase inhibitor (NNRTI)-based therapy among human immunodeficiency virus (HIV)-infected women. METHODS: An observational epidemiologic study was conducted in Johannesburg, South Africa. Initial and sustained virologic response to NNRTI-based therapy was compared between 94 HIV infected women who had received sdNVP 18-36 months earlier and 60 unexposed, HIV infected women who had been pregnant 12-36 months earlier. Viral load was measured every 4 weeks up to week 24 and then every 12 weeks up to week 78. Time to viral suppression (viral load, <50 copies/mL) and confirmed rebound in the viral load (viral load, >400 copies/mL) were compared. Drug resistance was assessed using K103N allele-specific real-time polymerase chain reaction assay and population sequencing. RESULTS: Almost all women (97.5% of sdNVP-exposed women and 91.3% of sdNVP-unexposed women; P = .21) achieved viral suppression by week 24, and similar percentages of sdNVP-exposed and -unexposed women (19.4% and 15.1%, respectively) experienced viral rebound within 78 weeks after treatment (P = .57). K103N was detected with the K103N allele-specific real-time polymerase chain reaction assay among sdNVP-exposed and -unexposed women before treatment; detection was strongly predictive of inadequate viral response: 60.9% of women for whom K103N was detected in either viral RNA or DNA did not experience viral suppression or experienced viral rebound, compared with 15.1% of women for whom K103N was not detected (P < .001). After treatment, the M184V mutation occurred less frequently among sdNVP-exposed women than among sdNVP-unexposed women, but the frequency of NNRTI-associated mutations was similar between these groups of women with inadequate virologic response. CONCLUSIONS: Exposure to sdNVP in the prior 18-36 months was not associated with a reduced likelihood of achieving and sustaining viral suppression while receiving NNRTI-based therapy. However, women with minority K103N mutations before treatment had a reduced durability of virologic suppression. PMID- 19133806 TI - Immunization with a nontoxic mutant of staphylococcal enterotoxin A, SEAD227A, protects against enterotoxin-induced emesis in house musk shrews. AB - BACKGROUND: Staphylococcal enterotoxins (SEs) are the most common cause of foodborne diseases and toxic shock throughout the world. However, no vaccine that prevents emesis induced by SEs has been described. METHODS: A nontoxic mutant of SEA, SEAD227A, was constructed by site-directed mutagenesis and was purified by means of the Escherichia coli expression system. House musk shrews, a small emetic animal model, were immunized with SEAD227A and then challenged with wild type SEA. SEA-induced emesis was recorded for 3 h. Antibody production was analyzed by gel double-immunodiffusion assay. Neutralizing activities of the antibodies with respect to superantigenic and emetic activities were analyzed in vitro and in vivo. RESULTS: SEAD227A was devoid of both superantigenic and emetic activities but still retained its immunological activity. Immunization with SEAD227A strongly induced specific antibody production and provided significant protection against SEA-induced emesis. Antibodies from immunized shrews markedly inhibited the SEA-induced proliferation of spleen cells and also significantly ablated SEA-induced vomiting in the animals. CONCLUSIONS: These results suggest that vaccination with SEAD227A, which is devoid of toxic properties, provides protection against SEA-induced emesis. This nontoxic mutant and its specific antibodies might be useful in the prevention and treatment of staphylococcal food poisoning. PMID- 19133805 TI - Crystallographic, spectroscopic, and computational analysis of a flavin C4a oxygen adduct in choline oxidase. AB - Flavin C4a-OO(H) and C4a-OH adducts are critical intermediates proposed in many flavoenzyme reaction mechanisms, but they are rarely detected even by rapid transient kinetics methods. We observe a trapped flavin C4a-OH or C4a-OO(H) adduct by single-crystal spectroscopic methods and in the 1.86 A resolution X-ray crystal structure of choline oxidase. The microspectrophotometry results show that the adduct forms rapidly in situ at 100 K upon exposure to X-rays. Density functional theory calculations establish the electronic structures for the flavin C4a-OH and C4a-OO(H) adducts and estimate the stabilization energy of several active site hydrogen bonds deduced from the crystal structure. We propose that the enzyme-bound FAD is reduced in the X-ray beam. The aerobic crystals then form either a C4a-OH or C4a-OO(H) adduct, but an insufficient proton inventory prevents their decay at cryogenic temperatures. PMID- 19133807 TI - Mycobacterium avium paratuberculosis invades human small-intestinal goblet cells and elicits inflammation. AB - Crohn disease is a chronic inflammatory bowel disease of unknown etiology. Mycobacterium avium paratuberculosis (MAP) was found in the gut of patients with Crohn disease, but causality was not established. Fully developed, germ-free human small intestine and colon were established by subcutaneous transplantation of fetal gut into SCID (severe combined immunodeficiency) mice thereafter infected by direct intraluminal inoculation of MAP. We have found that MAP actively invades the human gut epithelial goblet cells of the small intestine, inducing severe tissue damage and inflammation. These observations indicate that MAP can specifically colonize the normal human small intestine and can elicit inflammation and severe mucosal damage. PMID- 19133808 TI - Factors associated with acquisition and clearance of human papillomavirus infection in a cohort of US men: a prospective study. AB - BACKGROUND: Our understanding of factors associated with acquisition and clearance of human papillomavirus (HPV) in men has been limited. This study sought to determine factors associated with those aspects of HPV infection in a cohort of US men. METHODS: A total of 285 men aged 18-44 years were monitored every 6 months for approximately 18 months. Risk-factor information was obtained at each visit by use of a self-administered questionnaire. A continuous-time 2 state Markov model was applied. RESULTS: Lifetime number of sex partners reported at enrollment was the most significant risk factor for acquisition of all types of HPV. Men reporting >16 lifetime sex partners were at significantly elevated risk of any HPV infection (adjusted hazard ratio [AHR], 2.8 [95% confidence interval {CI}, 1.1-7.1]), oncogenic HPV infection (AHR, 9.6 [95% CI, 2.4-37.8]), and nononcogenic HPV infection (AHR, 3.6 [95% CI, 1.3-9.9]), compared with those reporting 0-4 partners. Circumcised men were 3 and 6 times more likely to clear infection with any and oncogenic HPV types, respectively. In addition, having had >16 lifetime sex partners was associated with greater likelihood of clearance of oncogenic HPV infection (AHR, 4.9 [95% CI, 1.2-19.8]). CONCLUSION: The key factor associated with acquisition of HPV was lifetime number of sex partners, whereas circumcision was the most significant determinant for clearance of any HPV infection and oncogenic HPV infection. PMID- 19133809 TI - Worldwide epidemiology of neutralizing antibodies to adeno-associated viruses. AB - Recombinant adeno-associated viruses (AAVs) have unique gene-transfer properties that speak to their potential as carriers for gene therapy or vaccine applications. However, the presence of neutralizing antibodies to AAV as a result of previous exposure can significantly limit effective gene transfer. In this study, we obtained 888 human serum samples from healthy volunteers in 10 countries around the world. Samples were assayed for neutralizing antibodies to AAV1, AAV2, AAV7, and AAV8, as well as to a novel, structurally distinct AAV vector, rh32.33, in an in vitro transduction inhibition assay. Our data revealed that neutralizing antibodies to AAV2 were the most prevalent antibodies in all regions, followed by antibodies to AAV1. The seroprevalences of antibodies to AAV7 and to AAV8 were lower than that for antibodies to AAV1, and neutralization of AAVrh32.33 was only rarely detected. Our data also indicate a strong linkage of seroreactivity between apparently distinct serotypes that has not been predicted previously in animal models. PMID- 19133811 TI - Factors associated with fluctuations in IgA and IgG levels at the cervix during the menstrual cycle. AB - OBJECTIVE: The objective of this analysis was to describe patterns and determinants of cervical immunoglobulin A (IgA) and G (IgG) levels during the menstrual cycle. METHODS: A total of 154 women who attended 3 visits coinciding with the follicular, periovulatory, and luteal phases of their menstrual cycle were studied. Cervical secretions were collected at each visit for determination of total IgA and IgG levels. Questionnaires administered at each visit inquired about demographic characteristics and behavioral practices. RESULTS: Total IgA and IgG levels were higher among oral contraceptive (OC) users than among naturally cycling women (hereafter, "non-OC users"). IgA and IgG levels decreased at midcycle, particularly among non-OC users. After adjustment for phase of the current cycle, specimen weight, and detection of blood in the sample, report of a recent illness was associated with lower IgA and IgG levels and increased age with higher IgA and IgG levels among OC users and non-OC users. Increased lifetime number of pregnancies was associated with a higher IgA level among non OC users and a higher IgG level among OC users. Change in immunoglobulin levels between visits was associated with sample weight and the presence of blood for both OC users and non-OC users. CONCLUSIONS: Phase of the current menstrual cycle and OC use were significant determinants of cervical IgA and IgG levels. The impacts of endogenous and exogenous hormones on cervical immunoglobulin levels should be further investigated. PMID- 19133813 TI - Formation of crystalline polymers from the reaction of amine-functionalized C60 with silver salts. AB - Three new crystalline compounds of Ag(I) with fullerene-containing ligands, the piperidine adduct [C(60)(N(CH(2)CH(2))(2)N)] and the fullero[60]pyrrolidine [C(60)(CH(2)N(CH(3))CH(2))], have been prepared and characterized by X-ray crystallography. The polymeric structure of {[C(60)(N(CH(2)CH(2))(2)N)][Ag(O(2)CCF(3))](2)}.CS(2) consists of linear chains composed of two distinct molecules of the functionalized fullerene, with four Ag(I) ions attached to the four nitrogen atoms and four bridging trifluoroacetate ions. Two of the four Ag(I) ions form eta(2)-bonds to carbon atoms in different regions of the fullerene cage, and there is one close Ag-Ag contact (3.1657(7) A) as well. These chains are further cross-linked by bridging trifluoroacetate ions. The structure of {[C(60)(CH(2)N(CH(3))CH(2))]Ag(NO(3))}.0.25CH(3)OH involves two similar polymeric chains in which Ag(I) ions bind to the nitrogen atom of one N methyl-3,4-fullero[60]pyrrolidine ligand and to a carbon atom of another N-methyl 3,4-fullero[60]pyrrolidine in eta(1)-fashion for one chain and in distorted eta(2)-fashion in the other. Additionally, each Ag(I) ion is bonded to two oxygen atoms from two bridging nitrate ions. On the other hand, [C(60)(N(CH(2)CH(2))(2)N)](2)Ag(NO(3)).0.5CH(3)OH.CH(2)Cl(2) is a simple coordination complex with two very large ligands attached to Ag(I). Coordination of Ag(I) to C(60) produces much smaller alterations to the fullerene geometry than does coordination of Pt(0)(PPh(3))(2) or Ir(I)(CO)Cl(PPh(3))(2) groups. PMID- 19133814 TI - A simple model to describe the effect of electrostatic interactions on the composition of mixed self-assembled monolayers. AB - Due to intermolecular interactions, the surface composition of mixed self assembled monolayers often differs markedly from the solution composition. In the case of charged surfactants, large deviations from the ideality have been reported. The effect of the electrostatic interactions between charged compounds on the surface composition is examined using a simple gas lattice model, in the Bragg-Williams approximation. The interaction potential is obtained from the Debye-Huckel treatment of electrolytic solutions. The model is able to explain and reproduce the deviations observed experimentally, both qualitatively and quantitatively. The key role played by the ionic strength on the surface composition is emphasized. PMID- 19133810 TI - Phylogenetic reconstruction of transmission events from individuals with acute HIV infection: toward more-rigorous epidemiological definitions. AB - Phylogenetic reconstructions of transmission events from individuals with acute human immunodeficiency virus (HIV) infection are conducted to illustrate this group's heightened infectivity. Varied definitions of acute infection and assumptions about observed phylogenetic clusters may produce misleading results. We conducted a phylogenetic analysis of HIV pol sequences from 165 European patients with estimated infection dates and calculated the difference between dates within clusters. Nine phylogenetic clusters were observed. Comparison of dates within clusters revealed that only 2 could have been generated during acute infection. Previous analyses may have incorrectly assigned transmission events to the acutely HIV infected when they were more likely to have occurred during chronic infection. PMID- 19133819 TI - Acquisition and cross-transmission of Staphylococcus aureus in European intensive care units. AB - OBJECTIVE: To study the acquisition and cross-transmission of Staphylococcus aureus in different intensive care units (ICUs). METHODS: We performed a multicenter cohort study. Six ICUs in 6 countries participated. During a 3-month period at each ICU, all patients had nasal and perineal swab specimens obtained at ICU admission and during their stay. All S. aureus isolates that were collected were genotyped by spa typing and multilocus variable-number tandem repeat analysis typing for cross-transmission analysis. A total of 629 patients were admitted to ICUs, and 224 of these patients were found to be colonized with S. aureus at least once during ICU stay (22% were found to be colonized with methicillin-resistant S. aureus [MRSA]). A total of 316 patients who had test results negative for S. aureus at ICU admission and had at least 1 follow-up swab sample obtained for culture were eligible for acquisition analysis. RESULTS: A total of 45 patients acquired S. aureus during ICU stay (31 acquired methicillin susceptible S. aureus [MSSA], and 14 acquired MRSA). Several factors that were believed to affect the rate of acquisition of S. aureus were analyzed in univariate and multivariate analyses, including the amount of hand disinfectant used, colonization pressure, number of beds per nurse, antibiotic use, length of stay, and ICU setting (private room versus open ICU treatment). Greater colonization pressure and a greater number of beds per nurse correlated with a higher rate of acquisition for both MSSA and MRSA. The type of ICU setting was related to MRSA acquisition only, and the amount of hand disinfectant used was related to MSSA acquisition only. In 18 (40%) of the cases of S. aureus acquisition, cross-transmission from another patient was possible. CONCLUSIONS: Colonization pressure, the number of beds per nurse, and the treatment of all patients in private rooms correlated with the number of S. aureus acquisitions on an ICU. The amount of hand disinfectant used was correlated with the number of cases of MSSA acquisition but not with the number of cases of MRSA acquisition. The number of cases of patient-to-patient cross-transmission was comparable for MSSA and MRSA. PMID- 19133820 TI - Research implications of pigment biology in zebrafish. PMID- 19133821 TI - Skin color in fish and humans: impacts on science and society. AB - As genetic model systems, fish have played a key role in our understanding of a wide range of biological processes, including vertebrate pigmentation. In this review, we focus on one aspect of pigmentation, skin pigmentation, which has been of momentous importance in human history. Two fish models, medaka and zebrafish, played important roles in demystifying skin color and, by extension, the concept of "race." Related thinking has the potential to make two additional contributions to human welfare. Fish can be used to validate gene candidates from genome-wide association studies (GWAS) in what has been called "Systems Genetics." Because fish are familiar vertebrates, and share genetic mechanisms of skin color with humans, they also have outstanding potential as an educational tool-to "demystify" race, to increase public understanding of the role of model systems and evolution in science, and to enhance appreciation of both genetic and environmental factors that impact human health and society. PMID- 19133822 TI - U.S. scientists' role in the eugenics movement (1907-1939): a contemporary biologist's perspective. PMID- 19133824 TI - Neocuproine ablates melanocytes in adult zebrafish. AB - The simplest regeneration experiments involve the ablation of a single cell type. While methods exist to ablate the melanocytes of the larval zebrafish,(1,2) no convenient method exists to ablate melanocytes in adult zebrafish. Here, we show that the copper chelator neocuproine (NCP) causes fragmentation and disappearance of melanin in adult zebrafish melanocytes. Adult melanocytes expressing eGFP under the control of a melanocyte-specific promoter also lose eGFP fluorescence in the presence of NCP. We conclude that NCP causes melanocyte death. This death is independent of p53 and melanin, but can be suppressed by the addition of exogenous copper. NCP is ineffective at ablating larval melanocytes. This now provides a tool for addressing questions about stem cells and the maintenance of the adult pigment pattern in zebrafish. PMID- 19133823 TI - Melanoma biology and the promise of zebrafish. AB - Advantageous organismal and technical attributes of the zebrafish are being increasingly applied to study cancer biology. Along with other tumor models, zebrafish that develop melanomas have been generated. In both genetics and phenotype, zebrafish melanomas are strikingly similar to their human counterparts. For this reason, studies in the zebrafish are poised to make significant contributions to melanoma biology. In this review, we summarize important features of human melanoma and discuss how the zebrafish can be used to address many questions that remain unanswered about this devastating disease. PMID- 19133825 TI - Natural variation of male ornamental traits of the guppy, Poecilia reticulata. AB - Male ornamental traits of the guppy, Poecilia reticulata, provide an outstanding example of natural variation in sex-linked male-advantageous traits that are shaped by both sexual and environmental selection. A substantial fraction of the underlying genes is known to be genetically linked to the sex-determining region on the differentiating Y-chromosome. Intercrosses between parental populations originating from geographically distant locations in East Trinidad and Cumana (Venezuela) were used to study segregation of ornamental traits in male progeny. In addition, we performed backcrosses to compare segregation of ornaments in presence or absence of prominent traits linked to the Y-chromosome. Another backcross strategy involving XY females from the laboratory strain zebrinus maculatus allowed studying additive and dominant effects of alleles on two different Y-chromosomes on pattern formation. For genetic mapping, we have previously developed nuclear SNP markers linked to expressed genes, including several genes known to be important for pattern formation in other species. Of these candidate genes 15 were placed on 11 different linkage groups. Our phenotypic and genotypic analysis of progeny from mapping crosses and backcrosses suggests several genetic mechanisms that enhance natural variation, namely, additive effects of codominant alleles, suppressive actions of dominant alleles, and a complex interplay between sex-linked and autosomal cofactors. PMID- 19133827 TI - mc1r Pathway regulation of zebrafish melanosome dispersion. AB - Zebrafish rapidly alter their pigmentation in response to environmental changes. For black melanocytes, this change is due to aggregation or dispersion of melanin in the cell. Dispersion and aggregation are controlled by intracellular cyclic adenosine monophosphate (cAMP) levels, which increase upon stimulation by alpha melanocyte-stimulating hormone (alpha-MSH) or reduce with melanin-concentrating hormone (MCH). In mammals and birds, the melanocortin-1-receptor (MC1R) responds to MSH, and stimulates the synthesis of black eumelanin. While MSH-cAMP signaling stimulates melanogenesis in mammals, and melanosome dispersal in cold-blood vertebrates, the pathway components are highly conserved. However, it has only been assumed that mc1r mediates melanosome dispersal in fish. Here, using morpholino oligonucleotides designed to knockdown mc1r expression, we find that mc1r morphants are unable to disperse melanosomes when grown in dark conditions. We also use chemical modifiers of the cAMP pathway, and find an unexpected response to the specific phosphodiesterase 4 (PDE4) inhibitor, rolipram, in melanosome dispersal. When treated with the drug, melanosomes fail to fully disperse in dark conditions, despite presumed increased levels of cAMP, and in contrast to the effects of the nonselective PDE inhibitor, 3-isobutyl-1 methylxanthine. In conclusion, we demonstrate a direct role for mc1r in zebrafish melanosome dispersal in response to background, and use chemical modification of this pathway to uncover a possible new layer of regulation in melanosome dispersal in zebrafish. PMID- 19133826 TI - Small molecule modifier screen for kit-dependent functions in zebrafish embryonic melanocytes. AB - Zebrafish is gaining popularity as a vertebrate model for screening small molecules that affect specific phenotypes or genetic pathways. In this study, we present a targeted drug screen to identify drug modifiers of the melanocyte migration defect of a temperature-sensitive allele of the Kit receptor tyrosine kinase, kit(ts). We first test two candidate drugs, the phosphatidylinositol-3 kinase kinase inhibitor (LY294002) and the Erk/MAP kinase inhibitor (PD98059), for their effect on melanocyte migration and survival. We find that LY294002 enhances the migration defect of kit(ts), implicating the phosphatidylinositol-3 kinase kinase pathway in promoting kit-dependent melanocyte migration, but not survival. We then used the kit(ts)-sensitized genetic background to screen a panel of 1280 pharmacologically active drugs to identify drug enhancers and suppressors of the kit(ts) melanocyte migration defect. We identified three drug enhancers of migration, two of which, Papaverine and Isoliquiritigenin, specifically enhance the kit(ts) migration defect, while 8-DPAT affected both melanocyte migration and survival. These drugs now provide additional experimental tools for investigating the mechanisms of kit-promoted melanocyte migration and survival in the zebrafish embryo. PMID- 19133828 TI - Pigments, patterns, and fish behavior. AB - Color patterns in fish are often multicomponent signals, composed of pigment based and structural color patches that can be used to communicate within species, in both inter- and intrasexual interactions, and between species. In this review, we discuss some of the roles played by pigment-based elements of color pattern. We begin by discussing general forms of coloration, classifying them by appearance (e.g., cryptic vs. conspicuous) and apparent function (e.g., conspicuous coloration and mating displays, stripes and cooperation, and bars and aggression). We then briefly discuss the roles pigments play in the perception of these color patterns via their presence in the eye. In the last section, we look at the relative importance of carotenoid versus melanic coloration in situations where honest signals to potential rivals and potential mates might be required. In this survey, we have highlighted some recent research, especially studies that consider both the physiological and behavioral processes underlying the evolution and expression of pigment-based color patterns in fish. The nature of pigmented color patterns depends not just on the dynamics of pattern development and physiological regulation, but also on the behavioral roles played by these patterns, both now and in the past. As such, advances in particular fields of study on pigment patterns (physiology, developmental biology, behavioral ecology, evolutionary biology, etc.) will increasingly depend on insights from other fields. PMID- 19133829 TI - Pigment dilution mutants from fish models with connection to lysosome-related organelles and vesicular traffic genes. AB - An interesting question in developmental biology is why mutations in genes with functions essential for the majority of cells produce diseases affecting only specific tissues. For example, pigment dilution disorders are often the consequence of mutations in conserved vesicular traffic genes. In Hermansky Pudlak, Griscelli, and Chediak-Higashi pigment dilution syndromes, vesicular traffic mutations affect several organs with one characteristic in common: to carry out their functions they depend to a great extent on lysosome-related organelles (LROs), such as the melanosomes in melanocytes. Conserved multimeric complexes, present in most cell types, target proteins to lysosomes or selected LROs using transport vesicles. By studying these diseases or the model organisms that are defective in these processes, we have learned that every cell type possesses a unique way to regulate its vesicular traffic machinery and to assemble its multimeric complexes. This is accomplished by subunits from these multimeric complexes acting in a cell-specific manner. Here, we review several fish pigment dilution mutants that represent models for human vesicular traffic diseases. PMID- 19133830 TI - Functional evaluation of isolated zebrafish hearts. AB - Traditional working heart preparations, based on the original Langendorff setup, are widely used experimental models that have tremendously advanced the cardiovascular field. However, these systems can be deceivingly complex, requiring the maintenance of pH with CO(2), the delivery of oxygenated perfusate, and the need for extensive laboratory equipment. We have examined the feasibility of using isolated zebrafish (Danio rerio) hearts as an experimental model system, in which experimental procedures can be performed in the absence of the traditional requirements and sophisticated setup equipment. Isolated zebrafish hearts exhibited spontaneous contractile activity, could be electrically paced, and were responsive to pharmacologic stimulation with isoproterenol for 1.5 h after in vivo removal. Isolated zebrafish hearts offer a time- and cost-effective alternative to traditional Langendorff/working heart preparation models, and could be used to investigate cardiac function and repair. PMID- 19133831 TI - Zebrafish as a new model for herpes simplex virus type 1 infection. AB - The zebrafish (Danio rerio) is rapidly gaining ground as a disease model. However, until now, the use of this species with human pathogens has been restricted to just three bacteria; no studies involving viruses that infect humans are recorded. In this study, the zebrafish was used as a model of herpes simplex virus type 1 (HSV-1) infection of the nervous system. Fish infected using viral culture supernatants showed detectable HSV-1 DNA concentrations 1-4 days after inoculation, indicating that this virus can experimentally infect and persist in this host. The kinetics of infection was dose dependent, especially in the head. Histological immunodetection of HSV-1 glycoproteins confirmed the presence of HSV-1 in the organs studied; infection led to histopathological changes. Moreover, the suppression of the immune system by cyclophosphamide and the antiviral effect of acyclovir were demonstrated. The infection of the encephalon was studied in detail, and the time course of viral colonization recorded. Immunofluorescence studies provided immunoreactive evidence of viral antigens in the encephalon and spinal cord. Viruses cleared from infected brains showed the ability to infect human neuroblastoma cells. This study is the first to demonstrate HSV-1 infection in the zebrafish and manifests the potential use of this species in herpesvirus studies. PMID- 19133832 TI - The effect of a depth gradient on the mating behavior, oviposition site preference, and embryo production in the zebrafish, Danio rerio. AB - Captive zebrafish (Danio rerio) exhibit a limited repertoire of mating behaviors, likely due to the somewhat unnatural environment of aquaria. Observations in their natural habitat led us to believe that a depth gradient within the mating setup would positively affect fish mating. By tilting the tank to produce a depth gradient, we observed novel behaviors along with a preference for oviposition in the shallow area. Although we did not see an increase in the likelihood of a pair of fish to mate, we did see an increase in the embryo output in both adults and juveniles. In the adults, tilting led to a significant increase in embryo production (436 +/- 35 tilted vs. 362 +/- 34 untilted; p < 0.05). A similar effect was seen in juvenile fish as they progressed through sexual maturity. These results suggest that tilting of mating cages in the laboratory setting will lead to demonstrable improvements in embryo production for zebrafish researchers, and highlights the possibility of other manipulations to increase fecundity. PMID- 19133835 TI - Thermoresponsive block copolymers of poly(ethylene glycol) and polyphosphoester: thermo-induced self-assembly, biocompatibility, and hydrolytic degradation. AB - Novel thermoresponsive block copolymers of poly(ethylene glycol) and polyphosphoester were synthesized, and the thermo-induced self-assembly, biocompatibility, and hydrolytic degradation behavior were studied. The block copolymers with various molecular weights and compositions were synthesized through ring-opening polymerization of 2-ethoxy-2-oxo-1,3,2-dioxaphospholane (EEP) and 2-isopropoxy-2-oxo-1,3,2-dioxaphospholane (PEP) using poly(ethylene glycol) monomethyl ether (mPEG) as the initiator and stannous octoate as the catalyst. The obtained block polymers exhibited thermo-induced self-assembly behavior, demonstrated by dynamic light scattering and UV-vis measurements using 1,6-diphenyl-1,3,5-hexatriene as the probe. It was found that the critical aggregation temperature (CAT) of the block copolymers shifted to higher temperature with increased molecular weight of mPEG, while copolymerization with more hydrophobic monomer PEP led to lower transition temperature; thus, the CAT can be conveniently adjusted. The block copolymers did not induce significant hemolysis and plasma protein precipitation. In vitro MTT and live/dead staining assays indicated they are biocompatible, and the biocompatibility was further demonstrated in vivo by the absence of local acute inflammatory response in mouse muscle following intramuscular injection. Unlike most frequently studied thermoresponsive poly(N-isopropylacrylamide), polyphosphoesters were hydrolytically degradable in aqueous solution that was proven by gel permeation chromatography and NMR analyses, and the degradation products were proven to be nontoxic to HEK293 cells. Therefore, with good biocompatibility and thermoresponsiveness, these biodegradable block copolymers of mPEG and polyphosphoesters are promising as stimuli-responsive materials for biomedical applications. PMID- 19133836 TI - Synthesis and water absorption transport mechanism of a pH-sensitive polymer network structured on vinyl-functionalized pectin. AB - Polysaccharide-structured copolymer hydrogel having excellent pH-sensitivity was developed from N,N-dimethylacrylamide (DMAc) and vinyl-functionalized Pectin (Pec). The Pec was vinyl-functionalized by way of chemical reaction with glycidyl metacrylate (GMA) in water under acidic and thermal stimuli. 13C NMR, 1H NMR, and FT-IR spectra revealed that the vinyl groups coming from the GMA were attached onto backbone of the polysaccharide. The hydrogels were obtained by polymerization of the Pec-vinyl with the DMAc. 13C-CP/MAS NMR and FTIR spectra confirmed that the gelling process occurred by way of the vinyl groups attached on Pec-vinyl backbone. The values of apparent swelling rate constant (k) decreased appreciably for pH greater than 6, demonstrating the swelling process of the hydrogel becomes slower at more alkaline conditions. There was an increase of diffusional exponent (n) with increasing pH of the surrounding liquid. This means the water absorption profile becomes more dependent on the polymer relaxation in basified swelling media. In this condition, a longer water absorption half-time (t1/2) was verified, suggesting the polymer relaxation mechanism of the hydrogel would have a considerable effect on the t1/2. PMID- 19133837 TI - This issue introduces a set of ideas called "Distributed Drug Discovery (D(3))". PMID- 19133838 TI - One-pot synthesis of quinazolinone derivatives from nitro-compounds with the aid of low-valent titanium. AB - The synthesis of a series of quinazolinone derivatives such as 2 thioxoquinazolinones, imidazo[1,2-c]quinazolin-5-amines, and benzimidazo[1,2 c]quinazolin-5-amines, starting from nitro-compounds has been described. This general approach features a easy way for access to the target quinazolinone derivatives. The key cyclization step embraces the formation of a thiourea intermediate, mediated by low-valent titanium, and the other important intermediate was also obtained. A variety of substrates can participate in the process with good yields, making this methodology suitable for library synthesis in drug discovery efforts. PMID- 19133839 TI - Catalyst screening for oxidative reforming of methane in direct route using high pressure HTS reactor with syngas detection system by colorimetric reaction and gas chromatograph. AB - A high-throughput screening (HTS) reactor for high-pressure oxidative reforming of methane in a direct reaction route was developed. With a combination of catalyst preparation by a split-and-pool method and HTS, Ni-K/alpha-Al(2)O(3) catalyst was found to show high activity under 1 MPa at 650 degrees C with high selectivity even when O(2) conversion is less than 100%. The HTS reactor required a new simple syngas detector operable under high pressure because the number of parallel reactor is limited when equipped with the conventional detection system. The complexity of the pressure reducing unit is the main reason of the limitation. Reduction of metal oxide accompanied with the color change was applied to the detection system. Copper oxide was supported on the filter disk made of alumina, and the filter was placed underneath the catalyst bed. After the methane was oxidatively reformed under 1 MPa at 650 degrees C, color change of spots from dark brown to light brown was observed just under the catalyst which produced hydrogen. Color change of the disk can be used to detect hydrogen formation from the reforming catalyst under pressure. PMID- 19133840 TI - The origin, evolution and structure of the protein world. AB - Contemporary protein architectures can be regarded as molecular fossils, historical imprints that mark important milestones in the history of life. Whereas sequences change at a considerable pace, higher-order structures are constrained by the energetic landscape of protein folding, the exploration of sequence and structure space, and complex interactions mediated by the proteostasis and proteolytic machineries of the cell. The survey of architectures in the living world that was fuelled by recent structural genomic initiatives has been summarized in protein classification schemes, and the overall structure of fold space explored with novel bioinformatic approaches. However, metrics of general structural comparison have not yet unified architectural complexity using the 'shared and derived' tenet of evolutionary analysis. In contrast, a shift of focus from molecules to proteomes and a census of protein structure in fully sequenced genomes were able to uncover global evolutionary patterns in the structure of proteins. Timelines of discovery of architectures and functions unfolded episodes of specialization, reductive evolutionary tendencies of architectural repertoires in proteomes and the rise of modularity in the protein world. They revealed a biologically complex ancestral proteome and the early origin of the archaeal lineage. Studies also identified an origin of the protein world in enzymes of nucleotide metabolism harbouring the P-loop-containing triphosphate hydrolase fold and the explosive discovery of metabolic functions that recapitulated well-defined prebiotic shells and involved the recruitment of structures and functions. These observations have important implications for origins of modern biochemistry and diversification of life. PMID- 19133842 TI - Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. AB - Calreticulin is an ER (endoplasmic reticulum) luminal Ca2+-buffering chaperone. The protein is involved in regulation of intracellular Ca2+ homoeostasis and ER Ca2+ capacity. The protein impacts on store-operated Ca2+ influx and influences Ca2+-dependent transcriptional pathways during embryonic development. Calreticulin is also involved in the folding of newly synthesized proteins and glycoproteins and, together with calnexin (an integral ER membrane chaperone similar to calreticulin) and ERp57 [ER protein of 57 kDa; a PDI (protein disulfide-isomerase)-like ER-resident protein], constitutes the 'calreticulin/calnexin cycle' that is responsible for folding and quality control of newly synthesized glycoproteins. In recent years, calreticulin has been implicated to play a role in many biological systems, including functions inside and outside the ER, indicating that the protein is a multi-process molecule. Regulation of Ca2+ homoeostasis and ER Ca2+ buffering by calreticulin might be the key to explain its multi-process property. PMID- 19133843 TI - A case for education in palliative and end-of-life care in emergency medicine. PMID- 19133841 TI - Repair of ionizing radiation-induced DNA double-strand breaks by non-homologous end-joining. AB - DNA DSBs (double-strand breaks) are considered the most cytotoxic type of DNA lesion. They can be introduced by external sources such as IR (ionizing radiation), by chemotherapeutic drugs such as topoisomerase poisons and by normal biological processes such as V(D)J recombination. If left unrepaired, DSBs can cause cell death. If misrepaired, DSBs may lead to chromosomal translocations and genomic instability. One of the major pathways for the repair of IR-induced DSBs in mammalian cells is NHEJ (non-homologous end-joining). The main proteins required for NHEJ in mammalian cells are the Ku heterodimer (Ku70/80 heterodimer), DNA-PKcs [the catalytic subunit of DNA-PK (DNA-dependent protein kinase)], Artemis, XRCC4 (X-ray-complementing Chinese hamster gene 4), DNA ligase IV and XLF (XRCC4-like factor; also called Cernunnos). Additional proteins, including DNA polymerases mu and lambda, PNK (polynucleotide kinase) and WRN (Werner's Syndrome helicase), may also play a role. In the present review, we will discuss our current understanding of the mechanism of NHEJ in mammalian cells and discuss the roles of DNA-PKcs and DNA-PK-mediated phosphorylation in NHEJ. PMID- 19133844 TI - Prevalence of occult bacteremia in children aged 3 to 36 months presenting to the emergency department with fever in the postpneumococcal conjugate vaccine era. AB - OBJECTIVES: The goal of this study was to identify the prevalence of occult bacteremia (OB) in well-appearing, previously healthy children aged 3 to 36 months who present to the emergency department (ED) with fever without source in the post-pneumococcal conjugate vaccine (PCV) era. METHODS: This was a retrospective cohort study of children presenting to an urban pediatric ED between July 1, 2004, and June 30, 2007. Children were included if they were aged 3 to 36 months, febrile, and previously healthy; had no source of infection on examination; had a blood culture drawn; and were discharged from the ED. Outcome measures were rates of OB and contaminant rates. RESULTS: A total of 8,408 children met all inclusion criteria. There were 21 true-positives, yielding an OB rate of 0.25% (95% confidence interval [CI] = 0.16% to 0.37%). There were 159 contaminant cultures yielding a contaminant rate of 1.89% (95% CI = 1.61% to 2.19%), or a ratio of 7.6 contaminants for each true-positive. There were 14 included patients who grew Streptococcus pneumoniae from the blood, for a rate of 0.17% (95% CI = 0.09% to 0.27%). CONCLUSIONS: Given the current rate of OB in the post-PCV era, it may no longer be cost-effective to send blood cultures on well appearing, previously healthy children aged 3 to 36 months who have fever without source. PMID- 19133846 TI - Emergent aspirin use in cardiovascular disease in the emergency department: oral dosing versus rectal suppositories. PMID- 19133847 TI - Tuesdays to write ... A guide to time management in academic emergency medicine. PMID- 19133848 TI - Post-traumatic stress disorder symptomology associated with witnessing unsuccessful out-of-hospital cardiopulmonary resuscitation. AB - OBJECTIVES: The objective was to assess symptoms of post-traumatic stress disorder (PTSD) associated with witnessing unsuccessful out-of-hospital cardiopulmonary resuscitation (CPR) on a family member. METHODS: Adult family members of deceased, adult, nontraumatic out-of-hospital cardiac arrest victims who were transported to a large, Midwestern hospital were contacted by telephone beginning 1 month after the event. Subjects were dichotomized as to whether or not they were physically present during the patient's resuscitation. A structured interview obtained the patient's prearrest functioning, whether the family member witnessed or performed CPR, patient and family demographic data, key cardiac arrest events, and a measure of subject PTSD symptoms (PTSD Symptom Scale Interview [PSS-I]). RESULTS: There were 34 witnesses and 20 nonwitnesses. Each group was similar in race, religion, age, gender, and relationship to the patient. Patients in each group were similar in prearrest functioning. Witnesses' total PTSD symptom scores were nearly two times higher than nonwitnesses (14.47 vs. 7.60, respectively; mean difference = 6.87, 95% confidence interval [CI] = 0.57 to 13.17). Two PSS-I subscales were higher for witnesses than nonwitnesses: Avoidance (5.41 vs. 2.25; mean difference = 3.16, 95% CI = 0.74 to 5.58) and Increased Arousal (4.26 vs. 2.20; mean difference = 2.06, 95% CI = 0.08 to 4.05), while Reexperiencing was not (4.79 vs. 3.15; mean difference = 1.64, 95% CI = 0.62 to 3.91). Linear regression analysis indicated that witnessing CPR of a loved one was associated with a mean increase of nearly 12 points on the PSS-I after controlling for the possibility of other potentially influential events and characteristics. Results were similar when CPR providers (n = 6) were removed from the witness group. CONCLUSIONS: Witnessing a failed CPR attempt of a loved one in an out-of-hospital location may be associated with displaying symptoms of PTSD in the early term of the bereavement period. While preliminary, these data suggest that the relationship exists even after controlling for other potential factors that may also affect the propensity for displaying such symptoms, such as the suddenness and location of the patient's cardiac arrest. PMID- 19133849 TI - A picture's worth. PMID- 19133850 TI - North Carolina Disease Event Tracking and Epidemiologic Collection Tool (NC DETECT) and the National Hospital Ambulatory Medical Care Survey (NHAMCS): comparison of emergency department data. AB - The North Carolina Disease Event Tracking and Epidemiologic Collection Tool (NC DETECT) is a near-real-time database of emergency department (ED) visits automatically extracted from hospital information system(s) in the state of North Carolina. The National Hospital Ambulatory Medical Care Survey (NHAMCS) is a retrospective probability sample survey of visits to U.S. hospital EDs. This report compares data from NC DETECT (2006) with NHAMCS (2005) ED visit data to determine if the two data sets are consistent. Proportions, rates, and confidence intervals (CIs) were calculated for ED visits by age and gender; arrival method and age; expected source of payment; disposition; hospital admissions; NHAMCS top 20 diagnosis groups and top five primary diagnoses by age group; International Classifications of Disease, 9th revision, Clinical Modification (ICD-9-CM) primary diagnosis codes; and cause of injury. North Carolina DETECT captured 79% of statewide ED visits. Twenty-eight persons for every 100 North Carolina residents visited a North Carolina ED that reports to NC DETECT at least once in 2006, compared to 20% nationally. Twenty-seven percent of ED visits in North Carolina had private insurance as the expected payment source, compared with 40% nationwide. The proportion of injury-related ED visits in North Carolina is 25%, compared to 36.4% nationally. Rates and proportions of disease groups are similar. Similarity of NC DETECT rates and proportions to NHAMCS provides support for the face and content validity of NC DETECT. The development of statewide near real-time ED databases is an important step toward the collection, aggregation, and analysis of timely, population-based data by state, to better define the burden of illness and injury for vulnerable populations. PMID- 19133852 TI - Diagnosis of pulmonary embolism using emergency department bedside echocardiogram. PMID- 19133854 TI - The Centers for Medicare and Medicaid Services (CMS) community-acquired pneumonia core measures lead to unnecessary antibiotic administration by emergency physicians. AB - OBJECTIVES: The objectives were to assess emergency physician (EP) understanding of the Centers for Medicare and Medicaid Services (CMS) core measures for community-acquired pneumonia (CAP) guidelines and to determine their self reported effect on antibiotic prescribing patterns. METHODS: A convenience sample of EPs from five medical centers in North Carolina was anonymously surveyed via a Web-based instrument. Participants indicated their level of understanding of the CMS CAP guidelines and the effects on their prescribing patterns for antibiotics. RESULTS: A total of 121 EPs completed the study instrument (81%). All respondents were aware of the CMS CAP guidelines. Of these, 95% (95% confidence interval [CI] = 92% to 98%) correctly understood the time-based guidelines for antibiotic administration, although 24% (95% CI = 17% to 31%) incorrectly identified the onset of this time period. Nearly all physicians (96%; 95% CI = 93% to 99%) reported institutional commitment to meet these core measures, and 84% (95% CI = 78% to 90%) stated that they had a department-based CAP protocol. More than half of the respondents (55%; 95% CI = 47% to 70%) reported prescribing antibiotics to patients they did not believe had pneumonia in an effort to comply with the CMS guidelines, and 42% (95% CI = 34% to 50%) of these stated that they did so more than three times per month. Only 40% (95% CI = 32% to 48%) of respondents indicated a belief that the guidelines improve patient care. Of those, this was believed to occur by increasing pneumonia awareness (60%; 95% CI = 52% to 68%) and improving hospital processes when pneumonia is suspected (86%; 95% CI = 80% to 92%). CONCLUSIONS: Emergency physicians demonstrate awareness of the current CMS CAP guidelines. Most physicians surveyed reported the presence of institutional protocols to increase compliance. More than half of EPs reported that they feel the guidelines led to unnecessary antibiotic usage for patients who are not suspected to have pneumonia. Only 40% of EPs believe that CAP awareness and expedient care resulting from these guidelines has improved overall pneumonia-related patient care. Outcome-based data for non-intensive care unit CAP patients are lacking, and EPs report that they prescribe antibiotics when they may not be necessary to comply with existing guidelines. PMID- 19133855 TI - Correlation of plasma and peritoneal diasylate clomipramine concentration with hemodynamic recovery after intralipid infusion in rabbits. AB - OBJECTIVES: Drug sequestration to an expanded plasma lipid phase has been proposed as a potential mechanism of action for lipid emulsions in lipophilic cardiotoxin overdose. The authors set out to document plasma and peritoneal diasylate clomipramine concentration after resuscitation with lipid emulsion in a rabbit model of clomipramine-induced hypotension. METHODS: Twenty sedated mechanically ventilated New Zealand White rabbits were allocated to receive either 12 mL/kg 20% Intralipid or 12 mL/kg saline solution, following clomipramine infusion to 50% baseline mean arterial pressure (MAP). Hemodynamic parameters and serum clomipramine concentration were determined to 59 minutes. Peritoneal dialysis with 20% Intralipid or saline solution was evaluated for clomipramine concentration. RESULTS: Mean arterial pressure was greater in lipid treated animals as assessed by repeated-measures analysis of variance (F[1,14] = 6.84; p = 0.020). Lipid infusion was associated with elevated plasma clomipramine concentration and reduced initial volume of distribution (Vd; 5.7 [+/-1.6] L/kg lipid vs. 15.9 [+/-7.2] L/kg saline; p = 0.0001). Peritoneal diasylate clomipramine concentration was greater in lipid-treated animals (366.2 [+/-186.2] microg/L lipid vs. 37.7 [+/-13.8] microg/L saline; p = 0.002). CONCLUSIONS: Amelioration of clomipramine-induced hypotension with lipid infusion is associated with reduced initial Vd and elevated plasma clomipramine concentration consistent with intravascular drug-lipid sequestration. Concomitant peritoneal dialysis with lipid emulsion enhances clomipramine extraction. PMID- 19133856 TI - There may be no room in the inn, but the innkeepers sure are happy. PMID- 19133857 TI - Letters on "Predictors of mortality in patients with delerium tremens". PMID- 19133859 TI - Letters on "Predictors of mortality in patients with delerium tremens". PMID- 19133860 TI - My two cents on radiation. PMID- 19133862 TI - Functional changes of the cortical motor system in hereditary spastic paraparesis. AB - BACKGROUND: Hereditary spastic paraparesis (HSP) is a heterogeneous group of disorders characterized by progressive bilateral lower limb spasticity. Functional imaging studies in patients with corticospinal tract involvement have shown reorganization of motor circuitry. Our study investigates functional changes in sensorimotor brain areas in patients with HSP. METHODS: Twelve subjects with HSP and 12 healthy subjects were studied. Functional magnetic resonance imaging (fMRI) was used to measure brain activation during right-hand finger tapping. Image analysis was performed using general linear model and regions of interest (ROI)-based approach. Weighted laterality indices (wLI) and anterior/posterior indicies (wAI and wPI) were calculated for predefined ROIs. RESULTS AND DISCUSSION: Comparing patients and controls at the same finger tapping rate (1.8 Hz), there was increased fMRI activation in patients' bilateral posterior parietal cortex and left primary sensorimotor cortex. No differences were found when comparing patients and controls at 80% of their individual maximum tapping rates. wLI of the primary sensorimotor cortex was significantly lower in patients. Subjects with HSP also showed a relative increase in the activation of the posterior parietal and premotor areas compared with that of the primary sensorimotor cortex. Our findings demonstrate an altered pattern of cortical activation in subjects with HSP during motor task. The increased activation probably reflects reorganization of the cortical motor system. PMID- 19133863 TI - Congenital myopathies--a comprehensive update of recent advancements. AB - The congenital myopathies are relatively newly discovered compared with other categories of muscle diseases. Current research continues to clarify and classify the congenital myopathies. These pose a diagnostic problem and cannot be diagnosed by routine hematoxylin and eosin stain. A lot of special techniques are required to diagnose them correctly and it's various subtypes. The disease specific structural changes seen in the muscle are detected by enzyme histochemistry, immunohistochemistry and electron microscopy. Through this review we provide an up-to-date analysis of congenital myopathies including clinical and pathologic aspects. PMID- 19133864 TI - Post-polio syndrome: epidemiologic and prognostic aspects in Brazil. AB - OBJECTIVES: To describe the clinical and epidemiological aspects of post-polio syndrome (PPS) and identify predictors of its severity. MATERIALS AND METHODS: 132 patients with PPS were selected at the Neuromuscular Disease Outpatient Clinic of the Federal University of Sao Paulo. Descriptive analysis was carried out and predictors of PPS severe forms were investigated using an unconditional logistic regression. RESULTS: The average age at onset was 39.4 years. The most common symptoms were fatigue (87.1%), muscle pain (82.4%) and joint pain (72.0%); 50.4% of the cases were severe. The following were associated with PPS severity: a < or =4-year period of neurological recovery (OR 2.8), permanent damage in two limbs (OR 3.6) and residence at the time of acute polio in a city with more advanced medical assistance (OR 2.5). CONCLUSIONS: Health professionals should carefully evaluate polio survivors for PPS and be aware of the implications of muscle overuse in the neurological recovery period. PMID- 19133866 TI - Feasibility and logistics of MRI before thrombolytic treatment. AB - OBJECTIVES: The study analyzes feasibility and time-delays in Magnetic resonance imaging (MRI) based thrombolysis and estimate the impact of MRI on individual tissue plasminogen activator (rtPA) treatment. MATERIALS AND METHODS: Feasibility of MRI and time logistics were prospectively recorded in patients referred with presumed acute stroke over a 2 year time period. Door-to-needle-times (DNT) were compared with those of patients treated with rtPA after conventional CT during the same time period, and to published open label studies. RESULTS: We received 174 patients with presumed stroke. MRI was feasible in 141 of 161 (88%) of those requiring acute imaging. MRI supported the decision to treat 11 patients with mild symptoms or seizures, and not to treat four patients with extensive infarctions. Median 'door-to-needle time' (DNT) in MR scanned patients (70 min), did not differ significantly from DNT after conventional CT (n = 17, DNT = 66 min, P = 0.27) or the Safe Implementation of Thrombolysis in Stroke (SITS-MOST) registry (DNT = 68 min). CONCLUSIONS: Magnetic resonance imaging can be performed in the majority of acute stroke patients without delaying treatment. MRI may affect decision making in a large proportion of patients. PMID- 19133865 TI - Conversion from valproic acid onto topiramate in adolescents and adults with epilepsy. AB - OBJECTIVE: To explore efficacy and tolerability outcomes of topiramate (TPM) in patients with epilepsy transitioning from valproic acid (VPA) because of insufficient efficacy and/or tolerability onto TPM. METHODS: Multicenter, open label, single arm, non-interventional study examining patients (> or =12 years) with epilepsy, transitioning onto TPM from baseline mono-or combination therapy with VPA. TPM was added onto the existing antiepileptic drug (AED) treatment and started at a dose of 25 mg once daily. The dose was titrated up with 25 mg/day increments, once every 1-2 weeks, until a final dose between 50-200 mg/day was reached. Based on clinical judgment, the treating physician decided whether or not and when the existing AED treatment especially with VPA could be withdrawn. Documented were type and frequency of seizures, TPM dose, quality of life (QOLIE 10 questionnaire), subjective perception of improvement, and adverse events (AE). RESULTS: One hundred and forty-seven patients (59% women, mean age 41 years) switched from baseline VPA treatment onto TPM because of insufficient efficacy (61%) and/or poor tolerability (81%). Average duration of follow-up was 20.3 weeks with an overall discontinuation rate of 16.3% of patients, mainly because of AE (in 8.2% of 147 patients). At study endpoint, the intended shift to TPM monotherapy was achieved in 70% of patients at a median dose of 150 mg/day. A seizure reduction of > or =50% was achieved in 75% of patients in the last scheduled period (week 8-20), and 51% of patients entering that period remained seizure-free. Quality of life improved significantly as compared with baseline for all domains of QOLIE-10 (P < 0.001). Most frequent AEs were weight decrease (4.8%), paraesthesia and fatigue (4.1% each), speech disorder and headaches (2.7% each). CONCLUSIONS: In patients with epilepsy not satisfactorily treated with VPA, conversion to TPM was associated with improved seizure control as well as improvement in several aspects of quality of life. PMID- 19133867 TI - Breast-feeding and a subsequent diagnosis of measles. AB - BACKGROUND: Breast-feeding protects against many infectious diseases and may also influence immunization outcomes. AIM: This study investigated if breast-feeding protects against clinical measles and if it modified the effect of immunization. METHODS: We used logistic regression with data for 10 207 individuals from the 1970 British Cohort study (BCS70). Breast-feeding data were collected at five years of age, and information on clinical measles infection, as well as socio economic measures was collected at the age of ten years. Breast feeding was categorized as: breast-fed <1 month (n = 1611), breast-fed for 1-3 months (n = 1016), breast-fed for more than three months (n = 1108), breast-feeding of uncertain duration (n = 21) and never breast-fed (n = 6451). RESULTS: Breast feeding for more than three months was negatively associated with a diagnosis of clinical measles infection after adjustment for crowding, social class, measles vaccination, parity and sex with an odds ratio (95% confidence interval) of 0.69 (0.60-0.81) compared with those who never breast-fed. Measles vaccination was highly associated with low risk for measles with: 0.14 (0.13-0.16). Age at acute measles infection was not associated with breastfeeding. Breast-feeding did not notably alter measles immunization efficacy. CONCLUSION: Immunization against measles provides effective protection against the disease. A more modest reduction in the risk of a measles diagnosis is associated with breast-feeding. The associations with a diagnosis of measles for breast-feeding and measles immunization are independent of each other. PMID- 19133868 TI - Group A streptococcal throat infection - to treat or not to treat? PMID- 19133869 TI - Endemic measles in Karachi, Pakistan and validation of IMCI criteria for measles. AB - AIM: To estimate the incidence of measles in Karachi, Pakistan and to determine the proportion of children with measles based on the WHO integrated management of childhood illness (IMCI) criteria with a positive IgM for measles or dengue. METHODS: Patients up to 14 years old were screened for febrile rash illnesses at five Karachi hospitals. Active measles cases were classified as measles, measles with eye and mouth complications, or severe complicated measles using IMCI criteria. RESULTS: Screening 1,219,061 patients over a 39-month period identified 3503 qualified children. Most (76.8%) measles cases occurred in children under five years of age. The average annual incidence rate was 0.68 per 1000 in year 1; 0.19 in year 2 and 0.08 in year 3 of surveillance. Pneumonia and diarrhoea were the most common complications. Of 18.1% hospitalized, 1.6% died. Of 2286 children tested, 1599 (69.9%) were measles IgM positive. Of 542 measles IgM negative children, 66 (12%) were dengue IgM positive. The predictive positive value for the IMCI case definition was 75%. CONCLUSION: The IMCI case definitions for measles is reasonable but may overestimate measles incidence. Measles continues to be a public health problem in Pakistan; increased efforts to control measles are urgently needed. PMID- 19133870 TI - Trends in outcomes for very preterm infants in the southern region of Sweden over a 10-year period. AB - AIM: To investigate trends in mortality and morbidity in very preterm infants. METHODS: Population-based perinatal register; liveborn infants 22 + 0 to 31 + 6 gestational weeks were investigated (time period 1995-2004). Time trends for mortality and common morbidities were explored using logistic regression analyses. RESULTS: Data from 1614 liveborn infants were included. There was an increase in live born infants below 25 gestational weeks, annual odds ratio (OR) 1.15 (95% CI: 1.08-1.23) and a decrease in mortality annual OR 0.82 (95% CI: 0.69 0.98). The rates of bronchopulmonary dysplasia (BPD) and sepsis increased during the study period, annual ORs of 1.10 (95% CI: 1.04-1.17) and 1.09 (95% CI: 1.03 1.16). The duration of mechanical ventilation increased for surviving infants <25 gestational weeks (p = 0.003), while the duration of continuous positive airway pressure (CPAP) increased for infants <28 gestational weeks (p = <0.001). There were no changes in the rates of intraventricular haemorrhages (IVH, 3-4), retinopathy of prematurity (ROP, 3-5), seizures or necrotizing enterocolitis (NEC). CONCLUSION: During the 10-year period changes in mortality and morbidity were most pronounced for infants with GA <28 gestational weeks. The increasing rate of sepsis was present in infants <28 gestational weeks, whereas the increase in BPD was demonstrated in the whole study population <32 gestational weeks. PMID- 19133871 TI - The 1998 Nobel Prize--discovery of the role of nitric oxide as a signalling molecule. PMID- 19133872 TI - Cardiorespiratory fitness and body mass index values in 9-year-old rural Norwegian children. AB - AIM: To describe cardiorespiratory fitness and body mass index (BMI) values in a representative population of 9-year-old Norwegian children in two rural communities and compare present values with previous findings. METHODS: Two hundred and fifty-nine 9-year-old children were invited, and 256 participated in this study. Maximal oxygen uptake was directly measured during a continuous progressive treadmill protocol. Body mass and height were also measured. RESULTS: The mean +/- SD relative maximal oxygen uptake was 52.8 +/- 6.5 for boys and 46.9 +/- 7.2 mL/kg/min for girls. Eight percent of the boys and 16.8% of the girls were classified as overweight, and 1.6% of the boys and 6.9% of the girls as obese. Mean age, body mass, height and Ponderal index were not significantly different between sexes. Girls had a higher BMI than boys (p = 0.05). CONCLUSION: Compared to earlier Norwegian studies, children's BMI values seem to have increased substantially. This increase is most pronounced in girls. When assessing these differences using the PI, this increase is less marked. Comparing maximal oxygen uptake data with that in earlier Nordic studies, there is no evidence that fitness has declined among 9-year olds. However, the limitations of the few earlier studies make reliable comparisons difficult. PMID- 19133873 TI - Effects of zinc supplementation on parent and teacher behaviour rating scores in low socioeconomic level Turkish primary school children. AB - OBJECTIVE: To determine the effect of zinc supplementation on behaviour in low income school aged children. DESIGN: Double-blind randomized, placebo controlled trial. SETTING: Low-income district primary school in Turkey. PARTICIPANTS: Third grade students in the school. Among 252 students, 226 participated and 218 completed the study. INTERVENTION: Children in each class were randomized either to the study group to receive 15 mg/day elemental zinc syrup or to placebo group to receive the syrup without zinc for 10 weeks. MAIN OUTCOME MEASURES: The change in Conner's Rating Scales for Teachers and Parents scores after supplementation. RESULTS: The mean Conner's Rating Scale for Parents scores on attention deficit, hyperactivity, oppositional behaviour and conduct disorder decreased significantly in the study and placebo groups after supplementation (p < 0.01). The prevalence of children with clinically significant parent ratings on attention deficit (p = 0.01) and hyperactivity (p = 0.004) decreased in the study group while prevalence of oppositional behaviour (p = 0.007) decreased in the placebo group. In children of mothers with low education all mean Parents' scores decreased significantly (p < 0.01) in the study group while only hyperactivity scores decreased in the placebo group (p < 0.01). In this subgroup the prevalence of children with clinically significant scores for attention deficit, hyperactivity and oppositional behaviour decreased only in the study group (p < 0.05). There was no change in mean Teachers' scores. CONCLUSION: In our study zinc supplementation decreased the prevalence of children with clinically significant scores for attention deficit and hyperactivity. The affect on behaviour was more evident in the children of low educated mothers. PMID- 19133874 TI - B(2) kinin receptors mediate the Indian red scorpion venom-induced augmentation of visceral reflexes via the nitric oxide cyclic guanosine monophosphate pathway. AB - AIM: This study was performed to delineate the kinin (receptor)-dependent pathways in the Indian red scorpion (Mesobuthus tamulus; MBT) venom-induced pulmonary oedema as well as the augmentation of cardio-pulmonary reflexes evoked by phenyldiguanide (PDG). METHODS: In urethane-anaesthetized adult rats, the effect of venom on the PDG reflex responses (blood pressure, heart rate and respiration rate) and the pulmonary water content was ascertained using various antagonists(des- Arg, B(1) receptor antagonist; Hoe 140, B(2) receptor antagonist; N(omega)-nitro-l-arginine methyl ester (l-NAME), nitric oxide (NO) synthase inhibitor; methylene blue, soluble guanylate cyclase inhibitor; and glibenclamide, K(+)(ATP) channel blocker). The effect of phosphodiesterase V inhibitor (sildenafil citrate) on the reflex response and the pulmonary water content was also examined and compared with venom-induced responses. RESULTS: Intravenous injection of PDG (10 microg kg(-1)) evoked apnoea, bradycardia and hypotension lasting >60 s. Exposure to MBT venom (100 microg kg(-1)) for 30 min augmented the PDG reflex responses by two times and increased the pulmonary water content, significantly. Hoe 140 blocked the venom-induced responses (augmentation of PDG reflex and increased pulmonary water content) whereas des-Arg did not. l NAME, methylene blue or glibenclamide also blocked the venom-induced responses. Furthermore, sildenafil citrate (that increases cGMP levels) produced augmentation of PDG reflex response and increased the pulmonary water content as seen with venom. CONCLUSION: The results indicate that venom-induced responses involve B(2) kinin receptors via the NO-dependent guanylate cyclase-cGMP pathway involving K(+)(ATP) channels. PMID- 19133875 TI - Association of depressive symptoms and metabolic syndrome in men. AB - OBJECTIVE: To explore the relationship between several indicators of depression and metabolic syndrome (MetS). METHOD: A population-based sample with high (HMS group) or low (LMS group) levels of mental symptoms, including those of depression, in three follow-ups participated in a clinical examination in 2005 (n = 223). MetS was determined according to the NCEP criteria. RESULTS: The prevalence of MetS was 49% in men and 21% in women. Men with MetS had higher rates of major depressive disorder than other men. They also displayed higher Hamilton Rating Scale for Depression (HDRS) scores and more often signs of suicidality. In logistic regression analyses, higher HDRS scores (OR 1.31, 95% CI 1.04-1.64) and belonging to the HMS group (OR 10.1, 95% CI 1.98-51.3) were independent associates for MetS but only in men. CONCLUSION: The results highlight that there is an association between long-term depressive symptoms and the emergence of MetS, especially in men. PMID- 19133876 TI - Cognition as predictor of current and follow-up depressive symptoms in the general population. AB - OBJECTIVE: Previous studies have reported an association between depression and poor cognitive functioning. Unknown is to what degree such associations are merely state-related or reflect an enduring depression vulnerability. This study examined whether cognitive deficits predict current and/or follow-up (sub)clinical depressive symptoms in the general population. METHOD: A population based sample of 569 female twins and 43 of their sisters completed a neuropsychological battery. Cross-sectional and prospective associations between depressive symptoms measured at the subclinical [Symptom Checklist-90 (SCL-90)] and clinical level (Structured Clinical Interview for DSM-IV disorders) and neuropsychological factors (episodic memory and information processing speed) were examined. RESULTS: Structured Clinical Interview for DSM-IV disorders baseline depressive symptoms were significantly associated with information processing speed but not with episodic memory. Episodic memory was significantly associated with follow-up SCL-90 depressive symptoms. CONCLUSION: Being depressed is accompanied by slower information processing. Poor memory functioning may be a predictor for the onset of subclinical depressive symptoms. PMID- 19133877 TI - A cross-sectional investigation of discontinuation of self-injury and normalizing pain perception in patients with borderline personality disorder. AB - OBJECTIVE: Several studies have shown reduced pain perception in patients with borderline personality disorder (BPD) and current self-injurious behavior (SIB). The aim of the present study was to test whether pain perception in patients with current SIB is different from that of patients who had stopped SIB, and whether pain perception of the latter group differs from healthy controls (HC). METHOD: We investigated 24 borderline patients and 24 HC. Thirteen patients showed current SIB (BPD-SIB) and 11 patients did not exhibit SIB anymore (BPD-non-SIB). Pain thresholds were assessed using thermal stimuli and laser radiant heat pulses. RESULTS: We found significant linear trends for all pain measures. The BPD-SIB group was less sensitive than the BPD-non-SIB group and the latter were less sensitive than HC. The pain sensitivity negatively correlated with borderline symptom severity. CONCLUSION: The results suggest an association between the termination of SIB, decline of psychopathology and normalization of pain perception in borderline patients. PMID- 19133878 TI - A pilot examination of social context and everyday physical activity among adults receiving Community Mental Health Services. AB - OBJECTIVE: Community mental health center (CMHC) clients include a variety of people with moderate to severe mental illnesses who also report a number of physical health problems. Physical activity (PA) has been identified as one intervention to improve health among this population; however, little is known about the role of social context in PA. The purpose of this study was to examine the role of social context in everyday PA among CMHC clients. METHOD: Data were collected from CMHC clients in two cultures using accelerometery and experience sampling methods. Data were analyzed using hierarchical linear modeling. RESULTS: Independence in housing nor culture was significantly associated with levels of PA. Being alone was significantly negatively related to PA level. CONCLUSION: Social isolation appears to be negatively related to PA at the level of everyday life. Physical activity interventions with this population should consider including social components as a part of PA. PMID- 19133879 TI - New dimensions in human laboratory models of addiction. PMID- 19133881 TI - Outlet density: a new frontier for tobacco control. PMID- 19133882 TI - Policy implications of the widespread practice of 'pre-drinking' or 'pre-gaming' before going to public drinking establishments: are current prevention strategies backfiring? AB - AIM: To describe the research, policy and prevention implications of pre-drinking or pre-gaming; that is, planned heavy drinking prior to going to a public drinking establishment. METHODS: The authors describe the phenomenon of pre drinking, motivations for pre-drinking and its associated risks using available research literature, media and popular internet vehicles. RESULTS: Heavy drinking prior to going out has emerged as a common and celebrated practice among young adults around the world. Apparent motivations are: (i) to avoid paying for high priced drinks at commercial drinking establishments; (ii) to achieve drunkenness and enhance and extend the night out; and (iii) to socialize with friends, reduce social anxiety or enhance male group bonding before going out. Limited existing research on pre-drinking suggests that it is associated with heavy drinking and harmful consequences. We argue that policies focused upon reducing drinking in licensed premises may have the unintended consequence of displacing drinking to pre-drinking environments, possibly resulting in greater harms. CONCLUSIONS: Effective policy and prevention for drinking in licensed premises requires a comprehensive approach that takes into account the entire drinking occasion (not just drinking that occurs in the licensed environment), as well as the 'determined drunkenness' goal of some young people. PMID- 19133883 TI - Does it matter where the drinking is, when the object is getting drunk? PMID- 19133885 TI - Coming off tranquillizers: a Sisyphean toil. PMID- 19133886 TI - Age-period-cohort modelling of alcohol volume and heavy drinking days in the US National Alcohol Surveys: divergence in younger and older adult trends. AB - AIMS: The decomposition of trends in alcohol volume and heavy drinking days into age, period, cohort and demographic effects offers an important perspective on the dynamics of change in alcohol use patterns in the United States. DESIGN: The present study utilizes data from six National Alcohol Surveys conducted over the 26-year period between 1979 and 2005. Setting United States. MEASUREMENTS: Alcohol volume and the number of days when five or more and eight or more drinks were consumed were derived from overall and beverage-specific graduated frequency questions. RESULTS: Trend analyses show that while mean values of drinking measures have continued to decline for those aged 26 and older, there has been a substantial increase in both alcohol volume and 5+ days among those aged 18-25 years. Age-period-cohort models indicate a potential positive cohort effect among those born after 1975. However, an alternative interpretation of an age-cohort interaction where drinking falls off more steeply in the late 20s than was the case in the oldest surveys cannot be ruled out. For women only, the 1956-60 birth cohort appears to drink more heavily than those born just before or after. Models also indicate the importance of income, ethnicity, education and marital status in determining these alcohol measures. CONCLUSIONS: Increased heavy drinking among young adults in recent surveys presents a significant challenge for alcohol policy and may indicate a sustained increase in future US alcohol consumption. PMID- 19133888 TI - The relative efficacy of two levels of a primary care intervention for family members affected by the addiction problem of a close relative: a randomized trial. AB - OBJECTIVES: A randomized trial to compare two levels of an intervention (full versus brief) for use by primary health-care professionals with family members affected by the problematic drug or alcohol use of a close relative. DESIGN: A prospective cluster randomized comparative trial of the two interventions. SETTING: A total of 136 primary care practices in two study areas within the West Midlands and the South West regions of England. PARTICIPANTS: A total of 143 family members affected by the alcohol or drug problem of a relative were recruited into the study by primary health-care professionals. All recruited family members were seen on at least one occasion by the professional delivering the intervention and 129 (90 %) were followed-up at 12 weeks. MAIN OUTCOME MEASURES: Two validated and standardized self-completion questionnaires measuring physical and psychological symptoms of stress (Symptom Rating Test) and behavioural coping (Coping Questionnaire) experienced by the family members. It was predicted that the full intervention would show increased reduction in both symptoms and coping when compared to the brief intervention. RESULTS: The primary analysis adjusted for clustering, baseline symptoms and stratifying variables (location and professional group) showed that there were no significant differences between the two trial arms. The symptom score at follow-up was 0.23 [95% confidence interval (CI): -3.65, +4.06] higher in the full intervention arm than in the brief intervention arm, and the coping score at follow-up was 0.12 (95% CI: -5.12, +5.36) higher in the full intervention arm than in the brief intervention arm. CONCLUSIONS: A well-constructed self-help manual delivered by a primary care professional may be as effective for family members as several face to-face sessions with the professional. PMID- 19133887 TI - Effects of alcoholism severity and smoking on executive neurocognitive function. AB - AIMS: Neurocognitive deficits in chronic alcoholic men are well documented. Impairments include memory, visual-spatial processing, problem solving and executive function. The cause of impairment could include direct effects of alcohol toxicity, pre-existing cognitive deficits that predispose towards substance abuse, comorbid psychiatric disorders and abuse of substances other than alcohol. Cigarette smoking occurs at higher rates in alcoholism and has been linked to poor cognitive performance, yet the effects of smoking on cognitive function in alcoholism are often ignored. We examined whether chronic alcoholism and chronic smoking have effects on executive function. METHODS: Alcoholism and smoking were examined in a community-recruited sample of alcoholic and non alcoholic men (n = 240) using standard neuropsychological and reaction-time measures of executive function. Alcoholism was measured as the average level of alcoholism diagnoses across the study duration (12 years). Smoking was measured in pack-years. RESULTS: Both alcoholism and smoking were correlated negatively with a composite executive function score. For component measures, alcoholism was correlated negatively with a broad range of measures, whereas smoking was correlated negatively with measures that emphasize response speed. In regression analyses, both smoking and alcoholism were significant predictors of executive function composite. However, when IQ is included in the regression analyses, alcoholism severity is no longer significant. CONCLUSIONS: Both smoking and alcoholism were related to executive function. However, the effect of alcoholism was not independent of IQ, suggesting a generalized effect, perhaps affecting a wide range of cognitive abilities of which executive function is a component. On the other hand, the effect of smoking on measures relying on response speed were independent of IQ, suggesting a more specific processing speed deficit associated with chronic smoking. PMID- 19133889 TI - Early pubertal maturation and drug use: underlying mechanisms. PMID- 19133890 TI - Incarceration and drug use patterns among a cohort of injection drug users. AB - AIMS: Drug law enforcement remains the dominant response to drug-related harm. However, the impact of incarceration on deterring drug use remains under evaluated. We sought to explore the relationship between incarceration and patterns of drug use among people who inject drugs (IDU). DESIGN: Using generalized estimating equations (GEE), we examined the prevalence and correlates of injection cessation among participants in the Vancouver Injection Drug User Study followed over 9 years. In subanalyses, we used McNemar's tests and linear growth curve analyses to assess changes in drug use patterns before and after a period of incarceration among participants reporting incarceration and those not incarcerated. FINDINGS: Among 1603 IDU, 842 (53%) reported injection cessation for at least 6 months at some point during follow-up. In multivariate GEE analyses, recent incarceration was associated negatively with injection cessation [adjusted odds ratio (AOR) = 0.43, 95% confidence interval (CI) 0.37-0.50], whereas the use of methadone was associated positively with cessation (AOR = 1.38, 95% CI 1.22-1.56). In subanalyses assessing longitudinal patterns of drug use among incarcerated individuals and those not incarcerated over the study period, linear growth curve analyses indicated that there were no statistically significant differences in patterns of drug use between the two groups (all P > 0.05). CONCLUSIONS: These observational data suggest that incarceration does not reduce drug use among IDU. Incarceration may inhibit access to mechanisms that promote injection cessation among IDU. In contrast, results indicate that methadone use is associated positively with injection cessation, independent of previous frequency of drug use. PMID- 19133891 TI - Ketamine use, cognition and psychological wellbeing: a comparison of frequent, infrequent and ex-users with polydrug and non-using controls. AB - INTRODUCTION: Preliminary research has indicated that recreational ketamine use may be associated with marked cognitive impairments and elevated psychopathological symptoms, although no study to date has determined how these are affected by differing frequencies of use or whether they are reversible on cessation of use. In this study we aimed to determine how variations in ketamine use and abstention from prior use affect neurocognitive function and psychological wellbeing. METHOD: We assessed a total of 150 individuals: 30 frequent ketamine users, 30 infrequent ketamine users, 30 ex-ketamine users, 30 polydrug users and 30 controls who did not use illicit drugs. Cognitive tasks included spatial working memory, pattern recognition memory, the Stockings of Cambridge (a variant of the Tower of London task), simple vigilance and verbal and category fluency. Standardized questionnaires were used to assess psychological wellbeing. Hair analysis was used to verify group membership. RESULTS: Frequent ketamine users were impaired on spatial working memory, pattern recognition memory, Stockings of Cambridge and category fluency but exhibited preserved verbal fluency and prose recall. There were no differences in the performance of the infrequent ketamine users or ex-users compared to the other groups. Frequent users showed increased delusional, dissociative and schizotypal symptoms which were also evident to a lesser extent in infrequent and ex-users. Delusional symptoms correlated positively with the amount of ketamine used currently by the frequent users. CONCLUSIONS: Frequent ketamine use is associated with impairments in working memory, episodic memory and aspects of executive function as well as reduced psychological wellbeing. 'Recreational' ketamine use does not appear to be associated with distinct cognitive impairments although increased levels of delusional and dissociative symptoms were observed. As no performance decrements were observed in the ex-ketamine users, it is possible that the cognitive impairments observed in the frequent ketamine group are reversible upon cessation of ketamine use, although delusional symptoms persist. PMID- 19133892 TI - Substance use disorder among older adults in the United States in 2020. AB - AIMS: This study aimed to project the number of people aged 50 years or older with substance use disorder (alcohol/illicit drug dependence or abuse) in the United States in 2020. DESIGN: Logistic regression models were applied to estimate parameters predicting past-year substance use disorder using the 2002-06 National Survey on Drug Use and Health data. We applied these parameters to the projected US 2020 population to estimate the number of adults aged 50 or older with substance use disorder in 2020. SETTING: Non-institutionalized US residences. PARTICIPANTS: Representative sample of the US civilian, non institutionalized population. MEASUREMENTS: Substance use disorder is classified based on criteria in the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. FINDINGS: Due to the large population size and high substance use rate of the baby-boom cohort, the number of adults aged 50 or older with substance use disorder is projected to double from 2.8 million (annual average) in 2002-06 to 5.7 million in 2020. Increases are projected for all examined gender, race/ethnicity and age groups. CONCLUSIONS: Our estimates provide critical information for policymakers to allocate resources and develop prevention and treatment approaches to address future needs of the US older adult population with substance use disorder. PMID- 19133894 TI - Hospitalizations for opioid poisoning: a nation-wide population-based study in Denmark, 1998-2004. AB - AIMS: To assess hospitalization rates (HR) for poisoning with heroin, methadone or strong analgesics and relate them to quantities of prescribed methadone and strong analgesics in Denmark between 1998 and 2004. DESIGN: Population-based ecological study. SETTINGS: We extracted data on all emergency department visits and hospital admissions registered in the Danish National Patient Registry with a diagnosis of poisoning with heroin (n = 1688), methadone (n = 173) or strong analgesics (n = 384). To ascertain sale of prescribed medications we used data from the Danish Medicines Agency. MEASUREMENTS: Age- and gender-standardized HR and defined daily doses (DDD) per 1000 people per day. FINDINGS: HR for heroin poisoning was 4.4 [95% confidence interval (CI): 3.8-4.9] per 100,000 person years (p-y) in 1998 and 4.6 (CI: 4.0-5.2) per 100,000 p-y in 2004. HR for methadone poisoning increased from 0.1 (CI: 0.0-0.2) per 100,000 p-y in 1998 to 1.1 (CI: 0.8-1.4) per 100,000 p-y in 2004. HR for poisoning with strong analgesics increased from 0.6 (CI: 0.4-0.9) per 100,000 p-y in 1998 to 2.1 (CI: 1.8-2.6) per 100,000 p-y in 2004. The sale of prescribed strong analgesics (5.0 DDD per 1000 people per day in 1998 to 5.9 DDD in 2004) and methadone (3.0 DDD per 1000 people per day in 1998 to 3.4 DDD in 2004) increased slightly between 1998 and 2004. CONCLUSION: Increasing sale of prescribed methadone and strong analgesics coincided with increasing HRs of poisoning with these drugs, whereas HR of heroin poisoning varied. Further longitudinal studies are important for the guidance of future policy making. PMID- 19133893 TI - New onsets of substance use disorders in borderline personality disorder over 7 years of follow-ups: findings from the Collaborative Longitudinal Personality Disorders Study. AB - AIMS: The purpose of this study was to examine whether patients with borderline personality disorder (BPD) have a higher rate of new onsets of substance use disorders (SUD) than do patients with other personality disorders (OPD). DESIGN: This study uses data from the Collaborative Longitudinal Personality Disorder Study (CLPS), a prospective naturalistic study with reliable repeated measures over 7 years of follow-up. SETTING: Multiple clinical sites in four northeastern US cities. PARTICIPANTS: A total of 175 patients with BPD and 396 patients with OPD (mean age 32.5 years) were assessed at baseline and at 6, 12, 24, 36, 48, 60, 72 and 84 months. MEASUREMENTS: The Structured Clinical Interview for DSM-IV Axis I Disorders and the Diagnostic Interview for DSM-IV Personality Disorders were used at baseline, the Follow-Along version of the DIPD-IV and the Longitudinal Interval Follow-up Evaluation at the follow-up evaluations. Kaplan-Meier analyses were calculated to generate the time to new onsets. FINDINGS: BPD patients showed a shorter time to new onsets of SUD. Thirteen per cent of BPD patients developed a new alcohol use disorder and 11% developed a new drug use disorder, compared to rates of 6% and 4%, respectively, for OPD. Non-remitted BPD and remitted BPD patients did not differ significantly in rates of new onsets of SUD. CONCLUSIONS: BPD patients have a high vulnerability for new onsets of SUDs even when their psychopathology improves. These findings indicate some shared etiological factors between BPD and SUD and underscore the clinical significance of treating SUD when it co-occurs in BPD patients. PMID- 19133895 TI - Randomized controlled trial of a brief intervention for problematic prescription drug use in non-treatment-seeking patients. AB - AIMS: Dependence on or problematic use of prescription drugs (PD) is estimated to be between 1 and 2% in the general population. In contrast, the proportion of substance-specific treatment in PD use disorders at 0.5% is comparatively low. With an estimated prevalence of 4.7%, PD-specific disorders are widespread in general hospitals compared to the general population. Brief intervention delivered in general hospitals might be useful to promote discontinuation or reduction of problematic prescription drug use. DESIGN: A randomized, controlled clinical trial. SETTING: Internal, surgical and gynaecological wards of a general and a university hospital. PARTICIPANTS: One hundred and twenty-six patients fulfilling criteria for either regular use of PD (more than 60 days within the last 3 months) or dependence on or abuse of PD, respectively, were allocated randomly to two conditions. INTERVENTION: Subjects received two counselling sessions based on Motivational Interviewing plus an individualized written feedback (intervention group, IG) or a booklet on health behaviour (control group, CG). MEASUREMENTS: The outcome was measured as reduction (>25%) and discontinuation of PD intake in terms of defined daily dosages (DDD). FINDINGS: After 3 months, more participants in the IG reduced their DDD compared to the participants in the CG (51.8% versus 30%; chi(2) = 6.17; P = 0.017). In the IG 17.9%, in the CG 8.6% discontinued use of PD (chi(2) = 2.42; P = 0.17). Conclusions Brief intervention based on Motivational Interviewing is effective in reducing PD intake in non-treatment-seeking patients. PMID- 19133897 TI - The clinical utility of genetic tests. PMID- 19133896 TI - Multiple genetic tests for susceptibility to smoking do not outperform simple family history. AB - AIMS: To evaluate the utility of using predictive genetic screening of the population for susceptibility to smoking. METHODS: The results of meta-analyses of genetic association studies of smoking behaviour were used to create simulated data sets using Monte Carlo methods. The ability of the genetic tests to screen for smoking was assessed using receiver operator characteristic curve analysis. The result was compared to prediction using simple family history information. To identify the circumstances in which predictive genetic testing would potentially justify screening we simulated tests using larger numbers of alleles (10, 15 and 20) that varied in prevalence from 10 to 50% and in strength of association [relative risks (RRs) of 1.2-2.1]. RESULTS: A test based on the RRs and prevalence of five susceptibility alleles derived from meta-analyses of genetic association studies of smoking performed similarly to chance and no better than the prediction based on simple family history. Increasing the number of alleles from five to 20 improved the predictive ability of genetic screening only modestly when using genes with the effect sizes reported to date. CONCLUSIONS: This panel of genetic tests would be unsuitable for population screening. This situation is unlikely to be improved upon by screening based on more genetic tests. Given the similarity with associations found for other polygenic conditions, our results also suggest that using multiple genes to screen the general population for genetic susceptibility to polygenic disorders will be of limited utility. PMID- 19133898 TI - Smoking, nicotine dependence and mental health among young adults: a 13-year population-based longitudinal study. AB - AIMS: To investigate prospectively the associations between daily smoking and nicotine dependence and anxiety, depression and suicide attempts. METHODS: Data were from the Young in Norway Longitudinal Study. A population-based sample (n = 1501) was followed for 13 years from ages 13-27 years. Data were gathered on smoking patterns and nicotine dependence; and depression, anxiety and parasuicide. Extensive information on socio-demographic factors, parental and family conditions, parental rearing practices, educational career, conduct problems, alcohol problems and use of illegal substances was also collected. RESULTS: Young adults who were nicotine-dependent had clearly elevated rates of anxiety, depression and parasuicide. These rates declined after controlling for a previous history of mental health problems and potential confounding factors. After adjustment, nicotine dependence was still associated with anxiety, depression and parasuicide. There was also a significant association with later depression in the group of non-dependent daily smokers. Measures of reduced mental health did not predict later smoking initiation or the development of nicotine dependence. CONCLUSIONS: Mental health was reduced more seriously in nicotine-dependent smokers than in non-dependent smokers. These findings are consistent with the hypothesis that smoking, in particular nicotine dependence, influences mental health. PMID- 19133899 TI - Does sports participation during adolescence prevent later alcohol, tobacco and cannabis use? AB - AIMS: To study whether participation in organized sports during adolescence predicts increased smoking of tobacco, alcohol intoxication and cannabis use from late adolescence to adulthood when controlling for potential confounders. Moreover, to study whether such increased drug use varies according to type of sport (team versus individual), main skills needed (endurance, strength or technical) and level of competition. DESIGN, SETTING AND PARTICIPANTS: Survey of national sample of Norwegian high school students (aged 13-19 years) in 1992 (T1) followed-up in 1994 (T2), 1999 (T3) and 2006 (T4) (n = 3251). MEASUREMENTS: Outcome measures included smoking of tobacco and 12-month prevalences of alcohol intoxication and cannabis use, respectively. Confounders included pubertal timing, friends' drug use, perceived social acceptance, grades and parental socio economic status. FINDINGS: Latent growth curve analyses showed that initial level of participation in organized sports predicted growth in alcohol intoxication. Those involved initially in team sports had greater growth in alcohol intoxication, but lower growth in tobacco use and cannabis use, during the adolescent and early adult years compared to those involved in technical or strength sports. Practising endurance sports, as opposed to technical or strength sports, predicted reduced growth in alcohol intoxication and tobacco use. CONCLUSIONS: Sports participation in adolescence, and participation in team sports in particular, may increase the growth in alcohol intoxication during late adolescent and early adult years, whereas participation in team sports and endurance sports may reduce later increase in tobacco and cannabis use. PMID- 19133900 TI - Sports and drugs: further interpretative hypotheses are necessary. PMID- 19133901 TI - Further comments on the path to drawing reasonable conclusions about prevention. PMID- 19133903 TI - Illicit tobacco: an international problem with an international solution. PMID- 19133904 TI - The absurdity of the use of 'alcohol abuse'. PMID- 19133905 TI - Abortion and substance use: cause or confounding? PMID- 19133908 TI - Introduction to the issue--the alcohol industry and alcohol policy. PMID- 19133909 TI - Alcohol industry and alcohol policy--the challenge ahead. PMID- 19133910 TI - The global alcohol industry: an overview. AB - AIMS: To describe the globalized sector of the alcoholic beverage industry, including its size, principal actors and activities. METHODS: Market research firms and business journalism are the primary sources for information about the global alcohol industry, and are used to profile the size and membership of the three main industry sectors of beer, distilled spirits and wine. FINDINGS: Branded alcoholic beverages are approximately 38% of recorded alcohol consumption world-wide. Producers of these beverages tend to be large multi-national corporations reliant on marketing for their survival. Marketing activities include traditional advertising as well as numerous other activities, such as new product development, product placement and the creation and promotion of social responsibility programs, messages and organizations. CONCLUSIONS: The global alcohol industry is highly concentrated and innovative. There is relatively little public health research evaluating the impact of its many marketing activities. PMID- 19133911 TI - The alcohol industry and trade agreements: a preliminary assessment. AB - AIMS: To review trade agreements, their relation to alcohol control policy and examine the role of the alcohol industry in supporting and attempting to influence trade policy. METHODS: Review of peer review, public health advocacy literature (both pro and con on free trade), business, press and government documents on trade agreements, assess current and potential challenges by trade agreements to alcohol control policy and investigate the means and extent of industry influence in trade agreements. FINDINGS: 'Free' trade agreements reduce trade barriers, increase competition, lower prices and promote alcohol consumption. However, international treaties, negotiated by free trade experts in close consultation with corporate lobbyists and without significant, if any, public health input, governments and corporations contain significant provisions that will result in increased alcohol consumption and may challenge public health measures of other nations as constraints on trade. Conversely, alcohol control measures seek to reduce access and consumption, raise prices and restrict advertising and product promotion. The prospect is for increased alcohol consumption and concomitant problems throughout the world. CONCLUSIONS: Trade agreements challenge effective alcohol control policies. The alcohol industry seeks to influence agreements and can be expected to work through trade agreements to reduce tariffs, increase market access and seek to restrict effective domestic regulations. Further research is needed on the impact of trade agreements and the ongoing role of the industry. Advocates must recognize the inherent conflicts between unbridled free trade and public health, work to exclude alcohol from trade agreements, counter industry influence and protect alcohol control policies. PMID- 19133912 TI - Litigation and alcohol policy: lessons from the US Tobacco Wars. AB - AIMS: This paper explores the role of litigation in preventing alcohol-related harms, identifying lessons from the use of litigation in tobacco control policy in the United States. It analyzes the key components of litigation in an international context, provides a case study of its potential use in addressing the marketing of alcopops to youth and offers recommendations for pursuing litigation strategies in future alcohol policy efforts. METHODS: The paper's analyses are based on both original and secondary legal research. State and federal case law and secondary sources are reviewed in assessing lessons learned from tobacco litigation in the United States and the potential role of litigation in alcohol policy, both in the United States and internationally. Assessment of alcohol litigation cases and state and federal laws and regulations provides the foundation for the alcopops case study. FINDINGS: The tobacco litigation experience demonstrates that litigation is a powerful tool in addressing aggressive marketing by purveyors of addictive products such as alcohol. CONCLUSIONS: To be effective at both national and international levels, litigation should encompass a broad array of legal tactics designed to identify and restrict unfair, deceptive and misleading alcohol marketing tactics and should be utilized in conjunction with complementary prevention strategies. Research conducted on the impact of alcohol marketing on youth alcohol consumption and problems is needed to support potential litigation claims. Developing litigation expertise within the alcohol policy field and building collaboration with litigation specialists in tobacco control should also be considered a high priority. PMID- 19133913 TI - Alcohol research and the alcoholic beverage industry: issues, concerns and conflicts of interest. AB - AIMS: Using terms of justification such as 'corporate social responsibility' and 'partnerships with the public health community', the alcoholic beverage industry (mainly large producers, trade associations and 'social aspects' organizations) funds a variety of scientific activities that involve or overlap with the work of independent scientists. The aim of this paper is to evaluate the ethical, professional and scientific challenges that have emerged from industry involvement in alcohol science. METHOD: Source material came from an extensive review of organizational websites, newspaper articles, journal papers, letters to the editor, editorials, books, book chapters and unpublished documents. RESULTS: Industry involvement in alcohol science was identified in seven areas: (i) sponsorship of research funding organizations; (ii) direct financing of university-based scientists and centers; (iii) studies conducted through contract research organizations; (iv) research conducted by trade organizations and social aspects/public relations organizations; (v) efforts to influence public perceptions of research, research findings and alcohol policies; (vi) publication of scientific documents and support of scientific journals; and (vii) sponsorship of scientific conferences and presentations at conferences. CONCLUSION: While industry involvement in research activities is increasing, it constitutes currently a rather small direct investment in scientific research, one that is unlikely to contribute to alcohol science, lead to scientific breakthroughs or reduce the burden of alcohol-related illness. At best, the scientific activities funded by the alcoholic beverage industry provide financial support and small consulting fees for basic and behavioral scientists engaged in alcohol research; at worst, the industry's scientific activities confuse public discussion of health issues and policy options, raise questions about the objectivity of industry-supported alcohol scientists and provide industry with a convenient way to demonstrate 'corporate responsibility' in its attempts to avoid taxation and regulation. PMID- 19133915 TI - The metabolic syndrome and schizophrenia. AB - OBJECTIVE: To summarize the accumulated data on metabolic syndrome prevalence in patients with schizophrenia, examine evidence for a biological contribution of the mental illness to metabolic risk and review novel options available for management of prediabetic states. METHOD: A Medline search using metabolic syndrome, insulin resistance and insulin sensitivity cross-referenced with schizophrenia was performed on articles published between 1990 and May 2008. RESULTS: Recent evidence indicates that schizophrenia increases predisposition towards metabolic dysfunction independent of environmental exposure. Both fasting and non-fasting triglycerides have emerged as important indicators of cardiometabolic risk, while metformin, thiazolidinediones and GLP-1 modulators may prove promising tools for managing insulin resistance. CONCLUSION: Because of lifestyle, disease and medication effects, schizophrenia patients have significant risk for cardiometabolic disease. Routine monitoring, preferential use of metabolically neutral antipsychotics and lifestyle education are critical to minimizing risk, with a possible role for antidiabetic medications for management of insulin resistant states that do not respond to other treatment strategies. PMID- 19133918 TI - IL15 gene variants are not associated with asthma and atopy. AB - BACKGROUND: Interleukin 15 (IL15) promotes activation and proliferation of CD8+ T cells and enhances the differentiation into Th2 cells. A previous study described five polymorphisms in the IL15 gene to be associated with asthma in a haplotype analysis. AIM: We selected HapMap tagging single nucleotide polymorphisms (SNPs) from IL15 to systematically investigate these IL15 associations in a large population-based sample. METHODS: Genotyping of seven IL15 SNPs was performed using MALDI-TOF MS in a cross-sectional study population of 3099 children from Dresden or Munich (age 9-11 years). All children were phenotyped by standardized and validated protocols for atopic phenotypes. Effects of single SNPs and haplotypes were studied using sas 9.1.3 and haploview. Equivalence tests were performed to prove the significance of negative results. RESULTS: Neither single IL15 polymorphisms nor haplotype analyses showed associations with asthma or atopy after correction for multiple testing. CONCLUSION: These results do not confirm previous case-control studies and suggest that IL15 gene variants do not play an important role in the development for asthma or other atopic disorders. PMID- 19133919 TI - Serum eosinophil granule proteins predict asthma risk in allergic rhinitis. AB - BACKGROUND: Allergic rhinitis is a common disease, in which some patients will deteriorate or develop asthma. It is important to characterize these patients, thereby offering the possibility for prevention. This study evaluated eosinophil parameters as potential indicators of deteriorating allergic airway disease. METHODS: The subjects of the study included all patients who suffered seasonal allergic rhinitis and had participated in a study 6 years earlier, in which blood eosinophils, serum eosinophil cationic protein (ECP) serum eosinophil peroxidase (EPO), nasal lavage ECP and nasal lavage EPO levels were measured. Patients in the present study were interviewed on occurrence of rhinitis symptoms during the last season, rhinitis outside season, asthma-like symptoms and asthma diagnosis, and were skin-prick tested for common aeroallergens. Eosinophil parameters from the study 6 years earlier were then tested for the ability to predict occurrence of new allergies, worsening of rhinitis and occurrence of asthma. RESULTS: Forty four patients participated in the study. In four patients seasonal rhinitis symptoms had deteriorated, 10 had experienced perennial rhinitis symptoms, 14 reported asthma-like symptoms and seven had been diagnosed with asthma. Thirteen had developed additional sensitization. Patients developing asthma-like symptoms compared with patients with no such symptoms had significantly higher serum ECP (16.7 microg/l vs 8.2 microg/l; P < or = 0.01) and serum EPO (17.9 microg/l vs 8.8 microg/l; P < or = 0.05). Results were similar, considering patients diagnosed with asthma. Blood eosinophils and nasal lavage parameters were not related to development of asthma and asthma-like symptoms. No eosinophil parameter was related to deterioration of rhinitis or additional sensitization. CONCLUSION: Serum ECP and EPO in patients with seasonal rhinitis demonstrated a high predictive ability for later development of asthma. PMID- 19133920 TI - How to prescribe antihistamines for chronic idiopathic urticaria: desloratadine daily vs PRN and quality of life. AB - BACKGROUND: Chronic idiopathic urticaria (CIU) impairs quality of life (QoL). Currently, no consensus exists regarding how second-generation H(1) antihistamines (proven to control CIU symptoms) should be taken long-term: as daily treatment or only when symptoms return (PRN). We sought to determine which regimen improves or better maintains QoL in CIU: desloratadine (DL) daily or PRN. METHODS: Subjects with CIU initially responding to DL 5 mg/day for 4 weeks were randomized for an additional 8 weeks, to DL 5 mg/day (arm 1: 'continuous', n = 46) or to DL only on days when urticarial wheals were present (arm 2: "PRN", n = 60). To ensure blinding, treatment was presented in both arms as a combination of daily treatment (arm 1: DL; arm 2: placebo), plus a "rescue" tablet (arm 1: placebo; arm 2: DL) to be taken only in case of symptoms. The main outcome measure was QoL assessed by the VQ-Dermato, a validated French QoL instrument, and the Dermatology Life Quality Index (DLQI). RESULTS: At 4 and 8 weeks after randomization, subjects taking continuous DL showed statistically significant improvements in VQ-Dermato Global Index score (P = 0.001 and P = 0.016, respectively) and dimension scores for daily living activity, mood state, and social functioning vs subjects taking DL PRN. Improvement in DLQI score at Week 4 was also significantly greater with continuous DL (P = 0.001). CONCLUSION: Continuous daily therapy with DL 5 mg is a better regimen than PRN treatment to maintain or improve QoL in subjects with CIU. PMID- 19133921 TI - Genetic variation in ORM1-like 3 (ORMDL3) and gasdermin-like (GSDML) and childhood asthma. AB - BACKGROUND: A genome-wide association study identified ORM1-like 3 (orosomucoid 1 like 3, ORMDL3) as an asthma candidate gene. Single nucleotide polymorphisms (SNPs) in the region including ORMDL3 on chromosome 17q21 were related to childhood asthma risk and ORMDL3 expression levels in Europeans. OBJECTIVE: We examined whether polymorphisms in ORMDL3 and the adjacent gasdermin-like (GSDML) gene associated with asthma in the genome-wide association study are related to childhood asthma and atopy in a Mexico City population. METHODS: We genotyped rs4378650 in ORMDL3 and rs7216389 in GSDML in 615 nuclear families consisting of asthmatic children aged 4-17 years and their parents. Atopy was determined by skin prick tests to 25 aeroallergens. RESULTS: Individuals carrying the C allele of rs4378650 or the T allele of rs7216389 had increased risk of asthma [relative risk (RR) = 1.73, 95% confidence interval (CI) 1.19-2.53, P = 0.003 for one or two copies of rs4378650 C, and RR = 1.64, 95% CI 1.12-2.38, P = 0.009 for one or two copies of rs7216389 T). Linkage disequilibrium between the two SNPs was high (r(2) = 0.92). Neither of the SNPs was associated with the degree of atopy. A meta-analysis of five published studies on rs7216389 in nine populations gave an odds ratio for asthma of 1.44 (95% CI, 1.35-1.54, P < 0.00001). CONCLUSIONS: Our results and the meta-analysis provide evidence to confirm the finding from a recent genome-wide association study that polymorphisms in ORMDL3 and the adjacent GSDML may contribute to childhood asthma. PMID- 19133923 TI - Update on the evaluation of hypersensitivity reactions to betalactams. AB - Hypersensitivity reactions to betalactams (BLs) are classified as immediate or nonimmediate. The former usually appear within 1 h of drug-intake and are mediated by specific IgE-antibodies. Nonimmediate reactions are those occurring more than 1 h after drug-intake, and they can be T-cell mediated. The diagnostic evaluation of allergic reactions to BLs has changed over the last 5 years, for several reasons. Major and minor determinants are no longer commercially available for skin testing in many countries. In immediate allergic reactions, the sensitivity of skin testing and immunoassays is decreasing and new in vitro methods, such as the basophil activation test, are gaining importance for diagnosis. For nonimmediate reactions, skin testing appears to be less sensitive than previous results, although more studies need to be carried out in this direction. Nevertheless, the drug provocation test is still necessary for diagnosis. PMID- 19133917 TI - The allergic sensitization in infants with atopic eczema from different countries. AB - BACKGROUND: No study has compared allergic sensitization patterns in infants with atopic eczema from different countries. The aim of this study was to investigate the patterns of allergic sensitization in a cohort of infants with atopic eczema participating in a multicentre, international study. METHODS: Two thousand one hundred and eighty-four infants (mean age 17.6 months) with atopic eczema from allergic families were screened in 94 centres in 12 countries to participate in a randomized trial for the early prevention of asthma. Clinical history, Severity Scoring of Atopic Dermatitis Index, measurements for total serum IgE and specific IgE antibodies to eight food and inhalant allergens were entered into a database before randomization to treatment. A history of type of feeding in the first weeks of life and exposure to animals was recorded. RESULTS: A total of 52.9% of the infants had raised total IgE, and 55.5% were sensitized to at least one allergen. There was a wide difference in the total IgE values and in the sensitization rates to foods and aeroallergens among infants from different countries. The highest prevalence rates of allergen-sensitized infants were found in Australia (83%), the UK (79%) and Italy (76%). Infants from Belgium and Poland consistently had the lowest sensitization rates. In each country, a characteristic pattern of sensitization was found for aeroallergens (house dust mite > cat > grass pollen > Alternaria), but not for food allergens. CONCLUSIONS: In infants with atopic eczema, there is a wide variation in the pattern of allergic sensitization between countries, and data from one country are not necessarily generalizable to other countries. PMID- 19133924 TI - Benzylpenicillin skin testing is still important in diagnosing immediate hypersensitivity reactions to penicillins. AB - BACKGROUND: The fact that both Hollister-Stier and Allergopharma ceased the production of penicilloyl-polylysine (PPL) and minor determinant mixture (MDM) in 2004 is severely hampering the diagnosis of beta-lactam hypersensitivity and may produce negative consequences. OBJECTIVE: To assess the contribution of skin testing with benzylpenicillin to the diagnosis of immunoglobulin E-mediated hypersensitivity to penicillins, in order to determine how much such testing could compensate for PPL and MDM unavailability. METHODS: We selected patients with histories of immediate reactions to penicillins and positive results to skin tests for one or more penicillin reagents (PPL, MDM, or benzylpenicillin), one or more semi-synthetic penicillins (ampicillin, amoxicillin, or piperacillin), or both. RESULTS: A total of 300 patients were selected, 105 in the French center and 195 in the Italian centers. Amoxicillin and ampicillin were the main responsible drugs. The most common clinical manifestation was anaphylaxis. The reagents most frequently positive to skin tests were amoxicillin (188, 62.7%), ampicillin (151, 50.3%), and benzylpenicillin (111, 37.0%). Among the 300 subjects, 113 (37.7%) were positive only to semi-synthetic penicillins, 109 (36.3%) to both semi-synthetic penicillins and the classic penicillin reagents, and 78 (26.0%) only to the latter. In the last group, 64 (21.3% of the 300 subjects) were positive only to PPL and/or MDM and 14 (4.7%) to benzylpenicillin, of whom 8 (2.7%) were positive only to the latter. CONCLUSIONS: Skin testing with benzylpenicillin can partially compensate for PPL and MDM unavailability. Moreover, it can slightly increase the allergologic workup's sensitivity and therefore reduce the number of potentially dangerous challenges. PMID- 19133922 TI - Cornulin, a marker of late epidermal differentiation, is down-regulated in eczema. AB - BACKGROUND: Eczema is a common chronic inflammatory skin disorder which shows strong genetic predisposition. To identify new potential molecular determinants of the disease pathogenesis, we performed a gene expression study in an eczema mouse model. This analysis identified a marked down regulation of the cornulin gene (CRNN), a member of the epidermal differentiation complex, in the eczema like skin. We then investigated CRNN as an eczema candidate gene and studied its polymorphism and the expression in the skin of eczema patients. METHODS: An eczema-like phenotype was induced in mice by allergen (Der p2) patching. Gene expression analysis was performed with the subtractive suppression hybridization method and validated by real time PCR and the transmission disequilibrium test was used to test for genetic associations in 406 multiplex eczema families. RESULTS: Der p 2 patched mice developed a localized eczema and a Th 2 skewed systemic response. Real time PCR analysis confirmed a down regulation of CRNN mRNA in eczema-like skin in the mouse model and in human eczema. The CRNN polymorphism rs941934 was significantly associated with atopic eczema in the genetic analysis (P = 0.006), though only as part of an extended haplotype including a known associated variant (2282del4) in the filaggrin gene. CONCLUSIONS: CRNN mRNA expression is decreased in eczematous skin. Further studies are needed to verify whether the associated cornulin polymorphism contribute to the genetic susceptibility in eczema. PMID- 19133925 TI - News and issues that affect organ and tissue transplantation. AB - Australia, Canada and the United Kingdom launch national programs. PMID- 19133926 TI - Transplantation oversight--finding the right balance between the layperson, the transplant professional and the regulator. PMID- 19133927 TI - Cardiac allograft hypertrophy: a new target for therapy, a surrogate marker for survival? PMID- 19133928 TI - Allocation or rationing--word choice is crucial. PMID- 19133929 TI - Nine things you might not say or hear in transplantation. PMID- 19133930 TI - Nonadherence consensus conference summary report. AB - This report is a summary of a 'Consensus Conference' on nonadherence (NA) to immunosuppressants. Its aims were: (1) to discuss the state-of-the-art on the definition, prevalence and measurement of NA, its risk factors and impact on clinical and economical outcomes and interventions and (2) to provide recommendations for future studies. A two-day meeting was held in Florida in January 2008, inviting 66 medical and allied health adherence transplant and nontransplant experts. A scientific committee prepared the meeting. Consensus was reached using plenary and interactive presentations and discussions in small break-out groups. Plenary presenters prepared a summary beforehand. Break-out group leaders initiated discussion between the group members prior to the meeting using conference calls and e-mail and provided a summary afterward. Conclusions were that NA: (a) is more prevalent than we assume; (b) is hard to measure accurately; (c) tends to confer worse outcomes; (d) happens for a number of reasons, and system-related factors including the patient's culture, the healthcare provider and the setting and (e) it is not currently known how to improve adherence. This consensus report provided some roadmaps for future studies on this complicated, multifaceted problem. PMID- 19133931 TI - Long-term survival of nonhuman primate islets implanted in an omental pouch on a biodegradable scaffold. AB - The aim of this study was to test whether an omental pouch can be used as an alternative site for islet implantation in diabetic monkeys. Here we report the successful engraftment of islets in diabetic cynomolgus monkeys when loaded on a synthetic biodegradable scaffold and placed in an omental pouch. One autologous and five allogeneic diabetic monkey transplants under the cover of steroid-free immune suppression (SFIS) were undertaken. Fasting blood glucose (FBG) and C peptide (CP), exogenous insulin requirements (EIR), intravenous glucose tolerance test (IVGTT), A1C and histopathology were used to assess islet engraftment and survival. All animals achieved CP levels > 1.0 ng/mL following transplant, a 66 92% posttransplant decrease in EIR and reduced A1C. Following graft removal, CP became negative and histopathological analysis of the explanted grafts demonstrated well-granulated and well-vascularized, insulin-positive islets, surrounded by T-cell subsets and macrophages. Compared to intrahepatic allogeneic islet transplants (n = 20), there was a delayed engraftment for omental pouch recipients but similar levels of CP production were ultimately achieved, with a broad range of IEQ/kg transplanted in both sites. Our results suggest this extrahepatic transplantation site has potential as an alternative site for clinical islet cell transplantation. PMID- 19133932 TI - Effects of donor age and cell senescence on kidney allograft survival. AB - The biological processes responsible for somatic cell senescence contribute to organ aging and progression of chronic diseases, and this may contribute to kidney transplant outcomes. We examined the effect of pre-existing donor aging on the performance of kidney transplants, comparing mouse kidney isografts and allografts from old versus young donors. Before transplantation, old kidneys were histologically normal, but displayed an increased expression of senescence marker p16(INK4a). Old allografts at day 7 showed a more rapid emergence of epithelial changes and a further increase in the expression of p16(INK4a). Similar but much milder changes occurred in old isografts. These changes were absent in young allografts at day 7, but emerged by day 21. The expression of p16(INK4a) remained low in young kidney allografts at day 7, but increased with severe rejection at day 21. Isografts from young donors showed no epithelial changes and no increase in p16(INK4a). The measurements of the alloimmune response-infiltrate, cytology, expression of perforin, granzyme B, IFN-gamma and MHC-were not increased in old allografts. Thus, old donor kidneys display abnormal parenchymal susceptibility to transplant stresses and enhanced induction of senescence marker p16(INK4a), but were not more immunogenic. These data are compatible with a key role of somatic cell senescence mechanisms in kidney transplant outcomes by contributing to donor aging, being accelerated by transplant stresses, and imposing limits on the capacity of the tissue to proliferate. PMID- 19133933 TI - Coagulation and fractionated plasma separation and adsorption. PMID- 19133934 TI - Incidence of JC-virus replication after rituximab therapy in solid-organ transplant patients. PMID- 19133935 TI - Haplotype sharing excludes orthologous COL4A3, COL4A4 or MYH9 loci in hereditary nephritis in bull terriers. PMID- 19133936 TI - Variation in ovine CAPN3 is not associated with meat tenderness. PMID- 19133937 TI - Quantitative trait loci for fatty acid composition in longissimus dorsi and abdominal fat: results from a White Duroc x Erhualian intercross F2 population. AB - A whole-genome scan was performed on 660 F(2) animals including 250 barrows and 410 gilts in a White Duroc x Erhualian intercross population to detect quantitative trait loci (QTL) for fatty acid composition in the longissimus dorsi muscle and abdominal fat. A total of 153 QTL including 63 genome-wide significant QTL and 90 suggestive effects were identified for the traits measured. Significant effects were mainly evident on pig chromosomes (SSC) 4, 7, 8 and X. No association was detected on SSC3 and 11. In general, the QTL detected in this study showed distinct effects on fatty acid composition in the longissimus muscle and abdominal fat. The QTL for fatty acid composition in abdominal fat did not correspond to those identified previously in backfat and the majority of QTL for the muscle fatty acid composition were mapped to chromosomal regions different from previous studies. Two regions on SSC4 and SSC7 showed significant pleiotropic effects on monounsaturated (MUFA) and polyunsaturated fatty acid (PUFA) in both longissimus muscle and abdominal fat. Another two QTL with significant multi-faceted effects on MUFA and PUFA in the longissimus muscle were found each on SSC8 and SSCX. Chinese Erhualian alleles were associated with increased ratios of MUFA to saturated fatty acid at most of the QTL detected, showing beneficial effect in terms of human health. PMID- 19133938 TI - Mapping and expression analyses during porcine foetal muscle development of 12 genes involved in histone modifications. AB - Histone modifications (methylation and demethylation) regulate gene expression and play a role in cell proliferation and differentiation by their actions on chromatin structure. In this context, we studied the temporal expression profiles of genes acting on histone methylation and demethylation during skeletal muscle proliferation and differentiation. Quantitative real-time PCR was used to quantify the mRNA levels of CARM1, JARID1A, JMJD2A, LSD1, PRMT2, PRMT5, SMYD1, SMYD2, SMYD3, SETDB1, Suv39h2 and SUZ12 in foetal skeletal muscle. Our results showed that CARM1, JARID1A, JMJD2A, SMYD1 and SMYD2 were differentially expressed in embryonic muscles of 33 days post-conception (dpc), 65 dpc and 90 dpc. These 12 genes were mapped to porcine chromosomes (SSC) 2q21-24, 5q25, 6q35, 6q12-21, 6p15, 7q21, 3q21-27, 9q26, 10p16, 4q15-16, 10q14-16 and 12p12 respectively. Taking into account the reported QTL mapping results, gene expression analysis and radiation hybrid mapping results, these results suggest that five genes (CARM1, JARID1A, JMJD2A, SMYD1 and SMYD2) could be good candidate genes for growth and backfat thickness traits. PMID- 19133939 TI - Association of single nucleotide polymorphisms in the endothelial differentiation sphingolipid G-protein-coupled receptor 1 gene with marbling in Japanese Black beef cattle. AB - Marbling defined by the amount and distribution of intramuscular fat, so-called Shimofuri, is an economically important trait of beef cattle in Japan. The endothelial differentiation sphingolipid G-protein-coupled receptor 1 (EDG1) gene, involved in blood vessel formation, has been previously shown to be expressed at different levels in musculus longissimus muscle between low-marbled and high-marbled steer groups. It is located within the genomic region of a quantitative trait locus for marbling, and thus was considered as a positionally functional candidate for the gene responsible for marbling. In this study, two single nucleotide polymorphisms (SNPs) in the 5' untranslated region (UTR) and the 3' UTR of EDG1, referred to as c.-312A>G and c.*446G>A, respectively, were detected between the two steer groups. The two SNPs were associated with the predicted breeding value for beef marbling standard number by analyses using a population of Japanese Black beef cattle. The effect of genotypes at each of the SNPs on the predicted breeding value for subcutaneous fat thickness was not statistically significant (P > 0.05). Reporter gene assays revealed no significant differences in gene expression between alleles at each of the SNPs. These findings suggest that EDG1 SNPs, although they may not be regarded as a causal mutation, may be useful for effective marker-assisted selection to increase the levels of marbling in Japanese Black beef cattle. PMID- 19133940 TI - Inbreeding coefficients for X-linked and autosomal genes in consanguineous marriages in Spanish populations: the case of Guipuzcoa (Basque Country). AB - Inbreeding patterns over the past two centuries have been studied more extensively in Spain and Italy than anywhere else in Europe. Consanguinity studies in mainland Spain have shown that populations settled along the Cantabrian cornice share inbreeding patterns that distinguish them from other populations further south. A visual representation of spatial variations of two key inbreeding variables is presented here for the first time via contour maps. This paper also analyzes time trends of mean inbreeding coefficients for X-linked (F(x)) and autosomal genes (F) (1862-1995) together with variations in F(x)/F ratios in Guipuzcoa, the most autochthonous Spanish Basque province. Because close cousin marriages are a mark of identity of the study population, we evaluated the contribution of uncle-niece/aunt-nephew (M12) and first cousin (M22) marriages to F(x) and F values and compared the frequencies of M12 and M22 pedigree subtypes and their corresponding F(x)/F ratios to those found in other Spanish populations. The mean Fx and F inbreeding levels in Guipuzcoa for the 134 year period analyzed were 1.51 x 10(-3) and 1.04 x 10(-3), respectively, and the F(x)/F ratio was seen to be very stable over time. Our findings show that major similarities exist for close consanguineous marriage subtypes between Basque and non-Basque Spanish populations, despite significant geographic variability in terms of first cousin pedigrees. The distortion seems to be caused by Guipuzcoa. The F(x)/F ratios for first cousins in Spanish populations were higher than expected (1.25), with values ranging from 1.34 to 1.48. The findings of the present study may be useful for advancing knowledge on the effects of the interaction between biology and culture and for exploring associations between mating patterns and the prevalence of certain diseases. PMID- 19133941 TI - Distinct clinical characteristics of tuberous sclerosis complex patients with no mutation identified. AB - Tuberous Sclerosis Complex (TSC) is a multi-system disorder that is highly variable in its clinical presentation. Current molecular diagnostic methods permit identification of mutations in either TSC1 or TSC2 in 75-85% of TSC patients. Here we examine the clinical characteristics of those TSC patients who have no mutation identified (NMI). A retrospective review of our patient population that had comprehensive testing for mutations in TSC1/TSC2 identified 23/157 (15%) that were NMI. NMI patients had a lower incidence of brain findings on imaging studies, neurological features, and renal findings than those with TSC2 mutations. In contrast, NMI patients had a lower incidence of seizures than TSC patients with TSC1 mutations, but had a higher incidence of both renal angiomyolipomas and pulmonary lymphangioleiomyomatosis. This distinct constellation of findings suggest that NMI patients may have a unique molecular pathogenesis, different from that seen in TSC patients with the usual mutations in TSC1 and TSC2. We suggest that the mechanisms of disease in these patients include both mosaicism for a TSC2 mutation, and unusual non-coding region mutations in TSC2. PMID- 19133942 TI - Heterogeneous disease modeling for Hardy-Weinberg disequilibrium in case-control studies: application to renal stones and calcium-sensing receptor polymorphisms. AB - Renal stone formation due to hypercalciuria is a relatively common disorder with clear evidence for genetic predisposition, but cryptic phenotypic heterogeneity has hampered identification of candidate genes. The R990G single-nucleotide polymorphism (SNP) of the calcium sensing receptor (CASR) gene has been associated with hypercalciuria in stone formers and shows the appropriate functional phenotype in cell culture. In our preliminary association analysis of a case-control cohort, however, we observed significant Hardy-Weinberg disequilibrium (HWD) for the cases (n= 223), but not controls (n= 676) at the R990G locus, pointing us toward the general disease model incorporating HWD. Because there is an adjacent CASR SNP, A986S, which is in negative linkage disequilibrium with R990G, we extended the general disease model to enable testing of a two-site hypothesis. In our data set, there is no lack of fit (P= .345) for the single-locus model for the R990G genotype, and likelihood ratio testing favors a recessive effect with an eight-fold increase in risk (P < .001) for GG homozygotes, relative to wild-type, based on a population prevalence of 2%. Addition of the A986S genotype provides no additional information either by itself or when included in our two-site model. PMID- 19133943 TI - Dental research--it's alive and well and relevant. PMID- 19133944 TI - The role of alcohol in oral carcinogenesis with particular reference to alcohol containing mouthwashes. AB - Worldwide, oral cancer represents approximately 5 per cent of all malignant lesions, with over 800 new intra-oral squamous cell carcinomas registered in Australia each year. Despite recent advances in therapy, the five-year survival rate remains around 50 per cent and the sequelae of treatment can be seriously debilitating. It has been long established that smoking and alcohol consumption are risk factors linked to the development of oral cancer. This review assesses the epidemiological evidence, supportive in vitro studies and mechanism by which alcohol is involved in the development of oral cancer. Further, we review the literature that associates alcohol-containing mouthwashes and oral cancer. On the basis of this review, we believe that there is now sufficient evidence to accept the proposition that alcohol-containing mouthwashes contribute to the increased risk of development of oral cancer and further feel that it is inadvisable for oral healthcare professionals to recommend the long-term use of alcohol containing mouthwashes. PMID- 19133945 TI - The Monitor Practice Programme: is non-invasive management of dental caries in private practice effective? AB - BACKGROUND: This paper reviews the efficacy of an intensive, preventive-based, non-invasive approach to the management of dental caries within a randomized controlled trial. METHODS: The primary efficacy measure was the two-year DMFS increment. Changes in risk status, fluoride history, number of emergency visits and toothaches, along with demographic variables such as age, gender, health problems, and the location of the dental practice attended were measured. Regression analysis was undertaken to adjust for potential confounding variables. RESULTS: Nine hundred and two patients were recruited within 22 dental practices between May 2005 and March 2006. Baseline DMFS did not differ significantly between the control and study groups (p = 0.83). Age (p < 0.001), health status (p = 0.005), baseline risk (p < 0.001) and fluoride history (p < 0.001) were all independent significant predictors of two-year DMFS increment. Gender approached significance (p = 0.08). There were no statistically significant differences between the groups in the incidence of toothaches (p = 0.1) or number of treatment visits required (p = 0.35). There was a significant difference in the two-year incremental DMFS score in the study group compared to the control group (mean difference 2.2; p < 0.001). After adjusting for confounding variables the difference in the DMFS increment between the control and study groups remained significant (mean difference 1.7; p < 0.001). CONCLUSIONS: The results indicate efficacy of the preventive programme. Efficacy was independent of age, gender, medical concerns, fluoride history, or previous history of dental caries, in a population of patients attending for treatment in private dental practices, in a variety of locations, on a relatively short-term basis (two years). While encouraging, it will be essential that these results are followed over a longer period of time in order to determine whether the benefits are maintained. PMID- 19133946 TI - Protective effect of yogurt extract on dental enamel demineralization in vitro. AB - BACKGROUND: Casein phosphopeptides (CPPs) are phosphorylated casein-derived peptides produced synthetically by proteolytic digestion of alpha(s1)-, alpha(s2) and beta-casein. The anticariogenic activity of CPPs is due to their ability to stabilize high levels of amorphous calcium phosphate (ACP) on tooth surface, preventing demineralization and enhancing remineralization of enamel caries. The aim of this study was to test the in vitro ability of natural CPPs (contained in yogurt) to prevent demineralization and promote remineralization of dental enamel. METHODS: Eighty human molars were used. After standardizing an in vitro demineralization procedure for producing artificial caries (Group 1: pH 4.8; Group 2: pH 3.97), this procedure was used on teeth, but with the addition of natural CPPs (Group 3: pH 4.8; Group 4: pH 3.97). The effects of these procedures were evaluated by quantitative analysis (change in weight and calcium titration) and qualitative analysis (SEM). Statistical analysis of the results was performed using ANOVA. RESULTS: Statistical analysis showed significant differences in weight changes between the groups with and without natural CPPs. The SEM observation showed the protective effects of natural CPPs. CONCLUSIONS: The results demonstrated that CPPs contained in yogurt have an inhibitory effect on demineralization and promote the remineralization of dental enamel. PMID- 19133947 TI - The incidence of mandibular third molar impactions in different skeletal face types. AB - BACKGROUND: The cause of mandibular third molar impaction is said to be due to inadequate space between the distal of the second mandibular molar and the anterior border of the ascending ramus of the mandible. The amount of space is determined primarily by facial growth. This study aimed to assess whether different patterns of facial growth lead to a different incidence of mandibular third molar impaction. It was hypothesized that those with predominantly horizontal (brachyfacial) would have lower incidence of mandibular third molar impaction compared with those with a predominantly vertical growth pattern (dolichofacial). METHODS: Ninety-eight dental records were sourced from the records of orthodontic patients in the Royal Dental Hospital of Melbourne and all lateral cephalometric radiographs and orthopantomograms were assessed. The degree of impaction was determined by the Pell and Gregory system, and the facial type categorized by the facial axis angle. RESULTS: The overall rate of mandibular third molar impaction was 58.76 per cent. Those with a facial axis angle >93 (brachyfacials) demonstrated an almost two times lower incidence of mandibular third molar impaction as compared to subjects with a facial axis angle <87 (dolichofacials). CONCLUSIONS: Within the limitations of the study, it was concluded that the greater horizontal facial growth pattern of brachyfacial subjects over dolichofacial subjects provides increased space for full eruption of the mandibular third molars. PMID- 19133948 TI - Dentine bond strength and microleakage of flowable composite, compomer and glass ionomer cement. AB - BACKGROUND: To assess in vitro the dentine bond strength and microleakage of three Class V restorations viz. flowable composite, compomer and glass ionomer cement. METHODS: Eighteen dentine specimens were prepared and randomly distributed among three groups. Three kinds of restoration materials were each bonded on prepared dentine surfaces in three groups as per the manufacturers' instructions. Group Aelite: Tyrian SPE (a no-rinse, self-priming etchant) + One Step Plus (an universal dental adhesive) + Aeliteflo (a flowable composite); Group Dyract: Prime & Bond NT (a no-rinse, self-priming dental adhesive) + Dyract AP (a compomer); Group GlasIonomer: GlasIonomer Type II (a self-cured restorative glass ionomer). Fifteen dentine/restoration microtensile bond test specimens were prepared from each group and were subjected to microtensile bond strength testing. The bond interfaces were observed morphologically using a scanning electron microscope (SEM). Twenty-four cervical cavities of 4.0 mm mesiodistal length, 2.0 mm occlusogingival height and 1.5 mm depth were prepared at the cemento-enamel junction (CEJ) on both buccal and lingual surfaces of each tooth. The cavities were each filled with flowable composite (Group Aelite), compomer (Group Dyract) and glass ionomer cement (Group GlasIonomer) using the same material and methods as for the microtensile bond tests. Microleakage of each restoration was evaluated by the ratio of the length of methylene blue penetration along the tooth-restoration interface and the total length of the dentine cavity wall on the cut surface. RESULTS: One-way ANOVA and least significant difference (LSD) tests revealed statistically significant differences among the dentine bond strength for Group Aelite (28.4 MPa), Group Dyract (15.1 MPa) and Group GlasIonomer (2.5 MPa). SEM images showed intimate adaptation in the restoration/dentine interfaces of Group Aelite and Group Dyract. All of the systems tested in this study presented microleakage. However, both Group Aelite (0.808) and Group Dyract (0.863) had significantly less microleakage than Group GlasIonomer (0.964). There were no statistically significant microleakage differences between Group Aelite and Group Dyract, and no statistically significant microleakage differences between the occlusal margin and gingival margin. CONCLUSIONS: None of the systems tested in this study completely eliminated microleakage. However, both the flowable composite and compomer provided stronger dentine bond strengths and better margin sealing than the conventional glass ionomer cement. Occlusal forces exerted the same effects on microleakage of the occlusal margin and gingival margin in cervical cavities. PMID- 19133949 TI - Retention of cast crown copings cemented to implant abutments. AB - BACKGROUND: The cementation of crowns to dental implant abutments is an accepted form of crown retention that requires consideration of the properties of available cements within the applied clinical context. Dental luting agents are exposed to a number of stressors that may reduce crown retention in vivo, not the least of which is occlusal loading. This study investigated the influence of compressive cyclic loading on the physical retention of cast crown copings cemented to implant abutments. METHODS: Cast crown copings were cemented to Straumann synOcta titanium implant abutments with three different readily used and available cements. Specimens were placed in a humidifier, thermocycled and subjected to one of four quantities of compressive cyclic loading. The uniaxial tensile force required to remove the cast crown copings was then recorded. RESULTS: The mean retention values for crown copings cemented with Panavia-F cement were statistically significantly greater than both KetacCem and TempBond non-eugenol cements at each compressive cyclic loading quantity. KetacCem and TempBond non-eugenol cements produced relatively low mean retention values that were not statistically significantly different at each quantity of compressive cyclic loading. Compressive cyclic loading had a statistically significant effect on Panavia-F specimens alone, but increased loading quantities produced no further statistically significant difference in mean retention. CONCLUSIONS: Within the limitations of the current in vitro conditions employed in this study, the retention of cast crown copings cemented to Straumann synOcta implant abutments with a resin, glass ionomer and temporary cement was significantly affected by cement type but not compressive cyclic loading. Resin cement is the cement of choice for the definitive non-retrievable cementation of cast crown copings to Straumann synOcta implant abutments out of the three cements tested. PMID- 19133950 TI - Caries risk reduction in patients attending a caries management clinic. AB - BACKGROUND: A Caries Management Clinic was established for patients at high risk of caries aiming to reduce caries incidence to close to zero. That is, to prevent new lesions on existing sound surfaces, along the susceptible restoration margins, and to remineralize existing cavitated and non-cavitated lesions. Twenty patients attended the clinic every two weeks from April to December 2005. METHODS: The Caries Management System is a ten-step non-invasive strategy to arrest and remineralize early lesions and includes consideration of the patient at risk, the status of each individual lesion, patient management, clinical management, and monitoring. RESULTS: After six months, there was a 42 per cent increase in gingival sites having Gingival Index scores of zero, and a 21 per cent decrease in sites having Gingival Index scores of 2 compared to baseline (chi(2 )=( )137.67, 4 df, p = 0.00001). Ready to change (RTC) patients had significantly fewer sites scored GI 0.05). CONCLUSIONS: This preliminary study has shown that the infusion of ozone into non carious dentine prevented biofilm formation in vitro from S. mutans and L. acidophilus over a four-week period. The possibility exists that ozone treatment may alter the surface wettability of dentine through reaction with organic constituents. PMID- 19133952 TI - The oral health status of patients on oral bisphosphonates for osteoporosis. AB - BACKGROUND: The oral health status of patients on bisphosphonates is the key to the patient's ongoing health and well-being. If they are orally healthy, invasive bone procedures, particularly extractions can be avoided, then the risk of osteonecrosis of the jaws (ONJ) is low. METHODS: The records of 49 consecutive patients on oral bisphosphonates, referred to the Oral and Maxillofacial Surgery Unit (OMSU) for an oral health check and probable extractions, were retrospectively reviewed. The DMFT, periodontal and pathologic state were calculated from the OPG radiographs. An age and gender matched control group, from patients referred to the OMSU but who were not on oral bisphosphonates, were similarly assessed. Community data were also obtained. RESULTS: The DMFT score for the oral bisphosphonate group was 29: Decayed 3, Missing 10, Filled 16. The control group DMFT score was 24: Decayed 5, Missing 11, Filled 8. Both groups had advanced periodontal disease (over 95 per cent) and were medically compromised (over 90 per cent). The DMFT for general community data for age matched government pensioners was 19.1: Decayed 0.8, Missing 10.4, Filled 7.9. With severe periodontal disease 23 per cent. Thus, the oral health of the oral bisphosphonate group was similar to the control group and both had more decayed teeth and periodontal disease than community values. CONCLUSIONS: This study confirms that one cannot assume that a patient on an oral bisphosphonate for osteoporosis has a healthy mouth. It supports the view that all patients on bisphosphonates need to be seen by a dentist either before or soon after commencement of bisphosphonate therapy. PMID- 19133953 TI - Factors associated with infrequent dental attendance in the Australian population. PMID- 19133958 TI - Microleakage of composite resin restorations. PMID- 19133959 TI - Control of cell volume in skeletal muscle. AB - Regulation of cell volume is a fundamental property of all animal cells and is of particular importance in skeletal muscle where exercise is associated with a wide range of cellular changes that would be expected to influence cell volume. These complex electrical, metabolic and osmotic changes, however, make rigorous study of the consequences of individual factors on muscle volume difficult despite their likely importance during exercise. Recent charge-difference modelling of cell volume distinguishes three major aspects to processes underlying cell volume control: (i) determination by intracellular impermeant solute; (ii) maintenance by metabolically dependent processes directly balancing passive solute and water fluxes that would otherwise cause cell swelling under the influence of intracellular membrane-impermeant solutes; and (iii) volume regulation often involving reversible short-term transmembrane solute transport processes correcting cell volumes towards their normal baselines in response to imposed discrete perturbations. This review covers, in turn, the main predictions from such quantitative analysis and the experimental consequences of comparable alterations in extracellular pH, lactate concentration, membrane potential and extracellular tonicity. The effects of such alterations in the extracellular environment in resting amphibian muscles are then used to reproduce the intracellular changes that occur in each case in exercising muscle. The relative contributions of these various factors to the control of cell volume in resting and exercising skeletal muscle are thus described. PMID- 19133960 TI - Testing co-evolutionary hypotheses over geological timescales: interactions between Mesozoic non-avian dinosaurs and cycads. AB - The significance of co-evolution over ecological timescales is well established, yet it remains unclear to what extent co-evolutionary processes contribute to driving large-scale evolutionary and ecological changes over geological timescales. Some of the most intriguing and pervasive long-term co-evolutionary hypotheses relate to proposed interactions between herbivorous non-avian dinosaurs and Mesozoic plants, including cycads. Dinosaurs have been proposed as key dispersers of cycad seeds during the Mesozoic, and temporal variation in cycad diversity and abundance has been linked to dinosaur faunal changes. Here we assess the evidence for proposed hypotheses of trophic and evolutionary interactions between these two groups using diversity analyses, a new database of Cretaceous dinosaur and plant co-occurrence data, and a geographical information system (GIS) as a visualisation tool. Phylogenetic evidence suggests that the origins of several key biological properties of cycads (e.g. toxins, bright coloured seeds) likely predated the origin of dinosaurs. Direct evidence of dinosaur-cycad interactions is lacking, but evidence from extant ecosystems suggests that dinosaurs may plausibly have acted as seed dispersers for cycads, although it is likely that other vertebrate groups (e.g. birds, early mammals) also played a role. Although the Late Triassic radiations of dinosaurs and cycads appear to have been approximately contemporaneous, few significant changes in dinosaur faunas coincide with the late Early Cretaceous cycad decline. No significant spatiotemporal associations between particular dinosaur groups and cycads can be identified - GIS visualisation reveals disparities between the spatiotemporal distributions of some dinosaur groups (e.g. sauropodomorphs) and cycads that are inconsistent with co-evolutionary hypotheses. The available data provide no unequivocal support for any of the proposed co-evolutionary interactions between cycads and herbivorous dinosaurs - diffuse co-evolutionary scenarios that are proposed to operate over geological timescales are plausible, but such hypotheses need to be firmly grounded on direct evidence of interaction and may be difficult to support given the patchiness of the fossil record. PMID- 19133961 TI - TRPM2 variants and bipolar disorder risk: confirmation in a family-based association study. AB - OBJECTIVE: Recent case-control studies implicate the transient receptor potential melastatin 2 (TRPM2) channel in conferring risk for bipolar disorder (BD), though the risk variants differed. As confounding effects of population structure could not be unequivocally ruled out as the basis for the discordance, we tested the association of TRPM2 with BD in a family design, which is immune to population stratification, for those TRPM2 single nucleotide polymorphisms (SNPs) previously reported as associated with BD. METHODS: The exon 11 SNP (rs1556314) and four informative intronic SNPs (rs1785437, rs1618355, rs933151, and rs749909) were genotyped in 300 BD families by TaqMan allelic discrimination and results were analyzed using chi(2) test, transmission disequilibrium test, and pedigree-based association. SNP rs1556314 was also genotyped in our case-control sample set comprised of 184 BD and 195 healthy Caucasian subjects. RESULTS: The SNP rs1556314 in exon 11 was significantly associated with bipolar disorder type I (BD-I) (p = 0.011, p(permutation) = 0.015) in the case-control dataset and in the family design (p = 0.018, p(permutation) = 0.052, TDTPHASE). Interestingly, the C T-A haplotype of SNPs rs1618355, rs933151, and rs749909 was significantly associated with early age at onset in BD-I families. CONCLUSION: Significant association of TRPM2 genetic variants with BD in case-control and family datasets further supports a role for TRPM2 in the pathogenesis of this disorder. Overtransmission of the G allele of rs1556314 at exon 11 of TRPM2 in BD-I but not bipolar disorder type II (BD-II) further supports different genetic contributions to the pathogenesis of these bipolar phenotypes. PMID- 19133962 TI - White matter abnormalities in bipolar disorder and schizophrenia detected using diffusion tensor magnetic resonance imaging. AB - OBJECTIVES: Strong qualitative and quantitative evidence exists of white matter abnormalities in both schizophrenia and bipolar disorder (BD). Diffusion tensor imaging (DTI) studies suggest altered connectivity in both disorders. We aim to address the diagnostic specificity of white matter abnormalities in these disorders. METHODS: DTI was used to assess white matter integrity in clinically stable patients with familial BD (n = 42) and familial schizophrenia (n = 28), and in controls (n = 38). Differences in fractional anisotropy (FA) were measured using voxel-based morphometry and automated region of interest analysis. RESULTS: Reduced FA was found in the anterior limb of the internal capsule (ALIC), anterior thalamic radiation (ATR), and in the region of the uncinate fasciculus in patients with BD and those with schizophrenia compared with controls. A direct comparison between patient groups found no significant differences in these regions. None of the findings were associated with psychotropic medication. CONCLUSIONS: Reduced integrity of the ALIC, uncinate fasciculus, and ATR regions is common to both schizophrenia and BD. These results imply an overlap in white matter pathology, possibly relating to risk factors common to both disorders. PMID- 19133963 TI - Eyeblink conditioning anomalies in bipolar disorder suggest cerebellar dysfunction. AB - OBJECTIVES: Accumulating research implicates the cerebellum in non-motor psychological processes and psychiatric diseases, including bipolar disorder (BD). Despite recent evidence that cerebellar lesions have been documented to trigger bipolar-like symptoms, few studies have directly examined the functional integrity of the cerebellum in those afflicted with BD. METHODS: Using a single cue delay eyeblink conditioning procedure, the functional integrity of the cerebellum was examined in 28 individuals with BD (9 manic, 8 mixed, and 11 euthymic) and 28 age-matched healthy controls. RESULTS: Analysis of the bipolar group as a whole indicated a conditioned response acquisition and timing deficit compared to controls. However, when the bipolar group was categorized according to mood state (mixed, manic, euthymic), individuals tested during mixed episodes were strikingly impaired, performing significantly worse than all other groups on both the acquisition and timing of conditioned responses. CONCLUSIONS: These findings extend prior research implicating cerebellar functional abnormalities in BD and suggest that cerebellar dysfunction may be associated with mood state and course of illness. PMID- 19133964 TI - Conceptualizing impulsivity and risk taking in bipolar disorder: importance of history of alcohol abuse. AB - BACKGROUND: Elevated levels of impulsivity and increased risk taking are thought to be core features of both bipolar disorder (BD) and addictive disorders. Given the high rates of comorbid alcohol abuse in BD, alcohol addiction may exacerbate impulsive behavior and risk-taking propensity in BD. Here we examine multiple dimensions of impulsivity and risk taking, using cognitive tasks and self-report measures, in BD patients with and without a history of alcohol abuse. METHODS: Thirty-one BD subjects with a prior history of alcohol abuse or dependence (BD A), 24 BD subjects with no history of alcohol abuse/dependence (BD-N), and 25 healthy control subjects (HC) were assessed with the Barratt Impulsiveness Scale (BIS) and the computerized Balloon Analogue Risk Task (BART). RESULTS: Both BD groups scored significantly higher than controls on the BIS. In contrast, only the BD-A group showed impaired performance on the BART. BD-A subjects popped significantly more balloons than the BD-N and HC groups. In addition, subjects in the BD-A group failed to adjust their performance after popping balloons. Severity of mood symptomatology was not associated with performance on either task. DISCUSSION: The current study supports a primary role of prior alcohol abuse in risk-taking propensity among patients with bipolar disorder. In addition, findings suggest that impulsivity and risky behavior, as operationalized by self-report and experimental cognitive probes, respectively, are separable constructs that tap distinct aspects of the bipolar phenotype. PMID- 19133965 TI - Characterizing impulsivity in mania. AB - OBJECTIVE: To determine whether specific aspects of impulsivity (response disinhibition, inability to delay gratification, inattention) differ between healthy and bipolar manic subjects, and whether these aspects of impulsivity were associated with each other and severity of affective symptoms. METHODS: Performance of 70 bipolar I manic or mixed patients was compared to that of 34 healthy subjects on three tasks specifically designed to study response inhibition, ability to delay gratification, and attention; namely, a stop signal task, a delayed reward task, and a continuous performance task, respectively. Correlations among tasks and with symptom ratings were also performed. RESULTS: Bipolar subjects demonstrated significant deficits on all three tasks as compared to healthy subjects. Performance on the three tasks was largely independent. Task performance was not significantly associated with the severity of affective symptom ratings. However, measures of response inhibition and attention were sensitive to medication effects. Differences in the delayed reward task were independent of medication effects or symptom ratings. During the delayed reward task, although bipolar patients made their choices more slowly than healthy subjects, they were significantly more likely to choose a smaller, but more quickly obtained reward. Moreover, performance on this task was not associated with performance on the other impulsivity measures. Manic patients showed more impulsive responding than mixed patients. CONCLUSIONS: Bipolar I manic patients demonstrate deficits on tests of various aspects of impulsivity as compared to healthy subjects. Some of these differences between groups may be mediated by medication effects. Findings suggested that inability to delay gratification (i.e., delayed reward task) was not simply a result of the speed of decision making or inattention, but rather that it reflected differences between bipolar and healthy subjects in the valuation of reward relative to delay. PMID- 19133966 TI - Comparison of manic and depressive symptoms between children and adolescents with bipolar spectrum disorders. AB - OBJECTIVE: To compare the most severe lifetime (current or past) mood symptoms, duration of illness, and rates of lifetime comorbid disorders among youth with bipolar spectrum disorders [BP (bipolar-I, bipolar-II and bipolar-not otherwise specified)]. METHODS: A total of 173 children (<12 years) with BP, 101 adolescents with childhood-onset BP, and 90 adolescents with adolescent-onset BP were evaluated with standardized instruments. RESULTS: Depression was the most common initial and frequent episode for both adolescent groups, followed by mania/hypomania. Adolescents with childhood-onset BP had the longest illness, followed by children and then adolescents with adolescent-onset BP. Adjusting for sex, socioeconomic status, and duration of illness, while manic, both adolescent groups showed more 'typical' and severe manic symptoms. Mood lability was more frequent in childhood-onset and adolescents with early-onset BP. While depressed, both adolescent groups showed more severe depressive symptoms, higher rates of melancholic and atypical symptoms, and suicide attempts than children. Depressed children had more severe irritability than depressed adolescents. Early BP onset was associated with attention-deficit hyperactivity disorder, whereas later BP onset was associated with panic, conduct, and substance use disorders. Above noted results were similar when each BP subtype was analyzed separately. CONCLUSIONS: Older age was associated with more severe and typical mood symptomatology. However, there were differences and similarities in type, intensity, and frequency of BP symptoms and comorbid disorders related to age of onset and duration of BP and level of psychosocial development. These factors and the normal difficulties youth have expressing and modulating their emotions may explain existing complexities in diagnosing and treating BP in youth, particular in young children, and suggest the need for developmentally sensitive treatments. PMID- 19133967 TI - Emotional hyper-reactivity in normothymic bipolar patients. AB - BACKGROUND: Normothymic states in bipolar disorders are generally considered to be devoid of severe symptoms. However, bipolar patients present subsyndromal symptoms for half of their lives, and often have comorbid psychiatric disorders. If we go beyond the concept of temperamental features, can we identify certain emotional characteristics distinguishing normothymic bipolar patients from normal controls? We previously showed, using self-completed questionnaires, that normothymic bipolar patients display higher levels of emotional lability and intensity than controls. OBJECTIVES: The aim of this study was to assess the emotional reactivity of normothymic bipolar patients, comparing such patients with a normal control group during an experimental mood induction procedure. METHOD: We evaluated the subjective emotional reactivity of 145 subjects (90 control subjects and 55 normothymic bipolar patients), using an emotional induction method based on the viewing of a set of 18 pictures (6 positive, 6 negative, 6 neutral) extracted from the International Affective Picture System. Subjective valence and arousal were recorded with the Self-Assessment Manikin. We also recorded startle reflexes, triggered by a tone occurring during the viewing of two-thirds of the pictures. We controlled for confounding factors, such as concurrent treatments, in all analyses. RESULTS: Normothymic bipolar patients and normal controls assessed valence and arousal similarly for positive and negative images. However, neutral images were considered more pleasant [F(1,143) = 8.4; p = 0.004] and induced a higher level of arousal [F(1,143) = 12.3; p = 0.001] in normothymic bipolar patients than in control subjects. Neutral pictures also triggered a stronger startle reflex in normothymic bipolar patients compared to controls [F(3,123) = 3.1; p = 0.03]. CONCLUSION: Normothymic bipolar patients displayed emotional hyper-reactivity, mostly evidenced in neutral situations. This feature may be linked to emotional dysregulation and is a potential endophenotype and/or a risk factor for bipolar disorders. This trait may be responsible for vulnerability to minor stressful events in everyday life. These findings have potential implications for the daily management of bipolar disorder between crises. PMID- 19133968 TI - A preliminary study of increased amygdala activation to positive affective stimuli in mania. AB - OBJECTIVES: The present study in hypomanic and manic patients explored how amygdala responses to affective stimuli depend on the valence of the stimuli presented. METHODS: We compared 10 patients with 10 matched healthy control subjects. We measured blood oxygen level-dependent (BOLD) responses in the amygdala while subjects passively viewed photographs taken from the International Affective Picture System. After the fMRI session, subjects saw the pictures again and subjectively rated the emotional valence and intensity of each picture. RESULTS: Compared to healthy individuals, hypomanic or manic patients showed higher valence ratings in positive pictures and associated larger BOLD responses in the left amygdala during positive versus neutral picture viewing. This enhanced amygdala activation was correlated with Young Mania Rating Scale scores and with euphoric as opposed to irritable symptom presentation. CONCLUSIONS: Increased valence ratings and amygdala responses to positive affective stimuli may reflect a positive processing bias contributing to elevated mood states characteristic for euphoric mania. PMID- 19133969 TI - Augmentative repetitive navigated transcranial magnetic stimulation (rTMS) in drug-resistant bipolar depression. AB - OBJECTIVES: The efficacy of transcranial magnetic stimulation (TMS) has been poorly investigated in bipolar depression. The present study aimed to assess the efficacy of low-frequency repetitive TMS (rTMS) of the right dorsolateral prefrontal cortex (DLPFC) combined with brain navigation in a sample of bipolar depressed subjects. METHODS: Eleven subjects with bipolar I or bipolar II disorder and major depressive episode who did not respond to previous pharmacological treatment were treated with three weeks of open-label rTMS at 1 Hz, 110% of motor threshold, 300 stimuli/day. RESULTS: All subjects completed the trial showing a statistically significant improvement on the 21-item Hamilton Depression Rating Scale (HAM-D), Montgomery-Asberg Depression Rating Scale, and Clinical Global Impression severity of illness scale (ANOVAs with repeated measures: F = 22.36, p < 0.0001; F = 12.66, p < 0.0001; and F = 10.41, p < 0.0001, respectively). In addition, stimulation response, defined as an endpoint HAM-D score reduction of > or =50% compared to baseline, was achieved by 6 out of 11 subjects, 4 of whom were considered remitters (HAM-D endpoint score < or = 8). Partial response (endpoint HAM-D score reduction between 25% and 50%) was achieved by 3/11 patients. No manic/hypomanic activation was detected during the treatment according to Young Mania Rating Scale scores (ANOVAs with repeated measures: F = 0.62, p = 0.61). Side effects were slight and were limited to the first days of treatment. CONCLUSIONS: Augmentative low-frequency rTMS of the right DLPFC combined with brain navigation was effective and well tolerated in a small sample of drug-resistant bipolar depressive patients, even though the lack of a sham controlled group limits confidence in the results. PMID- 19133970 TI - Polarity at illness onset in bipolar I disorder and clinical course of illness. AB - OBJECTIVES: Studies have suggested that episode polarity at illness onset in bipolar disorder may be predictive of some aspects of lifetime clinical characteristics. We here examine this possibility in a large, well-characterized sample of patients with bipolar I disorder. METHODS: We assessed polarity at onset in patients with bipolar I disorder (N = 553) recruited as part of our ongoing studies of affective disorders. Lifetime clinical characteristics of illness were compared in patients who had a depressive episode at first illness onset (n = 343) and patients who had a manic episode at first illness onset (n = 210). RESULTS: Several lifetime clinical features differed between patients according to the polarity of their onset episode of illness. A logistic regression analysis showed that the lifetime clinical features significantly associated with a depressive episode at illness onset in our sample were: an earlier age at illness onset; a predominantly depressive polarity during the lifetime; more frequent and more severe depressive episodes; and less prominent lifetime psychotic features. CONCLUSIONS: Knowledge of pole of onset may help the clinician in providing prognostic information and management advice to an individual with bipolar disorder. PMID- 19133971 TI - Psychotherapy as monotherapy for the treatment of bipolar II depression: a proof of concept study. AB - OBJECTIVES: We conducted a proof of concept study to determine the feasibility of using an individual psychotherapy, Interpersonal and Social Rhythm Therapy (IPSRT), as monotherapy for the acute treatment of bipolar II depression. METHODS: Unmedicated individuals (n = 17) meeting DSM-IV criteria for bipolar II disorder and currently depressed received weekly psychotherapy (IPSRT) for 12 weeks. After 12 weeks of acute treatment, individuals received an additional 8 weeks of follow-up treatment consisting of continued weekly IPSRT with supplementary lamotrogine for IPSRT non-responders. RESULTS: By week 12, 41% (n = 7) of the sample responded to IPSRT monotherapy (defined as > or =50% reduction in depression scores without an increase in mania scores), 41% (n = 7) dropped out of or were removed from the study, and 18% (n = 3) did not respond to treatment. By week 20, 53% (n = 9) had achieved a response and 29% (n = 5) achieved a full remission of symptoms. CONCLUSIONS: Interpersonal and Social Rhythm Therapy appears to be a promising intervention for a subset of individuals with bipolar II depression. A randomized controlled trial is needed to systematically evaluate the efficacy of IPSRT as an acute monotherapy for bipolar II depression. PMID- 19133973 TI - Resting energy expenditure in manic episode. AB - OBJECTIVE: We aimed to assess the resting energy expenditure in bipolar I disorder, manic episode patients. METHOD: Forty-two bipolar I disorder, manic episode patients that were treated in the inpatient psychiatry clinic of Trakya University Hospital and had met the necessary study criteria were included along with 27 controls. DSM-IV criteria and the Bech-Rafaelsen Mania Rating Scale were used to evaluate patients' diagnosis and severity of the manic episodes. The indirect calorimetry device was used to measure resting energy expenditure values. RESULTS: Resting energy expenditure values of manic patients were found to be higher than those of the controls. Controls showed significant correlations between body mass index and resting energy expenditure, but manic patients did not exhibit similar correlations. There was also no relation between Bech Rafaelsen Mania Rating Scale scores and resting energy expenditure values in manic patients. CONCLUSIONS: We found significantly increased resting energy expenditure values in bipolar I disorder, manic episode patients. These findings suggest a possible clinical use of resting energy expenditure for evaluation of bipolar I disorder manic episode and also suggest resting energy expenditure as a possible biological marker. PMID- 19133972 TI - Functional polymorphism of the glucocorticoid receptor gene associates with mania and hypomania in bipolar disorder. AB - OBJECTIVES: In affective disorders, dysregulation of the hypothalamus-pituitary adrenal (HPA) axis is a frequently observed phenomenon. Subtle changes in glucocorticoid receptor (GR) functioning caused by polymorphisms of the GR gene (NR3C1) may be at the base of the altered reaction of the HPA axis to stress and subsequently related to the development and course of affective disorders. The aim of our study is to evaluate associations between GR gene polymorphisms and bipolar disorder (BD). METHODS: In this study, 245 patients with BD were interviewed to confirm diagnosis and BD subtype. Data on medication use and sociodemographic details were also collected. The control group consisted of 532 healthy blood donors, from which data on sex and age were collected. To perform genotyping, blood was collected from all patients and healthy controls. RESULTS: A trend was found for a protective effect of the exon 9beta polymorphism (p = 0.14) and the TthIIII polymorphism (p < 0.05) on the manifestation of the disease. These effects were significantly influenced by male gender for both polymorphisms. Patients with BD and the A/G variant in exon 9beta had significantly fewer manic and hypomanic episodes than noncarriers (p < 0.05). No further associations were found with the other investigated GR gene polymorphisms and BD. These findings were not corrected for multiple comparisons. CONCLUSIONS: We conclude that the exon 9beta polymorphism and the TthIIII polymorphism of the GR gene may be associated with a protective effect on the clinical manifestation and course in patients with BD. Furthermore, no associations were found between the other studied GR gene polymorphisms and this disease. PMID- 19133974 TI - Letter to the Editor regarding: 'Is bipolar disorder overdiagnosed among patients with substance abuse?'. PMID- 19133977 TI - Histopathological and molecular features of persistent polyclonal B-cell lymphocytosis (PPBL) with progressive splenomegaly. AB - Five cases of persistent polyclonal B-cell lymphocytosis (PPBL) with progressive splenomegaly are reported; three were splenectomized. BCL2/IGH rearrangements were found in three cases; HLA-DRB1*07 in all. Bone marrow (BM) trephines showed a moderate lymphoid infiltrate with intrasinusoidal distribution resembling a splenic marginal-zone lymphoma. Splenic white pulp revealed an enlargement of the marginal-zone area; red pulp was infiltrated by the same lymphocytes engulfing the sinuses. Splenic and BM B-lymphocytes were CD79a(+)/CD20(+)/IgM(+)/IgD(+)/bcl 2(+)/CD27(+)/DBA.44(-)/CD31(-) and polyclonal by immunophenotype/polymerase chain reaction. PPBL features an expansion of splenic marginal-zone B-lymphocytes, which infiltrate BM sinusoids and circulate in the blood with no evidence of clonality, even in cases with progressive splenomegaly. PMID- 19133978 TI - CD4(+)CD25(+)FoxP3(+) regulatory T cells are increased whilst CD3(+)CD4(-)CD8( )alphabetaTCR(+) Double Negative T cells are decreased in the peripheral blood of patients with multiple myeloma which correlates with disease burden. AB - Increased levels of naturally occurring regulatory T cells (T(Reg) cells) have been found in a variety of solid tumours and haematological malignancies. In multiple myeloma (MM), evidence suggests that T(Reg) cells are increased though controversy exists with regards to their function and no relationship to disease stage and treatment has been demonstrated. Here, we demonstrate significantly elevated levels of functional CD4(+)CD25(+)FoxP3(+) T(Reg) cells in a large cohort of patients with MM as well as monoclonal gammopathy of uncertain significance (MGUS) in comparison to age-matched, healthy controls. The frequency of Double Negative T(Reg) cells was also evaluated, demonstrating that these cells were reduced in patients with MM. Furthermore, a characteristic profile of immunomodulatory cytokines in the peripheral blood and bone marrow of patients with MM and MGUS was demonstrated, compared with healthy controls. This data adds further evidence to the understanding of the role of T(Reg) cell subsets in tumour immunology and the fundamentals of the host/tumour immune conflict. PMID- 19133979 TI - The protein C omega-loop substitution Asn2Ile is associated with reduced protein C anticoagulant activity. AB - We report a kindred with heritable protein C (PC) deficiency in which two siblings with severe thrombosis showed a composite type I and IIb PC deficiency phenotype, identified using commercial PC assays (proband: PC antigen 42 u/dl, amidolytic activity 40 u/dl, anticoagulant activity 9 u/dl). The independent PROC nucleotide variations c.669C>A (predictive of Ser181Arg) and c.131C>T (predictive of Asn2Ile) segregated with the type I and type IIb PC deficiency phenotypes respectively, but co-segregated in the siblings with severe thrombosis. Soluble thrombomodulin (sTM)-mediated inhibition of plasma thrombin generation from an individual with PC-Asn2Ile was lower (endogenous thrombin potential (ETP) 56 +/- 1% that of ETP determined without sTM) than control plasma (ETP 15 +/- 2%) indicating reduced PC anticoagulant activity. Recombinant APC-Asn2Ile exhibited normal amidolytic activity but impaired anticoagulant activity. Protein S (PS) dependent anticoagulant activity of recombinant APC-Asn2Ile and binding of recombinant APC-Asn2Ile to endothelial protein C receptor (EPCR) were reduced compared to recombinant wild-type APC. Asn2 lies within the omega-loop of the PC/APC Gla domain and this region is critical for calcium-induced folding and subsequent interactions with anionic phospholipids, EPCR and PS. The disruption of these interactions in this naturally-occurring PC variant highlights their collective importance in mediating APC anticoagulant activity in vivo. PMID- 19133981 TI - Decitabine is an effective treatment of idiopathic myelofibrosis. PMID- 19133980 TI - Glucocorticoid resistance in a multiple myeloma cell line is regulated by a transcription elongation block in the glucocorticoid receptor gene (NR3C1). AB - Glucocorticoid (GC) effects are mediated by the glucocorticoid receptor (GR). Several studies have demonstrated that a lower number of receptors per cell were associated with poor GC response. The regulation of GR expression is complex; the levels of GR can be autologously regulated by its ligand and also by transcriptional, post-transcriptional and post-translational mechanisms. Using three human myeloma cell lines that parallel the development of GC resistance, this work describes the mechanism involved in the downregulation of GR expression. The decreased expression was neither due to mutations in the gene encoding GR, NR3C1, nor due to methylation of the promoters. A gradual decrease in NR3C1 transcripts was seen during the development of resistance, the level of expression of exon 1 to 2 RNA fragments remained the same in sensitive and resistant cell lines but a chromatin immunoprecipitation assay demonstrated that RNA polymerase II, detectable throughout exon 2 to 3 in the sensitive cells, was undetectable on exon 3 in the resistant variant, suggesting lower or no transcription at this site. These studies demonstrated that downregulation of NR3C1 mRNA in a resistant cell line involves a block to transcriptional elongation within intron B of NR3C1. This block may represent an important element in the regulation of GR expression. PMID- 19133982 TI - Role of TLX1 in T-cell acute lymphoblastic leukaemia pathogenesis. PMID- 19133983 TI - Fluorescence-activated cell sorting (FACS) of whole mount in situ hybridization (WISH) labelled haematopoietic cell populations in the zebrafish. AB - The zebrafish is a robust animal model for studying vertebrate haematopoiesis and immune cell interactions. However, fluorescence activated cell sorting (FACS) has been limited due to a paucity of available functional zebrafish antibodies. We have developed a technique combining FACS with whole mount in situ hybridization (WISH) that enables the sorting and examining of fixed zebrafish blood cell populations at different stages of embryonic development, providing the opportunity to correlate RNA expression data with cellular morphology. PMID- 19133984 TI - Persistent (current) in the face of adversity ... a new class of cardiac anti ischaemic compounds on the horizon? AB - Although a persistent component of the sodium current (INaP) was described in cardiac tissue about three decades ago, its physiological role and potential as a therapeutic target was not immediately apparent. Subsequent demonstrations that INaP is enhanced by hypoxia and ischaemia, and that Na+ influx via INaP may contribute to cellular damage, diastolic dysfunction and arrhythmias during ischaemia and reperfusion, raised interest in INaP as a target for anti-ischaemic drugs. Several agents have now been developed to clinical stages, which have INaP block as either their main action, or as a useful co-effect. In this issue of the British Journal of Pharmacology, Vacher et al. report the anti-ischaemic actions of F15845, which appears to exhibit the most selective block of INaP yet described. Its efficacy in animal models of angina raises the prospect of new, specific, INaP blockers that may represent a largely unexploited opportunity for a new class of anti-ischaemic compounds. PMID- 19133985 TI - F 15845 inhibits persistent sodium current in the heart and prevents angina in animal models. AB - BACKGROUND AND PURPOSE: Activation of the persistent sodium current in ischaemic myocardium results in calcium overload which is toxic for the cardiomyocyte. Thus, the activity of 3-(R)-[3-(2-methoxyphenylthio-2-(S)-methylpropyl]amino-3,4 dihydro-2H-1,5 benzoxathiepine bromhydrate (F 15845), a new selective persistent sodium current blocker, in protecting against the effects of cardiac ischaemia was examined, in both in vitro and in vivo models. EXPERIMENTAL APPROACH: Electrophysiological studies using patch-clamp and conventional microlelectrode techniques, isolated perfused hearts and models of angina in anaesthetized animals were used to assess the protection afforded by F 15845 against ischaemia induced changes. KEY RESULTS: F 15845 reduced the persistent sodium current activated by veratridine (IC(50) 1.58 x 10(-6) mol.L(-1)). F 15845 blocked voltage-gated human cardiac sodium channels in a novel, voltage-dependent manner, selectively affecting steady-state inactivation. F 15845 did not affect action potential shape and basal function of guinea pig isolated perfused hearts but did reduce ischaemia-induced diastolic contracture in this model (IC(50) 0.64 x 10( 6) mol.L(-1)). In rabbits, F 15845 given i.v. (ED(50) 0.05 mg.kg(-1)) or orally (ED(50) 0.13 mg.kg(-1)) dose-dependently and powerfully inhibited regional myocardial ischaemia-induced ST segment elevation in the absence of haemodynamic effects, implying direct cardiac activity. In dogs, F 15845 dose-dependently inhibited epicardial ST segment changes (70 +/- 8% at 0.63 mg.kg(-1)) in an experimental angina model of demand ischaemia, again without haemodynamic effects, confirming a direct anti-anginal activity. CONCLUSIONS AND IMPLICATIONS: F 15845 is a selective, potent blocker of the persistent sodium current, generated by the human Na(v)1.5 channel isoforms, and prevents cardiac angina in animal models. PMID- 19133986 TI - Continuity and change. PMID- 19133987 TI - Allopurinol for pain relief: more than just crystal clearance? AB - Gout and pain are synonymous, and a study in this issue of the BJP reports a novel anti-nociceptive effect of allopurinol, the drug most commonly used to treat gout. Allopurinol works by inhibiting xanthine oxidase (XO), the enzyme responsible for converting hypoxanthine to uric acid which is deposited as crystals in the joints of gout sufferers. Hypoxanthine is a metabolite of, and a possible precursor to, adenosine. Schmidt et al., find that acute inhibition of XO with allopurinol produces a modest adenosine A(1) receptor-mediated anti nociceptive effect in common tests of chemical and thermal nociception in mice. A concomitant increase in cerebrospinal fluid levels of adenosine supports their hypothesis that inhibiting XO increases adenosine levels via salvage from hypoxanthine. Elevating endogenous adenosine levels by inhibiting metabolism is a well-established strategy for producing anti-nociception in many preclinical models, but inhibiting XO is likely to be particularly beneficial in some chronic pain states because of the pro-nociceptive reactive oxygen species that are produced by XO activity. Thus, allopurinol may have unexpected benefits in pain associated with chronic inflammation, diabetes and vascular dysfunction. PMID- 19133989 TI - Modelling the interdependence between the stoichiometry of receptor oligomerization and ligand binding for a coexisting dimer/tetramer receptor system. AB - Many G protein-coupled receptors have been shown to exist as oligomers, but the oligomerization state and the effects of this on receptor function are unclear. For some G protein-coupled receptors, in ligand binding assays, different radioligands provide different maximal binding capacities. Here we have developed mathematical models for co-expressed dimeric and tetrameric species of receptors. We have considered models where the dimers and tetramers are in equilibrium and where they do not interconvert and we have also considered the potential influence of the ligands on the degree of oligomerization. By analogy with agonist efficacy, we have considered ligands that promote, inhibit or have no effect on oligomerization. Cell surface receptor expression and the intrinsic capacity of receptors to oligomerize are quantitative parameters of the equations. The models can account for differences in the maximal binding capacities of radioligands in different preparations of receptors and provide a conceptual framework for simulation and data fitting in complex oligomeric receptor situations. PMID- 19133990 TI - Targeting V1A-vasopressin receptors with [Arg6, D-Trp7,9, NmePhe8]-substance P (6 11) identifies a strategy to develop novel anti-cancer therapies. AB - BACKGROUND AND PURPOSE: The anti-cancer agent [Arg(6), D-Trp(7,9), N(me)Phe(8)] substance P (6-11) (SP-G) modulates gastrin releasing peptide (GRP) and arginine vasopressin signalling in small cell lung cancer cells leading to growth arrest and apoptosis. We have shown that SP-G acts as a biased agonist at GRP receptors. This work examines the hypothesis that SP-G acts as a biased agonist at the V(1A) vasopressin receptor. EXPERIMENTAL APPROACH: The human V(1A) receptor was expressed in CHO-K1 cells. Extracellular regulated kinase (ERK) activation and intracellular Ca(2+) were measured using activation state-specific antibodies and Fura-2-AM respectively. The effect of SP-G on tumourigenicity was assessed by colony assay. KEY RESULTS: In V(1A) receptor expressing cells, SP-G caused a sustained activation of ERK via a stimulation of V(1A) receptor coupling to G(i). Inhibition of G(i) with Pertussis toxin attenuated the inhibition by SP-G of the growth of CHO-K1 cells stably expressing the V(1A) receptor. Chimeric V(1A) receptors containing the second or third intracellular loop of the V(2) receptor were capable of binding vasopressin and SP-G but had altered ability to activate phospholipase C (PLC) and ERK. The second intracellular loop of the V(1A) receptor was essential for vasopressin-stimulated PLC and ERK activation but not for SP-G-induced ERK activation. CONCLUSIONS AND IMPLICATIONS: This work provides mechanistic insight, for biased agonists at V(1A) receptors and highlights a potential role for such agents as anti-cancer agents. PMID- 19133991 TI - A standardized extract of Ginkgo biloba suppresses doxorubicin-induced oxidative stress and p53-mediated mitochondrial apoptosis in rat testes. AB - BACKGROUND AND PURPOSE: Doxorubicin evokes oxidative stress and precipitates cell apoptosis in testicular tissues. The aim of this study was to investigate whether the Ginkgo biloba extract 761 (EGb), a widely used herbal medicine with potent anti-oxidant and anti-apoptotic properties, could protect testes from such doxorubicin injury. EXPERIMENTAL APPROACH: Sprague-Dawley male rats (8 weeks old) were given vehicle, doxorubicin alone (3 mg kg(-1) every 2 days for three doses), EGb alone (5 mg kg(-1) every 2 days for three doses), or EGb followed by doxorubicin (each dose administered 1 day after EGb). At 7 days after the first drug treatment oxidative and apoptotic testicular toxicity was evaluated by biochemical, histological and flow cytometric analyses. KEY RESULTS: Compared with controls, testes from doxorubicin-treated rats displayed impaired spermatogenesis, depleted haploid germ cell subpopulations, increased lipid peroxidation products (malondialdehyde), depressed antioxidant enzyme activities (superoxide dismutase, glutathione peroxidase and glutathione), reduced antioxidant enzyme expression (superoxide dismutase) and elevated apoptotic indexes (pro-apoptotic modulation of Bcl-2 family proteins, intensification of p53 and Apaf-1, release of mitochondrial cytochrome c, activation of caspase-3 and increase of terminal deoxynucleotidyl transferase nick-end labelling/sub haploid cells), while EGb pretreatment effectively alleviated all of these doxorubicin-induced abnormalities in testes. CONCLUSIONS AND IMPLICATIONS: These results demonstrate that EGb protected against the oxidative and apoptotic actions of doxorubicin on testes. EGb may be a promising adjuvant therapy medicine, potentially ameliorating testicular toxicity of this anti-neoplastic agent in clinical practice. PMID- 19133988 TI - Nuclear receptors as therapeutic targets in cholestatic liver diseases. AB - Cholestasis results in intrahepatic accumulation of cytotoxic bile acids, which cause liver damage ultimately leading to biliary fibrosis and cirrhosis. Cholestatic liver injury is counteracted by a variety of adaptive hepatoprotective mechanisms including alterations in bile acid transport, synthesis and detoxification. The underlying molecular mechanisms are mediated mainly at a transcriptional level via a complex network involving nuclear receptors including the farnesoid X receptor, pregnane X receptor, vitamin D receptor and constitutive androstane receptor, which target overlapping, although not identical, sets of genes. Because the intrinsic adaptive response to bile acids cannot fully prevent liver injury in cholestasis, therapeutic targeting of these receptors via specific and potent agonists may further enhance the hepatic defence against toxic bile acids. Activation of these receptors results in repression of bile acid synthesis, induction of phases I and II bile acid hydroxylation and conjugation and stimulation of alternative bile acid export while limiting hepatocellular bile acid import. Furthermore, the use of nuclear receptor ligands may not only influence bile acid transport and metabolism but may also directly target hepatic fibrogenesis and inflammation. Many drugs already used to treat cholestasis and its complications such as pruritus (e.g. ursodeoxycholic acid, rifampicin, fibrates) may act via activation of nuclear receptors. More specific and potent nuclear receptor ligands are currently being developed. This article will review the current knowledge on nuclear receptors and their potential role in the treatment of cholestatic liver diseases. PMID- 19133992 TI - Inotropy and L-type Ca2+ current, activated by beta1- and beta2-adrenoceptors, are differently controlled by phosphodiesterases 3 and 4 in rat heart. AB - BACKGROUND AND PURPOSE: beta(1)- and beta(2)-adrenoceptors coexist in rat heart but beta(2)-adrenoceptor-mediated inotropic effects are hardly detectable, possibly due to phosphodiesterase (PDE) activity. We investigated the influence of the PDE3 inhibitor cilostamide (300 nmol x L(-1)) and the PDE4 inhibitor rolipram (1 micromol x L(-1)) on the effects of (-)-catecholamines. EXPERIMENTAL APPROACH: Cardiostimulation evoked by (-)-noradrenaline (ICI118551 present) and ( )-adrenaline (CGP20712A present) through beta(1)- and beta(2)-adrenoceptors, respectively, was compared on sinoatrial beating rate, left atrial and ventricular contractile force in isolated tissues from Wistar rats. L-type Ca(2+) current (I(Ca-L)) was assessed with whole-cell patch clamp. KEY RESULTS: Rolipram caused sinoatrial tachycardia. Cilostamide and rolipram did not enhance chronotropic potencies of (-)-noradrenaline and (-)-adrenaline. Rolipram but not cilostamide potentiated atrial and ventricular inotropic effects of (-) noradrenaline. Cilostamide potentiated the ventricular effects of (-)-adrenaline but not of (-)-noradrenaline. Concurrent cilostamide + rolipram uncovered left atrial effects of (-)-adrenaline. Both rolipram and cilostamide augmented the (-) noradrenaline (1 micromol x L(-1)) evoked increase in I(Ca-L). (-)-Adrenaline (10 micromol x L(-1)) increased I(Ca-L) only in the presence of cilostamide but not rolipram. CONCLUSIONS AND IMPLICATIONS: PDE4 blunts the beta(1)-adrenoceptor mediated inotropic effects. PDE4 reduces basal sinoatrial rate in a compartment distinct from compartments controlled by beta(1)- and beta(2)-adrenoceptors. PDE3 and PDE4 jointly prevent left atrial beta(2)-adrenoceptor-mediated inotropy. Both PDE3 and PDE4 reduce I(Ca-L) responses through beta(1)-adrenoceptors but the PDE3 component is unrelated to inotropy. PDE3 blunts both ventricular inotropic and I(Ca-L) responses through beta(2)-adrenoceptors. PMID- 19133993 TI - The vascular effects of different arginase inhibitors in rat isolated aorta and mesenteric arteries. AB - BACKGROUND AND PURPOSE: Arginase and nitric oxide (NO) synthase share the common substrate L-arginine, and arginase inhibition is proposed to increase NO production by increasing intracellular levels of L-arginine. Many different inhibitors are used, and here we have examined the effects of these inhibitors on vascular tissue. EXPERIMENTAL APPROACH: Each arginase inhibitor was assessed by its effects on isolated rings of aorta and mesenteric arteries from rats by: (i) their ability to preserve the tolerance to repeated applications of the endothelium-dependent agonist acetylcholine (ACh); and (ii) their direct vasorelaxant effect. KEY RESULTS: In both vessel types, tolerance (defined as a reduced response upon second application) to ACh was reversed with addition of L arginine, (S)-(2-boronethyl)-L-cysteine HCl (BEC) or N(G)-Hydroxy-L-arginine (L NOHA). On the other hand, N(omega)-hydroxy-nor-L-arginine (nor-NOHA) significantly augmented the response to ACh, an effect that was partially reversed with L-arginine. No effect on tolerance to ACh was observed with L valine, nor-valine or D,L, alpha-difluoromethylornithine (DFMO). BEC, L-NOHA and nor-NOHA elicited endothelium-independent vasorelaxation in both endothelium intact and denuded aorta while L-valine, DFMO and nor-valine did not. CONCLUSIONS AND IMPLICATIONS: BEC and L-NOHA, but not nor-NOHA, L-valine, DFMO or nor-valine, significantly reversed tolerance to ACh possibly conserving L-arginine levels and therefore increasing NO bioavailability. However, both BEC and L-NOHA caused endothelium-independent vasorelaxation in rat aorta, suggesting that these inhibitors have a role beyond arginase inhibition alone. Our data thus questions the interpretation of many studies using these antagonists as specific arginase inhibitors in the vasculature, without verification with other methods. PMID- 19133995 TI - VIP modulates the pro-inflammatory maternal response, inducing tolerance to trophoblast cells. AB - BACKGROUND AND PURPOSE: Successful embryo implantation is followed by a local pro inflammatory and Th1 response, subsequently controlled by a Th2 response. Vasoactive intestinal peptide (VIP) has anti-inflammatory effects and promotes tolerogenic/Th2 responses while favouring embryonic development. We investigated the potential regulatory role of VIP on human trophoblast cells, maternal pro inflammatory responses and trophoblast-maternal leukocyte interactions. EXPERIMENTAL APPROACH: We tested VIP effects directly on a trophoblast cell line (Swan 71 cells) and after co-culture with maternal peripheral blood mononuclear cells (PBMCs) as models of the feto-maternal dialogue. We also co-cultured maternal and paternal PBMCs to test effects of endogenous VIP on maternal alloresponses. KEY RESULTS: Swan 71 cells express VPAC(1) receptors and VIP induced their proliferation and the expression of leukaemia inhibitor factor, a pro-implantatory marker. After interaction with trophoblast cells, VIP increased Foxp3, the proportion of CD4+CD25+Foxp3+ cells within maternal PBMCs and transforming growth factor beta expression. Also, during the trophoblast-maternal PBMCs interaction, VIP reduced pro-inflammatory mediators [interleukin (IL)-6, monocyte chemoattractant protein 1, nitric oxide], while increasing IL-10. Trophoblast cells produced VIP which dose-dependently suppressed allomaternal responses, accompanied by reduced expression of the T cell transcription factor, T-bet. CONCLUSIONS AND IMPLICATIONS: Vasoactive intestinal peptide induced pro implantatory markers and trophoblast cell proliferation, while controlling the initial pro-inflammatory response, by increasing maternal regulatory T cells and anti-inflammatory cytokines. As an autocrine regulatory peptide VIP might contribute to fetal survival through two mechanisms; a direct trophic effect on trophoblast cells and an immunomodulatory effect that favours tolerance to fetal antigens. PMID- 19133994 TI - Acute hypertension reveals depressor and vasodilator effects of cannabinoids in conscious rats. AB - BACKGROUND AND PURPOSE: The cardiovascular effects of cannabinoids can be influenced by anaesthesia and can differ in chronic hypertension, but the extent to which they are influenced by acute hypertension in conscious animals has not been determined. EXPERIMENTAL APPROACH: We examined cardiovascular responses to intravenous administration of anandamide and the synthetic cannabinoid, (R)-(+) [2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6 yl]-1-naphthalenylmethanone (WIN55212-2), in conscious male Wistar rats made acutely hypertensive by infusion of angiotensin II (AII) and arginine vasopressin (AVP). Rats were chronically instrumented for measurement of arterial blood pressure and vascular conductances in the renal, mesenteric and hindquarters beds. KEY RESULTS: Anandamide dose-dependently decreased the mean arterial blood pressure of rats made hypertensive by AII-AVP infusion, but not normotensive rats. Interestingly, acute hypertension also revealed a hypotensive response to WIN55212-2, which caused hypertension in normotensive animals. The enhanced depressor effects of the cannabinoids in acute hypertension were associated with increased vasodilatation in hindquarters, renal and mesenteric vascular beds. Treatment with URB597, which inhibits anandamide degradation by fatty acid amide hydrolase, potentiated the depressor and mesenteric vasodilator responses to anandamide. Furthermore, haemodynamic responses to WIN55212-2, but not to anandamide, were attenuated by the CB(1) receptor antagonist, AM251 [N-(piperidin 1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophen yl)-4-methyl-1H-pyrazole-3 carboxamide]. CONCLUSIONS AND IMPLICATIONS: These results broadly support the literature showing that the cardiovascular effects of cannabinoids can be exaggerated in hypertension, but highlight the involvement of non-CB(1) receptor mediated mechanisms in the actions of anandamide. PMID- 19133996 TI - Do gene polymorphisms alone or in combination affect the function of human beta3 adrenoceptors? AB - BACKGROUND AND PURPOSE: beta(3)-Adrenoceptors mediate many important physiological functions, for example, in the urinary bladder. The corresponding gene is polymorphic, and the W64R (Trp64Arg) single nucleotide polymorphism has been associated with disease states such as obesity, type 2 diabetes and bladder dysfunction. While these clinical data suggest that the 64R variant is hypofunctional, previous in vitro studies in which this variant was generated by site-directed mutagenesis and subsequent transfection have not consistently confirmed this. EXPERIMENTAL APPROACH: We transfected the wild-type human beta(3) adrenoceptor and the 64R variant and also the more recently discovered 265M and 306F variants as well as 64R/265M and 64R/306F double mutants into human embryonic kidney cells and selected clones expressing the receptors at a density of about 100 fmol mg protein(-1). Receptor activation was measured by cAMP accumulation and ligand affinity by radioligand binding. Desensitisation was assessed as alterations of cAMP responses after prolonged agonist treatment. KEY RESULTS: Neither mutated receptor exhibited alterations in efficacy or potency for cAMP accumulation for any of five agonists (isoprenaline, noradrenaline, YM 178, FK 4664, CGP 12 177). In competition binding studies, the mutations did not affect the ability of any agonist to bind to the receptor. Wild-type receptors and the 64R variant exhibited similar isoprenaline-induced functional desensitization during a 24 h treatment. CONCLUSIONS AND IMPLICATIONS: None of the polymorphisms tested here significantly altered the interaction of isoprenaline, noradrenaline, YM 178, FK 4664 or CGP 12 177 with the human beta(3) adrenoceptor when expressed at near physiological levels in a human cell line. PMID- 19133997 TI - Anti-nociceptive properties of the xanthine oxidase inhibitor allopurinol in mice: role of A1 adenosine receptors. AB - BACKGROUND AND PURPOSE: Allopurinol is a potent inhibitor of the enzyme xanthine oxidase, used primarily in the treatment of hyperuricemia and gout. It is well known that purines exert multiple effects on pain transmission. We hypothesized that the inhibition of xanthine oxidase by allopurinol, thereby reducing purine degradation, could be a valid strategy to enhance purinergic activity. The aim of this study was to investigate the anti-nociceptive profile of allopurinol on chemical and thermal pain models in mice. EXPERIMENTAL APPROACH: Mice received an intraperitoneal (i.p.) injection of vehicle (Tween 10%) or allopurinol (10-400 mg kg(-1)). Anti-nociceptive effects were measured with intraplantar capsaicin, intraplantar glutamate, tail-flick or hot-plate tests. KEY RESULTS: Allopurinol presented dose-dependent anti-nociceptive effects in all models. The opioid antagonist naloxone did not affect these anti-nociceptive effects. The non selective adenosine-receptor antagonist caffeine and the selective A(1) adenosine receptor antagonist, DPCPX, but not the selective A(2A) adenosine-receptor antagonist, SCH58261, completely prevented allopurinol-induced anti-nociception. No obvious motor deficits were produced by allopurinol, at doses up to 200 mg kg( 1). Allopurinol also caused an increase in cerebrospinal fluid levels of purines, including the nucleosides adenosine and guanosine, and decreased cerebrospinal fluid concentration of uric acid. CONCLUSIONS AND IMPLICATIONS: Allopurinol induced anti-nociception may be related to adenosine accumulation. Allopurinol is an old and extensively used compound and seems to be well tolerated with no obvious central nervous system toxic effects at high doses. This drug may be useful to treat pain syndromes in humans. PMID- 19133998 TI - Galantamine improves apomorphine-induced deficits in prepulse inhibition via muscarinic ACh receptors in mice. AB - BACKGROUND AND PURPOSE: Galantamine, a weak acetylcholine esterase (AChE) inhibitor and allosteric potentiator of nicotinic ACh receptors (nAChRs), improves apomorphine-induced deficits in prepulse inhibition (PPI), sensory information-processing deficits, via a nAChR-independent mechanism. The present study examined the role of muscarinic ACh receptors (mAChRs) in the effect of galantamine, and studied the mechanism of galantamine-induced increases in prefrontal ACh levels in mice. EXPERIMENTAL APPROACH: Apomorphine (1 mg kg(-1)) was administered to male ddY mice (9-10 weeks old) to create a PPI deficit model. Extracellular ACh concentrations in the prefrontal cortex were measured by in vivo microdialysis. KEY RESULTS: Galantamine- and donepezil-mediated improvements in apomorphine-induced PPI deficits were blocked by the preferential M(1) mAChR antagonist telenzepine. The mAChR agonist oxotremorine also improved apomorphine induced PPI deficits. Galantamine, like donepezil, increased extracellular ACh concentrations in the prefrontal cortex. Galantamine-induced increases in prefrontal ACh levels were partially blocked by the dopamine D(1) receptor antagonist SCH23390, but not by antagonists of mAChRs (telenzepine) and nAChRs (mecamylamine). Galantamine increased dopamine, but not 5-HT, release in the prefrontal cortex. CONCLUSIONS AND IMPLICATIONS: Galantamine improves apomorphine induced PPI deficits by stimulating mAChRs through increasing brain ACh levels via a dopamine D(1) receptor-dependent mechanism and AChE inhibition. PMID- 19134000 TI - Inhibition of the phosphatase PTEN protects mice against oleic acid-induced acute lung injury. AB - BACKGROUND AND PURPOSE: Injury to the lung parenchyma is a constitutional feature shared by many lung diseases. The protein, phosphatase and tensin homologue deleted on chromosome Ten (PTEN) is a major suppressor of phosphoinositide-3 kinase/Akt signalling, a vital survival pathway in lung parenchymal cells. Based on this, we hypothesized that PTEN inhibition in vivo would enhance cell tolerance to stress thereby preventing acute lung injury. EXPERIMENTAL APPROACH: We evaluated the ability of a PTEN inhibitor, potassium bisperoxo (1,10 phenanthroline) oxovanadate [bpV(phen)], to prevent acute lung injury induced by oleic acid (OA) in adult C57BL/6 mice. Lung assessments included bronchoalveolar lavage, tissue morphology, immunostaining for markers of cell death, cell identity, phospho-Akt and phospho-ERK levels and oximetry. KEY RESULTS: OA induced acute lung injury in a dose- and time-dependent manner. No injury was observed in the vehicle control or bpV(phen) treatment groups. PTEN inhibition by bpV(phen) increased lung tissue levels of phospho-Akt and ERK and but not focal adhesion kinase. This occurred in conjunction with a statistically significant reduction in protein content, lactate dehydrogenase, as well as tumour necrosis factor-alpha and chemokines in bronchoalveolar lavage fluid when compared with OA treatment alone. The incidence of alveolar lesions, consistent with acute lung injury, and terminal uridine deoxynucleotidyl transferase dUTP nick end labelling (TUNEL)-positive cells was also significantly reduced. Importantly, PTEN suppression maintained pulmonary function. CONCLUSIONS AND IMPLICATIONS: Treatment with bpV(phen) significantly reduced the severity of acute lung injury in mice indicating that additional investigation is warranted to understand the important role that this phosphatase may play in the lung. PMID- 19133999 TI - 5-HT1A receptors are involved in the cannabidiol-induced attenuation of behavioural and cardiovascular responses to acute restraint stress in rats. AB - BACKGROUND AND PURPOSE: Cannabidiol (CBD) is a non-psychotomimetic compound from Cannabis sativa which induces anxiolytic- and antipsychotic-like effects in rodents. These effects could be mediated by facilitation of the endocannabinoid system or by the activation of 5-HT(1A) receptors. As either of these mechanisms could promote adaptation to inescapable stress, the aim of the present work was to test the hypothesis that CBD would attenuate the autonomic and behavioural consequences of restraint stress (RS). We also investigated if the responses to CBD depended on activation of 5-HT(1A) receptors. EXPERIMENTAL APPROACH: Male Wistar rats received i.p. injections of vehicle or CBD (1, 10 or 20 mg kg(-1)) and 30 min later were submitted to 60 min of restraint where their cardiovascular responses were recorded. The protocol of the second experiment was similar to the first one except that animals received i.p. injections of the 5-HT(1A) receptor antagonist WAY100635 (0.1 mg kg(-1)) before CBD treatment and exposure to restraint. 24 h later they were also tested in the elevated plus-maze (EPM), an animal model of anxiety. KEY RESULTS: Exposure to RS increased blood pressure and heart rate and induced an anxiogenic response in the EPM 24 h later. These effects were attenuated by CBD. WAY100635 by itself did not change the cardiovascular and anxiogenic response to RS, but blocked the effects of CBD. CONCLUSION AND IMPLICATIONS: The results suggest that CBD can attenuate acute autonomic responses to stress and its delayed emotional consequences by facilitating 5-HT(1A) receptor-mediated neurotransmission. PMID- 19134002 TI - Analysis of the repressor element-1 silencing transcription factor/neuron restrictive silencer factor occupancy of non-neuronal genes in peripheral lymphocytes from patients with Huntington's disease. AB - We have previously demonstrated that the transcription of neuronal repressor element-1/neuron-restrictive silencer element (RE1/NRSE)-regulated genes is reduced in the brain of subjects with Huntington's disease (HD) as a result of increased binding of the repressor element-1 silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) to its RE1/NRSE targets. As specific non-neuronal REST/NRSF-regulated genes have been identified in the human genome, we exploited the possibility that the binding of REST/NRSF to its target RE1/NRSE sites may also be altered in the peripheral tissues of HD patients. Our results show that REST/NRSF occupancy is increased in lymphocytes from HD subjects, thus indicating for the first time that the activity of the RE1/NRSE sites is dysfunctional in vivo. Chromatin immunoprecipitation (ChIP) of the RE1/NRSE sites in lymphocytes may therefore be a reproducible, sensitive and specific means of searching for candidate markers of HD onset and progression. PMID- 19134001 TI - Etanercept prevents airway hyperresponsiveness by protecting neuronal M2 muscarinic receptors in antigen-challenged guinea pigs. AB - BACKGROUND AND PURPOSE: Increased tumour necrosis factor-alpha (TNF-alpha) is associated with airway hyperreactivity in antigen-challenged animals. In human asthmatics, TNF-alpha is increased and blocking it prevents airway hyperreactivity in some asthmatic patients. However, the mechanisms by which TNF alpha mediates hyperreactivity are unknown. Airway hyperreactivity can be caused by dysfunction of neuronal M(2) muscarinic receptors that normally limit acetylcholine release from parasympathetic nerves. Here we test whether blocking TNF-alpha receptors with etanercept prevents M(2) receptor dysfunction and airway hyperreactivity in antigen-challenged guinea pigs. EXPERIMENTAL APPROACH: Ovalbumin-sensitized guinea pigs were challenged by inhalation of antigen. Some animals received etanercept (3 mg kg(-1) i.p.) 3 h before challenge. 24 h after challenge, airway hyperreactivity and M(2) receptor function were tested. Inflammatory cells in bronchoalveolar lavage, blood and lung were counted. TNF alpha and its receptors were detected by real-time RT-PCR and immunocytochemistry in parasympathetic nerves from humans and guinea pigs and in human neuroblastoma cells. KEY RESULTS: Antigen-challenged animals were hyperreactive to vagal stimulation and neuronal M(2) receptors were dysfunctional. Both M(2) receptor dysfunction and airway hyperreactivity were prevented by etanercept. Etanercept reduced eosinophils around airway nerves, and in blood, bronchoalveolar lavage and airway smooth muscle. Also, TNF-alpha decreased M(2) receptor mRNA in human and guinea pig parasympathetic neurons. CONCLUSIONS AND IMPLICATIONS: Tumour necrosis factor-alpha may contribute to M(2) receptor dysfunction and airway hyperreactivity directly by decreasing receptor expression and indirectly by promoting recruitment of eosinophils, containing major basic protein, an M(2) antagonist. This suggests that etanercept may be beneficial in treatment of allergic asthma. PMID- 19134003 TI - Reduction of cerebral oxidative stress following environmental enrichment in mice with Alzheimer-like pathology. AB - Oxidative stress is a key feature during progression of chronic neurodegenerative conditions such as Alzheimer's disease. In aging humans and animals, voluntary exercise lowers oxidative stress reactions. Additionally, recent work in our lab demonstrated that cognitive and physical stimulation (termed environmental enrichment) counteracts amyloid beta pathology, neurovascular dysfunction and behavioral symptoms in mice with Alzheimer-like disease. Based on these facts, we hypothesized that cognitive and physical activity can also protect against oxidative stress in Alzheimer-diseased brain. We, therefore, kept female TgCRND8 mice under standard and enriched housing from day 30 until 5 months of age. Environmental stimulation attenuated pro-oxidative processes and triggered anti oxidative defense mechanisms as indicated by diminished biomarkers for reactive oxygen and nitrogen species, downregulation of pro-inflammatory and pro-oxidative mediators, decreased expression of pro-apoptotic caspases, and upregulation of SOD1 and SOD2. This study identifies a thus far undescribed antagonizing effect of environmental stimulation on Alzheimer's disease-related oxidative damage. PMID- 19134004 TI - Involvement of molecular mimicry between human T-cell leukemia virus type 1 gp46 and osteoprotegerin in induction of hypercalcemia. AB - Human T-cell leukemia virus type-1 (HTLV-1) causes adult T-cell leukemia/lymphoma (ATL), frequently associated with hypercalcemia and bone destruction. A positive correlation between the appearance of an antibody recognizing the central region (Asp197 to Leu216) on Gp46, gp46-197, and the severity of ATL has been demonstrated. In this study, five male Nihon Hakusyoku rabbits were immunized with a synthetic peptide corresponding to the gp46-197 region to clarify its action and mechanism. Two of the rabbits showed piloerection, anorexia, and somnolence, and died soon after booster administration. The serum calcium level of the dead rabbits was significantly high, compared to those of surviving rabbits. Interestingly, amino acid sequences homologous with gp46-197 were found in the carboxyl-terminal half of osteoprotegerin (OPG), an osteoclast inhibitory factor. To confirm the effect of the gp46-197 region on osteogenesis in vivo, the peptide was intraperitoneally administered to male Sprague-Dawley rats. The administration of the gp46-197 peptide resulted in a decrease of bone mineral density (BMD), a significant increase of serum calcium level, and inhibition of normal bone growth in both short- and long-term experiments. In rats, femoral growth inhibition by the gp46-197 peptide was restored by the coadministration of recombinant human OPG. Improvement by OPG in the adverse effect indicates that the central region of HTLV-1 Gp46 acts as an antagonist for OPG and leads to hypercalcemia. PMID- 19134005 TI - Inverse correlation of the up-regulation of FZD10 expression and the activation of beta-catenin in synchronous colorectal tumors. AB - We investigated the immunohistochemical expression patterns of Frizzled homolog 10 (FZD10), a cell-surface receptor for molecules in the Wnt pathway, in tissue samples derived from 104 patients with colorectal cancers (CRCs). There was no immunoreactivity for FZD10 in normal colonic mucosa, and only tumor cells in polyps and CRC tissues showed spotted immunostaining patterns in apical sides of the cytoplasm. In metastatic liver lesions, tumor cells showed cytoplasmic immunostaining similar to primary lesions, whereas normal liver parenchyma showed almost no immunostaining. Frequencies of FZD10-immunopositive cells in tumor tissues were significantly higher in CRCs than those in polyps (3.3 +/- 10.3% vs 20.5 +/- 31.7%, P = 0.0016), and almost equivalent with those in metastatic liver lesions (33.2 +/- 39.7% vs 26.4 +/- 33.4%, P = 0.133). Analyses of paired samples (polyps and CRCs, or CRCs and metastatic liver lesions from the same patient) suggested that a subset of CRCs possessed intrinsic genetic mechanisms causing the evolution of FZD10-positive clones during tumor progression, making FZD10 a promising candidate for molecular imaging and a target for therapy. To our surprise, cancer cells immunopositive for FZD10 showed significantly less nuclear accumulation of beta-catenin, compared to FZD10-immunonegative cancer cells, and there was a strong inverse correlation between nuclear immunostaining scores for beta-catenin expression and expression patterns of FZD10 (P = 0.0002), suggesting that FZD10 has a distinct role from other FZDs in canonical Wnt signal transduction. PMID- 19134007 TI - Transthyretin-driven oncolytic adenovirus suppresses tumor growth in orthotopic and ascites models of hepatocellular carcinoma. AB - Strategies to increase antitumor efficacy of oncolytic adenoviruses are actively investigated. We have previously shown that E1B-55 kDa-deleted adenovirus, designated Ad5WS1, has therapeutic potential for treating hepatocellular carcinoma (HCC). To achieve HCC-restricted replication of oncolytic adenovirus, we generated Ad5WS2, an E1B-55 kDa-deleted adenovirus with its E1A gene driven by the liver-specific transthyretin promoter. Our results showed that Ad5WS2 could replicate within tumor cells where the transthyretin gene was expressed. Mouse transthyretin promoter was active in murine and human HCC cells, but relatively quiescent in cells of non-liver origin. Ad5WS2 caused severe cytolytic effect on HCC cells, but was much attenuated in non-HCC cells. Peritoneal administration of Ad5WS2 into mice bearing liver tumors grown in ascites resulted in enhanced survival. In an orthotopic HCC model, Ad5WS2, when systemically administered, exerted higher antitumor effects than Ad5WS1. Lack of viral replication in normal organs and minimal hepatic toxicity was noted after Ad5WS2 treatment. Furthermore, the antitumor effect of Ad5WS2 could be enhanced when combined with chemotherapeutic agent cisplatin in the ascites tumor model. These results suggest that E1B-55 kDa-deleted adenovirus driven by the transthyretin promoter may be a safer and more efficacious oncolytic agent for the treatment of primary and metastatic HCC. PMID- 19134008 TI - Expression of Aurora B and alternative variant forms in hepatocellular carcinoma and adjacent tissue. AB - Surgical resection is the effective treatment modality for hepatocellular carcinoma (HCC); however, rapid recurrence of the tumors are frequently observed even after apparently curative resection. The recurrence and prognostic assessment of patients with HCC after resection is an important clinical issue. We recently reported that aberrant expression of Aurora B is observed in primary HCC, and that it can be a predictive factor for HCC recurrence exceeding Milan criteria after curative hepatectomy. In this study we investigated the expression of the newly observed Aurora B splicing variant forms in HCC, and their roles in hepatocarcinogenisis. The expression of Aurora B and splicing variant forms were screened in 125 HCC patients (94 chronic hepatitis with cirrhosis background liver specimens), 18 metastatic liver cancer patients and 16 normal liver specimens by cDNA microarray, reverse transcription -- polymerase chain reaction (RT-PCR) and Real time quantitative Reverse Transcription PCR (qRT-PCR). The results showed that expression of Aurora B splicing variant 2 (AURKB-Sv2) variant form was absent in normal liver and was higher in metastatic liver cancer than HCC. This aberrant expression was associated with the advanced stages of HCC (P < 0.01), correlated with a poor outcome (P = 0.008) and short disease-free period (P = 0.018). Furthermore, AURKB-Sv2 variant form is associated with a higher level of serum alpha-fetoprotein, protein induced by vitamin K absence or antagonist-II (PIVKAII), tumor capsular invasion, multiple tumor formation and at an age younger than those with other variant forms (P < 0.05). The results thus suggest that AURKB-Sv2 variant form is more significantly associated with the advanced stages of HCC than others and is a marker of poor prognosis. Founded in the tumor capsular invasion and multiple tumor regions, suggests that this might play a role in enhancing multiple malignant tumor formation and recurrence of HCC in hepatocarcinogenesis. This is the first study to report clinicopathological significance of aberrant expression of AURKB-Sv2 variant form in hepatocellular carcinoma. PMID- 19134009 TI - Parenting knowledge and its role in the prediction of dysfunctional parenting and disruptive child behaviour. AB - BACKGROUND: There is a paucity of research on the relationship between parental knowledge, parenting and parenting self-efficacy, and some inconsistencies have been reported in the literature. METHOD: Parent knowledge of effective parenting strategies was assessed among 68 parents from a non-clinic sample, who also completed questionnaires relating to parenting confidence, quality of parenting and child behaviour. RESULTS: Parents with greater knowledge tended to be less dysfunctional, and reported significantly higher education and income levels. Parenting confidence explained a significant proportion of the variance in reported frequency of disruptive child behaviour while knowledge did not independently contribute to the prediction. However, the relationship between parenting confidence and dysfunctional parenting was moderated by the level of knowledge. There was a stronger negative relationship between confidence and dysfunctional parenting when knowledge level was low than when it was high. Post hoc analyses indicated that the relationship between parenting knowledge and disruptive child behaviour was moderated by the level of parenting dysfunction. Parenting knowledge and reported frequency of disruptive behaviour were positively related when the level of dysfunction was low, but were unrelated when it was high. CONCLUSIONS: Parents with low levels of knowledge and confidence in their parenting may be at greater risk of dysfunctional parenting and might benefit from interventions designed to enhance both knowledge and confidence. Results are interpreted in relation to inconsistencies with previous research and implications for future methodologies. PMID- 19134010 TI - Quality of life, health satisfaction and family impact on caregivers of children with developmental delays. AB - OBJECTIVE: To study the quality of life, health satisfaction and family impact on caregivers of children with developmental delays in Taiwan. DESIGN: Cross sectional study. SUBJECTS: The caregivers of children with diagnoses of developmental delays recruited from a teaching hospital in northern Taiwan. METHODS: The main caregivers of 48 male and 22 female children with developmental delays were recruited. WHOQOL-BREF for health-related quality of life (HRQOL), PedsQL-Health Satisfaction for health satisfaction, PedsQL-Family Impact Module and Impact on Family Scale for family impact were evaluated. The correlation of caregivers' HRQOL, health satisfaction and family impact were also studied. RESULTS: Caregivers in nuclear families had higher health satisfaction scores (78.2 for nuclear families vs. 66.9 for extended families, P < 0.05) when assessed by the PedQL-Health Satisfaction questionnaire. Children's age was negatively correlated with family impact, including parent (-0.272, P = 0.023), family (-0.262, P = 0.029) and total scores (-0.281, P = 0.018) as assessed using the PedsQL-Family Impact Module. CONCLUSION: A negative relation between impact of burden and child's age suggests that family members gradually adapt to the delayed developmental status in their children as they grow. Caregivers in nuclear families having higher health satisfaction than those in extended families may be due to Chinese cultural effects. PMID- 19134011 TI - Self-perception in overweight and obese children: a cross-sectional study. AB - AIM: The aim of this study was to examine the relationship between obesity and self-esteem in children in relation to specific domains of their self-perception, and further to explore the extent to which this may vary by gender and economic circumstances. METHOD: A total of 211 children aged 8-9 years drawn from both advantaged and disadvantaged areas of Belfast completed the Harter Self Perception Profile for Children and measures of body mass index were obtained. RESULTS: Overweight, impoverished children had significantly reduced social acceptance and physical competence scores. Boys had significantly lower scores than girls in the behavioural conduct domain. Girls had significantly lower scores than boys for the athletic competence. CONCLUSION: These results suggest that risk factors of increased weight and impoverished backgrounds have a combined negative effect, placing some children at increased risk of having lower self-perceptions in some, but not all domains. Health interventions for childhood obesity should consider the likelihood of specific relationships between physical and psychosocial factors. PMID- 19134013 TI - Dog ownership might exert a protective effect on wheezing and atopy independently by endotoxin exposure. PMID- 19134014 TI - Functional analysis of a polymorphism in the promoter region of the IL-12/23p40 gene. AB - BACKGROUND: Human IL-12B gene on chromosome 5q31 encodes the common p40 subunit of IL-12 and IL-23. IL-12 is known to play critical roles in the generation of T helper type 1 (TH(1)) cells, whereas IL-23 is involved in maintenance and/or population expansion of TH(17) cells. Although several reports suggested an association between a polymorphism (-6415CTCTAA/GC) in IL-12B and asthma, the molecular mechanism how this polymorphism is involved in allergic inflammation is still unclear. METHODS: The transcription activity was analysed by reporter assay. A transcription factor binding to -6415 polymorphic site was identified by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay. The amount of cytokines produced from peripheral monocytes were determined by ELISA. RESULTS: Reporter assay showed that the transcription activity of the GC allele was higher than that of the CTCTAA allele. A transcription factor Sp1 bound to the region including the GC allele with a higher affinity than that of the CTCTAA allele in EMSA. In vivo binding of Sp1 to IL-12B gene carrying -6415GC was confirmed by ChIP assay. Overexpression of Sp1 up-regulated transcription activity of promoter carrying GC allele sequence, whereas the CTCTAA promoter was not affected by Sp1. We examined the correlation between -6415CTCTA/GC polymorphism and production of cytokine IL-12/23p40, IL 12p70, and IL-23 on peripheral blood monocytes, and monocytes with the GC/GC allele exhibited significantly higher expression of IL-12p70 protein than those with the CTCTAA/CTCTAA allele (P=0.009). CONCLUSIONS: The -6415 polymorphism is involved in cytokine production potential by affecting Sp1-mediated transcription activity. PMID- 19134015 TI - Role of Ca2+ mobilization and Ca2+ sensitization in 8-iso-PGF 2 alpha-induced contraction in airway smooth muscle. AB - BACKGROUND: Isoprostanes are prostaglandin (PG)-like compounds synthesized by oxidative stress, not by cyclooxygenase, and increase in bronchoalveolar lavage fluid of patients with asthma. The airway inflammation implicated in this disease may be amplified by oxidants. Although isoprostanes are useful biomarkers for oxidative stress, the action of these agents on airways has not been fully elucidated. OBJECTIVE: This study was designed to determine the intracellular mechanisms underlying the effects of oxidative stress on airway smooth muscle, focused on Ca(2+) signalling pathways involved in the effect of 8-iso-PGF(2 alpha). METHODS: Using simultaneous recording of isometric tension and F(340)/F(380) (an indicator of intracellular concentrations of Ca(2+), [Ca(2+)]i, we examined the correlation between tension and [Ca(2+)]i in response to 8-iso PGF(2 alpha) in the fura-2 loaded tracheal smooth muscle. RESULTS: Augmented tension and F(340)/F(380) by 8-iso-PGF(2 alpha) were attenuated by ICI-192605, an antagonist of thromboxane A(2) receptors (TP receptors). Moreover, D609, an antagonist of phosphatidylcholine-specific phospholipase C, markedly reduced both the tension and F(340)/F(380) induced by 8-iso-PGF(2 alpha), whereas U73122, an antagonist of phosphatidylinositol-specific phospholipase C, modestly inhibited them by 8-iso-PGF(2 alpha). SKF96365, a non-selective antagonist of Ca(2+) channels, markedly reduced both tension and F(340)/F(380) by 8-iso-PGF(2 alpha). However, diltiazem and verapamil, voltage-dependent Ca(2+) channel inhibitors, modestly attenuated tension although their reduction of F(340)/F(380) was not different from that by SKF96365. Y-27632, an inhibitor of Rho-kinase, significantly attenuated contraction induced by 8-iso-PGF(2 alpha) without reducing F(340)/F(380), whereas GF109203X and Go6983, protein kinase C inhibitors, did not markedly antagonize them although reducing F(340)/F(380) with a potency similar to Y-27632. CONCLUSION: 8-iso-PGF(2 alpha) causes airway smooth muscle contraction via activation of TP receptors. Ca(2+) mobilization by SKF96365- and D609-sensitive Ca(2+) influx and Ca(2+) sensitization by Rho-kinase contribute to the intracellular mechanisms underlying the action of 8-iso-PGF(2 alpha). Rho-kinase may be a therapeutic target for the physiologic abnormalities induced by oxidative stress in airways. PMID- 19134016 TI - Efficacy and safety of high-doses sublingual immunotherapy in ultra-rush scheme in children allergic to grass pollen. AB - BACKGROUND: Although sublingual immunotherapy (SLIT) has been used with increasing frequency, the data on the efficacy of SLIT in pediatric asthma are limited. AIM: The aim of our study was to evaluate the efficacy and the safety of high-dose SLIT given pre-seasonally and co-seasonally in an ultra-rush scheme in children with bronchial asthma allergic to grass pollen. METHODS: Fifty children with asthma, aged 6-17, sensitive to grass pollen, participated in the 2-year prospective, randomized, double-blind, placebo-controlled trial, to investigate the efficacy and safety of SLIT (Staloral 300 IR, Stallergenes SA, 25 microg major allergens) as a standardized extract of five grass pollen with ultra-rush induction. RESULTS: SLIT significantly improved asthma symptom scores (41% vs. placebo group), reduced nasal symptoms (25% vs. placebo group) and the use of rescue medications (10% vs. placebo group), improved forced expiratory volume in 1 s, but had no effect on ocular symptoms, nasal hyper-reactivity, peak expiratory flow and forced expiratory volume between 25% and 75% of vital capacity. Serum levels of immunoglobulin E and IgG4 did not change after SLIT. After the second season of SLIT, an improvement in bronchial hyperresponsiveness was observed; however, compared with placebo, this effect was not significant. Among all subjects in SLIT group, predominantly local reactions have been recorded in 59% of subjects in the first year of treatment and in 35% in the second. CONCLUSIONS: Our study indicated that high-dose ultra-rush, co-seasonal SLIT given for 2 years, was safe and reduced a multiple symptom-medication score. PMID- 19134017 TI - CD200R surface expression as a marker of murine basophil activation. AB - BACKGROUND: Basophils are increasingly recognized as playing important roles in the immune responses of allergic diseases and helminth infections. One of the main obstacles to studying basophils has been the lack of a simple and rapid assay to measure basophil activation in mice. OBJECTIVE: The purpose of this study was to develop an assay to measure murine basophil activation. METHODS: Mouse blood cells were stained with various combinations of positive and negative markers for basophils--sorted and then assessed for basophil purity by May Grunwald staining of cytospins. Once a flow cytometric strategy for staining basophils was determined, basophil surface expression of CD200R was assessed by multi-colour flow cytometry after stimulation of whole blood with anti-IgE, ionomycin or N-formyl MetLeuPhe (fMLP). Confirmation of basophil activation was assessed by concomitant staining of cells for intracellular IL-4. To test the ability of flow cytometric basophil CD200R measurements to assess for antigen specific IgE-mediated activation of basophils, surface CD200R expression in response to in vitro stimulation with media alone, helminth antigen or ovalbumin was measured on basophils obtained from control mice, mice infected with helminths and mice sensitized to ovalbumin. RESULTS: Using anti-IgE-FITC as a positive marker and a combination of anti-CD4-PERCP and anti-B220-PERCP as negative markers resulted in a well-separated basophil population. Additional staining with anti-CD200R-PE demonstrated that (1) basophil CD200R expression increases in response to anti-IgE, ionomycin and fMLP, (2) most CD200R-positive basophils also stain positively for IL-4 and (3) CD200R expression increases after antigen-specific activation of basophils in murine models of helminth disease and allergy. CONCLUSION: We developed a multi-colour flow cytometry assay that measures murine basophil activation by utilizing CD200R as an activation marker. This assay is straightforward and rapid, taking approximately half a day for obtaining blood, in vitro stimulation and flow cytometric analysis. PMID- 19134018 TI - Implementation of pre-seasonal sublingual immunotherapy with a five-grass pollen tablet during optimal dosage assessment. AB - BACKGROUND: The optimal dose (300IR) of a five-grass pollen sublingual immunotherapy tablet in terms of efficacy was previously demonstrated from the first pollen season. OBJECTIVE: Here, we aim to confirm whether this dose remained optimal during the peak of the pollen season by assessing the efficacy and quality of life data. METHODS: A total of 628 subjects with grass pollen rhinoconjunctivitis were randomized in a double-blind, placebo-controlled, multi centre, pan-European trial. Subjects received once-daily tablets (Stallergenes, Antony, France) of 100IR, 300IR, 500IR or placebo, starting 4 months before and throughout the 2005 grass pollen season. The pollen season was defined as the first day of 3 consecutive days with a grass pollen count above 30 grains/m(3) of air, recorded using Hirst-type volumetric pollen traps, to the last day before 3 consecutive days with a pollen count below 30 grains/m(3). RESULTS: The grass pollen season lasted an average of 30 days, with a peak of 12 days. The mean treatment duration before the grass pollen season was similar in the four treatment groups (121.4+/-31.1 to 128.6+/-15.4 days in the safety population). Both the 300IR and 500IR groups had highly significant improvements in Rhinoconjunctivitis Total Symptom Score (RTSS) vs. placebo at the peak pollen season (P=0.0005 and 0.0014, respectively), which agreed with improvements in RTSS in the primary evaluations. The average RTSS scores were slightly elevated during the peak pollen season in all treatment groups. The overall Rhinoconjunctivitis Quality of Life Questionnaire score confirmed the optimal dosage 300IR at peak (P<0.0001) and at the end (P< or =0.0031) of the pollen season. All doses were well tolerated. CONCLUSION: At the peak pollen season, the efficacy and quality of life data for both 300IR and 500IR groups was significantly improved vs. the placebo group. These results confirm the conclusions of the primary evaluations and validate the use of 300IR tablets for clinical practice. PMID- 19134019 TI - Efficacy and safety of 5-grass pollen sublingual immunotherapy tablets in patients with different clinical profiles of allergic rhinoconjunctivitis. AB - BACKGROUND: The optimal dose of grass pollen tablets for sublingual immunotherapy (SLIT) in allergic rhinoconjunctivitis patients was previously established in a multinational, randomized, double-blind, placebo-controlled study in 628 adults. Patients were randomized to receive once-daily 5-grass pollen sublingual tablets of 100 IR (index of reactivity), 300 IR or 500 IR, or placebo starting 4 months before the pollen season. OBJECTIVE: The aim of this complementary analysis was to determine whether 300 IR 5-grass pollen SLIT-tablets is effective in different subtypes of patients who are allergic to grass pollen. METHODS: Different subgroups could be identified regarding comorbidities (with or without asthma during the grass-pollen season), sensitization (mono/polysensitization) and symptom severity. An additional exploratory analysis was performed within four subgroups based on pre-treatment assessment: Group 1=high specific IgE; Group 2=high symptom scores; Group 3=high skin sensitivity; Group 4=any of Group 1, 2 or 3. RESULTS: Asthma and sensitization status were not significant covariates as the average Rhinoconjunctivitis Total Symptom Score (RTSS) was identical for patients with and without grass-pollen asthma, as well as for mono- and polysensitized patients. Across the four subgroups, average RTSSs (+/- SD) for the optimal dosage (300 IR) were 3.91 +/- 3.16, 3.83 +/- 3.14, 2.55 +/- 2.13 and 3.61 +/- 2.97, for subgroups 1, 2, 3 and 4, respectively. ANCOVA showed that in Group 1 average RTSS did not differ significantly with different doses of SLIT. In Groups 2, 3 and 4, doses of 300 IR and 500 IR were significantly more effective than 100 IR and placebo (P< or =0.035). All doses of SLIT administered in this study can be considered safe in the patients investigated. CONCLUSIONS: The risk-benefit ratio validates the use of 300 IR tablets in clinical practice in all of these patient subgroups, regardless of severity profile, sensitization status and presence of asthma. PMID- 19134020 TI - Probiotic supplementation in the first 6 months of life in at risk Asian infants- effects on eczema and atopic sensitization at the age of 1 year. AB - BACKGROUND: The role of probiotics in allergy prevention remains uncertain but has been shown in some studies to have a possible protective effect on eczema. OBJECTIVE: We aimed to assess the effect of probiotic supplementation in the first 6 months of life on eczema and allergic sensitization at 1 year of age in Asian infants at risk of allergic disease. METHODS: A double-blind, placebo controlled randomized clinical trial involving 253 infants with a family history of allergic disease was carried out. Infants received at least 60 mL of commercially available cow's milk formula with or without probiotic supplementation [Bifidobacterium longum (BL999) 1 x 10(7) colony forming unit (CFU)/g and Lactobacillus rhamnosus (LPR) 2 x 10(7) CFU/g] daily for the first 6 months. Clinical evaluation was performed at 1, 3, 6 and 12 months of age, with serum total IgE measurement and skin prick tests conducted at the 12-month visit. The primary and secondary end-points were eczema and allergen sensitization, respectively. RESULTS: The incidence of eczema in the probiotic (22%) group was similar to that in the placebo group (25%) (P=0.53). The median Scoring Atopic Dermatitis score at 12 months was 17.10 (9.74) in the probiotic group and 11.60 (8.40) in the placebo group (P=0.17). The prevalence of allergen sensitization showed no difference (probiotic=24% vs. placebo=19%, P=0.26). The total IgE geometric mean (95% confidence interval) was 18.76 (12.54-24.98) kU/L in the probiotic group and 23.13 (16.01-30.24) kU/L in the placebo group (P=0.15). Atopic eczema (with sensitization) in the probiotic (7.3%) group was comparable to the placebo group (5.8%) (P=0.86). CONCLUSION: Early life administration of a cow's milk formula supplemented with probiotics showed no effect on prevention of eczema or allergen sensitization in the first year of life in Asian infants at risk of allergic disease. Further work is needed to determine whether timing of supplementation, dose and probiotic strain are important considerations. PMID- 19134021 TI - Detection of allergen composition and in vivo immunogenicity of depigmented allergoids of Betula alba. AB - BACKGROUND: Chemical modification of allergen vaccines to reduce IgE binding improves safety while maintaining clinical efficacy. However, this also complicates the characterization of allergoids using techniques as for native allergen extracts. OBJECTIVES: The objective of this study was to analyse the molecular size of Betula alba depigmented allergoids, conservation of major allergens in the allergoids and in vivo antibody response to immunization. METHODS: The molecular size of depigmented allergoids was evaluated by high performance-size exclusion chromatography and light scattering techniques. Protein composition was compared with native extracts by capillary liquid chromatography-tandem mass spectrometry based peptide mapping. Rabbits were immunized with depigmented allergoid of Betula pollen adsorbed onto aluminium hydroxide (Depigoid). IgG antibodies against individual allergens were determined by ELISA and immunoblot. RESULTS: Depigmented allergoids contained a range of high molecular weight particles, approximately 60% of which had a molecular weight of 1-3 MDa. Peptide sequencing confirmed the preservation of five isoforms of Bet v 1, as well as Bet v 2, Bet v 6 and Bet v 7. Sera from immunized rabbits showed high levels of specific IgG to rBet v 1.0101 and rBet v 2. CONCLUSIONS: The mean protein content was 544+/-106 microg per mg of freeze-dried material for depigmented allergoids and 434+/-71 for native extracts. They retain the capacity to induce specific IgG antibodies against individual allergens present in the native extract. These findings confirm the immunogenicity of depigmented allergoids and may explain why patients treated with these vaccines are protected against the native allergens. Analysis of molecular size and allergen content may be useful techniques for characterization and standardization of allergoid products. PMID- 19134022 TI - First evidence of a possible association between gastric acid suppression during pregnancy and childhood asthma: a population-based register study. AB - BACKGROUND: Recent data in mice suggest that acid suppression during pregnancy yields offspring with type 2 T helper-dominant immunity, suggesting a predisposition for allergy. OBJECTIVE: To determine the association of in utero exposure to acid-suppressive medications and the subsequent development of allergic diseases in children. METHODS: We studied a population-based observational cohort formed by linking data from three Swedish national healthcare registers: the Medical Birth Register, the Hospital Discharge Register, and the Swedish Prescribed Drug Register. Main outcome measures included a hospital discharge diagnosis of an allergic disease or prescription for asthma medications, epinephrine auto-injectors, antihistamines or steroid ointments in children. Data were analysed using the Mantel-Haenszel procedure. RESULTS: Twenty-nine thousand four hundred and ninety (5.03%) children had a discharge diagnosis of allergy or prescriptions for allergy medications. Five thousand six hundred and forty-five (0.96%) children had been exposed to acid suppression therapy during pregnancy; of these, 405 (0.07%) were treated for allergic diseases. Exposure to acid-suppressive medications in utero was associated with an increased odds ratio (OR) for developing allergy (OR 1.43, 95% confidence interval (95% CI) 1.29-1.59). We observed this association irrespective of the type of drug, time of exposure during pregnancy, and maternal history of allergy. The use of maternal acid-suppressive medication was associated with an increased OR for the development of childhood asthma (3.7% in the population at large vs. 5.6% in exposed children, OR 1.51, 95% CI 1.35-1.69), but not for other allergic diseases. CONCLUSION: These data provide first evidence of a significant association between in utero exposure to acid suppressive drugs and the risk of developing childhood asthma. PMID- 19134023 TI - AML transformation in 56 patients with Ph- MPD in two well defined populations. AB - The Philadelphia chromosome-negative (Ph-) chronic myeloproliferative disorders (MPD) have an inherent tendency for transformation into acute myelogenous leukaemia (AML). The long-term rate of leukaemic transformation in unselected MPD patients was studied in well-defined MPD populations in Gothenburg, Sweden and the Cote d'Or area, Burgundy, France, respectively. Over a median observation time of 15 yr, 56 subjects (7%) out of a total of 795 patients with Ph- MPD transformed to AML. The yearly incidence of AML transformation was 0.38% in polycythaemia vera (PV), 0.37% in essential thrombocythaemia (ET) and 1.09% in idiopathic myelofibrosis (IMF). The incidence of AML development was significantly higher in IMF as compared with both PV and ET (P = 0.002 and P = 0.02, respectively). Six of the patients who developed AML had never been treated with cytoreductive agents and two had only been exposed to interferon. In IMF, the average time from diagnosis to AML transformation was 42 +/- 33 months, which was significantly shorter than for both PV and ET (88 +/- 56 and 76 +/- 57 months; P = 0.0075 and P = 0.027, respectively). The time from diagnosis to AML transformation appears to be a continuous event as regards all three MPD entities. It was shown that 17 out of the 18 patients with PV who developed AML were females; this was true despite the fact that the male/female ratio for the whole PV group was 146/171 (0.85). As regards ET and IMF patients who transformed to AML, the gender ratio showed slight male predominance (1.33 and 1.13, respectively). The average survival time for the 56 MPD patients who developed AML was 4.6 +/- 5.5 (range 0-28) months and did not differ with respect to the three subtypes of pre-AML MPD. PMID- 19134024 TI - Prompt recovery of chemotherapy associated Hand-Foot syndrome treated with acetylsalicylic acid in two patients with Hodgkin's lymphoma. PMID- 19134025 TI - Prevention of haemophilic arthropathy in haemophilic children with inhibitors. AB - Prophylaxis is the ideal manner to treat haemophilia patients without inhibitors. In fact, it can prevent the orthopaedic complications that commonly bedevil haemophilia patients without inhibitors in the absence of such prophylaxis or when they are only managed by on-demand treatment. Currently, there are two agents that allow us to control haemostasis in patients with inhibitors, i.e. recombinant factor VIIa (rFVIIa) and activated prothrombin complex concentrate (APCC). Both agents can be used for arthropathy prevention, or for intensive on demand treatment in haemophilia patients with inhibitors. In this paper, we have tried to find an answer to the question of which bypassing agents (rFVIIa or APCC) could be used in children and adults. PMID- 19134026 TI - Haemophilic arthropathy: the importance of the earliest haemarthroses and consequences for treatment. AB - In a case of massive bleeding, the possibility of puncturing the joint can be considered to reduce the total load of intra-articular blood, and thus the total load of iron which, in time, can be found in the synovium with devastating long term effects. Subsequent initiation of prophylaxis, still very early in life, might be more beneficial for the preservation of joints. Thus, it might be argued that the initiation of primary prophylaxis should be based on joint haemorrhage history rather than age and, according to some authors, preferably after the bleed in a single joint. The high cost of recombinant factor VIII may make widespread acceptance of prophylaxis impractical. Thus, beneficial results should be balanced with cost considerations. In this balance of treatment cost and efficacy, it must be taken into account that improperly treated haemophilia patients make great demands on healthcare systems (in terms of costs) because of their need for additional treatments such as expensive joint replacement surgery. However, where resources are limited, it can be argued that prophylaxis may be stopped in adulthood, in a certain proportion of all adult patients, with acceptable consequences for orthopaedic outcome in the long term. By doing so, limited amounts of clotting factor can be used for young patients with optimal cost effectiveness. PMID- 19134027 TI - Prevention of arthropathy in haemophilia - intensive on-demand treatment, UK perspective. AB - In the past, there were only minimal studies aimed at establishing the dose of concentrate required for treating an acute joint bleed in patients without inhibitors. The cost of treatment with bypassing agents consumes huge economic resources and therefore it is important to have rigorous studies to establish rational dosing. Both the FEIBA NovoSeven Comparative study and the comparative dosing study have been bold attempts to clarify treatment regimens for managing joint bleeds in patients with inhibitors. As a very challenging situation in which to conduct research, it is important that such endeavour continues. PMID- 19134028 TI - Prevention of arthropathy in haemophilia: prophylaxis. AB - At present, when resources allow, the main issue in preventing arthropathy in patients with haemophilia is determining the 'optimal' prophylaxis regimen rather than the choice between primary prophylaxis and on-demand treatment. Barriers to prophylaxis implementation are still relevant and include difficulties with venous access, adherence to treatment regimens and costs. Prospective studies with different prophylactic regimens and objective outcome measures are still needed to determine the best cost-benefit ratio. The results from large retrospective cohorts followed up for decades in several European countries have shown that regular prophylaxis started at an early age is able to reduce physical impairment from haemophilic arthropathy. The efficacy of primary prophylaxis was finally demonstrated also in the setting of a randomized clinical trial. PMID- 19134029 TI - Articular status of haemophilia patients with inhibitors. AB - The incidence of haemarthrosis is similar in young haemophilia patients with or without inhibitors, but whether this is due to the effect of modern therapy is unknown. In comparison with younger patients with haemophilia, older haemophilia patients have a lower incidence of haemarthrosis because of very severe arthropathy (crippled joints). It is not known whether this could be due to the lack of therapy in previous years. In addition, the burden of orthopaedic complications and the impact on quality of life are more severe in patients with haemophilia who have developed an inhibitor compared with those without inhibitors. This article describes the results of the European Study on Orthopaedic Status of Haemophilia Patients with inhibitors (ESOS) which set out to determine how the inhibitor status of haemophilia patients may influence their quality of life and degree of arthropathy. PMID- 19134030 TI - Prevention of arthropathy: may it be extrapolated from patients without inhibitors to patients with inhibitors? AB - Although no consensus on the exact dosing schedule of prophylaxis in non inhibitor haemophilia patients has been achieved, regular administration of factor VIII (FVIII) or factor IX (FIX) concentrates is currently accepted as the most effective treatment model in order to prevent sick days, hospital stays and the development of arthropathy in patients with severe haemophilia. The basis for prophylaxis has been the idea to increase the plasma levels of FVIII or FIX above 1% thereby improving the haemostasis in severe haemophilia. Current treatment in haemophilia patients with inhibitors includes the use of activated prothrombin complex concentrates and recombinant FVIIa (rFVIIa). Regular administration of such agents has recently repeatedly been reported to decrease the number of joint and muscle bleeds in these patients. Regarding rFVIIa, such an effect was confirmed in a controlled study published in 2007. Although the exact mechanism of action of rFVIIa in this setting is still not clear, it seems reasonable to assume that prophylaxis with rFVIIa will be able to prevent or minimize the development of haemophilic arthropathy. As arthropathy is a long-term complication in haemophilia and the result of repeated joint bleeds, a longer follow-up of patients on regular prophylaxis including measurements of the joint scores, as has been performed in FVIII/FIX prophylaxis in non-inhibitor patients, will be required for a definite conclusion. PMID- 19134031 TI - Experiences in the prevention of arthropathy in haemophila patients with inhibitors. AB - Haemophilia patients with inhibitor have a higher level of arthropathy and more severe joint morbidity than patients without inhibitors. In recent years, interest has grown in the possibility that bypassing agent regimens could prevent bleeding and, consequently, arthropathy in inhibitor patients. Nevertheless, doubts about efficacy, complications and cost exist, questioning the justification of an uncertain prophylaxis in patients with inhibitors. Activated prothrombin complex concentrate (aPCC) has been used in more than 70 haemophilia patients with inhibitors in different clinical situations. aPCC prophylaxis seems to be safe and effective for the reduction of bleeding episodes in some patients. Recombinant activated factor VII (rFVIIa) has been employed prophylactically in over 44 haemophilia patients with inhibitors; 22 patients were included in the only randomized, prospective clinical trial of bypassing agents in prophylaxis. Bleeding frequency was reduced and this reduction was maintained during the postprophylaxis period. No thromboembolic events were reported during prophylaxis with rFVIIa. Although the effect of aPCC can last longer than that of rFVIIa, their efficacy rates are similar, suggesting that the biological effect of rFVIIa is actually much longer than indicated by its short plasma half-life. aPCC contains residual factor VIII antigen and may cause an anamnestic response in the inhibitor titre. This is crucial when immune tolerance induction is postponed to allow the inhibitor titre to decline to <10 Bethesda Units. In this setting, aPCC is not recommended as a first-line prophylaxis because of its potential to protract anamnesis, and rFVIIa is the preferred agent. PMID- 19134033 TI - The role of rehabilitation and sports in haemophilia patients with inhibitors. AB - Treatment of haemophilia patients with inhibitors against factor VIII/IX (FVIII/IX) is still challenging and recurrent haemarthroses cause arthropathy with associated restrictions on participation in physical activities and sports. Rehabilitation is a multidisciplinary approach which includes physiotherapy, occupational therapy, psychology, social work and technical applications like prostheses, orthoses (splints and braces), shoe adaptations, walking aids and adaptations in the house and work situation, but also education. The theoretical principles and practical advice regarding rehabilitation and physiotherapy for both children and adults with haemophilia without inhibitors are highly applicable for patients with inhibitors. Hydrotherapy is useful in the treatment of painful or stiff joints and/or muscles after an acute haemarthrosis, muscle bleeds and chronic arthropathy. In addition, it is of use in cases of chronic synovitis and to start mobilization after long periods of bed rest or during the weaning of a splint. In cases of bleeding and arthropathy, adequate treatment of pain is very important, as are functional exercises. Everyone should be physically active for 30-60 min day(-1). Participation in sports is recommended for people with haemophilia, the best sport being swimming. Children should participate in sports appropriate to their size and physical characteristics. PMID- 19134032 TI - Clinical and echographical control protocol of haemarthrosis in haemophilia patients with inhibitors: evaluation of the efficacy of recombinant factor VIIa in the evolution process (EFFISEVEN protocol). AB - The intention of the 'clinical and echographical protocol of evaluation the efficacy of recombinant activated factor VII in the haemarthrosis' (EFFISEVEN protocol) was to provide an extensive study of the evolution of haemarthrosis, and second, of its repercussions on the degenerative process of joints. The clinical evaluation of haemarthroses and their evolution is based on a well established methodology, although very few studies have sought to determine the correlation between pain, mobility and the objective data regarding the haemorrhage. We believe that it is necessary to unify criteria and that the EFFISEVEN protocol may contribute data that improve standards which, in turn, will influence the degenerative process of joints, and consequently affect the quality of life of haemophilia patients with inhibitors. Echographical control of haemarthrosis is an objective method that allows control over how the haemorrhage evolves and also helps in the identification of rebleeding. Therefore, its use adds a new dimension to patient management strategies. Techniques used to monitor recombinant activated factor VII (rFVIIa) treatment require further study, although preliminary results guarantee their efficacy. PMID- 19134034 TI - Synoviorthesis in haemophilia patients with inhibitors. AB - Recurrent bleeding into joints represents the clinical hallmark of haemophilia and, if not adequately treated, it may cause chronic synovitis and degenerative arthropathy. In haemophilia patients with inhibitors, a more severe degree of synovitis is often observed owing to the fact that treatment is more problematic in this setting. The first treatment option of recurrent haemarthroses and/or chronic synovitis is represented by synoviorthesis, both chemical and radioisotopic, with a success rate of approximately 80% for both. However, radioisotopic synoviorthesis should be preferred in inhibitor patients because it makes it possible to obtain complete synovial fibrosis usually in one session, without the need for repeated injections, thus reducing the risk of bleeding complications and concentrate consumption. For all these reasons this procedure should be implemented and supported, particularly in developing countries. PMID- 19134035 TI - Orthopaedic surgery in haemophilia patients with inhibitors as the last resort. AB - Haemophilia patients with inhibitors commonly present much more severe musculoskeletal problems than patients without inhibitors. With haematological advances using recombinant activated factor VII (rFVIIa) and activated prothrombin complex concentrate (aPCC), it is now possible to perform orthopaedic procedures in these patients with a high rate of success. In Spain, the healthcare system sets no financial limits for the treatment of haemophilia, and children with haemophilia today have a much better musculoskeletal status than 30 years ago. However, the children of 30 years ago are now adults and are afflicted by severe articular deformities that frequently require the intervention of an orthopaedic surgeon. Our view is that the well-known concepts of prophylaxis in patients without inhibitors should be followed and applied to patients with inhibitors. In fact, prophylaxis seems to be the only way to prevent the development of haemophilic arthropathy in patients with haemophilia. Until this goal can be achieved, major and minor orthopaedic procedures will be necessary in patients with inhibitors more frequently than in patients without inhibitors. PMID- 19134036 TI - Prevention of haemophilic arthropathy during childhood. May common orthopaedic management be extrapolated from patients without inhibitors to patients with inhibitors? AB - We recommend prophylaxis in haemophilic children with an inhibitor as a way of preventing the musculoskeletal impairment that is likely to affect them. This approach has been used for children without inhibitors with excellent results. If prophylaxis is not feasible, we suggest that intensive on-demand treatment should be given. Two agents, recombinant activated FVII (rFVIIa) and activated prothrombin complex concentrates (aPCC), are currently used to control haemostasis either for prophylaxis or intensive on-demand treatment. As it is recombinant, rFVIIa would seem more appropriate to be employed in children. aPCC could be used in adults, or in the event of an unsatisfactory response to rFVIIa. We recommend prophylaxis or, at least, intensive on-demand treatment in haemophilia children with inhibitors. Both rFVIIa and aPCC are being used for this purpose. It would seem that rFVIIa might be more appropriate for children as it is a recombinant product. Nevertheless, after skeletal maturity (in adults), both agents could be used indistinctively (taking into consideration that FEIBA is a plasma-derived product). We still need more well-designed comparative studies in order to be able to assert that our consensus-based conclusion is evidence based. In orthopaedic surgery, both aPCC and rFVIIa have been reported to be effective in controlling perioperative haemostasis, although in practice most centres have so far used rFVIIa for their orthopaedic procedures. We recommend rehabilitation programmes for all patients with inhibitors in order to mitigate the disabling and handicapping impact of their condition and thereby enable them to achieve social integration. Programmes for haemophilic children without inhibitors can be applied to children with inhibitors but should be individually tailored. PMID- 19134037 TI - Incidence and factors related to flare-ups in a graduate endodontic programme. AB - AIM: To investigate the incidence and factors related to endodontic flare-ups in nonsurgical root canal treatment (NSRCT) cases completed by graduate endodontic residents at University of Pennsylvania, USA. METHODOLOGY: Residents at University of Pennsylvania enter all clinical patient records into an electronic database called PennEndo database. Analysis of records of 6580 patients treated from September 2000 to July 2005 revealed a total of 26 patients with flare-ups (0.39%). Patients were categorized to have undergone flare-up when they attended for an unscheduled visit and active treatment, and when they suffered from severe pain and or swelling after initiation or continuation of NSRCT. SAS software was used to develop a logistic regression model with flare-up as a dependent variable. Independent variables included in the model were: history of previous pain, one vs. two visit NSRCT, periapical diagnosis, tooth type, rotary versus hand instrumentation, and lateral versus vertical compaction of gutta-percha. RESULTS: The odds for developing a flare-up in teeth with a periapical radiolucency were 9.64 times greater than teeth without a periapical radiolucency (P = 0.0090). There was no statistically significant difference in flare-ups between one and two visits NSRCT. The odds of developing a flare-up increased 40 fold when NSRCT was completed in three or more visits. However, this result may have been confounded by addition of an unscheduled visit in patients suffering from flare-ups. Other independent variables did not have any statistically significant correlations. CONCLUSIONS: A low percentage of patients experienced flare-ups during NSRCT procedures. The presence of a periapical lesion was the single most important predictor of flare-ups during NSRCT. PMID- 19134038 TI - Randomized clinical trial of root-end resection followed by root-end filling with mineral trioxide aggregate or smoothing of the orthograde gutta-percha root filling--1-year follow-up. AB - AIM: To compare healing after root-end resection with a root-end filling of mineral trioxide aggregate (MTA) or smoothing of the orthograde gutta-percha (GP) root filling. METHODOLOGY: Forty-four patients (consisting of 52 teeth with periapical infection), average age of 54.6 years (range 30-77) participated in a randomized clinical trial (RCT) comparing the MTA and GP treatment methods. Radiographs produced 1-week and 12 months post-operatively were compared after blinding for treatment method, and healing was assessed as complete, incomplete, uncertain, or unsatisfactory. RESULTS: Six teeth were not available for the 12 month follow-up: three teeth (GP) had been re-operated because of pain and two teeth (one GP, one MTA) had been extracted because of root fracture (these five teeth were classified as failures). One patient (GP) was not available for recall. In the GP group, seven teeth (28%) showed complete healing, six teeth (24%) incomplete healing, six teeth (24%) uncertain healing and two teeth (8%) unsatisfactory healing after 1 year. In the MTA group, 22 teeth (85%) showed complete healing, three teeth (12%) incomplete healing, and none were scored as uncertain or unsatisfactory healing after 1 year. The difference in healing between the GP and the MTA groups was significant (P < 0.001). CONCLUSIONS: The results from this RCT emphasize the importance of placing a root-end filling after root-end resection. Teeth treated with MTA had significantly better healing (96%) than teeth treated by smoothing of the orthograde GP root filling only (52%). PMID- 19134039 TI - Accuracy of an electronic apex locator in primary teeth with root resorption. AB - AIM: To evaluate whether root resorption of primary molar teeth affects the accuracy of the Root ZX apex locator and to compare the Root ZX at different meter readings with direct canal measurement. METHODOLOGY: Sixty extracted primary molar teeth with root resorption affecting one sixth to one third of root length were used. The teeth were embedded in an alginate model. A K-type file was used in association with a Root ZX apex locator to measure canal length. Measurements were recorded using the Root ZX meter reading 'Apex' and '0.5 bar'. Actual tooth length was measured with a K-file to the major foramen. All measurements were read under a stereomicroscope at 15x magnification. The deviation of the Root ZX measurement from the actual canal length was determined. RESULTS: Mean differences between Root ZX length meter reading 'Apex' and actual length were 0.01 +/- 0.23 mm whereas mean differences between Root ZX length meter reading '0.5 bar' and actual length were -0.33 +/- 0.30 mm. The Root ZX was 96.7% accurate to within +/- 0.5 mm of the apical foramen when compared with the actual canal length of primary molars with root resorption. CONCLUSIONS: Using a criterion of +/- 0.5 mm, the accuracy of the Root ZX was high and not affected by root resorption. When compared with direct canal measurement, the error in locating the apical foramen was smaller with measurement at meter reading 'Apex' than meter reading '0.5 bar'. PMID- 19134040 TI - Evaluation of two mineral trioxide aggregate compounds as pulp-capping agents in human teeth. AB - AIM: The present randomized, controlled prospective study evaluated the histomorphological response of human dental pulps capped with two grey mineral trioxide aggregate (MTA) compounds. METHODOLOGY: Pulp exposures were performed on the occlusal floor of 40 human permanent pre-molars. The pulp was capped either with ProRoot (Dentsply) or MTA-Angelus (Angelus) and restored with zinc oxide eugenol cement. After 30 and 60 days, teeth were extracted and processed for histological examination and the effects on the pulp were scored. The data were subjected to Kruskal-Wallis and Conover tests (alpha = 0.05). RESULTS: In five out of the 40 teeth bacteria were present in pulp tissue. No significant difference was observed between the two materials (P > 0.05) in terms of overall histological features (hard tissue bridge, inflammatory response, giant cells and particles of capping materials). Overall, 94% and 88% of the specimens capped with MTA-Angelus and ProRoot, respectively, showed either total or partial hard tissue bridge formation (P > 0.05). CONCLUSIONS: Both commercial materials ProRoot (Dentsply) and MTA-Angelus (Angelus) produced similar responses in the pulp when used for pulp capping in intact, caries-free teeth. PMID- 19134041 TI - Effect of ethanol application on post-luting to intraradicular dentine. AB - AIM: To examine the effect of the application of an ethanol rinse before luting fibre posts to intraradicular dentine with etch-and-rinse adhesive systems by means of push-out bond strength evaluation and scanning electron microscopic (SEM) analysis. METHODOLOGY: Fibre posts were luted to single-canal premolars using Dual Link as a luting agent in combination with a three-step (All Bond 2) or a two-step (One Step Plus) etch-and-rinse adhesive system, which were applied as per manufacturers' instructions (control) or with the additional rinse of ethanol on acid-etched dentine prior to the bonding procedures (experimental). Bonded specimens were sectioned into 1-mm-thick slabs and subjected to push-out bond strength testing. In addition, specimens from each group were processed for SEM analysis. Data were analysed by Kruskal-Wallis followed by Dunn's post hoc test at P = 0.05. RESULTS: Push-out bond strength of All Bond 2 was significantly increased if the adhesive was applied on ethanol-saturated dentine (P < 0.05), whilst no significant difference (P > 0.05) was detected amongst experimental and control groups for One Step Plus. Irrespective of the adhesive, the SEM analysis revealed good impregnation patterns when both bonding techniques were employed. CONCLUSIONS: The use of the additional ethanol rinse on acid-etched dentine revealed higher bond strength of All Bond 2 when compared with the control application procedure, used in combination with a resin-based cement to lute fibre posts into the dowel space. However, no bond strength improvements were detected using One Step Plus. Further investigations are needed to develop a clinically applicable ethanol/bonding/luting technique. PMID- 19134042 TI - The effect of resin-based sealers on fracture properties of dentine. AB - AIM: To determine whether resin-based sealer cements are able to strengthen root dentine, as measured by work of fracture (Wf), micro-punch shear strength (MPSS) and resistance to vertical root fracture (VRF). METHODOLOGY: One hundred and twenty extracted premolar teeth were randomly assigned amongst four treatments before testing: intact, root canals prepared but unfilled, or root filled using epoxy- or urethane dimethacrylate (UDMA)-based sealer (plus core material). Samples were then prepared for measuring Wf, MPSS or VRF using standard test procedures. Data were analyzed using one-way anova with significance set at P < 0.05. RESULTS: For all three tests, root canals filled using epoxy resin-based sealer were not statistically significantly different compared with UDMA resin (P = 1 for Wf, P = 0.7 for MPSS and P = 0.12 for VRF), or different from both sound and prepared dentine (P > 0.05). There was also no significant difference between sound dentine and prepared dentine for both Wf (P = 0.92) and resistance to VRF (P = 1). CONCLUSIONS: Neither epoxy nor UDMA resins used as sealer cements enhanced fracture resistance of root dentine when placed within root canals of extracted teeth. PMID- 19134043 TI - Irrigant flow within a prepared root canal using various flow rates: a Computational Fluid Dynamics study. AB - AIM: To study using computer simulation the effect of irrigant flow rate on the flow pattern within a prepared root canal, during final irrigation with a syringe and needle. METHODOLOGY: Geometrical characteristics of a side-vented endodontic needle and clinically realistic flow rate values were obtained from previous and preliminary studies. A Computational Fluid Dynamics (CFD) model was created using FLUENT 6.2 software. Calculations were carried out for five selected flow rates (0.02-0.79 mL sec(-1)) and velocity and turbulence quantities along the domain were evaluated. RESULTS: Irrigant replacement was limited to 1-1.5 mm apical to the needle tip for all flow rates tested. Low-Reynolds number turbulent flow was detected near the needle outlet. Irrigant flow rate affected significantly the flow pattern within the root canal. CONCLUSIONS: Irrigation needles should be placed to within 1 mm from working length to ensure fluid exchange. Turbulent flow of irrigant leads to more efficient irrigant replacement. CFD represents a powerful tool for the study of irrigation. PMID- 19134044 TI - The management of mucosal fenestration: a report of two cases. AB - AIM: To report on the management of two patients with mucosal fenestration of root apices. SUMMARY: Two cases of mucosal fenestration of root apices were treated by a combination of nonsurgical root canal treatment and surgery. Root end resection was performed to bring the root apices within the alveolus before root-end filling and packing of the bony defects with demineralized freeze-dried bone allograft. The areas were then covered with connective tissue grafts taken from the hard palate before repositioning the mucogingival flap. At 1 year follow up, the mucosal fenestrations were completely healed by soft tissue graft coverage, and radiographs revealed bony healing. KEY LEARNING POINTS: * Connective tissue grafts can be used to manage mucosal fenestration. * Periradicular surgery and connective tissue grafts can be used in combination. PMID- 19134045 TI - Root canal morphology of the mesiobuccal root of maxillary first molars: a micro computed tomographic analysis. AB - AIM: To investigate ex vivo, the root canal morphology of the MB root of maxillary first molar teeth by means of micro-computed tomography. METHODOLOGY: Thirty extracted intact human maxillary first molar teeth were selected for micro tomographic analysis (SkyScan 1072, Aartselaar, Belgium) with a slice thickness of 38.0 mum. The following data regarding the MB root were analysed and recorded: number and type of root canals, prevalence of isthmuses, prevalence of intercanal connections, presence of accessory canals, presence of loops and number of apical foramina. RESULTS: The MB2 canal was present in 80% of specimens and was independent in 42% of these cases. When present, the MB2 canal merged with the MB1 canal in 58% of cases. Communications between the two canals were found in all specimens, with isthmuses in 71% of the cases. These communications and isthmuses were respectively in 42% and 54% of the cases in the coronal third, in 59% and 79% of the cases in the middle third and in 24% and 50% of the cases in the apical third. A single apical foramen was found in 37% of specimens, two apical foramina were present in 23% of the cases, with three or more separate apical foramina occurring in 40% of the specimens. CONCLUSIONS: The MB root canal anatomy was complex: a high incidence of MB2 root canals, isthmuses, accessory canals, apical delta and loops was found. PMID- 19134046 TI - Accidental injection with sodium hypochlorite: report of a case. AB - AIM: A case is reported in which sodium hypochlorite (NaOCl) was mistaken for anaesthetic solution and infiltrated into the buccal mucosa during routine root canal treatment. SUMMARY: A 1.5% sodium hypochlorite solution, kept in an anaesthetic cartridge, was inadvertently injected in the buccal mucosa of a 56 year-old female during routine root canal treatment. Soft tissue necrosis, labial ptosis and paraesthesia occurred shortly after the injection. Tissues healed with scarring and lip paraesthesia persisted for 3 years. KEY LEARNING POINTS: * NaOCl is highly irritant when introduced into oral tissues. * NaOCl solutions should not be kept in anaesthetic cartridges. * Accidents with NaOCl should be carefully assessed and when appropriate active hospital treatment should be sought. * Early recognition of NaOCl accidents may avert potentially more serious outcomes. PMID- 19134049 TI - Embryonic stem cells reduce liver fibrosis in CCl4-treated mice. AB - We transplanted undifferentiated embryonic stem (ES) cells into the spleens of carbon tetrachloride (CCl(4))-treated mice to determine their effects on liver fibrosis. Carbon tetrachloride at 0.5 ml/kg of body weight was injected intraperitoneally into C57BL/6 mice twice weekly for up to 20 weeks. Four weeks after the first injection, the mice were divided into two groups and those in group 1 received 1 x 10(5) ES cells genetically labelled with enhanced green fluorescent protein (GFP) in the spleens, while group 2 mice received 0.1 ml of phosphate-buffered saline. In group 1, GFP-immunopositive cells were retained and found in areas of fibrosis in the liver, and reduced liver fibrosis was observed as compared with group 2. Secondary transplantation of ES cells at 12 weeks after the initial transplantation enhanced the reduction in liver fibrosis. No teratoma formation or uncontrolled growth of ES cells in organs, including the liver and spleen, was observed in any of the mice. In the livers of group 1 mice, metalloproteinase 9-immunopositive cells derived from ES cells as well as those from the recipient were observed. These cells were also found to be immunopositive for the hepatoblast marker Delta-like (DlK-1), a member of the DlK 1 family of transmembrane proteins. These results suggest that ES-based cell therapy is potentially useful for liver fibrosis treatment and that reduction in CCl(4)-induced liver fibrosis by transplantation of ES cells may be related closely to the emergence of metalloproteinase-producing hepatoblast-like cells. PMID- 19134050 TI - Liver sinusoidal endothelial cells and acute non-oxidative hepatic injury induced by Pseudomonas aeruginosa pyocyanin. AB - The liver sinusoidal endothelial cell (LSEC) is damaged by many toxins, including oxidants and bacterial toxins. Any effect on LSECs of the Pseudomonas aeruginosa virulence factor, pyocyanin, may be relevant for systemic pseudomonal infections and liver transplantation. In this study, the effects of pyocyanin on in vivo rat livers and isolated LSECs were assessed using electron microscopy, immunohistochemistry and biochemistry. In particular, the effect on fenestrations, a crucial morphological aspect of LSECs was assessed. Pyocyanin treatment induced a dose-dependent reduction in fenestrations in isolated LSECs. In the intact liver, intraportal injection of pyocyanin (11.9 microM in blood) was associated with a reduction in endothelial porosity from 3.4 +/- 0.2% (n = 5) to 1.3 +/- 0.1% (n = 7) within 30 min. There were decreases in both diameter and frequency of fenestrations in the intact endothelium. There was also a decrease in endothelial thickness from 175.8 +/- 5.8 to 156.5 +/- 4.0 nm, an endothelial pathology finding previously unreported. Hepatocyte ultrastructure, liver function tests and immunohistochemical markers of oxidative stress (3 nitrotyrosine and malondialdehyde) were not affected. Pyocyanin induces significant ultrastructural changes in the LSEC in the absence of immunohistochemical evidence of oxidative stress or hepatocyte injury pointing to a novel mechanism for pyocyanin pathogenesis. PMID- 19134051 TI - An intravital microscopic study of the hepatic microcirculation in cirrhotic mice models: relationship between fibrosis and angiogenesis. AB - This intravital fluorescence microscopy (IVFM) study validates cirrhotic mice models and describes the different intrahepatic alterations and the role of angiogenesis in the liver during genesis of cirrhosis. Cirrhosis was induced by subcutaneous injection of carbon tetrachloride (CCl(4)) and by common bile duct ligation (CBDL) in mice. Diameters of sinusoids, portal venules (PV), central venules (CV) and shunts were measured at different time points by IVFM. Thereafter, liver samples were taken for sirius red, CD31, Ki67, vascular endothelial growth factor (VEGF) and alpha-smooth muscle actin (alpha-SMA) evaluation by immunohistochemistry (IHC). In parallel with fibrogenesis, hepatic microcirculation was markedly disturbed in CCl(4) and CBDL mice with a significant decrease in sinusoidal diameter compared to control mice. In CCl(4) mice, CV were enlarged, with marked sinusoidal-free spaces around CV. In contrast, PV were enlarged in CBDL mice and bile lakes were observed. In both mice models, intrahepatic shunts developed gradually after induction. During genesis of cirrhosis using CD31 IHC we observed a progressive increase in the number of blood vessels within the fibrotic septa area and a progressively increase in staining by Ki67, VEGF and alpha-SMA of endothelial cells, hepatocytes and hepatic stellate cells respectively. In vivo study of the hepatic microcirculation demonstrated a totally disturbed intrahepatic architecture, with narrowing of sinusoids in both cirrhotic mice models. The diameters of CV and PV increased and large shunts, bypassing the sinusoids, were seen after both CCl(4) and CBDL induction. Thus present study shows that there is angiogenesis in the liver during cirrhogenesis, and this is probably due partially to an increased production of VEGF. PMID- 19134052 TI - The influence of retrograde reperfusion on the ischaemia-/reperfusion injury after liver transplantation in the rat. AB - Dysfunction of the graft after liver transplantation caused by ischaemia /reperfusion (I/R) injury is a serious clinical problem. The aim of this study was to evaluate the influence of different kinds of reperfusion on I/R injury in a rat model. Arterialized orthoptic rat liver treatment was performed on male LEWIS-(RT(1))-rats. Three groups (n = 7) were formed. Group I: antegrade reperfusion with a 6-min delayed reperfusion via the hepatic artery. Group II: Antegrade reperfusion, simultaneously, via the portal vein and the hepatic artery. Group III: Retrograde reperfusion via the vena cava. Serum parameters were determined one, 24 and 48 h after operation. Furthermore, after 48 h, the liver was taken for histological assessment. After 48 h, rats of group III showed significantly lower aspartate amino transferase and alanine amino transferase serum levels compared with group I and group II rats. Forty-eight hours after transplantation, glutamate dehydrogenase serum level was significantly lower in group III than in group II. In histology, group III livers showed significantly less necrotic spots than group I and group II livers. Maximum size of the necrotic spots was significantly lower in group III than in group I. Also, significantly more necrotic spots were seen in the 'Rappaport's zone' 1 and 2 of group I than in group III. Our data suggested that the expression of I/R-injury correlates with the type of reperfusion. Furthermore, under standard conditions, this study was able to demonstrate that in a rat model, the retrograde reperfusion leads to a lower expression of I/R-injury than the antegrade reperfusion. PMID- 19134053 TI - Expression of galectins-1, -3 and -4 varies with strain and type of experimental colitis in mice. AB - Galectins are increasingly the focus of biomedical research. Although they are involved at different stages in inflammation, data on galectins in colitis remain scarce. The aim of this study was to determine and compare the expression of galectins in acute and chronic experimental colitis in mice. Immunohistochemistry for galectins-1, -3 and -4 was performed on colon tissue from C57BL/6 and BALB/c mice with acute dextran sodium sulphate colitis and from 129 Sv/Ev IL-10 knock out (IL-10(-/-)) mice. From these three mouse strains, we first detected major differences in galectin expression related to the genetic background in the control animals. With regard to inflammation, chronic colitis in IL-10(-/-) mice was associated with increased galectin-4 expression; in contrast with the two other models, no galectin-1 and -3 alterations were observed in IL-10(-/-) mice. Acute colitis in C57BL/6 and BALB/c mice showed increased galectin-3 expression in the lamina propria and the crypt epithelium, together with a decreased nuclear expression. These results suggest an involvement of galectins in the development and perpetuation of colonic inflammation and illustrate that the choice of the mouse strain for studying galectins might influence the outcome of the experiments. PMID- 19134054 TI - An in vivo rat model for early development of colorectal cancer metastasis to liver. AB - At diagnosis of colorectal cancer, approximately 25% of the patients have established colorectal liver metastasis. Optimal management of disseminated disease requires therapies targeting multiple stages in hepatic colorectal cancer metastasis development. To facilitate this, biologically accurate in vivo models are required. Early colonic cancer liver metastases development was studied using BDIX and Sprague-Dawley rat strains with human HT29 and rat DHDK12 colonic cancer cell lines. Different cancer cell-host combinations were used. Rat DHDK12 was previously chemically induced in the BDIX rat. Real-time intra-vital microscopy was employed to analyse the early development of liver metastases in four groups (n = 6 per group) (HT29-BDIX, DHDK12-BDIX, HT29-SD and DHDK12-SD). Data were compared using one-way anova with Bonferroni's multiple comparison test. The total number of tumour cells visualized, adherent cells within the hepatic sinusoids, extravasated tumour cells and migration rates were significantly higher in the DHDK12-BDIX combination. Maximum number of visualized cells and maximum migration rate were also significantly higher in this group. No significant differences were observed in these experimental parameters among the other three groups or in the haemodynamic parameters among all groups. In conclusion, cancer cell line-host selection has a significant effect on early colonic cancer liver metastasis development. PMID- 19134055 TI - Increased glial-derived neurotrophic factor in the small intestine of rats infected with the tapeworm, Hymenolepis diminuta. AB - The neurotrophin, glial-derived neurotrophic factor (GDNF), is essential for the development of the enteric nervous system (ENS) in both the embryo and neonate and may be important for maintenance and plasticity of ENS. The tapeworm, Hymenolepis diminuta, altered the number of cells containing GNDF in the host's jejunum and ileum. Numbers and locations of GDNF-containing cells were determined by applying monoclonal anti-GDNF antibody to intestinal segments collected from infected and uninfected age-matched rats during the initial 34 days post infection (dpi). Most cells staining positive for GDNF were present in the lamina propria of the jejunum and ileum from both infected and uninfected rats. The co localization of staining by the antibodies, anti-GDNF and anti-ED2 (a nuclear specific antibody for resident macrophages) indicated that at least 74% of the cells staining for GDNF were macrophages. Mast cells did not stain with the anti GDNF antibody. The increased number of GDNF+ cells in the infected rat intestine suggests that this neurotrophin may play a role in the neural and mucosal responses to lumenal tapeworm infection. PMID- 19134056 TI - Matrix metalloproteinase-9 contributes to intestinal tumourigenesis in the adenomatous polyposis coli multiple intestinal neoplasia mouse. AB - Matrix metalloproteinases (MMPs) are a family of 23 extracellular proteases that are best known for their collective ability to degrade all components of the extracellular matrix. We previously demonstrated that genetic ablation of MMP-7 reduced tumour multiplicity in multiple intestinal neoplasia (Min) mice possessing a genetic alteration in the adenomatous polyposis coli gene (APC). These mice, commonly referred to as APC-Min mice, are a frequently used model of early intestinal tumourigenesis. To examine further the role of MMPs in intestinal tumour development, we generated APC-Min mice genetically deficient in MMP-2, -9, -12 or -19. Genetic ablation of MMP-2, -12 or -19 did not affect multiplicity or size of intestinal tumours when crossed into the APC-Min system. However, MMP-9 deficient animals developed 40% fewer tumours than littermate controls, although tumour size distribution remained unaffected. Intestinal adenomas from MMP-9 deficient mice demonstrated a 50% decrease in proliferating cells compared with control tissues, with no difference in apoptosis. To determine the cellular origin of MMP-9 in these tumours, immunofluorescent co staining with markers for different leucocyte lineages was used to demonstrate that intratumoural MMP-9 is largely a product of neutrophils. These studies extend the potential targets for chemoprevention of intestinal adenomas to MMP-9 in addition to MMP-7 and exclude MMP-2,-12,-19 as attractive targets for intervention. PMID- 19134058 TI - 'Woundology'--an emerging clinical specialty. PMID- 19134057 TI - Acute colonic ischaemia in rats results in long-term structural changes without alterations of colonic sensitivity. AB - Colonic ischaemia and mast cells have been involved in the pathophysiology of the functional gastrointestinal disorder irritable bowel syndrome, although the cause effect relationships remain unknown. We assessed long-term histopathological and functional changes associated to an acute ischaemic episode (1 h) of the colon, followed by 8-week recovery, in rats. Functional colonic alterations [sensitivity during colorectal distension (CRD), compliance and propulsive motility] were assessed regularly during the recovery. Colonic histopathology (presence of inflammation, morphometric alterations and variations in neuronal density in the enteric nervous system) 8-week postischaemia was assessed. Following ischaemia, none of the functional parameters tested (motility, sensitivity and compliance) were affected. At necropsy, the colon presented an overall normal appearance with an increase in weight of the ischaemic area (mg/cm: 99 +/- 6; P < 0.05 vs. control: 81 +/- 4 or sham ischaemia: 81 +/- 3). Histopathological evaluations revealed the presence of a local infiltrate of mast cells in the area of ischaemia (nb of mast cells: 142 +/- 50; P < 0.05 vs. control, 31 +/- 14 or sham ischaemia: 40 +/- 16), without other significant alterations. Animals subjected to colonic ischaemia and treated 8 weeks later with the mast cell degranulator, compound 48/80, showed no changes in CRD-related pain responses. These studies show that acute colonic ischaemia is associated with the presence of a long-term local infiltration of mast cells, located within the serosa and muscle layers, despite the absence of functional changes, including colonic sensitivity. Considering the important pathophysiological functions of mast cells, the observed mast cell infiltration may be involved in ischaemia-induced functional changes yet to be characterized. PMID- 19134059 TI - Antibiotic-resistant gram-negative bacteria in deep tissue cultures. PMID- 19134060 TI - Wound care around the world. PMID- 19134061 TI - Hydrocolloid dressings in the management of acute wounds: a review of the literature. AB - A review of the literature suggests that the application of self-adhesive hydrocolloid dressings, most commonly associated with the treatment of ulcerative conditions such as pressure ulcers and leg ulcers, may also offer benefits in the management of acute wounds of all types, for example decreasing healing times of donor sites by about 40% compared with traditional treatments. Healing times of superficial traumatic injuries and surgical wounds are similarly enhanced but in the treatment of burns, the principal benefit appears to be a reduction in wound pain, an effect that has also been reported in virtually all other wound types. The impermeable nature of hydrocolloids provides a protective covering to the wound, permitting washing or showering while helping to prevent the spread of pathogenic microorganisms. There also appear to be significant cost-benefits associated with the use of hydrocolloids. In recent years, hydrocolloid dressings have been replaced by other products such as foams for the treatment of more heavily exuding wounds but for more lightly exuding wounds they still offer many practical advantages and as such will undoubtedly continue to meet an important need in wound management practice. PMID- 19134062 TI - Comparative analysis of global gene expression profiles between diabetic rat wounds treated with vacuum-assisted closure therapy, moist wound healing or gauze under suction. AB - How differential gene expression affects wound healing is not well understood. In this study, Zucker diabetic fatty (fa/fa) male inbred rats were used to investigate gene expression during wound healing in an impaired wound-healing model. Whole genome microarray surveys were used to gain insight into the biological pathways and healing processes in acute excisional wounds treated with vacuum-assisted closure (V.A.C.). Therapy, moist wound healing (MWH) or gauze under suction (GUS). Global gene expression analyses after 2 days of healing indicated major differences with respect to both number of genes showing fold changes and pathway regulation between the three different wound treatments. Statistical analysis of expression profiles indicated that 5072 genes showed a >1.6-fold change with V.A.C. Therapy compared with 3601 genes with MWH and 3952 genes with GUS. Pathways and related genes associated with the early phases of wound healing diverged between treatment groups. For example, pathways involving angiogenesis, cytoskeletal regulation and inflammation were associated with elevated gene expression following V.A.C. Therapy. This study is the first to assess wound healing by whole genome interrogation in a diabetic rat model treated with different healing modalities. PMID- 19134063 TI - Six-month mortality risks in long-term care residents with chronic ulcers. AB - Chronic ulcers are a common problem in long-term care. Residents with ongoing ulcers are often frail and at risk for mortality. This study evaluated the relationship between wound characteristics and other health predictors with 6 month mortality in nursing home residents. The subjects included were nursing home residents seen by the wound consult service from 1998 to 2007 with an ongoing chronic ulcer. This was a retrospective cohort study. Data were manually and electronically abstracted for each resident. Six-month mortality was collected as the primary outcome. Statistical comparisons were made using logistic regression with a final multivariant model. Four hundred and forty residents were seen with 411 records reviewed. Ulcer area was not associated with mortality; however, chronic ulcer number was associated with 6-month mortality with an odds ratio of 1.32 (95% CI 1.07-1.63). Other significant risk factors included heart failure, dementia, cancer, depression and blindness with all factors having an odds ratio greater than 1.75. Higher haemoglobin and venous insufficiency were protective of 6-month mortality. Ulcer number is an important predictor for 6-month mortality. The presence of multiple ulcers and comorbid health concerns may influence discussion of prognosis for healing and for potential end of life discussions. PMID- 19134064 TI - A comparative study of the in vitro permeation characteristic of sulphadiazine across synthetic membranes and eschar tissue. AB - Infections of burn wounds are the source of significant problems in burn patients. Early excision of eschar tissue is an ideal solution to avoid sepsis. When early excision is not feasible, the application of topical antimicrobial formulations may be used to control burn wound sepsis. An understanding of the barrier properties of eschar tissue is essential for optimal design of topical antimicrobial formulations. To date, little research has been conducted on the permeability of eschar. Silver sulphadiazine (SSD) is the most frequently used topical agent in burn management. In this study, the permeation of sulphadiazine from aqueous saturated solutions of SSD through human full-thickness burn eschar tissue was studied and compared with permeability through silicone and Carbosil as model membranes. The permeation of sulphadiazine through eschar tissue was significantly higher than that through silicone and Carbosil membranes (P < 0.05). Deconvolution of the data showed that the apparent sulphadiazine diffusion coefficient was much higher in eschar tissue and was comparable to transport through an aqueous protein gel. Further studies on a greater number of compounds are suggested to elucidate the utility of such membranes as predictive models of drug permeability through eschar tissue. PMID- 19134065 TI - Definition of efficiency in vacuum therapy--a randomised controlled trial comparing with V.A.C. Therapy. AB - Redon drains are still used to suction wounds for vacuum sealing. Vacuum-assisted closure (V.A.C.((R)); Kinetic Concepts Inc, San Antonio, TX) is a computer controlled therapy system for delivering topical negative pressure therapy. The efficiency of V.A.C. in the treatment of pressure ulcers was prospectively studied in a randomised controlled trial in which patients with pressure ulcers were randomly assigned to negative pressure wound therapy (NPWT) using either V.A.C. or Redon bottles. The target parameters were absolute and relative proportion of wound area consists of granulation tissue, fibrin and necrosis. Other outcome measures were the number of dressing changes and time invested using each system. The study was terminated after a post hoc analysis after inclusion of ten patients because of the significantly better results when using V.A.C., and the substantially larger care effort needed in the Redon group compared with the V.A.C. group. An increase in surface granulation tissue of 54% was observed in the V.A.C. group, and a reduction in the Redon group (P = 0.001). The Redon group showed an increase in fibrin tissue at the wound base of 21.8%, whereas in the V.A.C group, a 27% reduction was observed (P = 0.035). Necrosis was reduced in the V.A.C. group, but this difference did not reach significance. Redon bottles are not a good alternative for V.A.C. therapy for delivering NPWT. PMID- 19134066 TI - Tubercular infection in the sacrococcygeal pilonidal sinus--a case report. AB - We report a case of multiple pus discharging sinuses in the sacrococcygeal region in a 22-year-old man, who had a history of episodes of pain and pus discharge since 8 months. The patient underwent radiofrequency sinotomy using a Ellman radiofrequency device and was treated with antitubercular treatment, which resulted in complete resolution, and healing of the wound. PMID- 19134067 TI - The effects of education and training on clinical practice in wound healing. AB - This article considers the effects of two different types of educational programme on community nurse clinical practice in venous ulceration. One group of nurses (the experimental group) attended an educational programme designed to take account of training needs and learning styles. A second group of nurses (the control group) attended a standardised educational programme. A multiple-choice question examination and Objective Structured Clinical Examination were used to measure knowledge and skills. Kolb's Learning Styles Inventory was used to measure learning styles. Findings were that experimental nurses failed to show improved post-intervention clinical practice compared with the control group. PMID- 19134068 TI - Chitin membrane for wound dressing application--preparation, characterisation and toxicological evaluation. AB - Chitin, a unique biopolymer based on the N-acetyl-glucosamine monomer is envisioned to promote rapid dermal regeneration and accelerate wound healing. It has many useful and advantageous biological properties for its application as a wound dressing. Chitin membranes were prepared using lithium chloride/dimethylacetamide solvent system and evaluated for use as a wound dressing. Swelling behaviour, moisture vapour transmission rate, microbial impermeability and antimicrobial efficacy of the dressings was evaluated. The chitin dressing provided an effective barrier to microbial penetration and exerted a broad bacteriostatic action against Gram-positive and Gram-negative organisms. Gamma irradiation at 25 kGy was found suitable for sterilisation of the dressings. The thermal decomposition of unirradiated and irradiated chitin membranes was investigated. No significant change in the thermal behaviour because of irradiation at 25 kGy was observed. In vitro biodegradation of unirradiated and irradiated chitin membranes showed the susceptibility of the chitin dressing to lysozyme. Irritant effect of the chitin membrane dressings on skin was tested. Subcutaneous and scarification test in guinea pigs showed no signs of inflammation. This was further supported by the Finkelstein's test performed in rabbits. The chitin membranes were found to have optimal performance characteristics of a wound dressing and showed no toxicity or possible adverse reactions. The study shows the chitin dressings as useful adjunct in wound care. PMID- 19134069 TI - Changes in serum and exudate creatine phosphokinase concentrations as an indicator of deep tissue injury: a pilot study. AB - Deep tissue injury (DTI) is difficult to detect in the early phase. Creatine phosphokinase (CPK) as a muscle enzyme could represent a promising indicator of DTI. However, serum CPK levels reflect the systemic condition rather than the local wound environment. Wound exudates can be indicative of the local wound environment. This study aimed to investigate the usefulness of CPK levels in wound exudates as an indicator of DTI. Rats were divided into control, 6 hours 10 kg and 6 hours 20-kg loading groups. Serum samples were obtained before wounding, and at 8 and 12 hours, and 1, 2 and 3 days after wounding, while exudate samples were obtained on days 2 and 3. Serum CPK levels were markedly increased in the 10 kg and 20-kg groups at 8 and 12 hours after loading compared with the baseline value and control group, but decreased to the normal level on day 1. In both loading groups, exudate CPK levels were high on day 2 and decreased on day 3. Muscle necrosis was more severe in the 20-kg group than in the 10-kg group by histological examination. This is the first study to indicate the potential of CPK in wound exudates as an indicator of DTI. PMID- 19134070 TI - Estimation of genetic parameters: it takes three to tango. PMID- 19134071 TI - Reliable computing in estimation of variance components. AB - The purpose of this study is to present guidelines in selection of statistical and computing algorithms for variance components estimation when computing involves software packages. For this purpose two major methods are to be considered: residual maximal likelihood (REML) and Bayesian via Gibbs sampling. Expectation-Maximization (EM) REML is regarded as a very stable algorithm that is able to converge when covariance matrices are close to singular, however it is slow. However, convergence problems can occur with random regression models, especially if the starting values are much lower than those at convergence. Average Information (AI) REML is much faster for common problems but it relies on heuristics for convergence, and it may be very slow or even diverge for complex models. REML algorithms for general models become unstable with larger number of traits. REML by canonical transformation is stable in such cases but can support only a limited class of models. In general, REML algorithms are difficult to program. Bayesian methods via Gibbs sampling are much easier to program than REML, especially for complex models, and they can support much larger datasets; however, the termination criterion can be hard to determine, and the quality of estimates depends on a number of details. Computing speed varies with computing optimizations, with which some large data sets and complex models can be supported in a reasonable time; however, optimizations increase complexity of programming and restrict the types of models applicable. Several examples from past research are discussed to illustrate the fact that different problems required different methods. PMID- 19134072 TI - On identifiability of (co)variance components in animal models with competition effects. AB - There is an increased interest in estimating the (co)variance components of additive animal models with direct and competition effects (AMC). However, some attempts to estimate the dispersion parameters in different animal species faced problems of convergence or inaccurate estimates when pen effects entered the model. We argue that the problem relates to lack of identifiability of the (co)variance components in some AMC. The check for identifiability of the dispersion parameters in mixed models with linear (co)variance structure requires that all the eigenvalues of the restricted maximum likelyhood information matrix (I(theta)) be positive. We show, by way of simple numerical examples, that the singularity of I(theta) is due to confounding between fixed pen effects and the additive competition effects (SBVs). It is also observed that setting pen effects as random does not always remedy the collinearity with SBVs. An alternative AMC is presented in which the incidence matrix of the SBVs can be written as a function of the 'intensity of competition' (IC) among animals in the same pen. Examples are presented in which the ICs are related to time. The distribution of families of full and half sibs across pens also plays a role in the identifiability and asymptotic variances of the (co)variance components. PMID- 19134073 TI - Analysis of genotype by environment interaction for milk yield traits in first lactation of Simmental cattle. AB - The breeding goal for Simmental cattle is derived for intensively managed dairy farms. Its suitability for extensive farms was addressed by analysing possible genotype by environment interaction (G x E) between the management levels for first lactation milk yield traits. A first analysis was performed with the data collected from 300 000 purebred daughters of 278 second crop bulls born in Bavaria in 1993 and 1994. The farms were classified by herd-year-effect, using the sum of fat and protein yields into two levels of management, either with 33 or 10% quantiles, corresponding to approximately 100 000 cows and 30 000 cows, respectively. The comparison was based on 'daughter yield' deviations (DYD). Correlations between DYD of extensive and intensive environments were 0.90, 0.91 and 0.87 for milk, fat and protein yield (kg) for 33% quantiles, respectively. Corresponding correlations for 10% quantiles were 0.85, 0.83 and 0.77. Despite high correlations, 50 out of 149 sires showed significant differences between DYD in different environments. Bulls with higher DYD for milk yield on intensive farms were superior in all environments. For the second analysis extensive and intensive farms in northern and southern Bavaria were chosen at random. Approximately 20 000 cows in each management class were used for the estimation of genetic parameters. In both regions phenotypic and additive-genetic variances were higher in the intensively managed herds. Likewise heritabilities were higher for fat and protein yield, but not for milk where higher heritabilities were observed in 33% quantiles. Genetic correlations between extensive and intensive environments were 0.97 and above (33% quantiles). Ten per cent quantiles led to lower genetic correlations (0.90-0.95). Although no serious re-ranking effects of sires were evident, the scale effect and the differences in genetic parameters should be taken into consideration in practical breeding. PMID- 19134074 TI - Multiple trait genetic evaluation of ewe traits in Icelandic sheep. AB - The prolificacy of the ewes was measured as the number of lambs born per ewe mated (NLB) when the ewes were 1-4 years of age. The ewe productivity related to the same age interval was measured by special ewe production indices (EPI). The genetic parameters for these traits were estimated by a series of bivariate REML analyses using animal models. The material used for the genetic analysis contained records on 193,213 ewes. The heritability estimates for NLB were h(2) = 0.17, 0.13, 0.11, 0.10 for the four respective age classes. Corresponding estimates for EPI were h(2) = 0.16, 0.17, 0.17, 0.15. The genetic correlations among NLB at different ages ranged from 0.63 to 0.98 and among EPI from 0.82 to 0.99. The genetic correlations between NLB and EPI were generally low. The material used for estimating the breeding values by the MT-BLUP Animal Model consisted of 1.5 million individuals in the pedigree file. In total 815,782 ewes had records for the NLB and 763,491 ewes had production index (at least 1 year). The records were registered in the years 1990-2006. All possible missing patterns were present in the data. In the iteration process expected values for missing traits were generated and solutions were obtained on canonical transformed scale. The genetic evaluations were run independently for NLB and EPI for computational convenience given the correlations between these traits were negligible. PMID- 19134075 TI - Genetic correlations between ewe reproduction and carcass and meat quality traits in Merino sheep. AB - Genetic correlations between reproduction traits in ewes and carcass and meat quality traits in Merino rams were obtained using restricted maximum likelihood procedures. The carcass data were from 5870 Merino rams slaughtered at approximately 18 months of age that were the progeny of 543 sires from three research resource flocks over 7 years. The carcass traits included ultrasound scan fat and eye muscle depth (EMDUS) measured on live animals, dressing percentage and carcass tissue depth (at the GR site FATGR and C site FATC), eye muscle depth, width and area and the meat quality indicator traits of muscle final pH and colour (L*, a*, b*). The reproduction data consisted of 13 464 ewe joining records for number of lambs born and weaned and 9015 records for LS. The genetic correlations between reproduction and fat measurements were negative (range -0.06 +/- 0.12 to -0.37 +/- 0.12), with smaller correlations for live measurement than carcass traits. There were small favourable genetic correlations between reproduction traits and muscle depth in live rams (EMDUS, 0.10 +/- 0.12 to 0.20 +/- 0.12), although those with carcass muscle traits were close to zero. The reproduction traits were independent of meat colour L* (relative brightness), but tended to be favourably correlated with meat colour a* (relative redness, 0.12 +/- 0.17 to 0.19 +/- 0.16). There was a tendency for meat final pH to have small negative favourable genetic correlations with reproduction traits (0.05 +/- 0.11 to -0.17 +/- 0.12). This study indicates that there is no antagonism between reproduction traits and carcass and meat quality indicator traits, with scope for joint improvement of reproduction, carcass and meat quality traits in Merino sheep. PMID- 19134076 TI - Genetic variation and genetic trends in hip and elbow dysplasia in Swedish Rottweiler and Bernese Mountain Dog. AB - The aim of this study was to estimate genetic parameters and genetic trends for hip (HD) and elbow dysplasia (ED) in Swedish Rottweiler (RW) and Bernese Mountain Dog (BMD). Analyses were based on screening results of hip status for 14 693 RW and 8221 BMD and elbow status for 11 891 RW and 7963 BMD, as well as pedigree data for 16 614 RW and 9835 BMD, recorded by the Swedish Kennel Club. Components of (co)variance and breeding values were obtained with a mixed linear animal model. The model included the fixed effects of sex, birth month, age at screening and a combined random effect of clinic and year of examination. The need to include genetic groups for phantom parents in the model was evaluated by comparison of two different models: with and without genetic groups. Estimated heritabilities for HD and ED were between 0.34 and 0.42. The genetic correlation between the traits was weak and positive for RW (r(g) = 0.23 +/- 0.05) and not different from zero for BMD (r(g) = 0.06 +/- 0.06). F-statistics of the genetic group effects were not significant, implying that genetic groups do not need to be included in the model. Genetic trends indicated a genetic improvement in both traits. However, a faster genetic progress is expected if selection is based on predicted breeding values rather than phenotype. Based on the results, a statistical model for routine prediction of breeding values for HD and ED in Swedish dogs was suggested. PMID- 19134077 TI - Estimation of heritability for superficial digital flexor tendon injury by Gibbs sampling in the Thoroughbred racehorse. AB - The superficial digital flexor tendon (SDFT) injury causes considerable wastage of racing Thoroughbreds. In this study, the heritability of SDFT injury of the forelimbs was estimated in the Thoroughbred racehorses based on the Bayesian analysis with Gibbs sampling using threshold animal models. New horse patients of SDFT injury at the age of 2-5 years accommodated in the training centers in 2005 were used for the analyses. Based on clinical data of 8198 horses, heritability of SDFT was estimated to be the order of 0.17 (posterior mode) to 0.19 +/- 0.05 (posterior mean +/- posterior SD). PMID- 19134078 TI - Genetic diversity and structure of the West Balkan Pramenka sheep types as revealed by microsatellite and mitochondrial DNA analysis. AB - Several different phenotypes of the native Pramenka sheep have been developed in the Balkan region for different environmental and socio-cultural conditions. Animals from seven West Balkan Pramenka sheep types were analysed for 15 microsatellite markers and for mitochondrial DNA (mtDNA) and the results were used to assess genetic variation within and among the types and to infer the genetic population structure of the Pramenka sheep. Mean expected heterozygosity and allelic richness over the microsatellite loci and sheep types were 0.78 and 7.9, respectively. A Bayesian statistical method for estimating hidden genetic structure suggested that a core of the largest panmictic population was formed by Serbian, Kosovan, Bosnian, Montenegrin and Albanian types, while Croatian and Macedonian types comprised two other main populations, respectively. Mitochondrial DNA analysis revealed two mtDNA haplogroups in the Pramenka sheep, B and A, with a frequency of 93.7% and 6.3%, respectively. A total of 60 mtDNA haplotypes were found in 64 animals sequenced, and the mean nucleotide and haplotypic diversities over the types were 0.013 and 0.945, respectively. Molecular analysis suggests that the West Balkan Pramenka sheep types have their origins in two distinct maternal lineages of domestic sheep and different Pramenka phenotypes tend to form few panmictic populations. The Pramenka sheep represents a valuable resource of genetic diversity in sheep. PMID- 19134079 TI - Genetic variation at the alpha-1-fucosyltransferase (FUT1) gene in Asian wild boar and Chinese and Western commercial pig breeds. AB - Escherichia coli F18 bacteria producing enterotoxins and/or shigatoxin (ETEC/STEC) are main pathogens that cause oedema disease and postweaning diarrhoea in piglets, and alpha-1-fucosyltransferase (FUT1) gene has been identified as a candidate gene for controlling the expression of ETEC F18 receptor. The genetic variations at nucleotide position 307 in open reading frame of FUT1 gene in one wild boar breed and 20 western commercial and Chinese native pig breeds were investigated by polymerase chain reaction-restriction fragment length polymorphism. The results showed that the genetic polymorphisms of the FUT1 locus were only detected in western pig breeds and the Chinese Taihu (including Meishan pig, Fengjing pig and Erhualian pig), Huai and Lingao pig breeds; only Duroc and Pietrain possessed the resistant AA genotype, while the wild boar and other Chinese pig breeds only presented the susceptible genotype GG. The results indicated that Chinese native pig breeds lack genetic factors providing resistance to ETEC F18 bacteria. The resistant allele to ETEC F18 might originate from European wild boar. It was inferred that oedema and postweaning diarrhoea caused by ETEC F18 have close relationship with the growth rate, which can explain why on the contrary Chinese native pig breeds have stronger resistance to oedema and postweaning diarrhoea in piglets compared with western pig breeds. PMID- 19134080 TI - Evaluation of closed reduction and screw fixation in lag fashion of sacroiliac fracture-luxations. AB - OBJECTIVE: To evaluate outcome by radiographic assessment after closed reduction and percutaneous screw fixation in lag fashion of sacroiliac fracture-luxations in dogs. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=24) with sacroiliac fracture-luxations. METHODS: Medical records (1999-2006) and radiographs of 24 dogs (29 fracture-luxations) that had stabilization of sacroiliac fracture luxation by fluoroscopic-guided closed reduction and percutaneous screw fixation in lag fashion were reviewed. Signalment, body weight, number, and location of all concurrent injuries and implants used for repair were recorded. Radiographs were used to evaluate the accuracy of screw placement in the sacral body, screw depth/sacral width ratio, reduction of the sacroiliac joint, pelvic canal diameter, and hemipelvic canal width. Radiographic re-examination (range, 4 to >8 weeks postoperatively) was available for evaluation. RESULTS: Mean screw depth/sacral width ratio on immediate postoperative and re-examination radiographs was 64% and 61%, respectively. Mean percentage reduction of the sacroiliac joint on immediate postoperative and re-examination radiographs were 91% and 87%, respectively. Pelvic canal diameter ratio demonstrated successful restoration of the pelvic canal. Hemipelvic canal width ratio documented successful closed reduction repair independent of concurrent pelvic injuries. CONCLUSION: Successful repair of sacroiliac fracture-luxations, determined by radiographic assessment, can be achieved by fluoroscopic-guided closed reduction and percutaneous screw fixation in lag fashion. CLINICAL RELEVANCE: Fluoroscopic guided closed reduction and percutaneous screw fixation in lag fashion of sacroiliac fracture-luxations is a minimally invasive technique that restores and maintains pelvic canal dimensions and should be considered as an alternative to open reduction or nonsurgical management of sacroiliac fracture-luxations. PMID- 19134081 TI - Effect of head position on radiographic assessment of laryngeal tie-forward procedure in horses. AB - OBJECTIVE: To assess the effect of head position on relative position of the larynx and hyoid apparatus in horses with palatal dysfunction, and to define a standard position for radiographic assessment of laryngeal tie-forward. STUDY DESIGN: Prospective clinical study. ANIMALS: Adult horses (n=9) with palatal dysfunction. METHODS: Left lateral radiographs of the larynx were obtained pre and postoperatively for 3 different head positions (flexed=90 degrees ; neutral=100 degrees ; extended=115 degrees ). Distance between thyrohyoid bone and thyroid cartilage was measured. Data were analyzed to investigate differences between head positions, and to compare differences between pre- and postoperative measurements. RESULTS: Head position had a significant effect on relative position of the larynx and hyoid apparatus preoperatively. There was no significant difference in postoperative measurements of the 3 head positions. A significant difference between pre- and postoperative measurements was found with the head in the neutral or extended position whereas there were no significant differences between pre- and postoperative measurements using the flexed position. CONCLUSIONS: Head position affects the relationship between the hyoid apparatus and larynx preoperatively, with greatest distance occurring when the head is extended. In a flexed position, it is not possible to ascertain whether laryngeal position has changed postoperatively. CLINICAL RELEVANCE: A standardized head position is necessary when assessing the laryngeal tie-forward procedure radiographically. An extended head position is most useful for this assessment. PMID- 19134082 TI - Osteochondral fragmentation in the synovial pad of the fetlock in Warmblood horses. AB - OBJECTIVES: To determine clinical and arthroscopic characteristics associated with fragments in the synovial pad of the fetlock and to characterize their morphology. STUDY DESIGN: Retrospective study. ANIMALS: Warmblood horses (n=104) with fragment(s) in the synovial pad. METHODS: Signalment and results of radiographic and clinical examination were collected before surgery. After arthroscopic fragment removal and joint evaluation for synovial and/or cartilage abnormalities, fragments were measured, and evaluated by histopathology. RESULTS: Synovial pad fragments (n=142) were removed from 127 fetlocks. Two older horses had lameness. During arthroscopy, abnormalities were observed in 40 joints (31.5%) and multivariate logistic regression analysis showed a significant correlation between the observed arthroscopic abnormalities and the presence of large fragments (P=.016). Fragments were osteochondral bodies completely surrounded by fibrous tissue. At the edges of the hyaline cartilage cap an underlying fibrous structure was obvious in the extracellular matrix giving it a reactive pattern. CONCLUSIONS: Although the impact on lameness was minimal, there was a significant correlation between arthroscopic abnormalities and presence of large synovial pad fragments. On histopathology, these osteochondral fragments are embedded in fibrous tissue and show a reactive pattern. They are not a manifestation of any well-described joint pathology. CLINICAL RELEVANCE: Large synovial pad fragments in Warmblood horses can be associated with synovial and cartilage abnormalities, but further studies are warranted to determine their origin and clinical importance. PMID- 19134083 TI - Surgical approach for removal of nonarticular base sesamoid fragments of the proximal sesamoid bones in horses. AB - OBJECTIVES: To describe a surgical approach for the removal of nonarticular base sesamoid fragments in performance horses and to report the outcome. STUDY DESIGN: Retrospective study. ANIMALS: Horses (n=11), 7 months to 10 years of age, with lameness because of nonarticular base sesamoid fragments. METHODS: Lameness was localized to the metacarpophalangeal/metatarsophalangeal region by clinical examination or response to diagnostic local anesthesia. Radiographs confirmed the diagnosis of a nonarticular base sesamoid fragment. Surgical removal was performed with an incision over the base of the affected sesamoid through the digital flexor tendon sheath. After identifying the fragment with the aid of needles, a small vertical incision was made in the straight distal sesamoidean ligament (SDSL) and the fragment was freed from its attachments and removed. Six months convalescence and rehabilitation was prescribed for all horses. RESULTS: Eleven horses had 16 nonarticular fragments of the base of the proximal sesamoid bones. Eleven (69%) fragments occurred in the forelimbs with the right front (82%) and medial sesamoid (73%) more commonly affected. Horses were treated by surgical fragment removal. In horses with follow-up, 9 of 10 returned to their intended use. CONCLUSIONS: Surgical removal of nonarticular base sesamoid fragments can be accomplished through a palmar/plantar approach through the digital flexor tendon sheath and SDSL. This "keyhole" approach minimizes damage to the distal sesamoidean ligament attachments to the sesamoid base and allows some horses to return to their intended use. CLINICAL RELEVANCE: Surgical removal of nonarticular base sesamoid fragments should be considered in horses with performance-limiting lameness as a result of the fragment. PMID- 19134084 TI - Laparoscopic cholecystectomy for management of uncomplicated gall bladder mucocele in six dogs. AB - OBJECTIVES: To describe a technique for, and outcome after, laparoscopic cholecystectomy (LC) for management of uncomplicated gall bladder mucocele (GBM) in dogs. STUDY DESIGN: Case series. ANIMALS: Dogs (n=6) with uncomplicated GBM. METHODS: Dogs with ultrasonographic evidence of GBM but without imaging or laboratory signs of gall bladder rupture, peritonitis, or extra-hepatic biliary tract rupture that had LC were included. A 4 portal technique was used. A fan retractor was used to retract the gall bladder to allow dissection around the cystic duct with 5 or 10 mm right-angle dissecting forceps. The cystic duct was ligated using extracorporeally tied ligatures supplemented sometimes with hemostatic clips. A harmonic scalpel was used to dissect the gall bladder from its fossa. The gall bladder was placed into a specimen retrieval bag and after bile aspiration the bag was withdrawn through the 11 mm portal incision. RESULTS: Five dogs had mild intermittent clinical signs including vomiting, inappetence, and lethargy. All dogs had successful LC without conversion to an open approach. All dogs with clinical signs had improvement or resolution of signs postoperatively. No important perioperative complications occurred and all dogs were alive at a median of 8 months postoperatively (range, 3-14 months). CONCLUSIONS: LC can be accomplished safely and effectively in dogs with uncomplicated GBM. CLINICAL RELEVANCE: A minimally invasive approach for cholecystectomy can be used for the treatment of GBM in dogs. PMID- 19134085 TI - Magnetic resonance imaging findings in dogs with confirmed shoulder pathology. AB - OBJECTIVE: To evaluate the diagnostic potential of magnetic resonance imaging (MRI) compared with a reference standard, arthroscopic and/or open surgery, in dogs with soft tissue shoulder pathology. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=21). METHODS: Magnetic resonance (MR) images were retrospectively evaluated in 21 dogs that had surgically identified soft tissue shoulder pathology. The musculotendinous units of the biceps, infraspinatus, teres minor, supraspinatus, subscapularis, and the medial and lateral glenohumeral ligaments (MGHL and LGHL) were graded as either normal or abnormal. Abnormal structures were further classified as being either inflamed, partially torn, or fully torn. Impingement of the biceps tendon was also evaluated. Results were reported in terms of agreement and concordance between MRI findings and surgical findings. Agreement was defined as the percentage of times MRI findings concurred with surgical findings with respect to a structure being either normal or abnormal. Concordance was defined as the percentage of times MRI concurred with the exact surgically assessed pathology when abnormality was identified. RESULTS: The findings were biceps tendon: 90% agreement with 100% concordance; subscapularis: 95% agreement with 62% concordance; MGHL: 84% agreement with 83% concordance; LGHL: 88% agreement with 100% concordance; infraspinatus: 100% both agreement and concordance; biceps tendon impingement: 90% agreement with 100% concordance. CONCLUSIONS: Soft tissue abnormalities of the canine shoulder were readily identified on preoperative MR images. CLINICAL RELEVANCE: MRI shows great potential as a diagnostic tool in the evaluation of canine shoulder disease. PMID- 19134086 TI - Clinical applications of demineralized bone matrix: a retrospective and case matched study of seventy-five dogs. AB - OBJECTIVES: To evaluate the outcome in dogs treated with demineralized bone matrix (DBM) as an adjunct to orthopedic procedures. STUDY DESIGN: Retrospective and case-match study. ANIMALS: Dogs (n=75). METHODS: Medical records (1999-2006) and radiographs of dogs that had orthopedic procedures (comminuted fractures, tibial plateau leveling osteotomy [TPLO] where correction for tibial rotation created an osteotomy gap, arthrodeses, open corrective osteotomies) where DBM was used were reviewed for signalment, quantity of DBM implanted, duration of exercise restriction, radiographic healing, and complications. Dogs that had TPLO and correction of tibial torsion (n=15), or arthrodesis (n=16) were compared with case-matched controls. Data were analyzed using Kruskal-Wallis test, ANOVA, Tukey's HSD test, and logistic regression analysis. RESULTS: Mean (+/-SD) healing time for orthopedic surgeries with DBM augmentation were 15+/-6.97 (weeks) and complication rate was 19% (14 dogs). Dogs with a TPLO gap filled with DBM were allowed to return to normal exercise 2 weeks earlier than dogs with a well apposed TPLO site. Radiographic healing, duration of exercise restriction, and timing of destabilization were similar in dogs undergoing carpal and tarsal arthrodesis whether they received DBM, autogenous graft, or both. CONCLUSIONS: DBM can be used to treat uncomplicated bone defects associated with comminuted fracture repairs, open osteotomies, and arthrodeses in dogs. Under these circumstances, clinicians might expect similar clinical outcomes without the possibility of side effects associated with the harvest of autogenous cancellous bone. CLINICAL RELEVANCE: DBM is safe for use in dogs. PMID- 19134087 TI - Acceleration of second and fourth metatarsal fracture healing with recombinant human bone morphogenetic protein-2/calcium phosphate cement in horses. AB - OBJECTIVE: To compare the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2)/calcium phosphate (CP) to autogenous cancellous bone graft (CBG) and to no treatment on bone healing, in surgically induced osteotomies and ostectomies of the accessory metatarsal bones in an equine model. STUDY DESIGN: Experimental. ANIMALS: Adult horses (n=9). METHODS: Segmental ostectomies of the second metatarsal bone (MT2) and osteotomies of the fourth metatarsal bone (MT4) were performed bilaterally in 9 horses. There were a total of 35 defects (1 MT4 was previously fractured) created and supplemented randomly either with no treatment (untreated control), rhBMP-2/CP cement, or matrix (CPC or CPM), or CBG. Radiography was performed every 2 weeks until study endpoint at 12 weeks. After euthanasia, bone healing was evaluated using radiography, mechanical testing, and histology. Data were analyzed with ANOVA followed by the Duncan's Multiple Range Test or nonparametric analyses. RESULTS: At 12 weeks, radiographic scores for union were significantly greater for the rhBMP-2 (P<.0001) and CBG (P=.004) groups compared with the untreated control group, for both MT2 ostectomies and MT4 osteotomies. The rhBMP-2 treated MT2 had greater maximum torque to failure in torsion than CBG and control limbs at 12 weeks (P=.011). Histologic analysis demonstrated increased bone formation and more mature bone at the ostectomy site for MT2 in the rhBMP-2 and CBG groups compared with the untreated control group. CONCLUSION: Injection of rhBMP-2/CP into surgically induced ostectomies and osteotomies of the accessory metatarsal bones might accelerate early bone healing in the horse. CLINICAL RELEVANCE: RhBMP-2/CP may be as effective if not superior to CBG as an adjuvant treatment to accelerate healing of bone defects. PMID- 19134088 TI - Liver size, bodyweight, and tolerance to acute complete occlusion of congenital extrahepatic portosystemic shunts in dogs. AB - OBJECTIVE: To investigate the relationship between preoperative liver size, bodyweight, and tolerance to shunt occlusion in dogs with congenital extrahepatic portosystemic shunt(s) (CPSS). STUDY DESIGN: Longitudinal cohort study. ANIMALS: Dogs with CPSS (n=35). METHODS: Ultrasonography was used to measure preoperative maximum transverse dimension of the liver (TS) of each dog. Intraoperative portal pressures were measured, before and after CPSS occlusion, via a jejunal vein catheter. Tolerance to shunt occlusion was judged on gross visceral observations, and on changes in portal pressure, central venous and mean arterial pressures. RESULTS: TS was significantly related to bodyweight (P<.05). Mean ratios for TS/bodyweight were calculated for dogs tolerant and intolerant of acute complete shunt occlusion. Dogs tolerant to occlusion had significantly higher TS/bodyweight ratios than dogs intolerant to occlusion (P=.025). Dogs with a TS/bodyweight ratio of >7 were more likely to tolerate CPSS occlusion than dogs with a TS/bodyweight ratio of <5 (P=.036). A model was generated to predict portal pressure rise after shunt occlusion, based on liver dimensions and bodyweight (R=0.668). Intestinal oxygenation did not correlate significantly with tolerance to CPSS occlusion (P=.29). CONCLUSION: In dogs with CPSS, liver size (relative to bodyweight) is significantly greater (P=.025) in dogs that are tolerant of full ligation than intolerant of occlusion. CLINICAL RELEVANCE: Preoperative measurement of bodyweight and liver size help indicate the likelihood of tolerance to acute complete occlusion of CPSS in dogs. PMID- 19134089 TI - Mechanical comparison of 3.5 mm broad dynamic compression plate, broad limited contact dynamic compression plate, and narrow locking compression plate systems using interfragmentary gap models. AB - OBJECTIVES: To compare (1) pullout properties between 3.5 mm cortical and locking screws, and (2) mechanical properties and gap displacements between the 3.5 mm broad limited-contact dynamic compression plate (LC-DCP), broad dynamic compression plate (DCP), and narrow locking compression plate (LCP), during axial loading of plate-stabilized diaphyseal fragments with an interfragmentary gap. STUDY DESIGN: In vitro mechanical testing of implanted polyurethane foam (PUF) hollow cylinders that simulated compact or osteopenic diaphyseal bone. SAMPLE POPULATION: (1) Five cortical and locking screws and (2) 4 PUF-plate constructs for each plate type; using high- and low-density (0.8 and 0.32 g/cm(3)) cylinders. METHODS: (1) Screws were completely extracted at 5 mm/min. (2) Plated constructs were axially compressed at 300 N/s for 10 cycles from 5 to 355 N to determine gap displacement during physiologic loading, followed by single cycle increasing load to failure. RESULTS: Pullout properties were not different between screw types. All plate constructs had yield loads over 3 times trotting loads. Gap closure occurred with LC-DCP and DCP constructs, but not LCP constructs. LCP construct properties were most similar to LC-DCP and DCP construct properties in the low-density model. CONCLUSION: All plate systems sustained physiologic limb loads. Only LCP constructs maintained some gap integrity, although LC-DCP and DCP screws were placed in neutral position. CLINICAL RELEVANCE: The LCP system is more likely than LC-DCP and DCP systems, with neutrally positioned screws, to maintain a planned interfragmentary gap, although gap strains range from 0% to 15% across the 2 mm gap during a trot load. PMID- 19134090 TI - Determination of pelvic limb alignment in the large-breed dog: a cadaveric radiographic study in the frontal plane. AB - OBJECTIVES: To report a method for radiographic determination of the mechanical axis of the canine pelvic limb and its relationship to the joints and bone axes. To report reference ranges for the relationships between the axes of the pelvic limb and for joint position relative to the pelvic limb axis. STUDY DESIGN: Cadaveric radiographic anatomic study. ANIMALS: Pelvic limbs (n=101) from normal midsized to large breed dogs; tibiae (n=105) from dogs with cranial cruciate ligament disease (previous study). METHODS: Extended full-limb radiography was performed and images analyzed to determine: mechanical joint reference angles (femur, tibia), pelvic limb axis, tibiofemoral and metatarsotibial angle, mechanical axis-femur/metatarsus angle, and mechanical axis deviation (MAD) of the stifle/tarsus. RESULTS: Mean mechanical angles were: lateral proximal femoral (103.7 degrees +/-5.4 degrees ), lateral distal femoral (98.6 degrees +/-2.5 degrees ), medial proximal tibial (92.2 degrees +/-1.8 degrees ), medial distal tibial (95.9 degrees +/-2.2 degrees ), tibiofemoral (9.1 degrees varus +/-2.8 degrees ), metatarsotibial (0.6 degrees valgus +/-2.1 degrees ). Mean mechanical axis-femur and-metatarsus angles were 5.6 degrees (+/-1.7 degrees ) and 2.9 degrees (+/-1.5 degrees ), respectively. Mean MAD of the stifle and tarsus were 3.6% (+/-1.1%) and 1.2% (+/-0.6%), respectively. Tibial angles were not different between dogs with and without cranial cruciate ligament disease. CONCLUSIONS: Mechanical axes of the canine pelvic limb and their relationship to the joints can be determined by full-limb radiography. CLINICAL RELEVANCE: Techniques and reference ranges may be useful for diagnosis, surgical planning, and postoperative assessment of pelvic limb deformities. PMID- 19134091 TI - Survival, complications, and analysis of risk factors after renal transplantation in cats. AB - OBJECTIVE: To report survival, complications, and analyze risk factors for survival after renal transplantation (RTr) and cyclosporine-A based immunosuppression in cats. STUDY DESIGN: Historical cohort. ANIMALS: Cats (n=60). METHODS: Data were obtained from medical records of cats that had RTr. Influence of various perioperative factors on survival and complications was evaluated. Occurrence of postoperative hypertension (HT), seizures, infection, acute allograft rejection (AR), congestive heart failure (CHF), and delayed graft function (DGF) was evaluated. RESULTS: Survival to discharge after RTr was 77.5%. Estimated median overall survival time was 613 days; 6 month and 3 year overall survival proportions were 65% and 40%, respectively. Age, weight, and blood pressure influenced overall survival. Increased preoperative creatinine concentration, blood urea nitrogen, postoperative creatinine concentration, left ventricular wall thickness, and reduced creatinine reduction ratio influenced survival until discharge. HT was identified in 9/30 (30%) cats; however, no risk factors were identified, nor was HT related to seizures. AR was identified in 8/62 (13%) grafts. Infection, predominantly bacterial, developed in 22/60 (37%) cats. CHF occurred in 7/60 (12%) cats before discharge. Cats experiencing CHF were younger, had an increased incidence of heart murmurs, and poor initial graft function. DGF was identified in 5 cats and seizures in 2 cats. CONCLUSIONS: RTr affords cats with CRF long survival times. Older cats and cats with severe azotemia, HT, and cardiovascular disease may have increased mortality after RTr. Complications after RTr were common. CLINICAL RELEVANCE: Clinicians should be aware of these risk factors when recommending feline RTr. PMID- 19134092 TI - Efficacy of en bloc ligation of the thoracic duct: descriptive study in 14 dogs. AB - OBJECTIVE: To assess whether en bloc ligation of all caudal mediastinal tissue between the aorta and thoracic vertebrae will occlude all branches of the thoracic duct. STUDY DESIGN: Descriptive study. ANIMALS: Canine cadavers (n=15). METHODS: The cisterna chyli was approached through a right paralumbar abdominal incision and cannulated. A right 10th intercostal thoracotomy was performed and the tissue of the caudal mediastinum dorsal to the aorta and ventral to the thoracic vertebrae was ligated. Radio-opaque contrast material was injected into the cisterna chyli during fluoroscopic imaging of the thorax and cranial abdomen. Still images were obtained before and after contrast injection and a subtraction image produced. Gray values were measured from these images, cranial and caudal to the ligature site. RESULTS: Cannulation of the cisterna chyli in 1 cadaver was impossible and it was, therefore, excluded. In 13 (93%) cadavers contrast material did not pass cranial to the ligature site. In 1 cadaver, an intact thoracic duct was identified on lymphangiography outside the en bloc ligation. CONCLUSION: En bloc ligation of the caudal mediastinal tissue dorsal to the aorta was successful in preventing opacification with contrast material of all branches of the thoracic duct in 93% of cadavers. CLINICAL RELEVANCE: En bloc ligation is successful in 93% of cadavers and reduces the operative time compared with techniques requiring some type of mesenteric lymphangiography to aid identification of the thoracic duct. PMID- 19134093 TI - Comparison of holding power of three different pin designs for external skeletal fixation in avian bone: a study in common buzzard (Buteo buteo). AB - OBJECTIVE: (1) To evaluate resistance to axial extraction of 3 pin designs in avian humerus and tibiotarsus; (2) to assess the effect of pin location within the bone on holding power; and (3) to assess the influence of thread pitch on holding power. STUDY DESIGN: Resistance of pins to axial extraction was measured immediately after insertion. ANIMALS: Adult common buzzards (Buteo buteo; n=9). METHODS: Different pin designs (1 smooth; 2 threaded pins, differing in pitch) were inserted into the proximal and distal metaphysis and the proximal, middle, and distal diaphysis of the humerus and tibiotarsus. Maximum force required for axial extraction of pins was recorded. RESULTS: Smooth pins had the lowest extraction force (P<.05). Pins inserted into the diaphysis (proximal, middle and distal) of the humerus and the distal metaphysis of the tibiotarsus had a greater pullout strength than pins in other locations. Pins with a smaller pitch inserted into the proximal diaphysis and distal metaphysis of the humerus, and the proximal metaphysis of the tibiotarsus had significantly greater holding power than pins with a larger pitch (P<.05). CONCLUSIONS: Pins inserted into the diaphysis of humerus and the distal metaphysis of the tibiotarsus are better at resisting extraction. Pins with a smaller pitch possess greater holding power than pins with a larger pitch in avian humerus and tibiotarsus. CLINICAL RELEVANCE: Consideration should be given to pin location and thread pitch, when choosing external skeletal fixation to repair an avian humeral or tibiotarsal fracture. PMID- 19134094 TI - Use of concordance correlation coefficient in objective and subjective assessment of limb function. PMID- 19134095 TI - Evidence-based practice. PMID- 19134098 TI - Surgical and conservative management of bilateral dynamic laryngeal collapse associated with poll flexion in harness race horses. AB - OBJECTIVE: To test the hypothesis that in bilateral dynamic laryngeal collapse associated with poll flexion, vocal fold collapse (VFC) is the initial abnormal event that induces further laryngeal collapse, and that racing performance would therefore be substantially improved after bilateral ventriculocordectomy in affected individuals. STUDY DESIGN: Retrospective study. ANIMALS: Twenty-six horses. METHODS: Medical records (1998-2006) of harness racehorses admitted for high-speed treadmill videoendoscopy (HSTV) that had bilateral dynamic laryngeal collapse associated with poll flexion were reviewed. Race records, owner interviews, and follow-up HSTV were used to evaluate outcome after either surgical treatment including bilateral ventriculocordectomy or conservative management. RESULTS: Bilateral dynamic laryngeal collapse, defined as bilateral VFC with concurrent arytenoid cartilage collapse (ACC), was identified in 26 horses. Norwegian Coldblooded Trotters (NCT) were overrepresented. Sixteen horses had surgical treatment and 10 were treated conservatively. Return to racing and racing performance was not improved within or between groups after the treatment. On follow-up HSTV of 6 surgically treated horses, there was no residual soft tissue collapse in the ventral portion of the rima glottidis; however, ACC and other abnormalities were still evident. CONCLUSION: Bilateral ventriculocordectomy resolved VFC, but failed to stabilize the arytenoid cartilages or to significantly improve racing performance. CLINICAL RELEVANCE: Bilateral dynamic laryngeal collapse associated with poll flexion is a serious performance-limiting upper respiratory tract disorder that is overrepresented in NCT racehorses. Our results suggest that VFC is not the initiating event in this complex obstructive airway disorder for which there is currently no consistently effective treatment. PMID- 19134099 TI - Locking compression plates for pancarpal arthrodesis in a Thoroughbred filly. AB - OBJECTIVE: To report treatment of a comminuted ulnar carpal (UC) bone fracture associated with carpal instability by pancarpal arthrodesis using 2 locking compression plates (LCP). STUDY DESIGN: Case report. ANIMALS: A 2-year-old Thoroughbred filly. METHODS: An UC fracture and luxation of the proximal row of carpal bones was diagnosed radiographically. Pancarpal arthrodesis was performed with 2 LCP positioned dorsolaterally and dorsomedially and centered over the carpus through a single skin incision. The filly was maintained in a full limb cast for 15 days, followed by a tube cast for 14 days, and subsequently a full limb bandage with caudal splint for 21 days. RESULTS: Two LCP provided appropriate carpal stability resulting in a pasture sound horse 6 months after surgery. The filly was discharged from the hospital 63 days after surgery, walking well with only a slight mechanical lameness. CONCLUSION: Use of 2 LCP applied on the dorsomedial and dorsolateral aspect of the carpus can provide carpal stability for pancarpal arthrodesis. CLINICAL RELEVANCE: Excellent stability of the carpal joints can be achieved with 2 LCP. PMID- 19134100 TI - Effect of porcine small intestinal submucosa on acute full-thickness wounds in dogs. AB - OBJECTIVE: To evaluate the effects of porcine small intestinal submucosa (PSIS) on the healing of full-thickness wounds in dogs, specifically the appearance of granulation tissue, percent epithelialization and contraction, histologic variables of inflammation and repair, and aerobic culture results. STUDY DESIGN: Prospective, controlled, experimental study. ANIMALS: Purpose-bred, female dogs (n=10). METHODS: Wounds were created bilaterally on the trunk; 1 side as a control and 1 treated with PSIS. First appearance of granulation tissue was recorded. Total wound area, open wound area, and epithelialized area were measured at 21 time points-wound contraction and percent epithelialization were calculated. Aerobic cultures were taken at 4 time points and wound biopsies at 8. Histologic features were graded into an Acute Inflammation Score and Repair Score. RESULTS: There was no difference in first appearance of granulation tissue between PSIS-treated and control wounds. Wound contraction was significantly faster in control wounds as was percent epithelialization after day 21. Histologic Acute Inflammation Scores were significantly higher in PSIS-treated wounds compared with control wounds on days 2 and 6. There were no differences in Histologic Repair Scores between PSIS-treated and control wounds or in aerobic culture results. CONCLUSION: Wounds treated with PSIS contract more slowly, epithelialize less, and have more pronounced acute inflammation after implantation than control wounds. CLINICAL RELEVANCE: Acute, full-thickness wounds in dogs do not benefit from treatment with PSIS. PMID- 19134101 TI - Validation of a technique to assess radial torsion in the presence of procurvatum and valgus deformity using computed tomography: a cadaveric study. AB - OBJECTIVE: To validate a technique to assess radial torsion with and without procurvatum and valgus deformity using computed tomography (CT). STUDY DESIGN: Anatomic study. ANIMALS: Radii (n=3) from normal dogs. METHODS: Three cadaveric radii were scanned using CT (3 mm slices). Transverse image sections were assessed at the distal and proximal physeal scar parallel to the joint surface in both sagittal and dorsal planes. Anatomic landmarks of the radius were identified and torsion was measured. A hinged Ilizarov circular external skeletal fixator was constructed to create known procurvatum, valgus, and torsional deformities. RESULTS: Between observers results were highly correlated for calculation of torsion (r=0.972, r(2)=0.9448, P<.00001). Mean baseline torsion of the radius in the construct was 4.88 degrees . At 20 degrees procurvatum and valgus, mean calculated torsion was 5.07 degrees (confidence interval [CI]=4.6-5.6 degrees ). At 20 degrees procurvatum and valgus with 22.5 degrees -induced torsion, mean measured torsion was 25.4 degrees (expected=27.38 degrees ; CI=22.7-28.1 degrees ). At 40 degrees procurvatum and valgus and 45 degrees -induced torsion, mean measured torsion was 51.1 degrees (expected=49.88 degrees ; CI=-49.2 to 53.0 degrees ). CONCLUSION: Assessment of radial torsion by CT is reproducible and accurate in the presence of procurvatum and valgus antebrachial angular limb deformities in the dog. CLINICAL RELEVANCE: Despite complex presurgical planning, current measurements only allow for measurement of valgus and procurvatum deformities in the sagittal and dorsal planes, this method allows for more precise measurements of radial torsion in the transverse plane. PMID- 19134102 TI - Cervical spinal locking plate in combination with cortical ring allograft for a one level fusion in dogs with cervical spondylotic myelopathy. AB - OBJECTIVE: To evaluate use of a surgical technique commonly used in humans for treatment of cervical spondylotic myelopathy (CSM) in dogs. DESIGN: Prospective case series. ANIMALS: Dogs with CSM (n=10). METHODS: Dogs weighing >30 kg that had CSM at 1 vertebral articulation were eligible for inclusion. Dogs had vertebral column distraction/fusion performed using a cortical ring allograft, cancellous autograft, and a spinal locking plate. Dogs were evaluated temporally by repeat neurological examinations and by client perception of postsurgical outcome, determined by telephone interview. RESULTS: Nine dogs survived the immediate postoperative period. Seven of 8 dogs had moderate to complete improvement without recurrence (mean follow-up, 2.48 years). The most common postsurgical complications were screw loosening (n=4) and plate shifting (2), neither of which required surgical revision. One dog had pseudoarthrosis that may have negatively impacted outcome. CONCLUSION: Treatment of single level CSM in dogs with ring allograft and a spinal locking plate system may lead to successful outcomes. The major problems encountered with included cost of the implants and adjusting the system designed for humans to fit the vertebral column of a dog. CLINICAL RELEVANCE: For dogs with CSM at a single level, the use of a spinal locking plate in combination with a cortical ring allograft can be an effective surgical treatment. Costs of the implants as well as anatomic differences in dogs make this type of surgery less appealing. PMID- 19134103 TI - Effect of preoperative erythromycin or dexamethasone/vitamin C on postoperative abomasal emptying rate in dairy cows undergoing surgical correction of abomasal volvulus. AB - OBJECTIVE: To determine the effects of preoperative erythromycin or combined dexamethasone/vitamin C treatment on postoperative abomasal emptying rate in cows undergoing surgical correction of abomasal volvulus (AV). STUDY DESIGN: Prospective, controlled, clinical study using a convenience sample. ANIMALS: Lactating Holstein-Friesian cows (n=45) with AV were alternately assigned to 3 groups (n=15): group C: untreated (control); group E: erythromycin (10 mg/kg intramuscularly [IM]); group D: dexamethasone (0.02 mg/kg intravenously [IV]) and vitamin C (10 mg/kg IV). METHODS: Drugs were administered 1 hour before surgical correction of AV. D-xylose solution (50%, 0.5 g/kg body weight) was injected into the abomasal lumen during surgery. Jugular venous blood samples for determination of serum d-xylose concentration were periodically obtained. Time to maximal serum D-xylose concentration (T(max-model)) was pharmacokinetically determined. RESULTS: Abomasal emptying rate was significantly (P<0.05) faster in group E (T(max-model)=182+/-69 min; mean+/-SD) than in group C cows (T(max-model)=237+/ 64 min). Abomasal emptying rate was similar in group D (T(max-model)=196+/-47 min) and group C. Both treatments improved postoperative milk yield within 1 day after surgery. CONCLUSION: Preoperative injection of erythromycin (10 mg/kg IM) is an effective method for ameliorating postoperative abomasal hypomotility in cows with AV. CLINICAL RELEVANCE: Parenteral erythromycin can be recommended for preoperative treatment of cows with AV. PMID- 19134104 TI - Assessment of humeral length in dogs after repair of Salter-Harris type IV fracture of the lateral part of the humeral condyle. AB - OBJECTIVE: To evaluate the effect of fracture and subsequent repair on future bone growth of the humerus after Salter-Harris type IV fracture of the lateral part of the humeral condyle (LPHC). STUDY DESIGN: Prospective study. ANIMALS: Dogs (n=11). METHODS: Dogs that had LPHC fracture and an open distal humeral physis repaired (1992-2006) were re-examined and radiographed at >or=12 months of age and humeral length was measured. RESULT: Measurements from 11 dogs showed a significant (P=.02) increase in length of the humeral diaphysis of the affected leg compared with that of the intact limb (median, 1.2%; range, 1.3-3.4%). Condylar deformity secondary to growth disturbance was not observed. CONCLUSION: Shortening or growth deformity was not observed after fracture and repair even if a transcondylar screw was placed through the distal humeral growth plate. A mild overgrowth of the humeral diaphysis was observed, although likely considered clinically unimportant. CLINICAL RELEVANCE: Fracture of the LPHC and subsequent repair in dogs >3 months of age do not impair growth of the humeral diaphysis. A transcondylar humeral screw placed through the humeral physis will not result in shortening of the humeral diaphysis. Implant removal to allow for further growth is therefore not indicated. PMID- 19134105 TI - Clinical use of recombinant human bone morphogenic protein-2 in a whooping crane (Grus americana). AB - OBJECTIVE: To report use of recombinant human bone morphogenetic protein-2 (rhBMP 2) as adjunctive therapy for treatment of a comminuted, open, proximal humeral fracture in an avian species. STUDY DESIGN: Clinical report. ANIMALS: A 3.5-month old male whooping crane (Grus americana). METHODS: An open, severely comminuted humeral facture was stabilized with an intramuscular (IM) pin/type IA external skeletal fixator with tie-in configuration. rhBMP-2 was applied in a calcium phosphate matrix (CPM) paste directly to the fracture site as a bone graft substitute. Radiographic evidence of bone healing was monitored for 14 weeks. RESULTS: Substantial bony callus was evident at 4 weeks and at 8 weeks there was bridging callus with obvious bony remodeling. The fixation was destabilized at 9 weeks by IM pin removal, bone healing progressed and the fixator was removed at 11 weeks. By 14 weeks both cortices had been re-established with continued callus remodeling evident. CONCLUSION: rhBMP-2, applied in a CPM paste, was used as a bone graft substitute in the treatment of a comminuted, open humeral fracture in a whooping crane. CLINICAL RELEVANCE: Use of rhBMP-2/CPM should be considered in treatment of avian fractures. PMID- 19134106 TI - Analysis of sodium carboxymethylcellulose administration and related factors associated with postoperative colic and survival in horses with small intestinal disease. AB - OBJECTIVE: To analyze the effect of the intraoperative use of sodium carboxymethylcellulose (CBMC) and related perioperative factors on postoperative colic and survival in horses that had abdominal surgery for colic. STUDY DESIGN: Retrospective study. ANIMALS: Horses (n=203) that had surgery for small intestinal disease; 33 horses had intraoperative administration of CBMC. METHODS: Information was obtained from medical records for 170 horses that had surgery for colic before use of CBMC and 33 horses that had intraoperative CBMC. Kaplan-Meier survival curves were used to estimate median survival time and a Cox proportional hazards model was used to estimate the hazard ratio for the effect of CBMC and other perioperative variables on survival. RESULTS: Seventy-five percent of horses administered CBMC survived to 180 days, whereas 75% of untreated horses survived 8 days (median survival time=18 days). Horses not administered CBMC were twice as likely to die compared with horses administered CBMC. Horses that had postoperative ileus (POI) were 1.4 times more likely to die than horses without ileus. Similarly, horses with signs of colic after surgery were 1.3 times more likely to die than horses without postoperative signs of colic. CONCLUSIONS: CBMC administration is seemingly protective against death and prolongs survival when used intraoperatively in horses with small intestine disease, particularly horses with postoperative colic or POI. Both POI and colic increased risk of death after surgery. CLINICAL RELEVANCE: Intraoperative administration of CBMC in horses that have surgery for small intestinal disease may improve survival, possibly by reducing early adhesion formation. PMID- 19134107 TI - Evaluation of continuous infusion of lidocaine on gastrointestinal tract function in normal horses. AB - OBJECTIVE: To determine the effect of continuous infusion of lidocaine on fecal transit time in normal horses. STUDY DESIGN: Experimental randomized cross-over study. ANIMALS: Healthy horses (n=6). METHODS: Barium-filled microspheres were administered to horses by nasogastric intubation and feces were collected every 2 hours for 4 days. A bolus of 2% lidocaine (1.3 mg/kg) was administered randomly, followed by a continuous infusion of lidocaine (0.05 mg/kg/min) for 3 days or an equivalent volume of saline. The washout period was 10 days. Variables assessed included defecation frequency, weight of feces produced, intestinal transit time (number of microspheres observed on radiographs), fecal moisture content, borborygmus score, heart and respiratory rate, and signs of lidocaine toxicity (e.g., ataxia, CNS depression). RESULTS: During the first 24 hours of lidocaine administration, mean (+/-SD) fecal output (10.8+/-6.9 kg) was decreased compared with controls (15+/-4.9 kg). Mean (+/-SEM) time for passing 50% of the barium filled microspheres was shorter in controls (42+/-1.13 hours) compared with the lidocaine group (50+/-1.32 hours). CONCLUSIONS: Continuous infusion of lidocaine increases the transit time of feces in normal horses. CLINICAL RELEVANCE: Clinicians need to be aware of the effects of using a continuous infusion of lidocaine on the transit time of feces in normal horses, with a potential for exacerbating those effects when combined with drugs that decrease motility and in horses with medical colic (e.g., impaction) or where a diagnosis has not been made. PMID- 19134108 TI - Mandibular osteodistraction for correction of deep bite class II malocclusion in a horse. AB - OBJECTIVE: To describe a technique for, and outcome after, mandibular osteodistraction in the horse. STUDY DESIGN: Clinical report. ANIMALS: Warmblood horse. METHODS: A half ring external fixator was applied on both sides of an osteotomy site performed on the mandible of a colt. A bite plate was placed on the upper incisors creating occlusion between lower and upper jaw. After a 5-day latency period, distraction was applied (1 mm/day) until the overjet was judged normal. RESULTS: Mandibular elongation and correction of brachygnathia was obtained without major complications. Six months after the procedure the overjet reduction was considered stable. CONCLUSIONS: Mandibular osteodistraction can be considered for treatment of severe brachygnathia in yearlings. CLINICAL RELEVANCE: Distraction osteogenesis has the advantage of progressive elongation of the mandible, allowing concurrent bone remodeling and soft tissue adaptation. Severe mandibular incisor malocclusion in horses outside the maximal growth phase can be corrected using this technique. PMID- 19134109 TI - Biomechanical comparison of six suture configurations using a large diameter polyester prosthesis in the muscular process of the equine arytenoid cartilage. AB - OBJECTIVE: To compare biomechanical properties of 6 suture configurations using a large diameter polyester prosthesis in the muscular process (MP) of the arytenoid cartilage and to determine failure mode. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Cadaveric equine larynges (n=121). METHODS: Suture configurations (4 single, 2 double) were inserted, and then constructs were tested in a single cycle to failure at a 100 mm/min distraction. Load deformation curves were generated to assess the biomechanical properties of each construct. A 1-way ANOVA was used to compare the mean differences in construct failure force, cricoarytenoid joint (CAJ) disarticulation force, and energy stored at failure. A 2-sample t-test was used to compare single versus double suture patterns and a Fisher's exact test was used to compare failure mode. RESULTS: Both construct and CAJ failure force were significantly greater (P<.05) for double suture patterns compared with single suture patterns; however, there were no significant differences in energy stored at construct failure. Failure at the MP accounted for >or=50% of construct failures for 3 of the single suture patterns and 1 of the double suture patterns tested. The remaining 2 patterns had an increased frequency of clamp failures as well as failure of the cricoid cartilage. CONCLUSIONS: Sutures that sufficiently engage the spine of the MP alone or in conjunction with a second suture were found to be biomechanically superior. CLINICAL RELEVANCE: Engaging the spine of the MP appears to result in the most biomechanically sound laryngeal construct. PMID- 19134110 TI - In vitro model for testing novel implants for equine laryngoplasty. AB - OBJECTIVE: To develop an in vitro laryngeal model to mimic airflow and pressures experienced by horses at maximal exercise with which to test laryngoplasty techniques. STUDY DESIGN: Randomized complete block. SAMPLE POPULATION: Cadaveric equine larynges (n=10). METHODS: Equine larynges were collected at necropsy and a bilateral prosthetic laryngoplasty suture was placed with #5 Fiberwire suture to achieve bilateral maximal arytenoid abduction. Each larynx was positioned in a flow chamber and subjected to static flow and dynamic flow cycling at 2 Hz. Tracheal pressure and flow, and pressure within the flow chamber were recorded at a sampling frequency of 500 Hz. Data obtained were compared with the published physiologic values for horses exercising at maximal exercise. RESULTS: Under static flow conditions, the testing system produced inspiratory tracheal pressures (mean+/-SEM) of -33.0+/-0.98 mm Hg at a flow of 54.48+/-1.8 L/s. Pressure in the flow chamber was -8.1+/-2.2 mm Hg producing a translaryngeal impedance of 0.56+/-0.15 mm Hg/L/s. Under dynamic conditions, cycling flow and pressure were reproduced at a frequency of 2 Hz, the peak inspiratory (mean+/ SEM) pharyngeal and tracheal pressures across all larynges were -8.85+/-2.5 and 35.54+/-1.6 mm Hg, respectively. Peak inspiratory flow was 51.65+/-2.3 L/s and impedance was 0.57+/-0.06 mm Hg/L/s. CONCLUSIONS: The model produced inspiratory pressures similar to those in horses at maximal exercise when airflows experienced at exercise were used. CLINICAL RELEVANCE: This model will allow testing of multiple novel techniques and may facilitate development of improved techniques for prosthetic laryngoplasty. PMID- 19134111 TI - Epidural idiopathic sterile pyogranulomatous inflammation causing spinal cord compressive injury in five Miniature Dachshunds. AB - OBJECTIVE: To characterize the clinical signs, diagnostic and surgical findings, and outcome of dogs with idiopathic sterile pyogranulomatous inflammation (ISP) of epidural fat causing spinal cord compression. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=5). METHODS: Dogs with epidural ISP (2002-2006) were identified retrospectively. Inclusion criteria were neurologic examination, myelography, and definitive diagnosis of ISP confirmed by surgery and histopathologic examination of epidural spinal cord compressive tissue. RESULTS: The most common clinical sign was paraparesis/paraplegia. No abnormalities were detected by laboratory testing or survey spine radiographs. On myelography, extradural spinal cord compressions were focal (dogs 1, 3, and 5) or multifocal (dogs 2 and 4). Surgical decompression of the spinal cord was completed by hemilaminectomy. Epidural fat collected surgically had pyogranulomatous inflammation of unknown cause and was histologically similar to subcutaneous ISP. All dogs had good long-term neurologic outcome (10-45 months follow-up). Some dogs had episodes of ISP at other sites before or after surgical treatment of epidural ISP, suggesting there may be a systemic form of ISP. CONCLUSION: Epidural ISP may cause a spinal cord compressive lesion in Miniature Dachshunds, which can be treated by surgical decompression of the spinal cord with or without administration of adjunctive steroids. CLINICAL RELEVANCE: Epidural ISP should be considered as a possible cause of thoracolumbar myelopathy for Miniature Dachshunds. PMID- 19134112 TI - Novel components of the Apicomplexan moving junction reveal conserved and coccidia-restricted elements. AB - Apicomplexan parasites generally invade their host cells by anchoring the parasite to the host membrane through a structure called the moving junction (MJ). This MJ is also believed to sieve host proteins from the nascent parasitophorous vacuole membrane, which likely protects the pathogen from lysosomal destruction. Previously identified constituents of the Toxoplasma MJ have orthologues in Plasmodium, indicating a conserved structure throughout the Apicomplexa. We report here two novel MJ proteins, RON5 and RON8. While RON5 is conserved in Plasmodium, RON8 appears restricted to the coccidia. RON8, which is likely essential, co-immunoprecipitates RON5 and known MJ proteins from extracellular parasites, indicating that a preformed complex exists within the parasites. Upon secretion, we show that RON8 within the MJ localizes to the cytoplasmic face of the host plasma membrane. To examine interactions between RON8 and the host cell, we expressed RON8 in mammalian cells and show that it targets to its site of action at the periphery in a manner dependent on the C terminal portion of the protein. The discovery of RON5 and RON8 provides new insight into conserved and unique elements of the MJ, furthering our understanding of how the MJ contributes to the intricate mechanism of Apicomplexan invasion. PMID- 19134114 TI - Mutualism versus pathogenesis: the give-and-take in plant-bacteria interactions. AB - Pathogenic bacteria and mutualistic rhizobia are able to invade and establish chronic infections within their host plants. The success of these plant-bacteria interactions requires evasion of the plant innate immunity by either avoiding recognition or by suppressing host defences. The primary plant innate immunity is triggered upon recognition of common microbe-associated molecular patterns. Different studies reveal striking similarities between the molecular bases underlying the perception of rhizobial nodulation factors and microbe-associated molecular patterns from plant pathogens. However, in contrast to general elicitors, nodulation factors can control plant defences when recognized by their cognate legumes. Nevertheless, in response to rhizobial infection, legumes show transient or local defence-like responses suggesting that Rhizobium is perceived as an intruder although the plant immunity is controlled. Whether these responses are involved in limiting the number of infections or whether they are required for the progression of the interaction is not yet clear. Further similarities in both plant-pathogen and Rhizobium-legume associations are factors such as surface polysaccharides, quorum sensing signals and secreted proteins, which play important roles in modulating plant defence responses and determining the outcome of the interactions. PMID- 19134113 TI - The ex vivo response of human intestinal mucosa to enteropathogenic Escherichia coli infection. AB - In vitro organ culture (IVOC) represents a gold standard model to study enteropathogenic E. coli (EPEC) infection of human intestinal mucosa. However, the optimal examination of the bacterial-host cell interaction requires a directional epithelial exposure, without serosal or cut surface stimulation. A polarized IVOC system (pIVOC) was developed in order to overcome such limitations: apical EPEC infection produced negligible bacterial leakage via biopsy edges, resulted in enhanced colonization compared with standard IVOC, and showed evidence of bacterial detachment, as in natural rabbit EPEC infections. Examination of mucosal innate immune responses in pIVOC showed both interleukin (IL)-8 mRNA and protein levels were significantly increased after apical EPEC infection. Increased IL-8 levels mainly depended on flagellin expression as fliC negative EPEC did not elicit a significant IL-8 response despite increased mucosal colonization compared with wild-type EPEC. In addition, apical application of purified flagella significantly increased IL-8 protein levels over non-infected controls. Immunofluorescence staining of EPEC-infected small intestinal biopsies revealed apical and basolateral distribution of Toll-like receptor (TLR) 5 on epithelium, suggesting that EPEC can trigger mucosal IL-8 responses by apical flagellin/TLR5 interaction ex vivo and does not require access to the basolateral membrane as postulated in cell culture models. PMID- 19134115 TI - Mathematical and computational approaches can complement experimental studies of host-pathogen interactions. AB - In addition to traditional and novel experimental approaches to study host pathogen interactions, mathematical and computer modelling have recently been applied to address open questions in this area. These modelling tools not only offer an additional avenue for exploring disease dynamics at multiple biological scales, but also complement and extend knowledge gained via experimental tools. In this review, we outline four examples where modelling has complemented current experimental techniques in a way that can or has already pushed our knowledge of host-pathogen dynamics forward. Two of the modelling approaches presented go hand in hand with articles in this issue exploring fluorescence resonance energy transfer and two-photon intravital microscopy. Two others explore virtual or 'in silico' deletion and depletion as well as a new method to understand and guide studies in genetic epidemiology. In each of these examples, the complementary nature of modelling and experiment is discussed. We further note that multi-scale modelling may allow us to integrate information across length (molecular, cellular, tissue, organism, population) and time (e.g. seconds to lifetimes). In sum, when combined, these compatible approaches offer new opportunities for understanding host-pathogen interactions. PMID- 19134116 TI - Candida albicans actively modulates intracellular membrane trafficking in mouse macrophage phagosomes. AB - The intracellular trafficking/survival strategies of the opportunistic human pathogen Candida albicans are poorly understood. Here we investigated the infection of RAW264.7 macrophages with a virulent wild-type (WT) filamentous C. albicans strain and a hyphal signalling-defective mutant (efg1Delta/cph1Delta). A comparative analysis of the acquisition by phagosomes of actin, and of early/late endocytic organelles markers of the different fungal strains was performed and related to Candida's survival inside macrophages. Our results show that both fungal strains have evolved a similar mechanism to subvert the 'lysosomal' system, as seen by the inhibition of the phagosome fusion with compartments enriched in the lysobisphosphatidic acid and the vATPase, and thereby the acquisition of a low pH from the outset of infection. Besides, the virulent WT strain displayed additional specific survival strategies to prevent its targeting to compartmentsdisplaying late endosomal/lysosomal features, such as induction of active recycling out of phagosomes of the lysosomal membrane protein LAMP-1, the lysosomal protease cathepsin D and preinternalized colloidal gold. Finally, both virulent and efg1Delta/cph1Delta mutant fungal strains actively suppressed the production of macrophage nitric oxide (NO), although their cell wall extracts were potent inducers of NO. PMID- 19134117 TI - Human TLR3 recognizes dengue virus and modulates viral replication in vitro. AB - The elicitation of large amount inflammatory cytokine in serum has been developed as the cause of the plasma leakage in dengue fever (DF)/dengue haemorrhagic fever (DHF) infection. Virus recognition in innate immunity is the key. The Toll-like receptors (TLRs) play an important role in pathogen recognition towards cytokine induction among several viruses; however, the role of TLRs on innate immune recognition against DENV remains unclear. This study aims at the interaction between dengue virus (DENV) and human TLRs at the incipient stage of infection in vitro. Our experiment reveals that stably expression of TLR3, 7, 8 on HEK293 enables IL-8 secretion after DENV recognition. By the model of human monocytic cells U937, we demonstrated the trigger of IL-8 after viral recognition of human monocytic cell is primary through TLR3 following endosomal acidification. Silencing of TLR3 in U937 cells significantly blocks the DENV-induced IL-8 production. Besides, the interaction is further corroborated by colocalization of TLR3 and DENV RNA upon DENV internalization. Furthermore, in this study we found the expression of TLR3 can mediate strong IFN-alpha/beta release and inhibit DENV viral replication significantly, thus limit the cytopathic effect. PMID- 19134118 TI - Innate immune responses to Mycobacterium ulcerans via toll-like receptors and dectin-1 in human keratinocytes. AB - Mycobacterium ulcerans (MU), an environmental pathogen, causes Buruli ulcer, a severe skin disease. We hypothesized that epidermal keratinocytes might not be a simple barrier, but play a role during MU infection through pattern-recognition receptors expressed in keratinocytes. We found that keratinocyte Toll-like receptors (TLRs) 2 and 4 and Dectin-1 actively participate in the innate immune response to MU, which includes the internalization of bacteria, the production of reactive oxygen species (ROS), and the expression of chemokines and LL-37. Human keratinocytes constitutively expressed TLRs 2 and 4 and induced Dectin-1 in response to MU. Exposing keratinocytes to MU resulted in rapid ROS production, which in turn contributed to the mRNA and protein expression of LL-37. In addition, TLR2, Dectin-1 and, to an extent, TLR4 are essential for the MU mediated expression of CXCL8, CCL2 and LL-37 in keratinocytes. Furthermore, confocal analysis showed that the Dectin-1 is necessary for keratinocytes to internalize bacilli. Importantly, blockade of ROS and LL-37 significantly increased the intracellular MU growth in keratinocytes, suggesting an important role of these mediators for cutaneous innate immune responses. Our results demonstrate that TLR2, TLR4 and Dectin-1 actively sense, internalize and respond in an innate way to MU in human epidermal keratinocytes. PMID- 19134119 TI - Identification and characterization of Ibe, a novel type III effector protein of A/E pathogens targeting human IQGAP1. AB - Enteropathogenic Escherichia coli (EPEC), atypical enteropathogenic Escherichia coli (ATEC) and enterohemorrhagic Escherichia coli (EHEC) belong to the family of attaching and effacing (A/E) pathogens. Pathogenicity is mediated by subversion of host cell functions involving type III secretion system (TTSS)-dependent effector proteins. In this study, we have identified and characterized a novel TTSS-dependent effector protein encoded at the 5'-end of the locus of enterocyte effacement (LEE) pathogenicity island (PAI) of ATEC strain 3431-4/86 (O8:H(-)). Using affinity purification we identified IQGAP1, a scaffolding protein involved in the regulation of the actin cytoskeleton, as a putative host cell target. Accordingly, we termed the novel effector protein 'Ibe' for IQGAP1-binding effector. The interaction of Ibe and IQGAP1 was confirmed by co immunoprecipitation from ATEC-infected cells and immunofluorescence analysis, which revealed colocalization of Ibe and IQGAP1 in ATEC-induced pedestals and actin-rich membrane ruffles. This suggests that the putative effector function of Ibe is mediated via IQGAP1. The Ibe-independent recruitment of IQGAP1 to ATEC induced pedestals implies a general role for IQGAP1 in the subversion of host cell functions during infection. Homologues of the novel effector Ibe are widely distributed among EPEC, ATEC and EHEC strains but are not necessarily genetically linked to the LEE as they have occasionally also been found to be encoded within lambdoid prophages. PMID- 19134120 TI - Rickettsial outer-membrane protein B (rOmpB) mediates bacterial invasion through Ku70 in an actin, c-Cbl, clathrin and caveolin 2-dependent manner. AB - Rickettsia conorii, an obligate intracellular tick-borne pathogen and the causative agent of Mediterranean spotted fever, binds to and invades non phagocytic mammalian cells. Previous work identified Ku70 as a mammalian receptor involved in the invasion process and identified the rickettsial autotransporter protein, rOmpB, as a ligand; however, little is known about the role of Ku70 rOmpB interactions in the bacterial invasion process. Using an Escherichia coli heterologous expression system, we show here that rOmpB mediates attachment to mammalian cells and entry in a Ku70-dependent process. A purified recombinant peptide corresponding to the rOmpB passenger domain interacts with Ku70 and serves as a competitive inhibitor of adherence. We observe that rOmpB-mediated infection culminates in actin recruitment at the bacterial foci, and that this entry process relies in part on actin polymerization likely imparted through protein tyrosine kinase and phosphoinositide 3-kinase-dependent activities and microtubule stability. Small-interfering RNA studies targeting components of the endocytic pathway reveal that entry by rOmpB is dependent on c-Cbl, clathrin and caveolin-2. Together, these results illustrate that rOmpB is sufficient to mediate Ku70-dependent invasion of mammalian cells and that clathrin- and caveolin-dependent endocytic events likely contribute to the internalization process. PMID- 19134121 TI - Uropathogenic Escherichia coli AL511 requires flagellum to enter renal collecting duct cells. AB - Escherichia coli is the leading cause of urinary tract infections, but the mechanisms governing renal colonization by this bacterium remain poorly understood. We investigated the ability of 13 E. coli strains isolated from the urine of patients with pyelonephritis and cystitis and normal stools to invade collecting duct cells, which constitute the first epithelium encountered by bacteria ascending from the bladder. The AL511 clinical isolate adhered to mouse collecting duct mpkCCD(cl4) cells, used as a model of renal cell invasion, and was able to enter and persist within these cells. Previous studies have shown that bacterial flagella play an important role in host urinary tract colonization, but the role of flagella in the interaction of E. coli with renal epithelial cells remains unclear. An analysis of the ability of E. coli AL511 mutants to invade renal cells showed that flagellin played a key role in bacterial entry. Both flagellum filament assembly and the motor proteins MotA and MotB appeared to be required for E. coli AL511 uptake into collecting duct cells. These findings indicate that pyelonephritis-associated E. coli strains may invade renal collecting duct cells and that flagellin may act as an invasin in this process. PMID- 19134123 TI - Formulation effects of topical emulsions on transdermal and dermal delivery. AB - It has been recognized that the vehicle in which a permeant is applied to the skin has a distinctive effect on the dermal and transdermal delivery of active ingredients. The cutaneous and percutaneous absorptions can be enhanced, e.g. by an increase in thermodynamic activity, supersaturation and penetration modifiers. Furthermore, dermal and transdermal delivery can be influenced by the interactions that may occur between the vehicle and the skin on the one hand, and interactions between the active ingredient and the skin on the other hand. Emulsions are widely used as cosmetic and pharmaceutical formulations because of their excellent solubilizing capacities for lipophilic and hydrophilic active ingredients and application acceptability. This review focuses, in particular, on the effect of emulsions on the dermal and transdermal delivery of active ingredients. It is shown that the type of emulsion (w/o vs. o/w emulsion), the droplet size, the emollient, the emulsifier as well as the surfactant organization (micelles, lyotropic liquid crystals) in the emulsion may affect the cutaneous and percutaneous absorption. Examples substantiate the fact that emulsion constituents such as emollients and emulsifiers should be selected carefully for optimal efficiency of the formulation. Moreover, to understand the influence of emulsion on dermal and transdermal delivery, the physicochemical properties of the formulation after application are considered. PMID- 19134122 TI - Live cell fluorescence microscopy to study microbial pathogenesis. AB - Advances in microscopy and fluorescent probes provide new insight into the nanometer-scale biochemistry governing the interactions between eukaryotic cells and pathogens. When combined with mathematical modelling, these new technologies hold the promise of qualitative, quantitative and predictive descriptions of these pathways. Using the light microscope to study the spatial and temporal relationships between pathogens, host cells and their respective biochemical machinery requires an appreciation for how fluorescent probes and imaging devices function. This review summarizes how live cell fluorescence microscopy with common instruments can provide quantitative insight into the cellular and molecular functions of hosts and pathogens. PMID- 19134124 TI - Human synthetic sebum formulation and stability under conditions of use and storage. AB - The human skin surface and hair are generally coated with a thin film of liquid phase sebaceous lipids. This surface lipid film contributes to the cosmetic properties of the skin. Synthetic sebum has been used for studies on properties of skin and hair. However, there has been no standardized formulation of synthetic sebum and many of the synthetic sebum formulations that have been used do not closely resemble actual sebum. In this study, a formulation for a standardized and inexpensive synthetic sebum is proposed, and the chemical stability of this lipid mixture is demonstrated under conditions of use and storage. The proposed synthetic sebum consists of 17% fatty acid, 44.7% triglyceride, 25% wax monoester (jojoba oil) and 12.4% squalene. This lipid mixture takes up approximately 6% of its weight in water when equilibrated in an atmosphere saturated with water vapour. It is stable on exposure to the atmosphere at 32 degrees C for at least 48 h, and it is also stable on storage at 4 or -20 degrees C, either dry or in chloroform : methanol solution for at least 6 months. This synthetic sebum could be useful in studies on cosmetic properties of the skin surface or hair, on penetration of chemicals into the skin or in development of standardized tests of laundry detergent performance. PMID- 19134125 TI - Development of a minimally invasive epidermal abrasion device for clinical skin sampling and its applications in molecular biology. AB - A new abrasion tool (US patent US7087063 B2) has been developed for collecting skin epidermal samples. This device includes a central shaft that holds the probe in a split chuck. Of the variety of probe designs tested, the laser-cut hollow tube (HT) probe abraded the basal layer of the epidermis most consistently, resulting in representative epidermal skin samples. Compared with traditional clinical methods, the abrasion method allows for high-throughput epidermal skin collection with minimal invasiveness to the volunteer subjects. A large number of abrasion samples have been collected in various clinical studies with no adverse effects observed. Epidermal abrasion, when used appropriately and with the optimized probes, can yield high quality tissue samples that are representative of the epidermis. A sufficient quantity of RNA and protein can be obtained for many subsequent molecular and biochemical applications. Because of its minimal invasiveness and high-throughput nature, the abrasion method can be a valuable tool used to investigate the efficacy of topical applications of skin care products. PMID- 19134126 TI - Comparison of clinical efficacies of sodium ascorbyl phosphate, retinol and their combination in acne treatment. AB - Acne vulgaris impairs the appearance of an individual and causes psychological irritation. Inflammatory acne lesion is caused by multifactor incorporates in each step of acne pathogenesis. In an attempt to archive inflammatory lesion treatment with the promise of prevention of acne vulgaris, randomized and double blind studies on the comparison of the efficacies of topical formulations containing 5% sodium ascorbyl phosphate (SAP) and 0.2% retinol, separately as well as in combination application, were conducted. The resulting data showed that SAP reduced the inflammatory lesion by 20.14% and 48.82% within 4 and 8 weeks respectively. Application of the formulation containing retinol slightly improved the treatment efficacy as the lesion reduced by 21.79% and 49.50% after 4 and 8 weeks respectively. The combination treatment significantly reduced the inflammatory lesion by 29.28% after 4 weeks and 63.10% after 8 weeks of application. The most effective treatment was by using the combination of 5% SAP and 0.2% retinol, which incorporated the synergistic effects on lipid peroxidation and sebaceous gland function in addition to the enhancement of SAP permeability by the desquamation of stratum corneum influenced by retinol, keratin plug removal and anti-inflammatory effect of retinol. This study promises for the development of cosmetic products to overcome aesthetic and psychological problems caused by acne vulgaris. PMID- 19134127 TI - Manipulation of body odour alters men's self-confidence and judgements of their visual attractiveness by women. AB - Human body odour is important in modulating self-perception and interactions between individuals. Artificial fragrances have been used for thousands of years to manipulate personal odour, but the nature and extent of influences on person perception are relatively unexplored. Here we test the effects of a double-blind manipulation of personal odour on self-confidence and behaviour. We gave to male participants either an aerosol spray containing a formulation of fragrance and antimicrobial agents or an otherwise identical spray that lacked these active ingredients. Over several days, we found effects between treatment groups on psychometric self-confidence and self-perceived attractiveness. Furthermore, although there was no difference between groups in mean attractiveness ratings of men's photographs by a female panel, the same women judged men using the active spray as more attractive in video-clips, suggesting a behavioural difference between the groups. Attractiveness of an individual male's non-verbal behaviour, independent of structural facial features, was predicted by the men's self reported proclivity towards the provided deodorant. Our results demonstrate the pervasive influence of personal odour on self-perception, and how this can extend to impressions on others even when these impressions are formed in the absence of odour cues. PMID- 19134129 TI - Spectral uniformity: a new index of broad spectrum (UVA) protection. AB - Despite the wide acceptance of in vitro assay for the determination of the spectral absorbance of a sunscreen product, we find that different authorities are recommending different summary metrics of this spectral profile to express the UVA or broad spectrum, protection offered by topical sunscreens. Regrettably, the situation that now prevails is not ideal and the option for rationalizing the situation in the interests of consumer-focussed international harmonization is either the adoption of one of the existing metrics or universal acceptance of a new metric for expressing broad spectrum (UVA) protection. There may be greater harmony in adopting the second proposal and so I introduce here a new metric, which I term the spectral uniformity index, for consideration by the international sunscreen community. It is shown that this new index is aligned to the fundamental requirement of a topical sunscreen to provide a flat spectral profile, is simple to calculate and conceptually easy to understand, and correlates strongly with existing indices. PMID- 19134128 TI - Salvia somalensis essential oil as a potential cosmetic ingredient: solvent-free microwave extraction, hydrodistillation, GC-MS analysis, odour evaluation and in vitro cytotoxicity assays. AB - Salvia somalensis Vatke, a wild sage native of Somalia, has been studied with the aim of assessing the potential cosmetic application of its essential oil, recovered from fresh aerial parts by solvent-free microwave extraction - SFME. To evaluate the efficiency and reliability of this eco-friendly procedure, the recovery of the essential oil was also processed by conventional hydrodistillation (HD) and the results compared. The essential oils obtained by both SFME and HD were analysed by gas chromatography-mass spectrometry using apolar and polar capillary columns. The essential oil recovered by SFME was submitted to an odour evaluation that revealed peculiar olfactive characteristics interesting in alcoholic male perfumery and body detergents.In vitro cytotoxicity assays were carried out using NCTC 2544 human keratinocytes as target cells. The oil displayed slight cytotoxic effects, which were three orders of magnitude lower than those found for sodium dodecyl sulphate positive control. The promising results in terms of chemical composition, scent and safety seem to indicate this essential oil as an interesting potential functional ingredient useful in a cosmetic context. PMID- 19134145 TI - Honorary Membership of International Xenotransplantation Association conferred upon Professor Ian F C McKenzie (September 18, 2007). PMID- 19134160 TI - Selective rejection of porcine islet xenografts by macrophages. AB - BACKGROUND: Our previous study has shown that porcine antigen-primed and CD4+ T cell-activated macrophages are capable of recognition and rejection of porcine xenografts after adoptive transfer. However, whether this is an absolute xenograft specific rejection remains to be confirmed. METHODS: Mouse islet allografts and neonatal porcine islet cell cluster (NICC) xenografts were admixed and transplanted under the left kidney capsule, and NICC xenografts alone were transplanted under the right kidney capsule of strepotozotocin-induced diabetic NOD-SCID mice. After achievement of normoglycemia, the NOD-SCID recipients were transferred with macrophages purified from NICC transplant NOD-SCID mice reconstituted with CD4+ T cells. Five weeks after macrophage transfer the left kidney with the admixed grafts were removed. Graft survival and function following macrophage transfer was assessed by blood glucose measurement and immunohistochemistry. RESULTS AND CONCLUSIONS: Adoptive transfer with activated macrophages did not affect the normalized blood glucose levels in NOD-SCID recipients of admixed grafts until left nephrectomy 5 weeks post-macrophage transfer. Insulin-positive and porcine C-peptide-negative mouse islets were detected in the admixed grafts. The surviving mouse islets in the admixed grafts were surrounded but not infiltrated by macrophages. The nephrectomized recipients demonstrated sustained hyperglycemia and completely destroyed NICC xenografts in their remaining right kidneys 8 weeks after macrophage transfer. Taken together, these data provide direct evidence of porcine islet xenograft specific rejection by activated macrophages. PMID- 19134161 TI - High antigen levels do not preclude B-cell tolerance induction to alpha1,3-Gal via mixed chimerism. AB - BACKGROUND: Studies of bone marrow transplantation (BMT) from wild-type mice or rats to alpha1,3-galactosyltransferase (GalT) knockout mice have demonstrated that induction of mixed chimerism tolerizes not only T cells, but also natural antibody-producing B cells, even across xenogeneic barriers. Given that rodent cells express lower levels of the alphaGal epitope than the more clinically relevant porcine species, the consequences of exposure to cells expressing high levels of alphaGal on the ability to induce B-cell tolerance are unknown. METHODS: The effects on chimerism and anti-alphaGal B-cell tolerance of an i.p. injection of 10(9) porcine RBC were evaluated in GalT knockout mice receiving wild-type allogeneic BMT after non-myeloablative conditioning with T-cell depleting monoclonal antibodies, thymic irradiation, and low-dose total body irradiation. RESULTS: Achievement of mixed chimerism and tolerance of anti alphaGal-producing B cells was not affected by exposure to high-density alphaGal at the time of BMT. The absence of induced anti-alphaGal or anti-pig antibody responses in conditioned control mice suggested that the B-cell xeno-response to pig is T-cell-dependent. CONCLUSION: High alphaGal density on pig cells might not preclude the ability to achieve tolerance of pre-existing alphaGal-reactive human B cells via induction of mixed chimerism. This strategy has the potential to induce B-cell tolerance to non-alphaGal epitopes, against which natural antibodies have been found in the sera of healthy humans. PMID- 19134162 TI - Porcine cells express more than one functional ligand for the human lymphocyte activating receptor NKG2D. AB - BACKGROUND: Xenotransplantation could ameliorate the severe shortage of donor organs. The initial results of transplantation from genetically-modified pig donors to primate recipients suggest that hyperacute rejection can be overcome, but thrombotic microangiopathy and the human anti-pig cellular immune response remain as significant impediments to successful clinical xenotransplantation. NKG2D is an activating immunoreceptor found on human natural killer (HuNK) cells, CD8(+) and gammadelta T cells. Signaling through NKG2D mediates cytotoxicity and cytokine secretion by NK cells and co-stimulation of T cells. METHODS: Chinese hamster ovary P (CHOP) cells were transfected with human NKG2D and used in cell cell binding studies with porcine epithelial, and endothelial cell lines. Soluble recombinant NKG2D-Fc was used to stain various porcine cells and tissues to indicate ligand expression. Porcine cells were used as targets in cytotoxicity assays with the HuNK cell lines NKL and YT, with and without enzymatic removal of pULBP1 and antibody blockade of NKG2D signaling. RESULTS AND CONCLUSIONS: In this study, we demonstrate the expression of ligands for human NKG2D on porcine cell lines of endothelial and epithelial origin, islet cell clusters and rejecting kidney. HuNK cells were activated to kill pig cells expressing NKG2D ligands, and cytotoxicity was inhibited by antibody blockade of NKG2D. A previous study identified pULBP1 as the principal ligand for human NKG2D on pig aortic endothelial cells. In the current study, renal epithelial and intestinal endothelial cells each expressed high surface levels of pULBP1, but binding of soluble recombinant NKG2D and NKG2D-dependent cytotoxicity against these cells persisted after the enzymatic removal of pULBP1, strongly suggesting the presence of at least one additional functional ligand for human NKG2D in these cell types. PMID- 19134163 TI - Prolonged survival of pig islets xenograft by adenovirus-mediated expression of either the membrane-bound human FasL or the human decoy Fas antigen gene. AB - BACKGROUND: Pig islets are considered an attractive alternative treatment for patients with Type 1 diabetes. However, pig islet xenografts, transplanted into non-human primates, are directly rejected by cell-mediated processes. We have previously reported that cell-mediated xenograft-rejections, and especially human CD8(+) cytotoxic T lymphocytes (CTL)-mediated cytotoxicity, are highly detrimental to pig xenograft cells. Moreover, we have explored novel strategies for the prevention of CTL killing by overexpression of either human decoy Fas antigen or membrane-bound human FasL in pig endothelial cells. In this study, we assessed the cytoprotective effects of these molecules for pig islets both in vitro and in vivo. MATERIALS AND METHODS: Pig islets were freshly isolated by modified Ricordi's methods. Subsequently, these islets were transfected with an adenoviral expression vector containing the DNA fragments of either membrane bound human FasL or human decoy Fas. Transfected islets were transplanted into preimmunized diabetic rats under the kidney capsule. Control pig islets (i.e., MOCK), which were transfected with an adenoviral expression vector containing only the enhanced green fluorescent protein gene, were also transplanted. RESULTS: Efficiency of adenoviral expressions of these molecules in pig islets was approximately 80% at a multiplicity of infection of 100. In an in vitro assay, approximately 80% suppression of cytotoxicity was observed in membrane bound human FasL-expressing pig islets and 60% inhibition of CTL killing was displayed in decoy Fas expression pig islets. In an in vivo transplant model, prolonged survival of pig islets xenografts, expressing either membrane-bound human FasL or human decoy Fas genes, was elicited in comparison with that of control islets xenografts. CONCLUSION: The extracellular remodeling of either death receptor or death ligand genes by adenoviral expression was effective for the prevention of CTL-mediated xenocytotoxicity in pig islets. PMID- 19134164 TI - Xenotransplantation literature update May-August, 2008. PMID- 19134166 TI - High-resolution mapping of plasmid transcriptomes in different host bacteria. AB - BACKGROUND: Plasmids are extrachromosomal elements that replicate autonomously, and many can be transmitted between bacterial cells through conjugation. Although the transcription pattern of genes on a plasmid can be altered by a change in host background, the expression range of plasmid genes that will result in phenotypic variation has not been quantitatively investigated. RESULTS: Using a microarray with evenly tiled probes at a density of 9 bp, we mapped and quantified the transcripts of the carbazole catabolic plasmid pCAR1 in its original host Pseudomonas resinovorans CA10 and the transconjugant P. putida KT2440(pCAR1) during growth on either carbazole or succinate as the sole carbon source. We identified the operons in pCAR1, which consisted of nearly identical transcription units despite the difference in host background during growth on the same carbon source. In accordance with previous studies, the catabolic operons for carbazole degradation were upregulated during growth on carbazole in both hosts. However, our tiling array results also showed that several operons flanking the transfer gene cluster were transcribed at significantly higher levels in the transconjugant than in the original host. The number of transcripts and the positions of the transcription start sites agreed with our quantitative RT-PCR and primer extension results. CONCLUSION: Our tiling array results indicate that the levels of transcription for the operons on a plasmid can vary by host background. High-resolution mapping using an unbiased tiling array is a valuable tool for the simultaneous identification and quantification of prokaryotic transcriptomes including polycistronic operons and non-coding RNAs. PMID- 19134167 TI - Blood lactate levels in 31 female dogs with pyometra. AB - BACKGROUND: Canine pyometra is a life-threatening disease common in countries where spaying of dogs is not routinely performed. The disease is associated with endotoxemia, sepsis, systemic inflammatory response syndrome (SIRS) and a 3-4% mortality rate. Blood lactate analysis is clinically valuable in predicting prognosis and survival, evaluating tissue perfusion and treatment response in human and veterinary critical care settings. The aims of the present study were to investigate 1) the blood lactate levels of female dogs with pyometra by a hand held analyser and 2) if these levels are related with the clinical status or other biochemical or hematological disorders. METHODS: In total 31 female dogs with pyometra admitted for surgical ovariohysterectomy and 16 healthy female control dogs were included in the present study. A complete physical examination including SIRS-status determination was performed. Blood samples for lactate concentrations, hematological and biochemical parameters, acid-base and blood gas analysis and other laboratory parameters were collected and subsequently analysed. The diagnosis pyometra was verified with histopathological examination of the uterus and ovaries. Increased hospitalisation length and presence of SIRS were used as indicators of outcome. RESULTS: In the pyometra group the median blood lactate level was 1.6 mmol l(-1) (range <0.8-2.7 mmol l(-1)). In the control group the median lactate level was 1.2 mmol l(-1) (range <0.8-2.1 mmol l( 1)). Of the 31 bitches 19 (61%) fulfilled 2 or more criteria for SIRS at inclusion, 10 bitches (32%) fulfilled 3 of the SIRS criteria whereas none accomplished more than 3 criteria. Lactate levels did not differ significantly between the pyometra and control group, or between the SIRS positive and SIRS negative dogs with pyometra. Increased lactate concentration (>2.5 mmol l(-1)) was demonstrated in one female dog with pyometra (3%), and was not associated with longer hospitalisation or presence of SIRS. Lactate measurement was not indicative of peritonitis. None of the bitches died during or within two months of the hospital stay. The measurements of temperature, heart rate, respiratory rate, percentage bandforms of neutrophilic granulocytes, alpha2-globulins, creatinin, pvCO2, TCO2 and base excess showed significant differences between the SIRS positive and the SIRS negative pyometra cases. CONCLUSION: Increased blood lactate concentrations were demonstrated in 3% (1/31), and SIRS was present in 61% (19/31) of the female dogs with pyometra. Preoperative lactate levels were not related with presence of SIRS or prolonged hospitalisation. Lactate measurement was not indicative of peritonitis. The value of a single and repeated lactate analysis in more severely affected cases remains to be determined. PMID- 19134168 TI - Prevalence of latent tuberculosis infection among health care workers in a hospital for pulmonary diseases. AB - BACKGROUND: Little is known about the prevalence of latent tuberculosis infections (LTBI) in health care workers (HCW) in low-incidence countries especially in hospitals for pulmonary diseases. With Interferon-gamma release assays (IGRA), a new method for diagnosis of LTBI is available which is more specific than the tuberculin skin test (TST). OBJECTIVES: The study was designed to estimate prevalence of LTBI among 270 HCW in a Hospital of Pulmonary Diseases routinely screened for TB. METHODS: LTBI was assessed by the QuantiFERON-Gold In Tube (QFT-IT). Information on gender, age, workplace, job title, BCG vaccination and history of both TB and TST were collected using a standardised questionnaire. Adjusted odds ratios for potential risk factors for LTBI were calculated. RESULTS: The prevalence of LTBI was 7.2%. In HCW younger than 30 years LTBI prevalence was 3.5% and in those older than 50 years 22%. Physicians and nurses showed a higher prevalence rate than other professions (10.8% to 4.5%). The putative risk factors for LTBI were age (>50 year OR 9.3, 95%CI 2.5-33.7), working as physicians/nurses (OR 3. 95%CI 1.2-10.4) and no previous TST in medical history (OR 4.4, 95%CI 1.01-18.9) when compared to those with a negative TST. CONCLUSION: Prevalence of LTBI assessed by QFT-IT is low, this indicates a low infection risk even in hospitals for pulmonary diseases. No statement can be made regarding the occupational risk as compared to the general population because there are no LTBI prevalence data from Germany available. The higher LTBI prevalence rate in older HCWs might be due to the cohort effect or the longer time at risk. PMID- 19134169 TI - Linkage mapping of putative regulator genes of barley grain development characterized by expression profiling. AB - BACKGROUND: Barley (Hordeum vulgare L.) seed development is a highly regulated process with fine-tuned interaction of various tissues controlling distinct physiological events during prestorage, storage and dessication phase. As potential regulators involved within this process we studied 172 transcription factors and 204 kinases for their expression behaviour and anchored a subset of them to the barley linkage map to promote marker-assisted studies on barley grains. RESULTS: By a hierachical clustering of the expression profiles of 376 potential regulatory genes expressed in 37 different tissues, we found 50 regulators preferentially expressed in one of the three grain tissue fractions pericarp, endosperm and embryo during seed development. In addition, 27 regulators found to be expressed during both seed development and germination and 32 additional regulators are characteristically expressed in multiple tissues undergoing cell differentiation events during barley plant ontogeny. Another 96 regulators were, beside in the developing seed, ubiquitously expressed among all tissues of germinating seedlings as well as in reproductive tissues. SNP-marker development for those regulators resulted in anchoring 61 markers on the genetic linkage map of barley and the chromosomal assignment of another 12 loci by using wheat-barley addition lines. The SNP frequency ranged from 0.5 to 1.0 SNP/kb in the parents of the various mapping populations and was 2.3 SNP/kb over all eight lines tested. Exploration of macrosynteny to rice revealed that the chromosomal orders of the mapped putative regulatory factors were predominantly conserved during evolution. CONCLUSION: We identified expression patterns of major transcription factors and signaling related genes expressed during barley ontogeny and further assigned possible functions based on likely orthologs functionally well characterized in model plant species. The combined linkage map and reference expression map of regulators defined in the present study offers the possibility of further directed research of the functional role of regulators during seed development in barley. PMID- 19134170 TI - Acupuncture, or non-directive counselling versus usual care for the treatment of depression: a pilot study. AB - BACKGROUND: Depression is one of the most common reasons for consulting in primary care. Acupuncture is a popular complementary therapy choice for depression but its evidence base is poor with more robust high quality trials being required. More than half of depressed patients experience painful symptoms, with severe pain being associated with poor response to antidepressants. Acupuncture may have much to offer as an intervention for depression that also helps alleviate pain. Non-directive counselling is the most widely used psychological approach for depression in NHS settings, and provides a useful pragmatic comparison for acupuncture that would, according to our pre-trial qualitative research, be of high interest to doctors and patients. METHODS AND DESIGN: The pilot study uses five arms and involves a pragmatic design. All patients will continue to receive usual care. Four groups of patients will be allocated to acupuncture, or non-directive counselling, in addition to usual GP care. The acupuncture and counselling arms will be further split into two groups to explore different treatment regimens. The primary outcome measure is the BDI II. Potentially eligible patients will be screened for depression using the PHQ 9, which is also a secondary outcome measure. Other secondary measures include the SF 36 bodily pain subscale, the CORE OM, the WBQ-12 and the EQ5D. Health economic data will be collected and measures of therapeutic engagement will be used to compare patient's views of therapists and GPs. The study will employ a fully randomised preference design with collection of data on patient preferences and prior expectations. DISCUSSION: This study has been implemented, and data are currently being analysed to inform the design of a full scale trial. Two practical operational issues that impacted on study implementation are discussed. Firstly, the challenge of recruiting depressed patients via GP consultation. Secondly, the problem of poor uptake and high attrition for counselling and acupuncture, which appeared to be associated with poor questionnaire return, and resulted in missing data. These problems may be relevant to other researchers working in the area of depression, or similar illnesses, where patients may lack motivation and energy to engage in research, or attend for treatment. TRIAL REGISTRATION: Current Controlled Trials (ISRCTN 59267538). PMID- 19134171 TI - Phone and e-mail counselling are effective for weight management in an overweight working population: a randomized controlled trial. AB - BACKGROUND: The work setting provides an opportunity to introduce overweight (i.e., Body Mass Index >or= 25 kg/m2) adults to a weight management programme, but new approaches are needed in this setting. The main purpose of this study was to investigate the effectiveness of lifestyle counselling by phone or e-mail on body weight, in an overweight working population. Secondary purposes were to establish effects on waist circumference and lifestyle behaviours, and to assess which communication method is the most effective. METHODS: A randomized controlled trial with three treatments: intervention materials with phone counselling (phone group); a web-based intervention with e-mail counselling (internet group); and usual care, i.e. lifestyle brochures (control group). The interventions used lifestyle modification and lasted a maximum of six months. Subjects were 1386 employees, recruited from seven companies (67% male; mean age 43 (SD 8.6) y; mean BMI 29.6 (SD 3.5) kg/m2). Body weight was measured by research personnel and by questionnaire. Secondary outcomes fat, fruit and vegetable intake, physical activity and waist circumference were assessed by questionnaire. Measurements were done at baseline and after six months. Missing body weight was multiply imputed. RESULTS: Body weight reduced 1.5 kg (95% CI 2.2;-0.8, p < 0.001) in the phone group and 0.6 kg (95% CI -1.3; -0.01, p = 0.045) in the internet group, compared with controls. In completers analyses, weight and waist circumference in the phone group were reduced with 1.6 kg (95% CI -2.2;-1.0, p < 0.001) and 1.9 cm (95% CI -2.7;-1.0, p < 0.001) respectively, fat intake decreased with 1 fatpoint (1 to 4 grams)/day (95% CI -1.7;-0.2, p = 0.01) and physical activity increased with 866 METminutes/week (95% CI 203;1530, p = 0.01), compared with controls. The internet intervention resulted in a weight loss of 1.1 kg (95% CI -1.7;-0.5, p < 0.001) and a reduction in waist circumference of 1.2 cm (95% CI -2.1;-0.4, p = 0.01), in comparison with usual care. The phone group appeared to have more and larger changes than the internet group, but comparisons revealed no significant differences. CONCLUSION: Lifestyle counselling by phone and e-mail is effective for weight management in overweight employees and shows potential for use in the work setting. TRIAL REGISTRATION: ISCRTN04265725. PMID- 19134172 TI - Insight into the molecular requirements for pathogenicity of Fusarium oxysporum f. sp. lycopersici through large-scale insertional mutagenesis. AB - BACKGROUND: Fusarium oxysporum f. sp. lycopersici is the causal agent of vascular wilt disease in tomato. In order to gain more insight into the molecular processes in F. oxysporum necessary for pathogenesis and to uncover the genes involved, we used Agrobacterium-mediated insertional mutagenesis to generate 10,290 transformants and screened the transformants for loss or reduction of pathogenicity. RESULTS: This led to the identification of 106 pathogenicity mutants. Southern analysis revealed that the average T-DNA insertion is 1.4 and that 66% of the mutants carry a single T-DNA. Using TAIL-PCR, chromosomal T-DNA flanking regions were isolated and 111 potential pathogenicity genes were identified. CONCLUSIONS: Functional categorization of the potential pathogenicity genes indicates that certain cellular processes, such as amino acid and lipid metabolism, cell wall remodeling, protein translocation and protein degradation, seem to be important for full pathogenicity of F. oxysporum. Several known pathogenicity genes were identified, such as those encoding chitin synthase V, developmental regulator FlbA and phosphomannose isomerase. In addition, complementation and gene knock-out experiments confirmed that a glycosylphosphatidylinositol-anchored protein, thought to be involved in cell wall integrity, a transcriptional regulator, a protein with unknown function and peroxisome biogenesis are required for full pathogenicity of F. oxysporum. PMID- 19134173 TI - Modulators of arginine metabolism support cancer immunosurveillance. AB - BACKGROUND: Tumor-associated accrual of myeloid derived suppressor cells (MDSC) in the blood, lymphoid organs and tumor tissues may lead to perturbation of the arginine metabolism and impairment of the endogenous antitumor immunity. The objective of this study was to evaluate whether accumulation of MDSC occurred in Th2 prone BALB/c and Th1 biased C57BL/6 mice bearing the C26GM colon carcinoma and RMA T lymphoma, respectively, and to investigate whether N(G) nitro-L arginine methyl ester (L-NAME) and sildenafil, both modulators of the arginine metabolism, restored antitumor immunity. RESULTS: We report here that MDSC accumulate in the spleen and blood of mice irrespective of the mouse and tumor model used. Treatment of tumor-bearing mice with either the phosphodiesterase-5 inhibitor sildenafil or the nitric-oxide synthase (NOS) inhibitor L-NAME significantly restrained tumor growth and expanded the tumor-specific immune response. CONCLUSION: Our data emphasize the role of MDSC in modulating the endogenous tumor-specific immune response and underline the anti-neoplastic therapeutic potential of arginine metabolism modulators. PMID- 19134174 TI - Estimation of tumor heterogeneity using CGH array data. AB - BACKGROUND: Array-based comparative genomic hybridization (CGH) is a commonly used approach to detect DNA copy number variation in whole genome-wide screens. Several statistical methods have been proposed to define genomic segments with different copy numbers in cancer tumors. However, most tumors are heterogeneous and show variation in DNA copy numbers across tumor cells. The challenge is to reveal the copy number profiles of the subpopulations in a tumor and to estimate the percentage of each subpopulation. RESULTS: We describe a relation between experimental data and exact DNA copy number and develop a statistical method to reveal the heterogeneity of tumors containing a mixture of different-stage cells. Furthermore, we validate the method on simulated data and apply the method to 29 pairs of breast primary tumors and their matched lymph node metastases. CONCLUSION: We demonstrate a new method for CGH array analysis that allows a tumor sample to be classified according to its heterogeneity. The method gives an interpretable series of copy number profiles, one for each major subpopulation in a tumor. The profiles facilitate identification of copy number alterations in cancer development. PMID- 19134175 TI - Parasite resistance and the adaptive significance of sleep. AB - BACKGROUND: Sleep is a biological enigma. Despite occupying much of an animal's life, and having been scrutinized by numerous experimental studies, there is still no consensus on its function. Similarly, no hypothesis has yet explained why species have evolved such marked variation in their sleep requirements (from 3 to 20 hours a day in mammals). One intriguing but untested idea is that sleep has evolved by playing an important role in protecting animals from parasitic infection. This theory stems, in part, from clinical observations of intimate physiological links between sleep and the immune system. Here, we test this hypothesis by conducting comparative analyses of mammalian sleep, immune system parameters, and parasitism. RESULTS: We found that evolutionary increases in mammalian sleep durations are strongly associated with an enhancement of immune defences as measured by the number of immune cells circulating in peripheral blood. This appeared to be a generalized relationship that could be independently detected in 4 of the 5 immune cell types and in both of the main sleep phases. Importantly, no comparable relationships occur in related physiological systems that do not serve an immune function. Consistent with an influence of sleep on immune investment, mammalian species that sleep for longer periods also had substantially reduced levels of parasitic infection. CONCLUSION: These relationships suggest that parasite resistance has played an important role in the evolution of mammalian sleep. Species that have evolved longer sleep durations appear to be able to increase investment in their immune systems and be better protected from parasites. These results are neither predicted nor explained by conventional theories of sleep evolution, and suggest that sleep has a much wider role in disease resistance than is currently appreciated. PMID- 19134176 TI - Sleeping to fuel the immune system: mammalian sleep and resistance to parasites. AB - Sleep is an enigma. Why animals forgo eating and reproducing, while potentially increasing their risk of predation remains unknown. Although some may question whether all animals sleep, it is clear that all living organisms possess defenses against attack by pathogens. Immune responses of humans and animals are impaired by sleep loss, and responses to immune challenge include altered sleep. Thus, sleep is hypothesized to be a component of the acute phase response to infection and to function in host defense. Examining phylogenetic relationships among sleep parameters, components of the mammalian immune system and resistance to infection may provide insight into the evolution of sleep and lead to a greater appreciation for the role of sleep in host defense. PMID- 19134177 TI - Precision of field triage in patients brought to a trauma centre after introducing trauma team activation guidelines. AB - BACKGROUND: Field triage is important for regional trauma systems providing high sensitivity to avoid that severely injured are deprived access to trauma team resuscitation (undertriage), yet high specificity to avoid resource over utilization (overtriage). Previous informal trauma team activation (TTA) at Ulleval University Hospital (UUH) caused imprecise triage. We have analyzed triage precision after introduction of TTA guidelines. METHODS: Retrospective analysis of 7 years (2001-07) of prospectively collected trauma registry data for all patients with TTA or severe injury, defined as at least one of the following: Injury Severity Score (ISS) > 15, proximal penetrating injury, admitted ICU > 2 days, transferred intubated to another hospital within 2 days, dead from trauma within 30 days. Interhospital transfers to UUH and patients admitted by non healthcare personnel were excluded. Overtriage is the fraction of TTA where patients are not severely injured (1-positive predictive value); undertriage is the fraction of severely injured admitted without TTA (1-sensitivity). RESULTS: Of the 4,659 patients included in the study, 2,221 (48%) were severely injured. TTA occurred 4,440 times, only 2,002 of which for severely injured (overtriage 55%). Overall undertriage was 10%. Mechanism of injury was TTA criterion in 1,508 cases (34%), of which only 392 were severely injured (overtriage 74%). Paramedic manned prehospital services provided 66% overtriage and 17% undertriage, anaesthetist-manned services 35% overtriage and 2% undertriage. Falls, high age and admittance by paramedics were significantly associated with undertriage. A Triage-Revised Trauma Score (RTS) < 12 in the emergency department reduced the risk for undertriage compared to RTS = 12 (normal value). Field RTS was documented by anaesthetists in 64% of the patients compared to 33% among paramedics.Patients subject to undertriage had an ISS-adjusted Odds Ratio for 30 day mortality of 2.34 (95% CI 1.6-3.4, p < 0.001) compared to those correctly triaged to TTA. CONCLUSION: Triage precision had not improved after TTA guideline introduction. Anaesthetists perform precise trauma triage, whereas paramedics have potential for improvement. Skewed mission profiles makes comparison of differences in triage precision difficult, but criteria or the use of them may contribute. Massive undertriage among paramedics is of grave concern as patients exposed to undertriage had increased risk of dying. PMID- 19134178 TI - High expression of 5-lipoxygenase in normal and malignant mantle zone B lymphocytes. AB - BACKGROUND: Human B lymphocytes can produce leukotriene B4 but the biological function of the 5-lipoxygenase (5-LO) pathway in B cells is unclear. In order to better understand and define the role of 5-LO in B cells, we investigated the expression of 5-LO mRNA and protein in subsets of B cells from human tonsils and different types of B cell lymphoma. RESULTS: Based on RT-PCR and western blot/immunohistochemical staining, with a polyclonal antibody raised against 5 LO, high expression of 5-LO was found in mantle zone B cells from tonsils. By contrast, only a weak expression of 5-LO was detected in germinal centre cells and no expression in plasma cells from tonsils. This pattern of 5-LO expression was preserved in malignant lymphoma with high expression in mantle B cell lymphoma (MCL) and weak or no expression in follicular lymphoma. Primary leukemized MCL, so called B-prolymphocytic leukaemia cells, and MCL cell lines also expressed 5-LO and readily produced LTB4 after activation. CONCLUSION: The present report demonstrates the expression of 5-LO mainly in normal and malignant mantle zone B cells while the expression is low or absent in germinal centre B cells and plasma cells, indicating a role of the 5-LO pathway in B cells before the cells finally differentiate to plasma cells. PMID- 19134179 TI - "I should live and finish it": a qualitative inquiry into Turkish women's menopause experience. AB - BACKGROUND: While bio-medically, menopause could be treated as an illness, from a psychosocial and cultural perspective it could be seen as a "natural" process without requiring medication unless severe symptoms are present.Our objective is to explore the perceptions of Turkish women regarding menopause and Hormone Therapy (HT) to provide health care workers with an insight into the needs and expectations of postmenopausal women. METHODS: A qualitative inquiry through semi structured, in-depth interviews was used to explore the study questions. We used a purposive sampling and included an equal number of participants who complained about the climacteric symptoms and those who visited the outpatient department for a problem other than climacteric symptoms but when asked declared that they had been experiencing climacteric symptoms. The interview questions focused on two areas; 1) knowledge, experiences, attitudes and beliefs about menopause and; 2) menopause-related experiences and ways to cope with menopause and perception of HT. RESULTS: Most of the participants defined menopause as a natural transition process that one should go through. Cleanliness, maturity, comfort of not having a period and positive changes in health behaviour were the concepts positively attributed to menopause, whereas hot flushes, getting old and difficulties in relationships were the negatives. Osteoporosis was an important concern for most of the participants. To deal with the symptoms, the non pharmacological options were mostly favoured. CONCLUSION: To our knowledge, this is the first qualitative study which focuses on Turkish women's menopausal experiences. Menopause was thought to be a natural process which was characterised by positive and negative features. Understanding these features and their implications in these women's lives may assist healthcare workers in helping their clients with menopause. PMID- 19134180 TI - Contralateral approach to a carotid bifurcation aneurysm in a case of multiple intracranial aneurysms: a case report. AB - BACKGROUND: Traditionally, surgery of the anterior circulation aneurysms of the cerebral vasculature is dictated by the site of the lesion, excluding such midline lesions as anterior communication artery aneurysms. Few reports address the issue of using a single craniotomy to obliterate multiple aneurysms located in both hemispheres. CASE PRESENTATION: A 51 year-old Caucasian right handed housewife lady (weight 61 kg, height 159 cm) presented with a headache of acute onset which proved to be caused by acute subarachnoid hemorrhage. Cerebral computed tomographic angiography revealed multiple aneurysms. The patient underwent a right pterional craniotomy to obliterate right middle cerebral, distal basilar and left carotid bifurcation aneurysms. The post-operative course was uneventful. CONCLUSION: Despite technical difficulties of approaching cerebral vasculature through a contralateral craniotomy, this policy is advised in selected cases in which the benefits of unilateral craniotomy outweigh the risks of brain retraction. PMID- 19134181 TI - Integration and acceleration of virtual microscopy as the key to successful implementation into the routine diagnostic process. AB - BACKGROUND: The virtual microscopy is widely accepted in Pathology for educational purposes and teleconsultation but is far from the routine use in surgical pathology due to the technical requirements and some limitations. A technical problem is the limited bandwidth of a usual network and the delayed transmission rate and presentation time on the screen. METHODS: In this study the process of secondary diagnostic was evaluated using the "T.Konsult Pathologie" service of the Professional Association of German Pathologists within the German breast cancer screening program. The characteristics of the access to the WSI (Whole Slide Images) have been analyzed to explore the possibilities of prefetching and caching to reduce the presentation and transfer time with the goal to increase user acceptance. The log files of the web server were analyzed to reconstruct the movements of the pathologist on the WSI and to create the observation path. Using a specialized tool the observation paths were extracted automatically from the log files. The attributes linearity, 3-point-linearity, changes per request, and number of consecutive requests were calculated to design, develop and evaluate different caching and prefetching strategies. RESULTS: The analysis of the observation paths showed that a complete accordance of two image requests is a very rare event. But more frequently a partial covering of two requested image areas can be found. In total 257 diagnostic paths from 131 WSI have been extracted and analysed. On average a diagnostic path consists of 16 image requests and takes 189 seconds between first and last image request. The mean linearity was 0,41 and the mean 3-point-linearity 0,85. Three different caching algorithms have been compared with respect to hit rate and additional image requests on the WSI server. Tests demonstrated that 95% of the diagnostic paths could be loaded without any deletion of entries in the cache (cache size 12,2 Megapixel). If the image parts are stored after JPEG compression this complies with less than 2 MB. DISCUSSION: WSI telepathology is a technology which offers the possibility to break the limitations of conventional static telepathology. The complete histological slide may be investigated instead of sets of images of lesions sampled by the presenting pathologist. The benefit is demonstrated by the high diagnostic security of 95% accordance between first and second diagnosis. PMID- 19134182 TI - MegaSNPHunter: a learning approach to detect disease predisposition SNPs and high level interactions in genome wide association study. AB - BACKGROUND: The interactions of multiple single nucleotide polymorphisms (SNPs) are highly hypothesized to affect an individual's susceptibility to complex diseases. Although many works have been done to identify and quantify the importance of multi-SNP interactions, few of them could handle the genome wide data due to the combinatorial explosive search space and the difficulty to statistically evaluate the high-order interactions given limited samples. RESULTS: Three comparative experiments are designed to evaluate the performance of MegaSNPHunter. The first experiment uses synthetic data generated on the basis of epistasis models. The second one uses a genome wide study on Parkinson disease (data acquired by using Illumina HumanHap300 SNP chips). The third one chooses the rheumatoid arthritis study from Wellcome Trust Case Control Consortium (WTCCC) using Affymetrix GeneChip 500K Mapping Array Set. MegaSNPHunter outperforms the best solution in this area and reports many potential interactions for the two real studies. CONCLUSION: The experimental results on both synthetic data and two real data sets demonstrate that our proposed approach outperforms the best solution that is currently available in handling large-scale SNP data both in terms of speed and in terms of detection of potential interactions that were not identified before. To our knowledge, MegaSNPHunter is the first approach that is capable of identifying the disease-associated SNP interactions from WTCCC studies and is promising for practical disease prognosis. PMID- 19134183 TI - The association of posttraumatic stress disorder and metabolic syndrome: a study of increased health risk in veterans. AB - BACKGROUND: There is accumulating evidence for a link between trauma exposure, posttraumatic stress disorder (PTSD) and diminished health status. To assess PTSD related biological burden, we measured biological factors that comprise metabolic syndrome, an important established predictor of morbidity and mortality, as a correlate of long-term health risk in PTSD. METHODS: We analyzed clinical data from 253 male and female veterans, corresponding to five factors linked to metabolic syndrome (systolic and diastolic blood pressure, waist-to-hip ratio and fasting measures of high-density lipoprotein (HDL) cholesterol, serum triglycerides and plasma glucose concentration). Clinical cut-offs were defined for each biological parameter based on recommendations from the World Health Organization and the National Cholesterol Education Program. Controlling for relevant variables including sociodemographic variables, alcohol/substance/nicotine use and depression, we examined the impact of PTSD on metabolic syndrome using a logistic regression model. RESULTS: Two-fifths (40%) of the sample met criteria for metabolic syndrome. Of those with PTSD (n = 139), 43% met criteria for metabolic syndrome. The model predicted metabolic syndrome well (-2 log likelihood = 316.650, chi-squared = 23.731, p = 0.005). Veterans with higher severity of PTSD were more likely to meet diagnostic criteria for metabolic syndrome (Wald = 4.76, p = 0.03). CONCLUSION: These findings provide preliminary evidence linking higher severity of PTSD with risk factors for diminished health and increased morbidity, as represented by metabolic syndrome. PMID- 19134184 TI - Research priorities for non-pharmacological therapies for common musculoskeletal problems: nationally and internationally agreed recommendations. AB - BACKGROUND: Musculoskeletal problems such as low back pain, neck, knee and shoulder pain are leading causes of disability and activity limitation in adults and are most frequently managed within primary care. There is a clear trend towards large, high quality trials testing the effectiveness of common non pharmacological interventions for these conditions showing, at best, small to moderate benefits. This paper summarises the main lessons learnt from recent trials of the effectiveness of non-pharmacological therapies for common musculoskeletal conditions in primary care and provides agreed research priorities for future clinical trials. METHODS: Consensus development using nominal group techniques through national (UK) and international workshops. During a national Clinical Trials Thinktank workshop in April 2007 in the UK, a group of 30 senior researchers experienced in clinical trials for musculoskeletal conditions and 2 patient representatives debated the possible explanations for the findings of recent high quality trials of non-pharmacological interventions. Using the qualitative method of nominal group technique, these experts developed and ranked a set of priorities for future research, guided by the evidence from recent trials of treatments for common musculoskeletal problems. The recommendations from the national workshop were presented and further ranked at an international symposium (hosted in Canada) in June 2007. RESULTS: 22 recommended research priorities were developed, of which 12 reached consensus as priorities for future research from the UK workshop. The 12 recommendations were reduced to 7 agreed priorities at the international symposium. These were: to increase the focus on implementation (research into practice); to develop national musculoskeletal research networks in which large trials can be sited and smaller trials supported; to use more innovative trial designs such as those based on stepped care and subgrouping for targeted treatment models; to routinely incorporate health economic analysis into future trials; to include more patient centred outcome measures; to develop a core set of outcomes for new trials of interventions for musculoskeletal problems; and to focus on studies that advance methodological approaches for clinical trials in this field. CONCLUSION: A set of research priorities for future trials of non-pharmacological therapies for common musculoskeletal conditions has been developed and agreed through national (UK) and international consensus processes. These priorities provide useful direction for researchers and research funders alike and impetus for improvement in the quality and methodology of clinical trials in this field. PMID- 19134185 TI - Pdx-1 or Pdx-1-VP16 protein transduction induces beta-cell gene expression in liver-stem WB cells. AB - BACKGROUND: Pancreatic duodenal homeobox-1 (Pdx-1) or Pdx-1-VP16 gene transfer has been shown to induce in vitro rat liver-stem WB cell conversion into pancreatic endocrine precursor cells. High glucose conditions were necessary for further differentiation into functional insulin-producing cells. Pdx-1 has the ability to permeate different cell types due to an inherent protein transduction domain (PTD). In this study, we evaluated liver-to-pancreas conversion of WB cells following Pdx-1 or Pdx-1-VP16 protein transduction. FINDINGS: WB cells were grown in high glucose medium containing Pdx-1 or Pdx-1-VP16 recombinant proteins for two weeks. beta-like cell commitment was analysed by RT-PCR of pancreatic endocrine genes. We found that WB cells in high glucose culture spontaneously express pancreatic endocrine genes (Pdx-1, Ngn3, Nkx2.2, Kir6.2). Their further differentiation into beta-like cells expressing genes related to endocrine pancreas development (Ngn3, NeuroD, Pax4, Nkx2.2, Nkx6.1, Pdx-1) and beta-cell function (Glut-2, Kir6.2, insulin) was achieved only in the presence of Pdx-1( VP16) protein. CONCLUSION: These results demonstrate that Pdx-1(-VP16) protein transduction is instrumental for in vitro liver-to-pancreas conversion and is an alternative to gene therapy for beta-cell engineering for diabetes cell therapy. PMID- 19134186 TI - Intention-to-treat. What is the question? AB - It has become commonplace for Randomized Controlled Trials (RCTs) to be analyzed according to Intention-to-Treat (ITT) principles in which data from all subjects are used regardless of the subjects' adherence to protocol. While ITT analyses can provide useful information in some cases, they do not answer the question that motivates many RCTs, namely, whether the treatments differ in efficacy. ITT tends to reduce information by combining two questions, whether the intervention is effective and whether, as implemented, it has good compliance. Because these questions may be separate there is a risk of misuse. Two examples are presented that demonstrate this potential for abuse: a study on the effectiveness of vitamin E in reducing cardiovascular risk and comparisons of low fat and low carbohydrate diets. In the first case, a treatment that is demonstrably effective is described as without merit. In the second, ITT describes as the same, two diets that actually have different outcomes. These misuses of ITT are not atypical and are not technical problems in statistics but have real consequences for scientific principles and health recommendations. ITT analyses may answer the question of what happens when treatments are recommended but are inappropriate where separate information on adherence and performance is available. It is proposed that results of RCTs, or any experimental study, be reported, not in terms of the analyses that were performed, but rather in terms of the questions that the analyses can answer properly. PMID- 19134187 TI - Need to optimise infant feeding counselling: a cross-sectional survey among HIV positive mothers in Eastern Uganda. AB - BACKGROUND: The choice of infant feeding method is important for HIV-positive mothers in order to optimise the chance of survival of their infants and to minimise the risk of HIV transmission. The aim of this study was to investigate feeding practices, including breastfeeding, in the context of PMTCT for infants and children under two years of age born to HIV-positive mothers in Uganda. METHODS: In collaboration with The Aids Support Organisation Mbale, we conducted a cross-sectional survey involving 235 HIV-positive mothers in Uganda. Infant feeding practices, reasons for stopping breastfeeding, and breast health problems were studied. Breastfeeding duration was analysed using the Kaplan-Meier method based on retrospective recall. RESULTS: Breastfeeding was initiated by most of the mothers, but 20 of them (8.5%) opted exclusively for replacement feeding. Pre lacteal feeding was given to 150 (64%) infants and 65 (28%) practised exclusive breastfeeding during the first three days. One-fifth of the infants less than 6 months old were exclusively breastfed, the majority being complementary fed including breast milk. The median duration of breastfeeding was 12 months (95% confidence interval [CI] 11.5 to 12.5). Adjusted Cox regression analysis indicated that a mother's education, socio-economic status, participation in the PMTCT-program and her positive attitude to breastfeeding exclusively, were all associated with a reduction in breastfeeding duration. Median duration was 3 months (95% CI 0-10.2) among the most educated mothers, and 18 months (95% CI 15.0-21.0) among uneducated mothers. Participation in the PMTCT program and being socio-economically better-off were also associated with earlier cessation of breastfeeding (9 months [95% CI 7.2-10.8] vs. 14 months [95% CI 10.8-17.2] and 8 months [95% CI 5.9-10.1] vs. 17 months [95% CI 15.2-18.8], respectively). The main reasons for stopping breastfeeding were reported as: advice from health workers, maternal illness, and the HIV-positive status of the mother. CONCLUSION: Exclusive breastfeeding was uncommon. Exclusive replacement feeding was practised by few HIV-positive mothers. Well-educated mothers, mothers who were socio economically better-off and PMTCT-attendees had the shortest durations of breastfeeding. Further efforts are needed to optimise infant feeding counselling and to increase the feasibility of the recommendations. PMID- 19134188 TI - Effects and feasibility of a preventive intervention in sub-threshold and mild panic disorder: Results of a pilot study. AB - BACKGROUND: Panic disorder (PD) is a serious DSM-IV axis I disorder affecting up to 3% of the adult population each year. It is associated with a large burden of disease and extensive economic costs. This study aims to examine the effects and feasibility of the 'Don't Panic' course, a preventive cognitive behavioural intervention in sub-threshold and mild PD. It also compares the effectiveness of two modifications of the course (8 vs. 12 sessions). METHODS: The method used was a quasi-experimental two-group pre-post design with a baseline measurement (T0) and two follow-up measurements. Follow-ups were at the end of the intervention (T1) and six months later (T2). Primary outcome measure was the Panic Disorder Severity Scale-Self Report. A total of 114 participants suffering from panic attacks (mean age 42 years; 78% female) entered the study. RESULTS: The course participants showed a significant effect on the outcome measures at follow-up. Large effect sizes were found on panic symptoms, on symptoms of agoraphobia and on mental health-related quality of life at T1 and T2. Overall, the course leaders and the participants evaluated the course positively. There were no significant differences in outcome measures between the short and the long version of the course. CONCLUSION: The study suggests that people with sub threshold PD and mild PD could benefit from this preventive intervention and that the intervention might be feasible. Furthermore, the short version could be as effective as the long version. PMID- 19134189 TI - Synthesis of 4-pyridone-3-sulfate and an improved synthesis of 3-hydroxy-4 pyridone. AB - An improved synthesis of 3-hydroxy-4-pyridone via an Elbs oxidation of 4-pyridone and isolation of 4-pyridone-3-sulfate is described. PMID- 19134190 TI - Complement consumption in children with Plasmodium falciparum malaria. AB - BACKGROUND: Complement (C) can be activated during malaria, C components consumed and inflammatory mediators produced. This has potential to impair host innate defence. METHODS: In a case-control study, C activation was assessed by measuring serum haemolytic activity (CH50), functional activity of each pathway and levels of C3a, C4a and C5a in children presenting at Kisumu District Hospital, western Kenya, with severe malarial anaemia (SMA) or uncomplicated malaria (UM). RESULTS: CH50 median titers for lysis of sensitized sheep erythrocytes in SMA (8.6 U/mL) were below normal (34-70 U/mL) and were one-fourth the level in UM (34.6 U/mL (P < 0.001). Plasma C3a median levels were 10 times higher than in normals forSMA (3,200 ng/ml) and for UM (3,500 ng/ml), indicating substantial C activation in both groups. Similar trends were obtained for C4a and C5a. The activities of all three C pathways were greatly reduced in SMA compared to UM (9.9% vs 83.4% for CP, 0.09% vs 30.7% for MBL and 36.8% vs 87.7% for AP respectively, P < 0.001). CONCLUSION: These results indicate that, while C activation occurs in both SMA and UM, C consumption is excessive in SMA. It is speculated that in SMA, consumption of C exceeds its regeneration. PMID- 19134191 TI - Cognitive impairment and preferences for current health. AB - BACKGROUND: We assessed preferences for current health using the visual analogue scale (VAS), standard gamble (SG), time trade-off (TTO), and willingness to pay (WTP) in patients with cerebral aneurysms, a population vulnerable to cognitive deficits related to aneurysm bleeding or treatment. METHODS: We measured VAS, SG, TTO, and WTP values for current health in 165 outpatients with cerebral aneurysms. We assessed cognitive impairment with the Mini Mental State Examination (MMSE; scores < 24 = cognitive impairment). We examined the distributions of preference responses stratified by cognitive status, and the relationship between preferences and cognitive impairment, patient characteristics, and aneurysm history. RESULTS: Eleven patients (7%) had MMSE scores < 24. The distribution of preferences responses from patients with cognitive impairment had greater variance (SG, 0.39 vs. 0.21, P = 0.001; TTO, 0.36 vs. 0.24, P = 0.017) and altered morphology (VAS, P = 0.012; SG, P = 0.023) compared to the responses of unimpaired patients. There was good correlation between most preference measures for unimpaired patients (VAS:TTO, rho = 0.19, P = 0.018; SG:TTO, rho = 0.36, P < 0.001; SG:WTP, rho = -0.33, P < 0.001) and a trend towards significance with another pairing (VAS:WTP, rho = 0.16, P = 0.054). In subjects with cognitive impairment, there was a significant correlation only between VAS and TTO scores (rho = 0.76, P = 0.023). Separate regression models showed that cognitive impairment was associated with lower preferences on the VAS (beta = -0.12, P = 0.048), SG (beta = -0.23, P = 0.002), and TTO (beta = -0.17, P = 0.035). CONCLUSION: Cognitive impairment is associated with lower preferences for current health in patients with cerebral aneurysms. Cognitively impaired patients have poor inter-preference test correlations and different response distributions compared to unimpaired patients. PMID- 19134192 TI - BODE index versus GOLD classification for explaining anxious and depressive symptoms in patients with COPD - a cross-sectional study. AB - BACKGROUND: Anxiety and depression are common and treatable risk factors for re hospitalisation and death in patients with COPD. The degree of lung function impairment does not sufficiently explain anxiety and depression. The BODE index allows a functional classification of COPD beyond FEV1. The aim of this cross sectional study was (1) to test whether the BODE index is superior to the GOLD classification for explaining anxious and depressive symptoms; and (2) to assess which components of the BODE index are associated with these psychological aspects of COPD. METHODS: COPD was classified according to the GOLD stages based on FEV(1%predicted) in 122 stable patients with COPD. An additional four stage classification was constructed based on the quartiles of the BODE index. The hospital anxiety and depression scale was used to assess anxious and depressive symptoms. RESULTS: The overall prevalence of anxious and depressive symptoms was 49% and 52%, respectively. The prevalence of anxious symptoms increased with increasing BODE stages but not with increasing GOLD stages. The prevalence of depressive symptoms increased with both increasing GOLD and BODE stages. The BODE index was superior to FEV(1%predicted) for explaining anxious and depressive symptoms. Anxious symptoms were explained by dyspnoea. Depressive symptoms were explained by both dyspnoea and reduced exercise capacity. CONCLUSION: The BODE index is superior to the GOLD classification for explaining anxious and depressive symptoms in COPD patients. These psychological consequences of the disease may play a role in future classification systems of COPD. PMID- 19134193 TI - Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries. AB - BACKGROUND: Atherosclerosis affects aorta, coronary, carotid, and iliac arteries most frequently than any other body vessel. There may be common molecular pathways sustaining this process. Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis. RESULTS: We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression. We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis. Caveolae and JAK/STAT pathways, and S100A9/S100A8 interacting proteins are certainly involved in the development of vascular disease. We found that the system of caveolae is directly connected with genes that respond to hormone receptors, and indirectly with the apoptosis pathway. Cytokines, chemokines and growth factors released in the blood flux were investigated in parallel. High levels of RANTES, IL-1ra, MIP 1 alpha, MIP-1 beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-17, PDGF-BB, VEGF and IFN gamma were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels. CONCLUSION: The pattern of cytokine and S100A9/S100A8 up regulation characterizes atherosclerosis as a proinflammatory disorder. Activation of the JAK/STAT pathway is confirmed by the up-regulation of IL-6, STAT1, ISGF3G and IL10RA genes in coronary and carotid plaques. The functional network constructed in our research is an evidence of the central role of STAT protein and the caveolae system to contribute to preserve the plaque. Moreover, Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype. PMID- 19134194 TI - Tissue microarray analysis of eIF4E and its downstream effector proteins in human breast cancer. AB - BACKGROUND: Eukaryotic initiation factor 4E (eIF4E) is elevated in many cancers and is a prognostic indicator in breast cancer. Many pro-tumorigenic proteins are selectively translated via eIF4E, including c-Myc, cyclin D1, ornithine decarboxylase (ODC), vascular endothelial growth factor (VEGF) and Tousled-like kinase 1B (TLK1B). However, western blot analysis of these factors in human breast cancer has been limited by the availability of fresh frozen tissue and the labor-intensive nature of the multiple assays required. Our goal was to validate whether formalin-fixed, paraffin-embedded tissues arranged in a tissue microarray (TMA) format would be more efficient than the use of fresh-frozen tissue and western blot to test multiple downstream gene products. RESULTS: Breast tumor TMAs were stained immunohistochemically and quantitated using the ARIOL imaging system. In the TMAs, eIF4E levels correlated strongly with c-Myc, cyclin D1, TLK1B, VEGF, and ODC. Western blot comparisons of eIF4E vs. TLK1B were consistent with the immunohistochemical results. Consistent with our previous western blot results, eIF4E did not correlate with node status, ER, PR, or HER-2/neu. CONCLUSION: We conclude that the TMA technique yields similar results as the western blot technique and can be more efficient and thorough in the evaluation of several products downstream of eIF4E. PMID- 19134195 TI - Iron oxide nanoparticles induce human microvascular endothelial cell permeability through reactive oxygen species production and microtubule remodeling. AB - BACKGROUND: Engineered iron nanoparticles are being explored for the development of biomedical applications and many other industry purposes. However, to date little is known concerning the precise mechanisms of translocation of iron nanoparticles into targeted tissues and organs from blood circulation, as well as the underlying implications of potential harmful health effects in human. RESULTS: The confocal microscopy imaging analysis demonstrates that exposure to engineered iron nanoparticles induces an increase in cell permeability in human microvascular endothelial cells. Our studies further reveal iron nanoparticles enhance the permeability through the production of reactive oxygen species (ROS) and the stabilization of microtubules. We also showed Akt/GSK-3beta signaling pathways are involved in iron nanoparticle-induced cell permeability. The inhibition of ROS demonstrate ROS play a major role in regulating Akt/GSK-3beta - mediated cell permeability upon iron nanoparticle exposure. These results provide new insights into the bioreactivity of engineered iron nanoparticles which can inform potential applications in medical imaging or drug delivery. CONCLUSION: Our results indicate that exposure to iron nanoparticles induces an increase in endothelial cell permeability through ROS oxidative stress-modulated microtubule remodeling. The findings from this study provide new understandings on the effects of nanoparticles on vascular transport of macromolecules and drugs. PMID- 19134196 TI - Gene expression profiles during early differentiation of mouse embryonic stem cells. AB - BACKGROUND: Understanding the mechanisms controlling stem cell differentiation is the key to future advances in tissue and organ regeneration. Embryonic stem (ES) cell differentiation can be triggered by embryoid body (EB) formation, which involves ES cell aggregation in suspension. EB growth in the absence of leukaemia inhibitory factor (LIF) leads EBs to mimic early embryonic development, giving rise to markers representative of endoderm, mesoderm and ectoderm. Here, we have used microarrays to investigate differences in gene expression between 3 undifferentiated ES cell lines, and also between undifferentiated ES cells and Day 1-4 EBs. RESULTS: An initial array study identified 4 gene expression changes between 3 undifferentiated ES cell lines. Tissue culture conditions for ES differentiation were then optimized to give the maximum range of gene expression and growth. -Undifferentiated ES cells and EBs cultured with and without LIF at each day for 4 days were subjected to microarray analysis. -Differential expression of 23 genes was identified. 13 of these were also differentially regulated in a separate array comparison between undifferentiated ES cells and compartments of very early embryos. A high degree of inter-replicate variability was noted when confirming array results. Using a panel of marker genes, RNA amplification and RT-PCR, we examined expression pattern variation between individual -D4-Lif EBs. We found that individual EBs selected from the same dish were highly variable in gene expression profile. CONCLUSION: ES cell lines derived from different mouse strains and carrying different genetic modifications are almost invariant in gene expression profile under conditions used to maintain pluripotency. Tissue culture conditions that give the widest range of gene expression and maximise EB growth involve the use of 20% serum and starting cell numbers of 1000 per EB. 23 genes of importance to early development have been identified; more than half of these are also identified using similar studies, thus validating our results. EBs cultured in the same dish vary widely in terms of their gene expression (and hence, undoubtedly, in their future differentiation potential). This may explain some of the inherent variability in differentiation protocols that use EBs. PMID- 19134197 TI - Seasonal prevalence of malaria in West Sumba district, Indonesia. AB - BACKGROUND: Accurate information about the burden of malaria infection at the district or provincial level is required both to plan and assess local malaria control efforts. Although many studies of malaria epidemiology, immunology, and drug resistance have been conducted at many sites in Indonesia, there is little published literature describing malaria prevalence at the district, provincial, or national level. METHODS: Two stage cluster sampling malaria prevalence surveys were conducted in the wet season and dry season across West Sumba, Nusa Tenggara Province, Indonesia. RESULTS: Eight thousand eight hundred seventy samples were collected from 45 sub-villages in the surveys. The overall prevalence of malaria infection in the West Sumba District was 6.83% (95% CI, 4.40, 9.26) in the wet season and 4.95% (95% CI, 3.01, 6.90) in the dry. In the wet season Plasmodium falciparum accounted for 70% of infections; in the dry season P. falciparum and Plasmodium vivax were present in equal proportion. Malaria prevalence varied substantially across the district; prevalences in individual sub-villages ranged from 0-34%. The greatest malaria prevalence was in children and teenagers; the geometric mean parasitaemia in infected individuals decreased with age. Malaria infection was clearly associated with decreased haemoglobin concentration in children under 10 years of age, but it is not clear whether this association is causal. CONCLUSION: Malaria is hypoendemic to mesoendemic in West Sumba, Indonesia. The age distribution of parasitaemia suggests that transmission has been stable enough to induce some clinical immunity. These prevalence data will aid the design of future malaria control efforts and will serve as a baseline against which the results of current and future control efforts can be assessed. PMID- 19134198 TI - Characterization of the meningococcal DNA glycosylase Fpg involved in base excision repair. AB - BACKGROUND: Neisseria meningitidis, the causative agent of meningococcal disease, is exposed to high levels of reactive oxygen species inside its exclusive human host. The DNA glycosylase Fpg of the base excision repair pathway (BER) is a central player in the correction of oxidative DNA damage. This study aimed at characterizing the meningococcal Fpg and its role in DNA repair. RESULTS: The deduced N. meningitidis Fpg amino acid sequence was highly homologous to other Fpg orthologues, with particularly high conservation of functional domains. As for most N. meningitidis DNA repair genes, the fpg gene contained a DNA uptake sequence mediating efficient transformation of DNA. The recombinant N. meningitidis Fpg protein was over-expressed, purified to homogeneity and assessed for enzymatic activity. N. meningitidis Fpg was found to remove 2,6-diamino-4 hydroxy-5-formamidopyrimidine (faPy) lesions and 7,8-dihydro-8-oxo-2' deoxyguanosine (8oxoG) opposite of C, T and G and to a lesser extent opposite of A. Moreover, the N. meningitidis fpg single mutant was only slightly affected in terms of an increase in the frequency of phase variation as compared to a mismatch repair mutant. CONCLUSION: Collectively, these findings show that meningococcal Fpg functions are similar to those of prototype Fpg orthologues in other bacterial species. PMID- 19134199 TI - MBA: a literature mining system for extracting biomedical abbreviations. AB - BACKGROUND: The exploding growth of the biomedical literature presents many challenges for biological researchers. One such challenge is from the use of a great deal of abbreviations. Extracting abbreviations and their definitions accurately is very helpful to biologists and also facilitates biomedical text analysis. Existing approaches fall into four broad categories: rule based, machine learning based, text alignment based and statistically based. State of the art methods either focus exclusively on acronym-type abbreviations, or could not recognize rare abbreviations. We propose a systematic method to extract abbreviations effectively. At first a scoring method is used to classify the abbreviations into acronym-type and non-acronym-type abbreviations, and then their corresponding definitions are identified by two different methods: text alignment algorithm for the former, statistical method for the latter. RESULTS: A literature mining system MBA was constructed to extract both acronym-type and non acronym-type abbreviations. An abbreviation-tagged literature corpus, called Medstract gold standard corpus, was used to evaluate the system. MBA achieved a recall of 88% at the precision of 91% on the Medstract gold-standard EVALUATION Corpus. CONCLUSION: We present a new literature mining system MBA for extracting biomedical abbreviations. Our evaluation demonstrates that the MBA system performs better than the others. It can identify the definition of not only acronym-type abbreviations including a little irregular acronym-type abbreviations (e.g., ), but also non-acronym type abbreviations (e.g., ). PMID- 19134200 TI - No evidence of major effects in several Toll-like receptor gene polymorphisms in rheumatoid arthritis. AB - INTRODUCTION: The objective was to study the potential genetic contribution of Toll-like receptor (TLR) genes in rheumatoid arthritis (RA). TLRs bind to pathogen-associated molecular patterns, and TLR genes influence both proinflammatory cytokine production and autoimmune responses. Host-pathogen interactions are involved in RA physiopathology. METHODS: We tested SNPs of five TLR genes (TLR9, TLR2, TLR6, TLR1, and TLR4) in a cohort of 100 French families with RA. Genotypes were analyzed using the transmission disequilibrium test. As TLR2, TLR6, and TLR1 are located on chromosome 4, we determined the haplotype relative risk. Analyses were performed in subgroups defined by status for rheumatoid factor, anti-cyclic citrullinated peptide autoantibodies, and erosions. RESULTS: We found no disequilibrium in allele transmission for any of the SNPs of the five TLR genes. In subgroup analyses, no associations were detected linking TLR9, TLR2, or TLR9/TLR2 to rheumatoid factor, anti-cyclic citrullinated peptide autoantibodies, or erosions. Haplotype analysis of the polymorphisms showed no haplotype associations in any of the subgroups. CONCLUSIONS: We found no evidence of major effects of TLR gene polymorphisms in RA, although we tested different TLR phenotypes. Moreover, no associations were noted with autoantibody production or erosions. PMID- 19134201 TI - Sub-microscopic infections and long-term recrudescence of Plasmodium falciparum in Mozambican pregnant women. AB - BACKGROUND: Control of malaria in pregnancy remains a public health challenge. Improvements in its correct diagnosis and the adequacy of protocols to evaluate anti-malarial drug efficacy in pregnancy, are essential to achieve this goal. METHODS: The presence of Plasmodium falciparum was assessed by real-time (RT) PCR in 284 blood samples from pregnant women with clinical complaints suggestive of malaria, attending the maternity clinic of a Mozambican rural hospital. Parasite recrudescences in 33 consecutive paired episodes during the same pregnancy were identified by msp1 and msp2 genotyping. RESULTS: Prevalence of parasitaemia by microscopy was 5.3% (15/284) and 23.2% (66/284) by RT-PCR. Sensitivity of microscopy, compared to RT-PCR detection, was 22.7%. Risk of maternal anaemia was higher in PCR-positive women than in PCR-negative women (odds ratio [OR] = 1.92, 95% confidence interval [CI] 1.09-3.36). Genotyping confirmed that recrudescence after malaria treatment occurred in 7 (21%) out of 33 pregnant women with consecutive episodes during the same pregnancy (time range between recrudescent episodes: 14 to 187 days). CONCLUSION: More accurate and sensitive diagnostic indicators of malaria infection in pregnancy are needed to improve malaria control. Longer follow-up periods than the standard in vivo drug efficacy protocol should be used to assess anti-malarial drug efficacy in pregnancy. PMID- 19134202 TI - Use of task-shifting to rapidly scale-up HIV treatment services: experiences from Lusaka, Zambia. AB - The World Health Organization advocates task-shifting, the process of delegating clinical care functions from more specialized to less specialized health workers, as a strategy to achieve the United Nations Millennium Development Goals. However, there is a dearth of literature describing task shifting in sub-Saharan Africa, where services for antiretroviral therapy (ART) have scaled up rapidly in the face of generalized human resource crises. As part of ART services expansion in Lusaka, Zambia, we implemented a comprehensive task-shifting program among existing health providers and community-based workers. Training begins with didactic sessions targeting specialized skill sets. This is followed by an intensive period of practical mentorship, where providers are paired with trainers before working independently. We provide on-going quality assessment using key indicators of clinical care quality at each site. Program performance is reviewed with clinic-based staff quarterly. When problems are identified, clinic staff members design and implement specific interventions to address targeted areas. From 2005 to 2007, we trained 516 health providers in adult HIV treatment; 270 in pediatric HIV treatment; 341 in adherence counseling; 91 in a specialty nurse "triage" course, and 93 in an intensive clinical mentorship program. On-going quality assessment demonstrated improvement across clinical care quality indicators, despite rapidly growing patient volumes. Our task shifting strategy was designed to address current health care worker needs and to sustain ART scale-up activities. While this approach has been successful, long term solutions to the human resource crisis are also urgently needed to expand the number of providers and to slow staff migration out of the region. PMID- 19134203 TI - Structural factors associated with an increased risk of HIV and sexually transmitted infection transmission among street-involved youth. AB - BACKGROUND: The prevalence of HIV and sexually transmitted infections (STIs) among street-involved youth greatly exceed that of the general adolescent population; however, little is known regarding the structural factors that influence disease transmission risk among this population. METHODS: Between September 2005 and October 2006, 529 street-involved youth were enroled in a prospective cohort known as the At Risk Youth Study (ARYS). We examined structural factors associated with number of sex partners using quasi-Poisson regression and consistent condom use using logistic regression. RESULTS: At baseline, 415 (78.4%) were sexually active, of whom 253 (61.0%) reported multiple sex partners and 288 (69.4%) reported inconsistent condom use in the past six months. In multivariate analysis, self-reported barriers to health services were inversely associated with consistent condom use (adjusted odds ratio [aOR] = 0.52, 95%CI: 0.25 - 1.07). Structural factors that were associated with greater numbers of sex partners included homelessness (adjusted incidence rate ratio [aIRR] = 1.54, 95%CI: 1.11 - 2.14) and having an area restriction that affects access to services (aIRR = 2.32, 95%CI: 1.28 - 4.18). Being searched or detained by the police was significant for males (aIRR = 1.36, 95%CI: 1.02 - 1.81). CONCLUSION: Although limited by its cross-sectional design, our study found several structural factors amenable to policy-level interventions independently associated with sexual risk behaviours. These findings imply that the criminalization and displacement of street-involved youth may increase the likelihood that youth will engage in sexual risk behaviours and exacerbate the negative impact of resultant health outcomes. Moreover, our findings indicate that environmental-structural interventions may help to reduce the burden of these diseases among street youth in urban settings. PMID- 19134205 TI - Incidence and risk factors of exacerbations among COPD patients in primary health care: APMPOC study. AB - BACKGROUND: Worldwide, chronic obstructive pulmonary disease (COPD) is the fourth cause of death. Exacerbations have a negative impact on the prognosis of COPD and the frequency and severity of these episodes are associated with a higher patient mortality. Exacerbations are the first cause of decompensation, hospital admission and death in COPD. The incidence of exacerbations has mainly been estimated in populations of patients with moderate-severe COPD requiring hospital care. However, little is known regarding the epidemiology of exacerbations in patients with less severe COPD forms. It is therefore possible that a high number of these less severe forms of exacerbations are underdiagnosed and may, in the long-term, have certain prognostic importance for the COPD evolution. The aim of this study was to know the incidence and risk factors associated with exacerbations in patients with COPD in primary care. METHODS AND DESIGN: A prospective, observational, 3-phase, multicentre study will be performed involving: baseline evaluation, follow up and final evaluation. A total of 685 smokers or ex-smokers from 40 to 80 years of age with COPD, without acute respiratory disease or any other long-term respiratory disease will be randomly selected among the population assigned to 21 primary care centres. The diagnosis of COPD and its severity will be confirmed by spirometry. Information regarding the baseline situation, quality of life and exposure to contaminants or other factors potentially related to exacerbations will be collected. A group of 354 patients with confirmed COPD of varying severity will be followed for one year through monthly telephone calls and daily reporting of symptoms with the aim of detecting all the exacerbations which occur. These patients will be evaluated again at the end of the study and the incidence of exacerbations and associated relative risks will be estimated by negative binomial regression. DISCUSSION: The results will be relevant to provide knowledge about natural history of the initial phases of the COPD and the impact and incidence of the exacerbations on the patients with mild-moderate forms of the disease. These data may be important to know the milder forms of exacerbation which are often silent or very little expressed clinically. PMID- 19134206 TI - YKL-40 tissue expression and plasma levels in patients with ovarian cancer. AB - BACKGROUND: YKL-40 (chitinase-3-like-1) is a member of "mammalian chitinase-like proteins". The protein is expressed in many types of cancer cells and the highest plasma YKL-40 levels have been found in patients with metastatic disease, short recurrence/progression-free intervals, and short overall survival. The aim of the study was to determine the expression of YKL-40 in tumor tissue and plasma in patients with borderline ovarian tumor or epithelial ovarian cancer (OC), and investigate prognostic value of this marker. METHODS: YKL-40 protein expression was determined by immunohistochemistry in tissue arrays from 181 borderline tumors and 473 OC. Plasma YKL-40 was determined by ELISA in preoperative samples from 19 patients with borderline tumor and 76 OC patients. RESULTS: YKL-40 protein expression was found in cancer cells, tumor associated macrophages, neutrophils and mast cells. The tumor cell expression was higher in OC than in borderline tumors (p = 0.001), and associated with FIGO stage (p < 0.0001) and histological subtype (p = 0.0009). Positive YKL-40 expression (>or= 5% staining) was not associated with reduced survival. Plasma YKL-40 was also higher in patients with OC than in patients with borderline tumors (p < 0.0001), and it was positively correlated to serum CA-125 (p < 0.0001) and FIGO stage (p = 0.0001). Univariate Cox analysis of plasma YKL-40 showed association with overall survival (p < 0.0001). Multivariate Cox analysis, including plasma YKL-40, serum CA125, FIGO stage, age and radicality after primary surgery as variables, showed that elevated plasma YKL-40 was associated with a shorter survival (HR = 2.13, 95% CI: 1.40-3.25, p = 0.0004). CONCLUSION: YKL-40 in OC tissue and plasma are related to stage and histology, but only plasma YKL-40 is a prognostic biomarker in patients with OC. PMID- 19134207 TI - The prevalence of chronic diseases and major disease risk factors at different ages among 150,000 men and women living in Mexico City: cross-sectional analyses of a prospective study. AB - BACKGROUND: While most of the global burden from chronic diseases, and especially vascular diseases, is now borne by low and middle-income countries, few large scale epidemiological studies of chronic diseases in such countries have been performed. METHODS: From 1998-2004, 52 584 men and 106 962 women aged > or =35 years were visited in their homes in Mexico City. Self reported diagnoses of chronic diseases and major disease risk factors were ascertained and physical measurements taken. Age- and sex-specific prevalences and means were analysed. RESULTS: After about age 50 years, diabetes was extremely common - for example, 23.8% of men and 26.9% of women aged 65-74 reported a diagnosis. By comparison, ischaemic heart disease was reported by 4.8% of men and 3.0% of women aged 65-74, a history of stroke by 2.8% and 2.3%, respectively, and a history of cancer by 1.3% and 2.1%. Cancer history was generally more common among women than men - the excess being largest in middle-age, due to breast and cervical cancer. At older ages, the gap narrowed because of an increasing prevalence of prostate cancer. 51% of men and 25% of women aged 35-54 smoked cigarettes, while 29% of men and 41% of women aged 35-54 were obese (i.e. BMI > or =30 kg/m2). The prevalence of treated hypertension or measured blood pressure > or =140/90 mmHg increased about 50% more steeply with age among women than men, to 66% of women and 58% of men aged 65-74. Physical inactivity was highly prevalent but daily alcohol drinking was relatively uncommon. CONCLUSION: Diabetes, obesity and tobacco smoking are highly prevalent among adults living in Mexico City. Long term follow-up of this and other cohorts will establish the relevance of such factors to the major causes of death and disability in Mexico. PMID- 19134208 TI - RepPop: a database for repetitive elements in Populus trichocarpa. AB - BACKGROUND: Populus trichocarpa is the first tree genome to be completed, and its whole genome is currently being assembled. No functional annotation about the repetitive elements in the Populus trichocarpa genome is currently available. RESULTS: We predicted 9,623 repetitive elements in the Populus trichocarpa genome, and assigned functions to 3,075 of them (31.95%). The 9,623 repetitive elements cover approximately 40% of the current (partially) assembled genome. Among the 9,623 repetitive elements, 668 have copies only in the contigs that have not been assigned to one of the 19 chromosome while the rest all have copies in the partially assembled chromosomes. CONCLUSION: All the predicted data are organized into an easy-to-use web-browsable database, RepPop. Various search capabilities are provided against the RepPop database. A Wiki system has been set up to facilitate functional annotation and curation of the repetitive elements by a community rather than just the database developer. The database RepPop will facilitate the assembling and functional characterization of the Populus trichocarpa genome. PMID- 19134209 TI - Acquired nonobstructive urinary bladder diverticulum: a case report. AB - INTRODUCTION: Urinary bladder diverticula are frequently resulting from obstructions. Our literature review did not reveal any cases of acquired urinary bladder diverticulum caused by long-term transurethral catheterization. CASE PRESENTATION: We report a rare case of a nonobstructive big urinary bladder diverticulum developed after a long-term urethral catheterization in a 62-year old male diabetic patient with normal subvesical urinary tract. The diverticulum was demonstrated by ultrasonography. Its formation was associated with the decubital changes of the bladder wall when the Foley catheter stayed for a longer period. CONCLUSION: Ultrasonographic examination of the urinary bladder is necessary to exclude such complication after a long-lasting catheterization as well as to maximally restrict the catheter's stay in the urinary bladder. PMID- 19134210 TI - Immunome knowledge base (IKB): an integrated service for immunome research. AB - BACKGROUND: Functioning of the immune system requires the coordinated expression and action of many genes and proteins. With the emergence of high-throughput technologies, a great amount of molecular data is available for the genes and proteins of the immune system. However, these data are scattered into several databases and literature and therefore integration is needed. DESCRIPTION: The Immunome Knowledge Base (IKB) is a dedicated resource for immunological information. We identified and collected genes that are essential for the immunome. Nucleotide and protein sequences, as well as information about the related pseudogenes are available for 893 human essential immunome genes. To allow the study of the evolution of the immune system, data for the orthologs of human genes was collected. In addition to the human immunome, ortholog groups of 1811 metazoan immunity genes are available with information about the evidence of their immunity function. IKB combines three previous databases and several additional data items in an integrated system. CONCLUSION: IKB provides in one single service access to several databases and resources and contains plenty of new data about immune system. The most recent addition is variation data on genomic, transcriptomic and proteomic levels for all the immunome genes and proteins. In the future, more data will be added on the function of these genes. The service has a free and public web interface. PMID- 19134204 TI - Feeling manipulated: cytomegalovirus immune manipulation. AB - No one likes to feel like they have been manipulated, but in the case of cytomegalovirus (CMV) immune manipulation, we do not really have much choice. Whether you call it CMV immune modulation, manipulation, or evasion, the bottom line is that CMV alters the immune response in such a way to allow the establishment of latency with lifelong shedding. With millions of years of coevolution within their hosts, CMVs, like other herpesviruses, encode numerous proteins that can broadly influence the magnitude and quality of both innate and adaptive immune responses. These viral proteins include both homologues of host proteins, such as MHC class I or chemokine homologues, and proteins with little similarity to any other known proteins, such as the chemokine binding protein. Although a strong immune response is launched against CMV, these virally encoded proteins can interfere with the host's ability to efficiently recognize and clear virus, while others induce or alter specific immune responses to benefit viral replication or spread within the host. Modulation of host immunity allows survival of both the virus and the host. One way of describing it would be a kind of "mutually assured survival" (as opposed to MAD, Mutually Assured Destruction). Evaluation of this relationship provides important insights into the life cycle of CMV as well as a greater understanding of the complexity of the immune response to pathogens in general. PMID- 19134211 TI - The end of the era of generosity? Global health amid economic crisis. AB - In the past decade donor commitments to health have increased by 200 percent. Correspondingly, there has been a swell of new players in the global health landscape. The unprecedented, global response to a single disease, HIV/AIDS, has been responsible for a substantial portion of this boon. Numerous health success have followed this windfall of funding and attention, yet the food, fuel, and economic crises of 2008 have shown the vulnerabilities of health and development initiatives focused on short term wins and reliant on a constant flow of foreign funding. For too long, the international community has responded to global health and development challenges with emergency solutions that often reflect the donor's priorities, values, and political leanings, rather than funding durable health systems that can withstand crises. Progress towards achieving the Millennium Development Goals has stalled in many countries. Disease specific initiatives have weakened health systems and limited efforts to improve maternal and child health. As we enter this era of scarce resources, there is a need to return to the foundations of the Alma Ata Declaration signed thirty years ago with the goal of providing universal access to primary healthcare. The global health community must now objectively evaluate how we can most effectively respond to the crises of 2008 and take advantage of this moment of extraordinary attention for global health and translate it into long term, sustainable health improvements for all. PMID- 19134213 TI - Body aches, tender bones and rapid loss of weight: a case report. AB - INTRODUCTION: Bone metastases presenting with pain and body-ache may be the first presentation of carcinoma in about a fourth of patients with cancer. Radiologically majority of the metastases are osteolytic and multiple. Sometimes these may be confused with infective or inflammatory conditions, particularly in young individuals, and degenerative conditions of the spine and hip in elderly, which may delay the diagnosis and treatment leading to poor outcomes. CASE PRESENTATION: A 30 year old non-smoking male teetotaller presented with intermittent, high-grade nocturnal fever with night sweats of one year. He also had low back ache over his right hip. We found him febrile, pale and his long bones, ribs and pelvis were tender. He had a 3 x 4 cm tender and hard swelling over the upper part of his sternum. Another firm, non-tender swelling about 4 x 5 cm was seen in the right iliac region. Radiographs of the skull, spine and pelvis revealed multiple variable sized lytic lesions. A metastatic malignancy or disseminated tuberculosis was considered. His anti-tubercular therapy was intensified Fine needle aspiration from sternal lesion showed inflammatory cells. A bone marrow biopsy showed infiltration by tumor cells suggestive of metastatic adenocarcinoma. Patient's condition continued to deteriorate and he died within a fortnight of his hospitalization. CONCLUSION: Although masquerading as tuberculosis lytic lesions might be an evidence of malignant metastatic. Although, treatment is ineffective in this stage palliative efforts to improve quality of life should be made. PMID- 19134212 TI - Self-renewal and chemotherapy resistance of p75NTR positive cells in esophageal squamous cell carcinomas. AB - BACKGROUND: p75NTR has been used to isolate esophageal and corneal epithelial stem cells. In the present study, we investigated the expression of p75NTR in esophageal squamous cell carcinoma (ESCC) and explored the biological properties of p75NTR+ cells. METHODS: p75NTR expression in ESCC was assessed by immunohistochemistry. p75NTR+ and p75NTR- cells of 4 ESCC cell lines were separated by fluorescence-activated cell sorting. Differentially expressed genes between p75NTR+ and p75NTR- cells were determined by real-time quantitative reverse transcription-PCR. Sphere formation assay, DDP sensitivity assay, 64copper accumulation assay and tumorigenicity analysis were performed to determine the capacity of self-renewal, chemotherapy resistance and tumorigenicity of p75NTR+ cells. RESULTS: In ESCC specimens, p75NTR was found mainly confined to immature cells and absent in cells undergoing terminal differentiation. The percentage of p75NTR+ cells was 1.6%-3.7% in Eca109 and 3 newly established ESCC cell lines. The expression of Bmi-1, which is associated with self-renewal of stem cells, was significantly higher in p75NTR+ cells. p63, a marker identified in keratinocyte stem cells, was confined mainly to p75NTR+ cells. The expression of CTR1, which is associated with cisplatin (DDP) resistance, was significantly decreased in p75NTR+ cells. Expression levels of differentiation markers, such as involucrin, cytokeratin 13, beta1-integrin and beta4-integrin, were lower in p75NTR+ cells. In addition, p75NTR+ cells generated both p75NTR+ and p75NTR- cells, and formed nonadherent spherical clusters in serum-free medium supplemented with growth factors. Furthermore, p75NTR+ cells were found to be more resistant to DDP and exhibited lower 64copper accumulation than p75NTR- cells. CONCLUSION: Our results demonstrated that p75NTR+ cells possess some characteristics of CSCs, namely, self-renewal and chemotherapy resistance. Chemotherapy resistance of p75NTR+ cells may probably be attributable to decreased expression of CTR1. PMID- 19134215 TI - Horizontal Gene Transfers in prokaryotes show differential preferences for metabolic and translational genes. AB - BACKGROUND: Horizontal gene transfer (HGT) is an important process, which contributes in bacterial pathogenesis and drug resistance. A number of methods have been proposed for detection of horizontal gene transfer. One successful approach to the detection of HGT events is due to Novichkov et al. (J. Bacteriology 186, 6575-85), who rely on comparing phylogenetic distances within a gene family with genomic distances of the source organisms. Building on their approach, we introduce outlier detection in the correlation between those two sets of distances. This approach is designed to detect horizontal transfers of core set of genes present in many bacteria. The principle behind method allows detection of xenologous gene displacements as well as acquisition of novel genes. RESULTS: Simulations indicated that our method performs better than Novichkov et al's original approach. The approach very efficiently identified HGT between distantly related bacteria and also a limited number of gene transfers between closely related bacteria. In combination with sequence similarity and likelihood tests, it yields a measure robust enough to derive a set of 171 genes deemed likely to have been horizontally transferred. Further analysis of these 171 established horizontal transfer events gave interesting insights in the direction of transfer. CONCLUSION: The majority of transfers between archaea and bacteria have occurred in the direction from bacteria to archaea rather than the other way round. Genes transferred between the archaea and bacteria are mostly metabolic genes. On the other hand, genes transferred within the bacterial phyla are mainly involved in translation. PMID- 19134214 TI - Two distinct modes of nucleosome modulation associated with different degrees of dependence of nucleosome positioning on the underlying DNA sequence. AB - BACKGROUND: The nucleosome is the fundamental unit of eukaryotic genomes. Its positioning plays a central role in diverse cellular processes that rely on access to genomic DNA. Experimental evidence suggests that the genomic DNA sequence is one important determinant of nucleosome positioning. Yet it is less clear whether the role of the underlying DNA sequence in nucleosome positioning varies across different promoters. Whether different determinants of nucleosome positioning have characteristic influences on nucleosome modulation also remains to be elucidated. RESULTS: We identified two typical promoter classes in yeast associated with high or low dependence of nucleosome positioning on the underlying DNA sequence, respectively. Importantly, the two classes have low or high intrinsic sequence preferences for nucleosomes, respectively. The two classes are further distinguished by multiple promoter features, including nucleosome occupancy, nucleosome fuzziness, H2A.Z occupancy, changes in nucleosome positions before and after transcriptional perturbation, and gene activity. Both classes have significantly high turnover rates of histone H3, but employ distinct modes of nucleosome modulation: The first class is characterized by hyperacetylation, whereas the second class is highly regulated by ATP dependent chromatin remodelling. CONCLUSION: Our results, coupled with the known features of nucleosome modulation, suggest that the two distinct modes of nucleosome modulation selectively employed by different genes are linked with the intrinsic sequence preferences for nucleosomes. The difference in modes of nucleosome modulation can account for the difference in the contribution of DNA sequence to nucleosome positioning between both promoter classes. PMID- 19134216 TI - Rosiglitazone: can meta-analysis accurately estimate excess cardiovascular risk given the available data? Re-analysis of randomized trials using various methodologic approaches. AB - BACKGROUND: A recent and provocative meta-analysis, based on few outcome events, suggested that rosiglitazone increased cardiovascular mortality and myocardial infarction. However, results of meta-analyses of trials with sparse events, often performed when examining uncommon adverse effects due to common therapies, can vary substantially depending on methodologic decisions. The objective of this study was to assess the robustness of the rosiglitazone results by using alternative reasonable methodologic approaches and by analyzing additional related outcomes. FINDINGS: In duplicate and independently, we abstracted all myocardial and cerebrovascular ischemic events from all randomized controlled trials listed on the manufacturer's web site meeting inclusion criteria of the original meta-analysis (at least 24 weeks of rosiglitazone exposure in the intervention group and any control group without rosiglitazone). We performed meta-analyses of these data under different methodologic conditions. An unconfounded comparison that includes only trials (or arms of trials) in which medications apart from rosiglitazone are identical suggests higher risks than previously reported, making even the risk of cardiovascular death statistically significant. Alternatively, meta-analysis that includes all trials comparing a treatment arm receiving rosiglitazone to any control arm without rosiglitazone (as in the original meta-analysis) but also including trials with no events in both the rosiglitazone and control arms (not incorporated in the original meta analysis), shows adverse but non-statistically significant effects of rosiglitazone on myocardial infarction and cardiovascular mortality. Rosiglitazone appears to have inconsistent effects on a wider range of cardiovascular outcomes. It increases the risk of a broad range of myocardial ischemic events (not just myocardial infarction). However, its effect on cerebrovascular ischemic events suggests benefit, although far from statistically significant. CONCLUSION: We have shown that alternative reasonable methodological approaches to the rosiglitazone meta-analysis can yield increased or decreased risks that are either statistically significant or not significant at the p = 0.05 level for both myocardial infarction and cardiovascular death. Completion of ongoing trials may help to generate more accurate estimates of rosiglitazone's effect on cardiovascular outcomes. However, given that almost all point estimates suggest harm rather than benefit and the availability of alternative agents, the use of rosiglitazone may greatly decline prior to more definitive safety data being generated. PMID- 19134218 TI - Christ Siemens Touraine syndrome: a case report. AB - BACKGROUND: The ectodermal dysplasias are a large and complex group of diseases. CASE PRESENTATION: This article presents a case in a 37 years old female patient, referred to the dental clinic for impairment patients maintained by the University of Pernambuco. She presented typical characteristics of Christ Siemens Touraine syndrome such as alterations of the inferior members, a great number of diffuse pigmentations, poor oral hygiene, periodontal disease, oligodontia, enamel hypoplasia, including alteration in the form and size of the teeth. CONCLUSION: The optimal treatment for these patients should require the multidisciplinary collaborative efforts of health professionals. PMID- 19134219 TI - Concomitant dislocation of the tarsometatarsal and metatarsophalangeal joints of the second toe (floating second metatarsal): a case report. AB - When examining patients with injuries of the tarsometatarsal joint, the physician must pay attention to the foot as a whole. An extremely rare foot injury has been described in which axial and compressive forces cause simultaneous dislocation of the tarsometatarsal joint and the metatarsophalangeal joint of the same or adjacent ray. The following is a report of one of these rare injuries.We will also discuss probable mechanism and diagnosis of this rare traumatic injury. PMID- 19134217 TI - RNA interference-mediated c-MYC inhibition prevents cell growth and decreases sensitivity to radio- and chemotherapy in childhood medulloblastoma cells. AB - BACKGROUND: With current treatment strategies, nearly half of all medulloblastoma (MB) patients die from progressive tumors. Accordingly, the identification of novel therapeutic strategies remains a major goal. Deregulation of c-MYC is evident in numerous human cancers. In MB, over-expression of c-MYC has been shown to cause anaplasia and correlate with unfavorable prognosis. METHODS: To study the role of c-MYC in MB biology, we down-regulated c-MYC expression by using small interfering RNA (siRNA) and investigated changes in cellular proliferation, cell cycle analysis, apoptosis, telomere maintenance, and response to ionizing radiation (IR) and chemotherapeutics in a representative panel of human MB cell lines expressing different levels of c-MYC (DAOY wild-type, DAOY transfected with the empty vector, DAOY transfected with c-MYC, D341, and D425). RESULTS: siRNA mediated c-MYC down-regulation resulted in an inhibition of cellular proliferation and clonogenic growth, inhibition of G1-S phase cell cycle progression, and a decrease in human telomerase reverse transcriptase (hTERT) expression and telomerase activity. On the other hand, down-regulation of c-MYC reduced apoptosis and decreased the sensitivity of human MB cells to IR, cisplatin, and etoposide. This effect was more pronounced in DAOY cells expressing high levels of c-MYC when compared with DAOY wild-type or DAOY cells transfected with the empty vector. CONCLUSION: In human MB cells, in addition to its roles in growth and proliferation, c-MYC is also a potent inducer of apoptosis. Therefore, targeting c-MYC might be of therapeutic benefit when used sequentially with chemo- and radiotherapy rather than concomitantly. PMID- 19134220 TI - Canadian and English students' beliefs about waterpipe smoking: a qualitative study. AB - BACKGROUND: Waterpipe smoking is becoming popular among western students. The aim was to understand the appeal to students of this form of smoking when other forms of smoking are becoming less common. METHODS: Waterpipe smokers were identified by snowball sampling and interviewed following a semi-structured schedule in waterpipe cafes and in their homes. Constant comparative analysis was used to derive themes for the analysis. RESULTS: Waterpipe smokers saw smoking as an alternative to more expensive nights out in bars. The appeal was related to the communal activity and the novelty of the experience. Respondents had not thought deeply about the health risks and reasoned that if no warnings about waterpipe smoking were apparent (unlike cigarette smoking) then it was probably safe. These observations were reinforced by observations about the mildness of the smoke, the fruit flavours, and beliefs about the filtering of the water. Waterpipe smokers felt no pressure to stop smoking and therefore had not tried to do so, but felt it might be something they did not continue after university. Waterpipe smoking was not linked in students' minds to other forms of smoking except in one individual who was using waterpipe smoking to help quit cigarettes. CONCLUSION: In the absence of public health information, students have fallen back on superficial experiences to form views that waterpipe smoking is less harmful than other forms of smoking and it is currently much more acceptable in student society than other forms of smoking. PMID- 19134221 TI - Histological evaluation of the influence of magnetic field application in autogenous bone grafts in rats. AB - BACKGROUND: Bone grafts are widely used in oral and maxillofacial reconstruction. The influence of electromagnetic fields and magnets on the endogenous stimulation of target tissues has been investigated. This work aimed to assess the quality of bone healing in surgical cavities filled with autogenous bone grafts, under the influence of a permanent magnetic field produced by in vivo buried devices. METHODS: Metal devices consisting of commercially pure martensitic stainless steel washers and titanium screws were employed. Thirty male Wistar rats were divided into 3 experimental and 3 control groups. A surgical bone cavity was produced on the right femur, and a bone graft was collected and placed in each hole. Two metallic washers, magnetized in the experimental group but not in the control group, were attached on the borders of the cavity. RESULTS: The animals were sacrificed on postoperative days 15, 45 and 60. The histological analysis of control and experimental samples showed adequate integration of the bone grafts, with intense bone neoformation. On days 45 and 60, a continued influence of the magnetic field on the surgical cavity and on the bone graft was observed in samples from the experimental group. CONCLUSION: The results showed intense bone neoformation in the experimental group as compared to control animals. The intense extra-cortical bone neoformation observed suggests that the osteoconductor condition of the graft may be more susceptible to stimulation, when submitted to a magnetic field. PMID- 19134222 TI - Robust discovery of periodically expressed genes using the laplace periodogram. AB - BACKGROUND: Time-course gene expression analysis has become important in recent developments due to the increasingly available experimental data. The detection of genes that are periodically expressed is an important step which allows us to study the regulatory mechanisms associated with the cell cycle. RESULTS: In this work, we present the Laplace periodogram which employs the least absolute deviation criterion to provide a more robust detection of periodic gene expression in the presence of outliers. The Laplace periodogram is shown to perform comparably to existing methods for the Sacharomyces cerevisiae and Arabidopsis time-course datasets, and to outperform existing methods when outliers are present. CONCLUSION: Time-course gene expression data are often noisy due to the limitations of current technology, and may include outliers. These artifacts corrupt the available data and make the detection of periodicity difficult in many cases. The Laplace periodogram is shown to perform well for both data with and without the presence of outliers, and also for data that are non-uniformly sampled. PMID- 19134223 TI - Is new drug prescribing in primary care specialist induced? AB - BACKGROUND: Medical specialists are often seen as the first prescribers of new drugs. However, the extent to which specialists influence new drug prescribing in primary care is largely unknown. METHODS: This study estimates the influence of medical specialists on new drug prescribing in primary care shortly after market introduction. The influence of medical specialists on prescribing of five new drugs was measured in a cohort of 103 GPs, working in 59 practices, over the period 1999 until 2003. The influence of medical specialists on new drug prescribing in primary care was assessed using three outcome measures. Firstly, the proportion of patients receiving their first prescription for a new or reference drug from a specialist. Secondly, the proportion of GPs prescribing new drugs before any specialist prescribes to their patients. Thirdly, we compared the time until the GP's first own prescribing between GPs who waited for prescriptions from specialists and those who did not. RESULTS: The influence of specialists showed considerable differences among the new drugs studied. The proportion of patients receiving their first prescription from a specialist was greatest for the combination salmeterol/fluticasone (60.2%), and lowest for rofecoxib (23.0%). The proportion of GPs prescribing new drugs before waiting for prescriptions from medical specialists ranged from 21.1% in the case of esomeprazole to 32.9% for rofecoxib. Prescribing new drugs by specialists did not shorten the GP's own time to prescribing. CONCLUSION: This study shows that the influence of medical specialists is clearly visible for all new drugs and often greater than for the existing older drugs, but the rapid uptake of new drugs in primary care does not seem specialist induced in all cases. GPs are responsible for a substantial amount of all early prescriptions for new drugs and for a subpopulation specialist endorsement is not a requisite to initiate in new drug prescribing. This contradicts with the idea that the diffusion of newly marketed drugs always follows a two-step model, with medical specialists as the innovators and GPs as the followers. PMID- 19134224 TI - Conservation in the face of diversity: multistrain analysis of an intracellular bacterium. AB - BACKGROUND: With the recent completion of numerous sequenced bacterial genomes, notable advances have been made in understanding the level of conservation between various species. However, relatively little is known about the genomic diversity among strains. We determined the complete genome sequence of the Florida strain of Anaplasma marginale, and near complete (>96%) sequences for an additional three strains, for comparative analysis with the previously fully sequenced St. Maries strain genome. RESULTS: These comparisons revealed that A. marginale has a closed-core genome with few highly plastic regions, which include the msp2 and msp3 genes, as well as the aaap locus. Comparison of the Florida and St. Maries genome sequences found that SNPs comprise 0.8% of the longer Florida genome, with 33.5% of the total SNPs between all five strains present in at least two strains and 3.0% of SNPs present in all strains except Florida. Comparison of genomes from three strains of Mycobacterium tuberculosis, Bacillus anthracis, and Nessieria meningiditis, as well as four Chlamydophila pneumoniae strains found that 98.8%-100% of SNPs are unique to each strain, suggesting A. marginale, with 76.0%, has an intermediate level of strain-specific SNPs. Comparison of genomes from other organisms revealed variation in diversity that did not segregate with the environmental niche the bacterium occupies, ranging from 0.00% to 8.00% of the larger pairwise-compared genome. CONCLUSION: Analysis of multiple A. marginale strains suggests intracellular bacteria have more variable SNP retention rates than previously reported, and may have closed-core genomes in response to the host organism environment and/or reductive evolution. PMID- 19134225 TI - Crash characteristics and patterns of injury among hospitalized motorised two wheeled vehicle users in urban India. AB - BACKGROUND: Traffic crashes and consequent injuries represent a growing public health concern in India, particularly in light of increasing motorization. Motorised two-wheeled vehicles (MTV) constitute a large portion of the vehicle fleet in India. We report the crash characteristics and injury patterns among a cohort of MTV riders and pillions presenting to hospital post-crash. METHODS: Consecutive MTV riders and pillions, whether alive or dead, injured in a road traffic crash presenting to the emergency departments of two government hospitals and three branches of a private hospital in urban Hyderabad, India, were recruited to this study. RESULTS: 378 MTV users were enrolled to the study of whom 333 (88.1%) were male, 252 (66.7%) were riders and median age was 31.3 years. A total of 223 (59%) MTV users were injured in multi-vehicle crashes while one-third had a frontal impact. The majority (77%) were assessed as having a Glasgow coma score (GCS) of 13-15, 12% a GCS of 9-12 and 11% a GCS of 3-8. No difference was seen in the severity distribution of injuries based on GCS among riders and pillions. Open wounds and superficial injuries to the head (69.3%) and upper extremity (27%) and lower extremity (24%) were the most common injuries. 43 (11%) sustained an intracranial injury, including 12 (28%) with associated fracture of the bones of the head. There were few differences in types of injuries sustained by riders and pillions though riders had a significantly lower risk of crush injuries of the lower extremity than pillions (relative risk, RR 0.25, 95% CI 0.08-0.81) and female pillions were at a significantly lower risk of sustaining fractures of the lower extremity than male pillions (RR 0.30, 95% CI 0.09 - 0.94). Overall, 42 (11%) MTV users died, of which 42.8% died before reaching the hospital. Only 74 (19.6%) MTV users had worn a helmet correctly and failure to wear a helmet was associated with a five times greater risk of intracranial injury (RR 4.99, 95% CI 1.23-20.1). Of the 19 pre-hospital deaths, 16 (84%) had not worn a helmet. CONCLUSION: Head injuries accounted for the major proportion of injuries sustained in MTV users. Non-helmet use was associated with increased risk of serious head injuries. The data presented on the nature and severity of injuries sustained by MTV users can assist with planning to deal with these consequences as well as prevention of these injuries given the high use of MTV in India. PMID- 19134226 TI - Cell and molecular mechanisms of keratinocyte function stimulated by insulin during wound healing. AB - BACKGROUND: Regenerative wound repair is a goal of modern medicine. This is important not only for the local repair but also for its beneficial effect to systemic physiological processes. When wounds become chronic, individuals are susceptible to generalized inflammatory cascades that can affect many organs and even lead to death. Skin is the most commonly injured tissue, and its proper repair is important for reestablishment of its barrier function. RESULTS: We show here that insulin, when topically applied to skin excision wounds, accelerates re epithelialization and stimulates "maturation" of the healing tissue. These effects are dependent on the insulin receptor but independent of EGF/EGF-R; PI3K Akt-Rac1 signaling pathways are critically involved, and healing is alpha3 and LN332-dependent. CONCLUSION: Insulin has great potential for the treatments of chronic wounds in which re-epthelialization is impaired. Understanding of the pathways induced by insulin is important for the development of analog molecules that function strictly in healing. Because of its long history of safe use in humans for decades, this protein may prove to be a powerful therapy without major adverse effects. PMID- 19134227 TI - PreP+07: improvements of a user friendly tool to preprocess and analyse microarray data. AB - BACKGROUND: Nowadays, microarray gene expression analysis is a widely used technology that scientists handle but whose final interpretation usually requires the participation of a specialist. The need for this participation is due to the requirement of some background in statistics that most users lack or have a very vague notion of. Moreover, programming skills could also be essential to analyse these data. An interactive, easy to use application seems therefore necessary to help researchers to extract full information from data and analyse them in a simple, powerful and confident way. RESULTS: PreP+07 is a standalone Windows XP application that presents a friendly interface for spot filtration, inter- and intra-slide normalization, duplicate resolution, dye-swapping, error removal and statistical analyses. Additionally, it contains two unique implementation of the procedures - double scan and Supervised Lowess-, a complete set of graphical representations - MA plot, RG plot, QQ plot, PP plot, PN plot - and can deal with many data formats, such as tabulated text, GenePix GPR and ArrayPRO. PreP+07 performance has been compared with the equivalent functions in Bioconductor using a tomato chip with 13056 spots. The number of differentially expressed genes considering p-values coming from the PreP+07 and Bioconductor Limma packages were statistically identical when the data set was only normalized; however, a slight variability was appreciated when the data was both normalized and scaled. CONCLUSION: PreP+07 implementation provides a high degree of freedom in selecting and organizing a small set of widely used data processing protocols, and can handle many data formats. Its reliability has been proven so that a laboratory researcher can afford a statistical pre-processing of his/her microarray results and obtain a list of differentially expressed genes using PreP+07 without any programming skills. All of this gives support to scientists that have been using previous PreP releases since its first version in 2003. PMID- 19134228 TI - A foodborne outbreak of Cryptosporidium hominis infection. AB - Foodborne outbreaks of cryptosporidiosis are uncommon. In Denmark human cases are generally infrequently diagnosed. In 2005 an outbreak of diarrhoea affected company employees near Copenhagen. In all 99 employees were reported ill; 13 were positive for Cryptosporidium hominis infection. Two analytical epidemiological studies were performed; an initial case-control study followed by a cohort study using an electronic questionnaire. Disease was associated with eating from the canteen salad bar on one, possibly two, specific weekdays [relative risk 4.1, 95% confidence interval (CI) 2.1-8.3]. Three separate salad bar ingredients were found to be likely sources: peeled whole carrots served in a bowl of water, grated carrots, and red peppers (in multivariate analysis, whole carrots: OR 2.1, 95% CI 1.1-4.0; grated carrots: OR 2.1, 95% CI 1.2-3.9; peppers: OR 3.3, 95% CI 1.7-6.6). We speculate that a person excreting the parasite may have contaminated the salad buffet. PMID- 19134229 TI - Outbreaks of Shigella sonnei infections in Denmark and Australia linked to consumption of imported raw baby corn. AB - We investigated an outbreak of Shigella sonnei infections in Denmark and Australia associated with imported baby corn from one packing shed in Thailand. We reviewed nationwide surveillance and undertook case finding, food trace-back and microbiological investigation of human, food and environmental samples. A recall of baby corn and sugar snaps was based on descriptive epidemiological evidence. In Denmark, we undertook a retrospective cohort study in one workplace. In total, 215 cases were laboratory-confirmed in Denmark, and 12 in Australia. In a multivariable analysis, baby corn was the only independent risk factor. Antibiotic resistance and PFGE outbreak profiles in Denmark and Australia were indistinguishable, linking the outbreaks. Although we did not detect S. sonnei in baby corn, we isolated high levels of other enteric pathogens. We identified a packing shed in Thailand that supplied baby corn to Denmark and Australia, and uncovered unhygienic practices in the supply chain. This outbreak highlights the importance of international communication in linking outbreaks and pinpointing the source. PMID- 19134230 TI - Reported foodborne outbreaks due to noroviruses in Belgium: the link between food and patient investigations in an international context. AB - The Belgian data for foodborne norovirus (NoV) outbreaks became available for the first time with the introduction of an extraction and detection protocol for NoV in the National Reference Laboratory for foodborne outbreaks in September 2006. In 2007, 10 NoV foodborne outbreaks were reported affecting 392 persons in Belgium. NoV became the most detected agent in foodborne outbreaks followed by Salmonella (eight foodborne outbreaks). The major implicated foods were sandwiches (4/10), where food handlers reported a history of gastroenteritis in two outbreaks. A food handler was implicated in the limited number of Belgian NoV outbreaks which is in accord with internationally recorded data. Forty foodborne and waterborne outbreak events due to NoV, epidemiological and/or laboratory confirmed, from 2000 to 2007 revealed that in 42.5% of the cases the food handler was responsible for the outbreak, followed by water (27.5%), bivalve shellfish (17.5%) and raspberries (10.0%). PMID- 19134231 TI - The contribution of language skills to reading fluency: a comparison of two orthographies for Hebrew. AB - The purpose of the present study was to explore the contribution of phonological and general language skills to reading fluency of pointed and unpointed Hebrew scripts. Reading, language and memory tasks were performed by 48 fifth-grade monolingual native Hebrew speakers. Results showed that the most marked predictor for both pointed and unpointed reading texts was the morphological measure, whereas the phonological awareness measure contributed to neither of them. The semantic and syntactic measures contributed only to unpointed text reading fluency. The discussion highlights how readers in script, such as unpointed Hebrew, rely on general language skills in order to achieve fluent reading. PMID- 19134232 TI - Estimated mortality and hospital admission due to rotavirus infection in the WHO European region. AB - In 2006 two rotavirus vaccines were licensed for use in young children in Europe. This study aimed to estimate the mortality and hospital admissions due to rotavirus in children aged <5 years in the WHO European region using data from routine sources and published literature. We grouped 49/52 countries in the region by their World Bank Gross National Income (GNI) per capita. We obtained for children aged <5 years: populations, hospital discharges for diarrhoeal disease, estimated mortality rates and the percentage of deaths attributable to diarrhoeal disease, from WHO data sources or published literature, and combined them to estimate country-specific diarrhoeal disease mortality. Rotavirus attributable percentages of hospital admissions due to diarrhoeal disease were obtained through a literature search, and an income-group median applied to countries in each GNI category. In the countries we studied in the WHO European region, rotavirus infection causes an estimated 6550 deaths (range 5671-8989) and 146 287 (range 38 374-1 039 843) hospital admissions each year in children aged <5 years. Hospital admission rates were similar across income groups (medians 2.0, 2.8, 4.2 and 1.9/1000 per year in low-, lower-middle-, upper-middle- and high-income countries, respectively). Seven countries, mostly in the low- and lower-middle-income groups, accounted for 93% of estimated deaths. Disease burden varied dramatically by income level in the European region. Rotavirus vaccination in Azerbaijan, Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, Uzbekistan and Turkey could potentially prevent 80% of all regional rotavirus deaths. Data from low-income countries is still sparse, and improved disease burden studies are required to better inform regional vaccine policy. PMID- 19134233 TI - Bloodstream yeast infections: a 15-month survey. AB - A 15-month survey of 412 bloodstream yeast isolates from 54 Belgian hospitals was undertaken. Candida albicans was the most common species (47.3%) followed by C. glabrata (25.7%), C. parapsilosis (8.0%), C. tropicalis (6.8%) and Saccharomyces cerevisiae (5.1%). Common predisposing factors were antibacterial therapy (45%), hospitalization in intensive care units (34%), presence of in-dwelling catheters (32%), underlying cancer (23%) and major surgery (11%). Most patients had more than one predisposing factor. Fluconazole alone or in combination with another antifungal agent was the treatment of choice for 86.6% of the cases. Susceptibility testing revealed that 93.5% were susceptible to amphotericin B, 39.6% to itraconazole, 42.8% to fluconazole and 87% to voriconazole. Resistance to azoles was more common among C. glabrata isolates. PMID- 19134234 TI - Markov modelling of HIV infection evolution in the HAART era. AB - The aim was to investigate the impact of the main prognostic factors on HIV evolution. A multi-state Markov model was applied in a cohort of 2126 patients to estimate impact of these factors on patients' clinical and immunological evolutions. Clinical progression and immunological deterioration shared most of their prognostic factors: male gender, intravenous drug use, weight loss, low haemoglobin level (<110 g/l), CD8 cell count (<500/mm(3)) and HIV viral load (>5 log(10) copies/ml). Highly active retroviral therapy reduced the risks of clinical progression and immune deterioration whatever patients' CD4 cell count. Risk reductions were 41-60% for protease inhibitor-based and 27-68% for non nucleoside reverse transcriptase inhibitor-based regimens. Three-year transition probabilities showed that only patients with a CD4 cell count >or=350 CD4/mm(3) could in most cases maintain their immunity. This model provides 'real life' transition probabilities from one immunological stage to another, allowing decision analyses that could help determine the beneficial therapeutic strategies for HIV-infected patients. PMID- 19134235 TI - Impact of insecticide interventions on the abundance and resistance profile of Aedes aegypti. AB - Insecticide-based vector control is the primary strategy for curtailing dengue transmission. We used a mathematical model of the seasonal population dynamics of the dengue mosquito vector, Aedes aegypti, both to assess the effectiveness of insecticide interventions on reducing adult mosquito abundance and to predict evolutionary trajectories of insecticide resistance. We evaluated interventions that target larvae, adults, or both. We found that larval control and adult control using ultra-low-volume insecticide applications can reduce adult mosquito abundance with effectiveness that depends on the frequency of applications. We also found that year-long continuous larval control and adult control, using either insecticide treatment of surfaces and materials or lethal ovitraps, imposed the greatest selection for resistance. We demonstrated that combined targeting of larvae and adults at the start of the dengue season is optimal. This intervention contrasts with year-long continuous larval control policies adopted in settings in which dengue transmission occurs. PMID- 19134236 TI - Is Rotavirus contributing to an increase of diarrhoea in a region of Spain? AB - Diarrhoeal illnesses are the most frequent of notifiable diseases in Aragon. Physicians notify diarrhoea cases with presumed infectious origin on a weekly basis. Following an increase in 2005-2006, we aimed to identify the responsible organism(s) in order to inform control measures. We described seasonality of diarrhoea notifications for 1998-2004 and 2005-2006. We calculated correlations between diarrhoea notifications and enteric pathogens diagnosed in two Aragonese laboratories, and applied linear regression using coefficients of determination (r2). In 2005-2006 the winter peak of diarrhoea notifications increased from 2494 to 3357 weekly cases (34.6%) and the peak in Rotavirus diagnoses from 15 to 39 weekly cases. The correlation of diarrhoea notifications with Rotavirus was 0.05 in 1998-2004 and 0.42 in 2005-2006. The model for 1998-2004 included Salmonella enterica, Giardia lamblia and Clostridium difficile (r2=0.08) and for 2005-2006 Rotavirus and Astrovirus (r2=0.24). Our results suggest that Rotavirus contributed to the increase of diarrhoea notifications. We recommend determining the disease burden of Rotavirus in order to guide vaccination policies. PMID- 19134237 TI - Human bocavirus respiratory infections in children. AB - Human bocavirus (HBoV) was recently identified in respiratory samples from patients with acute respiratory infections and has been reported in different regions of the world. To the best of our knowledge, HBoV has never been reported in respiratory infections in Brazil. Nasopharyngeal aspirates were collected from patients aged <5 years hospitalized in 2005 with respiratory infections in Ribeirao Preto, southeast Brazil, and tested by polymerase chain reaction (PCR) for HBoV. HBoV-positive samples were further tested by PCR for human respiratory syncytial virus, human metapneumovirus, human coronaviruses 229E and OC43, human influenza viruses A and B, human parainfluenza viruses 1, 2 and 3, human rhinovirus and human adenovirus. HBoV was detected in 26/248 (10.5%) children of which 21 (81%) also tested positive for other respiratory viruses. Despite the high rates of co-infections, no significant differences were found between HBoV positive patients with and without co-infections with regard to symptoms. PMID- 19134238 TI - Farm-level risk factors for fluoroquinolone resistance in E. coli and thermophilic Campylobacter spp. on finisher pig farms. AB - Logistic regression, supported by other statistical analyses was used to explore the possible association of risk factors with the fluoroquinolone (FQ)-resistance status of 108 pig finisher farms in Great Britain. The farms were classified as 'affected' or 'not affected' by FQ-resistant E. coli or Campylobacter spp. on the basis of isolation of organisms from faecal samples on media containing 1 mg/l FQ. The use of FQ was the most important factor associated with finding resistant E. coli and/or Campylobacter, which were found on 79% (FQ-resistant E. coli) and 86% (FQ-resistant Campylobacter) of farms with a history of FQ use. However, resistant bacteria were also found on 19% (FQ-resistant E. coli) and 54% (FQ resistant Campylobacter) of farms with no history of FQ use. For FQ-resistant E. coli, biosecurity measures may be protective and there was strong seasonal variation, with more farms found affected when sampled in the summer. For FQ resistant Campylobacter, the buying-in of grower stock may increase risk and good on-farm hygiene may be protective. The findings suggest that resistant organisms, particularly Campylobacter, may spread between pig farms. PMID- 19134239 TI - Dietary patterns among a national sample of British children aged 1 1/2-4 1/2 years. AB - OBJECTIVE: Using a nationally representative sample, to identify groups among British children aged 1 1/2-4 1/2 years who report similar patterns of diet. DESIGN: Nationally representative dietary survey, using 4 d weighed dietary records, of girls and boys aged 1 1/2-4 1/2 years living in private households in Great Britain in 1992-1993. Cluster analysis was used to aggregate individuals into diet groups. SETTING: Great Britain. PARTICIPANTS: Eight hundred and forty eight boys and 827 girls. RESULTS: Three clusters were identified for girls and three for boys. Among boys the most prevalent cluster was 'Healthy Diet' (52.3%), the second was 'Convenience Diet' (38.3%) and the third was 'Traditional Diet' (9.3%). Among girls, the most prevalent dietary cluster was 'Healthy Diet' (58.7%), followed by a 'Convenience Diet' (36.6%) and 'Traditional Diet' (4.3%). There were important differences in nutrient profile, sociodemographic and behavioural characteristics between clusters. CONCLUSIONS: Cluster analysis identified three groups among both girls and boys which differed not only in terms of reported dietary intake, but also with respect to nutrient intake, social and behavioural characteristics. The groups identified could provide a useful basis for the development, monitoring and targeting of public health nutrition policy for pre-school children in the UK. Further research is needed on the consequences for chronic disease in the future for these children. PMID- 19134240 TI - Safety of supplementing infant formula with long-chain polyunsaturated fatty acids and Bifidobacterium lactis in term infants: a randomised controlled trial. AB - Probiotics and long-chain PUFA (LC-PUFA) may be beneficial supplements for infants who are not breast-fed. The aim of the present study is to evaluate the safety of an infant formula containing the LC-PUFA DHA and arachidonic acid (AA) and the probiotic Bifidobacterium lactis by comparing the growth rate of infants fed the supplemented and unsupplemented formulas. One hundred and forty-two healthy, term infants were enrolled in a single-centre, randomised, double-blind, controlled, parallel-group trial, and allocated to receive either standard or probiotic and LC-PUFA-containing experimental formulas. The infants were fed with their assigned formulas for 7 months. The primary outcome (weight gain) and the secondary outcomes (length, head circumference and formula tolerance) were measured throughout the study. LC-PUFA status was assessed at 4 months of age and immune response to childhood vaccines was measured at 7 months of age. There was no significant difference in growth between the two groups. The 90 % CI for the difference in mean weight gain was - 0.08, 3.1 g in the intention-to-treat population and 0.1-3.8 g in the per protocol population, which lay within the predefined boundaries of equivalence, - 3.9-3.9. There were no significant differences in mean length and head circumference. DHA and AA concentrations were higher in infants in the experimental formula group compared with the control formula group. No influence of the supplements on the response to vaccines was observed. Growth characteristics of term infants fed the starter formula containing a probiotic and LC-PUFA were similar to standard formula-fed infants. PMID- 19134241 TI - Hypobaric pressure exposure effects on cochlear frequency selectivity in fluctuating, low-frequency hearing loss. AB - AIM: To study the effects of hypobaric pressure chamber exposure on the cochlear frequency selectivity of subjects with monaural, fluctuating, low-frequency hearing loss, such as occurs in Meniere's disease. METHODS: We used a hypobaric pressure chamber to create relative underpressure in the ear canal, in order to impose positive pressure gradients on the inner ear. Psychophysical tuning curves, transiently evoked otoacoustic emissions and speech recognition scores in noise were measured in 10 subjects with fluctuating, low-frequency hearing loss, before and after pressure exposure. RESULTS: After the exposure, subjects' overall pure tone averages showed no improvement, but individual results showed improved speech recognition scores in noise (six subjects), increased transiently evoked otoacoustic emission strength (three subjects) and increased psychophysical tuning curve steepness (two subjects). Deteriorations were also seen, mainly in psychophysical tuning curves. No association could be established between the different tests, and the measured parameters could not predict subjective improvement. CONCLUSION: The results suggest variable effects of hypobaric pressure exposure on inner-ear physiology. PMID- 19134242 TI - Effect of phosphodiesterase-5 inhibitor on hearing. AB - OBJECTIVE: Following a report of sudden hearing loss in a patient taking phosphodiesterase type 5 inhibitor, and a Food and Drug Administration announcement concerning this class of drugs, a study was planned to investigate if ototoxicity occurs in patients using phosphodiesterase 5 inhibitor for erectile dysfunction. METHODS: Eighteen patients with erectile dysfunction who had been using phosphodiesterase 5 inhibitor were included in the study. Audiometric tests were performed on all patients, between the frequencies 250 and 16,000 Hz, before and 1, 5 and 72 hours after drug ingestion. RESULTS: Four patients showed a unilateral threshold decrease compatible with ototoxicity criteria; this change was reversible. A statistically significant difference in pre- versus post-drug hearing thresholds was observed in the right ear at 10,000 Hz (p = 0.008). There were no statistically significant hearing threshold differences at any other frequencies (p > 0.05). CONCLUSION: Although temporary ototoxicity was noted in four patients, we could not find any permanent, deleterious effect of phosphodiesterase type 5 inhibitor on hearing thresholds. PMID- 19134243 TI - Sutter bipolar radiofrequency volumetric tissue reduction of palate for snoring and mild obstructive sleep apnoea: is one treatment adequate? AB - OBJECTIVE: To evaluate the efficacy of bipolar radiofrequency volumetric tissue reduction, using Sutter technology, in the treatment of snoring and mild obstructive sleep apnoea. DESIGN: Prospective, non-randomised trial. METHODS: Fifty-two patients with simple snoring and mild obstructive sleep apnoea underwent bipolar radiofrequency palate reduction under local anaesthesia. RESULTS: All patients were Friedman stage II and III, with tonsil sizes graded as zero, one or two, a mean body mass index of 22.6, and a mean age of 36.2 years. Seventy-seven per cent of patients reported improvement in their snoring; patients' mean snoring level (assessed by visual analogue scale) improved from 8.9 to 3.4 (p < 0.05). Eighty-three per cent of patients reported an improvement in their Epworth sleepiness scale score, from a mean of 14.6 to 9.5. Seven out of the 17 patients (41.2 per cent) met the surgical success criteria (i.e. at least a 50 per cent reduction of the pre-procedure apnoea-hypopnoea index, and a post procedure apnoea-hypopnoea index of below 15). Patients' mean apnoea-hypopnoea index improved from 13.6 to 9.8, and their mean lowest oxygen saturation improved from 88.3 to 92.5 per cent. Eighty-two per cent of patients reported an improvement in their quality of life as a result of the procedure. CONCLUSION: One session of Sutter bipolar radiofrequency tissue volume reduction represents a viable method of treating snoring and mild obstructive sleep apnoea, with good results. PMID- 19134244 TI - Sensitivity and specificity of computed tomography for detection of extranodal spread from metastatic head and neck squamous cell carcinoma. AB - AIM: To estimate the sensitivity and specificity of computed tomography used for the detection of extranodal spread of metastatic head and neck squamous cell carcinoma, by experienced head and neck radiologists. MATERIALS AND METHODS: Participants had undergone a neck dissection for head and neck squamous cell carcinoma, together with computed tomography scanning prior to surgery (accessible for reporting). Computed tomography images were independently examined by two experienced head and neck radiologists. Nodal involvement by squamous cell carcinoma and the presence or absence of extranodal spread were recorded. Results were compared to the histological specimen. The sensitivity, specificity and positive predictive value of using computed tomography for the detection of nodal involvement and presence or absence of extranodal spread were estimated, and 95 per cent confidence intervals were calculated. RESULTS AND ANALYSIS: The study analysed 149 neck dissections. When using computed tomography to detect the extranodal spread of head and neck squamous cell carcinoma, radiologists A and B had sensitivities of 66 and 80 per cent, specificities of 91 and 90 per cent, and positive predictive values of 85 and 87 per cent, respectively. DISCUSSION: The sensitivity and specificity of radiological detection of extranodal spread from head and neck squamous cell carcinoma is not well reported in the literature. Accuracy of reporting improves in the hands of experienced head and neck radiologists. This finding has clinical implications for surgical planning and adjuvant therapy requirements. PMID- 19134245 TI - Myringoplasty: impact of perforation size on closure and audiological improvement. AB - OBJECTIVES: To investigate the impact of perforation size and other variables on the success of myringoplasty, and also to determine audiological gain following successful closure of tympanic membrane perforations of various sizes. STUDY DESIGN: Retrospective analysis of 130 case notes. MAIN OUTCOME MEASURES: (1) Successful closure of tympanic membrane following myringoplasty, in relation to recorded variables (i.e. perforation size, grade of surgeon, surgical technique, graft material, previous myringoplasty and smoking history). (2) Mean, four frequency, air conduction audiometric gain following successful myringoplasty for various, pre-operatively categorised tympanic membrane perforation sizes. RESULTS AND CONCLUSION: The collective myringoplasty success rate was 80.8 per cent (105/130); for successful patients, the mean air conduction audiometric gain was 6.8 dB (t = 5.29, p < 0.0001). Neither perforation size nor any other assessed variable was a statistically significant determinant factor for successful myringoplasty. Air conduction audiometric gains following successful myringoplasty were directly correlated with pre-operative perforation size (-4.0 dB for 0-20 per cent perforations, -5.0 dB for 21-40 per cent, -9.1 dB for 41-60 per cent, -10.8 dB for 61-80 per cent and -13.3 dB for 81-100 per cent). PMID- 19134246 TI - Safety of intravenous use of ketorolac in infants following cardiothoracic surgery. AB - OBJECTIVES: To evaluate the impact of intravenous ketorolac on renal function and haematologic values in patients less than six months old following cardiothoracic surgery. BACKGROUND: Ketorolac is a potent nonsteroidal anti-inflammatory drug indicated for short term management of moderate to severe post-operative pain. Little data is available related to its safety in infants less than six months of age. METHODS: This was a retrospective, case-control chart review of 19 patients aged less than six months of age with biventricular circulations who received intravenous ketorolac following cardiothoracic surgery. They were compared with 19 age-matched control patients. Those with functionally univentricular anatomy were excluded due to their higher risk for renal impairment following surgery. Student's t-test was used to compare the incidence of renal impairment and haematologic complications between the groups, as well as the number of analgesic doses administered. Charts were reviewed for number of blood transfusions. RESULTS: Patients receiving intravenous ketorolac had no statistically significant changes in pre-operative versus post-treatment renal function or haematologic effects compared to the control group. No statistically significant differences were detected for number of post-operative blood transfusions or additional analgesic administration between groups. CONCLUSIONS: Intravenous ketorolac appears to be safe when used in infants less than six months of age with biventricular circulations following cardiothoracic surgery. Ketorolac as used in these patients does not decrease the use of standard analgesic therapy. PMID- 19134247 TI - [Clinical diagnosis of anomalous origin of the left coronary artery from the pulmonary artery in 22 cases]. AB - OBJECTIVE: To discuss the diagnostic methods and features of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA). METHODS: A total of 22 cases of ALCAPA hospitalized between 2000 and 2007 were recruited and divided into infant group (age < or = 1 year, n = 10) and older children group (age > 1 year, n = 12). The patients' history, electrocardiography (ECG) and echocardiography were reviewed and analyzed retrospectively. ECGs were analyzed as follows: (1) Q wave and T wave inversion in lead I, (2) Abnormal Q wave and T wave inversion in lead aVL, (3) Q wave in lead V(5-6), (4) T wave inversion and ST changes in lead V(4-6), (5) LV hypertrophy. Echocardiograms were analyzed as follows: (1) Continuity of the left coronary artery (LCA) and pulmonary artery (PA), (2) Retrograde shunt into PA, (3) Increased papillary muscle echodensity, (4) Right coronary artery (RCA) dilation, (5) Collateral signals within the ventricular septum. RESULTS: The presence of cardiomegaly in X-ray film (18/22), aVL QT pattern in ECG (17/22), retrograde color Doppler flow into pulmonary artery (20/22), anterior lateral papillary echogenic (17/22) and collateral vessel signals (16/22) in echocardiography were high in both groups (P > 0.05). The presence of clinical symptoms and abnormal Q wave in leads Iand V(5-6) in ECG were significantly higher in the infant group than in the older children group (P < 0.05). But the presence of right coronary artery dilation was significantly lower in the infant group than in the older children group (P < 0.05). CONCLUSION: Different diagnostic features were found in infant and older children patients. With combination of patient history, electrocardiogram and echocardiogram, accurate diagnosis could be obtained in most pediatric patients with ALCAPA. PMID- 19134248 TI - [Clinical and electrophysiologic studies on epileptic negative myoclonus in atypical benign partial epilepsy of childhood]. AB - OBJECTIVE: To investigate the clinical, neurophysiologic characteristics and therapeutic considerations of epileptic negative myoclonus (ENM) in atypical benign partial epilepsy of childhood (ABPE). METHODS: Video-EEG monitoring with outstretched arm tests were carried out in 17 patients, and 9 of them were examined with simultaneous electromyography (EMG). The ENM manifestations, electrophysiologic features and responses to antiepileptic drugs (AED) were analyzed. RESULTS: Seventeen patients were diagnosed as having benign childhood epilepsy with centrotemporal spikes (BECT) during the early course of the disease and were treated with AED. During the course of the disease, hand trembling, objects dropping, head nodding and instability during standing might be clues for ENM occurrence. ENM had been confirmed in our patients by outstretched arm tests during video-EEG recording. The ictal EEG showed that high-amplitude spikes followed by a slow wave over the contralateral motor areas. This was further confirmed by time-locked silent EMG in 9 patients. During ENM occurrence or recurrence, the habitual seizures and interictal discharges were exaggerated. Atypical absence seizures also occurred in 6 patients. The alteration of therapeutic options of AED relating to ENM appearance in some patients included the add-on therapy with carbamazepine (CBZ), oxcarbazepine, phenobarbital, or withdrawal of valproate (VPA). ENM was controlled in most cases by using VPA, clonazepam (CZP) and corticosteroid with different combination. CONCLUSION: ENM could occur during the course of ABPE. Outstretching arm tests during video-EEG monitoring in combination with EMG was essential to confirm ENM. The ENM occurrence was always associated with the frequency increasing of habitual seizures and the aggravation of interictal discharges. Some AED such as CBZ might induce ENM. VPA, benzodiazepines and corticosteroid with different combination were relatively effective in treatment of ENM. PMID- 19134249 TI - [Relationship between clinical pathophysiology and pulmonary pathology in patients with congenital heart defects and decreased pulmonary artery blood flow]. AB - OBJECTIVE: To investigate the relationship between pulmonary pathological features and clinical physiology of congenital heart defects (CHD) with decreased pulmonary artery blood flow. METHODS: Between July 2001 and May 2006, 18 patients with CHD with decreased pulmonary artery blood flow undergoing palliative or definitive repair and having lung biopsy intraoperatively were enrolled in this study. The patients' age was 0.4 - 8.0 years, and body weight was 6.0 - 20.0 kg. The method of semi-quantitative morphometric technique and an image analyzer were applied to measure the following indices of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%) and numbers of microvessels per square centimeter (VPSC). The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two dimensional echocardiography. The percutaneous oxygen saturation (SpO(2)), hemoglobin concentration (HB) and hematocrit value (HCT) were examined and recorded preoperatively. RESULTS: There was a significant negative correlation between SpO(2) and HCT or Hb (R(2) = 0.4914, P = 0.001 and R(2) = 0.5505, P < 0.001), the variation trend of these three variables was linked. There was a negative correlation between SpO(2) and the body weight (R(2) = 0.2208, P = 0.049), which is in accordance with clinical features of aggravated process of cyanosis and hypoxia. The morphological observation of lung biopsy specimens indicated that most of peripheral pulmonary arteries were distended, irregular and their walls were uneven, and "lake" type of pulmonary AV malformations were observed. There was a positive correlation between VPSC and the body weight or BSA (R(2) = 0.5472, P < 0.001 and R(2) = 0.5233, P = 0.001). There was a significant correlation between VPSC and LPA or RPA (R(2) = 0.4312, P = 0.003 and R(2) = 0.2463, P = 0.036). It was shown that the diameter of central pulmonary arteries could be a reflection of peripheral pulmonary artery growth. The diameter of LPA also correlated with the diameter of RPA (R(2) = 0.286, P = 0.022). CONCLUSIONS: For patients with congenital heart defects with decreased pulmonary blood flow, the pulmonary pathological changes are the bases of their clinical physiologic features. It is suggested that they should be treated in their earlier stage of life. PMID- 19134250 TI - [Balloon dilation with gastroscope for esophageal stricture in children]. AB - OBJECTIVE: To evaluate the efficacy and safety of balloon dilation (BD) with gastroscope in treatment of esophageal stricture in children. METHODS: BD was performed in 12 children aged 5 - 59 months, average age 26 months, course of disease was 2 - 26 months, with esophageal stricture, 7 cases with anastomotic strictures secondary to surgical repair of esophageal atresia, 3 with congenital esophageal stenosis, 2 with corrosive esophageal strictures. All procedures were performed under tracheal intubation and intravenous anesthesia using the 3rd grade controlled radial expansion (CRE) esophagus-balloon with gastroscope. Firstly the balloon was inserted into the esophagus through mouth, then put in the gastroscope. Under the direct guidance of gastroscope the balloon was positioned across the stricture, then the balloon was filled with saline to get needed pressure and maintained for 3 minutes. The procedure was repeated 3 times at an interval of 3 minutes. The abdominal pain, melena and vomiting were observed, as well as the diet taken thereafter, the size of the stricture and the nutrition status were observed for 3 to 12 months after the dilation. RESULTS: Twenty-two dilations were performed in 12 cases, 19 succeeded, 3 cases developed complication during the dilation, the total success rate was 86%. The procedure failed in 3 cases and succeeded in 9 cases, the effective rate was 75%. Follow-up and repeated gastroscopy were performed within 3 to 12 months after the dilation, the diameter of the stricture was 9-13 mm, compared with 2-8 mm before the dilation. Eight of the children could take solid food and nutritional status was improved. CONCLUSIONS: BD with the 3rd grade CRE esophagus-balloon under gastroscopy is a simple and effective method to treat esophagus stricture in children, especially for anastomotic strictures secondary to surgical repair of esophageal atresia. PMID- 19134251 TI - [Molecular epidemiological study on norovirus among children with acute diarrhea in Guangzhou]. AB - OBJECTIVE: To study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments. METHODS: Fecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced. RESULTS: Totally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March). CONCLUSION: NV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain. PMID- 19134252 TI - [Effect of sulfur dioxide on vascular collagen remodeling in spontaneously hypertensive rats]. AB - OBJECTIVE: To study the modulatory effect of sulfur dioxide (SO(2)) on the accumulation of collagen type I and type III in the wall of aorta during spontaneously hypertensive rat (SHR) vascular remodeling. METHODS: Eight male Wistar Kyoto rats at the age of 4 weeks with normal blood pressure were used as a control group. And sixteen male SHRs at the age of 4 weeks were randomly divided into an SHR control group and SHR + Na(2)SO(3)/NaHSO(3) (SO(2) donor) group. Na(2)SO(3)/NaHSO(3) solution was injected intraperitoneally everyday to the rats in the SHR + Na(2)SO(3)/NaHSO(3) group. After 5 weeks, the systemic blood pressure was measured. The weight ratio of left ventricle to the whole heart was also measured. The rat aorta was dyed with Hart's method. The morphometric parameters including outer radius, lumen radius and the wall thickness were calculated by Leica workstation. The plasma level of SO(2) was determined by HPLC method. The expressions of collagen type I and type III in aorta were detected by immunohistochemistry. RESULTS: (1) Compared with the WKY rat control group, the systolic blood pressure increased by 53%, the weight ratio of left ventricle to the whole heart increased by 6% but the plasma level of SO(2) decreased by 44% for rats in the SHR group (P < 0.01 or P < 0.05). Compared with the SHR control group, the systolic blood pressure decreased by 26%, but the plasma level of SO(2) increased by 28% (all P < 0.01) for rats in the SHR + Na(2)SO(3)/NaHSO(3) group. (2) Compared with the WKY rats, the ratio of media to lumen radius increased by 28% for SHR. Compared with the SHR group, the ratio of media to lumen radius decreased by 10% (P < 0.01) in rats of the SHR + Na(2)SO(3)/NaHSO(3) group. (3) Compared with rats in the WKY control group, collagen type I expression increased by 10% for rats in the SHR group (P < 0.01). Compared with the SHR group, however, the expression decreased by 58% for rats in the SHR + Na(2)SO(3)/NaHSO(3) group (P < 0.01). (4) Compared with rats in the WKY control group, the expression increased by 13% for rats in the hypoxia SHR group (P < 0.01); however, compared with rats in the SHR group, the expression decreased by 8% in the rats of the SHR +Na(2)SO(3)/NaHSO(3) group (P < 0.01). CONCLUSIONS: In the process of SHR vascular collagen remodeling in the rats, SO(2) could inhibit the abnormal accumulation of collagen type I and type III in the wall of aorta. This effect may be one of the mechanisms by which SO(2) ameliorates SHR vascular remodeling. PMID- 19134253 TI - [Natural history of moderate aplastic anemia in children and effects of immunosuppressive therapy]. AB - OBJECTIVE: In contrast to severe aplastic anemia (SAA), the appropriate management of patients with moderate aplastic anemia (MAA) is unclear. Recently, it was reported that when childhood MAA was treated with supportive care alone, 2/3 of patients progressed to SAA, and therefore patients with MAA should be treated with immunosuppressive (IS) therapy in time. The present study aimed to review the natural history, the rate of progression to SAA and outcome of children with MAA seen at our institution over the past 12 years and to explore the relationship between the effectiveness of IS therapy and the immune mediated pathological mechanism. METHODS: Seventy-one MAA patients were included in this study. At the first stage, thirty-six children with MAA were given IS therapy (IS group, antithymocyte globulin, ATG or cyclosporin-A, CSA). The therapeutic effects were evaluated and compared with those of 35 children with MAA who received the treatment of supportive care alone (androgens, control group). At the second stage, the patients with MAA progressed to SAA were given combined immunosuppressive (CIS) therapy (CIS group, a combination of ATG, CSA and high dose immunoglobulin). Peripheral blood lymphocyte subsets levels were measured with a flow cytometer. RESULTS: At the first stage, in the IS group, the percentage of overall and complete responders was 83.3% and 69.4%, respectively, which was significantly higher than that of the control group (34.3% and 17.1%). Twenty-three patients with MAA progressed to SAA. In the control group, 18 patients with MAA progressed to SAA. In the IS group, five patients with MAA progressed to SAA. The 17 patients with MAA who progressed to SAA were given combined immunosuppressive therapy. The percentage of overall and complete responders was 70.6% and 41.2%, respectively. The level of CD4(+), NK cell ratio decreased but the level of CD8(+) cell increased in MAA children before the treatment. The level of NK and CD4(+) cell was significantly higher in the IS group with the treatment than in the control group. CONCLUSION: When childhood MAA is treated with supportive care alone, more than 50% of patients may progress to SAA. Immune mediated pathological mechanism of MAA might be the base of IS therapy. IS therapy is effective and safe for childhood MAA.CIS therapy given to patients with MAA that was progressed to SAA may also be effective. PMID- 19134254 TI - [Clinical and pathological study of 47 cases with Alport syndrome]. AB - OBJECTIVE: To analyze the clinical and pathological features of children with Alport syndrome (AS). METHODS: A series of 47 patients with AS from unrelated families hospitalized from Jan. 1990 to Jan. 2007 were involved in this study. The clinical and histopathological data were collected and analyzed. RESULTS: Of the 47 cases, 32 were male and 15 female, M/F: 2.1:1. The patient's age ranged from 15 months to 13 years, mean 9 years. Thirty-nine of the 47 cases had positive family history, X-linked dominant inheritance AS was diagnosed in 37 cases, autosomal recessive inheritance AS in 2 cases. Gross hematuria or microscopic hematuria were found in 59.3% of the cases as the first manifestations, while 29.8% showed edema or proteinuria. The major clinical manifestations were isolated hematuria (23.4%), hematuria and proteinuria (36.2%), nephrotic syndrome (29.8%), and renal failure (10.6%). Hematuria and proteinuria existed in all the cases, while only 7 to 13 years children had nephrotic syndrome and renal failure. Of the 47 patients, 33 (70.2%) showed mesangial proliferative glomerulonephritis (MsPGN) under the light microscope, 13 (27.6%) focal segmental glomerulosclerosis (FSGS), 1 (2.1%) membrane proliferative glomerulonephritis (MPGN). For immunofluorescence, there was IgM (40.4%) as the dominant deposition in 19 patients, IgA in 9 (19.1%), IgG in 9 (19.1%), and 10 (21.4%) were negative. Thirty-nine cases showed typical glomerular basement membrane (GBM) pathological changes under electron microscope, while thin basement membrane in 8 cases; 46 showed abnormal skin and/or renal alpha-chain distribution. CONCLUSION: For Alport syndrome, number of male patients was higher than that of female patients. There was a significant difference among different age groups. Hematuria might be present throughout the course, while urine protein increases gradually. MsPGN was the dominant pathological change. The GBM pathological changes in younger children is not typical, so the immunofluorescence test of alpha-chain in collagen IV should be used as an important diagnostic method. PMID- 19134255 TI - [Molecular analysis of survival motor neuron gene in 338 suspicious children patients with spinal muscular atrophy]. AB - OBJECTIVE: Spinal muscular atrophy (SMA) is an autosomal recessive disorder that results in symmetrical muscle weakness and wasting due to degeneration of the anterior horns of the spinal cord. The clinical picture of SMA is variable and childhood SMA has been classified into 3 types on the basis of the age of onset and clinical course. The survival motor neuron (SMN) gene was mapped to chromosome 5q13. The SMN1 gene has been recognized to be responsible for SMA because of homozygous deletions or intragenic mutations in SMN1 results in childhood onset of SMA. The main objective of this study was to determine the deletion frequency of SMN1 gene and to apply gene analysis in children patients with SMA. METHODS: The SMA patients were diagnosed and clinically typed according to the international diagnostic criteria, following up cases, and gene analysis. The PCR enzyme assay was used to detect the homozygous deletion of SMN1 gene in SMA patients. A dosage assay that combined multiplexed allele-specific PCR and DHPLC was used to determine the copy numbers of the SMN1 and SMN2 and detect SMN1 heterozygous deletion. RESULTS: (1) A total of 267 patients with SMA were diagnosed from 338 suspicious cases and 143, 82, and 42 cases were typed as types I, II, and III, with the percentages of 53.6% (143/267), 30.7% (82/267) and 15.7% (42/267), respectively. (2) Results of the present study showed that 68.5% (183/267) of SMA patients had homozygous deletions of exons 7 and 8 of SMN1 gene and 12.7% (34/267) had homozygous deletions of only exon 7 of SMN1 gene. The SMN1 heterozygous deletion was confirmed in 12.4% (33/267) of SMA patients. Non deletion SMA patients accounted for 6.4%(17/267). The homozygous deletions of only exon 8 of SMN1 gene could not be detected. (3) The rates of homozygous or heterozygous deletion in types I and II were very similar. The rate of homozygous deletion was lower in type III than that in type I or II and rate of heterozygous deletion of type III was higher than that in types I or II. CONCLUSION: (1) The frequency and pattern of deletions in the Chinese children patients with SMA are significantly different from that observed in Caucasians populations. Further gene characterization and subtle mutations within the SMN1 gene need to be studied in order to define the molecular basis of SMA in the Chinese population. (2) The gene diagnosis is a special and non invasive method as compared with other methods. A total of 80% patients can be diagnosed through the analysis of the homozygous deletion of SMN1 gene. (3) The clinical diagnosis and gene detection need to be studied in future for the SMA patients with type III. PMID- 19134256 TI - [Mechanism of conditioned immune response in curing bronchial asthma in mice]. AB - OBJECTIVE: To understand the mechanism of effect of conditioned immune response in curing bronchial asthma. METHODS: An experimental asthma modal was produced on healthy BALB/C mice (female, 4 - 6 weeks old) by sensitization and stimulation with ovalbumin (OV A). Totally 105 mice were divided into 7 groups randomly with 15 in each and treated differently: in group CIR(1), noise was used as conditioned stimulus (CS) and budesonide and salbutamol as unconditioned stimulus (UCS) respectively, a conditioned immune response model of mice with asthma was established by the combination of CS and UCS 7 times (7 days), then the mice were given CS only, and the combination were given once a week for 20 weeks. In group CIR(2) saccharin (SAC) was taken as CS, and the other treatments were the same as the group CIR(1). In the group of conventional therapy, the mice were given inhalation of nebulized budesonide and salbutamol only for 20 weeks. In the group of lower dose conventional therapy, the mice were given nebulized inhalation of budesonide and salbutamol for the first 7 days, then once a week for 20 weeks. In the noise group the mice were given noise only everyday for 20 weeks. In SAC group the mice were treated with SAC only everyday for 20 weeks. In the blank control group the mice were treated with placebo for 20 weeks. The mice in all the groups were stimulated with OVA once a day. The mice in the healthy control group were given PBS inhalation for 20 weeks. After 20 weeks therapy, the bronchoalveolar lavage fluid (BALF) was taken for eosinophils (EOS) counting. The spleens were taken to obtain CD4(+)T lymphocytes and the expression of neuronal acetylcholine receptor alpha 7 (nAChRalpha7), IL-4, IFN-gamma and IL-17 were detected by flow cytometry. RESULTS: (1) The percent of EOS of groups CIR(1), CIR(2), conventional therapy and healthy control was much lower than that of blank control (P < 0.01), and there was no significant difference among groups CIR(1), CIR(2) and conventional therapy (P > 0.05). (2) The expression of nAChRalpha7, IL-4 and IL-17 of groups CIR(1), CIR(2), conventional therapy and healthy control was much lower than that in blank control group, IFN-gamma was much higher (P < 0.01), and no significant difference was found among groups CIR(1), CIR(2) and conventional therapy (P > 0.05). There was a positive correlation between nAChRalpha7 and IL-4 (r = 0.76, P < 0.01), nAChRalpha7 and IL 17 (r = 0.46, P < 0.01). There was a negative correlation between nAChRalpha7 and IFN-gamma (r = 0.69, P < 0.01). (3) In the groups treated with lower dose of conventional therapy, noise, SAC and blank control, the epithelial tissue of airway were much thicker, the lumens were much narrower, and inflammatory cells and collagen fibers were much more than in the healthy control group, and after therapy, the inflammation in groups CIR(1), CIR(2) and conventional therapy was significantly improved. CONCLUSION: The conditioned immune response models established by both noise and SAC as CS and budesonide and salbutamol as UCS can downregulate nAChRalpha7 on CD4(+)T lymphocytes, regulate the function of CD4(+)T lymphocytes, and achieve the same therapeutic efficacy in treatment of asthma. PMID- 19134257 TI - [Effect of cell proliferation and OPN expression in rat myocardial fibroblasts infected with coxsackievirus B3]. PMID- 19134258 TI - [Complications and geometric changes in children after percutaneous closure of atrial septal defect]. PMID- 19134260 TI - [A case with Treacher-Collins syndrome]. PMID- 19134259 TI - [Coccidioidomycosis in an infant]. PMID- 19134261 TI - [Abernethy malformation in a case]. PMID- 19134262 TI - [Several problems in diagnosis and treatment of chronic cough in children: intensive reading of the guideline for diagnosis and treatment of chronic cough in pediatrics (III)]. PMID- 19134263 TI - [An updated classification of primary immunodeficiency diseases]. PMID- 19134264 TI - [Atypical hemolytic uremic syndrome and the CFH gene]. PMID- 19134265 TI - [Mechanism and clinical research of spontaneous closure of ventricular septal defect]. PMID- 19134266 TI - [Determination of coronary hemodynamic parameters by Doppler ultrasound in children with Kawasaki disease]. PMID- 19134268 TI - [The quality of genetic association studies need to be improved]. PMID- 19134269 TI - [The genotype-phenotype correlation of MYH7 gene G15391A mutation and MYBPC3 gene G12101A mutation in familial hypertrophic cardiomyopathy]. AB - OBJECTIVE: To reveal genotype-phenotype correlation of disease-causing gene mutations in Chinese hypertrophic cardiomyopathy (HCM) pedigree. METHODS: Peripheral venous blood samples were collected from two Chinese HCM families and 120 healthy subjects were recruited as normal control. The full encoding exons and flanking sequences of the cardiac troponin T gene (TNNT2), beta-myosin heavy chain gene (MYH7) and myosin binding protein C gene (MYBPC3) were amplified with the polymerase chain reaction method, DNA sequencing was used to detect the mutation. RESULTS: In ZZJ family, mutation G12101A was identified in exon 21 of MYBPC3 gene in 4 family members [the arginine (R) converted to histidine (H)]. In this pedigree, three out of eight family members were diagnosed as HCM and with a penetrance of 75%. In FHL family, mutation G15391A was identified in exon 23 of MYH7 gene in 3 family members [the glutamic acid (E) converted to lysine (K)]. In this pedigree, three out of six family members were diagnosed as HCM and with a penetrance of 100%. Echocardiography showed obstruction of left ventricular outflow tract in two out of the three HCM patients. CONCLUSIONS: Our results showed that the G12101A mutation of MYBPC3 gene is the causal mutation of familial HCM with mild phenotype. The G15391A mutation of MYH7 gene is the causal mutation of familial HCM with malignant phenotype and a penetrance of 100%. Screening mutations in the MYH7 gene should be viewed as a reasonable procedure in obstructive HCM patients. PMID- 19134270 TI - [Cardiac troponin I gene mutation (Asp127Tyr) in a Chinese patient with hypertrophic cardiomyopathy]. AB - OBJECTIVE: To observe the disease-causing gene mutation in Chinese patients with hypertrophic cardiomyopathy and to analyze the correlation between the genotype and the phenotype. METHODS: Specimens of peripheral blood were collected and the genome DNA was extracted in 65 unrelated patients with hypertrophic cardiomyopathy and 60 normal controls. The exon 7 and 8 of cardiac troponin I gene were screened with PCR and direct sequencing technique. RESULTS: A missense mutation in the exon 7 of the cardiac troponin I gene was identified in a 40-year old male patient with hypertrophic cardiomyopathy (Asp127Tyr) which was absent in the controls. CONCLUSION: A novel missense mutation of cardiac troponin I was identified in a patient with hypertrophic cardiomyopathy, this mutation might be the disease-causing gene mutation in this Chinese patient. PMID- 19134271 TI - [Titin gene mutations in Chinese patients with dilated cardiomyopathy]. AB - OBJECTIVE: Titin is recently known as the largest protein which exists in the striated muscle sarcomere and is dynamic both in biomechanics properties and biochemical functions. Four possible disease-associated mutations located in three exons (3, 14, 49) of titin gene (TTN) have been identified in Japanese DCM patients in 2002. We observed the possible association of TTN mutation in Chinese patients with DCM. METHODS: Three exons of TTN (3, 14, 49) were screened in 117 DCM patients and 120 controls by polymerase chain reaction-single strand conformation polymorphisms (PCR-SSCP) and DNA sequence. SSCP was carried out following a protocol optimized for each PCR fragment after amplification. Abnormal SSCP results were subsequently confirmed by DNA sequencing. RESULTS: The mutations reported in Japanese DCM patients were not identified in this patient cohort. A novel mutation [the G13053A (TTN cDNA sequence, X90568) change resulted in amino acid change at position 4351 (Gly4351Asp)] was found in two young DCM patients from a DCM family (1.7%). There was no similar mutation in controls. CONCLUSION: This novel Gly4351Asp mutation in TTN might be associated with DCM. PMID- 19134272 TI - [Imbalance of T helper 1 cells/T helper 2 cells accelerated T-cell-mediated endothelium injury in patients with acute coronary syndromes]. AB - OBJECTIVE: To observe the T helper 1 and T helper 2 (Th1/Th2) balance and possible association to vascular endothelial cells injury in patients with acute coronary syndromes (ACS). METHODS: Forty patients with ACS and 18 patients with stable angina pectoris (SAP) were included in this study. The concentrations of T helper 1/T helper 2 subsets related cytokines in plasma were evaluated by ELISA Kits. Cytotoxic activity of peripheral blood mononuclear cells (PBMCs) or PBMCs depleted CD(+) T cells against human umbilical vein endothelial cells (HUVECs) were evaluated by Cr51 cytotoxicity assay. RESULTS: Concentrations of T helper 1 related cytokines IFN-gamma and IL-2 were significantly higher [IFN-gamma: (131.2 +/- 42.2) ng/L vs. (47.6 +/- 20.2) ng/L; IL-2: (83.7 +/- 21.3) ng/L vs. (46.2 +/- 16.7) ng/L, all P < 0.05] while T helper 2 related cytokine IL-10 concentration was significantly lower [(16.7 +/- 4.3) ng/L vs. (27.5 +/- 5.5) ng/L, P < 0.05] in patients with ACS compared to those in SAP patients. Cytotoxic activity of PBMCs against HUVECs in patients with ACS was also significantly higher than that in patients with SAP (28.84% +/- 4.20% vs. 20.28% +/- 2.71%, P < 0.05). CONCLUSIONS: In patients with ACS, Th1 related cytokines were significantly upregulated while Th2 related cytokines were significantly downregulated. This imbalance of Th1/Th2 accelerated PBMCs mediated endothelium injury in patients with ACS. PMID- 19134273 TI - [Outcome of patients implanted with bioabsorbable polymer sirolimus-eluting stent (Excel) and the durable polymer sirolimus-eluting stent (Cypher)]. AB - OBJECTIVE: To compare the outcome of patients with coronary artery disease implanted with bioabsorbable polymer sirolimus-eluting stent (Excel) and the durable polymer sirolimus-eluting stent (Cypher). METHODS: One year clinical and 7 months angiographic follow-up data were analyzed in 591 consecutive patients implanted with Excel (E group: n = 350, 58.1 +/- 10.9 years old) or Cypher (C group: n = 241, 56.4 +/- 10.7 years old) during May 2006 to November 2006 in our center. RESULTS: There were more patients with old myocardial infarction and triple vessel diseases while there were less left main stem disease in E group compared to those of C group at baseline. Reference vessel diameter [(2.76 +/- 0.45) mm vs. (2.87 +/- 0.38) mm, P = 0.043], stent diameter [(3.11 +/- 0.48) mm vs. (3.25 +/- 0.34) mm, P = 0.045] and minimal stent diameter [(2.67 +/- 0.43) mm vs. (2.78 +/- 0.35) mm, P = 0.046] and rate of post-dilation (15.9% vs. 46.7%, P < 0.001) were all significantly smaller in E group compared to those in C group. Procedure was successful in all patients. One year clinical follow-up results showed similar results on major adverse cardiac events (MACE) rate (8.3% vs. 7.1%, P = 0.641), rate of cardiac death (0.9% vs.0.0%, P = 0.274), non-fatal myocardial infarction (2.0% vs. 1.2%, P = 0.747) and target vessel revascularization (5.4% vs. 5.8%, P = 0.857) between E and C groups. Total (0.9% vs. 0.4%, P = 0.649), early (0.6% vs. 0.0%, P = 0.516) and late (0.3% vs. 0.4%, P = 1.000) stent thrombosis rates according to Academic Research Consortium (ARC) standard definitions (definite + probable) were also similar between the E and C groups. Seven months angiographic follow-up results indicated that there were no differences of in-stent and in-segment restenosis rate between E and C groups (4.7% vs. 3.2%, P = 0.725; 5.6% vs. 3.2%, P = 0.505) but in-stent and in-segment late loss were significantly smaller in E group than those in C group [(0.18 +/- 0.07) mm vs. (0.21 +/- 0.08) mm, P < 0.001; (0.21 +/- 0.09) mm vs. (0.23 +/- 0.11) mm, P = 0.003]. CONCLUSION: One year clinical and 7 months angiographic outcome were comparable in patients implanted with Excel and Cypher sirolimus eluting stents. PMID- 19134274 TI - [Impact of patient compliance on the outcomes in hypertensive patients receiving hydrochlorothiazide based combination therapy with spironolactone or captopril]. AB - OBJECTIVE: To explore the impact of patient compliance on the long-term outcomes in hypertensive patients receiving hydrochlorothiazide (HCTZ) based combination therapy with spironolactone or captopril. METHODS: A total of 853 patients with mild to moderate hypertension were recruited and randomly divided into HCTZ group (HCTZ 12.5 mg q.d), spironolactone group (HCTZ 12.5 mg q.d and spironolactone 20 mg q.d), and captopril group (HCTZ 12.5 mg q.d and captopril 25 mg bid) after 2 week placebo washout period and 6-week loading period for HCTZ. Since the efficacy of combination therapy was proven to be better than monotherapy 1 year after therapy beginning, patients in HCTZ group were randomly assigned to spironolactone group or captopril group. The patients were followed up for 4 years. Patients were divided to compliance (n = 424) or non-compliance group (n = 429) according test drug taking questionnaire. During the follow-up time, the blood pressure and the outcomes were recorded monthly, and blood biochemical parameters were determined once a year. RESULTS: At the end of follow up, incidence of cardio-cerebral vascular events was significantly lower in compliance group (2 fatal, 8 non-fatal) than that in noncompliance group (7 fatal, 21 non-fatal, P < 0.05). Systolic blood pressure [-(19.4 +/- 20.6) mm Hg, 1 mm Hg = 0.133 kPa] and diastolic blood pressure [-(10.7 +/- 13.5) mm Hg] were significantly reduced compared values at baseline and noncompliance group (all P < 0.001) while the reduction did not reach statistically significance in noncompliance group [-(7.3 +/- 18.2) mm Hg and -(3.5 +/- 10.2) mm Hg, all P > 0.05 vs. baseline]. The serum BUN, Cr and UA levels in the compliance group were significantly higher and the serum K(+), CHO, LDL-C level were significantly lower than baseline values. The serum BUN, UA levels in the compliance group were significantly higher while the serum K(+), cholesterol levels were significantly lower than those in the noncompliance group (all P < 0.05). CONCLUSIONS: This study indicates that patient compliance could affect the long-term outcome and antihypertensive efficacy in hypertensive patients receiving HCTZ based combination therapy with spironolactone or captopril. PMID- 19134275 TI - [Electrocardiographic characteristics and radiofrequency catheter ablation results of ventricular tachycardia or premature ventricular contractions originating in the vicinity of atrioventricular annulus]. AB - OBJECTIVE: To evaluate the electrocardiographic (ECG) characteristics and results of radiofrequency catheter ablation (RFCA) of idiopathic ventricular tachycardia or premature ventricular contractions (VT/PVCs) originating in the vicinity of atrioventricular annulus. METHODS: Nineteen patients with idiopathic VT/PVCs underwent conventional electrophysiological study and RFCA were included in this analysis. The 12 leads (ECG) characteristics were also analyzed. RESULTS: The VT/PVCs were originated in the vicinity of mitral annulus in 10 cases, including anterolateral (n = 5), posterolateral (n = 3) and posteroseptal (n = 2). The VT/PVCs were originated in the vicinity of tricuspid annulus in the rest 9 cases, including the free wall (n = 5) and the septal portion (n = 4). The 12-lead ECG patterns of VT/PVCs originating in the vicinity of atrioventricular annulus were helpful for determine the site of RFCA. The VT/PVCs were successfully eliminated by RFCA in all cases. CONCLUSION: The exact origin of VT/PVCs originating in the vicinity of atrioventricular annulus could be determined by 12-leads ECG analysis and can be successfully and safely cured by RFCA. PMID- 19134276 TI - [Effects of intracoronary autologous bone marrow mononuclear cells transplantation in patients with dilated cardiomyopathy]. AB - OBJECTIVE: To evaluate the safety and efficacy of intracoronary autologous bone marrow mononuclear cells (BM-MNCs) transplantation in patients with dilated cardiomyopathy (DCM). METHODS: On top of standard therapy, DCM patients received BM-MNCs transplantation (n = 71) or saline injection (n = 187). The baseline clinical characteristics of two groups were comparable. Data on echocardiography, Holter, six-minute-walk test, cardiac SPECT and annual hospital days were obtained in all patients at baseline, 1, 3, 6, 12 and 24 months after transplantation. RESULTS: Six-minute-walk distance was significantly longer at one month [(345 +/- 76) m vs. (286 +/- 104) m, P < 0.05] and thereafter (all P < 0.05) in BM-MNCs group compared with saline group. Left ventri ocular ejection fraction (LVEF) at one month in BM-MNCs group was significantly higher compared with saline group [(41.5 +/- 9.4)% vs. (37.3 +/- 6.6)%, P < 0.05] and with pre transplantation value [(41.5 +/- 9.4)% vs. (32.4 +/- 8.5)%, P < 0.05] while LVEF was similar at 24 months after transplantation between the two groups [(43.6 +/- 6.3)% vs. (43.2 +/- 6.0)%, P > 0.05]. Three months after transplantation, the number of ischemic segments of BM-MNCs group was significantly reduced compared with that of saline group (2.0 +/- 1.0 vs. 3.1 +/- 1.4, P < 0.05) and with baseline (2.0 +/- 1.0 vs. 3.1 +/- 1.2, P < 0.05) while the number of necrotic segments were similar in both groups during the follow-up. There were no significant difference in survival between two groups during 2 years follow-up (95.4% vs. 94.9%, P > 0.05) but the annual hospitalization days of BM-MNCs group was significantly lower than that of saline group [(23.6 +/- 13.4) d vs. (33.0 +/ 14.0) d, P > 0.05]. CONCLUSIONS: Intracoronary transplantation of autologous BM MNCs was safe and could increase LVEF and the six-minute-walk distance and reduce hospitalization days for patients with dilated cardiomyopathy. PMID- 19134277 TI - [Incidence and risk factors of acute kidney injury post off-pump and on-pump coronary artery bypass grafting]. AB - OBJECTIVE: To investigate the incidence and risk factors of acute kidney injury (AKI) within the first postoperative week after off-pump or on-pump coronary artery bypass (OPCAB or CCAB) surgery. METHODS: Consecutive patients underwent CABG between January 1990 and August 2006 in our institution and had normal serum creatinine (Scr) and estimated creatinine clearance (Ccr) values before operation were retrospectively analyzed. Multivariate logistic regression analysis was performed to identify risk factors for the development of AKI defined as Scr 130 199 micromol/L or Ccr 30 - 60 mlxmin(-1)x1.73 m(-2). RESULTS: Incidence of AKI was significantly higher in patients underwent CCAB compared to those underwent OPCAB (63/331 vs. 61/518, P < 0.01). Peak Scr value was seen at 12th hour post OPCAB and 24th hour post CCAB, respectively. The rapid recovering of Scr occurred between 24th hour to 48th hour in patients underwent OPCAB and 48th hour to 72th hour in patients underwent CCAB surgery. Multivariate forward stepwise logistic regression analysis showed that LVEF < 30%, pulse pressure >/= 60 mm Hg (1 mm Hg = 0.133 kPa), peripheral vascular disease, diabetes, emergent procedure, triple vessel disease, higher body mass index (kg/m(2)), intraoperative and postoperative IABP, NYHA class III or IV and cardiopulmonary bypass were risk factors for the development of postoperative AKI following CABG, while LVEF > 50% and intraoperative and postoperative IABP were associated with lower incidence of AKI (OR < 1). CONCLUSION: AKI is not a rare complication post OPCAB or CCAB surgery, especially in patients with reduced LVEF, increased pulse pressure, peripheral vascular disease, diabetes, emergent procedure, triple-vessel disease, higher body mass index, intraoperative and postoperative IABP. PMID- 19134278 TI - [Effects of rhBNP on left ventricular remodeling in rats with acute myocardial infarction]. AB - OBJECTIVE: To assess the effects of rhBNP on left ventricular (LV) remodeling in rats with acute myocardial infarction (AMI). METHODS: AMI was induced by ligating coronary artery in male Sprague Dawley rats. Two days after surgery, AMI rats received intravenous infusion of rhBNP (15 microg/kg or 5 microg/kg once daily, n = 15 each) or saline (placebo control, n = 15) through Jugular Vein. Sham operated rats (n = 15) served as normal control. Four weeks later, hemodynamic measurements were performed, left ventricular weight (LVW), ratio of left ventricular weight to body weight (LVW/BW), left ventricular diameter (LVD) and infarct size were determined. Plasma angiotensin II and myocardial angiotensin II levels were also measured. RESULTS: Compared with sham-operated rats, LVW, LVW/BW, LVD and myocardial angiotensin II level were significantly increased, while the LV systolic pressure (LVSP), +/- dp/dt were significantly reduced in saline treated AMI rats (all P < 0.05). LVW/BW, MI size, LVD and myocardial angiotensin II in rhBNP treated AMI rats were significantly lower [LVW: (492.6 +/ 34.0) mg, (498.8 +/- 47.8) mg, (570.0 +/- 24.2) mg, P < 0.01; LVW/BW: 2.0 +/- 0.2, 2.0 +/- 0.2, 2.3 +/- 0.1, P < 0.01; LVD: (25.3 +/- 2.9)%, (31.4 +/- 3.0)%, (46.4 +/- 3.0)%, P < 0.01; myocardial angiotensin II: (881.3 +/- 62.7) pg/L, (1186.0 +/- 94.5) pg/L, (2436.7 +/- 280.3) pg/L, P < 0.05], while LVSP and +/- dp/dt in rhBNP treatment groups were significantly increased than saline treated AMI rats (P < 0.05 or P < 0.01). CONCLUSION: RhBNP is effective in attenuating left ventricular remodeling after AMI in rats. PMID- 19134279 TI - [Increased myocardial expression of calcium-sensing receptor and apoptosis in a rat model of atherosclerosis]. AB - OBJECTIVE: To observe the effect of hyperlipidemia and atherosclerosis on rat myocardial expression of calcium-sensing receptor and apoptosis. METHODS: The rat atherosclerosis model was induced by intraperitoneal injection of VD(3) (6 x 10(5) U/kg) and high cholesterol diet. Wistar rats were divided into two groups: (1) Control group; (2) AS group (n = 12 each). The expressions of CaSR, Bcl-2, Bax and caspase-3 were analyzed by Western blot and RT-PCR. Apoptotic cells were observed by TUNEL assay. The morphological changes of abdominal aorta and cardiac tissues were observed under optical and electro microscopes. The activity of LDH, CK, SOD and the content of MDA were assayed with ultraviolet spectrophotometer. The level of cTnT was detected by electrochemical immunofluorescence. RESULTS: Compared with control group, the activity of LDH and CK, the content of MDA and cTnT, the apoptosis index, the expression of CaSR, Bax and caspase-3 were significantly increased, but the SOD activity and Bcl-2 expression were significantly decreased, the myocardial ultrastructure injury was significantly aggravated in the AS group (all P < 0.05). CONCLUSION: Hyperlipidemia and atherosclerosis can up-regulate myocardial calcium-sensing receptor expression, promote myocardial apoptosis, aggravate oxidative stress and myocardial ischemia. PMID- 19134280 TI - [The effect of oxidized low-density lipoprotein on the maturation and its immune function of monocyte-derived dendritic cells]. AB - OBJECTIVE: To observe the effects of ox-LDL on monocyte-derived dendritic cells. METHODS: DCs were derived from healthy donors and divided into four groups according to the method of stimulation. The cells of blank group, negative group, experimental group and positive group which were treated with PBS, LDL, ox-LDL, TNF-alpha, individually. ox-LDL was added during the late stage of monocyte differentiation. Flow cytometry was used to analyze the cell surface markers and the endocytoses of DCs. (3)H-TdR incorporation was used to measure the proliferation of syngeneic and allogeneic T cells. ELISA assay was used to measure IL-12, MCP-1and MIP1 in cultured medium. Western blot analysis was used to evaluate the content of IkappaBalpha and NF-kappaB of DCs. RESULTS: Addition of ox-LDL during the late stage of monocytes differentiation can upregulate the cell surface markers including CD40 (22.3% vs. 45.6%) and CD86 (25.9% vs. 82.4%), increase the secretion of IL-12 (31.43 pg/ml vs. 126.73 pg/ml) and MCP-1 (59.6 ng/ml vs. 116.3 ng/ml), reduce DCs uptake capacity (46.8% vs. 10.7%), enhance allogeneic T cells proliferation (SI: 4.5 vs. 5.7), promote IkappaBalpha degradation and upregulate the expression of NF-kappaB in DCs. CONCLUSION: ox-LDL can promote the maturation of PBMCs-derived DCs by promoting IkappaBalpha degradation. PMID- 19134281 TI - [In vivo transfection of hepatocyte growth factor gene induces pulmonary angiogenesis in rabbits with hyperkinetic pulmonary hypertension]. AB - OBJECTIVE: To investigate the effect of adenoviral-mediated exogenous HGF (Ad HGF) gene transfer on lung angiogenesis in the rabbit lung in rabbits with hyperkinetic pulmonary artery hypertension. METHODS: A thoracotomy was performed through a midsternal incision in 1-month-old immature rabbit and an anastomosis between the left innominate artery and the pulmonary trunk was made to establish a chronic patent left to right shunt. Three months later, animals were randomly assigned to receive either Ad-HGF (2 x 10(9) Pfu in 0.2 ml PBS, H1 group), repeated administration of Ad-HGF after one week (H 2 group), Ad-GFP (2 x 10(9) Pfu in 0.2 ml PBS, G group), or PBS (0.2 ml, C group) by the intratracheal method of gene transfection. After two weeks, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical examination was performed to identify HGF mRNA and HGF protein expression. The capillary density and small pulmonary artery density were determined by immunostained with antibodies against factor VIII and alpha-SMA. After 1 month, the collateral vessels were evaluated by angiogram under digital subtraction angiography (DSA). RESULTS: Two weeks after Ad-HGF transfection, 484 bp bands could be found by RT-PCR in H1 and H2 groups, but not in other groups. The expression of HGF protein could be detected on alveolar epithelium and pulmonary vessel endothelium by immunohistochemistry examination. The number of factor VIII-positive pulmonary capillaries was also significantly increased in the H1 and H2 groups compared with the C and G groups (P < 0.05). The capillary density reached (17.0 +/- 3.3) mm(2) and (19.7 +/- 2.8) mm(2) in the H1 and H2 group, respectively, whereas it remained (13.2 +/- 3.2) mm(2) in the C group and (13.5 +/- 2.4) mm(2) in the G group (P < 0.05). One month after Ad-HGF transfection, the number of small pulmonary arteries was significantly increased in H1 and H2 group compared with control groups (P < 0.05). The collateral vessels were more abundant in HGF transfection groups than that in the two control groups reviewed by angiogram under digital subtraction angiography (DSA). CONCLUSION: In vivo gene transfection with HGF by means of the intra-tracheal injection could induce pulmonary angiogenesis in the early stage and small pulmonary arterial angiogenesis in later stage. PMID- 19134282 TI - [The effect of bone marrow mesenchymal stem cell transplantation on diabetic cardiomyopathy]. AB - OBJECTIVE: Diabetic cardiomyopathy (DCM), an important cause of heart failure, is characterized by microvascular pathologies and interstitial fibrosis. Bone marrow mesenchymal stem cells (MSCs) are pluripotent, which can differentiate into cardiomyocytes and vascular endothelial cells. They also secrete angiogenic and antiapoptotic factors. However, little information is available about the effect of MSCs transplantation on diabetic heart. METHODS: MSCs were isolated from bone marrow of isogenic adult rats and cultured ex vivo. Eight weeks post streptozotocin injection, saline or exogenous MSCs labelled with 4'6-Diamidino-2 Phenylindole (DAPI) were injected into the femoral vein of diabetic rats and examined 4 weeks later by echocardiography, histopathologic analysis, reverse transcription polymerase chain reaction analysis for matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1, zymography analysis for activities of MMP-2 and Western blot analysis for troponin T. RESULTS: Left ventricular posterior wall thickness and myocardial arteriolar density as well as the TIMP-1 mRNA and MMP-2 activity were significantly decreased in DCM group (P < 0.01 versus control group respectively), these changes were significantly attenuated by MSCs transplantation (P < 0.05 versus DCM). MSCs transplantation also significantly reduced fibrosis and downregulated MMP-9 mRNA in diabetic myocardium. CONCLUSION: Intravenous MSCs transplantation could attenuate LV remodeling in DCM rats. PMID- 19134284 TI - [Comparison of low density lipoprotein cholesterol levels in Kazaks and Mongolians from Xinjiang pastoral area]. AB - OBJECTIVE: To compare the serum levels of low density lipoprotein cholesterol in Kazaks and Mongolians from Xinjiang pastoral area. METHODS: A randomized cluster sampling was performed to obtain a representative sample of population in Xinjiang Hefeng pastoral area. Blood was taken from 632 individuals aged 30 and older for measurements of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) by automatic biochemical analyzer. The serum levels of low density lipoprotein cholesterol (LDL-C) was calculated according to the formula. RESULTS: The mean serum level of LDL-C was significantly higher in Kazaks [(3.68 +/- 1.16) mmol/L] than that in Mongolians [(3.29 +/- 1.23) mmol/L, P < 0.001]. The serum levels of LDL-C in Kazaks were significantly associated with mean artery pressure. The serum levels of LDL-C in Mongolians were significantly associated with body mass index. CONCLUSION: The serum levels of LDL-C were higher in Kazaks compared that in Mongolians in Xinjiang pastoral area. PMID- 19134283 TI - [Profile and associations of carotid femoral pulse wave velocity in a community based Beijing population of middle and old age]. AB - OBJECTIVE: To explore the profile and related factors of carotid femoral pulse wave velocity (CFPWV) in a community-based Beijing population of middle and old age. METHODS: A total of 1519 residents aged 44 - 79 years old from Shijingshan district, Beijing, were recruited for the cardiovascular risk factor survey and CFPWV measurements. One way ANOVA, pearson correlation and partial correlation, stepwise multi-linear regression were used for the data analysis. RESULTS: (1) The age-adjusted mean value of CFPWV was significantly higher in men than that in women (11.4 vs. 11.1 m/s, P = 0.007). CFPWV increased significantly with age both for men and women (all P < 0.01). (2) After adjusting for age and gender, CFPWV was significantly higher in people with hypertension, diabetes and higher heart rate (> 70 beats/min) than people without these factors (P < 0.01). (3) CFPWV significantly correlated with age, systolic blood pressure, diastolic blood pressure, fasting blood glucose and heart rate (correlation coefficient ranged from 0.12 - 0.46, all P < 0.01). In female, CFPWV was positively correlated with total cholesterol and triglycerides (correlation coefficient 0.11 and 0.13, all P < 0.01). Multi-linear regression analysis showed age, gender, systolic blood pressure, diastolic blood pressure, fasting blood glucose and heart rate were independent predictors of CFPWV. CONCLUSION: In this middle and old age Chinese population, age, sex, systolic blood pressure, diastolic blood pressure, fasting blood glucose and heart rate but not HDL-cholesterol and body mass index were associated with CFPWV. PMID- 19134285 TI - [Report of three cases transcatheter closure of trauma-induced muscular ventricular septal defects]. PMID- 19134286 TI - [Report of two cases extracorporeal membrane oxygenation therapy for patients with severe myocarditis]. PMID- 19134288 TI - [Vascular progenitor cells and atherosclerosis]. PMID- 19134291 TI - [Improving recognition of vascular cognitive impairment]. PMID- 19134289 TI - [Importance of adipo-vascular axis: from basic research to the clinic]. PMID- 19134292 TI - [The rise of neuropsychiatry: a breakthrough of the brain and mind barrier]. PMID- 19134294 TI - [Pay attention to depression of patients in department of neurology]. PMID- 19134295 TI - [Insights into invasive fungal infections in critical illness: interactions between guidelines and practice]. PMID- 19134296 TI - [Prevention of stroke relies on valid control "H" type hypertension]. PMID- 19134297 TI - [The role of training in internal medicine for the up-bringing and professional development of clinical doctors]. PMID- 19134298 TI - [The prevalence and risk factors for cognitive impairment following ischemic stroke]. AB - OBJECTIVES: To identify the prevalence and risk factors that were associated with post-stroke cognitive impairment (PSCI) among a large cohort of consecutive ischemic stroke patients. METHODS: 526 consecutive patients, who had suffered from ischemic stroke 3 months ago were recruited in this study. Patients were classified as having no cognitive impairment (NCI), cognitive impairment but no dementia (CIND) and vascular dementia (VaD) according to their cognitive function. They were also categorized as with subcortical ischemic vascular diseases (SIVD) or cortical ischemic vascular diseases (CIVD) with neuroimaging findings. Their demographic data, vascular risk factors and stroke characteristics were also documented. RESULTS: The overall prevalence of PSCI (CIND + VaD) was 36.7%. Compared with the NCI subjects, PSCI subjects were older, more dominant femininely, less educated, with more cases of right hemi-paralysis and higher depression scores, but did not have more specific vascular risk factor. Separately, VaD patients demonstrated lower economic level, less spouse caring, more prevalence of dysphasia, higher rate of incontinence and more cases with CIVD, while CIND patients had more cases with SIVD. The VaD patients had more cortical lesions and lower depression scores than the CIND patients. On logistic regression analysis, older age, female gender, lower economic level, dysphasia, SIVD, CIVD and higher depression scores were independent risk factors for PSCI. CONCLUSION: PSCI is common among ischemic stroke patients and related to demographic factors, stroke types, and depression. PMID- 19134299 TI - [Analysis of the changes in aortic root dimension in patients with anterior cerebral infarction]. AB - OBJECTIVE: To analyze the relationship between aortic root dimension (ARD) and anterior circulation infarction (ACI) by measuring ARD of ACI patients. METHODS: The ARD of 110 ACI patients (56 men and 54 women) were measured with 2 dimensional ultrasonography during ventricular systole. 66 control subjects (32 men and 34 women) without the clinical signs of cerebral infarction or organic heart disease and cardiac insufficiency were selected after carrying out detailed case history inquiring, physical examination, CT or MRI and ultrasonic examination. RESULTS: ARD in men and women was respectively (29.80 +/- 2.35) mm and (28.88 +/- 3.06) mm in the ACI group. While it was (27.64 +/- 1.70) mm and (26.98 +/- 1.60) mm respectively in the control subjects. ARD in men and women was significantly higher in the ACI group as compared with the control subjects (P < 0.01). Logistic regression analysis indicated that the odds ratio (confidence interval) associated with ARD was 1.630 (1.205 - 2.204) in men and 1.494 (1.098 - 2.033) in women for ACI. CONCLUSIONS: There is a close relationship between ARD and ACI. ARD is an important risk factor of ACI. PMID- 19134300 TI - [A prospective study for systemic inflammatory response syndrome (SIRS) after cerebral infarction]. AB - OBJECTIVE: To investigate whether systemic inflammatory response syndrome (SIRS) after cerebral infarction is associated with poor outcome and its associated clinical factors. METHODS: We prospectively studied 500 patients with cerebral infarction, recorded the associated clinical factors on presentation and calculated the mortality at 21st day. RESULTS: There are 85 SIRS patients in 500 patients with cerebral infarction, 31 with total anterior cerebral infarction (TACI), 34 with partial anterior cerebral infarction (PACI), 15 with posterior cerebral infarction (POCI) and 5 with lacunar cerebral infarction (LACI). The frequency of SIRS was positively correlated with mortality rate according to Oxfordshire Community Stroke Project (OCSP) subtypes in cerebral infarction patients with fever (Spearman correlation coefficient = 1.0; P < 0.001). Single factor analysis showed that the risk factors for both SIRS and mortality of cerebral infarction were as follows: age, infection, 48 h Chinese Stroke Scale score, 48 h Glasgow score, OCSP subtypes and dysphagia. Diabetes was the risk factor for SIRS, but had no effect on mortality. On Cox regression, 48 h Glasgow score was the sole independent risk factor of outcome. However, if SIRS was included in the formula, P > 0.05. CONCLUSIONS: SIRS is the predictor of poor outcome after acute cerebral infarction. It is important to prevent SIRS especially in TACI and POCI. Diabetes is the risk factor for SIRS, but has no effect on mortality. PMID- 19134301 TI - [A clinical research of hirayama disease]. AB - OBJECTIVE: To summarize and analyze the clinical features of Hirayama disease. METHODS: The authors summarized the clinical data of 29 cases of Hirayama disease in recent 9 years. RESULTS: All of the 29 cases were male and the age of onset was 12 - 25 years. 22 of the 29 patients only had the symptoms of one upper limb and 7 of them had symptoms of both upper limbs. Most of them had symptoms on the right or the symptoms were on more severe in the right than left, while only one case had symmetric symptoms of both upper limbs. The arm appeared as a slope, this was due to partial muscular atrophy. EMG indicated that the impairment of spinal anterior cells were limited to the arm relevant segments and both sides were involved. Cervical MRI scan suggested spinal atrophy at the level of C(5 - 7) vertebral bodies, being asymmetric in most of the cases. The atrophy was more obvious on the side with more severe symptoms. There was one patient whose MRI showed high T(2) signal in the spinal anterior cells of the C(5 - 6) segments. All the cases used cervical supporter and were followed up for 1 - 5 years. All of them had good prognosis. CONCLUSIONS: Hirayama disease is more common in men with hidden onset before 20 years of age. Most of the patients noted the symptoms of hands gradually about 2 years after a period of rapid growth in height in adolescence. The predominant clinical manifestations are as follows: atrophy of lower cervical cord in cervical MRI, asymmetric spinal atrophy in horizontal impaction and the degree of atrophy consistent with the symptoms of limbs. EMG could indicate bilateral spinal anterior impairment in lower cervical cord. Rare case may present the abnormal signal in the spinal cord, but pyramid signs were absent. PMID- 19134303 TI - [An association between glucose metabolism status and brachial-ankle pulse wave velocity in Chinese patients with coronary artery disease]. AB - OBJECTIVE: To investigate the association between glucose metabolism status and brachial-ankle pulse wave velocity (baPWV) in Chinese patients with coronary artery disease (CAD). METHODS: baPWV values were measured in 198 CAD patients. Plasma glucose, serum lipids, hsCRP and other baseline data were obtained in all the patients. According to the glucose level, the participants were divided into 3 groups: normal glucose tolerance (NGT) group, impaired glucose regulation (IGR) group and diabetes mellitus (DM) group. RESULTS: baPWV values in the DM group (1807 +/- 381) cm/s were significantly higher than those in the NGT group [(1615 +/- 248) cm/s, P = 0.000] and IGR group [(1674 +/- 277) cm/s, P = 0.035]. Multiple stepwise linear regression analyses indicated that higher baPWV values were associated with aging and hsCRP levels in 198 patients. In DM group, higher baPWV values were independently associated with aging and HbA1c levels. CONCLUSIONS: In patients with coronary artery disease, baPWV values increase with different glucose metabolism status from NGT to IGR then DM. baPWV values in the patients with DM were significantly higher than those in the other two groups. Glycemic control may ameliorate arterial stiffness. PMID- 19134304 TI - [Analysis of clinical features of autoimmune disease-related pancreatitis]. AB - OBJECTIVE: To improve the understanding of autoimmune disease related pancreatitis by analyzing their clinical features. METHODS: The clinical features were analyzed retrospectively in 28 autoimmune disease related pancreatitis cases from Peking Union Medical College Hospital (PUMCH), according to the associated autoimmune diseases. RESULTS: (1) The average age was (40.0 +/- 16.1) years, and the ratio of male to female patients was 1:6. There were 24 acute and 4 chronic pancreatitis in the 28 cases. (2) The common related autoimmune diseases were systemic lupus erythematosus (20/28) and Sjogren's syndrome (6/28). (3) The characteristics of the autoimmune diseases was multi-system involvement, such as hematologic system, kidney, liver, etc. (4) Clinical features of those acute pancreatitis shown that no distinct trigger exist for acute pancreatitis, and the radiological changing was not prominent. (5) In laboratory examination, an obvious increase of CA199 could be seen, paralleling the severity of pancreatitis. (6) Glucocorticoids or immunosuppressors was effective, and the mortality rate of acute pancreatitis cases was 33.3%. CONCLUSIONS: Autoimmune disease related pancreatitis is dominant with acute pancreatitis and females is common, which may reflect the activity of autoimmune diseases. Autoimmune disease related acute pancreatitis has a high mortality rate. Glucocorticoids and/or immunosuppressors may be useful to relieve the pancreatitis. PMID- 19134305 TI - [A clinical study on gestational transient thyrotoxicosis]. AB - OBJECTIVE: To investigate the prevalence of gestational transient thyrotoxicosis (GTT) and analyze the cause of thyrotoxicosis encountered in this period. METHODS: An epidemiologic survey in ten hospitals in Shenyang was performed and 534 pregnant women during the first trimester of pregnancy filled questionnaire, received physical examination and had serum thyroid-stimulating hormone (TSH), free T(4) (FT(4)), free T(3) (FT(3)), thyroid peroxidase antibody (TPOAb), thyrotrophin receptor antibody (TRAb), and human chorionic gonadotrophin (hCG) tests. RESULTS: (1) The total prevalence of thyrotoxicosis was 9.75% (52/534) in the first trimester and the prevalence of GTT was 7.86%, which accounted for 80.77% of the thyrotoxicosis encountered in this period. A total of 88.89% of the overt GTT showed only elevated FT(3) level. (2) The level of serum hCG increased gradually in the first trimester. The medians of hCG were 25 300, 85 220 and 81 780 IU/L 6, 8 - 10 and 12 weeks after gestation, respectively (P = 0.000). The medians of serum TSH were 1.45, 1.10 and 0.84 mIU/L 6, 8 - 10 and 12 weeks after gestation, respectively (P < 0.01). (3) When serum hCG was more than 50 000 IU/L, the prevalence of GTT increased obviously. When serum hCG was between 80 000 IU/L and 110 000 IU/L, subclinical GTT increased significantly. When serum hCG was more than 110 000 IU/L, overt GTT increased significantly. Correlation analysis showed that serum hCG was related negatively with TSH (r = -0.402, P = 0.000) and positively with FT(3) (r = 0.165, P = 0.000), but not related with FT(4). CONCLUSIONS: The prevalence of GTT is 7.86% in the first trimester and it is the main cause of thyrotoxicosis found in the first trimester, accounting for 80.77% of all the causes. The serological characteristic of overt GTT is mainly the elevation of serum FT(3) level. Serum hCG level is related with the severity of GTT. PMID- 19134306 TI - [Predictors of maternal and fetal outcome in systemic lupus erythematosus: a retrospective study of 94 cases]. AB - OBJECTIVE: To evaluate the predictors of maternal and fetal outcome of pregnancy for systemic lupus erythematosus (SLE) patients. METHODS: Ninety-four patients with 96 pregnancies which were evaluated retrospectively from Jan 1990 to Jan 2008 in Peking Union Medical College Hospital were divided into two groups: disease stable during pregnancy (group A) and lupus flares during pregnancy (group B). Statistical analysis was performed by chi(2) or Fisher exact test and Student's t-test. A binary logistic regression model was used to evaluate the predictors of maternal and fetal outcome. RESULTS: There were 36 pregnancies with stable lupus disease (group A) and 60 pregnancies with lupus flares (group B). Of the 96 pregnancies, 18 resulted in therapeutic abortion and 7 in fetal loss, 71 resulted in a live birth,3 in neonatal death. The rates of preterm delivery, small gestational age (SGA) and neonatal asphyxia in group B were higher than those in group A (P < 0.05). By binary logistic regression analysis, preeclampsia/eclampsia low serum platelet count and SLE flares were associated with poor fetal outcome (beta = 2.463, 2.228, 2.769 respectively, P < 0.05). There were 56 pregnancies with stable lupus disease at the conception with 22 (39.3%) occurred lupus flares during pregnancies. Twenty-four preeclampsia and 2 eclampsia were seen in all the pregnancies. Fifty-two pregnancies were complicated with lupus nephritis, and 25 pregnancies (48.1%, 25/52) of which were disease stable at the conception, and among 22 pregnancies with disease stable over one year, twelve of which occurred lupus nephritis flares. Three pregnancies which have disease activity within one year before pregnancy all occurred lupus nephritis flares. There were four maternal death which all occurred at the postpartum. By binary logistic regression analysis, lupus nephritis flares were associated with preeclampsia/eclampsia (beta = 2.658, P < 0.05), and proteinuria at the conception before delivery were significantly associated with SLE flares (beta = 3.263, P < 0.05). CONCLUSION: An increase of fetal loss, preterm delivery, SGA and neonatal asphyxia was seen in patients with lupus flares during pregnancy compared with those with stable disease. About 1/3 lupus activity may increase after pregnancy. Preeclampsia and eclampsia were increased when there were lupus nephritis flares. PMID- 19134307 TI - [Clinical efficacy and safety of itraconazole injection/oral solution sequential therapy for treatment of invasive fungal infection in intensive care unit]. AB - OBJECTIVE: To investigate the therapeutic efficacy and safety of itraconazole injection/oral sequential therapy on invasive fungal infection (IFI) in ICU. METHODS: In this multicenter, post-marketing, open-label study, ICU patients who have met the inclusion IFI criteria will be enrolled in this study. Itraconazole intravenous injection is administered 200 mg twice a day in day 1 - 2, then 200 mg once a day at least for 5 days, and maintenance itraconazole oral solution as sequential therapy, itraconazole oral solution 200 mg twice a day sequential therapy lasts for 2 weeks. Clinical efficacy and adverse reaction were record. RESULTS: A total of 159 patients were enrolled and completed this trial. (1) At the end of first week, total clinical cure rate was 35.2%, and increased to 73.6% after the second week. Clinical cure rate were 72.9% and 72.2% in possible and probable IFI patients, and 78.9% in proven IFI patients at the end of second week. (2) At the end of first week, total fungal clearance was 40.9%, and increased to 75.9% and 92.9% at the end of second and fourth week. Fungal clearance were 90.0% and 64.6% in possible and probable IFI patients, and 84.2% in proven IFI patients at the end of second week respectively. (3) Combined clinical cure rate and fungal clearance, the total clinic efficacy was 44.1% at the end of first week, and increased to 92.9% and 100.0% at the end of second and fourth week. (4) No severe adverse reaction was found. CONCLUSIONS: Itraconazole injection/oral sequential therapy is an effective and safe antibiotic for the treatment of IFI in ICU. PMID- 19134308 TI - [A clinical study of 49 cases of invasive pulmonary aspergillosis]. AB - OBJECTIVE: Studying the proven and probable invasive pulmonary aspergillosis (IPA) cases of some hospitals in Shanghai to provide evidence for the improvement of IPA clinical diagnosis and therapy. METHODS: Forty-nine IPA cases were retrospectively analyzed for demography data, host factors, underlying conditions, chest CT, microorganism and histopathology examination, as well as therapy and clinical outcome. RESULTS: Of 49 subjects including 19 (38.8%) proven and 30 (61.2%) probable IPA, 3 patients (6.1%) had no host factors, 25 patients (51.0%) had IPA associated host factors and underlying conditions, while 21 patients (42.9%) had uncertain fundamental diseases. Chest CT evaluation demonstrated that radiological lesions include nodules in 29 patients, patching in 15, mass in 12, consolidation in 10, cavitation in 34, Halo sign in 19, air bronchogram in 18, crescentic sign in 6, bilateral in 33 and multifocal lesions in 38. The yielding rate of fungus culture in sputum was 26.5% (13/49), and in bronchoalveolar lavage fluid was 66.7% (10/15). Eleven of thirty-six patients (30.6%) had positive results of serum galactomannan antigen tests. Nineteen of twenty-one patients (90.5%) were proven as IPA by lung histologic examinations. Aspergillus fumigatus was the most common pathogen 81.0% (17/21). The responding rate to initial anti-fungus therapy was 50% (21/42). CONCLUSION: Our study suggests that in IPA patients, bilateral, multifocal and nodular lesion could be the most common radiological characteristic, while Halo and crescentic sign occur occasionally. Invasive technologies are more valuable to IPA diagnosis. PMID- 19134309 TI - [A retrospective study of efficacy and safety of itraconazole for treatment of invasive fungal infection in hematologic diseases]. AB - OBJECTIVE: To investigate the efficacy and safety of intravenous itraconazole for the treatment of invasive fungal infection (IFI) in patients with hematological disease or undergoing allogeneic hematopoietic stem cell transplantation (HSCT). METHODS: Six hundred and sixty-six patients with above mentioned conditions and diagnosed as IFI from January, 2007 to July, 2007 were enrolled. Intravenous itraconazole was administered at a dose of 200 mg every 12 hours for 2 days and followed by 200 mg every 24 hours. Patients were then switched to oral itraconazole according to the clinical situation. Responses were determined on the basis of clinical and microbiological criteria. RESULTS: The probability of defervescence was 69.8% and the total response rates among related to itraconazole were the proven, probable and possible IFI patients were 73.7%, 68.1% and 68.2% (P = 0.380). Adverse effects were found in 58 patients (8.7%), which were mainly mild to medium reversible dysfunction of liver and gastrointestinal tract. CONCLUSION: Itraconazole is an effective and safe antifungal agent for patients with hematological disease or undergoing HSCT and is suitable for empirical antifungal therapy. PMID- 19134310 TI - [Sequential intravenous and oral itraconazole in treatment of invasive fungal infection in hematological diseases]. AB - OBJECTIVE: We performed this study to investigate the efficacy and safety of sequential intravenous and oral itraconazole in the treatment of invasive fungal infection (IFI) in the patients with hematological diseases. METHODS: All the patients fit for the diagnostic criteria of IFI (proven diagnosis, probable diagnosis and possible diagnosis) with hematological diseases were entered into this study. The trial was designed as an open labeled, multi-centered study, with the total treatment duration of 4 - 6 weeks including intravenous and oral period. Intravenous itraconazole was given at the dose of 200 mg, twice a day in the first two days, and adjusted to 200 mg/d for subsequent 12 days. Sequential oral itraconazole was given at the dose of 200 mg, twice a day, for 2 - 4 weeks for maintenance therapy. Efficacy and safety evaluation was performed every 1 - 2 weeks according to the physician's decision. RESULTS: Two hundred and twenty seven patients were enrolled into this study. The overall response rate was 75.33% and the cure rate was 47.14%. Two hundred and five patients (205/227, 90.3%) defervesced after a median of 5 days (2 - 20 days). One hundred and eighty six patients were applicable for microbiological evaluation, among them, 69.89% responded to the treatment. Eleven treatment-related adverse events occurred during the study and none of them were severe adverse events. CONCLUSION: Sequential intravenous and oral itraconazole therapy was effective for IFI and tolerated well in patients with hematological diseases. PMID- 19134322 TI - [Current situation and prospect of radiological vascular interventional therapy in obstetrics and gynecology]. PMID- 19134323 TI - [The relevance between the effect on dysmenorrhea and the time of surgery of artery embolization in the treatment of adenomyosis]. AB - OBJECTIVE: To analyze the relevance between the effect on dysmenorrhea of uterine artery embolization (UAE) in treatment of adenomyosis (AM) in different periods of the menstrual cycle and the time of the operation. METHODS: Totally 225 cases with preoperative dysmenorrhea voluntarily chose UAE in treatment of AM. They were divided into two groups according to the different times of menstrual cycle (proliferative phase or secretory phase). We analyzed the relevance between the effect of dysmenorrhea of patients in the two groups with the time of surgery after 1, 2, 3 and 4 years. RESULTS: (1) The numbers of the cases completely followed up were 142, 128, 119 and 101 each year in the following four years. (2) The effective rate on dysmenorrhea in the two groups of AM patients was 81% (43/53) and 76% (68/89) after 1 year (P > 0.05). (3) The effective rate was 75% (36/48) and 70% (56/80) after 2 years (P > 0.05). (4) The effective rate was 70% (32/46) and 63% (46/73) after 3 years (P > 0.05). (5) The effective rate was 63% (24/38) and 63% (40/63) after 4 years (P > 0.05). CONCLUSION: UAE has a good efficacy in treatment of AM, but the relevance is not significant between the time of surgery and the effectiveness on dysmenorrhea. PMID- 19134325 TI - [Monitoring of methotrexate concentrations in lung and other tissues of rat through internal iliac artery infusion]. AB - OBJECTIVE: To establish the high-performance liquid chromatography (HPLC) method for measuring concentrations of methotrexate (MTX) in rat lung and some other tissues through internal iliac artery infusion. METHODS: Fifty female Sprague Dawley rats were included in this study. The rats were randomly assigned to two groups. Methotrexate was injected to group one through internal iliac artery, and was injected to group two through femoral vein. Blood and tissues were collected in each group at 15, 30, 60, 90 and 120 minutes for detection of the drug concentrations with HPLC. RESULTS: The area under the concentration time curve (AUC) in rat lung, ovary and uterus in the artery group were separately (3.77 +/- 0.28), (4.40 +/- 0.40), (9.97 +/- 0.89) microgxh(-1)xg(-1), which were significantly different from those of the vein group [(2.31 +/- 0.25), (3.91 +/- 0.19), (7.65 +/- 1.54) microgxh(-1)xg(-1); P < 0.05]. The AUC in the rat plasma, heart, kidney, liver and spleen in the artery group were separately (6.13 +/- 0.53), (1.90 +/- 0.11), (5.32 +/- 0.89), (14.16 +/- 1.96), (0.76 +/- 0.20) microgxh(-1)xg(-1). There were no significant differences from the vein group [(5.79 +/- 0.71), (1.64 +/- 0.29), (5.15 +/- 1.69), (14.29 +/- 3.47), (0.76 +/- 0.13) microgxh(-1)xg(-1); P > 0.05]. CONCLUSIONS: Through internal iliac artery infusion, there are higher drug concentrations in lung, uterus and ovarian compared to venous injection. The internal-arterial chemotherapy may be used to treat pulmonary metastasis of gynecological tumor. PMID- 19134324 TI - [Comparison of effectiveness between intra-arterial and intra-venous neoadjuvant chemotherapy in stage Ib2-IIb cervical carcinoma]. AB - OBJECTIVE: To compare the effect between intra-arterial and intra-venous neoadjuvant chemotherapy (NACT) in stage Ib2-IIb cervical carcinoma. METHODS: A retrospective analysis was done on 52 cases of intra-venous NACT and 95 cases of intra-arterial NACT for stage Ib2-IIb cervical carcinoma treated in Peking Union Medical College Hospital from 1999. RESULTS: The response rate of intra-venous NACT and intra-arterial NACT was 88% (46/52) and 79% (75/95), and the operative rate after NACT was 81% (42/52) and 72% (68/95) respectively (P > 0.05). There were no significant differences in surgery time, blood loss and post-operative morbidity between these two groups. Pathological parametrial positive rate after NACT in arterial group (6%) was significantly lower than that of venous group (50%, P > 0.05). The venous group had very similar recurrence rates (13% vs 17%) and death rates (9% vs 12%) when compared with the arterial group (P > 0.05). CONCLUSIONS: The intra-arterial and intra-venous NACT for stage Ib2-IIb cervical carcinoma show similar response rate, operative rate and surgical difficulties. Arterial NACT shows a better effect on parametrial infiltration. PMID- 19134326 TI - [Etiology of primary ovarian insufficiency and premature ovarian failure]. PMID- 19134327 TI - [Role of CD4+ CD25 high regulatory T cells in the pathogenesis of intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the role of CD(4)(+)CD(25)(+) regulatory T cells and CD(4)(+)CD(25)(high) regulatory T cells in peripheral blood and decidua in the pathogenesis of intrahepatic cholestasis of pregnancy (ICP). METHODS: Peripheral blood and decidua CD(4)(+)CD(25)(+) regulatory T cells and CD(4)(+)CD(25)(high) regulatory T cells in 30 ICP patients (15 mild ICP and 15 severe ICP) and 28 normal pregnant women were analyzed by flow cytometry. RESULTS: The percentage of CD(4)(+)CD(25)(+) regulatory T cells and CD(4)(+)CD(25)(high) regulatory T cells in the CD(4)(+) T cells population in both peripheral blood and decidua in ICP patients were significantly lower than those in control women [(7.96 +/- 1.32)% vs (17.05 +/- 2.86)%, (17.18 +/- 2.27)% vs (32.01 +/- 3.88)%; (0.78 +/- 0.22)% vs (1.71 +/- 0.69)%, (2.25 +/- 0.89)% vs (8.30 +/- 1.13)%; P < 0.01]. Meanwhile, a significantly higher percentage of CD(4)(+)CD(25)(+) regulatory T cells and CD(4)(+)CD(25)(high) regulatory T cells in the CD(4)(+) T cells population was observed in all decidua samples compared to peripheral blood both in ICP patients (P < 0.01) and in control women (P < 0.01). In addition, the percentages of CD(4)(+)CD(25)(+) regulatory T cells and CD(4)(+)CD(25)(high) regulatory T cells in the CD(4)(+) T cell population in both decidua and peripheral blood in severe ICP [(15.94 +/- 1.95)%, (7.17 +/- 1.17)%, (1.87 +/- 0.90)%, (0.68 +/- 0.19)%] were lower than those in mild ICP [(18.43 +/- 1.90)%, (8.74 +/- 0.96)%, (2.62 +/- 0.72)%, (0.89 +/- 0.20)%] and normal pregnancy (all P < 0.05). CONCLUSIONS: CD(4)(+)CD(25)(high) regulatory T cell may play an important role in the pathogenesis of ICP and control of disease progression. PMID- 19134328 TI - [Expression and significance of chemokine CXC receptor 3, 4 and their ligands at the early pregnancy decidua and villi]. AB - OBJECTIVE: To explore the expression and significance of chemokine CXC receptor (CXCR) 3 and CXCR4 and their ligands (CXCL) at the early pregnancy decidua and villi. METHODS: Decidual mononuclear cells were isolated from the normal decidua of 5 - 8 weeks pregnant women by lymphocyte separation medium in vitro. CD(56)(+) natural killer (NK) cells were purified by dynabeads cell sorter kit. Purity and phenotype of CD(56)(+) decidua NK cells were analyzed by fluorescence-activated cell sorter (FACS). Gene expression of CXCR3 and CXCR4 in decidua NK cells and CXCL9, CXCL10 and CXCL12 in early pregnancy decidua and villi was assessed by RT PCR. Protein expression of CXCL9, CXCL10 in normal endometrium and early pregnancy decidua was characterized and quantified by streptavidin-biotin peroxidase chain reaction (SP) immunohistochemistry and computered image analysis system. Correlations between the gray degree of CXCL9 and CXCL10 and the number of CD(56)(+)NK cells in upper tissue were analyzed by Spearman's correlation coefficient rank test. RESULTS: The phenotype of 98.7% decidua NK cells was CD(56)(bright). The genes of CXCR3 and CXCR4 were expressed in decidua NK cells and that of CXCL9 and CXCL10 were expressed in early pregnancy decidua and CXCL12 in early pregnancy villi. CXCL9 and CXCL10 were expressed in the cytoplasm of surface epithelia, glandular epithelia and stromal cells of early pregnancy decidua and were not expressed in villi by immunohistochemistry. The gray degree of CXCL9 and CXCL10 in the secretory phase endometrium (56 +/- 43, 59 +/- 47) was stronger than that in the proliferative phase (16 +/- 18, 8 +/- 14, P < 0.05) and reached the highest (143 +/- 35, 158 +/- 29, P < 0.05) in the early pregnancy decidua. The number of CD(56)(+) NK cell in the secretory phase endometrium (60 +/- 20) was more than that in the proliferative phase endometrium (23 +/- 4, P < 0.05) and was the most in the early pregnancy decidua (114 +/- 15, P < 0.05). The gray degree of CXCL9 in upper tissue had a positive correlation with the number of CD(56)(+) cells (r = 0.88, P < 0.05) and that of CXCL10 had a similar pattern to CXCL9 (r = 0.86, P < 0.05). CONCLUSION: The interactions between CXCL9, CXCL10 and CXCL12 expressed in decidua and villi and CXCR3, CXCR4 expressed in CD(56)(+) decidua NK cells may influence the CD(56)(+)NK cell recruitment at the maternal fetal interface. PMID- 19134329 TI - [Correlation between killer cell immunoglobulin-like receptors genes and pre eclampsia]. AB - OBJECTIVE: To analyze the killer cell immunoglobulin-like receptors (KIR) gene polymorphism of pre-eclampsia patients and approach the correlation between KIR genes and pre-eclampsia. METHODS: The KIR gene polymorphisms of 71 pre-eclampsia patients and 100 healthy pregnant women were detected by PCR-SSP. The KIR2DL4 mRNA level in placentas from pre-eclampsia and gestational normal pregnancies were quantified by real time RT-PCR. Forty pre-eclampsia patients and 38 healthy pregnant women were detected for single nucleotide polymorphisms (SNP) in the gene coding and joint areas between introns and extrons by directly sequencing techniques of KIR2DL4 genomic DNA. Finally, all alleles and genotypes of KIR2DL4 gene were case-control studied. RESULT: Distributions of some relatively activating KIR genotypes shewed a significant association with pre-eclampsia. Real-time RT-PCR showed that KIR2DL4 mRNA can be measured both in placenta of women with pre-eclampsia being of pre-eclampsia was significantly lower than that of normal pregnancy, only as much as 0.276 times that of controls. We identified 18 polymorphisms, of which, 7 were first reported. But no significant differences in genotype distributions or allele frequencies were observed in these SNPs between cases and controls. CONCLUSION: The distributions of some relatively activating KIR genotypes showed a significant association with pre-eclampsia, which indicates that the polymorphism of KIR genes may be associated with the genetic predisposition to pre-eclampsia. And because the expression of KIR2DL4 mRNA in the placentas of cases was significantly lower than control group, it is speculated that the decrease of KIR2DL4 expression in placenta may participate in the pathogenesis of pre-eclampsia. PMID- 19134330 TI - [Clinical significance of hypoxia inducible factor-prolyl hydroxylase 1 and factor inhibiting hypoxia inducible factor-1 expression in placentas of women with severe pre-eclampsia]. AB - OBJECTIVE: To investigate the role of hypoxia inducible factor (HIF)-prolyl hydroxylase 1 (HPH1) and factor inhibiting HIF-1 (FIH-1) in placentas in the pathogenesis and development of severe pre-eclampsia. METHODS: RT-PCR and western blot analyses were used to detect the HPH1 and FIH-1 expression levels in placentas of 34 patients with severe pre-eclampsia and 24 cases of term pregnancy (normal pregnancy group) and their correlations with symptoms were analyzed. RESULTS: (1) The HPH1 mRNA and protein expression levels in placentas of severe pre-eclampsia group were 0.40 +/- 0.04 and 59.5 +/- 3.4 separately, significantly lower than those of normal pregnancy group, 0.84 +/- 0.12 and 71.6 +/- 1.7 (P < 0.01). The FIH-1 mRNA and protein expression levels in placentas of severe pre eclampsia group were 0.31 +/- 0.05 and 45.6 +/- 2.4 separately, significantly lower than those of normal pregnancy group, 0.43 +/- 0.04 and 54.9 +/- 2.1 (P < 0.01). (2) The mRNA and protein expression levels of HPH1 and FIH-1 in severe pre eclampsia group were all negatively correlated with mean arterial pressure (MAP) [the Spearman correlation coefficient was -0.854 (P < 0.01)], urinary protein per 24 hours [the Spearman correlation coefficient was -0.936 (P < 0.01)] and the occurrence of fundus oculi artery spasm [the Spearman correlation coefficient was -0.854 (P < 0.01)]. (3) The expression of HPH1 mRNA in placentas of all the 58 cases was 0.58 +/- 0.27, higher than the expression of FIH-1 mRNA, which was 0.39 +/- 0.10. There was a positive correlation between them. The pearson correlation coefficient was 0.686 (P < 0.01). The expression of HPH1 protein in placentas of all the 58 cases was 64.5 +/- 6.7, higher than the expression of FIH-1, which was 49.4 +/- 5.2. There was a positive correlation between them. The Pearson correlation coefficient was 0.947 (P < 0.01). CONCLUSION: The expression imbalance of HPH1 and FIH-1 in placenta may play an important role in the pathogenesis and development of severe pre-eclampsia through inhibiting HIF 1alpha. PMID- 19134331 TI - [Influence of 4-aminopyridine on human ovarian luteinized granulosa cell proliferation, production, and apoptosis through inhibiting voltage-gated K+ channel]. AB - OBJECTIVE: To study the influence of 4-aminopyridine (4-AP) on proliferation, production, and apoptosis through inhibiting voltage-gated K(+) channel (Kv) in ovarian luteinized granulosa cells. METHODS: Ovarian luteinized granulosa cells were recovered from 25 women with regular menses who underwent in vitro fertilization programme. The cultured granulosa cells were divided into 4 groups:blank group, 4-AP treated group, human chorionic gonadotropin (hCG) induced group and hCG + 4-AP co-treated group. The final concentrations of hCG and 4-AP were 1250 U/L and 5 nmol/L respectively. The progesterone production was detected by the chemoluminescence method. The expression of Kv mRNA on human ovarian luteinized granulosa cell was detected by RT-PCR. The influence on the early apoptosis of granulosa cells by 4-AP was observed by flow cytometry. Cellular caspase-3 activities were observed with colorimetric method and the inhibition of the cell proliferation was studied using methyl thiazolyl tetrazolium (MTT) method. RESULTS: (1) Kv mRNA was expressed in granulosa cell. (2) The progesterone production of the blank group, 4-AP treated group, hCG induced group and hCG + 4-AP co-treated group were (547 +/- 64), (206 +/- 32), (1991 +/- 172) and (763 +/- 79) nmol/L, respectively after 24 hours culture. Exposure of the granulosa cells to 4-AP reduced the production of progesterone in blank and hCG-induced granulosa cells. (3) The flow cytometry analysis and the cellular caspase-3 A(405) showed that 4-AP increased the percentage of early phase apoptosis (P < 0.01): 4-AP treated group vs blank group [(40 +/- 5)% and 0.049 +/- 0.009] vs [(17 +/- 4)% and 0.029 +/- 0.008], hCG + 4-AP co-treated group vs hCG-induced group [(25 +/- 4)% and 0.039 +/- 0.008] vs [(15 +/- 3)% and 0.022 +/- 0.007]. (4) 24 hours after treated with 4-AP and hCG, the inhibitory rate of cultured granulosa cells of 4-AP treated group was higher than the blank group (19.7% vs 0), and that of hCG + 4-AP co-treated group was obviously higher than hCG-induced group (34.6% vs 0, P < 0.01). CONCLUSIONS: The voltage-gated K(+) channels expressed by ovarian luteinized granulosa cell play an important role in cell proliferation, production, and apoptosis. 4-AP may inhibit differentiation of progesterone in granulosa cells through the inhibition of proliferation and induction of apoptosis. PMID- 19134332 TI - [Clinical analysis of 42 cases of primary malignant tumor in vagina]. AB - OBJECTIVE: To analyze the clinical characters, treatment and prognosis of primary malignant tumor in vagina. METHODS: A retrospective analysis of 42 patients diagnosed with primary malignant tumor in vagina in Peking Union Medical College Hospital (PUMCH) between Jan 1984 and Aug 2006 was performed. RESULTS: Primary malignant tumor accounted for 0.98% (42/4286) in the total gynecological malignant tumors during that period in PUMCH. According to the International Federation of Gynecology and Obstetrics (FIGO) staging system, 19 cases were at stage I, 12 cases at stage II, 5 cases at stage III, and 6 cases at stage IV. Thirteen cases were squamous carcinoma, 13 cases were malignant melanoma, 8 cases were adenocarcinoma, 3 cases were yolk sac tumor and 5 cases were other types. The majority of patients were treated with surgery combined with radiotherapy and chemotherapy. Up to August 2007, 19 cases survived, 18 cases were dead and 5 cases were lost. The longest follow up was 10 years, with the median time of 2 years. The overall 2-year survival rate was 60.6%. For stage I, stage II and stage III - IV, the 2-year survival rates were 71.3%, 58.3% and 29.6% respectively. The 2-year survival rate of patients with squamous carcinoma was 46.8%, malignant melanoma 72.9%, adenocarcinoma 20.0% and patients with yolk sac tumor were all alive tumor-free after 6 - 10 years' follow up. CONCLUSIONS: The prognosis of primary malignant tumor in vagina is affected by clinical stage and histological type. As to malignant melanoma, radical surgery combined with chemotherapy and immunotherapy produce good effects. Patients with yolk sac tumor can be cured only with chemotherapy. As to other types, more treatment experiences are needed. PMID- 19134333 TI - [Pulmonary resection in the management of gestational trophoblastic neoplasia: a clinical study]. AB - OBJECTIVE: To investigate the effect of surgical resection in the management of gestational trophoblastic neoplasia (GTN) patients with pulmonary metastases. METHODS: A retrospective review of the medical records of 62 GTN patients who underwent pulmonary resection was carried out. The cases were divided into recurrent group (group A, n = 10), drug-resistant group (group B, n = 28), and the group with satisfactory response to chemotherapy but residual pulmonary lesion (s) (group C, n = 25). One patient underwent lobectomy twice, and she was allocated simultaneously to groups A and B. The patients' median age, antecedent pregnancy, International Federation of Gynecology and Obstetrics (FIGO) risk score, number of preoperative chemotherapy courses, preoperative beta-human chorionic gonadotrophin (beta-hCG) titer, lesion size, number of lobes affected, positive rate of histology, follow-ups and prognosis were compared between the three groups. RESULTS: The proportion of high-risk patients in the three groups was 90%, 82% and 44%, respectively. The complete remission rates of the three groups were 90%, 79% and 100%, with relapse rates of 2/8, 15% and zero, respectively. Positive histology of the resected specimen was more frequently recognized in recurrent and drug-resistant groups (A 60%, B 36%, C 12%). In the drug-resistant group there were more preoperative chemotherapy sessions (A 3, B 7, C 5) and more patients with abnormal preoperative beta-hCG titer (A 50%, B 61%, C 12%). CONCLUSIONS: Surgical resection is effective in the treatment of pulmonary metastases of GTN. Surgery is indicated when clinical evidence suggests that pulmonary metastatic disease causes relapse or drug-resistance and the lesions are relatively localized. Surgical resection is not recommended for patients with satisfactory response to chemotherapy but residual pulmonary lesions. PMID- 19134334 TI - [The values of serum human epididymis secretory protein 4 and CA(125) assay in the diagnosis of ovarian malignancy]. AB - OBJECTIVE: To evaluate the value of human epididymis secretory protein 4 (HE4) and CA(125) in the diagnosis of ovarian malignancy. METHODS: HE4 and CA(125) in the serum specimens of malignant ovarian tumor group (30 cases), benign ovarian diseases (110 cases; 45 benign ovarian tumor, 57 endometriotic diseases and 8 pelvic inflammation were included) and healthy women group (137 cases) were assayed double blindly. The levels and the diagnosis efficiency of the HE4 and CA(125) were analyzed. RESULTS: (1) The median levels of HE4 and CA(125) were significantly higher in malignant ovarian tumor group (244 pmol/L and 601 kU/L respectively) than those of the benign ovarian diseases group (32 pmol/L and 22 kU/L respectively) and healthy women group (32 pmol/L and 11 kU/L respectively) (P = 0.000 - 0.029). The median levels of CA(125) were also higher in endometriotic diseases and pelvic inflammation groups (53 and 41 kU/L respectively) than those of benign ovarian tumor group and healthy women group (12 and 11 kU/L respectively; P = 0.000 - 0.031). (2) The positive rate of HE4 was lower than that of CA(125) in malignant ovarian tumor group (P = 0.036). HE4 was negative in benign diseases and healthy women groups. But the positive rates of CA(125) were 56.1% and 5/8 respectively in endometriotic diseases and pelvic inflammation groups and there were significant differences compared with HE4 (P = 0.000). (3) The HE4 assay had advantage over the CA(125) assay in receiver operating characteristic-area under the curve (ROC-AUC) and sensitivity with a specificity of 100% when ovarian malignancy was compared with controls having benign diseases and healthy women, benign tumor or benign diseases groups respectively. The CA(125) assay had advantage over the HE4 assay in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having healthy women group. (4) Combined assay of HE4 and CA(125) was better than CA(125) alone when ovarian malignancy was compared with controls having any group. (5) Combined assay was better than HE4 alone in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having benign diseases and healthy women or healthy women groups. And combined assay was lower in the ROC-AUC and the sensitivity with specificity of 100% than HE4 when ovarian cancers were compared with controls having benign tumors or benign diseases groups respectively. (6) The diagnosis efficiency of the HE4 assay at the level 86 pmol/L determined in ROC curve with controls having benign diseases and healthy women group and at the 95% reference level 50 pmol/L of healthy women or 150 pmol/L recommended by the kit respectively was compared. The sensitivity of 50 pmol/L was 73% higher than 150 pmol/L and 86 pmol/L, while the specificity and positive predictive value were lower (P = 0.002, P = 0.000). The specificity, accuracy and positive predictive value of HE4 assay at the set point of 150 pmol/L and 86 pmol/L were 100%, 96% and 96%. The set point of 86 pmol/L had advantage over 150 pmol/L at the sensitivity of diagnosis, 70% and 63% respectively. But the positive predictive value was 95% lower than 150 pmol/L, being 100%. There was no significant difference (P = 0.883, P = 0.883). CONCLUSIONS: The specificity of HE4 assay is higher than CA(125) assay in the diagnosis of ovarian cancer and HE4 combined with CA(125) assay can improve the diagnoses. The set point of 150 pmol/L is advantageous for the accurate diagnosis, while the set point of 86 pmol/L is advantageous for the screening of malignant ovarian cancer. PMID- 19134335 TI - [Effect of estradiol supplementation during the luteal phase on mouse endometrial expression of leukaemia inhibitory factor and pinopodes in controlled ovarian stimulation cycles]. AB - OBJECTIVE: To study the effect of estradiol supplementation during the luteal phase on mouse endometrial expression of leukaemia inhibitory factor and pinopodes in controlled ovarian stimulation cycles. METHODS: Female mice were randomly divided into four groups: group A [controlled ovarian stimulation (COS) group], group B (COS group with progesterone for luteal-phase-support), group C (COS group with progesterone and estradiol for luteal-phase-support), and group D of natural cycle group. Pinopodes were investigated by scanning electronic microscopy (SEM) in the uterine endometrium of pregnant mice on pregnancy days (pd) 3 - 5. Leukaemia inhibitory factor (LIF) protein was determined by immunohistochemistry in the uterine endometrium of pregnant mice on pd 3 - 5. RESULTS: (1) In groups B, C, and D, there were small developed pinopodes in the endometrial surface of pregnant mouse on day 3; there were large fully developed pinopodes in endometrial surface, which was smooth with well defined borders resembling a mushroom on day 4. The regressing pinopodes were observed on day 5. In group A, there were small developed pinopodes in endometrial surface of pregnant mouse on day 3. The regressing pinopodes were seen on day 4. (2) In the pregnant mice of groups C and D, the level of LIF protein on days 3 - 5 (138.5 +/ 20.3, 143.1 +/- 19.0) was significantly higher than group A (103.2 +/- 5.0, P < 0.05), and strong immunostaining of LIF protein was found on day 4 of gestation. In group B, the level of LIF protein on days 3 - 5 (123.5 +/- 10.8) was significantly higher than group A (P < 0.05), but significantly lower than groups C and D (P < 0.05). Strong immunostaining of LIF protein was found on day 4 of gestation. In group A, weak immunostaining of LIF protein peaked on day 3 of gestation. In groups B, C, and D, the level of LIF protein on day 4 was significantly higher than group A on day 3 (F = 55.76, P < 0.01). CONCLUSIONS: Estradiol supplementation during the luteal phase can improve the expression of LIF and pinopodes in mouse endometrium in controlled ovarian stimulation cycles and redress the harmful effect on implantation window by COS. Therefore, estradiol supplementation can improve the endometrial receptivity. PMID- 19134342 TI - [Clinical application of positron-emission tomography for the identification of cervical nodal metastases of head and neck cancer compared with CT or MRI and clinical palpation]. AB - OBJECTIVE: To evaluate the value of positron-emission tomography (PET) for the identification of cervical nodal metastases of head and neck cancer compared with CT/MRI and clinical palpation. METHODS: Forty patients of head and neck cancer underwent PET and CT/MRI examination 2 weeks before surgery. PET, CT/MRI and clinical palpation were interpreted separately to assess regional lymph node status. Histopathologic analysis was used as the gold standard for assessment of the lymph node involvement. Differences in sensitivity, specificity and accuracy among the imaging modalities and clinical palpation were analyzed. RESULTS: The sensitivity of PET for the identification of nodal metastases was 14.3% higher than that of CT/MRI (P = 0.648) and 14.3% higher than that of clinical palpation (P = 0.648), whereas the specificity of PET was 15.4% higher than that of CT/MRI (P = 0.188) and 7.7% higher than that of clinical palpation (P = 0.482). The accuracy of 18F-FDG PET, CT/MRI, and clinical palpation for the identification of cervical nodal metastases was 85.0%, 70.0% and 75.0% respectively. CONCLUSIONS: The sensitivity, specificity and accuracy of PET for the detection of cervical nodal metastases was higher than that of CT/MRI and clinical palpation. Although the results did not show a statistically significant difference, PET can still serve as a supplementary method for the identification of nodal metastases of head and neck cancer. PMID- 19134344 TI - [Radiation-induced maxillary malignancies: complications of radiotherapy treatment for head-neck malignant tumors]. AB - OBJECTIVE: To investigate the incidence of radiation-induced maxillary malignancy after radiotherapy for head and neck cancer. METHODS: A total of 273 patients who suffered from osteoradionecrosis after radiotherapy for head and neck cancer were evaluated. Among them, 6 patients were presented with carcinoma and sarcoma arising from maxillary area after radiotherapy. RESULTS: Radiation-induced maxillary cancers happened at a rate of 2.2% in the patients with osteoradionecrosis. There were no statistically significant differences in age, sex and the time interval between the radiotherapy and the cancer occurrence. CONCLUSIONS: Radiation-induced malignancy after radiotherapy for head and neck cancer is mainly located in maxilla, presenting as squamous cell carcinoma or sarcoma of the maxillary sinus. PMID- 19134343 TI - [Molecules of G(2)/M phase and the phosphorylation of survivin in the carcinogenesis of oral submucosal fibrosis]. AB - OBJECTIVE: To determine the expression of Cyclin B1, p34(cdc2) and the phosphorylation of survivin (p-survivin) in oral squamous cell carcinoma and oral submucosa fibrosis (OSF), and to discuss their possible role in carcinogenesis of OSF. METHODS: The expression of Cyclin B1, p34(cdc2) and p-survivin were analyzed by Western blotting assay in 10 cases of normal oral mucosa epithelium, 40 cases of OSF epithelium and 42 cases of oral squamous cell carcinoma (OSCC) originated from OSF, respectively. Immunoprecipitation was used to confirm the relationship between the p34(cdc2) and survivin. RESULTS: The expression of Cyclin B1, p34(cdc2), p-p34(cdc2) and p-survivin in OSF group were significantly higher than those in normal group (P < 0.05). The expression of these molecules showed significant different (P < 0.05) between the OSF and OSCC originated from OSF, but there was no significant difference among the early stage, the moderately advanced stage and the advanced stage of OSF. Immunoprecipitation assay confirmed the combination of p34(cdc2) and survivin. CONCLUSIONS: The important molecules in G(2)/M phase-Cyclin B1, p34(cdc2) and p-survivin may play a key role during the mitosis and proliferation of OSF, which will be helpful in early diagnosis and therapy of carcinogenesis of OSF. PMID- 19134345 TI - [Reconstruction of the tongue with lateral arm free flap after hemiglossectomy]. AB - OBJECTIVE: To investigate the value of microsurgical reconstruction of the tongue after hemiglossectomy with lateral arm free flaps in the treatment of tongue cancer. METHODS: From October 2006 to April 2007, ten patients underwent simultaneous reconstruction of the tongue and oral floor defects with lateral arm free flaps after resection of squamous cell carcinoma of tongue in our hospital. The flaps ranged from 8 cm x 5 cm to 9 cm x 6 cm in size, and were adjusted to the defect of the tongue. The vascular pedicle included the posterior radial collateral artery and the accompanying veins. The outcome of reconstruction was evaluated by follow-up examinations, including the contour and mobility of the reconstructed tongue, the swallowing function and the speech function. RESULTS: All patients recovered uneventfully from surgery, without any major postoperative complications. The transplanted flaps survived. During follow-up period the contour of the reconstructed tongues was satisfactory. The patients demonstrated good functional mobility of the reconstructed and remaining tongue. Postoperatively, the swallowing and speech function was nearly at normal levels and the patients could ingest a solid or semisolid diet. CONCLUSIONS: Microvascular reconstruction of the tongue with lateral arm free flaps after hemiglossectomy is an ideal and safe method, but long term results need to be further studied. PMID- 19134346 TI - [Preliminary clinical application of Tip-Edge Plus appliance in the treatment of Class II patients]. AB - OBJECTIVE: To evaluate the craniofacial and occlusional changes of Class II patients treated with Tip-Edge Plus technique. METHODS: Twelve Class II patients (7 girls and 5 boys, mean age 14.3 years) with bimaxillary or maxillary protrusion were selected. All the patients were four first premolar extraction cases and treated with Tip-Edge Plus technique. Cephalometric analysis was performed to evaluate the changes before and after treatment. RESULTS: After treatment, the profile was greatly improved. UL-E, LL-E were decreased significantly (P < 0.01) and incisors were retracted (P < 0.05). Upper molars moved forward slightly (P > 0.05). CONCLUSIONS: The anchorage control in Tip-Edge Plus technique was effective without extraoral force, palatal arch and implants. Under very light force, the crown of anterior teeth tipped distally and lingually quickly. In the final stage, it was possible to achieve both desired root uprighting and crown torque. PMID- 19134347 TI - [Orthodontic treatment of Class III patients with mandibular asymmetry]. AB - OBJECTIVE: To investigate the treatment outcome of Class III patients with dental, functional and mild skeletal mandibular asymmetry. METHODS: Thirty-five patients (14 males and 21 females) with dental, functional and mild skeletal mandibular asymmetry were selected. The age range of the patients was 7 - 22 years with a mean age of 16.5 years. Dental mandibular asymmetry was treated with expansion of maxillary arch to help the mandible returning to normal position. Functional mandibular asymmetry was treated with activator or asymmetrical protraction and Class III elastics. Mild skeletal mandibular asymmetry was treated with camouflage treatment. RESULTS: Good occlusal relationships were achieved and facial esthetics was greatly improved after orthodontic treatment in patients with dental and functional mandibular asymmetry. However, patients with skeletal mandibular asymmetry should be treated with both extraction and genioplasty. CONCLUSIONS: Orthodontic treatment was suitable for patients with dental and functional mandibular asymmetry, while combined orthodontics and surgery could get good results in patients with skeletal mandibular asymmetry. PMID- 19134348 TI - [Matrix metalloproteinases regulations of human gingival fibroblasts by Porphyromonas gingivalis with different fimA genotypes]. AB - OBJECTIVE: To investigate the mechanism of matrix metalloproteinases (MMP) regulations of human gingival fibroblasts (HGF) by challenge of Porphyromonas gingivalis (Pg) with different fimA genotypes. METHODS: Pg ATCC 33277 (type I), WCSP115 (type II), WCSP1.5 (type III), W83 (type IV) were assessed for their inductions of MMP-1 and MMP-2 expression in HGF. MMP mRNA levels of HGF were determined by real-time RT-PCR and MMP protein levels in culture supernatant were determined by ELISA at different time intervals (1, 3, 6 and 12 h) following continuous co-culture of bacteria with HGF. RESULTS: When co-cultured with Pg, the MMP-1 and MMP-2 mRNA and protein expression of HGF significantly increased compared with the negative control group (P < 0.01). The group of type II showed greater up-regulated than other fimA genotypes in the mRNA and protein expressions of MMP-1 and MMP-2, MMP-1 mRNA [(28.88 +/- 3.12) - (231.01 +/- 24.99)] and protein [(1.35 +/- 0.17) - (3.08 +/- 1.20)] microg/L; MMP-2 mRNA [(20.42 +/- 2.21) - (188.34 +/- 37.37)] and protein [(2.57 +/- 0.76) - (18.08 +/- 1.15)] microg/L for different time periods; While the group of type III was weaker than other fimA genotypes, the level of MMP-1 mRNA was [(5.11 +/- 0.55) - (72.84 +/- 8.84)] and protein [(0.68 +/- 0.13) - (1.46 +/- 0.94)] microg/L, MMP-2 mRNA [(4.55 +/- 0.55) - (25.75 +/- 3.12)] and protein [(2.28 +/- 0.93) - (11.22 +/- 2.46)] microg/L (P < 0.05). CONCLUSIONS: Pg could induce HGF to over-express MMP, and fimA genotypes of Pg may be related to this pathogenicity, which might indicate fimA genotype is associated with pathogenesis of Pg. PMID- 19134349 TI - [Effects of calcitriol on the expression of vitamin D receptor, RANKL and osteoprotegerin in human periodontal ligament cells]. AB - OBJECTIVE: To study the effects of 1, 25-dihydroxyvitamin D(3) (VD(3)) on the expression of vitamin D receptor (VDR), receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) in human periodontal ligament cells (hPDLC) populations and to analyze the potential mechanisms. METHODS: Twelve hPDLC populations were primarily established from 12 donors individually. Two samples of each hPDLC population of passage three were treated respectively with 10(-8) mol/L VD(3) (V D(3) group) or 0.1% absolute ethyl alcohol as controls (control group). Six days later, the mRNA expression levels of VDR, RANKL and OPG in the samples were determined with real-time quantitative RT-PCR. The DNA base sequences upstream to the transcription start site of RANKL gene were also analyzed. RESULTS: Compared with the control group, the mRNA expression level of VDR increased significantly in the VD(3) group (P = 0.003), averagely (3.04 +/- 1.06) times of that in the control group; the mRNA expression level of RANKL was also up-regulated by VD(3) (P = 0.001), 9.82 (0.75-119.18) times of that in the control group; the OPG expression level was (94.48 +/- 39.15)% of the controls (P = 0.136); OPG/RANKL ratio was down-regulated in the VD(3) group to averagely 10.36% (1.01%-138.00%) of the controls (P = 0.003). No mutation was found in the DNA fragments upstream to the transcription start site in the RANKL gene and the genotypes of the polymorphism at -1832 (rs7984870, C/G) were not shown to be significantly related to the RANKL mRNA expression level. CONCLUSIONS: In hPDLC, VD(3) can significantly increase the mRNA expression level of VDR; VD(3) can increase RANKL mRNA expression level to decrease OPG/RANKL ratio, but it has little effect on OPG mRNA expression. The big differences of the RANKL mRNA regulation in response to VD(3) treatment among hPDLC populations may not be associated with the DNA sequences upstream to the transcription start site in the RANKL gene. PMID- 19134350 TI - [Bacteriological analysis of subgingival plaque in adolescents]. AB - OBJECTIVE: To investigate the changes in characteristics of periodontal pathogens in subgingival plaque of patients with puberty gingivitis and its relevance with clinical symptoms. METHODS: A total of 108 subgingival plaque samples were collected from 30 patients with puberty gingivitis (Group G), 9 cases of chronic periodontitis (Group P) and 15 healthy controls (Group H). The age of the 54 participants was from 11 to 17. The black-pigmented bacteria (BPB), Fusobacterium nucleatum (Fn), Actinobacillus actinomycetemcomitans (Aa), Actinomyces were detected using bacterial culture. The probing depth (PD), gingival index (GI), bleeding index (BI) and attachment loss (AL) were also recorded. RESULTS: In all these three groups, the detection rates of black-pigmented bacteria were: 3%, 30% and 100%; Fn were: 30%, 68% and 94%, statistically significantly different (P < 0.01). The lgCFU/ml of black-pigmented bacteria and Actinomyces was higher in mild-moderate group [(3.8 +/- 0.7) and (5.3 +/- 0.9)] than in Group H (P < 0.001). The lgCFU/ml of black-pigmented bacteria and Fn significantly was higher in severe inflammation group [(4.7 +/- 1.2) and (4.4 +/- 0.8)] than in the mild moderate group (P < 0.01). The lgCFU/ml of black-pigmented bacteria, Fn and Aa was higher in severe gingivitis group [(6.6 +/- 1.0), (5.5 +/- 1.0) and (4.2 +/- 1.7)] than in mild gingivitis group (P < 0.01). The detection rate and lgCFU/ml of black-pigmented bacteria, Fn and Aa were both positively correlated with BI, PD and AL. CONCLUSIONS: In the stage of severe gingivitis, the periodontal pathogens increased markedly, suggesting that risk of further destruction of periodontal tissue may exist. PMID- 19134352 TI - [Stress distribution in alveolar bone around implants under implant supported overdenture with linear occlusion at lateral occlusion]. AB - OBJECTIVE: To analyze stress distribution in alveolar bone around implants of implant supported overdentures (ISO) with linear occlusion and with anatomic occlusion at lateral mandibular position, and to justify the possibility of decreased injurious force around implants in ISO with linear occlusion. METHODS: Computerized tomography scan and finite element analysis (FEA) were used to set up two 3-D FEA models of maxillae and mandible with severe residual ridge resorption. The mucosa, linear and anatomic occlusal ISO with bar attachments, and two implants inserted between mandibular foramina were also established in the models. With the condition of imitating the loading of masseter muscles, these models were loaded to simulate the stress distributions in alveolar bone around implants under ISO at lateral occlusion position. RESULTS: At lateral occlusion, the stress distributions in alveolar bone around implants under ISO with anatomic occlusion were mainly on the lingual and distal sides of the working side implants. However, stress distributions under ISO with linear occlusion were on the distal sides of bilateral implants. Both the stress peaks of ISOs with linear occlusion and with the anatomic one appeared in the working side. In anatomic occlusion model, sigma(z): -6.47 MPa and 6.81 MPa, sigma(1): 4.20 MPa and 7.20 MPa (negative value: compressive stress, positive value: tensile stress); in linear occlusion model, sigma(z): -4.86 MPa and 3.04 MPa, sigma(1): -3.48 MPa and 5.33 MPa. CONCLUSIONS: At lateral occlusion, when comparing the ISO with two different occlusion schemes, stress peak in alveolar bone around implants in the linear occlusion model was lower than that in the anatomic occlusion model at equal loading situation. Stress in the alveolar bone under ISO with linear occlusion distributed more evenly than that under ISO with anatomic occlusion. PMID- 19134351 TI - [A new ultrasonic system in initial periodontal therapy: clinical efficacy and perception of pain]. AB - OBJECTIVE: To compare the clinical efficacy and perception of pain during initial periodontal treatment using Vector-system and a conventional ultrasonic system. METHODS: Thirty-eight patients with mild to moderate periodontitis were debrided using both A-system (Vector-system) and B-system (Suprasson P5 Newtron) in a split mouth design. At baseline, one month after treatment, plaque index (PLI) and bleeding on probing (BOP) were recorded and probing depth(PD) was measured. A verbal analogue scale (VAS) was used to evaluate the perceived pain right after the treatment. RESULTS: At one month evaluation, all areas showed an improvement in clinical parameters. No statistically significant differences in PD and BOP could be observed between areas treated by two different systems, while A-system contributed to greater improvement in PLI than B-system. Patients treated with A system [(22.5 +/- 7.5) mm] experienced about 60% the amount of pain compared with the conventional ultrasonic device [(37.2 +/- 11.3) mm]. CONCLUSIONS: During initial periodontal treatment for the patients with mild to moderate periodontitis, A-system could achieve a comparable clinical efficacy and reduced pain sensations to conventional ultrasonic device. PMID- 19134353 TI - [Microstructure and mechanical properties of laser melted rapidly solidified forming pure titanium specimens]. AB - OBJECTIVE: To compare the microstructure and mechanical properties of pure titanium specimens fabricated using laser melted rapidly solidified forming (LMRSF) and casting, and to provide scientific evidence for the new method of laser rapid forming in prosthodontic field. METHODS: Six pure titanium specimens were fabricated from pure titanium powder using LMRSF technology, and six pure titanium casting specimens were used as the controls. The tensile strength was measured and the microstructures and natural fracture surface of the pure titanium specimens made in two different ways were observed. RESULTS: Compared to that in the casting specimen, the microstructure of the LMRSF titanium specimens was denser and acicular structure distributed more uniformly. The tensile strength and the microhardness of the pure titanium specimens were (510.0 +/- 21.2) MPa and (201.4 +/- 14.5) MPa respectively, which were much higher than those of the dental pure titanium castings [(425.0 +/- 35.1) MPa and (186.5 +/- 9.5) MPa] (P < 0.01). CONCLUSIONS: The microstructure and mechanical properties of the pure titanium specimens fabricated using LMRSF are better than that of the pure titanium castings. Therefore, the LMRSF will provide titanium framework with better quality for removable dental restoration. PMID- 19134354 TI - [An 8-year follow-up study of Cerec2 computer aided design and computer aided manufacture of all-ceramic crowns]. AB - OBJECTIVE: To evaluate the clinical performance of the crowns made by Cerec2 computer aided design and computer aided manufacture (CAD/CAM) system. METHODS: Twenty four teeth were restored with Cerec2 CAD/CAM crowns. Eight years after delivery of the prostheses, the all-ceramic crowns were evaluated using modified standard of California Dental Association quality evaluation system. Patients filled out a satisfaction questionnaire at the same time. And Kaplan-Meier analysis was carried out. RESULTS: Three out of 24 restorations were rated as failure. Kaplan-Meier analysis showed that the success rate of Cerec2 crowns eight year following delivery was 88%. CONCLUSIONS: Crowns made by Cerec2 CAD/CAM system can survive in oral condition and function well for 8 years. PMID- 19134355 TI - [Effect of hydrostatic pressure on ultrastructure of synovial fibroblasts from rat temporomandibular joint]. AB - OBJECTIVE: To investigate the role of apoptosis in the remodeling of temporomandibular joint (TMJ) under pressure. METHODS: Synovial fibroblasts obtained from rat temporomandibular joint were subjected to different hydrostatic pressure for 12 h. Changes of ultrastructure were observed by transmission electron microscope. RESULTS: At 30 kPa, the ultrastructure of synovial fibroblasts showed no obvious changes. At 60 kPa, the chromatin was condensated, the baryon took on crescent and the mitochondria seemed varicose. At 90 kPa, the apoptosis-like body was wrapped by membrane and embedded in the high density chromatin. CONCLUSIONS: Apoptosis-like change took place in ultrastructure of synovial fibroblasts under hydrostatic pressure, and the degree of the change was related to the hydrostatic pressure exerted. PMID- 19134356 TI - [Roles of matrix metalloproteinases and tissue specific inhibitors of metalloproteinases in dentinogenic ghost cell tumor and ghost cell odontogenic carcinoma]. AB - OBJECTIVE: To investigate the roles of matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in dentinogenic ghost cell tumor (DGCT) and ghost cell odontogenic carcinoma (GCOC). METHODS: The expressions of MMP-2, MMP-9, MMP-14, TIMP-1 and TIMP-2 were examined in 15 DGCT cases and 9 GCOC cases by immunohistochemistry. Their mRNA expression in one DGCT case and one GCOC case were investigated by RT-PCT.MMP-2 and MMP-9 protein activities in the two cases were analyzed by gelatin zymography. RESULTS: MMP-9 and TIMP-1 expressions elevated greatly in GCOC, and there was a significant difference (P < 0.05) in TIMP-1 expression between GCOC and DGCT.Pro-MMP-9, MMP-9 activated form, pro-MMP 2, and MMP-2 activated forms were detected in the GCOC case, while pro-MMP-9 and MMP-9 activated form were very faint in the DGCT case. The mRNA level of MMP-9 elevated obviously in the GCOC case, which was similar to that of TIMP-1. CONCLUSIONS: The elevated expression of MMP-9 and TIMP-1 may influence the behaviour of GCOC. PMID- 19134357 TI - [Rights maintenance and self-discipline of practicing dentist: Part XVI. Responsibility for quoting proof in lawsuits for medical care dissension]. PMID- 19134358 TI - [Chromatological knowledge and shade matching: Part III. Method of shade matching and impress of color message]. PMID- 19134359 TI - [Management of lymph nodes in level II(b) during selective neck dissection for clinically N(0) neck in oral and oropharyngeal cancer]. PMID- 19134360 TI - [Advancement of molecularly targeted therapies for salivary gland cancers]. PMID- 19134361 TI - [Rapid prototyping technology in the diagnosis and treatment of maxillary embedded teeth in the design of clinical application]. PMID- 19134362 TI - [The application of surgery-related disciplines to the medical relief work in earthquake disaster]. PMID- 19134363 TI - [Mass casualty critical care for surgical patients during a disaster]. PMID- 19134364 TI - [The idea of management for the injured patients caused by earthquake]. PMID- 19134367 TI - [Analysis of patients with bone injury in Wenchuan earthquake]. AB - OBJECTIVE: To evaluate the patients with bone injury in Wenchuan earthquake. METHODS: From May 12th to June 15th 2008 the data of 1410 patients with bone injury in Wenchuan earthquake were analyzed to evaluate clinical intervention and remedy-managing experience. RESULTS: The 1410 patients average age was from 4 to 103 years old. And 744 cases (52.7%) suffered from blunt injuries, 379 cases (26.9%) from buried injuries, 287 cases (20.4%) from falling injuries; And 1317 cases were with fracture, 93 with limbs soft tissue injuries; 261 patients combined with other parts of injuries including 45 cases with paralysis; 66 cases were with crush syndrome, 25 with gas gangrene, 76 with acute kidney failure, 26 with multiple organ failure. And 912 operations were performed including 402 fracture fixation, 224 debridement, 152 debridement and suture, 85 amputation, 29 implant skin, 8 fixation of joint dislocation, 5 surgical flaps transplantation, 4 nerve and tendon suture, 2 arthroscopes, 1 joint replacement. Among the 66 crush syndrome patients, 49 accepted continuous renal replacement therapy, in which 9 cases were bleeding from named arteries and 20 blood vessels were getting embolism. Among the 1410 cases, 1 died from multiple organ failure. CONCLUSIONS: Among the patients with bone injury in Wenchuan earthquake, the elderly patients are more than the youth; The injuries are always combined with other complications; Opened injuries are polluted severely; It is difficult to deal with the crush syndrome; Paraplegia cases are less, but the amputees are more. PMID- 19134368 TI - [The early diagnosis and treatment of fat embolism syndrome caused by the injuries in Wenchuan earthquake]. AB - OBJECTIVE: To analysis and discuss the early diagnosis and treatment of fat embolism syndrome caused by the injuries in Wenchuan earthquake happened in May 12th, 2008. METHODS: To carry out dynamic blood and oxygen analysis and blood routine examination for the 13 fat embolism syndrome patients wounded in the earthquake, and combine with clinical features and early diagnosis, inject hydrocortisone injection, dextran 40 glucose injection and Dan Shen Root injection. Meanwhile, pay attention to supporting the respiratory function and correcting hypoxemia. RESULTS: Of 13 patients, 12 cases were rescued successfully, and the successful rate was 92.3%, while one cases died from respirometric failure with fat embolism syndrome and secondary hemorrhagic pneumonia. CONCLUSIONS: The key to treating fat embolism syndrome is the early diagnosis and timely and accurate treatment. Supporting the respiratory function, correcting hypoxemia and early high-dose combination of hormone treatment are the effective treatment methods. PMID- 19134369 TI - [Analysis of the location, early diagnosis and treatment of osteofascial compartment syndrome resulted from Wenchuan earthquake]. AB - OBJECTIVE: To analyze the location, early diagnosis and treatment of osteofascial compartment syndrome (OCS). METHODS: There were 38 males and 29 females with age range of 8 - 69 years (mean 38.1 years). All 67 cases were suffered from Wenchuan earthquake happened in May 12th, 2008, of which 34 focuses with tibia-fibular fracture, 9 focuses with femoral fracture, 4 focuses with humeral fracture, and 13 focuses with radius and/or ulna fracture. The occurred place of OCS involves calf (41 focuses), legs (25 focuses), hands (2 focuses), feet (3 focuses), hip (19 focuses), forearms (15 focuses) and upper arms (10 focuses). The clinical symptoms and characteristics were closely observed and treated with decompression posterior to conservative treatment or with prompt open decompression. RESULTS: Of 67 cases (115 focuses), 82 focuses with secondary suture won satisfactory recovery of blood circulation and extremity function, 23 wounds were closed with secondary suture and skin grafting, 5 limbs were amputated due to avascular necrosis and 2 cases died (5 focuses OCS). Of 115 focuses 11 focuses obtain part function, 7 focuses lost join function. CONCLUSIONS: The proportion of some rare parts' pathogenesis of osteofascial compartment syndrome caused by the earthquake rises. Early diagnosis, close observation and open decompression are important for treatment of osteofascial compartment syndrome. PMID- 19134371 TI - [Research of laparoscopic radical cystectomy and urinary diversion]. PMID- 19134370 TI - [The crush syndrome patients combined with kidney failure after Wenchuan earthquake]. AB - OBJECTIVE: To retrospectively analysis the treatment characteristics of the systemic situation in patients with crush syndrome after Wenchuan earthquake happened in May 12th, 2008. METHODS: Forty-nine patients with crush syndrome and subsequent acute renal failure (ARF) due to the earthquake were treated in West China Hospital. All of patients had been rescued from buildings that collapsed in Wenchuan earthquake. The major associated injuries were in the low extremities and upper extremities. 49 patients developed ARF with increased concentrations of serum creatinine (mean 64 022 U/L) had underwent haemodialysis. Hyperkalaemia was seen in 9 patients and four of them underwent haemodialysis. 49 patients were administered hemodialysis. RESULTS: No patient died. All patients who suffered from the ARF were weaned from hemodialysis after admitted 7 to 35 days. Forty five extremities underwent amputations and 52 extremities had fasciotomy. CONCLUSIONS: Crush syndrome requires urgent recognition and prompt surgical treatment with simultaneous measures to control hyperkalemia and ARF. The authors believe that immediate intensive care therapy and multi-subjective coordination would have improved the survival rate. PMID- 19134372 TI - [The issues of laparoscopic radical prostatectomy]. PMID- 19134373 TI - [Compare of laparoscopic and open surgery for radical cystectomy with orthotopic ileal neobladder]. AB - OBJECTIVE: To compare the clinical therapeutic effect and complications of laparoscopic radical cystectomy with orthotopic ileal neobladder (LRC-INB) with open radical cystectomy with orthotopic ileal neobladder (ORC-INB). METHODS: A total of 171 patients were evaluated, including 63 cases with ORC-INB and 108 cases with LRC-INB from June 1994 to May 2007 at our institution. The parameters analyzed included perioperative data, postoperative complications, new bladder function and effect of tumor control. RESULTS: There was no significant difference in demographic characteristics of patients between these 2 groups. The mean operating time was 330 min in the LRC group and 310 min in the ORC group (P > 0.05). The mean blood loss was 320 ml in the LRC group and 1100 ml in ORC group (P < 0.001). The mean oral intake after operation was 2.4 days for LRC group and 4.5 days for ORC group (P < 0.001). No perioperative death was occurred in both groups. The complication rate was 18.5% in LRC group, while 30.0% in ORC group (P < 0.05). Twelve months after operation, the day-time and night-time continence rate were 90.7% and 82.6% for the LRC group, 88.3% and 81.6% for the ORC group respectively (P > 0.05). There was no significant difference of VOL, pressure and residual urine volume (RUV) of neobladder between these 2 groups. Surgical margin were tumor free for 107 cases except one T4 case in laparoscopic group had positive margin (P > 0.05). The mean number of removed lymph node were 12 and 8 in LRC and ORC group respectively (P < 0.05). The 2 years tumor free survival rate of the same stage or grade was no significant different (P > 0.05). CONCLUSIONS: LCR had advantages of less blood loss, shorter oral intake time, less postoperative complications, comparable continent rate and short-term tumor control with ORC. Long-term follow up is needed to confirm the oncological outcome. PMID- 19134374 TI - [Retroperitoneal laparoscopic resection for pheochromocytoma (experience of 107 cases)]. AB - OBJECTIVE: To evaluate the safety and efficacy of retroperitoneal laparoscopic resection for pheochromocytoma. METHODS: The clinical data of 107 cases of pheochromocytoma in PUMCH from 2003 to 2008 were analyzed retrospectively. There were 58 males and 49 females with an age range from 8 to 77 years (mean 44 years) in this cohort. One hundred and two cases were intra-adrenal and 5 extra-adrenal. Of the 102 intra-adrenal tumors, 43 tumors were located in left adrenal, 51 in right adrenal and 8 in both sides. All of the 5 extra-adrenal tumors were at para abdominal aorta. Retroperitoneal laparoscopic resection was performed for the 107 cases. This period was separated to 3 stages based on the degrees of the practical ability for retroperitoneal laparoscopic resection, such as tentative and exploratory stage, accumulative stage, and mature stage. RESULTS: At tentative and exploratory stage from June 2003 to December 2003, 10 cases underwent retroperitoneal laparoscopic surgery, of which 3 cases were converted to open surgery. The mean diameter was 4.2 cm (range in diameter from 2.5 cm to 6.0 cm). The mean operation time was 105 min (range from 60 min to 230 min). The mean volume of blood loss during operation was 620 ml (range from 150 ml to 1800 ml). At accumulative stage from January 2004 to December 2006, 66 cases underwent retroperitoneal laparoscopic surgery with none converted to open surgery. The mean diameter was 5.7 cm (range in diameter from 2.1 cm to 8.7 cm), and the diameter was above 6.0 cm in 19 cases. The mean operation time was 95 min (range from 40 min to 210 min). The mean volume of blood loss during operation was 350 ml (range from 50 ml to 1800 ml). At mature stage from January 2007 to June 2008, 31 cases, including 5 extra-adrenal pheochromocytomas, underwent retroperitoneal laparoscopic surgery. The mean diameter was 6.5 (range in diameter from 1.5 cm to 12.3 cm). The mean operation time was 75 min (range from 40 min to 160 min). The mean volume of blood loss during operation was 180 ml (range from 50 ml to 800 ml). No peri-operative death occurred. Follow up period was ranging from 1 to 62 months (the mean was 34 months), and 7 failed to be followed up, 3 cases recurred. And there was no distant metastases and death case. CONCLUSIONS: Retroperitoneal laparoscopic surgery for pheochromocytoma is feasible and safe. This procedure will be more and more performed as the advancement of the skill and accumulation of experience. The dimension, recurrence and location of tumor are not the absolute contraindication of retroperitoneal laparoscopic surgery for pheochromocytoma. PMID- 19134375 TI - [Transperitoneal laparoscopic adrenalectomy: surgical approach and outcome]. AB - OBJECTIVE: To investigate the efficiency and safety of transperitoneal laparoscopic adrenalectomy for the treatment of adrenal tumors and to describe surgical technique and management of intraoperative complications. METHODS: From February 2002 to April 2008, 24 male and 44 female consecutive patients with average age (53.4 +/- 12.1) years old were treated with transperitoneal laparoscopic adrenalectomy because of adrenal tumors. Of the patients, 27 cases had right adrenal tumors, 39 cases had left adrenal tumors and 2 cases had bilateral adrenal tumors. The average tumor size was (2.9 +/- 2.0) cm with the maximal diameter of 10 cm. We evaluated this technique in respect of operating time, estimated blood loss, complications during surgery, postoperative complications, duration of hospital stay and pathological results. RESULTS: All the surgeries had been completed successfully without conversion to open surgery. The average operating time was (157.7 +/- 51.5) min, the average estimated blood loss was (68.1 +/- 54.2) ml. No major complication happened during operation. The average drainage time was (2.6 +/- 1.5) days. The average post-operative hospital stay was (8.7 +/- 4.3) days. Four cases (6.0%) developed surgical field liquefaction and 2 cases (2.9%) experienced delayed closure of the drainage wound. CONCLUSIONS: Transperitoneal laparoscopic adrenalectomy is feasible and safe in the treatment of adrenal tumor with low risk of intra-operative and post operative complication. PMID- 19134376 TI - [Improve recovery of urinary continence following laparoscopic radical prostatectomy: a clinical report of 51 cases]. AB - OBJECTIVE: To present the clinical results of 51 patients of clinically localized prostate cancer treated by laparoscopic radical prostatectomy, especially recovery of urinary continence. To analysis how to improve recovery of urinary continence. METHODS: From February 2004 to March 2008, we performed LRP on 51 patients. All patients were diagnosed by pathological result preoperatively. T1a 1b 4 cases (8%), T1c 15 cases (29%), T2a 7 cases (14%), T2b 5 cases (10%), T2c 20 cases (39%). RESULTS: Forty-nine cases were treated by LRP successfully. There were two cases converted to open surgery. Urine leaking happened in 3 cases, but recovered without intervention. Urinary catheter duration was 14 - 45 days (mean 16 days). Six months and 12 months after LRP, incontinence happened 7/39 cases (17.9%) and 5/20 cases (25%) respectively. Complete incontinence happened in one case. The ratio of incontinence happened in the first 20 cases and the following 31 cases were 6/20 (30%) and 7/31 (22%) respectively. There were two cases had rectal injury. Both cases underwent colostomy. Duration of follow up were from 3 53 months (mean 17 months). Two cases recurred. One of them was treated with hormone therapy, the other one died of operation for lung metastasis. Other patients' serum total PSA were less than 0.2 microg/L. CONCLUSIONS: Extraperitoneal laparoscopic radical prostatectomy is efficient and safe approach for localized prostate cancer. The correct handling of pubic prostatic ligaments, external urethral sphincter and neurovascular bundle are important for recovery of urinary continence. And so does the experience of LRP. PMID- 19134380 TI - [A survey of bile duct injuries sustained during laparoscopic cholecystectomy]. AB - OBJECTIVE: To summarize the reasons for bile duct injury (BDI) after laparoscopic cholecystectomy (LC), and to determine the effect of multiple treatment after BDI. METHODS: A retrospective cohort study was performed. The medical records of 110 patients diagnosed with BDI after LC from October 1993 to November 2007, in ten large hospitals in Guangdong of China, were reviewed. RESULTS: Among 110 patients with BDI, 58 cases (52.7%) were local patients, whereas 52 cases (47.3%) were transferred from outside hospitals. Reasons for BDI following LC were: (1) Lack of experience of the LC operator (48.2%); (2) LC performed during acute cholecystitis (20.0%); (3) The structure of Calot triangle was unclear (15.5%); (4) Variable anatomical position (11.8%); (5) Intra-operation bleeding (4.5%). The commonest sites of injury were the choledochus and common hepatic duct (76.4%). Following BDI, endoscopic stenting or operative repair was performed in 106 patients. The overall success rate was 95.3% (101/106), with a mortality rate was 0.9% (1/106). Cholangitis occurred in 3.8% (4/106) cases. Choledocho enterostomy operation was performed in almost 60.0% (63/106) cases, and the success rate was 93.7% (59/63). Endoscopic stenting or operative repair was performed immediately following BDI in 23.6% (25/106) patients, the success rate was 100%; and within 30 days in 63.2% (67/106) patients. Eighty-eight out of 106 patients who underwent repair were successful following the first operative procedure. CONCLUSIONS: Factors such as an un-experienced operator and unclear anatomical position were causes of BDI following LC. Early operative repair should be regarded as the treatment of choice, in patients diagnosed with BDI. Early refer to an experienced hepatobiliary operator ensures a high success rate. PMID- 19134381 TI - [Retransplantation for hepatic artery complications after orthotopic liver transplantation]. AB - OBJECTIVE: To evaluate the efficacy and timing of re-transplantation for hepatic artery complications after orthotopic liver transplantation. METHODS: Between December 2003 and December 2006, the clinical data of 13 patients diagnosed as hepatic artery complications after liver transplantation were retrospectively analyzed. RESULTS: There were no significant difference in duration of operation and anhepatic phase between the initial transplantation and the second transplantation (P = 0.291, P = 0.312). However, intra-operative blood loss [(3.1 +/- 1.1) L vs. (1.5 +/- 0.9) L, P = 0.005] and intensive care unit stays [(4.3 +/ 1.8) d vs. (3.2 +/- 2.5) d, P = 0.015] were significantly increased in the re transplant patients. No perioperative mortality occurred. The postoperative mortality of liver re-transplantation was 38.5% (5/13) including acute renal failure in two patients, severe infection in two and heart infarction in one. The other 8 patients were followed from 6 months to 51 months, with a median survival time of 22.5 months. CONCLUSIONS: Liver re-transplantation is the only viable option for patients with irreversible graft dysfunction secondary to hepatic artery complications after liver transplantation. Proper indication and optimum time of re-transplantation, reasonable individual immunosuppression regime and effective perioperative care program contribute to the increase of the survival rate of the patients performed liver re-transplantation. PMID- 19134382 TI - [Predictive factors for level VI lymph node metastasis in papillary thyroid microcarcinoma]. AB - OBJECTIVE: To determine the predictive factors for level VI lymph node (LN) metastasis in thyroid papillary microcarcinoma (PTMC). METHODS: From November 2005 to January 2007, 86 patients with PTMC with a lateral cN0 were treated by thyroidectomy and elective level VI LN dissection without comprehensive lateral neck dissection. The data from the cases were analyzed retrospectively to determine the predictive factors for level VI LN metastasis. RESULTS: Forty cases (46.5%) of the patients were found with level VI LN metastasis. Tumor size (> or = 5 mm), thyroid capsular invasion or extracapsular invasion, enlarged level VI LN size (> or = 4 mm) were found significantly related to level VI LN metastasis on univariate analysis (P < 0.05). Tumor size (> or = 5 mm) and thyroid capsular invasion or extracapsular invasion were found to be independent predictive factors for level VI LN metastasis on multivariate analysis (P < 0.05). CONCLUSIONS: Thyroid capsular invasion or extracapsular invasion, tumor size (> or = 5 mm) were significantly associated with level VI LN metastasis in patients with PTMC. Elective neck dissection in level VI should be considered particularly in patients with thyroid capsular invasion or extracapsular invasion and a tumor greater than 5 mm. PMID- 19134383 TI - [Multivariate logistic regression analysis of postoperative severe complications and discriminant model establishment in gastric cancer post gastrectomy]. AB - OBJECTIVE: To investigate the main risk factors for postoperative severe complications, and establish Logistic regression model to predict severe complications in gastric cancer following gastrectomy. METHODS: The data of 1728 gastric cancer patients underwent gastrectomy between June 2001 and June 2007 were analyzed retrospectively. Logistic regression analysis was used to investigate the risk factors for postoperative severe complications in those patients. RESULTS: Postoperative severe complications were associated with extent of lymph node dissection (D(2)(+)-D(3)), chronic obstructive pulmonary disease (COPD), invasion to the adjacent organ, combined organ resection, extent of lymph node dissection (D(2)), diabetes mellitus (DM), TNM staging IV, heart diseases, malnutrition, surgeon's operative volume, operative time, blood loss and age. The Logistic regression model was P = 1/[1+e((14.806-2.523X1-1.792X2-1.558X3-1.551X4 1.270X5-1.150X6-1.101X7-0.981X8-0.817X9-0.657X10-0.578X11-0.542X12-0.309X13))]. A testing sample showed that the accuracy, sensitivity and specificity of the Logistic model were 72.5%, 70.0% and 75.0%, respectively. CONCLUSIONS: The extent of nodal dissection (D(2)(+)-D(3)), COPD, invasion to the adjacent organ, combined organ resection, extent of nodal dissection (D(2)), diabetes mellitus, TNM staging IV, heart diseases, malnutrition, surgeon's operative volume, operative time, blood loss and age are the independent risk factors associated with severe complications in gastric cancer post gastrectomy. The Logistic regression model based on these factors is reliable in predicting the severe complications. PMID- 19134384 TI - [Assessment of energy expenditure and body composition in cancer patients]. AB - OBJECTIVE: To demonstrate the changes of resting energy expenditure (REE), substrate metabolism and body composition in cancer patients. METHODS: From September 2004 to March 2008, REE, carbohydrate oxidation (CO) and fat oxidation (FO) in 936 cancer patients and 840 control subjects were measured by indirect calorimetry. Bioelectrical impedance appliance was applied to assess intracellular fluid, extracellular fluid, fat mass (FM) and fat free mass (FFM) in the two groups. RESULTS: No difference in REE was found between the cancer patients and non-cancer patients [(1452.2 +/- 196.4) kcal/d vs. (1429.5 +/- 182.6) kcal/d, P = 0.136]. But REE/FFM and REE/pREE were elevated in cancer patients than in controls (all P < 0.05). Of the cancer patients, 48.6% were hypermetabolic, 42.9% normal and 8.5% hypometabolic, while those were 22.5%, 58.5% and 19.0% in controls. Cancer patients had higher FO [(77.8 +/- 11.3) g/min vs. (67.1 +/- 12.1) g/min, P = 0.000], lower CO and npRQ [(68.7 +/- 10.5) g/min vs. (88.8 +/- 12.1) g/min, P = 0.000; 0.782 +/- 0.012 vs. 0.810 +/- 0.014, P = 0.000]. Cancer patients exhibited lower FM and FFM [(14.9 +/- 4.5) kg vs. (18.4 +/- 5.2) kg, P = 0.000; (44.4 +/- 7.2) kg vs. (46.1 +/- 8.1) kg, P = 0.008]. CONCLUSIONS: Elevated REE is common in cancer patients. Substrate metabolism of the cancer patients features in increased FO, decreased CO and npRQ, which is correlated with the elevated REE. FM and FFM loses in proportion in cancer patients. PMID- 19134385 TI - [Tricuspid valve replacement and anticoagulation therapy: a report of 70 cases]. AB - OBJECTIVE: To evaluate the operative indication, the choice of valve prostheses, and the method of anticoagulation therapy of tricuspid valve replacement (TVR). METHODS: From May 1998 to January 2008, 70 patients underwent TVR. There were 59 cases of rheumatic heart diseases. The operations included mitral and tricuspid valve replacement for 37 cases, triple valve replacement for 18 cases, isolated TVR for 13 cases, and tricuspid and aortic valve replacement for 2 cases. All the patients received oral anticoagulant therapy (warfarin) 2 d after the operations. RESULTS: There were 3 in-hospital deaths (4.3%) and 1 late death (0.4%). The follow-up rate was 88.1%, and the cumulative follow-up was 243.5 patient-years (pty). The anticoagulation-related event rate was 2.9% pty, and the mean INR value of 643 out-patient samples was 1.87 +/- 0.68. Post-operative heart function NYHA classification: 52 cases in class I to II, and 7 cases in class III. CONCLUSIONS: TVR should be indicated for severely damaged and deformed tricuspid valve of rheumatic heart diseases. Bileaflet mechanical valve is a suitable prosthesis for TVR. The optimal anticoagulation therapy intensity of TVR needs to be investigated. PMID- 19134386 TI - [Long-term results of surgical repair of ruptured sinus of aortic sinus aneurysm]. AB - OBJECTIVE: To summarize the clinic characteristics and effect of surgical repair of ruptured aortic sinus aneurysm. METHODS: From September 1997 to September 2007, 43 patients with ruptured aortic sinus aneurysm underwent surgical procedures. There were 32 male and 11 female patients. The age ranged from 11 to 50 years old with a mean of (29.0 +/- 11.5) years old. The origins of rupture were the right coronary sinus in 34 patients and the noncoronary sinus in 9 patients. The aneurysms ruptured into the right ventricle in 30 patients, the right atrium in 8 patients, the right ventricle and right atrium in 3 patients, and the ventricular septum and then the right ventricle in 2 patients. Associated cardiac anomalies included ventricular septal defect in 26 patients, aortic regurgitation in 15 patients, infectious endocarditis in 8 patients, tricuspid regurgitation in 6 patients, atrial septum defect in 4 patients, mitral valve regurgitation in 2 patients, patent ductus arteriosus in 2 patients, and pulmonary valve vegetation in 1 patient. All the patients underwent the repair of ruptured aortic sinus aneurysm and correction of associated anomalies with cardiopulmonary bypass. RESULTS: There were no deaths after the operation and during the follow-up. The complications, including acute heart failure and III atrioventricular block, occurred in 5 patients. Follow-up was 6 to 120 months with a mean of (68.0 +/- 17.7) months. Two patients underwent reoperation for aortic valve replacement at the 6(th) and 8(th) year after the first operation. There were 2 patients which the aortic regurgitation deteriorated from grade I to II. CONCLUSIONS: Repair of ruptured aortic sinus aneurysm presents a satisfactory result. Aggressive treatment in early time, prevention of post-operative complications and long-term follow-up are recommended in the treatment for patients of ruptured aortic sinus aneurysm with infectious endocarditis and aortic regurgitation. PMID- 19134387 TI - [Intraoperative esophageal manometry employed in the course of Heller's cardia myotomies]. AB - OBJECTIVE: To study the changes of lower esophageal sphincter (LES) high-pressure zone, and to determine the accurate length of myotomy on the esophageal and gastric sides. METHODS: There were 15 patients undergoing the Heller's cardia myotomies and Toupet fundoplications from May 2006 to December 2007. Among them, 9 patients were female and 6 was male. The age ranged from 28 to 61 years old, and the disease duration ranged from 6 months to 9 years. The intraoperative oesophageal manometry underwent in the surgical procedures to investigate the changes of the lower esophageal sphincter pressure and the length of myotomy. RESULTS: There was no postoperative death. After (5.3 +/- 1.5) cm of esophageal side myotomy and (0.8 +/- 0.4) cm of gastric side myotomy, the mean LES pressure decreased from (33.6 +/- 13.3) mm Hg (1 mm Hg = 0.133 kPa) to (9.7 +/- 4.6) mm Hg and (4.8 +/- 3.1) mm Hg respectively (P < 0.05). The lower esophageal sphincter length ranged from 5 to 8 cm. CONCLUSION: Intraoperative esophageal manometry helps determine the accurate myotomy length of myotomy on the esophageal and gastric sides of the gastroesophageal junction and provides valuable information for the Heller's myotomy. PMID- 19134388 TI - [Construction of pEGFP-HBx gene recombinant plasmid and establishment of stable transfected HBx gene liver oval cell strain]. AB - OBJECTIVE: To construct pEGFP-HBx eukaryotic expression plasmid and establish stable and effective transfected rat oval cell (LE/6) strain expressing EGFP-HBx fusion protein to explore the roles of HBx gene and oval cell in carcinogenesis of hepatocellular carcinoma (HCC). METHODS: HBx gene with EcoRI and Hind III endo enzyme sites was obtained by using PCR from plasmid pcDNA3.1-HBx. The purified HBx gene fragment was inserted into pEGFP-N1 expression vector, and the recombinant plasmid pEGFP-HBx was identified by restriction endonuclease and DNA sequencing analysis. LE/6 cells were transfected with recombinant pEGFP-HBx by lipofectamine reagent. Resistant to G418 clones were selected, and expression of EGFP-HBx fusion protein in clones were examined directly with fluorescence microscope, and these clones were isolated and proliferated. The expression of HBx was detected by RT-PCR analysis and immunocytochemistry. RESULTS: Plasmid pEGFP-HBx has whole HBx gene base and correct reading frame as indicated by restriction endonuclease and DNA sequencing analysis. After transfecting with pEGFP-HBx plasmid, LE/6 cell clones expressing EGFP-HBx fusion protein were obtained. RT-PCR analysis and immunocytochemistry showed that HBx gene was only expression in transfected pEGFP-HBx cells. CONCLUSIONS: The pEGFP-HBx recombinant expression vector was successfully constructed, and the stable transfected LE/6 strain expressing EGFP-HBx fusion protein was successfully established. It will be helpful in the further study on the roles of HBx and liver oval cell in carcinogenesis of HCC. PMID- 19134400 TI - [Improving diagnosis and treatment of lung cancer with state-of-the-art medicine]. PMID- 19134401 TI - [On the current situation and the diagnosis of early lung cancer]. PMID- 19134389 TI - [In vitro comparison of antibacterial properties of plasma sprayed wollastonite coatings loading silver and gentamicin]. AB - OBJECTIVE: To develop antibacterial coatings for orthopedic implants with a sustained release of drugs. METHODS: Wollastonite coatings were deposited on the titanium substrates by an atmospheric plasma spray system. After soaking in weight percent of 5% AgNO(3) solution for 24 h, the wollastonite coatings loading silver were obtained. Gentamicin were loaded on the wollastonite coatings by collagen grafting process. The release rates of drugs from wollastonite coatings were investigated by the in vitro solution soaking test. One strain of S. aureus was used in zone of inhibition test to evaluate the antibacterial properties of drug loaded wollastonite coatings, and the cell culture test was used to evaluate their cytotoxicity. RESULTS: Silver and gentamicin loaded wollastonite coatings were successfully prepared. The release of silver ions from the silver loaded wollastonite coatings lasted 50 d in deionized water, effectively inhibiting the growth of S. aureus for 40 d. While an initial burst release of gentamicin was found during the in vitro solution soaking test. The gentamicin released from gentamicin loaded wollastonite coatings can inhibit the growth of S. aureus for 18 d. Both the two kinds of antibacterial wollastonite coatings showed no adverse effect on cellular adhesion, proliferation and alkaline phosphatase expression. CONCLUSIONS: Compared with gentamicin loaded wollastonite coatings, silver loaded wollastonite coatings may have more promising clinical applications due to the even and long-time antibacterial agent release. PMID- 19134402 TI - [Countermeasures for resistance to chemotherapeutic agents in lung cancer]. PMID- 19134403 TI - [Genomic approaches to tailored therapy in non-small cell lung cancer]. PMID- 19134404 TI - [The detection by denaturing high performance liquid chromatography of epidermal growth factor receptor mutation in tissue and peripheral blood from patients with advanced non-small cell lung cancer]. AB - OBJECTIVE: To study the application of denaturing high performance liquid chromatography (DHPLC) as a screening tool in detecting plasma and matched tissue epidermal growth factor receptor (EGFR) mutations for advanced non-small-cell lung cancer (NSCLC). METHODS: Plasma DNA samples and matched tumors from 230 cases of NSCLC were analyzed for EGFR mutations in exons 19 and 21 using DHPLC. The mutations in the plasma samples and the matched tumors were compared, and the association between EGFR mutations and the clinicopathological features were evaluated. RESULTS: Mutation of EGFR was found by DHPLC to be 33.5% (77/230) in tissues and 34.3% (79/230) in matched peripheral blood samples. Consistency of EGFR mutation status between tissues and matched plasma DNA was confirmed (kappa is 0.74, P < 0.01). The sensitivity and specificity of DHPLC for detecting EGFR mutation were 96.9% and 91.9%, respectively (kappa is 0.88). EGFR mutations in both tissue and blood was correlated with histology type (OR = 3.38, 95% CI 1.81 6.36, P < 0.05) and smoking status (OR = 1.61, 95% CI 1.13 - 2.28, P < 0.05), but no association with age, sex and stage was found (P > 0.05). CONCLUSION: The detection of EGFR mutation is highly consistent in tissues and in plasma DNA samples. DHPLC may serve as a preliminary screening tool for detecting EGFR mutations. PMID- 19134405 TI - [Evaluation of the diagnostic role of endobronchial ultrasonography for peripheral lung cancer]. AB - OBJECTIVE: To describe the endobronchial ultrasonographic characteristics and the cut-off value for diagnosis of peripheral lung cancer, and therefore to evaluate its diagnostic value. METHODS: During June 1st, 2005 and June 30th, 2006, 78 patients with peripheral pulmonary lesions were enrolled. The lesions were all detectable by endobronchial ultrasonography (EBUS) and a final diagnosis was made. The endobronchial ultrasonographic structure of peripheral pulmonary lesions were analyzed, differentiated and classified into malignant or benign groups. RESULTS: According to the result of binary multivariable logistic regression analysis on the 9 variables and by calculating the area under ROC curve, 5 variables were found to be useful in predicting the presence of malignancy: (1) clear borderline; (2) internal hypoechoic echo; (3) heterogeneous pattern; (4) without internal hyperechoic dots and linear arcs; (5) adjacent blood vessels representing shift, narrow or break-off. The equation of malignancy probability for any patient was: P = 1/[1 + e(-) (6.321-3.097X(2)-1.537X(1) + 1.898X(5) + 2.390X(3) + 3.003X(4))], X(1) for borderline; X(2) for internal hyperechoic dots and linear arcs; X(3) for adjacent blood vessels; X(4) for internal echo intensity; X(5) for internal echo distribution. The areas of ROC curve illustrated that multivariable logistic regression model discriminated benign and malignant lesions better than univariable logistic regression. The optimal cut-off value of the malignancy probability, which was greater or equal to 0.52 according to the ROC curve. This model gave a sensitivity and specificity of 87.2% and 80.6%, and the accuracy was 85.9%. CONCLUSIONS: Endobronchial ultrasonographic characteristics of peripheral lung cancer included clear borderline, internal hypoechoic echo, heterogeneous pattern, without hyperechoic dots and linear arcs, and adjacent blood vessel shift, narrow or break-off. Multivariable logistic regression model discriminated benign and malignant lesions better than univariable logistic regression. Combination of multiple variables increases the sensitivity, specificity and accuracy for prediction of malignancy. PMID- 19134406 TI - [Anti-tumor immune response of dendritic cells transfected with wild-type p53 gene]. AB - OBJECTIVE: To explore the anti-tumor immune responses of dendritic cells (DCs) loaded with intact wild-type p53 to mice challenged with tumor cells expressing p53 genes with mutations at different sites. METHODS: Ad-p53-DC immunization function was assessed by the expression of surface molecules and allogeneic MLR. DCs derived from bone marrow were transduced with adenovirus or a human wild-type p53 containing recombinant adenovirus (Ad-DC and Ad-p53-DC) and immunized C57BL/6 mice. Splenocytes were separated and cell cytotoxicity was measured against tumor cells expressing mutant p53 (MethA, D459 and P815) in a standard 6-h(51)Cr release assay. Effector and target cells were incubated in the presence of anti CD(4) or anti-CD(8) antibody. Ad-p53-DC was immunized in control Ad-DC before or after mice were challenged with either D459 tumor or with MethA sarcoma cells to observe whether immune response would provide tumor protection. RESULTS: Immunization with Ad-p53-DC developed significantly higher substantial CTL responses against Ad-p53-P815, D459 and MethA cells (effectors: target cells = 50:1), (27.8 +/- 3.4)%, (23.5 +/- 2.7)%, (58.3 +/- 9.2)% than with Ad-DC (9.3 +/- 1.8)%, (4.6 +/- 1.0)%, (23.5 +/- 3.7)% (t(d) = 5.79, 3.68, 5.02, all P < 0.05). In Ad-p53-DC immunized mice, anti-CD(8) antibody blocked the cytotoxicity against Ad-p53-P815 (26.7 +/- 2.8)% or D459 (6.1 +/- 1.2)%, but not anti-CD(4) antibody [(59.8 +/- 4.6)%, (18.9 +/- 2.4)%, t(d) = 8.79 or 9.18, all P < 0.05]. Ad-p53-DC immunization provided complete tumor protection in 80% of mice challenged with D459 and in 70% of mice challenged with MethA, while none protected in Ad-DC immunization group (chi(2) = 6.72, 5.86, all P < 0.05). Treated with Ad-p53-DC after D459 inoculation subcutaneously, mice were killed due to the bulky tumor more than 2 weeks later than the mice in the Ad-DC treatment group during 7 week observation (chi(2) = 9.48, P < 0.05). CONCLUSION: DCs transfected with 100 MOI Ad-p53 induced intense CTL responses against P815, D459 and MethA. This CTL response is mediated by CD(8)(+) T cells. Treatment with Ad-p53-DC significantly developed tumor immunology and slowed the growth of established tumors. PMID- 19134408 TI - [Impulse oscillometry for estimation of airway obstruction]. AB - OBJECTIVE: To explore the application of impulse oscillometry (IOS) in the estimation of airway obstruction and to evaluate the correlation between spirometry indices and IOS parameters. METHODS: From Nov.2007 to May 2008, spirometry and IOS measurements were performed in 100 participants (male 72, female 28). FEV(1), FVC, FEV(1)/FVC, airway resistance at 5 Hz (R(5)), airway resistance at 20 Hz (R(20)), central resistance (Rc) and peripheral resistance (Rp) of structural parameters interpretation graph, FEV(1)% pred, R(5)% pred, R(20)% pred, and FEV(1)/FVC were analyzed. Correlations between spirometry and IOS parameters were studied. RESULTS: All participants had satisfactory impulse oscillometry results. R(5) and Rp were significantly increased in FEV(1)/FVC < 70% group [(5.3 +/- 2.1) and (6.2 +/- 2.9) cm H(2)OxL(-1)xs(-1) (1 cm H2O = 0.098 kPa)], respectively. There was significant negative correlation between FEV(1) and R(5) and Rp (correlation coefficient was -0.38 and -0.47 respectively, all P < 0.01). There was also negative correlation between FVC and R(5) and Rp (correlation coefficient was -0.28 and -0.37, respectively, all P < 0.01). A significant negative correlation between FEV(1)% pred, FVC% pred, FEV(1)/FVC% pred and R(5)% pred was also observed (correlation coefficient -0.49, -0.39 and 0.43, respectively). CONCLUSIONS: IOS parameters can be used to evaluate airway obstruction. Among IOS parameters, R(5) was the most sensitive, which was also significantly correlated with spirometric parameters. PMID- 19134407 TI - [Detection of SLC34A2 in patients with pulmonary alveolar microlithiasis and the effect of SLC34A2 on transportation of calcium and phosphate in human alveolar epithelial cells]. AB - OBJECTIVE: To detect the mutation of SLC34A2 in patients with pulmonary alveolar microlithiasis and to study the effect of SLC34A2 on transportation of calcium and phosphate in human alveolar epithelial cell (A549) cells. METHODS: The gene SLC34A2 was detected by segmentation-PCR and gene sequencing. RNA was obtained by Trizol from fresh lung tissues and the target gene was acquired by RT-PCR. Eukaryotic expression of recombinant pcDNA3.1(+)-SLC34A2 was constructed and SLC34A2 was transfected to A549 cells by liposome. The expression of SLC34A2 mRNA was detected by RT-PCR, and the content of calcium and phosphate of the extracellular fluid was measured by commercial kits. The cell experiments consisted of 3 groups including a control group (5 x 10(5)/well, one well), a blank group (5 x 10(5)/well, one well), a transfection group (5 x 10(5)/well, four wells). Every experiment was repeated 6 times. RESULTS: No mutation was found in patients with pulmonary alveolar microlithiasis. SLC34A2 cDNA was successfully amplified and the eukaryotic expression recombinant pcDNA3.1(+) SLC34A2 was successfully constructed. The amount of SLC34A2 mRNA of the transfected cells was significantly higher (2.48 +/- 0.45), compared to the control cells (0.55 +/- 0.07) and the blank cells (0.60 +/- 0.06), q = 16.25, 15.78, all P < 0.01. The content of calcium and phosphate in the supernatant of the transfected cells was lower [(0.110 +/- 0.016) mmol/L, (3.8 +/- 0.4) mmol/L], compared with the control [(0.254 +/- 0.047) mmol/L, (7.3 +/- 0.8) mmol/L] and the blank (0.262 +/- 0.041) mmol/L, (7.1 +/- 0.4) mmol/L], q = 8.657 - 13.892, all P < 0.01. CONCLUSIONS: In human lung alveolar epithelial cells, the content of calcium and phosphate in cell supernatant decreased with increased amount of SLC34A2 mRNA. Mutation of SLC34A2 may not be at the DNA level. PMID- 19134409 TI - [PKCalpha-ERK1/2 cascade is involved in up-regulation of cyclinD1 and P21(cip1) in human airway smooth muscle cells sensitized by sera from atopic asthmatics]. AB - OBJECTIVE: To explore the role of PKCalpha-ERK1/2 cascade in PMA induced up regulation of cyclinD1 and P21(cip1) in human airway smooth muscle cells (HASMCs) sensitized by sera from atopic asthmatics. METHODS: HASMCs in cultures were passively sensitized by 10% serum from asthmatic patients and were randomly divided into five groups: the control group, PMA treated group, PMA and PKCalpha mismatched Oligodeoxynucleotides (PKCalpha-mmODN) treated group, PMA and PKCalpha antisense Oligodeoxynucleotides (PKCalpha-asODN) treated group, PMA and U0126 (MAP Kinase Kinase inhibitor)treated group. The expression of p-PKCalpha, ERK1/2, p-ERK1/2, cyclinD1 and P21(cip1) protein were determined by western blotting. The proliferation of HASMC was examined by cell cycle analysis and MTT colorimetric assay. RESULTS: Compared with the control group, the expression of p-PKCalpha and ERK1/2, p-ERK1/2 protein increased, the expression of cyclinD1, P21(cip1) protein increased correspondingly (the A value % control was 2.10 +/- 0.29, 1.67 +/- 0.19, 2.20 +/- 0.27, 1.99 +/- 0.22 and 3.11 +/- 0.29 respectively; q value was 9.87, 7.06, 10.57, 11.10 and 20.33 respectively; all P < 0.05) in PMA treated group, and cells proliferation [the percentage of cells in S phase was (30.3 +/- 2.4)%, A(490) value was 0.80 +/- 0.06] enhanced significantly compared with those [the percentage of cells in S phase was (13.9 +/- 2.6)%, A(490) value was 0.41 +/ 0.04] of the control group (q = 6.07, 12.63; all P < 0.05). In PMA and PKCalpha asODN treated group, the level of p-PKCalpha decreased, the expression of ERK1/2, p-ERK1/2 and the expression of cyclinD1, P21(cip1) decreased correspondingly (the A value % control was 1.23 +/- 0.19, 1.34 +/- 0.18, 1.52 +/- 0.20, 1.45 +/- 0.18 and 1.49 +/- 0.18 respectively; q value was 7.49, 3.58, 5.97, 6.06 and 15.65 respectively; all P < 0.05), and cells proliferation reduced significantly [the percentage of cells in S phase was (21.2 +/- 2.8)%, A(490) value was 0.51 +/- 0.04; q = 6.07, 12.63; all P < 0.05], as compared with those of the PMA treated group. In PMA and U0126 treated group, the level of p-PKCalpha had no significant change (A value was1.99 +/- 0.18, q = 0.94, P > 0.05), but the levels of ERK1/2, p-ERK1/2 decreased, the expression of cyclinD1, P21(cip1) reduced (the A value % control was 0.95 +/- 0.21, 1.15 +/- 0.19, 1.37 +/- 0.15 and 1.96 +/- 0.21 respectively; q value was 7.79, 9.16, 6.92 and 11.16 respectively; all P < 0.05), and cells proliferation reduced significantly [the percentage of cells in S phase was (22.0 +/- 3.2)%, A(490) value was 0.49 +/- 0.03; q = 5.51, 13.45; all P < 0.05], as compared with those of the PMA treated group. CONCLUSION: ERK1/2 is one of the downstream regulators of PKCalpha, and PKCalpha-ERK1/2 cascade is involved in PMA induced up-regulation of cyclinD1 and P21(cip1) and proliferation in HASMC sensitized by sera from atopic asthmatics. PMID- 19134411 TI - [Progress of proteomics in diagnosis and treatments of lung cancer]. PMID- 19134410 TI - [Effect of NADPH oxidase activity inhibitor apocynin on blood pressure in rats exposed to chronic intermittent hypoxia and the possible mechanisms]. AB - OBJECTIVE: To investigate the effect of NADPH oxidase activity inhibitor apocynin on the blood pressure of rats exposed to chronic intermittent hypoxia (CIH). METHODS: Thirty healthy male SD rats were randomly divided into 3 groups of 10 each, a CIH group, an apocynin treatment group and a control group. The systolic blood pressure (SBP) was measured with tail-cuff method. RT-PCR and Western blot were used to examine mRNA and protein expression of NADPH oxidase subunit p22phox. The level of MDA, SOD, NO and .O(2)(-) were detected by colorimetric method. RESULTS: Compared to the normal group, the expression of p22phox mRNA and protein were significantly increased in CIH group (q = 3.202, 6.522, P < 0.05). There were no statistic difference of p22phox mRNA and protein expression between CIH group and apocynin treatment group (P > 0.05). Compared with those of CIH group, the levels of MDA, .O(2)(-) and the systolic blood pressure decreased significantly (P < 0.05), while SOD and NO activity increased significantly (q = 6.960, 4.385, P < 0.05) in apocynin treatment group. CONCLUSIONS: These results indicate that NADPH oxidase up-expression is closely associated with hypertension in OSAHS. Inhibition of NADPH oxidase activity may be hopefully served as a useful strategy for prevention and treatment of OSAHS-induced hypertension. PMID- 19134412 TI - [Inhaled combination therapy with corticosteroids and long-acting beta-2 agonists in chronic obstructive pulmonary disease]. PMID- 19134413 TI - [Mycobacterium thermoresistibile: notable non-tuberculous mycobacteria]. PMID- 19134428 TI - Sensitivity to para-phenylenediamine and intolerance to hydrochlorothiazide. AB - para-Phenylenediamine (PPD) is a contact allergen that cross-reacts with a variety of medications, including thiazide diuretics and sulfonamides. We present the case of a 52-year-old African American atopic woman who came for evaluation of a severe eyelid dermatitis and recurrent outbreaks of acneiform (follicular based) pruritic papules on her face, chest, and back. These symptoms presented after the patient was started on hydrochlorothiazide for hypertension. The patient also reported a history of eyelid dermatitis associated with "black hair dye." Patch testing revealed a positive reaction to para-phenylenediamine (1+) and Disperse Blue 106 (1+). It was suspected that the patient might be demonstrating cross-reactivity to hydrochlorothiazide and a flare-up reaction of the eyelid dermatitis. After her cardiologist changed her blood pressure medication, the patient's eyelid dermatitis and eruption cleared. This case highlights the impact of PPD sensitization on the medical care of a patient with hypertension and the importance of choosing safer PPD-free alternatives and minimizing PPD exposures. PMID- 19134429 TI - Patch-test-induced "flare-up" reactions to neomycin at prior biopsy sites. AB - "Recall dermatitis" classically refers to the uncommon phenomenon of chemotherapy induced reactivation of skin damage originally induced by radiotherapy months or years earlier. We compare this condition to ectopic flare-up reactions presenting as dermatitis at existing or previous sites by an inducing agent. Enhanced sensitization of an existent allergen by patch testing is rarely described. We describe a case of a 61-year-old man's developing localized allergic contact dermatitis at previous neomycin-treated sites following the diagnosis of neomycin sensitivity on patch testing, which we attribute to a "flare-up" phenomenon. PMID- 19134430 TI - Cocamidopropyl betaine. PMID- 19134431 TI - Glyceryl monothioglycolate. PMID- 19134432 TI - Nickel sensitization, hand eczema, and loss-of-function mutations in the filaggrin gene. AB - Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease. Recently, it was demonstrated that a large proportion of individuals with AD have an epidermal expression deficiency of filaggrin (FLG). This unique finding may have great implications for our understanding of nickel sensitization because nickel is chelated in the epidermis and perhaps to FLG. This review aims to briefly present the current knowledge about nickel sensitization in relation to FLG deficiency and speculate on its possible implications. The new knowledge concerning loss-of-function mutations in the FLG gene (the lack of specific nickel-chelating power in the stratum corneum and a generally defective skin barrier) suggests that an additive effect from irritants and nickel may aggravate hand eczema in individuals with loss-of-function mutations in the FLG gene. This hypothesis calls for a reevaluation of the potential risk of nickel sensitization through the establishment of cohorts with and without such mutations. PMID- 19134433 TI - Novel antibacterial and emollient effects of coconut and virgin olive oils in adult atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) skin is dry and readily colonized by Staphylococcus aureus (SA). Coconut and olive oils are traditionally used to moisturize and treat skin infections. OBJECTIVE: To compare virgin coconut oil (VCO) and virgin olive oil (VOO) in moisturizing dryness and removing SA from colonized AD skin. METHODS: This was a double-blind controlled trial in two outpatient dermatology clinics with adult AD patients who were diagnosed by history, pattern, evolution, and skin lesions and who were randomized to apply VCO or VOO twice daily at two noninfected sites. SA cultures, photography, and objective-SCORAD severity index (O-SSI) scoring were done at baseline and after 4 weeks. RESULTS: Twenty-six subjects each received VCO or VOO. Of those on VCO, 20 were positive for SA colonies at baseline versus 12 on VOO. Post intervention, only 1 (5%) VCO subject remained positive versus 6 (50%) of those on VOO. Relative risk for VCO was 0.10, significantly superior to that for VOO (10:1, p = .0028; 95% CI, 0.01-0.73); thus, the number needed to treat was 2.2. For the O SSI, the difference was not significant at baseline (p = .15) but was significantly different post treatment (p = .004); this was reduced for both oils (p < .005) but was greater with VCO. CONCLUSION: VCO and monolaurin's O-SSI reduction and in vitro broad-spectrum activity against SA (given clinical validity here), fungi, and viruses may be useful in the proactive treatment of AD colonization. PMID- 19134434 TI - The effect of a transition to powder-free latex gloves on workers' compensation claims for latex-related illness. AB - BACKGROUND: Adverse reactions to natural rubber latex gloves are an important cause of workers' compensation claims among health care personnel. In an effort to reduce latex-related illness, our institution transitioned to powder-free latex gloves in 2001. OBJECTIVE: To examine the effect of this transition on the number of workers' compensation claims for latex-related illness. METHODS: A review of claims data from 1997 to 2005 was conducted. RESULTS: The incidence of claims for latex-related illness was significantly lower following the switch to powder-free gloves. CONCLUSIONS: The switch to powder-free latex gloves was associated with a significant decrease in workers' compensation claims for latex related illness. The cost of gloves increased but was partially offset by a decrease in workers' compensation payments and operating room expenses. PMID- 19134435 TI - Frequency and main sites of allergic contact dermatitis caused by nail varnish. AB - BACKGROUND: Liquid nail varnish has been used since 1919, and allergic contact dermatitis (ACD) has been recognized for at least 80 years, but it is difficult for nonspecialists to identify this condition. OBJECTIVES: (1) To verify the frequency of ACD from nail varnish in patients with a presumptive diagnosis of contact dermatitis seen at an outpatient clinic, (2) to characterize the groups studied according to site of skin disorder, and (3) to determine the main sensitizer related to varnish. METHODS: Patients with a final diagnosis of ACD caused by nail varnish were assessed by means of retrospective analysis of medical charts and protocols used in the clinic from January 1996 to December 2006. Patch tests with the Brazilian standard series and a complementary series were applied to all patients. RESULTS: Diagnosis of ACD from nail varnish was made in 8% of cases (157 of 1,971). The most affected sites were the face and neck; however, involvement of some uncommon areas, such as periungual and perianal regions, was also observed. CONCLUSIONS: ACD from nail polishes is a common event and recognition of the condition must be improved. Toluenesulfonamide formaldehyde resin (TSFR) was the most common allergen in the group studied. PMID- 19134436 TI - Sorbitan sesquioleate, a common emulsifier in topical corticosteroids, is an important contact allergen. AB - BACKGROUND: In the past, sorbitan sesquioleate (SSO) was reported as an uncommon allergen, but recent data suggest SSO may be an important sensitizer. OBJECTIVE: To present data on 13 of 112 dermatitis patients who reacted to SSO and/or sorbitan monooleate (SMO) on patch testing. METHODS: A retrospective data analysis was conducted on data from 112 dermatitis patients patch-tested from December 2006 to May 2007. All patients were tested with a modified North American Contact Dermatitis Group standard series, a cosmetic series, and a fragrance series. RESULTS: Of 112 patients, 10 (8.9%) reacted to SSO, 1 (0.9%) to SMO, and 2 (1.8%) to both. Nine of 12 (75%) SSO-positive patients were using topical corticosteroids emulsified with sorbitan derivatives or sorbitol; 2 of the 13 sorbitan-allergic patients were allergic to one or more corticosteroid screening chemicals tested. CONCLUSION: SSO is a common emulsifier derived from sorbitol and is used in many high- to super-potent corticosteroids. It has only recently been identified as an important contact allergen. The high prevalence of reactions to sorbitol derivatives in this small group of patients suggests that these chemicals may be sensitizing when applied to dermatitic skin. Larger studies should be conducted to confirm these findings. PMID- 19134437 TI - Eyelid dermatitis: contact allergy to 3-(dimethylamino)propylamine. AB - We present the case of a 42-year-old woman with intractable eyelid dermatitis. Patch testing revealed sensitization to 3-(dimethylamino)propylamine (DMAPA). DMAPA is an important etiology of allergic contact dermatitis of the eyelids and face but is easily missed even with expanded-series patch testing. We also review the most common causative allergens in eyelid dermatitis cited in the literature over the past decade. DMAPA is a reagent used in the formation of cocamidopropyl betaine (CAPB), a common additive to liquid soaps, shampoos, and other cleansing products because of its utility as a surfactant. Beginning in the 1980s, reports of allergy to CAPB surfaced in the literature. Ultimately, a majority of patch testing studies have shown that clinical allergy to CAPB-containing products actually reflects allergy to contaminant DMAPA in most cases. Amidoamine, another intermediate in the formation of CAPB, may also be implicated through a proposed mechanism of conversion to DMAPA in the skin. When patch-testing for eyelid and facial dermatitis, it is crucial to test with DMAPA directly, not just with CAPB; unlike commercial-grade CAPB, the CAPB in patch test kits is ultrapure and does not contain contaminant DMAPA. PMID- 19134438 TI - Contact dermatitis in the bowling pro shop. AB - A 29-year-old bowling pro shop operator presented with recurrent disseminated erythematous and edematous patches and plaques. Patch-testing with the North American Contact Dermatitis Group (NACDG) standard series yielded ++ and + reactions to epoxy resin at 2 and 7 days, respectively. We could find no other report of epoxy resin contact allergy in a bowling pro shop operator. PMID- 19134439 TI - Sorbitan sesquioleate: an emerging contact allergen. AB - Sorbitol-based emulsifiers such as sorbitan sesquioleate (SSO) are commonly used in topical corticosteroids, topical antibiotics, topical antifungals, moisturizing creams and lotions, and topical retinoids. Contact dermatitis from sorbitol derivatives appears to be increasingly prevalent. Patch-testing with SSO can be useful in the work-up of patients with presumptive cosmetic allergic contact dermatitis. Those sensitized to SSO can be counseled to avoid sorbitol containing products, especially topical corticosteroids. Herein we discuss case reports of SSO allergy and a recent case series of 12 of 112 dermatitis patients (10.7%) patch-tested during a 6-month period who showed contact allergy to SSO. We provide a list of key corticosteroids and products that can contain SSO, sorbitol, or sorbitol derivatives. PMID- 19134440 TI - Etiology of para-phenylenediamine sensitization: hair dye and other products. PMID- 19134441 TI - The comparative substantivity of three marking pens used in patch testing. PMID- 19134445 TI - Acupressure for chemotherapy-associated nausea and vomiting in children. AB - The objective of this study was to determine the feasibility and effectiveness of acupressure therapy in preventing chemotherapy-associated nausea in children. A prospective, randomized, crossover trial was conducted among pediatric oncology patients at Brenner Children's Hospital (Winston-Salem, NC). Patients were randomized to one of two treatment sequences involving acupressure wrist bands and placebo bands, separated by a standard care treatment with no bands. All patients received standard antiemetic therapy for each treatment. Expectations and outcomes of nausea and vomiting were assessed by questionnaires. Of 21 patients approached, 21 were enrolled and 18 completed all three study treatments. Patients' ages ranged from 5 to 19 years, 14 of 18 were Caucasian, and 9 were male. In general, patients expressed moderate expectations that acupressure would prevent nausea and vomiting. Following the session with an acupressure band, a third of all patients reported better than expected nausea prevention. There was no significant difference in nausea or vomiting between the three groups; there were no significant side effects from acupressure or placebo bands. Pediatric oncology patients have moderate expectations about the effectiveness of acupressure in preventing nausea and vomiting. Acupressure is feasible and well tolerated but was not more effective than placebo in this sample of patients who were also treated with standard antiemetic therapies. PMID- 19134446 TI - Parents' attitudes and expectations about music's impact on pediatric oncology patients. AB - Clinicians often have positive attitudes about the clinical effects of music. To better understand barriers to providing music in the clinic, we describe parents' attitudes about music for pediatric oncology outpatients. A cross-sectional survey was conducted between January 2005 and October 2007 in a pediatric oncology clinic in a tertiary hospital. Eligible subjects were one parent of pediatric leukemia patients. Surveys were distributed at a routine clinic visit as part of a study on the effects of music on subjective and objective well being. Of the 67 eligible families, 45 (67%) parents responded; 82% reported playing music for the patient at home within the previous week. The most common reasons to use music for the patient were to entertain (88%), keep the patient company (71%), help the patient feel better (76%), or provide comfort (69%); fewer used music to distract the patient from pain (16%) or nausea (11%). Parents expected that music during clinic visits would have positive effects: relaxation (64%), comfort (42%), and/or distraction (33%); none expected negative effects. Parents often play music for their children, and they hold favorable attitudes about playing it in the clinic. Parents' attitudes are not barriers to providing music in the clinic. PMID- 19134447 TI - Phytomedicine 101: plant taxonomy for preclinical and clinical medicinal plant researchers. AB - Plants are the primary source of medicine for most of the world. The most fundamental step in the scientific study of medicinal plants is establishing their botanical identity. Many studies lack voucher specimens, which serve as permanent records of scientific investigations. This omission makes positive identification impossible and hinders reproducibility. Even when vouchers are cited, scientific names are often mishandled. A random survey of titles and abstracts of 100 publications revealed 20 with taxonomic errors. Mistakes included a lack of author citations, misspellings, and use of older synonyms instead of currently accepted names. A seemingly minor orthographic error makes it impossible to search electronic databases for information about a species. Medicinal plant manuscripts and National Institutes of Health proposals commonly lack scientific rigor in dealing with botanical names and documentation. This article examines common taxonomic problems relevant to medicinal plant research and provides a basic guide to plant taxonomy for medicinal plant researchers. Voucher specimens and their preparation, plant identification, and botanical nomenclature are discussed. References and other resources to assist investigators are cited. PMID- 19134449 TI - Credentialing complementary practitioners in a large academic cancer center. AB - One of the key obstacles to the complete integration of complementary and integrative medicine (CIM) into standard care in the United States is the lack of between-state and between-institution standards for credentialing. Also, a formal framework for the scope of CIM practitioner's practice is not available for assessing CIM integration into conventional patient care. Although many cancer centers do have some CIM programming under way, the scope of practice for CIM practitioners who may or may not fall within any formal licensing body and for non-CIM practitioners continues to vary among centers. This variation can result in inconsistent outcomes, difficulties in educating cancer patients about the role CIM can play in their cancer care, and a lack of true integration of CIM therapies into conventional treatment planning for the patient and those who care about and for them. PMID- 19134450 TI - Red clover (Trifolium pratense). PMID- 19134448 TI - Breast cancer and the immune system. AB - This article reviews the evidence that the functioning of both the innate and the adaptive immune system plays a role in preventing relapse in women with breast cancer. Lymphocytes, including T cells, T regulatory cells, and natural killer cells, and their cytokine release patterns are implicated in both primary prevention and recurrence of breast cancer. Cancer prognosis may be related to immune system functional status. The hypothesis that the immune system has a causal role in breast cancer etiology is supported by epidemiologic, preclinical, and clinical research. Empirical support for the concept that immune status and immunomodulatory therapy have important roles in comprehensive breast cancer treatment is provided. PMID- 19134451 TI - [Prescription rules of Chinese herbal medicines in treatment of gastric cancer]. AB - Analysis of the nature, tastes, channel distributions and effects of the frequently used herbal medicines in the prescriptions involved in the clinical literatures about treatment of gastric cancer published from 1988 to 2007 was made in the paper. The literatures were categorized into three types: 1) treatment of middle- and late-stage gastric cancer; 2) prevention and treatment of the recurrence and metastasis after operation; 3) Chinese herbal medicines combined with chemotherapy for enhancing efficacy and reducing toxicity. The most frequently used herbal medicines in the three literature types were qi invigorating herbs, such as Atractylodes, Astragalus, Codonopsis, Glycyrrhiza and Ginseng, etc. The herbal medicines for promoting urination to subside swelling such as tuckahoe and Semen Coicis, etc were used more frequently than the herbal medicines for regulating qi such as dried orange peel and putchuck, etc, as well as for clearing away heat to remove toxin such as spreading hedyotis herb, Herba Scutellariae Barbatae, yangtao actinidia root, and Rhizoma Paridis, etc. From another angle, the most frequently used herbal medicines for the treatment of gatric cancer were those cold, warm and neutral in nature, sweet, bitter and pungent in taste, and distributed to spleen and liver channels. PMID- 19134452 TI - [Tanreqing Injection for community-acquired pneumonia: a systematic review of randomized evidence]. AB - OBJECTIVE: To evaluate the efficacy and safety of Tanreqing Injection, a compound traditional Chinese herbal medicine, for community-acquired pneumonia. METHODS: Literatures about randomized controlled trials of Tanreqing Injection for community-acquired pneumonia were reviewed. Related literatures were selected and analyzed according to different treatment strategies of the trials. The methodological quality of the trials was assessed by the Jadad scale, and evaluation was performed. RESULTS: Twelve randomized controlled trials meeting the inclusion criteria were selected and reviewed. As Tanreqing combined group (Tanreqing Injection plus antibiotics and basic therapy) was compared with antibiotics group (antibiotics plus basic therapy), the meta-analysis indicated that the relative risk (RR) for the total cure rate was 1.51, and 95% confidence interval (CI) was [1.29, 1.77]; RR for the total obvious effect rate was 1.31,ls and 95% CI was [1.20,1.43]; RR for the effective rate was 1.17, and 95% CI was [1.11, 1.23]. The weighted mean difference (WMD) in disappearance time of fever between the two groups was -1.24, and 95% CI was [-1.71, -0.76]. The RR values between the two groups for the total obvious effect rate of cough and expectoration were 1.42 and 1.27, and 95% CIs were [1.16, 1.74] and [1.04, 1.55] respectively. The RR values between the two groups in absorption of chest X-ray shadow and neutrophil number were 1.19, 1.10 and 95% CIs were [1.09, 1.30], [1.03, 1.17] respectively. The differences were all statistically significant. Serious systematic adverse reactions had not been reported in the trials. CONCLUSION: The effect of combined therapy with Tanreqing Injection plus antibiotics and basic therapy is better than that of antibiotics plus basic therapy. Tanreqing Injection can improve the symptoms of cough and expectoration, shorten the fever time and facilitate the absorption of chest X-ray shadow, without any significant adverse reactions. However, further high-quality trials are needed. PMID- 19134453 TI - [Development and evaluation of an inquiry scale for diagnosis of heart system syndromes in traditional Chinese medicine]. AB - OBJECTIVE: To develop an inquiry scale for diagnosis of heart system syndromes, and to discuss the provisional standardization of the inquiry method in traditional Chinese medicine (TCM). METHODS: Based on scale-making method, Chinese medicine theory and literature searching, an inquiry scale for diagnosis of heart system syndromes in TCM was developed. Statistics method, frequency counting and Delphi method were used for analysis. The inquiry scale was revised and tested repeatedly to check the test reliability, internal consistency reliability, and content validity, etc. RESULTS: The inquiry scale for diagnosis of heart system syndromes mainly covered basic data, chief complaint, history of present illness (accompanying symptoms) and past history, with appendix of inspection and palpation information as well as diagnosis made according to traditional Chinese and Western medicine. Among them, general inquiries covered fever and chills, sweating, head-body and chest-belly symptoms, taste and diet, stool and urine, sleep, mood, and gynecologic symptoms, which were scaled in 8 dimensions. And 66 symptom variables were screened finally. The scale had a good content validity and its coefficient alpha was 0.82. For the results of test retest reliability, the Kappa values of using the scale for diagnosis of heart-qi deficiency, heart-yang deficiency, turbid phlegm, and cold coagulation twice by the same doctor ranged from 0.74 to 1, showing that the consistency of the scale was relatively high. The Kappa values of evaluation of scorer reliability in diagnosis of heart-qi deficiency, heart-yang deficiency, and heart-yin deficiency were also high, which were 0.63, 0.72, 1 and 0.48 respectively. Other results of diagnosis had low-consistency or even no diagnostic agreement. CONCLUSION: The research on the scale for inquiry in TCM indicates that it is feasible for the standardization of inquiry scale for diagnosis of heart system syndromes in TCM, offering a reference for research on the inquiry scales for other systems. PMID- 19134454 TI - [Herbs for calming liver and suppressing liver-yang in treatment of migraine with hyperactive liver-yang syndrome and its effects on lymphocyte protein expression: a randomized controlled trial]. AB - OBJECTIVE: To observe the efficacy of herbs for calming liver and suppressing liver-yang in treatment of migraine patients with hyperactivity of liver-yang syndrome and to investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of pathogenic mechanisms in migraine with hyperactive liver-yang syndrome and the curative mechanisms of calming liver and suppressing liver-yang treatment. METHODS: A total of 32 migraine patients with hyperactivity of liver-yang syndrome were randomly divided into treatment group (16 cases) and control group (16 cases). The patients in the treatment group were treated with herbs for calming liver and suppressing liver-yang in accordance with traditional Chinese medicine (TCM) theory and the patients in the control group were treated with Flunarizine Capsules for two courses of treatment. The therapeutic effects, the score of TCM symptom and the changes of headache attack were observed in both groups before and after the treatment. The side effects were also observed in both groups. The level of differential protein expression was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). RESULTS: The herbs for calming liver and suppressing liver-yang had better effects on headache improvement than the Flunarizine Capsules (P<0.01). The cure rates in the treatment and control groups were 87.5% and 75.0% respectively. Vertigo, restlessness and tantrum, and prosopo heat in the treatment group were also improved as compared with those in the control group (P<0.05). After treatment, the score of TCM symptom in the two groups were all decreased (P<0.01), and there was a significant difference between the treatment group and the control group (P<0.01). The herbs for calming liver and suppressing liver-yang had no side effects. The average protein spots in the blood lymphocyte of normal people, migraine patients with hyperactivity of liver-yang syndrome in the treatment group before and after the treatment were (534+/-42), (552+/-54) and (529+/-55) spots respectively. Six down-regulated protein expressions and 14 up-regulated protein expressions were obtained in the treatment group. Four strengthened protein expressions in the six down-regulated proteins and 11 low protein expressions in the 14 up-regulated proteins were also obtained after treatment. Ten of the total 12 differential protein spots were successfully identified by MALDI-TOF-MS. The functions of these proteins were involved in metabolism, energy generation, transportation, antioxidation, signal transduction and immune, etc. According to information provided by NCBI and MSDB database, there were some proteins closely related to migraine with hyperactivity of liver-yang syndrome, such as peroxiredoxin 2, heat shock protein 27 and annexin A1. CONCLUSION: Herbs for calming liver and suppressing liver-yang is effective in treating migraine, and can improve TCM symptoms. The effects on migraine patients with hyperactivity of liver-yang syndrome may be related to regulating the blood lymphocyte protein expression. PMID- 19134455 TI - [Value of four scoring systems for predicting prognosis of severe acute pancreatitis]. AB - OBJECTIVE: To evaluate the value of Ranson, acute physiology and chronic health evaluation II (APACHE II), Balthazar CT and pancreatitis outcome prediction (POP) scoring systems in predicting the prognosis of severe acute pancreatitis (SAP). METHODS: A total of 253 consecutive patients with SAP, admitted into West China Hospital of Sichuan University from January 2004 to August 2007 were studied retrospectively. Data associated to Ranson, APACHE II, Balthazar CT and POP scoring systems after admissions were recorded. Four outcome signals including death, local complications, operation transformation, and development of multiple organ dysfunction syndrome (MODS) were analyzed. The receiver operating characteristic (ROC) curves were also drawn and the areas under the ROC curve (AUC) were obtained to compare the sensitivity, specificity, Youden index, and likelihood ratio positive test (LRPT) of the clinical scoring systems in assessing the prognosis of SAP. RESULTS: Through comparison of AUC, Ranson score could be used to predict the outcomes except for local complications (P=0.00), and all its diagnosis threshold was 5. APACHE II score showed that it could assess the outcomes of SAP except for local complications (P=0.00). Its thresholds in mortality, operation transformation, and incidence of MODS were 14, 10, and 12 respectively. Balthazar CT could predict all the outcomes, and the threshold was 6. POP score could predict all the outcomes except for local complications, and its predictive value in mortality was particularly prominent. The sensitivity and specificity of POP in predicting death were 0.90 and 0.92 as the cutoff was no less than 14. The thresholds of POP in mortality, operation transformation, and incidence of MODS were 14, 8, and 10 respectively. CONCLUSION: The prognostic ability of the four scoring systems is not the same. Ranson, APACHE II and POP scoring systems can predict the severity of SAP and entire complication, but they can't predict the local complications. POP score may predict the mortality well, and Balthazar CT scanning can predict all the outcomes, but the sensitivity and specificity are low. PMID- 19134456 TI - [Regulatory mechanism of malignant behavior of endometriosis mediated by puerarin]. AB - OBJECTIVE: To observe the inhibitory effects of puerarin on angiopoiesis of endometriotic tissue, and to explore the regulatory effects of puerarin on tumor related gene expression of endometriosis. METHODS: The regulatory effects of puerarin on endometriotic angiopoiesis and tumor-related gene expression were observed by using a chicken chorioallantoic membrane model and gene array method. RESULTS: Chicken chorioallantoic membrane experiment indicated that puerarin obviously inhibited endometriotic vesiculation and angiopoiesis. The area of blood vessels was significantly reduced as compared with the untreated group (P<0.05). The expressions of oncogenes and genes related to adhesion, invasion, and apoptosis, including ERBB2, ETS2, FOS, S100A4, TEK, TERT, NFKBIA, CDH1, CD44, ITGA6, NCAM1, MMP1, FLT1, AKT1, BCL2L and BIRC5 genes, were obviously higher, while the expressions of the anti-oncogenes, anti-apoptosis genes and anti invasion genes, including KAI1, KISS1, SERPINB5, TNFRSF25, TNFRSF1A, TNFRSF6 and SERPINB2, were significantly lower in eutopic endometrial tissue from patients with endometriosis than those from endometriosis-free women. The expressions of oncogenes (ERBB2, ETS2, FOS), apoptosis gene (BCL2L1), cyclin-dependent kinases (CDK4, CDC25A), and growth factor and receptors (HGF, FGFR2, TGFBR) were significantly enhanced, while the expressions of the anti-oncogenes (KAI1, SERPINB5), apoptosis genes (BAD and TNF) and cyclin-dependent kinase inhibiting factor (CDKN2A) were obviously reduced in ectopic tissue as compared with those in eutopic tissue from patients with endometriosis. Puerarin significantly enhanced the gene expressions in endometriotic stromal cells, including BAD, BAX, CASP8, CASP9, TNFRSF6, CDKN1B, CDKN2A, IFNA1 and IFNB1, and reduced the gene expressions of FOS, CHEK2, SRC, ITGB5, MMP9, PDGFA and NFKBIA. CONCLUSIONS: The tumor-related gene expression has significant differences in eutopic endometrial tissue between patients with endometriosis and endometriosis-free women, and between ectopic and eutopic tissues from patients with endometriosis. Puerarin can reduce angiopoiesis, regulate tumor-related gene expression and facilitate apoptosis in endometriotic tissue. PMID- 19134457 TI - [Protective effects of n-butanol extract of Potentilla anserina on acute myocardial ischemic injury in mice]. AB - OBJECTIVE: To investigate the protective effects of the n-butanol extract of Potentilla anserina L. (NP) on pituitrin-induced acute myocardial ischemic injury in mice. METHODS: Ninety healthy female mice were randomly divided into normal control group, untreated group, Salvia miltiorrhiza group and low-, medium- and high-dose NP groups. Except for the normal control group, the mice were intraperitoneally injected with pituitrin (20 U/kg) to induce acute myocardial ischemic injury. Thirty minutes after induction, electrocardiogram was monitored, and height of the J spot was measured also. Activities of lactate dehydrogenase (LDH), creatine kinase (CK) and superoxide dismutase (SOD) and content of malondialdehyde (MDA) in serum of the mice were detected. The degree of myocardial ischemic injury in mice was observed by Nagar-Olsen staining. RESULTS: The moving up of J spots in the treated groups was significantly inhibited when comparing with the untreated group (P<0.01). Compared with untreated group, high- and medium-dose NP and Salvia miltiorrhiza could significantly decrease the activities of LDH, CK (P<0.01, P<0.05), increase the SOD activity (P<0.01) and decrease the content of MDA (P<0.05). However, no significant difference was observed between low-dose NP group and untreated group (P>0.05). Nagar-Olsen staining showed that high- and medium-dose NP and Salvia miltiorrhiza could significantly diminish the areas of cardiac muscles injured by ischemia, but low dose NP had no effect on that. CONCLUSION: NP has a remarkable protective effect on acute myocardial ischemic injury in mice. PMID- 19134458 TI - [Mechanism of Yiqi Huayu Bushen recipe in treating cervical syndrome in rats with qi deficiency, blood stasis and kidney deficiency]. AB - OBJECTIVE: To explore the mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound traditional Chinese herbal medicine, in treating cervical syndrome (CS) with qi deficiency, blood stasis and kidney deficiency in rats. METHODS: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, untreated group and YHBR group; there were ten rats in each group. The rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency was established by combining disease and syndrome models. After one-month YHBR treatment in YHBR group, all rats were sacrificed. Serum, plasma and cervical intervertebral discs were detected and observed by radio-immunology, histopathology, immunohistochemistry, etc. RESULTS: Compared with those in the normal control group, rats in the untreated group showed obvious signs of deficiency in vital energy, such as tiredness, ptosis, few movement, bluish purple tongue and tail and weight loss; the weight of uterus and appendages, and the ratio of cyclic adenosine monophosphate/cyclic guanosine monophosphate were decreased; hemorheological parameters and the expression of alpha-granular membrane protein (CD62p) were increased; the content of serum estradiol was decreased. YHBR could improve the above indexes except for the weight of uterus and appendages. HE staining showed annulus fibrosus with cracks, diminish of pulposus nucleus, and decrease in the height of intervertebral disc and the thickness of end-plate in the untreated group; and YHBR could improve the changes of cervical intervertebral discs, but no obvious changes in end-plate. Type II collagen protein (Col2a1)was distributed in every part of the annulus fibrosus in normal control group, but was decreased in untreated group; YHBR could increase Col2a1 expression in annulus fibrosus as compared with the untreated group. The expressions of Col2a1 and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs were decreased in untreated group as compared with the normal control group (P<0.01); the expressions of Col2a1 and TIMP-1 mRNAs were higher and matrix metalloproteinase-13 mRNA was lower in the YHBR group than those in the untreated group (P<0.05, P<0.01). CONCLUSION: YHBR may improve the condition of cervical syndrome with qi deficiency, blood stasis and kidney deficiency by regulating immune system, coagulation system and endocrine system, and delay the degeneration of cervical intervertebral disc by regulating extracellular matrix and metalloproteases in intervertebral disc. PMID- 19134459 TI - [Interventional effects of emodin on transforming growth factor-beta1/integrin linked kinase signal way in interleukin-1beta-induced transdifferentiation of rat tubular epithelial-myofibroblasts]. AB - OBJECTIVE: To study the effects of transforming growth factor-beta1/integrin linked kinase (TGF-beta1/ILK) signal way in interleukin-1beta (IL-1beta)-induced rat tubular epithelial-myofibroblast transdifferentiation (TEMT), and to investigate whether emodin inhibits IL-1beta-induced TEMT through the TGF beta1/ILK signal way-dependent mechanism. METHODS: Normal rat kidney epithelial cell line (NRK52E) was used in this study. NRK52E cells were divided into blank control group, emodin control group, IL-1beta-induced group, emodin-inhibited group, SB431542 (TGF-beta 1 type I receptor blocker)-inhibited group, emodin plus SB431542-inhibited group, emodin-pretreated group and emodin-reversed group. After 48-hour culture, morphological changes of the NRK52E cells were observed by an inverted phase contrast microscope. The expressions of alpha-smooth muscle actin (alpha-SMA) and E-cadherin were detected by two-color immunohistochemical staining, while the expressions of TGF-beta1 and ILK were detected by one-color immunohistochemical staining. We also performed the imaging analysis to quantitatively analyze the result of the immunohistochemical staining. The secretion of fibronectin (FN) was analyzed by enzyme-linked immunosorbent assay. RESULTS: Compared with the blank control group, IL-1beta might induce TEMT, which was showed in increasing expression of alpha-SMA, increasing secreting of FN and decreasing expression of E-cadherin, and at the same time the expressions of TGF beta1 and ILK were enhanced (P<0.05). Emodin might inhibit all of those changes induced by IL-1beta (P<0.05). When TGF-beta1 signal way was intercepted, IL-1beta induced-TEMT was suppressed and the expression of ILK was decreased, however, there was no significant difference in expression of TGF-beta1 between the SB431542 group and the IL-1beta-induced group. Compared with emodin-inhibited group, emodin-pretreatment could not prevent IL-1beta induced-TEMT in a certain extent, but emodin could not revert IL-1beta-induced TEMT. Spearman correlation analysis showed that TGF-beta1 expression had positive correlation with expressions of alpha-SMA, FN, ILK and negative correlation with E-cadherin expression, and the expression of ILK was positively correlated with the expressions of alpha-SMA and FN and negatively correlated with E-cadherin expression. CONCLUSION: IL-1beta induces TEMT partly depending on TGF-beta1/ILK signal way, partly via which emodin inhibits the TEMT induced by IL-1beta. PMID- 19134460 TI - [Effects of ligustrazine injection on high glucose-induced type I collagen, matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 expressions in human peritoneal mesothelial cells in vitro]. AB - OBJECTIVE: To investigate the effects of ligustrazine injection on type I collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) expressions in human peritoneal mesothelial cells (HPMCs) cultured in high glucose conditions. METHODS: HPMCs were isolated from human omenta by trypsin digestion method and subcultured. Then, the HPMCs were divided into normal control group, high glucose group and high glucose plus low-, medium- and high-dose ligustrazine (10, 20 and 40 mg/L ligustrazine respectively) groups. Semi-quantitative reverse transcription-polymerase chain reaction was used to detect the expressions of type I collagen, MMP-1 and TIMP-1 mRNAs in HPMCs. Proteins of type I collagen, MMP-1 and TIMP-1 in culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Cell protein concentration was measured by trace bicinchoninic acid method to correct the ELISA assay results. RESULTS: Ligustrazine injection could significantly decrease high glucose-induced type I collagen and TIMP-1 expressions in a dose-dependent manner both in protein and gene levels (P<0.05, P<0.01). In addition, medium- and high dose ligustrazine injection could significantly increase MMP-1 expression which was inhibited by high glucose concentrations (P<0.05). CONCLUSION: Ligustrazine injection does not only decrease type I collagen synthesis, but also promote its degradation by modulating unbalanced MMP-1/TIMP-1 expression in HPMCs cultured in high glucose conditions. PMID- 19134461 TI - [Research advances in interior-exterior relationships between zang-organs and fu organs]. PMID- 19134462 TI - [How to identify statistical data]. PMID- 19134463 TI - Comparative study on WHO Western Pacific Region and World Federation of Chinese Medicine Societies international standard terminologies on traditional medicine: an analysis of the Body Constituents. PMID- 19134464 TI - [English translation of antithetical words in Huangdi Neijing]. PMID- 19134465 TI - [Application of complementary and alternative therapies abroad in treatment of climacteric symptoms]. PMID- 19134466 TI - The impact of seasonal variability in wildlife populations on the predicted spread of foot and mouth disease. AB - Modeling potential disease spread in wildlife populations is important for predicting, responding to and recovering from a foreign animal disease incursion such as foot and mouth disease (FMD). We conducted a series of simulation experiments to determine how seasonal estimates of the spatial distribution of white-tailed deer impact the predicted magnitude and distribution of potential FMD outbreaks. Outbreaks were simulated in a study area comprising two distinct ecoregions in South Texas, USA, using a susceptible-latent-infectious-resistant geographic automata model (Sirca). Seasonal deer distributions were estimated by spatial autoregressive lag models and the normalized difference vegetation index. Significant (P < 0.0001) differences in both the median predicted number of deer infected and number of herds infected were found both between seasons and between ecoregions. Larger outbreaks occurred in winter within the higher deer-density ecoregion, whereas larger outbreaks occurred in summer and fall within the lower deer-density ecoregion. Results of this simulation study suggest that the outcome of an FMD incursion in a population of wildlife would depend on the density of the population infected and when during the year the incursion occurs. It is likely that such effects would be seen for FMD incursions in other regions and countries, and for other diseases, in cases in which a potential wildlife reservoir exists. Study findings indicate that the design of a mitigation strategy needs to take into account population and seasonal characteristics. PMID- 19134467 TI - In vivo labeling method using a genetic construct for nanoscale resolution microscopy. AB - We demonstrate beam scanning-stimulated emission depletion microscopy with in vivo labeled cells. A red emitting fluorescent dye is introduced into membrane protein fused to a multifunctional reporter protein (HaloTag, Promega, Madison, WI) in live cells. This approach allows superresolution stimulated emission depletion imaging without the limitations of immunofluorescence-based staining. PMID- 19134468 TI - Simultaneous measurement of nucleotide occupancy and mechanical displacement in myosin-V, a processive molecular motor. AB - Adenosine triphosphate (ATP) turnover drives various processive molecular motors and adenosine diphosphate (ADP) release is a principal transition in this cycle. Biochemical and single molecule mechanical studies have led to a model in which a slow ADP release step contributes to the processivity of myosin-V. To test the relationship between force generation and ADP release, we utilized optical trapping nanometry and single molecule total internal reflection fluorescence imaging for simultaneous and direct observation of both processes in myosin-V. We found that ADP was released 69 +/- 5.3 ms after force generation and displacement of actin, providing direct evidence for slow ADP release. As proposed by several previous studies, this slow ADP release probably ensures processivity by prolonging the strong actomyosin state in the ATP turnover cycle. PMID- 19134469 TI - A CaV1.1 Ca2+ channel splice variant with high conductance and voltage sensitivity alters EC coupling in developing skeletal muscle. AB - The Ca(2+) channel alpha(1S) subunit (Ca(V)1.1) is the voltage sensor in skeletal muscle excitation-contraction (EC) coupling. Upon membrane depolarization, this sensor rapidly triggers Ca(2+) release from internal stores and conducts a slowly activating Ca(2+) current. However, this Ca(2+) current is not essential for skeletal muscle EC coupling. Here, we identified a Ca(V)1.1 splice variant with greatly distinct current properties. The variant of the CACNA1S gene lacking exon 29 was expressed at low levels in differentiated human and mouse muscle, and up to 80% in myotubes. To test its biophysical properties, we deleted exon 29 in a green fluorescent protein (GFP)-tagged alpha(1S) subunit and expressed it in dysgenic (alpha(1S)-null) myotubes. GFP-alpha(1S)Delta 29 was correctly targeted into triads and supported skeletal muscle EC coupling. However, the Ca(2+) currents through GFP-alpha(1S)Delta 29 showed a 30-mV left-shifted voltage dependence of activation and a substantially increased open probability, giving rise to an eightfold increased current density. This robust Ca(2+) influx contributed substantially to the depolarization-induced Ca(2+) transient that triggers contraction. Moreover, deletion of exon 29 accelerated current kinetics independent of the auxiliary alpha(2)delta-1 subunit. Thus, characterizing the Ca(V)1.1 Delta 29 splice variant revealed the structural bases underlying the specific gating properties of skeletal muscle Ca(2+) channels, and it suggests the existence of a distinct mode of EC coupling in developing muscle. PMID- 19134470 TI - Structural rearrangements in loop F of the GABA receptor signal ligand binding, not channel activation. AB - Structure-function studies of the Cys loop family of ionotropic neurotransmitter receptors (GABA, nACh, 5-HT(3), and glycine receptors) have resulted in a six loop (A-F) model of the agonist-binding site. Key amino acids have been identified in these loops that associate with, and stabilize, bound ligand. The next step is to identify the structural rearrangements that couple agonist binding to channel opening. Loop F has been proposed to move upon receptor activation, although it is not known whether this movement is along the conformational pathway for channel opening. We test this hypothesis in the GABA receptor using simultaneous electrophysiology and site-directed fluorescence spectroscopy. The latter method reveals structural rearrangements by reporting changes in hydrophobicity around an environmentally sensitive fluorophore attached to defined positions of loop F. Using a series of ligands that span the range from full activation to full antagonism, we show there is no correlation between the rearrangements in loop F and channel opening. Based on these data and agonist docking simulations into a structural model of the GABA binding site, we propose that loop F is not along the pathway for channel opening, but rather is a component of the structural machinery that locks ligand into the agonist-binding site. PMID- 19134471 TI - Diffusion, exclusion, and specific binding in a large channel: a study of OmpF selectivity inversion. AB - We find that moderate cationic selectivity of the general bacterial porin OmpF in sodium and potassium chloride solutions is inversed to anionic selectivity in concentrated solutions of barium, calcium, nickel, and magnesium chlorides. To understand the origin of this phenomenon, we consider several factors, which include the binding of divalent cations, electrostatic and steric exclusion of differently charged and differently sized ions, size-dependent hydrodynamic hindrance, electrokinetic effects, and significant "anionic" diffusion potential for bulk solutions of chlorides of divalent cations. Though all these factors contribute to the measured selectivity of this large channel, the observed selectivity inversion is mostly due to the following two. First, binding divalent cations compensates, or even slightly overcompensates, for the negative charge of the OmpF protein, which is known to be the main cause of cationic selectivity in sodium and potassium chloride solutions. Second, the higher anionic (versus cationic) transport rate expected for bulk solutions of chloride salts of divalent cations is the leading cause of the measured anionic selectivity of the channel. Interestingly, at high concentrations the binding of cations does not show any pronounced specificity within the divalent series because the reversal potentials measured in the series correlate well with the corresponding bulk diffusion potentials. Thus our study shows that, in contrast to the highly selective channels of neurophysiology that employ mostly the exclusion mechanism, quite different factors account for the selectivity of large channels. The elucidation of these factors is essential for understanding large channel selectivity and its regulation in vivo. PMID- 19134472 TI - Magainin 2 revisited: a test of the quantitative model for the all-or-none permeabilization of phospholipid vesicles. AB - The all-or-none kinetic model that we recently proposed for the antimicrobial peptide cecropin A is tested here for magainin 2. In mixtures of phosphatidylcholine (PC)/phosphatidylglycerol (PG) 50:50 and 70:30, release of contents from lipid vesicles occurs in an all-or-none fashion and the differences between PC/PG 50:50 and 70:30 can be ascribed mainly to differences in binding, which was determined independently and is approximately 20 times greater to PC/PG 50:50 than to 70:30. Only one variable parameter, beta, corresponding to the ratio of the rates of pore opening to pore closing, is used to fit dye release kinetics from these two mixtures, for several peptide/lipid ratios ranging from 1:25 to 1:200. However, unlike for cecropin A where it stays almost constant, beta increases five times as the PG content of the vesicles increases from 30 to 50%. Thus, magainin 2 is more sensitive to anionic lipid content than cecropin A. But overall, magainin follows the same all-or-none kinetic model as cecropin A in these lipid mixtures, with slightly different parameter values. When the PG content is reduced to 20 mol %, dye release becomes very low; the mechanism appears to change, and is consistent with a graded kinetic model. We suggest that the peptide may be inducing formation of PG domains. In either mechanism, no peptide oligomerization occurs and magainin catalyzes dye release in proportion to its concentration on the membrane in a peptide state that we call a pore. We envision this structure as a chaotic or stochastic type of pore, involving both lipids and peptides, not a well-defined, peptide-lined channel. PMID- 19134473 TI - Monitoring of an RNA multistep folding pathway by isothermal titration calorimetry. AB - Isothermal titration calorimetry was used to monitor the energetic landscape of a catalytic RNA, specifically that of the hepatitis delta virus ribozyme. Using mutants that isolated various tertiary interactions, the thermodynamic parameters of several ribozyme-substrate intermediates were determined. The results shed light on the impact of several tertiary interactions on the global structure of the ribozyme. In addition, the data indicate that the formation of the P1.1 pseudoknot is the limiting step of the molecular mechanism. Last, as illustrated here, isothermal titration calorimetry appears to be a method of choice for the elucidation of an RNA's folding pathway. PMID- 19134474 TI - Induced secondary structure and polymorphism in an intrinsically disordered structural linker of the CNS: solid-state NMR and FTIR spectroscopy of myelin basic protein bound to actin. AB - The 18.5 kDa isoform of myelin basic protein (MBP) is a peripheral membrane protein that maintains the structural integrity of the myelin sheath of the central nervous system by conjoining the cytoplasmic leaflets of oligodendrocytes and by linking the myelin membrane to the underlying cytoskeleton whose assembly it strongly promotes. It is a multifunctional, intrinsically disordered protein that behaves primarily as a structural stabilizer, but with elements of a transient or induced secondary structure that represent binding sites for calmodulin or SH3-domain-containing proteins, inter alia. In this study we used solid-state NMR (SSNMR) and Fourier transform infrared (FTIR) spectroscopy to study the conformation of 18.5 kDa MBP in association with actin microfilaments and bundles. FTIR spectroscopy of fully (13)C,(15)N-labeled MBP complexed with unlabeled F-actin showed induced folding of both protein partners, viz., some increase in beta-sheet content in actin, and increases in both alpha-helix and beta-sheet content in MBP, albeit with considerable extended structure remaining. Solid-state NMR spectroscopy revealed that MBP in MBP-actin assemblies is structurally heterogeneous but gains ordered secondary structure elements (both alpha-helical and beta-sheet), particularly in the terminal fragments and in a central immunodominant epitope. The overall conformational polymorphism of MBP is consistent with its in vivo roles as both a linker (membranes and cytoskeleton) and a putative signaling hub. PMID- 19134475 TI - Reconstitution of an actin cortex inside a liposome. AB - The composite and versatile structure of the cytoskeleton confers complex mechanical properties on cells. Actin filaments sustain the cell membrane and their dynamics insure cell shape changes. For example, the lamellipodium moves by actin polymerization, a mechanism that has been studied using simplified experimental systems. Much less is known about the actin cortex, a shell-like structure underneath the membrane that contracts for cell movement. We have designed an experimental system that mimicks the cell cortex by allowing actin polymerization to nucleate and assemble at the inner membrane of a liposome. Actin shell growth can be triggered inside the liposome, which offers a useful system for a controlled study. The observed actin shell thickness and estimated mesh size of the actin structure are in good agreement with cellular data. Such a system paves the way for a thorough characterization of cortical dynamics and mechanics. PMID- 19134476 TI - Multistep aggregation pathway of human interleukin-1 receptor antagonist: kinetic, structural, and morphological characterization. AB - The complex, multistep aggregation kinetic and structural behavior of human recombinant interleukin-1 receptor antagonist (IL-1ra) was revealed and characterized by spectral probes and techniques. At a certain range of protein concentration (12-27 mg/mL) and temperature (44-48 degrees C), two sequential aggregation kinetic transitions emerge, where the second transition is preceded by a lag phase and is associated with the main portion of the aggregated protein. Each kinetic transition is linked to a different type of aggregate population, referred to as type I and type II. The aggregate populations, isolated at a series of time points and analyzed by Fourier-transform infrared spectroscopy, show consecutive protein structural changes, from intramolecular (type I) to intermolecular (type II) beta-sheet formation. The early type I protein spectral change resembles that seen for IL-1ra in the crystalline state. Moreover, Fourier transform infrared data demonstrate that type I protein assembly alone can undergo a structural rearrangement and, consequently, convert to the type II aggregate. The aggregated protein structural changes are accompanied by the aggregate morphological changes, leading to a well-defined population of interacting spheres, as detected by scanning electron microscopy. A nucleation driven IL-1ra aggregation pathway is proposed, and assumes two major activation energy barriers, where the second barrier is associated with the type I --> type II aggregate structural rearrangement that, in turn, serves as a pseudonucleus triggering the second kinetic event. PMID- 19134477 TI - Backbone structure of transmembrane domain IX of the Na+/proline transporter PutP of Escherichia coli. AB - The backbone structure is determined by site-directed spin labeling, double electron electron resonance measurements of distances, and modeling in terms of a helix-loop-helix construct for a transmembrane domain that is supposed to line the translocation pathway in the 54.3 kDa Na(+)/proline symporter PutP of Escherichia coli. The conformational distribution of the spin labels is accounted for by a rotamer library. An ensemble of backbone models with a root mean-square deviation of less than 2 A is obtained. These models exhibit a pronounced kink near residue T341, which is involved in substrate binding. The kink may be associated with a hinge that allows the protein to open and close an inwardly oriented cavity. PMID- 19134478 TI - Single-molecule imaging of a fluorescent unnatural amino acid incorporated into nicotinic receptors. AB - We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine receptor, nAChR) in a living cell. Xenopus oocytes were injected with a frameshift-suppressor tRNA, amino-acylated with Lys(BODIPYFL) and nAChR (alpha/beta19'GGGU/gamma/delta) mRNAs. We measured fluorescence from oocytes expressing nAChR beta19'Lys(BODIPYFL), using time resolved total internal reflection fluorescence microscopy. Under conditions of relatively low receptor density (<0.1 receptors/microm(2)), we observed puncta with diffraction-limited profiles that were consistent with the point-spread function of our microscope. Furthermore, diffraction-limited puncta displayed step decreases in fluorescence intensity, consistent with single-molecule photobleaching. The puncta densities agreed with macroscopic ACh-induced current densities, showing that the fUAA was incorporated, and that receptors were functional. Dose-response relations for the nAChR beta19'Lys(BODIPYFL) receptors were similar to those for wild-type receptors. We also studied nAChR beta19'Lys(BODIPYFL) receptors labeled with alpha-bungarotoxin monoconjugated with Alexa488 (alphaBtxAlexa488). The nAChR has two alphaBtx binding sites, and puncta containing the Lys(BODIPYFL) labeled with alphaBtxAlexa488 yielded the expected three discrete photobleaching steps. We also performed positive control experiments with a nAChR containing enhanced green fluorescent protein in the gamma-subunit M3-M4 loop, which confirmed our nAChR beta19'Lys(BODIPYFL) measurements. Thus, we report on the cell-based single-molecule detection of nAChR beta19'Lys(BODIPYFL). PMID- 19134479 TI - Active site formation, not bond kinetics, limits adhesion rate between human neutrophils and immobilized vascular cell adhesion molecule 1. AB - The formation of receptor ligand bonds at the interface between different cells and between cells and substrates is a widespread phenomenon in biological systems. Physical measurements of bond formation rates between cells and substrates have been exploited to increase our understanding of the biophysical mechanisms that regulate bond formation at interfaces. Heretofore, these measurements have been interpreted in terms of simple bimolecular reaction kinetics. Discrepancies between this simple framework and the behavior of neutrophils adhering to surfaces expressing vascular cell adhesion molecule 1 (VCAM-1) motivated the development of a new kinetic framework in which the explicit formation of active bond formation sites (reaction zones) are a prerequisite for bond formation to occur. Measurements of cells interacting with surfaces having a wide range of VCAM-1 concentrations, and for different durations of contact, enabled the determination of novel kinetic rate constants for the formation of reaction zones and for the intrinsic bond kinetics. Comparison of these rates with rates determined previously for other receptor ligand pairs points to a predominant role of extrinsic factors such as surface topography and accessibility of active molecules to regions of close contact in determining forward rates of bond formation at cell interfaces. PMID- 19134483 TI - The challenge is to adopt the best of the new without diminishing scientific quality. PMID- 19134480 TI - Adhesion between human neutrophils and immobilized endothelial ligand vascular cell adhesion molecule 1: divalent ion effects. AB - Integrin-mediated adhesion of circulating neutrophils to endothelium during inflammation involves multiple adhesion molecules on both neutrophils and endothelium. Most studies of neutrophil adhesion have focused on adhesion to ICAM 1 (mediated by beta(2) integrins), but interaction with the endothelial ligand vascular cell adhesion molecule 1 (VCAM-1) may also play a role in neutrophil adhesion to activated endothelium. In this study we demonstrate significant adhesion between neutrophils and VCAM-1 mediated by beta(1) integrins, principally via alpha(4)beta(1) (VLA-4). We characterize the dynamics of adhesion in terms of rate constants for a two-step bond formation process, the first involving juxtaposition of active molecules with substrate and the second involving bond formation. The results indicate that the first step is rate limiting for VLA-4-VCAM-1 interactions. Changing divalent cation composition affects these coefficients, implicating molecular conformational changes as a key step in the process. PMID- 19134481 TI - The transfer functions of cardiac tissue during stochastic pacing. AB - The restitution properties of cardiac action potential duration (APD) and conduction velocity (CV) are important factors in arrhythmogenesis. They determine alternans, wavebreak, and the patterns of reentrant arrhythmias. We developed a novel approach to characterize restitution using transfer functions. Transfer functions relate an input and an output quantity in terms of gain and phase shift in the complex frequency domain. We derived an analytical expression for the transfer function of interbeat intervals (IBIs) during conduction from one site (input) to another site downstream (output). Transfer functions can be efficiently obtained using a stochastic pacing protocol. Using simulations of conduction and extracellular mapping of strands of neonatal rat ventricular myocytes, we show that transfer functions permit the quantification of APD and CV restitution slopes when it is difficult to measure APD directly. We find that the normally positive CV restitution slope attenuates IBI variations. In contrast, a negative CV restitution slope (induced by decreasing extracellular [K(+)]) amplifies IBI variations with a maximum at the frequency of alternans. Hence, it potentiates alternans and renders conduction unstable, even in the absence of APD restitution. Thus, stochastic pacing and transfer function analysis represent a powerful strategy to evaluate restitution and the stability of conduction. PMID- 19134484 TI - A surgeon now board certified in palliative care. PMID- 19134486 TI - Principles and core competencies of surgical palliative care: an overview. AB - The concept of palliation is as old as surgery itself, perhaps so old that it has been taken for granted rather than conceptualized as a primary framework for surgical care. The experience and success of the hospice movement in the United States and abroad was followed by the extension of its basic concepts to the much larger population of patients with advanced, but not necessarily terminal, illness. This collective experience has provided the necessary background and stimulus for developing a specific set of principles and competencies applicable to surgical palliative care. Surgical palliative care is the treatment of suffering and the promotion of quality of life for seriously or terminally ill patients under surgical care. PMID- 19134487 TI - Subjective idiopathic tinnitus and palliative care: a plan for diagnosis and treatment. AB - This article integrates the highlights of the authors' clinical experiences derived from existing protocols for tinnitus diagnosis and treatment with the evolving discipline of palliation medicine. Specifically, it demonstrates how the inclusion of principles of palliation medicine contributes to the efficacy of treatment. PMID- 19134488 TI - Palliative care for the patient with refractory chronic rhinosinusitis. AB - Most otolaryngologists encounter patients with chronic rhinosinusitis who, despite conventional medical and surgical therapy, fail to show significant symptomatic improvement. Many paradigms have been proposed to explain the mechanisms responsible for refractory disease in these patients, including superantigen activation, biofilm formation, and eosinophil activation triggered by fungal elements. Although the precise underlying etiology of this clinical scenario remains unclear, the resultant pathophysiologic events share a final common pathway marked by inflammatory changes of the sinonasal mucosa. This article reviews the proposed hypotheses as to why some patients with chronic sinusitis fail conventional therapy and highlights treatment options useful in the palliative treatment of these patients. PMID- 19134489 TI - Rehabilitation after cranial base surgery. AB - Recent advancements in skull base surgery to remove or diminish the size of cranial base tumors allow more to be done than ever before to preserve life for patients who have tumors in anatomic locations once considered unreachable without causing massive functional impairment or death. Nonetheless, the resulting outcome has a direct and serious impact on the quality of life of the patient. In this article on palliation, the authors focus on the rehabilitative techniques used in patients who have undergone extensive cranial base resection. These techniques can also be used to improve the life of patients who have not undergone surgery but suffer from poor quality of life because of the natural growth of the tumor. PMID- 19134490 TI - Palliative aspects of recurrent respiratory papillomatosis. AB - Recurrent respiratory papillomatosis (RRP) is a chronic, frequently debilitating, and potentially life-threatening disease. Therapy for RRP has evolved from simply inserting a tracheotomy to provide an airway and plucking out papillomata with cup forceps to provide some degree of voice to the present-day far more sophisticated approaches, along with preventative measures that may someday offer the potential dramatically to decrease disease prevalence. Family dynamics and support and intentional structuring of office protocols to accommodate the unique nature of RRP are as essential as any operative intervention for saving and prolonging life. This article reviews recent developments in the management of RRP and highlights palliative approaches to case management for those patients who are not easily cured with initial endoscopic interventions. PMID- 19134491 TI - Chronic subjective dizziness. AB - This article reviews the authors' work, which expands on previous studies to confirm that anxiety-related processes cause or maintain symptoms of dizziness. Discussed are interventions directed at patients' underlying psychologic disorders, including current methods of pharmacotherapy and psychotherapy. Patients with chronic complaints of nonspecific dizziness can present frustrating diagnostic and therapeutic challenges, but can be offered definitive and palliative care. The authors emphasize the importance of eliciting a precise description of the dizziness sensation from the patient as the critical factor in delineating the specific diagnosis and guiding treatment. PMID- 19134492 TI - Hearing loss. AB - For physicians treating patients with sensorineural hearing loss, therapy is directed more toward helping the patient cope with the loss of hearing rather than offering various medical or surgical interventions. Accordingly, for the patient with sensorineural hearing loss, the care plan is usually more directed toward palliation than toward cure. This article views hearing loss not only as a physiologic deficit, but as the loss of an important aspect of overall communication skill that can have far reaching emotional and psychologic effects on the patient, the family, and those who surround patients in their daily lives. In this article the authors offer strategies for managing the patient who is losing or who has lost hearing. PMID- 19134493 TI - Disorders of swallowing: palliative care. AB - This article defines palliative care for swallowing disorders as treatment for severe and chronic dysphagia or intractable aspiration when the recovery of normal swallowing is not anticipated and attempts to restore normal swallowing have been unsuccessful. Palliative treatment for dysphagia is not only for the dying patient because patients with difficulty swallowing can live for a long time. Palliative care for dysphagia is aimed at maximizing swallowing function, maintaining pulmonary health, and supporting healthy nutrition despite the impaired ability to swallow. When despite all attempts at intervention a patient becomes totally unable to swallow, the goal of therapy changes toward finding ways to provide adequate nutrition for the patient. PMID- 19134494 TI - Palliative treatment of dysphonia and dysarthria. AB - The focus of this article is the palliative treatment of a variety of dysphonic conditions. Symptomatic relief of hoarseness can be achieved by voice therapy, augmentative alternative communication modalities, and surgery. The causes of dysphonia addressed herein include amyotrophic lateral sclerosis, Parkinson's disease, multiple sclerosis, stroke, head and neck cancers requiring glossectomy or laryngectomy, unilateral vocal fold paralysis, and presbyphonia. Palliative treatment of dysphonia and voice disorders provides symptomatic relief but not a cure of the underlying disease state. For these patients there are a number of palliative interventions that can greatly improve their quality of life. PMID- 19134495 TI - Anosmia: loss of smell in the elderly. AB - More than 35 million people in the United States are aged 65 years or older. Of people aged more than 80 years, 62% to 80% have a significant reduction in their sense of smell, which can adversely affect their safety and nutritional status. This article reviews the anatomy of smell and discusses the known and theorized etiologies of smell loss in the elderly population. PMID- 19134496 TI - Tracheostomy in palliative care. AB - The use of tracheostomy in palliative care offers a viable option for airway control. Through a dialogue with the patient, family, and a multidisciplinary set of providers, this procedure can be a useful component to a patient's overall palliative care plan. The authors discuss techniques for placing a tracheostomy tube and indications for placement in palliative care for alleviation of chronic and acute symptoms. PMID- 19134497 TI - Alleviating head and neck pain. AB - Pain in patients who have cancer can be caused by direct effects of the tumor or by complications of treatment, or it can be unrelated to the disease or its treatment. This article discusses interventional pain procedures in the treatment of head and neck pain as they relate to malignancies and cervicogenic headache and neck pain. PMID- 19134498 TI - The role of vestibular rehabilitation in the balance disorder patient. AB - The focus of this article is on the role of vestibular rehabilitation in the remediation of postural and gaze instability in individuals with peripheral vestibular deficits. Although vestibular rehabilitation does not cure the organic disease that produces the balance disorder, it improves mobility, prevents falls, and overall has a positive impact on the quality of life for the patient. PMID- 19134499 TI - Living with head and neck cancer and coping with dying when treatments fail. AB - Palliative care in patients who have head and neck cancer is a complex topic that requires a multifaceted approach. The head and neck surgeon has an important duty to fulfill in managing and following the wishes of the incurable cancer patient and is obligated to direct them to the appropriate services in this challenging time. PMID- 19134500 TI - Coronary artery pseudotumor: IgG4-related disease versus localized Castleman disease. PMID- 19134502 TI - The impact of obesity on health care costs among persons with schizophrenia. AB - BACKGROUND: Obesity is the second leading cause of preventable death in the United States and is twice as common among individuals with schizophrenia as the general population. METHODS: Data from the Clinical Antipsychotic Trials of Intervention Effectiveness, a multisite trial of antipsychotic pharmacotherapy in 1460 patients with schizophrenia, were used to examine the relationships between body mass index (BMI) and medical costs. RESULTS: ANCOVA analyses found significant increases in both psychiatric and nonpsychiatric medication costs associated with increasing BMI and a significant, but smaller, difference in costs of outpatient medical-surgical service utilization: US$41 per month for morbidly obese patients compared to US$26 per month for patients of normal weight (F=2.4, P=.04). In multivariable logistic regression analyses, morbid obesity was associated with significantly increased odds of any outpatient medical-surgical service costs. When compared to observations of BMI>35, BMI observations within the normal range (18.5-24.9) were half as likely to be associated with any outpatient medical-surgical costs (OR=0.53; 95% CI=0.45, 0.63). CONCLUSIONS: In this large sample of persons with schizophrenia, obesity was associated with increased outpatient general medical service and medication costs even after controlling for demographic characteristics and medical comorbidity, but the absolute dollar amount was small. PMID- 19134503 TI - Single item on positive affect is associated with 1-year survival in consecutive medical inpatients. AB - OBJECTIVE: To determine the independent effects of positive and negative affect items on mortality in consecutive medical inpatients. METHODS: Consecutive general medical inpatients were asked to complete the Hospital Anxiety and Depression Scale (HADS) at admission. Prognostic indicators were obtained from patients' records and physicians' ratings. The study end point was mortality from all causes at 1 year. RESULTS: The baseline assessment was completed by 575 patients (87.7%). Survival data were available for 572 of these (86 deaths). HADS depression scores and several physical risk indicators predicted mortality. Independent effects could be observed for HADS item 1 ("Can enjoy things as much as before") adjusted for physicians' ratings of prognosis, a principal diagnosis of hemato-oncological disease and Charlson comorbidity scores. In contrast, HADS depression items 2-7 (Model 1) as well as positive HADS depression scores did not contribute significantly to the prediction of mortality. CONCLUSION: Our present results suggest that one single item on positive affect independently predicts 1 year survival in consecutively admitted medical inpatients. Interestingly, this item has a stronger association with survival status than the presence of depressed mood. PMID- 19134504 TI - The relationship between migraine and mental disorders in a population-based sample. AB - OBJECTIVE: There is emerging evidence from clinical and community samples to suggest that migraines are associated with mental disorders. The present study utilized a large population-based sample to investigate the association between physician-diagnosed migraine and mental disorders. METHOD: Data were from the German Health Survey conducted between 1997 and 1999 (N=4181, response rate 61.4%, age 18-65 years). Lifetime and 12-month history of migraines were assessed by self-report and by a physician. Past 12-month DSM-IV mental disorders were assessed using the Composite International Diagnostic Interview. RESULTS: After adjusting for sociodemographic factors, past-year migraine was significantly and positively associated with depression, dysthymia, bipolar disorder, panic attacks, panic disorder, agoraphobia and simple phobia [adjusted odds ratios (AOR) ranging from 1.74 to 3.21]. After additionally adjusting for other mental disorders, any anxiety disorder (AOR=1.82) and any mood disorder (AOR=1.61) remained significantly associated with past-year migraine. CONCLUSION: Although causal inferences cannot be made due to the cross-sectional nature of the data, the present study adds to a growing body of literature that suggests a strong association between migraines and mood and anxiety disorders. PMID- 19134505 TI - Psychological reactions to amputation in a sample of Nigerian amputees. AB - OBJECTIVE: The study compares psychological symptoms between amputees and other orthopedic patients. METHOD: Forty-two consecutive amputees were interviewed between 7 and 28 days after amputation, and an equal number of other orthopedic patients matched for age, sex, marital status, and occupation were used as controls. Each respondent completed a sociodemographic questionnaire, while clinical variables were obtained from the case notes. Respondents also completed the General Health Questionnaire, State Trait Anxiety Inventory, and the Zung Self-Rating Depression Inventory. RESULTS: The mean age of amputees in this study was 42.33 years (S.D.=15.89 years), and the average weekly income is #3500.00 ($29.00). Anxiety and depressive symptoms were high among amputees (64.3% and 59.5%, respectively) compared to other orthopedic patients (14.3% and 12.0%, respectively). Correlation analysis showed that there was significant correlation between anxiety and age (negative), marital status, and level of education, while depressive symptoms significantly correlated significantly with age (negative) and marital status. CONCLUSION: Psychological symptoms are high in this sample of amputees, indicating the importance of social and emotional support for these patients. PMID- 19134506 TI - Prevalence and clinical profile of delirium: a study from a tertiary-care hospital in north India. AB - BACKGROUND: There is scarcity of data from the Indian subcontinent in terms of rates of psychiatric referrals and prevalence of delirium in the medico-surgical setting. AIM: The present research aimed to study the rates of psychiatric referrals, clinical profile and treatment of delirium in a tertiary-care hospital. METHODS: The referral register in the department of psychiatry (which records all referrals to the Consultation-Liaison Psychiatry Team) was used to obtain data of all referred patients who had been diagnosed to have delirium while they were admitted in the Nehru Hospital, Postgraduate Institute of Medical Education and Research, Chandigarh, during the period 2000-2005. RESULTS: Over 6 years, 3092 patients/referrals were received from different wards and included 1050 who were diagnosed as having delirium. The psychiatric referral rate ranged from 0.92% to 1.56% (mean=1.3%) per year; delirium formed the largest diagnostic category (30.77% to 38.95% of all referred cases). The prevalence of delirium in all inpatients ranged from 0.28% to 0.53% (mean=0.44%), with prevalence being higher in the elderly. In 80% of the cases, the referral was for abnormal behavior or patient's noncooperation for treatment. Most of the cases improved with treatment. CONCLUSIONS: Delirium forms the largest diagnostic category in psychiatry referrals and improves with treatment. PMID- 19134507 TI - Care and management of cardiovascular risk factors among individuals with schizophrenia and type 2 diabetes who smoke. AB - OBJECTIVE: We sought to determine whether individuals with schizophrenia and type 2 diabetes who smoke were being monitored and treated for modifiable risk factors associated with cardiovascular disease. METHODS: Cross-sectional analysis of 100 patients with schizophrenia and 99 without serious mental illness (SMI), with a diagnosis of type 2 diabetes. RESULTS: Individuals with schizophrenia were nearly twice as likely to be smokers compared to those without SMI (62% vs. 34%). Among smokers, those with schizophrenia were significantly less likely to receive blood pressure exams, lipid profiles or treatment with ACE inhibitors or statins compared to those without SMI. Both groups were equally likely to receive smoking cessation counseling. CONCLUSIONS: Smokers with type 2 diabetes and schizophrenia are significantly less likely to receive services and treatments known to improve cardiovascular outcomes. Efforts to increase awareness and improve delivery of services to this vulnerable group of patients are warranted. PMID- 19134508 TI - Health care utilisation and quality of life in individuals with diabetes and comorbid mental disorders. AB - OBJECTIVE: To investigate the association of comorbid mental disorders with health care utilisation and quality of life (QoL) in persons with diabetes (PWD). METHOD: Data were drawn from the German National Health Interview and Examination Survey (GNHIES). Mental disorders in PWD (n=146) were assessed by means of a standardised clinical interview for mental disorders (M-CIDI). Health care utilisation was assessed by using self-report questionnaires and QoL by using the SF-36. RESULTS: Controlling for age and sex, mental comorbidity was significantly associated with reduced QoL scores on all SF-36 subscales, except for "bodily pain". The frequency of physician visits was (insignificantly) higher in PWD with mental disorders compared to PWD without (20.7 vs. 14.4). Among PWD with mental disorders (n=40), only seven individuals reported mental health specialist visits. Mental comorbidity was not associated with hospitalisation days, disability days or utilisation of diabetes-related preventive services. CONCLUSIONS: Comorbid mental disorders were found to be associated with lowered psychosocial and physical aspects of QoL in a nationally representative sample of PWD. Associations with health care utilisation were less consistent. The low rate of mental health specialist visits in PWD with comorbid mental disorders may indicate an under-use of appropriate health care services. PMID- 19134509 TI - Collaborative care for adolescent depression: a pilot study. AB - OBJECTIVE: The main objectives of this study were to explore the preliminary outcomes and assess the feasibility and acceptability of a collaborative care intervention designed to improve treatment and outcomes of depression among youth seen in primary care settings. METHODS: We conducted a pilot intervention study at three clinics in a university affiliated primary care clinic network. The intervention model was designed to support the provision of depression treatment by primary care providers using methods adapted from the IMPACT study developed for the improvement of depression among older adults. Specific components include the provision of regular case management by a nurse depression care manager (DCM), enhanced patient and parent education about depression and its treatment, encouragement of patient self-management with a choice of starting medications or therapy or both, and oversight of the DCM by a mental health specialist. Study participants were assessed regularly by the DCM for 6 months and completed written self-report assessments at baseline, 3, and 6 months after starting the intervention. RESULTS: 40 youth (12-18 years) with major and minor depression enrolled in the intervention. Study participants were predominantly female (90%). The baseline Patient Health Questionnaire (PHQ-9) score was 14.2 (SD=4.5). Patients were similarly divided among initiating medications (n=12), therapy (n=15), or combination therapy (n=8). Five patients withdrew prior to initiating treatment. The mean number of in person and telephone contacts with the DCM was 9 (range=5 to 17). Eighty-seven percent of youth completed the 6-month intervention. At 6 month follow-up, 74% of youth had a 50% or more reduction in depressive symptoms as measured by the PHQ-9. Parents, youth and physicians indicated high levels of satisfaction with the intervention on written surveys and in qualitative exit interviews. CONCLUSION: The collaborative care model is feasible and highly acceptable to adolescents and parents as demonstrated both by self-report and by engagement in the intervention. It is also associated with improved depressive outcomes at similar levels to adult interventions. Future studies should evaluate these models in a randomized controlled trial. PMID- 19134510 TI - Receiving treatment for common mental disorders. AB - OBJECTIVE: Anxiety and depressive disorders are widely prevalent, but patients are only treated in a minority of cases. In this study, the explanation of receiving mental health treatment is sought in predisposing and enabling characteristics and indicators for objective and self-perceived need. METHODS: Cross-sectional analysis of data collected in the Netherlands Study of Depression and Anxiety (NESDA) among 743 persons with an anxiety and/or depression diagnosis as assessed by the CIDI. Receipt of mental health treatment was assessed in the face-to-face interview, as well as indicators of predisposing and enabling factors and variables evaluating need for care. RESULTS: Of the total sample, 57% received treatment in the past 6 months in the general practice setting (50%) or the mental health care setting (14%). Younger patients, patients who evaluated their providers better on communicative abilities and patients who perceived mental health problems themselves had greater odds of having professional mental health contacts in the primary care setting. Confidence in professional help and higher severity of mental problems were associated with greater odds of having specialized mental health care. CONCLUSION: Receiving help for common mental disorders depends not only on the objective need of the patient but also at least as much on the patients' own recognition that their problems have a mental health origin. Furthermore, in primary care especially, the patients' judgment of their providers' affective abilities may be decisive for being treated. For receiving specialized care, patients are also directed by their confidence in professional help. PMID- 19134511 TI - Validation of the Leverton Questionnaire as a screening tool for postnatal depression in Hungary. AB - OBJECTIVE: To assess the validity of the 24-item Leverton Questionnaire (LQ) in screening for postnatal depression (PND). METHOD: A two-phase, cross-sectional study was designed. Between January and October 2006, a sample of 1552 women attending a routine postnatal check-up at 6 weeks postpartum completed the LQ in southeast Hungary. On the basis of the LQ total score, the participants were stratified and randomly selected within each stratum for clinical evaluation (Structured Clinical Interview for DSM-IV). Receiver operating characteristic (ROC) analyses were used to examine the sensitivity and specificity of the LQ to detect PND. RESULTS: The best cut-off on the Hungarian version of the LQ for PND was 11/12, with a sensitivity of 88.0%, and a specificity of 94.4%, and a positive predictive value of 53.1%. Internal consistency was satisfactory (Cronbach alpha coefficients > or = 0.753). The sensitivity of the modified Beck Depression Inventory (BDI) in detecting PND was 86.2% and the specificity 90.4%. Although the BDI performed slightly better than the LQ in distinguishing between minor and major depression, both psychometric scales showed satisfactory screening performance. CONCLUSIONS: Our data confirm the validity of the Hungarian version of the LQ reliably to identify PND. We propose a cut-off of 11/12 for screening purposes for PND, the range of 11-14 for detecting minor depression and regarding a total score of 15 points or above as indicative of major depression. PMID- 19134512 TI - Cause-specific excess mortality in suicidal patients: gender differences in mortality patterns. AB - OBJECTIVE: Few studies have investigated gender-specific excess mortality in suicidal patients. The objective of this study was to examine the risk of dying from all causes of death in male and female suicidal patients. METHOD: This study included 4140 patients who received psychiatric treatment in two Austrian hospitals after non-fatal suicidal behavior between 1989 and 2001. Mortality was prospectively assessed for a maximum follow-up period of 17 years (1989-2005). Gender-specific standardized mortality ratios were calculated for all causes of death. Rate ratios were used to assess gender differences. RESULTS: Males but not females had an elevated risk to die from "natural" causes of death. Mortality from "unnatural" causes of death, particularly suicide, was elevated in both genders. The female suicide risk markedly exceeded the risk for males. In both genders, mortality caused by substance use disorders was considerably elevated. CONCLUSION: Significant gender differences in the excess mortality of suicidal patients underline the need for gender-specific research in psychiatry. PMID- 19134513 TI - Reliability and validity of the Japanese version of the Peritraumatic Distress Inventory. AB - OBJECTIVE: To assess the reliability and validity of the Japanese version of the Peritraumatic Distress Inventory (PDI). METHOD: One hundred thirty-five participants with physical injury resulting from motor vehicle accidents were consecutively recruited in this cross-sectional study, from Aug. 18, 2005, to Jan. 8, 2008. A subsample (n=71) were retested on the PDI an average of 96.4 days after initial measure completion. RESULTS: Correlational analyses revealed an overall Cronbach's alpha coefficient of 0.83. The item-total correlations for the 13 items ranged from 0.29 to 0.75. The test-retest correlation coefficient was 0.61. The PDI was significantly correlated with the external validators such as peritraumatic dissociation as measured by the Peritraumatic Dissociative Experiences Questionnaire (PDEQ); the intrusion, avoidance and hyperarousal scores of the Impact of Events Scale-Revised (IES-R); and the depression and anxiety subscales of the Hospital Anxiety Depression Scale (HADS) (P<.01). CONCLUSION: The present study indicated that the Japanese version of the PDI has a high degree of internal consistency, acceptable reliability and a high degree of concurrent validity with measures of peritraumatic dissociation and posttraumatic symptoms. The Japanese version of the PDI can be used as a validated instrument in future research. PMID- 19134514 TI - Wards features associated with high rates of medication refusal by patients: a large multi-centred survey. AB - OBJECTIVE: This article aimed to explore the relationship of medication-related conflict [refusal of regular medication, refusal of pro re nata (prn) medication, demanding prn medication] to other conflict behaviours, the use of containment methods, service environment, physical environment, patient routines, staff demographics and staff group variables. METHOD: The Patient-staff Conflict Checklist (PCC-SR), an end-of-shift report completed by nurses on the frequency of conflict and containment events, was collected for a 6-month period on 136 acute mental health wards in 67 hospitals within 26 NHS Trusts in England, in 2004-2005. Multilevel modelling was used to assess associations with medication related conflict rates. RESULTS: The mean daily rate (at ward level, standardised to 20 beds) of incidents of regular medication refusal was 0.89 (S.D. 0.52), prn medication refusal 0.30 (S.D. 0.19) and demanding prn medication 1.09 (S.D. 0.63). The frequency of these events was found to be associated with passive resistant patient behaviours, higher levels of containment (specifically locking the main ward door, the use of special observation, and time out) and unstable staffing profiles. CONCLUSION: It may be possible to achieve greater medication concordance amongst patients in acute mental health wards through a more consensual approach to care. Paradoxically, fewer restrictions may promote better treatment acceptance and safer outcomes. Consistent nurse staffing and therefore better staff-patient relationships are also likely to improve cooperation and outcomes. PMID- 19134515 TI - Catatonia and systemic lupus erythematosus: a clinical study of three cases. AB - Catatonia may be encountered in psychiatric disorders, but also in general medical conditions. Cases of catatonia associated with systemic lupus erythematosus (SLE) are rare. Several articles have described this symptomatic association, as well as its management, using electroconvulsive therapy, plasma exchange or benzodiazepines. We report three cases here of patients who presented with catatonia during a lupus relapse, in whom treatment with lorazepam improved the catatonic symptomatology, thus allowing the associated condition to be treated. We touch on several points about the diagnosis, etiology and treatment of catatonia, when it is associated with SLE. PMID- 19134516 TI - Clozapine-induced severe mixed hyperlipidemia: a case report. AB - OBJECTIVE: To highlight the association between the use of clozapine and the early development of severe mixed hyperlipidemia, a condition that substantially increases the risk of cardiovascular events and other medical complications. METHOD: Clinical information and data from investigations in a single patient were collated and reviewed in light of the literature. RESULTS: A 47-year-old man with a background history of schizophrenia taking clozapine presented with multiple episodes of severe elevation of serum triglycerides and cholesterol with subsequent development of diabetes mellitus. His metabolic parameters normalized following discontinuation of clozapine. CONCLUSION: Lipid and metabolic profiles should be closely monitored in patients receiving clozapine in order to facilitate early detection and intervention to prevent further health complications. PMID- 19134517 TI - Risperidone-induced thrombocytopenia: a case report. PMID- 19134518 TI - Psychiatric treatment of patients predisposed to priapism induced by quetapine, trazadone and risperidone: a case report. PMID- 19134519 TI - Therapeutic challenges in the era of antibiotic resistance. AB - Multidrug microbial resistance poses major challenges to the management of infection, particularly with the paucity of new drugs with activity against these bacteria. Since the turn of this century a few new antibiotics have been licensed, including linezolid, daptomycin and tigecycline. This supplement reports data presented at the 13th International Congress of Infectious Diseases held in Kuala Lumpur in June 2008. Dr R. Isturiz reviews the data on global resistance trends and the potential impact on empirical therapy; Dr J.-H. Song reviews new agents on the antimicrobial horizon; and the final paper in the supplement, by Dr L.R. Peterson, reviews the role of tigecycline in the management of complicated intra-abdominal and skin and soft tissue infections. PMID- 19134520 TI - Global resistance trends and the potential impact on empirical therapy. AB - Globally, antibiotic resistance continues to be an increasing problem. The emergence of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin resistant enterococci (VRE), and extended spectrum beta-lactamase-producing Enterobacteriaceae has seriously reduced the number of empirical agents suitable for selected indications. Increasing Gram-negative resistance has also negatively impacted the physician's alternatives when choosing adequate initial therapy; increased reliance on the carbapenem class as empirical agents has led to the emergence of multidrug resistance in non-enteric Gram-negative bacilli. There is a critical need for new, broad-spectrum alternatives to the currently available broad-spectrum agents. PMID- 19134521 TI - What's new on the antimicrobial horizon? AB - The antimicrobial era is threatened by high levels of antibiotic resistance, the limited number and disparate availability of effective antibiotics against diverse bacterial species, and reduced involvement by the pharmaceutical industry in the development of new anti-infectives. For the treatment of resistant Gram positive coccal infections, particularly methicillin-resistant staphylococcal infections, vancomycin has long been the mainstay antimicrobial agent due to its safety, durability against resistance, and lack of other approved alternatives. However, the efficacy and safety of vancomycin for the treatment of many serious infections has been called into question. Promising results from clinical trials suggest that five new antimicrobials could offer safe and effective alternatives to vancomycin. With regard to resistant Gram-negative infections, new carbapenems and some other options will be available. This paper reviews the safety and efficacy of these new antimicrobial agents against resistant bacterial pathogens. PMID- 19134522 TI - A review of tigecycline--the first glycylcycline. AB - The dawn of a troubling post-antibiotic era likely is on the horizon, fuelled by a rise in bacterial resistance to existing antibiotic therapy alongside a waning pipeline of novel antibacterial agents. Tigecycline, a new glycylcycline with an expanded broad spectrum of in vitro activity, was recently approved for the treatment of complicated skin and soft tissue infections (cSSTIs) and complicated intra-abdominal infections (cIAIs). This review will examine how tigecycline evades the common mechanisms of antibiotic resistance, the metabolism and pharmacokinetics of tigecycline, and its spectrum of in vitro activity. The results of randomized clinical trials for the treatment of cSSTIs and cIAIs with tigecycline are also described, as is the patient safety and tolerability observed during these studies. Tigecycline monotherapy has been shown to be as effective as its comparators and, against a backdrop of rising bacterial resistance, the role for tigecycline in monotherapy of infections from Gram positive, Gram-negative and anaerobic bacteria is a meaningful development. PMID- 19134523 TI - Lung transplantation for lymphangioleiomyomatosis: the European experience. AB - BACKGROUND: Lung transplantation has been accepted widely as therapy for end stage pulmonary lymphangioleiomyomatosis (LAM); however, single-center and national experience is limited due to the rarity of LAM. METHODS: We report the recent European experience of lung transplantation for LAM. A self-administered questionnaire was distributed to 30 European lung transplant centers to evaluate patients who underwent primary lung transplantation for LAM (1997 to 2007). RESULTS: Seventy percent of centers responded to the questionnaire. A total of 61 lung transplants were undertaken in women only, with mean age at transplant 41.3 years (SD 5.1). Centers performed a median of 2 (0 to 9) transplant operations. Severe pleural adhesions were the most common intra-operative complication. Early deaths (N = 6) were due to primary graft or multiple-organ failure or sepsis. Twelve recipients were diagnosed with bronchiolitis obliterans syndrome at a median of 20 months (range 10 to 86 months) post-transplant. LAM-related complications included renal angiomyolipoma and pneumothorax in the native lung. Recurrence of LAM occurred in 4 recipients. As of December 2007, actuarial Kaplan Meier survival was 79% at 1 year and 73% at 3 years post-transplant. CONCLUSIONS: Post-transplant outcome for pulmonary LAM in the recent era appears to have improved compared with the previous era. LAM-related complications remain common, but recurrence of LAM in the allograft is rare. PMID- 19134524 TI - Lung transplant outcomes in white and African American recipients: special focus on acute and chronic rejection. AB - BACKGROUND: The effects of lung transplant recipient race on post-transplant outcomes have not been adequately evaluated. This cohort study seeks to determine the characteristics of African American lung transplant recipients and the effects of African American race on post-transplant outcomes, particularly acute and chronic rejection, compared with white recipients, at a single center. METHODS: There were 485 first-time lung transplantations (431 white, 47 African American, 5 Hispanic and 2 Asian recipients) performed at the University of Pennsylvania between 1991 and 2006. All white and African American recipients were compared based on pre-transplant diagnoses and post-transplant survival. The cohort from 1998 to 2006 (239 white and 25 African American recipients) was also compared based on acute rejection score (ARS) and development of bronchiolitis obliterans syndrome (BOS). RESULTS: Chronic obstructive pulmonary disease was the most common diagnosis leading to lung transplantation in both groups, but sarcoidosis was a much more common indication in African American recipients (white, 1%; African American, 28%; p < 0.001). Survival was similar in the two groups (white vs African American groups: 1 month, 90.0% vs 87.2%; 1 year, 74.9% vs 74.5%; 5 years, 52.3% vs 50.5%, respectively; p = 0.84). Freedom from BOS at 3 years (white, 60.3%; African American, 62.8%; p = 0.30) and ARS per biopsy (white, 0.83 +/- 0.82; African American, 0.63 +/- 0.77; p = 0.31) were similar in both groups. CONCLUSIONS: White and African American patients seek lung transplantation for different diseases, but post-transplant outcomes were found to be similar. Larger, multi-center studies are needed to confirm these results. PMID- 19134525 TI - Single-institution study evaluating the utility of surveillance bronchoscopy after lung transplantation. AB - BACKGROUND: Many lung transplant physicians advocate surveillance bronchoscopy with transbronchial lung biopsy and bronchoalveolar lavage (TBB/BAL) to monitor lung recipients despite limited evidence this strategy improves outcomes. This report compares rates of infection (INF), acute rejection (AR), bronchiolitis obliterans syndrome (BOS) and survival in lung allograft recipients managed with surveillance TBB/BAL (SB) versus those with clinically indicated TBB/BAL (CIB). METHODS: We reviewed 47 consecutive recipients transplanted between March 2002 and August 2005. Of these recipients, 24 consented to a multi-center trial requiring SB and 23 were managed by our usual practice of CIB. Rates of freedom from INF, AR, BOS and survival were compared. BOS and AR were diagnosed according to published guidelines from the International Society for Heart and Lung Transplantation. RESULTS: A total of 240 TBB/BALs were performed. CIB and SB groups underwent 84 (3.7 +/- 3.4/patient) and 156 (6.5 +/- 2.0/patient) TBB/BALs, respectively. In the SB group, 54 (2.2 +/- 1.6/patient) TBB/BALs were true surveillance procedures, whereas 102 (4.2 +/- 2.3/patient) were clinically indicated. No AR episode requiring treatment was detected by true surveillance. Freedom from respiratory INF, AR, BOS and survival in the SB and CIB groups showed no significant differences. Five patients in the CIB group remained stable without requiring TBB/BAL. In the SB group, 4 previously asymptomatic patients developed pneumonia within 2 weeks of surveillance TBB/BAL. CONCLUSIONS: With no obvious advantage identified, surveillance bronchoscopy may pose a risk to stable lung transplant recipients. A multi-center, controlled trial is required to validate the utility and safety of surveillance bronchoscopy in lung transplantation. PMID- 19134526 TI - Increased pulse wave velocity and blood pressure in children who have undergone cardiac transplantation. AB - BACKGROUND: Arterial hypertension and premature coronary artery disease are poorly understood complications of cardiac transplantation in children. Arterial stiffness is associated with cardiovascular risk in adults. Pulse wave velocity (PWV) may be used as a surrogate for arterial rigidity. In this study we investigate PWV in children after cardiac transplantation. METHODS: Sitting blood pressure was measured in 22 children (>6 months after transplantation) and 95 controls and PWV was measured using the SphygmoCor device by high-fidelity applanation tonometry at the carotid, radial and femoral arteries. RESULTS: The transplant group was significantly older than the control group (13.4 years vs 11.1 years; p = 0.006), but there was no significant height or weight difference. The diastolic (but not systolic) pressure was significantly higher in the transplant group (75 mm Hg vs 65 mm Hg; p = 0.003). Aortic (carotid/femoral) PWV was significantly associated with age, height, weight (in the control group only) and systolic blood pressure according to univariate analysis, whereas brachial (carotid/radial) PWV was unrelated to these. According to multivariate analysis, height accounted best for all relationships with aortic PWV, and age and weight for brachial PWV. Using multivariate analysis, PWV was significantly higher in the cardiac transplant group for brachial (7.6 m/s vs 6.6 m/s; p < 0.01) and aortic (5.3 m/s vs 4.7 m/s; p < 0.001) measurements. The relation between length of time since transplantation and aortic PWV was statistically significant (p < 0.01). CONCLUSIONS: Arterial rigidity is increased in children after cardiac transplantation. An improved understanding of blood pressure and arterial stiffness may help inform the choice of blood pressure medication in these patients. PMID- 19134527 TI - Assessment of the Cylex ImmuKnow cell function assay in pediatric heart transplant patients. AB - BACKGROUND: The Cylex ImmuKnow (Cylex, Columbia, MD) cell function assay (CICFA) is a commercially available test of immune response that purportedly identifies solid organ transplant patients at risk for either acute rejection (AR) or infection. Data on the utility of this test in pediatric heart transplant patients are very limited. This study tested the hypothesis that CICFA is a clinically useful test in this transplant population. METHODS: All children undergoing heart transplantation at the study center (1989-2006) for whom CICFA levels were obtained were reviewed. The association of CICFA levels with episodes of AR and significant infections was determined. RESULTS: Among 83 patients (34 girls, 41%), 367 CICFA levels were obtained (median, 4.0; interquartile range [IQR], 2.0-6.0 per patient). There were 26 episodes of AR in 17 patients (20%) and 38 infections in 34 patients (41%). CICFA levels were similar among patients with AR at the time of the CICFA measurement (median, 325 [IQR, 163-480] adenosine triphosphate [ATP] ng/ml) vs patients without AR (median, 330 [IQR, 227 441] ATP ng/ml; p = 0.36). CICFA levels were similar among patients with infections within 1 month of CICFA measurement (median, 295 [IQR, 216-366] ATP ng/ml) and those without infections (median, 330 [IQR, 226-453] ATP ng/ml; p = 0.24). CONCLUSIONS: The CICFA is not predictive of AR or significant infections in pediatric heart transplant patients. On the basis of the available evidence, this assay cannot be recommended as part of the routine management of pediatric heart transplant patients. PMID- 19134528 TI - Cost utility evaluation of extracorporeal membrane oxygenation as a bridge to transplant for children with end-stage heart failure due to dilated cardiomyopathy. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) and cardiac transplantation are recognized to be expensive. METHODS: We performed a cost utility evaluation with a decision model approach, including 75 children with dilated cardiomyopathy. A cohort of patients with end stage heart failure who were offered ECMO bridging was compared with a similar cohort offered only conventional intensive care. Outcome was measured in cost per quality adjusted life year (QALY). RESULTS: Median follow-up was 4.39 years (interquartile range, 1.83-5.74 years), during which 50 children underwent transplantation, 16 had a period of recovery, and 25 died. ECMO bridging was highly effective (hazard ratio, 0.181; 95% confidence interval, 0.067-0.489; p = 0.001) but exceeded conventional criteria for cost-effectiveness. The reference incremental cost effectiveness ratio (ICER) was pound65,645 per QALY and pound54,284 per life-year gained. Average life expectancy rose from 6.78 to 9.79 years and costs from pound146,398 to pound309,599 per patient with ECMO bridging. The ICER was sensitive to ECMO cost, the long-term transplant survival rate, and quality of life in transplant recipients. CONCLUSIONS: ECMO bridging is effective but expensive. The eligible target population is small, nationally, positively influencing affordability. We strongly support our national policy of mechanical bridge to transplant for suitable children in end stage heart failure. Cost effectiveness could be optimized by: 1) increased availability of organ donors, 2) reduction in mechanical support costs possibly by alternate devices and 3) inclusion of patients most likely to benefit. PMID- 19134529 TI - Intraoperative evaluation of the HeartMate II flow estimator. AB - BACKGROUND: Direct measurement of blood flow output has been incorporated into ventricular assist devices (VADs), but long-term reliability of the additional device components has raised concerns regarding sensor drift and failure. As an alternative approach, the HeartMate II axial VAD (Thoratec Corp, Pleasanton, CA) estimates device flow output from power consumption and rotational speed of the device motor. This study evaluated the accuracy of HeartMate II flow estimation at the time of implantation. METHODS: In 20 patients, intraoperative blood flow measurement of the HeartMate II flow estimator was compared with flow values obtained with an ultrasonic flow probe placed around the device outflow graft. Estimated and measured VAD flow data were simultaneously recorded and digitally stored while the device motor speed varied from 7,800 to 11,000 rpm and while achieving device flow outputs of 2 to 7 liters/min. Estimated and measured flows were compared using linear regression analyses and root mean square error. RESULTS: HeartMate II flow estimation (FE) demonstrated a linear correlation with ultrasonic flow probe (FP) measurements: FE = 0.74 FP + 0.99 (R(2) = 0.56, p = 0.0001). A root mean square error of 0.8 liters/min was observed between flow estimation and direct flow measurement and suggests a 15% to 20% difference at flows of 4 of 6 liters/min. CONCLUSIONS: These results suggest that HeartMate II flow estimation may be used to provide directional information for trend purposes rather than absolute values of device blood flow output. Patient management should include but not be limited to this information. PMID- 19134530 TI - Predictors of death and transplant in patients with a mechanical circulatory support device: a multi-institutional study. AB - BACKGROUND: INTERMACS is a registry of FDA-approved durable mechanical circulatory support (MCS) devices used for the strategies of destination therapy (DT) and bridge to transplantation (BTT) or recovery. This study identifies predictors for death and transplantation based on initial results from INTERMACS. METHODS: From June 23, 2006 to December 31, 2007, 420 patients from 75 institutions were prospectively entered into the INTERMACS database in which pre implant data, indication for MCS device use, adverse events, demographics, hemodynamics, laboratory values and outcomes were recorded. Using competing outcomes methodology, risk factors were identified for the events of death and transplantation. RESULTS: The devices included 314 left ventricular assist devices (LVADs), 5 right VADs (RVADs), 77 biventricular VADs (biVADs) and 24 total artificial hearts (TAHs) for a total of 497 pumps in 420 patients. Among the BTT patients at 6 months, 33% were alive with a device in place, 42% were transplanted, 22% had died, and 3% were explanted for recovery. Among the DT patients at 6 months, 68% were alive with a device in place, 5% were transplanted, 25% had died, and 2% were explanted for recovery. The risk factors identified for death across all patient groups include older age (relative risk [RR] = 1.41, p < 0.001), ascites (RR = 2.04, p = 0.003), increased bilirubin (RR = 1.49, p < 0.05) and INTERMACS Level 1 (cardiogenic shock) (RR = 1.59, p = 0.02). The most common causes of death were central nervous system (CNS) event (18.3% of deaths), multiple-organ failure (16.4%) and cardiac cause (right ventricular failure and arrhythmias, 15.4%). CONCLUSIONS: Cardiogenic shock, advanced age and severe right heart failure manifested as ascites or increased bilirubin are risk factors for death after MCS therapy. BTT patients who require biVAD support have a transplant rate similar to that of LVAD-only patients, but their mortality at 6 and 12 months exceeds that of LVAD-only patients. Consideration should be given to MCS referral before the sequelae of right ventricular failure dominate the advanced heart failure syndrome. PMID- 19134531 TI - A clinical correlation study of severity of antibody-mediated rejection and cardiovascular mortality in heart transplantation. AB - BACKGROUND: The current International Society for Heart and Lung Transplantation (ISHLT) diagnostic criteria for antibody-mediated rejection (AMR) designate AMR as either absent (AMR 0) or present (AMR 1), without grading its severity. Yet, the extent of histologic and immunofluorescence (IF) findings of AMR varies across endomyocardial biopsies (EMBs). In this study, we hypothesized that the severity of AMR, as assessed on EMBs, correlates with cardiovascular mortality in heart transplant recipients. METHODS: All EMBs from 1985 to 2005 were evaluated. Biopsy specimens were uniformly studied by light microscopy and IF early post transplant. A comprehensive vascular score (V1: no AMR, to V5: severe AMR) was prospectively assigned to each EMB, based on severity of both histologic and IF findings. Univariate Cox proportional hazards regressions were performed using indicators of vascular scores alone, combined, and cumulatively. RESULTS: Nine hundred six patients were transplanted and included in the study. Mean age was 46.6 +/- 15.5 years and 82% were male. A total of 26,236 EMBs comprised the study data. As expected, histologic and immunopathologic findings of AMR varied in severity. An incremental risk of cardiovascular mortality was found with more severe AMR whether vascular scores were analyzed individually (p = 0.001), in combination (p = 0.01) or cumulatively (p = 0.006). CONCLUSIONS: The severity of AMR on EMBs correlates with an incremental cardiovascular mortality risk after heart transplantation, suggesting that AMR should be viewed as a spectrum rather than just as present or absent. Supplementing the ISHLT AMR diagnostic guidelines with a consensus severity scale is warranted. PMID- 19134532 TI - Tacrolimus versus cyclosporine microemulsion for heart transplant recipients: a meta-analysis. AB - BACKGROUND: Tacrolimus and cyclosporine microemulsion are the 2 major immunosuppressants for heart transplantation. Several studies have compared these 2 drugs, but the outcomes were not consistent. This meta-analysis of randomized controlled trials compared the beneficial and harmful effects of tacrolimus and microemulsion cyclosporine for heart transplant recipients. METHODS: Electronic databases and manual bibliography searches were conducted. A meta-analysis was performed of all randomized controlled trials comparing tacrolimus with cyclosporine microemulsion as primary immunosuppression for heart transplant recipients. Data for mortality, acute rejection, withdrawals, and adverse events were extracted. The combined results of the data of the randomized controlled trials were summarized as relative risk with 95% confidence intervals. RESULTS: The study assessed 7 randomized controlled trials including 885 patients. There was no difference in mortality at 1 year between recipients treated with tacrolimus and cyclosporine microemulsion. Tacrolimus-treated patients had less acute rejection risk at 6 months and 1 year. Fewer patients stopped tacrolimus than cyclosporine microemulsion. The rate of new-onset diabetes mellitus requiring insulin treatment was higher with tacrolimus. More post-transplantation hypertension occurred with cyclosporine microemulsion. The groups had comparable incidences of malignancy and renal failure needing dialysis. CONCLUSIONS: The use of tacrolimus as primary immunosuppressant for heart transplant recipients results in comparable survival and a significant reduction in acute rejection compared with cyclosporine microemulsion. PMID- 19134533 TI - Efficacy of oral ribavirin in lung transplant patients with respiratory syncytial virus lower respiratory tract infection. AB - BACKGROUND: Respiratory syncytial virus (RSV) can cause severe lower respiratory tract infection (LRI) and is a risk factor for the development of bronchiolitis obliterans syndrome (BOS) after lung transplantation (LTx). Currently, the most widely used therapy for RSV is inhaled ribavirin. However, this therapy is costly and cumbersome. We investigated the utility of using oral ribavirin for the treatment of RSV infection after LTx. METHODS: RSV was identified in nasopharyngeal swabs (NPS) or bronchoalveolar lavage (BAL) using direct fluorescent antibody (DFA) in 5 symptomatic LTx patients diagnosed with LRI. Data were collected from December 2005 and August 2007 and included: age; gender; type of LTx; underlying disease; date of RSV; pulmonary function prior to, during and up to 565 days post-RSV infection; need for mechanical ventilation; concurrent infections; and radiographic features. Patients received oral ribavirin for 10 days with solumedrol (10 to 15 mg/kg/day intravenously) for 3 days, until repeat NPS were negative. RESULTS: Five patients had their RSV-LRI diagnosis made at a median of 300 days post-LTx. Mean forced expiratory volume in 1 second (FEV(1)) fell 21% (p < 0.012) during infection. After treatment, FEV(1) returned to baseline and was maintained at follow-up of 565 days. There were no complications and no deaths with oral therapy. A 10-day course of oral ribavirin cost $700 compared with $14,000 for nebulized ribavirin at 6 g/day. CONCLUSIONS: Treatment of RSV after LTx with oral ribavirin and corticosteroids is well tolerated, effective and less costly than inhaled ribavirin. Further studies are needed to directly compare the long-term efficacy of oral vs nebulized therapy for RSV. PMID- 19134534 TI - Cell-mediated immunomodulation of chemokine receptor 7-expressing porcine sertoli cells in murine heterotopic heart transplantation. AB - BACKGROUND: Sertoli cells (SC) have immunomodulative properties, and chemokine receptor 7 (CCR7) can optimize the systemic immunomodulatory effect by guiding SC from the periphery to the secondary lymphoid organs. METHODS: The effect of immortalized neonatal porcine SC (NPSCi) was evaluated by analysis of cytokine levels. Hyporesponsiveness to donor cells was determined by MLC and analysis of splenocyte phenotypes using a murine allogeneic skin graft model. The effect of CCR7-expressing NPSCi (NPSCi-CCR7) combined with cobra venom factor (CVF) was evaluated using a heterotopically transplanted murine allogeneic heart model. RESULTS: Expression of immune cytokines was markedly modulated by NPSCi. The lymphocyte proliferation and splenocyte phenotypes were significantly suppressed by NPSCi-CCR7. Although pre-transplantation of NPSCi or NPSCi-CCR7 did not prolong graft survival of allogeneic cardiac grafts, CVF treatment facilitated pre-transplantation of NPSCi-CCR7 to prolong survival of allogeneic cardiac grafts (25.5 +/- 7.05 vs 9.5 +/- 0.58 days, p < 0.01). CONCLUSIONS: NPSCi may be used as a powerful immunomodulatory tool, and our strategy to traffic NPSCi to lymphoid organs using CCR7 optimizes the systemic immunomodulatory effect in vivo. With the help of initial immunosuppression for humoral mechanisms using CVF, the host immune response against allogeneic cardiac grafts can be effectively ameliorated by immunomodulation of the cellular mechanism with NPSCi CCR7. PMID- 19134535 TI - Nebulized phosphodiesterase III inhibitor during warm ischemia attenuates pulmonary ischemia-reperfusion injury. AB - BACKGROUND: The control of warm ischemia-reperfusion injury is crucial in managing donors after cardiac death for lung transplantation. We focused on transalveolar administration as a drug-delivery route for such donors. Milrinone is a phosphodiesterase 3 inhibitor that inhibits the breakdown of cyclic adenosine monophosphate and selectively relaxes smooth muscle. We hypothesized that nebulized milrinone would mitigate warm ischemia-reperfusion injury of lung. METHODS: This study was conducted with an isolated rat lung perfusion model. Lungs were excised, exposed to 55-minute ischemia at 37 degrees C, and reperfused for 60 minutes. During ischemia, nebulized milrinone (n = 6) or saline (n = 6) was inhaled. Lungs were continuously perfused without ischemia as a sham group (n = 6). Airway resistance, pulmonary vascular resistance, pulmonary compliance, weight gain and blood gas were measured. Adenine nucleotide levels and apoptosis were investigated in the reperfused lungs. RESULTS: Milrinone nebulization decreased post-ischemic pulmonary vascular resistance (0.98 +/- 0.05 and 1.74 +/- 0.17 cm H(2)O/ml.min at 60 minutes of reperfusion in the milrinone and control groups, respectively [p < 0.01]). It did not alter cyclic adenosine monophosphate levels, but it did elevate adenosine triphosphate levels (9.87 +/- 0.38 and 6.91 +/- 0.45 in the milrinone and control groups, respectively [p < 0.01]) and suppressed apoptosis (3.83 +/- 0.91 and 46.17 +/- 3.39 of mean apoptotic cell numbers in the milrinone and control groups, respectively [p < 0.01]). CONCLUSIONS: Milrinone nebulization decreased post-ischemic pulmonary vascular resistance, elevated adenosine triphosphate levels, and suppressed apoptosis. Nebulized milrinone has some protective effects against warm ischemia. PMID- 19134536 TI - Interleukin-6 deficiency fails to prevent chronic rejection after aortic allografts in apolipoprotein E-deficient mice. AB - BACKGROUND: Chronic vascular rejection (CVR) is characterized by an intimal thickening in the arteries of allografts due to immunoinflammatory reactions and smooth muscle cell proliferation. Interleukin 6 (IL-6) levels are increased in patients with graft rejection, however the role of IL-6 in CVR is not known. We investigated if IL-6 deficiency in the recipient could prevent CVR after an aortic allograft. METHODS: Donor aortas from wild-type DBA/2 mice were transplanted into C57BL/6 recipients, either wild-type mice or mice deficient for IL-6 (IL-6(-/-)), apolipoprotein E (ApoE(-/-)), or both (IL-6(-/-)ApoE(-/-)). Alloantibody titers were determined at Day 30, 60, or 90 after grafting. The grafts were examined for CVR lesions by morphometry and immunohistology. RESULTS: All recipient allografts displayed lesions typical for CVR. The lesions were larger in IL-6-deficient strains, and significantly so in IL-6(-/-)ApoE(-/-) recipients. Early immunoglobulin (Ig) G1 alloantibody deposits were observed in the grafts of ApoE-deficient strains and late IgG2a deposits in the grafts of IL 6-deficient strains. A rapid and sustained type 1 helper T cell (Th1; IgG2a) alloresponse in IL-6(-/-) mice, and a strong type 2 helper T cell (Th2; IgG1) response in ApoE(-/-) mice were observed. IL-6(-/-)ApoE(-/-) mice displayed the highest alloantibody titer, with a Th1 dominance. CONCLUSIONS: Unexpectedly, IL-6 deficiency in the recipient mice did not prevent CVR lesions but even aggravated them in IL-6(-/-)ApoE(-/-) recipients. This was associated with increased local and systemic alloresponses. PMID- 19134537 TI - Transplant recipients on the edge of the hypocalcemia abyss. AB - Hypovitaminosis D is a risk factor for transplant-related osteoporosis. Its contribution to severe hypocalcemia in transplant recipients is less well recognized. We present 2 cases to illustrate how risk factors specific to transplant recipients significantly increase the risk of development of severe hypocalcemia, on a background of unrecognized vitamin D deficiency. Regular surveillance of calcium homeostasis should be incorporated into routine clinical care of transplant recipients. PMID- 19134538 TI - Acute antibody-mediated rejection after lung transplantation. AB - The role of humoral immunity after lung transplantation remains unclear. In this report, we describe the pathologic findings and clinical course of a case of acute antibody-mediated rejection (AMR) after lung transplantation. After an uncomplicated early course, a 31-year-old man with cystic fibrosis developed acute graft dysfunction 1 month after bilateral lung transplantation. Lung biopsies showed acute pneumonitis with capillary injury, neutrophilic infiltration and nuclear dust. Immunostaining for C4d demonstrated endothelial cell deposition, and circulating donor-specific human leukocyte antigen (HLA) antibodies were identified. Despite severe hypoxemic respiratory failure, he responded well to a regimen consisting of methylprednisolone, plasma exchange, intravenous immunoglobulin and rituximab therapy. He completely recovered clinically although donor-specific HLA antibodies have remained detectable. The incidence of acute AMR after lung transplantation is unknown, but this case fulfills all of the consensus diagnostic criteria, and we suggest that AMR could be an under-recognized cause of acute graft dysfunction. PMID- 19134539 TI - Recovery from anthracycline cardiomyopathy after long-term support with a continuous flow left ventricular assist device. AB - We report the clinical course of a 16-year-old girl in remission from non Hodgkin's lymphoma who presented in cardiogenic shock due to a severe anthracycline cardiomyopathy. The patient was initially stabilized using central extracorporeal membrane oxygenation support, followed by conversion to a left ventricular assist device. Unexpected evidence of cardiac recovery 9 months after implant enabled device weaning during a 3-month period, culminating in successful device explantation 1 year after implant. The patient survives 18 months after explant in New York Heart Association class I, on conventional heart failure medical management and metabolic therapy. PMID- 19134540 TI - Everolimus-associated pneumonitis in 3 heart transplant recipients. AB - Although pulmonary toxicity from sirolimus is well recognized, the biochemically homologous everolimus has until recently been thought to be free from this side effect. Emerging evidence suggests an association between high-dose everolimus and symptomatic pneumonitis. Toxicity at typical immunosuppressive doses has also been described. In support of these observations, we report the occurrence of clinical pneumonitis in 3 patients in association with everolimus therapy after heart transplantation in the absence of other causative factors. Typical onset consisted of dyspnea, hypoxemia, and bilateral diffuse pulmonary infiltrates beginning between 2 weeks and 6 months after commencement of the drug. Although uncommon, everolimus may cause pneumonitis and should be considered in the differential diagnosis of pulmonary infiltrates and hypoxemia in an appropriate clinical setting. PMID- 19134541 TI - Novel approach to management of acute pulmonary failure during biventricular assist device insertion. AB - A shunt from a right ventricular assist device (BVS5000; Abiomed, Inc., Danvers, MA) with an oxygenator to a left ventricular assist device (AB5000; Abiomed) was created to circumvent the lungs in a 16-year-old boy with acute pulmonary failure during the transition from extracorporeal membrane oxygenation to biventricular support. PMID- 19134543 TI - Atorvastatin pleiotropic effects mediated via suppression of TNF-alpha in congestive heart failure patients with initial normal TNF-alpha levels. PMID- 19134542 TI - Acute constrictive pericarditis after lung transplantation for lymphangioleiomyomatosis. AB - Lymphangioleiomyomatosis (LAM) is a rare, cystic, progressive lung disease with many extrapulmonary manifestations, which may complicate allograft function after transplantation. We present a LAM patient with new dyspnea and declining spirometry without rejection, infection or recurrence one year after bilateral lung transplantation. Investigation revealed acute constrictive pericarditis, which has not been reported previously in a lung transplant patient with LAM. This represents a novel complication likely due to progression of extrapulmonary LAM and should be considered in LAM transplant patients with dyspnea. PMID- 19134544 TI - Ideal volume and perfusion pressure of cardioplegic solution during retrieval of donor heart graft. PMID- 19134545 TI - Monitoring tissue engineering using magnetic resonance imaging. AB - Assessment of tissue regeneration is essential to optimize the stages of tissue engineering (cell proliferation, tissue development and implantation). Optical and X-ray imaging have been used in tissue engineering to provide useful information, but each has limitations: for example, poor depth penetration and radiation damage. Magnetic resonance imaging (MRI) largely overcomes these restrictions, exhibits high resolution (approximately 100 microm) and can be applied both in vitro and in vivo. Recently, MRI has been used in tissue engineering to generate spatial maps of tissue relaxation times (T(1), T(2)), water diffusion coefficients, and the stiffness (shear moduli) of developing engineered tissues. In addition, through the use of paramagnetic and superparamagnetic contrast agents, MRI can quantify cell death, assess inflammation, and visualize cell trafficking and gene expression. After tissue implantation MRI can be used to observe the integration of a tissue implant with the surrounding tissues, and to check for early signs of immune rejection. In this review, we describe and evaluate the growing role of MRI in the assessment of tissue engineered constructs. First, we briefly describe the underlying principles of MRI and the expected changes in relaxation times (T(1), T(2)) and the water diffusion coefficient that are the basis for MR contrast in developing tissues. Next, we describe how MRI can be applied to evaluate the tissue engineering of mesenchymal tissues (bone, cartilage, and fat). Finally, we outline how MRI can be used to monitor tissue structure, composition, and function to improve the entire tissue engineering process. PMID- 19134546 TI - Recent developments in microbial fuel cell technologies for sustainable bioenergy. AB - Microbial fuel cells (MFCs) are devices that exploit microbial catabolic activities to generate electricity from a variety of materials, including complex organic waste and renewable biomass. These sources provide MFCs with a great advantage over chemical fuel cells that can utilize only purified reactive fuels (e.g., hydrogen). A developing primary application of MFCs is its use in the production of sustainable bioenergy, e.g., organic waste treatment coupled with electricity generation, although further technical developments are necessary for its practical use. In this article, recent advances in MFC technologies that can become fundamentals for future practical MFC developments are summarized. Results of recent studies suggest that MFCs will be of practical use in the near future and will become a preferred option among sustainable bioenergy processes. PMID- 19134547 TI - A high-throughput and solvent-free method for measurement of natural polyisoprene content in leaves by Fourier transform near infrared spectroscopy. AB - Commercial development of natural polyisoprene from polyisoprene-producing plants requires detailed knowledge on how to select high-polyisoprene-content lines and establish agronomic cultivation methods for achieving maximum polyisoprene yield. This development can be facilitated by a high-throughput quantification method for natural polyisoprene. In this paper, we describe the Fourier transform near infrared spectroscopy (FT-NIR) technique coupled with a partial least squares (PLS) regression model to quantify natural polyisoprene in Eucommia ulmoides leaves. PLS regression models are discussed with respect to linearity, root-mean square error of estimation (RMSEE), and root-mean-square error of prediction (RMSEP). The best PLS regression model was obtained with second derivative NIR spectra in the region between 4000-6000 cm(-1) (R2Y, 0.95; RMSEE, 0.25; RMSEP, 0.37). This is the first report to employ FT-NIR analysis for high throughput and solvent-free quantification of natural polyisoprene in leaves. PMID- 19134548 TI - Comparative characterization of L-lactic acid-producing thermotolerant Rhizopus fungi. AB - Acid-producing Rhizopus fungi from loog-pang, a traditional Thai fermented food, was screened to investigate its potential for use in industrial lactic acid production from starch. A thermotolerant strain, TISTR 3518, was isolated and characterized by its morphological, physiological, genetic and fermentation properties, and compared with its mesophilic isolates, TISTR 3514 and TISTR 3523. TISTR 3518 was characterized by shorter sporangiophores and smaller sporangia than the other isolates; however, apparent differences between the mesophilic isolates and the strain could not be clarified. Moreover, TISTR 3518 grew at 45 degrees C, whereas the others did not. The three isolates showed different profiles of oligosaccharide assimilation and organic acid production. Their rDNA ITS sequences indicated that TISTR 3518 is a strain of Rhizopus microsporus, and TISTR 3514 and TISTR 3523 are strains of Rhizopus oryzae. TISTR 3523 and TISTR 3518 mainly formed L-lactic acid from glucose, while TISTR 3514 primarily formed fumaric acid. Under thermotolerant conditions, R. microsporus TISTR 3518 showed higher glucoamylase activity than the others, suggesting this enzyme from TISTR 3518 is more thermostable than that from TISTR 3523. The strain formed higher amounts of L-lactic acid from starch at 40 degrees C compared to R. oryzae TISTR 3523. This is the first report on the production of optically active L-lactic acid from starch by a thermotolerant fungus and could potentially provide a good tool for transforming biomass resources to chemical materials. PMID- 19134549 TI - Effect of transforming growth factor-beta1 on morphological characteristics relating to migration and differentiation of rabbit chondrocytes cultured in collagen gels. AB - The influence of transforming growth factor-beta1 (TGFbeta1) on the behavior of rabbit chondrocytes embedded in collagen gels was examined in terms of cell migration and consequent architecture of cell aggregation. In a low-seeding density culture (X(0)=2.0 x 10(5) cells/cm(3)) TGFbeta1 (0-10.0 ng/ml) was added and observed during a 14-d culture period. Stereoscopic observation was performed on 5 d employing the morphology-related parameter of sphericity (S(c)) for individual cells in the gels. The frequency of migrating cells with S(c) less than 0.95 increased in a dose-dependent manner in response to TGFbeta1. Moreover, the frequency of migrating cells in the culture with 10.0 ng/ml TGFbeta1 was 0.32, two times higher than that in the reference culture without TGFbeta1, while the frequency of dividing cells in the same culture was less than half of that in the reference culture. The histological observation of cultured gels on 14 d revealed that the starburst and loose aggregates with the spindle-shaped cells emerged in the TGFbeta1-free culture, accompanying the poor production of collagen type II by the cells. On the other hand, the spherical-shaped cells were observed in the starburst aggregates with rich excretion of collagen type II in the culture with 5.0 ng/ml TGFbeta1. Moreover, the mRNA levels of differentiation marker genes (collagen types I and II) were regulated in accordance with the morphological analyses concerning the cell migration and aggregation in the cultures with and without TGFbeta1. From these results, it was concluded that TGFbeta1 had a culture time-dependent effect on the morphological characteristics relating to the migration and differentiation of the chondrocytes in the collagen gel-embedded cultures seeded at low density, that is, the growth factor promotes cell migration with deteriorated proliferation in the early culture phase, and accelerates the transformation of spindle-shaped cells to spherical-shaped ones in the prolonged culture. PMID- 19134550 TI - High catalase production by Rhizobium radiobacter strain 2-1. AB - To promote the application of catalase for treating wastewater containing hydrogen peroxide, bacteria exhibiting high catalase activity were screened. A bacterium, designated strain 2-1, with high catalase activity was isolated from the wastewater of a beverage factory that uses hydrogen peroxide. Strain 2-1 was identified as Rhizobium radiobacter (formerly known as Agrobacterium tumefaciens) on the basis of both phenotypic and genotypic characterizations. Although some strains of R. radiobacter are known plant pathogens, polymerase chain reaction (PCR) analysis showed that strain 2-1 has no phytopathogenic factor. Compared with a type strain of R. radiobacter, the specific catalase activity of strain 2 1 was approximately 1000-fold. Moreover, Strain 2-1 grew faster and exhibited considerably higher catalase activity than other microorganisms that have been used for industrial catalase production. Strain 2-1 is harmless to humans and the environment and produces catalase efficiently, suggesting that strain 2-1 is a good resource for the mass production of catalase for the treatment of hydrogen peroxide-containing wastewater. PMID- 19134551 TI - Stepwise esterification of phytosterols with conjugated linoleic acid catalyzed by Candida rugosa lipase in solvent-free medium. AB - We conducted a near quantitative esterification of phytosterols from soybean oil deodorizer distillate with conjugated linoleic acid. We used a 1:1 molar ratio of sterols to conjugated linoleic acid. For that matter, stepwise addition of sterols was investigated. Total sterols were divided into several portions and added sequentially to the reaction mixture. Using this methodology, purities of up to 80% steryl esters were obtained that consumed more than 90% of the total conjugated linoleic acid. In addition, the effects of temperature, amount, and stability of lipase were also evaluated. PMID- 19134552 TI - Construction and characterization of single-gene chromosomes in Saccharomyces cerevisiae. AB - Based on a previously developed PCR-mediated chromosome splitting method, a genome engineering technique was developed in haploid Saccharomyces cerevisiae for incorporating any desired chromosomal region into a chromosome that carries a single gene. Based on the viability of cells carrying an essential gene in such a construct, close physical proximity of two telomeres and a centromere does not appear to compromise gene function. Spontaneous loss of constructed single-gene chromosomes during vegetative growth was high (0.2-0.4 per cell division), suggesting the possibility of creating novel cells carrying single-gene chromosomes derived from various chromosomal regions in a variety of combinations by exploiting combinatorial loss. PMID- 19134553 TI - Overexpression of GADD34 enhances production of recombinant human antithrombin III in Chinese hamster ovary cells. AB - To improve the production of recombinant human antithrombin III (AT-III) in Chinese hamster ovary (CHO) cells, the gene encoding growth arrest and DNA damage inducible protein 34 (GADD34), which is a transcription factor involved in the unfolded protein response (UPR), was cloned from CHO-K1 cells. Overexpression of GADD34 significantly enhanced the production of recombinant AT-III in CHO 13D-35D cells. The specific rate of AT-III production in the GADD34-overexpressing CHO 13D-35D cells reached approximately 28 pg/cell/d. After 144 h of incubation, the AT-III concentration in the culture supernatant was approximately 40% higher than that observed in the case of the parental CHO 13D-35D cells. The mRNA expression, specific activity, and fucosylation of AT-III were not affected by GADD34 overexpression. Overexpression of GADD34 is a promising method of improving the production of secreted protein pharmaceuticals in CHO cells. PMID- 19134554 TI - Medium composition suitable for L-lysine production by Methylophilus methylotrophus in fed-batch cultivation. AB - L-Lysine production was investigated in fed-batch fermentation using L-lysine producer of Methylophilus methylotrophus. By the addition of nutrient composition, containing L-methionine, K(2)HPO(4), NaH(2)PO(4), CuSO(4).5aq, MnSO(4).5aq, ZnSO(4).7aq, FeCl(3), MgSO(4).7aq and CaCl(2).2aq, in the feed medium, cell growth could be maintained through the cultivation, and L-lysine production reached to 7.86 g. In addition, the effect of counter ion for NH(4)(+) (Cl(-), SO(4)(2-), glutamate, succinate and citrate) was examined. The result showed that the cell growth in the medium using Cl(-) and glutamate were improved compared with that using SO(4)(2-), succinate and citrate, and L-lysine production in the medium using Cl(-) and glutamate reached to more than 9.0 g. In this experiment, there was a clear correlation between ionic strength and growth rate in the cultivation. In order to examine the influence of ionic strength on growth rate, the activity of enzymes in central metabolic pathway from methanol to pyruvate were assayed using samples at the log-phase and the stationary phase in fed-batch cultivation using (NH(4))(2)SO(4) and (NH(4))Cl as ammonium source. It was found that the higher ionic strength inhibited methanol oxidation activity, which linked to cell growth. In this report, it was revealed that maintaining a relatively low ionic strength had a positive effect on L-lysine production using L-lysine producer of M. methylotrophus. PMID- 19134555 TI - Effect of acetate on nitrite oxidation in mixed-population biofilms. AB - While ammonia oxidation has been widely studied, nitrite oxidation is still not well understood. To study the effect of organics on nitrite oxidation, one control group and four treatment groups were designed with sodium acetate (C) to nitrite (N) ratios of 0, 0.44, 0.88, 4.41, and 8.82. Normal nitrite oxidization reactions were performed when C/N=0, but nitrite oxidization and partial denitrification occurred with low C/N ratios (0.44 or 0.88). At high C/N ratios (4.41 or 8.82), we mainly observed denitrification. In contrast to C/N=0, the nitrite oxidization rate was unaffected when C/N=0.44, but decreased with C/N = 0.88. The structure of bacterial communities varied significantly between autotrophic and mixotrophic conditions. Nitrobacter was hard to detect by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) while heterotrophs and especially denitrifiers were in the majority under mixotrophic conditions. Real-time PCR indicated that the Nitrobacter population decreased from 2.42 x 10(4) to 1.34 x 10(3) 16S rRNA gene copies/ng DNA, while the quantity of denitrifiers obviously increased from 0 to 2.51 x 10(4) nosZ gene copies/ng DNA with an increasing C/N ratio. The degree of denitrification differed between C/N=4.41 and C/N=8.82, as indicated by nitrite-N and nitrate-N curve analysis, as well as by the apparent bacterial community structure. Our findings provide critical insight regarding the relationship between organics and nitrification in biofilms. PMID- 19134557 TI - Breeding of industrial diploid yeast strain with chromosomal integration of multiple beta-glucosidase genes. AB - We constructed a double auxotrophic OC-2 industrial diploid strain of Saccharomyces cerevisiae and introduced 4 copies of cell surface displaying beta glucosidase (BGL) genes into the chromosome. The engineered OC-2 strain showed 5 fold higher BGL activity compared with the yeast carrying 2 copies of BGL gene and directly produced ethanol from cellobiose. PMID- 19134556 TI - Knowledge-based fuzzy adaptive resonance theory and its application to the analysis of gene expression in plants. AB - Gene expression data obtained from DNA microarrays are very useful in revealing the mechanisms that drive life. It is necessary to analyze these data through the use of algorithms, as in clustering and machine-learning. In a previous study, we developed fuzzy adaptive resonance theory (FuzzyART) and applied it to gene expression data, to identify genetic networks. FuzzyART was used as a clustering algorithm that is very suitable for the analysis of biological data; however, although FuzzyART is very useful in the analysis of dozens of gene expression profiles, it is difficult to apply this method to thousands of gene expression profiles, owing to inherent category proliferation and long calculation time. In the present study, we developed a knowledge-based FuzzyART (KB-FuzzyART) to mitigate these problems. We first constructed a gene list-1 from the gene database of Arabidopsis thaliana as knowledge for KB-FuzzyART, because KB FuzzyART requires any knowledge as input. This method was applied to gene expression data obtained via the microarray analysis of A. thaliana, to identify the downstream genes of ASYMMETRIC LEAVES1 (AS1) and ASYMMETRIC LEAVES2 (AS2), both of which are involved in leaf development. The results of the analysis using KB-FuzzyART showed that the KNAT6 and YABBY5 (YAB5) genes are candidates for downstream factors, after a short calculation time for analysis. These results suggest that our gene list-1 is a very useful database for analyzing the expression profiles of genes that are related to the development of A. thaliana; they also suggest that the KB-FuzzyART has the high potential to function as a new method by which one can select candidate genes from thousands of genes, using gene expression data on mutant strains. PMID- 19134558 TI - Retroviral gene transduction into chicken embryo gonads through blood circulation. AB - A retroviral vector was injected into the heart of developing chicken embryos to determine the best timing for effective gene transduction into the gonads. Transduction efficiency and transgene expression were both highest when the viral solution was injected into embryos at stages 14-15 (Hamburger-Hamilton stages), in which primordial germ cells (PGCs) can be found in the blood circulation. This study shows that PGCs in the blood are important targets for gene transfer into the gonads using retroviral vectors. PMID- 19134559 TI - Enhanced release of chitosanase from Streptomyces griseus through direct interaction of liposome with cell membrane under heat stress. AB - A direct interaction of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes with membrane of Streptomyces griseus cell under the heat stress at 41 degrees C increased the chitosanase production and release to 2.2 times higher than that at 37 degrees C without the POPC liposomes. Amount of chitosanase released across the lipid mimicking cell membrane (LMCM) liposome under a heat at 41 degrees C in the presence of POPC liposomes was 17% of initially-entrapped chitosanase while it was only 1% in the absence of POPC liposomes, even under a heat stress at 41 degrees C, clearly showing the importance of the direct interaction between membrane and membrane. PMID- 19134560 TI - Three dimensional finite element analyses of oral structures by computerized tomography. AB - Our aim was to document the benefits of three dimensional finite element model generations from computed tomography data as well as the realistic creation of all oral structures in a patient. The stresses resulting from the applied load in our study did not exceed the structure limitations, suggesting a clinically acceptable physiological condition. PMID- 19134561 TI - Vitrification of corneal endothelial cells in a monolayer. AB - An in vitro model was developed for bovine natural corneal endothelia. The cells were cultured to a confluent monolayer and vitrified using 25% (w/w) 1,2 propanediol-35% (w/w) trehalose as cryoprotective agents. Approximately, 61.3% of the cells were viable using the protocol. PMID- 19134996 TI - A review of the otological aspects of whiplash injury. AB - Approximately 10% of patients who have suffered with whiplash injury will develop otological symptoms such as tinnitus, deafness and vertigo. Some of these are purely subjective symptoms; nevertheless, for the majority there are specific tests that can be undertaken. These tests can quantify the extent and severity of the symptoms as well as provide guidance as to the correct rehabilitation pathway. This article reviews the body of literature relating to the otological aspects of whiplash injury and gives an overview for medical and legal professionals. PMID- 19134997 TI - Sternal index: Is it a reliable indicator of sex in the Maharashtrian population of India? AB - Sex determination is an important initial step in forensic investigation of unknown human skeletal remains. The present analysis is an attempt to determine the sex of the sternum using sternal index as a parameter in a Maharashtrian population of India. Sternal index is calculated in 115 adult sternums of confirmed sex (75 male and 40 female) for sexual dimorphism. The sternal index is derived by dividing the length of manubrium by the length of mesosternum and multiplying it by 100. Although the sternal index is found to be significantly higher in females, the analysis suggests that sex determination cannot be made conclusively from the sternal index in the Maharashtra region of western India. The study further concludes that the applicability of Hyrtl's law in sex determination is limited. PMID- 19134998 TI - Qualitative and quantitative EEG abnormalities in violent offenders with antisocial personality disorder. AB - Resting eyes closed electroencephalogram was studied in a group of violent offenders evaluated at Psychiatric Department of the Legal Medicine Institute in Cuba (18 with antisocial personality disorder, ASPD, and 10 without psychiatric diagnosis). Characteristics of the EEG visual inspection and the use of frequency domain quantitative analysis techniques (narrow band spectral parameters) are described. Both groups were compared to Cuban normative database. High incidences of electroencephalographic abnormalities were found in both groups of violent offenders. The most frequent were: electrogenesis alterations, attenuated alpha rhythm and theta and delta activities increase in the frontal lobe. In the quantitative analysis theta and delta frequencies were increased and alpha activity was decreased in both groups. Differences appear for the topographical patterns present in subjects of both groups. EEG abnormalities were more severe in ASPD than in control group. Results suggest that EEG abnormalities in violent offenders should reflect aspects of brain dysfunction related to antisocial behaviour. PMID- 19134999 TI - Prevalence of dihydrocodeine in hydrocodone positive postmortem specimens. AB - Hydrocodone (HC) has received renewed interest in the US due to reported increases in opiate related deaths involving psychotherapeutic drugs. The relative contribution of dihydrocodeine (DHC) in these deaths is unknown since little testing of this compound is performed. The objective of the study was to determine the prevalence of DHC in HC positive decedents and report the range of concentrations detected in these cases in order to evaluate the potential role of DHC in the deaths and determine the usefulness of including this analyte in opioid testing protocols. Specimens were assayed by liquid-liquid or solid phase extraction followed by gas chromatography/mass spectrometry operated in the selected ion monitoring mode. A multipoint calibration was utilized in the linear range 2-600ng/mL. Accuracy for HC, DHC and hydromorphone (HM) was 101-106% and between day precision at 160ng/mL between 7% and 11%. One hundred and thirty six cases were identified with the majority male (62%) and white (83%). A search of HC positive cases identified 64 with DHC (47%). The range of HC concentrations was 9-3039ng/mL heart blood (n=43) and 42-12353ng/mL urine (n=21). DHC concentrations in these cases ranged 3-243ng/mL in heart blood and 5-1842ng/mL in urine. DHC/HC ratios ranged 0.00(7)-2.90 in blood (n=43), and 0.01-5.04 in urine (n=21) with 16% and 24% of these cases with ratios >0.50, respectively. HM was detected in only 9 HC cases with the majority positive in urine. PMID- 19135000 TI - Limitations of the mandibular canine index in sex assessment. AB - Measuring teeth is a useful adjunct in sex assessment. Canines, in particular, have the greatest degree of sexual dimorphism, resist disease and survive postmortem trauma, rendering them highly valuable in identification. Hence, their exclusive use in odontometric sex assessment using the Mandibular Canine Index (MCI) has been advocated before. The MCI is derived as the ratio of the mesiodistal (MD) dimension of canines and the inter-canine arch width. This study has tested the use of the MCI in assessing sex on a sample from Nepal and compared its accuracy to that of absolute canine measurements. Measurements were obtained from one hundred-and-seventeen dental stone casts that belonged to 63 males and 54 females, all young adults in the age-group 19-28 years. Independent samples t-test revealed no significant sexual dimorphism in the MCI. In addition, discriminant analysis of the MCI also had poor ability to differentiate the sexes. In contrast, the absolute canine measurements revealed statistically significant male-female difference and superior ability to differentiate sex using discriminant analysis. The poor ability of the MCI in sex assessment is attributed to it being a relative value-it is obtained as the ratio of two absolute measurements (MD dimension of canines and inter-canine arch width) and does not reflect sex differences that exist in the absolute measurements per se. PMID- 19135001 TI - Filicide and fatal abuse in Japan, 1994-2005: temporal trends and regional distribution. AB - To describe the recent filicide features in Japan, data were collected from newspaper databases between 1994 and 2005 and compared with several official statistics. From this dataset, 933 cases and 1084 victims under age 15 were identified. Fatal abuse cases were most prevalent (309 cases, or 33.1%), followed by filicide-suicide cases (303, or 32.5%) and unwanted child cases (225, or 24.1%). Infants under the age of one were most at risk (filicide rate: 2.72 per 100,000 for the same aged infants). Annual filicide numbers showed an upward trend between 1994 and 2001 and the average filicide rate per year was 0.42 per 100,000 for children under age 15. Both the number of cases over the period and the filicide rate per year by prefecture ranged widely. Furthermore, the annual filicide rates over the period were strongly correlated with both suicide rates and unemployment rates for the general population (r=.884 and .926, respectively; p<.001 for both). The measure of filicide per region and the predominating category in a prefecture provides direction for filicide prevention. Result suggests that recent socioeconomic situations have a significant effect on not only filicide-suicide but also the overall filicide rate in Japan. PMID- 19135002 TI - A five-year survey for dental malpractice claims in Tehran, Iran. AB - OBJECTIVES: Dentists, like other doctors, can face punitive and legal consequences if patients are not satisfied with the dental treatment. The purpose of this study is to provide a database for dental malpractice claims in Tehran. METHODS: We conducted a retrospective study of dental malpractice claims In Tehran, between 2002 and 2006, based on the decisions of expert committees in medical malpractice cases by Tehran's Legal Medicine Organization and Islamic Republic of Iran's Medical Council. RESULTS: During these 5 years, 412 decisions related to dental malpractice were made. The majority of complaints were in fixed prosthodontics and oral surgery and also most of them concerned the private sector. Most of the cases were against general dentists. In the 56.7% of clinical cases and 40% of non-clinical cases of malpractice claims, dentists were found faulty. CONCLUSIONS: Like all other medical staff, dentists are under the obligation to comply with the legal rules in the country they practice. They also have to consider ethical principles as well as the acceptable standards and protocols of diagnosis and treatment. These data can alert them to the need for greater care and ethical professionalism when treating their patients. PMID- 19135003 TI - Applied forensic epidemiology: the Bayesian evaluation of forensic evidence in vehicular homicide investigation. AB - The comparative weighting of evidence in a criminal case can be a complicated task when the relevance or meaning of the evidence is disputed. An example of this complexity in seen in vehicular homicide investigations in which the identity of the driver (and thus the guilty party) is not clear. The discipline of Forensic Epidemiology, including the appropriate application of Bayes' Theorem (Bayes' Law) provides a systematic framework to bring clarity to the evaluation of such matters. Bayes' is a useful tool for the conditioning and quantification of probabilities associated with evidence in a vehicular homicide investigation. The authors present a case study in the application of Bayes' Theorem to the facts in a vehicular homicide investigation. An initial analysis of the crash dynamics in comparison with the injury pattern and ejection status of the surviving occupant versus that of the decedent suggested that the survivor was the driver. The results of the analysis were used as tests for guilt, with estimated true and false positive rates, which then formed the basis for a Bayesian calculation of the posterior probability of the survivor's guilt given the evidence. As a result of the Bayesian analysis described herein, it was determined that the survivor was 19 times more likely to have been the driver, in comparison with the decedent. This ratio far exceeded the suggested threshold of 10:1 for establishing the guilt of the survivor beyond a reasonable doubt. When used properly, Bayes' Theorem can offer definitive insight in the investigation and prosecution of vehicular homicide cases. PMID- 19135004 TI - Medical confidentiality versus disclosure: Ethical and legal dilemmas. AB - A case is described of a fifty year old single man who made disclosures about criminal sexual practices during a psychiatric assessment. In common practice with other professional men, a doctor is under a duty not to disclose, without the consent of his patient, information which he has gained in his professional capacity other than in exceptional circumstances. We discuss the ethical and legal considerations surrounding issues of medical confidentiality and the dilemma that sometimes face clinicians, when they feel obliged, in the public interest, to disclose information they have gained in confidence. Breach of confidences can have deleterious consequences; particularly for the doctor patient relationship, but failure to disclose in some situations could have serious implications for the well-being of the wider society. Doctors should be aware of the basic principles of confidentiality and the ethical and legal framework around which they are built. PMID- 19135005 TI - Two death cases originating from supplementary heater in the cabins of parked trucks. AB - A supplementary heater can be mounted in trucks or other transportation vehicles and used to heat the air in the cabin independent of the engine of the vehicle. This apparatus works with diesel fuel or gasoline. Combustion products burning in the pre-combustion chamber heat the fins of the engine. The air passing through the fins is heated and is transferred into the cabin. The malfunction of such an apparatus may be the cause of carbon monoxide (CO) poisoning or fire. In this study, we report two cases in which drivers died while asleep in the cabins of parked trucks. In the first case, a 43-year-old man died because of CO poisoning originating from a broken supplementary heater. In the second case, a 48-year-old man died owing to a fire that resulted from the burning of upholstery cloths in the truck cabin, and which was caused by a supplementary heater set at very high temperatures. In both cases, it was determined by a technical expert that the CO poisoning in the first case and the fire in the second case were caused by the supplementary heater in the trucks. It is emphasized in this article that the supplementary heater in the truck might be the cause of mortal CO poisoning as well as be responsible for fires in the cabins of the trucks. It aims to highlight that a detailed investigation of supplementary heaters at the death scene is required for such death cases in the trucks. PMID- 19135006 TI - Acute haemorrhagic pancreatitis - A case of sudden death. AB - Sudden death in young individuals is relatively uncommon and acute haemorrhagic pancreatitis as a cause of sudden and unexpected death is a rare phenomenon. A case of sudden death due to acute haemorrhagic pancreatitis is reported in a young doctor who ignored the radiating pain of acute pancreatitis for gastritis, and resorted to self-medication. The condition was discovered only at autopsy. PMID- 19135007 TI - Analysis of the high-risk age group of suicide in South Korea-Comparison of Japan and South Korea. PMID- 19135008 TI - A letter from America: The ghost of Dr. Griggs' 2008;15(1):7-15. PMID- 19135009 TI - Emergency nursing - home and away. PMID- 19135010 TI - A descriptive study on emergency department doctors' and nurses' knowledge and attitudes concerning substance use and substance users. AB - AIM: The aim of this study was to determine emergency department doctors' and nurses' knowledge and attitudes regarding problematic substance use and substance users. METHODS: Data were collected using an adapted survey questionnaire and the Substance Abuse Attitude Survey (SAAS). By means of convenience/opportunistic sampling all emergency department doctors and nurses (N=145) working in three university teaching hospitals in Ireland were asked to fill out the knowledge and attitudes questionnaire. RESULTS: A response rate of 46%, N=66 was achieved. Results indicate that participants' current level of knowledge about alcohol and drug misuse in general, is satisfactory. A particular knowledge deficit in relation to intervention strategies, and other substances was identified. The majority of participants have never received any specific training regarding substance use and this suggests that substance using patients are managed inadequately. The SAAS results indicate that participants exhibited near-optimal attitudes for constructive working with substance using patients. CONCLUSION: There is an urgent need for further in-service training programmes and the development of standard protocols/guidelines for the identification and management of substance using patients who present in the emergency department. PMID- 19135011 TI - Patient participation in emergency care - a phenomenographic study based on patients' lived experience. AB - International guidelines promote patient participation in health care. When patients participate in their care they experience greater satisfaction. Studies have shown that patients in emergency departments express dissatisfaction with their care, and it was therefore important to study how patients understand and conceptualize their participation. The aim of this study was to describe patients' qualitatively different conceptions of patient participation in their care in an emergency department. Based on a lifeworld perspective, nine interviews were performed with patients in an emergency department. The phenomenographic analysis shows that participation by patients means contact with the emergency department staff in three categories of conceptions: being acknowledged; struggling to become involved; and having a clear space. The different conceptions of patient participation give us a deeper understanding of how patients may experience their care, and this result may provide a foundation for developing nursing practice and the quality of health care in line with international guidelines. PMID- 19135012 TI - Attitudes and beliefs of emergency department staff regarding alcohol-related presentations. AB - OBJECTIVE: This study examined emergency department (ED) staff attitudes and beliefs about alcohol-related ED presentations in order to recommend improved detection and brief intervention strategies. METHODS: The survey was conducted at two inner-Sydney hospital EDs in 2006 to explore ED clinical staff's attitudes, current practice and barriers for managing alcohol-related ED presentations. The sample included N=78 ED staff (54% nurses, 46% doctors), representing a 30% response rate. RESULTS: Management of alcohol-related problems was not routine among ED staff, with only 5% usually formally screening for alcohol problems, only 16% usually conducting brief interventions, and only 27% usually providing a referral to specialist treatment services. Over 85% of ED staff indicated that lack of patient motivation made providing alcohol interventions very difficult. Significant predictors of good self-reported practice among ED staff for patients with alcohol problems included: being a doctor, being confident and having a sense of responsibility towards managing patients with alcohol-related problems. CONCLUSIONS: This study reported that many staff lack the confidence or sense of clinical responsibility to fully and appropriately manage ED patients with alcohol-related problems. ED staff appear to require additional training, resources and support to enhance their management of patients with alcohol related problems. PMID- 19135013 TI - Translating change: the development of a person-centred triage training programme for emergency nurses. AB - Within health care, there has been a change in practice from an illness orientated service to one that is more health-focused and person-centred. The concept of person-centredness is frequently espoused by practitioners as being not only a desirable, but a necessary element of health care provision. Indeed, nationally and internationally, person-centred care has underpinned many healthcare documents and policies. Person-centred practice focuses on providing care, utilising a variety of processes that operationalise person-centred nursing and include working with patients' beliefs and values, engagement, presence, sharing decision-making and providing for physical needs. In the field of emergency nursing, the incorporation of person-centred care and its holistic foundation may require a significant shift in practice. There is evidence to suggest that emergency nurses view their role as one, which is predominantly concerned with providing urgent physical care, rather than one, which espouses the theories of holistic healthcare. To this extent, being person-centred in the context of emergency care, requires the nurse to move beyond the traditional notions of his/her role and to embrace the more holistic aspects of patient care. The aim of this article is to critically analyse how a change in nurse-led triage training in one Irish Emergency Department facilitated an improved person-centred approach in practice. PMID- 19135014 TI - An investigation comparing the Oxford Chair Technique with the traditional methods of glenohumeral dislocation reduction currently implemented. AB - This paper reports on the findings of a quantitative, retrospective service evaluation. It compared a new, conscious, sedation-free technique to reduce anterior glenohumeral dislocations with the traditional methods of reduction (TMR) currently implemented. OBJECTIVES: The first objective for this study was to examine whether a new method, used at an Emergency Department (ED) in the south of England, of glenohumeral dislocation reduction, called the Oxford Chair Technique (OCT) can reduce anterior glenohumeral dislocations and to compare the success rates of reduction with the TMR. Secondary objectives examined how the OCT compared, using four set factors, against the TMR currently used. The fundamental comparisons being analysed was in terms of time taken for treatments. METHODS: A retrospective service evaluation was performed using data collected from the Patient Administration System (PAS) software and the Picture Archiving and Communications System (PACS) on patients who presented to an ED in the South of England with an anterior glenohumeral dislocation between March 2005 and August 2006. RESULTS: Of the 61 cases in which the OCT was used over the sampling period, 38 attempts to reduce the dislocation were successful, with 23 being unsuccessful. An overall success rate of 62% was obtained with the OCT. There were no complications reported with the OCT. Statistically significant differences were found with the mean time from arrival to discharge (OCT 141 v. TMR 254 min, p<0.001); mean time between the first diagnostic X-ray and post reduction X-ray (OCT 70 v. TMR 102 min, p<0.003) and the mean time between the post reduction X-ray and discharge (OCT 51 v. TMR 119 min, p<0.001). No patients treated with the OCT required conscious sedation compared to 90% of patients treated with TMR (p<0.001). Only 38% of patients treated with the OCT had morphine administered compared to 90% of patients treated with TMR (p<0.001). CONCLUSIONS: Overall, the success rate for reduction using the OCT was good in view of this being a new treatment to the ED. While the OCT had a lower success rate of reduction compared to TMR, when reduction was successful using the OCT, there were overwhelming time-saving benefits to the patient with subsequent logistical benefits to the ED. The use of conscious sedation and morphine was also significantly less for the OCT group compared to TMR. PMID- 19135015 TI - Physical environment of the home and adolescent injury risk. AB - OBJECTIVES: The home environment is one of the most significant contextual factors that contributes to young children's unintentional injury risk, but there are very limited data concerning risks present in the homes of adolescents. This study was designed to offer descriptive data on aspects of the home physical environment that might contribute to adolescent injury risk in the United States. METHODS: A diverse sample of 42 adolescents ages 14-16 participated. Researchers completed an inspection of the adolescents' home, searching for various safety related hazards. RESULTS: Homes tended to be safe in some domains (e.g., presence of smoke detectors), but had substantial risk in other domains. For example, over 90% of homes were without functioning carbon monoxide detectors; 29% had unlocked firearms present; 21% had exposed electrical cords; and 31% had alcohol present and unlocked. CONCLUSIONS: Although residential environment risks are viewed to be most concerning for very young children, over 30% of fatal adolescent injuries occur in the home. Results suggest there are substantial risks to adolescent safety in the home environment. PMID- 19135016 TI - Incidence and severity of head and neck injuries in victims of road traffic crashes: In an economically developed country. AB - BACKGROUND: Head and neck injuries following the road traffic crashes (RTCs) are the most common cause of morbidity and mortality in most developed and developing countries and may also result in temporary or permanent disability. OBJECTIVE: The aim of this study was to determine the incidence pattern of head and neck injuries, investigate its trend and identify the severity of injuries involved with road traffic crashes (RTCs) during the period 2001-2006. DESIGN: This is a retrospective descriptive hospital based study. SETTING: The patients with head and neck injuries were seen and treated in the Accident and Emergency Department of the Hamad General Hospital and other Trauma Centers of the Hamad Medical Corporation following the road traffic crashes during the period 2001-2006. PATIENTS AND METHODS: This study is a retrospective analysis of 6709 patients attended and treated at the Accident and Emergency and Trauma centers for head and neck injuries over a 6 year period. Head and neck injuries were determined according to the ICD 10 criteria. Of these, 3013 drivers, 2502 passengers, 704 pedestrians and 490 two wheel riders (motor bike and cyclists). Details of all the road traffic crash patients were compiled in the database of the Emergency Medical Services (EMS), and the data of patients with head and neck injuries were extracted from this database. RESULTS: A total of 6709 patients with head and neck injuries was reported during the study period. Majority of the victims were non-Qataris (68.7%), men (85.9%) and in the age group 20-44 years (68.5%). There were statistical significant differences in relation to age, nationality, gender, and accident during week ends for head and neck injuries (p<0.001). The male to female ratio for head and neck injury was 6.1:1. There was a disproportionately higher incidence of accidents during weekends (27.8%). Majority of the patients had mild injury (87.2%), followed by moderate (7.3%) and severe (5.5%). The highest frequency of head injury was among the young adults 20-44 years (68.5%). There was a remarkable increase in the incidence rate of head and neck injuries per 10,000 population in the year 2005 (18.2) compared to previous years and declined slightly in the year 2006 (17.1). Overall, the incidence of head and neck injuries from road traffic crashes are increasing. CONCLUSION: The present study findings provided an overview of head and neck injuries in Qatar from road traffic crashes. The incidence of head and neck injuries is still very high in Qatar, but the severity of injury was mild in most of the victims. The findings of the study highlighted the need for taking urgent steps for safety of people especially drivers and passengers. PMID- 19135017 TI - The role of nurses working in emergency and critical care environments in the prevention of intravascular catheter-related bloodstream infections. AB - Intravascular catheter-related infections are a major problem in healthcare. This review provides up-to-date guidance of evidence-based recommendations for the prevention of intravascular catheter-related infections with special focus on strategies relevant for nurses working in emergency and critical care environments or practitioners responsible for surveillance and control of infections. The review concludes by providing a range of approaches advocated for: (i) translating guidelines to the needs and expectations of emergency and critical care nurses, and (ii) increasing the chance of successful implementation and compliance with these recommendations. PMID- 19135018 TI - Ketamine associated cystitis - a case report. PMID- 19135019 TI - Case review: a 28-year-old Korean man with Irukandji syndrome. PMID- 19135020 TI - Haplotype identification within Trypanosoma cruzi I in Colombian isolates from several reservoirs, vectors and humans. AB - Genetic variability in the Trypanosoma cruzi I group has recently been revealed in Colombian isolates from humans, reservoirs and vectors. Genomic rearrangements and the polymorphic regions in taxonomic markers, such as the miniexon gene, have led to the development of molecular tools to identify phylogenetic haplotypes in T. cruzi isolates. From genetic polymorphisms found in T. cruzi I isolates, they have been classified into four haplotypes according to their epidemiologic transmission cycles. Haplotype Ia is associated with domestic isolates, from Rhodnius prolixus; haplotype Ib, with the domestic and peridomestic cycle, mainly associated with Triatoma dimidiata; haplotype Ic is a poorly characterized group, which has been associated with the peridomestic cycle; and haplotype Id has been related to the sylvatic cycle. In order to demonstrate that the circulating T. cruzi I isolates in Colombia can be classified in the four proposed haplotypes, specific primers were designed on polymorphic regions of the miniexon gene's intergenic sequences. This set of primers allowed the molecular characterization of 33 Colombian isolates, classifying them into three of the four proposed haplotypes (Ia, Ib and Id). Results obtained from maximum parsimony and maximum likelihood-based phylogenetic analyses correlated with the molecular classification of the isolates and their transmission cycles. This study brings insights into the Chagas disease epidemiology and the parasite's transmission dynamics. PMID- 19135021 TI - Detergent screening of a G-protein-coupled receptor using serial and array biosensor technologies. AB - We describe the benefits and limitations of two biosensor approaches for screening solubilization conditions for G-protein-coupled receptors (GPCRs). Assays designed for a serial processing instrument (Biacore 2000/3000/T100) and an array platform (Biacore Flexchip) were used to examine how effectively 96 different detergents solubilized the chemokine receptor CCR5 while maintaining its binding activity for a conformationally sensitive Fab (2D7). Using the serial processing instrument, we were able to analyze three samples in each 30-min binding cycle, thereby requiring approximately 24h to screen an entire 96-well plate of conditions. In-line capturing allowed us to normalize the 2D7 binding responses for different receptor capture levels. In contrast, with the array system, we could characterize the effects of all 96 detergents simultaneously, completing the assay in less than 1h. But the current array technology requires that we capture the GPCR preparations off-line, making it more challenging to normalize for receptor capture levels. Also, the array platform is less sensitive than the serial platforms, thereby limiting the size of the analyte to larger molecules (>5000Da). Overall, the two approaches proved to be highly complementary; both assays identified identical detergents that produced active solubilized CCR5 as well as those detergents that either were ineffective solubilizers or inactivated the receptor. PMID- 19135022 TI - Quantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - The physiological response to small molecules (secondary messengers) is the outcome of a delicate equilibrium between biosynthesis and degradation of the signal. Cyclic diguanosine monophosphate (c-di-GMP) is a novel secondary messenger present in many bacteria. It has a complex cellular metabolism whereby usually more than one enzyme synthesizing and degrading c-di-GMP is encoded by a bacterial genome. To assess the in vivo conditions of c-di-GMP signaling, we developed a high-performance liquid chromatography (HPLC)-mass spectrometry-based method to detect c-di-GMP with high sensitivity and to quantify the c-di-GMP concentration in the bacterial cell as described here in detail. We successfully used the methodology to determine and compare the c-di-GMP concentrations in bacterial species such as Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and Vibrio cholerae. We describe the use of the methodology to assess the change in c-di-GMP concentration during the growth phase and the contribution of a point mutation in S. typhimurium to the overall cellular c-di-GMP concentration. PMID- 19135023 TI - A nonradioactive high-throughput assay for screening and characterization of adenylation domains for nonribosomal peptide combinatorial biosynthesis. AB - Adenylation domains are critical enzymes that dictate the identity of the amino acid building blocks to be incorporated during nonribosomal peptide (NRP) biosynthesis. NRPs display a wide range of biological activities and are some of the most important drugs currently used in clinics. Traditionally, activity of adenylation domains has been measured by radioactive ATP-[32P]pyrophosphate (PP(i)) exchange assays. To identify adenylation domains for future combinatorial production of novel NRPs as potential drugs, we report a convenient high throughput nonradioactive method to measure activity of these enzymes. In our assay, malachite green is used to measure orthophosphate (P(i)) concentrations after degradation by inorganic pyrophosphatase of the PP(i) released during aminoacyl-AMP formation by action of the adenylation domains. The assay is quantitative, accurate, and robust, and it can be performed in 96- and 384-well plate formats. The performance of our assay was tested by using NcpB-A(4), one of the seven adenylation domains involved in nostocyclopeptide biosynthesis. The kinetics of pyrophosphate release monitored by this method are much slower than those measured by a traditional ATP-[32P]PP(i) exchange assay. This observation indicates that the formation of the adenylated amino acid and its release are the rate-limiting steps during the catalytic turnover. PMID- 19135024 TI - 3-Carboxy-6-chloro-7-hydroxycoumarin: a highly fluorescent, water-soluble violet excitable dye for cell analysis. AB - In our search for new violet-excitable dyes with improved photophysical and photochemical properties, we examined several halogen-substituted hydroxycoumarins and found that chlorinated derivatives are at least as bright as their fluorinated analogs. A monochlorinated hydroxycoumarin was found to have a high quantum yield (approximately 0.98), and human leucocyte-specific monoclonal antibodies (CD3, CD4, and CD45) conjugated with this dye exhibited reliable performance in flow cytometry assays. Additional studies were performed, with BD Horizon V450-antibody conjugates being included in eight-color cocktails aimed at subsetting lymphocytes and myeloid cells. Such cocktails can frequently be unstable due to the tendency of one or more components to lose structural integrity, photobleach, or develop unwanted affinities for another component. However, the cocktails employed in this study enabled several different applications to be run and established that multicolor reagent mixtures containing V450-antibody conjugates are functional and stable. PMID- 19135025 TI - (-)-Epigallocatechin-3-gallate induces up-regulation of Th1 and Th2 cytokine genes in Jurkat T cells. AB - In the present study, we found that (-)-epigallocatechin-3-gallate (EGCG) significantly up-regulated the mRNA expression of the Th1/Th2 cytokines including IL-2, IFN-gamma, IL-5 and IL-13 in Jurkat T cells. The EGCG-induced mRNA up regulation of IL-2 and IL-5 was predominantly affected by the extracellular signal-regulated protein kinase (ERK) signalling, whereas IL-13 gene expression, the most responsive to the EGCG treatment, was dependent on neither ERK nor c-jun NH(2)-terminal kinase (JNK) signalling. IFN-gamma gene expression was partially mitigated by both inhibitors of the ERK and JNK pathways. Furthermore, catalase significantly attenuated the intracellular peroxide production, phosphorylation of ERK and JNK, and all cytokine gene expressions induced by EGCG. In addition, physiologically relevant concentrations of both EGCG and H(2)O(2)-induced up regulation of IL-5 gene expression. Our findings provide biological evidence that EGCG induces Th1/Th2 cytokine mRNA expression via H(2)O(2) production followed by activation of ERK or JNK in Jurkat T cells. PMID- 19135026 TI - The sea urchin embryo: a model to study Alzheimer's beta amyloid induced toxicity. AB - Alzheimer's disease (AD) is the most common form of dementia. The cause of AD is closely related to the accumulation of amyloid beta peptide in the neuritic plaques. The use of animal model systems represents a good strategy to elucidate the molecular mechanism behind the development of this pathology. Here we use the Paracentrotus lividus embryo to identify molecules and pathways that can be involved in the degenerative process. As a first step, we identified the presence of an antigen related to the human APP, called PlAPP. This antigen, after gastrula stage, is processed producing a polypeptide of about 10kDa. By immunohistochemistry we localized the PlAPP antigen in some serotonin expressing cells. Similarly, after 48 or 96h incubation, a recombinant beta-amyloid peptide, rAbeta42, accumulates around the intestinal tube and oesophagus. In addition, incubation of sea urchin embryos with two different solutions rich in oligomers and fibrillar aggregates of rAbeta42 induce activation of apoptosis as detected by TUNEL assay. Moreover, we demonstrate that aggregates induce apoptosis by extrinsic pathway activation, whereas oligomers induce apoptosis both by extrinsic and intrinsic pathway activation. Utilizing an apoptotic inhibitor, caspases activation was offset and morphological damage rescued. Taken together all these observations suggest that the sea urchin may be a simple and suitable model to characterize the mechanism underlining the cytotoxicity of Abeta42. PMID- 19135027 TI - Expression of sarco/endoplasmic reticulum Ca(2+) ATPase (SERCA) 3 proteins in two major conformational states in native human cell membranes. AB - The SERCA family includes 3 genes (SERCA1-3), each of which giving rise to various isoforms. To date, detailed structural data is only available for the SERCA1a isoform. Here, limited trypsinolysis of either human platelet membranes or recombinant SERCA3a in HEK-293 cells followed by Western blotting using antibodies covering different regions of the SERCA3(a) protein revealed two, kinetically distinct, Early (ETF) and Late (LTF) Tryptic Fragmentations. The ETF uses many tryptic sites while the LTF uses a unique tryptic site. Using site directed mutagenesis: i) Arg(334), Arg(396) and Arg(638) were directly assigned to the ETF and ii) Arg(198) was assigned as the only tryptic site to the LTF. Arg(671), Lys(712)/Lys(713) and Lys(728) were also found to modulate the ETF. SERCA inhibitors Tg and tBHQ induced modest inhibition of the ETF. In contrast, the addition of CaCl(2), EGTA or AlF(4)(-) strikingly modified the ETF without any effect on the LTF. Trypsinolysis of the other recombinant SERCA3b-3f isoforms revealed: i) same ETF and LTF as SERCA3a, with variations of the length of the C terminal fragments; ii) Arg(1002) as an additional tryptic site in SERCA3b-3e isoforms. Taken together, the two distinct SERCA3 fragmentation profiles sign the co-expression of SERCA3 proteins in two conformational states in cell membranes. PMID- 19135028 TI - Variant late infantile ceroid lipofuscinoses associated with novel mutations in CLN6. AB - The neuronal ceroid lipofuscinoses (NCL) are heterogeneous neurodegenerative disorders with typical autofluorescence material stored in tissues. Ten clinical NCL forms and eight causative genes are known. Mutations in CLN6 have been reported in roughly 30 patients, mostly in association with the variant late infantile NCL (v-LINCL) phenotype. We screened CLN6 in 30 children from a cohort of 53 v-LINCL cases and revised their clinical and ultrastructural features. We detected 11 mutations, eight of which are novel, all predicting a direct impairing of the putative gene function. No clear-cut genotype-phenotype correlations were observed, with inter- and intra-familial variability evident for few recurrent mutations. Ultrastructural findings were suggestive of an impaired regulation of the autophagic vacuoles turnover. While expanding the array of CLN6 mutations, we showed that more than half of our v-LINCL cases lack a DNA confirmation and further molecular etiologies are to be searched. PMID- 19135029 TI - Effects of iron oxide nanoparticles on cardiac differentiation of embryonic stem cells. AB - The therapeutic potential of transplantation of embryonic stem cells (ESCs) in animal model of myocardial infarction has been consistently demonstrated. The development of superparamagnetic iron oxide (SPIO) nanoparticles labeling and cardiac magnetic resonance imaging (MRI) have been increasingly used to track the migration of transplanted cells in vivo allowing cell fate determination. However, the impact of SPIO- labeling on cell phenotype and cardiac differentiation capacity of ESCs remains unclear. In this study, we demonstrated that ESCs labeled with SPIO compared to their unlabeled counterparts had similar cardiogenic capacity, and SPIO-labeling did not affect calcium-handling property of ESC-derived cardiomyocytes. Moreover, transplantation of SPIO-labeled ESCs via direct intra-myocardial injection to infarct myocardium resulted in significant improvement in heart function. These findings demonstrated the feasibility of in vivo ESC tracking using SPIO-labeling and cardiac MRI without affecting the cardiac differentiation potential and functional properties of ESCs. PMID- 19135030 TI - Meprin A and meprin alpha generate biologically functional IL-1beta from pro-IL 1beta. AB - The present study demonstrates that both oligomeric metalloendopeptidase meprin A purified from kidney cortex and recombinant meprin alpha are capable of generating biologically active IL-1beta from its precursor pro-IL-1beta. Amino acid sequencing analysis reveals that meprin A and meprin alpha cleave pro-IL 1beta at the His(115)-Asp(116) bond, which is one amino acid N-terminal to the caspase-1 cleavage site and five amino acids C-terminal to the meprin beta site. The biological activity of the pro-IL-1beta cleaved product produced by meprin A, determined by proliferative response of helper T-cells, was 3-fold higher to that of the IL-1beta product produced by meprin beta or caspase-1. In a mouse model of sepsis induced by cecal ligation puncture that results in elevated levels of serum IL-1beta, meprin inhibitor actinonin significantly reduces levels of serum IL-1beta. Meprin A and meprin alpha may therefore play a critical role in the production of active IL-1beta during inflammation and tissue injury. PMID- 19135031 TI - Protective effect against Parkinson's disease-related insults through the activation of XBP1. AB - The accumulation of misfolded and unfolded proteins in endoplasmic reticulum (ER) induces ER stress, activating the unfolded protein response (UPR). Recent evidence has suggested the relationship between UPR and dopaminergic neuronal cell death in Parkinson's disease (PD); however, it remains unclear whether it makes sense to modulate UPR, to mitigate the progression of PD. In this study, we investigated a role of the IRE1 alpha-XBP1 pathway in the survival of dopaminergic cells, under stress induced by PD-related insults. The exogenous expression of the active-form XBP1 (XBP1s) protein had protective effects against cell death induced by 1-methyl-4-phenylpyridinium (MPP+) and proteasome inhibitors. Moreover, adenoviral XBP1s expression significantly suppressed the degeneration of dopaminergic neurons in the mouse model of PD, as induced by 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). These results demonstrate that the enhancement of XBP1 could be a novel PD therapeutic strategy. PMID- 19135032 TI - Microarray expression analysis of human dopaminergic neuroblastoma cells after RNA interference of SNCA--a key player in the pathogenesis of Parkinson's disease. AB - The pre-synaptic protein alpha-synuclein is a key player in the pathogenesis of Parkinson's disease (PD). Misfolded alpha-synuclein protofibrils accumulate and serve as seed structures that cause numerous proteins in the cytoplasm of neuronal cells to aggregate into so-called Lewy bodies. Furthermore, both missense mutations and multiplications of the SNCA gene lead to autosomal dominant forms of familial PD. However, the exact biological role of alpha synuclein in normal brains remains elusive. To gain more insight into the normal function of this protein, we evaluated changes in whole genome expression in dopaminergic neuroblastoma cells (SH-SY5Y) caused by reductions of 90% in alpha synuclein RNA levels and of 59% in alpha-synuclein protein levels as a result of RNA interference. The expression of 361 genes was altered at least+/-1.5-fold by the RNA interference, with 82 up-regulated and 279 down-regulated. The differentially expressed gene products are involved in the regulation of transcription, cell cycle, protein degradation, apoptosis, neurogenesis, and lipid metabolism. To examine the influence of SNCA down-regulation by RNAi on apoptosis, we performed cell death assays using different stress triggers. The changes observed in the expression profile of dopaminergic neuronal cells following reduction of SNCA expression warrant studies to investigate the role of signaling cascades in familial and idiopathic PD. PMID- 19135033 TI - Regulation of Brn-3a N-terminal transcriptional activity by MEK1/2-ERK1/2 signalling in neural differentiation. AB - The POU family transcription factor Brn-3a is required for the differentiation and survival of sensory neurones, and is phosphorylated in neuroblastoma cells following treatment with all-trans retinoic acid (RA). Mutation of serines-121 and -122 of Brn-3a to alanine blocks its phosphorylation and impairs RA-mediated neurite outgrowth. Here we show that this deficit in differentiation is mimicked by a single mutation at serine-122, and demonstrate a similar requirement for a second residue, threonine-39. Like Brn-3a, the neuropeptide Galanin has been implicated in the development of sensory neurones. We show that Brn-3a over expression acts synergistically with RA treatment to up-regulate Galanin promoter activity; that the activity of the N-terminal transcriptional activation domain of Brn-3a is increased following RA treatment; and that both these effects require threonine-39 and serine-122. In addition, we demonstrate that the RA mediated activation of Galanin promoter activity and Brn-3a N-terminal transcriptional activity are both blocked by pan-MEK inhibitors, and show that the expression of a constitutively-active mutant of MEK1, but not MEK5, is sufficient to increase Brn-3a activity. These results reveal an important role for the ERK1/2 pathway in Brn-3a regulation during RA-mediated neuronal differentiation and define the neuropeptide Galanin as a novel target of this transcription factor. PMID- 19135034 TI - ERP correlates of inhibitory and facilitative effects of constituent frequency in compound word reading. AB - In the process of reading compound words, those with high-frequency second constituents are recognized faster than the ones with low-frequency second constituents. However, the role of the first constituent still remains unclear. In the present study, the time course of the frequency effects for both constituents was assessed using Basque compound words embedded in sentences while electrophysiological measures (ERPs) were recorded (Basque is a language with a high frequency of compound words, both right and left-headed). Subjects responded to comprehension questions that were not focused on the compound words. The results revealed that high-frequency first constituents elicited larger negativities starting very early (100-300 ms time window), while low-frequency second constituents elicited larger N400 amplitudes than high-frequency second constituents. Following an activation-verification framework, we argue that the early negativity difference reflects candidate triggering, whereas the N400 difference for the second constituent reflects the cost of its selection and integration for the whole-word meaning to be accessed. PMID- 19135036 TI - Enhanced co-expression of beta-tubulin III and choline acetyltransferase in neurons from mouse embryonic stem cells promoted by icaritin in an estrogen receptor-independent manner. AB - A previous small molecule screen demonstrated that some prenylflavonoids can promote neuronal differentiation from mouse embryonic stem (ES) cells based on morphologic criteria. Here we build on this observation and examine the neuronal subtypes induced by icaritin, a compound screened, and the molecular events underlying the differentiation. In the presence of icaritin, the number of neural rosettes in embryoid bodies (EBs) expressing nestin efficiently increased and the neuroectodermal gene Fgf5 expression upregulated during germ layer formation. The neural progenitors generated from icaritin-treated EBs were further differentiated into the neurons (marked by beta-tubulin III) and also enhanced the choline acetyltransferase (ChAT) expression upon terminal differentiation. A suppression of p38 mitogen-activated protein kinase (p38MAPK) phosphorylation and sustained extracellular signal-regulated protein kinase (ERK) phosphorylation existed simultaneously without estrogen-like activities involved. Taken together, enhanced co-expression of beta-tubulin III and choline acetyltransferase in neuronal differentiation from mouse ES cells is promoted by icaritin via estrogen receptor-independent action. PMID- 19135035 TI - Genetic variance contributes to dopamine receptor antagonist-induced inhibition of sucrose intake in inbred and outbred mouse strains. AB - Preference and intake of sucrose varies across inbred and outbred strains of mice. Pharmacological analyses revealed that the greatest sensitivity to naltrexone-induced inhibition of sucrose (10%) intake was observed in C57BL10/J and C57BL/6J strains, whereas 129P3/J, SWR/J and SJL/J strains displayed far less sensitivity to naltrexone-induced inhibition of sucrose intake. Given that dopamine D1 (SCH23390) and D2 (raclopride) receptor antagonism potently reduce sucrose intake in outbred rat and mouse strains, the present study examined the possibility of genetic variance in the dose-dependent (50-1600 nmol/kg) and time dependent (5-120 min) effects of these antagonists upon sucrose (10%) intake in the eight inbred (BALB/cJ, C3H/HeJ, C57BL/6J, C57BL/10J, DBA/2J, SJL/J, SWR/J and 129P3/J) and one outbred (CD-1) mouse strains previously tested with naltrexone. SCH23390 significantly reduced sucrose intake across all five doses in 129P3/J and SJL/J mice, across four doses in C57BL/6J and BALB/cJ mice, across three doses in DBA/2J, SWR/J, C3H/HeJ and C57BL/10J mice, but only at the two highest doses in CD-1 mice. SCH23390 was 2-3-fold more potent in inhibiting sucrose intake in 129P3/J and SJL/J mice relative to CD-1 mice. In contrast, only the highest equimolar 1600 nmol/kg dose of raclopride significantly reduced sucrose intake in the BALB/cJ, C3H/HeJ, C57BL/6J, C57BL/10J, DBA/2J, SJL/J and 129P3/J, but not the SWR/J and CD-1 strains. The present and previous data demonstrate specific and differential patterns of genetic variability in inhibition of sucrose intake by dopamine and opioid antagonists, suggesting that distinct neurochemical mechanisms control sucrose intake across different mouse strains. PMID- 19135037 TI - In vitro assessment of P450 induction potential of novel chemopreventive agents SR13668, 9-cis-UAB30, and pentamethychromanol in primary cultures of human hepatocytes. AB - Several compounds, including 2,10-dicarbethoxy-6-methoxy-5,7-dihydroindolo[2,3 b]carbazole (SR13668), (2E,4E,6Z,8E)-8-(3',4'-dihydro-1'(2'H)-napthalen-1' ylidene)-3,7-dimethyl-2,4,6-octatrienoic acid (9-cis-UAB30), and 2,2,5,7,8 pentamethyl-6-chromanol (PMCol), were selected as promising chemopreventive agents and have entered preclinical trials for cancer prevention. The potential for adverse drug events resulting from interactions with other administered drugs, food components, or food additives presents an important question. Among the most important drug-drug interactions (DDI) is the potential of a new chemical entity (NCE) to induce cytochrome P450 enzymes (P450). Drug induction of P450 enzymes can lead to adverse drug interactions by increasing the metabolism of other drugs that are substrates for the induced isoform. Currently, sandwich cultured primary human hepatocytes are the standard for predicting human P450 enzyme induction in vitro as these cells retain the ability to respond to prototypical P450 inducers with the same specificity and potency exhibited in vivo. Therefore, a select panel of inducible P450 target genes (CYP1A2, CYP2B6, and CYP3A4) and their induction activity (measured by LC-MS/MS of respective marker substrate metabolites) were monitored in cultured hepatocytes following treatment with SR13668, 9-cis-UAB30, or PMCol to predict clinically significant drug-induced expression. The concentration ranges of the NCE used were selected to maximize the clinical relevance of these results. All responses were evaluated according to major prototypical P450 inducers (i.e., 3-methylcholanthrene, 3-MC; phenobarbital, PB; rifampicin, RIF) and increases > or = 40% of the respective positive control(s) were considered an indication of demonstrable induction. Herein, we report that there is low potential for DDI with SR13668 and PMCol due to enzyme induction of CYP1A2, CYP2B6, and CYP3A4 expression at the concentrations examined. Similarly, the study results suggested that 9-cis-UAB30 has low potential to induce CYP1A2 and CYP3A4 expression at the concentrations examined. However, 9-cis-UAB30 was shown to significantly induce CYP2B6 enzyme activity at 10 microM suggesting the potential for DDI as a result. PMID- 19135038 TI - Interactions of beta-carotene and flavonoids on the secretion of pro-inflammatory mediators in an in vitro system. AB - Chronic inflammation, a process linked to increased oxidative stress, may induce many diseases. Whether beta-carotene prevents inflammation is unclear. Using phorbol-12-myristate-13-acetate (PMA)-stimulated HL-60 cells, we investigated the effects of 2 or 20 microM beta-carotene on the inflammatory reaction of monocyte/macrophage-like cells and the modulation of 20 microM quercetin or naringenin, two flavonoids, of the effects of beta-carotene. The effects of quercetin and naringenin were compared with that of alpha-tocopherol, a well known antioxidant. The stimulated HL-60 cells were also co-incubated with A549 cells to investigate the DNA-damaging ability of the stimulated monocyte/macrophage-like cells on target cells. Our results showed that preincubation with 20 microM beta-carotene significantly enhanced the release of two pro-inflammatory mediators, interleukin-8 and tumor necrosis factor-alpha, in PMA-stimulated HL-60 cells and slightly increased the DNA-damaging ability of these cells. By contrast, 2 microM beta-carotene had an inhibitory effect on the inflammatory reaction in PMA-stimulated cells. The higher dose of beta-carotene also exerted pro-inflammatory effects in lipopolysaccharide-stimulated RAW264.7 cells. Furthermore, quercetin, naringenin, and alpha-tocopherol partly suppressed the pro-inflammatory effects of 20 microM beta-carotene on PMA-stimulated HL-60 cells, and the suppressing effects of quercetin and naringenin were better than or similar to those of alpha-tocopherol. Quercetin also additively or synergistically enhanced the inhibitory effects of 2 microM beta-carotene on the secretion of pro-inflammatory mediators and the DNA-damaging ability of PMA stimulated HL-60 cells. The mechanisms underlying the effect of the flavonoids were associated with their antioxidant activity and inhibition of the production of pro-inflammatory cytokines. Our results urge consideration of the safety of beta-carotene supplementation concerning effects on inflammation and suggest that the interaction between beta-carotene and quercetin or naringenin may alter the effects of beta-carotene on the secretion of pro-inflammatory mediators. PMID- 19135039 TI - Differential effects of silymarin and its active component silibinin on plasma membrane stability and hepatocellular lysis. AB - Silymarin (SIL) is a natural extract with hepatoprotective properties composed mainly of flavonolignans, with silibinin (SB) being its principal constituent. SB is thought to be the main responsible for SIL hepatoprotective properties, although this has not been corroborated systematically. We analysed comparatively the effects of SIL and SB on hepatocellular plasma membrane stability. SIL (500 microM concentration in SB) protected significantly the plasma membrane disruption induced by Triton X-100 (TX-100) and taurochenodeoxycholate (TCDC), both in isolated plasma membrane (assessed by recording the plasma membrane transition from bilayer to micelle using the R18 self-quenching assay) and in isolated rat hepatocytes (assessed by the release into the incubation medium of the cytosolic enzymes lactate dehydrogenase and alanine aminotransferase). Contrarily, SB (500 microM) exacerbated plasma membrane disruption induced by TX 100 in both systems at detergent concentrations relevant to induce hepatocellular lysis, although it displayed some stabilizing properties at higher concentrations. SB showed a lower stabilizing effect against TCDC-induced plasma membrane disruption when assayed in both models. In addition, SB exposure made the plasma membrane more labile to disruption induced by osmotic lysis. These results show that SIL and SB have differential effects on membrane stability; whereas SIL shows consistently stabilizing effects, SB exacerbates hepatocellular lysis or exerts only minimal stabilizing effects. This differential behaviour should be taken into account when considering the pros and cons of using purified SB vs. the whole SIL extract in medicinal formulations for liver diseases. PMID- 19135040 TI - Concentrations of very long-chain fatty acid in whole blood are associated with cardiovascular risk factors in children. AB - BACKGROUND: Fatty acid metabolism has a close relationship with metabolic syndrome. Saturated very long-chain fatty acid (hexacosanoic acid; C26:0) was recently reported to be associated with cardiovascular risk factors in adults. METHODS: Eighty-eight children (47 male, 41 female; average age, 10.9 y) participated in this study. Concentrations of C26:0 in whole blood were measured. RESULTS: Compared with reported concentrations in the whole blood of adults, children had lower C26:0 concentrations, which had a close relationship with abdominal obesity, increased concentrations of low-density lipoprotein cholesterol and high blood pressure. C26:0 concentrations increased with increasing number of risk factors. CONCLUSIONS: Elevation of C26:0 concentrations may be one of the metabolic features of children with cardiovascular risk factors. PMID- 19135041 TI - Interpretation of high sensitivity cardiac troponin I results: reference to biological variability in patients who present to the emergency room with chest pain: case report series. AB - BACKGROUND: The development of highly sensitive cardiac troponin (cTnI) assays has increased the number of true and false positive results for patients suspected of acute myocardial infarction (AMI). Cases are reported whereby the use of serial testing, the 99th percentile cutoff, and the application of biological variation of cTnI were used to help determine ischemic vs. non ischemic causes of myocardial injury. METHODS: cTnI was measured using the Siemens Ultra assay from 13 representative patients who presented to the emergency department with symptoms suggestive of acute cardiac disease. Based on a previous study, reference change values of a 46% increase and 32% decrease were used to interpret results. These differences were compared against the patient's discharge diagnosis. RESULTS: Two patients who subsequently ruled in for AMI had a negative cTnI (<0.04 microg/l) and borderline positive cTnI (0.07 microg/l) at admission, respectively. While the 4-6 h results were also borderline, there was a significant increase from the baseline (+575% and +50%, respectively) to suggest the presence of an acute cardiac event. Two other AMI cases document the significant cTnI decline in results after peak values. In 7 other non-AMI cases (heart and renal failure, gastrointestinal bleeding, stroke and venous thrombosis), while baseline concentrations were clearly positive (0.18-2.12 microg/l), subsequent serial samples were not significantly increased or decreased from baseline. These findings were not typical for AMI. There were 2 cases with acute blunt cardiac trauma and intracranial hemorrhage, respectively, that produced cTnI results that were initially low (<0.04 and 0.05 microg/l, respectively), but significantly increased with serial testing thereby producing false positive Delta cTnI results for AMI. CONCLUSIONS: Serial testing for troponin was useful in differentiating early AMI from non-ischemic causes of troponin increases. However, non-AMI patients with acute cardiac injury can produce troponin results that mimic AMI. Therefore serial troponin testing must be used in conjunction with clinical presentation and other laboratory findings. PMID- 19135042 TI - Cholesterol efflux from J774 macrophages and Fu5AH hepatoma cells to serum is preserved in CETP-deficient patients. AB - BACKGROUND: The role of CETP in the development of atherosclerosis is debatable, and few data exist regarding the total impact of CETP inhibition on cholesterol efflux. METHODS: Acceptor capacities of whole serum and HDL subfractions separated by HPLC were compared using 2 different cell systems. Subjects with CETP deficiency (2 homozygous, 1 compound heterozygous, and 5 heterozygous) were analyzed along with 10 normolipidemic controls. The fractional efflux from cholesterol-labeled Fu5AH hepatoma cells was determined to be SR-BI mediated. The efflux difference between control and liver X receptor (LXR) agonist-induced ABCA1-upregulated J774 macrophages was considered as a measure of ABCA1-mediated efflux. RESULTS: For the Fu5AH cell system, the total acceptor capacities of whole serum and HPLC-separated HDL fraction 2 obtained from the homozygous subjects were 38% and 116% higher than the corresponding values for the controls, respectively (p<0.05). For the J774 cell system, the total acceptor capacities of whole serum and HPLC-separated HDL fractions were similar among the CETP deficient subjects and controls. CONCLUSIONS: Serum from homozygous subjects with CETP-null defects exhibited enhanced acceptor capacity via an SR-BI dependent pathway, which is regulated by the middle HPLC-separated HDL fraction. Further, the cholesterol acceptor capacity of serum obtained from patients having complete and partial CETP deficiency was preserved via an ABCA1-dependent pathway. PMID- 19135043 TI - Analysis of urinary nucleosides as potential tumor markers in human colorectal cancer by high performance liquid chromatography/electrospray ionization tandem mass spectrometry. AB - BACKGROUND: Increased levels of modified nucleosides have been observed in urine from patients suffering from several cancers. In this study, we evaluated whether urinary nucleosides can serve as potential tumor markers for colorectal cancer by high performance liquid chromatography-electrospray/tandem mass spectrometry (HPLC/ESI-MS/MS). METHODS: A simple and specific method based on HPLC/ESI-MS/MS was developed to determine the urinary nucleosides from patients with colorectal cancer. We studied the excretion patterns of nucleosides in urine from 26 patients with colorectal cancer and 18 healthy controls. RESULTS: The LC/MS/MS approach with selective reaction monitoring (SRM) allowed for the sensitive determination of nucleosides in human urine samples with colorectal cancer. The mean levels of 5 urinary nucleosides (adenosine, cytidine, N(2),N(2) dimethylguanine, 8-hydroxy-2'-deoxyguanosine and uridine) were significantly higher in the patients with colorectal cancer than in the healthy adults. CONCLUSIONS: The results indicate that urinary nucleosides determined by LC/MS/MS may be useful as biological markers for colorectal cancer. Our findings suggest that LC/MS/MS is a highly specific and sensitive method for rapidly screening a large number of nucleoside that may be useful as markers for cancer in humans. PMID- 19135044 TI - Two-round allele specific-polymerase chain reaction: a simple and highly sensitive method for JAK2V617F mutation detection. AB - BACKGROUND: Philadelphia chromosome (Ph(1))-negative myeloproliferative disorders (MPD) are hematopoietic stem cell disorders characterized by extensive proliferation of myeloid blood cells. JAK2V617F has recently been identified in the majority of Ph(1)-negative MPD and becomes an essential diagnostic marker. METHODS: To screen for JAK2V617F, a two-round allele specific-polymerase chain reaction (AS-PCR) was developed and compared to PCR-restriction fragment length polymorphism (PCR-RFLP), denaturing high performance liquid chromatography (DHPLC), and DNA sequencing. A primary AS-PCR was performed followed by a secondary AS-PCR, which was an amplification of the primary AS-PCR products using the same set of primers under alternative conditions. RESULTS: By primary AS-PCR, a strong mutant-DNA band was seen in the DNA mixture containing as low as 2.5% of mutant allele. An ambiguous band was seen in 1% dilution while being totally absent in 0.1% dilution. After secondary AS-PCR, a mutant DNA band was clearly detected at 0.01% dilution. The detection sensitivity of PCR-RFLP and DHPLC was 2.5% while sequencing analysis was unable to detect below 5% dilution. CONCLUSION: Two-round AS-PCR is simple and inexpensive, making it a suitable method for JAK2V617F mutation screening. Moreover, monitoring of minimal residual disease after specific treatment of Ph(1)-negative MPD patients should be feasible with this highly sensitive method. PMID- 19135045 TI - Retinal determination genes as targets and possible effectors of extracellular signals. AB - Retinal determination genes are sufficient to specify eyes in ectopic locations, raising the question of how these master regulatory genes define an eye developmental field. Genetic mosaic studies establish that expression of the retinal determination genes eyeless, teashirt, homothorax, eyes absent, sine oculis, and dachshund are each regulated by combinations of Dpp, Hh, N, Wg, and Ras signals in Drosophila. Dpp and Hh control eyeless, teashirt, sine oculis, and dachshund expression, Dpp and Ras control homothorax, and all the signaling pathways affect eyes absent expression. These results suggest that eye-specific development uses retinal determination gene expression to relay positional information to eye target genes, because the distinct, overlapping patterns of retinal determination gene expression reflect the activities of the extracellular signaling pathways. PMID- 19135046 TI - Increased thymus- and decreased parathyroid-fated organ domains in Splotch mutant embryos. AB - Embryos that are homozygous for Splotch, a null allele of Pax3, have a severe neural crest cell (NCC) deficiency that generates a complex phenotype including spina bifida, exencephaly and cardiac outflow tract abnormalities. Contrary to the widely held perception that thymus aplasia or hypoplasia is a characteristic feature of Pax3(Sp/Sp) embryos, we find that thymic rudiments are larger and parathyroid rudiments are smaller in E11.5-12.5 Pax3(Sp/Sp) compared to Pax3(+/+) embryos. The thymus originates from bilateral third pharyngeal pouch primordia containing endodermal progenitors of both thymus and parathyroid glands. Analyses of Foxn1 and Gcm2 expression revealed a dorsal shift in the border between parathyroid- and thymus-fated domains at E11.5, with no change in the overall cellularity or volume of each shared primordium. The border shift increases the allocation of third pouch progenitors to the thymus domain and correspondingly decreases allocation to the parathyroid domain. Initial patterning in the E10.5 pouch was normal suggesting that the observed change in the location of the organ domain interface arises during border refinement between E10.5 and E11.5. Given the well-characterized NCC defects in Splotch mutants, these findings implicate NCCs in regulating patterning of third pouch endoderm into thymus- versus parathyroid-specified domains, and suggest that organ size is determined in part by the number of progenitor cells specified to a given fate. PMID- 19135047 TI - Effects of systemic 5-HT(1B) receptor compounds on ventral tegmental area intracranial self-stimulation thresholds in rats. AB - Serotonin 1B (5-HT(1B)) receptors may play a role in regulating motivation and reward-related behaviours. To date, no studies have investigated the effects of the highly selective 5-HT(1B) receptor agonist CP 94253, on the reward model of ventral tegmental area intracranial self-stimulation. The current study investigated the hypothesis that 5-HT(1B) receptors play an inhibitory role in ventral tegmental area ICSS. Using Sprague-Dawley rats, the effects of the selective 5-HT(1B) receptor agonist CP 94253 (0-5.0 mg/kg) and the 5-HT(1B/1D) receptor antagonist GR 127935 (10.0 mg/kg) were investigated in rats trained to respond for ventral tegmental area ICSS; results were compared using rate frequency threshold analysis. The highest dose of CP 94253 (5.0 mg/kg) tested in ventral tegmental area ICSS produced an increase in rate-frequency thresholds without affecting maximal response rates. This effect was attenuated by GR 127935 which did not show any effects when administered alone. These results suggest that 5-HT(1B) receptors play an inhibitory role in regulating ventral tegmental area ICSS. PMID- 19135048 TI - Oxaloacetate restores the long-term potentiation impaired in rat hippocampus CA1 region by 2-vessel occlusion. AB - Various acute brain pathological conditions are characterized by the presence of elevated glutamate concentrations in the brain interstitial fluids. It has been established that a decrease in the blood glutamate level enhances the brain-to blood efflux of glutamate, removal of which from the brain may prevent glutamate excitotoxicity and its contribution to the long-lasting neurological deficits seen in stroke. A decrease in blood glutamate level can be achieved by exploiting the glutamate-scavenging properties of the blood-resident enzyme glutamate oxaloacetate transaminase, which transforms glutamate into 2-ketoglutarate in the presence of the glutamate co-substrate oxaloacetate. The present study had the aim of an evaluation of the effects of the blood glutamate scavenger oxaloacetate on the impaired long-term potentiation (LTP) induced in the 2-vessel occlusion ischaemic model in rat. Transient (30-min) incomplete forebrain ischaemia was produced 72 h before LTP induction. Although the short transient brain hypoperfusion did not induce histologically identifiable injuries in the CA1 region (Fluoro-Jade B, S-100 and cresyl violet), it resulted in an impaired LTP function in the hippocampal CA1 region without damaging the basal synaptic transmission between the Schaffer collaterals and the pyramidal neurons. This impairment could be fended off in a dose-dependent manner by the intravenous administration of oxaloacetate in saline (at doses between 1.5 mmol and 0.1 mumol) immediately after the transient hypoperfusion. Our results suggest that oxaloacetate-mediated blood and brain glutamate scavenging contributes to the restoration of the LTP after its impairment by brain ischaemia. PMID- 19135049 TI - Antidepressants reveal differential effect against 1-methyl-4-phenylpyridinium toxicity in differentiated PC12 cells. AB - Treatment of depression may ameliorate the cognitive disability and motor slowness in Parkinson's disease. It has been shown that antidepressants, including fluoxetine, may attenuate or exacerbate neuronal cell death. The present study assessed the effect of antidepressants (amitriptyline, tranylcypromine and fluoxetine) against the toxicity of 1-methyl-4 phenylpyridinium (MPP(+)) in relation to the mitochondria-mediated cell death process in differentiated PC12 cells. Amitriptyline and tranylcypromine attenuated the MPP(+)-induced cell death that may be associated with mitochondrial membrane permeability change and oxidative stress. Both compounds prevented the loss of the mitochondrial transmembrane potential, over-expression of Bax, reduction in Bcl-2 level, cytochrome c release, caspase-3 activation, formation of reactive oxygen species and depletion of GSH. The inhibitory effect of tranylcypromine was greater than that of amitriptyline on the basis of concentration. In contrast, fluoxetine revealed a toxic effect and exhibited an additive effect against the toxicity of MPP(+). Results show that amitriptyline and tranylcypromine may attenuate the MPP(+) toxicity by suppressing the mitochondrial membrane permeability change that leads to cytochrome c release and subsequent caspase-3 activation. The effects seem to be associated with the inhibitory action on the formation of reactive oxygen species and the depletion of GSH. In contrast, fluoxetine seems to exert an additive toxic effect against neuronal cell damage by increasing mitochondrial damage and oxidative stress. PMID- 19135050 TI - Mechanism for resveratrol-induced cardioprotection against reperfusion injury involves glycogen synthase kinase 3beta and mitochondrial permeability transition pore. AB - Resveratrol pretreatment can protect the heart by inducing pharmacological preconditioning. Whether resveratrol protects the heart when applied at reperfusion remains unknown. We examined the effect of resveratrol on myocardial infarct size when given at reperfusion and investigated the mechanism underlying the effect. Isolated rat hearts were subjected to 30 min ischemia followed by 2 h of reperfusion, and myocardial samples were collected from the risk zone for Western blot analysis. Mitochondrial swelling was spectrophotometrically measured as a decrease in absorbance at 520 nm (A(520)). Resveratrol reduced infarct size and prevented cardiac mitochondrial swelling. Resveratrol enhanced GSK-3beta phosphorylation upon reperfusion, an effect that was mediated by the cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG) pathway. Resveratrol translocated GSK-3beta from cytosol to mitochondria via the cGMP/PKG pathway. Further studies showed that mitochondrial GSK-3beta was co-immunoprecipitated with cyclophilin D but not with VDAC (voltage dependent anion channel) or ANT (adenine nucleotide translocator). These data suggest that resveratrol prevents myocardial reperfusion injury presumably by targeting the mPTP through translocation of GSK-3beta from cytosol to mitochondria. Translocated GSK-3beta may ultimately interact with cyclophilin D to modulate the mPTP opening. PMID- 19135051 TI - Expression of ALK protein, mRNA and fusion transcripts in anaplastic large cell lymphoma. AB - Systemic anaplastic large cell lymphoma (ALCL) can be divided into two subgroups, anaplastic lymphoma kinase (ALK)-positive and ALK-negative, based on the expression of ALK protein. Expression of this protein is due to genetic alterations of ALK at 2p23. Overall, observations on ALK protein, ALK mRNA, ALK associated genetic alterations and their relationships, to one another are not often reported in the literature. In this study, we investigated the expression of ALK protein, mRNA and fusion transcripts involving ALK and their relationships in ALCL and analyzed formalin-fixed, paraffin-embedded tissues. Forty-five human cases were analyzed with immunohistochemistry for the ALK protein and RT-PCR for ALK mRNA and seven kinds of ALK involved fusion transcripts. Our results showed that the expression of ALK protein, ALK mRNA and ALK fusion transcripts were significantly related to one another (P<0.01). Consistent with the expression of ALK protein, patients presenting with ALK mRNA or ALK involved fusion transcripts were significantly younger than those lacking ALK gene alteration (P<0.01). This study demonstrates expression of both ALK protein and ALK mRNA are positively correlated with expression of ALK-associated fusion transcripts. Combined detection of ALK protein, ALK mRNA and ALK fusion transcripts can complement each other to aid in the diagnosis of ALCL. PMID- 19135052 TI - Protein aging: truncation of aquaporin 0 in human lens regions is a continuous age-dependent process. AB - The human lens is ideal for the study of macromolecular aging because cells in the centre, along with their constituent proteins, are present for our entire lives. We examined the major membrane protein, aquaporin 0 (AQP0), in regions of the lens formed at different times during our lifespan, to determine if similar changes could be detected and if they were progressive. Membrane fractions from three concentric lens regions were examined by SDS-PAGE coupled with densitometry, and Western blotting, to assess the time course of truncation. The overall extent of modification was also examined by MALDI mass spectrometry of the undigested proteins. In all regions, AQP0 became progressively more truncated, specifically by the loss of a 2kDa intracellular C-terminal peptide. The proteolysis increased steadily in all regions such that half of the AQP0 in the barrier region (that part of the lens formed immediately after birth) had been cleaved by age 40-50. MALDI mass spectrometry revealed that in all regions, AQP0 not only was shortened, it also became progressively more heterogeneous with age. Since the lens interior is devoid of active enzymes, it is very likely that the cleavage of AQP0 is chemically induced. We speculate that the loss of this C terminal peptide 'spacer' may allow occlusion of AQP0 pores on the cytoplasmic face of the fibre cell membranes. Once a significant proportion of AQP0 has been cleaved, this occlusion may contribute to the formation of the lens permeability barrier that develops at middle age. PMID- 19135054 TI - Peptidergic neurosecretory cells in insects: organization and control by the bHLH protein DIMMED. AB - This review considers evidence that defines a role for the transcription factor DIMMED in the regulation of insect neurosecretory cells. Genetic anatomical and molecular data all suggest DIMMED is a dedicated controller of the regulated secretory pathway. DIMM is normally expressed within diverse neuropeptide expressing cells and appears highly correlated with a neurosecretory cell fate. Loss of DIMM is associated with deficits in display of neuropeptides and neuropeptide-associated enzymes. Gain of DIMM promotes such display in peptidergic cells and can confer such neurosecretory properties onto conventional neurons. We review models proposed to explain how DIMMED regulates these essential cellular properties. PMID- 19135053 TI - Leishmania (Viannia) braziliensis genotypes identified in lesions of patients with atypical or typical manifestations of tegumentary leishmaniasis: evaluation by two molecular markers. AB - Analyses of MLEE, RAPD and LSSP-PCR were used to compare the panel of american tegumentary leishmaniasis (ATL) isolates obtained from lesions of patients with rare clinical manifestations of the disease and typical lesions. All of the 34 samples analyzed by MLEE demonstrated similar electromorphic profiles with Leishmania (Viannia) braziliensis reference strain. Through the RAPD analysis, nine genetic profiles (genotypes) were identified. LSSP-PCR corroborates the initial screening and phenetic analysis has grouped the isolates into two major clusters comprising the nine different genotypes. Prevalent genotype defined as LbmtDNAgen1 was detected in the largest number of isolates. There was no association between genotypes and clinical symptoms. However, two different genotypes could be identified in the initial (LbmtDNAGen9) and reactivated lesion (LbmtDNAGen3) of the same patient. Our results support the idea of a less pronounced genotypic diversity among L. (V.) braziliensis circulating in the State of Rio de Janeiro and demonstrate the useful application of these molecular markers in genetics variability studies. PMID- 19135055 TI - Neuropeptidases and the metabolic inactivation of insect neuropeptides. AB - Neuropeptidases play a key role in regulating neuropeptide signalling activity in the central nervous system of animals. They are oligopeptidases that are generally found on the surface of neuronal cells facing the synaptic and peri synaptic space and therefore are ideally placed for the metabolic inactivation of neuropeptide transmitters/modulators. This review discusses the structure of insect neuropeptides in relation to their susceptibility to hydrolysis by peptidases and the need for specialist enzymes to degrade many neuropeptides. It focuses on five neuropeptidase families (neprilysin, dipeptidyl-peptidase IV, angiotensin-converting enzyme, aminopeptidase and dipeptidyl aminopeptidase III) that have been implicated in the metabolic inactivation of neuropeptides in the central nervous system of insects. Experimental evidence for the involvement of these peptidases in neuropeptide metabolism is reviewed and their properties are compared to similar neuropeptide inactivating peptidases of the mammalian brain. We also discuss how the sequencing of insect genomes has led to the molecular identification of candidate neuropeptidase genes. PMID- 19135056 TI - The cytosolic tryparedoxin of Leishmania infantum is essential for parasite survival. AB - Leishmania infantum cytosolic tryparedoxin (LiTXN1) can be regarded as a potential candidate for drug targeting. This redox active molecule, which belongs to the thioredoxin superfamily, is one constituent of the hydroperoxide elimination cascade in L. infantum and may also be involved in other cellular processes such as DNA synthesis or host-parasite interaction. In order to validate LiTXN1 as a drug target we have employed a gene replacement strategy. We observed that substitution of both chromosomal LiTXN1 alleles was only possible upon parasite complementation with an episomal copy of the gene. Furthermore, contrary to control parasites carrying the empty vector, both the insect and the mammalian stages of L. infantum retained the episomal copy of LiTXN1 in the absence of drug pressure. These results confirm the essentiality of LiTXN1 throughout the life cycle of the parasite, namely in the disease-causing amastigote stage. In addition, the data obtained showed that disruption of one allele of this gene leads only to a 25% reduction in the expression of LiTXN1. Even though this does not affect promastigote growth and susceptibility to hydrogen peroxide, ex vivo infection assays suggest that wild-type levels of LiTXN1 are required for optimal L. infantum virulence. PMID- 19135057 TI - Silencing of acetylcholinesterase gene of Helicoverpa armigera by siRNA affects larval growth and its life cycle. AB - RNA interference is an effective means of regulation of gene expression both in vitro and in vivo. We studied the effect of siRNA on larval development by selective targeting of the acetylcholinesterase (AChE) gene of Helicoverpa armigera. Chemically synthesized siRNA molecules were directly fed to H. armigera larvae along with the artificial diet. The siRNA treatment resulted in specific gene silencing of AChE and consequently brought about mortality, growth inhibition of larvae, reduction in the pupal weight, malformation and drastically reduced fecundity as compared to control larvae. Our studies suggest some novel roles for AChE in growth and development of insect larvae and demonstrate that siRNA can be readily taken up by insect larvae with their diet. PMID- 19135058 TI - Impact of transgenic oilseed rape expressing oryzacystatin-1 (OC-1) and of insecticidal proteins on longevity and digestive enzymes of the solitary bee Osmia bicornis. AB - The risk that insect-resistant transgenic plants may pose for solitary bees was assessed by determining longevity of adult Osmia bicornis (O. rufa) chronically exposed to transgenic oilseed rape expressing oryzacystatin-1 (OC-1) or to the purified insecticidal proteins recombinant rOC-1, Kunitz soybean trypsin inhibitor (SBTI), Galanthus nivalis agglutinin (GNA), or Bacillus thuringiensis toxin Cry1Ab dissolved in sugar solution (at 0.01 and 0.1%, w:v, Cry1Ab only at 0.01%). Compared to control bees, longevity was significantly reduced by SBTI and GNA at both concentrations and by rOC-1 at 0.1%, but not by Cry1Ab or rOC-1 at 0.01%. Longevity on the OC-1 oilseed rape was not significantly different from the control plants. The effects of SBTI and rOC-1 on longevity were investigated through characterization of the digestive proteinases of O. bicornis and analysis of the response in proteinase profiles to ingestion of these proteinase inhibitors. A relatively complex profile of at least four types of soluble proteolytic enzymes was identified. Serine proteinases were found to be predominant, with metallo and especially cysteine proteinases making a smaller albeit significant contribution. The compensatory response to in vivo enzyme inhibition was similar for SBTI and rOC-1 although less pronounced for rOC-1. It consisted of a non-specific overproduction of native proteinases, both sensitive and insensitive, and the induction of a novel aspartic proteinase. PMID- 19135059 TI - Differential control of cell death and gene expression during two distinct phases of hormonally-regulated muscle death in the tobacco hawkmoth Manduca sexta. AB - In larvae of the tobacco hawkmoth Manduca sexta, the intersegmental muscles (ISMs) span eight abdominal segments and represent the major muscle group. Following pupation, the ISMs in the first two and last two segments undergo programmed cell death (PCD), while the remaining four segments persist until the time of adult eclosion, when they too undergo PCD. ISM death at adult eclosion is initiated by a decline in the circulating ecdysteroid titer and requires de novo gene expression. In this study we have investigated the hormonal regulation and the patterns of gene expression that accompany both early and late ISM death. We find that distinct endocrine cues regulate these two periods of muscle death. Even though the middle segments of ISMs are exposed to the same endocrine environment as the adjacent cells that die following pupation, they do not express death-associated transcripts until they are specifically signaled to die following adult eclosion. These data indicate that subsets of homologous muscles appear to make segment-specific decisions to couple their endogenous cell death programs to distinctly different developmental cues. Nevertheless, once cell death is initiated, they utilize many of the same molecular components. PMID- 19135060 TI - Molecular composition and functional properties of f-channels in murine embryonic stem cell-derived pacemaker cells. AB - Mouse embryonic stem cells (mESCs) differentiate into all cardiac phenotypes, and thus represent an important potential source for cardiac regenerative therapies. Here we characterize the molecular composition and functional properties of "funny" (f-) channels in mESC-derived pacemaker cells. Following differentiation, a fraction of mESC-derived myocytes exhibited action potentials characterized by a slow diastolic depolarization and expressed the I(f) current. I(f) plays an important role in the pacemaking mechanism of these cells since ivabradine (3 microM), a specific f-channel inhibitor, inhibited I(f) by about 50% and slowed rate by about 25%. Analysis of I(f) kinetics revealed the presence of two populations of cells, one expressing a fast- and one a slow-activating I(f); the two components are present both at early and late stages of differentiation and had also distinct activation curves. Immunofluorescence analysis revealed that HCN1 and HCN4 are the only isoforms of the pacemaker channel expressed in these cells. Rhythmic cells responded to beta-adrenergic and muscarinic agonists: isoproterenol (1 microM) accelerated and acetylcholine (0.1 microM) slowed spontaneous rate by about 50 and 12%, respectively. The same agonists caused quantitatively different effects on I(f): isoproterenol shifted activation curves by about 5.9 and 2.7 mV and acetylcholine by -4.0 and -2.0 mV in slow and fast I(f)-activating cells, respectively. Accordingly, beta1- and beta2-adrenergic, and M2-muscarinic receptors were detected in mESC-derived myocytes. Our data show that mESC-derived pacemaker cells functionally express proteins which underlie generation and modulation of heart rhythm, and can therefore represent a potential cell substrate for the generation of biological pacemakers. PMID- 19135061 TI - A mathematical model of alpha-catenin dimerization at adherens junctions in polarized epithelial cells. AB - The protein alpha-catenin is found as a monomer or homodimer. As a monomer, alpha catenin can bind to beta-catenin, which localizes to the plasma membrane at the site of adherens junctions (AJs) in polarized epithelial cells. As a dimer, alpha catenin can bind to actin filaments, affecting the organization of the actin cytoskeleton. At usual cytoplasmic concentrations, alpha-catenin is found predominantly in monomeric form. It is currently thought that alpha-catenin cannot simultaneously bind beta-catenin and homodimerize, and that the dynamics of binding and unbinding from beta-catenin, possibly coupled with lower diffusion near an AJ, are sufficient to locally accumulate alpha-catenin monomers and homodimers. Using a mathematical model of alpha-catenin dynamics, I show that alpha-catenin must transiently homodimerize while bound to beta-catenin in order for homodimers to form, even in the presence of a spatial diffusion gradient. PMID- 19135062 TI - Voting with your feet: Payoff biased migration and the evolution of group beneficial behavior. AB - Human migration is nonrandom. In small scale societies of the past, and in the modern world, people tend to move to wealthier, safer, and more just societies from poorer, more violent, less just societies. If immigrants are assimilated, such nonrandom migration can increase the occurrence of culturally transmitted beliefs, values, and institutions that cause societies to be attractive to immigrants. Here we describe and analyze a simple model of this process. This model suggests that long run outcomes depend on the relative strength of migration and local adaptation. When local adaption is strong enough to preserve cultural variation among groups, cultural variants that make societies attractive always predominate, but never drive alternative variants to extinction. When migration predominates, outcomes depend both on the relative attractiveness of alternative variants and on the initial sizes of societies that provide and receive immigrants. PMID- 19135063 TI - A model of the complex response of Staphylococcus aureus to methicillin. AB - It is widely accepted that beta-lactam antimicrobials cause cell death through a mechanism that interferes with cell wall synthesis. Later studies have also revealed that beta-lactams modify the autolysis function (the natural process of self-exfoliation of the cell wall) of cells. The dynamic equilibrium between growth and autolysis is perturbed by the presence of the antimicrobial. Studies with Staphylococcus aureus to determine the minimum inhibitory concentration (MIC) have revealed complex responses to methicillin exposure. The organism exhibits four qualitatively different responses: homogeneous sensitivity, homogeneous resistance, heterogeneous resistance and the so-called 'Eagle effect'. A mathematical model is presented that links antimicrobial action on the molecular level with the overall response of the cell population to antimicrobial exposure. The cell population is modeled as a probability density function F(x,t) that depends on cell wall thickness x and time t. The function F(x,t) is the solution to a Fokker-Planck equation. The fixed point solutions are perturbed by the antimicrobial load and the advection of F(x,t) depends on the rates of cell wall synthesis, autolysis and the antimicrobial concentration. Solutions of the Fokker-Planck model are presented for all four qualitative responses of S. aureus to methicillin exposure. PMID- 19135064 TI - Emergence and loss of assortative mating in sympatric speciation. AB - We have studied an agent model which presents the emergence of sexual barriers through the onset of assortative mating, a condition that might lead to sympatric speciation. In the model, individuals are characterized by two traits, each determined by a single locus A or B. Heterozygotes on A are penalized by introducing an adaptive difference from homozygotes. Two niches are available. Each A homozygote is adapted to one of the niches. The second trait, called the marker trait has no bearing on the fitness. The model includes mating preferences, which are inherited from the mother and subject to random variations. A parameter controlling recombination probabilities of the two loci is also introduced. We study the phase diagram by means of simulations, in the space of parameters (adaptive difference, carrying capacity, recombination probability). Three phases are found, characterized by (i) assortative mating, (ii) extinction of one of the A alleles and (iii) Hardy-Weinberg like equilibrium. We also make perturbations of these phases to see how robust they are. Assortative mating can be gained or lost with changes that present hysteresis loops, showing the resulting equilibrium to have partial memory of the initial state and that the process of going from a polymorphic panmictic phase to a phase where assortative mating acts as sexual barrier can be described as a first-order transition. PMID- 19135065 TI - Modelling chemotherapy resistance in palliation and failed cure. AB - The goal of palliative cancer chemotherapy treatment is to prolong survival and improve quality of life when tumour eradication is not feasible. Chemotherapy protocol design is considered in this context using a simple, robust, model of advanced tumour growth with Gompertzian dynamics, taking into account the effects of drug resistance. It is predicted that reduced chemotherapy protocols can readily lead to improved survival times due to the effects of competition between resistant and sensitive tumour cells. Very early palliation is also predicted to quickly yield near total tumour resistance and thus decrease survival duration. Finally, our simulations indicate that failed curative attempts using dose densification, a common protocol escalation strategy, can reduce survival times. PMID- 19135066 TI - Beneficial effects of intercellular interactions between pancreatic islet cells in blood glucose regulation. AB - Glucose homeostasis is controlled by the islets of Langerhans which are equipped with alpha-cells increasing the blood glucose level, beta-cells decreasing it, and delta-cells the precise role of which still needs identifying. Although intercellular communications between these endocrine cells have recently been observed, their roles in glucose homeostasis have not been clearly understood. In this study, we construct a mathematical model for an islet consisting of two state alpha-, beta-, and delta-cells, and analyze effects of known chemical interactions between them with emphasis on the combined effects of those interactions. In particular, such features as paracrine signals of neighboring cells and cell-to-cell variations in response to external glucose concentrations as well as glucose dynamics, depending on insulin and glucagon hormone, are considered explicitly. Our model predicts three possible benefits of the cell-to cell interactions: First, the asymmetric interaction between alpha- and beta cells contributes to the dynamic stability while the perturbed glucose level recovers to the normal level. Second, the inhibitory interactions of delta-cells for glucagon and insulin secretion prevent the wasteful co-secretion of them at the normal glucose level. Finally, the glucose dose-responses of insulin secretion is modified to become more pronounced at high glucose levels due to the inhibition by delta-cells. It is thus concluded that the intercellular communications in islets of Langerhans should contribute to the effective control of glucose homeostasis. PMID- 19135067 TI - Spiders as biological controllers in the agroecosystem. AB - In this paper, we propose a general model consisting of insects, pests and spiders interacting in an agroecosystem included in a typical homogeneous rural landscape, characterized by a continuous mosaic of cultivated land and a few small patches of grasslands and small woods bounding the fields. The model is general enough to show all the phenomena observed in the agroecosystem. The role of the spider population as a biological controller in the agroecosystem is particularly emphasized. Human intervention by means of pesticide spraying and its relationship with the biological pest controllers is also accounted for. PMID- 19135068 TI - A note on a method for determining advantageous properties of an anti-arrhythmic drug based on a mathematical model of cardiac cells. AB - Regional hyperkalemia during acute ischemia may provoke cardiac arrhythmias such as ventricular fibrillation. Despite intense research efforts over the last decades, the problem of finding an efficient anti-arrhythmic drug without dangerous side effects is still open. One approach to analyze the effect of anti arrhythmic drugs is to do simulations based on mathematical models of collections of cardiomyocytes. Such simulations have recently illuminated the pro-arrhythmic capability of well-established anti-arrhythmic drugs. The purpose of the present note is to introduce a method intended for computing advantageous properties of an anti-arrhythmic drug. For a given model of a normal and an ischemic cell, we introduce a drug as a vector of non-negative real numbers whose components are multiplied by individual terms representing specific ionic currents. The drug vector is computed such that the action potentials of the resulting drugged cells are as close as possible to the action potential of a normal (not drugged) cell. Numerical simulations based on the Luo-Rudy I model and the Hund-Rudy model show that the classical shortened action potential obtained due to hyperkalemia is prolonged by using the drug computed by this method. Furthermore, for both models a 2D collection of spatially coupled ischemic cells give arrhythmogenic solutions before the drug is applied, and stable solutions after the drug is applied. It is emphasized that we do not address the possibility of realizing a drug with the properties computed in this note. PMID- 19135069 TI - Spaceflight induces both transient and heritable alterations in DNA methylation and gene expression in rice (Oryza sativa L.). AB - Spaceflight represents a complex environmental condition in which several interacting factors such as cosmic radiation, microgravity and space magnetic fields are involved, which may provoke stress responses and jeopardize genome integrity. Given the inherent property of epigenetic modifications to respond to intrinsic as well as external perturbations, it is conceivable that epigenetic markers like DNA methylation may undergo alterations in response to spaceflight. We report here that extensive alteration in both DNA methylation and gene expression occurred in rice plants subjected to a spaceflight, as revealed by a set of characterized sequences including 6 transposable elements (TEs) and 11 cellular genes. We found that several features characterize the alterations: (1) All detected alterations are hypermethylation events; (2) whereas alteration in both CG and CNG methylation occurred in the TEs, only alteration in CNG methylation occurred in the cellular genes; (3) alteration in expression includes both up- and down-regulations, which did not show a general correlation with alteration in methylation; (4) altered methylation patterns in both TEs and cellular genes are heritable to progenies at variable frequencies; however, stochastic reversion to wild-type patterns and further de novo changes in progenies are also apparent; and (5) the altered expression states in both TEs and cellular genes are also heritable to selfed progenies but with markedly lower transmission frequencies than altered DNA methylation states. Furthermore, we found that a set of genes encoding for the various putative DNA methyltransferases, 5-methylcytosine DNA glycosylases, the SWI/SNF chromatin remodeller (DDM1) and siRNA-related proteins are extremely sensitive to perturbation by spaceflight, which might be an underlying cause for the altered methylation patterns in the space-flown plants. We discuss implications of spaceflight-induced epigenetic variations with regard to health safety issues of spaceship crews and potentiality of spaceflight as a means for mutagenesis in crop breeding. PMID- 19135070 TI - Nitric oxide utilizes NF-kappaB to signal its neuroprotective effect against alcohol toxicity. AB - Alcohol damages the developing brain and can lead to fetal alcohol syndrome. One of alcohol's most important neuropathologic effects is neuronal death. As neurons mature, they become less vulnerable to alcohol-induced death because they acquire a protective signaling pathway, mediated by nitric oxide (NO). This pathway is the NO-cGMP-cyclic GMP-dependent protein kinase G (NO-cGMP-PKG) pathway. The goal of the present studies was to determine whether nuclear factor kappa B (NF kappaB) is the downstream effector through which the NO-cGMP-PKG pathway signals its neuroprotective effects against alcohol. An activator of NF-kappaB, tumor necrosis factor-alpha (TNF-alpha), protected immature cerebellar granule neuron cultures against alcohol-induced cell death in a dose-dependent fashion. The protective effect of TNF-alpha was similar in magnitude to the protective effects of NMDA and DETA-NONOate, both of which are NO-cGMP-PKG pathway activators. Blockade of the pathway at its first step with NAME, second step with LY83583, or third step with PKG inhibitor increased alcohol-induced cell death and the vulnerability of mature neurons to alcohol toxicity. TNF-alpha protected the neurons, even when the NO-cGMP-PKG pathway was blocked at upstream sites. NF kappaB activation inhibitor (NFi) worsened alcohol-induced cell death and blocked the protective effects of NO-cGMP-PKG pathway activators and TNF-alpha. TNF-alpha reduced the alcohol vulnerability of immature neurons, while NFi increased the vulnerability of mature neurons. Both NMDA and TNF-alpha led to the phosphorylation and degradation of IkappaBalpha, demonstrating that both agents can activate NF-kappaB in cerebellar granule cells. Thus, NF-kappaB plays a critical role in the acquisition of alcohol resistance by maturing neurons and is a key downstream effector through which the NO-cGMP-PKG pathway signals its neuroprotective effects against alcohol. PMID- 19135071 TI - In the eye of the beholder: individual differences in reward-drive modulate early frontocentral ERPs to angry faces. AB - Individual differences in reward-drive have been associated with increased attention toward facial signals of aggression, heightened experience of anger and vulnerability to display aggressive behaviour. Recent fMRI research suggests that these effects rely on reduced ventromedial prefrontal (and increased amygdala) response to aggressive facial displays compared with neutral and sad expressions in subjects scoring high on reward-drive. However, nothing is known about the timing of this modulation. Using event-related potentials (ERPs), we provide the first evidence that greater proneness to display hostile and aggressive behaviour (measured by high scores on the reward-drive) is associated with a reduced midline frontocentral response to aggressive faces within 200-300ms. In addition to confirming a particular interaction between anger processing and aggression related personality traits in ventromedial prefrontal brain regions, our study brings a first indication of when their interaction occurs in the brain, strengthening results from previous classical as well as functional connectivity fMRI studies. PMID- 19135072 TI - Perspective-taking as part of narrative comprehension: a functional MRI study. AB - During narrative comprehension, readers understand the emotions of the protagonist by taking the perspective of the character, which is an essential component of empathy. Spatial perspective-taking is crucial to understanding the standpoints and perceptions of others, and gives clues as to what the protagonist knows. As a default, a "here and now" point-of-view is adopted to make sense of the narrative. If the protagonist is in a different location while an event takes place ("there and now"), in order to comprehend the narrative the reader must take an allocentric perspective, which places greater demands on spatial perspective-taking. Utilizing this phenomenon, we evaluated the neural substrates of perspective-taking in emotional narrative comprehension using functional MRI in 18 normal adults. The subjects read short stories followed by a target sentence, which described an event that might evoke an emotional response in the protagonist if the character were present. The stories involved a scenario in which the character was either present at the same location ("here and now") or at a distant location ("there and now") during the event. The "there and now" scenario activated the posterior cingulate cortex and the right temporo-parietal junction more prominently than the "here and now" condition. In contrast to the control tasks, both scenarios activated the well-known mentalizing network including the dorsomedial prefrontal cortex, temporal pole, posterior cingulate cortex and temporo-parietal junction. Along with the mentalizing network, the posterior cingulate cortex and the right temporo-parietal junction are involved in spatial perspective-taking during emotional narrative comprehension. PMID- 19135073 TI - The volatility of an alarm pheromone in male rats. AB - The volatility of an alarm pheromone in male rats. PHYSIOL BEHAV 00(0) 000-000, 2008. We previously reported that an alarm pheromone released from the perianal region of male rats is perceived by the vomeronasal organ and evokes stress induced hyperthermia and defensive and risk assessment behavior. In addition, we recently reported that the alarm pheromone enhances the acoustic startle reflex (ASR). However, in contrast to our knowledge about such biological aspects of the pheromone, information concerning the physical character of the alarm pheromone is extremely limited. In this study, we investigated the volatility of the alarm pheromone using enhancement of the ASR as an index of the pheromone effect. The alarm pheromone enhanced the ASR when it was presented at a distance of 10 mm but not at 200 mm. In addition, the pheromone effect was observed even after the pheromone was trapped in the adsorbent (Tenax) and then extracted using purified water. These results suggest that the alarm pheromone is both volatile and water soluble. PMID- 19135074 TI - Birth spacing in the mouse communal nest shapes adult emotional and social behavior. AB - The interactions with the mother and with peers are among the most relevant early environmental factors shaping adult brain function and behavior. In order to investigate the role of these factors, we exploited a novel early manipulation, the Communal Nest (CN), consisting in a single nest where three mothers give birth, keep their pups and share care-giving behavior from birth to weaning. In particular, we reared CD-1 swiss mice in three different CN conditions, each one characterized by a different interval between the three deliveries (Birth Spacing) of 3, 5 or 7 days (respectively, CN+/-3, CN+/-5, CN+/-7). Length of birth spacing affected maternal behavior, CN+/-7 pups receiving the highest levels. At adulthood, mice reared in the different conditions showed differences in emotional response and social skills. In the plus maze test, short birth spacing was found to be associated with enhanced emotionality, CN+/-3 mice showing highest levels of anxiety-like responses in the plus maze compared to the other two CN groups. In the social interaction test, the strategies to achieve dominance differed among the three groups. While CN+/-3 mice appeared to have a more aggressive strategy, displaying high levels of attack behavior in the first encounter, CN+/-5 and CN+/-7 mice displayed a more affiliative strategy based on social investigation. Overall, these findings show that birth spacing shapes the early mouse social environment and, in turn, affects the development of social skills and emotional responses. PMID- 19135075 TI - Signals that link energy to reproduction: gastric fill, bulk intake, or caloric intake? AB - Reproductive processes are inhibited by deficits in the availability of metabolic fuels, and this inhibition increases the chances of survival during energetic challenges and optimizes reproductive success by delaying energetically costly processes until fuels become available. The mechanisms that link energy availability to reproduction are unknown, and thus, in this study we tested the hypothesis that estrous cycles are most sensitive to sensory signals from bulk intake and gastric fill as opposed to signals from caloric intake or the availability of intracellular oxidizable fuels. The caloric content of a standard laboratory chow diet was diluted by 25, 50, or 75% with the largely indigestible fiber, cellulose, and fed to food-deprived, female hamsters throughout day 2 of the estrous cycle (ovulation and estrous behavior normally occur on day 4). The bulk intake/gastric fill hypothesis was not supported because bulk intake increased the more the diet was diluted, whereas the frequency of hamsters showing normal 4-day estrous cycles decreased with diet dilution, along with decreases in caloric intake and in plasma insulin concentrations. Rate of gastric emptying did not change significantly with diet dilution. Although consumption of a diluted diet significantly lengthened the estrous cycle, it did not affect incidence of pregnancy, litter size or pup weight. Thus, when hamsters ingest sufficient energy to support estrous behavior, they fully recover reproductive potential. In summary, neither bulk intake nor gastric fill provides critical signals necessary for reproduction, consistent with the idea that reproduction is primarily responsive fuel availability. PMID- 19135076 TI - Attenuation of cadmium-induced necrotic cell death by necrostatin-1: potential necrostatin-1 acting sites. AB - Cadmium (Cd) induces necrotic death in Chinese hamster ovary (CHO) K1 cells and we have established the responsible signaling pathway. Reportedly, necrostatin-1 (Nec-1) rescues cells from necrotic death by mediating through the death domain receptor (DR) signaling pathway. We show here that Nec-1 also effectively attenuates necrotic death triggered by Cd. Two other treatments that cause necrotic cell death, one can (z-VAD-fmk/TNF-alpha on U937 cells) and the other cannot (etherynic acid (EA) on DLD-1 cells) be rescued by Nec-1, were also studied in parallel for comparison. Results show that Nec-1 is ineffectual in modulating intracellular calcium contents, calpain activity (a downstream protease), or reactive oxygen species production. It can counteract the reduction in mitochondrial membrane potential (MMP) caused by treating CHO K1 or U937 cells with necrosis-inducing agent. However, this effect was not found in EA-treated DLD-1 cells. Notably, Nec-1 elevates NF-kappaB activity in the presence or absence of necrosis-inducing agents. Our study shows that, in addition to DR mediated necrosis, Nec-1 is effective in attenuating Cd-induced necrosis. It rescues cells with reduced MMP implying that mitochondrion is its major acting site. PMID- 19135077 TI - Face composite effects reveal abnormal face processing in Autism spectrum disorders. AB - Although it has been suggested that individuals with an Autism spectrum disorder (ASD) process faces less holistically than typically developing controls, there are few direct investigations of this hypothesis. This question was addressed before using the composite paradigm (Teunisse, J. P., & de Gelder, B. (2003). Face processing in adolescents with autistic disorder: The inversion and composite effects. Brain Cognition, 52(3), 285-294.). The results had revealed that adolescents with ASDs were less sensitive than controls to the misalignment of face parts and it was concluded their face processing was less holistic. However, because of shortcomings of the design, it was not possible to distinguish whether individuals with Autism processed both aligned and misaligned composites in a part-based fashion, or both in a holistic fashion. We compared adolescents with ASDs to controls matched on sex, age and IQ on a more complete version of the composite paradigm. The results indicate that individuals with ASDs, like controls, experience interference from facial features that they are told to ignore. However, while such interference is released for controls if parts of face composites are misaligned, individuals with ASDs show comparable interference from irrelevant parts regardless of alignment. Two different interpretations are discussed, both compatible with the idea that perceptual and or attentional abnormalities in ASDs result in a diminished level of expertise for faces. PMID- 19135078 TI - Feature mixing rather than feature replacement during perceptual filling-in. AB - 'Filling-in' occurs when a retinally stabilized object subjectively appears to vanish following perceptual fading of its boundaries. The term 'filling-in' literally means that information about the apparently vanished object is lost and replaced solely by information arising from the surrounding background. However, we find evidence that the mechanism of 'filling-in' can actually involve a process of 'feature mixing' rather than 'feature replacement,' whereby features on either side of a perceptually faded boundary merge. Here we investigate the properties of feature mixing and specify certain conditions under which such mixing occurs. Our results show that, when using visual stimuli composed of spatially alternating stripes containing different luminances or motion signals, and when using the neon-color-spreading paradigm, the filled-in luminance, motion, or color is approximately the area and magnitude weighted average of the background and the foreground luminance, motion, or color, respectively. Together, these results demonstrate that, under at least certain conditions, 'filling-in' may involve a process of feature mixing or feature averaging rather than one of feature replacement. PMID- 19135079 TI - Differential role of nitric oxide (NO) in acute and chronic stress induced neurobehavioral modulation and oxidative injury in rats. AB - The present study evaluated the effects of acute and chronic restraint stress (RS 1 h or 6 h), and their modulation by nitrergic agents on neurobehavioral and oxidative stress markers in rats. Acute RS (1 h or 6 h) reduced open arm entries (OAE) and open arm time (OAT) in the elevated plus maze test - which were attenuated by the NO precursor, L-arginine but not influenced appreciably by the NO synthase inhibitor, L-NAME. These behavioral changes were associated with differential changes in brain NO metabolites (NOx) but consistently reduced GSH and raised MDA levels in comparison to the control group. Following RS 1 h x 10 the neurobehavioral suppression and changes in brain oxidative stress markers were less pronounced as compared to the acute RS (1 h) group indicating adaptation. L-arginine pretreatment facilitated this adaptation to chronic RS (1 h). Interestingly RS 6 h x 10, induced severe behavioral suppression and aggravation of MDA and NOx levels and L-NAME pretreatment tended to protect against these chronic RS induced aggravations. These results suggest that acute and chronic RS induces duration/intensity dependent neurobehavioral and oxidative injury which are under the differential regulatory control of NO. PMID- 19135080 TI - Activation of hippocampal nitric oxide and calcium/calmodulin-dependent protein kinase II in response to Morris water maze learning in rats. AB - This study investigates the interactive roles of nitric oxide (NO) and CaM-kinase II (calcium/calmodulin-dependent protein kinase II) in Morris water maze learning. In Experiment I, experimental rats received 5 days of training on a Morris water maze, where the controls were trained in the water maze with no spatial cue condition or were trained via a visually guided landmark condition. The experimental rats showed improvement in their rate of spatial learning in the water maze. The escape latencies were significantly correlated with the Ca2+ independent activity of the hippocampal CaM-kinase II. Moreover, there was a significant increase in the endogenous phosphorylation of neuronal NOS and CaM kinase II in the experimental group when compared to the controls. The intra hippocampal infusion of 7-NI, KN-93, or AP5 did disrupt water maze learning. SDS PAGE analysis showed that these drugs significantly depressed phosphorylation of hippocampal NOS. The Ca2+-independent activity of hippocampal CaM-kinase II was significantly lower in the KN-93 or the AP5 infused group when compared to the controls. Although these depressed activities were not reversed by the infusion of NO donor (sodium nitroprusside, SNP), the rats' water maze learning behavior were ameliorated significantly. These results, taken together, indicate that the NOS activation is essential for water maze learning, which may be triggered via the CaM-kinase II activation in hippocampus. PMID- 19135081 TI - Medical students' self-perceived competence and prescription of patient-centered physical activity. AB - OBJECTIVE: The primary objective of this study was to explore medical students' perceptions of their own competence and the importance they assign to patient centered physical activity (PA) prescription. METHODS: 246 undergraduate medical students (27% response rate) from two large universities in Western Canada completed an online survey designed to assess their perceived competence and importance related to patient-centered PA prescription. Data collection took place in September and October of 2007. RESULTS: While medical students perceived PA-related prescription to be important (M(response)=26.6 out of 36, SD=5.1), students perceived they had only moderate competence at conducting PA-related prescription (M(response)=20.7 out of 36, SD=6.8). Students achieving national PA guidelines perceived significantly higher competence than students not achieving PA guidelines. Students in their first or second year of medical school perceived PA-related prescription to be of higher importance than students in their third or fourth years. CONCLUSION: Medical students indicated that patient-centered PA prescription was important. However, they indicated less than moderate competence at performing several fundamental PA prescription behaviors. This study suggests that medical students may not be adequately prepared to dispense patient-centered PA prescriptions with their patients. PMID- 19135082 TI - Hydrolytic fragmentation of seed gums under microwave irradiation. AB - The seed gum solutions of Ipomoea purga, Ipomoea palmata, Ipomoea dasysperma, Cyanaposis tetragonolobus (Guar gum) and Crotolaria medicaginea were microwave (MW) irradiated and their degradation to oligo and monosaccharides was investigated. The gum solutions were fragmented into oligosaccharides/constituent monosaccharides depending upon the length of MW exposure in presence of catalytic amount of mineral acid or even when no acid was used. A mechanism for the microwave induced hydrolytic degradation of the seed gums has been proposed. The MW exposure time required for the partial and complete degradation of the gums was found dependent on the types of the linkages and degree of the branching present in the gums. PMID- 19135084 TI - Quantitative microdialysis using modified ultraslow microdialysis: direct rapid and reliable determination of free brain concentrations with the MetaQuant technique. AB - The only method to quantify free extracellular levels of drugs in the brain of living animals is microdialysis. However, quantitative microdialysis has been hampered by methodological issues for decades. The problems arise from the need to establish the in vivo recovery for appropriate quantitation. In dealing with these issues the "dynamic no-net-flux" (DNNF) method seemed to be the experimental method of choice. Major disadvantages were, however, the need for a very high degree of bioanalytical precision and accuracy and the need for a large number of animals. Moreover, today we know that the experimental data are not always straightforward. To improve robustness and practicality of quantitative microdialysis sampling we modified the ultraslow microdialysis approach. Ultraslow microdialysis uses very low microdialysis flow rates (<200 nl/min) which increase recovery (both in vivo and in vitro) to over 90%. However, new practical issues arise when attempting to work with these flow rates. The resulting very low volumes and long lag times make this method very impractical for general application. In the modified version, addition of a carrier flow after the dialysis process has been completed, which negates the problems of long lag times and low volumes. The resulting dilution of the dialysis sample concentration can simply be mathematically corrected. In the current study we measured the free brain levels of two CNS compounds using the classic DNNF and the new modified ultraslow dialysis method. Modified ultraslow microdialysis was shown to generate robust data with the use of only small numbers of rats. The method is a promising tool for common straightforward screening of blood-brain barrier penetration of compounds into the brain. PMID- 19135083 TI - Method for the construction and use of carbon fiber multibarrel electrodes for deep brain recordings in the alert animal. AB - Microiontophoresis of neuroactive substances during single unit recording in awake behaving animals can significantly advance our understanding of neural circuit function. Here, we present a detailed description of a method for constructing carbon fiber multibarrel electrodes suitable for delivering drugs while simultaneously recording single unit activity from deep structures, including brainstem nuclei and the cerebellum, in the awake behaving primate. We provide data that should aid in minimizing barrel resistance and the time required to fill long, thin multibarrel electrodes with solutions. We also show successful single unit recording from a variety of areas in the awake squirrel monkey central nervous system, including the vestibular nuclei, Interstitial Nucleus of Cajal, and the cerebellum. Our descriptions and data should be useful for investigators wishing to perform single unit recordings during microiontophoresis of neuroactive substances, particularly in deep structures of animals with chronically implanted recording chambers. PMID- 19135085 TI - Simultaneous telemetric monitoring of tail-skin and core body temperature in a rat model of thermoregulatory dysfunction. AB - Temperature dysfunction, clinically described as hot flashes/flushes and night sweats, commonly occur in women transitioning through menopause. Research in this field has yet to fully elucidate the biological underpinnings explaining this dysfunction. The need to develop animal models that can be used to study hormone dependent temperature regulation is essential to advancing this scientific area. Development of telemetric transmitters for monitoring tail-skin (TST) and core body (CBT) temperatures for animal research has increased the accuracy of data by reducing extraneous factors associated with previous methods. However, until recently, TST and CBT could not be simultaneously measured telemetrically within the same animal. In this report, new dual temperature monitoring transmitters were validated by simultaneously evaluating them with the single measurement transmitters using the ovariectomized (OVX) rat thermoregulatory dysfunction model. A major advantage of measuring TST and CBT in the same animal is the ability to relate temporal changes on these two temperature parameters. Comparative experimentation was performed by single administration of clonidine (alpha(2) adrenergic agonist), MDL-100907 (5-HT(2a) antagonist), or a 7-day treatment of 17alpha-ethinyl estradiol (EE). Clonidine caused decreases in TST and CBT, MDL-100907 caused increases in TST while decreasing CBT, and EE caused decreases in TST with minor CBT decreases only at the higher dose. Data from either probe type showed similar results on temperature parameters regardless of transmitter used. These findings support the use of the new dual temperature transmitters and should enhance the quality and interpretation of data being generated in thermoregulation studies. PMID- 19135086 TI - Biocybernetics of attention in the tinnitus decompensation: An integrative multiscale modeling approach. AB - Tinnitus is one of the most common symptoms affecting people all over the world. In the absence of an established cure many individuals are not only faced with the need to adjust to the sensation of the tinnitus noise, but also with psychological comorbidities. In recent years, different studies have been directed to elucidate the psychophysiological mechanisms that are involved in the tinnitus decompensation. From these, special emphasis has been placed on studies related to attention and habituation, which accordingly play a crucial role in current tinnitus therapy approaches. In spite of such progress, the relationship between selective attention and the tinnitus decompensation with respect to large scale neural correlates is still not well understood. In order to address this issue, we propose an integrative multiscale modeling approach for studying neural correlates of auditory selective attention in the tinnitus decompensation. Computational simulations based on our model confirmed electroencephalographic human data of both auditory selective attention and the tinnitus decompensation. It is concluded that the proposed methodology represents a promising approach to give insight into the neurodynamics of auditory selective attention in the tinnitus decompensation. PMID- 19135087 TI - High-resolution fMRI: overcoming the signal-to-noise problem. AB - Increasing the spatial resolution in functional Magnetic Resonance Imaging (fMRI) inherently lowers the signal-to-noise ratio (SNR). In order to still detect functionally significant activations in high-resolution images, spatial smoothing of the data is required. However, conventional non-adaptive smoothing comes with a reduced effective resolution, foiling the benefit of the higher acquisition resolution. We show how our recently proposed structural adaptive smoothing procedure for functional MRI data can improve signal detection of high-resolution fMRI experiments regardless of the lower SNR. The procedure is evaluated on human visual and sensory-motor mapping experiments. In these applications, the higher resolution could be fully utilized and high-resolution experiments were outperforming normal resolution experiments by means of both statistical significance and information content. PMID- 19135088 TI - Analyzing the interaction of pathogens with the host immune system. AB - Infectious diseases continue to represent a great burden to human society, being responsible for approximately 14 to 16 million deaths annually. Today, Mycobacterium tuberculosis, the causative agent of tuberculosis, remains one of the most important infectious agents. Moreover, with the emergence of multi drug and extensive drug resistant strains of M. tuberculosis, there is a great need for a better understanding of the virulence strategies utilized by this pathogen. In this manuscript, we discuss some of the strategies that have been followed to unravel virulence mechanisms utilized by M. tuberculosis. Knowledge of these mechanisms is important to reveal potential targets that may be useful for the development of compounds to treat tuberculosis. PMID- 19135089 TI - Performance characteristics of a quantitative, standardised varicella zoster IgG time resolved fluorescence immunoassay (VZV TRFIA) for measuring antibody following natural infection. AB - Infection by Varicella Zoster virus (VZV) during pregnancy has been associated with adverse foetal development and more severe disease in the mother. Accurate determination of VZV immunity in pregnant women exposed to VZV, with no history of chickenpox, guides therapeutic interventions. The accepted gold standard assay for the determination of immunity/protection against Varicella Zoster virus was for many years the fluorescent antibody to membrane antigen (FAMA) assay which is labour intensive and subjective. A validated alternative is the Merck glycoprotein EIA (Merck Sharp & Dohme Research Laboratories, West Point, PA, USA) which reports VZV IgG levels in enzyme units per ml (EU/ml) because an internal, non-international reference serum is used as calibrator. Comparison of different VZV IgG detection assays is hampered by a lack of an agreed cut-off in standardised units. A time resolved fluorescence immunoassay (TRFIA) for VZV IgG using British Standard VZV antibody has been developed and standardised. The limit of detection of VZV IgG by this assay was of the order 39-78mIU/ml. Following comparison with the Merck glycoprotein EIA and the application of the USA Advisory Committee on Immunization Practices recommended 5.0EU/ml cut-off the following standardised cut-offs in mIU/ml are proposed. A VZV TRFIA IgG cut-off of less than 100mIU/ml VZV IgG equates with susceptibility and an equivocal range of 100mIU/ml to less than 150mIU/ml is proposed. VZV IgG levels of 150mIU/ml, or greater, are indicative of natural infection at some time and the ability to mount a protective immune response is inferred. PMID- 19135091 TI - Effective siRNA targeting of the 3' untranslated region of the West Nile virus genome. AB - West Nile virus (WNV) is an emerging human pathogen for which specific antiviral therapy has not been developed. The therapeutic potential of RNA interference (RNAi) as a sequence-specific inhibitor of WNV has been well demonstrated. Although multiple siRNA targets have been identified within the genomic coding region, targets within the untranslated regions (UTR), which encode cis-acting regulatory elements, remain relatively unknown. In WNV and other flaviviruses, UTRs are located at the genomic termini. These regions form complex secondary structures, which pose difficulty when designing effective siRNA targets. In this study, we report the identification of siRNA targets in the WNV 3' UTR. These targets were selected by siRNA predictor algorithms, and synthesized as short hairpin RNA sequences from a plasmid-based expression system. Vero cells stably expressing these sequences had greatly diminished ability to support WNV replication but not the related dengue virus, demonstrating that the siRNAs were effective and suppressed WNV viral replication in a sequence-specific manner. The siRNAs developed in this study could function as potential antiviral therapeutics and as genetic tools to investigate the role of 3' UTR in WNV pathogenesis. PMID- 19135090 TI - Using high-throughput genomics to study hepatitis C: what determines the outcome of infection? AB - High-throughput genomic methods are now being used to study a wide variety of viral diseases, in an effort to understand how host responses to infection can lead either to efficient elimination of the pathogen or the development of severe disease. This article reviews how gene expression studies are addressing important clinical issues related to hepatitis C virus infection, in which some 15-25% of infected individuals are able to clear the virus without treatment, while the remainder progress to chronic liver disease that can lead to cirrhosis and death. Similar methods are also being used in an effort to identify the mechanisms underlying the failure of some hepatitis C patients to respond to interferon-alpha/ribavirin therapy. By providing a detailed picture of virus-host interactions, high-throughput genomics could potentially lead to the identification of novel cellular targets for the treatment of hepatitis C. PMID- 19135092 TI - Validation of a novel social investigation task that may dissociate social motivation from exploratory activity. AB - The present series of studies provide validation of a new paradigm that uniquely combines the assessment of the propensity to engage in social investigation with measures of (nonsocial) exploratory activity in rats. Assessment of this social investigation paradigm indicated that (a) rats showed a robust preference for social investigation over nonsocial exploratory activity, (b) female rats showed a greater preference for social investigation than male rats, (c) no signs of habituation in these responses were observed when rats were tested once daily for 4 consecutive days, (d) the preference for social investigation was stable and robust in both the dark and light periods of the daily light cycle for 5 consecutive days, and (e) testing under bright light conditions suppressed social investigation. In addition, acute administration of opiate drugs, low dose morphine (1.0 mg/kg) and naltrexone (1.0 mg/kg) produced a more robust attenuation of social investigation than nonsocial exploratory activity. Amphetamine increased both forms of investigation and haloperidol had the opposite effect, but the overall preference for social investigation over exploratory activity remained largely intact after both amphetamine and haloperidol injection. Together, these findings validate the use of this behavioral task to assess changes in social-motivation and general exploratory activity. Importantly, the task is bi-directionally sensitive to subject characteristics (i.e., sex), drug manipulations which modulate social motivation, and environmental manipulations. PMID- 19135094 TI - Entamoeba histolyticaEhGEF1 structure and mutational analysis: New specific residues critical for function. AB - This paper reports the EhGEF1-EhRacG and EhGEF1-EhRho1 molecular complexes from Entamoeba histolytica. The not conserved amino acids Gln201,Tyr299, Gln302, Lys312, Asn313, Phe314 and Ile324 were localized, by means of an in silico computational analysis, at the interface of the exposed face from the DH domain of EhGEF1, which are important to establish the contact with its target GTPases EhRacG and EhRho1. Functional studies of nucleotide exchange of Phe314Ala mutant showed a decrement of 80% on EhRacG GTPase; in contrast the Ile324Ala mutant exhibited a reduction of 77%, specifically on EhRho1; meanwhile the Gln302Ala mutant showed a reduction of approximately 50% on the exchange activity for both GTPases. Moreover, the functional studies of the protein EhGEF1 mutants in the conserved residues Thr194Ala, Asn366Ala and Glu367Ala indicated that contrary to what has been reported for other systems, the mutation of these residues did not alter considerably its catalytic activity. PMID- 19135095 TI - Specific modulation of protein kinase activity via small peptides. AB - Protein kinases are key drug targets for several diseases. However, despite large efforts, the development of drugs that are specific for these kinases has been a great challenge. The discovery that protein-protein interactions can be mediated by small linear sequences has invigorated the bioinformatics field and there are increasing efforts to create computer programs that identify and characterize linear sequences responsible for protein-protein interactions. The development of peptides as modulators of target protein activity is facilitated by in silico approaches. There are numerous applications for peptide modulators of protein protein interactions and peptide modulators of kinase activity. Peptide modulators may be used either as tools to elucidate specific signaling pathways, as drug leads, or as drugs themselves. In the present review we will discuss the development of such peptide modulators and some of their applications. PMID- 19135093 TI - Stimulus-response and response-outcome learning mechanisms in the striatum. AB - While midbrain DA neurons show phasic activations in response to both reward predicting and salient non-reward events, activation responses to primary and conditioned rewards are sustained for several hundreds of milliseconds beyond those elicited by salient non-reward-related stimuli. The longer-duration DA reward response and corresponding elevated DA release in striatal target sites may selectively strengthen currently-active corticostriatal synapses, i.e., those associated with the successful reward-procuring behavior. This paper describes how similar models of DA-mediated plasticity of corticostriatal synapses may describe both stimulus-response and response-outcome learning. DA-mediated strengthening of corticostriatal synapses in regions of the dorsolateral striatum receiving afferents from primary sensorimotor cortex is likely to bind corticostriatal inputs representing the previously-emitted movement to striatal outputs contributing to the selection of the next movement segment in a behavioral sequence. Within the striatum, more generally, inputs from distinct regions of the frontal cortex that code independently for movement direction and reward expectation send convergent projections to striatal output cells. DA mediated strengthening of active corticostriatal synapses promotes the future output of the striatal cell under similar input conditions. This is postulated to promote persistence of neuronal activity in the very cortical cells that drive corticostriatal input, leading to the establishment of sustained reverberatory loops that permit cortical movement-related cells to maintain activity until the appropriate time of movement initiation. PMID- 19135096 TI - FGF-activated calcium channels control neural gene expression in Xenopus. AB - In vertebrates, the formation of the nervous system starts at gastrulation with a process called neural induction. This process requires, at least in part, the inhibition of BMP signalling in the ectoderm by noggin, as well as FGF receptor activation and Ca2+ signalling. Our studies with Xenopus embryos suggest that an increase in intracellular Ca2+ concentration ([Ca2+]i), via dihydropyridine sensitive Ca2+ channels (DHP-sensitive Ca2+ channels) is necessary and sufficient to direct the ectodermal cells toward a neural fate, and that Ca2+ directly controls the expression of neural genes. The mechanism by which the DHP-sensitive Ca2+ channels are activated during neural induction remains unknown. One possible mechanism is via the activation of FGF signalling. Using isolated ectoderm tissue, here we demonstrated that FGF-4 depolarises the membrane of ectodermal cells and induces an increase in [Ca2+]i. This Ca2+ increase can be blocked by SU5402, an FGF receptor inhibitor, and by DHP-sensitive Ca2+ channel antagonists. These inhibitors also block the induction of neural genes. We discuss a possible gating mechanism for the activation of DHP-sensitive Ca2+ channels via the FGF signalling pathway, which involves arachidonic acid and TRPC1 channel activation. PMID- 19135097 TI - Development of a quantitative real-time PCR assay for detection of Vibrio tubiashii targeting the metalloprotease gene. AB - Vibrio tubiashii has recently re-emerged as a pathogen of bivalve larvae, causing a marked increase in the mortality of these species within shellfish rearing facilities. This has resulted in substantial losses of seed production and thus created the need for specific as well as sensitive detection methods for this pathogen. In this project, quantitative PCR (qPCR) primers were developed and optimized based upon analysis of the V. tubiashii vtpA gene sequence, encoding a metalloprotease known to cause larval mortality. Standard curves were developed utilizing dilutions of known quantities of V. tubiashii cells that were compared to colony forming unit (CFU) plate counts. The assay was optimized for detection of vtpA with both lab-grown V. tubiashii samples and filter-captured environmental seawater samples seeded with V. tubiashii. In addition, the primers were confirmed to specifically detect only V. tubiashii when tested against a variety of non-target Vibrio species. Validation of the assay was completed by analyzing samples obtained from a shellfish hatchery. The development of this rapid and sensitive assay for quantitative detection of V. tubiashii will accurately determine levels of this bacterium in a variety of seawater samples, providing a useful tool for oyster hatcheries and a method to assess the presence of this bacterium in the current turbulent ocean environment. PMID- 19135098 TI - Domestic wastewater influent profiling using mitochondrial real-time PCR for source tracking animal contamination. AB - Real-time PCR amplifying mammalian and avian mitochondrial DNA (mtDNA) was developed to characterize domestic and light industrial wastewater influent from two municipal wastewater treatment facilities (WWTF) over a 24-week period. Influent samples were assayed with species-specific primers and dual-labeled probes for human, bovine, swine, dog, cat, Canada goose and white-tailed deer to detect and quantify eukaryotic mtDNA contributors to local municipal wastewaters. Human (mean=9.6 x 10(4) copies/ml) and dog (mean=5.3 x 10(2) copies/ml) mtDNA were detected in all 24 samples (12 samples/site). Bovine and swine mtDNA were detected sporadically and at lower levels than human mtDNA, means=3.0 x 10(4) and 9.5 x 10(2) copies/ml, respectively. Domestic cat, Canada goose and white-tailed deer were detected only once in 24 samples. Mitochondrial DNA concentrations were compared to other bacterial, chemical and spectrophotometric parameters. Human mtDNA was positively correlated with ammonia concentration (P=0.01) and initial OD(600) reading (P=0.02) at one WWTF. Bovine mtDNA was positively correlated with biological oxygen demand (BOD) (P=0.02), final DNA concentration (P=0.03), initial and final humic acid concentrations (P=0.01, P=0.01), and final OD(600) (P=0.03) at one WWTF and total suspended solids (TSS) (P=0.04, P=0.09) at both facilities. Fecal coliforms were not positively or negatively correlated with mtDNA concentrations of any species assayed. For source tracking purposes, a combination of human (10(5) copies/ml) and dog mtDNA signal (10(2) copies/ml) could be indicative of municipal domestic wastewater contamination of environmental waters. PMID- 19135099 TI - Assessment of horizontal gene transfer in Lactic acid bacteria--a comparison of mating techniques with a view to optimising conjugation conditions. AB - Plate, filter and broth mating techniques were assessed over a range of pHs using three Lactococcus lactis donor strains (one with an erythromycin resistance marker and two with tetracycline resistance markers, all located on transferable genetic elements) and one L. lactis recipient strain. Transconjugants were confirmed using antibiotic selection, E-tests to determine MICs, PCR assays to detect the corresponding marker genes, DNA fingerprinting by pulsed-field gel electrophoresis (PFGE), and Southern blotting. Horizontal gene transfer (HGT) rates varied (ranging from 1.6 x 10(-1) to 2.3 x 10(-8)). The general trend observed was plate > filter > broth, independent of pH. Our data suggests that standardisation of methodologies to be used to assess HGT, is warranted and would provide a meaningful assessment of the ability of commensal and other bacteria in different environments to transfer relevant markers. PMID- 19135100 TI - Mental fatigue and impaired response processes: event-related brain potentials in a Go/NoGo task. AB - The effects of mental fatigue on the availability of cognitive resources and associated response-related processes were examined using event-related brain potentials. Subjects performed a Go/NoGo task for 60 min. Reaction time, number of errors, and mental fatigue scores all significantly increased with time spent on the task. The NoGo-P3 amplitude significantly decreased with time on task, but the Go-P3 amplitude was not modulated. The amplitude of error-related negativity (Ne/ERN) also decreased with time on task. These results indicate that mental fatigue attenuates resource allocation and error monitoring for NoGo stimuli. The Go- and NoGo-P3 latencies both increased with time on task, indicative of a delay in stimulus evaluation time due to mental fatigue. NoGo-N2 latency increased with time on task, but NoGo-N2 amplitude was not modulated. The amplitude of response locked lateralized readiness potential (LRP) significantly decreased with time on task. Mental fatigue appears to slows down the time course of response inhibition, and impairs the intensity of response execution. PMID- 19135102 TI - Generation of tetracycline-inducible conditional gene knockout cells in a human Nalm-6 cell line. AB - Conditional gene knockout by homologous recombination combined with an inducible gene expression system is a powerful approach for studying gene function, although homologous recombination in human cells occurs infrequently. The tetracycline-regulated gene expression (Tet-Off) system is a convenient method for achieving conditional gene knockout, but it is not always promising in Nalm 6, a rare human cell line highly effective for gene targeting. Here we modified the Tet-Off system and applied it to the Nalm-6 cell line successfully by using an internal ribosome entry site to drive a selectable marker from the same tetracycline-responsive promoter for the transgene. We also inserted the gene for the tetracycline-controlled transactivator under the control of a potent CAG promoter. These modifications enabled us to easily obtain rare clones that express optimal amounts of tetracycline-regulated transgenes. We thereby generated a 'tetracycline-inducible conditional gene knockout' for the proliferation-associated SNF2-like gene (PASG) in a Nalm-6 cell line, in which the expression of PASG can be depleted in a tetracycline-dependent manner on a knockout background. This method is applicable to any human genes, making this gene-targeting system using the Nalm-6 cell line a promising tool for analyzing gene function. PMID- 19135101 TI - Expression of mitochondrial complex I subunit gene NDUFV2 in the lymphoblastoid cells derived from patients with bipolar disorder and schizophrenia. AB - Several studies have suggested mitochondrial abnormality in bipolar disorder (BD) and schizophrenia (SZ). We have previously reported the decreased expression of mitochondrial complex I subunit gene, NDUFV2 at 18p11, in lymphoblastoid cell lines (LCLs) from Japanese patients with bipolar I disorder (BDI). Recently it was reported that no differences were found in NDUFV2 mRNA levels in LCLs of Caucasian BDI patients compared with controls. In this study, we tested the altered expression of NDUFV2 in extended Japanese LCLs and LCLs from different ethnic groups. Similar tendency was found in the current study compared with our previous study, since decreased expression of NDUFV2 in LCLs from Japanese patients with BDI was found (p=0.03). We also found that the expressions of NDUFV2 were up-regulated in those from patients with Japanese bipolar II disorder (p=0.001) and the mRNA levels of this gene were down-regulated in Caucasian SZ (p=0.000001) compared with controls. Furthermore, we revealed that the mRNA expression of NDUFV2 in LCLs cultured with valproate, one of mood stabilizers, were significantly increased compared with controls (p=0.02). Our study presented the further evidence of biological significance of NDUFV2 in BD and SZ. PMID- 19135103 TI - High-level expression of rabies virus glycoprotein with the RNA-based Semliki Forest Virus expression vector. AB - Rabies is to this date one of the most important death causing zoonotic viral diseases, with 98% of deaths reported in developing countries, where access to modern vaccines and tools for efficient diagnostic remain unaffordable. In this paper, we describe a newly engineered RNA-based rabies virus glycoprotein (RVGP) expression vector based on the Semliki Forest Virus (SFV) system. A recombinant SFV carrying an RNA coding for RVGP (SFV-RVGP) was constructed and the RVGP expression was evaluated in animal cell cultures. The mRNA coding for RVGP and the RVGP itself were assessed by qPCR, Western-blotting, confocal microscopy, flow cytometry and ELISA. Moreover, SFV-RVGP was proven to be highly efficient in expressing the functionally trimeric RVGP. SFV-RVGP is easy to produce and efficient in different cell lines, making it an interesting candidate for efficient and functional viral glycoprotein expression. PMID- 19135104 TI - A peptide-targeted delivery system with pH-sensitive amphiphilic cell membrane disruption for efficient receptor-mediated siRNA delivery. AB - The efficient delivery of therapeutic siRNA into cells of interest is a critical challenge to broad application of RNAi. In this study, we developed a peptide targeted delivery system for highly efficient receptor-mediated cellular siRNA delivery. The targeted delivery system was readily prepared by in situ functionalization of a polymerizable pH-sensitive amphiphilic surfactant, N-(1 aminoethyl)iminobis[N-(oleicyl-cysteinyl-histinyl-1-aminoethyl)propionamide] (EHCO) and self-assembly with siRNA. The intrinsic pH-sensitive amphiphilicity of EHCO at pH 5-6 was able to induce cell membrane disruption at endosomal pH and facilitate endosomal escape of the siRNA nanoparticles after internalization. The siRNA/EHCO nanoparticles and PEGylated siRNA/EHCO nanoparticles were not cytotoxic as compared to PEI/siRNA or TransFast/siRNA nanoparticles. siRNA/EHCO nanoparticles resulted in higher siRNA delivery efficiency than PEI and TransFast. The PEGylation of the siRNA/EHCO nanoparticles significantly reduced non-specific cell uptake. The incorporation of a bombesin peptide via a PEG spacer resulted in specific cellular uptake and high gene silencing efficiency in CHO-d1EGFP cells with overexpression of bombesin receptors. Receptor-mediated endocytosis and pH-sensitive amphiphilic endosomal escape are the advantageous features of the targeted siRNA delivery system for highly efficient cell-specific siRNA delivery. This novel targeted delivery system holds a great promise for systemic and targeted delivery of therapeutic siRNA. PMID- 19135105 TI - Ins and outs of the ABCG2 multidrug transporter: an update on in vitro functional assays. AB - The major aim of this chapter is to provide a critical overview of the in vitro methods available for studying the function of the ABCG2 multidrug transporter protein. When describing the most applicable assay systems, in each case we present a short overview relevant to ABC multidrug transporters in general, and then we concentrate on the tools applicable to analysis of substrate-drug interactions, the effects of potential activators and inhibitors, and the role of polymorphisms of the ABCG2 transporter. Throughout this chapter we focus on recently developed assay systems, which may provide new possibilities for analyzing the pharmacological aspects of this medically important protein. PMID- 19135106 TI - QSAR analysis and molecular modeling of ABCG2-specific inhibitors. AB - In addition to its critical role is controlling drug availability and protecting sensitive organs and stem cells through cellular detoxification, breast cancer resistance protein (BCRP/ABCG2) plays an important role in cancer cell resistance to chemotherapy, together with P-glycoprotein/ABCB1. A main approach to abolish multidrug resistance is to find out specific inhibitors of the drug-efflux activity, able to chemosensitize cancer cell proliferation. Many efforts have been primarily focused on ABCB1, discovered thirty years ago, whereas very few studies have concerned ABCG2, identified much more recently. This review describes the main types of inhibitors presently known for ABCG2, and how quantitative structure-activity relationship analysis among series of compounds may lead to build up molecular models and pharmacophores allowing to design lead inhibitors as future candidates for clinical trials. A special attention is drawn on flavonoids which constitute a structurally-diverse class of compounds, well suited to identify potent ABCG2-specific inhibitors. PMID- 19135107 TI - Barrier properties of mucus. AB - Mucus is tenacious. It sticks to most particles, preventing their penetration to the epithelial surface. Multiple low-affinity hydrophobic interactions play a major role in these adhesive interactions. Mucus gel is also shear-thinning, making it an excellent lubricant that ensures an unstirred layer of mucus remains adherent to the epithelial surface. Thus nanoparticles (NP) must diffuse readily through the unstirred adherent layer if they are to contact epithelial cells efficiently. This article reviews some of the physiological and biochemical properties that form the mucus barrier. Capsid viruses can diffuse through mucus as rapidly as through water and thereby penetrate to the epithelium even though they have to diffuse 'upstream' through mucus that is being continuously secreted. These viruses are smaller than the mucus mesh spacing, and have surfaces that do not stick to mucus. They form a useful model for developing NP for mucosal drug delivery. PMID- 19135108 TI - Pharmacokinetics and pharmacogenomics in esophageal cancer chemoradiotherapy. AB - Esophageal cancer is one of the most lethal malignancies. Surgical resection of the tumor from the primary site has been the standard treatment, especially for localized squamous cell carcinoma, but considerable clinical efforts during the last decade have resulted in novel courses of treatment. These options include chemoradiotherapy, consisting of a continuous infusion of 5-fluorouracil (5-FU), cisplatin (CDDP), and concurrent radiation. Given the substantial inter- and/or intra-individual variation in clinical outcome, future improvements will likely require the incorporation of a novel anticancer drug, pharmacokinetically guided administration of CDDP or 5-FU, and identification of potential responders by patient genetic profiling prior to treatment. In this review, the latest information on incidence, risk factors, biomarkers, therapeutic strategies, and the pharmacokinetically guided or genotype-guided administration of CDDP and 5-FU is summarized for future individualization of esophageal cancer treatment. PMID- 19135110 TI - Willingness to use functional breads. Applying the Health Belief Model across four European countries. AB - The present study focused on the role of the Health Belief Model (HBM) in predicting willingness to use functional breads, across four European countries: UK (N=552), Italy (N=504), Germany (N=525) and Finland (N=513). The behavioural evaluation components of the HBM (the perceived benefits and barriers conceptualized respectively as perceived healthiness and pleasantness) and the health motivation component were good predictors of willingness to use functional breads whereas threat perception components (perceived susceptibility and perceived anticipated severity) failed as predictors. This result was common in all four countries and across products. The role of 'cue to action' was marginal. On the whole the HBM fit was similar across the countries and products in terms of significant predictors (the perceived benefits, barriers and health motivation) with the exception of self-efficacy which was significant only in Finland. Young consumers seemed more interested in the functional bread with a health claim promoting health rather than in reducing risk of disease, whereas the opposite was true for older people. However, functional staple foods, such as bread in this European study, are still perceived as common foods rather than as a means of avoiding diseases. Consumers seek these foods for their healthiness (the perceived benefits) as they expect them to be healthier than regular foods and for the pleasantness (the perceived barriers) as they do not expect any change in the sensory characteristics due to the addition of the functional ingredients. The importance of health motivation in willingness to use products with health claims implies that there is an opening for developing better models for explaining health-promoting food choices that take into account both food and health-related factors without making a reference to disease-related outcome. PMID- 19135111 TI - "Smart Bodies" school wellness program increased children's knowledge of healthy nutrition practices and self-efficacy to consume fruit and vegetables. AB - Diets rich in fruit and vegetables are important for long-term health yet children frequently do not like these foods. The "Smart Bodies" school wellness program sought to increase children's knowledge of healthy nutritional practices, improve psychosocial variables associated with eating fruit and vegetables, and develop preferences for these foods. A randomized controlled intervention trial was conducted in 14 low-income, urban, public elementary schools (seven pairs). Data from 278 fourth and 282 fifth graders (234 boys, 326 girls; 82% Black, 10% White, 1% Hispanic, 5% Asian, 2% Other) were examined using multi-level modeling. The 12-week intervention program included participation in an interactive wellness exhibit and a classroom curriculum that emphasized consumption of fruit and vegetables. After the intervention, children that participated in the "Smart Bodies" program had greater nutrition knowledge and expressed more confidence that they could eat fruit instead of a favorite dessert, drink fruit juice and consume the recommended number of fruits and vegetables servings each day. Preferences for fruit and vegetables did not change as a result of participating in the program. These findings demonstrate that the "Smart Bodies" school-based wellness intervention positively impacted children's nutrition knowledge and psychosocial variables associated with consuming fruit and vegetables. PMID- 19135112 TI - The relationship between stress, dietary restraint, and food preferences in women. AB - The relationship between stress, restraint, and eating has been studied using various methods, including retrospective self-reports of stress and eating that are open to inaccuracies. Additionally, laboratory research has not systematically varied the fat content and sweetness of food items to assess how stress relates to specific food preferences. In this study of 40 women we examined the role of restraint and experimentally induced stress on the amount of sweet, salty, high-fat, and low-fat food consumed. High-restraint women ate more high-fat food than did low-restraint women, regardless of stress level. High stressed women preferred sweet, high-fat food more than did low-stressed women, whereas low-stressed women ate more low-fat than high-fat food. There was no interaction between restraint and stress level. Social influence effects of small group testing may have increased the ego-threat of the stressor or disinhibited high-restraint women in both stress groups. Future laboratory research is needed to assess the role of the presence of others in both stress induction and eating behaviors. PMID- 19135109 TI - ABCG2: a perspective. AB - ABCG2, or breast cancer resistance protein (BCRP), is an ABC transporter that has been the subject of intense study since its discovery a decade ago. With high normal tissue expression in the brain endothelium, gastrointestinal tract, and placenta, ABCG2 is believed to be important in the protection from xenobiotics, regulating oral bioavailability, forming part of the blood-brain barrier, the blood-testis barrier, and the maternal-fetal barrier. Notably, ABCG2 is often expressed in stem cell populations, where it likely plays a role in xenobiotic protection. However, clues to its epigenetic regulation in various cell populations are only beginning to emerge. While ABCG2 overexpression has been demonstrated in cancer cells after in vitro drug treatment, endogenous ABCG2 expression in certain cancers is likely a reflection of the differentiated phenotype of the cell of origin and likely contributes to intrinsic drug resistance. Notably, research into the transporter's role in cancer drug resistance and its development as a therapeutic target in cancer has lagged. Substrates and inhibitors of the transporter have been described, among them chemotherapy drugs, tyrosine kinase inhibitors, antivirals, HMG-CoA reductase inhibitors, carcinogens, and flavonoids. This broad range of substrates complements the efficiency of ABCG2 as a transporter in laboratory studies and suggests that, while there are redundant mechanisms of xenobiotic protection, the protein is important in normal physiology. Indeed, emerging studies in pharmacology and toxicology assessing polymorphic variants in man, in combination with murine knockout models have confirmed its dynamic role. Work in pharmacology may eventually lead us to a greater understanding of the physiologic role of ABCG2. PMID- 19135113 TI - Heterogeneous expression of HLA-G1, -G2, -G5, -G6, and -G7 in myeloid and plasmacytoid dendritic cells isolated from umbilical cord blood. AB - Human leukocyte antigen (HLA)-G is a human nonclassic major histocompatibility complex (MHC) molecule characterized by a limited polymorphism and a low, restricted cell surface expression. HLA-G is constitutively expressed on trophoblasts, fetal endothelial, and epithelial cells, conferring alloimmune protection during pregnancy. HLA-G is also expressed in some malignancies and on macrophages and dendritic cells (DC) in tumoral and inflammatory diseases. Because DC constitute an important component in the immune response and umbilical cord blood has a different immune behavior than peripheral blood, the HLA-G protein profile and mRNA expression were investigated on the different DC subsets present in cord blood. Surface and intracellular expression have been reported on DC and HLA-G1, -G2, -G5, -G6, and -G7 transcripts were present. Different levels of soluble HLA-G were obtained from serum and correlated with gene expression. These data are in contrast with the data previously described for adult peripheral blood, where a limited pattern of HLA-G transcripts was reported; only in the maturation process were more isoforms present. These results demonstrate that DC from cord blood have a different behavior than DC in peripheral blood and could be in accordance with the results obtained in cord blood transplantation, where a lesser effect of graft-versus-host disease exists than in bone marrow transplantation. PMID- 19135114 TI - Toll-like receptor 4 gene polymorphism and early onset of diabetic retinopathy in patients with type 2 diabetes. AB - Toll-like receptor 4 (TLR4) is an important mediator of innate immunity. Type 2 diabetes (DM2) might be associated with changed innate immune response. We investigated whether the polymorphisms in the TLR4 gene are associated with diabetic retinopathy (DR). The study group of 864 patients with DM2 and 420 healthy individuals were genotyped. In the patient group 352 subjects were diagnosed with DR. Out of the remaining 512, 140 had DM2 for > or = 10 years but no DR. In the DM2 group 7.4% of patients were heterozygous for the Asp299Gly polymorphism compared with 6.5% controls. For Thr399Ile polymorphism there were 7.2% heterozygotes vs 6.2% controls. In most cases, the linkage disequilibrium between the minor alleles Gly299 and Ile399 was confirmed. Increased frequency of both heterozygous genotypes was observed in patients with retinopathy (11.2% for the Asp299Gly). The frequency of the G allele was significantly higher in patients with early onset retinopathy (n = 80) vs patients without DR (odds ratio = 5.0, and 95% confidence interval = 2.33-10.71). In contrast, in the entire retinopathy group, the odds ratio for the G allele was 1.88 (95% confidence interval = 0.93-3.79). In the multivariate logistic regression analysis, the G allele of Asp299Gly was an independent risk factor of early onset DR (p < 0.001). In conclusion, our results suggest an association between the Asp299Gly polymorphism of the TLR4 gene and early onset of DR in the DM2 patients. Thus the G allele may be a predictor of increased risk of retinopathy. PMID- 19135116 TI - A recombinant vesicular stomatitis virus replicon vaccine protects chickens from highly pathogenic avian influenza virus (H7N1). AB - Highly pathogenic avian influenza viruses (HPAIV) of subtypes H5 and H7 cause fatal disease in poultry (fowl plague) but also have zoonotic potential. Currently commercially available vaccines often do not provide sufficient protection and do not allow easy discrimination between vaccinated and infected birds. Therefore, vaccination of domestic poultry against H5 and H7 HPAIV is not allowed in many countries, or is only possible after special permission has been provided. We generated a recombinant marker vaccine based on non-transmissible vesicular stomatitis virus (VSV) expressing the HA antigen of HPAIV A/FPV/Rostock/34 (H7N1) in place of the VSV G gene. This virus, VSV*DeltaG(HA), was propagated on a helper cell line providing VSV G in trans. Since no progeny virus was produced after infection of non-complementing cells, the vector was classified as biosafety level 1 organism ("safe"). Chickens were immunized via the intramuscular route. Following booster vaccination with the same replicons high titers of serum antibodies were induced, which neutralized avian influenza viruses of subtypes H7N1 and H7N7 but not H5N2. Vaccinated chickens were protected against a lethal dose of heterologous HPAIV A/chicken/Italy/445/99 (H7N1). Secretion of challenge virus was short-term and significantly reduced. Finally, it was possible to discriminate vaccinated chickens from infected ones by a simple ELISA assay. We propose that VSV replicons have the potential to be developed to high-quality vaccines for protection of poultry against different subtypes of avian influenza viruses. PMID- 19135115 TI - Long-term vaccine protection from AIDS and clearance of viral DNA following SHIV89.6P challenge. AB - In an earlier study, our group vaccinated rhesus macaques with vesicular stomatitis virus (VSV) vectors expressing Gag, Pol, and Env proteins from a hybrid simian/human immunodeficiency virus (SHIV). This was followed by a single boost with modified vaccinia virus Ankara (MVA) vectors expressing the same proteins. Following challenge with SHIV89.6P, vaccinated animals cleared challenge virus RNA from the blood by day 150 and maintained normal CD4 T cell counts for 8 months. Here we report on the long-term (>5-year post-challenge) status of these animals and the immunological correlates of long-term protection. Using real-time PCR, we found that viral DNA in peripheral blood mononuclear cells (PBMCs) of the vaccinees declined continuously and fell to below detection (<5copies/10(5)cells) by approximately 3 years post-challenge. SHIV DNA was also below the limit of detection in the lymph nodes of two of the four animals at 5 years post-challenge. We detected long-term persistence of multi-functional Gag specific CD8(+) T cells in both PBMCs and lymph nodes of the two protected animals with the Mamu A01(+) MHC I allele. All animals also maintained SHIV89.6P neutralizing antibody titers for 5 years. Our results show that this vaccine approach generates solid, long-term control of SHIV infection, and suggest that it is mediated by both cytotoxic T lymphocytes and neutralizing antibody. PMID- 19135117 TI - A multi-valent vaccine approach that elicits broad immunity within an influenza subtype. AB - Vaccines directed toward individual strains of highly variable viruses like influenza lose efficacy when the circulating viruses no longer resemble the vaccine isolate. Historically, inclusion of more than one isolate per subtype of influenza has been limited by the need to include large doses of antigen with typical protein-based vaccine approaches and by concerns that an immunodominant response to one antigen will limit the response to closely related antigens. Here we provide proof of principle demonstrating that a multi-valent vaccine directed against multiple influenza A virus hemagglutinins (HAs) can elicit broad, neutralizing immunity against multiple strains within a single influenza subtype (H3). We employed a DNA vaccine to direct immunity toward the HA component alone, and a live attenuated influenza virus (LAIV) to assess immunity against the whole virus. Delivery of either HA-DNA or LAIV yielded broad protective immunity across multiple antigenic clusters, including heterologous strains, that was similar to the combined immunity of each antigen assessed separately. Priming with HA-DNA followed by an LAIV boost strengthened and broadened the antibody response toward all three H3 HAs. This prime:boost multi-valent approach was thus able to elicit immunity against multiple strains within the H3 subtype without evidence of immune interference between closely related antigens. Although the trivalent vaccine described here is not a universal vaccine, since protection was limited to circulating viruses from about a two-decade period, these data suggest that full protection within a subtype is possible using this approach with multiple antigens from current and predicted future influenza strains. PMID- 19135118 TI - Immunogenicity of newly constructed attenuated vaccinia strain LC16m8Delta that expresses SIV Gag protein. AB - We developed the method to efficiently construct recombinant vaccinia viruses based on LC16m8Delta strain that can replicate in mammalian cells but is still safe in human. Immunization in a prime-boost strategy using DNA and LC16m8Delta expressing SIV Gag elicited 7-30-fold more IFN-gamma-producing T cells in mice than that using DNA and non-replicating vaccinia DIs recombinant strain. As the previous study on the DNA-prime and recombinant DIs-boost anti-SIV vaccine showed protective efficacy in the macaque model [Someya K, Ami Y, Nakasone T, Izumi Y, Matsuo K, Horibata S, et al. Induction of positive cellular and humoral responses by a prime-boost vaccine encoded with simian immunodeficiency virus gag/pol. J Immunol 2006;176(3):1784-95], LC16m8Delta would have potential as a better recombinant viral vector for HIV vaccine. PMID- 19135119 TI - Metabolism of cisplatin to a nephrotoxin. PMID- 19135121 TI - The effects of acrolein on peroxiredoxins, thioredoxins, and thioredoxin reductase in human bronchial epithelial cells. AB - Inhalation is a common form of exposure to acrolein, a toxic reactive volatile aldehyde that is a ubiquitous environmental pollutant. Bronchial epithelial cells would be directly exposed to inhaled acrolein. The thioredoxin (Trx) system is essential for the maintenance of cellular thiol redox balance, and is critical for cell survival. Normally, thioredoxin reductase (TrxR) maintains the cytosolic (Trx1) and mitochondrial (Trx2) thioredoxins in the reduced state, and the thioredoxins keep the peroxiredoxins (Prx) reduced, thereby supporting their peroxidase function. The effects of acrolein on TrxR, Trx and Prx in human bronchial epithelial (BEAS-2B) cells were determined. A 30-min exposure to 5 microM acrolein oxidized both Trx1 and Trx2, although significant effects were noted for Trx1 at even lower acrolein concentrations. The effects on Trx1 and Trx2 could not be reversed by treatment with disulfide reductants. TrxR activity was inhibited 60% and >85% by 2.5 and 5 microM acrolein, respectively. The endogenous electron donor for TrxR, NADPH, could not restore its activity, and activity did not recover in cells during a 4-h acrolein-free period in complete medium. The effects of acrolein on TrxR and Trx therefore extend beyond the duration of exposure. While there was a strong correlation between TrxR inhibition and Trx1 oxidation, the irreversible effects on Trx1 suggest direct effects of acrolein rather than loss of reducing equivalents from TrxR. Trx2 did not become oxidized until > or = 90% of TrxR was inhibited, but irreversible effects on Trx2 also suggest direct effects of acrolein. Prx1 (cytosolic) and Prx3 (mitochondrial) shifted to a largely oxidized state only when >90 and 100% of their respective Trxs were oxidized. Prx oxidation was readily reversed with a disulfide reductant, suggesting that Prx oxidation resulted from lack of reducing equivalents from Trx and not direct reaction with acrolein. The effects of acrolein on the thioredoxin system and peroxiredoxins could have important implications for cell survival, redox-sensitive cell signaling, and tolerance to other oxidant insults. PMID- 19135122 TI - The profound effects of microcystin on cardiac antioxidant enzymes, mitochondrial function and cardiac toxicity in rat. AB - Deaths from microcystin toxication have widely been attributed to hypovolemic shock due to hepatic interstitial hemorrhage, while some recent studies suggest that cardiogenic complication is also involved. So far, information on cardiotoxic effects of MC has been rare and the underlying mechanism is still puzzling. The present study examined toxic effects of microcystins on heart muscle of rats intravenously injected with extracted MC at two doses, 0.16LD(50) (14 microg MC-LReq kg(-1) body weight) and 1LD(50) (87 microg MC-LReq kg(-1) body weight). In the dead rats, both TTC staining and maximum elevations of troponin I levels confirmed myocardial infarction after MC exposure, besides a serious interstitial hemorrhage in liver. In the 1LD(50) dose group, the coincident falls in heart rate and blood pressure were related to mitochondria dysfunction in heart, while increases in creatine kinase and troponin I levels indicated cardiac cell injury. The corresponding pathological alterations were mainly characterized as loss of adherence between cardiac myocytes and swollen or ruptured mitochondria at the ultrastructural level. MC administration at a dose of 1LD(50) not only enhanced activities and up-regulated mRNA transcription levels of antioxidant enzymes, but also increased GSH content. At both doses, level of lipid peroxides increased obviously, suggesting serious oxidative stress in mitochondria. Simultaneously, complex I and III were significantly inhibited, indicating blocks in electron flow along the mitochondrial respiratory chain in heart. In conclusion, the findings of this study implicate a role for MC-induced cardiotoxicity as a potential factor that should be considered when evaluating the mechanisms of death associated with microcystin intoxication in Brazil. PMID- 19135124 TI - Apoptosis induction of oroxylin A in human cervical cancer HeLa cell line in vitro and in vivo. AB - Oroxylin A is a flavonoid that is found in the roots of Scutellaria baicalensis Georgi. Here, we investigated the antitumor effect of oroxylin A in human cervical cancer HeLa cell line in vitro and in vivo. We found that after inoculated with the HeLa cells the mice treated with oroxylin A showed a significant decrease of tumor volumes and tumor weight compared with the control. Meanwhile, the growth inhibition of oroxylin A on HeLa cells were observed by MTT assay and the value of IC(50) was 19.4+/-0.7 microM after treatment for 48h. Upon our previous research, the inhibition by oroxylin A might be through apoptosis. Then apoptosis induced by oroxylin A in HeLa cells was characterized by DAPI staining and Annexin V/PI double staining, and degradation of PARP (poly-ADP ribose polymerase) was both found in HeLa cells and tumor tissue. Next, activation of the caspase cascade for both the extrinsic and intrinsic pathways were demonstrated in vivo and in vitro, including caspase-8, -9 and -3. We also found that the expression of Bcl-2 protein decreased, which leading to an increase of the Bax/Bcl-2 ratio. Our results showed that oroxylin A exhibited strong antitumor effect in HeLa cell line and apoptosis induction involved in it. PMID- 19135123 TI - Adrenaline in pro-oxidant conditions elicits intracellular survival pathways in isolated rat cardiomyocytes. AB - In several pathologic conditions, like cardiac ischemia/reperfusion, the sustained elevation of plasma and interstitial catecholamine levels, namely adrenaline (ADR), and the generation of reactive oxygen species (ROS) are hallmarks. The present work aimed to investigate in cardiomyocytes which intracellular signalling pathways are altered by ADR redox ability. To mimic pathologic conditions, freshly isolated calcium tolerant cardiomyocytes from adult rat were incubated with ADR alone or in the presence of a system capable of generating ROS [(xanthine with xanthine oxidase) (X/XO)]. ADR elicited a pro oxidant signal with generation of reactive species, which was largely magnified by the ROS generating system. However, no change in cardiomyocytes viability was observed. The pro-oxidant signal promoted the translocation to the nucleus of the transcription factors, Heat shock factor-1 (HSF-1) and Nuclear factor-kappaB (NF kappaB). In addition, proteasome activity was compromised in the experimental groups where the generation of reactive species occurred. The decrease in the proteasome activity of the ADR group resulted from its redox sensitivity, since the activity was recovered by adding the ROS scavenger, tiron. Proteasome inhibition seemed to elicit an increase in HSP70 levels. Furthermore, retention of mitochondrial cytochrome c and inhibition of caspase 3 activity were observed by X/XO incubation in presence or absence of ADR. In conclusion, in spite of all the insults inflicted to the cardiomyocytes, they were capable to activate intracellular responses that enabled their survival. These mechanisms, namely the pathways altered by catecholamine proteasome inhibition, should be further characterized, as they could be of relevance in the ischemia preconditioning and the reperfusion injury. PMID- 19135125 TI - Structure and substrate specificity of acetyltransferase ACIAD1637 from Acinetobacter baylyi ADP1. AB - Gene ACIAD1637 from Acinetobacter baylyi ADP1 encodes a 182 amino acid putative antibiotic resistance protein. The structure of this protein (termed acepita) has been solved in space group P(2) to 2.35 A resolution. Acepita belongs to the GCN5 related N-acetyltransferase (GNAT) family, and contains the four sequence motifs conserved among family members. The structure of acepita is compared with that of pita, its homologue from Pseudomonas aeruginosa. Acepita has a similar substrate profile to pita and performs a similar function. PMID- 19135126 TI - Ang-II-induced Ca(2+) influx is mediated by the 1/4/5 subgroup of the transient receptor potential proteins in cultured aortic smooth muscle cells from diabetic Goto-Kakizaki rats. AB - Angiotensin-II (Ang-II) exerts many of its vascular effects, including the pathophysiological changes associated with type 2 diabetes, through changes in intracellular calcium concentration [Ca(2+)](i). We sought to clarify the mechanism responsible for Ang-II-induced Ca(2+) influx in cultured aortic VSMC using the Goto-Kakizaki (GK) rat model of type 2 diabetes. Ang-II-induced Ca(2+) influx was blocked by neither VDCC nor c-src inhibition but was sensitive to inositol 1,4,5-trisphosphate receptor inhibition, lanthanide and the diacylglycerol analogue, oleoyl-2-acetyl-sn-glycerol. Since transient receptor potential canonical (TRPC)-3 gene expression was undetectable in both WKY and GK VSMCs and TRPC6 gene and protein expression were significantly down-regulated in GK, we believe the 1/4/5 subgroup of TRPC proteins plays a significant role. Furthermore, in GK VSMC the elevated calcium influx observed was not attributable to increased TRPC expression, but rather an alteration of TRPC activity. PMID- 19135127 TI - The constitutive activity of the ghrelin receptor attenuates apoptosis via a protein kinase C-dependent pathway. AB - The ghrelin receptor (GHS-R1a) displays a high level of constitutive signaling through a phospholipase C/protein kinase C-dependent pathway. Therefore, we have investigated the role of agonist-dependent and agonist-independent signaling of GHS-R1a in apoptosis using the seabream GHS-R1a stably expressed in human embryonic kidney 293 cells (HEK-sbGHS-R1a cells). Cadmium-induced activation of caspase-3 was significantly attenuated in HEK-sbGHS-R1a cells compared to wild type HEK293 cells, while the apoptotic responses to the protein kinase C inhibitor staurosporine were similar. GHS-R1a ligands had no effect on caspase-3 activation or on cell proliferation. Concentrations of the inverse agonist [d Arg(1),d-Phe(5),d-Trp(7,9),Leu(11)]-substance P sufficient to inhibit constitutive inositol phosphate generation did not enhance caspase-3 activity, suggesting a possible role of phosphatidylcholine-specific phospholipase C in the anti-apoptotic activity of GHS-R1a. In conclusion, our data suggests that the constitutive activity of sbGHS-R1a may be sufficient alone to attenuate apoptosis via a protein kinase C-dependent pathway. PMID- 19135128 TI - Insulin-dependent regulation of GLAST/EAAT1 in Bergmann glial cells. AB - Glutamate is the major excitatory neurotransmitter in the central nervous system. Ionotropic and metabotropic glutamate receptors are present in neurons and glial cells and are involved in gene expression regulation. A family of sodium dependent glutamate transporters carries out the removal of the neurotransmitter from the synaptic cleft. In the cerebellum, the bulk of glutamate transport is mediated through the excitatory amino acids transporter 1 (EAAT1/GLAST) expressed in Bergmann glial cells. Proper transporter function is critical for glutamate cycling and glucose turnover, as well as prevention of excitotoxic insult to Purkinje cells. In order to gain insight into the regulatory signals that modify this uptake activity, we investigated the effects of insulin exposure. Using the well-defined chick cerebellar Bergmann glial cell culture model, we observed a time and dose-dependent decrease in [(3)H]-d-aspartate uptake. As expected, this effect is mimicked by the tyrosine phosphatase inhibitor sodium orthovanadate, suggesting a receptor-mediated effect. Equilibrium [(3)H]-d-aspartate binding experiments as well as a reverse transcriptase/polymerase chain reaction strategy demonstrated that the decrease in the uptake activity is related to reduced numbers of transporter molecules in the plasma membrane. Accordingly, the transcriptional activity of the chick glast promoter diminished upon insulin treatment. The present findings suggest the involvement of insulin in neuronal/glial coupling in the cerebellum. PMID- 19135129 TI - Nuclear size of c-Fos expression at the auditory brainstem is related to the time varying nature of the acoustic stimuli. AB - Acoustic stimulation is known to induce c-Fos expression in the auditory system but how the expression might be related to the time-variance of the sound (e.g., steady or frequency-varying) is unclear. Here we measured morphometrically Fos immunohistochemical stains at the auditory brainstem after exposing rats to a pure tone or a narrow-range frequency modulated (FM) sound for various durations (10, 30 or 90 min). Nuclear sizes of Fos-stains at the cochlear nucleus (CN) and inferior colliculus (IC) were estimated under microscope. Tone stimulation at the given frequency (9 kHz) induced Fos-expression at locations consistent with the tonotopic maps, and no clear difference in the spatial distribution of the Fos stains was observed across stimuli. In general, more Fos-stains appeared after longer stimulations and most notably cell nuclei labeled with Fos immunoreactivity were statistically larger in size under longer pure tone and FM tone stimulations. Findings suggest that Fos-expression mechanisms are related to the selective response of different subpopulations of neurons to sounds of different time-varying properties. Results support that the time-variance of acoustic stimulation should be considered in the interpretation of Fos-expression findings. PMID- 19135130 TI - Nortriptyline mediates behavioral effects without affecting hippocampal cytogenesis in a genetic rat depression model. AB - A prevailing hypothesis is that neurogenesis is reduced in depression and that the common mechanism for antidepressant treatments is to increase it in adult hippocampus. Reduced neurogenesis has been shown in healthy rats exposed to stress, but it has not yet been demonstrated in depressed patients. Emerging studies now indicate that selective serotonin reuptake inhibitors can, exert behavioral effects without affecting neurogenesis in mice. Here we extend our previous findings demonstrating that the number of BrdU positive cells in hippocampus was significantly higher in a rat model of depression, the Flinders Sensitive Line (FSL) compared to the control strain the Flinders Resistant Line (FRL). We also show that chronic treatment with the tricyclic antidepressant nortriptyline exerts behavioral effects in the Porsolt forced swim test without affecting hippocampal cell proliferation in the FSL model. These results strengthen the arguments against hypothesis of neurogenesis being necessary in etiology of depression and as requisite for effects of antidepressants, and illustrate the importance of using a disease model and not healthy animals to assess effects of potential therapies for major depressive disorder. PMID- 19135131 TI - The molecular mechanism of dexamethasone-mediated effect on the blood-brain tumor barrier permeability in a rat brain tumor model. AB - This study was performed to determine whether dexamethasone (DEX) had an effect on calcium-activated potassium channels (K(Ca) channels) and Occludin protein in blood-brain tumor barrier (BTB). Using a rat brain glioma model, we found that the expression of K(Ca) channels protein and Occludin protein was significantly increased in brain tumor tissue after DEX treatment for 3 days. Compared with non DEX-treated animals, Evans Blue levels were greatly attenuated in DEX-treated animals. These effects were significantly reversed by the glucocorticoid receptor antagonist RU38486. In addition, DEX treatment enhanced the density of I(KCa) in the rat brain microvascular endothelial cells (RBMECs) in vitro BTB. All of these results strongly suggest that DEX could be involved in the regulation of both transcellular and paracellular pathway. PMID- 19135132 TI - Resveratrol inhibits MMP-9 expression by up-regulating PPAR alpha expression in an oxygen glucose deprivation-exposed neuron model. AB - Resveratrol (trans-3,4',5-trihydroxystilbene, Res) is a natural polyphenol. A recent experiment confirmed that Res can selectively activate both peroxisome proliferators-activated receptors (PPAR) alpha and gamma. In addition, Res can protect neurons by matrix metalloproteinase-9 (MMP-9) down-regulation. The relationship between Res, MMP-9 and PPAR alpha or gamma was studied in an oxygen glucose deprivation-exposed neuron model. It showed that Res can inhibit mRNA and protein expression of MMP-9, while it up-regulates the expression of PPAR alpha and gamma. The effect of Res on both PPAR alpha and MMP-9 can be offset partially by MK886. However, PPAR gamma antagonist GW9662 had little effect on MMP-9 expression. These results suggest that Res can inhibit MMP-9 expression by up regulating PPAR alpha. PMID- 19135133 TI - Cognitive performance and age-related changes in the hippocampal proteome. AB - Declining cognitive performance is associated with increasing age, even in the absence of overt pathological processes. We and others have reported that declining cognitive performance is associated with age-related changes in brain glucose utilization, long-term potentiation and paired-pulse facilitation, protein expression, neurotransmitter levels, and trophic factors. However, it is unclear whether these changes are causes or symptoms of the underlying alterations in dendritic and synaptic morphology that occur with age. In this study, we examined the hippocampal proteome for age- and cognition-associated changes in behaviorally stratified young and old rats, using two-dimensional in gel electrophoresis and MS/MS. Comparison of old cognitively intact with old cognitively impaired animals revealed additional changes that would not have been detected otherwise. Interestingly, not all age-related changes in protein expression were associated with cognitive decline, and distinct differences in protein expression were found when comparing old cognitively intact with old cognitively impaired rats. A large number of protein changes with age were related to the glycolysis/gluconeogenesis pathway. In total, the proteomic changes suggest that age-related alterations act synergistically with other perturbations to result in cognitive decline. This study also demonstrates the importance of examining behaviorally-defined animals in proteomic studies, as comparison of young to old animals regardless of behavioral performance would have failed to detect many cognitive impairment-specific protein expression changes evident when behavioral stratification data were used. PMID- 19135135 TI - Increased impulsive behavior and risk proneness following lentivirus-mediated dopamine transporter over-expression in rats' nucleus accumbens. AB - Multiple theories have been proposed for sensation seeking and vulnerability to impulse-control disorders [Zuckerman M, Kuhlman DM (2000) Personality and risk taking: Common biosocial factors. J Pers 68:999-1029], and many of these rely on a dopamine system deficit. Available animal models reproduce only some behavioral symptoms and seem devoid of construct validity. We used lentivirus tools for over expressing or silencing the dopamine transporter (DAT) and we evaluated the resulting behavioral profiles in terms of motivation and self-control. Wistar adult rats received stereotaxic inoculation of a lentivirus that allowed localized intra-accumbens delivery of a DAT gene enhancer/silencer, or the green fluorescent protein, GFP. These animals were studied for intolerance to delay, risk proneness and novelty seeking. As expected, controls shifted their demanding from a large reward toward a small one when the delivery of the former was increasingly delayed (or uncertain). Interestingly, in the absence of general locomotor effects, DAT over-expressing rats showed increased impulsivity (i.e. a more marked shift of demanding from the large/delayed toward the small/soon reward), and increased risk proneness (i.e. a less marked shift from the large/uncertain toward the small/sure reward), compared with controls. Rats with enhanced or silenced DAT expression did not show any significant preference for a novel environment. In summary, consistent with literature on comorbidity between attention-deficit/hyperactivity disorder and pathological gambling, we demonstrate that DAT over-expression in rats' nucleus accumbens leads to impulsive and risk prone phenotype. Thus, a reduced dopaminergic tone following altered accumbal DAT function may subserve a sensation-seeker phenotype and the vulnerability to impulse-control disorders. PMID- 19135134 TI - 5-HT inhibition of rat insulin 2 promoter Cre recombinase transgene and proopiomelanocortin neuron excitability in the mouse arcuate nucleus. AB - A number of anti-obesity agents have been developed that enhance hypothalamic 5 HT transmission. Various studies have demonstrated that arcuate neurons, which express proopiomelanocortin peptides (POMC neurons), and neuropeptide Y with agouti-related protein (NPY/AgRP) neurons, are components of the hypothalamic circuits responsible for energy homeostasis. An additional arcuate neuron population, rat insulin 2 promoter Cre recombinase transgene (RIPCre) neurons, has recently been implicated in hypothalamic melanocortin circuits involved in energy balance. It is currently unclear how 5-HT modifies neuron excitability in these local arcuate neuronal circuits. We show that 5-HT alters the excitability of the majority of mouse arcuate RIPCre neurons, by either hyperpolarization and inhibition or depolarization and excitation. RIPCre neurons sensitive to 5-HT, predominantly exhibit hyperpolarization and pharmacological studies indicate that inhibition of neuronal firing is likely to be through 5-HT(1F) receptors increasing current through a voltage-dependent potassium conductance. Indeed, 5 HT(1F) receptor immunoreactivity co-localizes with RIPCre green fluorescent protein expression. A minority population of POMC neurons also respond to 5-HT by hyperpolarization, and this appears to be mediated by the same receptor-channel mechanism. As neither POMC nor RIPCre neuronal populations display a common electrical response to 5-HT, this may indicate that sub-divisions of POMC and RIPCre neurons exist, perhaps serving different outputs. PMID- 19135136 TI - c-Jun N-terminal kinase binding domain-dependent phosphorylation of mitogen activated protein kinase kinase 4 and mitogen-activated protein kinase kinase 7 and balancing cross-talk between c-Jun N-terminal kinase and extracellular signal regulated kinase pathways in cortical neurons. AB - The c-Jun N-terminal kinase (JNK) is a mitogen-activated protein kinase (MAPK) activated by stress-signals and involved in many different diseases. Previous results proved the powerful effect of the cell permeable peptide inhibitor d JNKI1 (d-retro-inverso form of c-Jun N-terminal kinase-inhibitor) against neuronal death in CNS diseases, but the precise features of this neuroprotection remain unclear. We here performed cell-free and in vitro experiments for a deeper characterization of d-JNKI1 features in physiological conditions. This peptide works by preventing JNK interaction with its c-Jun N-terminal kinase-binding domain (JBD) dependent targets. We here focused on the two JNK upstream MAPKKs, mitogen-activated protein kinase kinase 4 (MKK4) and mitogen-activated protein kinase kinase 7 (MKK7), because they contain a JBD homology domain. We proved that d-JNKI1 prevents MKK4 and MKK7 activity in cell-free and in vitro experiments: these MAPKK could be considered not only activators but also substrates of JNK. This means that d-JNKI1 can interrupt downstream but also upstream events along the JNK cascade, highlighting a new remarkable feature of this peptide. We also showed the lack of any direct effect of the peptide on p38, MEK1, and extracellular signal-regulated kinase (ERK) in cell free, while in rat primary cortical neurons JNK inhibition activates the MEK1-ERK-Ets1/c-Fos cascade. JNK inhibition induces a compensatory effect and leads to ERK activation via MEK1, resulting in an activation of the survival pathway-(MEK1/ERK) as a consequence of the death pathway-(JNK) inhibition. This study should hold as an important step to clarify the strong neuroprotective effect of d-JNKI1. PMID- 19135137 TI - Whole-body periodic acceleration reduces brain damage in a focal ischemia model. AB - Stroke is the second most common cause of death and major cause of disability worldwide. Actual treatment involves surgery and/or thrombolytic drugs, but there is an urgent need for new approaches. Periodic acceleration, a rocking headward to footward movement of the whole body, is a non-invasive method to induce pulsatile shear stress on the vascular endothelium eliciting an enhanced production and secretion of endothelium-derived products such as nitric oxide, prostacyclin, prostaglandin E2, tissue plasminogen activator (tPA), and adrenomedullin. All these products have been shown to protect the brain from ischemic injuries. A rat model of focal brain ischemia was treated with application of periodic acceleration for 3 h immediately after the onset of ischemia. Controls remained static for the same period of time. Brain damage was assessed by magnetic resonance imaging (MRI) and biochemical markers. A significant reduction in brain damage was observed, 7 days post-ischemia, in rocked rats when compared with the static controls, through MRI. Furthermore, rocked animals had significantly lower levels of Beclin 1 and fractin than their static counterparts, and some isoforms of nitric oxide synthase were regulated by periodic acceleration. Our results show that periodic acceleration may provide a novel, affordable, non-invasive therapeutic option for the treatment of stroke. PMID- 19135138 TI - Decaarginine-PEG-liposome enhanced transfection efficiency and function of arginine length and PEG. AB - Oligoarginine-conjugated lipids ((Arg)n-PEG-lipid) (n=4, 6, 8, and 10: number of arginine residues) are novel gene delivery vectors. We prepared two oligoarginine modified liposomes using (Arg)n-lipid without and with poly(ethylene glycol) (PEG) spacer ((Arg)n-L and (Arg)n-PEG-L), and investigated the effect of PEG spacer and oligoarginine length of liposomes on cellular uptake, gene transfection, and its mechanism in HeLa cells, using complexes with plasmid DNA (DNA) or oligodeoxynucleotide. Transfection efficiency increased as the number of arginine residues increased and Arg10-PEG-L/DNA complexes (lipoplexes) showed the highest gene transfection efficiency among (Arg)n- and (Arg)n-PEG-lipoplexes. Arg4- and Arg4-PEG-lipoplexes were taken up greatly into cells, but showed lower transfection efficiency than Arg10- and Arg10-PEG-lipoplexes, respectively. The different gene expression by Arg4-L to Arg10-L with or without PEG spacer may be explained by the different intracellular uptake mechanism. The main cellular uptake mechanism of Arg10-L and Arg10-PEG-L was the macropinocytosis pathway, whereas that of Arg4-L and Arg4-PEG-L was not. PEG spacer was more effective for intracellular trafficking than Arg length and surface charge of lipoplex which depends on Arg length at the almost same size of lipoplexes. The findings suggested that Arg10-PEG-L was a superior vector since Arg10 induced the macropinocytosis uptake pathway. PMID- 19135139 TI - Chitosan N-betainates/DNA self-assembly nanoparticles for gene delivery: in vitro uptake and transfection efficiency. AB - The aim of this work is to investigate the effect of betaine substitution degree of chitosan N-betainates (CsB) on cellular uptake, cytotoxicity and transfection efficiency of CsB/DNA complex nanoparticles (CsBNs) against COS-7 and MDA-MB-468 cells. The polymers with three substitution degrees (CsB12, CsB47 and CsB85) complexed with pDNA formed CsBN12s, CsBN47s and CsBN85s. The CsBNs showed less pH dependency with smaller particle size and higher zeta potential than that of chitosan/pDNA complex nanoparticles (CsNs) at neutral pH. CsBN85s showed stronger cellular uptake than that of CsBN47s or CsBN12s. CsBNs showed higher cytotoxicity than CsNs, and a trend increasing toxicity with substitution degree increasing. In COS-7 cells, the transfection efficiency increased with the substitution degree increasing, while the opposite result was observed in MDA-MB-468 cells. Chitosan modified with betaine could increase its ability to facilitate DNA uptake and its cytotoxicity, both of which showed the influence on transfection efficiency. It was able to increase cellular uptake and transfection efficiency of complex nanoparticles in COS-7 cells to increase betaine substitution of CsB, however, the higher sensitivity of MDA-MB-468 cells to CsBs led to decreased transfection efficiency due to the increased cytotoxicity with betaine substitution increasing. The predominant role of cellular uptake or toxicity in affecting transfection efficiency was different in two cell lines. These results provided an important guidepost for further development of chitosan derivatives/pDNA complexes as non-viral gene vectors. PMID- 19135140 TI - The protection of selenium on ROS mediated-apoptosis by mitochondria dysfunction in cadmium-induced LLC-PK(1) cells. AB - Selenium, an essential trace element, showed the significant protective effects against liver and kidney damage induced by some heavy metals. However, the mechanism how selenium suppresses cadmium (Cd)-induced cytotoxicity remains unclear. In this study, we investigated the protective mechanism of selenium on Cd-induced apoptosis in LLC-PK(1) cells via reactive oxygen species (ROS) and mitochondria linked signal pathway. Studies of PI and Annexin V dual staining analysis demonstrated that 20 microM Cd-induced apoptosis as early as 18 h. A concomitant by the generation of ROS, the loss of mitochondrial membrane potential, cytochrome c (cyt c) release, activation of caspase-9, -3 and regulation of Bcl-2 and Bax were observed. N-acetylcysteine (NAC, 500 microM), a free radical scavenger, was used to determine the involvement of ROS in Cd induced apoptosis. During the process, selenium played the same role as NAC. The anti-apoptosis exerted by selenium involved the blocking of Cd-induced ROS generation, the inhibition of Cd-induced mitochondrial membrane potential collapse, the prevention of cyt c release, subsequent inhibition of caspase activation and the changed level of Bcl-2 and Bax. Taken together, we concluded that Cd-induced apoptosis was mediated by oxidative stress and selenium produced a significant protection against Cd-induced apoptosis in LLC-PK(1) via ameliorating the mitochondrial dysfunction. PMID- 19135141 TI - Potential method to determine irritant potency in vitro - Comparison of two reconstructed epidermal culture models with different barrier competency. AB - Testing chemicals for their ability to cause skin irritation is required for all ingredients of products that come into contact with the skin. Here, we describe a potential method for determining the irritant potency of a chemical in vitro and apply the method to two different reconstructed epidermis models which exhibit different barrier properties. Two surfactants: sodium dodecyl sulphate, Triton X100 and two non-surfactants: 2-4-di-nitro-chloro-benzene, cinnamaldehyde were applied topically in a dose response for 24h. Biomarkers IL-1alpha, IL-1RA, IL-8 and MTT were assessed and EC(50) values determined. Variation in barrier properties between the epidermal models led to variation in the extent of penetration of surfactants, but not of non-surfactants which in turn influenced the EC(50) value obtained from surfactants. Furthermore, EC(50) values showed that no single biomarker could be classed as the most sensitive biomarker since biomarker sensitivity differed between the different chemicals studied. However, the ranking of the chemicals in order of strong to weak irritant was the same irrespective of the model used and also independent of the biomarker used (Triton X100>DNCB>SDS>CA). This study describes a method which not only distinguishes an irritant from a non-irritant but which may possibly also be used to determine irritant potency. PMID- 19135142 TI - New strategy for alerting central nervous system toxicity: Integration of blood brain barrier toxicity and permeability in neurotoxicity assessment. AB - The combination of an in vitro BBB model (4d/24w) with a neuronal cell line (SH SY5Y) provides a convenient approach to explore the importance of BBB permeability in neurotoxicity assessment of compounds. The toxicity of 16 compounds on SH-SY5Y cells was evaluated after 24h incubation with each compound and compared to their toxicity on SH-SY5Y after passage through the BBB model. Nine out of 16 compounds were found toxic after direct exposure at 100muM while only three still induced toxicity on SH-SY5Y cells after BBB transport. The BBB permeability values of each compound revealed that in the case of compounds that did not induce toxicity, the amount that crossed the BBB was not enough to exert a toxic effect on the neuronal cells. Since disrupting the BBB may also cause unwanted effect on brain cells, the BBB toxicity of these compounds have been assessed. Our results prompted the importance of BBB permeability assessment in neurotoxicity evaluation, as it allows a better estimation of the actual concentration at the target site. PMID- 19135143 TI - Inhibitory vs. protective effects of N-acetyl-l-cysteine (NAC) on the electromechanical properties of the spontaneously beating atria of the frog (Rana ridibunda): an ex vivo study. AB - The results of this study have shown that N-acetyl-l-cysteine (NAC), a compound used for protection of tissues or cell cultures against the deleterious effects of various environmental pollutants, has certain unusual effects on the contraction of the spontaneously beating atria of the frog isolated in saline (ex vivo): (1) NAC, 6.0 and 10.0mM, eliminated, in a concentration-dependent manner, the contractile properties of the atria (force and frequency) within minutes, without affecting its electrical properties; (2) the IC(50) of NAC for the force was 5.09+/-1.01 mM (n=6) [4.98-5.19 mM, 95% confidence interval (CI)], significantly lower than the IC(50) for the frequency, 6.15+/-1.01 mM, (6.02-6.29 mM, 95% CI), indicating that working atria cells are more sensitive to NAC than autorhythmic cells. The no-observed-effect concentration (NOEC) was 1-2mM; (3) the pattern of NAC-induced inhibition of electromechanical activity was similar to that of verapamil, an indication that NAC possibly affects L-type voltage gated calcium channels; (4) NAC at 2mM protected against cadmium-induced inhibition of atria contraction. The IC(50) for cadmium was 17.9+/-1.1 microM (n=6) (16.9-19.0 microM, 95% CI), while in the presence of 2mM NAC, it became 123.3+/-1.0 microM (n=6) (114.8-132.4 microM, 95% CI). The same concentration of NAC failed to exert any protective effects against rotenone (5 microM)-induced inhibition of atria contraction. The protective effects of NAC are probably due to chelation of cadmium, rather than scavenging of oxidants. PMID- 19135144 TI - Phylogenetic analysis of the eukaryotic RNA (cytosine-5)-methyltransferases. AB - RNA (cytosine-5)-methyltransferases (RCMTs) have been characterized both in prokaryotic and eukaryotic organisms. The RCMT family, however, remains largely uncharacterized, as opposed to the family of DNA (cytosine-5)-methyltransferases which has been studied in depth. In the present study, an in silico identification of the putative 5-methylcytosine RNA-generating enzymes in the eukaryotic genomes was performed. A comprehensive phylogenetic analysis of the putative eukaryotic RCMT-related proteins has been performed in order to redefine subfamilies within the RCMT family. Five distinct eukaryotic subfamilies were identified, including the three already known (NOP2, NCL1 and YNL022c), one novel subfamily (RCMT9) and a fifth one which hitherto was considered to exist exclusively in prokaryotes (Fmu). The potential evolutionary relationships among the different eukaryotic RCMT subfamilies were also investigated. Furthermore, the results of this study add further support to a previous hypothesis that RCMTs represent evolutionary intermediates of RNA (uridine-5)-methyltransferases and DNA (cytosine-5)-methyltransferases. PMID- 19135145 TI - Consequences of different housing conditions on brain morphology in laying hens. AB - The aim of this study was to analyze the impact of physical and social stress on the avian forebrain morphology. Therefore, we used laying hens kept in different housing systems from puberty (approximately 16 weeks old) until the age of 48 weeks: battery cages, small littered ground pen, and free range system. Cell body sizes and catecholaminergic and serotonergic innervation patterns were investigated in brain areas expected to be sensitive to differences in environmental stimulation: hippocampal substructures and the nidopallium caudolaterale (NCL), a functional analogue of the prefrontal cortex. Our analysis shows both structures differing in the affected morphological parameters. Compared to battery cage hens, hens in the free range system developed larger cells in the dorsomedial hippocampus. Only these animals exhibited an asymmetry in the tyrosine hydroxylase density with more fibres in the left dorsomedial hippocampus. We assume that the higher spatial complexity of the free range system is the driving force of these changes. In contrast, in the NCL the housing systems affected only the serotonergic innervation pattern with highest fibre densities in free range hens. Moreover hens of the free range system displayed the worst plumage condition, which most likely is caused by feather pecking causing an altered serotonergic innervation pattern. Considering the remarkable differences between the three housing conditions, their effects on hippocampal structures and the NCL were surprisingly mild. This observation suggests that the adult brain of laying hens displays limited sensitivity to differences in social and physical environment induced post-puberty, which warrants further studies. PMID- 19135146 TI - Superoxide dismutase is regulated by LAMMER kinase in Drosophila and human cells. AB - LAMMER kinases (also known as CDC-2-like or CLKs) are a family of dual specificity serine/threonine protein kinases that are found in all sequenced eukaryotic genomes. In the fission yeast, Schizosaccharomyces pombe, the LAMMER kinase gene, Lkh1, positively regulates the expression of the antioxidant defense genes, superoxide dismutase 1 (sod1+, CuZn-SOD) and catalase (ctt1+, CAT). We have shown that mutations in the Drosophila LAMMER kinase gene, Darkener of apricot (Doa), protect against the decrease in life span caused by the reactive oxygen species (ROS) generator paraquat, and at the same time show an increase in cytoplasmic (CuZn-Sod or SOD1) and mitochondrial superoxide dismutase (Mn-Sod or SOD2) protein levels and activity. The siRNA-mediated knock down of the human LAMMER kinase gene, CLK-1, in HeLa and MCF-7 human cell lines leads to an increase in both SOD1 activity and mRNA transcript levels. These data suggest that SOD1 is negatively regulated by LAMMER kinases in Drosophila and human cell lines and that this regulation may be conserved during evolution. PMID- 19135147 TI - 4-Hydroxynonenal enhances CD36 expression on murine macrophages via p38 MAPK mediated activation of 5-lipoxygenase. AB - Increased levels of 4-hydroxynonenal (HNE) and 5-lipoxygenase (5-LO) coexist in atherosclerotic lesions but their relationship in atherogenesis is unclear. This study investigated the role of 5-LO in HNE-induced CD36 expression and macrophage foam cell formation, and the link between HNE and 5-LO. In J774A.1 murine macrophages, HNE (10 microM) enhanced CD36 expression in association with an increased uptake of oxLDL, which was blunted by inhibition of 5-LO with MK886, a 5-LO inhibitor, or with 5-LO siRNA. In peritoneal macrophages from 5-LO-deficient mice, HNE-induced CD36 expression was markedly attenuated, confirming a pivotal role of 5-LO in HNE-induced CD36 expression. In an assay for 5-LO activity, stimulation of macrophages with HNE led to increased leukotriene B(4) production in the presence of exogenous arachidonic acid in association with an increased association of 5-LO to the nuclear membrane. Among the mitogen-activated protein kinase (MAPK) pathways involved in 5-LO phosphorylation, HNE predominantly activated p38 MAPK in macrophages, and the p38 MAPK inhibitor SB203580, but not an extracellular signal-regulated kinase inhibitor, suppressed HNE-induced LTB(4) production. Collectively, these data suggest that p38 MAPK-mediated activation of 5-LO by HNE might enhance CD36 expression, consequently leading to the formation of macrophage foam cells. PMID- 19135149 TI - (Pre)diabetes, brain aging, and cognition. AB - Cognitive dysfunction and dementia have recently been proven to be common (and underrecognized) complications of diabetes mellitus (DM). In fact, several studies have evidenced that phenotypes associated with obesity and/or alterations on insulin homeostasis are at increased risk for developing cognitive decline and dementia, including not only vascular dementia, but also Alzheimer's disease (AD). These phenotypes include prediabetes, diabetes, and the metabolic syndrome. Both types 1 and 2 diabetes are also important risk factors for decreased performance in several neuropsychological functions. Chronic hyperglycemia and hyperinsulinemia primarily stimulates the formation of Advanced Glucose Endproducts (AGEs), which leads to an overproduction of Reactive Oxygen Species (ROS). Protein glycation and increased oxidative stress are the two main mechanisms involved in biological aging, both being also probably related to the etiopathogeny of AD. AD patients were found to have lower than normal cerebrospinal fluid levels of insulin. Besides its traditional glucoregulatory importance, insulin has significant neurothrophic properties in the brain. How can clinical hyperinsulinism be a risk factor for AD whereas lab experiments evidence insulin to be an important neurothrophic factor? These two apparent paradoxal findings may be reconciliated by evoking the concept of insulin resistance. Whereas insulin is clearly neurothrophic at moderate concentrations, too much insulin in the brain may be associated with reduced amyloid-beta (Abeta) clearance due to competition for their common and main depurative mechanism - the Insulin-Degrading Enzyme (IDE). Since IDE is much more selective for insulin than for Abeta, brain hyperinsulinism may deprive Abeta of its main clearance mechanism. Hyperglycemia and hyperinsulinemia seems to accelerate brain aging also by inducing tau hyperphosphorylation and amyloid oligomerization, as well as by leading to widespread brain microangiopathy. In fact, diabetes subjects are more prone to develop extense and earlier-than-usual leukoaraiosis (White Matter High-Intensity Lesions - WMHL). WMHL are usually present at different degrees in brain scans of elderly people. People with more advanced WMHL are at increased risk for executive dysfunction, cognitive impairment and dementia. Clinical phenotypes associated with insulin resistance possibly represent true clinical models for brain and systemic aging. PMID- 19135148 TI - Glucose-mediated tyrosine nitration in adipocytes: targets and consequences. AB - Hyperglycemia, a key factor in insulin resistance and diabetic pathology, is associated with cellular oxidative stress that promotes oxidative protein modifications. We report that protein nitration is responsive to changes in glucose concentrations in 3T3-L1 adipocytes. Alterations in the extent of tyrosine nitration as well as the cellular nitroproteome profile correlated tightly with changing glucose concentrations. The target proteins we identified are involved in fatty acid binding, cell signaling, protein folding, energy metabolism, antioxidant capacity, and membrane permeability. The nitration of adipocyte fatty acid binding protein (FABP4) at Tyr19 decreases, similar to phosphorylation, the binding of palmitic acid to the fatty acid-free protein. This potentially alters intracellular fatty acid transport, nuclear translocation of FABP4, and agonism of PPAR gamma. Our results suggest that protein tyrosine nitration may be a factor in obesity, insulin resistance, and the pathogenesis of diabetes. PMID- 19135150 TI - Insights into a putative hinge region in elastin using molecular dynamics simulations. AB - The resiliency and elasticity of vertebrate tissues are traced to elastin, a crosslinked protein with extensive hydrophobic regions. There is little discussion in the literature on the structure and dynamics of the alanine-rich crosslinking regions of elastin that comprise a significant part of the native protein. In particular, the region encoded by exons 21 and 23, a contiguous splice form found in all types of human elastin, is believed to be strategically positioned for proper function of the protein, namely, in the reversible elongation and contraction of tissue. Hence, molecular dynamics (MD) calculations on the EX21/23 domain are reported here. This crosslinking domain has been assumed to adopt an architecture in which the putative hinge region links two alpha-helices. In this paper, we use a homology-based approach to obtain starting structures in the hinge region. The subsequent MD brings new insights into the possibility of fluctuations between "open" and "closed" states, as well as distinguishing structural features of the latter. The significance of these findings towards an enhanced understanding of structure-function relationships in elastin and the elastic fiber is discussed. PMID- 19135151 TI - h channel-dependent deficit of theta oscillation resonance and phase shift in temporal lobe epilepsy. AB - I(h) tunes hippocampal CA1 pyramidal cell dendrites to optimally respond to theta inputs (4-12 Hz), and provides a negative time delay to theta inputs. Decreased I(h) activity, as seen in experimental temporal lobe epilepsy (TLE), could significantly alter the response of dendrites to theta inputs. Here we report a progressive erosion of theta resonance and phase lead in pyramidal cell dendrites during epileptogenesis in a rat model of TLE. These alterations were due to decreased I(h) availability, via a decline in HCN1/HCN2 subunit expression resulting in decreased h currents, and altered kinetics of the residual channels. This acquired HCN channelopathy thus compromises temporal coding and tuning to theta inputs in pyramidal cell dendrites. Decreased theta resonance in vitro also correlated with a reduction in theta frequency and power in vivo. We suggest that the neuronal/circuitry changes associated with TLE, including altered I(h) dependent inductive mechanisms, can disrupt hippocampal theta function. PMID- 19135152 TI - GABA(B) presynaptic receptors modulate glycine exocytosis from mouse spinal cord and hippocampus glycinergic nerve endings. AB - Glycine release and modulation of glycine exocytosis by presynaptic receptors have been rarely studied. We here investigate on the existence and the pharmacological profile of GABA(B) receptors regulating glycine release in mouse spinal cord and hippocampus. Synaptosomes were preincubated with [(3)H]glycine in presence of the glycine transporter 1 inhibitor NFPS, in order to study release from glycinergic terminals selectively labelled through the glycine transporter 2, and depolarized in superfusion with concentrations of KCl (12 and 15 mM) previously found to elicit exocytosis of glycine. Exposure of synaptosomes to (-) baclofen caused inhibition of [(3)H]glycine overflow: EC(50)=0.62+/-0.08 microM, E(max) approximately 65% in the spinal cord and EC(50)=0.58+/-0.07 microM, E(max) approximately 55% in the hippocampus. The effect of 3 microM (-)-baclofen in the spinal cord was prevented by the GABA(B) receptor antagonists CGP 52432 (IC(50)=22.6+/-2.15 microM) and CGP 35348 (IC(50)=5.99+/-1.21 microM), whereas phaclofen was ineffective. In the hippocampus, the CGP antagonists were more potent than in the spinal cord, their IC(50) values amounting to 0.81+/-0.08 microM and to 0.94+/-0.09 microM, respectively; phaclofen (100-300 microM) was a weak antagonist. It is concluded that: (i) glycinergic nerve terminals in spinal cord and hippocampus possess GABA(B) receptors mediating inhibition of the evoked glycine exocytosis; (ii) the pharmacological profiles of the receptors are qualitatively similar in the two regions; (iii) low concentrations of GABA(B) antagonists appear able to preferentially inhibit the receptors located on hippocampal glycinergic nerve endings. PMID- 19135154 TI - The personality trait openness is related to cerebral 5-HTT levels. AB - Potentiation of serotonergic transmission increases cognitive flexibility, but can in other circumstances increase sensitivity to stressful environmental cues. The personality trait Openness to Experience reflects and is also associated with an increased risk for mood disorders. We hypothesized that the personality trait has an association with a biomarker of serotonergic transmission, the plasma membrane serotonin transporter (5-HTT). In 50 healthy volunteers, we tested for correlations between scores on the NEO-PI-R scale Openness to Experience and its subscales, and cerebral binding of the 5-HTT selective PET radioligand [11C]DASB. Subjects were genotyped for the 5-HTT long/short polymorphism, and for a single nucleotide polymorphism in the long allele, designated LA/LG. Midbrain [11C]DASB binding correlated negatively with scores for Openness to Experience and its two subscales, Openness to Actions and Openness to Values. The latter subscore was negatively correlated with [11C]DASB binding in all brain regions in which [11C]DASB binding was quantified. Genetic analysis showed that homozygote LA carriers had significantly higher [11C]DASB binding in the caudate nucleus, but no significant differences in openness scores. Thus, high scores in personality facets indicative of cognitive flexibility and openness to change are associated with lower [11C]DASB binding. Lower abundance of 5-HTT sites may result in potentiation of serotonergic signaling, which occurs during treatment with SSRIs. We speculate that the set-point of serotonergic signaling in an individual represents a trade-off between flexibility and vulnerability when exposed to environmental stress. PMID- 19135153 TI - Cortical overlap of joint representations contributes to the loss of independent joint control following stroke. AB - The loss of independent joint control in the paretic upper limb is a cardinal sign of movement disorders following stroke. However, the underlying neural mechanisms for such a loss following stroke are still largely unknown. In order to investigate the possible contribution of altered sensorimotor cortical activity to the loss of independent joint control, we measured electroencephalographic (EEG) and torque signals during the generation of static shoulder/elbow torques. We found significant increases in the overlap of shoulder and elbow joint representations at the cortical level in stroke subjects as compared to control subjects. Linear regression results demonstrated significant associations between the cortical overlap of joint representations and the degree of the loss of independent joint control. Therefore, we conclude that an increased overlap of cortical representations for shoulder and elbow contributes to the expression of the loss of independent shoulder/elbow control of the paretic upper limb in chronic hemiparetic stroke survivors. PMID- 19135155 TI - Short-term adaptation to a simple motor task: a physiological process preserved in multiple sclerosis. AB - Short-term adaptation indicates the attenuation of the functional MRI (fMRI) response during repeated task execution. It is considered to be a physiological process, but it is unknown whether short-term adaptation changes significantly in patients with brain disorders, such as multiple sclerosis (MS). In order to investigate short-term adaptation during a repeated right-hand tapping task in both controls and in patients with MS, we analyzed the fMRI data collected in a large cohort of controls and MS patients who were recruited into a multi-centre European fMRI study. Four fMRI runs were acquired for each of the 55 controls and 56 MS patients at baseline and 33 controls and 26 MS patients at 1-year follow up. The externally cued (1 Hz) right hand tapping movement was limited to 3 cm amplitude by using at all sites (7 at baseline and 6 at follow-up) identically manufactured wooden frames. No significant differences in cerebral activation were found between sites. Furthermore, our results showed linear response adaptation (i.e. reduced activation) from run 1 to run 4 (over a 25 minute period) in the primary motor area (contralateral more than ipsilateral), in the supplementary motor area and in the primary sensory cortex, sensory-motor cortex and cerebellum, bilaterally. This linear activation decay was the same in both control and patient groups, did not change between baseline and 1-year follow-up and was not influenced by the modest disease progression observed over 1 year. These findings confirm that the short-term adaptation to a simple motor task is a physiological process which is preserved in MS. PMID- 19135156 TI - Prediction of visual field deficits by diffusion tensor imaging in temporal lobe epilepsy surgery. AB - Visual field deficits due to optic radiation injury are a common complication of temporal lobectomy in epilepsy surgery. In this prospective study, diffusion tensor imaging (DTI) based fiber tracking was performed on 48 patients who had temporal lobectomy for pharmaco-resistant epilepsy. Pre- and intra-operative DTI based fiber tracking was used to visualize the optic radiation and to predict the post-operative visual field defects. The course of the optic radiation could be successfully reconstructed by DTI based fiber tracking. There was a significant correlation between the fiber tracking estimation and the outcome of visual field deficits after surgery. The Receiver Operating Characteristic (ROC) curve analysis confirmed the accuracy and validity of prediction of the post-operative visual field deficits comparing pre- and intra-operative fiber tracking results. Intra-operative visualization of the optic radiation may help in avoiding post operative visual field deficits. PMID- 19135158 TI - Cancrincolidae (Copepoda, Harpacticoida) associated with land crabs: a semiterrestrial leaf of the ameirid tree. AB - Morphological evidence suggests harpacticoid copepods have recurrently entered into symbiosis with other crustaceans but only members of the family Cancrincolidae have successfully made the transition from marine habitats to semiterrestrial hosts. Cancrincolids are primarily amphi-Atlantic in distribution (with one outlier in the western Pacific) and typically inhabit the gill chambers of grapsoidean land crabs belonging to the families Grapsidae, Sesarmidae, Varunidae and Gecarcinidae. Morphologically, they are difficult to place because they exhibit unusual autapomorphies and the shared derived characters claimed to unite them with the primitively marine Ameiridae are equivocal. Both maximum parsimony and Bayesian inference solutions based on SSU rDNA gene sequences show topological congruence in placing the Cancrincolidae within the Ameiridae and in firmly resolving it as the sistergroup of taxa that have been reported as obligate or commensal associates of crayfish. This relationship is further supported by swimming leg sexual dimorphism and mandibular palp morphology. Morphological comparison with ameirid copepods revealed the majority of synapomorphies previously proposed in support of cancrincolid monophyly and familial distinctiveness can be attributed to heterochrony. The progressive evolution of cancrincolid associates appears to be largely concordant with the sequential adaptation to terrestriality by their grapsoidean hosts. The current amphi-Atlantic distribution of Cancrincola may suggest its possibly free-living ancestor had already assumed a virtually continuous distribution along the northern seaboard of Gondwana prior to the opening of the South Atlantic during the early Cretaceous, implying symbiotic relationships were established only much later when grapsoidean crabs started to emerge, radiate and diversify in the mid Tertiary (15-35 mya). The adoption of semi-terrestriality in cancrincolid copepods can be viewed as yet another independent attempt (incursion) to colonize low-salinity environments by members of the family Ameiridae. The possible sistergroup relationships of the Ameiridae and the position of the genus Argestigens Willey are discussed. PMID- 19135159 TI - Historical biogeography and speciation in the neotropical highlands: molecular phylogenetics of the jay genus Cyanolyca. AB - Phylogenetic relationships were studied in the genus Cyanolyca, an assemblage of jays distributed from Mexico south to Bolivia. Given its fragmented distribution along the humid forests of the Neotropics, the genus Cyanolyca is a model group for exploring hypotheses on biogeography and speciation. Phylogenetic analyses were based on two mitochondrial and three nuclear loci; taxon sampling includes all species in the genus and most subspecies. Maximum parsimony, maximum likelihood, and Bayesian analyses produced trees that were congruent and highly robust at both terminal and deep nodes of the phylogeny. Cyanolyca comprises two major clades: one contains the Mesoamerican "dwarf" jays, and the other consists of two main groups--C. cucullata+C. pulchra and the "core" South American species. Prior hypotheses of relationships were explored statistically using Maximum Likelihood and Bayesian approaches. Dispersal-Vicariance analysis revealed the importance of the Northern Andes as a major center for biological diversification, and the effects of dispersal across the Panamanian Land Bridge in the composition of South American and Mesoamerican avifaunas. Phylogenetic patterns are highly congruent with an allopatric mode of speciation. Implications of these results are discussed in the context of the biogeography of Neotropical montane forests. PMID- 19135157 TI - Differential visually-induced gamma-oscillations in human cerebral cortex. AB - Using intracranial electrocorticography, we determined how cortical gamma oscillations (50-150 Hz) were induced by different visual tasks in nine children with focal epilepsy. In all children, full-field stroboscopic flash-stimuli induced gamma-augmentation in the anterior-medial occipital cortex (starting on average at 31 ms after stimulus presentation) and subsequently in the lateral polar occipital cortex; minimal gamma-augmentation was noted in the inferior occipital-temporal cortex; occipital gamma-augmentation was followed by gamma attenuation in three children. Central-field picture-stimuli induced sustained gamma-augmentation in the lateral-polar occipital cortex (starting on average at 69 ms) and subsequently in the inferior occipital-temporal cortex in all children and in the posterior frontal cortex in three children; the anterior-medial occipital cortex showed no gamma-augmentation but rather gamma-attenuation. Electrical stimulation of the anterior-medial occipital cortex induced a phosphene in the peripheral-field or eye deviation to the contralateral side, whereas that of the lateral-polar occipital cortex induced a phosphene in the central-field. In summary, full-field, simple and short-lasting visual information might be preferentially processed by the anterior-medial occipital cortex, and subsequently by the lateral-polar occipital cortex. Gamma-attenuation following augmentation in the striate cortex might be associated with a relative refractory-period to flash-stimuli or feed-forward inhibition by other areas. Central-field complex visual information might be processed by a network involving the lateral-polar occipital cortex and the inferior occipital-temporal cortex. A plausible interpretation of posterior frontal gamma-augmentation during central-field picture stimuli includes activation of the frontal-eye-field for visual searching. Gamma-attenuation in the anterior-medial occipital cortex during central-field picture-stimuli might be associated with relative inattention to the peripheral visual field during central-field object visualization. PMID- 19135160 TI - Exploring the radiation of a diverse reef fish family: phylogenetics of the damselfishes (Pomacentridae), with new classifications based on molecular analyses of all genera. AB - The damselfishes (Perciformes, Pomacentridae) are an important family of marine reef fishes that occupy a range of ecological positions in the world's oceans. In order to determine the evolutionary pattern of their radiation, we used multiple methods to examine molecular data from 104 species representing all extant genera. The analysis of 4291 DNA nucleotides (1281bp were parsimony informative) from three nuclear genes (rag-1, rag-2 and bmp-4) and three mitochondrial genes (12s, 16s and nd3), produced well-resolved phylogenies with strong evidence for a monophyletic Pomacentridae, and support for five major damselfish clades. We found that the monotypic subfamily Lepidozyginae evolved early in the pomacentrid radiation. The placement of the genus Altrichthys indicates that brood care has evolved at least twice among the damselfishes. The subfamilies Chrominae and Pomacentrinae, and the genera Abudefduf, Chromis, Chrysiptera, Plectroglyphidodon, and Stegastes, were always found to be polyphyletic, and monophyly was rejected for the genus Amphiprion by almost every analysis. All phylogenetic studies of the Pomacentridae have indicated that their taxonomy is in need of revision at multiple levels. We provide a new classification scheme wherein each subfamily is now monophyletic, and this reorganization is consistent with all previous molecular studies of the damselfishes. The Chrominae are restricted to the genera Chromis and Dascyllus; the Pomacentrinae now represent a lineage of 16 genera that constitute a major, and relatively recent, radiation of coral reef fishes throughout the Indo-West Pacific; we erect the new subfamlies Abudefdufinae and Stegastinae; we relegate the anemonefishes (the Amphiprioninae sensu Allen) to the tribe Amphiprionini within the Pomacentrinae, and synonomize the genus Azurina with Chromis. PMID- 19135161 TI - RBP-Jkappa binds to and represses transcription of the p53 tumor suppressor gene. AB - The tightly regulated expression of p53 contributes to genomic stability and transcription of the p53 gene is induced prior to cells entering S-phase, possibly as a mechanism to insure a rapid p53 response in the event of DNA damage. We have previously described the cloning of an additional 1000bp of upstream p53 sequences that play a role in the regulated expression of p53, and identified that C/EBPbeta-2 participates in inducing p53 gene expression in a cell cycle regulated fashion. This report deals with the transcriptional regulator, RBP-Jkappa, an essential target of the Notch receptor signaling pathway. It binds to the p53 promoter in a cell cycle regulation fashion and also serves to repress p53 gene expression. We conclude from these findings that the coordinate expression of C/EBPbeta-2 and RBP-Jkappa may be linked to p53 transcription during G(0) and as cells move into S-phase. Because defects in the Notch signaling pathway have been implicated in carcinogenesis, aberrant RBP Jkappa expression and deregulated regulation of the p53 tumor suppressor could be an important step in some forms of cancers. PMID- 19135162 TI - The role of sulfur dioxide as an endogenous gaseous vasoactive factor in synergy with nitric oxide. AB - To explore the physiological role of endogenous gaseous sulfur dioxide (SO(2)) on vascular contractility and its underlying cellular and molecular mechanisms, vasodilation experiment of isolated rat thoracic aortic rings by gaseous SO(2) was carried out and the signal transduction pathways involved in the vascular effects of SO(2) were investigated. In the present study, SO(2) gas and SO(2) gas bubbled solution (SO(2) stock solution) were first used to relax vascular tissues. The results show: (1) Gaseous SO(2) relaxed rat thoracic aortic rings in a dose-dependent manner (from 1 to 2000microM). The vasorelaxant effect of SO(2) at physiological relevant and low concentrations (<450microM) was endothelium dependent, and at high concentrations (>500microM) was endothelium-independent. (2) The vasorelaxation by addition of SO(2) stock solution (final concentrations 2mM) was actually caused by SO(2) molecules, not by sulfite or bisulfite, and the characteristic of vasorelaxation by SO(2) was different from that of sulfite and bisulfite. (3) The vasorelaxant effect of SO(2) was not due to the altered neurotransmitter release from the autonomous or nonadrenergic and noncholinergic (NANC) nerve endings, also not due to superoxide and hydrogen peroxide produced in the vascular tissues, also disapproving the involvement of prostaglandin, PKC, beta-adrenoceptor and cAMP pathways. (4) The vasorelaxant effect of SO(2) at the physiological relevant and low concentrations was mediated by the cGMP pathway. (5) There was the synergistic effect on smooth muscle relaxation between much lower concentrations of SO(2) (3microM) and NO (3 or 5nM). These findings led to the conclusions: endogenous gaseous SO(2) was a vasoactive factor, which might regulate vascular smooth muscle tone in synergy with NO. PMID- 19135163 TI - Tissue and matrix influences on airway smooth muscle function. AB - Asthma is characterized by structural changes in the airways - airway remodelling. These changes include an increase in the bulk of the airway smooth muscle (ASM) and alterations in the profile of extracellular matrix (ECM) proteins in the airway wall. The mechanisms leading to airway remodelling are not well understood. ASM cells have the potential to play a key role in these processes through the production and release of ECM proteins. The ASM cells and ECM proteins are each able to influence the behaviour and characteristics of the other. The modified ECM profile in the asthmatic airway may contribute to the altered behaviour of the ASM cells, such responses to ECM proteins are modulated through the cell surface expression of integrin receptors. ASM cells from asthmatic individuals express different levels of some integrin subunits compared to nonasthmatic ASM cells, which have the potential to further influence their responses to the ECM proteins in the airways. ECM homeostasis requires the presence and activation of matrix metalloproteinases and their tissue inhibitors, which in turn modulate the interaction of the ASM cells and the ECM proteins. Furthermore, the complex interactions of the ASM cells and the ECM in the asthmatic airways and the role played by external stimuli, such as viral infections, to modulate airway remodelling are currently unknown. This review summarises our current understanding of the influence of the ECM on ASM function. PMID- 19135164 TI - Conserved role of cyclic nucleotides in the regulation of ecdysteroidogenesis by the crustacean molting gland. AB - Molting processes in crustaceans are regulated by ecdysteroids produced in the molting gland (Y-organ), and molting is indirectly controlled by circulating factors that inhibit the production of these polyhydroxylated steroids. Two of these regulatory factors are the neuropeptides molt-inhibiting hormone (MIH) and crustacean hyperglycemic hormone (CHH). CHH appears to inhibit ecdysteroidogenesis in the Y-organ through the activation of a receptor guanylyl cyclase. The signaling pathway activated by MIH, however, remains a subject of controversy. It is clear that neuropeptides inhibit ecdysteroidogenesis by simultaneously suppressing ecdysteroid biosynthetic processes, protein synthesis, and uptake of high density lipoproteins. Data demonstrate that cAMP is the primary regulator of critical catabolic, anabolic, and transport processes, which ultimately support the capacity for ecdysteroid production by the Y-organ. While cAMP also regulates acute ecdysteroidogenesis to some extent, data indicate that cGMP is the primary signaling molecule responsible for acute inhibition by neuropeptides. It is clear that the regulatory roles filled by cAMP and cGMP are conserved among decapod crustaceans. It is unknown if these complementary second messengers are linked in a single signaling pathway or are components of independent pathways activated by different factors present in extracts of eyestalk ganglia. PMID- 19135165 TI - Feeding restriction alters expression of some ATP related genes more sensitively than the RNA/DNA ratio in zebrafish, Danio rerio. AB - The growth rate of fish shows extensive plasticity in response to various environments. Metabolic responses of fish to excessive nutritional shortages such as starvation have been reported, but the effects of moderate nutrient shortage remain unclear. We examined expression levels of some genes related to ATP metabolism and to myogenesis, the RNA/DNA ratio, and the protein/DNA ratio of fish under different feeding conditions: a diet of 212-432% (frequent feeding, FR) or 32-82% (restricted feeding, RE) of initial body weight per week was supplied. The expression levels of nucleoside diphosphate kinase (NDK)-Z2, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and myogenin genes of RE fish were higher than those of FR fish, although the RNA/DNA ratio and the protein/DNA ratio were unaffected by the feeding amount. Moreover, expression levels of NDK Z2 and GAPDH were upregulated to a greater extent than those for myogenin and myostatin 1 under restricted feeding. Together, our results show that gene expression is more sensitive to nutrient conditions of fish than traditional indicators such as the RNA/DNA ratio. The ATP metabolic system is more sensitive to moderate nutrient shortages than the myogenic system. PMID- 19135166 TI - Molecular characterization, gene expression analysis and biochemical properties of alpha-amylase from the disk abalone, Haliotis discus discus. AB - The present study reports the molecular characterization, cloning, expression, and biochemical characterization of alpha-amylase identified from the disk abalone, Haliotis discus discus cDNA library. The full length of the alpha amylase cDNA was 1650 bp, and it encoded a polypeptide of 511 amino acids. The predicted HdAmyI molecular mass of mature protein was 54 kDa and the estimated isoelectric point (pI) was 8.3. The alpha-amylase gene showed its characteristic motifs, catalytic sites, substrate binding sites and conserved regions with other known species of alpha-amylases. Purified recombinant HdAmyI exhibited a relatively low activity of 0.1 U/mg protein towards 1% starch. HdAmyI had an optimum temperature and pH of 50 degrees C and 6.5, respectively. It also demonstrated stability in a wide range of temperatures and pH. Tissue-specific mRNA expression results showed that HdAmyI is expressed only in the digestive tract and hepatopancreas, with the highest levels in the hepatopancreas. Over 8 weeks of starvation, alpha-amylase transcription was decreased significantly relative to basal levels. However, after starvation, mRNA transcription was increased and returned to normal level by the 2nd week of feeding, suggesting that the alpha-amylase mRNA expression changes according to variations in food availability at the transcriptional level in disk abalone. PMID- 19135167 TI - MUC1 mucin interacts with calcium-modulating cyclophilin ligand. AB - MUC1 is an integral membrane glycoprotein expressed on epithelial and hematopoietic cells with a COOH-terminus (CT) that mediates intracellular signal transduction. To better understand MUC1-dependent signaling, we searched for proteins binding to its CT using the yeast two-hybrid system with the MUC1 CT as bait and a human epithelial cell cDNA library as prey. Of the six positive clones identified, all encoded calcium-modulating cyclophilin ligand (CAML). The MUC1 CT interacted with CAML in transformed yeast cells as revealed by growth on selective media and in situ X-alpha-galactosidase activity. Binding of the MUC1 CT to CAML in human epithelial cells was confirmed by reciprocal coimmunoprecipitations, confocal microscopy, protein crosslinking, and coupled transcription/translation analyses. By deletion mutagenesis, the NH(2)-terminus of CAML was responsible for binding to the MUC1 CT. Finally, transfection of cells with plasmids encoding MUC1 and CAML increased intracellular calcium levels compared with cells transfected with either plasmid alone, suggesting a possible biological significance of the MUC1-CAML interaction. PMID- 19135168 TI - Monitoring the amyloid beta-peptide in vivo--caveat emptor. AB - As a wave of 'disease modifying' (DM) therapies for Alzheimer's disease (AD) progresses towards the later stages of clinical development, an evaluation of our ability to measure relevant pharmacodynamic effects of such therapies is warranted. Reducing accumulation of amyloid beta (Abeta)-peptide in the brain parenchyma is the primary objective of most current DM approaches. Although a number of methods are available to measure Abeta in blood, cerebrospinal fluid (CSF) and the cerebrum, putative DM-induced changes in the levels of the peptides may not be fully captured, and the reasons for any such changes are not fully understood. Additional candidate biofluid (tau and isoprostanes) and imaging (MRI, FDG-PET) measures may provide alternative supporting evidence of drug activity and subsequent clinical efficacy in patient populations. PMID- 19135169 TI - The genetic toxicology of methylphenidate hydrochloride in non-human primates. AB - The studies presented in this work were designed to evaluate the genetic toxicity of methylphenidate hydrochloride (MPH) in non-human primates (NHP) using a long term, chronic dosing regimen. Thus, approximately two-year old, male rhesus monkeys of Indian origin were orally exposed to MPH diluted in the electrolyte replenisher, Prang, five days per week over a 20-month period. There were 10 animals per dose group and the doses were (1) control, Prang only, (2) low, 0.15 mg/kg of MPH twice per day increased to 2.5mg/kg twice per day and (3) high, 1.5 mg/kg of MPH twice per day increased to 12.5 mg/kg twice per day. Blood samples were obtained from each animal to determine the base-line serum levels of MPH and the major metabolite of MPH in NHP, ritalinic acid (RA). In addition, the base line frequency of micronucleated erythrocytes (MN-RETs) by flow cytometry, HPRT mutants by a lymphocyte cloning assay, and chromosome aberrations by FISH painting were determined from peripheral blood samples. Once dosing began, the serum levels of MPH and its major metabolite, RA, were determined monthly. The MN RET frequency and health parameters (CBC, serum chemistries) were also determined monthly. HPRT mutant and chromosome aberration frequencies were measured every three months. CBC values and serum chemistries, with the exception of alanine amino transferase, were within normal limits over the course of drug exposure. The final plasma levels of MPH were similar to those produced by the pediatric dose of 0.3 microg/ml. No significant increases in the frequencies of MN-RETs, HPRT mutants, or chromosome aberrations were detected in the treated animals compared to the control animals over the 20-month exposure period. PMID- 19135170 TI - Establishment of an in vitro transgene expression system in epimastigotes of Trypanosoma congolense. AB - Trypanosoma congolense epimastigote forms (EMFs) adhere to the tsetse fly proboscis, proliferate, and differentiate into animal-infective metacyclic forms (MCFs). This differentiation step, called metacyclogenesis, is indispensable for the cyclical transmission of the parasite. Although an in vitro metacyclogenesis culture system was established several decades ago, few genetic tools have been utilized to investigate the molecular mechanisms underlying T. congolense metacyclogenesis. This study established a transgene expression system using an in vitro derived EMF of T. congolense IL3000, and the transgenic EMF successfully underwent metacyclogenesis in vitro. The newly constructed expression vector pSAK was designed for integration into the alpha-beta tubulin locus, which is tandemly arranged in the T. congolense genome. The expression cassette of pSAK/enhanced green fluorescent protein (eGFP) was transfected into the EMF by electroporation. An EMF expressing eGFP was successfully generated and differentiated into an MCF that constitutively expressed eGFP. The in vitro metacyclogenesis system in combination with the transgenic EMF technique will be important tools to investigate the molecular mechanisms of metacyclogenesis. PMID- 19135173 TI - Cognitive and learning sciences in biomedical and health instructional design: A review with lessons for biomedical informatics education. AB - Theoretical and methodological advances in the cognitive and learning sciences can greatly inform curriculum and instruction in biomedicine and also educational programs in biomedical informatics. It does so by addressing issues such as the processes related to comprehension of medical information, clinical problem solving and decision-making, and the role of technology. This paper reviews these theories and methods from the cognitive and learning sciences and their role in addressing current and future needs in designing curricula, largely using illustrative examples drawn from medical education. The lessons of this past work are also applicable, however, to biomedical and health professional curricula in general, and to biomedical informatics training, in particular. We summarize empirical studies conducted over two decades on the role of memory, knowledge organization and reasoning as well as studies of problem-solving and decision making in medical areas that inform curricular design. The results of this research contribute to the design of more informed curricula based on empirical findings about how people learn and think, and more specifically, how expertise is developed. Similarly, the study of practice can also help to shape theories of human performance, technology-based learning, and scientific and professional collaboration that extend beyond the domain of medicine. Just as biomedical science has revolutionized health care practice, research in the cognitive and learning sciences provides a scientific foundation for education in biomedicine, the health professions, and biomedical informatics. PMID- 19135172 TI - Rainbow trout (Oncorhynchus mykiss) and ultra-low dose cancer studies. AB - Cancer risk assessment utilizing rodents requires extrapolation across five orders of magnitude to estimate the Virtually Safe Dose (VSD). Regulatory agencies rely upon the Linear Extrapolated Dose (LED) except when sufficient information on mechanism of action justifies alternative models. Rainbow trout (Oncorhynchus mykiss) has been utilized at Oregon State University as a model for human cancer for forty years. Low cost and high capacity, made possible by our unique facility, along with low spontaneous background and high sensitivity, allow design and conduct of statistically challenging studies not possible in rodents. Utilization of custom microarrays demonstrates similarities in gene expression in trout and human hepatocellular carcinoma (HCC). We have completed one study employing over 42,000 trout with dibenzo[a,l]pyrene (DBP) and determined the dose resulting in 1 additional cancer in 5000 animals, a 50-fold enhancement over the mouse ED(01) study. Liver tumor incidence at low dose deviated significantly from linearity (concave down), whereas, DBP-DNA adductions deviated slightly (convex up). A second study is underway with aflatoxin B(1) (AFB(1)). Results to date indicate AFB(1) at low dose, in contrast to DBP, elicits a linear dose-response function on the log-log scale which falls below the LED with a slope slightly greater than 1.0. Such studies demonstrate the statistical power of the trout cancer model and strengthen the case for incorporation of these data-sets into risk assessment for these environmental human carcinogens. PMID- 19135175 TI - Occult hepatocellular carcinoma metastatic to the mandible. PMID- 19135174 TI - Role of angiotensin II in the response to endothelin-1 of goat cerebral arteries after ischemia-reperfusion. AB - As angiotensin II may underlie the deleterious effects of some vascular diseases, we have examined the role of this peptide on the cerbrovascular endothelin-1 action after ischemia-reperfusion. In anesthetized goats, 1 hour-occlusion followed by 1 hour-reperfusion of the left middle cerebral artery (MCA) was induced, and then segments 3-mm in length from branches of the right MCA (control) and the left MCA (ischemic) were obtained for isometric tension recording. Endothelin-1 (10(-11)-10(-7) M) produced a contraction that was higher in ischemic than in control arteries, and in control but not in ischemic arteries this contraction was potentiated by angiotensin II (10(-7) M). Losartan (3 x 10( 6) M), antagonist of AT1 receptors, did not affect the response to endothelin-1 in control arteries, but reduced it both in ischemic arteries and angiotensin II treated control arteries. PD123,319 (3 x 10(-6) M), antagonist of AT2 receptors, or the inhibitor of nitric oxide synthesis L-NAME (10(-4) M) did not alter the arterial effects of endothelin-1. Therefore, angiotensin II may potentiate the constriction to endothelin-1 in normal cerebral arteries by activating AT1 receptors. The observed cerebrovascular increased response to endothelin-1 after ischemia-reperfusion might be related in part to activation of AT1 receptors under this condition. PMID- 19135177 TI - Morphological evidence suggests homoploid hybridization as a possible mode of speciation in the Triatominae (Hemiptera, Heteroptera, Reduviidae). AB - All known significant insect vectors of Trypanosoma cruzi are members of the Reduviidae, subfamily Triatominae. Infections with this parasite are the cause of Chagas disease, the single most costly parasitic disease in the Western Hemisphere. The Triatominae are almost completely restricted to the Americas, with >130 species distributed in several foci of species richness and endemism; nevertheless, the processes involved in the diversification of this group remain poorly understood. The Triatoma brasiliensis species complex was recently proposed based on geography, morphology, ecology, and molecular data, and is believed to comprise two species and two subspecies. Here, we report results from a broad series of studies, in which first-generation offspring of experimental crosses were studied in terms of wing morphometry and phylogenetic position. Morphometrics, morphological, ecological and geographic analyses were consistent with the hypothesis of T. brasiliensis macromelasoma as a product of hybridization between two others (T. brasiliensis brasiliensis and T. juazeirensis). Although evidence is supportive of the hypothesis of speciation via hybridization as a mode of triatomine diversification, the case is not as-yet conclusive, and confirmation via molecular markers is necessary. PMID- 19135178 TI - Suppression of inducible nitric oxide synthase by (-)-isoeleutherin from the bulbs of Eleutherine americana through the regulation of NF-kappaB activity. AB - Eleutherine americana Merr. et Heyne (Iridaceae) has been used as a folk medicine for the treatment of coronary disorders and wound-healing. In our previous phytochemical study, several pyranonaphthoquinoids, including (-)-isoeleutherin, were isolated from the bulbs of E. americana. Because inhibitors of inducible nitric oxide synthase (iNOS) might be useful as anti-inflammatory agents, we investigated the potential of pyranonaphthoquinoids to inhibit iNOS activity. We found that (-)-isoeleutherin inhibits lipopolysaccharide (LPS)-stimulated induction of nitric oxide (NO) in a dose-dependent manner (IC(50)=7.4 microM). Using western blots and reverse transcription-polymerase chain reaction, we showed that (-)-isoeleutherin suppresses the expression of iNOS protein and mRNA. In addition, (-)-isoeleutherin inhibited the expression of various cytokines such as interleukin-1beta and interferon-beta. Activation of the transcriptional activity of NF-kappaB by LPS was also inhibited by treatment with (-) isoeleutherin, suggesting that (-)-isoeleutherin-mediated suppression of iNOS expression is associated with the regulation of transcription factor NF-kappaB. These findings suggest that (-)-isoeleutherin might be a novel anti-inflammatory agent, and that it may work by inhibiting NFkB activation in macrophages. PMID- 19135179 TI - Requirement of L-selectin for gammadelta T lymphocyte activation and migration during allergic pleurisy: co-relation with eosinophil accumulation. AB - Intra-thoracic antigenic challenge (ovalbumin, 12.5 microg/cavity) led to increased numbers of gammadelta T lymphocytes in pleural cavities, blood and thoracic lymph nodes in sensitized mice within 48 h. Part of these cells expressed CD62L, which increased on gammadelta T cell surfaces obtained from lymph nodes after ovalbumin (OVA) challenge. Selectin blockade by fucoidan pre treatment (10 mg/kg, i.v.) impaired in vivo increase in CD25(+) and c-fos(+) gammadelta T cell numbers in lymph nodes, indicating a role for selectins on gammadelta T lymphocyte activation and proliferation. In vivo selectin blockade by fucoidan or alpha-CD62L mAb (200 microg/mice, i.p.) also inhibited OVA-induced gammadelta T cell accumulation in pleural cavities. Confirming the direct effect of CD62L on gammadelta T cell transmigration, the migration of i.v. adoptively transferred CFSE-labeled gammadelta T lymphocytes into pleural cavities of challenged recipient mice was impaired by fucoidan ex vivo treatment. It is noteworthy that eosinophil influx was also impaired in those mice, indicating that reduced eosinophil migration by CD62L in vivo blockade depended on gammadelta T cell migration via CD62L molecules. Accordingly, pleural gammadelta T lymphocytes from fucoidan-treated mice presented reduced OVA-induced IL-5 and CCL11 production. Supporting these data, the depletion of Vgamma4 T lymphocytes, which are pulmonary gammadelta T cells, decreased OVA-induced eosinophil influx into allergic site. Such results demonstrate that CD62L is crucial for the activation of gammadelta T cells in lymph nodes, for their migration into inflamed tissue and for the modulation of eosinophil influx during allergic response. PMID- 19135180 TI - Benefits and burden of the maternally-mediated immunological imprinting. AB - The ontogenetic development of both the immune and the nervous system entirely depend on external environmental signals that induce a lifelong learning process. The resulting collective immunological knowledge about the external world is transmitted in an epi-genetic fashion to the offspring, but only from the maternal and not the paternal side, with maternal IgG as the main transgenerational vector. As products of thymus-dependent responses, maternal IgG have undergone immune maturation by somatic hypermutations and are, therefore, acquired immunological phenotypes representing a great deal of the mother's immunological experience. During a limited neonatal imprinting period, maternal antibodies induce T cell-dependent idiotypic responses. These exert up to life long determinative influences which may even be dominant over seemingly genetic predispositions. Such long-term immunological imprinting effects can be detected as (a) selection of the adult T and B cell repertoires, (b) anti-microbial protection by antigen-reactive antibodies (idiotypes) and anti-idiotypes, (c) allergen-specific suppression of IgE responsiveness by allergen-reactive IgG idiotype or corresponding anti-idiotype and (d) induction of autoimmune diseases by maternally-derived autoantibodies. Hence, immunological imprinting by maternal IgG antibodies will mostly be beneficial, but in case of autoantibodies can also be a burden for the initial development of the nascent immune system. PMID- 19135181 TI - Different strains of mice present distinct lung tissue mechanics and extracellular matrix composition in a model of chronic allergic asthma. AB - The impact of genetic factors on asthma is well recognized but poorly understood. We tested the hypothesis that different mouse strains present different lung tissue strip mechanics in a model of chronic allergic asthma and that these mechanical differences may be potentially related to changes of extracellular matrix composition and/or contractile elements in lung parenchyma. Oscillatory mechanics were analysed before and after acetylcholine (ACh) in C57BL/10, BALB/c, and A/J mice, subjected or not to ovalbumin sensitization and challenge. In controls, tissue elastance (E) and resistance (R), collagen and elastic fibres' content, and alpha-actin were higher in A/J compared to BALB/c mice, which, in turn, were more elevated than in C57BL/10. A similar response pattern was observed in ovalbumin-challenged animals irrespective of mouse strain. E and R augmented more in ovalbumin-challenged A/J [E: 22%, R: 18%] than C57BL/10 mice [E: 9.4%, R: 11%] after ACh In conclusion, lung parenchyma remodelled differently yielding distinct in vitro mechanics according to mouse strain. PMID- 19135182 TI - Stimulation of the rat medullary raphe nuclei induces differential responses in respiratory muscle activity. AB - Neural control circuits that coordinate the motor activity of the diaphragm (DIA) and the geniohyoid muscle (GH) are potentially involved in pathological conditions such as various forms of sleep apnea. Here we investigated a differential role of the raphe magnus (RMg), pallidus (RPa) and the obscurus (ROb) nuclei in the neural control of DIA and GH muscle activity in rats under volatile anesthesia. In order to characterize a topographical organization of the raphe nuclei we analyzed changes in DIA and GH during high-frequency stimulation (HFS, 10-130 Hz, 60 micros pulse width, 40-160 microA, 30s). HFS of the RMg and the ROb induced apnea, in the latter case apnea was associated with massive tonic discharge in the GH. By contrast, HFS of the RPa induced tachypnea. At caudal stimulation sites the tachypnea was accompanied by tonic DIA activity and cessation of GH. These data suggest a differential distribution of inhibitory and excitatory drives of DIA and GH muscles within distinct raphe nuclei. PMID- 19135183 TI - Thioredoxin targets in plants: the first 30 years. AB - The turn of the century welcomed major developments in redox biology. In plants, proteomics made possible the identification of proteins linked to thioredoxin (Trx), initially in chloroplasts and then other cell compartments. Two procedures, one based on thiol specific probes and the other on mutant Trx proteins, facilitated the labeling or isolation of potential Trx targets that were later identified with proteomic approaches. As a result, the number of targets in land plants increased 10-fold from fewer than 40 to more than 400. Additional targets have been identified in green algae and cyanobacteria, making a grand total of 500 in oxygenic photosynthetic organisms. Collectively these proteins have the potential to influence virtually every major process of the cell. A number of laboratories currently seek to confirm newly identified Trx targets by biochemical and genetic approaches. Almost certainly many new targets become redox active during oxidative stress, enabling the plant to cope with changing environments. Under these conditions, certain targets may be glutathionylated or nitrosylated such that reversion to the original reduced state is facilitated not only by Trx, but also, in some cases preferably, by glutaredoxin. When judging changes linked to Trx, it is prudent to recognize that effects transcend classical light/dark or oxidative regulation and fall in other arenas, in some cases yet to be defined. While future work will continue to give insight into functional details, it is clear that Trx plays a fundamental role in regulating diverse processes of the living cell. PMID- 19135184 TI - Trabecular bone strength predictions using finite element analysis of micro-scale images at limited spatial resolution. AB - Advances in micro-scanning technology have led to renewed clinical interest in the ability to predict bone strength using finite element (FE) analysis based on images with resolutions in the range of 80 microm. Using such images, we sought to determine whether predictions of yield stress provided by nonlinear FE models could improve correlations with bone strength as compared to the use of predictions of elastic modulus from linear FE models, and if this effect depended on voxel size or bone volume fraction. Linear and nonlinear FE analyses were conducted for 46 trabecular cores from three human anatomic sites using element sizes ranging from 20 to 120 microm to obtain measures of apparent yield stress and elastic modulus, and these measures were correlated against the predicted yield stress from the 20 microm models (assumed to be the gold standard strength for this study). Results indicated that when considering all samples and any resolution, yield stress and elastic modulus were both excellent predictors of strength (R2>0.99). When only low-density samples (BV/TV<0.15) were considered, yield stress was better correlated with 20 microm-strength than was elastic modulus (R2 increased from 0.93 to 0.99 at 40 microm and from 0.90 to 0.95 at 80 microm). However, at a voxel size of 120 microm, the predictive ability of yield stress was slightly less than that of stiffness, likely due to the large convergence-related errors that could develop with larger element sizes. As expected, a limit was observed in the ability of elastic modulus to predict strength--the predictive ability of elastic modulus measured at 20 microm was comparable to that of yield strength at 80 microm. We also found that strength predictions from FE models at clinical-type resolutions had nearly the same power to detect bone quality effects via variations in strength-density relationships as did high-resolution models. We conclude that nonlinear FE models can account for additional variations in strength relative to linear models when using images at resolutions of approximately 80 microm and less, and such models offer a promising method for examining microarchitecture-related bone quality effects associated with aging, disease, and treatment. PMID- 19135185 TI - Effects of differences in office chair controls, seat and backrest angle design in relation to tasks. AB - In this study the influence of chair characteristics on comfort, discomfort, adjustment time and seat interface pressure is investigated during VDU and non VDU tasks: The two investigated office chairs, both designed according to European and Dutch standards are different regarding: 1) seat cushioning and shape, 2) backrest angle and 3) controls. Thirty subjects in total, both male and female, participated in two experiments: twenty in the first and ten in the second. Significant differences are found for ease of adjustment and adjustment time of controls, independent of the tasks. Related to tasks, a significant difference was found for the backrest range of motion. For non-VDU tasks a larger range of backrest motion was preferred by 70% of the subjects. The chair design differences were most clear for comfort and adjustment time of controls, followed by comfort of backrest angle. No differences are found between seat pan comfort and discomfort, first impressions and peak interface pressure. PMID- 19135186 TI - Influence of the location of the parotid duct orifice on oral clearance. AB - OBJECTIVES: To determine the location of the parotid duct orifice in relation to the maxillary molars and its influence on oral clearance on the buccal surfaces of the maxillary molars. METHODS: A 2-mm hole was made at the centre of an adhesive therapeutic agent for aphthous stomatitis and the agent was placed on the mucosa so that the hole matched the parotid duct orifice. To locate the orifice, an impression of the buccal tooth surfaces and mucosa around the agent was taken with the teeth in centric occlusion. To evaluate the oral clearance rate, 12 subjects who displayed the parotid duct orifice within 1S.D. of the mean values obtained from the original 35 subjects were selected. 1% agar containing 1 mol/l potassium chloride was placed into three cylinders positioned horizontally, 6mm apart, in an acrylic holder centred over the mean duct location. The diffusion chambers were taken from the mouth at selected time intervals and the gel transferred quantitatively to flasks containing 300 ml of 100 ppm NaCl, which was assayed for potassium by atomic absorption spectrophotometry. Half-times for clearance were calculated. RESULTS: The mean location of the parotid duct orifice was -0.4 mm (range -7.5 to +6.1 mm) mesial to the contact surface between the maxillary first and second molars (where negative values indicate mesial and positive values distal) and 7.2 mm (range +3.8 to +10.4 mm) above a line touching the buccal cusps of the upper molars. The clearance half-time values were shortest for the central cylinder whether salivary flow was unstimulated or stimulated and when flow was unstimulated the clearance half-time was shorter for the mesial than the distal cylinder. CONCLUSION: The degree of individual variation in the location of the parotid duct orifice is great and its exact location will markedly affect oral clearance at different positions on the buccal surfaces of the upper molars. PMID- 19135187 TI - Implicit self-esteem in bulimia nervosa. AB - Implicit and explicit self-esteem were compared in a group of female participants with bulimia nervosa or binge eating disorder (n=20) and a healthy control group (n=20). Lower explicit and a less positive implicit self-esteem bias in the clinical group was predicted. Participants completed a self-esteem implicit association test and two explicit self-esteem measures. The eating disordered group had lower explicit self-esteem, but a more positive implicit self-esteem bias than controls. The results are discussed in relation to the idea that discrepancies between implicit and explicit self-esteem reflect fragile self esteem and are related to high levels of perfectionism, which is associated with eating disorders. PMID- 19135188 TI - Evidence of a biological effect of light therapy on the retina of patients with seasonal affective disorder. AB - BACKGROUND: Retinal sensitivity anomalies have been reported in patients affected by seasonal affective disorder (SAD). We used the electroretinogram (ERG) to assess seasonal change in retinal function in patients with SAD and healthy participants, as well as in patients following 4 weeks of light therapy. METHODS: ERG assessments were obtained in 22 SAD patients (2 men, 20 women, mean age 31 +/ 9 years) in the fall/winter season before and after 2 and 4 weeks of light therapy and in summertime. Matched healthy participants (2 men, 14 women; mean age 29 +/- 8 years) were evaluated once in the fall/winter and once in summer. The 29-item Structured Interview Guide for the Hamilton Depression Rating Scale, Seasonal Affective Disorder version was administered. Standard ERG parameters were derived from the photopic and scotopic luminance response functions. Salivary melatonin concentration during ERG was assessed in both groups but during fall/winter assessments only. RESULTS: A significantly lower cone ERG maximal amplitude and lower rod sensitivity was found in SAD patients before light therapy compared with healthy participants. Following 4 weeks of light therapy, a normalization of cone and rod ERG function occurred. ERG parameters in the summer and melatonin concentrations in fall/winter were not significantly different between groups. CONCLUSIONS: Depressed patients with SAD demonstrate ERG changes in the winter compared with healthy comparison subjects with lower rod retinal sensitivity and lower cone maximal amplitude. These changes normalized following 4 weeks of light therapy and during the summer, suggesting that ERG changes are state markers for SAD. PMID- 19135189 TI - Fate of THMs and HAAs in low TOC surface water. AB - A total of 30 conventional surface water treatment plants (WTPs) implementing prechlorination and postchlorination simultaneously from different regions in Korea were investigated to assess formation and removal of THMs and HAA(5). All water was low in total organic carbon (TOC) ranging from 0.74 to 6.20 mg/L with an average of 1.63 mg/L. The ranges of THMs and HAA(5) levels were 4.5-84.3 microg/L and 1.5-90.8 microg/L, respectively. THMs concentration was more sensitive to water temperature than HAA(5) and the ratio of THMs in summer over winter was 2.06. The sum of dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) was 97% of HAA(5). The extent of formation and speciation of DBPs varied greatly by season and geography. The concentration of DCAA and TCAA of the finished water was comparable on a yearly base, but more TCAA was noticed in summer and the opposite trend was noticed in winter. This can be caused by different biodegradability in the sand filter between DCAA and TCAA that formed through prechlorination. Investigation on the removal of preformed DBPs in the GAC filter-adsorber (FA) revealed that breakthrough of THMs and HAA(5) was noticed after 3 months of operation. However, gradual improvement (>90%) in HAA(5) removal was observed again after breakthrough, which could be attributable to biodegradation. Heterotrophic plate counts confirmed active biological activity in the GAC FA. PMID- 19135190 TI - Maternal exposure to ambient air pollutants and risk of congenital anomalies. AB - Studies have suggested an association between maternal exposure to ambient air pollution and risk of congenital anomaly. The aim of this study is to investigate the association between exposure to black smoke (BS; particulate matter with aerodynamic diameter <4 microg/m(3)) and sulphur dioxide (SO(2)) during the first trimester of pregnancy and risk of congenital anomalies. We used a case-control study design among deliveries to mothers resident in the UK Northern health region during 1985-1990. Case data were ascertained from the population-based Northern Congenital Abnormality Survey and control data from national data on all births. Data on BS and SO(2) from ambient air monitoring stations were used to average the total pollutant exposure during the first trimester of pregnancy over the daily readings from all monitors within 10 km of the mother's residence. Logistic regression models estimated the association via odds ratios. A significant but weak positive association was found between nervous system anomalies and BS (OR=1.10 per increase of 1000 microg/m(3) total BS; 95% CI: 1.03, 1.18), but not with other anomaly subtypes. For SO(2), a significant negative association was found with congenital heart disease combined and patent ductus arteriosus: OR significantly <1 for all quartiles relative to the first quartile. The relationship between SO(2) levels and other anomaly subtypes was less clear cut: there were either no significant associations or a suggestion of a U-shaped relationship (OR significantly <1 for moderate compared to lowest levels, but not with high SO(2) levels). Overall, maternal exposure to BS and SO(2) in the Northern region had limited impact on congenital anomaly risk. Studies with detailed exposure assessment are needed to further investigate this relationship. PMID- 19135191 TI - Successful outcome after preimplantation genetic screening and very delayed embryo transfer. AB - We report the positive outcome of a delayed single ET of a preimplantation genetic screening-defined embryo in an otherwise routine case of IVF-ET. To our knowledge, this is the first report of a fresh ET 48 hours longer than what is generally considered the limit of days after egg retrieval (six) to safely perform an ET. PMID- 19135192 TI - Immunization with Bin1b decreases sperm motility with compromised fertility in rats. AB - OBJECTIVE: To evaluate the contraceptive ability of a synthetic Bin1b peptide in vivo in the rat. DESIGN: Basic research. SETTING: University laboratory animal service center. ANIMAL(S): A peptide-based immunization model was developed; rats were injected with the Bin1b specific peptide. INTERVENTION(S): A synthetic peptide segment, MCRSGERKGDICSDP-conjugated with KLH (Bin1b), was used to immunize male wistar rats. Freund's complete adjuvant was used as a control. MAIN OUTCOME MEASURE(S): Anti-Bin1b levels in sera were evaluated by enzyme-linked immunosorbent assay (ELISA). Anti-Bin1b and control antisera were used to evaluate sperm function inhibition in vitro. The fertility of immunized rats was determined by mating experiment. The testis and epididymides were analyzed by histology. RESULT(S): Histological studies showed no evidence of orchitis or epididymitis in Bin1b-immunized animals. ELISA results revealed that the titers of anti-Bin1b antibodies in serum increased with the immunization process in immunized rats. Sperm recovered from the corpus epididymidis of the Bin1b immunized animals exhibited a significant decrease in motility. Immunization of Bin1b also caused a reduction (25%) in fertility after the mating experiment. CONCLUSION(S): The present study has demonstrated that immunization with Bin1b peptide specifically interferes with sperm motility, resulting in a compromised fertilizing capacity of sperm. PMID- 19135193 TI - Exsanguination in trauma: A review of diagnostics and treatment options. AB - Trauma patients with haemorrhagic shock who only transiently respond or do not respond to fluid therapy and/or the administration of blood products have exsanguinating injuries. Recognising shock due to (exsanguinating) haemorrhage in trauma is about constructing a synthesis of trauma mechanism, injuries, vital signs and the therapeutic response of the patient. The aim of prehospital care of bleeding trauma patients is to deliver the patient to a facility for definitive care within the shortest amount of time by rapid transport and minimise therapy to what is necessary to maintain adequate vital signs. Rapid decisions have to be made using regional trauma triage protocols that have incorporated patient condition, transport times and the level of care than can be performed by the prehospital care providers and the receiving hospitals. The treatment of bleeding patients is aimed at two major goals: stopping the bleeding and restoration of the blood volume. Fluid resuscitation should allow for preservation of vital functions without increasing the risk for further (re)bleeding. To prevent further deterioration and subsequent exsanguinations 'permissive hypotension' may be the goal to achieve. Within the hospital, a sound trauma team activation system, including the logistic procedure as well as activation criteria, is essential for a fast and adequate response. After determination of haemorrhagic shock, all efforts have to be directed to stop the bleeding in order to prevent exsanguinations. A simultaneous effort is made to restore blood volume and correct coagulation. Reversal of coagulopathy with pharmacotherapeutic interventions may be a promising concept to limit blood loss after trauma. Abdominal ultrasound has replaced diagnostic peritoneal lavage for detection of haemoperitoneum. With the development of sliding-gantry based computer tomography diagnostic systems, rapid evaluation by CT-scanning of the trauma patient is possible during resuscitation. The concept of damage control surgery, the staged approach in treatment of severe trauma, has proven to be of vital importance in the treatment of exsanguinating trauma patients and is adopted worldwide. When performing 'blind' transfusion or 'damage control resuscitation', a predetermined fixed ratio of blood components may result in the administration of higher plasma and platelets doses and may improve outcome. The role of thromboelastography and thromboelastometry as point-of-care tests for coagulation in massive blood loss is emerging, providing information about actual clot formation and clot stability, shortly (10min) after the blood sample is taken. Thus, therapy guided by the test results will allow for administration of specific coagulation factors that will be depleted despite administration with fresh frozen plasma during massive transfusion of blood components. PMID- 19135194 TI - Associations between intracranial pressure, intraocular pressure and mean arterial pressure in patients with traumatic and non-traumatic brain injuries. AB - INTRODUCTION: Anatomical proximity of the eye and the intracranial space is a fact but the existence of physiological and pathophysiological relationships between them is elusive. The objective of this study was to explore anatomical and pathophysiological interactions between the eye and the intracranial space and to assess clinical utility of intraocular pressure measurement in estimation of intracranial pressure in patients with brain injuries and to discover how haemodynamic instability could influence these interactions. Controversy surrounds the recent literature concerning this problem and the consensus has not been achieved. MATERIALS AND METHODS: We evaluated the correlation between intracranial pressure and intraocular pressure, intracranial pressure and mean arterial pressure, intraocular pressure and mean arterial pressure in 40 patients with brain injuries initially comatose, admitted to our hospital. All patients required the intracranial pressure monitoring on clinical grounds. Simultaneous recordings of intracranial pressure, intraocular pressure and mean arterial pressure were performed. RESULTS: We calculated both the linear correlation coefficient and the Spearman rank-order correlation coefficient for all three relations. We found significant correlation between intraocular pressure and mean arterial pressure in 63% of the tested population. When the power of the test was increased, by considering only patients with 11 or more observations, this ratio increased to 76%. However, the correlation between intraocular pressure and intracranial pressure, as well as, between intracranial pressure and mean arterial pressure was not significant. CONCLUSIONS: There is no anatomical and pathophysiological basis for the statement that intraocular pressure can be used as an indirect estimator of intracranial pressure. PMID- 19135195 TI - Amylase and lipase measurements in paediatric patients with traumatic pancreatic injuries. AB - INTRODUCTION: Pancreatic injuries occur in up to 10% of paediatric patients who suffer blunt trauma. Initial amylase and lipase measurements have not been helpful as a screening tool to detect pancreatic injuries. However, one primarily adult study suggests that a delayed measurement may be useful. MATERIALS AND METHODS: A retrospective chart review was conducted of patients admitted to a Level I paediatric trauma centre from April 1996 to November 2006 with traumatic pancreatic injuries. RESULTS: The trauma database identified 51 patients with traumatic pancreatic injuries. Inclusion and exclusion criteria were met by 26 patients. Patients with initial amylase and lipase levels measured greater than 2h post-injury were more consistently elevated compared to those patients who had levels measured at 2h or less post-injury. There was a significant association between time of measurement and an increased amylase level (p=0.012). No significant association was found for lipase measurements (p=0.178). DISCUSSION AND CONCLUSIONS: In children with blunt pancreatic injury, elevated serum amylase levels were seen in a significantly higher percentage of patients with initial measurements at greater than 2h post-injury compared to those measured at 2h or less. Lipase measurements demonstrated a similar trend. Delayed amylase and lipase measurements may be helpful to detect pancreatic injuries, but further study is needed. PMID- 19135197 TI - Quinaprilat-eluting stents do not attenuate intimal thickening following stenting in porcine coronary arteries. AB - OBJECTIVE: Vascular injury increases angiotensin-converting enzyme (ACE) activity in the vessel wall, and experimental evidence suggests that high-dose oral ACE inhibition reduces intimal hyperplasia following balloon angioplasty. Local drug delivery can achieve high local concentrations which may be especially efficacious in inhibiting tissue growth following stent implantation. The aim of this study was to evaluate the angiographic and histomorphologic effects of quinaprilat-eluting stents in normal porcine coronary arteries. METHODS: Ten pigs received phosphorylcholine-coated stents in each of the three major coronary arteries: one loaded with 780 microg quinaprilat, one with the solvent and one non-loaded control. Quantitative angiography was performed before and after stenting and at 4 weeks follow-up. At this time point the stented arteries were also analyzed using histology and morphometry. RESULTS: Repeated measures ANOVA yielded significantly smaller angiographic lumen in both quinaprilat and solvent groups: 2.62+/-0.31 and 2.65+/-0.31 mm, respectively versus control: 2.70+/-0.32 mm at follow-up, p<0.05. Histology confirmed this finding with an increment in intimal area (2.5+/-0.86 mm(2)) and thickness (0.57+/-0.29 mm) in the quinaprilat group; versus solvent (1.98+/-0.57 mm(2) 0.4+/-0.26 mm) and controls (1.92+/ 0.50mm(2) and 0.41+/-0.18 mm). CONCLUSION: Quinaprilat-eluting stents do not attenuate neointimal thickening following implantation in normal porcine coronary arteries, but rather show a tendency towards the opposite. PMID- 19135196 TI - Synergistic role of inflammation and insulin resistance as coronary artery disease risk factors in African Americans and Caucasians. AB - The separate roles of inflammation and insulin resistance (IR) in the pathogenesis of cardiovascular disease (CVD) are well recognized. We investigated whether presence of inflammation would modify coronary artery disease (CAD) risk prediction in subjects with or without IR. Insulin, glucose, CRP and fibrinogen levels were determined in 317 Caucasians and 222 African Americans undergoing diagnostic coronary angiography. Extent of CAD was defined by a composite score (0-75). The overall prevalence of IR (HOMA-IR>or=3.0) in Caucasians and African Americans was 32.5% and 22.9%, respectively (P<0.05). The degree of CAD (composite score) was higher in subjects with IR (20.7 vs. 14.5, P=0.014 and 20.1 vs. 13.1, P=0.031 for Caucasians and African Americans, respectively), and in a multiple regression model IR was an independent predictor for CAD in both groups. In both ethnic groups, subjects with a combination of IR and high CRP (>or=3mg/l) had significantly higher composite score compared to those with no IR and low CRP (<3mg/l) (21.2 vs. 13.9, P<0.05 and 20.9 vs. 10.2, P<0.05 for Caucasians and African Americans, respectively). Similarly, the composite score was significantly higher in subjects with IR and high fibrinogen (>or=340 mg/dl) compared to those with no IR and low fibrinogen. In conclusion, elevated levels of inflammatory markers were positively associated with IR. Further, a combination of IR and inflammation resulted in a higher degree of CAD in both Caucasians and African Americans. The results suggest that inflammation may potentiate the cardiovascular risk factor role of IR. PMID- 19135198 TI - Evaluation of the association of genetic variants on the chromosomal loci 9p21.3, 6q25.1, and 2q36.3 with angiographically characterized coronary artery disease. AB - OBJECTIVES: The chromosomal loci 9p21.3, 6q25.1, and 2q36.3, represented by their respective leading variants rs1333049, rs6922269 and rs2943634, have been linked with a history of coronary artery disease (CAD) by genome-wide association studies. Whereas the association of variant rs1333049 with CAD was analysed in several subsequent studies, replication studies of variants rs6922269 and rs2943634 are missing. Furthermore, no direct association with coronary atherosclerosis has been established. We therefore aimed at investigating the association of the above variants with coronary atherosclerosis. METHODS: We performed genotyping in two large cohorts of consecutive Caucasian patients undergoing coronary angiography for the evaluation of suspected or established stable CAD, comprising 671 and 940 patients, respectively, with a total of 1611 subjects. RESULTS: In models of dominant inheritance, variant rs1333049 conferred a significantly increased risk of significant coronary stenoses with lumen narrowing >or=50% in both study cohorts, with adjusted odd ratios (OR) of 1.71 (1.15-2.52); p=0.007 and 1.55 (1.10-2.18); p=0.012, respectively. Variant rs6922269 in neither cohort was significantly associated with CAD. Although carriers of the A allele of variant rs2943634 were at an increased risk of significant coronary stenoses in the second cohort (OR=1.41 (1.06-1.88); p=0.018), no such association was found for the first cohort nor for both cohorts combined. CONCLUSION: Our data from two populations show that variant rs1333049 is significantly associated with angiographically characterized CAD. In contrast, variant rs6922269 did not show any impact on coronary atherosclerosis. The association between variant rs2943634 and CAD warrants further investigation. PMID- 19135200 TI - The effect on patellofemoral joint stability of selective cutting of lateral retinacular and capsular structures. AB - Patient selection for lateral retinacular release (LRR) and its efficacy are controversial. Iatrogenic medial subluxation can occur with inappropriate LRR. The aim of this study was to determine the reduction in patellofemoral stability with progressively more extensive LRR. The force required to displace the patella 10mm medially and laterally in nine cadaveric knees was measured with and without loading of the quadriceps and iliotibial band. The knee was tested intact, then after progressive release beginning proximal to the patella (PR), the mid-level between the proximal and distal limit of the patella (MR) where the fibres are more transverse, then distally till Gerdy's tubercle (DR) and finally the joint capsule (CR). Both medial and lateral stability decreased with progressive releases, larger for the medial. The MR caused a significant reduction of lateral stability between 30 degrees and 90 degrees of knee flexion. There was an 8% reduction in medial stability at 0 degrees flexion with a complete LRR (DR). A comparable reduction in medial stability in the loaded knee at 20 degrees and 30 degrees flexion was obtained with MR alone, with no further reduction after DR. A capsular release caused a further reduction in medial stability at 0 degrees and 20 degrees and this was marked in the unloaded knee. In extension, the main lateral restraint was the joint capsule. At 30 degrees flexion, the transverse fibres were the main contributor to the lateral restraint. PMID- 19135199 TI - Clinical and genetic factors associated with lipoprotein-associated phospholipase A2 in the Framingham Heart Study. AB - OBJECTIVE: To conduct an investigation of clinical and genetic correlates of lipoprotein-associated phospholipase (Lp-PLA(2)) activity and mass in a large community-based cohort. Higher circulating Lp-PLA(2) predicts cardiovascular disease risk, but sources of inter-individual variability are incompletely understood. METHODS: We conducted stepwise regression of clinical correlates of Lp-PLA(2) in four Framingham Heart Study cohorts (n=8185; mean age 50+/-14 years, 53.8% women, 9.8% ethnic/racial minority cohort). We also conducted heritability and linkage analyses in Offspring and Generation 3 cohorts (n=6945). In Offspring cohort participants we performed association analyses (n=1535 unrelated) with 1943 common tagging SNPs in 233 inflammatory candidate genes. RESULTS: Sixteen clinical variables explained 57% of the variability in Lp-PLA(2) activity; covariates associated with Lp-PLA(2) mass were similar but only explained 27% of the variability. Multivariable-adjusted heritability estimates for Lp-PLA(2) activity and mass were 41% and 25%, respectively. A linkage peak was observed for Lp-PLA(2) activity (chromosome 6, LOD score 2.4). None of the SNPs achieved experiment-wide statistical significance, though 12 had q values <0.50, and hence we expect at least 50% of these associations to be true positives. The strongest multivariable-association with Lp-PLA(2) activity was found for MEF2A (rs2033547; nominal p=3.20 x 10(-4)); SNP rs1051931 in PLA2G7 was nominally associated (p=1.26 x 10(-3)). The most significant association to Lp-PLA(2) mass was in VEGFC (rs10520358, p=9.14 x 10(-4)). CONCLUSIONS: Cardiovascular risk factors and genetic variation contribute to variability in Lp-PLA(2) activity and mass. Our genetic association analyses need replication, which will be facilitated by web posting of our genetic association results. PMID- 19135201 TI - A micromechanical model to predict damage and failure in biological tissues. Application to the ligament-to-bone attachment in the human knee joint. AB - Computational models are developed in injury biomechanics to assess lesions in biological tissues based on mechanical measurements. The linear mechanics of fracture theory (LMFT) is a common approach to establish injuries based on thresholds (such as force or strain thresholds) which are straightforward to implement and computationally efficient. However, LMFT does not apply to non linear heterogeneous materials and does not have the ability to predict failure onset. This paper proposes the cohesive zone model theory (CZMT) as an alternative. CZMT focuses on the development of behaviour laws for crack initiation and propagation at an interface that apply within a fibrous material or at the interface between materials. With the view of evaluating CZMT for biological tissues, the model developed by Raous et al. [1999. A consistent model coupling adhesion, friction and unilateral contact. Comput. Methods Appl. Mech. Eng., 177, 383-399] was applied to the ligament-to-bone interface in the human knee joint. This model accounts for adhesion, friction and damage at the interface and provides a smooth transition from total adhesion to complete failure through the intensity of adhesion variable. A 2D finite element model was developed to mimic previous experiments, and the model parameters were determined using a dichotomy method. The model showed good results by its ability to predict damage. The extension to a 3D geometry, with an inverse problem approach, is, however, required to better estimate the model parameters values. Although it is computationally costly, CZMT supplements the improvements achieved in microimaging techniques to support the development of micro/macro approaches in biomechanical modelling. PMID- 19135202 TI - Surface slide track mapping of implants for total disc arthroplasty. AB - Total disc arthroplasty has recently become a potential alternative to spinal arthrodesis. Until recently, there has been no standardized method for evaluating the wear of an artificial disc and myriad testing conditions have been used. The American Society for Testing and Materials (ASTM) and International Organization of Standardization (ISO) recently published guidance documents for the wear assessment of intervertebral spinal disc prostheses; however, various kinematic profiles are suggested, leading to different wear paths between the articulating surfaces of the implants. Since the wear between materials is influenced by the type of relative motion, it is important to select test conditions that lead to clinically realistic results. The purpose of this study was to characterize the slide tracks generated by 7 test conditions allowed for by the ISO and ASTM guidance documents and in Euler sequences consistent with 4 commercially available spine wear simulators. The analysis was performed for a ball-in-socket articulation under both lumbar and cervical motion test conditions. Results were generated analytically using a mathematical algorithm and then validated experimentally. Four tests resulted in elliptical sliding tracks of similar geometries for both the lumbar and cervical conditions. Curvilinear and ribbon shaped wear paths were generated for 3 tests. With the data normalized for implant diameter, the sliding distance was similar between the lumbar and cervical conditions allowed for in the ASTM guidance. This distance differed compared with the results for the ISO guidance document where the lengths of cervical slide tracks were twice those for the lumbar conditions. Slide tracks were also found to be insensitive to the type of simulator under all testing conditions. PMID- 19135203 TI - Strain field in actin filament network in lamellipodia of migrating cells: implication for network reorganization. AB - Cell motility is spatiotemporally regulated by interactions among mechanical and biochemical factors involved in the regulation of cytoskeletal actin structure reorganization. Although the molecular mechanisms underlying cell motility have been well investigated, the contributions of mechanical factors such as strain in the network reorganization remain unclear. In this study, we have quantitatively evaluated the strain field in the actin filament network forming the lamellipodia of migrating fish keratocytes to elucidate the mechanism by which actin filament network reorganization is regulated by biomechanical factors. The results highlight the existence of a negative (compressive) strain in the lamellipodia whose direction is parallel to that of cell movement. A close correlation was found between the distributions of the strain and the actin filament density in the lamellipodia, suggesting that negative strain may be involved in filament depolymerization. Based on this result, we propose a selective depolymerization model which suggests that negative strain may couple with biomechanical factors such as ADF/cofilin to promote selective depolymerization of filaments oriented in the direction of the deformation because such filaments experience relatively higher levels of the deformation. This model, in conjunction with others, may explain the observed reduction in filament density and the reorganization of actin filament network at the back of the lamellipodia of migrating fish keratocytes. Thus, we suggest that by coupling with biochemical factors, mechanical factors are involved in the regulation of actin filament depolymerization, thereby contributing to the regulation of cell motility. PMID- 19135204 TI - An inverse method for predicting tissue-level mechanics from cellular mechanical input. AB - Extracellular matrix (ECM) provides a dynamic three-dimensional structure which translates mechanical stimuli to cells. This local mechanical stimulation may direct biological function including tissue development. Theories describing the role of mechanical regulators hypothesize the cellular response to variations in the external mechanical forces on the ECM. The exact ECM mechanical stimulation required to generate a specific pattern of localized cellular displacement is still unknown. The cell to tissue inverse problem offers an alternative approach to clarify this relationship. Developed for structural dynamics, the inverse dynamics problem translates measurements of local state variables (at the cell level) into an unknown or desired forcing function (at the tissue or ECM level). This paper describes the use of eigenvalues (resonant frequencies), eigenvectors (mode shapes), and dynamic programming to reduce the mathematical order of a simplified cell-tissue system and estimate the ECM mechanical stimulation required for a specified cellular mechanical environment. Finite element and inverse numerical analyses were performed on a simple two-dimensional model to ascertain the effects of weighting parameters and a reduction of analytical modes leading toward a solution. Simulation results indicate that the reduced number of mechanical modes (from 30 to 14 to 7) can adequately reproduce an unknown force time history on an ECM boundary. A representative comparison between cell to tissue (inverse) and tissue to cell (boundary value) modeling illustrates the multiscale applicability of the inverse model. PMID- 19135205 TI - One single standard substance for the determination of multiple anthraquinone derivatives in rhubarb using high-performance liquid chromatography-diode array detection. AB - Due to the complexity of the chemical constitution of traditional Chinese medicines (TCMs), now there is a trend to establish methods, using multi components analysis, for the effective quality control of TCMs. However, the limited availability of multiple reference substances hinders the wide popularization for routine quality control of TCMs. Using an easily available single component contained in the analyte as an internal standard to determine multiple analogues should be a practical option. In this study, we selected rhubarb rhizome as an example, and used emodin, the easily available active component, as the external standard to determine directly the content of emodin in rhubarb, and the same component as the internal standard to simultaneously determine the other six anthraquinones in rhubarb. Compared with the results obtained using the external standard method, this alternative method was found to have no statistically significant differences in our laboratory as verified by the F-test (p=95%, n=6). However, due to weak robustness caused by the fluctuation of relative response factors in different laboratories, such a method can only serve as a practical measure to solve the lack of so many chemical standard substances when relative response factors have been determined. This alternative method can then be used without reference substances. Once the corresponding chemical standards are available and are acceptable as well as cost effective, then the external standard method for the simultaneous determination of multiple components (SDMC) in TCMs will prevail. PMID- 19135206 TI - Challenge of high polarity and low concentrations in analysis of cytostatics and metabolites in wastewater by hydrophilic interaction chromatography/tandem mass spectrometry. AB - A method for solid phase extraction and HPLC-MS/MS of the cytostatics 5 fluorouracil, cytarabine, and gemcitabine and human metabolites uracil 1-beta-d arabinofuranoside and 2',2'-difluorodeoxyuridine in wastewater was established. Wastewater samples from a Swiss hospital were analyzed for 5-fluorouracil, gemcitabine and 2',2'-difluorodeoxyuridine. The limits of quantification were 5.0, 0.9, and 9.0ng/L and the maximum concentrations detected were 27, 38, and 840ng/L, respectively. Along with the method development, retention mechanisms on the hydrophilic interaction chromatography (HILIC) stationary phase were studied. Both partitioning and adsorption play a role in the retention on the tested sulfoalkylbetaine modified silica HILIC column material. The contribution of these two processes is changing over the 1.6-40% range water in the mobile phase. Although the specific break point is difficult to determine, adsorption becomes more significant as the fraction of water in the mobile phase decreases below approximately 16%. PMID- 19135208 TI - Contribution of 1H NMR to the investigation of the adsorption of cationic Gemini surfactants with oligooxyethylene spacer group onto silica. AB - The present study aims to investigate the behavior of a series of cationic Gemini surfactants with a hydrophilic spacer at liquid-gas and solid-liquid interfaces, with particular emphasis on the effect of spacer length. Gemini surfactants containing two quaternary ammonium groups bound by an ethylene oxide spacer chain, referred to as 12-EO(x)-12 with x = 1,3,7 and 12 were synthesized. Surface tension measurements were used to show that the hydrophilic spacer with oxyethylene moieties was not fully extended at the air-water interface. With increasing the spacer group size, it became sufficiently flexible to adopt a particular conformation with a loop at the water side of the interface. A combined study by adsorption isotherm measurements and (1)H NMR spectroscopy allowed a detailed description of the adsorption mechanism of these investigated 12-EO(x)-12 surfactants, with NMR providing more precise information on the conformation of hydrophilic spacer at the solid-liquid interface. Binding to the silica surface involved one cationic headgroup for the surfactants with a short spacer and the two headgroups for the ones with a long spacer. The number of charged surface sites was estimated by considering the dimeric surfactant as a "molecular ruler." The small density of adsorption sites gave rise to the formation of pinned surface micelles. PMID- 19135207 TI - Single-drop microextraction with in-microvial derivatization for the determination of haloacetic acids in water sample by gas chromatography-mass spectrometry. AB - A new approach using single-drop microextraction (SDME) and gas chromatography mass spectrometry for the determination of six haloacetic acids (HAAs) in water samples was presented. n-Octanol was used as extractant and derivatization reagent. HAAs were derivatized both simultaneously during the extraction in the solvent microdrop, and after extraction, inside a glass microvial (1.1mm I.D.). Trifluoroacetic anhydride (TFAA) was used as the reaction catalyst. The influence of catalyst amount, derivatization time and temperature on the yield of the in microvial derivatization was investigated. Derivatization reaction was performed using 1.2microL of TFAA at 100 degrees C for 20min. Extraction was performed using 1.8microL of n-octanol containing TFAA (10%, v/v). Experimental parameters, such as, exposure time, sample pH and extraction temperature were controlled and optimized. Analytical parameters such as linearity, precision and limit of detection were also evaluated. The proposed method was proved to be a suitable analytical procedure for HAAs in water with limits of detection 0.1-1.2microg/L. The relative recoveries range from 82.5 to 97.6% for all the target analytes. Precision values were from 5.1 to 8.5% (as intra-day relative standard deviation, RSD) and 8.8-12.3% (as inter-day RSD). PMID- 19135209 TI - Functionalized gold nanoparticles: synthesis, structure and colloid stability. AB - Gold nanoparticles and their arrays are some of the most studied nanomaterials, with promising applications in many fields such as electronics, optoelectronics, catalysis and biology. In order to protect bare gold nanoparticles from aggregation, to manipulate the optical, electronic and catalytic properties of the gold core, as well as to control interfacial properties, the gold nanoparticles are generally capped by an organic layer. Previous studies [C.D. Bain, G.M. Whitesides, J. Am. Chem. Soc. 110 (1988) 3665-3666] have revealed that many phenomena (e.g., wetting, friction and adhesion), are sensitive to the top few angstroms of a surface. The interfacial properties of a gold surface derivatized with a self-assembled monolayer will thus be dictated by the functionalities present on the outer side of the monolayer. The synthesis, functionalization and surface structure of monolayer-protected gold nanoparticles have been intensively studied in recent times [M.-C. Daniel, D. Astruc, Chem. Rev. 104 (2004) 293-346]. In addition, the aggregation and dispersion of colloidal nanoparticles is one of the key issues related to their potential applications. The forces that govern the colloid stability of nanoparticle dispersions, and how to control them, have yet to be fully investigated. Here special attention has been paid to control of colloid stability using external stimuli. In this feature article, the following five areas are reviewed: synthesis and applications of nanostructured particles; formation and structure of self-assembled monolayer protected gold nanoparticles; colloid stability-DLVO and non-DLVO forces; photochemistry, photochromism and pyrimidine; and manipulation of colloid stability with external stimuli. PMID- 19135210 TI - Effect of different vaccine formulations on the development of Glasser's disease induced in pigs by experimental Haemophilus parasuis infection. AB - Four groups of pigs immunized with different vaccines and a group of non vaccinated controls were challenged intratracheally with a lethal dose (5 x 10(9) colony-forming units) of Haemophilus parasuis, the aetiological agent of Glasser's disease. A vaccine containing inactivated whole organisms gave strong protection against clinical signs, death, pathological changes and persistence of organisms in vivo. However, all non-immunized pigs, all pigs given a vaccine consisting of the recombinant transferring-binding protein (Tbp) B, some pigs given an outer membrane protein (OMP) formulation enriched with TbpB and some pigs immunized with a sub-lethal dose of live organisms died at various times after challenge, yielding positive cultures from most organs post mortem and having shown hyperthermia and other clinical signs before death. Animals that died showed fibrinosuppurative polyserositis, exudative pneumonia, and lesions compatible with acute septicaemia, e.g., disseminated intravascular coagulation with multiple fibrinous thrombi in arterioles and capillaries, depletion of splenic white pulp, and acute lymphadenitis. The results suggested that, in addition to the protection given by inactivated whole organisms, partial protection was given by the OMP formulation and by a sub-lethal dose of living organisms; however, the recombinant TbpB preparation gave no protection. PMID- 19135211 TI - Parvovirus infection in a Eurasian lynx (Lynx lynx) and in a European wildcat (Felis silvestris silvestris). AB - A Eurasian lynx and a European wildcat from the same wildlife park were submitted for necropsy examination after sudden death and after death following a clinical history of lethargy, respectively. Neither animal had been vaccinated against feline parvovirus (feline panleukopenia virus). Feral domestic cats were widespread in the area of the wildlife park and a number of these animals that had been captured had recently died from parvovirus infection. Gross and microscopical findings in the two non-domestic felids were consistent with feline parvovirus infection and this was confirmed by immunohistochemistry and polymerase chain reaction. The introduction of feline parvovirus into captive non domestic felid populations could pose a threat to their health and survival. Vaccination of captive non-domestic felids is therefore recommended. PMID- 19135212 TI - Expression of class II major histocompatibility complex molecules in chronic pulmonary Mycoplasma bovis infection in cattle. AB - Pulmonary inflammation often results in expression of the class II major histocompatibility complex (MHCII) by both professional antigen-presenting cells (APCs; histiocytes and lymphocytes) and non-professional APCs (respiratory epithelium and endothelium). In this study lesions from 17 cases of bovine chronic pneumonia, associated with Mycoplasma bovis infection, were examined immunohistochemically for M. bovis antigen and MHCII expression. Ten cases of chronic necrosuppurative bronchopneumonia (NBP) were shown to be characterized by abundant perinecrotic M. bovis antigen associated with scant MHCII expression by degenerate leucocytes. Seven cases of chronic catarrhal bronchointerstitial pneumonia (CBP) showed prominent MHCII expression by both professional APCs and respiratory epithelium, in the absence of intralesional M. bovis immunolabelling. The results suggest that prominent MHCII expression by both professional and non professional APCs plays a role in the pathogenesis of M. bovis-induced CBP. Conversely, the role of MHCII expression in necrosuppurative foci typical of M. bovis-associated NBP can be considered negligible. PMID- 19135213 TI - No association between APOE and major depressive disorder in a community sample of 17,507 adults. AB - Mood-related phenotypes are commonly comorbid with, and have been implicated in the development of, neurological disorders. APOE is a major susceptibility gene for neurodegeneration. Recent evidence from case-control studies has suggested that the apoE 2 allele is associated with major depressive disorder (MDD). However, evidence from large-scale community-based studies is limited. APOE was genotyped for 17,507 men and women, aged 41-80 years, participating in the European Prospective Investigation into Cancer-Norfolk study, who had also completed a psychosocial assessment that included measures of emotional health status defined by MDD, psychological distress (as represented by the Mental Health Inventory, MHI-5), and by an assessment of neuroticism. No associations were found between APOE genotypes and measures either of past-year or lifetime MDD, or of emotional health defined according to the MHI-5 or by neuroticism. Data from this large-scale, community-based, study are not supportive of an association between either MDD or associated measures of emotional state and APOE genotype. These findings suggest that the association between APOE and MDD risk is more modest than has been previously reported. PMID- 19135214 TI - Low serum cholesterol and external-cause mortality: potential implications for research and surveillance. AB - OBJECTIVE: Previous studies suggested that low total cholesterol was associated with external mortality, including deaths from suicide, homicide, and accidents. However, this reported association was potentially confounded, since cholesterol was also reported to be associated with alcohol abuse, anti-social personality disorder, and other risk factors for external mortality. METHOD: We examined external-cause mortality among a national sample of 4462 male, US veterans at baseline in 1985. Using Cox regressions to estimate survival time, we assessed the impact of low baseline total cholesterol < or =165 mg/dl, age, race, intelligence, BMI, alcohol abuse, anti-social personality disorder, depression, and other factors at follow-up. Study follow-up continued until December 31, 2000. A total of 55 external mortalities occurred during this approximately 16 year period. RESULTS: Multivariate Cox regressions predicting external-cause mortality suggested that three predictor variables were significant: low total cholesterol, morbid depression, and anti-social personality disorder, with hazard ratios (HRs) of 1.97 (p=0.046), 1.76 (p=0.043), and 2.22 (p=0.006), respectively. In addition, a significant interaction was detected for low cholesterol x morbid depression (p<0.005), whereby those with both at baseline were approximately 7 times more likely to die from external mortality (HR=6.5, 95% CI=3.07-13.76). CONCLUSION: Among a national random sample of community-based men, lower baseline cholesterol predicted external mortality and revealed an interaction with morbid depression. Patients presenting with low cholesterol and morbid depression in clinical practice may warrant clinical attention and surveillance. PMID- 19135215 TI - Differential responses in CBF and CBV to cocaine as measured by fMRI: implications for pharmacological MRI signals derived oxygen metabolism assessment. AB - INTRODUCTION: Cognitive performance-induced brain oxygen metabolism has been successfully measured by functional magnetic resonance imaging (fMRI) in human studies. The measurement of the cerebral metabolic rate of oxygen consumption (CMRO2) is typically achieved by assuming a fixed coupling of cerebral blood flow (CBF) and cerebral blood volume (CBV) and by performing a separate experiment to assess the vascular response to a hypercapnic challenge. Psychoactive drugs may have directly effect on the cerebral vasculature, potentially confounding the interpretation of pharmacological MRI (phMRI) data. In this study, we tested the assumptions of the standard CMRO2 calculation following the administration of cocaine, in order to test the validity of this measurement in phMRI studies. The initial transient state and later steady state CBF and CBV responses to a hypercapnic challenge were measured. METHODS: CBF and CBV responses were directly measured by fMRI using continuous arterial spin-labeling (ASL) and contrast enhanced fMRI, respectively. The coupling between changes in CBF and CBV during a hypercapnic challenge was examined under normal conditions and following the administration of cocaine. RESULTS: A decoupling of changes in CBF and CBV was observed during the transient state immediately following the administration of cocaine, and an altered coupling of CBF and CBV was found during the steady state after cocaine injection. DISCUSSION: These data suggest caution in interpreting CMRO2 measurements from phMRI studies and may also lead to an improved understanding of the complex neuronal and vascular mechanisms of drug action. PMID- 19135216 TI - Primary diffuse leptomeningeal gliomatosis mimicking a chronic inflammatory meningitis. AB - Primary diffuse leptomeningeal gliomatosis (PDLG) is a rare, fatal, neoplastic condition of infiltrating glial cells into the meninges without evidence of primary tumor in the brain or spinal cord parenchyma. Primary diffuse leptomeningeal gliomatosis often presents with symptoms and physical findings of chronic inflammatory meningitis and raised intracranial pressure, and lacks specific clinical, radiologic, and diagnostic criteria. We report a case of PDLG diagnosed post-mortem, highlighting the diagnostic difficulty in identifying PDLG as the cause of chronic meningitis, even when a neoplastic etiology is suspected. Because multiple cytologies and even a leptomeningeal biopsy did not reveal the diagnosis ante-mortem, we emphasize the consideration of multi-site or repeat leptomeningeal biopsy when a persistent inflammatory infiltrate is found and neurological symptoms are progressive. PMID- 19135217 TI - Isolated labyrinthine infarction as a harbinger of anterior inferior cerebellar artery territory infarction with normal diffusion-weighted brain MRI. AB - OBJECTIVES: This paper aims to determine the frequency of isolated labyrinthine infarction as an initial manifestation of anterior inferior cerebellar artery (AICA) territory infarction and discuss its clinical implication. METHODS: We studied 54 consecutive patients with AICA infarction diagnosed by brain MRI from the acute stroke registry. RESULTS: We identified 4 patients (7.4%) with AICA territory infarction who initially presented with vertigo and hearing loss mimicking acute labyrinthitis and subsequently suffered from delayed neurological deficits. All patients had normal brain MRIs including diffusion-weighted images at the time of isolated audiovestibular loss, but all subsequently exhibited acute infarct(s) in the AICA territory on follow-up brain MRIs when they developed additional neurological deficits. CONCLUSION: Labyrinthine infarction may be a warning sign of impending pontocerebellar infarction in the AICA territory. Our results indicate that even a classical peripheral presentation, e.g., acute hearing loss, and acute vertigo with unidirectional nystagmus, may be a manifestation of ischemic stroke in the posterior circulation. PMID- 19135218 TI - Baseline metal concentrations in marine algae from Sao Miguel (Azores) under different ecological conditions--urban proximity and shallow water hydrothermal activity. PMID- 19135219 TI - The Endo-Distractor for preimplant mandibular regeneration. AB - OBJECTIVE: A simple practical device for a new technique of vertical distraction osteogenesis was developed. The Endo-Distractor Krenkel was originally intended for the anterior regeneration of highly atrophic mandibles. The Endo-Distractor features several novelties. Placement is made intraosseously in the basal cortical bone. The distraction screw length may be adapted to the depth of chin soft tissues. The quality of anchorage in the basal cortical bone guarantees the stability of the distraction vector. The endobuccal emergence is that of an artificial crown, and does not cause any patient discomfort. The device can easily be removed without secondary surgery. All kinds of implants may be placed after the retention time. This study's objective was to evaluate the use of the Endo-Distractor Krenkel in edentulous patients with highly atrophic mandibles. MATERIAL AND METHOD: This new device was used on 18 patients, between January 2000 and September 2004, who were then followed-up for at least 36 months. Mandibular atrophy was measured with a lateral cephalogram, then classified according to Atwood's modified classification. The studied criteria included the amplitude of distraction, its duration, the distractor's lingual tilt, the number and outcome of implants, and complications. RESULTS: The sex ratio was 17 female for one male patient. The mean patient age was 56 years (43 to 66 years). The mean distraction amplitude was 11.3mm (8 to 14 mm). The mean retention time was 186.8 days (37 to 309 days). The distractor's mean lingual tilt was 4.3 degrees (0 to 23 degrees ). Two mandibular fractures occurred 6 weeks after placing the Endo-Distractor. The first one was treated medically, and the second one required removing the Endo-Distractor and osteosynthesis. An average of four interforaminal implants were placed for a total of 24 Branemark and 51 Straumann implants. Four implants were lost in a patient due to infection. All other implants were osseointegrated. No bone loss was detected at follow-up after functional loading. DISCUSSION: These results show that alveolar distraction is possible on severely atrophic mandibles. The quality of bone and gum reconstruction is satisfactory both for functional and esthetic results. Surgical difficulty and rate of complications were lower than with conventional distraction techniques. PMID- 19135220 TI - Schistosomal peritonitis: a rare cause of acute abdomen. AB - Schistosomiasis is a major parasitic infection of tropical areas. We present a 26 year-old male immigrant from Egypt with acute abdomen due to schistosomial infection, mimicking symptoms of acute appendicitis, with histopathological detection and representation of schistosomial eggs in the unusual area of the greater omentum. This is a rare case with findings of intra-peritoneal schistosoma ova. We review the pertinent literature and indicate the need for a new approach to infectious diseases with unusual clinical presentation, due to pathogens coming from distal geographical regions, in which surgeons must keep a high index of suspicion. PMID- 19135221 TI - Distributions of the endocrine cells in the gastrointestinal tract of nectarivorous and sanguivorous bats: a comparative immunocytochemical study. AB - The present study was conducted to clarify the influence of feeding habits on regional distribution and relative frequency of endocrine cells secreting cholecystokinin (CCK), gastrin (GAS), serotonin (5-HT) and enteroglucagon (GLUC) in the nectarivorous Anoura geoffroyi and Glossophaga soricina and the sanguivorous Desmodus rotundus bats of the Phyllostomidae family, by specific immunohistochemical methods. The regional distribution and frequency of the different types of endocrine cells varied according to their location in the GIT. 5-HT immunoreactive cells (IR), detected throughout the GIT of three bats, were the most predominant gastrointestinal endocrine cells. GAS-IR cells in A. geoffroyi were found at the base of the pyloric gland, while in G. soricina they could also be observed in the middle to basal portions of the gland. GLUC-IR cells were located in the fundic region of A. geoffroyi, G. soricina and D. rotundus. These endocrine cells were more abundant in the sanguivorous bat. In nectarivorous bats were compared to sanguivorous bat, which differ in dietary habits, difference in the distribution and relative frequency of gut endocrine cells would be predicted. The absence of some, and decrease in frequency of other, gastrointestinal endocrine cells may reflect, in part, its interspecies differences or dietary habits. PMID- 19135222 TI - Papillomaviruses in the causation of human cancers - a brief historical account. AB - Approximately 35 years ago a role of human papillomaviruses (HPV) in cervical cancer has been postulated. Today it is well established that this very heterogeneous virus family harbours important human carcinogens, causing not only the vast majority of cervical, but also a substantial proportion of other anogenital and head and neck cancers. In addition, specific types have been linked to certain cutaneous cancers. In females, HPV infections on a global scale account for more than 50% of infection-linked cancers, in males for barely 5%. Vaccines against the high risk HPV types 16 and 18 represent the first preventive vaccines directly developed to protect against a major human cancer (cervical carcinoma). This review will cover some of the historical aspects of papillomavirus research; it tries briefly to analyze the present state of linking HPV to human cancers and will discuss some emerging developments. PMID- 19135223 TI - Biochemical and biophysical comparison of cleaved and uncleaved soluble, trimeric HIV-1 envelope glycoproteins. AB - Human immunodeficiency virus type 1 (HIV-1) entry into host cells is mediated by the trimeric envelope glycoprotein complex (Env). Accordingly, the Env proteins are the targets for neutralizing antibodies (NAbs) and are the focus of vaccines intended to induce NAbs. Because the Env complex is labile, soluble recombinant Env (gp140) trimers require engineering to stabilize them sufficiently for use as immunogens. Trimeric forms of gp140 trimers can be created that are either cleavage-competent or cleavage-defective at the junction between the gp120 and gp41 subunits. As functional trimers are cleaved at this site, the question arises as to whether cleavage affects the antigenic structure of the Env complex in a way that is relevant to vaccine design. Here, we present a comparative analysis of the antigenicity profiles of cleaved and uncleaved gp140 trimers derived from the KNH1144 (subtype A) virus that are otherwise closely sequence matched. While cleavage did not affect the exposure of NAb epitopes on the gp140 trimers, non-neutralizing antibodies to gp41 epitopes bound much more strongly to uncleaved trimers. Hence cleavage does alter the structure of the HIV-1 Env complex. PMID- 19135224 TI - Expression of the human papillomavirus type 16 E7 oncoprotein induces an autophagy-related process and sensitizes normal human keratinocytes to cell death in response to growth factor deprivation. AB - Expression of oncogenes, such as the human papillomavirus type 16 (HPV16) E7 oncoprotein, promotes aberrant cell proliferation. In the absence of concurrent mitogenic stimuli, this triggers a cell-intrinsic defense mechanism, the "trophic sentinel response", which eliminates such aberrant cells. The molecular pathways that elicit this response, however, remain obscure. We set up an experimental system to investigate the trophic sentinel pathway triggered by HPV16 E7 expression in normal human keratinocytes, the natural host cells of HPVs. Keratinocytes expressing HPV16 E7 cultured in E-medium undergo cell death and show increased sub-G1 DNA content when grown to confluence or under conditions of serum deprivation. Moreover, HPV16 E7 expressing human keratinocytes express higher levels of the autophagy marker, LC3-II, which can be abrogated by 3 methyladenine, an autophagy inhibitor. These findings indicate that even under normal culture conditions, HPV16 E7 expression triggers metabolic stress that may result in autophagy, a pathway implicated in carcinogenesis. PMID- 19135226 TI - Identification of the immediate-early genes of white spot syndrome virus. AB - During viral infection, viral immediate-early (IE) genes encode regulatory proteins critical for the viral life cycle. Here we screened white spot syndrome virus (WSSV) IE genes with cycloheximide (CHX)-treated primary culture of crayfish hemocyte and a WSSV genome tiling microarray. Sixteen ORFs, including a known WSSV IE gene (ie1/wsv069), were identified and confirmed by RT-PCR and time course studies. The 16 identified IE proteins contain four proteins (wsv051, wsv069, wsv100, wsv079) with transcription activity, one (wsv083) with Ser/Thr kinase domain and one (wsv249) previously described to function as an ubiquitin E3 ligase. Furthermore, most of the identified WSSV IE genes cluster in a 14 kb genomic region (WSSV China isolate: 36,052 to 50,300 bp). This type of arrangement may facilitate the coordinate control and rapid expression of IE genes. PMID- 19135225 TI - Increased expression of suppressor of cytokine signaling-1 (SOCS-1): A mechanism for dysregulated T helper-1 responses in HIV-1 disease. AB - Maintenance of Th1 responses and dendritic cell (DC) functions are compromised in HIV-1 infected individuals. To better understand these immune abnormalities, we developed an HIV-1 transgenic (Tg) rat. We report that Tg DCs induce elevated levels of SOCS-1 and secrete decreased IL-12p40 and elevated levels of IL-10 following TLR-4 stimulation by LPS. This leads to further induction of SOCS-1 by IL-10 and decreased IFN-gamma-mediated induction of interferon response factor (IRF)-1 and IL-12Rbeta1 expression in CD4+ T cells and to decreased IL-12 induction of IFN-gamma production by Th1 polarized T cells. We also show that SOCS-1 is elevated in CD4+ T cells from HIV-1 infected progressors, and is correlated with defective induction of IRF-1 following IFN-gamma stimulation, compared with healthy controls and HIV-1 natural viral suppressor (NVS) patients. These results suggest a link between high levels of SOCS-1, defects in innate immunity and adaptive Th1 responses that may be reflected in the loss of Th1 immune competence observed with AIDS patients. PMID- 19135227 TI - Electrochemical process combined with UV light irradiation for synergistic degradation of ammonia in chloride-containing solutions. AB - An electrochemical process combined with ultraviolet light irradiation (UPE) using nonphotoactive dimensionally stable anodes (DSAs) like RuO2/Ti and IrO2/Ti in the presence of chlorides was investigated for ammonia degradation. In this process, the in situ electrogenerated active chlorine and in situ photogenerated chlorine radicals were responsible for the high efficiency of ammonia degradation. More than 97% of ammonia was converted to nitrogen and a significantly synergistic effect was confirmed. Compared with the single electrochemical (E) and photochemical (P) process, the degradation rates of ammonia and the average current efficiencies (ACEs) of the UPE process increased by 1.5 and 1.7 times using RuO2/Ti and IrO2/Ti electrodes, respectively. On the basis of the linear voltammograms, Electrochemical Impedance Spectra (EIS), UV vis spectra, Electron Spin Resonance (ESR) analysis and a series of experiments designed, the synergistic mechanism was investigated. In addition, this unique process succeeded in transferring the reaction from the electrode surface to the bulk of the solution compared with the conventional photoelectrocatalytic (PEC) process. The loss of chloride decreased from 21.0% to 7.2% and the recycle of chloride was accelerated in the UPE process. Finally the effects of initial pH, current density and ammonia-nitrogen concentration were discussed. Results indicated that pH and ammonia concentration exerted little influences on the degradation rates and current density was the "rate-determining" factor. PMID- 19135228 TI - Spatial and temporal variations in the occurrence of cocaine and benzoylecgonine in waste- and surface water from Belgium and removal during wastewater treatment. AB - Estimates of cocaine consumption are currently resulting from population surveys, consumer interviews and crime statistics. A new approach ("sewage epidemiology") based on the analysis of cocaine (COC) and its metabolite, benzoylecgonine (BE), in water samples was applied to 10 river sites and 30 wastewater treatment plants (WWTPs) in Belgium. Each river site was sampled twice, during the summer of 2007 and the winter of 2007-2008, while each WWTP was sampled on a Sunday and a Wednesday, during the summer-autumn of 2007 and the winter of 2007-2008. This sampling strategy allowed for the evaluation of spatial and seasonal variations in the occurrence of COC and BE in waste- and surface water. WWTP Brussel-Noord was sampled for 19 consecutive days to evaluate daily and weekly variations in the presence of COC and BE in wastewater. For 7 WWTPs, influent and effluent water samples were collected to investigate the removal of COC and BE during the wastewater treatment process. Analysis of water samples was performed using solid phase extraction and liquid chromatography coupled to tandem mass spectrometry. Measured concentrations were further converted into an amount of used cocaine, called cocaine equivalents, as previously described in the literature. Results showed no significant difference in cocaine use between the investigated seasons. A constant cocaine consumption was observed during the week (Monday-Friday) with peaks during the weekend for WWTP Brussel-Noord. The COC/BE ratio in water samples was significantly higher during winter, most probably due to a slower hydrolysis of cocaine in low-temperature water. COC and BE were removed in the investigated WWTPs with a removal efficiency of >93%. PMID- 19135229 TI - Ground water geochemistry of Ballia district, Uttar Pradesh, India and mechanism of arsenic release. AB - Threat to human health worldwide due to the natural contamination of arsenic in ground waters has led to extensive studies on factors controlling the distribution of arsenic and conditions leading to arsenic mobilization in different arsenic contaminated areas. Another aspect of the arsenic crisis, especially in South Asia, is the degree of spatial variability of ground water arsenic concentrations. Thus it becomes necessary to study the source and the processes involved in arsenic mobilization into ground water under such conditions. An arsenic contaminated area namely, Ballia district of UP was chosen for this study. A set of 56 samples were collected from India Mark II hand pumps (30-33 m depth) thrice in a year namely pre-monsoon (April '07), monsoon (July '06) and winter seasons (December '06). Nine samples were also collected from deep bore well hand pumps (66-75 m) to study the difference in geochemistry with the shallow pumps. Various water quality parameters like As(III), As(V), sulfate, nitrate, phosphate, bicarbonate, ammonia, were determined. Arsenic concentrations ranged from 0 to 468 microg L(-1) in ground water collected from depths of 30-33 m. In the deeper wells (66-75 m), arsenic concentrations ranged from 12 to 20 microg L(-1). Most samples contained both As(III) and As(V) and the concentration of As(III) was generally equal/higher than As(V). Not much variation of arsenic concentration was observed when sampled in summer, monsoon and winter seasons. Correlation studies among various water quality parameters revealed that reductive dissolution of FeOOH was the most probable mechanism for release of arsenic. PMID- 19135230 TI - Atmospheric metal and phosphorus concentrations, inputs, and their biogeochemical significances in the Japan/East Sea. AB - Atmospheric metals and phosphorus over the southern Japan/East Sea were investigated in order to evaluate their sources, concentrations and inputs, and to identify their biogeochemical roles in this marginal sea. Aerosols were collected on the east coast of Korea from February 2002 to April 2003 (n=101) as well as at a remote island (Ulleung) and on a ship from February 2002 to June 2003 (n=13). The aerosols were analyzed for Al, Co, Cu, Ni, P, Pb and Zn. Simultaneous collections of aerosols at both coast and offshore were performed, and several high dust aerosols (Al>5 microg m(-3)) were collected at both regions. At the coastal site, both dust mineral and pollutants were transported by westerly winds from the Asian continent, but local emissions were significant (e.g., Cu, Ni, P and Zn) as well during the summer monsoon (May-August). The experimental relationships between the coast and offshore sites were defined. From these relationships, it was possible to obtain the annually averaged atmospheric metal and P concentrations over the southern Japan/East Sea, which has increased by over 2 times for the last decade. Through the estimation of atmospheric metal and phosphorus fluxes and comparisons with inputs from the Tsushima Warm Current, the atmospheric pathway was found to be a significant source for Al, Pb and Zn. PMID- 19135231 TI - Effect of VKORC1-1639 G>A polymorphism, body weight, age, and serum albumin alterations on warfarin response in Japanese patients. AB - INTRODUCTION: To establish individualized warfarin therapy, we investigated the contribution of genetic variations of vitamin K epoxide reductase complex subunit 1 gene (VKORC1) -1639 G>A and Cytochrome P450 2C9 gene (CYP2C9) and clinical factors on warfarin sensitivity in Japanese patients. MATERIALS AND METHODS: Genetic analyses of VKORC1 -1639 G>A and CYP2C9 2, 3, and 4 were performed in 259 Japanese patients and 341 healthy subjects. We selected 259 patients who have been prescribed warfarin with a 1.5-3.0 range of prothrombin time normalized as an international normalized ratio for at least 3 months and investigated factors that contribute to individual variability in warfarin dose. Furthermore, multivariate analysis was performed to investigate a warfarin dosing algorithm. RESULTS AND CONCLUSIONS: There were great inter-individual differences in warfarin maintenance dose in 259 patients, ranging from a minimum dose of 0.75 mg/day to a maximal dose of 8.00 mg/day. VKORC1 -1639 G>A polymorphism, body weight, age, and serum albumin were found to affect the inter-individual variability. The dosing algorithm of warfarin maintenance dose was investigated by multivariate linear regression. The regression equation was able to account for 33.2% (R(2)(Adj)=0.332) of the overall variability in warfarin dose. PMID- 19135232 TI - Sequential chemotherapy and irradiation in advanced stage endometrial cancer: A Gynecologic Oncology Group phase I trial of doxorubicin-cisplatin followed by whole abdomen irradiation. AB - OBJECTIVES: The optimal treatment regimen for advanced stage endometrial carcinoma (EC) is yet to be established. The objective of this study was to evaluate the feasibility and toxicity of delivering sequential doxorubicin cisplatin (AC) followed by whole abdomen irradiation (WAI). METHODS: Patients with stage III/IV EC with <2 cm residual disease were eligible. The treatment regimen included 3 cycles of AC (50 mg/m(2) each) followed by WAI plus para aortic and/or true pelvic boost. Outcome objectives were feasibility of the regimen, acute toxicity and chronic irradiation toxicity monitored for at least one year after completing treatment. RESULTS: Thirty-one patients were entered onto the study. Twenty-nine were evaluable for feasibility of the regimen, and 22 patients were evaluable for chronic radiation toxicity with a median follow-up of 21 months. Three patients (14%) experienced severe chronic toxicity including one treatment-related death. The five-year progression-free survival (PFS) and overall survival (OS) was 52.5% and 60.1% respectively. CONCLUSIONS: The results from the first stage of this study found this regimen of sequential AC followed by WAI eligible for further study. Although toxicity was not excessive at the time of completion of the first stage, there was insufficient interest by the Gynecologic Oncology Group to continue study of this regimen; therefore, a conclusion regarding feasibility and chronic toxicity in an expanded cohort as initially intended is not possible. The risk of recurrence in advanced EC is sufficiently high that may warrant further investigation of sequential chemotherapy and WAI. PMID- 19135233 TI - Improved detection of ovarian cancer metastases by intraoperative quantitative fluorescence protease imaging in a pre-clinical model. AB - OBJECTIVES: Cytoreductive surgery is a cornerstone of therapy in metastatic ovarian cancer. While conventional white light (WL) inspection detects many obvious tumor foci, careful histologic comparison has shown considerable miss rates for smaller foci. The goal of this study was to compare tumor detection using WL versus near infrared (NIR) imaging with a protease activatable probe, as well as to evaluate the ability to quantify NIR fluorescence using a novel quantitative optical imaging system. METHODS: A murine model for peritoneal carcinomatosis was generated and metastatic foci were imaged using WL and NIR imaging following the i.v. administration of the protease activatable probe ProSense750. The presence of tumor was confirmed by histology. Additionally, the ability to account for variations in fluorescence signal intensity due to changes in distance between the catheter and target lesion during laparoscopic procedures was evaluated. RESULTS: NIR imaging with a ProSense750 significantly improved upon the target-to-background ratios (TBRs) of tumor foci in comparison to WL imaging (minimum improvement was approximately 3.5 fold). Based on 52 histologically validated samples, the sensitivity for WL imaging was 69%, while the sensitivity for NIR imaging was 100%. The effects of intraoperative distance changes upon fluorescence intensity were corrected in realtime, resulting in a decrease from 89% to 5% in signal variance during fluorescence laparoscopy. CONCLUSIONS: With its molecular specificity, low background autofluorescence, high TBRs, and quantitative signal, optical imaging with NIR protease activatable probes greatly improves upon the intraoperative detection of ovarian cancer metastases. PMID- 19135234 TI - Prenatal probiotic administration can influence Bifidobacterium microbiota development in infants at high risk of allergy. PMID- 19135235 TI - Probiotics prevent IgE-associated allergy until age 5 years in cesarean-delivered children but not in the total cohort. AB - BACKGROUND: Less microbial exposure in early childhood is associated with more allergic disease later. Allergic children have a different fecal microflora, with less lactobacilli and bifidobacteria. Beneficial effects regarding the development of allergy have been suggested to come through probiotic supplementation. OBJECTIVE: We sought to study the effect of probiotic and prebiotic supplementation in preventing allergies. METHODS: In a double-blinded, placebo-controlled study we randomized 1223 mothers with infants at high risk for allergy to receive a probiotic mixture (2 lactobacilli, bifidobacteria, and propionibacteria) or placebo during the last month of pregnancy and their infants to receive it from birth until age 6 months. Infants also received a prebiotic galacto-oligosaccharide or placebo. At 5 years, we evaluated the cumulative incidence of allergic diseases (eczema, food allergy, allergic rhinitis, and asthma) and IgE sensitization. RESULTS: Of the 1018 intent-to-treat infants, 891 (88%) attended the 5-year visit. Frequencies of allergic and IgE-associated allergic disease and sensitization in the probiotic and placebo groups were similar: 52.6% versus 54.9% and 29.5% versus 26.6%, respectively, and 41.3% in both. No significant difference appeared in frequencies of eczema (39.3% vs 43.3%), atopic eczema (24.0% vs 25.1%), allergic rhinitis (20.7% vs 19.1%), or asthma (13.0% vs 14.1%) between groups. However, less IgE-associated allergic disease occurred in cesarean-delivered children receiving probiotics (24.3% vs 40.5%; odds ratio, 0.47; 95% CI, 0.23% to 0.96%; P = .035). CONCLUSIONS: No allergy-preventive effect that extended to age 5 years was achieved with perinatal supplementation of probiotic bacteria to high-risk mothers and children. It conferred protection only to cesarean-delivered children. PMID- 19135236 TI - Local release of B cell-activating factor of the TNF family after segmental allergen challenge of allergic subjects. AB - BACKGROUND: Local production of IgA and IgE in the airways has been proposed to be an important event in both immune protection from pathogens and the pathogenesis of airway allergic diseases. OBJECTIVE: The objective of this study was to investigate the production of B cell-activating factor of the TNF family (BAFF), an important regulator of B-cell survival and immunoglobulin class-switch recombination, in bronchoalveolar lavage (BAL) fluid after segmental allergen challenge of allergic subjects. METHODS: Segmental allergen challenge with saline or allergen was performed in 16 adult allergic subjects. BAL was performed at both saline- and allergen-challenged sites 20 to 24 hours after challenge. Concentrations of B cell-active cytokines, including BAFF, IL-6, and IL-13, were measured by using specific ELISA and cytometric bead array assays. RESULTS: Levels of BAFF protein were significantly increased in BAL fluid after allergen challenge (53.8 pg/mL [range, 0-407.4 pg/mL], P = .001) compared with those at saline-challenged sites (0 pg/mL [0-34.7 pg/mL]). In the BAL fluid after allergen challenge, BAFF levels were significantly correlated with absolute numbers of total cells (r = 0.779, P < .001), lymphocytes (r = 0.842, P < .001), neutrophils (r = 0.809, P < .001), and eosinophils (r = 0.621, P = .010) but did not correlate with macrophages. Normalization to albumin indicated that BAFF production occurred locally in the airways. BAFF levels were also significantly correlated with the other B cell-activating cytokines IL-6 (r = 0.875, P < .001) and IL-13 (r = 0.812, P < .001). CONCLUSION: The antigen-induced production of BAFF in the airway might contribute to local class-switch recombination and immunoglobulin synthesis by B cells. PMID- 19135237 TI - Is structured allergy history sufficient when assessing patients with asthma and rhinitis in general practice? AB - BACKGROUND: Many United Kingdom patients with asthma and rhinitis are allergic, but in primary care few diagnostic and management decisions are made with formal allergy assessment. Arguably, knowing a patient's atopic status might be helpful in distinguishing the cause of disease and in selecting appropriate treatments. OBJECTIVES: Our objective was to estimate the extent to which a formal allergy assessment (a structured allergy history and skin prick tests to 5 common aeroallergens) would improve the precision of allergy diagnosis compared with a patient's self-report or the structured allergy history alone. METHODS: One hundred twenty-seven patients with asthma, rhinitis, or both were recruited from 4 general practices in Wessex, United Kingdom. Allergy status based on the patient's opinion and on structured allergy history alone was compared with formal allergy assessment. Assessments were validated by an independent allergy specialist reviewing the files. Patients were given written advice specific to their allergies and followed up 3 months later to assess satisfaction, recall, and effect on health and behavior. RESULTS: Self-reporting misclassified allergic status in many patients. A structured allergy history alone was little better and resulted in false-positive rates for cat allergy of 32%, grass pollen of 48%, house dust mite of 75%, tree pollen of 54%, and dog of 27% compared with formal allergy assessment. Skin prick testing combined with a structured history was essential to reach a correct causative diagnosis. Three months later, 41% patients had made changes to lifestyle, medications, or both, and 18% reported clinical improvement. CONCLUSIONS: Skin prick testing improves the accuracy of an assessment of allergic status based on patient opinion or a structured allergy history alone. PMID- 19135238 TI - Adiposity, serum lipid levels, and allergic sensitization in Chinese men and women. AB - BACKGROUND: Obesity and allergic diseases have increased dramatically in recent decades. Although adiposity has been associated with asthma, associations with allergic sensitization have been inconsistent. OBJECTIVE: To examine the association of adiposity and lipid profiles with allergic sensitization. METHODS: This study included 1187 rural Chinese twins (653 men) age 18 to 39 years, with skin prick tests, anthropometric and dual-energy x-ray absorptiometry-assessed adiposity measures, and lipid assessments. Allergic sensitization was defined as positive SPT to >/=1 allergen (9 foods and 5 aeroallergens tested). We applied sex-stratified generalized estimating equations to assess the association of adiposity and serum lipids with allergic sensitization, and structural equation models to estimate the genetic/environmental influences on any observed associations. RESULTS: Men had lower percent body fat (% BF) (13.9% vs. 28.8%) but higher rates of allergic sensitization (56.2% vs 36.7%) than women. Men in the highest %BF quartile were 2.1 times more likely to be sensitized than the lowest quartile (95% CI, 1.3-3.5; P trend = .003). In men, the risk of allergic sensitization increased with high-density lipoprotein (HDL) <40 mg/dL (odds ratio = 4.0; 95% CI, 1.8-9.2) and higher low-density lipoprotein quartiles (P trend = .007). This appeared to be partially explained by shared genetic factors between serum lipid levels and allergic sensitization. In females, lower HDL was associated with increased risk of allergic sensitization. CONCLUSION: In this relatively lean Chinese population, higher %BF, lower HDL and higher LDL were associated with greater risk of allergic sensitization, most notable in men. The observed associations among adiposity, serum lipids, and allergic sensitization in men appear to be partially explained by common genetic influences on these traits. PMID- 19135239 TI - Prostatic kallikrein: a new major dog allergen. AB - BACKGROUND: Dog dander is an important cause of respiratory allergy, but the spectrum of known dog allergens appears incomplete. Two lipocalins, Can f 1 and Can f 2, and serum albumin, Can f 3, have been characterized in detail but do not fully account for the IgE antibody-binding activity of dog dander extract. Allergen activity has previously been detected in dog urine but not further characterized. OBJECTIVE: We sought to identify, characterize, and assess the importance of allergen components in dog urine. METHODS: Dog urine was fractionated by means of size exclusion chromatography and examined for IgE antibody binding. A protein present in one fraction displaying IgE antibody binding activity was identified by means of N-terminal sequencing and mass spectrometry. A recombinant form of the protein was produced in Pichia pastoris. IgE antibody binding to dog allergen components among sera of 37 subjects with dog allergy was determined by means of ImmunoCAP analysis. RESULTS: An IgE antibody-binding protein was isolated from dog urine and identified as prostatic kallikrein. A closely related or identical protein was detected in dog dander. The recombinant prostatic kallikrein displayed immunologic and biochemical properties similar to those of the natural protein and bound IgE antibodies from 26 (70%) of 37 sera of subjects with dog allergy, 14 of which reacted to none of Can f 1, Can f 2, or Can f 3. The dog allergen identified here was found to cross react with human prostate-specific antigen, a key culprit in IgE-mediated vaginal reactions to semen. CONCLUSION: Prostatic kallikrein is a new major dog allergen. PMID- 19135240 TI - Regulation of PKD by the MAPK p38delta in insulin secretion and glucose homeostasis. AB - Dysfunction and loss of insulin-producing pancreatic beta cells represent hallmarks of diabetes mellitus. Here, we show that mice lacking the mitogen activated protein kinase (MAPK) p38delta display improved glucose tolerance due to enhanced insulin secretion from pancreatic beta cells. Deletion of p38delta results in pronounced activation of protein kinase D (PKD), the latter of which we have identified as a pivotal regulator of stimulated insulin exocytosis. p38delta catalyzes an inhibitory phosphorylation of PKD1, thereby attenuating stimulated insulin secretion. In addition, p38delta null mice are protected against high-fat-feeding-induced insulin resistance and oxidative stress-mediated beta cell failure. Inhibition of PKD1 reverses enhanced insulin secretion from p38delta-deficient islets and glucose tolerance in p38delta null mice as well as their susceptibility to oxidative stress. In conclusion, the p38delta-PKD pathway integrates regulation of the insulin secretory capacity and survival of pancreatic beta cells, pointing to a pivotal role for this pathway in the development of overt diabetes mellitus. PMID- 19135241 TI - Generation of porcine diploid blastocysts after injection of spermatozoa grown in nude mice. AB - It is anticipated that the utilization of spermatogonia through testicular xenografting will open new avenues for the conservation of male gametes. With the aim of establishing this new technique for genetic preservation of pigs, we used it in combination with intracytoplasmic sperm injection (ICSI). Testicular tissues derived from neonatal piglets, which contained seminiferous cords consisting of only gonocytes/spermatogonia, were transplanted under the back skin of castrated nude mice. Between 125 and 192 d after xenografting, sperm (morphologically similar to epididymal sperm) were recovered from 41 of the 65 host mice (63.1%). Testicular spermatozoa from adult boars were used as a positive control. A single spermatozoon was injected into an in vitro matured porcine oocyte, and the oocytes were electro-stimulated and cultured (graft-ICSI and testis-ICSI, respectively). Blastocyst rates in both ICSI groups (24.9% and 37.4%, respectively) were higher (P<0.05) than those without the injection procedure (parthenogenetic; 12.7%) and after injection of a small amount of injection buffer (sham; 13.0%). Rates of diploid blastocysts in both graft-ICSI and testis-ICSI groups (48.9% and 60.6%) were higher (P<0.05) than those in the parthenogenetic and sham groups (13.5% and 28.0%). Therefore, we demonstrated that porcine oocytes injected with xenogeneic sperm have in vitro developmental ability to the blastocyst stage. PMID- 19135242 TI - Failure analysis of clinically failed all-ceramic fixed partial dentures using fractal geometry. AB - OBJECTIVE: The goal of this study was to test the hypothesis that fracture toughness of the veneers in clinically failed zirconia-based fixed partial dentures (FPDs) is not significantly different from that of the in vitro group and to determine the potential reasons for their failures. METHODS: Fracture toughness values of the veneer layers in clinically failed zirconia core/glass veneer FPDs (n=4) and laboratory prepared glass veneer bar specimens (n=6) were determined using fractal analysis. A modified slit island technique was employed to measure the fractal dimensional increment (D*) of the two studied groups. The fracture toughness (K(C)) values were estimated using equation K(C)=Ea(o)(1/2)D*(1/2), where E is the elastic modulus and a(o) is a characteristic length parameter. Fracture toughness (K(C)) values of the specimens calculated using fractal analysis and fractography were statistically compared using a paired t-test. RESULTS: The average fracture toughness of the veneer in clinically FPDs (0.5+/-0.05 MPam(1/2)) is not significantly different (p>0.05) from that of the bar specimens (0.6+/-0.1 MPam(1/2)). The reasons for the early failures in FPDs could be occlusal overloading, stress corrosion, fatigue or improper structure design. SIGNIFICANCE: Fractal analysis is shown to be an alternative analytic tool for clinically failed ceramic restorations, especially for those with fracture origins chipped off during mastication and hence could not be analyzed using other techniques, such as fractography. PMID- 19135243 TI - Characterisation and modelling of marine dissolved organic matter interactions with major and trace cations. AB - A two-step protocol (nano-filtration and reverse osmosis) was applied for natural organic matter (NOM) preconcentration of a seawater sample. Complexing affinities of the so concentrated marine dissolved NOM (DNOM) towards major and trace cations were studied by potentiometric and voltammetric titration techniques. The potentiometric titration experiments fitted by models describing and characterising the DNOM-cation interactions, revealed four distinct classes of acidic sites (pKa of 3.6, 4.8, 8.6 and 12). A total acidic sites density of 445meq/mol(C) was estimated, with a majority (60%) of carboxylic-like sites. Pseudopolarographic measurements revealed two distinct groups of copper complexes: labile, reducible at about -0.2V; and inert, directly reducible at about -1.4V. Simultaneous competition between copper, calcium and proton highlighted the presence of two classes of binding sites (density of 1.72 and 10.25 meq mol(C)(-1), respectively, corresponding to 3% of total acidic sites). The first class was more specific to copper (logK(CuL) 9.9, logK(CaL) 2.5, pKa 8.6), whereas stronger competition between copper and calcium occurred for the second class (logK(CuL) 6.9, logK(CaL) 5.5, pKa 8.2). The binding sites characterisation was validated by the very good matching of the non-concentrated seawater sample titration data with the simulated curves obtained using the binding parameters from the concentrated sample. Furthermore, this comparison also validated the applied preconcentration protocol, highlighting its negligible influence on organic matter properties when considering copper complexation. PMID- 19135244 TI - Dexamethasone-containing PLGA superparamagnetic microparticles as carriers for the local treatment of arthritis. AB - Superparamagnetic iron oxide nanoparticles (SPIONs) are attractive materials that have been widely used in medicine for diagnostic imaging and therapeutic applications. In our study, SPIONs and the corticosteroid dexamethasone acetate (DXM) are co-encapsulated into PLGA microparticles for the aim of locally treating inflammatory conditions such as arthritis. The magnetic properties conferred by the SPIONs could help to maintain the microparticles in the joint with an external magnet. The aim of this study was to investigate the interaction between magnetic microparticles and human synovial fibroblasts in terms of microparticle uptake (FACS, confocal and optical microscopy), internalization mechanism (Prussian Blue staining, TEM, immunofluorescence), cell toxicity (MTT) and tissue reaction after intra-articular injection (histology). The results show that the microparticles have an excellent biocompatibility with synoviocytes and that they are internalized through a phagocytic process, as demonstrated by fluorescence-activated cell sorting and morphological analyses of cells exposed to microparticles. Histological analysis showed that the prepared microparticles did not induce any inflammatory reaction in the joint. This type of carrier could represent a suitable magnetically retainable intra-articular drug delivery system for treating joint diseases such as arthritis or osteoarthritis. PMID- 19135245 TI - Development of highly porous large PLGA microparticles for pulmonary drug delivery. AB - We report a new process of making highly-porous large polymeric microparticles for local drug delivery to the lungs by inhalation. Poly(lactic-co-glycolic acid) (PLGA) microparticles (average diameter, 10-20 microm) were made by the double emulsion method. To impart favorable aerodynamic properties, an effervescent salt ammonium bicarbonate (ABC) was included in the internal aqueous phase. ABC produced highly-porous structures in the PLGA particles as it escaped as ammonia and carbon dioxide. The fine-particle fraction (FPF) of the microparticles increased as a function of the ratio of ABC to PLGA. Microparticles prepared with 7.5%w/w (ABC/PLGA) had a mass median aerodynamic diameter (MMAD) of 4.0 +/- 1.2 microm and FPF of 32.0 +/- 9.1% when tested with Anderson Cascade Impactor (ACI) and Rotahaler. The highly-porous large particles deposited at the ACI stages corresponding to the trachea and below. The highly-porous large particles avoided phagocytosis by macrophages, while non-porous small particles were quickly taken up by the macrophages. Unlike other encapsulation methods which employ osmogens or extractable porogens, this method could encapsulate lysozyme and doxorubicin.HCl, with high encapsulation efficiency ( approximately 100% for both lysozyme and doxorubicin), in the PLGA microparticles characterized by desirable MMAD (4.5 +/- 0.6 microm lysozyme; 4.6 +/- 0.4 microm doxorubicin) and FPF (29.1 +/- 12.2% lysozyme; 33.8+/-3.6% doxorubicin). Fifty-two percent of encapsulated doxorubicin was released over 4 days from the highly-porous microparticles. This method is an efficient way of making polymeric microparticles for sustained local drug delivery by inhalation. PMID- 19135246 TI - The regulation of the gap junction of human mesenchymal stem cells through the internalization of quantum dots. AB - The delivery mechanism of CdSe/ZnS quantum dots (QDs) into cells was previously found to critically determine the biocompatibility of QDs to human adult mesenchymal stem cells, but the associated mechanism remained unknown. The present study tried to establish a link between the above phenomenon and the change in gap junction upon QD internalization. By comparing Pep-1- and PolyFect mediated QD internalizations, the connexin 43 (Cx43)-mediated gap junction intercellular communication (GJIC) of human adipose-derived adult stem cells was investigated in monolayer and in three-dimensional (3D) culture (alginate hollow spheres). The latter system offered cells more mobility, which was more similar as in vivo. The results showed that Pep-1-coated QDs, which escaped from the endo /lysosome degradation, could activate the F-actin assembly and the ERK-dependent phosphorylation of Cx43. The consequence was a reduction in Cx43-mediated GJIC. When the cells were grown in high density 3D alginate hollow spheres instead of in monolayer, the decrease of GJIC caused by the QD internalization was restored. These results indicated that the adaptability in QDs-mediated regulation of GJIC with different delivery coatings depended on the culture systems. The study also suggested that the regulation of gap junction may play a key role in QD cytotoxicity. PMID- 19135247 TI - Improved resistance to wear and fatigue fracture in high pressure crystallized vitamin E-containing ultra-high molecular weight polyethylene. AB - Higher crystallinity and extended chain morphology are induced in ultra-high molecular weight polyethylene (UHMWPE) in the hexagonal phase at temperatures and pressures above the triple point, resulting in improved mechanical properties. In this study, we report the effects of the presence of a plasticizing agent, namely vitamin E (alpha-tocopherol), in UHMWPE during high pressure crystallization. We found that this new vitamin E-blended and high pressure crystallized UHMWPE (VEHPE) has improved fatigue strength and wear resistance compared to virgin high pressure crystallized (HP) UHMWPE. This suggested different mechanisms of wear reduction and fatigue crack propagation resistance in UHMWPE. PMID- 19135248 TI - Monodisperse mesoporous superparamagnetic single-crystal magnetite nanoparticles for drug delivery. AB - In this contribution, we report a facile, gram-scale, low-cost route to prepare monodisperse superparamagnetic single-crystal magnetite NPs with mesoporous structure (MSSMN) via a very simple solvothermal method. The formation mechanism of MSSMN is also discussed and we think that Ostwald ripening probably plays an important role in this synthesis process. It is also interestingly found that the size and morphology of mesoporous Fe(3)O(4) NPs can be easily controlled by changing the amount of NaOH and 1,2-ethylenediamine (ETH). Most importantly, the MSSMN can be used as an effective drug delivery carrier. A typical anticancer drug, doxorubicin (Dox), is used for drug loading, and the release behaviors of Dox in two different pH solutions are studied. The results indicate that the MSSMN has a high drug loading capacity and favorable release property for Dox; thus, it is very promising for the application in drug delivery. PMID- 19135249 TI - The effect of surface immobilized bisphosphonates on the fixation of hydroxyapatite-coated titanium implants in ovariectomized rats. AB - Immobilized bisphosphonates (BPs) have been introduced to improve implant fixation, however, no information could be found about the efficiency of this approach in osteoporotic bone. This study was designed to evaluate the bone response to surface immobilized BPs on implants inserted in tibiae of ovariectomized (OVX) rats. Three months after bilateral ovariectomy, 40 rats were randomly assigned into four groups for implantation of hydroxyapatite-coated titanium implants with or without immobilized BPs: (1) control group (without BP treatments); (2) pamidronate (PAM) group (1mg/ml of PAM immersing); (3) ibandronate group (1mg/ml of ibandronate immersing); and (4) zoledronic acid (ZOL) group (1mg/ml of ZOL immersing). After implantation periods of 3 months, the peri-implant-bone density, trabecular microstructure, bone-implant interface and mechanical fixation of implants were evaluated by dual energy X-ray absorptiometry, micro-computed tomography, histology and push-out test. We found that three BPs triggered pronounced bone-implant integration and early bone formation around implants in OVX rats, with a rank order of ZOL>ibandronate>PAM. These results provide new evidence that immobilized BPs have positive effects on implant fixation in osteoporotic bone, in addition to their well-documented potency to inhibit implant loosening in normal bone. PMID- 19135250 TI - The reversal of hyperglycaemia in diabetic mice using PLGA scaffolds seeded with islet-like cells derived from human embryonic stem cells. AB - Islet-like cells derived from embryonic stem (ES) cells may be a promising therapeutic option for future diabetes treatment. Here, we demonstrated a five stage protocol with adding exendin-4 instead of nicotinamide finally could generate islet-like cells from human embryonic stem (ES) cells. Immunofluorescence analysis revealed a high percentage of c-peptide positive cells in the derivation. However, in addition to insulin/c-peptide, most cells also coexpressed PDX-1 (pancreas duodenum homeobox-1), glucagon, somatostatin or pancreatic polypeptide. Insulin and other pancreatic beta-cell-specific genes were all present in the differentiated cells. Insulin secretion could be detected and increased significantly by adding KCL in high glucose concentration in vitro. Furthermore, subcutaneous transplantation of scaffolds seeded with the islet-like cells or cell transplantation under kidney capsules for further differentiation in vivo could improve 6h fasted blood glucose levels and diabetic phenotypes in streptozotocin-induced diabetic SCID mice. More interestingly, blood vessels of host origin, characterized by mouse CD31 immunostaining, invaded the cell scaffold complexes. This work reveals a five-stage protocol with adding exendin-4 may be an effective protocol on the differentiation of human ES cells into islet like cells, and suggests scaffolds can serve as vehicles for islet-like cell transplantation. PMID- 19135251 TI - Influx mechanism of 2',3'-dideoxyinosine and uridine at the blood-placenta barrier. AB - The blood-placenta barrier (BPB) serves to protect the fetus from exposure to toxins, and to transport various nutrients, including nucleosides, and hormones from mother to fetus. It is known that nucleoside transporters contribute to the transfer of nucleosides and nucleoside analogues. 2',3'-Dideoxyinosine (ddI) has a nucleoside structure, and crosses the BPB. Although ddI is a substrate of several transporters, including equilibrative nucleoside transporters (ENT1 and ENT2), the transport mechanism of ddI in the placenta has not yet been characterized. Therefore, the purpose of this study was to clarify the influx mechanisms of ddI from the maternal to the fetal side, and to examine the interaction between ddI and uridine transport at the BPB. We studied ddI and uridine uptakes using a conditionally immortalized rat syncytiotrophoblast cell line, TR-TBT 18d-1, as a BPB model. The ddI uptake was temperature-dependent, Na(+)-independent and saturable. Kinetic analysis yielded K(m) values for ddI and uridine of 6.51 mM and 23.4 microM, respectively. Uridine uptake was inhibited by ENT1 and ENT2 substrates, and ddI uptake was also inhibited by substrates or inhibitors at concentrations that inhibit ENT2. Uridine uptake in Xenopus laevis oocytes expressing rat ENT2 was inhibited by 5mM ddI, in agreement with the results for TR-TBT 18d-1. Our results indicate that ddI and uridine are both taken up in part via ENT2 in TR-TBT 18d-1 cells, and therefore that ENT2 may contribute to their uptake at the BPB. PMID- 19135252 TI - Screening and cloning of multi-drug resistant genes in HL-60/MDR cells. AB - OBJECTIVE: To screen and clone multi-drug resistance (MDR) related genes in MDR acute myeloid leukemia cells (HL-60/MDR). METHODS: HL-60/MDR was established using All-Trans Retinoic Acid. With the HL-60 cells as "tester" and HL60/MDR as "driver", the cDNA library of HL-60/MDR was established by suppression subtractive hybridization. Then 12 of the resulting subtracted cDNA clones were selected for DNA sequencing and homology analysis. The obtained expressed sequence tags (ESTs) were analyzed with the GenBank BLASTN program to identify sequence homologies to known genes. RESULTS: The HL-60/MDR cells had different multi-drug resistance to six kinds of chemotherapeutic drugs. The 211 positive gene clones in differential cDNA library of HL-60/MDR cells were amplified with PCR and 46 gene clones exhibited differential expression (ratio >3). Twelve gene clones with significant differential expression (ratio >5) were screened out to homology analysis. Of these, 11 matched known genes and the rest 1 showed no significant homology to human or non-human known sequences. It was named as gene clone HA117. CONCLUSIONS: This effort provides the partial list of genes differential expressed in HL-60/MDR cells and a novel gene HA117 was found to be related to MDR. Identification of these genes contributes to our understanding of MDR development, and potentially provides candidate target genes to overcome MDR. PMID- 19135253 TI - The Swedish Environmental Classification and Information System for Pharmaceuticals--an empirical investigation of the motivations, intentions and expectations underlying its development and implementation. AB - In 2005 the Swedish Association of the Pharmaceutical Industry (LIF) initiated a national environmental classification and information system for pharmaceuticals. This investigation reports the results from a survey, conducted among the persons involved in the start-up process. The aim of this study is to generate knowledge contributing to the clarification of the motivations, expectations, and intentions underlying the development and implementation of the system. The decision to implement a classification and information system for pharmaceuticals was the result of a combination of several driving forces, mainly political pressure and a possibility to increase the industries' goodwill, while at the same time keeping the process under the industries' control. The expected possible effects of the system, other than increased goodwill, are according to this survey assumed to be low. The system offers little guidance for end-users in the substitution of one pharmaceutical for another. One possible reason for this could be that LIF needs to observe the interests of all its members' and should not affect competition. The affiliation of the involved actors correlates to how these actors view and value the system, but this has not hampered the collaborative process to develop and implement it. PMID- 19135254 TI - Effects of ranitidine and its photoderivatives in the aquatic environment. AB - This study was designed to assess the overall ecotoxicity of ranitidine, a histamine H(2)-receptor antagonist that inhibits stomach acid production. Hence, in addition to ranitidine, its main two photoderivatives, obtained by solar simulator irradiation in water, were investigated. The photoproducts were identified by their physical features. Bioassays were performed on rotifers and microcrustaceans to assess acute and chronic toxicity, while SOS Chromotest and Ames test were utilized to detect the genotoxic potential of the investigated compounds. The results showed that ranitidine did not show any acute toxicity at the highest concentration tested (100 mg/L) for all the organisms utilized in the bioassays. Chronic exposure to these compounds caused inhibition of growth population on rotifers and crustaceans. Genotoxic and mutagenic effects were especially found for one photoproduct suggesting that transformation products, as frequently demonstrated, may show effects higher than the respective parental compound. PMID- 19135255 TI - Estrogenic activity of pharmaceuticals in the aquatic environment. AB - In the last years pharmaceuticals have aroused great interest as environmental pollutants for their toxic effects towards non target organisms. This study wants to draw attention to a further adverse effect of drugs, the endocrine interference. The most representative drugs of the widespread classes in environment were investigated. The YES-test and the E-screen assay were performed to detect the capability of these substances to bind the human estrogenic receptor alpha (hER alpha) in comparison with 17beta-estradiol. Out of 14 tested pharmaceuticals, 9 were positive to YES-assay and 11 were positive to E-screen assay; in particular, Furosemide and the fibrates (Bezafibrate, Fenofibrate and Gemfibrozil) gave the maximal estrogenic response. Tamoxifen showed its dual activity as agonist and antagonist of hER alpha. PMID- 19135257 TI - Syntheses, characterization and biological activities of rare earth metal complexes with curcumin and 1,10-phenanthroline-5,6-dione. AB - Three new solid complexes have been synthesized by the reaction of rare earth(III) nitrate with the first ligand curcumin (HL) and the second ligand 1,10 phenanthroline-5,6-dione (L') in alcohol solution (pH=6.5-7.0). The composition of the complexes has been characterized by elemental analysis, molar conductivity, thermogravimetric analysis, IR, UV-vis methods. The results reveal that beta-diketone group of the first ligand to coordinates with rare earth ions in bidentate mode after deprotonated. But the second ligand uses its two N atoms coordinates with rare earth ions in bidentate mode. The general formula of the complexes is REL(3)L' (RE=Sm, Eu, Dy). The results of antibacterial activity indicated that the complexes have excellent antibacterial ability for the testing bacterium than that of curcumin. The result of agarose gel electrophoresis suggested that the complex of SmL(3)L' can cleave the plasmid DNA at physiological pH and temperature. And it was found that the cleavage process of plasmid DNA was sensitive to pH, however, adding radical scavengers almost had no effect on the DNA cleavage reaction, therefore, the cleavage of DNA by SmL(3)L' does not produce diffusible hydroxyl radicals via the Fenton reaction. PMID- 19135258 TI - New conjugates of superoxide dismutase/catalase mimetics with cyclodestrins. AB - Mimetics of antioxidant enzymes such as superoxide dismutases (SOD) or catalases are reported as potential new drugs able to reduce oxidative stress damage. In particular, manganese(III) complexes of salen-type ligands have been studied as both SOD and catalase mimetics. In this paper, we report the synthesis of two novel conjugates of salen-type ligands with the beta-cyclodextrin, the 6-deoxy-6 [(S-cysteamidopropyl(1,2-diamino)N,N'-bis(salicylidene))]-beta-cyclodextrin and the 6-deoxy-6-[(S-cysteamidopropyl(1,2-diamino)N,N'-bis(3-methoxysalicylidene))] beta-cyclodextrin, their spectroscopic characterization, and the synthesis and the characterization of their manganese(III) complexes. The SOD-like activity of the metal complexes was investigated by the indirect Fridovich method. The catalase like activity was tested using a Clark-type oxygen electrode. The peroxidase activity was tested using the ABTS (2,2'-azino-bis-(3 ethylbenzothiazoline-6-sulfonic acid)) assay. The glycoconjugation of salen manganese(III) complexes yields compounds with enhanced SOD activity. These complexes also show catalase and peroxidase activities higher than the simple salen complexes (EUK 113 and EUK 108). PMID- 19135256 TI - CD205 (DEC-205): a recognition receptor for apoptotic and necrotic self. AB - CD205 is an endocytic receptor that is expressed at high levels by cortical thymic epithelial cells and by dendritic cell (DC) subsets, including the splenic CD8+ DC population that is responsible for cross-presentation of apoptotic cell derived antigens. Antigen endocytosed via CD205 enters the MHC class I and MHC class II antigen presentation pathways and is subsequently presented to both CD4+ and CD8+ T cells. Despite the known role of CD205 in antigen uptake, the nature of the ligands bound by CD205 has not been determined, and most studies have relied on the use of monoclonal antibodies as surrogate ligands. To go beyond this approach, we created a panel of CD205-IgG fusion proteins spanning the extracellular portion of CD205 and used these to identify the physiological distribution of CD205 ligands. Our data demonstrate that two areas of the CD205 molecule, within C-type lectin-like domains (CTLDs) 3+4 and 9+10, recognise ligands expressed during apoptosis and necrosis of multiple cell types, and are additionally expressed by live cells of the dendritic cell line DC2.4. Thus, CD205 acts as a recognition receptor for dying cells, potentially providing an important pathway for the uptake of self-antigen in intrathymic and peripheral tolerance. PMID- 19135259 TI - Molecular structure and catechol oxidase activity of a new copper(I) complex with sterically crowded monodentate N-donor ligand. AB - The attempted alkylation of 1,3-bis(2'-pyridylimino)isoindoline (indH) by the use of n-BuLi and subsequent alkyl halides led to quaternization of the pyridine nitrogens and the zwitterionic monodentate N-ligand (Me(2)ind)I was formed. By the use of the ligand the copper(I) complex [Cu(I)(Me(2)ind)I(2)] was prepared and its structure determined. It was found to be good catalyst for the oxidation of 3,5-di-tert-butylcatechol (DTBCH(2)) to 3,5-di-tert-butyl-1,2-benzoquinone (DTBQ) and H(2)O(2) by dioxygen. Detailed kinetic studies revealed first-order dependence on the catalyst and dioxygen concentration and saturation type behavior with respect to the substrate. PMID- 19135260 TI - Covalent heme attachment to the protein in human heme oxygenase-1 with selenocysteine replacing the His25 proximal iron ligand. AB - To characterize heme oxygenase with a selenocysteine (SeCys) as the proximal iron ligand, we have expressed truncated human heme oxygenase-1 (hHO-1) His25Cys, in which Cys-25 is the only cysteine, in the Escherichia coli cysteine auxotroph strain BL21(DE3)cys. Selenocysteine incorporation into the protein was demonstrated by both intact protein mass measurement and mass spectrometric identification of the selenocysteine-containing tryptic peptide. One selenocysteine was incorporated into approximately 95% of the expressed protein. Formation of an adduct with Ellman's reagent (DTNB) indicated that the selenocysteine in the expressed protein was in the reduced state. The heme His25SeCys hHO-1 complex could be prepared by either (a) supplementing the overexpression medium with heme, or (b) reconstituting the purified apoprotein with heme. Under reducing conditions in the presence of imidazole, a covalent bond is formed by addition of the selenocysteine residue to one of the heme vinyl groups. No covalent bond is formed when the heme is replaced by mesoheme, in which the vinyls are replaced by ethyl groups. These results, together with our earlier demonstration that external selenolate ligands can transfer an electron to the iron [Y. Jiang, P.R. Ortiz de Montellano, Inorg. Chem. 47 (2008) 3480-3482 ], indicate that a selenyl radical is formed in the hHO-1 His25SeCys mutant that adds to a heme vinyl group. PMID- 19135261 TI - The role of traditional confinement practices in determining postpartum depression in women in Chinese cultures: a systematic review of the English language evidence. AB - BACKGROUND: The Chinese postpartum custom of "confinement" or "doing-the-month" involves formalised social support and recognition of the status of motherhood and has been presumed in anthropological literature to protect mothers of newborns from postpartum depression. The aim of this review was to examine systematically the evidence about the relationship between confinement practices and postpartum depression in Chinese cultures. METHODS: A systematic search of the English-language literature. RESULTS: Sixteen studies met inclusion criteria. It was found that the role of confinement in postpartum depression is complex: eight studies concluded that it had a protective role; four that it increased risk of postpartum mood disturbance and four studies had inconclusive findings. Aspects of the confinement practice that could contribute to or fail to protect against postpartum depression include the generally diminished social support in contemporary society, conflict with a mother-in-law and the tension experienced by modern women as they work to balance traditional with contemporary values. LIMITATIONS: Methodological differences limit meaningful comparisons between the reviewed studies and generalizations from them. CONCLUSIONS: There is little consistent evidence that confinement practices reduce postpartum depression in Chinese cultures. Specific components of confinement practices might reduce psychological distress in Chinese mothers of newborns, but these cannot be discerned from the existing evidence. Confinement cannot be presumed to be available to, welcomed by or effective for all Chinese women or to be a substitute for health service provision. PMID- 19135262 TI - Young, single and not depressed: prevalence of depressive disorder among young women in rural Pakistan. AB - BACKGROUND: The prevalence of depression is very high among adult women in Pakistan but it is not known whether such a high prevalence occurs in younger women. We aimed to assess the prevalence and correlates of depression in 16 to 18 year old unmarried women in Pakistan. METHOD: Population-based survey of all 16 to 18-year old unmarried women in one rural community in Rawalpindi District, Punjab, Pakistan. Depressive disorder and psychological distress were assessed using the Structured Clinical Interview for DSM-IV Disorders (SCID) and Self Reporting Questionnaire (SRQ) respectively. RESULTS: 337 eligible women were identified of whom 321 (95%) were interviewed. Fourteen (4.4%) had depressive disorder; one third scored 9 or more on SRQ. On multivariate analysis a high SRQ score was associated with childhood experience of poverty, father's education, stressful life events, disturbed family relationships and mother's depression. LIMITATIONS: The sample was derived from one rural community only and the results should be generalised with caution. CONCLUSION: Depressive disorder is not common in young women in rural Pakistan though distress appears common and is associated with early and recent adversity and family difficulties. These results suggest future work might aim to understand onset and prevent chronic depression. PMID- 19135263 TI - The marsupial CD8 gene locus: molecular cloning and expression analysis of the alpha and beta sequences in the gray short-tailed opossum (Monodelphis domestica) and the tammar wallaby (Macropus eugenii). AB - In eutherian mammals, CD8 is a key receptor of cytotoxic T cells and plays a pivotal role in the recognition and elimination of infected host cells by cell mediated cytotoxicity. Here, we report the molecular cloning and expression analysis of CD8alpha and CD8beta cDNAs in two marsupial species, the gray short tailed opossum and the tammar wallaby. The opossum and tammar CD8 sequences share a high degree of amino acid identity of 63% (CD8alpha) and 57% (CD8beta) to each other as well as 36-45% (CD8alpha) and 38-41% (CD8beta) with their eutherian counterparts. In addition, many of the signature features of eutherian CD8alpha and CD8beta are preserved in both marsupials including the two invariant cysteines that form the intra-chain disulphide bond in the extracellular IgSfV domain and the two hinge region cysteines involved in dimerisation between the two subunits. The p56(lck) binding motif in the cytoplasmic tail of the CD8alpha subunit is also conserved. Interestingly, the opossum CD8alpha and the tammar CD8beta sequences have a truncated cytoplasmic tail. RT-PCR analysis of CD8alpha and CD8beta transcripts in the tissues of the adult opossum and tammar showed broad tissue expression with a high level of expression observed in the lymphoid tissues of both marsupials. Furthermore, RT-PCR analysis of CD8alpha and CD8beta transcripts in the immune tissues of tammar young over the first 120 days of pouch life revealed a pattern of expression analogous to the maturation of the lymphoid tissues. This is the first report confirming the presence of CD8 in the tissues of a marsupial and will provide the tools to further analyse T cell subsets in this unique group of mammals. PMID- 19135264 TI - TRPA1 and TRPV1 activation is a novel adjuvant effect mechanism in contact hypersensitivity. AB - We have revealed that local stimulation of sensory neurons is involved in the adjuvant effect of dibutyl phthalate (DBP) in a fluorescein isothiocyanate induced mouse contact hypersensitivity model. Transient receptor potential (TRP) A1 and TRPV1 seemed to be candidate DBP targets. Here we directly demonstrated that DBP activates a subset of neurons in mouse dorsal root ganglia responsive to TRPA1 and TRPV1 agonists. TRPA1 and TRPV1 activation was further demonstrated using cultured cells expressing TRP channels. Among structurally different phthalate esters, there is a positive relationship between the activation of TRPA1- or TRPV1-expressing cells and the adjuvant effect. PMID- 19135265 TI - Fate plasticity of adult hippocampal progenitors: biological relevance and therapeutic use. AB - Adult hippocampal stem/progenitor cells (AHPs) continuously give rise to new neurons throughout life, which might be an important determinant of hippocampus dependent function. Strikingly, the fate potential of AHPs is not restricted to the neuronal lineage because AHPs can be genetically induced to generate oligodendrocytes within their in vivo niche by AHP-specific ectopic expression of the basic-helix-loop-helix (bHLH) transcription factor achaete-scute complex-like 1 (ASCL1). Fate plasticity of AHPs is controlled by cell-autonomous and also niche-dependent mechanisms. Here, we discuss the biological importance and potential therapeutic applications of retained fate plasticity of AHPs in the adult mammalian brain in addition to the future scientific inquiries indicated by this finding. PMID- 19135266 TI - The burden of Parkinson disease (PD) and concomitant comorbidities. AB - The impact of PD on health status is substantial, yet the disease burden has not been clearly defined. Using a representative sample of the community-dwelling Canadian population, we examined the burden of PD and the burden of concomitant comorbidities. Health status was compared in 261 respondents with PD from the Canadian Community Health Survey to 111,707 respondents without PD using the Health Utilities Index Mark 3 (HUI3). Differences in HUI3 scores were evaluated and selected sociodemographic covariates adjusted in ANCOVA models. The impact of specific comorbidities on the health status of PD were evaluated using ANCOVA models. Adjusted mean overall HUI3 scores were significantly lower for respondents with PD (0.61 vs. 0.83, p<0.05). The largest differences in single attribute scores were observed for ambulation (-0.11, 95% confidence interval=CI= 0.16 to -0.06), dexterity (-0.11, CI=-0.19 to -0.03), and cognition (-0.10, CI= 0.18 to -0.02). Additional burden was seen when persons with PD also had urinary incontinence (-0.22, CI=-0.40 to -0.05) or arthritis (-0.17, CI=-0.30 to -0.04). PD has detrimental effects on both physical and mental health relative to the general population. The findings provide baseline population estimates of disease burden for PD. PMID- 19135268 TI - Single coronary artery with septal course of left main coronary artery from right coronary sinus associated with coronary artery disease: multislice computed tomography (MSCT) appearance. PMID- 19135267 TI - Myocardial ischemia secondary to dual coronary artery fistulas draining into main pulmonary artery. AB - Bilateral coronary artery to pulmonary artery fistulas is an uncommon congenital anomaly. These fistulas have a clinical and embryological significance. We report a rare case of combination of right coronary and left circumflex coronary fistula draining into main pulmonary artery, who presented in the emergency room with acute pain chest. The fistula had significant 1.7:1 left to right shunt and also myocardial ischemia as demonstrated by exercise Thallium(201) SPECT myocardial imaging. He was managed conservatively during the last 5 years without any percutaneous or surgical intervention. PMID- 19135269 TI - Acute anterior myocardial infarction superimposing double left anterior descending artery: depiction at whole-heart coronary magnetic resonance angiography. AB - Double left anterior descending artery arising from the left and right coronary arteries is an extremely rare congenital coronary anomaly. We describe a patient with anterior myocardial infarction, in whom whole-heart coronary magnetic resonance angiography detected double left anterior descending artery. PMID- 19135270 TI - Is direct method of low density lipoprotein cholesterol measurement appropriate for targeting lipid lowering therapy? AB - OBJECTIVES: The aim of this study was to compare the Friedewald Formula with direct homogeneous low density lipoprotein cholesterol (LDL-C) assay for the detection of LDL-C levels. METHODS: Fasting serum samples were obtained for lipid analysis from 1001 patients. Total cholesterol (TC) and triglyceride (TG) levels were measured with enzymatic methods and the measurements of high density lipoprotein cholesterol (HDL-C) and LDL-C levels were detected using direct methods. RESULTS: The mean levels of serum TC, TG, HDL-C and LDL-C were detected with in the reference range. The LDL-C estimated by the Friedewald formula was significantly correlated (P<0.01) with the direct method but there was a negative bias among them. CONCLUSION: Laboratories cannot use direct method as a substitute for Friedewald formula because direct method has not been standardized in large populations and increase cholesterol assay costs. PMID- 19135271 TI - Microscopic polyangiitis and Kawasaki disease without overt clinical cardiovascular manifestations and with abnormal cardiovascular magnetic resonance findings. AB - Abnormal cardiovascular magnetic resonance findings were found in a patient with microscopic polyangiitis and a patient with Kawasaki disease that presented without overt clinical cardiovascular manifestations. PMID- 19135272 TI - Enhanced stent visualization: a case series demonstrating practical applications during PCI. AB - BACKGROUND: Visualization of coronary stents is increasingly challenging due to the reduction in stent strut thickness to improve deliverability. On the other hand stent expansion and precise implantation in the target vessel are important in optimizing short and long-term outcomes of stent-based revascularization. Stentboost Subtract is a novel X-ray technique that improves visualization of deployed stents in the coronary arteries. Using motion compensation and integration of multiple non-contrast projection images from a fixed gantry position, this new technique depicts stent morphology allowing assessment of stent expansion and extent of overlap with adjacent stents. We present a case series in which enhanced stent visualization (ESV) facilitated interventions. METHODS: The clinical and angiographic characteristics of 6 cases utilizing ESV were reviewed. All ESV acquisitions in this case series utilized an 8 in. field of view (FOV), 3 ml/sec. for a total of 6 ml of contrast, and placement of balloon markers in the region of interest. RESULTS: The cases presented significantly facilitated the identification of bifurcation stenting techniques, precise stent positioning, stent underexpansion and assisted with defining stent vessel wall relationship that was additive to intravascular ultrasound. CONCLUSIONS: ESV is a novel tool used in coronary interventions that facilitates non-invasive assessment of stent positioning, expansion and stent-vessel interactions. This inexpensive ESV technique is complimentary to IVUS and in some cases obviates its need. PMID- 19135273 TI - The relationship between porcine circovirus 2 antigen score and antibody titre and histology of lymph nodes in 375 euthanased sick and healthy pigs from 113 British pig farms with and without postweaning multisystemic wasting syndrome. AB - Data from a cross-sectional study of 113 British pig herds carried out in 2004 were used to investigate the associations between postweaning multisystemic wasting (PMWS) in pigs and herds and porcine circovirus 2 (PCV2) antigen score and antibody titre, and associated histological signs in lymph nodes. The sensitivity and specificity of published herd definitions for PMWS were tested on the study farms to consider the role of PCV2 in PMWS. Herds were defined as PMWS affected, -unaffected or -recovered based on current and past postweaning mortality (PWM), grower pigs with clinical signs of rapid wasting, hairiness and pallor and no other known cause of death on the farm. PCV2 antigen and antibody were not used in the definition of PMWS. In each PMWS-affected herd, up to three sick pigs with the clinical signs above and one healthy pig of a similar age were taken for postmortem examination (PME). In all other herds at least one healthy pig was taken for PME. Lymph nodes were analysed for PCV2 antigen and histological changes, and serum samples were analysed for PCV2 antibody. PCV2 antibody was present in all the herds sampled. There was a non-linear association between PCV2 antigen and antibody. There was no association between the presence of high scores of PCV2 antigen in pigs and the presence of high PWM in herds. PCV2 antigen score was significantly higher in sick than healthy pigs within farms, and high PCV2 score was associated with giant cells, coalescence and absence of germinal centres in lymph nodes. These results did not vary by PMWS affected, -unaffected or -recovered farms. PCV2 antigen was present at high scores in approximately 10% of healthy pigs on all farms. All three herd definitions of PMWS were highly sensitive, defining PMWS-affected herds as affected, but had a specificity ranging from 23% to 43%. We conclude that the current diagnostic tests for PCV2 indicated higher scores of virus in sick pigs but were not useful to define pigs or herds with PMWS. The ubiquity of PCV2 and the lack of specificity of the PCV2 tests indicate that PCV2 may be a necessary but not sufficient cause of PMWS disease. Linking this with the knowledge that the herd breakdowns occurred in a space time epidemic indicates that another infectious co-factor may be necessary for disease to occur. PMID- 19135274 TI - Rationing the public provision of healthcare in the presence of private supplements: evidence from the Italian NHS. AB - In this paper we assess the relative effectiveness of user charges and administrative waiting times as a tool for rationing public healthcare in Italy. We measure demand elasticities by estimating a simultaneous equation model of GP primary care visits, public specialist consultations and private specialist consultations, as if they were part of an incomplete system of demand. We find that for public specialist consultations, own price elasticity of demand is about -0.3, while elasticity to administrative waiting time is about -.04. No substitution exists between the demand for public and private specialists, so that user charges act as a net deterrent for over-consumption. The public provision of healthcare does not induce the wealthy to opt out. Moreover our evidence suggests that user charges and waiting lists do not serve redistributive purposes. PMID- 19135275 TI - Evaluation of patient preferences towards treatment during extended hours for patients receiving radiation therapy for the treatment of cancer: a time trade off study. AB - BACKGROUND: To reduce the waiting time between diagnosis and the start of radiation therapy, some departments have introduced appointments outside of conventional working hours, but the inconvenience this may cause to patients is unknown. We examined, from the patient's perspective, whether reduced waiting times to treatment would be sufficient to trade off against potentially inconvenient appointment times. METHOD: We interviewed patients receiving radiation therapy at a major teaching hospital between January and May 2005. Two patient groups were considered: those treated during conventional working hours (8.30 am to 4.30 pm), and those treated outside these hours. Patients were asked to trade a reduction in waiting time to the start of treatment against treatment outside conventional working hours. RESULTS: Of 129 patients interviewed, 77 were treated during conventional working hours and 52 outside these hours. Fifty-seven (44%) were male and 52 (40%) were aged over 60 years. To prefer treatment out of working hours, patients being treated during conventional working hours required a larger reduction in waiting time (odds ratio 2.36, 95% CI 0.97-5.76). Patients with curable disease and those who had made few changes in their lifestyle throughout the treatment were more likely to accept treatment outside of conventional working hours. CONCLUSION: It is impractical to satisfy the treatment-time preferences of all patients. However, many patients prefer treatment outside of normal treatment times if this would reduce the time until the start of radiation therapy. Evaluating the effect of waiting times on patients' perceptions of their disease control provides important information in allocating treatment hours and appointment times. PMID- 19135276 TI - Segmentation of short keying sequences does not spontaneously transfer to other sequences. AB - Previous research suggested that highly practiced discrete 6-key sequences are spontaneously segmented, sometimes even differently for different persons. This suggests there is some limit in the length of motor chunks that are assumed to underlie the segments in the sequence. The present experiment examined whether a segmentation pattern induced in one 6-key sequence (the prestructured sequence) determines segmentation in other 6-key sequences. The results are in line with segmentation, but showed neither transfer from the prestructured to a concurrently practiced unstructured sequence, nor to two new sequences that were carried out in a subsequent phase. Moreover, segmentation of these two new sequences was mutually different. Hence, while segmentation seems a phenomenon affecting all 6-element keying sequences, the exact segmentation pattern is not determined by that of a familiar keying sequence. Another result of the present research is that using different fingers of the same hand did slow execution rate (thus indicating effector-specific sequence learning), but the rate reduction was clearly smaller than in a previous study in which transfer to fingers of the other hand was assessed (Verwey & Wright, 2004). This is more in line with effector-specific learning being a result of sequence learning in terms of a hand based reference frame than learning to directly trigger particular effectors (i.e., the fingers). PMID- 19135277 TI - HemoglobinA1c level in healthy Thai adults: reference interval and fasting plasma glucose. AB - AIM: To establish reference interval of HbA1c IFCC in Thai. MATERIALS AND METHODS: 699 whole blood samples were used. Samples had fasting plasma glucose >or=126 mg/dl (7.00 mmol/L), renal problem or hemoglobinopathy was excluded. RESULTS: Reference interval of HbA1c IFCC was 2.90-4.90%. CONCLUSION: Effect of age should be determined. PMID- 19135278 TI - Expression analysis of nine rice heat shock protein genes under abiotic stresses and ABA treatment. AB - Expression profiles of nine rice heat shock protein genes (OsHSPs) were analyzed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The nine genes exhibited distinctive expression in different organs. Expression of nine OsHSP genes was affected differentially by abiotic stresses and abscisic acid (ABA). All nine OsHSP genes were induced strongly by heat shock treatment, whereas none of them were induced by cold. The transcripts of OsHSP80.2, OsHSP71.1 and OsHSP23.7 were increased during salt tress treatment. Expression of OsHSP80.2 and OsHSP24.1 genes were enhanced while treated with 10% PEG. Only OsHSP71.1 was induced by ABA while OsHSP24.1 was suppressed by ABA. These observations imply that the nine OsHSP genes may play different roles in plant development and abiotic stress responses. PMID- 19135279 TI - Computed tomographic angiography for low risk chest pain: seeking passage. PMID- 19135280 TI - Risk of thromboembolism varies, depending on category of immobility in outpatients. AB - STUDY OBJECTIVE: Immobility predisposes to venous thromboembolism, but this risk may vary, depending on the underlying cause of immobility. METHODS: This was a prospective, longitudinal outcome study of self-presenting emergency department (ED) patients who were from 12 hospitals and had suspected venous thromboembolism. Using explicit written criteria, clinicians recorded clinical features of each patient in the ED by using a Web-based data form. The form required one of 6 types of immobility: no immobility, general or whole-body immobility greater than 48 hours, limb (orthopedic) immobility, travel greater than 8 hours causing immobility within the previous 7 days, neurologic paralysis, or other immobility not listed above. Patients were followed for 45 days for outcome of venous thromboembolism, which required positive imaging results and clinical plan to treat. Odds ratios (ORs) were derived from logistic regression including 12 covariates. RESULTS: From 7,940 patients enrolled, 545 of 7,940 (6.9%) were diagnosed with venous thromboembolism (354 pulmonary embolism, 72 deep venous thrombosis, 119 pulmonary embolism and deep venous thrombosis). Risk of venous thromboembolism varied, depending on immobility type: limb (OR=2.24; 95% confidence interval [CI] 1.40 to 3.60), general (OR=1.76; 95% CI 1.26 to 2.44), other (OR=1.97; 95% CI 1.25 to 3.09), neurologic (OR=2.23; 95% CI 1.01 to 4.92), and travel (OR=1.19; 95% CI 0.85 to 1.67). Other significant risk factors from multivariate analysis included age greater than 50 years (OR =1.5; 95% CI 1.25 to 1.82), unilateral leg swelling (OR=2.68; 95% CI 2.13 to 3.37), previous venous thromboembolism (OR=2.99; 95% CI 2.41 to 3.71), active malignancy (OR=2.23; 95% CI 1.69 to 2.95), and recent surgery (OR=2.12; 95% CI 1.61 to 2.81). CONCLUSION: In a large cohort of symptomatic ED patients, risk of venous thromboembolism was substantially increased by presence of limb, whole-body, or neurologic immobility but not by travel greater than 8 hours. These data show the importance of clarifying the cause of immobility in risk assessment of venous thromboembolism. PMID- 19135281 TI - Randomized trial of computerized quantitative pretest probability in low-risk chest pain patients: effect on safety and resource use. AB - STUDY OBJECTIVE: We hypothesize that the presentation of a quantitative pretest probability of acute coronary syndrome would safely reduce unnecessary resource use in low-risk emergency department (ED) chest pain patients. METHODS: Randomized controlled trial of adult patients with chest pain paired with their clinicians. Patients had neither obvious evidence of acute coronary syndrome nor obvious other reason for admission. Clinicans provided their unstructured point estimate for pretest probability before randomization. Clinicans and patients in the intervention group received a printout of pretest probability of acute coronary syndrome result displayed numerically and graphically. Controls received no printout. Patients were followed for 45 days for predefined criteria of acute coronary syndrome and efficacy endpoints. Endpoints were compared between groups, with 95% confidence intervals (CIs) for differences. RESULTS: Four hundred were enrolled, and 31 were excluded for cocaine use or elopement from care. The mean pretest probability estimates of acute coronary syndrome were 4 (SD 5%) from clinicians and 4 (SD 6%) from the computer. Safety and efficacy endpoints for controls (n=185) versus intervention patients (n=184) were as follows: (1) delayed or missed diagnosis of acute coronary syndrome: 1 of 185 versus 0 of 184 (95% CI for difference -2.8% to 15.0%); (2) hospital admission with no significant cardiovascular diagnosis, 11% versus 5% (-0.2% to 11%); (3) thoracic imaging imparting greater than 5 mSv radiation with a negative result, 20% versus 9% (95% CI for difference = 3.8% to 18.0%); (4) median length of stay, 11.4 hours versus 9.2 hours (95% CI for difference = -2.9 to 7.6 hours); (5) reported feeling "very satisfied" with clinician explanation of problem on follow-up survey, 38% versus 49% (95% CI for difference = 0.9% to 21.0%); (6) readmitted within 7 days, 11% versus 4% (95% CI for difference = 2.5% to 13.2%). CONCLUSION: Presentation of a quantitative estimate of the pretest probability of acute coronary syndrome to clinicians and low-risk ED chest pain patients was associated with reduced resource use, without evidence of increased rate of premature discharge of patients with acute coronary syndrome. PMID- 19135282 TI - Invasive infection with hypermucoviscous Klebsiella pneumoniae: multiple cases presenting to a single emergency department in the United States. AB - A distinctive form of community-acquired invasive Klebsiella pneumoniae infection has been well described in Southeast Asia for more than 2 decades. The clinical syndrome includes liver abscess, bacteremia, and metastatic infection. It has recently been linked to a virulent hypermucoviscous K pneumoniae phenotype and to a specific genotype. As of 2008, there were only 2 published cases of invasive infection caused by this strain of K pneumoniae in the United States. We report 4 recent cases presenting to a single public hospital emergency department in northern California, which may signal the emergence of this clinical syndrome in North America. PMID- 19135283 TI - Evidence-based emergency medicine/rational clinical examination abstract: The evidence-based diagnosis of deep venous thrombosis. PMID- 19135284 TI - Metastatic malignant pleural mesothelioma of the tongue: report of a case. PMID- 19135285 TI - Assessment and management of high-risk pregnancies in women with thrombophilia. AB - Pregnancy is an acquired hypercoagulable state which can lead to gestational vascular complications especially in the presence of other prothrombotic risk factors. The use of anticoagulation for prevention of pregnancy complications in women with thrombophilia is becoming more frequent. Efficacy and safety issues of therapeutic and prophylactic regimens use and monitoring of LMWH therapy in pregnancy are discussed in this review. In addition, a scoring system for women with thrombophilia is proposed which includes severity of pregnancy outcomes, thrombotic history and type of thrombophilia. Validation of this scoring system has revealed a significant correlation between the proposed score and LMWH dosages that were prescribed to the studied pregnant women. Careful diagnosis, observation and monitoring can add significant benefit to LMWH therapy during pregnancy. PMID- 19135286 TI - Spatial and environmental connectivity analysis in a cholera vaccine trial. AB - This paper develops theory and methods for vaccine trials that utilize spatial and environmental information. Satellite imagery is used to identify whether households are connected to one another via water bodies in a study area in rural Bangladesh. Then relationships between neighborhood-level cholera vaccine coverage and placebo incidence and neighborhood-level spatial variables are measured. The study hypothesis is that unvaccinated people who are environmentally connected to people who have been vaccinated will be at lower risk compared to unvaccinated people who are environmentally connected to people who have not been vaccinated. We use four datasets including: a cholera vaccine trial database, a longitudinal demographic database of the rural population from which the vaccine trial participants were selected, a household-level geographic information system (GIS) database of the same study area, and high resolution Quickbird satellite imagery. An environmental connectivity metric was constructed by integrating the satellite imagery with the vaccine and demographic databases linked with GIS. The results show that there is a relationship between neighborhood rates of cholera vaccination and placebo incidence. Thus, people are indirectly protected when more people in their environmentally connected neighborhood are vaccinated. This result is similar to our previous work that used a simpler Euclidean distance neighborhood to measure neighborhood vaccine coverage [Ali, M., Emch, M., von Seidlein, L., Yunus, M., Sack, D. A., Holmgren, J., et al. (2005). Herd immunity conferred by killed oral cholera vaccines in Bangladesh. Lancet, 366(9479), 44-49]. Our new method of measuring environmental connectivity is more precise since it takes into account the transmission mode of cholera and therefore this study validates our assertion that the oral cholera vaccine provides indirect protection in addition to direct protection. PMID- 19135287 TI - The link between local environment and obesity: a multilevel analysis in the Lisbon Metropolitan Area, Portugal. AB - Although individual factors have been shown to predict weight gain, contextual determinants have also attracted attention, with some authors stressing the role played by deprivation, urban sprawl, social capital and safety. Recent evidence has implicated environmental factors that facilitate the consumption of excess calories and/or make it more difficult to expend them in routine physical activity. The interrelationships found in some places between physical and social environments (key mediators) and body mass index (BMI), as well as the potential that exists for the development of healthier places, mean that more research is required into the contextual determinants of health. In Portugal, particularly in the Lisbon Metropolitan Area (LMA), the effects of physical and social environments on physical activity and BMI have not previously been explored in any detail. This study aims to highlight the associations between residential (physical and social) environment and the risk of weight gain and obesity, over and above individual attributes, assessing which indicators are the best predictors of excess weight in the LMA. The study involved data from 7669 individuals aged 18 and over from 143 neighbourhoods. Self-reported body height and weight were used to define overweight body mass index (BMI> or =25). BMI and individual (socio-demographic and behavioural) characteristics were linked to contextual data and analysed in a multilevel framework. Our findings show that different environmental factors are significantly associated with excess weight and obesity, either directly or indirectly (e.g. health-related behaviours such as eating patterns and physical activity, which are key mediators), after adjustment for individual characteristics. The results suggest that a deeper understanding of these mechanisms is critical if we want to tackle the obesity epidemic, and that policies aimed at weight control and obesity reduction must address people and places in order to bear fruit. PMID- 19135288 TI - Chronic catastrophes: exploring the concentration and sustained nature of ambulatory prescription drug expenditures in the population of British Columbia, Canada. AB - Previous research has shown that a small proportion of the population accounts for a substantial proportion of spending on physician and hospital services. Much less is known about the high-cost users of ambulatory prescription medicines. We investigate the concentration and sustained nature of ambulatory prescription drug expenditures among residents of British Columbia, Canada in 2001 and 2004. Linking person-specific administrative data from several sources, we examine the demographics, socio-economic status, and health status of high-cost ambulatory pharmaceutical users and the extent that high-cost pharmaceutical use was sustained, at the individual level, from 2001 to 2004. The top 5% of users were responsible for 48% of ambulatory prescription expenditures in the province. A significant burden of morbidity, as well as sustained high expenditures, characterized these users. They were older, more likely to be female, more likely to be of low income, and more likely to be hospitalized and die within the year of study than other pharmaceutical users and non-users. Our results suggest that careful consideration should be given to the long-term financial burdens and access barriers created by pharmaceutical insurance policies that rely heavily on private payments by individuals. Our focus is on costs associated with ambulatory prescription drug use, however, had we included information on the cost of prescription drugs used in hospitals, we would likely have detected an even stronger relationship between high-cost pharmaceutical use and poor health status. PMID- 19135289 TI - In vitro demineralization of human enamel natural and abraded surfaces: a micromechanical and SEM investigation. AB - OBJECTIVES: To compare the demineralization pattern that occurs in abraded samples and unabraded ones quantitatively and microscopically using nanoindentation and SEM. METHODS: Using 12 human third molars, one half of each tooth was abraded to a depth approximately 200 microm below the outer surface enamel, and the other half was left untreated. All specimens were demineralized for 3d, 5d, 7d and 14d respectively. The cross sections of all lesions were evaluated with nanoindentation and SEM. RESULTS: Unabraded samples had least loss of mechanical properties and lesion depth compared to abraded samples, with most mechanical property loss occurring within a shallower layer of the lesion. The variation of mechanical properties in unabraded samples was wider than abraded samples. SEM images showed a characteristic "keyhole" structure for both samples after demineralization, with the rod core extensively demineralized while the interrod remained intact. Acid attack initiated at the rod sheath space then penetrated into the rod core before extending into neighbouring rods through the rod tail. CONCLUSION: Abraded samples exposed to in vitro demineralization form deeper lesion depths and greater loss of mechanical properties than unabraded samples subjected to the same demineralization. Unabraded samples manifested characteristic subsurface demineralization with a shallow surface layer that remained intact, whereas no intact surface was found in abraded samples. The demineralization pattern of unabraded samples more closely resembled the pattern of natural white spot lesions and displayed wide inter-sample variation. Consideration should be given to experimental design with unabraded teeth specimens for future demineralization studies. PMID- 19135290 TI - Maternal, gestational and neonatal characteristics and maternal angiogenic factors in normotensive pregnancies. AB - OBJECTIVE: Alterations in maternal circulating angiogenic factors are proposed to result in hypertension and proteinuria and development of preeclampsia. The aim of this study was to explore whether preeclampsia risk factors are associated with maternal angiogenic profile in normotensive pregnancies. STUDY DESIGN: Associations of pregnancy characteristics and maternal serum concentrations at delivery of proangiogenic placental growth factor (PlGF), antiangiogenic soluble fms-like tyrosine kinase receptor (sFlt1) and soluble endoglin (sEng), as well as the antiangiogenic ratios sFlt1/PlGF and (sFlt1+sEng)/PlGF were analyzed in 43 normotensive and 44 preeclamptic pregnancies. RESULTS: In normotensive pregnancies, increasing maternal age was associated with a more antiangiogenic profile, including lower PlGF concentrations and a higher (sFlt1+sEng)/PlGF ratio (P<0.05). In preeclampsia, shorter length of gestation and lower birth weight percentile were associated with a more antiangiogenic profile. CONCLUSION: A greater antiangiogenic profile with older maternal age may suggest a biological mechanism which mediates this preeclampsia risk factor. In preeclampsia, the antiangiogenic state was more pronounced with clinical characteristics indicative of greater disease severity. PMID- 19135291 TI - Metoclopramide and diphenhydramine in the treatment of hyperemesis gravidarum: effectiveness and predictors of rehospitalisation. AB - OBJECTIVES: Hyperemesis gravidarum (HG) is the second most common reason for hospitalisation during pregnancy. Since 2002, a new HG treatment protocol consisting of metoclopramide plus diphenhydramine was put in place at CHU Sainte Justine, affiliated to University of Montreal, Quebec, Canada. The objectives of this study were to evaluate the effectiveness of this new HG protocol regarding length of hospitalisation for HG, rate of rehospitalisation, evolution of nausea and vomiting symptoms, and rate of adverse events. STUDY DESIGN: A retrospective cohort study was conducted from 2002 to 2006 on the population of pregnant women diagnosed with HG, and treated at CHU Sainte-Justine with the new protocol consisting of intravenous metoclopramide 1.2-1.8 mg/h plus diphenhydramine 50 mg every 6 h. These women were compared to a historical control group consisting of women diagnosed with HG, and treated in the same institution with intravenous droperidol 0.5-1 mg/h plus diphenhydramine 25-50 mg every 6h between 1998 and 2001. RESULTS: During the study period, a total of 130 pregnant women were exposed to the new HG protocol versus 99 that were exposed to the droperidol and diphenhydramine combination between 1998 and 2001. Our study showed that the new HG protocol was associated with a greater improvement of vomiting symptoms (36% vs. 21%; p=0.0397), and with fewer adverse events. The new HG protocol was equivalent to the droperidol and diphenhydramine combination to reduce nausea symptoms, length of hospitalisation (3.7 days vs. 3.1 days), and rate of rehospitalisation for HG (19.23% vs. 24.44%). CONCLUSIONS: The new protocol consisting of the combination of metoclopramide and diphenhydramine appears to be a good option in the management of hyperemesis gravidarum. PMID- 19135292 TI - Comparative assessment of antimicrobial activities of antibiotic-treated penile prostheses. AB - BACKGROUND: Although infections associated with penile implants are relatively infrequent, they result in serious medical consequences. Because treatment of these infections usually requires removal of the infected penile implant, prevention of infection is crucial. Since bacterial colonization of the implant is a prelude to clinical infection, antimicrobial modification of the devices may inhibit device colonization and subsequent infection. OBJECTIVE: We compared the spectrum and durability, both in vitro and in vivo, of two antibiotic-treated penile prostheses: InhibiZone implants pre-impregnated with minocycline and rifampin (M/R) and Titan implants dipped in vancomycin. DESIGN, SETTING, AND PARTICIPANTS: 1*1-cm cylinder segments of (1) control untreated, (2) M/R impregnated, and (3) vancomycin-dipped implants were studied. Baseline zones of inhibition (ZI) were determined against clinical isolates, including methicillin resistant Staphylococcus aureus (MRSA), methicillin-resistant S. epidermidis (MRSE), vancomycin-resistant Enterococcus (VRE), and Escherichia coli. In addition, ZI against methicillin-susceptible S. aureus were compared both in vitro after being washed in a flow chamber and after subcutaneous implantation in rabbits for 1, 2, 7, and 14 d. MEASUREMENTS: ZI were measured as the diameter of the clear zone around each test device minus the external diameter of the device. RESULTS AND LIMITATIONS: Implants pre-impregnated with M/R displayed a broader spectrum of antimicrobial activity than vancomycin-dipped implants against both gram-positive and -negative bacteria. The M/R-impregnated devices also yielded significantly larger zones of inhibition against S. aureus than vancomycin-dipped implants, both in vitro (p<0.003) and in vivo throughout the 14-d period of device implantation in rabbits (p<=0.03). CONCLUSIONS: Penile prostheses impregnated with M/R have a broader spectrum in vitro and a more durable antimicrobial activity in vitro and in an animal model than implants dipped in vancomycin. Therefore, along with being a more practical model for incorporating antimicrobials onto the device, the use of implants pre-impregnated with M/R may help reduce the incidence of penile implant infection. PMID- 19135293 TI - Editorial comment on: Comparative assessment of antimicrobial activities of antibiotic-treated penile prostheses. PMID- 19135294 TI - Editorial comment on: Comparative assessment of antimicrobial activities of antibiotic-treated penile prostheses. PMID- 19135295 TI - Population dynamic succession and quantification of ammonia-oxidizing bacteria in a membrane bioreactor treating municipal wastewater. AB - Ammonia-oxidizing bacterial community structures and abundances in membrane bioreactor (MBR), which was utilized to treat municipal wastewater, were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), cloning of 16S rDNA fragments and quantitative real-time PCR techniques. DGGE analysis indicated marked change of ammonia-oxidizing bacterial communities' structure mainly happened during day 62 to day 80. The results suggest that the dynamics of AOB community structure are important and necessary for the improving and stable nitrogen degradation and the removal in MBR. Cloning and sequencing analyses of screened clones revealed multitude nitrosifying organisms and denitrifying bacteria, which suggest that they may coexist several modes of nitrification and denitrification in MBR. Real-time PCR results demonstrated that the abundance of AOB was <0.01% of the total bacteria in inoculating activated sludge. After domestication period, AOB 16S rDNA and amoA gene content in 120th day was 20.44 and 14.38 times those of seeding activated sludge. The analyses suggest a correlation between the quantity of AOB populations and nitrification efficiency. PMID- 19135296 TI - Studies on the interaction of olive cake and its hydrophylic extracts with polyvalent metal ions (Cu(II), Eu(III)) in aqueous solutions. AB - The paper presents and discusses the chemical affinity of two different forms of natural organic matter (e.g. in solid (SOM) and soluble form (DOM)) for Cu(2+) and Eu(3+) ions. In the this study SOM is represented by olive cake and DOM by its hydrophilic extracts. The investigations were performed in aqueous 0.1M NaClO(4) solutions at pH 6, 23 degrees C and under atmospheric conditions by means of potentiometry using a copper ion selective electrode and competitions reactions between the two metal ions. The experimental data were evaluated according to an "operational" approach, which is based on the charge neutralization model and the Scatchard approach. Evaluation of the data resulted in the determination of conditional formation constants (beta(*)), which are a measure for the chemical affinity of natural organic matter for a metal ion. For the olive cake metal ion binding the values of log beta(*) for Cu(II) and Eu(III) were evaluated to be 5.1+/-0.3 and 5.4+/-0.5, respectively, whereas for the hydrophilic extracts the corresponding values for the Cu(II) and Eu(III) complexes were evaluated to be 5.3+/-0.3 and 6.3+/-0.5, respectively. The beta(*) values show that natural organic matter in the two different forms presents almost similar affinity for the Cu(2+) ion, whereas the affinity of DOM for the Eu(3+) ion is significantly higher than the corresponding affinity of SOM. This is ascribed to the higher flexibility of DOM, which allows better coordination of its active sites around the Eu(3+) ion. PMID- 19135297 TI - Ability of the Fire Propagation Apparatus to characterise the heat release rate of energetic materials. AB - Energetic materials encompass a wide range of chemical compounds. They react very rapidly releasing large amounts of energy. One of their peculiarities is that they carry an oxidizer and do not require oxygen from the air as their primary reaction partner. The aim of this paper is to present an analysis of the ability to estimate the heat release rate of a sample energetic material using two calorimetric methodologies. The methods are based on Oxygen Consumption and Carbon Dioxide Generation principles. Data have been obtained from experiments carried out with the Fire Propagation Apparatus. First, results from smoke powder combustion tests reveal significant discrepancies between the two approaches. Results from a sensitivity analysis realised in a previous work underlined that the most likely parameters to alter the heat release rate estimation are the energy constants and the concentration of oxygen. Correction procedures have been developed; one based on the estimation of the amount of oxygen supplied by the oxidizer, and a second one based on the calculation of new energy constants accounting for the chemical decomposition of the tested materials. Results are presented in this study. PMID- 19135298 TI - Enhanced mercury ion adsorption by amine-modified activated carbon. AB - Mercury (Hg) is one of the most toxic metals found in water and sediments. In an effort to develop an effective adsorbent for aqueous Hg removal, activated carbon (AC) was modified with an amino-terminated organosilicon (3 aminopropyltriethoxysilane, APTES). Surface properties of the APTES-modified AC (MAC) were characterized by the scanning electron microscopy in conjunction with the energy-dispersive spectroscopy (SEM-EDS), the Fourier transform infrared spectroscopy (FT-IR), and potentiometry. The impacts of solvent, APTES concentration, reactive time and temperature on the surface modification were evaluated. The aqueous Hg adsorptive kinetics and capacity were also determined. Results demonstrated that the strong Hg-binding amine ligands were effectively introduced onto the AC surfaces through the silanol reaction between carbon surface functional groups (-COOH, -COH) and APTES molecules. The modification lowered the pH at the point of zero charge (pH(pzc)) to 4.54 from 9.6, favoring cation adsorption. MAC presented a faster rate of the Hg (II) adsorption and more than double adsorptive capacity as compared with AC. PMID- 19135299 TI - Synthesis, characterization and performance in arsenic removal of iron-doped activated carbons prepared by impregnation with Fe(III) and Fe(II). AB - Arsenic removal from natural well water from the state of Chihuahua (Mexico) is investigated by adsorption using a commercial activated carbon (AC). The latter is used as such, or after oxidation by several chemicals in aqueous solution: nitric acid, hydrogen peroxide, and ammonium persulphate. Raw and oxidised activated carbons are fully characterised (elementary analysis, surface chemistry, pore texture parameters, pH(ZC), and TEM observation). Adsorption of As is measured in the aforementioned water, containing ca. 300 ppb of arsenic: removal of As is poor with the raw AC, and only the most oxidised carbons exhibit higher performances. By contrast, iron-doped ACs are much more efficient for that purpose, though their As uptake strongly depends on their preparation conditions: a number of samples were synthesised by impregnation of raw and oxidised ACs with HCl aqueous solutions of either FeCl(3) or FeCl(2) at various concentrations and various pH. It is shown that iron(II) chloride is better for obtaining high iron contents in the resultant ACs (up to 8.34 wt.%), leading to high As uptake, close to 0.036 mg As/g C. In these conditions, 100% of the As initially present in the natural well water is removed, as soon as the Fe content of the adsorbent is higher than 2 wt.%. PMID- 19135300 TI - Improving the ammonium ion uptake onto natural zeolite by using an integrated modification process. AB - Detailed investigation on the development of physical and chemical properties of a natural calcium-rich zeolite modified by an integrated process, as well as the relation between the development and ammonium ion uptake (AIU), was conducted. This process consisted of pretreatment (grinding and sieving), sodium salt modification and calcination. Both pretreatment and salt modification largely increased BET surface area, total pore volume and average pore diameter of the raw zeolite. Individual calcination at temperature above 150 degrees C caused framework collapse, losses of partial clinoptilolite and production of X-ray amorphous material, resulting in pore blockage and the decreases in pore volume and BET surface area. However, the introduction of sodium ion enhanced the heat resistance of the raw material from 150 to 400 degrees C, and Na(+) ion treatment followed by calcination could effectively improved pore and surface properties of zeolite, thus leading to the significant enhancement in ammonium ion exchange and adsorption capabilities. PMID- 19135301 TI - Comparative sorption of benzo[alpha]phrene to different humic acids and humin in sediments. AB - Sediment/soil organic matter (SOM) is the predominant sorbents for hydrophobic organic compounds (HOCs). The identification and comparison of sorption characterization of HOCs by different organic matter fractions are essential to predict the fate and transport of HOCs in soils and sediments. The objectives of this paper are to compare the sorption of benzo[alpha]phrene (BaP) to the humic acid (HA) and humin (HM) extracted from different sediments. The HA and HM were extracted with 0.1M NaOH from five sediments in different areas in China, and their sorption isotherms for BaP were determined. All sorption isotherms were nonlinear and nonlinearity increased in the order HA98%, which denote on satisfactorily application of the method. PMID- 19135304 TI - Simultaneous spectrophotometric determination of binary mixtures of surfactants using continuous wavelet transformation. AB - This work presents a simple, rapid, and novel method for simultaneous determination of binary mixtures of some surfactants using continuous wavelet transformation. The method is based on the difference in the effect of surfactants Cetyltrimethylammoniumbromide (CTAB), dodecyl trimethylammonium bromide (DTAB), cetylpyridinium bromide (CPB) and TritonX-100 (TX-100) on the absorption spectra of complex of Beryllium with Chrome Azurol S (CAS) at pH 5.4. Binary mixtures of CTAB-DTAB, DTAB-CPB and CTAB-TX-100 were analyzed without prior separation steps. Different mother wavelets from the family of continuous wavelet transforms were selected and applied under the optimal conditions for simultaneous determinations. The proposed methods, under the working conditions, were successfully applied to simultaneous determination of surfactants in hair conditioner and mouthwash samples. PMID- 19135305 TI - Selective removal of Cr(VI) ions from aqueous solutions including Cr(VI), Cu(II) and Cd(II) ions by 4-vinyl pyridine/2-hydroxyethylmethacrylate monomer mixture grafted poly(ethylene terephthalate) fiber. AB - In this study, a new reactively fibrous adsorbent was prepared by grafting 4 vinyl pyridine (4-VP) and 2-hydroxyethylmethacrylate (HEMA) monomer mixture onto poly(ethylene terephthalate) (PET) fibers for removal of Cr(VI), Cu(II) and Cd(II) metal ions from aqueous solution by using batch adsorption method. The influence of various parameters such as graft yield (GY), pH, adsorption time, initial ion concentration and adsorption temperature was investigated. The selectivity of the reactive fiber was also examined. The results show that the adsorbed amount of metal ions followed as given in the order Cr(VI)>Cd(II)>Cu(II). At pH 3, Cr(VI) was removed by 99% while the initial concentration of ions was at 5 mg L(-1) and by 94% at 400 mg L(-1). It was found that the grafted fiber is more selective for Cr(VI) ions in the mixed solution of Cr(VI)-Cu(II), Cr(VI)-Cd(II) and Cr(VI)-Cu(II)-Cd(II) at pH 3 and it was observed that the grafted fibers are stable and regenerable by acid and base without losing their activity. PMID- 19135306 TI - Agricultural soil monitoring of PCDD/Fs in the vicinity of a municipal solid waste incinerator in Eastern China: temporal variations and possible sources. AB - The temporal variations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in 33 agricultural soil samples in the vicinity of a municipal solid waste incinerator (MSWI) in Eastern China were determined one year after the initial investigation in 2006. The soil PCDD/F concentrations in 2007 ranged from 73.6 to 377 ng kg(-1) (0.60-6.38 ng I-TEQ kg(-1)). During 2006-2007, the overall soil PCDD/F levels increased significantly, i.e., 33% and 39% for total concentration and I-TEQ (median value), respectively. Moreover, soils in the study area are proved to be almost free from previously suspected PCDD/F sources, i.e., pentachlorophenol/sodium penta-chlorophenate (PCP/PCP-Na) and 1,3,5 trichloro-2-(4-nitrophenoxy) benzene (CNP) contaminations. Furthermore, the results from a congener-specific factor analysis between soils (collected in two investigations) and dioxin emission sources suggest that diffuse sources including open burning of wastes, traffic and hot water boilers are major contributors that are responsible for the accumulation of PCDD/Fs in soils. By contrast, the impact of the presumably major PCDD/F source identified in our previous study, i.e., the MSWI, seems to be limited. PMID- 19135307 TI - A new induction period based reaction rate method for determination trace amounts of phenylhydrazine in water samples. AB - A simple, selective, and sensitive kinetic spectrophotometric method is described for the determination of trace amounts of phenylhydrazine, which is based on its inhibition effect on the reaction between meta cresol purple (MCP) and periodate in the presence of bromide ions. The reaction was monitored spectrophotometrically by measuring the change in absorbance of MCP at 525 nm. The calibration graph was linear in the range of 1.0-10.0 microM. The detection limit (3 sigma) was 0.020 microM. The relative standard deviations for 10 replicate measurements of 3.0, 5.0 and 7.0 microM of phenylhydrazine were 2.0%, 1.4%, and 0.90%, respectively. The potential interfering substances were studied in the presence of phenylhydrazine. The proposed method was applied to the analysis of water samples. PMID- 19135308 TI - Surface segregation of Au-Pd alloys in UHV and reactive environments: quantification by a catalytic atom probe. AB - The surface composition of an Au-62at%Pd alloy has been studied by means of a catalytic atom probe (CAP) before and after exposures to nitric oxide (NO) at temperatures ranging from 300 to 573K for 20min. Subsequent CAP analysis at 100K revealed a considerable surface enrichment in Pd (to approximately 80at%) after exposure at 573K. This is correlated with the occurrence of NO dissociation, and the formation of strong Pd-O bonds at the surface. Blank experiments in ultra high vacuum reflect the surface composition of the bulk material, in excellent agreement with electron microprobe analysis. At 573K, no detectable surface segregation occurs in the absence of NO adsorption for the times and temperatures studied. However, classical Metropolis Monte-Carlo simulations performed with a semi-empirical potential on the Au(40)Pd(60) (111), (110) and (100) systems show surface enrichment of gold at equilibrium. This suggests that the temperatures of the clean surface segregation experiments are too low to reach equilibrium within times of the order of hours. PMID- 19135309 TI - Anaplasma marginale infection in a Japanese Black cow 13 years after eradication of Rhipicephalus (Boophilus) microplus in Okinawa, Japan. AB - In October 2007, a 15-year-old Japanese Black cow on Ishigaki Island, Okinawa, Japan, was diagnosed with Anaplasma marginale infection based on clinical symptoms, blood examination, smear observation, 16S rRNA and groEL gene sequence analysis, and the result of a CF test. The cow was introduced into the farm from mainland Japan as a calf in 1993, one year before the eradication of Rhipicephalus (Boophilus) microplus, the main vector of A. marginale in Okinawa Prefecture. It is possible that the cow was first infected with A. marginale as a calf in Ishigaki Island and had been persistently infected since then. This is the first reported clinical case of A. marginale infection of cattle since the eradication of R. microplus in Okinawa Prefecture. Additional analysis of major surface protein 1alpha amino acid sequences revealed that the A. marginale Okinawa strain presented four new repeat forms which were not seen in other strains. This indicates that the Okinawa strain may be a unique geographical variant of A. marginale. PMID- 19135310 TI - Dose confirmation studies for monepantel, an amino-acetonitrile derivative, against fourth stage gastro-intestinal nematode larvae infecting sheep. AB - Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Nine dose confirmation studies were conducted in Australia, New Zealand and Switzerland to confirm the minimum therapeutic oral dose of monepantel to control fourth stage (L4) gastro-intestinal nematode larvae in sheep (target species were Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirusbattus, Nematodirusfilicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum). In each study, sheep infected with a defined selection of the target nematodes were treated with 2.5mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. The results demonstrate high (95<100%) efficacy of monepantel when administered orally to sheep at 2.5mg/kg for most species tested. Efficacy levels against N. spathiger and O. venulosum were variable and failed to meet the required regulatory standard (> or =90%) in some studies. Efficacy was demonstrated against L4 stages of nematodes known to be resistant to either benzimidazole and/or levamisole anthelmintics (macrocyclic lactone resistant isolates were not available for testing). The broad-spectrum activity of monepantel against L4 larvae of common gastro-intestinal nematodes in sheep and its favorable safety profile represents a significant advance in the treatment of parasitic gastro-enteritis in this animal species. No adverse effects related to treatment with monepantel were observed. PMID- 19135311 TI - A new minimum legal drinking age (MLDA)? Some findings to inform the debate. AB - Despite support that the minimum legal drinking age (MLDA) of 21 is an important public health provision in the United States, a group of college presidents are petitioning for the MLDA to be lowered. To inform this debate, we examined whether heavy and risky drinking was associated with a stance to lower the MLDA in 865 college students under the age of 21. Results showed that, in comparison to other students, heavy/risky drinkers more often had a stance to lower the MLDA. Thus, for students, the MLDA debate seems to be less a philosophical issue about prohibition and harm reduction, and appears to be more a political stance that reflects students' concurrent behaviors. We discuss how drinking and MLDA laws in Europe might compare with the United States, and how future policy work might benefit from empirical and cross-cultural study. PMID- 19135313 TI - The value of the internship for radiation oncology training: results of a survey of current and recent trainees. AB - PURPOSE: Although a 12-month clinical internship is the traditional precursor to a radiation oncology residency, the continuance of this mandated training sequence has been questioned. This study was performed to evaluate the perceptions of current radiation oncology residents with respect to the value of their internship experience. METHODS AND MATERIALS: A survey was sent to all US radiation oncology residents. Each was queried about whether they considered the internship to be a necessary prerequisite for a career as a radiation oncologist and as a physician. Preferences were listed on a Likert scale (1 = not at all necessary to 5 = absolutely necessary). RESULTS: Seventy-one percent considered the internship year mostly (Likert Scale 4) or absolutely necessary (Likert Scale 5) for their development as a radiation oncologist, whereas 19.1% answered hardly or not at all (Likert Scale 2 and 1, respectively). With respect to their collective considerations about the impact of the internship year on their development as a physician, 89% had a positive response, 5.8% had a negative response, and 4.7% had no opinion. Although both deemed the preliminary year favorably, affirmative answers were more frequent among erstwhile internal medicine interns than former transitional program interns. CONCLUSIONS: A majority of radiation oncology residents positively acknowledged their internship for their development as a specialist and an even greater majority valued it for their development as a physician. This affirmative opinion was registered more frequently by those completing an internal medicine internship compared with a transitional internship. PMID- 19135312 TI - The pattern of failure after reirradiation of recurrent squamous cell head and neck cancer: implications for defining the targets. AB - PURPOSE: Reirradiation (re-RT) of recurrent head and neck cancer (HNC) may achieve long-term disease control in some patients, at the expense of high rates of late sequelae. Limiting the re-RT targets to the recurrent gross tumor volume (rGTV) would reduce the volumes of reirradiated tissues; however, its effect on tumor recurrence pattern is unknown. METHODS AND MATERIALS: This is a retrospective review of 66 patients who underwent curative-intent re-RT for nonresectable recurrent or second primary mucosal squamous cell HNC. Treatment was delivered with three-dimensional conformal (3D) RT or intensity-modulated RT (IMRT). The targets in all patients consisted of the rGTVs with tight (0.5-cm) margins, with no intent to treat prophylactically lymph nodes or subclinical disease in the vicinity of the rGTVs. The sites of locoregional failures (LRFs) were determined using imaging at the time of failure and were compared with the rGTVs. RESULTS: Median re-RT dose was 68 Gy. Forty-seven patients (71%) received concomitant chemotherapy, and 31 (47%) received hyperfractionated, accelerated RT. At a median follow-up of 42 months, 16 (23%) were alive and disease-free. Fifty patients (77%) had a third recurrence or persistent disease, including 47 LRFs. All LRFs occurred within the rGTVs except for two (4%) (95% confidence interval, 0-11%). Nineteen patients (29%) had Grade > or = 3 late complications, mostly dysphagia (12 patients). CONCLUSIONS: Almost all LRFs occurred within the reirradiated rGTVs despite avoiding prophylactic RT of tissue at risk of subclinical disease. These results support confining the re-RT targets to the rGTVs to reduce reirradiated tissue volumes. PMID- 19135314 TI - Neoplastic transformation induced by carbon ions. AB - PURPOSE: The objective of this experiment was to compare the oncogenic potential of carbon ion beams and conventional photon beams for use in radiotherapy. METHODS AND MATERIALS: The HeLa X human skin fibroblast cell line CGL1 was irradiated with carbon ions of three different energies (270, 100, and 11.4 MeV/u). Inactivation and transformation data were compared with those for 15 MeV photons. RESULTS: Inactivation and transformation frequencies for the 270 MeV/u carbon ions were similar to those for 15-MeV photons. The maximal relative biologic effectiveness (RBE(alpha)) values for 100MeV/u and 11.4 MeV/u carbon ions, respectively, were as follows: inactivation, 1.6 +/- 0.2 and 6.7 +/- 0.7; and transformation per surviving cell, 2.5 +/- 0.6 and 12 +/- 3. The curve for dose-transformation per cell at risk exhibited a maximum that was shifted toward lower doses at lower energies. CONCLUSIONS: Transformation induction per cell at risk for carbon ions in the entrance channel was comparable to that for photons, whereas for the lower energies, 100 MeV/u and 11 MeV/u, which are representative of the energies delivered to the tumor margins and volume, respectively, the probability of transformation in a single cell was greater than it was for photons. In addition, at isoeffective doses with respect to cell killing, the 11.4-MeV/u beam was more oncogenic than were photons. PMID- 19135315 TI - Risk factors for hearing loss in patients treated with intensity-modulated radiotherapy for head-and-neck tumors. AB - PURPOSE: Radiotherapy (RT) is a common treatment of head-and-neck carcinoma. The objective of this study was to perform a prospective multivariate assessment of the dose-effect relationship between intensity-modulated RT and hearing loss. METHODS AND MATERIALS: Pure tone audiometry at 0.250-16 kHz was obtained before and after treatment in 101 patients (202 ears). All patients received full-course intensity-modulated RT (range, 56-70 Gy), with a median cochlear dose of 11.4 Gy (range, 0.2-69.7). RESULTS: Audiometry was performed 1 week before and a median of 9 weeks (range, 1-112) after treatment. The mean hearing deterioration at pure tone average air-conduction 1-2-4 kHz was small (from 28.6 dB HL to 30.1 dB HL). However, individual patients showed clinically significant hearing loss, with 10 dB threshold shift incidences of 13% and 18% at pure tone averages air-conduction 1-2-4 kHz and 8-10-12.5 kHz, respectively. Post-treatment hearing capability was unfavorable in the case of greater inner ear radiation doses (p <0.0001), unfavorable baseline hearing capability (p <0.0001), green-eyed patients (p <0.0001), and older age (p <0.0001). Using multivariate analysis, a prediction of individual hearing capabiltity after treatment was made. CONCLUSION: RT-induced hearing loss in the mean population is modest. However, clinically significant hearing loss was observed in older patients with green eyes and unfavorable pretreatment hearing. In these patients, the intended radiation dose may be adjusted according to the proposed predictive model, aiming to decrease the risk of ototoxicity. PMID- 19135317 TI - Three-staged stereotactic radiotherapy without whole brain irradiation for large metastatic brain tumors. AB - PURPOSE: To evaluate the efficacy and toxicity of staged stereotactic radiotherapy with a 2-week interfraction interval for unresectable brain metastases more than 10 cm(3) in volume. PATIENTS AND METHODS: Subjects included 43 patients (24 men and 19 women), ranging in age from 41 to 84 years, who had large brain metastases (> 10 cc in volume). Primary tumors were in the colon in 14 patients, lung in 12, breast in 11, and other in 6. The peripheral dose was 10 Gy in three fractions. The interval between fractions was 2 weeks. The mean tumor volume before treatment was 17.6 +/- 6.3 cm(3) (mean +/- SD). Mean follow-up interval was 7.8 months. The local tumor control rate, as well as overall, neurological, and qualitative survivals, were calculated using the Kaplan-Meier method. RESULTS: At the time of the second and third fractions, mean tumor volumes were 14.3 +/- 6.5 (18.8% reduction) and 10.6 +/- 6.1 cm(3) (39.8% reduction), respectively, showing significant reductions. The median overall survival period was 8.8 months. Neurological and qualitative survivals at 12 months were 81.8% and 76.2%, respectively. Local tumor control rates were 89.8% and 75.9% at 6 and 12 months, respectively. Tumor recurrence-free and symptomatic edema-free rates at 12 months were 80.7% and 84.4%, respectively. CONCLUSIONS: The 2-week interval allowed significant reduction of the treatment volume. Our results suggest staged stereotactic radiotherapy using our protocol to be a possible alternative for treating large brain metastases. PMID- 19135316 TI - Comparing digital tomosynthesis to cone-beam CT for position verification in patients undergoing partial breast irradiation. AB - PURPOSE: To evaluate digital tomosynthesis (DTS) technology for daily positioning of patients receiving accelerated partial breast irradiation (APBI) and to compare the positioning accuracy of DTS to three-dimensional cone-beam computed tomography (CBCT). METHODS AND MATERIALS: Ten patients who underwent APBI were scanned daily with on-board CBCT. A subset of the CBCT projections was used to reconstruct a stack of DTS image slices. To optimize soft-tissue visibility, the DTS images were reconstructed in oblique directions so that the tumor bed, breast tissue, ribs, and lungs were well separated. Coronal and sagittal DTS images were also reconstructed. Translational shifts of DTS images were obtained on different days from the same patients and were compared with the translational shifts of corresponding CBCT images. Seventy-seven CBCT scans and 291 DTS scans were obtained from nine evaluable patients. RESULTS: Tumor beds were best visible in the oblique DTS scans. One-dimensional positioning differences between DTS and CBCT images were 0.8-1.7 mm for the six patients with clips present and 1.2-2.0 mm for the three patients without clips. Because of the limited DTS scan angle, the DTS registration accuracy along the off-plane direction is lower than the accuracy along the in-plane directions. CONCLUSIONS: For patients receiving APBI, DTS localization offers comparable accuracy to CBCT localization for daily patient positioning while reducing mechanical constraints and imaging dose. PMID- 19135318 TI - [Uterine carcinosarcomas associated with tamoxifen therapy. Report of two cases and review of the literature]. AB - Carcinosarcomas are rare uterine cancers and carry poor prognosis. Although these tumours usually arise de novo, some cases developed under tamoxifen therapy have been reported. We report two more cases of uterine carcinosarcoma occurring in two postmenopausal patients benefiting from tamoxifen therapy as adjuvant treatment of breast cancer. A review of the literature is undertaken. PMID- 19135320 TI - Dogs cloned from fetal fibroblasts by nuclear transfer. AB - Fetal fibroblasts have been considered as the prime candidate donor cells for the canine reproductive cloning by somatic cell nuclear transfer (SCNT) in regard to the future production of transgenic dogs, mainly due to their higher developmental competence and handling advantage in gene targeting. In this study, the cloning efficiency with canine fetal fibroblasts as donor cells was determined. A total of 50 presumptive cloned embryos were reconstructed, activated and transferred into the oviducts of naturally synchronous recipient bitches. While the fusion rate (76.9%) was similar to those of our earlier studies with adult fibroblasts as donor cells (73.9-77.1%), a high cloning efficiency (4.0%; 2 births/50 embryos transferred) was found compared to the previous success rate with adult fibroblasts (0.2-1.8%). The cloned beagles were healthy and genotypically identical to the donor fibroblast cells. This study shows that a fetal fibroblast cell would be an excellent donor for future production of transgenic dogs via gene targeting in this cell followed cloning using SCNT technology. PMID- 19135319 TI - Sex heterogeneity in pharmacogenetic smoking cessation clinical trials. AB - Approximately one-quarter of smokers who use treatments for nicotine dependence are able to achieve cessation. However, there is evidence that women do not respond as well to nicotine replacement therapy (NRT) and, perhaps, to bupropion, compared to men. In this contribution to the Special Issue of Drug and Alcohol Dependence concerning Women and Smoking, we begin with a brief overview of data supporting the role of sex in influencing response to NRT and bupropion. Next, we summarize the results of pharmacogenetic smoking cessation clinical trials which assessed sex as a moderator as well. A relatively small number of pharmacogenetic studies of nicotine dependence treatments have been conducted and five studies reported sex effects in these trials. Of these trials, sex moderated the association of genetic variation in drug pharmacokinetics or pharmacodynamics and treatment response. We conclude this paper with a summary and a brief discussion of the major caveats of this literature and priorities for future research. PMID- 19135321 TI - Effects of cholesterol and cAMP on progesterone production in cultured luteal cells isolated from pseudopregnant cat ovaries. AB - The present study was designed to incubate luteal cells isolated from pseudopregnant cats and to investigate the effects of cholesterol and cAMP on luteal progesterone production. Corpora lutea were collected from the cats on days 10 and 15 of pseudopregnancy. Luteal cells were isolated from the ovaries by collagenase digestion. Steroidogenic luteal cells were stained for 3beta hydroxysteroid dehydrogenase (3beta-HSD) activity. Cells (2 x 10(4)) staining positive for 3beta-HSD were cultured for up to 7 days. The cells were treated with 22(R)-hydroxycholesterol (22R-HC) and dibutyryl cyclic AMP (dbcAMP) on days 1, 3 and 7. Treatment of cells with 22R-HC resulted in a dose-dependent increase (p<0.001) in progesterone production. When 22R-HC was used at a concentration of 10 microg/ml, it resulted in 2.7- and 5.1-fold increases in progesterone production on days 3 and 5, respectively. When the dose was doubled (20 microg/ml), treated cells produced four times more progesterone on days 3 and 7, and three times more on day 5. By day 7, progesterone production increased up to 9.1 times more than the control. Incubation of cells with both concentrations of dbcAMP (0.1 mM and 1 mM) resulted in significant stimulations of progesterone on days 5 and 7 (p<0.001). However, on day 3, only higher doses of dbcAMP (1 mM) resulted in significant stimulation (p<0.05). Progesterone production was increased up to 2- and 2.9-fold of the control when cells were treated with lower concentration of dbcAMP (0.1 mM) on days 5 and 7, respectively. Incubation of cells with 1 mM concentrations of dbcAMP induced a 3.2-fold increase on day 5 and a 5-fold increase on day 7. In conclusion, a successful incubation was performed for long-life culturing of luteal cells collected from pseudopregnant cats. The method works well and allows for optimal growth and development of cells in the culture. The present study also demonstrated that incubating cat luteal cells with 22R-HC and dbcAMP induces a significant increase in luteal progesterone synthesis. PMID- 19135322 TI - Effect of exogenous administration of buffalo follicular fluid on follicular development, estrus response and luteal function in anoestrous goats (Capra hircus). AB - The effect of buffalo follicular fluid (buFF) on follicular development, estrus response and luteal function was investigated in anoestrous does. Treatment with buFF (18 ml/doe) had no significant effect on the number of antral follicles of all class categories during the period of administration. However, after cessation of buFF treatment, the number of total antral follicles increased significantly with time (P<0.003) as well as due to the treatment x time interaction (P<0.02), without any influence on follicle size. Injection of buFF also caused a marked increase (P<0.049) with time in the number of medium-sized follicles at cessation. Approximately 60 and 20% of buFF-treated anoestrous does showed behavioural and silent estrus, respectively, compared to none in the control. The mean interval between cessation of buFF treatment to onset of oestrus and oestrus duration was 67.0+/-18.5 and 17.0+/-3.6h, respectively. Corpus lutea size varied between 4.6 and 5.8 mm with an average diameter of 5.2+/ 0.3mm. Only 33.3% of does showed serum progesterone levels above 1 ng/ml, while the remainder (66.7%) had below 0.5 ng/ml. Our results indicate that exogenous administration of buFF causes enhanced follicular activity following cessation of treatment, which results in behavioural oestrus and corpus luteum (CL) development in anoestrous does. CL development and its function is, however, inadequate in buFF-treated anoestrous does. PMID- 19135323 TI - Guidelines for the diagnosis and management of osteoporosis in postmenopausal women and men from the age of 50 years in the UK. AB - In 1999 and 2000 the Royal College of Physicians published guidelines for the prevention and treatment of osteoporosis [Royal College of Physicians. Osteoporosis: clinical guidelines for the prevention and treatment. London: Royal College of Physicians; 1999; Royal College of Physicians and Bone and Tooth Society of Great Britain. Update on pharmacological interventions and an algorithm for management. London, UK: Royal College of Physicians; 2000.; Royal College of Physicians. Glucocorticoid-induced osteoporosis. Guidelines on prevention and treatment; Bone and Tooth Society of Great Britain, National Osteoporosis Society and Royal College of Physicians. London, UK: Royal College of Physicians; 2002]. Since then, there have been significant advances in the field of osteoporosis including the development of new techniques for measuring bone mineral density, improved methods of assessing fracture risk and new treatments that have been shown to significantly reduce the risk of fractures. Against this background, the National Osteoporosis Guideline Group (NOGG), in collaboration with many Societies in the UK, have updated the original guidelines [Royal College of Physicians, National Osteoporosis Guideline Group on behalf of the Bone Research Society, British Geriatrics Society, British Orthopaedic Association, British Society of Rheumatology, National Osteoporosis Society, Osteoporosis 2000, Osteoporosis Dorset, Primary Care Rheumatology Society, Society for Endocrinology. Osteoporosis. Clinical guideline for prevention and treatment, Executive Summary. University of Sheffield Press; 2008], a practical summary of which is detailed below. The management algorithms are underpinned by a health economic analysis applied to the epidemiology of fracture in the UK. PMID- 19135324 TI - Lymphoepithelial carcinoma of the maxillary sinus with orbital invasion. AB - Lymphoepithelial carcinoma (LEC) of the maxillary sinus is a very rare neoplasm that shares some characteristics with nasopharyngeal carcinoma. Epstein-Barr virus (EBV) has been reported to be associated with LEC located outside of the nasopharynx in Asian populations. A case report of a 64-year-old Asian female with right-sided cheek mass which was diagnosed preoperatively as a maxillary mass by paranasal sinus computed tomography scan is presented. Because maxillary sinus cancer with orbital invasion was considered, she underwent surgical removal of a mass. Final pathology revealed LEC, which is presented in our report. However, in situ hybridization assays returned negative results for Epstein-Barr virus. The patient underwent postoperative chemoradiotherapy, and has remained disease-free during 3-year follow-up. Although a few cases in the maxillary LEC have been reported, this is the first report describing the diagnosis and treatment of LEC occurred in the maxillary sinus. Surgical removal and adjuvant chemoradiotherapy may be useful in the treatment of more advanced maxillary LEC. PMID- 19135325 TI - Formation of biofilm by Haemophilus influenzae isolated from pediatric intractable otitis media. AB - OBJECTIVES: The aims of this study are to evaluate biofilm formation by nontypeable Haemophilus influenzae (NTHi) isolated from children with acute otitis media (AOM) and its relation with clinical outcome of the disease. METHODS: Biofilm formations by NTHi clinical isolates from pediatric AOM patients were evaluated by a crystal violet microtiter plate and a 98 well pin-replicator assay with a confocal laser scanning microscopy (CLSM). Optical density values of clinical isolates were compared with a positive control and the ratio of clinical isolates to a positive control was defined as biofilm formation index (BFI). RESULTS: 84.3% clinical isolates of NTHi were biofilm forming strains (BFI> or =0.4). The BFI represented the levels of biofilm formation and adherence on the surface. The identical strains isolated from both middle ear fluids (MEFs) and nasopharynx showed biofilm formation at the same level. The prevalence of biofilm forming isolates was significantly higher among the susceptible strains than resistant strains. The level of biofilm formation of NTHi isolated from AOM cases who was not improved by amoxicillin (AMPC) was significantly higher than that of NTHi isolated from AOM cases who was improved by AMPC. CONCLUSION: We clearly showed the biofilm formation of clinical NTHi isolates from AOM children. In addition, the biofilm formed by NTHi would play an important role in persistent or intractable clinical course of AOM as a result of lowered treatment efficacy of antibiotics. PMID- 19135326 TI - Management of psychiatric disorders and addictive behaviors in patients with viral hepatitis C in France. AB - INTRODUCTION: Guidelines concerning the management of psychiatric disorders and addictive behaviors in patients with chronic hepatitis C and the conditions of collaboration between hepatogastroenterologists, infectiologists, psychiatrists and psychologists have not been published. This has a negative influence on optimal therapeutic management of chronic hepatitis C virus (HCV) infection. The aim of this study was to describe the current clinical practices for ambulatory management of psychiatric disorders and addictions, and the influence of a possible psychiatric and/or psychological collaboration. PATIENTS AND METHODS: A retrospective survey was conducted among 101 clinicians treating patients with chronic hepatitis C. Data were collected from personal interviews with the managing clinicians and from the files of patients with chronic hepatitis C patients who presented psychiatric disorders. RESULTS: Analysis of the 101 interviews and 598 patient files showed that 19% of patients had not received an optimal treatment for their HCV infection because of a psychiatric problem, and that less than 50% of the managing clinicians were working in collaboration with a psychiatrist or a psychologist. In conclusion, lack of collaboration between hepatogastroenterologists and psychiatrists could be deleterious for the optimal treatment of HCV infected patients. Improvement is required. PMID- 19135327 TI - High correlation between quantitative ultrasound and DXA during 7 years of follow up. AB - Ultrasound is a quick, cheap and non-radiating device for assessing bone quality. We wanted to validate the method for clinical and epidemiological use. Eighty women, aged 53-73 years, with osteoporosis and/or fractures were followed repeatedly during 7 years. Quantitative ultrasound (QUS) measurements (LUNAR Achilles) were compared with bone mineral density (BMD) and bone mineral content (BMC) estimated by DXA (LUNAR) in regions of interest. Changes in the speed of sound, broadband ultrasound attenuation and stiffness were positively correlated with changes in BMD and BMC in all regions measured with DXA (r=0.20-0.53; p=0.09 to <0.0001). The QUS t-score at the left heel was positively correlated with the t-score at the right heel (r=0.90, p<0.0001). The DXA t-score of the left vs. the right femur was also positively correlated (r=0.72-0.86; p<0.0001). A t-score< 2.5 S.D. was found in 70% and 56% at baseline, and 74% and 65% at follow-up measured with QUS and DXA, respectively. The mean sensitivity of QUS vs. DXA was 79% and the mean specificity 45% over a 7-year period. A QUS t-score of <-3.65 S.D. was consistent with a DXA t-score of <-2.5 S.D. In conclusion, QUS was well correlated with DXA in all regions over the 7-year period. QUS can be used in settings without access to DXA and in epidemiological studies. The sensitivity was high but the specificity was low, implicating that DXA, if available, is recommended before treatment for osteoporosis. However, treatment can be started without DXA at a QUS t-score<-3.65 S.D., and especially in the presence of fractures. PMID- 19135328 TI - The development and validation of a sensitive, dual-flow cell, SPR-based biosensor immunoassay for the detection, semi-quantitation, and characterization of antibodies to darbepoetin alfa and epoetin alfa in human serum. AB - A surface plasmon resonance (SPR)-based biosensor immunoassay was developed and validated using the Biacore 3000 instrument to detect, semi-quantitate, and characterize serum antibodies against darbepoetin alfa (Aranesp) and epoetin alfa (EPOGEN). In this sensitive, dual-flow cell assay, epoetin alfa and darbepoetin alfa are covalently immobilized onto consecutive flow cells of a carboxymethyl dextran-coated sensor chip. Diluted human serum samples are injected sequentially over both surfaces. The binding of serum antibodies to the immobilized proteins are detected and recorded in real time based on the principles of SPR. Furthermore, antibody binding is confirmed with a secondary anti-human immunoglobulin antibody. Positive samples are further characterized to determine the relative concentration of the antibodies using an affinity-purified, rabbit anti-epoetin alfa antibody as a reference control. The assay can detect 80ng/ml and 100ng/ml of antibody to epoetin alfa and darbepoetin alfa, respectively. The dynamic range of the assay is from 0.078microg/ml to 10microg/ml using a rabbit antibody with demonstrated accuracy and intra- and inter-assay precision. Approximately 80 serum samples can be analyzed on each sensor chip while maintaining a stable baseline and consistent immunological reactivity. The analysis of serum samples from subjects administered with epoetin alfa or darbepoetin alfa provided evidence that the assay can detect varying concentrations of antibodies of different off rates, isotypes, and IgG subclasses. PMID- 19135329 TI - Development of a high performance liquid chromatography method for quantification of PAC-1 in rat plasma. AB - A sensitive and specific high performance liquid chromatography method with UV detection was developed and validated for the determination of PAC-1 in rat plasma. After extraction with ethyl acetate, the chromatographic separation was carried out on a Diamonsil C(18) column (150mmx4.6mm i.d., 5microm particle size, Zhonghuida) protected by a ODS guard column (10mmx4.6mm i.d., 5microm particle size), using acetonitrile-methanol-phosphate buffer (pH 3.0, 30mM) (31:3:66, v/v/v) as mobile phase at a flow rate of 1.0mL/min, and wavelength of the UV detector was set at 281nm. No interference from any endogenous substances was observed during the elution of PAC-1 and internal standard (IS, indapamide). The calibration curve was linear over the range of 0.05-20microg/mL (r>0.99). The lower limit of quantification was evaluated to be 50ng/mL. The method was successfully applied to the pharmacokinetic study of PAC-1 after intravenous and oral administration in rats, respectively. PMID- 19135331 TI - Microbiologic study of the pathogens isolated from wound culture among Wenchuan earthquake survivors. AB - On May 12, an unprecedented earthquake struck Wenchuan County, Sichuan, China, and 1823 victims were admitted to West China Hospital, Sichuan, China. A total of 464 nonduplicate clinical isolates from wounds of earthquake victims were analyzed. The results show that the most common pathogen was Staphylococcus aureus, but only 24.4% of the total isolates were Gram-positive bacteria, and 73.2% were Gram-negative bacteria. The isolates were significantly different from isolation pattern of 2004 tsunami in Thailand. The isolation rates of extended spectrum beta-lactamase producer and pandrug-resistant Acinetobacter baumannii in this study might increase the risk of nosocomial infection. In this situation, clinical microbiologists, infection control staff, and administration decision makers should pay high attention to prevent disaster-associated nosocomial infections. PMID- 19135330 TI - Impact of AmpC overexpression on outcomes of patients with Pseudomonas aeruginosa bacteremia. AB - AmpC overexpression (AmpC++) is a significant mechanism of beta-lactam resistance in Pseudomonas aeruginosa, but its impact on clinical outcomes is not well established. To examine the influence of AmpC++ on clinical outcomes of patients with P. aeruginosa bacteremia, we screened all bloodstream P. aeruginosa isolates obtained from 2003 to 2006 for AmpC++. Demographics and outcomes were retrospectively compared between patients with P. aeruginosa bacteremia caused by AmpC++ and pan-susceptible strains (wild-type controls). Of the 263 isolates screened, 63 (24.0%) were nonsusceptible to ceftazidime. Clinical data of 42 AmpC++ isolates from 21 patients were compared with 33 control patients. The 2 groups were similar in sex and race. Patients in the AmpC++ group was more likely to receive inappropriate empiric antibiotics (odds ratio [OR] = 67.5; 95% confidence interval [CI], 6.3-720.0) and experience microbiologic persistence (OR = 12.2; 95% CI, 1.7-87.7). In institutions with a high prevalence of AmpC++, empiric therapy with agents with activity against AmpC++ strains may be warranted. PMID- 19135332 TI - [Pubalgia in athlete treated by atorvastatin]. PMID- 19135333 TI - [Management of coronary risk patients before anesthesia]. AB - Noninvasive stress tests such as myocardial perfusion scintigraphy or dobutamine stress echography have to be performed, only if they modify the perioperative strategy. Prophylactic pre-operative coronary revascularization can be helpful only in selected cases, independently from the operative context. Coronary stenting in patients undergoing noncardiac surgery (bare metal stent or drug eluting stents) adds an additional risk related to antiplatelet therapy management. Preoperative medical therapy optimisation is based on betablockers, and probably statins. Perioperative strategy in risk patients undergoing surgery is multimodal, it includes an adequate preoperative assessment, an optimisation of preoperative medical treatment, an optimisation of the per and post operative myocardial oxygen supply/requirement equilibrium. In case of low risk surgery, or in case of a patient with a good functional capacity, without symptoms, the operative assessment has to be minimal. For patients under antiplatelet therapy, a multidisciplinary discussion of optimal patient management strategy has to be notified in the patient file, this has to include the haemorrhagic and thrombosis discussion. PMID- 19135334 TI - [Clinical aspects and management of adult patients with acute gastroenteritis in primary care]. AB - AIM: In patients with acute gastroenteritis, general behaviour of patients as well as medical care remain poorly documented. The aim of this work was to determine clinical and demographic characteristics, reason for visiting a doctor, and medical care in adult patients with acute gastroenteritis. METHODS: During the winter epidemic peak of acute diarrhea, 2,217 general practitioners filled in standardized clinical observations of 17,273 patients (mean age 40 yrs) with acute gastroenteritis. RESULTS: There were abdominal pain, nausea, and vomiting in respectively 72%, 53% and 43% of patients. Thirty one percent of patients had fever. Nausea and vomiting were more frequently observed in younger patients (15 25 yrs) than in older ones. More severe diarrhea more frequent nausea, vomiting and abdominal pain (p<0.0001). Time for consulting was less than 24 and 48h in respectively 42% and 87% of patients, and was different according to professional classes. Predominant reason for consulting was the physical consequences of the disease (78%). Self medication was reported in 42% of patients and was different according to age and professional classes. Prescription of antidiarrheal drugs was reported by 91% of patients, sometimes in association with antiemetic (43%) and/or antispasmodic agents (35%). Additional explorations (mainly biological work-up) were rare but they were more frequently considered in older and female patients as well as in patients with severe and prolonged diarrhea. Stoppage of work was different among professional classes. It was reported in one third of patients, and was more frequent in patients with severe diarrhea. CONCLUSIONS: This study describes the behaviour of patients and doctors facing acute gastroenteritis and illustrates important fluctuations according age and professional classes. It shows high discomfort and repercussions of acute gastroenteritis especially regarding stoppage of work. PMID- 19135335 TI - [Charcot-Marie-Tooth disease]. AB - Charcot-Marie-Tooth (CMT) disease, also known as peroneal muscular atrophy or hereditary motor and sensory neuropathy, is among the most frequent hereditary disorders of the nervous system. The relatively homogeneous clinical phenotype involves mainly progressive weakness and wasting of distal muscles; it starts and predominates in the peroneal muscles. Electrophysiological and pathology data distinguish two principal forms of CMT: demyelinating and axonal. More than 20 distinct genetic subtypes have been identified to date and other new loci and genes remain to be discovered, thus demonstrating wide genetic heterogeneity and a number of different pathophysiological mechanisms. The classification of these different forms is based on both the mode of inheritance--autosomal dominant, recessive or X-linked--and the neuropathy type--demyelinating or axonal or "intermediate". The principal dominant forms are CMT1A, due to a duplication or point mutation in the PMP22 gene, and CMTX, due to mutations in the connexin 32 gene. Autosomal recessive forms are more frequent in North Africa. The most common involve mutations of GDAP1 or lamin A/C and generally lead to more severe phenotypes than the dominant forms. The great genetic heterogeneity necessitates a strategy for genetic diagnosis. It is based in part on the classification of the different genetic forms and in part on the phenotypic particularities and the frequency of the responsible genes in the population under study. PMID- 19135336 TI - Adventitial stripping for carotid sinus syndrome. AB - Patients with a carotid sinus syndrome (CSS) suffer from spells of dizziness and loss of conscience due to an exaggerated carotid baroreflex response. Three types of the syndrome are described: cardioinhibitory, vasodepressor and a mixed form. The gold standard for treatment is insertion of a pacemaker, but this therapy may be effective only in a pure cardioinhibitory type. In contrast, surgically interrupting afferent nerves at the origin may offer relief of symptoms in all three types. The present review analyzes the results of a surgical technique termed "adventitial stripping" of the carotid sinus. Data from 130 procedures demonstrate that carotid denervation is effective in 85% of CSS cases with low complication rates. Postoperative monitoring is mandatory as transient alterations in blood pressure and heart rate may occur. PMID- 19135337 TI - A note on variational Bayesian factor analysis. AB - Existing works on variational bayesian (VB) treatment for factor analysis (FA) model such as [Ghahramani, Z., & Beal, M. (2000). Variational inference for Bayesian mixture of factor analysers. In Advances in neural information proceeding systems. Cambridge, MA: MIT Press; Nielsen, F. B. (2004). Variational approach to factor analysis and related models. Master's thesis, The Institute of Informatics and Mathematical Modelling, Technical University of Denmark.] are found theoretically and empirically to suffer two problems: (1) penalize the model more heavily than BIC and (2) perform unsatisfactorily in low noise cases as redundant factors can not be effectively suppressed. A novel VB treatment is proposed in this paper to resolve the two problems and a simulation study is conducted to testify its improved performance over existing treatments. PMID- 19135338 TI - Assessing metabolic syndrome: waist circumference versus BMI. PMID- 19135339 TI - Prenatal infection and cavum septum pellucidum in adult schizophrenia. AB - Increased length of the cavum septum pellucidum (CSP) and in utero infection are each associated with increased risk of schizophrenia. Hence, we examined whether prenatal infections are related to CSP length in schizophrenia patients. In a well-characterized birth cohort, in utero infection was assessed using serologic biomarkers or physician diagnoses. Magnetic resonance images were acquired, and CSP length was quantified by a standard protocol. In utero infection was associated with increased CSP length in exposed schizophrenia cases compared to unexposed cases, suggesting that prenatal infection plays a role in a neurodevelopmental morphologic anomaly that has been related previously to schizophrenia. PMID- 19135340 TI - Configuration of bovine serum albumin adsorbed on polymer particles with grafted dextran corona. AB - The configuration of BSA macromolecules adsorbed on the surfaces of poly(alkylcyanoacrylate) nanoparticles has been determined using small angle neutron scattering (SANS). The nanoparticles were made by anionic emulsion polymerization (AEP) and self-assembly of dextran-poly(isobutylcyanoacrylate) (PICBA) copolymers. They have a hydrophobic PICBA core and a hydrophilic dextran corona. In vivo, they are recognized by the macrophages of the mononuclear phagocyte system. The amount of BSA bound to the particles, at adsorption equilibrium, has been determined through immunodiffusion, immunoelectrophoresis, and SANS. For particles with a radius of 25.3nm, the adsorption was found to saturate at 64 adsorbed BSA molecules per particle. The configuration of the adsorbed BSA molecules was determined from the SANS scattering curves, first at full contrast, and then at contrast match. Both experiments indicate that the BSA molecules are adsorbed on the PICBA core, in a flat configuration. This result may be important for understanding the in vivo opsonization mechanisms of nanoparticles and their resulting biodistribution. PMID- 19135341 TI - Alamethicin-lipid interaction studied by energy dispersive X-ray diffraction. AB - A detailed knowledge of the interaction between bacterial membranes and antibiotics provides important information to prevent high levels of antibiotic resistance exhibited by pathogenic strains. We investigated by energy dispersive X-ray diffraction (EDXD) the structure ordering of dioleoyl-phosphatidylcholine (DOPC) lipid interacting with antimicrobial peptide alamethicin, varying the lipid/peptide (L/P) molar ratio under two different hydration levels. In conditions of full hydration (100%) we found that the bilayer thickness is constant between L/P=20 and L/P=80 indicating that in this range, the system has reached the threshold value for the channel formation, while at the relative hydration of 45% a linear decrease of the bilayer thickness as function of L/P was revealed. The kinetic study of the complex alamethicin-DOPC at different L/P values, shows that the Bragg peak energy variation versus the hydration time has a biexponential behavior characterized by two different time constants. PMID- 19135342 TI - Methotrexate-incorporated polymeric nanoparticles of methoxy poly(ethylene glycol)-grafted chitosan. AB - We prepared methotrexate (MTX)-encapsulated polymeric nanoparticles using methoxy poly(ethylene glycol) (MPEG)-grafted chitosan (ChitoPEG) copolymer. MTX encapsulated polymeric nanoparticles of ChitoPEG copolymer has around 50-300nm in particle size and showed spherical shape when observed by transmission electron microscope (TEM). In (1)H nuclear magnetic resonance (NMR) study, the specific peaks of MTX and chitosan as a drug carrying inner-core disappeared at D(2)O and only the specific peak of MPEG was observed, while specific peaks of MPEG, MTX, and chitosan appeared in DCl/D(2)O mixtures. These results indicated that MTX was complexed with chitosan and then core-shell type nanoparticles had formed in aqueous environment, i.e., MTX/chitosan complexes composed of inner-core and MPEG composed of outer-shell of the nanoparticles. Loading efficiency of MTX in the polymeric nanoparticles was 94% (w/w) of ChitoPEG-1, 91.1% (w/w) of ChitoPEG-2, 90.1% (w/w) of ChitoPEG-3 and 65.2% (w/w) of ChitoPEG-4, expectively. The higher the drug feeding ratio, the higher the drug content and the lower the loading efficiency. The higher the MPEG graft ratio in the copolymer, the lower the drug content and loading efficiency. Drug contents evaluated by (1)H NMR were the same as found by UV spectrophotometer. PMID- 19135343 TI - Model of experts for decision support in the diagnosis of leukemia patients. AB - OBJECTIVE: Recent advances in the field of biomedicine, specifically in the field of genomics, have led to an increase in the information available for conducting expression analysis. Expression analysis is a technique used in transcriptomics, a branch of genomics that deals with the study of messenger ribonucleic acid (mRNA) and the extraction of information contained in the genes. This increase in information is reflected in the exon arrays, which require the use of new techniques in order to extract the information. The purpose of this study is to provide a tool based on a mixture of experts model that allows the analysis of the information contained in the exon arrays, from which automatic classifications for decision support in diagnoses of leukemia patients can be made. The proposed model integrates several cooperative algorithms characterized for their efficiency for data processing, filtering, classification and knowledge extraction. The Cancer Institute of the University of Salamanca is making an effort to develop tools to automate the evaluation of data and to facilitate de analysis of information. This proposal is a step forward in this direction and the first step toward the development of a mixture of experts tool that integrates different cognitive and statistical approaches to deal with the analysis of exon arrays. The mixture of experts model presented within this work provides great capacities for learning and adaptation to the characteristics of the problem in consideration, using novel algorithms in each of the stages of the analysis process that can be easily configured and combined, and provides results that notably improve those provided by the existing methods for exon arrays analysis. MATERIAL AND METHODS: The material used consists of data from exon arrays provided by the Cancer Institute that contain samples from leukemia patients. The methodology used consists of a system based on a mixture of experts. Each one of the experts incorporates novel artificial intelligence techniques that improve the process of carrying out various tasks such as pre processing, filtering, classification and extraction of knowledge. This article will detail the manner in which individual experts are combined so that together they generate a system capable of extracting knowledge, thus permitting patients to be classified in an automatic and efficient manner that is also comprehensible for medical personnel. RESULTS AND CONCLUSION: The system has been tested in a real setting and has been used for classifying patients who suffer from different forms of leukemia at various stages. Personnel from the Cancer Institute supervised and participated throughout the testing period. Preliminary results are promising, notably improving the results obtained with previously used tools. The medical staff from the Cancer Institute considers the tools that have been developed to be positive and very useful in a supporting capacity for carrying out their daily tasks. Additionally the mixture of experts supplies a tool for the extraction of necessary information in order to explain the associations that have been made in simple terms. That is, it permits the extraction of knowledge for each classification made and generalized in order to be used in subsequent classifications. This allows for a large amount of learning and adaptation within the proposed system. PMID- 19135344 TI - A review of recent methods for efficiently quantifying immunogold and other nanoparticles using TEM sections through cells, tissues and organs. AB - Detecting, localising and counting ultrasmall particles and nanoparticles in sub- and supra-cellular compartments are of considerable current interest in basic and applied research in biomedicine, bioscience and environmental science. For particles with sufficient contrast (e.g. colloidal gold, ferritin, heavy metal based nanoparticles), visualization requires the high resolutions achievable by transmission electron microscopy (TEM). Moreover, if particles can be counted, their spatial distributions can be subjected to statistical evaluation. Whatever the level of structural organisation, particle distributions can be compared between different compartments within a given structure (cell, tissue and organ) or between different sets of structures (in, say, control and experimental groups). Here, a portfolio of stereology-based methods for drawing such comparisons is presented. We recognise two main scenarios: (1) section surface localisation, in which particles, exemplified by antibody-conjugated colloidal gold particles or quantum dots, are distributed at the section surface during post-embedding immunolabelling, and (2) section volume localisation (or full section penetration), in which particles are contained within the cell or tissue prior to TEM fixation and embedding procedures. Whatever the study aim or hypothesis, the methods for quantifying particles rely on the same basic principles: (i) unbiased selection of specimens by multistage random sampling, (ii) unbiased estimation of particle number and compartment size using stereological test probes (points, lines, areas and volumes), and (iii) statistical testing of an appropriate null hypothesis. To compare different groups of cells or organs, a simple and efficient approach is to compare the observed distributions of raw particle counts by a combined contingency table and chi-squared analysis. Compartmental chi-squared values making substantial contributions to total chi-squared values help identify where the main differences between distributions reside. Distributions between compartments in, say, a given cell type, can be compared using a relative labelling index (RLI) or relative deposition index (RDI) combined with a chi-squared analysis to test whether or not particles preferentially locate in certain compartments. This approach is ideally suited to analysing particles located in volume-occupying compartments (organelles or tissue spaces) or surface-occupying compartments (membranes) and expected distributions can be generated by the stereological devices of point, intersection and particle counting. Labelling efficiencies (number of gold particles per antigen molecule) in immunocytochemical studies can be determined if suitable calibration methods (e.g. biochemical assays of golds per membrane surface or per cell) are available. In addition to relative quantification for between-group and between-compartment comparisons, stereological methods also permit absolute quantification, e.g. total volumes, surfaces and numbers of structures per cell. Here, the utility, limitations and recent applications of these methods are reviewed. PMID- 19135345 TI - Effect of parenteral or oral vinpocetine on the hemorheological parameters of patients with chronic cerebrovascular diseases. AB - INTRODUCTION: Hemorheological factors play an important role in the pathomechanism of ischemic cerebrovascular disorders. Abnormal rheological conditions in patients with chronic cerebrovascular disease predispose for recurrent strokes. Vinpocetine (VP), a synthetic ethyl esther of apovincamine, has successfully been used in the treatment of cerebrovascular diseases, in part because of its favourable rheological effects. PATIENTS AND METHODS: The study investigates the hemorheological changes in 40 patients in the chronic stage of ischemic cardiovascular disease after administration of vinpocetine. All patients received a high dose of intravenous VP in doses gradually increased to l mg/kg/day. In addition, 20 patients (mean age: 61+/-8 years) received 30 mg VP orally for 3 months. The other 20 patients (mean age: 59+/-6 years), who received placebo tablets, served as controls. Hemorheological parameters (hematocrit, plasma fibrinogen, whole blood viscosity, red blood cell aggregation and deformability) were evaluated at 1 and 3 months. RESULTS: The high-dose parenteral VP significantly decreased red blood cell aggregation, plasma and whole blood viscosity (p < 0.05) compared to the initial values. In patients with additional oral treatment, plasma and whole blood viscosities were significantly lower compared to the placebo patients at 3 months (p < 0.05). CONCLUSION: Our results confirmed the beneficial rheological effects of high-dose parenteral VP (partially caused by hemodilution) observed previously, and also warrant its long term oral admission to maintain the beneficial rheological changes. PMID- 19135346 TI - Madecassoside attenuates inflammatory response on collagen-induced arthritis in DBA/1 mice. AB - Madecassoside (MA), a triterpenoid product isolated from Centella asiatica, has been described to exhibit antioxidant and anti-inflammatory activities. The present study was undertaken to determine whether madecassoside (MA) is efficacious against collagen-induced arthritis (CIA) in mice and its possible mechanisms. DBA/1J mice were immunized with bovine type II collagen and treated with MA (3, 10 and 30 mg/kg d, i.g.) from days 21 to 42 after immunization. Arthritis was evaluated by hind paw swelling, polyarthritis index, and histological examination. In vitro proliferation of spleen cells was examined using 3-[4,5-dimethylthylthiazol-2-yl]-2, 5-diphenyltetrazoliumbromide (MTT) assay. Plasma levels of cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-10 (IL-10) and the expression of prostaglandin E(2) (PGE(2)), cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in synovial tissues were also determined. The results showed that comparing with untreated CIA mice, treated with MA dose-dependently suppressed the clinical arthritis score and joints tissues pathological damage, reduced the proliferation of spleen cells, plasma levels of TNF-alpha and IL-6, synovial tissues PGE(2) production and COX-2 protein expression, however, the expression of COX-1 in synovial tissues did not change and the plasma levels of IL-10 were increased. These results suggest that MA can effectively alleviate inflammatory response on CIA, and anti-inflammatory effects of MA can be attributed, at least partially, to the inhibition of pro-inflammatory mediators, including COX-2 expression, PGE(2) production, TNF-alpha and IL-6 levels and the up-regulation anti-inflammatory molecule IL-10. PMID- 19135348 TI - Randomized, placebo-controlled study of a selective PDE4 inhibitor in the treatment of asthma. AB - BACKGROUND: Phosphodiesterase-4 (PDE4) inhibitors have potential utility as a new therapeutic approach to improving symptoms and pulmonary function in asthma and COPD. This study evaluated the efficacy and safety of MK-0359, a selective and potent oral PDE4 inhibitor, in chronic asthma. METHODS: Adults (N=88) with > or =1 year asthma history and an FEV(1) 50-80% predicted were randomized to double blind treatment with MK-0359 (15mg/day) or placebo for 14 days, then crossed-over to the other treatment for 14 days. The primary endpoint was the change from baseline in FEV(1) at the end of each 2-week treatment period. Secondary and other endpoints included the changes from baseline in Daytime asthma symptom score, Nighttime asthma symptom score, Total daily beta-agonist use (puffs/day), AM and PM peak expiratory flow (PEF) and overall asthma-specific quality-of-life. Safety and tolerability were assessed by clinical adverse experiences. RESULTS: MK-0359 significantly improved the primary endpoint (versus placebo): the least squares mean difference in change from baseline in FEV(1) (L) was 0.09L (95% CI 0.01, 0.18). Endpoints of Daytime asthma symptom score, Nighttime asthma symptom score, Total daily beta-agonist use, AM PEF, PM PEF, and quality-of-life were also significantly improved. Nineteen patients (24.1%) on MK-0359 and 8 patients (10.4%) on placebo reported gastrointestinal clinical adverse experiences. Serious gastrointestinal clinical adverse experiences were reported in 3 patients while receiving MK-0359. CONCLUSION: Over a 14-day treatment period, the oral PDE4 inhibitor MK-0359 improved lower airway function, symptoms and rescue medication use in chronic asthma, although at the expense of gastrointestinal adverse experiences. (Clinical trial registry number: NCT00482898.). PMID- 19135349 TI - Limitation of the concentration of organic pollutants in sewage sludge for agricultural purposes: A case study in South Spain. AB - In 2000, the EU published the third draft of a future sludge directive entitled "Working document on sludge" where limit values for some organic compounds, including di-(2-ethyhexyl)phthalate (DEHP), sum of nonylphenol (NP), nonyphenol mono-(NP1EO) and diethoxylates (NP2EO), seven polychlorinated biphenyl congeners (PCB), polycyclic aromatic hydrocarbons (PAH) and linear alkylbenzene sulphonates (LAS), are fixed. In the present work, the monitoring of these organic compounds in sludge samples from four wastewater treatment plants (WWTPs) is reported. All WWTPs use anaerobic biological stabilization of sludge. The highest concentration levels were found for LAS, NPE and DEHP, in this order, with, in general, anaerobically-digested dehydrated sludge and compost samples being the most contaminated samples. DEHP, NPE, LAS and PAH were found at concentration levels above the limit values fixed in the third draft of the future EU sludge directive in the 44%, 88%, 13% and 6% of the analyzed anaerobically-digested dehydrated sludge and compost samples. PMID- 19135347 TI - Inhibitions of vascular endothelial growth factor expression and foam cell formation by EGb 761, a special extract of Ginkgo biloba, in oxidatively modified low-density lipoprotein-induced human THP-1 monocytes cells. AB - It has been reported that oxidatively modified low-density lipoprotein (Ox-LDL) involvement with vascular endothelial growth factor (VEGF) and foam cell formation play an important role in atherosclerosis (AS). Protective effects of Ginkgo biloba extract (EGb 761) have been identified for some cardiovascular and neurological disorders. The aim of this study was to investigate whether Ox-LDL regulates VEGF expression in human THP-1 monocytes, as well as the effect of EGb 761 on VEGF expression and the formation of foam cells. After exposure to Ox-LDL alone or in combination with EGb 761 for up to 48h, cell viability was measured using the MTT assay. VEGF protein content in the supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). VEGF mRNA was determined by real-time PCR. To determine the effect of EGb 761 on foam cell formation, an Ox-LDL-induced foam cell model was used. Ox-LDL inhibited the growth of THP-1 cells and EGb 761 increased the cell survival rate. Ox-LDL markedly increased VEGF expression in THP-1 cells in a time- and concentration-dependent manner, which was significantly suppressed by EGb 761. EGb 761 also inhibited monocyte/macrophage derived foam cell formation. These results suggest that Ox-LDL is involved in the development of human AS through VEGF induction in monocytes, and that EGb 761 prevents in vitro atherogenesis, probably via downregulation of VEGF expression in monocytes and inhibition of monocyte/macrophage-derived foam cell formation. The findings suggest a mechanism for the in vivo anti-AS effect of EGb 761 and support its potential clinical use in AS. PMID- 19135350 TI - EQCM immunoassay for phosphorylated acetylcholinesterase as a biomarker for organophosphate exposures based on selective zirconia adsorption and enzyme catalytic precipitation. AB - A zirconia (ZrO(2)) adsorption-based immunoassay by electrochemical quartz crystal microbalance (EQCM) has been initially developed, aiming at the detection of phosphorylated acetylcholinesterase (Phospho-AChE) as a potential biomarker for bio-monitoring exposures to organophosphate (OP) pesticides and chemical warfare agents. Hydroxyl-derivatized monolayer was preferably chosen to modify the crystal serving as the template for directing the electro-deposition of ZrO(2) film with uniform nanostructures. The resulting ZrO(2) film was utilized to selectively capture Phospho-AChE from the sample media. Horseradish peroxidase (HRP)-labeled anti-AChE antibodies were further employed to recognize the captured phosphorylated proteins. Enzyme-catalytic oxidation of the benzidine substrate resulted in the accumulation of insoluble product on the functionalized crystal. Ultrasensitive EQCM quantification by mass-amplified frequency responses as well as rapid qualification by visual color changes of product could be thus, achieved. Moreover, 4-chloro-1-naphthol (CN) was studied as an ideal chromogenic substrate for the enzyme-catalytic precipitation. Experimental results show that the developed EQCM technique can allow for the detection of Phospho-AChE in human plasma with a detection limit of 0.020 nM. Such an EQCM immunosensing format opens a new door towards the development of simple, sensitive, and field applicable biosensor for biologically monitoring low-level OP exposures. PMID- 19135351 TI - A novel automated discontinuous venous blood monitoring system for ex vivo glucose determination in humans. AB - Intensive insulin therapy reduces mortality and morbidity in critically ill patients but imposes great demands on medical staff who must take frequent blood samples for the determination of glucose levels. A solution to this resourcing problem would be provided by an automated blood monitoring system. The aim of the present clinical study was to evaluate such a system comprising an automatic blood sampling unit linked to a glucose biosensor. Our approach was to determine the correlation and system error of the sampling unit alone and of the combined system with respect to reference levels over 12h in humans. Two venous cannulae were inserted to connect the automatic and reference systems to the subjects. Blood samples were taken at 15 and 30 min intervals. The median Pearson coefficient of correlation between manually and automatically withdrawn blood samples was 0.982 for the sampling unit alone and 0.950 for the complete system. The biosensor had a linear range up to 20 mmoll(-1) and a 95% response time of <2 min. Clark Error Grid analysis showed that 96.93% of the data (228 data pairs) was in zone A and 3.07% in zone B. Insulin Titration Error Grid analysis suggested an acceptable treatment in 99.56% of cases. Implementation of a "Keep Vein Open" saline infusion into the automated blood sampling system reduced blood withdrawal failures through occluded catheters fourfold. In summary, automated blood sampling from a peripheral vein coupled with automatic glucose determination is a promising alternative to frequent manual blood sampling. PMID- 19135352 TI - Voltammetric studies on the HIV-1 inhibitory drug Efavirenz: the interaction between dsDNA and drug using electrochemical DNA biosensor and adsorptive stripping voltammetric determination on disposable pencil graphite electrode. AB - The interaction of Efavirenz (EFV) with fish sperm dsDNA immobilized onto pencil graphite electrode (PGE) has been studied by using differential pulse voltammetric technique using an electrochemical DNA biosensor. The guanine signal was lower with (double stranded-DNA) dsDNA-treated PGE than the untreated electrode after the interaction with EFV occurred. The changes in the experimental parameters such as the accumulation time and the concentration of EFV were also studied. All necessary parameters such as sensitivity, selectivity, accuracy and precision were calculated. In addition, the detection and determination limits, reproducibility and applicability of the analysis to pharmaceutical dosage forms were also investigated. These results showed that this DNA biosensor could be used for the sensitive, rapid simple and cost effective detection and determination of EFV-dsDNA interaction. The linearity was between 2 and 24 ppm of EFV concentration on guanine signal decreasing curve. EFV showed an irreversible oxidation behavior at all investigated pH values. This oxidation step was adsorption controlled on PGE. Hence, differential pulse adsorptive stripping (AdsDPV) voltammetric method was developed for the determination of EFV. Accumulation time and potential were optimized. Under these conditions, the current showed a linear dependence with concentration in the range between 0.018 and 2.56 ppm. Both determination methods were fully validated and applied for the analysis of EFV pharmaceutical dosage form. PMID- 19135353 TI - A novel method for the detection of point mutation in DNA using single-base-coded CdS nanoprobes. AB - A novel method for DNA point mutation detection using single-base-coded CdS nanoparticle probes is proposed. Target DNA was immobilized on core/shell Fe(3)O(4)/Au magnetic nanoparticles. Single-base-coded CdS nanoparticles, such as guanosine coded CdS (G-CdS), cytidine coded CdS (C-CdS), thymidine coded CdS (T CdS) and adenosine coded CdS (A-CdS) were used as the probes to identify the mutation sites in DNA strand. The hybridization process of single-base-coded CdS nanoparticle probes with the mutation sites in DNA was monitored using piezoelectric quartz crystal microbalance (QCM). The hybridization of the mutation base in DNA with its complementary base-coded CdS nanoprobes specifically caused significant changes in the resonance frequency of the QCM. Thus the base types of the mutation sites in DNA strand could be identified. The results were further confirmed by fluorescence measurement of CdS. Owing to its operation convenience and cost-effective, this DNA point mutation detection method is expected to hold a great promise in the detection of DNA point mutation and genetic assays. PMID- 19135354 TI - Bioelectrochemistry of hemoglobin immobilized on a sodium alginate-multiwall carbon nanotubes composite film. AB - This study described the bioelectrochemistry of hemoglobin (Hb) at multiwall carbon nanotubes (MWCNTs) noncovalently functionalized with biopolymers of sodium alginate (SA). The characteristics of Hb/SA-MWCNTs composite film were studied by using FT-IR spectroscopy, UV-vis spectroscopy and electrochemical methods. Hb immobilized on SA-MWCNTs composite film retained its native secondary structure, achieved direct electron transfer with the apparent heterogeneous electron transfer rate constant (k(s)) of (9.54+/-0.883) s(-1) and showed excellent bioelectrocatalytic activity to the reduction of hydrogen peroxide. The amperometric response of the biosensor varied linearly with the H(2)O(2) concentration ranging from 40 to 200 microM, with a detection limit of 16.41x10( 6) M (S/N=3) and the good long-term stability. Finally, we applied this proposed method to investigate the concentration of hydrogen peroxide in real samples. PMID- 19135355 TI - Label-free detection of ATP release from living astrocytes with high temporal resolution using carbon nanotube network. AB - Owing to its unique combination of electrical, physiochemical, and one-dimension structural properties, single-walled carbon nanotube (SWNT) has recently emerged as a novel nanoelectronic biosensor for biomolecular detection with extraordinary sensitivity and simple detection scheme. All the realizations so far, however, are limited to static in vitro measurement. Dynamic detection of biomolecule release from living cells which may occur in millisecond timescale has yet to be demonstrated. In the present work, SWNT network was utilized to directly interface with living neuroglial astrocytes and label-freely detect the triggered release of adenosine triphosphate (ATP) from these cells with high temporal resolution. The secreted ATP molecules diffuse into the narrow interface gap between the SWNT-net and the astrocyte, and interact with the nanotubes. Highly charged ATP molecules electrostatically modulate the SWNT conductance leading to measurable current response. This technique provides a novel platform to study ATP release and signaling which play important roles in astrocyte-neuron crosstalk and other essential cellular functions. PMID- 19135356 TI - Establishment of a chimeric reporting system for the universal detection and high throughput screening of G protein-coupled receptors. AB - G proteins, further divided into four subfamilies (G(s), G(q), G(12) and G(i)) based on their Galpha subunits, are the primary components activated by G protein coupled receptors (GPCRs). Current GPCR assays are limited to the evaluation of selective Galpha signaling and do not allow comprehensive screening for orphan GPCRs without a known coupled Galpha. Therefore, our aim was to design a chimeric reporting system that covers responses from all Galpha subfamilies simultaneously. Because G(s) activates cAMP response element (CRE)-driven genes whereas G(q) and G(12) activate serum response element (SRE)-driven genes, we therefore incorporated 2x CRE and 5x SRE (2CRE5SRE) into a promoter for driving luciferase expression. To further report G(i) signals, a 2CRE5SRE-driven chimeric G(qi), in which the C-terminus of G(q) is replaced by that of G(i), was integrated to switch the responses of G(i)-coupled GPCRs to the G(q) signaling. The novel reporter system showed a strong signal amplification when activated by neuromedin U receptor 1 (mainly activates G(q)), neuromedin U receptor 2 (mainly activates G(i)) or luteinizing hormone receptor (mainly through the G(s) and G(q) pathways). In addition, 293T cells stably carrying our reporter construct showed a similar sensitivity to the radioactive cAMP assay when revealing the constitutive signal from gain-of-function mutants of luteinizing hormone receptor. To our knowledge, this is the first reporting system capable of covering the G(s), G(q), G(12) and G(i) signals and revealing the phenomena of constitutively active GPCRs. Such a universal platform will benefit future high throughput screening and drug designs for any GPCR. PMID- 19135357 TI - Genomic hotspots of adaptation in butterfly wing pattern evolution. AB - What is the genetic architecture of morphological evolution? Is there uniform potential for novelty across a genome or, on the contrary, can a small number of large-effect genes explain the phenotypic variation observed within and between species? Here we highlight recent work on butterfly wing pattern genetics showing that a small set of loci can be repeatedly involved in the evolution of complex traits. These loci behave as genomic hotspots for diversification because they underlie adaptive variation within and between species with both convergent and highly divergent wing patterns. These findings suggest that certain loci may be more likely than others to facilitate rapid evolutionary change. PMID- 19135359 TI - Safety and efficacy of a triple antiemetic combination with the NK-1 antagonist aprepitant in highly and moderately emetogenic multiple-day chemotherapy. AB - AIM OF THE STUDY: In multiple-day chemotherapy (MDC), the combination of a 5 HT(3)-antagonist plus dexamethasone is still a standard of care. The role of a NK 1-antagonist remains to be defined. PATIENTS AND METHODS: Seventy eight cancer patients undergoing multiple-day chemotherapy of high (HEC) or moderate (MEC) emetic risk received granisetron, dexamethasone plus aprepitant during chemotherapy. After the end of chemotherapy, aprepitant plus dexamethasone was given for another 2 days. Primary end-point was complete response (CR) in the overall phase (day 1 until 5 days after the end of chemotherapy). RESULTS: Thirty eight patients underwent HEC and 40 patients underwent MEC for a median of 3.5 days. CR was seen in 57.9% and 72.5% of patients receiving HEC and MEC, respectively. The tolerability of the aprepitant regimen over 5-7 days was comparable with a 3-day aprepitant regimen. CONCLUSIONS: This is the first report in MDC with a NK-1-antagonist containing antiemetic regimen showing a favourable safety profile with good antiemetic efficacy. PMID- 19135358 TI - Adolescents with cancer in Italy: entry into the national cooperative paediatric oncology group AIEOP trials. AB - PURPOSE: Survival of adolescents (15-19 years old) with cancer has shown less favourable improvement in comparison with survival rates for younger children and older adults. This might be partly explained by the relative lack of participation of adolescents in cooperative clinical protocols. METHODS: This analysis compares the number of 15- to 19-year-olds treated at the paediatric oncology centres affiliated to the AIEOP (and registered in the 'model 1.01') with the number of incident cases predicted in Italy based on incidence rates from the Italian network of cancer registries (AIRTum). RESULTS: By 2006, over 22,000 cases had been registered in the model 1.01, and 1743 of these were adolescents. The ratio of observed/expected (O/E) cases of 15- to 19-year-olds was 0.10 (as opposed to 0.77 for the 0-14-year-old children), and this ratio increased from 0.05 to 0.18 over three successive study periods (1989-1994, 1995 2000 and 2001-2006). Sarcomas were the neoplasms with the highest O/E ratios, with 0.28 and 0.43 for osteosarcoma and Ewing sarcoma and 0.33 and 0.39 for rhabdomyosarcoma and other soft-tissue sarcomas, respectively. In the period 1989 2006, 55% of the adolescents registered (versus 69% of the children) were enrolled in formal national trials. CONCLUSION: Our study confirms a lower referral of 15- to 19-year-old adolescents to paediatric oncology units and their under-representation in clinical trials, but we also observed a progressive improvement in this situation in recent years. PMID- 19135360 TI - Thermophilic treatment of bulk drug pharmaceutical industrial wastewaters by using hybrid up flow anaerobic sludge blanket reactor. AB - The hybrid up flow anaerobic sludge blanket reactor was evaluated for efficacy in reduction of chemical oxygen demand (COD) and biochemical oxygen demand (BOD) of bulk drug pharmaceutical wastewater under different operational conditions. The start-up of the reactor feed came entirely with glucose, applied at an organic loading rate (OLR) 1 kg COD/m(3) d. Then the reactor was studied at different OLRs ranging from 2 to 11 kg COD/m(3)d with pharmaceutical wastewater. The optimum OLR was found to be 9 kg COD/m(3)d, where we found 65-75% COD and 80-94% of BOD reduction with biogas production containing 60-70% of methane and specific methanogenic activity was 320 ml CH(4)/g-VSS d. By the characterization studies of effluent using GC-MS, the hazardous compounds like phenol, l,2-methoxy phenol, 2,4,6-trichloro phenol, dibutyl phthalate, 1-bromo naphthalene, carbamazepine and antipyrine were present. After the treatment, these compounds degraded almost completely except carbamazepine. Thermophilic methanothrix and methanosaetae like bacteria are present in the granular sludge. PMID- 19135361 TI - Bioethanol, biohydrogen and biogas production from wheat straw in a biorefinery concept. AB - The production of bioethanol, biohydrogen and biogas from wheat straw was investigated within a biorefinery framework. Initially, wheat straw was hydrothermally liberated to a cellulose rich fiber fraction and a hemicellulose rich liquid fraction (hydrolysate). Enzymatic hydrolysis and subsequent fermentation of cellulose yielded 0.41 g-ethanol/g-glucose, while dark fermentation of hydrolysate produced 178.0 ml-H(2)/g-sugars. The effluents from both bioethanol and biohydrogen processes were further used to produce methane with the yields of 0.324 and 0.381 m(3)/kg volatile solids (VS)(added), respectively. Additionally, evaluation of six different wheat straw-to-biofuel production scenaria showed that either use of wheat straw for biogas production or multi-fuel production were the energetically most efficient processes compared to production of mono-fuel such as bioethanol when fermenting C6 sugars alone. Thus, multiple biofuels production from wheat straw can increase the efficiency for material and energy and can presumably be more economical process for biomass utilization. PMID- 19135362 TI - Start-up of the Anammox process from the conventional activated sludge in a membrane bioreactor. AB - A lab-scale membrane bioreactor (MBR) was used to start-up the anaerobic ammonium oxidation (Anammox) process from the conventional activated sludge for 2 months. Results indicated the MBR could be a novel and suitable system for start-up of the Anammox process. The Anammox activity appeared after 16 days operation, and the average removal efficiencies of ammonia and nitrite were both over 90% in the end. A final specific Anammox activity of 0.35 g NH(4)(+)-N+NO(2)(-)-N (gVSS *d)( 1) was obtained. Fluorescence in situ hybridization (FISH) analysis confirmed the existence of Anammox bacteria and aerobic ammonia oxidizing bacteria. On the basis of results on MBR performance and FISH analysis, it was proposed that the start-up process was essentially a microbial community succession under man-made disturbance, and a climax community with Anammox bacteria as the dominant population was finally established. PMID- 19135363 TI - Integrated anaerobic/aerobic biological treatment for intensive swine production. AB - Manure processing could help farmers to effectively manage nitrogen (N) surplus load. Many pig farms have to treat wastewater. Piggery wastewater treatment is a complex challenge, due to the high COD and N concentrations and low C/N ratio. Anaerobic digestion (AD) could be a convenient pre-treatment, particularly from the energetic view point and farm income, but this causes further reduction of C/N ratio and makes denitrification difficult. N removal can only be obtained integrating anaerobic/aerobic treatment by taking into account the best use of electron donors. Experiences gained in Italy during development of integrated biological treatment approaches for swine manure, from bench to full scale, are reported in this paper. Solid/liquid separation as pre-treatment of raw manure is an efficient strategy to facilitate liquid fraction treatment without significantly lowering C/N ratio. In Italy, two full scale SBRs showed excellent efficiency and reliability. Current renewable energy policy and incentives makes economically attractive the application of AD to the separated solid fraction using high solid anaerobic digester (HSAD) technology. Economic evaluation showed that energy production can reduce costs up to 60%, making sustainable the overall treatment. PMID- 19135364 TI - 2-Aminoimidazoles inhibitors of TGF-beta receptor 1. AB - The 4-(5-fluoro-6-methyl-pyridin-2-yl)-5-quinoxalin-6-yl-1H-imidazol-2-ylamine 3 is a potent and selective inhibitor of TGF-betaR1. Substitution of the amino group of 3 typically led to a slight decrease in the affinity for the receptor and in TGF-beta-inducted PAI-luciferase reporter activity. However, 2 acetamidoimidazoles were identified as attractive candidates for further optimization as a result of their significant activity combined to their superior pharmacokinetic profile. PMID- 19135365 TI - Synthesis of Pro-Xylane: a new biologically active C-glycoside in aqueous media. AB - The scope and limitation of Lubineau's reaction were evaluated for the synthesis of C-glycosides (compounds 1-13). Further transformation of side chain carbonyl was also achieved (compounds 16-23). Optimization of these two steps was investigated in xylose case. Some of the compounds were shown to stimulate sulfated glycosaminoglycans (GAGs) synthesis. Compound 20 (called Pro-Xylane) was identified as the best activator of GAGs biosynthesis. Pro-Xylane was developed using environmentally friendly conditions relevant to 'Green-Chemistry' principles and launched on the market in September 2006. This compound is the first example of 'Green' chemical used in cosmetic. PMID- 19135366 TI - A motor component to the memories of habitual foraging routes in wood ants? AB - Ants are so low to the ground that slight undulations in the terrain over which they navigate will cause large and unpredictable changes to their view of the scene around them. We describe here evidence of a form of motor learning that helps ants follow their usual route when guiding landmarks vanish from sight. Wood ants were trained to approach a vertical bar presented at varying positions on a LCD screen. In different experiments, the bar was either stationary, moved smoothly, or jumped between two stationary positions. Ants trained in these three ways followed straight, curved, or two-leg routes, respectively. Once ants were accustomed to approaching the bar from different starting points, the bar was made to disappear during their approach. Ants often continued their straight or curved or two-leg paths, despite the missing landmark, showing that they can perform complex routes with no more than intermittent visual feedback. PMID- 19135367 TI - Reduction of the crowding effect in spatially adjacent but cortically remote visual stimuli. AB - When embedded in adjacent distractors, a target becomes more difficult to perceive. The neural mechanism for this ubiquitous visual crowding effect remains unresolved. Stimuli presented on opposite sides of the vertical meridian initially project to different hemispheres, whereas stimuli with the same spatial distance but presented to one side of the vertical meridian project to the same hemisphere. Dissociation between visual spatial distance and cortical distance can also be found in V2 and V3 (quadrant representations of the visual hemifield) along the horizontal meridian. In the current study, we observed a strong crowding effect from spatially adjacent distractors with either Gabor or letter targets presented near the vertical or horizontal meridian. Interestingly, for a target presented near the vertical meridian, a distractor from the same side of the meridian (cortically near) had a significantly stronger crowding effect compared with an equidistant distractor presented on the opposite side (cortically remote). No such meridian modulation was observed across the horizontal meridian. These results constrain the cortical locus of the crowding effect to a stage in which left and right visual spaces are represented discontinuously but the upper and lower visual fields are represented continuously, likely beyond the early retinotopic areas. PMID- 19135368 TI - Plant-specific protein MCD1 determines the site of chloroplast division in concert with bacteria-derived MinD. AB - Chloroplasts evolved from a cyanobacterial endosymbiont, and chloroplast division requires the formation of an FtsZ division ring, which is descended from the cytokinetic machinery of cyanobacteria. As in bacteria, the positioning of the chloroplast FtsZ ring is regulated by the proteins MinD and MinE. However, chloroplast division also involves mechanisms invented by the eukaryotic host cell. Here we show that a plant-specific protein MULTIPLE CHLOROPLAST DIVISION SITE 1 (MCD1) regulates FtsZ ring positioning in Arabidopsis thaliana chloroplasts. Our analyses show that both MCD1 and MinD are required for chloroplast division, localizing at the division sites and punctate structures dispersed on the inner envelope. MinD overexpression inhibited FtsZ ring formation whereas MCD1 overexpression did not. Localization studies suggest that MCD1 is required for MinD localization to regulate FtsZ ring formation. Furthermore, the interaction between MCD1 and MinD in yeast two-hybrid assays suggests that MCD1 recruits MinD by direct interaction. These results point out differences in the MinD localization mechanism between chloroplasts and bacterial model systems and suggest that the plant cell evolved a component to modulate the cyanobacteria-derived Min system so as to regulate chloroplast FtsZ ring positioning. PMID- 19135369 TI - Cuticular hydrocarbons reliably identify cheaters and allow enforcement of altruism in a social insect. AB - Cheaters are a threat to every society and therefore societies have established rules to punish these individuals in order to stabilize their social system. Recent models and observations suggest that enforcement of reproductive altruism (policing) in hymenopteran insect societies is a major force in maintaining high levels of cooperation. In order to be able to enforce altruism, reproductive cheaters need to be reliably identified. Strong correlational evidence indicates that cuticular hydrocarbons are the means of identifying cheaters, but direct proof is still missing. In the ant Aphaenogaster cockerelli, we mimicked reproductive cheaters by applying a synthetic compound typical of fertile individuals on nonreproductive workers. This treatment induced nestmate aggression in colonies where a queen was present. As expected, it failed to do so in colonies without a queen where workers had begun to reproduce. This provides the first direct evidence that cuticular hydrocarbons are the informational basis of policing behaviors, serving a major function in the regulation of reproduction in social insects. We suggest that even though cheaters would gain from suppressing these profiles, they are prevented from doing so through the mechanisms of hydrocarbon biosynthesis and its relation to reproductive physiology. Cheaters are identified through information that is inherently reliable. PMID- 19135370 TI - Distinctive Paleo-Indian migration routes from Beringia marked by two rare mtDNA haplogroups. AB - BACKGROUND: It is widely accepted that the ancestors of Native Americans arrived in the New World via Beringia approximately 10 to 30 thousand years ago (kya). However, the arrival time(s), number of expansion events, and migration routes into the Western Hemisphere remain controversial because linguistic, archaeological, and genetic evidence have not yet provided coherent answers. Notably, most of the genetic evidence has been acquired from the analysis of the common pan-American mitochondrial DNA (mtDNA) haplogroups. In this study, we have instead identified and analyzed mtDNAs belonging to two rare Native American haplogroups named D4h3 and X2a. RESULTS: Phylogeographic analyses at the highest level of molecular resolution (69 entire mitochondrial genomes) reveal that two almost concomitant paths of migration from Beringia led to the Paleo-Indian dispersal approximately 15-17 kya. Haplogroup D4h3 spread into the Americas along the Pacific coast, whereas X2a entered through the ice-free corridor between the Laurentide and Cordilleran ice sheets. The examination of an additional 276 entire mtDNA sequences provides similar entry times for all common Native American haplogroups, thus indicating at least a dual origin for Paleo- Indians. CONCLUSIONS: A dual origin for the first Americans is a striking novelty from the genetic point of view, and it makes plausible a scenario positing that within a rather short period of time, there may have been several entries into the Americas from a dynamically changing Beringian source. Moreover, this implies that most probably more than one language family was carried along with the Paleo Indians. PMID- 19135371 TI - Nursing involvement improves compliance with tight blood glucose control in the trauma ICU: a prospective observational study. AB - INTRODUCTION: The importance of tight glycaemic control has gained acceptance over the last 5 years as a critical component of routine intensive care unit (ICU) measures. In an environment already strained for resources and staffing, however, effective strategies providing for increased input and responsibility of bedside nursing personnel are paramount to successful implementation. HYPOTHESIS: Increasing input and responsibilities of ICU nursing staff in tight glycaemic control policies improves glucose control in the trauma ICU. METHODS: After Institutional Review Board approval, we conducted a prospective "before-after" trial examining the effect of nursing education and input on outcome of a tight (goal 80-120 mg/dL) glycaemic control protocol. After a three month assessment of compliance with a previously physician-developed protocol, an educational in service was conducted for all trauma ICU nursing staff. Nursing staff were then asked to provide input on the development of a new protocol using multiple-choice ballots to define 7 components of protocol criteria. Using nursing input, we developed and implemented a new glycaemic protocol that shifted much of the responsibility for initiation and subsequent adjustment of insulin infusion to the bedside nurse, allowing them to more liberally utilise their bedside clinical judgment and knowledge of the specific patient. RESULTS: Nursing input on seven factors of protocol criteria did not differ significantly from the previously existing protocol, except with reference to nursing desire for increased responsibility in the implementation and maintenance of tight glycaemic control. After three months implementation of a new protocol developed utilising nursing input, both mean blood glucose levels achieved (137.8 mg/dL vs. 128.2mg/dL, p=0.028) and time to first hourly blood glucose within goal range (<120 mg/dL) was improved (36 h vs. 9h). The number of hypoglycaemic (BS <60) episodes increased slightly after revision (1 event vs. 5 event), with no hypoglycaemic seizures or coma occurring during either period. CONCLUSION: Nursing input and increased responsibility improved the results of a tight glycaemic control in our trauma ICU. Increasing nursing input in the development and implementation of a tight glycaemic policies can result in safe and effective improved glucose control in the trauma ICU. PMID- 19135372 TI - Gait event detection using a multilayer neural network. AB - Manual detection of gait events via visual inspection of motion capture data is a laborious process. There are currently no robust techniques available to automate the process for pathologic gait. However, the detection of gait events is essentially a classification problem; an application for which artificial neural networks are well suited. In this paper, a multilayer artificial neural network is presented for the purpose of classifying foot-contact and foot-off events using the sagittal plane coordinates of heel and toe markers. The timing of events detected using this method was compared to the timing of events detected by measuring the ground reaction force using a force plate for a total of 40 pathologic subjects divided into two groups: barefoot and shod/braced. On average, the neural network detected foot-contact events 7.1 ms and 0.8 ms earlier than the force plate for the barefoot and shod/braced groups respectively. The average difference for foot-off events was 8.8 ms and 3.3 ms. Given that motion capture data were collected at 120 Hz, this implies that the force plate method and neural network method generally agreed within 1-2 frames of data. Consequently, the neural network was shown to be an accurate, autonomous method for detecting gait events in pathologic gait. PMID- 19135373 TI - Gait and clinical movement analysis research priorities: 2007 update from the research committee of the Gait and Clinical Movement Analysis Society. PMID- 19135374 TI - Protein S-glutathionylation: a regulatory device from bacteria to humans. AB - S-Glutathionylation is the specific post-translational modification of protein cysteine residues by the addition of the tripeptide glutathione, the most abundant and important low-molecular-mass thiol within most cell types. Protein S glutathionylation is promoted by oxidative or nitrosative stress but also occurs in unstressed cells. It can serve to regulate a variety of cellular processes by modulating protein function and to prevent irreversible oxidation of protein thiols. Recent findings support an essential role for S-glutathionylation in the control of cell-signalling pathways associated with viral infections and with tumour necrosis factor-(-induced apoptosis. Glyceraldehyde-3-phosphate dehydrogenase has recently been implicated in the regulation of endothelin-1 synthesis by a novel, S-glutathionylation-based mechanism involving messenger RNA stability. Moreover, recent studies have identified S-glutathionylation as a redox signalling mechanism in plants. PMID- 19135375 TI - Sub-vastus approach is more effective than a medial parapatellar approach in primary total knee arthroplasty: a randomized controlled trial. AB - In a prospective single-centre longitudinal randomized controlled trial 116 patients were allocated to the sub-vastus approach, and 115 to the medial parapatellar approach. At one week follow-up, compared to baseline, range of motion, Knee Society (KS) global, KS knee, and KS pain scores were significantly better in the sub-vastus group. At the one year follow-up, WOMAC global and pain scores, SF36 physical function and role-physical scores, and EuroQol utility and pain score were significantly better in the sub-vastus group. The ease of exposure in the sub-vastus approach was significantly worse. There was no significant difference in length of stay or analgesia intake. The sub-vastus approach to total knee arthroplasty was more effective than a medial parapatellar approach at both one week and one year post-operatively, but surgeons reported a less easy exposure in the sub-vastus group. PMID- 19135376 TI - Novel 6-methanesulfonamide-3,4-methylenedioxyphenyl-N-acylhydrazones: orally effective anti-inflammatory drug candidates. AB - We described herein the molecular design of novel in vivo anti-inflammatory 6 methanesulfonamide-3,4-methylenedioxyphenyl-N-acylhydrazone derivatives (1) planned by applying the molecular hybridization approach. This work also points out to the discovery of LASSBio-930 (1c) as a novel anti-inflammatory and anti hyperalgesic prototype, which was able to reduce carrageenan-induced rat paw edema with an ED(50) of 97.8 micromol/kg, acting mainly as a non-selective COX inhibitor. PMID- 19135377 TI - Non-vanillyl resiniferatoxin analogues as potent and metabolically stable transient receptor potential vanilloid 1 agonists. AB - A series of non-vanillyl resiniferatoxin analogues, having 4 methylsulfonylaminophenyl and fluorophenyl moieties as vanillyl surrogates, have been investigated as ligands for rat TRPV1 heterologously expressed in Chinese hamster ovary cells. Although lacking the metabolically problematic 4-hydroxy substituent on the A-region phenyl ring, the compounds retained substantial agonist potency. Indeed, the 3-methoxy-4-methylsulfonylaminophenyl analog (1) was modestly (2.5-fold) more potent than RTX, with an EC(50)=0.106 nM. Further, it resembled RTX in its kinetics and pattern of stimulation of the levels of intracellular calcium in individual cells, as revealed by imaging. Compound 1 displayed modestly enhanced in vitro stability in rat liver microsomes and in plasma, suggesting that it might be a pharmacokinetically more favorable surrogate of resiniferatoxin. Molecular modeling analyses with selected analogues provide evidence that the conformational differences could affect their binding affinities, especially for the ester versus amide at the B-region. PMID- 19135378 TI - Conformational analysis of trimeric maleimide substituted 1,5,9 triazacyclododecane HIV fusion scaffolds. AB - An analysis of the conformational preferences of three trimeric maleimide substituted 1,5,9-triazacyclododecane derivatives, proposed as cross linking reagents for HIV-1 fusion inhibitors, is presented. Exhaustive sampling was performed using the mixed Low Mode Monte Carlo conformational searching technique on the corresponding OPLS2005/GBSA(water) potential energy surface. Geometric structure, molecular length, and hydrogen bonding patterns of the compounds are analyzed. Global minimum energy structures were verified as minima using B3LYP/6 31G * geometry optimization. All structures adopt a crown-like 12-membered ring conformation; however, the system with the shortest maleimide arms (1a) can also adopt alternative ring orientations. Overall, derivatives with longer maleimide arms were more flexible and resulted in ensembles with a larger number of low energy structures. Comparison with biological inhibition data indicates that there is very little relationship between molecular size and the ability of the scaffold to orient CD4M9 miniproteins for optimal inhibition; however hydrophobicity may play a role. PMID- 19135379 TI - Synthesis of antisense oligonucleotides containing 2'-O-psoralenylmethoxyalkyl adenosine for photodynamic regulation of point mutations in RNA. AB - 2'-O-psoralen-conjugated antisense oligonucleotide was able to recognize a point mutation of mRNA. It had outstanding ability to photo-cross-link only to oligoribonucleotides (ORN) having a point mutation. This type of antisense molecule is the only one of its kind so far. To give high photo-cross-linking efficiency and sequence selectivity to antisense molecules, we synthesized novel photo-reactive oligonucleotides (2'-Ps-xom) containing psoralen at the 2'-O position adenosine via an ethoxymethylene (2'-Ps-eom), propoxymethylene (2'-Ps pom) and butoxymethylene (2'-Ps-bom) linker, respectively. We evaluated the photo cross-linking efficiency and sequence selectivity in photo-cross-linking of 2'-Ps xom to the complementary ORN and to an ORN having a mismatch base. Among them, 2' Ps-eom exhibited superior photo-cross-linking efficiency with high sequence selectivity. PMID- 19135380 TI - Solution- and solid-phase synthesis and anti-HIV activity of maslinic acid derivatives containing amino acids and peptides. AB - Maslinic acid (1) has been coupled at C-28 with several alpha- and omega-amino acids by using solution- and solid-phase synthetic procedures. Twelve derivatives (2-13) with a single amino acid residue were prepared in solution phase, whereas a dipeptide (14), a tripeptide (15), and a series of conjugate dipeptides (16-24) were synthesized in solid phase. The anti-HIV activity of these compounds was assessed on MT-2 cells infected with viral clones carrying the luciferase gene as a reporter. While in maslinic acid (1) were present both cytotoxic and antiviral activities, only the derivatives 13 and 24 showed anti-HIV-1 activity and therefore represent a novel class of anti-HIV-1 compounds. PMID- 19135381 TI - Cyclotron and linac production of Ac-225. AB - Radium needles that were once implanted into tumours as a cancer treatment are now obsolete and constitute a radioactive waste problem, as their half-life is 1600 years. The reduction of radium by photonuclear transmutation by bombarding Ra-226 with high-energy photons from a medical linear accelerator (linac) has been investigated. A linac dose of 2800 Gy produced about 2.4 MBq (64 microCi) of Ra-225, which decays to Ac-225 and can then be used for 'Targeted Alpha Therapy' (TAT) of cancer. This result, while consistent with theoretical calculations, is far too low to be of practical use unless much larger quantities of radium are irradiated. The increasing application of Ac-225 for cancer therapy indicates the potential need for its increased production and availability. This paper investigates the possibility of producing of Ac-225 in commercial quantities, which could potentially reduce obsolete radioactive material and displace the need for expensive importation of Ac-225 from the USA and Russia in the years ahead. Scaled up production of Ac-225 could theoretically be achieved by the use of a high current cyclotron or linac. Production specifications are determined for a linac in terms of current, pulse length and frequency, as well as an examination of other factors such as radiation issues and radionuclei separation. Yields are compared with those calculated for the Australian National Cyclotron in Sydney. PMID- 19135382 TI - Cavopulmonary anastomosis improves left ventricular assist device support in acute biventricular failure. AB - OBJECTIVE: Right ventricular failure during left ventricular assist device (LVAD) support can result in severe hemodynamic compromise with high mortality. This study investigated the acute effects of cavopulmonary anastomosis on right ventricular loading and LVAD performance in a model of severe biventricular failure. METHODS: LVAD support was performed by means of centrifugal pump implantation in 14 anesthetized dogs (20-30 kg) with severe biventricular failure obtained by ventricular fibrillation induction. Animals were randomized to be submitted to classical cavopulmonary anastomosis (Glenn shunt) or to control group and were maintained under LVAD support for 2h. Left and right atrial, right ventricular and systemic pressures were monitored, while total pulmonary flow was simultaneously recorded by transonic flowmeters located on the superior vena cava and pulmonary trunk. Blood gas and venous lactate determinations were also obtained. RESULTS: Ventricular fibrillation maintenance resulted in acute LVAD performance impairment after 90 min in the control group, while animals with Glenn circuit maintained normal LVAD pump flow (55+/-13 ml kg(-1)min(-1) vs 21+/ 4 ml kg(-1)min(-1), p<0.001) and better peripheral perfusion (blood lactate of 29+/-10 pg/ml vs 46+/-9 pg/ml, p<0.001). Left and right atrial pressures did not change significantly, while right ventricular pressure was lower in animals with Glenn circuit (13+/-3 mm Hg vs 22+/-8 mm Hg, p=0.005). Right ventricular unloading with Glenn shunt also resulted in superior total pulmonary flow (59+/ 13 ml kg(-1)min(-1) vs 17+/-3 ml kg(-1)min(-1), p<0.001). CONCLUSION: The concomitant use of cavopulmonary anastomosis during LVAD support in a model of severe biventricular failure limited right ventricular overloading and resulted in better hemodynamic performance. PMID- 19135383 TI - Effects of linear polarized infrared light irradiation on the transcriptional regulation of IL-8 expression in IL-1beta-stimulated human rheumatoid synoviocytes involves phosphorylation of the NF-kappaB RelA subunit. AB - Although recent clinical studies have shown that laser therapy acts as an anti inflammatory effector in the treatment of some diseases, little is known about the mechanism by which it acts in rheumatoid arthritis (RA) patients. The purpose of our work was to examine how irradiation with linear polarized infrared light (LPIL) suppresses inflammatory responses in the MH7A rheumatoid fibroblast-like synoviocyte cell line. We initially confirmed the effects of two disease modifying anti-rheumatic treatments, LPIL irradiation and dexamethasone (Dex) administration, under experimental inflammatory conditions using gene chip technology. We found that LPIL exerted a smaller effect on gene transcription than Dex; however, IL-1beta-inducible target genes such as the CXCL type chemokines IL-8, IL-1beta and IL-6 were all clearly suppressed by LPIL to the same degree as by Dex. We also found that IL-1beta-induced release of IL-8 from MH7A cells was completely blocked by pretreatment with the (IL-8) inhibitor Bay11 7085, indicating that activation of NF-kappaB signaling plays an important role in the secretion of IL-8. Although the levels of NFKB1 and RELA transcription were unaffected by IL-1beta stimulation, phosphorylation of RelA S276 was suppressed by both LPIL and Dex. Thus LPIL likely exerts its anti-inflammatory effects by inhibiting the release of the inflammatory chemokine IL-8. A fuller understanding of the anti-inflammatory mechanism of LPIL in rheumatoid synoviocytes could serve as the basis for improved treatment of RA patients in the future. PMID- 19135384 TI - Inflammation and vascular calcification in chronic kidney disease: the role of Fetuin-A. PMID- 19135385 TI - A novel approach to collision-induced dissociation (CID) for ion mobility-mass spectrometry experiments. AB - Collision induced dissociation (CID) combined with matrix assisted laser desorption ionization-ion mobility-mass spectrometry (MALDI-IM-MS) is described. In this approach, peptide ions are separated on the basis of mobility in a 15 cm drift cell. Following mobility separation, the ions exit the drift cell and enter a 5 cm vacuum interface with a high field region (up to 1000 V/cm) to undergo collisional activation. Ion transmission and ion kinetic energies in the interface are theoretically evaluated accounting for the pressure gradient, interface dimensions, and electric fields. Using this CID technique, we have successfully fragmented and sequenced a number of model peptide ions as well as peptide ions obtained by a tryptic digest. This instrument configuration allows for the simultaneous determination of peptide mass, peptide-ion sequence, and collision-cross section of MALDI-generated ions, providing information critical to the identification of unknown components in complex proteomic samples. PMID- 19135387 TI - Rantings of an associate editor: a plea to manuscript reviewers. PMID- 19135388 TI - Active pauses induce more variable electromyographic pattern of the trapezius muscle activity during computer work. AB - The aim of this laboratory study was to evaluate effects of active and passive pauses and investigate the distribution of the trapezius surface electromyographic (SEMG) activity during computer mouse work. Twelve healthy male subjects performed four sessions of computer work for 10 min in one day, with passive (relax) and active (30% maximum voluntary contraction of shoulder elevation) pauses given every 2 min at two different work paces (low/high). Bipolar SEMG from four parts of the trapezius muscle was recorded. The relative rest time was higher for the lower parts compared with the upper of the trapezius (p<0.01). The centroid of exposure variation analysis (EVA) along the time axis was lower during the computer work with active pause compared with passive one (p<0.05). The results of this study revealed (i) lower rest time for the upper parts of trapezius compared with the lower parts, in line with previous clinical findings, (ii) active pauses contributed to a more variable muscle activity pattern during computer work that might have functional implications with respect to work-related musculoskeletal disorders. PMID- 19135386 TI - HD desktop: an integrated platform for the analysis and visualization of H/D exchange data. AB - Here we describe an integrated software platform titled HD Desktop designed specifically to enhance the analysis of hydrogen/deuterium exchange (HDX) mass spectrometry data. HD Desktop integrates tools for data extraction with visualization components within a single web-based application. The interface design enables users to navigate from the peptide view to the sample and experiment levels, tracking all manipulations while updating the aggregate graphs in real time. HD Desktop is integrated with a relational database designed to provide performance enhancements, as well as a robust model for data storage and retrieval. Additional features of the software include retention time determination, which is achieved with the use of theoretical isotope fitting; here, we assume that the best theoretical fit will occur at the correct retention time for any given peptide. Peptide data consolidation for the rendering of data in 2D was realized by automating known and novel approaches. Designed to address broad needs of the HDX community, the platform presented here provides an efficient and manageable workflow for HDX data analysis and is freely available as a web tool at the project home page http://hdx.florida.scripps.edu. PMID- 19135389 TI - A Braunwald-Cutter valve: a mitral prosthesis at 33 years. AB - We present a case of a 76-year-old woman with a Braunwald-Cutter mitral caged ball valve prosthesis excised after 33 years post implantation due to a paravalvular leak. The valve itself was intact and fully functional. We believe the longevity of this valve was due to the decreased flow velocities and the lower pressure in the mitral valve position. PMID- 19135390 TI - Hemangioma of the right atrium: imaging and pathology. AB - BACKGROUND: Cardiac hemangiomas are benign neoplasms which have been reported to appear as well-circumscribed, homogenous, enhancing masses at imaging. METHODS AND RESULTS: We report a 49-year-old woman with a cardiac hemangioma detected by echocardiography, computed tomography, and magnetic resonance imaging. The multiple imaging modalities showed features which have been reported in cardiac hemangiomas. The tumor was surgically excised and the diagnosis of cardiac hemangioma was made. CONCLUSIONS: Cardiac hemangiomas are rare tumors with a variety of imaging features which may suggest the diagnosis. PMID- 19135391 TI - Thromboangiitis obliterans with multiple large vessel involvement: case report and analysis of immunophenotypes. AB - Thromboangiitis obliterans (TAO, Buerger's disease) is an idiopathic, recurrent, segmental, nonatherosclerotic, inflammatory, occlusive vascular disease with a poorly understood pathogenesis. Intestinal or multi-organ involvement is rare. Recent immunohistochemical analyses of ordinary TAO have indicated an inflammatory and immunologic pathogenesis. We report a case of TAO involving multiple large vessels. By immunohistochemistry, CD3+ T cells were revealed around the recanalization sites within the abdominal aorta. CD4+ T cells were almost equal in number to CD8+ T cells. These findings indicate the participation of inflammatory and immunologic processes in TAO with multi-organ involvement (as in ordinary TAO). PMID- 19135392 TI - The community use of rescue medication for prolonged epileptic seizures in children. AB - OBJECTIVES: The objectives for this study were to determine the prevalence of use, safety and efficacy of different preparations of rescue medication used for prolonged seizures in children in the community and to use the information gained to inform good practice within the network. METHODS: For all children in the EPIC area who had been prescribed epilepsy rescue medication for use in the community a questionnaire was completed by the clinician for each child relating to rescue medication prescribed, the epilepsy syndrome and seizure type. A questionnaire was also completed by the carers about their experience of the use of rescue medication in their child. RESULTS: A total of 203 paediatrician questionnaires were returned and 190 parent/carer questionnaires were returned. Buccal Midazolam was the most popular rescue medication (Buccal Midazolam 110, Rectal Diazepam 85, Paraldehyde 8). Over 80% of the children had a community care plan in place. 90% of carers recalled receiving instruction/training in the administration of rescue medication. The majority (73%) of carers perceived Buccal Midazolam and Rectal Diazepam to be effective in preventing hospital admission. CONCLUSION: Buccal Midazolam in its propriety form is the rescue medication most commonly prescribed by EPIC epilepsy paediatricians. It was felt by families to be the safest (least side effects reported) and the most effective (most likely to terminate seizures) of the rescue medications. Children with severe epilepsy benefit from the use of rescue medications and the number of hospital admissions for prolonged seizures is reduced. PMID- 19135394 TI - The influence of severe prolonged exercise restriction on the mechanical and structural properties of bone in an avian model. AB - Many studies have described the effects of exercise restriction on the mammalian skeleton. In particular, human and animal models have shown that reduction in weight bearing leads to generalised bone loss and deterioration of its mechanical properties. The aim of this study was to assess the effect of prolonged exercise restriction coupled with heavy calcium demands on the micro-structural, compositional and mechanical properties of the avian skeleton. The tibiae and humeri of 2-year-old laying hens housed in conventional caging (CC) and free range (FR) housing systems were compared by mechanical testing and micro-computed tomography (microCT) scanning. Analyses of cortical, cancellous and medullary bone were performed. Mechanical testing revealed that the tibiae and humeri of birds from the FR group had superior mechanical properties relative to those of the CC group, and microCT scanning indicated larger cortical and lower medullary regions in FR group bones. Cancellous bone analysis revealed higher trabecular thickness and a higher bone volume fraction in the FR group, but no difference in mineral density. The biomechanical superiority of bones from the FR group was primarily due to structural rather than compositional differences, and this was reflected in both the cortical and cancellous components of the bones. The study demonstrated that prolonged exercise restriction in laying hens resulted in major structural and mechanical effects on the bird skeleton. PMID- 19135393 TI - The constant region of the membrane immunoglobulin mediates B cell-receptor clustering and signaling in response to membrane antigens. AB - B cells are activated in vivo after the B cell receptors (BCRs) bind to antigens captured on the surfaces of antigen-presenting cells. Antigen binding results in BCR microclustering and signaling; however, the molecular nature of the signaling active BCR clusters is not well understood. Using single-molecule imaging techniques, we provide evidence that within microclusters, the binding of monovalent membrane antigens results in the assembly of immobile signaling-active BCR oligomers. The oligomerization depends on interactions between the membrane proximal Cmicro4 domains of the membrane immunoglobulin that are both necessary and sufficient for assembly. Antigen-bound BCRs that lacked the Cmicro4 domain failed to cluster and signal, and conversely, Cmicro4 domains alone clustered spontaneously and activated B cells. These results support a unique mechanism for the initiation of BCR signaling in which antigen binding induces a conformational change in the Fc portion of the BCR, revealing an interface that promotes BCR clustering. PMID- 19135396 TI - Theoretical analysis of solvent effects on nitrogen NMR chemical shifts in oxazoles and oxadiazoles. AB - Using quantum chemistry methods we have evaluated the solvent effects on the (14)N NMR chemical shifts in five oxa- and oxadiazoles dissolved in twelve solvents. These solvents differ in their polarity with the dielectric constants varying from 2 to 80. Moreover, three of them have a hydrogen-bond donor character. All possible hydrogen-bonding in the water solution with the oxygen and nitrogen (hydrogen-acceptor) centers in oxazoles (2) and oxadiazoles (3) have been considered in our studies. It has been shown that both the pure solvent and hydrogen-bonding effects are significant and result in (14)N magnetic shielding increase. In water solutions the pure solvent effect is larger than the hydrogen bonding effect. In addition, the solvent effect has been analyzed in terms of its direct and indirect contributions. It should be emphasized that our theoretical results for (14)N chemical shifts in oxa- and oxadiazoles remain in a very good agreement with the accurate experimental data. PMID- 19135395 TI - Molecular and cellular insights into a distinct myopathy of Great Dane dogs. AB - A myopathy in the Great Dane dog with characteristic pathological and molecular features is reported. Young adults present with progressive weakness and generalised muscle atrophy. To better define this condition, an investigation using histopathology, confocal microscopy, biochemistry and microarray analysis was undertaken. The skeletal muscles of affected dogs exhibited increased oxidative fibre phenotype and core fibre lesions characterised by the disruption of the sarcomeric architecture and the accumulation of mitochondrial organelles. Affected muscles displayed co-ordinated expression of genes consistent with a slow-oxidative phenotype, which was possibly a compensatory response to chronic muscle damage. There was disruption of Z-lines in affected muscles which, at the molecular level, manifested as transcriptional dysregulation of several Z-line associated genes, including alpha-actinin, myotilin, desmin, vimentin and telethonin. The pathology of this canine myopathy is distinct from that of human central core myopathies that are characterised by cores devoid of mitochondria and by the presence of myofibrillar breakdown products. PMID- 19135397 TI - Optimized sampling patterns for multidimensional T2 experiments. AB - Non-uniform sampling in multidimensional NMR shows great promise to significantly decrease experimental acquisition times, especially for relaxation experiments for which peak locations are already known. In this paper we present a method for optimizing the non-uniform sampling points such that the noise amplification and numerical instabilities are minimized. In particular, the minimum singular value of the Moore-Penrose pseudo-inverse is maximized using sequential semi-definite programming, thereby minimizing the worst-case errors. We test this method numerically on a set of assignment data from the proteins ubiquitin (in both folded and unfolded states) and RIalpha (119-244), a cAMP-binding regulatory subunit of protein kinase A (PKA). This test indicates that optimizing more than doubles the efficiency over random selection of points, and the efficiency increases as we go to higher dimensions. PMID- 19135398 TI - A neurotropic lymphoma in the brachial plexus of a cat. AB - A 7-year-old, intact male domestic shorthair cat was presented with a progressive, non-weight-bearing lameness of the right forelimb. A neurological examination of the patient at the Small Animal Clinic revealed a paralysis of the radial nerve with sustained cutaneous sensation and a hyperaesthetic response. Further clinical, radiological and pathological findings lead to a diagnosis of a primary, neurotropic B-cell lymphoma in the brachial plexus. PMID- 19135399 TI - Frequency of isolation of various subtypes and antimicrobial resistance of Shigella from urban slums of Karachi, Pakistan. AB - OBJECTIVES: Shigellosis remains a major public health problem in developing countries. Antimicrobial resistance has complicated the empirical treatment. Knowledge of serotypes is crucial in vaccine development, as cross-protection between various serotypes is limited. Therefore we conducted a prospective study to determine the frequency of isolation of Shigella serotypes and antimicrobial resistance. METHODS: Stool samples from 8155 individuals, collected through a surveillance study conducted in four slums of Karachi from January 2002 to March 2004, were cultured. RESULTS: Shigella was isolated in 394 (4.8%) of 8155 patients presenting with diarrhea. Two hundred and forty-two (62%) isolates were Shigella flexneri, 72 (18%) were Shigella sonnei, 43 (11%) were Shigella boydii, and 37 (9%) were Shigella dysenteriae. Thirteen S. flexneri serotypes were identified, of which the most frequent were 2a (38), 6 (37), and 1b (25), followed by 2b (23). Only 22 (5.6%) Shigella isolates were found to be pan susceptible. Large proportions of isolates were resistant to co-trimoxazole (89% S. flexneri, 81% S. dysenteriae, 80% S. sonnei, and 56% S. boydii) and ampicillin (87% S. flexneri, 68% S. dysenteriae, 35% S. boydii, and 4% S. sonnei). CONCLUSIONS: Concurrent circulation of multiple strains with high resistance is worrying and mandates surveillance at the national level to facilitate the control of shigellosis. PMID- 19135400 TI - High prevalence of beta-lactam-resistant Haemophilus influenzae type b isolates derived from respiratory tract specimens in Japanese patients. AB - OBJECTIVE: Serotypeable strains of Haemophilus influenzae, which can cause invasive infections, are found in the respiratory tract at low frequencies. We compared the antibiotic resistance of the typeable and nontypeable strains of H. influenzae in respiratory tract specimens obtained in Japan. METHODS: We determined the serotypes and the antibiotic susceptibilities of 440 clinical H. influenzae strains isolated from respiratory tract specimens. We also examined the prevalence of genotypes that are associated with beta-lactam resistance. RESULTS: The majority of the strains were nontypeable (421 strains, 95.7%). The remainder belonged to serotypes b (10 strains, 2.3%), e (three strains, 0.7%), or f (six strains, 1.4%). The type b strains exhibited the expression of beta lactamase and resistance mutations in penicillin-binding protein 3 with significantly higher frequencies than other strains. CONCLUSIONS: H. influenzae type b strains, which are associated with meningitis and bacteremia, derived from respiratory tract specimens, shared more beta-lactam-resistant mechanisms than nontypeable and other serotype strains. PMID- 19135401 TI - Peritoneal tuberculosis in pregnancy mimicking advanced ovarian cancer: a plea to avoid hasty, radical and irreversible surgical decisions. AB - Tuberculous peritonitis is rare in most Western counties, and can cause significant diagnostic and therapeutic problems. A 28-year-old pregnant female presented with nausea and vomiting, right lower quadrant abdominal pain, fever and intra-abdominal fluid. During surgery for presumed complicated acute appendicitis, many small masses (considered to be 'implants') were found within the peritoneal cavity, with a larger mass in the pelvis, mainly on the right. The clinical intra-operative diagnosis was advanced ovarian cancer and multiple biopsies were taken. The histological diagnosis was peritoneal tuberculosis. The patient was successfully treated conservatively. Hasty decisions to undertake radical and irreversible surgery should be avoided; this type of surgery should be performed only after histological confirmation. PMID- 19135402 TI - [Replacement and resurfacing prosthesis of the distal radius: a new therapeutic concept]. AB - Complex intra-articular distal radius fractures represent a challenging therapeutic problem. We propose in these difficult cases to replace and to resurface the distal radius by a prosthesis, especially in fractures related to aging and osteoporosis. We have designed an anatomic prosthesis to replace and resurface the distal radius. The design was obtained from radiographs, tomograms and anatomical specimens. The prosthesis is composed of a radial stem and an epiphyseal-metaphyseal bloc articulating both with the carpal condyle and the ulnar head. The epiphyseal-metaphyseal bloc can rotate 10 degrees on the longitudinal axis of the stem. From March 2005 to February 2007, six patients were operated on: five women and one man, the mean age was 73 years (38-87). Five cases were elderly and osteoporotic patients: four had complex intra-articular fractures and one had a malunion complicated by extensor tendon ruptures. In one case, the procedure was performed on a young man with a pathologic fracture (giant cell tumor). The patients were reviewed with a mean follow-up of 27 months (13-36 months). Five patients are satisfied or very satisfied but one patient is dissatisfied due to a stiff wrist after an algodystrophic syndrome. The mean mobilities are: extension 65 degrees, flexion 30 degrees, ulnar inclination 20 degrees, radial inclination 20 degrees, pronation 60 degrees, supination 50 degrees. The mean strength was 80% of the contralateral side. The implants are perfectly stable on the radiographs. Replacement and resurfacing of the distal radius by a prosthesis articulating both with the carpal condyle and ulnar head is possible. The preliminary clinical results are good. This prosthesis takes into account the transmission of the pronosupinator couples through the wrist. The longitudinal rotation between the stem and the epiphyseal-metaphyseal bloc has three goals: the first is to decrease the constraints born from the pronosupination torques, the second is to improve the congruence with the carpal condyle, the third is to increase radio-ulnocarpal stability. The treatment of complex intra-articular distal radius fractures by replacement and resurfacing prosthesis is a solution we can now consider in elderly patients with osteoporosis. This solution seems ideal for distal radius tumors. PMID- 19135403 TI - 3D nonrigid registration via optimal mass transport on the GPU. AB - In this paper, we present a new computationally efficient numerical scheme for the minimizing flow approach for optimal mass transport (OMT) with applications to non-rigid 3D image registration. The approach utilizes all of the gray-scale data in both images, and the optimal mapping from image A to image B is the inverse of the optimal mapping from B to A. Further, no landmarks need to be specified, and the minimizer of the distance functional involved is unique. Our implementation also employs multigrid, and parallel methodologies on a consumer graphics processing unit (GPU) for fast computation. Although computing the optimal map has been shown to be computationally expensive in the past, we show that our approach is orders of magnitude faster then previous work and is capable of finding transport maps with optimality measures (mean curl) previously unattainable by other works (which directly influences the accuracy of registration). We give results where the algorithm was used to compute non-rigid registrations of 3D synthetic data as well as intra-patient pre-operative and post-operative 3D brain MRI datasets. PMID- 19135404 TI - Novelty signals: a window into hippocampal information processing. AB - The function of the human hippocampus is a contentious subject among neuroscientists. Theoreticians have long viewed the hippocampus as a computational device, with researchers in humans increasingly adopting this perspective, buoyed by recent reports that its role is not limited to declarative memory. Here, we set out a new strategy for discovering the nature of information processing within the human hippocampus. We argue that novelty responses, measured by functional magnetic resonance imaging, provide a window into the neural representations and computations sustained by the hippocampus. More generally, we suggest that a renewed emphasis on the information processing qualities of the human hippocampus offers the promise of a long awaited union between theoretical and empirical research across species. PMID- 19135405 TI - Socioeconomic status and the developing brain. AB - Childhood socioeconomic status (SES) is associated with cognitive achievement throughout life. How does SES relate to brain development, and what are the mechanisms by which SES might exert its influence? We review studies in which behavioral, electrophysiological and neuroimaging methods have been used to characterize SES disparities in neurocognitive function. These studies indicate that SES is an important predictor of neurocognitive performance, particularly of language and executive function, and that SES differences are found in neural processing even when performance levels are equal. Implications for basic cognitive neuroscience and for understanding and ameliorating the problems related to childhood poverty are discussed. PMID- 19135406 TI - Looking for cognition in the structure within the noise. AB - Neural activity in the mammalian CNS is determined by both observable processes, such as sensory stimuli or motor output, and covert, internal cognitive processes that cannot be directly observed. We propose methods to identify these cognitive processes by examining the covert structure within the apparent 'noise' in spike trains. Contemporary analyses of neural codes include encoding (tuning curves derived from spike trains and behavioral, sensory or motor variables), decoding (reconstructing behavioral, sensory or motor variables from spike trains and hypothesized tuning curves) and generative models (predicting the spike trains from hypothesized encoding models and decoded variables). We review examples of each of these processes in hippocampal activity, and propose a general methodology to examine cognitive processes via the identification of dynamic changes in covert variables. PMID- 19135407 TI - Turning on the spotlight--using light to monitor and characterize bacterial effector secretion and translocation. AB - Secretion and translocation of bacterial pathogen effectors into host cells via dedicated secretion machineries like type III secretion systems (T3SSs) or type IV secretion systems (T4SSs) is a key feature employed by pathogens to attack host cells. Innovative fluorescence and imaging approaches have blossomed during recent years, and became instrumental in revealing the dynamics of effector secretion and function in interfering with host cellular processes, particularly signaling events, gene expression regulation, membrane trafficking, and autophagy. Furthermore, imaging-based screening approaches have demonstrated the mode of action of several bacterial effectors upon host cellular translocation. The rapid technological advancement of imaging technologies indicates that these techniques will continue to be at the center of numerous future breakthroughs delineating the dynamic processes of bacterial effector actions. PMID- 19135408 TI - Real-time live imaging to study bacterial infections in vivo. AB - In vitro studies have been essential to describe the molecular details of bacteria-host cell interactions in general and the functions of bacterial effector proteins in particular. Recent advancements in in vivo imaging techniques are facilitating the next logical step to visualize the dynamic infection process as it happens within the living host while analyzing the role of bacterial effector proteins in vivo. Data obtained from this emerging field of 'tissue microbiology', combined with the massive knowledge base generated from research in 'cellular microbiology' will eventually provide a complete picture of the complex infection process. PMID- 19135409 TI - Spectrophotometric study for the reaction between fluvoxamine and 1,2 naphthoquinone-4-sulphonate: Kinetic, mechanism and use for determination of fluvoxamine in its dosage forms. AB - Spectrophotometric study was carried out, for the first time, to investigate the reaction between the antidepressant fluvoxamine (FXM) and 1,2-naphthoquinone-4 sulphonate (NQS) reagent. In alkaline medium (pH 9), an orange-colored product exhibiting maximum absorption peak (lambda(max)) at 470nm was produced. The kinetics of the reaction was investigated and its activation energy was found to be 2.65kcalmol(-1). Because of this low activation energy, the reaction proceeded easily. The stoichiometry of the reaction was determined and the reaction mechanism was postulated. This color-developing reaction was successfully employed in the development of simple and rapid spectrophotometric method for determination of FXM in its pharmaceutical dosage forms. Under the optimized reaction conditions, Beer's law correlating the absorbance (A) with FXM concentration (C) was obeyed in the range of 0.6-8microgml(-1). The regression equation for the calibration data was A=0.0086+0.1348C, with good correlation coefficient (0.9996). The molar absorptivity (epsilon) was 5.9x10(4)lmol(-1)cm( 1). The limits of detection and quantification were 0.2 and 0.6microgml(-1), respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 2%. The proposed method was successfully applied to the determination of FXM in its pharmaceutical tablets with good accuracy and precisions; the label claim percentage was 100.47+/-0.96%. The results obtained by the proposed method were comparable with those obtained by the official method. The proposed method is superior to all the previously reported spectrophotometric methods for determination of FXM in terms of its simplicity and sensitivity. The method is practical and valuable for its routine application in quality control laboratories for analysis of FXM. PMID- 19135410 TI - Direct identification of human enterovirus serotypes in cerebrospinal fluid by amplification and sequencing of the VP1 region. AB - BACKGROUND: Human enteroviruses (HEV) are a major cause of meningitis and other neurological disease. Identification of HEV serotypes in clinical cases is important for monitoring emergence of more pathogenic variants, epidemiological surveillance and investigating sources of infection. Serotype identification is currently problematic following the widespread adoption of polymerase chain reaction (PCR)-based methods for HEV detection in place of virus culture. OBJECTIVES: To develop a reliable, sensitive method to identify species A and B serotypes directly from cerebrospinal fluid (CSF) specimens. STUDY DESIGN: A nested-PCR was used to amplify VP1 region sequences of HEV species A and B, that enabled unambiguous serotype identification by comparison with reference strains. RESULTS: 62 from 64 diagnostic CSF samples collected over a 19-month study period were successfully amplified (97% sensitivity), compared with 9/22 (41%) identified by virus culture of co-referred faecal and throat swab samples. Among these, 60 samples contained species B and 2 samples contained species A serotypes (coxsackievirus A6 and enterovirus 71) were identified. Rapid changes in serotype frequencies and diversity were observed; echovirus (E) type 9 infections predominated in early 2007, to be replaced by E30 later in the year and followed by a diverse range of eight different species B serotypes in 2008. CONCLUSIONS: The availability of a simple and rapid method for identification of serotypes and individual HEV strains or clusters directly from CSF will be of substantial value in surveillance, understanding more about serotype-associated differences in disease and monitoring the global spread of pathogenic variants such as enterovirus 71. PMID- 19135411 TI - Changes in long term neural connectivity following psychological trauma. AB - OBJECTIVE: Neural connectivity differences between adults reporting childhood, adulthood or no history of trauma were examined. METHODS: A total of 39 participants completed the Post-traumatic Stress Diagnostic Scale (PDS; Foa EB. Post-traumatic Stress Diagnostic Scale (PDS) Manual. Minneapolis, MN: National Computer Systems, 1995), a Word Memory Task (WMT; [McNally RJ, Metzger LJ, Lasko NB, Clancy SA, Pitman RK. Directed forgetting of trauma cues in adult survivors of childhood sexual abuse with and without post-traumatic stress disorder. J Abnorm Psychol 1998;107:596-601]) and EEG analysis. Intelligence was not assessed during the study. RESULTS: As predicted, those with childhood trauma had significantly higher EEG coherence than those with either adulthood trauma or no past trauma. CONCLUSIONS: Significant differences were observed over frontal, central, temporal and parietal areas. Evidence was found suggesting that childhood psychological trauma may have a lasting impact on neuronal connectivity. SIGNIFICANCE: This is the first study to demonstrate the suspected long term effect of trauma over central, temporal and parietal areas. Long term neural correlates of childhood and adult trauma appear to suggest information processing differences--differences that may, eventually, lead to better interventions following trauma. PMID- 19135412 TI - Reliability of transcranial magnetic stimulation-related measurements of tibialis anterior muscle in healthy subjects. AB - OBJECTIVE: The main purpose of this study was to determine the reliability of transcranial magnetic stimulation (TMS)-related measurements linked to the corticospinal control of the tibialis anterior (TA) muscle in healthy subjects. METHODS: Inter-investigator and intra-investigator (within-session with a 1.5-h interval; between-session with a 4-week interval) reliability of the motor threshold, recruitment curve with its slope and MEP area at the plateau, MEP latency, maximum MEP (MEPmax) area, and duration of the maximum and minimum silent period (SPmax and SPmin) were assessed in 50 (29 men and 21 women) healthy subjects (mean age 44.8 years, range 22-74 years) using intraclass correlation coefficients (ICC), standard error of measurement (SEM) and smallest real difference (SRD). RESULTS: The high ICC values and lower SEM and SRD values indicate a good intra- and inter-investigator reliability for motor threshold (ICC range 0.94-0.98), MEP latency (ICC range 0.79-0.93), SPmax (ICC range 0.89 0.95) and SPmin (ICC range 0.79-0.81) in healthy subjects. CONCLUSIONS: These results suggest that the TMS-related measurements investigated are reliable in healthy subjects. SIGNIFICANCE: These data could be useful in further studies on cortical excitability changes, such as those induced by therapeutic interventions (e.g. rehabilitative treatment). PMID- 19135413 TI - Pulmonary capillary wedge pressure and pulmonary arterial pressure in heart failure patients with sleep-disordered breathing. AB - BACKGROUND: There is a high prevalence of central sleep apnea (CSA) in patients with chronic heart failure (CHF). The present study investigates the hypotheses that CSA in CHF patients reflects heart failure severity as measured by cardiac index (CI), pulmonary artery pressure (PAP) and pulmonary capillary wedge pressure (PCWP). METHODS: In 105 patients with stable CHF (NYHAII, LV-EF40%) cardiorespiratory polygraphy and simultaneous right and left heart catheterization was performed. RESULTS: CSA was present in 58% and obstructive sleep apnea (OSA) in 23% of patients. In CSA patients, PAP and PCWP were significantly higher when compared to patients without SDB. In CSA patients, but not in OSA patients, PCWP showed a significant correlation with apnea-hypopnea index (AHI; r=0.41, p=0.005), apnea index (AI; r=0.44, p=0.003) and central AI (cAI; r=0.358, p=0.015). Cardiac index was more impaired in CSA (1.93+/-0.5 l/min/m(2)) than in OSA patients (2.55+/-1.0 l/min/m(2)) or those without SDB (2.22+/-0.4 l/min/m(2)). A negative correlation of CI and cAI (r=-0.344, p=0.008), AI (r=-0.31, p=0.02) and AHI (r=-0.21, p<0.05) was documented exclusively in CSA patients. CONCLUSION: The present study supports the hypotheses that the occurrence and severity of CSA in CHF patients reflects heart failure severity. PMID- 19135414 TI - Lead--still a health threat for marksmen. AB - OBJECT: Indoor shooting is a popular recreational activity all over the world. It is also a known source of exposure to lead fumes and dusts for the general, i.e. the non-occupationally exposed population. This study aimed at reporting the lead levels in blood of indoor shooters in Germany in association with personal behaviour at shooting ranges. METHODS: Seven marksmen were examined via self administered questionnaires. Lead level in blood (BLL) as well as airborne lead concentration (personal air sampling (PAS)) were detected. RESULTS: All measured blood lead levels (median: 290 microglead/L; range: 240-450 microglead/L) exceeded the values of the German general population (50 microg/L). Shooters sweeping the range after training seemed especially exposed (PAS: 7.14 mglead/m3). CONCLUSION: While extensive public effort is undertaken to prevent the general population from environmental lead exposure this short report gives reason to keep recreational shooting in mind when facing elevated blood lead levels or clinical symptoms possibly associated with lead intoxication. PMID- 19135415 TI - Palonosetron plus dexamethasone versus granisetron plus dexamethasone for prevention of nausea and vomiting during chemotherapy: a double-blind, double dummy, randomised, comparative phase III trial. AB - BACKGROUND: Palonosetron is a second-generation 5-hydroxytryptamine 3 (5-HT(3)) receptor antagonist that has shown better efficacy than ondansetron and dolasetron in preventing chemotherapy-induced nausea and vomiting (CINV) in patients receiving moderately emetogenic chemotherapy, and similar efficacy to ondansetron in preventing CINV in patients receiving highly emetogenic chemotherapy. In this phase III, multicentre, randomised, double-blind, double dummy, stratified, parallel-group, active-comparator trial, we assessed the efficacy and safety of palonosetron versus granisetron for chemotherapy-induced nausea and vomiting, both of which were administered with dexamethasone in patients receiving highly emetogenic chemotherapy. METHODS: Between July 5, 2006, and May 31, 2007, 1143 patients with cancer who were receiving highly emetogenic chemotherapy (ie, cisplatin, or an anthracycline and cyclophosphamide combination [AC/EC]) were recruited from 75 institutions in Japan, and randomly assigned to either single-dose palonosetron (0.75 mg), or granisetron (40 microg/kg) 30 min before chemotherapy on day 1, both with dexamethasone (16 mg intravenously) on day 1 followed by additional doses (8 mg intravenously for patients receiving cisplatin or 4 mg orally for patients receiving AC/EC) on days 2 and 3. A non deterministic minimisation method with a stochastic-biased coin was applied to the randomisation of patients. Covariates known to effect emetic risk, such as sex, age, and type of highly emetogenic chemotherapy, were used as stratification factors of minimisation to ensure balance between the treatment groups. Primary endpoints were the proportion of patients with a complete response (defined as no emetic episodes and no rescue medication) during the acute phase (0-24 h postchemotherapy; non-inferiority comparison with granisetron) and the proportion of patients with a complete response during the delayed phase (24-120 h postchemotherapy; superiority comparison with granisetron). The non-inferiority margin was predefined in the study protocol as a 10% difference between groups in the proportion of patients with complete response. The palonosetron dose of 0.75 mg was chosen on the basis of two dose-determining trials in Japanese patients. All patients who received study treatment and highly emetogenic chemotherapy were included in the efficacy analyses (modified intention to treat). This trial is registered with ClinicalTrials.gov, number NCT00359567. FINDINGS: 1114 patients were included in the efficacy analyses: 555 patients in the palonosetron group and 559 patients in the granisetron group. 418 of 555 patients (75.3%) in the palonosetron group had complete response during the acute phase compared with 410 of 559 patients (73.3%) in the granisetron group (mean difference 2.9% [95% CI 2.70 to 7.27]). During the delayed phase, 315 of 555 patients (56.8%) had complete response in the palonosetron group compared with 249 of 559 patients (44.5%) in the granisetron group (p<0.0001). The main treatment-related adverse events were constipation (97 of 557 patients [17.4%] in the palonosetron group vs 88 of 562 [15.7%] in the granisetron group) and raised concentrations of serum aminotransferases (aspartate aminotransferase: 24 of 557 [4.3%] vs 34 of 562 [6.0%]; alanine aminotransferase: 16 of 557 [2.9%] vs 33 of 562 [5.9%]); no grade 4 main treatment-related adverse events were reported. INTERPRETATION: When administered with dexamethasone before highly emetogenic chemotherapy, palonosetron exerts efficacy against chemotherapy-induced nausea and vomiting which is non-inferior to that of granisetron in the acute phase and better than that of granisetron in the delayed phase, with a comparable safety profile for the two treatments. FUNDING: Taiho Pharmaceutical (Tokyo, Japan). PMID- 19135416 TI - Live immunization against East Coast fever--current status. AB - The infection-and-treatment method (ITM) for immunization of cattle against East Coast fever has historically been used only on a limited scale because of logistical and policy constraints. Recent large-scale deployment among pastoralists in Tanzania has stimulated demand. Concurrently, a suite of molecular tools, developed from the Theileria parva genome, has enabled improved quality control of the immunizing stabilate and post-immunization monitoring of the efficacy and biological impact of ITM in the field. This article outlines the current status of ITM immunization in the field, with associated developments in the molecular epidemiology of T. parva. PMID- 19135417 TI - Variation in Giardia: towards a taxonomic revision of the genus. AB - Taxonomic uncertainty has had a negative impact on our understanding of the epidemiology of Giardia infections, particularly the role of wild and domestic animals as sources of human infection. The lack of morphological criteria for species identification and the failure of cross-infection experiments to unequivocally determine host specificity have largely contributed to this uncertainty. However, over the past ten years, it has been possible not only to demonstrate extensive genetic heterogeneity among Giardia isolates from mammals but also to confirm levels of host specificity that were recognized by early taxonomists when they proposed a series of host-related species that we consider should now be re-established. PMID- 19135418 TI - HIV Gag p24 specific responses secreting IFN-gamma and/or IL-2 in treatment-naive individuals in acute infection early disease (AIED) are associated with low viral load. AB - HIV-specific immune responses in acute infection early disease (AIED) may be effective at controlling viral replication and in establishing viral load (VL) set point. However, evidence correlating the function and specificity of these responses with the VL set point is lacking. To address this issue, we screened cells from 59 treatment-naive HIV infected individuals (33 in AIED and 26 progressors) for responses to the entire HIV proteome using a dual color ELISPOT assay detecting 3 functional lymphocyte populations: single IFN-gamma, dual IFN gamma/IL-2 and single IL-2 secreting cells. Responses characterized by dual secreting cells contributed more to the HIV specific response in AIED versus chronic infection. Of responses directed to individual HIV gene products the magnitude and breadth of only Gag p24-specific responses for the 3 functional subsets were associated with lower concurrent or set point VL. Therefore the early appearance of broader and more intense Gag-p24-specific responses may be a determinant of subsequent VL. PMID- 19135419 TI - Anti-Siglec-F antibody inhibits oral egg allergen induced intestinal eosinophilic inflammation in a mouse model. AB - Siglec-F is a sialic acid binding immunoglobulin-superfamily receptor that is highly expressed on eosinophils. We have used a mouse model of oral egg ovalbumin (OVA)-induced eosinophilic inflammation of the gastro-intestinal mucosa associated with diarrhea and weight loss to determine whether administering an anti-Siglec-F antibody would reduce levels of intestinal mucosal eosinophilic inflammation. Mice administered the anti-Siglec-F antibody had significantly lower levels of intestinal eosinophilic inflammation, and this was associated with reduced intestinal permeability changes, normalization of intestinal villous crypt height, and restoration of weight gain. The reduced numbers of intestinal eosinophils in anti-Siglec-F antibody treated mice was associated with significantly reduced numbers of bone marrow and peripheral blood eosinophils, but was not associated with significant changes in the numbers of proliferating or apoptotic jejunal eosinophils. In addition, the anti-Siglec-F Ab reduced Th2 cytokines and IgE levels. Overall, these studies demonstrate that administration of an anti-Siglec-F antibody significantly reduces levels of eosinophilic inflammation in the intestinal mucosa and that this was associated with reduced intestinal permeability changes, normalization of intestinal villous crypt height, and restoration of weight gain. PMID- 19135420 TI - Superhydrophobic effect on the adsorption of human serum albumin. AB - Analytical protocol greatly influences the measurement of human serum albumin (HSA) adsorption to commercial expanded polytetrafluororethylene (ePTFE) exhibiting superhydrophobic wetting properties. Degassing of buffer solutions and evacuation of ePTFE adsorbent to remove trapped air immediately prior to contact with protein solutions are shown to be essential. Results obtained with ePTFE as a prototypical superhydrophobic test material suggest that vacuum degassing should be applied in the measurement of protein adsorption to any surface exhibiting superhydrophobicity. Solution depletion quantified using radiometry ((125)I-labeled HSA) or electrophoresis yield different measures of adsorption, with nearly 4-fold higher surface concentrations of unlabeled HSA measured by the electrophoresis method. This outcome is attributed to the influence of the radiolabel on HSA hydrophilicity which decreases radiolabeled-HSA affinity for a hydrophobic adsorbent in comparison to unlabeled HSA. These results indicate that radiometry underestimates the actual amount of protein adsorbed to a particular material. Removal of radiolabeled HSA adsorbed to ePTFE by 3x serial buffer rinses also shows that the remaining "bound fraction" was about 35% lower than the amount measured by radiometric depletion. This observation implies that measurement of protein bound after surface rinsing significantly underestimates the actual amount of protein concentrated by adsorption into the surface region of a protein-contacting material. PMID- 19135421 TI - Altered structural and mechanical properties in decellularized rabbit carotid arteries. AB - Recently, major achievements in creating decellularized whole tissue scaffolds have drawn considerable attention to decellularization as a promising approach for tissue engineering. Decellularized tissues are expected to have mechanical strength and structure similar to the native tissues from which they are derived. However, numerous studies have shown that mechanical properties change after decellularization. Often, tissue structure is observed by histology and electron microscopy, but the structural alterations that may have occurred are not always evident. Here, a variety of techniques were used to investigate changes in tissue structure and relate them to altered mechanical behavior in decellularized rabbit carotid arteries. Histology and scanning electron microscopy revealed that major extracellular matrix components were preserved and fibers appeared intact, although collagen appeared looser and less crimped after decellularization. Transmission electron microscopy confirmed the presence of proteoglycans (PG), but there was decreased PG density and increased spacing between collagen fibrils. Mechanical testing and opening angle measurements showed that decellularized arteries had significantly increased stiffness, decreased extensibility and decreased residual stress compared with native arteries. Small angle light scattering revealed that fibers had increased mobility and that structural integrity was compromised in decellularized arteries. Taken together, these studies revealed structural alterations that could be related to changes in mechanical properties. Further studies are warranted to determine the specific effects of different decellularization methods on the structure and performance of decellularized arteries used as vascular grafts. PMID- 19135422 TI - Strong bonding strength between HA and (NH4)2S2O8-treated carbon/carbon composite by hydrothermal treatment and induction heating. AB - Carbon/carbon composite with hydroxyapatite (HA) coating is an attractive material in the dental and orthopedic fields, but the reported bonding strength between them was very poor. In this study, a compact crystalline HA coating on (NH(4))(2)S(2)O(8)-treated C/C substrate about 10 microm in width was obtained by hydrothermal treatment and induction heating. The microstructure, composition and morphologies of the as-prepared coatings were identified by X-ray diffraction, scanning electron microscopy and energy dispersive spectroscopy. A strong shear strength averaging 74.2 MPa between C/C substrate and HA was achieved and adhesion failures were observed more frequently than cohesion failures. The coating adhesion measured using a scratch test was 23 N and the reasons for this are discussed. PMID- 19135423 TI - Solubility of strontium-substituted apatite by solid titration. AB - Solid titration was used to explore the solubility isotherms of partially (Srx HAp, x=1, 5, 10, 40, 60 mol.%) and fully substituted strontium hydroxyapatite (Sr HAp). Solubility increased with increasing strontium content. No phase other than strontium-substituted HAp, corresponding to the original titrant, was detected in the solid present at equilibrium; in particular, dicalcium hydrogen phosphate was not detected at low pH. The increase in solubility with strontium content is interpreted as a destabilization of the crystal structure by the larger strontium ion. Carbonated HAp was formed in simulated body fluid containing carbonate on seeding with Sr10-HAp, but the precipitate was strontium-substituted on seeding with Sr-HAp. Strontium-substituted HAp might be usable as a template for the growth of new bone, since nucleation appears to be facilitated. PMID- 19135424 TI - Mass spectrometric quantification of neutral and sialylated N-glycans from a recombinant therapeutic glycoprotein produced in the two Chinese hamster ovary cell lines. AB - Quality control and assurance of glycan profiles of a recombinant glycoprotein from lot to lot is a critical issue in the pharmaceutical industry. To develop an easy and simple quantitative and qualitative glycan profile method based on matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS), the modification with Girard's reagent T (GT) was exploited. Because GT-derivatized quantification of oligosaccharides using MALDI-TOF MS is possible only with neutral glycans, sialylated glycans are not subjected to quantitative analysis with MALDI-TOF MS. To solve this problem, mild methyl esterification and subsequent GT derivatization were employed, enabling us to perform rapid qualitative and quantitative analysis of sialylated and neutral N linked oligosaccharides using MALDI-TOF MS. This modified method was used in the comparative quantification of N-glycans from the recombinant therapeutic glycoprotein expressed in two different Chinese hamster ovary (CHO) cell lines. The percentages of sialylated N-glycans to total were 22.5 and 5.2% in CHO-I and CHO-II cells, respectively, resulting in a significant difference in the biological activity of the recombinant glycoprotein. PMID- 19135425 TI - Effect of Wnt-1 inducible signaling pathway protein-2 (WISP-2/CCN5), a downstream protein of Wnt signaling, on adipocyte differentiation. AB - Wnt signaling negatively regulates adipocyte differentiation, and ectopic expression of Wnt-1 in 3T3-L1 cells induces several downstream molecules of Wnt signaling, including Wnt-1 inducible signaling pathway protein (WISP)-2. In this study, we examined the role of WISP-2 in the process of adipocyte differentiation using an in vitro cell culture system. In the differentiation of 3T3-L1 cells, WISP-2 expression was observed in growing cells and declined thereafter. In the mitotic clonal expansion phase of adipocyte differentiation, WISP-2 expression was transiently down-regulated concurrently with up-regulation of CCAAT/enhancer binding protein delta expression. Treatment of 3T3-L1 cells in the differentiation medium with lithium, an activator of Wnt signaling, inhibited the differentiation process with concomitant induction of WISP-2. Treatment of differentiated cells with lithium induced de-differentiation as evidenced by profound reduction of peroxisome proliferator-activator receptor gamma expression and concomitant induction of WISP-2. However, de-differentiation of differentiated cells induced by tumor necrosis factor-alpha did not induce WISP-2 expression. To directly examine the effect of WISP-2 on adipocyte differentiation, 3T3-L1 cells were infected with a retrovirus carrying WISP-2. Although forced expression of WISP-2 inhibited preadipocyte proliferation, it had no effect on adipocyte differentiation. Thus, although WISP-2 is a downstream protein of Wnt signaling, the role of WISP-2 on adipocyte differentiation may be marginal, at least in this in vitro culture model. PMID- 19135426 TI - PPARgamma agonist pioglitazone reduces matrix metalloproteinase-9 activity and neuronal damage after focal cerebral ischemia. AB - Pioglitazone, a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, has shown protective effects against ischemic insult in various tissues. Pioglitazone is also reported to reduce matrix metalloproteinase (MMP) activity. MMPs can remodel extracellular matrix components in many pathological conditions. The current study was designed to investigate whether the neuroprotection of pioglitazone is related to its MMP inhibition in focal cerebral ischemia. Mice were subjected to 90 min focal ischemia and reperfusion. In gel zymography, pioglitazone reduced the upregulation of active form of MMP-9 after ischemia. In in situ zymograms, pioglitazone also reduced the gelatinase activity induced by ischemia. After co-incubation with pioglitazone, in situ gelatinase activity was directly reduced. Pioglitazone reduced the infarct volume significantly compared with controls. These results demonstrate that pioglitazone may reduce MMP-9 activity and neuronal damage following focal ischemia. The reduction of MMP-9 activity may have a possible therapeutic effect for the management of brain injury after focal ischemia. PMID- 19135427 TI - USP14 inhibits ER-associated degradation via interaction with IRE1alpha. AB - Accumulation of unfolded proteins within the endoplasmic reticulum (ER) lumen induces ER stress. Eukaryotic cells possess the ER quality control systems, the unfolded protein response (UPR), to adapt to ER stress. IRE1alpha is one of the ER stress receptors and mediates the UPR. Here, we identified ubiquitin specific protease (USP) 14 as a binding partner of IRE1alpha. USP14 interacted with the cytoplasmic region of IRE1alpha, and the endogenous interaction between USP14 and IRE1alpha was inhibited by ER stress. Overexpression of USP14 inhibited the ER associated degradation (ERAD) pathway, and USP14 depletion by small interfering RNA effectively activated ERAD. These findings suggest that USP14 is a novel player in the UPR by serving as a physiological inhibitor of ERAD under the non stressed condition. PMID- 19135429 TI - Environmental enrichment alters neuronal processing in the nucleus accumbens core during appetitive conditioning. AB - Although the core region of the nucleus accumbens (NAcc) has been implicated in motor control and the acquisition of appetitive learning, these processes are altered by environmental experience. To assess how environment influences neuronal processing in NAcc core, we recorded single-unit activity during acquisition of an appetitive learning task in which rats reared in an environmentally enriched condition (EC) learned the operant response (nosepoke into a lit hole) for sucrose reinforcement faster than rats reared in an isolated condition (IC). In the first training session, even before the emergence of learning differences, core neurons were more likely to respond (increase or decrease activity) during the operant and consummatory responses in EC than IC rats. By the third training session, when learning differences emerged, EC neurons continued to be more responsive than IC neurons, but in very different ways: the response shifted to the cues that signaled trial onset (1900 Hz tone and green LED) and reward availability (4500 Hz tone and yellow LED). Cue-related responding, moreover, was dominated by neuronal excitations. In contrast, post acquisition recordings revealed no EC-IC differences. Collectively, these results suggest that core neurons are initially more responsive to discrete, goal directed movements in EC rats, but as learning materializes, the neuronal response shifts to the cues that predict these movements. Thus, environmental experience alters core neuronal processing of both motor- and sensory-related events but at different stages over the course of learning. PMID- 19135428 TI - Paraquat is excluded by the blood brain barrier in rhesus macaque: An in vivo pet study. AB - Environmental factors have long been thought to have a role in the etiology of idiopathic Parkinson's disease (PD). Since the discovery of the selective neurotoxicity of MPTP to dopamine cells, suspicion has focused on paraquat, a common herbicide with chemical structure similar to 1-methyl-4-phenylpyridinium (MPP+), the MPTP metabolite responsible for its neurotoxicity. Although in vitro evidence for paraquat neurotoxicity to dopamine cells is well established, its in vivo effects have been ambiguous because paraquat is di-cationic in plasma, which raises questions about its ability to cross the blood brain barrier. This study assessed the brain uptake of [(11)C]-paraquat in adult male rhesus macaques using quantitative PET imaging. Results showed minimal uptake of [(11)C]-paraquat in the macaque brain. The highest concentrations of paraquat were seen in the pineal gland and the lateral ventricles. Global brain concentrations including those in known dopamine areas were consistent with the blood volume in those structures. This acute exposure study found that paraquat is excluded from the brain by the blood brain barrier and thus does not readily support the causative role of paraquat exposure in idiopathic Parkinson's disease. PMID- 19135430 TI - Iron is a potential key mediator of glutamate excitotoxicity in spinal cord motor neurons. AB - Threohydroxyaspartate (THA)-induced glutamate excitotoxicity in organotypic culture of rat spinal cord is a well-known model of motor neuron degeneration. THA causes accumulation of synaptic glutamate and over stimulation of the postsynaptic receptor by inhibiting glutamate uptake. This model has also been used to identify agents that inhibit glutamate excitotoxicity by increasing the expression of glutamate transporter. We now show that THA also increases iron level in rat spinal cord tissue, with concomitant modulation of key iron transport and storage proteins, including transferrin receptor, divalent metal ion transporter 1 and ferritin. More significantly, iron chelator deferoxamine (DFO) was able to completely prevent THA-induced motor neuron degeneration. The protective effect of DFO did not involve enhancing glutamate uptake. These data provide new mechanistic insight into THA-induced glutamate excitotoxicity and suggest that blocking THA-induced iron rise alone may be sufficient for prevention of glutamate excitotoxicity. PMID- 19135431 TI - Effects of environmental enrichment and physical activity on neurogenesis in transgenic PS1/APP mice. AB - Rodents exposed to environmental enrichment show many differences, including improved cognitive performance, when compared to those living in standard (impoverished) housing. The purpose of the present study was to determine if a selective increase in neurogenesis occurred in cognitively-protected Tg mice raised in an enriched environment compared to those reared in physical activity housing. At weaning, double Tg APP+PS1 mice were placed into one of three environments: complete environmental enrichment (CE), enhanced physical activity (PA), or individual, impoverished housing (IMP). At 9-10 months of age, Tg mice were injected with BrdU (100 mg/kg BID) followed by euthanasia either 24 h or 2 weeks after the last injection. Unbiased estimates of BrdU positive cells in the hippocampal subgranular zone revealed a significant increase in cellular proliferation in Tg mice raised in CE or PA compared to Tg mice reared in IMP housing. However, counts of BrdU birth-dated cells 2 weeks after labeling showed no difference among the three groups, indicating decreased survival of cells in those groups (CE and PA) with higher cellular proliferation rates in the neurogenic niche. Counts of calretinin-expressing cells, a marker of immature neurons, also indicated no difference among the three groups of mice. In view of our prior study showing that enhanced cognitive activity (but not enhanced physical activity) protects Tg mice against cognitive impairment, the present results indicate that increased generation and survival of new neurons in the hippocampal dentate gyrus is not involved with the cognitively-protective effects of complete CE in Alzheimer's transgenic mice. PMID- 19135432 TI - Enhancement of iron-catalyzed lipid peroxidation by acidosis in brain homogenate: comparative effect of diphenyl diselenide and ebselen. AB - Iron is more soluble at lower pH values; therefore we hypothesized that decreasing the environmental pH would lead to increased iron-mediated lipid peroxidation. Diphenyl diselenide and ebselen are potential candidates as neuroprotective agent, particularly in situations involving overproduction of free radicals and involving cellular pH fall. The aim of the present study was (a) to investigate the relationship between lipid peroxidation and acidosis in brain homogenate and (b) to test the influence of pH on the antioxidant properties of diphenyl diselenide and ebselen. For the purpose rat brain homogenate was incubated at different pH ranging from physiological to acidic values and extent of lipid peroxidation was measured. Thiobarbituric acid reactive species (TBARS) production significantly increased when homogenate was incubated in the pH (5.4-6.8) medium both in the absence and presence of Fe (II) as compared with physiological pH (7.4). These data indicate that lipid peroxidation processes, mediated by iron, are enhanced with decreasing extracellular pH. The iron mobilized may come from reserves where it is weakly bound. Diphenyl diselenide significantly protected TBARS production at all studied pH values while ebselen offered only a small statistically non significant protection. However, calculated IC(50) for TBARS inhibition indicated that pH did not change anti-oxidant activities of the tested compounds. This study provides in-vitro evidence for acidosis induced oxidative stress in brain homogenate and anti-oxidant action of diphenyl diselenide. PMID- 19135433 TI - Cumene hydroperoxide debilitates macrophage physiology by inducing oxidative stress: possible protection by alpha-tocopherol. AB - Macrophages, the major phagocytes of body, are largely dependent on membrane for their apposite functioning. Cum-OOH, a catalyst used in chemical and pharmaceutical industry, is a peroxidative agent, which may induce oxidative stress in macrophages hampering the integrity of their membrane. Alpha-tocopherol is known to protect the membrane from oxidative modulation and preserve its integrity. In the present study, we investigated the effect of Cum-OOH on physiology of macrophages and evaluated the protective effect of alpha-tocopherol against Cum-OOH-induced functional impairment. An in vitro exposure to 10-200 microM Cum-OOH altered redox balance of murine peritoneal macrophages and led to a severe physiological impairment. It markedly augmented the release of proinflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta and prostaglandin E(2)), lipopolysaccharide primed nitric oxide release and inducible nitric oxide synthase expression, and lysosomal hydrolases secretion. It mitigated respiratory burst and phagocytosis and intracellular killing of yeast (Saccharomyces cerevisiae). Mannose receptor, a major macrophage phagocytic receptor (also implicated in S. cerevisiae phagocytosis), exhibited a hampered recycling with its number being reduced to about 54% of the untreated, control cells following Cum-OOH exposure. A 24-h pretreatment of macrophages with 25 microM alpha-tocopherol preserved most of the assessed functions close to their corresponding control values. These data suggest that exposure to Cum-OOH may impair the physiology of immune cells such as macrophages and that supplementation with alpha-tocopherol can safeguard these cells against Cum-OOH toxicity. PMID- 19135434 TI - GastroPanel: evaluation of the usefulness in the diagnosis of gastro-duodenal mucosal alterations in children. AB - BACKGROUND: The combined evaluation of serum pepsinogens A (PGA) and C (PGC), gastrin-17 (G17) and anti-H. pylori antibodies (anti-H. pylori)(GastroPanel) has recently been proposed as a useful aid for investigating H. pylori-associated gastric mucosal inflammation. Our aim was to evaluate whether GastroPanel can correctly classify children who need or not endoscopy (EGD). METHODS: GastroPanel was performed in 554 consecutive children subjected to EGD. RESULTS: PGC and anti H. pylori were sensitive (82.5% and 73.1%) and specific (58.1% and 84.0%) indices of H. pylori infection. Antral H. pylori colonization density, inflammation and activity grades were correlated with PGC. PGC and G17 were significantly higher in children with celiac disease (14.9+/-0.88 microg/L and 5.6+/-0.79 pmol/L) than in controls (8.5+/-0.38 microg/L and 2.4+/-0.24 pmol/L). The best cut-offs to distinguish H. pylori infected children from controls were 7.45 microg/L for PGC, 4.2 pmol/L for G17, 18 U for anti-H. pylori and 25 microg/L for PGA. With these cut-offs, GastroPanel had a NPV of 89.6% and a PPV of 66.8%. CONCLUSIONS: A negative GastroPanel result in children with upper abdominal non alarm symptoms, should allow the paediatrician to reasonably rule out the presence of major gastro-duodenal diseases and therefore avoid EGD. PMID- 19135435 TI - Important roles of glycosylphosphatidylinositol (GPI)-specific phospholipase D and some GPI-anchored proteins in the pathogenesis of hepatocellular carcinoma. AB - OBJECTIVE: To investigate the roles of glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) in the pathogenesis of hepatocellular carcinoma (HCC). METHODS: The expression of the GPI-PLD in HCC was determined. The GPI-PLD gene was stably transfected in HepG2 cells and the proliferation of these cells was detected; CD55, CD59 and apoptotic cells were also analyzed. RESULTS: The serum GPI-PLD activities, the protein and mRNA levels of GPI-PLD in HCC patients were decreased by 40%, 60% and 56%, respectively. The killing rate of CDC against the positive clone cells was significantly increased, but the proliferative capacity was obviously decreased. The apoptotic rate in positive clones was increased. CONCLUSION: The expression of GPI-PLD decreases in HCC patients. The over expression of GPI-PLD in HepG2 cells increases their sensitivity to CDC killing, impairs proliferative capacity and promotes apoptosis. PMID- 19135436 TI - The role of Xenopus Rx-L in photoreceptor cell determination. AB - Members of the Rx (retinal homeobox) gene family play vital roles during eye development. In Xenopus, as in most other vertebrates, two Rx-type genes have been described. While Rx1 deficiency led to loss of optic vesicles and impaired the proper development of ventral forebrain structures, a recently isolated second Rx-gene, Rx-L, seems to function in late retinogenesis. Here, we report that the specific suppression of Xenopus Rx-L function impaired the formation of the photoreceptor layer and reduced the expression of photoreceptor specific genes. Overexpression of Xenopus Rx-L induced ectopic expression of photoreceptor specific genes, but did only marginally promote the proliferation of retinal progenitor cells. Targeted overexpression of Rx-L in developing retinoblasts in vivo led to an increased fraction of photoreceptor cells at the expense of amacrine and bipolar cells and revealed that Rx-L acts as a transcription activator. A phylogenic analysis of all reported Rx-type genes revealed that they could be grouped into three categories, including an "invertebrate Rx" group, a "classical vertebrate Rx" group, and a "vertebrate Qrx/Rx-L" group. Taken together, Rx-L, unlike Rx1, is required for the determination of retinal cell types, especially photoreceptors, rather than for proliferation of retinal progenitors. PMID- 19135437 TI - Design flexibility in cis-regulatory control of gene expression: synthetic and comparative evidence. AB - In early Drosophila embryos, the transcription factor Dorsal regulates patterns of gene expression and cell fate specification along the dorsal-ventral axis. How gene expression is produced within the broad lateral domain of the presumptive neurogenic ectoderm is not understood. To investigate transcriptional control during neurogenic ectoderm specification, we examined divergence and function of an embryonic cis-regulatory element controlling the gene short gastrulation (sog). While transcription factor binding sites are not completely conserved, we demonstrate that these sequences are bona fide regulatory elements, despite variable regulatory architecture. Mutation of conserved sequences revealed that putative transcription factor binding sites for Dorsal and Zelda, a ubiquitous maternal transcription factor, are required for proper sog expression. When Zelda and Dorsal sites are paired in a synthetic regulatory element, broad lateral expression results. However, synthetic regulatory elements that contain Dorsal and an additional activator also drive expression throughout the neurogenic ectoderm. Our results suggest that interaction between Dorsal and Zelda drives expression within the presumptive neurogenic ectoderm, but they also demonstrate that regulatory architecture directing expression in this domain is flexible. We propose a model for neurogenic ectoderm specification in which gene regulation occurs at the intersection of temporal and spatial transcription factor inputs. PMID- 19135438 TI - EphA4 and EfnB2a maintain rhombomere coherence by independently regulating intercalation of progenitor cells in the zebrafish neural keel. AB - During vertebrate development, the hindbrain is transiently segmented into 7 distinct rhombomeres (r). Hindbrain segmentation takes place within the context of the complex morphogenesis required for neurulation, which in zebrafish involves a characteristic cross-midline division that distributes progenitor cells bilaterally in the forming neural tube. The Eph receptor tyrosine kinase EphA4 and the membrane-bound Ephrin (Efn) ligand EfnB2a, which are expressed in complementary segments in the early hindbrain, are required for rhombomere boundary formation. We showed previously that EphA4 promotes cell-cell affinity within r3 and r5, and proposed that preferential adhesion within rhombomeres contributes to boundary formation. Here we show that EfnB2a is similarly required in r4 for normal cell affinity and that EphA4 and EfnB2a regulate cell affinity independently within their respective rhombomeres. Live imaging of cell sorting in mosaic embryos shows that both proteins function during cross-midline cell divisions in the hindbrain neural keel. Consistent with this, mosaic EfnB2a over expression causes widespread cell sorting and disrupts hindbrain organization, but only if induced at or before neural keel stage. We propose a model in which Eph and Efn-dependent cell affinity within rhombomeres serve to maintain rhombomere organization during the potentially disruptive process of teleost neurulation. PMID- 19135439 TI - Mitogen-activated protein kinase 3/mitogen-activated protein kinase 1 activates apoptosis during testicular ischemia-reperfusion injury in a nuclear factor kappaB-independent manner. AB - Nuclear factor kappa-B (NF-kappaB), mitogen-activated protein kinase3/MAPK1 and MAPK8 are involved in testicular ischemia reperfusion injury (testicular-I/R). NF kappaB knock-out mice (KO) subjected to testicular-I/R have a reduced testicular damage, blunted MAPK8 activation and enhanced MAPK3/MAPK1 activity. To better understand the role of MAPK3/MAPK1 up-regulation during testicular-I/R, we investigated the effects of PD98059, an inhibitor of MAPK3/MAPK1, in KO mice during testicular-I/R. KO and wild-type (WT) animals underwent 1 h testicular ischemia followed by 24 h reperfusion or a sham testicular-I/R. Animals received either PD98059 (5 mg/kg/ip) or its vehicle. MAPK3/MAPK1, BAX, caspase-3 and -9 and TNF-alpha expression were assessed along with histological examination and an immunostaining for protein of apoptosis. Testicular-I/R caused a greater increase in MAPK3/MAPK1 in KO than in WT animals in both testes. KO mice had a lower expression of the apoptotic proteins and TNF-alpha as well as reduced histological damage compared to WT. Immunostaining confirmed the lower expression of BAX in the Leydig cells of KO mice. Administration of PD98059, abrogated MAPK3/MAPK1 expression and slightly reduced TNF-alpha but did not improve or reverse the histological damage in KO. PD98059 significantly reduced the histological damage in WT mice and markedly reduced the apoptotic proteins in KO and WT mice. These results suggest that testicular-I/R triggers also a pathway of organ damage involving MAPK3/MAPK1, TNF-alpha, BAX, caspase-3 and -9 that activates an apoptotic machinery in an NF-kappaB independent manner. These findings should contribute to better understand testicular torsion-induced damage. PMID- 19135440 TI - Comparative therapeutic effects of metformin and vitamin E in a model of non alcoholic steatohepatitis in the young rat. AB - Only in the last few years has non-alcoholic steatohepatitis been recognized as an important and relatively common liver disease. To date, the therapeutic options are limited, vitamin E and metformin have been proposed for the treatment of this condition, although their mechanisms are not completely clarified as yet. The aim of this study was to investigate the anti-inflammatory and anti-oxidative mechanisms of these drugs in an experimental model of non-alcoholic steatohepatitis in the young rat. Male rats, just after weaning, were divided into four groups: a control group that received a standard diet; a high fat diet group; two high fat diet fed groups treated with vitamin E or metformin, respectively. After 4 weeks, we evaluated in the liver the modification of lipid peroxidation, assessed by malondialdehyde, TNF-alpha levels, S-nitrosylated protein, inducible nitric oxide sinthase (iNOS), and peroxisome proliferators activated receptors (PPAR) expression and metalloproteinase activity. High fat diet increased malondialdehyde, nitrotyrosilated proteins, and TNF-alpha tissue content. Moreover, a decrease of PPAR-alpha and an increase of PPAR-gamma expression were observed. An increase of metalloproteinase activity was also shown. Among drug treatments, metformin reduced body weight gain and fat mass, metalloproteinase activity, and TNF-alpha tissue content, while it restored PPAR alpha expression and downregulated PPAR-gamma expression. Vitamin E reduced the oxidative damage, protein nitrotyrosilation, and tissue TNF-alpha levels. Moreover a decrease of PPAR-gamma expression was also shown. These findings confirm the efficacy of both drugs as therapeutic tools in preventing the early onset of liver damage and non-alcoholic fatty liver disease progression. PMID- 19135441 TI - Comparative effects of bevacizumab, ranibizumab and pegaptanib at intravitreal dose range on endothelial cells. AB - Anti-VEGF therapy proved to be useful against several ocular pathological situations, including choroidal neovascularization and proliferative retinopathies. Ranibizumab (Ran), Pegaptanib (Peg) and Bevacizumab (Bev) are the pharmacological agents more frequently used in clinical practice by intravitreal injection. However, their exact effects on the angiogenic process have not been accurately established in a comparative study. The aim of the present study was to elucidate the precise effects of Ran, Peg and Bev on the multiple steps of the angiogenic process. Human umbilical vein endothelial cells (HUVEC) were incubated with each agent within the clinically established concentration range, or identical amounts of the excipients; cell cytotoxicity, proliferation, apoptosis, migration and vessel assembly were assessed. No cytotoxic effects were found for any of the agents studied at any concentration tested. At the clinical dose, cell proliferation was significantly reduced by Bev and Ran, whereas no difference was observed after Peg treatment. In addition, HUVEC apoptosis was effectively increased by Bev and Ran. Cell migration was reduced after incubation with every agent analyzed, though only reaching statistical significance upon Ran intravitreal dose. At clinical doses, capillary assembly was only affected by Bev. In agreement with these data, the active form of VEGF receptor-2 expression was decreased after incubation with Bev (to 66% of control values), Ran (78%) and Peg (86%) relative to controls. These findings indicate that these three agents display distinct effects on endothelial cells. PMID- 19135443 TI - Control of aquaporin 2 expression in collecting ducts of quail kidneys. AB - Birds and mammals are the only vertebrates that can concentrate urine. Avian kidneys contain structurally primitive loopless nephrons and also more advanced looped nephrons, in the cortical and medullary regions, respectively. We have identified the gene sequence of an aquaporin 2 (AQP2)-homologue water channel in collecting ducts of kidneys from adult quail, Coturnix japonica. Although immunoreactive quail AQP2 (qAQP2) was found in both types of nephrons, the expression is enhanced more clearly in the medullary regions after water deprivation. We therefore hypothesized that regulation of qAQP2 expression in quail kidneys via antidiuretic hormone (ADH) may require more advanced nephron structure. In this study, we determined the expression of qAQP2 mRNA in tissues isolated from the cortical and medullary regions before and after water deprivation, by conventional reverse transcriptase-polymerase chain reaction (RT PCR) and quantitative real-time PCR. In both normally hydrated and water-deprived groups, qAQP2 mRNA levels in the medullary regions were significantly higher (P<0.01) than in the cortical regions. In medullary areas, qAQP2 mRNA levels (real-time PCR normalized with 18S) were significantly higher (P<0.01, ANOVA) after water deprivation (1.09+/-0.10) than in normally hydrated controls (0.46+/ 0.08). In cortical areas, qAQP2 mRNA levels were also higher after water deprivation (0.37+/-0.05) than in controls (0.11+/-0.02). qAQP2 mRNA signals determined by in situ hybridization of digoxigenin-labeled riboprobe were also enhanced after water deprivation in both cortical and medullary collecting ducts. The results suggest that, contrary to our hypothesis, the endogenous production of ADH by water deprivation stimulates qAQP2 mRNA in both loopless and looped nephrons. PMID- 19135442 TI - Cholinergic degeneration and memory loss delayed by vitamin E in a Down syndrome mouse model. AB - Down syndrome (DS) individuals develop several neuropathological hallmarks seen in Alzheimer's disease, including cognitive decline and the early loss of cholinergic markers in the basal forebrain. These deficits are replicated in the Ts65Dn mouse, which contains a partial trisomy of murine chromosome 16, the orthologous genetic segment to human chromosome 21. Oxidative stress levels are elevated early in DS, and may contribute to the neurodegeneration seen in these individuals. We evaluated oxidative stress in Ts65Dn mice, and assessed the efficacy of long-term antioxidant supplementation on memory and basal forebrain pathology. We report that oxidative stress was elevated in the adult Ts65Dn brain, and that supplementation with the antioxidant vitamin E effectively reduced these markers. Also, Ts65Dn mice receiving vitamin E exhibited improved performance on a spatial working memory task and showed an attenuation of cholinergic neuron pathology in the basal forebrain. This study provides evidence that vitamin E delays onset of cognitive and morphological abnormalities in a mouse model of DS, and may represent a safe and effective treatment early in the progression of DS neuropathology. PMID- 19135444 TI - Identification of putative peptide paracrines/hormones in the water flea Daphnia pulex (Crustacea; Branchiopoda; Cladocera) using transcriptomics and immunohistochemistry. AB - The cladoceran crustacean Daphnia pulex has emerged as a model species for many biological fields, in particular environmental toxicology and toxicogenomics. Recently, this species has been the subject of an extensive transcriptome project, resulting in the generation and public deposition of over 150,000 expressed sequence tags (ESTs). This resource makes D. pulex an excellent model for protein discovery using bioinformatics. Here, in silico searches of the D. pulex EST database were conducted to identify transcripts encoding putative peptide precursors. Moreover, the mature peptides contained within the deduced prepro-hormones were predicted using online peptide processing programs and homology to known arthropod isoforms. In total, 63 putative peptide-encoding ESTs were identified encompassing 14 distinct peptide families/subfamilies: A-type allatostatin, B-type allatostatin, C-type allatostatin, bursicon (both alpha and beta subunit peptides), crustacean cardioactive peptide (CCAP), crustacean hyperglycemic hormone (CHH)/ion transport peptide (both CHH- and moult-inhibiting hormone-like subfamilies), diuretic hormone (calcitonin-like), ecdysis-triggering hormone (ETH), FMRFamide (both neuropeptide F and short neuropeptide F subfamilies), orcokinin and pigment dispersing hormone. From these transcripts, the structures of 76 full-length/partial peptides were predicted, which included the first C-type allatostatin-like peptide identified from a crustacean, the first crustacean calcitonin-like diuretic hormone, an undescribed CCAP isoform, two hitherto unknown ETH variants, and two new orcokinins. Neuronal localization of several of the identified peptide families was confirmed using immunohistochemitry (i.e. A-type allatostatin, CCAP, FMRFamide and PDH). In addition, immunohistochemical analyses identified other putative neuropeptides for which no ESTs had been found (i.e. corazonin, insect kinin, proctolin, red pigment concentrating hormone, SIFamide, sulfakinin and tachykinin-related peptide). Collectively, the data presented here not only catalog an extensive array of putative D. pulex peptide paracrines/hormones, but also provide a strong foundation for future investigations of the effects of environmental/anthropogenic stressors on peptidergic control in this model organism. PMID- 19135445 TI - Thyrotropin in teleost fish. AB - Thyrotropin (TSH), a pituitary glycoprotein hormone that stimulates the thyroid gland, has been cloned and sequenced from over a dozen teleost fish species. Although TSH is established as a primary driver of systemic thyroid status in mammals, its importance in the regulation of fish thyroid function is still uncertain. We review recent studies indicating that TSH structure is highly conserved across species representing six teleost families. These studies have found TSH messenger RNA consistently expressed in teleost pituitary tissue, although ectopic expression, particularly in gonads, has also been observed. They have also provided evidence for negative feedback inhibition of TSH expression by thyroid hormones, as well as stimulation by hypothalamic peptides. Descriptive studies have found increased TSHbeta expression associated with life history events thought to be promoted by thyroid hormones. These results, coupled with the discovery of a G-protein coupled TSH receptor in several teleost species, supports an active and conserved role for TSH in the regulation of teleost thyroid function. The relative importance of central pathways in regulating thyroid hormone provision to targets and the identity of a proposed thyrotropin inhibiting factor in teleost fish are still unanswered questions whose resolution will be facilitated by development of methods to measure circulating TSH and its secretion from the pituitary gland. PMID- 19135447 TI - Effect of peptide pools on effector functions of antigen-specific CD8+ T cells. AB - Peptide pools are routinely used to study antigen specific T cell responses, both in epitope discovery as well as immune monitoring. However, optimal assay conditions such as concentration of peptides or the best possible number of peptides per pool have not been defined. Thus, we examined whether different peptide concentrations or varying number of peptides per pool influence effector functions of antigen-specific human T-cells. PBMC isolated from HLA-A2-positive individuals with known responses to frequently recognised dominant CD8+ T cell epitopes derived from four different viruses (influenza virus, CMV, EBV, or HCV) were studied. PBMC were cultured with one of these HLA-A2 restricted peptides and varying concentrations of overlapping peptide pools derived from unrelated viruses specific for the hepatitis D and E viruses, the subjects have not been exposed to. Importantly, unrelated peptide pools inhibited the proliferation of IV-M1(58), CMVpp65(495-503), EBV-BMLF(1259-267) and HCV NS3(1073)-specific CD8 T cells in a dose dependent manner. Similarly, an increase in the number of peptides per pool also impaired antigen specific CD8+ T cell proliferation. In contrast, secretion of cytokines such as IL-2, IL-10, IFN-gamma, TNF-alpha or IP 10 as well as cytotoxicity was not affected by these unrelated peptide pools. The inhibition of proliferation could be restored by blocking PD-1/PDL-1 interaction and was not dependent on DMSO when DMSO concentration was 150). PMID- 19135477 TI - Functional selectivity of EGF family peptide growth factors: implications for cancer. AB - Breast, prostate, pancreatic, colorectal, lung, and head and neck cancers exploit deregulated signaling by ErbB family receptors and their ligands, EGF family peptide growth factors. EGF family members that bind the same receptor are able to stimulate divergent biological responses both in cell culture and in vivo. This is analogous to the functional selectivity exhibited by ligands for G protein coupled receptors. Here we review this literature and propose that this functional selectivity of EGF family members is due to distinctions in the conformation of the liganded receptor and subsequent differences in the sites of receptor tyrosine phosphorylation and receptor coupling to signaling effectors. We also discuss the roles of divergent ligand activity in establishing and maintaining malignant phenotypes. Finally, we discuss the potential of mutant EGF family ligands as cancer chemotherapeutics targeted to ErbB receptors. PMID- 19135479 TI - Enhancement of dendritic cell-based vaccine potency by anti-apoptotic siRNAs targeting key pro-apoptotic proteins in cytotoxic CD8(+) T cell-mediated cell death. AB - Dendritic cells (DCs) have become an important measure for the treatment of malignancies. Current DC preparations, however, generate short-lived DCs because they are subject to cell death from various apoptotic pressures. Antigen-specific CD8(+) cytotoxic T lymphocytes (CTLs) is one of the main obstacles to limit the DC-mediated immune priming since CTLs can recognize the target antigen expressing DCs as target cells and kill the DCs. CTLs secret perforin and serine protease granzymes during CTL killing. Perforin and serine protease granzymes induce the release of a number of mitochondrial pro-apoptotic factors, which are controlled by members of the BCL-2 family, such as BAK, BAX and BIM. FasL linking to Fas on DCs triggers the activation of caspase-8, which eventually leads to mitochondria mediated apoptosis via truncation of BID. In this study, we tried to enhance the DC priming capacity by prolonging DC survival using anti-apoptotic siRNA targeting these key pro-apoptotic molecules in CTL killing. Human papillomavirus (HPV)-16 E7 antigen presenting DCs that were transfected with these anti apoptotic siRNAs showed increased resistance to T cell-mediated death, leading to enhanced E7-specific CD8(+) T cell activation in vitro and in vivo. Among them, siRNA targeting BIM (siBIM) generated strongest E7-specific E7-specific CD8(+) T cell immunity. More importantly, vaccination with E7 presenting DCs transfected with siBIM was capable of generating a marked therapeutic effect in vaccinated mice. Our data indicate that ex vivo manipulation of DCs with siBIM may represent a plausible strategy for enhancing dendritic cell-based vaccine potency. PMID- 19135480 TI - A new generic real-time reverse transcription polymerase chain reaction assay for vesiviruses; vesiviruses were not detected in human samples. AB - Different viruses belonging to the genus Vesivirus infect a broad range of animals, and cause gastroenteritis, vesicular skin lesions, hemorrhagic disease, respiratory diseases and other conditions. A recent report on Vesivirus viremia, as detected by PCR, in samples from patients with hepatitis of unknown etiology in the USA suggested a zoonotic potential for vesiviruses. These results have not been confirmed by another laboratory. In order to do so, a generic PCR assay on the RNA polymerase region was developed, and validated with RNA from 69 different Vesivirus species. Except SMSV serotype-8, all species tested were detected, including the ones that were suggested to be involved in zoonotic transmission in the USA (SMSV serotype-5). The generic Vesivirus assay was used on RNA extracted from serum samples from patients with hepatitis, stool samples from patients with gastroenteritis, throat-swab specimens of patients with rash illnesses, throat swab and nose-swabs of patients with acute respiratory diseases, and cell cultures with cytopathologic effect from enterovirus surveillance in which no pathogen was found. None were found positive. In this study a generic Vesivirus assay was developed and it was concluded that vesiviruses are an unlikely cause of common illnesses in humans in the Netherlands. PMID- 19135481 TI - A new sensitive and automated chemiluminescent microparticle immunoassay for quantitative determination of hepatitis C virus core antigen. AB - A new sensitive and automated chemiluminescent assay was developed for the quantitative determination of hepatitis C virus (HCV) core antigen (Ag) in human sera or plasma: the Abbott ARCHITECT HCV Ag test. The assay sensitivity was determined by testing 10 commercial HCV seroconversion panels. Without exception, a positive result for HCV core Ag was observed before anti-HCV detection, resulting in an average reduction in the period between exposure and detection of 35.8 days. Both HCV core Ag and HCV RNA were detected in the panels at the same time, indicating equivalent sensitivity and detectability. A total of 197 HCV specimens comprising genotypes 1a, 1b, 2a, 2b, 3a, 3k, 4a, 5a and 6a were evaluated. Among these, 196 (99.5%), 191 (97%) and 193 (98%) were reactive using the HCV Ag, the immunoradiometric HCV Ag and the Amplicor HCV Monitor 2 assays, respectively. A comparison with the Amplicor HCV Monitor 2 showed a correlation coefficient (r) of 0.74. The specificity of the assay was established at 99.8% by testing 5403 specimens from US volunteer blood donors, hospitalized patients and individuals with medical conditions unrelated to HCV infection, in addition to specimens containing potentially interfering substances. PMID- 19135482 TI - Design, synthesis, molecular modeling, and anti-HIV-1 integrase activity of a series of photoactivatable diketo acid-containing inhibitors as affinity probes. AB - The diketo acid (DKA) class of HIV-1 integrase (IN) inhibitors is thought to function by chelating divalent metal ions on the enzyme catalytic site. However, differences in mutations conferring resistance to various DKA inhibitors suggest that multiple binding orientations may exist. In order to facilitate identification of DKA binding sites, a series of photoactivable analogues of two potent DKAs was prepared as novel photoaffinity probes. In cross-linking assays designed to measure disruption of substrate DNA binding, the photoprobes behaved similarly to a reference DKA inhibitor. Molecular modeling studies suggest that such photoprobes interact within the IN active site in a manner similar to that of the parent DKAs. Analogues Ia-c are novel photoaffinity ligands useful in clarifying the HIV-1 binding interactions of DKA inhibitors. PMID- 19135483 TI - Enhanced protective immunity against H5N1 influenza virus challenge by vaccination with DNA expressing a chimeric hemagglutinin in combination with an MHC class I-restricted epitope of nucleoprotein in mice. AB - DNA vaccination is an effective means of eliciting both humoral and cellular immune responses. The hemagglutinin (HA) surface protein of influenza A virus is a major target of protective antibody responses induced by virus infection or by vaccination and is widely considered to be the antigen of choice for an influenza vaccine. Cytotoxic T lymphocyte (CTL) responses directed against the conserved nucleoprotein (NP) are thought to play an important role in clearing virus and promoting survival and recovery from influenza. In this study, we developed a novel DNA vaccine approach using a chimeric plasmid consisting of the HA of H5N1 influenza virus in which an MHC class I-restricted NP-specific CTL epitope (NP147 155) was inserted. Immunogenicity and antiviral efficacy of this vaccine was assessed in mouse models. A similar level of HA expression was achieved in 293T cells transfected with pHA/NP(147-155) compared to that with pHA. Besides eliciting the specific anti-HA antibody responses, vaccination using pHA/NP(147 155) in mice induced NP epitope-specific CD8(+) T cell responses, which are generally not inducible by vaccination with pHA alone. After H5N1 influenza virus challenge, BALB/c mice vaccinated with pHA/NP(147-155) exhibited reduced inflammation severity and lung viral titers compared to those vaccinated with pHA. Our work may contribute to improvement of HA-based influenza DNA vaccines. PMID- 19135484 TI - Studying the effect of single mismatches in primer and probe binding regions on amplification curves and quantification in real-time PCR. AB - Comparison of a broad range of characteristics of real-time PCR amplification curves yielded only slight alterations for low numbers of mismatches in the primer binding regions, resulting in a quantification error up to 63.12%. The effects were more pronounced for mismatches in the probe binding region and resulted in a quantification error up to 33%. PMID- 19135485 TI - Alternative procedures for analysis of lipid A modification with phosphoethanolamine or aminoarabinose. AB - MALDI-TOF mass spectrometry analysis of lipid A prepared using a Tri-reagent based procedure with a 5-chloro-2-mercaptobenzothiazole matrix was preferable for the detection of phosphoethanolamine modification. In contrast, the analysis of lipid A prepared using an LPS extraction kit-based procedure with 2,5 dihydroxybenzoic acid was preferable for the detection of aminoarabinose modification. PMID- 19135486 TI - A spectralanalytic approach to emotional responses evoked through picture presentation. AB - Frontal EEG asymmetry has been linked to emotional and motivational reactivity. A frequently applied method to provoke specific asymmetry profiles is the presentation of affective film clips. Although these films might elicit strong emotional reactions, the exact time course and peak of an affective response remains unclear. In an alternative attempt, stimuli from the International Affective Picture System (IAPS), known to reliably alter emotional states, are utilized. These stimuli are less likely to cause excessive variations in affective responding. However, relevant studies have most often been unable to find the predicted effects. One reason for such failures might be the inadequate knowledge about the minimum number of stimuli needed for psychometrically stable results. In the present study, an adequate split-half reliability for the experimental procedure was assured and substantial effects of affective picture category were found. This pattern of results was robust for both Cz and linked ears as reference. Thus, presenting pictures with an adequate recording length might be a reliable alternative for inducing affective reactions in alpha asymmetry research. PMID- 19135487 TI - Binding of proteins to PEGylated liposomes and improvement of G-CSF efficacy in mobilization of hematopoietic stem cells. AB - We have previously shown that formulation of coagulation factor VIII and activated factor VII with PEGylated liposomes (PEGLip) results in an extension of circulation time and an increase in hemostatic efficacy. Here we identified additional proteins that associate with PEGLip, including granulocyte colony stimulating factor (G-CSF). Surface plasmon resonance analyses indicated that G CSF bound noncovalently but with high affinity and specificity to PEGLip. A pharmacokinetic study in mice demonstrated that PEGLip formulation of G-CSF extended its circulation time and resulted in higher G-CSF levels several hours after both subcutaneous and intravenous injection. PEGLip-formulated G-CSF had a significantly improved efficacy in the mobilization of hematopoietic stem cells (HSC) from the bone marrow to the peripheral blood. The results suggest that PEGLip-formulated G-CSF may function as an effective and safe tool for the mobilization of HSC prior to bone marrow transplantation. We also identified an amino acid sequence present in proteins that associate with PEGLip but absent from those that do not. A peptide based on this consensus sequence bound PEGLip. The results suggest that PEGLip formulation may serve as a platform for the delivery of additional short-half-life proteins/peptides having the relevant consensus sequence. PMID- 19135489 TI - Longitudinal relationships between fathers', mothers', and adolescents' restrained eating. AB - Family members can exert important direct and indirect influence on the eating behaviours of children; these include modelling, and transmission of preferences, beliefs, and attitudes. Current studies on family similarities in dieting behaviours however show inconsistent results. The present study examines family similarities and reciprocal influences on restrained eating, using data of a longitudinal three-wave full-family study consisting of both parents and two adolescent sibling children (aged 13-16 at time 1) from 404 Dutch families. All family members reported their restrained eating behaviours at three annual waves. Cross-sectional associations were found between parents' and adolescents' restrained eating, but overall no transmission of restrained eating was found between family members over time. Similarities were higher between daughters and mothers compared to sons and mothers. Longitudinally, no differences in the results were found between boys and girls, or between adolescents with high or low quality relationships with their parents. PMID- 19135488 TI - Mathematical modeling of molecular diffusion through mucus. AB - The rate of molecular transport through the mucus gel can be an important determinant of efficacy for therapeutic agents delivered by oral, intranasal, intravaginal/rectal, and intraocular routes. Transport through mucus can be described by mathematical models based on principles of physical chemistry and known characteristics of the mucus gel, its constituents, and of the drug itself. In this paper, we review mathematical models of molecular diffusion in mucus, as well as the techniques commonly used to measure diffusion of solutes in the mucus gel, mucus gel mimics, and mucosal epithelia. PMID- 19135490 TI - Sympathetic nervous system-targeted neuropeptide Y overexpression in mice enhances neointimal formation in response to vascular injury. AB - Sympathetic neurotransmitter neuropeptide Y (NPY) is associated with vascular remodelling, neointimal hyperplasia and atherosclerosis in experimental animal models and clinical studies. In order to study the role of sympathetic nerve produced NPY in vascular diseases, transgenic mouse model overexpressing NPY in central and peripheral noradrenergic neurons under the dopamine-beta-hydroxylase (DBH) promoter was recently created (OE-NPY(DBH) mouse). This study aimed to examine the effect of NPY overexpression on arterial neointimal hyperplasia in an experimental model of vascular injury. Transgenic OE-NPY(DBH) mice and wildtype control mice of two different inbred strains (C57BL/6 and FVB/n) underwent a femoral artery surgery with a transluminar injury by a 0.38-mm guide wire insertion. Arteries were harvested 4 weeks from the surgery, and they were stained for basic morphology. Both strains of OE-NPY(DBH) mice, as compared with wildtype control mice, showed on average 50% greater formation of the neointima (P<0.01) and an increase in the medial area (P=0.05). The results suggest that moderately increased neuronal NPY causes the arteries to be more susceptible to femoral artery thickening after endothelial injury. The OE-NPY(DBH) mouse provides a novel tool to explore the role of NPY in the development of vascular disease related to metabolic disorders. PMID- 19135491 TI - Cloning, tissue distribution and effects of food deprivation on pituitary adenylate cyclase activating polypeptide (PACAP)/PACAP-related peptide (PRP) and preprosomatostatin 1 (PPSS 1) in Atlantic cod (Gadus morhua). AB - Full-length complementary deoxyribonucleic acid sequences encoding pituitary adenylate cyclase activating polypeptide (PACAP)/PACAP-related peptide (PRP) and preprosomatostatin 1 (PPSS 1) were cloned from Atlantic cod (Gadus morhua) hypothalamus using reverse transcription and rapid amplification of complementary deoxyribonucleic acid ends. Semi-quantitative reverse transcriptase polymerase chain reaction shows that PRP/PACAP mRNA and PPSS 1 mRNA are widely distributed throughout cod brain. During development, PRP/PACAP and PPSS 1 were detected at the 30-somite stage and pre-hatching stage, respectively, and expression levels gradually increased up to the hatched larvae. PPSS 1, but not PRP/PACAP, appeared to be affected by food availability during early development. In juvenile cod, PPSS 1 expression levels increased and remained significantly higher than that of control fed fish throughout 30 days of starvation and during a subsequent 10 days refeeding period. In contrast, PRP/PACAP expression levels were not affected by 30 days of food deprivation, but a significant increase in expression levels was observed during the 10 days refeeding period in the experimental food-deprived group as compared to the control fed group. Our results suggest that PRP/PACAP and PPSS 1 may be involved in the complex regulation of growth, feeding and metabolism during food deprivation and refeeding in Atlantic cod. PMID- 19135492 TI - Estimating the cost-effectiveness of vaccination against herpes zoster in England and Wales. AB - A live-attenuated vaccine against herpes zoster (HZ) has been approved for use, on the basis of a large-scale clinical trial that suggests that the vaccine is safe and efficacious. This study uses a Markov cohort model to estimate whether routine vaccination of the elderly (60+) would be cost-effective, when compared with other uses of health care resources. Vaccine efficacy parameters are estimated by fitting a model to clinical trial data. Estimates of QALY losses due to acute HZ and post-herpetic neuralgia were derived by fitting models to data on the duration of pain by severity and the QoL detriment associated with different severity categories, as reported in a number of different studies. Other parameters (such as cost and incidence estimates) were based on the literature, or UK data sources. The results suggest that vaccination of 65 year olds is likely to be cost-effective (base-case ICER=pound20,400 per QALY gained). If the vaccine does offer additional protection against either the severity of disease or the likelihood of developing PHN (as suggested by the clinical trial), then vaccination of all elderly age groups is highly likely to be deemed cost effective. Vaccination at either 65 or 70 years (depending on assumptions of the vaccine action) is most cost-effective. Including a booster dose at a later age is unlikely to be cost-effective. PMID- 19135493 TI - Knowledge and attitudes about human papillomavirus (HPV) and HPV vaccines among women living in metropolitan and rural regions of China. AB - Infection with the human papillomavirus (HPV) is one of the most common sexually transmitted infections and causes virtually all cervical cancer globally. The recent development of two safe and clinically effective vaccines against HPV is a promising step towards lowering cervical cancer rates in the future. What Chinese women think about HPV and the vaccines remains unknown. We undertook a population based survey, which was embedded in a cervical cancer screening project and was designed to assess women's knowledge about HPV and their acceptability to the vaccines. We found that only 15.0% of women in our study reported to have ever heard of HPV, and this knowledge differs by rural (9.3%) and metropolitan areas (21.6%) and also by education. Most (84.6%) participants were willing to be vaccinated if HPV vaccine became available to them. The present study documents ways in which women learn about HPV and indicates the potential barriers and success of introducing HPV vaccine to China. PMID- 19135494 TI - Factors associated with incomplete vaccination of babies at risk of perinatal hepatitis B transmission: a London study in 2006. AB - We measured the hepatitis B (HB) vaccination uptake in 249 London babies born in 2004 to HBsAg positive mothers. Two thirds (69%) received three vaccinations and half (49%, 95% CI 43-56) received a complete course (four doses). Complete immunization was associated with sector of delivery (p<0.001), recording of the GP details in case notes, having booked for antenatal care, having a good command of English, and receipt of written information on HB. A third of the babies (33%) had a post-vaccination test; when the mother had other children, 39% of the oldest children were vaccinated; information on partner's vaccination was available for 12%. This study highlights that appropriate counseling and information should be provided to the mothers, and the importance in London of arrangements for integrated care across acute and primary care services. PMID- 19135495 TI - Protection against Yersinia pseudotuberculosis infection conferred by a Lactococcus lactis mucosal delivery vector secreting LcrV. AB - Herein, we sought to evaluate the potential of a recombinant Lactococcus lactis strain secreting the Yersinia pseudotuberculosis low-calcium response V (LcrV) antigen for mucosal vaccination against Yersinia infections. We showed that the recombinant strain induced specific systemic and mucosal antibody and cellular immune responses after intranasal immunization and protected mice against both oral and systemic Y. pseudotuberculosis infections. This constitutes the first proof of principle for a novel anti-Yersinia mucosal vaccination strategy using recombinant lactic acid bacteria. PMID- 19135496 TI - Safety and immunogenicity of SC599, an oral live attenuated Shigella dysenteriae type-1 vaccine in healthy volunteers: results of a Phase 2, randomized, double blind placebo-controlled trial. AB - SC599 vaccine is a live Shigella dysenteriae 1 strain attenuated by deletion of invasion [icsA], iron chelation [ent, fep] and shiga toxin A subunit [stxA] genes. In a preliminary Phase 1 single dose prospective study, we showed that SC599 vaccine was well tolerated, and the maximum tolerable dose was greater than 10(8) CFU [Sadorge C, Ndiaye A, Beveridge N, Frazer S, Giemza R, Jolly N, et al. Phase 1 clinical trial of live attenuated Shigella dysenteriae type-1 DeltaicsA Deltaent Deltafep DeltastxA:HgR oral vaccine SC599 in healthy human adult volunteers. Vaccine 2008; 26(7):978-8]. In this Phase 2 trial, three groups of volunteers ingested a single dose of SC599 [10(5) CFU, n=38; 10(7) CFU, n=36] or placebo [n=37]. Both 10(5) and 10(7) CFU doses were immunogenic, inducing significant IgA and IgG LPS-specific ASCs and antibody responses, comparable in magnitude to those of other strains that prevented illness following experimental challenge. In the intention to treat analysis, 34.2% and 44.4% IgA ASC responders were detected in the 10(5) and 10(7) CFU groups respectively (p<0001 vs placebo for both groups), as well as 31.6% and 33.3% serum IgA responders (p<001 and p<0.001 vs placebo for 10(5) and 10(7) CFU groups, respectively). No difference between the two vaccine groups was observed. No stxB-specific antibody response was detected in the vaccines. SC599 excretion occurred in 23.7 and 30.6% of subjects in the 10(5) and 10(7) CFU groups, respectively. SC599 vaccine was well tolerated, and the reported adverse events were mainly digestive. These results indicate that a single oral immunization of SC599 vaccine elicits a significant circulating IgA ASC and serum antibody response that may confer protection against the most severe symptoms of Shigellosis in responders to the vaccine. PMID- 19135498 TI - Chemical and toxicological characterization of commercial smokeless tobacco products available on the Canadian market. AB - Some health experts are recommending that smokers who refuse to quit or refuse to use nicotine replacement therapy (NRT) such as nicotine-containing chewing gum switch to certain types of smokeless tobacco products (STP) such as Swedish snus. Other health experts disagree citing the uncertainty in the composition of commercially available STP, the lack of governmental regulations to ensure that STP advertised to meet certain standards (i.e., GothiaTek) do actually meet such standards, and the uncertainty that any STP can provide as safe as alternative to smoking as NRT. One reason for uncertainty is the dearth of detailed chemical and toxicological information on contemporary STP. Unlike the situation with cigarettes, there are few standardized methods for analytical and toxicological studies of STP. Consequently, the objective for this work was to characterize several types of STP available on the Canadian market using the modifications of the Official Health Canada chemical and toxicological methods developed for cigarettes. Moist snuff samples tested had TSNA and B[a]P levels somewhat above the GothiaTek standard while samples of Swedish snus, low-moisture snuff, and US style chewing tobacco did not. Use of in vitro assays to assess STP toxicity was of limited utility in distinguishing product types. PMID- 19135497 TI - Efficacy and immunogenicity of Mycobacterium bovis DeltaRD1 against aerosol M. bovis infection in neonatal calves. AB - An attenuated Mycobacterium bovisRD1 deletion (DeltaRD1) mutant of the Ravenel strain was constructed, characterized, and sequenced. This M. bovis DeltaRD1 vaccine strain administered to calves at 2 weeks of age provided similar efficacy as M. bovis bacillus Calmette Guerin (BCG) against low dose, aerosol challenge with virulent M. bovis at 3.5 months of age. Approximately 4.5 months after challenge, both DeltaRD1- and BCG-vaccinates had reduced tuberculosis (TB) associated pathology in lungs and lung-associated lymph nodes and M. bovis colonization of tracheobronchial lymph nodes as compared to non-vaccinates. Mean central memory responses elicited by either DeltaRD1 or BCG prior to challenge correlated with reduced pathology and bacterial colonization. Neither DeltaRD1 or BCG elicited IFN-gamma responses to rESAT-6:CFP-10 prior to challenge, an emerging tool for modern TB surveillance programs. The DeltaRD1 strain may prove useful for bovine TB vaccine programs, particularly if additional mutations are included to improve safety and immunogenicity. PMID- 19135499 TI - Nicotine contents in various toothpowders (dant manjans): measurement and safety evaluation. AB - The use of tobacco products as dentrifice is prevalent in various parts of India. Among them toothpowder (dant manjan) is very common. These nicotine containing toothpastes/toothpowders are health hazards and is also habit forming. Health experts of India rightly banned use of nicotine containing toothpowder as early as 1992 by making proper legislation. We just made an attempt to verify whether the manufacturers complying the legislation or not. Eight leading brands of toothpowders were analyzed qualitatively by gas chromatography-mass spectrum detector and also quantitatively by gas chromatography-nitrogen phosphorus detector. Our results indicated four brands were found to contain nicotine in the range of 2.53 microg/g to 11.50 mg/g of toothpowder. This finding further confirms that addition of nicotine in dentifrice violates the regulatory norms. Regulatory authorities should give more attention to ensure that all toothpowders are free from nicotine which is also a statutory requirement. PMID- 19135501 TI - The use of OpdA in rat models of organic phosphorus (OP) poisoning. PMID- 19135502 TI - Structural and functional properties of Bp-LAAO, a new L-amino acid oxidase isolated from Bothrops pauloensis snake venom. AB - An L-amino acid oxidase (Bp-LAAO) from Bothrops pauloensis snake venom was highly purified using sequential chromatography steps on CM-Sepharose, Phenyl-Sepharose CL-4B, Benzamidine Sepharose and C18 reverse-phase HPLC. Purified Bp-LAAO showed to be a homodimeric acidic glycoprotein with molecular weight around 65kDa under reducing conditions in SDS-PAGE. The best substrates for Bp-LAAO were L-Met, L Leu, L-Phe and L-Ile and the enzyme showed a strong reduction of its catalytic activity upon L-Met and L-Phe substrates at extreme temperatures. Bp-LAAO showed leishmanicidal, antitumoral and bactericidal activities dose dependently. Bp-LAAO induced platelet aggregation in platelet-rich plasma and this activity was inhibited by catalase. Bp-LAAO-cDNA of 1548bp codified a mature protein with 516 amino acid residues corresponding to a theoretical isoelectric point and molecular weight of 6.3 and 58kDa, respectively. Additionally, structural and phylogenetic studies identified residues under positive selection and their probable location in Bp-LAAO and other snake venom LAAOs (svLAAOs). Structural and functional investigations of these enzymes can contribute to the advancement of toxinology and to the elaboration of novel therapeutic agents. PMID- 19135503 TI - Absence of age-related changes in nigral dopaminergic neurons of Asian Indians: relevance to lower incidence of Parkinson's disease. AB - Age-related loss of melanized nigral neurons reported in the British Caucasians is not observed in Asian Indian, American and French adults. In the Americans, loss of dopaminergic phenotype occurs from midlife, without frank neurodegeneration. Here, we investigated whether nigral dopaminergic neurons in Asian Indians are lost with age or undergo morphological or biochemical dysfunction. Using unbiased stereology we estimated volume, number of melanized, borderline/non-melanized (n=34, 28 gestational weeks to 80 years) and tyrosine hydroxylase (TH)-Nurr1 co-labeled neurons (n=32, 28 gestational weeks to 80 years) in substantia nigra pars compacta. We quantified Nurr1 and TH proteins by immunoblotting (n=18, 28 gestational weeks to 69 years) and apoptotic neurons by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining. Nuclear and soma size was estimated by morphometry. There was no age related decline in volume, neuronal density, neuronal numbers and TH-Nurr1 co labeled neurons. TH and Nurr1 protein expression remained stable. Lack of TUNEL TH co-labeled cells confirmed absence of neuronal apoptosis. The neuronal size remained unaltered. Our findings of preserved nigral dopaminergic neurons suggest no age-related loss of nigral function in Asian Indians, unlike the Americans. This may explain the lower incidence of Parkinson's disease in Asian Indians. PMID- 19135504 TI - Acid-sensitive TASK-like K+ conductances contribute to resting membrane potential and to orexin-induced membrane depolarization in rat thalamic paraventricular nucleus neurons. AB - Orexin (hypocretin) peptides are known to depolarize rat thalamic paraventricular nucleus (PVT) neurons by suppression of one or more undefined potassium conductances. Here, we investigated a contribution of TWIK-related acid-sensitive K(+) (TASK) channels to the resting membrane potential and orexin-induced depolarization of PVT neurons, using patch clamp recording techniques in brain slice preparations. Upon exposure to an acidic (pH 6.3) extracellular solution, PVT neurons displayed membrane depolarization. Under voltage-clamp and in the presence of tetrodotoxin (TTX, 0.5 microM), low pH solutions induced an inward shift in baseline membrane current, accompanied by a net decrease in membrane conductance, reversing close to the potassium equilibrium potential. By contrast, exposure to alkaline (pH 8.3) solutions resulted in membrane hyperpolarization, induced an outward shift in baseline membrane current and an increase in net conductance that reversed close to the potassium equilibrium potential. A local anesthetic bupivacaine (20-40 microM) and the endocannabinoid anandamide (5-10 microM) mimicked the effects of the acidic solution. Exposure to the volatile anesthetic isoflurane (0.2-0.5 mM) induced changes in resting membrane potential, baseline current and membrane conductance similar to those caused by the alkaline solution. Although responsiveness to orexins was preserved under each of the above conditions, the amplitude of the orexin B (0.5 microM)-induced inward current was depressed in the acidic solution and in the presence of anandamide, remained largely unchanged in the alkaline solution, and was enhanced by isoflurane when compared with that in normal artificial cerebrospinal solution. We conclude that pH-sensitive potassium channels, TASK-1 and TASK-3 channels, contribute substantially to the resting membrane conductance(s) and excitability in PVT neurons. The observations that orexin-induced currents were affected by putative TASK-specific drugs in a manner predictable from their effects on TASK channels also suggest that the orexin-induced excitation in PVT neurons is mediated by closure of TASK channels. PMID- 19135505 TI - Intracellular Zn2+ increases contribute to the progression of excitotoxic Ca2+ increases in apical dendrites of CA1 pyramidal neurons. AB - Sustained intracellular Ca(2+) elevation is a well-established contributor to neuronal injury following excessive activation of N-methyl-d-aspartic acid (NMDA) type glutamate receptors. Zn(2+) can also be involved in excitotoxic degeneration, but the relative contributions of these two cations to the initiation and progression of excitotoxic injury is not yet known. We previously concluded that extended NMDA exposure led to sustained Ca(2+) increases that originated in apical dendrites of CA1 neurons and then propagated slowly throughout neurons and caused rapid necrotic injury. However the fluorescent indicator used in those studies (Fura-6F) may also respond to Zn(2+), and in the present work we examine possible contributions of Zn(2+) to indicator signals and to the progression of degenerative signaling along murine CA1 dendrites. Selective chelation of Zn(2+) with N,N,N',N'-tetrakis(2 pyridylmethyl)ethylenediamine (TPEN) significantly delayed, but did not prevent the development and progression of sustained high-level Fura-6F signals from dendrites to somata. Rapid indicator loss during the Ca(2+) overload response, which corresponds to rapid neuronal injury, was also not prevented by TPEN. The relationship between cytosolic Zn(2+) and Ca(2+) levels was assessed in single CA1 neurons co-loaded with Fura-6F and the Zn(2+)-selective indicator FluoZin-3. NMDA exposure resulted in significant initial increases in FluoZin-3 increases that were prevented by TPEN, but not by extracellular Zn(2+) chelation with Ca EDTA. Consistent with this result, Ca-EDTA did not delay the progression of Fura 6F signals during NMDA. Removal of extracellular Ca(2+) reduced, but did not prevent FluoZin-3 increases. These results suggest that sustained Ca(2+) increases indeed underlie Fura-6F signals that slowly propagate throughout neurons, and that Ca(2+) (rather than Zn(2+)) increases are ultimately responsible for neuronal injury during NMDA. However, mobilization of Zn(2+) from endogenous sources leads to significant neuronal Zn(2+) increases, that in turn contribute to mechanisms of initiation and progression of progressive Ca(2+) deregulation. PMID- 19135506 TI - High dietary consumption of trans fatty acids decreases brain docosahexaenoic acid but does not alter amyloid-beta and tau pathologies in the 3xTg-AD model of Alzheimer's disease. AB - Dietary consumption of trans fatty acids (TFA) has increased during the 20th century and is a suspected risk factor for cardiovascular diseases. More recently, high TFA intake has been associated with a higher risk of developing Alzheimer's disease (AD). To investigate the impact of TFA on an animal model genetically programmed to express amyloid-beta (Abeta) and tau pathological markers of AD, we have fed 3xTg-AD mice with either control (0% TFA/total fatty acid), high TFA (16% TFA) or very high TFA (43% TFA) isocaloric diets from 2 to 16 months of age. Effects of TFA on plasma hepatic enzymes, glucose and lipid profile were minimal but very high TFA intake decreased visceral fat of non transgenic mice. Importantly, dietary TFA increased brain TFA concentrations in a dose-related manner. Very high TFA consumption substantially modified the brain fatty acid profile by increasing mono-unsaturated fatty acids and decreasing polyunsaturated fatty acids (PUFA). Very high TFA intake induced a shift from docosahexaenoic acid (DHA, 22:6n-3) toward n-6 docosapentaenoic acid (DPA, 22:5n 6) without altering the n-3:n-6 PUFA ratio in the cortex of both control and 3xTg AD mice. Changes in levels of Abeta(40), Abeta(42), tau protein, phosphorylated tau protein and synaptic markers were not statistically significant in the three groups of 3xTg-AD mice, despite a trend toward decreased insoluble tau in very high TFA-fed 3xTg-AD animals. In summary, TFA intake modulated brain fatty acid profiles but had no significant effect on major brain neuropathological hallmarks of AD in an animal model. PMID- 19135507 TI - Wnt signaling controls the fate of mesenchymal stem cells. AB - Multipotential mesenchymal stem cells (MSCs) are able to differentiate along several known lineages and have been shown to be efficacious for in vivo wound repair. The growth and differentiation of MSCs are known to be tightly regulated via interactions with specific extracellular mediators. Recent studies have shown that Wnts and their downstream signaling pathways play an important role in the self-renewal and differentiation of MSCs. Indeed altered bone-mass is known to result from mutations in LRP5, a Wnt co-receptor, that suggests Wnt plays an important signaling role during bone formation, possibly involving MSCs. This review outlines the current understanding of the distinct Wnt intracellular pathways including both canonical beta-catenin/TCF(LEF1) signaling and non canonical cascades mediated by JNK, PKC, Ca(2+) or Rho, and how they are involved in the regulation of MSC proliferation and differentiation. We also discuss the coordination between different Wnt signaling cascades to precisely control MSC cell fate decisions, and we dissect the functional cross-talk of Wnt signaling that is known to occur with other growth factor signaling pathways. PMID- 19135508 TI - Cold-adapted Antarctic fish: the discovery of neuroglobin in the dominant suborder Notothenioidei. AB - Novel globins, such as neuroglobin (Ngb) and cytoglobin, have recently been discovered in many vertebrates. Ngb is mainly expressed in neurons and plays a neuroprotective role during hypoxic stress. Neuronal hypoxia and cerebral ischemia induce Ngb expression; knocking down Ngb expression increases hypoxic neuronal injury in vitro and ischemic cerebral injury in vivo. Although Ngb was originally identified in mammals, it is also present in fish, including the zebrafish Danio rerio. We have discovered the Ngb gene to be present in red blooded notothenioid fish species, and in at least 13 of the 16 species of the white-blooded icefish family Channichthyidae. The deduced amino-acid sequences are well conserved. The retention of the Ngb gene by channichthyids, despite the loss of hemoglobin and myoglobin, appears very intriguing. PMID- 19135509 TI - Studies on the in vitro genotoxicity of baytubes, agglomerates of engineered multi-walled carbon-nanotubes (MWCNT). AB - The increasing production and expanding application of nanoparticles in multiple aspects of life necessitate reliable safety assessment. In this context we here report on the evaluation of the potential genotoxicity of baytubes, i.e. agglomerates of multi-walled carbon-nanotubes (MWCNT). Testing for chromosome aberrations was done in V79 cells and for gene mutations in the Salmonella microsome test. Baytubes were formulated in deionised water at 10 mg/ml and treated with ultrasound for 30 min at 25 degrees C. Particle size distribution was determined under the incubation conditions in the in vitro studies. In the chromosome aberration test V79 cells (OECD TG 473) were exposed in the absence or presence of S9 mix for 4 h to concentrations of 2.5, 5 and 10 microg/ml of baytubes (visible from concentration of 5 microg/ml and higher). Harvest was 18 h after the beginning of the treatment. In addition, cells treated with 10 microg/ml were harvested 30 h after the beginning of the treatment. An additional experiment was performed using continuous treatment at 2.5, 5 and 10 microg/ml for 18 h (no S9 mix) with subsequent harvest. Under these conditions and in the concentration range tested there were no cytotoxic and no clastogenic effects. In the Salmonella microsome (Ames) test (OECD TG 471) concentrations up to 5000 microg/plate were tested in Salmonella typhimurium (strains TA 1535, TA 100, TA 1537, TA 98 and TA 102) in the absence or presence of S9 mix. Under these conditions and in the concentration range tested there were no bacteriotoxic and no mutagenic effects. PMID- 19135510 TI - Suppressive effect of tobacco smoke extracts on oral P-glycoprotein function and its impact in smoke-induced insult to oral epidermal cells. AB - P-glycoprotein (Pgp) participates in the export of numerous toxins, drugs, and physiological compounds. To examine the involvement of Pgp in smoke-induced oral cell insult, the effects of extracts of the mainstream tobacco smoke (TS) on Pgp were studied in an oral epidermal carcinoma cell line, OECM-1. TS was first extracted with cyclohexane (CTS) and the residues were further extracted with isopropanol (ITS). For comparison, cells were exposed to CTS and ITS at the concentrations according to their relative extraction yield. ITS but not CTS decreased the efflux of a Pgp substrate, rhodamine (Rh) 123, in a concentration- and time-dependent manner. The efflux was also decreased by co-exposure to CTS and ITS. However, immunoblot analysis revealed that the protein level of Pgp was not affected by ITS. Naphthalene, mainly detected in the ITS, decreased Rh 123 efflux. However, the efflux activity was not affected by benzo(a)pyrene and nicotine, which were present in the CTS and both extracts, respectively. Co exposure to CTS in combination with ITS, naphthalene, or verapamil enhanced cell insult compared to single exposure. These results demonstrated that smoke and its constituent, naphthalene, diminished Pgp-mediated efflux. The reduction in Pgp function could be a stimulatory factor of TS-induced oral cell insult. PMID- 19135511 TI - Novel polymeric film coatings for colon targeting: how to adjust desired membrane properties. AB - The major aim of this work was to optimize the properties of novel polymeric films based on blends of ethylcellulose and Nutriose (a water-soluble, branched dextrin). Such blends were recently shown to be highly promising for the site specific delivery of drugs to the colon in patients suffering from inflammatory bowel diseases, in particular Crohn's disease and ulcerative colitis. Importantly, and in contrast to various other colon targeting approaches, the system is adapted to the pathophysiological conditions in the disease state. However, it is yet unknown how desired membrane properties, especially water uptake and dry mass loss kinetics as well as mechanical stability can be adjusted to the specific needs of particular drug treatments. Different highly efficient and easy to apply tools were identified altering the membrane's properties, in particular their mechanical resistance required to withstand the shear forces resulting from the motility of the upper GIT and the hydrostatic pressure built up within the devices upon contact with aqueous media. This includes the variation of the Nutriose:ethylcellulose blend ratio and initial plasticizer content. Importantly, Nutriose also exhibits significant pre-biotic activity, normalizing the microflora in the patients' colon, which is of major clinical benefit in the case of inflammatory bowel diseases. PMID- 19135512 TI - Injectable in situ forming depot systems: PEG-DAE as novel solvent for improved PLGA storage stability. AB - Injectable in situ forming depots (ISFD) that contain a peptide or a protein within a polymeric solution comprise an attractive, but challenging application system. Beyond chemical compatibility, local tolerability and acute toxicity, an important factor for an ISFD is its storage stability as a liquid. In this study, poly(D,L-lactide-co-glycolide) (PLGA) degradation in the presence of poly(ethyleneglycol) (PEG) as biocompatible solvent was investigated as a function of storage temperature and water content. The PLGA molecular weight (Mw) was determined by gel permeation chromatography (GPC), and monitored by NMR during degradation. Rapid PLGA degradation of 75% at 25 degrees C storage temperature was shown to be the result of a transesterification using conventional PEG as solvent. A significant improvement with only 3% Mw loss was obtained by capping the PEG hydroxy- with an alkyl- endgroup to have poly(ethyleneglycol) dialkylether (PEG-DAE). The formation of PEG-PLGA block co polymers was confirmed by NMR, only for PEG300. Reaction rate constants were used to compare PLGA degradation dissolved in conventional and alkylated PEGs. The degradation kinetics in PEG-DAE were almost completely insensitive to 1% additional water in the solution. The transesterification of the hydroxy endgroups of PEG with PLGA was the major degradation mechanism, even under hydrous conditions. The use of PEG-DAE for injectable polymeric solutions, showed PLGA stability under the chosen conditions for at least 2 months. Based on the results obtained here, PEG-DAE appears to be a promising excipient for PLGA based, parenteral ISFD. PMID- 19135513 TI - Preparation of oxidized sodium alginate-graft-poly((2-dimethylamino) ethyl methacrylate) gel beads and in vitro controlled release behavior of BSA. AB - Graft copolymerization of amino group-terminated poly((2-dimethylamino) ethyl methacrylate) (PDMAEMA-NH2) onto oxidized sodium alginate (OSA) was reacted without using a catalyst. The structure of the graft was investigated by Fourier transform infrared (FT-IR) spectroscopy. The OSA-g-PDMAEMA gel beads were prepared by dropping the aqueous solution of the graft copolymer into CaCl2 aqueous solution. The effects of pH and ionic strength on the swelling behaviors of the gel beads were studied. The results indicate that the gel beads have pH and ionic strength sensitivity. Bovine serum albumin (BSA) was entrapped in the beads and the in vitro drug release profiles were established in buffer solution with pH 1.8 (HCl), pH 7.4 (KH2PO4-NaOH), and 0.9% (w/v) NaCl at 37 degrees C. The results showed that the oral delivery of proteins can be controlled by adjusting the graft percentage (G, %), pH and ionic strength. According to this study, the OSA-g-PDMAEMA gel beads could be suitable for the oral delivery of proteins. PMID- 19135514 TI - Intravesical cationic nanoparticles of chitosan and polycaprolactone for the delivery of Mitomycin C to bladder tumors. AB - Cationic nanoparticles of chitosan (CS), poly-epsilon-caprolactone coated with chitosan (CS-PCL) and poly-epsilon-caprolactone coated with poly-L-lysine (PLL PCL) were developed to encapsulate intravesical chemotherapeutic agent Mitomycin C (MMC) for longer residence time, higher local drug concentration and prevention of drug loss during bladder discharge. Nanoparticle diameters varied between 180 and 340 nm depending on polymer used for preparation and coating. Zeta potential values demonstrated positive charge expected from cationic nanoparticles. MMC encapsulation efficiency depended on hydrophilicity of polymers since MMC is water-soluble. Encapsulation was increased by 2-fold for CS-PCL and 3-fold for PLL-PCL as a consequence of hydrophilic coating. Complete drug release was obtained with only CS-PCL nanoparticles. On the other hand, CS and PLL-PCL nanoparticles did not completely liberate MMC due to strong polymer-drug interactions which were elucidated with DSC studies. As far as cellular interaction was concerned, CS-PCL was the most efficient formulation for uptake of fluorescent markers Nile Red and Rhodamine123 incorporated into nanoparticles. Especially, CS-PCL nanoparticles loaded with Rhodamine123 sharing hydrophilic properties with MMC were selectively incorporated by bladder cancer cell line, but not by normal bladder epithelial cells. CS-PCL nanoparticles seem to be promising for MMC delivery with respect to anticancer efficacy tested against MB49 bladder carcinoma cell line. PMID- 19135515 TI - Voltammetric sensor for barbituric acid based on a sol-gel derivated molecularly imprinted polymer brush grafted to graphite electrode. AB - A voltammetric sensor based on a molecularly imprinted polymer (MIP) brush grafted to sol-gel film on graphite electrode is reported for the selective and sensitive analysis of barbituric acid (BA) in aqueous, blood plasma, and urine samples. The modified electrode was preanodised at +1.6 V (vs. saturated calomel electrode), where encapsulated BA involved hydrophobically induced hydrogen bondings, in MIP cavities exposed at the film/solution interface, at pH 7.0. Scanning electron microscopy (SEM) was employed to characterise the surface morphology of the resultant imprinted film of MIP brush. The differential pulse, cathodic stripping voltammetry (DPCSV) technique was employed to investigate the binding performance of the sol-gel-modified imprinted polymer brush, which yielded a linear response in the range of 4.95-100.00 microg mL(-1) of BA with a detection limit of 1.6 microg mL(-1) (S/N=3). PMID- 19135516 TI - Topical delivery of lipophilic drugs from o/w Pickering emulsions. AB - Surfactant-free emulsions stabilized by solid particles (Pickering emulsions) have been evaluated in the terms of skin absorption of lipophilic drugs. The behavior of three formulations: a surfactant-based emulsion, a Pickering emulsion stabilized by silica particles and a solution in triglyceride oil, were compared in order to assess the effect of the surface coating of Pickering emulsions as new dosage forms for topical application. Such comparative investigation was performed in vitro on excised pig skin in Franz diffusion cells with all-trans retinol as model lipophilic drug. Surfactant-based (classical, CE) and Pickering (PE) oil-in-water emulsions containing retinol were prepared with the same chemical composition (except the stabilizing agent: surfactant or silica particles), the same droplet size and the same viscosity. No permeation through the skin sample was observed after 24h exposure because of the high lipophilic character of retinol. Penetration of retinol was 5-fold larger for both CE and PE than for the solution in triglyceride. The distribution of retinol inside the skin layers depended significantly on the emulsions type: the classical emulsion allowed easy diffusion through the stratum corneum, so that large amounts reached the viable epidermis and dermis. Conversely, high storage of retinol inside the stratum corneum was favored by the Pickering emulsion. The retinol content in stratum corneum evaluated by skin stripping, demonstrated the increased retinol accumulation from PE. Therefore Pickering emulsions are new drug penetration vehicles with specific behavior; they are well-suited either for targeting the stratum corneum or aimed at slow release of drug from stratum corneum used as a reservoir to the deeper layers of skin. PMID- 19135517 TI - Sanguinarine induces apoptosis in A549 human lung cancer cells primarily via cellular glutathione depletion. AB - Sanguinarine is a plant-derived benzophenanthridine alkaloid and has been shown to possess anti-tumor activities against various cancer cells. In this study, we investigated whether sanguinarine induces apoptosis in A549 human lung cancer cells. Treatment of A549 cells with sanguinarine induced apoptosis in a dose- and time-dependent manner. Treatment with sanguinarine led to activation of caspases and MAPKs as well as increased MKP-1 expression. Importantly, pretreatment with z VAD-fmk, a pan caspase inhibitor suppressed the sanguinarine-induced apoptosis in A549 cells. Moreover, pretreatment with NAC, a sulfhydryl group-containing reducing agent strongly suppressed the apoptotic response and caspase activation to sanguinarine. However, the sanguinarine-mediated cytotoxicity in A549 cells was not protected by pharmacological inhibition of MAPKs or MKP-1 siRNA-mediated knockdown of MKP-1. These results collectively suggest that sanguinarine induces apoptosis in A549 cells through cellular glutathione depletion and the subsequent caspase activation. PMID- 19135518 TI - Plant derived antioxidants - Geraniol and camphene protect rat alveolar macrophages against t-BHP induced oxidative stress. AB - Exploration of antioxidants of plant origin and scientific validation of their efficacies has unraveled bioactives from natural sources. In this study, two terpenoids camphene and geraniol were assessed for their cytoprotective and antioxidant potential using t-BHP stressed rat alveolar macrophages. Effect of these test substances along with a known plant derived antioxidant quercetin was seen on cell viability, some oxidative stress markers as well as on mitochondrial membrane potential. Both the test substances geraniol and camphene increased the cell viability significantly as indicated by MTT assay and LDH release assay, during pre-treatment of test compound. Camphene and geraniol showed 29% (P<0.05) and 45% (P<0.05) increase in SOD activity, 28% and 120% (P<0.001) increase in GSH content and restored the mitochondrial membrane potential during pre-treatment as compared to stressed cells. Camphene and geraniol were found to significantly decrease lipid peroxidation, inhibit NO release (83.84% and 64.61%) and ROS generation in the pre-treated cells as compared to stressed cells. The test compounds also showed significant protection against ROS during post-treatment of the test compounds. Results indicate the pharmacological potential of these phytochemicals in lung inflammatory diseases where oxidative stress is a critical control point. PMID- 19135519 TI - Neuronal nitric oxide synthase in the olfactory system, forebrain, pituitary and retina of the adult teleost Clarias batrachus. AB - Immunocytochemical application of antibodies against nNOS to the brain sections of Clarias batrachus revealed intense immunoreactivity in several olfactory receptor neurons (ORNs), in their axons over the olfactory nerve, and terminals in the olfactory glomeruli. Several basal cells in the olfactory epithelium showed NOS immunoreactivity. Application of post-embedding immunoelectron microscopy showed nNOS labeled gold particles in apical cilia, dendrites and soma of the ORNs and also in the axon terminals in the glomeruli of the olfactory bulb. nNOS containing fibers were also encountered in the medial olfactory tracts (MOTs). Bilateral ablation of the olfactory organ resulted in total loss of nNOS immunoreactivity in the fascicles of the olfactory nerve layer and also in the MOT. nNOS immunoreactivity was seen in several cells of the nucleus preopticus (NPO) and their axons that innervate the pituitary gland. Some cells in the floor of the tuberal area were stained positive with nNOS antibodies. nNOS immunolabeled cells were seen in all the three components of the pituitary gland with light as well as post-embedding immunoelectron microscopy. While several nNOS immunoreactive fibers were seen in rostral pars distalis, a much limited fiber population was seen in the proximal pars distalis. In addition, conspicuous immunoreactivity was noticed in some ganglion cells in the retina and in some fibers of the optic nerve traceable to the optic tectum. The NO containing system in this fish appears to be similar to that in other fishes. PMID- 19135520 TI - Antioxidant activity of blueberry fruit is impaired by association with milk. AB - The antioxidant properties of dietary phenolics are believed to be reduced in vivo because of their affinity for proteins. In this study we assessed the bioavailability of phenolics and the in vivo plasma antioxidant capacity after the consumption of blueberries (Vaccinium corymbosum L.) with and without milk. In a crossover design, 11 healthy human volunteers consumed either (a) 200 g of blueberries plus 200 ml of water or (b) 200 g of blueberries plus 200 ml of whole milk. Venous samples were collected at baseline and at 1, 2, and 5 h postconsumption. Ingestion of blueberries increased plasma levels of reducing and chain-breaking potential (+6.1%, p<0.001; +11.1%, p<0.05) and enhanced plasma concentrations of caffeic and ferulic acid. When blueberries and milk were ingested there was no increase in plasma antioxidant capacity. There was a reduction in the peak plasma concentrations of caffeic and ferulic acid (-49.7%, p<0.001, and -19.8%, p<0.05, respectively) as well as the overall absorption (AUC) of caffeic acid (p<0.001). The ingestion of blueberries in association with milk, thus, impairs the in vivo antioxidant properties of blueberries and reduces the absorption of caffeic acid. PMID- 19135521 TI - Influence of polyphenols on Escherichia coli resistance to oxidative stress. AB - Among all polyphenols tested (tannic acid and flavonoids belonging to different subclasses) only tannin and quercetin significantly enhanced resistance of Escherichia coli to peroxide stress. Pretreatment of the cells with quercetin and tannin resulted in a decrease in the growth arrest duration under moderate H(2)O(2) concentration (2 mM) and an increase in survival under high (10 mM) doses. The shorter growth recovery period in pretreated cells was connected with more rapid H(2)O(2) elimination because of induced activity of scavenging enzymes. This effect was absent in the Delta oxyR mutant, which was unable to induce genes responding to peroxide stress. The data obtained suggest that the observed protection was a result of two overlapping effects: induction of OxyR regulon by low concentrations of H(2)O(2), accumulated during extracellular autoxidation of quercetin and tannin, and protection of synthesis of OxyR regulated antioxidant enzymes during H(2)O(2) stress because of intracellular binding of iron by quercetin and tannin and suppressing Fenton chemistry. PMID- 19135522 TI - Oxidative stress in rats fed a high-fat high-sucrose diet and preventive effect of polyphenols: Involvement of mitochondrial and NAD(P)H oxidase systems. AB - Mitochondrial and NADPH oxidase systems and oxidative stress were investigated in 12 week high-fat high-sucrose (HFHS) diet-fed rats. A protective effect of wine polyphenol (PP) extract was also examined. In liver, maximal activities of CII and CII+III mitochondrial complexes were decreased but NADPH oxidase expression (p22(phox) and p47(phox)) and NADPH oxidase-dependent superoxide anion production were not modified, whereas oxidative stress (lipid and protein oxidation products and antioxidant systems) was increased with HFHS diet. In muscle, anion superoxide production was slightly increased while mitochondrial complex activities and lipid and protein oxidation products were not modified with HFHS diet. In heart, NADPH oxidase expression and superoxide anion production were increased, and maximal activity of mitochondrial respiratory chain complexes or oxidative stress parameters were not modified. Wine polyphenol extract had an inhibiting effect on liver oxidative stress and on heart NADPH oxidase expression and superoxide anion production, and on induction of hepatic steatosis with HFHS diet. Induction of mitochondrial dysfunction could be a primary event in the development of oxidative stress in liver, while in skeletal muscle and in heart the NADPH oxidase system seems to be mainly involved in oxidative stress. Wine polyphenol extract was shown to partially prevent oxidative stress in liver and heart tissues and to nearly completely prevent steatosis development in liver. PMID- 19135523 TI - Hypothermic preconditioning of endothelial cells attenuates cold-induced injury by a ferritin-dependent process. AB - Hypothermia for myocardial protection or storage of vascular grafts may damage the endothelium and impair vascular function upon reperfusion/rewarming. Catalytic iron pools and oxidative stress are important mediators of cold-induced endothelial injury. Because endothelial cells are highly adaptive, we hypothesized that hypothermic preconditioning (HPC) protects cells at 0 degrees C by a heme oxygenase-1 (HO-1) and ferritin-dependent mechanism. Storage of human coronary artery endothelial cells at 0 degrees C caused the release of lactate dehydrogenase, increases in bleomycin-detectible iron (BDI), and increases in the ratio of oxidized/reduced glutathione, signifying oxidative stress. Hypoxia increased injury at 0 degrees C but did not increase BDI or oxidative stress further. HPC at 25 degrees C for 15-72 h attenuated these changes by an amount achievable by pretreating cells with 10-20 microM deferoxamine, an iron chelator, and protected cell viability. Treating cells with hemin chloride at 37 degrees C transiently increased intracellular heme, HO-1, BDI, and ferritin. Elevated heme/iron sensitized cells to 0 degrees C but ferritin was protective. HPC increased iron maximally after 2 h at 25 degrees C and ferritin levels peaked after 15 h. HO-1 was not induced. When HPC-mediated increases in ferritin were blocked by deferoxamine, protection at 0 degrees C was diminished. We conclude that HPC-mediated endothelial protection from hypothermic injury is an iron- and ferritin-dependent process. PMID- 19135525 TI - Attenuation of lipopolysaccharide-induced oxidative stress and apoptosis in fetal pulmonary artery endothelial cells by hypoxia. AB - Pulmonary vascular endothelial injury resulting from lipopolysaccharide (LPS) and oxygen toxicity contributes to vascular simplification seen in the lungs of premature infants with bronchopulmonary dysplasia. Whether the severity of endotoxin-induced endothelial injury is modulated by ambient oxygen tension (hypoxic intrauterine environment vs. hyperoxic postnatal environment) remains unknown. We posited that ovine fetal pulmonary artery endothelial cells (FPAEC) will be more resistant to LPS toxicity under hypoxic conditions (20-25 Torr) mimicking the fetal milieu. LPS (10 microg/ml) inhibited FPAEC proliferation and induced apoptosis under normoxic conditions (21% O(2)) in vitro. LPS-induced FPAEC apoptosis was attenuated in hypoxia (5% O(2)) and exacerbated by hyperoxia (55% O(2)). LPS increased intracellular superoxide formation, as measured by 2 hydroxyethidium (2-HE) formation, in FPAEC in normoxia and hypoxia. 2-HE formation in LPS-treated FPAEC increased in parallel with the severity of LPS induced apoptosis in FPAEC, increasing from hypoxia to normoxia to hyperoxia. Differences in LPS-induced apoptosis between hypoxia and normoxia were abolished when LPS-treated FPAEC incubated in hypoxia were pretreated with menadione to increase superoxide production. Apocynin decreased 2-HE formation, and attenuated LPS-induced FPAEC apoptosis under normoxic conditions. We conclude that ambient oxygen concentration modulates the severity of LPS-mediated injury in FPAEC by regulating superoxide levels produced in response to LPS. PMID- 19135524 TI - Esterified trityl radicals as intracellular oxygen probes. AB - Triarylmethyl (trityl) radicals exhibit high stability and narrow linewidth under physiological conditions which provide high sensitivity and resolution for the measurement of O2 concentrations, making them attractive as EPR oximetry probes. However, the application of previously available compounds has been limited by their poor intracellular permeability. We recently reported the synthesis and characterization of esterified trityl radicals as potential intracellular EPR probes and their oxygen sensitivity, redox properties, and enzyme-mediated hydrolysis were investigated. In this paper, we report the cellular permeability and stability of these trityls in the presence of bovine aortic endothelial cells. Results show that the acetoxymethoxycarbonyl-containing trityl AMT-02 exhibits high stability in the presence of cells and can be effectively internalized. The intracellular hydrolysis of AMT-02 to the carboxylate form of the trityl (CT-03) was also observed. In addition, this internalized trityl probe was applied to measure intracellular O2 concentrations and the effects of menadione and KCN on the rates of O2 consumption in endothelial cells. This study demonstrates that these esterified trityl radicals can function as effective EPR oximetry probes measuring intracellular O2 concentration and consumption. PMID- 19135526 TI - Fluorescent detection of alpha-aminoadipic and gamma-glutamic semialdehydes in oxidized proteins. AB - The oxidative modification of proteins is believed to play a critical role in the etiology and/or progression of several diseases. alpha-Aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS) residues represent major oxidized amino acids generated in oxidized proteins. This paper describes a novel procedure for the specific and sensitive determination of AAS and GGS after their reductive amination with sodium cyanoborohydride and p-aminobenzoic acid, a fluorescence reagent, to their corresponding derivatives, followed by a high performance liquid chromatography (HPLC) analysis. This fluorescent labeling of protein-associated aldehyde moieties is a simple and accurate technique that may be widely used to reveal increased levels of oxidatively modified proteins with reactive oxygen species during aging and disease. PMID- 19135527 TI - Pasteurella multocida toxin activates Gbetagamma dimers of heterotrimeric G proteins. AB - The mitogenic Pasteurella multocida toxin (PMT) is a major virulence factor of P. multocida, which causes Pasteurellosis in man and animals. The toxin activates the small GTPase RhoA, the MAP kinase ERK and STAT proteins via the stimulation of members of two G protein families, G(q) and G(12/13). PMT action also results in an increase in inositol phosphates, which is due to the stimulation of PLCbeta via Galpha(q). Recent studies indicate that PMT additionally activates Galpha(i) to inhibit adenylyl cyclase. Here we show that PMT acts not only via Galpha but also through Gbetagamma signaling. Activation of Gbetagamma by PMT causes stimulation of phosphoinositide 3-kinase (PI3K) gamma and formation of phosphatidylinositol-3,4,5-trisphosphate (PIP(3)) as indicated by the recruitment of a PIP(3)-binding pleckstrin homology (PH) domain-containing protein to the plasma membrane. Moreover, it is demonstrated that Gbetagamma is necessary for PMT-induced signaling via Galpha. Mutants of Galpha(q) incapable of binding or releasing Gbetagamma are not activated by PMT. Similarly, sequestration of Gbetagamma inhibits PMT-induced Galpha-signaling. PMID- 19135528 TI - Intracellular trafficking and assembly of specific Kir3 channel/G protein complexes. AB - We have previously demonstrated that Kir3.1 channels and Gbeta1gamma2 subunits initially interact in the endoplasmic reticulum (ER). To elucidate the role that anterograde protein trafficking pathways may play in the formation of these complexes, we used dominant negative (DN) mutants of the small G proteins Sar 1 and various compartment-specific Rabs which impede anterograde protein trafficking at different steps. Sar 1 H79G and Rab 1 S25N mutants efficiently blocked the plasma membrane trafficking of the Kir3.1/Kir3.4 complex however they did not block the Gbeta1gamma2/Kir3.1 interaction as measured using bioluminescence resonance energy transfer (BRET). This interaction was also insensitive to the presence of DN Rabs 2, 6, 8, and 11. These results confirm that Gbetagamma/Kir3 complexes form early during channel biosynthesis and trafficking. Using a combination of BRET, protein complementation assays and co immunoprecipitation, we demonstrate that Gbeta1-4 can interact with Kir3.1 in the absence of Kir3.4. Gbeta5 does not directly interact with the channel but can still be co-immunoprecipitated as part of a larger complex. The interaction between Gbeta and Kir3.1 was selectively fostered by co-expression with different Ggamma subunits. When Ggamma1 or Ggamma11 was co-expressed with eGFP-Gbeta3 or eGFP-Gbeta4, the interaction with the effector was lost. Kir3.2 was capable of interacting with Gbeta1-3 and not Gbeta4 or Gbeta5. These interactions were again fostered by co-expression with Ggamma and were also insensitive to DN Sar 1 or Rab 1. Taken together, our data show that these "precocious" channel/G protein interactions are specific and may have implications beyond their basic function in signalling events. PMID- 19135529 TI - Presence of duplicate genes encoding a phylogenetically new subgroup of form I ribulose 1,5-bisphosphate carboxylase/oxygenase in Mycobacterium sp. strain JC1 DSM 3803. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the key enzyme of the Calvin reductive pentose phosphate cycle. Two sets of the structural genes for form I RubisCO were identified in Mycobacterium sp. strain JC1. The genes were clustered on the chromosome in the transcriptional order of cbbL-cbbS. Cloned cbbL-1 and cbbS-1 and cbbL-2 and cbbS-2 have open reading frames of 1431, 426, 1428, and 426 nucleotides, respectively. Primer extension analysis revealed that transcriptional start sites of cbbLS-1 and -2 genes were the nucleotides T and G located 99 and 41bp upstream of the cbbL start codons, respectively. CbbLS 1 and CbbLS-2 that were expressed in Escherichia coli exhibited RubisCO activity. A phylogeny of CbbL amino acid sequences revealed that the two enzymes in Mycobacterium sp. strain JC1 may form a new phylogenetic subgroup, type IE, in the 'red-like' group of the form I RubisCO family. PMID- 19135530 TI - Effect of P-glycoprotein inhibitor, verapamil, on oral bioavailability and pharmacokinetics of irinotecan in rats. AB - The objective of present investigation was to study the effect of verapamil on the pharmacokinetics of irinotecan in order to evaluate the role of P glycoprotein (P-gp) in irinotecan disposition. An in vitro study using Caco-2 intestinal cell monolayer was first carried out to determine the effect of verapamil on the function of intestinal P-gp. Verapamil (25mg/kg) was administered orally 2h before irinotecan oral (80 mg/kg) or intravenous (20mg/kg) dosing in female Wistar rats. Plasma and biliary samples were collected at specified time points from control and treated animals to determine irinotecan and its metabolite, SN-38 concentrations. Bi-directional transport and inhibition studies in Caco-2 cells indicated irinotecan to be a P-gp substrate and the function of intestinal P-gp was significantly inhibited in presence of verapamil. After oral irinotecan dosing, the mean area under the plasma concentration-time curve (AUC) was found to be 14.03+/-2.18 microgh/ml which was increased significantly, i.e. 61.71+/-15.0 microgh/ml when verapamil was co-administered (P<0.05). Similarly, the mean maximum plasma concentration of irinotecan increased from 2.93+/-0.37 microg/ml (without verapamil) to 10.75+/-1.0 microg/ml (with verapamil) (P<0.05). There was approximately 4-5-folds increase in apparent bioavailability. On the other hand, the intravenous irinotecan administration with verapamil resulted in small but statistically significant effect on AUC (10.76+/-2.0 to 23.3+/-3.8 microgh/ml; P<0.05) and systemic clearance (1206.4+/ 159.7 to 713.5+/-78.2 ml/(hkg)). In addition, SN-38 showed significant change in oral pharmacokinetic parameters and minor changes in intravenous pharmacokinetic profile. Biliary excretion curves of both irinotecan and SN-38 were lowered by verapamil. The mean percent of irinotecan excreted into bile over 5h following intravenous and oral administration was found to be 8% and 1%, respectively, which was further reduced to half when treated with verapamil. These results are quite stimulating for further development of a clinically useful oral formulation of irinotecan based on P-gp inhibition. PMID- 19135531 TI - Dry powder inhalations containing thymopentin and its immunomodulating effects in Wistar rats. AB - Thymopentin (TP5), a synthetic pentapeptide, has been used in clinic as a modulator for immunodeficiences through intramuscular administration. The purpose of this study was to design and evaluate dry powder inhalations (DPIs) for pulmonary delivery of TP5. Dry powder inhalations containing leucine (a dispersibility enhancer), mannitol, and lactose (bulking agents) were prepared by spray-drying from aqueous formulations. The formulation components on the aerosolisation characteristics of spray-dried powders were investigated through the use of various amount of leucine, lactose and mannitol. Following spray drying, resultant powders were characterized using scanning electron microscopy, laser diffraction and tapped density measurements, and the aerosolisation performance was determined using Twin Stage Impinger. The immunosuppression Wistar rats model was constructed to evaluate the immunomodulating effects of TP5 DPIs in vivo. The results of T-lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+ ratio) analyses suggest that TP5 DPIs have modulating effects. On an overall evaluation, TP5 pulmonary delivery DPIs may be feasible for the future clinical application. PMID- 19135532 TI - Modulation of release rate and barrier transport of Diclofenac incorporated in hydrophilic matrices: role of cyclodextrins and implications in oral drug delivery. AB - The aim of this work was to investigate how the incorporation of a hydrophilic cyclodextrin (CD) inside erodible hydrophilic matrices affects drug-release behavior and transport properties through artificial and biological membranes. To this purpose, Diclofenac (Dic) was incorporated in poly(ethyleneoxide) (PEO) matrices as poorly soluble free acid (DicH) or freely water-soluble sodium salt (DicNa) in the presence or absence of hydroxypropyl-beta-cyclodextrin (HP beta CD). Preliminary experiments demonstrated that HP beta CD increased Dic apparent solubility as a function of its amount in the solution and medium pH due to complex formation. Permeation of ionized Dic through porcine buccal mucosa gave higher values of J(SS) and K(p) as compared to silicon membranes and depended on the presence of HP beta CD. Incorporation of HP beta CD in PEO tablets resulted in an increase of release rate for both forms of Dic whereas cumulative drug flux through silicon membranes and porcine buccal mucosa was increased for DicH and decreased for DicNa. An interpretation of this behavior was attempted on the basis of the presence of a transport resistance occurring inside the hydrated gel matrix as modified by the presence of CD. In conclusion, this study has demonstrated that the use of CDs in hydrophilic matrices intended for oral drug delivery should be rationalized since their modulator effect relies not only on drug-dissolution rate but also on environment where drug release occurs (aqueous medium, membrane interface). PMID- 19135533 TI - Intraspinal cord delivery of IGF-I mediated by adeno-associated virus 2 is neuroprotective in a rat model of familial ALS. AB - BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a devastating disease that is characterized by the progressive loss of motor neurons. Patients with ALS usually die from respiratory failure due to respiratory muscle paralysis. Consequently, therapies aimed at preserving segmental function of the respiratory motor neurons could extend life for these patients. Insulin-like growth factor-I (IGF-I) is known to be a potent survival factor for motor neurons. In this study we induced high levels of IGF-I expression in the cervical spinal cord of hSOD1(G93A) rats with intraspinal cord (ISC) injections of an adeno-associated virus serotype 2 vector (CERE-130). This approach reduced the extent of motor neuron loss in the treated segments of the spinal cord. However, a corresponding preservation of motor function was observed in male, but not female, hSOD1(G93A) rats. We conclude that ISC injection of CERE-130 has the potential to protect motor neurons and preserve neuromuscular function in ALS. PMID- 19135534 TI - Novel dimerization mode of the human Bcl-2 family protein Bak, a mitochondrial apoptosis regulator. AB - Interactions of Bcl-2 family proteins play a regulatory role in mitochondrial apoptosis. The pro-apoptotic protein Bak resides in the outer mitochondrial membrane, and the formation of Bak homo- or heterodimers is involved in the regulation of apoptosis. The previously reported structure of the human Bak protein (residues Glu16-Gly186) revealed that a zinc ion was coordinated with two pairs of Asp160 and His164 residues from the symmetry-related molecules. This zinc-dependent homodimer was regarded as an anti-apoptotic dimer. In the present study, we determined the crystal structure of the human Bak residues Ser23-Asn185 at 2.5A, and found a distinct type of homodimerization through Cys166 disulfide bridging between the symmetry-related molecules. In the two modes of homodimerization, the molecular interfaces are completely different. In the membrane-targeted model of the S-S bridged dimer, the BH3 motifs are too close to the membrane to interact directly with the anti-apoptotic relatives, such as Bcl x(L). Therefore, the Bak dimer structure reported here may represent a pro apoptotic mode under oxidized conditions. PMID- 19135535 TI - Using fossils and molecular data to reveal the origins of the Cape proteas (subfamily Proteoideae). AB - The angiosperm family Proteaceae is a distinct component of the Cape Floristic Region biodiversity hotspot with 330 endemic species. Phylogenetic analyses of subfamily Proteoideae using sequence data from one nuclear and six plastid loci show that most of this diversity is contained in two distinct Cape floral clades. Molecular dating analyses, using Bayesian and penalized likelihood methods and four phylogenetically supported fossil age constraints, reveal contrasting histories for these two clades. The genus Protea belongs to a lineage that may have been in Africa since the Late Cretaceous but began to diversify in the Cape only 5-18 Myr ago. In contrast, the Leucadendrinae clade presumably arrived in the region no earlier than 46 Myr ago by long-distance dispersal from an Australian ancestor and the extant members of this clade began to diversify in the Cape 22-39 Myr ago. These results join a growing number of case studies that challenge the commonly accepted view that most of the Cape flora radiated synchronously in the Late Miocene and Early Pliocene when a Mediterranean climate settled in the region. PMID- 19135536 TI - Retropositional events consolidate the branching order among New World monkey genera. AB - Due to contradicting relationships obtained from various morphological and genetic studies, phylogenetic relationships among New World monkey genera are highly disputed. In the present study, we analyzed the presence/absence pattern of 128 SINE integrations in all New World monkey genera. Among them, 70 were specific for only a single genus, whereas another 18 were present in all New World monkey genera. The 40 remaining insertions were informative to elucidate phylogenetic relationships among genera. Several of them confirmed the monophyly of the three families Cebidae, Atelidae and Pitheciidae as well as of the subfamily Callithrichinae. Further markers provided evidence for a sister grouping of Cebidae and Atelidae to the exclusion of Pitheciidae as well as for relationships among genera belonging to Callithrichinae and Atelidae. Although a close affiliation of Saimiri, Aotus and Cebus to Callithrichinae was shown, the relationships among the three genera remained unresolved due to three contradicting insertions. PMID- 19135537 TI - Calculation of QT shift in non clinical safety pharmacology studies. AB - INTRODUCTION: Drug-induced QT interval prolongation is a major concern in new drug candidate development. This study presents a method of assessment of drug induced QT interval prolongation without need for QT correction in conscious Beagle dogs and Cynomolgus monkeys monitored by telemetry. Accuracy and reliability are analysed by comparison with a reference QT correction method (Holzgrefe) from experiments performed with reference substances terfenadine, thioridazine and sotalol. METHODS: The QT shift method principle is assessment of any drug-induced QT interval shift directly from the individual QT/RR relationship. The individual QT/RR relationship is built from a treatment-free 24 hour recording period. QT and RR intervals are determined from a beat-to-beat analysis. A probabilistic method is used to define the individual QT/RR relationships. Checks were performed to compare results obtained with the QT shift method and the QT correction methods. The robustness of the QT shift method was tested under various conditions of drug-induced heart rate change (i.e. normal, bradycardia and tachycardia). RESULTS: The extent of agreement with the used reference QT correction method, Holzgrefe formula, was excellent (3-4 ms) in both animal species under the various drug induced effects on heart rate. The statistical sensitivity threshold for detection of QT prolongation according to a standard safety pharmacology study design was 7-8 ms. DISCUSSION: When combined with the probabilistic determination of individual QT/RR relationships, this simple method provides a direct assessment of a drug-induced effect on QT interval, without any curve fitting or application of correction formula. Despite noticeably different shapes in QT/RR relationships, the QT shift method is applicable to both Beagle dogs and Cynomolgus monkeys. It is likely that the QT shift method will be particularly helpful in problematic cases, enabling detection of drug-induced prolongation of less than 10 ms. PMID- 19135538 TI - A high molecular weight polypeptide cross-reacting with the antibodies to the dynein heavy chain localizes to the subset of Golgi complex in higher plant cells. AB - Antibodies were produced against fragments of the microtubule-binding domain and the motor domain of the dynein heavy chain from Dictyostelium discoideum to probe whole cell extracts of root meristem cells of wheat Triticum aestivum. In plant extracts, these antibodies cross-reacted with a polypeptide of high molecular weight (>500kDa). The antibodies bound to protein A-Sepharose precipitated high molecular weight polypeptide from cell extracts. Immunofluorescence showed that the antibodies identified various aggregates inside cells, localized at the perinuclear area during interphase to early prophase, at the spindle periphery and polar area during mitosis, and in the interzonal region during phragmoplast development. Some aggregates were also co-labeled by markers for the Golgi apparatus. Thus, we found in higher plant cells a high molecular weight antigen cross-reacting with the antibodies to motor and microtubule-binding domains of dynein heavy chains. This antigen is associated with aggregates distributed in the cytoplasm in cell cycle-dependent manner. A subset of these aggregates belongs to the Golgi complex. PMID- 19135539 TI - Acute systemic fibroblast growth factor-2 enhances long-term memory in developing rats. AB - Recent findings have provided much insight into the mechanisms underlying long term memory formation, and it is now known that long-term memory depends on the activation of a molecular cascade that culminates with structural changes in the brain. However, little is known about the signals that give rise to or regulate these structural changes. In this article we propose that fibroblast growth factor-2 (FGF2), a mitogen for several cell types, may be one of the molecular signals critically involved in the structural changes underlying long-term memory. If FGF2 is part of the signalling cascade involved in long-term memory, then increasing the activation of FGF2 should facilitate memory. In Experiments 1 and 2, we demonstrated that systemic injection of FGF2 (20ng/g of body weight) facilitated memory for contextual fear in 16, 19, and 22 day old male Sprague Dawley rats. Experiment 3 demonstrated that the observed facilitation of memory was not due to FGF2 increasing rats' sensitivity to footshock. These results implicate FGF2 as a possible molecular signal in long-term memory, and further, illustrate a novel means of enhancing memory. PMID- 19135541 TI - Prenatal environmental influences on the production of sex-specific traits in mammals. AB - The determination of offspring sex represents a delicate balancing act during which offspring must pass through several developmental levels in the presence of influential environmental factors. Successful expression of sex-specific traits requires genetic and hormone-based effects on several organizational and activational levels. Environmental factors can exert controls and disruptions at each of these levels. This review addresses the developmental stages at which environmental factors may influence the processes of sex determination, with an in depth focus on the prenatal stages, including the production of the primary and secondary sex ratios and the differentiation of functional secondary sexual characters. PMID- 19135540 TI - Clostridium taeniosporum is a close relative of the Clostridium botulinum Group II. AB - Clostridium taeniosporum is a Gram-positive, anaerobic, rod-shaped non-toxigenic organism isolated from Crimean lake silt. It is unique in forming spores from which about twelve large, flat, ribbon-like appendages emanate. These ribbon-like structures, about 4.5 microm long and 0.45 microm wide, are assembled from smaller fibrils with 5 nm diameter spherical heads attached to thin tails about 1 2 nm in diameter and about 40 nm in length. The appendages have four major components, a glycoprotein with a collagen-like region, two proteins each of which contains two conserved domains of unknown function, and an ortholog of the Bacillus subtilis spore morphogenetic protein SpoVM. Genes for three of these and other, possibly related proteins, cluster on two chromosome fragments. Here we report that C. taeniosporum is saccharolytic, non-proteolytic, and produces both acetic and butyric acid fermentation products. It synthesizes alpha-D-glucosidase and N-acetyl-beta,D-glucoseaminidase constitutively. These physiological properties are similar to those of the C. botulinum Group II. Genotypically, C. taeniosporum is also closely related to the same Group II, based on 16S rDNA sequences. C. taeniosporum differs from typical C. botulinum Group II strains because it is non-toxigenic and in forming the ribbon-like spore appendages. These major differences among otherwise closely related organisms suggest lateral transfer of genes for appendage synthesis and for toxigenicity. PMID- 19135542 TI - Pivotal role of c-Fos in nitric oxide synthase 2 expression in airway epithelial cells. AB - The regulation of nitric oxide synthase 2 (NOS2) in airway epithelial cells plays a key role in the innate host response to a wide variety of microbial agents and also participates in the generation of pathologic airway inflammation. Among the important signalling cascades that direct NOS2 gene expression are nuclear factor kappaB (NFkappaB) and interferon-gamma (IFNgamma)/signal transducer and activator of transcription 1 (STAT-1). Previous studies suggest activator protein-1 (AP-1), in particular c-Fos component of AP-1, influences NOS2 expression. We investigated the effect of c-Fos modulation using RNA interference siRNA on NOS2 gene expression. A549 cells stably transfected with a plasmid overexpressing a c Fos siRNA construct (FOSi) resulted in a decrease of NOS2 protein inducibility by IFN gamma. In contrast, classical IFN gamma inducible signal transduction pathways interferon regulated factor-1 (IRF-1) and pSTAT-1 were activated at a similar magnitude in FOSi and control cells. DNA-protein binding assays showed that c-Fos binding was present in wild type cells, but reduced in FOSi clones. FOSi clones had activation of NFkappaB detectable by DNA-protein binding assays, which may have contributed to a decrease of NOS2 expression. Overall, these studies indicate that c-Fos is a requisite and specific component for inducible NOS2 expression. PMID- 19135543 TI - Workshop--cough: exercise, speech and music. AB - Twelve distinguished scientists attended the workshop, heard three presentations, and took part in the discussions. Fontana first described his unpublished studies on cough in exercise and during hyperventilation with healthy subjects. Both activities depressed cough induced by inhalation of distilled water aerosol (fog). The possible mechanisms were discussed. Gibson then described the successful use of speech therapy to treat chronic cough, and discussed the possible mechanisms, centering on the role of the larynx and its neural control. A comparison was made with the ability of speech and laughter to precipitate cough. Widdicombe discussed the scanty literature on the effect of singing and playing wind instruments on cough, most of the evidence being anecdotal. In the discussion periods several matters for future study arose. It is usually not clear if the modulation of cough, its depression, enhancement or excitation, arose primarily at peripheral sites (reflexes from the airways), or at a cortical level, or both. Nor is it clear whether the same results would be obtained with provoked cough and with spontaneous cough. But all three aspects of 'behavioual' changes in cough sensitivity (exercise, speech and music) could be further explored, and current techniques should make this possible. PMID- 19135544 TI - More efficient mastication allows increasing intake without compromising digestibility or necessitating a larger gut: comparative feeding trials in banteng (Bos javanicus) and pygmy hippopotamus (Hexaprotodon liberiensis). AB - The digestion of plant material in mammalian herbivores basically depends on the chemical and structural composition of the diet, the mean particle size to which the forage is processed, and the ingesta retention time. These different factors can be influenced by the animal, and they can presumably compensate for each other. The pygmy hippopotamus, a non-ruminating foregut fermenter, has longer mean retention times than ruminants; however hippos do not achieve higher (fibre) digestibilities on comparable diets, which could be due to ineffective mastication. We performed feeding trials with six pygmy hippos (Hexaprotodon liberiensis) and six banteng cattle (Bos javanicus) on a grass diet. As predicted, both species achieved similar dry matter, organic matter, crude protein and gross energy digestibilities. However, neutral and acid detergent fibre digestibility was lower in pygmy hippos. Apparently, in these species, fibre digestibility was more influenced by particle size, which was larger in pygmy hippos compared to banteng, than by retention time. In spite of their higher relative food intake, the banteng in this study did not have greater relative gut fills than the hippos. Ruminants traditionally appear intake-limited when compared to equids, because feed particles above a certain size cannot leave the rumen. But when compared to nonruminating foregut fermenters, rumination seems to free foregut fermenters from an intrinsic food intake limitation. The higher energy intakes and metabolic rates in wild cattle compared to hippos could have life-history consequences, such as a higher relative reproductive rate. PMID- 19135545 TI - Lipid, fatty acid and protein content of late larval to early juvenile stages of the western rock lobster, Panulirus cygnus. AB - Lipid, fatty acid and protein content were determined individually on 7 phyllosomata, 69 clear pueruli, 286 pre-moult pueruli, and 86 juvenile western rock lobster (WRL) collected from four locations between the settlement seasons 2000 to 2006 to evaluate compositional changes during the non-feeding puerulus stage. Only the lipid content, particularly the phospholipids, decreased significantly with development. Protein declined sharply following moult to the juvenile. PL comprised between 86-94% of total lipid in all animals, and declined most between phyllosomata and clear pueruli (238.5 to 121.4 mg g(-1) DW) (p<0.001). Triacylglycerols were the only lipid to increase in absolute amounts with development, but declined 53% on average following moult to juvenile. This increase in TAG is likely due to the conversion of phospholipids to triacylglycerols. Monounsaturated fatty acids were the main energy form utilised during benthic development while polyunsaturated fatty acids showed a high degree of sparing. The n-3:n-6 fatty acid ratio of juveniles indicates that they may be approaching critically low levels of stored lipid energy reserves. Both protein, and lipid, declined sharply from the final puerulus phase to the juvenile confirming that a high energetic demand is required to fuel the moulting process. PMID- 19135546 TI - High-dose lovastatin decreased basal prostacyclin production in cultured endothelial cells. AB - The effect of lovastatin, a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, on prostacyclin production was studied in cultured human umbilical vein endothelial cells. The results indicated that lovastatin induced a significant dose- and time-dependent decrease of arachidonic acid release from the cells, an inhibition of phospholipase A(2) (PLA(2)) activity, a reduction of [(3)H]choline in lysophosphatidylcholine (lysoPC), and a diminishment of (45)Ca(2+) influx into the cells. The decreased arachidonic acid release was not reversed by addition of either intermediate products during cholesterol synthesis, such as mevalonate, geranylgeraniol, farnesol, or cholesterol and lipoprotein although a reduced concentration of cholesterol in the cells, caused by lovastatin, was reversed by added cholesterol. Lovastatin, furthermore, diminished prostacyclin production and inhibited activity of cyclooxygenase-1. 27 Hydroxycholesterol (27OHC), an oxidized cholesterol, had the same effect on HMG CoA reductase as lovastatin, but 27OHC increased arachidonic acid release and (45)Ca(2+) influx. Our results indicated that lovastatin inhibited significantly activities of PLA(2) and cyclooxygenase-1, resulting in a marked reduction in arachidonic acid release, lysoPC content and prostacyclin production in the cultured vascular endothelial cells. PMID- 19135547 TI - Functional study of cytoplasmic loops of human skeletal muscle chloride channel, hClC-1. AB - The membrane-resident domain of chloride channels and transporters of the CLC family is composed of 18 alpha-helices (designated A to R) connected sequentially by extracellular and intracellular loops, whose functional characteristics are generally unclear. To study the relevance of the intracellular loops linking helices D and E, F and G, H and I and J and K, alanine-exchange mutagenesis, split channel strategy, GST (glutathione transferase)-pull-down methods and whole cell patch-clamp recordings were used. We investigated the possible roles of these loops in binding to the cytoplasmic, carboxyl tail (C-tail) of the protein, as well as their physiological roles in channel function. Although other interacting positions are conceivable, our results indicate that there is unlikely to be significant binding between the C-tail and any one of these individual cytoplasmic loops. Particular loops might, however, be essential for some channel characteristics. For example, alanine-exchange mutation of the loop linking helix D to E eliminated channel currents; of the loop linking helix H to I caused a significant shift of the open probability of fast gating (P(o)(f)), towards more positive voltages; and of the loop linking helix J to K locked the common gating of hClC-1 open. Therefore, the gating mechanisms of hClC-1 might not only involve the helices and the C-tail, but also involve some of the loops. PMID- 19135548 TI - The Rho GTPase Wrch1 regulates osteoclast precursor adhesion and migration. AB - An excess of osteoclastic bone resorption relative to osteoblastic bone formation results in progressive bone loss, characteristic of osteoporosis. Understanding the mechanisms of osteoclast differentiation is essential to develop novel therapeutic approaches to prevent and treat osteoporosis. We showed previously that Wrch1/RhoU is the only RhoGTPase whose expression is induced by RANKL during osteoclastogenesis. It associates with podosomes and the suppression of Wrch1 in osteoclast precursors leads to defective multinucleated cell formation. Here we further explore the functions of this RhoGTPase in osteoclasts, using RAW264.7 cells and bone marrow macrophages as osteoclast precursors. Suppression of Wrch1 did not prevent induction of classical osteoclastic markers such as NFATc1, Src, TRAP (Tartrate-Resistant Acid Phosphatase) or cathepsin K. ATP6v0d2 and DC-STAMP, which are essential for fusion, were also expressed normally. Similar to the effect of RANKL, we observed that Wrch1 expression increased osteoclast precursor aggregation and reduced their adhesion onto vitronectin but not onto fibronectin. We further found that Wrch1 could bind integrin beta3 cytoplasmic domain and interfered with adhesion-induced Pyk2 and paxillin phosphorylation. Wrch1 also acted as an inhibitor of M-CSF-induced prefusion osteoclast migration. In mature osteoclasts, high Wrch1 activity inhibited podosome belt formation. Nevertheless, it had no effect on mineralized matrix resorption. Our observations suggest that during osteoclastogenesis, Wrch1 potentially acts through the modulation of alphav beta3 signaling to regulate osteoclast precursor adhesion and migration and allow fusion. As an essential actor of osteoclast differentiation, the atypical RhoGTPase Wrch1/RhoU could be an interesting target for the development of novel antiresorptive drugs. PMID- 19135549 TI - Target discovery from data mining approaches. AB - Data mining of available biomedical data and information has greatly boosted target discovery in the 'omics' era. Target discovery is the key step in the biomarker and drug discovery pipeline to diagnose and fight human diseases. In biomedical science, the 'target' is a broad concept ranging from molecular entities (such as genes, proteins and miRNAs) to biological phenomena (such as molecular functions, pathways and phenotypes). Within the context of biomedical science, data mining refers to a bioinformatics approach that combines biological concepts with computer tools or statistical methods that are mainly used to discover, select and prioritize targets. In response to the huge demand of data mining for target discovery in the 'omics' era, this review explicates various data mining approaches and their applications to target discovery with emphasis on text and microarray data analysis. Two emerging data mining approaches, chemogenomic data mining and proteomic data mining, are briefly introduced. Also discussed are the limitations of various data mining approaches found in the level of database integration, the quality of data annotation, sample heterogeneity and the performance of analytical and mining tools. Tentative strategies of integrating different data sources for target discovery, such as integrated text mining with high-throughput data analysis and integrated mining with pathway databases, are introduced. PMID- 19135550 TI - Alkoxyresorufin (methoxy-, ethoxy-, pentoxy- and benzyloxyresorufin) O-dealkylase activities by in vitro-expressed cytochrome P450 1A4 and 1A5 from common cormorant (Phalacrocorax carbo). AB - Here we report the inter-paralog comparison of cytochrome P4501A (CYP1A) catalytic function in common cormorant (Phalacrocorax carbo) using the recombinant proteins synthesized by yeast-based vector system. CYP1A4 and CYP1A5 proteins from common cormorant were heterologously expressed in yeast Saccaromyces cerevisiae. Kinetic analyses revealed that among alkoxyresorufin (methoxy-, ethoxy-, pentoxy- and benzyloxyresorufin) O-dealkylase (AROD) activities V(max) value for ethoxyresorufin O-deethylase (EROD) activity was the highest for both enzymes, reaching 0.91+/-0.034 and 1.8+/-0.043 nmol/min/nmol CYP for CYP1A4 and CYP1A5, respectively. Similar results were obtained for the catalytic efficiencies represented as the ratios of V(max) to K(m) (V(max)/K(m)). Meanwhile, distinct substrate preferences were also observed; CYP1A4 had V(max) and V(max)/K(m) values for benzyloxyresorufin O-debenzylase (BROD) activity 12- and 46-fold greater than CYP1A5, respectively, while CYP1A5 was about 13- and 4.5 fold more efficient in methoxyresorufin O-demethylase (MROD) activity than CYP1A4. The K(m) values showed no significant change among MROD, EROD, pentoxyresorufin O-depenthylase (PROD) and BROD activities for both enzymes, except for significant differences between PROD and other three activities for CYP1A4. Comparing the results in the present study with previous studies addressing chicken and rat CYP1A enzymes, it is also clear that CYP1A orthologs have different catalytic preferences for AROD activities between cormorant and rat and even between cormorant and chicken. Variations in CYP1A catalytic function between cormorant CYP1A paralogs and between CYP1A orthologs from cormorant and other species indicate that enzymatic properties should be characterized on the basis not only of a limited model species such as chicken, but also of multiple species to further understand the mechanism underlying differences in substrate selectivity and the interaction with environmental contaminants in avian species. PMID- 19135551 TI - Automatically extracting cancer disease characteristics from pathology reports into a Disease Knowledge Representation Model. AB - We introduce an extensible and modifiable knowledge representation model to represent cancer disease characteristics in a comparable and consistent fashion. We describe a system, MedTAS/P which automatically instantiates the knowledge representation model from free-text pathology reports. MedTAS/P is based on an open-source framework and its components use natural language processing principles, machine learning and rules to discover and populate elements of the model. To validate the model and measure the accuracy of MedTAS/P, we developed a gold-standard corpus of manually annotated colon cancer pathology reports. MedTAS/P achieves F1-scores of 0.97-1.0 for instantiating classes in the knowledge representation model such as histologies or anatomical sites, and F1 scores of 0.82-0.93 for primary tumors or lymph nodes, which require the extractions of relations. An F1-score of 0.65 is reported for metastatic tumors, a lower score predominantly due to a very small number of instances in the training and test sets. PMID- 19135552 TI - Endoscopic ultrasound fine-needle aspiration detection of extravascular migratory metastasis from a remotely located pancreatic cancer. AB - BACKGROUND & AIMS: Tumor cell migration along the periphery of blood vessels to remote sites has been termed extravascular migratory metastasis, which is distinct from direct gross tumor infiltration of blood vessels and from intravascular dissemination. Our objective was to report the presence of malignant perivascular cuffing of the celiac axis in a patient with an apparently early T stage and resectable pancreatic cancer. METHODS: Case report is presented. RESULTS: Endoscopic ultrasound (EUS) examination was performed with targeted fine-needle aspiration (FNA) of previously unrecognized perivascular cuffing by computed tomography, which established the presence of celiac axis malignant perivascular cuffing in the setting of a T1 pancreatic cancer. CONCLUSIONS: EUS FNA might allow identification and tissue confirmation of otherwise unrecognized extravascular migratory metastasis. This finding suggests the potential for EUS FNA to further improve pancreatic cancer staging and to enhance patient care and outcomes. PMID- 19135553 TI - MicroRNA miR-7 is preferentially expressed in endocrine cells of the developing and adult human pancreas. AB - MicroRNAs (miRNA) are small non-coding RNAs that inhibit gene expression through binding to complementary messenger RNA sequences. miRNAs have been predicted to target genes important for pancreas development, proper endocrine cell function and metabolism. We previously described that miRNA-7 (miR-7) was the most abundant and differentially expressed islet miRNA, with 200-fold higher expression in mature human islets than in acinar tissue. Here we have analyzed the temporal and spatial expression of miR-7 in human fetal pancreas from 8 to 22 weeks of gestational age (wga). Human fetal (8-22wga) and adult pancreases were processed for immunohistochemistry, in situ hybridization, and quantitative RT PCR of miRNA and mRNA. miR-7 was expressed in the human developing pancreas from around 9wga and reached its maximum expression levels between 14 and 18wga, coinciding with the exponential increase of the pancreatic endocrine hormones. Throughout development miR-7 expression was preferentially localized to endocrine cells and its expression persisted in the adult pancreas. The present study provides a detailed analysis of the spatiotemporal expression of miR-7 in developing human pancreas. The specific localization of miR-7 expression to fetal and adult endocrine cells indicates a potential role for miR-7 in endocrine cell differentiation and/or function. Future functional studies of a potential role for miR-7 function in islet cell differentiation and physiology are likely to identify novel targets for the treatment of diabetes and will lead to the development of improved protocols for generating insulin-producing cells for cell replacement therapy. PMID- 19135554 TI - Limited genetic variation in the Plasmodium falciparum heme detoxification protein (HDP). AB - Malaria parasites infecting host red blood cells degrade hemoglobin by detoxifying heme into hemozoin. This conversion of heme to hemozoin is performed by a potent protein called heme detoxification protein (HDP), making HDP an attractive target for antimalarial drug development. We studied the genetic variation in Plasmodium falciparum HDP and also investigated if HDP due to its involvement in the heme detoxification pathway is under any potential chloroquine (CQ) selection pressure. We sequenced the complete HDP gene encompassing three exons and two introns (AT and ATTT repeats in intron 1; AT repeats in intron 2) from five P. falciparum laboratory strains with known CQ sensitivity and 50 field isolates from Venezuela (n=26) and Kenya (n=24), with high levels of CQ resistance. Sequencing revealed two mutations, C41F and F91L in exon 1 and exon 2, respectively. The F41 mutation was present only in the CQ sensitive (CQS) HB3 strain. However, all the isolates harbored the 91L mutation, except for the CQS 3D7 strain. The sequencing of the intron 2 region revealed no variation in the number of AT repeats. In contrast, there was a wide variation in the AT and ATTT repeats in intron 1. Overall with respect to the intron 1 repeats, the Venezuelan isolates (Expected heterozygosity, He=0.685) showed less genetic variation as compared to the Kenyan isolates (He=0.986). Furthermore, we also genotyped the 72 76 codons of the pfcrt gene but did not observe any correlation of the pfcrt CQ resistant genotypes (SVMNT or CVIET) with variation in the HDP, thus indicating HDP not to be under any CQ selection pressure. In conclusion, HDP is a conserved target for future antimalarial development. PMID- 19135555 TI - Vitisin A suppresses LPS-induced NO production by inhibiting ERK, p38, and NF kappaB activation in RAW 264.7 cells. AB - Vitisin A, a resveratrol tetramer isolated from Vitis vinifera roots, exhibits antioxidative, anticancer, antiapoptotic, and anti-inflammatory effects. It also inhibits nitric oxide (NO) production. Here, we examined the mechanism by which vitisin A inhibits NO production in lipopolysaccharide (LPS)-induced RAW 264.7 macrophage cells. Vitisin A dose dependently inhibited LPS-induced NO production and inducible NO synthase (iNOS) expression. In contrast, the production of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) was not altered by vitisin A. To investigate the signaling pathway for NO inhibition by vitisin A, we examined nuclear factor-kappaB (NF kappaB) activation in the mitogen-activated protein kinase (MAPK) pathway, an inflammation-induced signal pathway in RAW 264.7 cells. Vitisin A inhibited LPS induced extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 phosphorylation and suppressed LPS-induced NF-kappaB activation in RAW 264.7 cells. This suggests that vitisin A decreased NO production via downregulation of ERK1/2 and p38 and the NF-kappaB signal pathway in RAW 264.7 cells. PMID- 19135556 TI - Airway nerves and dyspnea associated with inflammatory airway disease. AB - The neurobiology of dyspnea is varied and complex, but there is little doubt that vagal nerves within the airways are capable of causing or modulating some dyspneic sensations, especially those associated with inflammatory airway diseases. A major contributor to the dyspnea associated with inflammatory airway disease is explained by airway narrowing and increases in the resistance to airflow. The autonomic (parasympathetic) airway nerves directly contribute to this by regulating bronchial smooth muscle tone and mucus secretion. In addition, a component of the information reaching the brainstem via airway mechanosensing and nociceptive afferent nerves likely contributes to the overall sensations of breathing. The airway narrowing can lead to activation of low threshold mechanosensitive stretch receptors, and vagal and spinal C-fibers as well as some rapidly adapting stretch receptor in the airways that are directly activated by various aspects of the inflammatory response. Inflammatory mediators can induce long lasting changes in afferent nerve activity by modulating the expression of key genes. The net effect of the increase in afferent traffic to the brainstem modulates synaptic efficacy at the second-order neurons via various mechanisms collectively referred to as central sensitization. Many studies have shown that stimuli that activate bronchopulmonary afferent nerves can lead to dyspnea in healthy subjects. A logical extension of the basic research on inflammation and sensory nerve function is that the role of vagal sensory nerve in causing or shaping dyspneic sensations will be exaggerated in those suffering from inflammatory airway disease. PMID- 19135557 TI - Molecular basis of multidrug transport by ABC transporters. AB - Multidrug ABC transporters such as the human multidrug resistance P-glycoprotein (ABCB1) play an important role in the extrusion of drugs from the cell and their overexpression can be a cause of failure of anticancer and antimicrobial chemotherapy. These transport systems contain two nucleotide-binding domains (NBDs) where ATP is bound and hydrolyzed and two membrane domains (MDs) which mediate vectorial transport of substrates across the cell membrane. Recent crystal structures of the bacterial ABCB1 homologues Sav1866 from Staphylococcus aureus and MsbA from Salmonella typhimurium and other organisms shed light on the possible conformational states adopted by multidrug ABC transporters during transport. These structures help to interpret cellular and biochemical data gathered on these transport proteins over the past three decades. However, there are contradictory views on how the catalytic cycle of ATP binding and hydrolysis by the NBDs is linked to the change in drug binding affinity at the MDs, which underlies the capture (high affinity) of the transported drug on one side of the membrane and its release (low affinity) on the other. This review provides an overview of the current evidence for the different transport models and establishes the most recent structure-function relationships in multidrug ABC transporters. PMID- 19135558 TI - Strawberry proteome characterization and its regulation during fruit ripening and in different genotypes. AB - Strawberry is worldwide appreciated for its unique flavour and as a source of macronutrients and high levels of antioxidants which are closely related to fruit ripening. We report the investigation of the complex physiological processes of strawberry fruit ripening at proteomic level. Multiple approaches were used to investigate strawberry fruit proteome. In particular, a proteome reference map of strawberry fruit from Queen Elisa elite genotype was achieved by 2-D analyses of proteins extracted from berries at immature, turning and red stages to isolate a set of proteins commonly present in fruit during ripening. In addition, several hundreds of proteins were identified by a combination of multidimensional liquid chromatography/tandem mass spectrometry and one dimensional SDS-PAGE coupled with nano-liquid chromatography/tandem mass spectrometry. DIGE technology was also used to identify differentially accumulated proteins during ripening and to correlate fruit protein expression with quality traits of the reference variety Queen Elisa and its parental genotypes. A number of constitutive or differentially accumulated proteins were found. Generally, the pattern of protein expression as well as the putative function of identified proteins argues for a role in major fruit physiological developmental and ripening processes. The role of some of the identified proteins is discussed in relation to strawberry fruit ripening and to quality traits. Consequently, this study provides the first characterization of the strawberry fruit proteome and the time course of variation during maturation by using multiple approaches. PMID- 19135559 TI - Phosphorylated H3S10 occurs in distinct regions of the nucleolus in differentiated leaf cells. AB - Serine 10 phosphorylation of histone H3 (H3S10ph) has long been considered a mitotic marker, which is often associated with chromosome condensation both in plants and animals. Yet, in animal cells, H3S10ph was found associated with transcriptional activation of genes. Here we extend this view to plant cells showing that H3S10ph not only occurs in dividing cells during mitosis, but also in differentiated mesophyll cells. In these cells H3S10ph displayed a peculiar localization within the nucleolus where it was restricted to specific domains reminiscent of fibrillar centers. Chromatin immunoprecipitation analysis showed that H3S10ph is associated with ribosomal DNAs. Thus, in plants H3S10ph appears to be associated with two structurally differing nuclear sites engaged in gene silencing (mitotic centromeres) and in gene transcription (nucleolus). PMID- 19135560 TI - Rate of proximal humeral fractures in older Finnish women between 1970 and 2007. AB - Low-trauma fractures of older women are a major public health problem. Nevertheless, nationwide information on recent trends of proximal humeral fractures is sparse. We assessed the current trend in the number and rate (per 100,000 persons) of low-trauma fractures of the proximal humerus among 80-year old or older women in Finland, a European Union country with a well-defined white population of 5.3 million, by taking into account all women who were admitted to our hospitals for primary treatment of such fracture in 1970-2007. The number of low-trauma fractures of the proximal humerus among 80-year-old or older Finnish women rose continuously between 1970 (32 fractures) and 2007 (478 fractures), but because of a simultaneous, sharper rise in population at risk, the age-adjusted fracture rate (showing a clear rise from 88 fractures per 100,000 persons in 1970 to 304 fractures in 1995) became stabilized between 1995 and 2007 (298 fractures per 100,000 persons in 2007). In conclusion, the clear rise in the rate of low trauma fractures of the proximal humerus in Finnish elderly women from early 1970s until mid 1990s has been followed by stabilized fracture rates. Reasons for this are largely unknown, but a cohort effect toward a healthier aging population with improved functional ability and reduced risk of injurious falls cannot be ruled out. PMID- 19135561 TI - Lipid interactions, domain formation, and lateral structure of membranes. PMID- 19135562 TI - Science prevails: abortion and mental health. PMID- 19135563 TI - Two-pill regimens of misoprostol after mifepristone medical abortion through 63 days' gestational age: a randomized controlled trial of sublingual and oral misoprostol. AB - BACKGROUND: A 400 mcg dose of sublingual misoprostol has high efficacy and few side effects when used with 200 mg mifepristone for medical abortion through 63 days' gestation. STUDY DESIGN: Eligible and consenting women (n=480) were randomized to 400 mcg of misoprostol sublingually or orally, 24 h after 200 mg of mifepristone. Abortion status was assessed two weeks later. RESULTS: Complete abortion occurred in 98.7% of the sublingual group and 94.0% of the oral group (p value=.006, RR: 1.05, 95% CI=1.01--1.09). Over 90% of women in both arms expressed high satisfaction with the method. Side effects were similar in both groups, with only fever or chills reported by significantly more women in the sublingual arm. CONCLUSIONS: The sublingual route appears superior to the regimen of 400 mcg misoprostol used orally and may be a good option for mifepristone medical abortion. PMID- 19135565 TI - Analysis why nulliparous women over age 33 wish to use contraception. AB - BACKGROUND: In industrialized countries, the average age of first childbirth is increasing and more women are having no children. An understanding of the reasons for delay might provide strategies aimed at reversing this trend. STUDY DESIGN: A questionnaire survey of 234 nulliparous women aged 34 and over attending a family planning clinic in Scotland in 2006 for contraception to determine fertility intentions; awareness of fertility decline with age; and factors influencing the decision to delay childbearing. RESULTS: One hundred and sixteen women (49.6%) definitely/possibly wanted children; 118 women (50.4%) did not. Of those wanting children, 71% were concerned or very concerned about their future fertility. Most were realistic about the time it may take to conceive. Seventy-four percent of women who definitely/may want children gave reasons to do with their relationship/s as the most common reason for delay. The second most common reason was having other distractions in life. Only 34% of those wanting children, and 15% of those who did not, reported that work/training issues caused the delay. CONCLUSION: Most women were aware of the risks of delaying childbirth; however, the most common reason for delay concerned lack of the 'right' partner, something which does not lend itself to intervention. PMID- 19135564 TI - Can I get pregnant from oral sex? Sexual health misconceptions in e-mails to a reproductive health website. AB - BACKGROUND: This study identifies sexual and reproductive health misconceptions contained in e-mails sent to an emergency contraception website. STUDY DESIGN: From July 1, 2003, through June 30, 2004, 1134 English-language questions were e mailed to http://ec.princeton.edu. We performed content analysis on these e-mails and grouped misconceptions into thematic categories. RESULTS: Of the questions sent during the study period, 27% (n=303, total N=1134) evinced underlying misconceptions about sexual and reproductive health issues. Content analysis revealed five major thematic categories of misconceptions: sexual acts that can lead to pregnancy; definitions of "protected" sex; timing of pregnancy and pregnancy testing; dangers that emergency contraceptives pose to women and fetuses; and confusion between emergency contraception and abortion. CONCLUSIONS: These misconceptions have several possible sources: abstinence-only sexual education programs in the US, the proliferation of medically inaccurate websites, terminology used in public health campaigns, non-evidence-based medical protocols and confusion between emergency contraception and medication abortion in the media. PMID- 19135566 TI - Health insurance coverage and prescription contraceptive use among young women at risk for unintended pregnancy. AB - BACKGROUND: This study investigates the relationships between health insurance coverage and prescription contraceptive use among women 18-24 years of age at risk for unintended pregnancy. STUDY DESIGN: Data from the 2002 National Survey of Family Growth were analyzed. The sample comprised 1049 women 18-24 years of age at risk for unintended pregnancy. Multivariate logistic regression analysis was employed to examine the likelihood of prescription contraceptive use (1) between uninsured women versus ones with private insurance, Medicaid or another form of government insurance and (2) between those who had consistent versus inconsistent insurance coverage, after adjusting for a range of sociodemographic and sexual health factors. RESULTS: Twenty percent of the participants were uninsured and over 30% had inconsistent coverage. Only 55% were currently using prescription contraceptives. In multivariate analyses, young women with private insurance or Medicaid were more likely than the uninsured to use prescription contraceptives. No significant differences in prescription contraceptive use were found between other forms of government insurance and uninsurance or between consistent and inconsistent coverage. CONCLUSIONS: Access to comprehensive health insurance should be considered one strategy to address the high rates of unintended pregnancy among this vulnerable population. PMID- 19135567 TI - Insulin sensitivity and lipid metabolism with oral contraceptives containing chlormadinone acetate or desogestrel: a randomized trial. AB - BACKGROUND: Second-generation and third-generation oral contraceptives containing 30 mcg or more of ethinylestradiol (EE) decrease insulin sensitivity (SI). In this study, we investigated whether SI is decreased by contraceptives containing lower doses EE or by progestins with antiandrogenic properties. STUDY DESIGN: Twenty-eight young healthy women were randomly allocated to receive 20 mcg of EE and 150 mcg of desogestrel (DSG) (n=14) or 30 mcg of EE and 2 mg of chlormadinone acetate (CMA) (n=14) for 6 months. SI and glucose utilization independent of insulin (Sg) were investigated by the minimal model method. Lipid modifications were also analyzed. RESULTS: SI decreased with EE/DSG (7.09+/-1.4 vs. 4.30+/ 0.91; p=.04; n=12), but not with EE/CMA (5.79+/-0.93 vs. 6.79+/-1.1; p=.48; n=12). SI modifications observed in the two groups were significantly different ( 2.79+/-1.15 vs. 1.0+/-1.38; p=.05). Sg did not vary with either treatment. The response of C-peptide to glucose increased, but significantly so only with EE/CMA (p=.01). The C-peptide/insulin response increased with both EE/DSG (p=.05) and EE/CMA (p=.04). High-density lipoprotein (HDL) cholesterol (p=.02) and triglycerides (p=.02 and p=.01) increased in both groups, but HDL/low-density lipoprotein cholesterol (p=.02), apoprotein A1 (Apo-A1) (p=.04) and Apo A1/apoprotein B (p=.048) increased significantly only with EE/CMA. CONCLUSIONS: The present study confirms that DSG, even when associated with low EE dose, decreases SI. By contrast, EE/CMA does not deteriorate SI and induces a favorable lipid profile. PMID- 19135568 TI - Evidence that in healthy young women, a six-cycle treatment with oral contraceptive containing 30 mcg of ethinylestradiol plus 2 mg of chlormadinone acetate reduces fat mass. AB - BACKGROUND: We aimed to evaluate whether a six-cycle treatment with oral contraceptive containing 30 mcg of ethinylestradiol (EE2) plus 2 mg of chlormadinone acetate (CMA) (EE2+CMA) alters body weight (BW) and body composition of healthy young women with normal menstrual cycles. The results in treated subjects were compared to those obtained in nontreated women as control. STUDY DESIGN: Multifrequency bioelectrical impedance analysis (MF-BIA) was performed in 48 healthy young women during the follicular phase of their menstrual cycle. Of this group, 24 women were treated with EE2+CMA, and the MF BIA was repeated at the third and sixth cycle of treatment. The remaining 24 women were submitted to the same examinations after three and six cycles without any treatment. Total body water (TBW), intracellular water (ICW), extracellular water (ECW), fat mass (FM) and fat-free mass (FFM) were calculated. Waist-to-hip ratio (WHR), BW, blood pressure, and the plasma concentrations of electrolytes were also measured at each visit. RESULTS: Mean FM significantly (p<.05) decreased in the EE2+CMA group from basal levels of 14.23+/-1.03 to 13.51+/-1.09 and 12.71+/-1.02 kg at the third and sixth cycle of treatment, respectively. Stable values were seen in the control group. During observation, other parameters (BW, WHR, TBW, ECW, ICW, FFM) remained unchanged in all subjects. CONCLUSIONS: EE2+CMA reduces FM without altering TBW, ICW, ECW. These preliminary results suggest that progestational activity of CMA could balance both fluid retention and weight gain elicited by EE2. PMID- 19135569 TI - Low-dose fentanyl and midazolam in outpatient surgical abortion up to 18 weeks of gestation. AB - BACKGROUND: We investigated the safety of a conscious sedation protocol using intravenous fentanyl and midazolam by direct venous injection in women who underwent outpatient surgical abortion up to 18 weeks of gestation. STUDY DESIGN: This retrospective cohort study evaluated 1433 abortion procedures performed on women who received intravenous conscious sedation between April 1, 2001, and December 31, 2006. Women were allowed oral intake before the procedure. De identified data were abstracted from charts using a standardized extraction form. Primary outcomes evaluated were need for reversal agents, need to obtain emergency intravenous access, pulmonary aspiration, need for oxygen supplementation and hospitalization for any reason. RESULTS: Of the 1433 procedures, 410 women received sedation with continuous intravenous access, and 1023 women received sedation by direct venous injection. More than 95% of women received fentanyl 100 mcg combined with 1-2 mg of midazolam. We identified four (0.3%) instances of adverse events, none of which occurred as a result of oversedation. No women experienced oral content aspiration. CONCLUSIONS: Intravenous conscious sedation with fentanyl and midazolam is safe for outpatient surgical abortion in women without cardiovascular compromise up to 18 weeks of gestation. The risk of aspiration or oversedation requiring reversal agents is rare and does not warrant universal direct venous access or restriction of oral intake. PMID- 19135570 TI - Doxycycline serum levels at the time of dilation and evacuation with two dosing regimens. AB - BACKGROUND: Doxycycline is commonly used for antibiotic prophylaxis before dilation and evacuation (D&E) but frequently causes nausea and emesis which may affect absorption and effectiveness. Taking doxycycline the night prior to surgery may result in adequate absorption with better tolerance. STUDY DESIGN: We enrolled 40 women into a double-blind randomized comparison of doxycycline 200 mg given 4 h before D&E (Group 1) vs. the night prior to D&E with dinner (Group 2). D&E procedures were scheduled after 11 a.m. and subjects were nil per os on the morning of the procedure. Subjects completed symptom diaries from dilator placement until the D&E. Serum for doxycycline assays was obtained before the D&E. RESULTS: Mean gestational age was 19.4 weeks (range 15.8-22.0 weeks) and did not differ by group. Serum was collected at 3.2 h (range 1.9-4.8 h) and 16.3 h (range 13.8-19.1 h) after ingestion of doxycycline in Groups 1 and 2, respectively. Median serum doxycycline levels (milligrams per liter) were 2.7 and 1.8 for Groups 1 and 2, respectively (p=.04). Emesis was experienced by 50% and 15% of women in Groups 1 and 2, respectively (p=.04). Nausea ratings were worse after doxycycline in the morning compared to doxycycline with dinner and compared to placebo at either time (all p<.01). Emesis following doxycycline consumption was not associated with lower doxycycline levels (p>.2). CONCLUSION: When given with food on the night prior to D&E, doxycycline results in less emesis and nausea, but results in lower serum levels at the time of D&E. PMID- 19135571 TI - Menstrual blood loss in women using the frameless FibroPlant LNG-IUS. AB - BACKGROUND: This study was conducted to report on a menstrual blood loss (MBL) study and iron stores in women with and without heavy menstrual bleeding using the frameless FibroPlant levonorgestrel-releasing intrauterine system (LNG-IUS) for contraception. STUDY DESIGN: An open, prospective, noncomparative MBL study, using FibroPlant releasing 14 mcg of LNG/day for contraception. MBL was assessed by the quantitative alkaline hematin (QAH) technique. RESULTS: The MBL study was conducted in 40 heavy and normally menstruating Brazilian women seeking contraception. MBL was reduced from a mean baseline menstrual volume of 29.7 mL to a mean volume of 1.5 mL after 24 months, while ferritin values increased from a mean value of 31.1 ng/mL (at baseline) to a mean level of 72.5 ng/mL (after 24 months of use). Differences were highly significant (p<.0005). There were no significant differences between those who had normal menstrual bleeding and the heavy bleeders. The heavy bleeders had comparable MBL to the normal bleeders 3 months after insertion, and by 24 months post-insertion, their ferritin levels were comparable to those of the normal bleeders. Amenorrhea occurred in 80% of women out of 40 after 24 months of use. No pregnancies were recorded. CONCLUSION: The LNG-IUS is effective in reducing MBL in normally menstruating women as well as in women with heavy menstrual bleeding. The authors agree with the recommendations by the UK National Institute for Clinical Effectiveness and the new Finnish guidelines for heavy menstrual bleeding that the LNG-IUS should be positioned as first-line treatment prior to endometrial ablation or hysterectomy. PMID- 19135572 TI - Effects of progesterone, levonorgestrel and medroxyprogesterone acetate on apoptosis in human endometrial endothelial cells. AB - BACKGROUND: We evaluated apoptosis in human endometrial endothelial cells (HEECs) incubated with progesterone, levonorgestrel (LNG) and medroxyprogesterone acetate (MPA). STUDY DESIGN: HEECs were cultured to near confluence, and the progestogens were added. SETTING: Academic Department of Obstetrics and Gynecology. PATIENTS: No patients were involved. INTERVENTIONS: Progestogens at 5-, 250- and 500-ng/mL concentrations were added to incubations of HEECs for 12, 24 and 48 h. MAIN OUTCOME MEASURE: Apoptosis based on terminal deoxynucleotidyl transferase mediated deoxy-UTP nick-end labeling (TUNEL), and semiquantification of Bax and Bcl-2. RESULTS: No apoptosis was found by TUNEL, Bax and Bcl-2 after 12 h incubation with any progestogen. TUNEL increased after incubation for 24 and 48 h with progesterone 500 ng/mL; LNG 250, 500 ng/mL and all concentrations of MPA (p<.001), Bax increased and Bcl-2 decreased at all concentrations of MPA and the two highest concentrations of LNG at 48 h (p<.05). CONCLUSION: MPA results in apoptosis of HEECs. PMID- 19135573 TI - Sexual activity and contraceptive use among young female students of tertiary educational institutions in Ilorin, Nigeria. AB - BACKGROUND: A survey was conducted to evaluate the pattern of sexual behavior and contraceptive use among female students aged 15 to 24 years attending tertiary institutions in Ilorin, Nigeria. STUDY DESIGN: A valid and reliable semistructured questionnaire was self-administered to a sampled population of 600 students aged 15 to 24 years. RESULTS: Of the 600 students, 562 (93.7%) completed the questionnaire. Most (98.6%) of the respondents were unmarried, 77.6% have had sexual intercourse, 67.8% have had an unwanted pregnancy while 63.5% have had induced abortion. All the respondents were aware of contraceptives, but only 25.4% have ever used any contraceptive method. The most common sources of information about contraception among the respondents were friends/relatives (73.7%), while the fear of side effects of modern contraceptives was the most common reason (77.5%) for nonuse. CONCLUSION: The fear of side effects is the main reason for low contraceptive prevalence among young female students of tertiary institutions in Ilorin. Reproductive health services should focus more on delivery of adequate and accurate information about contraceptives to improve use among young women. PMID- 19135574 TI - Effect of sexual intercourse on the absorption of levonorgestrel after vaginal administration of 0.75 mg in Carraguard gel: a randomized, cross-over, pharmacokinetic study. AB - BACKGROUND: The Population Council studied a pre-coital contraceptive microbicide vaginal product containing levonorgestrel (LNG) as active component and Carraguard gel as a vehicle (Carra/LNG gel) for couples who engage in occasional unplanned intercourse. The objective of this study was to evaluate the effect of sexual intercourse after vaginal application of Carra/LNG gel on serum levels of LNG in women and to assess LNG absorption by the male partner. STUDY DESIGN: This was a randomized, cross-over, pharmacokinetic study including an abstinence arm and an arm in which couples engaged in sexual intercourse between 2 and 4 h after gel application. In each study arm, each woman received a single application of Carra/LNG gel (0.75 mg in 4 mL gel) followed by serial blood samples taken at 0, 1, 2, 4, 8, 24 and 48 h after gel application for LNG measurements. In the intercourse arm, LNG was measured in blood samples taken from the male partner before intercourse and at 4, 8 and 24 h after gel application in the female partner. RESULTS: Time concentration curves for serum LNG levels showed a mean C(max) of 7.8+/-5.5 and 8.3+/-5.7 nmol/L, a mean T(max) of 6.2+/-5.9 and 7.5+/ 5.7, and comparable area under the curve for the intercourse and abstinence arm, respectively. Pharmacokinetic parameters presented large variability between subjects, but excellent reproducibility within each subject. LNG was undetectable in 10 out of 12 male partners. CONCLUSION: Sexual intercourse does not appear to interfere with vaginal absorption of LNG after application of a Carra/LNG gel. A vaginal pre-coital contraceptive gel is feasible. PMID- 19135575 TI - Ectopic pregnancy following levonorgestrel emergency contraception: a case report. AB - Use of levonorgestrel as emergency contraception is a safe and effective measure to prevent unwanted pregnancy. However, ectopic gestation in case of failure is a known risk. Access to levonorgestrel without a prescription in many countries has made it impossible to estimate the exact incidence of this potential adverse event. Thus, spontaneous reporting of cases serves to alert physicians to this possibility. We present a case of ectopic pregnancy following use of levonorgestrel emergency contraception. To our knowledge, this is the first case report from India following introduction of levonorgestrel emergency contraception in 2001. PMID- 19135576 TI - Serious adverse events associated with the use of misoprostol alone for cervical preparation prior to early second trimester abortions (12-16 weeks). PMID- 19135577 TI - Pediatric Clinics of North America. Common respiratory symptoms and illnesses: a graded evidence based approach. Preface. PMID- 19135578 TI - Respiratory noises: how useful are they clinically? AB - Although clinicians place considerable weight on the identification of the various forms of noisy breathing, there are serious questions regarding both the accuracy (validity) and the reliability (repeatability) of these noises. To avoid diagnostic errors, clinicians need to consider the whole constellation of symptoms and signs, and not focus on the specific "type" of noise. Given the high error rate with "parent-reported wheeze" there is a need to reexamine the extensive literature on the epidemiology of wheeze in infants and young children, because parent-reported wheeze is unconfirmed by a clinician. It is obvious we need more high-quality research evidence to derive better evidence on the clinical utility of these noises, and their natural history. PMID- 19135579 TI - Cough. AB - The management of cough in children should be etiologically based. This requires that all children with cough should be carefully evaluated and managed differently than adults because the etiologic factors and treatment in children are significantly different than that in adults. In all children with cough, exacerbation factors should be sought and intervention options for cessation advised or initiated. Parental expectations and specific concerns should also be sought and addressed. PMID- 19135580 TI - Perceptions and pathophysiology of dyspnea and exercise intolerance. AB - Dyspnea is a complex psychophysiologic sensation that requires intact afferent and efferent pathways for the full perception of the neuromechanical dissociation between the respiratory effort attempted and the work actually accomplished. The sensation is triggered or accentuated by a variety of receptors located in the chest wall, respiratory muscles, lung parenchyma, carotid body, and brain stem. The sensation of dyspnea is stronger in patients with higher scores for anxiety and has been reported in patients with anxiety disorders with no cardiopulmonary disease. These observations demonstrate the importance of cerebral cognition in this complex symptom. Ten cases are presented that illustrate different clinical manifestations of dyspnea. PMID- 19135581 TI - Chest pain and chest wall deformity. AB - Chest pain and chest wall deformities are common in children. Although most children with chest pain have a benign diagnosis, some have a serious etiology for pain, so the complaint must be addressed carefully. Unfortunately, there are few prospective studies to evaluate this complaint in children. Serious causes for chest pain are rare, making it difficult to develop clear guidelines for evaluation and management. The child who appears well, has a normal physical examination, and lacks worrisome history deserves reassurance and careful follow up rather than extensive studies. Multicenter studies are needed to better define this important symptom. PMID- 19135582 TI - Recurrent respiratory infections. AB - The child who has recurrent infections poses one of the most difficult diagnostic challenges in pediatrics. The clinician faces a two-fold challenge in determining first whether the child is normal or has a serious disease, and then, in the latter case, how to confirm or exclude the diagnosis with the minimum number of the least invasive tests. It is hoped that, in the absence of good-quality evidence for most clinical scenarios, the experience-based approach described in this article may prove a useful guide to the clinician. PMID- 19135583 TI - Upper respiratory tract infections (including otitis media). AB - Upper respiratory tract infections (including otitis media) are the most common illnesses affecting children. Most illnesses are mild and resolve completely without specific treatment, but the frequency of infection and association with fever and constitutional symptoms creates significant distress for the child and the family. By understanding the evidence available from high-quality studies, the clinician can advise the families on appropriate action. The goal of this article is to support clinicians in answering the following questions: (1) What happened to children with these conditions when no additional treatment was provided? (2) Which interventions have been assessed in well-designed studies? (3) Which interventions have been shown to improve outcomes? (4) How large is the overall benefit? PMID- 19135584 TI - Acute bronchiolitis and croup. AB - Croup and acute bronchiolitis are common forms of virally induced respiratory disease in infancy and early childhood. There is good evidence that corticosteroids can ameliorate disease severity and alter the natural history of symptoms in patients who have croup and that temporary symptomatic benefit can be obtained from the use of nebulized adrenaline. The principle weakness when reviewing therapeutic interventions for acute bronchiolitis is the lack of a clear diagnostic test or definition. Current evidence suggests that oxygen is the only useful pharmacologic agent for correcting hypoxia. PMID- 19135585 TI - Pneumonia and other respiratory infections. AB - Pneumonia is a leading killer of children in developing countries and results in significant morbidity worldwide. This article reviews the management of pneumonia and its complications from the perspective of both developed and resource-poor settings. In addition, evidence-based management of other respiratory infections, including tuberculosis, is discussed. Finally, the management of common complications of pneumonia is reviewed. PMID- 19135586 TI - Bronchiectasis in children. AB - Bronchiectasis is a chronic disease of the conducting airways that produces persistent productive cough, recurrent respiratory infectious exacerbations, and obstructive lung disease in children and adults. This article focuses on the grading and recommendations for chronic therapies of bronchiectasis caused by cystic fibrosis (CF)- and non-CF-related conditions. The scope of this article is to focus on outpatient treatment and not include as-needed treatment for mild or severe pulmonary exacerbations associated with bronchiectasis. PMID- 19135587 TI - Aspiration lung disease. AB - The term aspiration lung disease describes several clinical syndromes, with massive aspiration and chronic lung aspiration being at two extremes of the clinical spectrum. Over the years, significant advances have been made in understanding the mechanisms underlying dysphagia, gastroesophageal function, and airway protective reflexes and new diagnostic techniques have been introduced. Despite this, characterizing the presence or absence of aspiration, and under what circumstances a child might be aspirating what, is extremely challenging. Many children are still not adequately diagnosed or treated for aspiration until permanent lung damage has occurred. A multidisciplinary approach is mandatory for a correct diagnosis in addition to timely and appropriate care. PMID- 19135588 TI - Asthma in childhood. AB - Management decisions for pediatric asthma (in patients younger than 12 years of age) based on extrapolation from available evidence in adolescents and adults (age 12 years and older) is common but rarely appropriate. This article addresses the disparity in response between the two age groups, presents the available pediatric evidence, and highlights the important areas in which further research is required. Evidence-based recommendations for acute and interval management of pediatric asthma are provided. PMID- 19135589 TI - Congenital airway lesions and lung disease. AB - Structural upper and lower airway disorders and parenchymal disorders are uncommon in pediatric practice, but many pediatricians will encounter them and be responsible for the ongoing care of these patients. Pediatricians need to be cognizant of these diagnoses because, even though management of these disorders generally lacks an evidence base, existing principles of good care surrounding accurate diagnosis, classifications of severity, judicious use of investigations, medication, and surgical approaches are essential to good outcomes. PMID- 19135590 TI - Obstructive sleep breathing disorders. AB - Obstructive sleep apnea (OSA) is increasingly recognized in children. There is accumulating robust evidence to support early diagnosis and treatment of this condition. The purpose of this review is to provide an update on the epidemiology, clinical features, complications, and treatment of childhood OSA. The authors have also proposed an easy-to-follow flowchart on the management of children with snoring or sleep disturbance for clinical use by busy pediatricians. PMID- 19135591 TI - Chronic respiratory failure and neuromuscular disease. AB - The outlook for children with respiratory complications of neuromuscular disease has improved significantly in the past 15 years. This has been the result of many advances in clinical care, including improved monitoring of lung function and hypoventilation during sleep; coordinated respiratory care by experienced physicians with access to specialized respiratory services, especially physiotherapy; and, most importantly, the widespread introduction of noninvasive ventilation. PMID- 19135592 TI - Domiciliary oxygen for children. AB - Domiciliary oxygen is used increasingly in pediatric practice, and the largest patient group to receive it is ex-premature babies with chronic neonatal lung disease. Because of a scarcity of good evidence to inform clinicians, there is a lack of consensus over many issues, even those as fundamental as the optimum target oxygen saturation. Nevertheless, many children benefit from receiving supplemental oxygen at home, particularly because it helps to keep them out of the hospital. PMID- 19135593 TI - Abstracts of the 40th Annual Meeting of the Society of Gynecologic Oncologists. San Antonio, Texas, USA. February 5-8, 2009. PMID- 19135594 TI - Carry that Weight. PMID- 19135595 TI - Violent conflict: protecting the health of civilians. PMID- 19135596 TI - How good is your doctor? Click here to find out. PMID- 19135597 TI - Change4Life brought to you by PepsiCo (and others). PMID- 19135598 TI - Beta blockers for asthma: a double-edged sword. PMID- 19135599 TI - International collaborative research on Charcot's disease. PMID- 19135601 TI - Marian Jacobs: pioneer of child health in South Africa. PMID- 19135602 TI - Paracetamol as a risk factor for allergic disorders. PMID- 19135603 TI - Paracetamol as a risk factor for allergic disorders. PMID- 19135604 TI - Paracetamol as a risk factor for allergic disorders. PMID- 19135606 TI - Paracetamol as a risk factor for allergic disorders. PMID- 19135608 TI - Exenatide for type 2 diabetes. PMID- 19135609 TI - Malaria in sub-Saharan Africa. PMID- 19135610 TI - Effect of the financial crisis and rescue plan on ordinary Americans. PMID- 19135611 TI - The global financial downturn-imperatives for the health sector. PMID- 19135612 TI - Visualising rhabdomyolysis. PMID- 19135613 TI - Mechanisms of thrombogenesis in atrial fibrillation: Virchow's triad revisited. AB - Atrial fibrillation is the most common sustained cardiac arrhythmia, which is associated with a high risk of stroke and thromboembolism. Increasing evidence suggests that the thrombogenic tendency in atrial fibrillation is related to several underlying pathophysiological mechanisms. Abnormal changes in flow are evident by stasis in the left atrium, and seen as spontaneous echocontrast. Abnormal changes in vessel walls-essentially, anatomical and structural defects include progressive atrial dilatation, endocardial denudation, and oedematous or fibroelastic infiltration of the extracellular matrix. Additionally, abnormal changes in blood constituents are well described, and include haemostatic and platelet activation, as well as inflammation and growth factor changes. These changes result in the fulfilment of Virchow's triad for thrombogenesis, and accord with a prothrombotic or hypercoagulable state in this arrhythmia. In this Review, we present an overview of the established and purported mechanisms for thrombogenesis in atrial fibrillation. PMID- 19135614 TI - The insulin ball. PMID- 19135615 TI - Aldosterone and Cardiovascular Disease. Foreword. PMID- 19135616 TI - Aldosterone and cardiovascular disease. AB - Aldosterone is an adrenal hormone that regulates sodium, fluid, and potassium balance. Jerome Conn first described the syndrome of autonomous and excessive aldosterone secretion or "primary aldosteronism." Contrary to the historical belief, recent studies indicate that primary aldosteronism is a common cause of hypertension with a prevalence of 5-10% among general hypertensive patients. Various animal models have demonstrated that aldosterone in association with a high salt diet results in target-organ inflammation and fibrosis. Similarly, cross-sectional and observational human studies have demonstrated the association of aldosterone with development and severity of hypertension, congestive heart failure, coronary artery disease, chronic kidney disease, and metabolic syndrome. Several interventional studies have also demonstrated the beneficial effects of mineralocorticoid receptor antagonists in these disease processes, particularly hypertension, heart failure, and post myocardial infarction, further supporting the role of aldosterone in their pathogenesis. We review the role of aldosterone in these various cardiovascular disease processes along with potential mechanisms and treatment. PMID- 19135617 TI - First-drug treatment failures in children newly diagnosed with epilepsy. AB - In adults newly diagnosed with epilepsy, treatment with the first prescribed antiepileptic drug fails for approximately one half. In two studies that addressed this question in children, the failure rates were 20% and 40%. The present study used a detailed chart review of children newly diagnosed with epilepsy over a 4-year span in a major childhood epilepsy referral clinic to assess (1) the percentage of children for whom first-line antiepileptic drug treatment failed and (2) the reasons for the treatment failure. Charts were reviewed for 95 children who were diagnosed with epilepsy, started on their first antiepileptic drug, and then monitored for approximately 5 years. Of these 95 children, 48 were classified as having idiopathic epilepsy (50.5 %), 30 as having cryptogenic epilepsy (31.6%), and 17 as having symptomatic epilepsy (17.9%). The two main antiepileptic drugs used were valproic acid (43.2% of patients) and carbamazepine (38.9% of patients). Treatment with the first antiepileptic drug failed in 30/95 children (31.6%). Treatment failure was due to adverse effects in 12/30 children (40.0%), due to lack of efficacy in 11/30 (37.9%), and due to both adverse effects and lack of efficacy in 7/30 (24.1%). Also examined was the effect on treatment failure of patient age at diagnosis, antiepileptic drug choice, maximum drug dose, etiology of epilepsy, and particular epilepsy syndromes on treatment failures; there was no statistically significant effect of any of these variables on first-line treatment outcome. In this population, approximately one third of children newly diagnosed with epilepsy experienced treatment failure with the first antiepileptic drug used. Lack of efficacy and unacceptable adverse effects contributed equally to these treatment failures. PMID- 19135618 TI - Implementation of a febrile seizure guideline in two pediatric emergency departments. AB - Despite the typically benign nature of febrile seizures, a large number of children with simple febrile seizures are overinvestigated and overtreated, according to the personal clinical experience of the treating doctors. The study objective was to analyze the effect of implementing an evidence-based medicine guideline on the management of febrile seizures in two European pediatric emergency departments. After introduction of a selected guideline, we analyzed the change in hospitalization rate and in the rate of execution of blood exams in children presenting with febrile seizures. Included in the study were 483 children. Clinical characteristics of seizures were similar both before and after implementation. Clinical management was modified after guideline introduction, because the hospitalization rate significantly decreased (respectively, from 57.3% to 20.5%, and from 16.9% to 3.2%), without any concomitant increase in readmission rate. Readmission cases were never due to severe bacterial infections. The proportion of patients who received blood examinations decreased significantly. We conclude that in both of the Emergency Departments studied, introduction of a guideline on febrile seizure positively modified clinical management. The availability of a guideline contributed to accelerating the process of improving welfare and positively influenced the quality of care. PMID- 19135619 TI - Combined central diabetes insipidus and cerebral salt wasting syndrome in children. AB - Central diabetes insipidus, a common consequence of acute central nervous system injury, causes hypernatremia; cerebral salt wasting syndrome can cause hyponatremia. The two conditions occurring simultaneous are rarely described in pediatric patients. Pediatric cases of combined diabetes insipidus and cerebral salt wasting after acute central nervous system injury between January 2000 and December 2007 were retrospectively reviewed, and clinical characteristics were systemically assessed. Sixteen patients, aged 3 months to 18 years, met study criteria: 11 girls and 5 boys. The most common etiologies were severe central nervous system infection (n = 7, 44%) and hypoxic-ischemic event (n = 4, 25%). In 15 patients, diabetes insipidus was diagnosed during the first 3 days after acute central nervous system injury. Onset of cerebral salt wasting syndrome occurred 2 8 days after the onset of diabetes insipidus. In terms of outcome, 13 patients died (81%) and 3 survived under vegetative status (19%). Central diabetes insipidus and cerebral salt wasting syndrome may occur after acute central nervous system injury. A combination of both may impede accurate diagnosis. Proper differential diagnoses are critical, because the treatment strategy for each entity is different. PMID- 19135620 TI - Leigh syndrome: clinical and neuroimaging follow-up. AB - Leigh syndrome, caused by dysfunction in mitochondrial energy metabolism, is an inherited, heterogeneous, and progressive neurodegenerative disorder of infancy and childhood. From 1983 to August 2006, 14 cases diagnosed with Leigh syndrome were studied in terms of characteristic neuroimaging findings and abnormal mitochondrial configurations under electron microscopy, as well as molecular analysis. Of the 14 cases, 11 presented clinical features before age 1 (79%). All 14 presented with variable symptoms of central nervous system involvement. The three most common symptoms were developmental delay (12/14; 86%), seizures (11/14; 79%), and altered consciousness (8/14; 57%). Extra-central nervous system manifestations were observed in 10 of the 14 cases, the most common symptoms being failure to thrive (5/14; 36%), pericardial effusion and dilated cardiomyopathy (3/14; 21%), and liver function impairment (3/14; 21%). In all 14 cases, neuroimaging revealed abnormal findings over the basal ganglion, brainstem, or both. The putamen was the most common lesion site in the basal ganglia (11/12; 92%). Cranial magnetic resonance imaging was used for follow-up in 6 cases because of changes in clinical features; in all 6 cases the imaging revealed evolution in the brain. Cranial magnetic resonance spectroscopy was performed in 3 cases and in 2 of them revealed lactate peaks during deterioration of the disease course. The prognosis for Leigh syndrome was poor during long-term follow-up. Seven cases were early fatalities, before 1 year and 6 months of age. Follow-up cranial magnetic resonance imaging together with magnetic resonance spectroscopy in cases with clinical evolution is helpful for monitoring this disease. PMID- 19135621 TI - Effect of carbamazepine therapy on serum lipids in children with partial epilepsy. AB - Alteration in serum lipids leads to atherosclerosis. Antiepileptic drugs alter serum lipids. The effect of carbamazepine therapy on serum lipids of children with partial epilepsy and correlation of changes with carbamazepine level was prospectively studied. The study population of 29 children (16 male, 13 female; age range, 3-12 years) diagnosed with partial epilepsy were enrolled within 14 days of starting carbamazepine. Serum lipids were measured at enrollment and at 3 months. Carbamazepine levels were determined at 3 months of therapy. Mean total cholesterol was 130.2 +/- 27.3 mg/dL at enrollment, and increased significantly to 144.8 +/- 32.9 mg/dL at 3 months (P = 0.018). There was a significant change in mean low and very low density lipoproteins, and in triglycerides (P < 0.05). Both the ratio of total cholesterol to high density lipoprotein and the ratio of low density lipoprotein to high density lipoprotein increased significantly. There was no significant change in levels of high density lipoprotein. At 3 months, mean carbamazepine level was 6.5 +/- 1.8 microg/dL. Changes in serum lipids did not correlate with serum carbamazepine level (P > 0.05). Carbamazepine therapy increases levels of lipids that increase the risk of atherosclerosis. PMID- 19135622 TI - Pediatric cerebral angiography: analysis of utilization and findings. AB - Pediatric neuroendovascular procedures are being performed with increasing frequency, for various indications. Reported here is the experience of interventionally trained neurologists performing pediatric cerebral diagnostic angiography between August 1, 2005, and April 30, 2008, at a single tertiary institution. Data regarding patient demographics, diagnostic indication and angiographic diagnosis, procedural complications, and procedural specifications were recorded to assess practice patterns and to track procedural morbidity. In all, 42 patients had 46 procedures during the study period. Mean age was 9.97 years (standard deviation S.D. = 5.39; range, 0.3-18 years); 22/42 were male (52%). Known or suspected vascular malformation was the diagnostic indication for 20 patients; of these, 12 had an arteriovenous malformation, 5 had venous abnormalities, and 3 exhibited no angiographic vascular malformations. In 13 total procedures there was no angiographic pathology. General anesthesia was used in 29/46 procedures (63%). A total of 190 cerebral arteries were individually selected, with a mean number of vessels catheterized of 4.1 (S.D. = 1.7) per procedure. No procedural thromboembolic complications, iatrogenic arterial dissection, or neurologic or vascular access site complications occurred. In conclusion, pediatric cerebral angiography seems to be generally safe, although there should be a strong diagnostic indication, given the inherent procedural risk. PMID- 19135624 TI - Serum anti-nuclear antibodies as a marker of autoimmunity in Egyptian autistic children. AB - Autism may involve an autoimmune pathogenesis in a subgroup of patients. The frequency of anti-nuclear antibodies in 80 autistic children and their relationship to a family history of autoimmunity were studied, compared with 80 healthy, matched children. Children with autism had a significantly higher percent seropositivity of anti-nuclear antibodies (20%) than healthy children (2.5%; P < 0.01). Fifty percent of anti-nuclear antibody-seropositive autistic children had an anti-nuclear antibody titer of > or =1:640 (very high positive); 25%, > or =1:160 (high positive); and the remaining 25%, 1:80. All anti-nuclear antibody-seropositive healthy children had anti-nuclear antibody titers of 1:80. A family history of autoimmunity was significantly higher in autistic children (47.5%) than healthy controls (8.8%; P < 0.001). Anti-nuclear antibody seropositivity was significantly higher in autistic children with a family history of autoimmunity than those without such history (36.8% and 5%, respectively; P < 0.001). Anti-nuclear antibody seropositivity had significant positive associations with disease severity, mental retardation and electroencephalogram abnormalities. Autoimmunity may play a role in a subgroup of children with autism. Further studies are warranted to assess anti-nuclear antibody seropositivity, other markers of autoimmunity (e.g., brain-specific autoantibodies), and the role of immunotherapy in children with autism. PMID- 19135623 TI - Thrombophilic risk factors in epileptic children treated with valproic Acid. AB - There are few reports on valproic acid related to thrombophilia. Thrombophilic risk factors were investigated in 21 children (age range, 1-13 years) diagnosed with epilepsy and newly treated with valproic acid monotherapy. None of the children had been previously treated with any anticonvulsant agent. Before starting valproic acid therapy, homocysteine, lipoprotein(a), factor VIII, factor IX, protein C, protein S, antithrombin III levels, and activated protein C resistance levels were evaluated in all patients, with repeat evaluation after 9 months or 1 year of the therapy. Thrombosis gene mutations (factor V Leiden and prothrombin G20210A) were also evaluated in all patients before therapy. There was statistically significant elevation in lipoprotein(a) levels and reduction in fibrinogen levels after treatment. Reduction in protein C levels and elevation in homocysteine levels were also observed, but without statistical significance. Before therapy, no thrombotic event had occurred, despite thrombotic risk factors in some patients. Valproic acid can increase lipoprotein(a) and decrease fibrinogen, which may increase the risk of stroke or other thrombotic events. No clinical adverse effects resulted from changes in the levels or activity of thrombophilic factors associated with valproic acid treatment. Thus, routine investigation of factors implicated in thrombosis prior to initiation of valproic acid is not warranted for all patients. Nonetheless, caution is advised when initiating valproic acid treatment in children who have had prior stroke or thrombotic events. PMID- 19135625 TI - Febrile seizures: interleukin 1beta and interleukin-1 receptor antagonist polymorphisms. AB - In order to investigate the association between IL-1beta -511 C-->T and IL-1 receptor antagonist intron 2 variable tandem repeat polymorphisms, and febrile seizures in children, 90 children (mean age, 19.7 +/- 11.2 months) diagnosed with febrile seizure and 106 healthy controls (mean age, 14.2 +/- 3.6 months) with no seizure or neurologic events were included in the study. The polymorphisms were analyzed using restriction fragment length polymorphism and agarose gel electrophoresis methods. In the patient group, the frequencies of IL-1beta genotypes CC, CT, and TT were 24.4%, 52.2%, and 23.3%, respectively, compared with 38.7%, 50.95%, and 10.4%, respectively, in the control group. The TT genotype was significantly more common in the patient group than in the control group (P = 0.044), and the T allele frequency was significantly higher in the patient group (0.50 vs 0.36, P = 0.040). Among the three genotypes (RN1/1, RN1/2, and RN2/2) of the IL receptor antagonist gene variable tandem repeat polymorphisms, the frequency of both the RN2/2 genotype and the RN2 allele were significantly higher in the patient group (P = 0.007). Also RN2 allele frequency was found higher in patient group than controls (0.29 vs 0.15, P = 0.020). IL 1beta -511 and IL-1 receptor antagonist intron 2 variable tandem repeat polymorphisms may be involved in susceptibility to febrile convulsions in children. PMID- 19135626 TI - Neurofibromatosis 1 associated with spinal muscular atrophy. AB - Neurofibromatosis type 1, or von Recklinghausen disease, is a progressive, autosomal dominant, monogenic disease. Spinal muscular atrophy is a progressive, autosomal recessive, monogenic disease. Specific anti-polysaccharide antibody deficiency is an immune disorder suspected in any child older than 2 years who suffers from recurrent respiratory tract infections or in patients with unusually severe complications from infections under appropriate treatment. Reported here is the coinheritance of two monogenic syndromes in the same patient, a novel association with specific anti-polysaccharide antibody deficiency. PMID- 19135627 TI - Status epilepticus in a pediatric patient with amantadine overdose. AB - A 2-year-old boy who ingested 0.8-1.5 g of amantadine developed status epilepticus. One hour later, the child presented with agitation, diaphoresis, and vomiting. He was admitted to the pediatric emergency department 2 hours later. Generalized seizures evolved to status epilepticus, with alternating generalized tonic-clonic and partial seizures, over a period of 7 hours. Other initial clinical signs were sinusal tachycardia and reactive bilateral mydriasis. All symptoms resolved within 20 hours, with a good recovery; the child was released from the hospital on day 3. PMID- 19135628 TI - Juvenile xanthogranuloma of the cauda equina. AB - Juvenile xanthogranuloma is a form of histiocytic proliferative disorder that usually affects the skin and tends to occur during infancy. On rare occasions, it has been reported at extracutaneous sites and in other age groups. Isolated juvenile xanthogranuloma of the nervous system is extremely rare, especially in the cauda equina. A case of juvenile xanthogranuloma of the cauda equina in a 14 year-old boy is reported, and the literature is reviewed. PMID- 19135629 TI - The effect of constipation on valproic acid dosage in a 17-year-old. AB - Although valproic acid absorption measurements have been reported for animal models, there have been no reports that correlate this to clinical experience with a patient. Reported here is the case of a patient who required twice the dose of valproic acid after colostomy for chronic constipation. The increased dosage required may have been due to rapid intestinal transit time or to bypass of the left colon. This case highlights the importance of gastrointestinal function in patients on valproic acid therapy. PMID- 19135630 TI - Mycoplasma pneumoniae--associated transverse myelitis and rhabdomyolysis. AB - Mycoplasma pneumoniae is a common cause of respiratory tract infection. Extrapulmonary manifestations of M. pneumoniae infection are also common. The present case is that of a previously healthy 4-year-old boy who displayed a novel simultaneous onset of both acute rhabdomyolysis and transverse myelitis associated with an infection of M. pneumoniae. He had no preceding symptoms or signs of respiratory tract infection. Intravenous immunoglobulin (1 g/kg per day) for 2 days was prescribed initially for the deterioration of neurologic condition. His rhabdomyolysis resolved without complication, but neurologic sequelae remained during 2 years of follow-up. Evaluation for M. pneumoniae infection is recommended in patients with idiopathic rhabdomyolysis and transverse myelitis, even if in the absence of antecedent respiratory symptoms. PMID- 19135631 TI - Rotavirus-associated encephalopathy with a reversible splenial lesion. AB - The case is reported of a 2-year-old boy with a rotavirus-induced mild encephalopathy that presented as transient intensified signal on the splenium of the corpus callosum. The boy also experienced persistent diarrhea, vomiting, fever, and sudden disturbance of consciousness. Although cerebrospinal fluid analysis did not manifest pleocytosis, electroencephalography demonstrated global diffuse slow waves and cranial magnetic resonance imaging demonstrated intensified signal on the splenium of the corpus callosum. Methylprednisolone was infused for 3 days. The disturbance of consciousness disappeared within 24 hours without any other complications, and the splenial signal and electroencephalogram returned to normal within 6 days. PMID- 19135632 TI - A 30-year follow-up of a neuronal ceroid lipofuscinosis patient with mutations in CLN3 and protracted disease course. AB - Reported here is the 30-year follow-up of a patient, diagnosed with juvenile neuronal ceroid lipofuscinosis, who was compound heterozygous for the common 1-kb deletion and the missense mutation p.Glu295Lys in the CLN3 gene. Visual failure was noticed at 6 years of age, but thereafter disease progression was atypical. Polyneuropathy and cerebellar signs were observed after age 20, and epilepsy and slight mental decline after age 35. From then on, there was rapid deterioration, and the patient died at age 39. This case highlights the importance of exact genotyping for disease course prediction and management. PMID- 19135633 TI - Nonketotic hyperglycinemia and acquired hydrocephalus. AB - Nonketotic hyperglycinemia is an autosomal recessive disorder of glycine metabolism. Patients generally present in the neonatal period with lethargy, feeding difficulty, hypotonia, apnea, poorly controlled convulsions, and coma. Myoclonic seizures and burst suppression pattern on electroencephalography are major findings of disease, but development of hydrocephalus is not an expected finding. The present case is that of an infant with acquired hydrocephalus, psychomotor retardation, and myoclonic seizures in whom the final diagnosis was nonketotic hyperglycinemia. PMID- 19135634 TI - Biosensors 2008: the 10th world congress on biosensors. PMID- 19135635 TI - [Erectile dysfunction and renal chronic insufficiency: etiology and management]. AB - Erectile dysfunction (ED) has a higher incidence in patients treated for chronic renal insufficiency or in patients who underwent kidney transplanation as it concerns more than 50% of them. Its severity is directly linked with the seriousness of the renal disease. ED is responsible of a deterioration of the quality of life. ED's physiopathology is complex and multifactorial, involving a combination of classical risk factors (endothelial dysfunction), specific factors (e.g., chronic hyperuremia and co-morbidities) and psychological factors. Management of ED must take into account both sides of the disease in order to propose appropriate treatment; i.e, psychological concerns and organic matters. Although literature remains poor in this area, phosphodiesterase-5 inhibitors are increasingly used for these patients as they are safe and efficient most of the time. Pharmacokinetics of phosphodiesterase-5 inhibitors can be disturbed by the simultaneous use of immunosuppressor. As a second line, intra-cavernous injections remain a gold-standard treatment. In case of failure, penile prosthesis can even be considered in case of renal chronic insufficiency and in transplanted patients. PMID- 19135636 TI - [Atypical cysts and cystic tumours of the kidney: histological, radiological and surgical considerations. Conclusions of the AFU 2007 forum]. AB - Malignant tumours may have a cystic appearance. They are dominated by multilocular cystic renal cell carcinoma, usually low-grade, which rarely metastasize. The Bosniak classification distinguishes non suspicious lesions (type I and II) from suspicious lesions (type III and IV) requiring resection and lesions requiring follow-up (type IIF). The main feature suggestive of malignancy is the enhancement of the septa and the walls of the cyst. Renal cysts classified as IIF require surveillance by contrast-enhanced imaging (CT, MRI or ultrasound). The treatment of cystic tumours is based on surgery. Partial nephrectomy is recommended in this type of tumour regardless of the size. Laparoscopy is a validated technique in experienced hands. Aspiration is not very effective for the treatment of benign cysts, but may be useful for diagnosis. Surgical resection of the roof of the cyst is the most effective technique. PMID- 19135637 TI - [Renal stab wounds: about 20 cases]. AB - OBJECTIVE: To assess the indication for operative and nonoperative management of renal stab wounds in patients hospitalised over the last six years. PATIENTS AND METHODS: Retrospectively studied of 20 cases over the last six years with penetrating kidney trauma, managed in two urologic departments of Marrakech and Casablanca. We studied the signs on admission (haematuria, blood pressure, haemoglobin and serum creatinine), associated lesions, treatment and complications. The imaging procedures comprised abdomen ultrasonography and abdomen computed tomography scan (CT). RESULTS: The mean age was 25-year-old (range: 14-50 years), macroscopic hematuria was present in 95%. The weapon introduction site was the left lumbar fossa in 15 cases, the right lumbar fossa in four cases and the left flank in one case. Six patients (30%) presented with haemodynamic instability. Anaemia (<10g/dl) was found in 14 cases (70%) and blood transfusion was necessary in eight cases (40%). According to the American Association of Trauma Surgery (AATS) classification of kidney trauma, our patients were classified as follows: Grade I (two cases), Grade II (four cases), Grade III (six cases), Grade IV (five cases) and Grade V (three cases). The conservative management was adopted for 15 patients (75%), and two patients had a double pigtel ureteric stent for an important leakage of the contrast product. A CT scan was systematically performed ten days after the trauma and the kidney traumas lesions were often stabilized. Nephrectomy was performed for 5 patients (25%) grade IV (two cases) and grade V (three cases). The patients were discharged after a mean period of 12 days (six to 33 days). CONCLUSION: The development of interventional radiology, endourological drainage techniques and medical intensive care helps to limit the complications and to manage conservatively with a correct resuscitation for selected patient. A grade V is a surgical indication. PMID- 19135638 TI - [Endoscopic removal of renal stones through laparoscopic access of the ureter and the pelvis]. AB - PURPOSE: We studied the efficiency and the morbidity of endoscopic removal of kidney stones during laparoscopic removal of ureteral stone. MATERIAL AND METHODS: Six patients presenting with an ureteral stone resisting to ESWL and four patients presenting with an UPJ obstruction were studied. Every patient had one to five associated kidney stones. The patients were operated with transperitoneal laparoscopic ureterolithotomy or pyeloplasty. An endoscopy of the upper urinary tract was realized during the same operating time through laparoscopic access. A flexible ureteroscope (five cases) or semi-flexible ureteroscope (one case) were used after ureterolithotomy. A fibroscope (four cases) was used before pyeloplasty. The endoscope was introduced through the port of the iliac fossa and the stones were extracted with a basket grasper. The patients were followed by abdominal plain film 12 weeks after surgery, by IPV six weeks after surgery and then annually with abdominal plain film and ultrasound. RESULTS: No operative complication arose. Nine of 10 patients had a complete extraction of the kidney stones (one patient had an extraction only of four out of five kidney stones). No damage of any endoscope was observed. At a minimum follow-up of 18 months, no fistula nor stenosis of the ureter was diagnosed. CONCLUSION: Endoscopic removal of kidney stones through laparoscopic access of the upper urinary tract is effective, sure and reproducible. Such procedure requires experience in laparoscopy and endoscopy of the upper urinary tract. The evaluation of this procedure must be pursued. PMID- 19135639 TI - [Retrograde endopyelotomy using Holmium-Yag laser for uretero-pelvic junction obstruction]. AB - OBJECTIVE: To evaluate our results of retrograde laser endopyelotomy for uretero pelvic junction obstruction. MATERIAL AND METHODS: Retrospective study of 27 consecutive retrograde laser endopyelotomies performed on 24 patients over a six years period (June 1999 to July 2005). Sixteen stenoses were primary. The level of obstruction was severe in 13 patients and moderate in 14 patients. A polar pedicle was diagnosed by pre-operative CT-angiography in seven cases. Balloon dilatation was performed in 17 procedures. A double J ureteral stent remained in place for six weeks mean. We evaluated results by a clinical examination and an excretory urography (at 1 and 6 months then annually). Mean follow-up was 35+/ 22.7 months. RESULTS: Mean operating time and mean length of hospital stay were 49.8+/-17.9min and four days (range: 2-10 days). Two cases of pyelonephritis were observed. The overall success was 70%. In the eight unresolved cases, the failure appeared at 2.7+/-1 month mean. Success criteria were moderate level of obstruction and primary junction. Here, patients so selected have 100% of success. CONCLUSION: Study confirmed retrograde laser endopyelotomy was safety with a short length of hospital stay. This minimally invasive procedure should be reserved to primary moderate stenoses without polar pedicle. PMID- 19135640 TI - [Retrocaval ureter based on a series of 16 cases]. AB - INTRODUCTION AND OBJECTIVES: To analyse the postoperative results of surgical treatment for retrocaval ureter. MATERIAL AND METHODS: The authors report a retrospective clinical study of a series of 16 patients (six women and 10 men) with a mean age of 38 years (range: 15-45 years) with retrocaval ureter treated between 1975 and 2005. The mean follow-up was 18 months (range: 12 to 34 months). All patients were evaluated by the standard diagnostic protocol for the time and were treated by one of the following surgical techniques: resection of the ureter and renal pelvis to renal pelvis anastomosis; resection of the ureter and pyelo ureteric anastomosis; resection of the ureter and oblique end-to-end uretero ureteric anastomosis; nephrectomy. RESULTS: The mean operating time was 95 min. Late postoperative follow-up revealed two cases (13%) of ureteric stenosis at the site of the oblique end-to-end uretero-ureteric anastomosis. Surgical revision was performed in one patient with resection of the ureter and reanastomosis, while anterograde dilatation of the stenosis was performed in the other patient. The postoperative course was uneventful in both patients. All patients were reviewed at six months with a satisfactory result, corresponding to reduction of hydronephrosis and improvement of renal function. CONCLUSION: Over the last 30 years, the diagnosis of retrocaval ureter has become more reliable and less invasive. Satisfactory results can be obtained with conventional surgical management. PMID- 19135641 TI - [Short or long schemes of antibiotic prophylaxis for prostate biopsy. A multicentre prospective randomised study]. AB - OBJECTIVE: We compared the incidence of the Urinary Tract Infection between a single preoperative dose and a three-day antibiotic prophylaxis regimen for transrectal ultrasound guided biopsy in randomized multicenter trial. MATERIAL AND METHODS: Between February 2006 and December 2007, 322 men who underwent transrectal ultrasound-guided prostate biopsy were included in a multicentre prospective randomised study. Patients received antibiotic prophylaxis by ciprofloxacin orally, either 1g single dose two hours before the biopsy (Group 1: n=139) or a prolonged prophylaxis for three days (Group 2: n=149). Assessment five days before and five days following the biopsy included a clinical examination, biological tests and a self-questionnaire. RESULTS: Two patients developed prostatitis, one in each group: 0.75% of the first group, 0.69% of the second. Twelve developed asymptomatic bacteriuria, six in each group: 4.51% of the first group and 4.19% of the second. There was no significant difference between the two groups (Fisher test; p>0.9). CONCLUSIONS: There was no significant difference between the two antibiotic prophylaxis regimen (one single dose or three days) for patients undergoing TRUS guided biopsies. Therefore, the single preoperative dose should be the preferred option. PMID- 19135642 TI - [Effects by androgen suppression with luteinizing hormone on cognitive functions in men treated for cancer of prostate]. AB - OBJECTIVE: To evaluate secondary effects on cognitive functions as memory and attention impairments in men treated by androgen suppression for cancer of prostate. MATERIAL AND METHODS: A prospective study on men treated with luteinizing hormone-realising hormone analogues (LH-RH analogues) for cancer of prostate was designed. Eighteen men were evaluated with cognitive tests as Mini Mental State (MMS), memory evaluation (Grober-Buschke, Rey Test), attention and executive test (Trail Making Test and Montgomery Asberg Depression Rating Scale [MADRS]) before beginning androgen suppression. Base line pretreatment assessments of every patient were compared with similar tests after six and 12 months of hormone therapy. RESULTS: On 18 patients, 14 were eligible with pretreatment-median PSA of 22ng/ml (13-659), testosterone of 477ng/dl (398-583), which decreased to 1 and 37ng/dl, respectively. Visual-memory test (Rey test) was significantly improved at M6 (p: 0,001) and M12 (p: 0,02) as was the inversed number-memory test (WAIS) after M6 of androgen suppression and (p: 0,03). CONCLUSION: After 12 months of androgen suppression, global cognitive performances were preserved. Our results failed to observe impairment of cognitive function. This deserves more important prospective study. PMID- 19135643 TI - [Tuna: clinical retrospective study addressing mid-term outcomes]. AB - PURPOSE: To estimate the efficiency and morbidity of the radiofrequency (Tuna) for treatment of symptomatic benign prostatic hyperplasia (BPH) in patients with medical treatment failure. MATERIAL AND METHODS: Between September 2003 and July 2007, 31 patients, mean age 59,5 years (50-76), were treated for BPH with Tuna. Patients had initially received medical treatment, which happened to fail and Tuna was offered as surgical treatment. Available clinical data of 28 patients was retrospectively analyzed. Mictionnel status was estimated with International Prostatic Symptoms Score (IPSS), urinary peak flow (UPF) and post void residual urine (PVR). Quality of life issues were also addressed. Efficiency of treatment was estimated on the evolution of these parameters after an average follow-up of 20 months (5-47). To estimate morbidity of treatment, peroperative and late complications were assessed. RESULTS: Twenty-eight patients were included in the study. At a mean follow-up of 20 months, a significant improvement of the mictionnel status was found for the IPSS, the quality of life, the Qmax (p<0,001) and the PVR (p<0,005) and quality of life. Morbidity was verified as minimal in the series. At a follow-up period of 20 months, 15 patients (53,6%) had stopped medical treatment, 10 patients (35,7%) pursued medical treatment and three patients (10,7%) had undergone another type of surgical treatment for BPH. The rate of re-treatment in the series was 46,4%. Among the group of patients without any medical treatment (15 patients), 10 patients reported satisfactory quality of life. CONCLUSION: Tuna is a technique with significant good results for the treatment of LUTS-BPH. The main advantage of this technique is its low morbidity, however, the rate of retreatment was observed as high. Series of patients with long-term follow-up should provide evidence to objectively define the role of Tuna in BPH surgical treatment. PMID- 19135644 TI - [What is the publication rate of papers presented at the French Association of Urology annual conferences?]. AB - The meeting of the French Association of Urology (AFU) occurs once a year. Some communications are consequently published in national or international journals. We analysed the criteria of publication of works communicated during the AFU meeting. MATERIAL: We analysed all the communications of the 94th and the 95th meeting of AFU from years 2000 and 2001, identified from the Progres en urologie supplements. We then asked Medline covering a period of time from January 1996 to December 2006 and then compared communications to similar published articles. RESULTS: The overall publication rate was 34.5%. Articles (38.6%) were published the year following the meeting but the main articles were published the following 2 years after the meeting (78.42%). The mean time for publication was 16.9months. The prospective studies had a publication rate of 44.1% versus 24.8% for retrospective studies. One article over five was published in Progres en urologie (21.6%). CONCLUSIONS: Only one third of communications of the French meeting of urology was then published. Twenty percent were published in Progres en urologie. Different way could increase the publication rate. The time for publication, higher for this meeting than for others, may decrease with the new submission website. PMID- 19135645 TI - [Scientific work: oral presentation or publication?]. PMID- 19135646 TI - [Primary carcinoma of the head of the epididymis]. AB - The authors report the case of a 44-year-old man in whom a poorly differentiated primary carcinoma of the head of the epididymis was discovered incidentally. Due to the rarity of this diagnosis, a comprehensive assessment was performed looking for a primary tumour, but without success. Despite early surgical resection, the patient developed lymph-node metastases. This exceptional tumour showed low sensitivity to chemotherapy. Malignant tumours of the epididymis are exceptional and require investigations to detect a primary tumour. Treatment is based on surgical resection, ideally via an inguinal incision, combined with chemotherapy adapted to the histological type. PMID- 19135647 TI - Normal people working in normal organizations with normal equipment: system safety and cognition in a mid-air collision. AB - A fundamental challenge in improving the safety of complex systems is to understand how accidents emerge in normal working situations, with equipment functioning normally in normally structured organizations. We present a field study of the en route mid-air collision between a commercial carrier and an executive jet, in the clear afternoon Amazon sky in which 154 people lost their lives, that illustrates one response to this challenge. Our focus was on how and why the several safety barriers of a well structured air traffic system melted down enabling the occurrence of this tragedy, without any catastrophic component failure, and in a situation where everything was functioning normally. We identify strong consistencies and feedbacks regarding factors of system day-to day functioning that made monitoring and awareness difficult, and the cognitive strategies that operators have developed to deal with overall system behavior. These findings emphasize the active problem-solving behavior needed in air traffic control work, and highlight how the day-to-day functioning of the system can jeopardize such behavior. An immediate consequence is that safety managers and engineers should review their traditional safety approach and accident models based on equipment failure probability, linear combinations of failures, rules and procedures, and human errors, to deal with complex patterns of coincidence possibilities, unexpected links, resonance among system functions and activities, and system cognition. PMID- 19135648 TI - Social anxiety and emotional suppression: the mediating role of beliefs. AB - There is mounting evidence to suggest that social anxiety is associated with the suppression of emotional expression. The current study examined self-reported emotional suppression and beliefs about expressing emotions among undergraduate students (n=95). Socially anxious undergraduates reported greater use of emotional suppression compared to their non-socially anxious peers. They also reported greater ambivalence about emotional expression, more difficulties in emotional responding, more fears of emotional experiences, and more negative beliefs about emotional expression. Believing that emotional expression must be kept in control and is a sign of weakness partially mediated the association between social anxiety and emotional suppression. PMID- 19135649 TI - Distress tolerance and early adolescent externalizing and internalizing symptoms: the moderating role of gender and ethnicity. AB - A large body of research has examined the development of internalizing and externalizing symptoms in childhood and early adolescence. Notably, there is significant concomitant impairment associated with early adolescent symptomatology, as well as association of these symptoms with future development of psychopathology, poor physical health, self-destructive thoughts and behaviors, criminal behavior, and HIV risk behaviors. Drawing on negative reinforcement theory, the current study sought to examine the potential role of distress tolerance, defined as the ability to persist in goal-directed activity while experiencing emotional distress, as a potential mechanism that may underlie both internalizing and externalizing symptoms among 231 Caucasian and African American youth (M age=10.9 years; 45.5% female; 54.5% Caucasian ethnicity). A series of regressions resulted in significant moderated relationships, such that low distress tolerance conferred increased risk for alcohol use among Caucasians, delinquent behavior among African Americans, and internalizing symptoms among females. Clinical implications, including the potential role of negative reinforcement models in early intervention with young adolescents, are discussed. PMID- 19135650 TI - Effects of verbal information on fear-related reasoning biases in children. AB - The present study made an attempt to induce fear-related reasoning biases by providing children with negative information about a novel stimulus. For this purpose, non-clinical children aged 9-12 years (N=318) were shown a picture of an unknown animal for which they received either negative, ambiguous, positive, or no information. Then children completed a series of tests for measuring various types of reasoning biases (i.e., confirmation bias and covariation bias) in relation to this animal. Results indicated that children in the negative and, to a lesser extent, the ambiguous information groups displayed higher scores on tests of fear-related reasoning biases than children in the positive and no information groups. Altogether, these results support the idea that learning via negatively tinted information plays a role in the development of fear-related cognitive distortions in youths. PMID- 19135651 TI - Differential allelic expression of dopamine D1 receptor gene (DRD1) is modulated by microRNA miR-504. AB - BACKGROUND: Previously, we reported that dopamine D1 receptor gene (DRD1) is associated with nicotine dependence (ND) and demonstrated that two alleles (A and G) of polymorphism rs686 in the 3'-untranslated region (3'UTR) of DRD1 are expressed differentially. However, the mechanism underlying the differential expression remains to be determined. We hypothesize that it is caused by miRNA targeting. METHODS: We first used the MicroInspector algorithm to identify microRNAs (miRNAs) potentially targeting the rs686 polymorphism in the DRD1 3'UTR and then employed a luciferase reporter assay combined with site-directed mutagenesis to test the predicted miRNA targeting. We also examined the miRNA targeting of DRD1 with a gene expression assay. RESULTS: Of two miRNAs predicted by computational analyses, we found that miR-504, not miR-296, upregulated reporter luciferase activity and increased DRD1 expression by targeting the DRD1 3' UTR, whereas inhibition of miR-504, not miR-296, had the opposite effect. Furthermore, we showed that the direct binding of miR-504 to the DRD1 3'UTR, verified by site-directed mutagenesis, causes a significant expression difference between the two alleles. CONCLUSIONS: miR-504 up-regulates DRD1 expression by direct binding to the 3'UTR, which leads to differential allele-specific expression of DRD1. PMID- 19135653 TI - Connectionist semantic systematicity. AB - Fodor and Pylyshyn [Fodor, J. A., & Pylyshyn, Z. W. (1988). Connectionism and cognitive architecture: A critical analysis. Cognition, 28, 3-71] argue that connectionist models are not able to display systematicity other than by implementing a classical symbol system. This claim entails that connectionism cannot compete with the classical approach as an alternative architectural framework for human cognition. We present a connectionist model of sentence comprehension that does not implement a symbol system yet behaves systematically. It consists in a recurrent neural network that maps sentences describing situations in a microworld, onto representations of these situations. After being trained on particular sentence-situation pairs, the model can comprehend new sentences, even if these describe new situations. We argue that this systematicity arises robustly and in a psychologically plausible manner because it depends on structure inherent in the world. PMID- 19135652 TI - Circadian alteration in neurobiology during 30 days of abstinence in heroin users. AB - BACKGROUND: Previous studies have shown that individuals withdrawn from chronic opiate administration undergo substantial elevations of cortisol levels with blunted corticotropin (ACTH) rhythms and that these changes persist beyond the 7 10 days of acute withdrawal symptoms. However, there are no published studies of changes in expression of clock genes or of other neuropeptides related to circadian-rhythm regulation, which may influence relapse susceptibility. METHODS: Blood samples were collected from 8 healthy control subjects and 16 heroin addicts during pharmacologically unassisted withdrawal on the 3rd, 10th, and 30th days of abstinence at 3-hour intervals for 24 hours. Outcome measures were the relative expression of clock gene mRNA (hperiod1, hperiod2, hclock) and the levels of serum cortisol, plasma ACTH, beta-endorphin (beta-EP), leptin, neuropeptide Y, interleukin-2 (IL-2), and tumor necrosis factor (TNF) in these subjects. RESULTS: Compared with healthy volunteers, abstinent addicts showed disruptions in diurnal rhythms of hPER1 and hPER2 mRNA expression, along with disruptions in diurnal rhythms of cortisol, ACTH, beta-endorphin, leptin, and IL 2 release. Several of these disruptions (hPER1, hPER2, ACTH, beta-endorphin, and IL-2) persisted for the 30-day testing period, as did elevation of 24-hour levels of cortisol and decreases in 24-hour IL-2 and TNF levels. CONCLUSIONS: These prolonged neurobiological changes may play a role in protracted opiate withdrawal symptoms and contribute to relapse vulnerability. PMID- 19135654 TI - Beyond perceptual symbols: a call for representational pluralism. AB - Recent evidence from cognitive neuroscience suggests that certain cognitive processes employ perceptual representations. Inspired by this evidence, a few researchers have proposed that cognition is inherently perceptual. They have developed an innovative theoretical approach that rests on the notion of perceptual simulation and marshaled several general arguments supporting the centrality of perceptual representations to concepts. In this article, I identify a number of weaknesses in these arguments and defend a multiple semantic code approach that posits both perceptual and non-perceptual representations. PMID- 19135655 TI - Does history repeat itself? Cortical columns 1. Introduction. PMID- 19135656 TI - An investigation of the visual disturbances experienced by patients on clomiphene citrate. AB - OBJECTIVE: To evaluate the impact of clomiphene citrate on vision. DESIGN: Observational study. SETTING: Patients were referred to the University of Ottawa Eye Institute ophthalmology clinic from the Department of Obstetrics and Gynaecology of the Ottawa Hospital-General Campus. PATIENT(S): Eight adult females taking clomiphene citrate and experiencing visual disturbances. INTERVENTION(S): Patients received a comprehensive visual evaluation twice: once during a washout period, and once during an active clomiphene citrate treatment. MAIN OUTCOME MEASURE(S): Ophthalmologic examination, color vision, visual acuity, contrast sensitivity, visual fields using standard automated perimetry, and foveal flicker sensitivity at high (32 Hz) and low (8 Hz) temporal frequencies. RESULT(S): We found no differences between the washout and clomiphene citrate conditions for color vision, visual acuity, contrast sensitivity, and visual fields. The only statistically significant difference was found for foveal flicker sensitivity at 32 Hz in the right eye, with a similar trend in the left eye and at 8 Hz in both eyes. CONCLUSION(S): The effect of clomiphene citrate on vision was minimal, and the visual disturbances were reversible in all patients. A bilateral reduction in flicker sensitivity was the only observed visual disturbance. Women who experience visual symptoms associated with clomiphene citrate should be monitored, but therapy can usually be maintained. PMID- 19135657 TI - A randomized, controlled clinical trial comparing the effects of aromatase inhibitor (letrozole) and gonadotropin-releasing hormone agonist (triptorelin) on uterine leiomyoma volume and hormonal status. AB - OBJECTIVE: To examine and compare the efficacy and safety of GnRH agonist (GnRHa) vs. aromatase inhibitor in premenopausal women with leiomyomas. DESIGN: Multicenter, randomized, controlled clinical trial. SETTING: University hospitals. PATIENT(S): A total of 70 subjects with a single uterine myoma measuring >or=5 cm. Subjects were randomized into two groups with use of a random table. They were treated with aromatase inhibitor (group A) or GnRHa (group B). INTERVENTION(S): Group A received letrozole (2.5 mg/d) for 12 weeks. Group B received triptorelin (3.75 mg/mo) for 12 weeks. MAIN OUTCOME MEASURE(S): Measurement of myoma volume and E(2), FSH, LH, and T levels. RESULT(S): Total myoma volume decreased by 45.6% in group A and 33.2% in group B. Reductions in myoma volume in the two groups were statistically significant. There was no significant change in hormonal milieu in group A. The serum level of hormones significantly decreased in group B by the 12th week of treatment. CONCLUSION(S): Uterine myoma volume was successfully reduced by use of an aromatase inhibitor. Rapid onset of action and avoidance of initial gonadotropin flare with an aromatase inhibitor may be advantageous for short-term management of women with myomas of any size who are to be managed transiently and who wish to avoid surgical intervention, specifically women with unexplained infertility having uterine myoma. PMID- 19135658 TI - Translocation chromosome karyotypes of the Robertsonian translocation carriers' embryos. AB - OBJECTIVE: To explore the translocation karyotypes of the Robertsonian translocation (RT) carriers' embryos in their preimplantation genetic diagnosis (PGD) cycles. DESIGN: Retrospective. SETTING: University-affiliated IVF center. PATIENT(S): A total of 35 RT carrier couples underwent 39 blastomere PGD cycles from August 2005 to June 2008. INTERVENTION(S): The PGD analysis of embryos. MAIN OUTCOME MEASURE(S): Meiotic segregation patterns of the 253 embryos were analyzed in their PGD cycles. RESULT(S): The alternate embryos were approximately one third of all the embryos (82 of 253, 32.41%). The ratio among the alternate embryos, the adjacent-1 embryos and the adjacent-2 embryos was 2:1:1. There was no significant difference between the male RT subgroup and the female RT subgroup (31.43% vs. 37.21%) regarding the proportion of the alternate embryos and no significant difference was found between the common RT subgroup and the rare RT subgroup (33.48% vs. 25.00%). In contrast, the alternate embryo percentage in the pregnancy subgroup was much higher than that of the pregnancy failure subgroup (45.16% vs. 28.27%). CONCLUSION(S): The alternate embryos are dominant among the RT carriers' embryos and different gender and different translocation chromosomes have no effect on the alternate embryos rate. The ratio among the alternate embryos, the adjacent-1 embryos and the adjacent-2 embryos is 2:1:1. PMID- 19135659 TI - Outcome of twin babies free of Von Hippel-Lindau disease after a double-factor preimplantation genetic diagnosis: monogenetic mutation analysis and comprehensive aneuploidy screening. AB - OBJECTIVE: To increase the embryo implantation rate, a double-factor preimplantation genetic diagnosis (DF-PGD) was performed, selecting for transfer potentially euploid evolved embryos free of the mutation responsible for Von Hippel-Lindau syndrome (VHL). DESIGN: Case report. SETTINGS: Medical university center and a private IVF center. PATIENT(S): A patient carrier of the R161Q mutation on the VHL gene. INTERVENTION(S): After first polar body (1PB) biopsy, it was analyzed using comparative genomic hybridization (1PB-CGH). On day +3, mutation detection using minisequencing and short tandem repeat analysis was performed in multiple displacement amplification products of a single blastomere per embryo. MAIN OUTCOME MEASURE(S): Transfering embryos free of the disease and originating from euploid oocytes. RESULT(S): Nine of the twelve oocytes obtained were successfully analyzed using 1PB-CGH. One of them was aneuploid (1PB #1: 29XX,+2,+10,+12,+17,+19), and the rest were euploid. All of the oocytes were fertilized and became evolved embryos. Six of the embryos were VHL unaffected and had good quality. Five (83%) of them were potentially euploid. According to cytogenetic results, two of the evolved and healthy embryos were transfered, achieving the birth of healthy twin babies. CONCLUSION(S): The DF-PGD can be a useful tool to increase implantation of transfered embryos in PGD for monogenic diseases. PMID- 19135660 TI - A case of a methotrexate-resistant ectopic pregnancy in which dactinomycin was effective as a second-line chemotherapy. AB - OBJECTIVE: To report a case of a methotrexate (MTX)-resistant heterotopic interstitial pregnancy in which dactinomycin was necessary as a second-line chemotherapy. DESIGN: Case report. SETTING: Medical college-affiliated hospital. PATIENT(S): A 39-year-old woman with vaginal bleeding after the transfer of two thawed 8-cell embryos. INTERVENTION(S): MTX 50 mg/m(2) was given to the patient to treat a concomitant pregnancy in which an interstitial pregnancy and an intrauterine blighted ovum were identified. However, the patient's hCG level rose despite two MTX administrations. As a result, dactinomycin was then administered as a second-line chemotherapy. The patient's serum hCG level declined to a normal range after two administrations of dactinomycin. CONCLUSION(S): Dactinomycin could be an alternative in cases for which MTX is not effective and persistently high hCG levels exist. PMID- 19135661 TI - Sperm chromosome analysis of an infertile patient with a 95% mosaic r(21) karyotype and normal phenotype. AB - OBJECTIVE: To examine sperm meiotic segregation in a man with mosaic ring chromosome 21. DESIGN: Case report. SETTING: Hospital departments of reproductive biology, cytogenetics, gynecology, and obstetrics. PATIENT(S): One patient referred for cryptozoospermia, heterozygous for a ring chromosome. INTERVENTION(S): Fluorescence in situ hybridization with chromosome 21-specific probes after sperm selection. RESULT(S): A total of 169 spermatozoa were selected; 92.3% carried a normal 21 chromosome, 6.5% the ring chromosome, and 1.2% both. CONCLUSION(S): Ring chromosome frequency in mature sperm cells was low and may be due to preferential meiosis of normal spermatogonia,which could explain the cryptozoospermia and unexpected ratio in this case. PMID- 19135662 TI - Attitudes and intention to donate oocytes for research. AB - OBJECTIVE: In 2007, the Human Fertilization and Embryology Authority permitted oocyte donation for research through voluntary donation or within an oocyte share model. The aims of this study were to investigate volunteer (nonpatient) women's attitudes and intentions to donate using components of the Theory of Planned Behavior and their attitudes toward parenthood through structural equation modeling. DESIGN: Questionnaires. SETTING: Online. PATIENT(S): A total of 253 nonpatient women. MAIN OUTCOME MEASURE(S): Attitudes towards oocyte donation for research and reasons for parenthood scale. RESULT(S): Of the 253 respondents, 94 were potential donors, 98 were possible donors, and 61 were non-donors. Most potential donors (68%) reported no preference towards donating their oocytes for research or an infertile couple. Structural equation modeling revealed that age (beta = -.03) and components of the TPB (beta = .16) had a statistically significant direct effect on intentions to donate for research. Attitudes toward parenthood was not linked to intentions to donate for research. CONCLUSION(S): There appears to be a strong altruistic motive along with the theoretical underpinnings of positive attitudes, feeling supported, and accepting the consequences of oocyte donation for research, suggesting these have the potential to inform recruitment practices and tailor clinical services. PMID- 19135663 TI - Genetic analysis of human embryos by metaphase comparative genomic hybridization (mCGH) improves efficiency of IVF by increasing embryo implantation rate and reducing multiple pregnancies and spontaneous miscarriages. AB - OBJECTIVE: To assess the benefit of selecting blastocysts for cryotransfer based upon prior comparative genomic hybridization (CGH) karyotyping of blastomeres derived from their cleaved embryos of origin. Implantation and birth rates per transfer of previtrified CGH-tested blastocysts were compared with those following the transfer of nonCGH-tested fresh and warmed embryos. DESIGN: In vitro studies. SETTING: Private infertility clinic. PATIENT(S): Women undergoing infertility treatment. INTERVENTION(S): Three groups of women with similar clinical and demographic characteristics were compared. Group A underwent transfer of warmed blastocysts derived from CGH-normal day 3 embryos. Group B underwent embryo transfer of warmed blastocysts derived from nonkaryotyped vitrified embryos. Group C underwent fresh transfers with non-CGH-tested blastocysts. MAIN OUTCOME MEASURE(S): Implantation and birth rates per embryo after the cryotransfer of CGH-tested blastocysts. RESULT(S): The birth rate per transferred blastocyst in group A was 48%, versus 15% for group B and 19% for group C. The birth rate per embryo transfer was 60% for group A, and 33% for group B and 36% for group C. The miscarriage rate was 4% in group A, 8% in group B, and 12% in group C. CONCLUSION(S): The transfer of previously vitrified blastocysts derived from CGH-normal embryos significantly improves implantation and birth rates per embryo transferred and reduces the miscarriage rate. Vitrification does not compromise this enhancement. PMID- 19135665 TI - The effect of repeated freezing and thawing on human sperm DNA fragmentation. AB - OBJECTIVE: To investigate the effects of repeated freezing and thawing on human sperm motility, vitality, and DNA integrity. DESIGN: A prospective clinical study. SETTING: Tertiary care fertility clinic. PATIENT(S): Twenty men presenting for infertility investigations. INTERVENTION(S): Each sample was subjected to three cycles of freezing and thawing both with and without washing steps and the addition of fresh cryoprotectant between each cycle. MAIN OUTCOME MEASURE(S): Percentage sperm DNA fragmentation, motility, and vitality before and following repeated freezing and thawing. RESULT(S): The percentage sperm DNA fragmentation rose significantly following each freeze-thaw cycle; however, samples that were not washed and to which fresh cryoprotectant was not added after each cycle fared significantly better than their washed counterparts. Both motility and vitality decreased steadily following each cycle but cell survival was significantly greater in the unwashed samples. CONCLUSION(S): In terms of the level of sperm DNA fragmentation, up to three cycles of freezing and thawing can be performed with a level of risk comparable to that following a single cycle of freezing and thawing. This is provided that samples are refrozen in their original cryoprotectant and not washed or altered in any way in between, and provided that they are separated by density gradient centrifugation or swim-up before use in assisted reproduction technology. PMID- 19135664 TI - The antifibrotic drug halofuginone inhibits proliferation and collagen production by human leiomyoma and myometrial smooth muscle cells. AB - OBJECTIVE: To investigate the effects of the antifibrotic drug halofuginone on extracellular matrix production, cell proliferation, and apoptosis of cultured myometrial and leiomyoma smooth muscle cells. DESIGN: Comparative and controlled experimental research study. SETTING: University research laboratory. PATIENT(S): Leiomyoma and myometrial tissues were obtained from eight different patients at the time of elective hysterectomy. MAIN OUTCOME MEASURE(S): The effects of halofuginone on cell proliferation were assessed by tritiated thymidine uptake assays and cell count assays. Effects on TGFbeta1, collagen type I, and collagen type III mRNA levels were assessed by quantitative real-time polymerase chain reaction. Effects on apoptosis were assayed using a chemiluminescent assay to measure changes in caspase 3 and 7. RESULT(S): Halofuginone inhibited cell proliferation of both leiomyoma and autologous myometrial cells in a dose dependent manner by inhibiting DNA synthesis within 24 hours and later inducing apoptosis (as measured by increased caspase 3/7) by 48-72 hours. Halofuginone also significantly reduced collagen type I (alpha1) and collagen type III (alpha1) mRNA levels, as well as the profibrotic factor TGFbeta1 mRNA levels in both cell types. CONCLUSION(S): These results provide evidence to support the use of the antifibrotic drug halofuginone as a novel drug treatment for uterine leiomyomas. PMID- 19135666 TI - Prospective cohort study of three- versus two-dimensional ultrasound for prediction of oocyte maturity. AB - OBJECTIVE: To determine whether three-dimensional follicular ultrasound (3-D) measurements are better predictors of optimal timing of hCG administration than two-dimensional (2-D) images, the current standard. DESIGN: Prospective, cohort study. SETTING: Tertiary referral center. PATIENT(S): Seventy-six patients undergoing IVF. INTERVENTION(S): Seventy-six consecutive patients undergoing serial follicular monitoring during IVF had an additional daily 3-D volume scan of their ovaries once lead follicles had reached 16 mm diameter. MAIN OUTCOME MEASURE(S): Number of mature oocytes retrieved. RESULT(S): The 2-D follicular diameter measurements predicted 25.4% of the observed variance in the number of mature oocytes retrieved. The 3-D follicular volume measurements were more predictive of outcome, accounting for 29.2% of the observed variance in number of mature oocytes retrieved. Follicles >22 mm diameter and 5 mL volume were associated with fewer mature oocytes reflecting an undesired postmature state. Follicles measuring 11 to 15 mm had a 50% chance of yielding a mature oocyte. CONCLUSION(S): Three-dimensional follicular volume measurements have a stronger correlation with the number of mature oocytes retrieved than 2-D measurements. As 3-D technology improves, this parameter may replace 2-D measurements in the optimal timing of hCG before oocyte retrieval. PMID- 19135667 TI - Prevalence of polycystic ovary syndrome in young women who had idiopathic central precocious puberty. AB - OBJECTIVE: To assess the prevalence of polycystic ovary syndrome (PCOS) in a cohort of young women with previous idiopathic central precocious puberty (ICPP) at least 3 years after menarche, and to look for any predictive factors of PCOS at the time ICPP was diagnosed. DESIGN: Longitudinal study. SETTING: Pediatrics unit, Verona, Italy. PATIENT(S): Forty-six young women (18.1 +/- 3.0 years) who had been treated with GnRH analogues during childhood, observed at gynecologic age of 6.23 +/- 3.3 years. INTERVENTION(S): Semistructured interview concerning cycles, physical exam, blood sampling, and transabdominal pelvic ultrasound. MAIN OUTCOME MEASURE(S): Oligomenorrhea, LH, FSH, E(2), T, DHEAS, free T, delta4 androstenedione, 17-OHP, P, polycystic ovary morphology (PCOM). RESULT(S): Fifteen percent of the young women had oligomenorrhea, 28% clinical hyperandrogenism, 48% biochemical hyperandrogenism, and 37% PCOM. A total of 32% of the patients had PCOS according to the Rotterdam definition and 30% had PCOS according to the Androgen Exess Society. The prevalent phenotype of PCOS was characterized by clinical and/or biochemical hyperandrogenism and PCOM. We did not find any predictive factors for PCOS at the time ICPP was diagnosed. CONCLUSION(S): Patients with ICCP are prone to developing PCOS. The prominent phenotype in this cohort was PCOM associated with clinical and/or biochemical hyperandrogenism. Further follow-ups of these young adult patients will clarify whether this phenotype persists and if it will have important long-term implications regarding increased risk of infertility or metabolic complications. PMID- 19135668 TI - Up-regulation of endocrine gland-derived vascular endothelial growth factor but not vascular endothelial growth factor in human ectopic endometriotic tissue. AB - OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF), endocrine gland-derived VEGF (EG-VEGF/PK1), and its receptors (PKR1 and PKR2) in eutopic and ectopic endometrial tissues. DESIGN: A case-control study. SETTING: University reproduction unit. PATIENT(S): Infertile women undergoing diagnostic laparoscopy for tubal patency. INTERVENTION(S): Endometrial and endometriotic tissue sampling from women with and without endometriosis. MAIN OUTCOME MEASURE(S): Quantitative polymerase chain reaction (PCR) analysis of genes in eutopic and ectopic endometrial tissues. The EG-VEGF protein was studied by immunohistochemistry. RESULT(S): In normal endometrium, EG-VEGF messenger RNA (mRNA) expression was 50-fold higher in the secretory than in the proliferative phase, but that of PKR1 was 6-fold higher in the latter than in the former. The PKR2 transcript was detected in the proliferative but not the secretory endometrium. In patients with endometriosis, eutopic endometrial PKR2 transcript level was 4-fold higher in the proliferative than in the secretory phase. No differences in EG-VEGF or PKR1 were found in proliferative versus secretory endometrium in these patients. There were no significant differences in the expression of EG-VEGF in eutopic endometrium of normal women and in those with endometriosis. In the paired laser-captured microdissected eutopic endometrial and ectopic endometriotic samples, a significantly higher EG-VEGF, but not VEGF, transcript level was detected in the ectopic when compared with eutopic samples; whereas the expressions of PKR1 and PKR2 were barely detectable. The H-scoring confirmed that the stroma of endometriotic samples had a significantly higher EG VEGF protein expression than that in the paired eutopic endometrium. CONCLUSION(S): High levels of EG-VEGF expression may play an important role in angiogenesis in endometriotic tissues. PMID- 19135669 TI - FIGO staging for uterine sarcomas. PMID- 19135670 TI - Circulating TNFalpha levels in older men and women do not show independent prospective relations with MI or stroke. AB - BACKGROUND: Tumour necrosis factor alpha (TNFalpha) is a pro-inflammatory cytokine implicated in atherosclerotic plaque formation. We investigated whether circulating TNFalpha is prospectively associated with myocardial infarction (MI) or stroke in the older general population, independently of established cardiovascular risk factors and other inflammatory markers related to CHD risk. METHODS: We measured baseline TNFalpha concentrations in stored serum samples of 362 incident MI and 299 incident stroke cases and controls (2 per case, frequency matched by age, gender and town) who were 'nested' in parallel prospective studies of 4252 men and 4286 women aged 60-79 years assessed in general practices in 24 British towns in 1998-2000 and followed up for an average 7 years for fatal and non-fatal MI and stroke. RESULTS: TNFalpha levels were 11.4% (95% CI 9.5, 13.3%) higher among MI cases than controls; geometric mean 1.84 pg/mL compared to 1.63 pg/mL, p (difference)<0.001. Participants in the top third of baseline TNFalpha levels had an age-adjusted odds ratio (OR) for MI of 1.75 (95%CI 1.22, 2.49) compared with those in the bottom third, which was reduced to 1.47 (95%CI 1.01, 2.14) after adjustment for established cardiovascular risk factors. However, further adjustment for C-reactive protein and interleukin-6 abolished the association OR 1.33 (95% CI 0.91, 1.66) and the linear trend. Excluding subjects with pre-existing CVD did not materially affect results. No significant association between TNFalpha and stroke was observed. CONCLUSIONS: This study suggests that TNFalpha is not a strong independent risk marker for MI, and is not associated with risk of stroke. PMID- 19135671 TI - Pioglitazone vs glimepiride: Differential effects on vascular endothelial function in patients with type 2 diabetes. AB - OBJECTIVE: The aim of this study was to compare the effect of glimepiride and pioglitazone on endothelial function in patients with type 2 diabetes already on metformin. METHODS: Twenty-eight patients with type 2 diabetes already on metformin, without known cardiovascular disease, were randomized in 2 groups; glimepiride (4 mg od) was added in group A (n=14) and pioglitazone (30 mg od) in group B (n=14) for 6 months. Flow-mediated dilation (FMD) in the brachial artery was assessed in all patients, at baseline and at follow-up. RESULTS: The 2 groups did not differ in age (mean+/-S.D., 63.6+/-7.3 years vs 62.8+/-7.2 years respectively), or any measured variable at baseline. Fasting glucose and glycated haemoglobin improved similarly in both groups. There were significant differences between the 2 groups in the absolute changes observed at follow-up in waist circumference, +1.86+/-3.11 cm vs -1.86+/-1.88 cm in groups A and B respectively; fasting insulin levels, +14.79+/-12.56 pmol/L vs -25.84+/-28.09 pmol/L; homeostasis model assessment (HOMA), +0.66+/-1.01 vs -1.83+/-1.38; HDL cholesterol levels, -0.07+/-0.22 mmol/L vs +0.14+/-0.20 mmol/L and FMD, +0.14+/ 1.09% vs +2.02+/-2.05% (p<0.05 for all). The only independent predictor factor of the FMD improvement was treatment-induced changes in HOMA (R(2): 0.488, slope: 0.782, [95% CI: -1.128, -0.436], p=0.0001). CONCLUSIONS: In patients with type 2 diabetes already on metformin, addition of pioglitazone as compared to glimepiride, improved endothelial function despite similar glycemic control. The improvement in endothelial function was mainly due to a reduction in insulin resistance. PMID- 19135673 TI - Our primary effort was to increase the internationalisation of the symposium, which was born and--until now--was always held in Europe. Foreword. PMID- 19135672 TI - A novel 3-dimensional micro-ultrasound approach to automated measurement of carotid arterial plaque volume as a biomarker for experimental atherosclerosis. AB - Improved methods for non-invasive in vivo assessment are needed to guide development of animal models of atherosclerosis and to evaluate target engagement and in vivo efficacy of new drugs. Using novel 3D-micro-ultrasound technology, we developed and validated a novel protocol for 3D acquisition and analysis of imaging to follow lesion progression in atherosclerotic mice. The carotid arteries of ApoE receptor knockout mice and normal control mice were imaged within the proximal 2mm from the aortic branch point. Plaque volume along that length was quantified using a semi-automated 3D segmentation algorithm. Volumes derived by this method were compared to those calculated using 3-D histology post mortem. Bland-Altman comparison revealed close correlation between these two measures of plaque volume. Furthermore, using a segmentation technique that captures early positive and 33 week negative remodeling, we found evidence that plaque volume increases linearly over time. Each animal and each plaque served as its own control, allowing accurate comparison. The high fidelity anatomical registration of this protocol provides increased spatial resolution and therefore greater sensitivity for measurement of plaque wall size, an advance over 2 dimensional measures of intimal-medial-thickening. Further, 3-dimensional analysis ensures a point of registration that captures functional markers in addition to the standard structural markers that characterize experimental atherosclerosis. In conclusion, this novel imaging protocol provides a non invasive, accurate surrogate marker for experimental atherosclerosis over the life of the entire lesion. PMID- 19135674 TI - Aqueous normal-phase retention of nucleotides on silica hydride columns. AB - The use of silica hydride-based stationary phases for the retention and analysis of nucleotides has been investigated. Both reversed-phase columns with a hydride surface underneath as well as those with an unmodified or a minimally modified hydride material were tested. With these systems, an aqueous normal-phase mode was used with high organic content mobile phases in combination with an additive to control pH for the retention of the hydrophilic nucleotides. Isocratic and gradient elution formats have been used to optimize separations for mixtures containing up to seven components. All conditions developed are suitable for methods that utilize mass spectrometry detection. PMID- 19135675 TI - Simultaneous measurement of proline and related compounds in oak leaves by high performance ligand-exchange chromatography and electrospray ionization mass spectrometry for environmental stress studies. AB - A mass spectrometer was coupled to high-performance ligand-exchange liquid chromatography (HPLEC) for simultaneous analysis of stress associated solutes such as proline, hydroxyproline, methylproline, glycine betaine and trigonelline extracted from leaves of drought stressed oaks and an internal standard namely N acetylproline. Methanol/chloroform/water extracts were analyzed using an Aminex HPX-87C column and specifically quantified by the positive ion mode of an electrospray ionisation-mass spectrometry (ESI-MS) in single ion monitoring (SIM) mode. The recovery of N-acetyl proline added to oak leaf extracts ranged from 85.2 to 122.1% for an intra-day study. Standard calibration curves showed good linearity in the measured range from 0.3125 to 10micromolL(-1) with the lowest correlation coefficient of 0.99961 for trigonelline. The advantages of this alternative procedure, compared to previously published methods using fluorescence or amperometric detections, are the simultaneous and direct detection of osmoprotectants in a single chromatographic run, a minimal sample preparation, a good specificity and reduced limits of quantification, ranging from 0.1 to 0.6micromolL(-1). Fifty-six days of water deficit exposure resulted in increased foliar free proline levels (2.4-fold, P<0.001, 155micromolg(-1) FW) and glycine betaine contents (2.5-fold, P<0.05, 175micromolg(-1) FW) of drought stressed oak compared to control. PMID- 19135676 TI - GUESSmix-guided optimization of elution-extrusion counter-current separations. AB - Rational strategies for the optimization of separations are vital to any chromatographic technique. In counter-current separations (CS), once a suitable solvent system is selected for a given separation, the operator is faced with the task of optimizing the separation through the manipulation of those adjustable operation parameters allowed for by the current CS technology. This study employed a mixture of 21 natural products of varying polarity, molecular mass, and functionality, termed the GUESSmix, as a tool to assess the effectiveness of optimization strategies. The behavior of the GUESSmix was observed in the hexane/ethyl acetate/methanol/water 4:6:4:6 (HEMWat +3) solvent system. The effect of operation parameters on both the elution and extrusion stages of a recently introduced CS methodology, termed elution-extrusion counter-current chromatography (EECCC), was investigated. The resulting chromatograms were plotted with K-based reciprocal symmetry plots (ReS and ReSS), which allow comparison of the K values of significant peaks and assessment of resolution of eluting compounds in the interval 0< or =K< or =infinity. The operation parameters studied were: (1) the effect of temperature controlled water circulation around the centrifuge; (2) the combination of flow rate and revolution speed; (3) sample loading capacity; (4) the direction of rotation either agreeing with or opposing the direction of coil winding; (5) injection before equilibration, a practice that saves operator time and reduces solvent consumption. The GUESSmix was found to be a highly useful reference mixture to compare and contrast stationary phase retention volume ratios, resolution, K values, peak shapes, and extrusion characteristics between CS experiments. EECCC is shown to be a robust technique that may be enhanced with appropriate temperature, rpm, flow rate, sample loading, direction of rotation, and injection timing. Plotting ReS[S] chromatograms enables systematic study of CS parameters, which are not reproducibly represented in retention time or volume plots. PMID- 19135677 TI - Drop-to-drop microextraction across a supported liquid membrane by an electrical field under stagnant conditions. AB - Electromembrane extraction (EME) of basic drugs from 10 microL sample volumes was performed through an organic solvent (2-nitrophenyl octyl ether) immobilized as a supported liquid membrane (SLM) in the pores of a flat polypropylene membrane (25 microm thickness), and into 10 microL 10 mM HCl as the acceptor solution. The driving force for the extractions was 3-20 V d.c. potential sustained over the SLM. The influence of the membrane thickness, extraction time, and voltage was investigated, and a theory for the extraction kinetics is proposed. Pethidine, nortriptyline, methadone, haloperidol, and loperamide were extracted from pure water samples with recoveries ranging between 33% and 47% after only 5 min of operation under totally stagnant conditions. The extraction system was compatible with human urine and plasma samples and provided very efficient sample pretreatment, as acidic, neutral, and polar substances with no distribution into the organic SLM were not extracted across the membrane. Evaluation was performed for human urine, providing linearity in the range 1-20 microg/mL, and repeatability (RSD) in average within 12%. PMID- 19135678 TI - Ultra-high concentration of amylose for chiral separations in capillary electrophoresis. AB - In the present study, a capillary electrophoresis method using high concentration of amylose solutions as separation medium has been developed with the aid of dimethyl sulfoxide (DMSO) as co-solvent. The best buffer conditions for primaquine, trihexyphenidyl (THP), sulconazole and cetirizine enantiomers were optimized as 20mM sodium phosphate buffer with DMSO/water (40/60, v/v) as solvent at a pH of 3.0, containing 10% (w/v) amylose. Partial-filling and semi-permanent coating techniques were used considering the influences of DMSO on UV detection. High chiral resolution for THP enantiomers was obtained showing good chiral separation capacity of this method. The method showed good linearity (R(2) > 0.998) over the concentration range of 0.50 and 2.00 mg L(-1) for all the enantiomers. The detection limits for the tested enantiomers were in the range from 0.05 to 0.12 mg L(-1). The linear calibration models were proven to be adequate for the experimental data by lack-of-fit test. The intra-assay precision, inter-day precision and accuracy were all evaluated to be acceptable. Separation and determination of THP enantiomers in rabbit blood were also carried out. PMID- 19135679 TI - High through-put aflatoxin determination in plant material by automated solid phase extraction on-line coupled to laser-induced fluorescence screening and determination by liquid chromatography-triple quadrupole mass spectrometry. AB - A screening test based on laser-induced fluorescence (LIF) and a method for individual identification - quantitation of aflatoxins (AFs) in olive leaves and drupes, based on chromatographic separation and triple-quad mass-spectrometry detection with electrospray ionization in positive mode, is here reported. The sensitivity and selectivity of both methods are enhanced by a preconcentration cleanup step developed by a Prospekt station. The analysis frequency is at least 3.5 samples/h. The screening test makes able to detect the target analytes at concentrations of 0.7microg/kg without "false negatives". The LC-MS/MS method provides limits of detection (LOD) and quantification (LOQ) ranging between 0.01 0.03 and 0.03-0.11microg/kg, respectively. The linear dynamic range is between LOQ-50microg/kg. The between-day precision, expressed as relative standard deviation, ranges between 0.97-2.86% and the within laboratory reproducibility, also expressed as RSD, between 1.63% and 4.84%. PMID- 19135680 TI - A combined QCM and XPS investigation of asphaltene adsorption on metal surfaces. AB - To investigate asphaltene-metal interactions, a combined quartz crystal microbalance (QCM) and X-ray photoelectron spectroscopy (XPS) study of asphaltene adsorption on a gold surface was conducted. Adsorption experiments were conducted at 25 degrees C with solutions of asphaltenes in toluene at concentrations ranging from 50 to 1500 ppm. QCM measurements yielded information on the kinetics of adsorption and further assessment of the data allowed the estimation of equilibrium adsorption levels. XPS analysis of adsorbed and bulk asphaltene demonstrated the presence of carboxylic, thiophenic, sulfide, pyridinic and pyrrolic type functional groups. The intensity of the main carbon (C-H) peak was related to surface coverage of adsorbed asphaltene as a function of asphaltene concentration by a simple mathematical model. The mass adsorption data from the QCM experiments also allowed estimation of the surface coverage, which was compared to those from XPS analyses. Surface coverage estimates as a function of asphaltene concentration could be described by a Langmuir (type-I) isotherm. The free energy of asphaltene adsorption was estimated to be -26.8+/-0.1 and -27.3+/ 0.1 kJ/mol from QCM and XPS data, respectively assuming asphaltene molar mass of 750 g/gmol. QCM and XPS data was also analyzed to estimate adsorbed layer thickness after accounting for surface coverage. The thickness of the adsorbed asphaltene estimated from both XPS and QCM data analyses ranged from 6-8 nm over the entire range of adsorption concentrations investigated. PMID- 19135681 TI - Acceleration effect of sulfate ion on the dissolution of amorphous silica. AB - The dissolution rate of amorphous silica is enhanced by sulfate ions. The zeta potential for silica particles in Na(2)SO(4) solution was lower than that in NaCl solution with the same ionic strength. These facts indicate that the specific adsorption of sulfate ions occurred by overcoming repulsion between negative charges of the SO(4)(2-) ion and SiO(-) on the surface of silica. The dissolution rate of amorphous silica may be accelerated by the specific adsorption of SO(4)(2 ) ions because of a decrease in the strength of the [triple bond]Si-O-Si[triple bond] bond in amorphous silica due to donation of electron density from the adsorbed SO(4)(2-) ions. PMID- 19135682 TI - Synthesis and characterization of arsenate-intercalated layered double hydroxides (LDHs): prospects for arsenic mineralization. AB - The arsenate-intercalated layered double hydroxide (LDH) of Mg and Al is synthesized by coprecipitation. The higher thermodynamic stability and the consequent lower solubility of the unitary arsenates preclude the formation of arsenate-intercalated LDHs of other metals directly from solution. However other M/Al-AsO(4) (M=Co, Ni, Zn) LDHs could be prepared by anion exchange, showing that arsenate intercalation proceeds topotactically. The intercalation of various species of As(V) into the interlayer of LDHs and the subsequent arsenate carrying capacity are dependent upon the pH of the solution. Upon thermal decomposition, the intercalated arsenate ion undergoes reductive deintercalation to give a mixture of As(III) and As(V) oxides. The product oxides revert back to the LDH upon soaking in water on account of the compositional and morphological metastability of the former. This is in contrast with the phosphate-intercalated LDHs, in which the reversibility is suppressed, consequent to the formation of stable metal phosphates. PMID- 19135683 TI - Effects of calcium ions on surface characteristics and adsorptive properties of hydrous manganese dioxide. AB - The effects of calcium ions (Ca(2+)) on the surface characteristics and adsorptive properties of hydrous manganese dioxide (deltaMnO(2)), taking arsenic and humic acid as model pollutants, were systematically investigated. The FTIR spectra and SEM/EDX analyses indicate coordination between Ca(2+) and the surface hydroxyls ([triple bond]Mn-OH) of deltaMnO(2), which leads to an increase of the surface zeta potential. Ca(2+) can facilitate deltaMnO(2) aggregation, as indicated by the particle size distribution (PSD) and on-line flocculating index (FI). The variation of deltaMnO(2) surface characteristics that is induced by Ca(2+) enhances the adsorption of humic acid (HA) and arsenic onto deltaMnO(2). The adsorption of HA is enhanced more significantly by Ca(2+) than that of arsenic, which is positively related to the coordination strength between Ca(2+) and individual pollutants. The SEM/EDX analysis shows the formation of Ca-HA complexes, which have a higher affinity for deltaMnO(2) than HA. The bridging effect of Ca(2+) plays a major role in enhancing HA adsorption onto deltaMnO(2). Due to different interactions between Ca(2+) and pollutants, the adsorption mechanism is pollutant-dependent in the presence of Ca(2+). PMID- 19135684 TI - Rural versus urban trauma: demographic influences on autopsy rates. AB - BACKGROUND: The autopsy has long been considered the gold standard for quality assurance review. Studies characterizing autopsies have been completed in large urban centers, but there is a paucity of research regarding autopsies at rural trauma centers. This is problematic considering that a majority of preventable trauma deaths occur in rural areas and death rates for unintentional injuries in rural populations are higher than urban populations. Rural trauma centers have differing characteristics warranting further research into the demographic differences between rural and urban trauma patients and the effects on autopsy rates. MATERIALS AND METHODS: This is a demographic study of a rural trauma center, University of Iowa Hospitals and Clinics (UIHC), with the goal of identifying characteristics of trauma patients on whom autopsy was performed. Four hundred ninety-six deaths were identified from the trauma registry between January 2002 and May 2007 (231 of which were autopsied) and demographic data (including age, race, length of hospital stay, etc.) regarding these patients was gathered into a database. Univariate and multivariate linear regression models were used to analyze differences between autopsied and non-autopsied trauma patients. Autopsy rate and basic demographics were also compared with 2 recent reports from urban trauma centers. RESULTS: Autopsied patients were younger than non-autopsied patients (mean age 45 y versus 71 y; P < 0.0001) and have a shorter median length of hospital stay (1 d versus 4 d; P < 0.0001). Autopsy rates for patients with blunt trauma were lower than rates for patients with penetrating or burn trauma (42% versus 67% and 56%; P = 0.004). If patients died while on a subspecialty service, they were less likely to have an autopsy. Compared with urban centers, this rural trauma center had lower autopsy rates, higher rates of blunt trauma, a higher mean age of deceased patients, and a lower percentage of males. CONCLUSIONS: UIHC, a rural trauma center, has a number of demographic characteristics that make it unique from urban trauma centers: an older population, lower percentage of male trauma patients, higher rates of blunt trauma, and lower rates of penetrating trauma. All of these factors influenced the lower rate of autopsies completed at rural trauma centers. Within a rural trauma center, those patients less likely to receive autopsy were older patients, those who died after 48 h in the hospital, and patients who suffered blunt injuries. The demographics of trauma patients most likely to receive an autopsy tend to correspond with those of an urban trauma population, thus providing a demographic explanation for the variation in autopsy rates among trauma systems. PMID- 19135685 TI - Sporadic primary hyperparathyroidism in young individuals: different disease and treatment? AB - BACKGROUND: Younger individuals with hyperparathyroidism may experience severe disease with a higher incidence of multigland disease (MGD) and operative failure, thereby requiring subtotal parathyroidectomy. This study examines the characteristics and surgical outcome of younger compared with older patients with sporadic primary hyperparathyroidism (SPHPT). METHODS: Prospectively collected data of 1101 patients with SPHPT who underwent parathyroidectomy at a single institution were retrospectively reviewed. Patients with multiple endocrine neoplasia (MEN), familial, secondary, or tertiary hyperparathyroidism, parathyroid carcinoma, rickets, or lithium induced disease were excluded. Patients were subdivided into two groups: (1) younger individuals < or = 40 y of age (n = 110) and (2) older individuals > 40 y of age (n = 991). Both age groups were compared for gender, clinical manifestations, pre- and postoperative laboratory values, MGD, operative success, and recurrent disease. RESULTS: There was greater male predominance in younger compared with older patients treated for SPHPT (41% versus 25%, P = 0.0004). Of the clinical manifestations of SPHPT, kidney stones were more common in younger compared with older individuals (45% versus 29%, P = 0.0006). Conversely, bone pain was more common in older compared with younger patients (32% versus 14%, P = 0.0002). There was no statistical difference in biochemical values, MGD, and outcome between both groups. CONCLUSIONS: Despite male predominance and few differences in symptoms, SPHPT is a similar disease entity in both younger and older individuals. Patients from both age groups can be similarly treated for SPHPT with a high rate of operative success. Routine BNE and subtotal parathyroidectomy is not necessary in younger individuals. PMID- 19135686 TI - Frequency and clinical patterns of multiple sclerosis in Arab countries: a systematic review. AB - The susceptibility of various populations to multiple sclerosis (MS) and the clinical patterns of the disease are thought to be different. Nineteen articles related to incidence, prevalence and clinical patterns of MS in Arab populations were identified by keyword searching of Medline and Embase, and review of references in all relevant papers. Data were only available for the Kuwaiti, Jordanian, Libyan, Saudi, Iraqi, Palestinian (including Arabs living in Israel), and Omani populations. The publications ranged from 1975 to 2007. In Israel the incidence of MS was 0.7 per 100,000 per year in Arabs born and living in Greater Jerusalem. In Kuwait, the incidence of MS was 2.08 per 100,000. Prevalence varied from 4 to 42 per 100,000 population. The clinical pattern of MS was generally similar to that in western countries. However, one study from Oman found a high rate of optic-spinal disease (affecting one third of patients) and a low rate of oligoclonal bands (OGBs) (only one third of patients); this pattern resembles that of MS described in Asian countries. In conclusion, the prevalence of MS among Arabs has a wide reported range. The clinical pattern is generally similar to "Western type" MS but apparent differences in optic-spinal disease and OGBs positivity need further evaluation. There is significant opportunity for further evaluation of MS in Arabs, especially in unstudied areas, including the populous countries of Egypt, Algeria, Syria, and Morocco. Studies of Arab-Americans and Arab immigrants in Europe could help in defining the effect of immigration on MS. Such studies are likely to enhance our knowledge of the environmental, genetic and clinical variation of MS in Arabs. PMID- 19135687 TI - Impact of a coastal dump in a tropical lagoon on trace metal concentrations in surrounding marine biota: a case study from Saipan, Commonwealth of the Northern Mariana Islands (CNMI). PMID- 19135688 TI - Leatherback turtles: the menace of plastic. AB - The leatherback, Dermochelyscoriacea, is a large sea turtle that feeds primarily on jellyfish. Floating plastic garbage could be mistaken for such prey. Autopsy records of 408 leatherback turtles, spanning 123 years (1885-2007), were studied for the presence or absence of plastic in the GI tract. Plastic was reported in 34% of these cases. If only cases from our first report (1968) of plastic were considered, the figure was 37%. Blockage of the gut by plastic was mentioned in some accounts. These findings are discussed in the context of removal of top predators from poorly understood food chains. PMID- 19135689 TI - The use of statistical methods to separate natural background and anthropogenic concentrations of trace elements in radio-chronologically selected surface sediments of the Venice Lagoon. AB - A new method has been developed that predicts the natural background concentrations of microelements of interest in sediments using the concentrations of various components present in the sample. This is done by developing multiple linear regressions using a dataset which has been selected, using radiochronological dating and statistical methods, to reflect the concentration distributions of natural levels of a large number of sediment constituents. This method is demonstrated in Venice Lagoon, using data generated in a lagoon-wide sampling effort. A quantitative assessment is carried out of the variability of natural background levels at the lagoon scale for the main inorganic microelements, and distribution maps of the background and anthropogenic concentrations of a selected metal (Ni) in sediments at the lagoon scale are presented. With sufficient data, this method can be applied in other areas, allowing for a site-specific separation of natural background and anthropogenic levels of microconstituents in sediments, providing insights into regional geochemistry and aiding in the risk-based management of coastal sediments. PMID- 19135690 TI - Cassane diterpenoids from the stem of Caesalpinia pulcherrima. AB - Cassane diterpenoids: pulcherrin A, pulcherrin B, pulcherrin C, neocaesalpin P, neocaesalpin Q and neocaesalpin R, together with eight known compounds: isovouacapenol C, 6beta-cinnamoyl-7beta-hydroxy-vouacapen-5alpha-ol, pulcherrimin E, pulcherrimin C, alpha-cadinol, 7-hydroxycadalene, teucladiol and bonducellin were isolated from the stem of Caesalpinia pulcherrima. The chemical structures were elucidated by analysis of their spectroscopic data. PMID- 19135691 TI - Measuring the health of urban populations: what progress have we made? AB - This paper considers the progress that has been made over the past 15 years in establishing an information base for the measurement of health variations, and for the assessment of the health needs of vulnerable populations living in urban areas. Four small areas, two in Coventry and two in Leicester, were explored. An extensive exercise in data trawling was undertaken and the available data were examined. On the basis of this exercise, the findings and recommendations of a paper published 15 years ago were revisited to assess the extent of the progress that had been made in the development of appropriate data sources. It was found that considerable progress had been made in provision of data relating to health status and health determinants, but that knowledge of small area populations had failed to keep up with the changing demographic structures of recent years. As a consequence, the possibilities for the exploitation and interpretation of health status and health determinant data are limited. An approach to data collation which combines available quantitative and qualitative data sources is currently being trialled in one of the four areas studied. It is argued that this provides the approach that will be most effective for the foreseeable future. PMID- 19135692 TI - 'Partners in clime': sustainable development and climate change--what can the National Health Service do? AB - Climate change is arguably the biggest threat to health in the medium and long term. Necessary responses to this threat include adaptation, i.e. preparing to respond to the consequences of climate change, of which there are many in respect of health; and mitigation, i.e. reducing the activities that lead to climate change and, in particular, reducing the levels of greenhouse gas emissions, the most significant of which is carbon. PMID- 19135693 TI - Beyond the 'nanny state': stewardship and public health. AB - BACKGROUND: Some public health measures restrict personal freedom more than others, and deciding what type of measure will be appropriate and effective has long been a problem for policy makers. Existing bioethical frameworks are often not well suited to address the problems of public health. METHODS: The Nuffield Council on Bioethics set up an expert working party to examine the ethical issues surrounding public health in January 2006. Following evidence gathering and a public consultation exercise, the Council published its conclusions and recommendations in the report 'Public health: ethical issues' in November 2007. RESULTS: A spectrum of views exists on the relationship between the state's authority and the individual. The Council set out a proposal to capture the best of the libertarian and paternalistic approaches, in what it calls the 'stewardship model'. This model suggests guiding principles for making decisions about public health policies, and highlights some key principles including Mill's harm principle, caring for the vulnerable, autonomy and consent. An 'intervention ladder' is also proposed, which provides a way of thinking about the acceptability of different public health measures. The report then applies these principles to a number of case studies: infectious diseases, obesity, alcohol and tobacco, and fluoridation of water supplies. CONCLUSIONS: The idea of a 'nanny state' is often rejected, but the state has a duty to look after the health of everyone, and sometimes that means guiding or restricting people's choices. On the other hand, the state must consider a number of principles when designing public health programmes, and justification is required if any of these principles are to be infringed. PMID- 19135694 TI - Transport and health--a five-country perspective. AB - Devolution in the UK has, inadvertently, provided a natural experiment for comparing the impact of different approaches to policy and practice on populations that are broadly similar by most criteria. Realizing the potential gold mine that this presents for comparing policy and practice in the field of public health, the UK Public Health Association has established a Devolution Strategic Interest Group to explore and capitalize on this. From the start, the Group has included the Republic of Ireland in its purview to add an extra dimension of diversity and innovation. Having previously looked at the organization of health care and public health, and at policy and practice on food, nutrition and obesity, the Group opted to look at transport and health this year. Here, the links between separate relevant policy streams are less obvious and clearly less developed than those for food, nutrition and obesity. Also, the perceived need to raise the issue up the public health agenda provides a new challenge to campaigning action for the Group. PMID- 19135695 TI - West Nile virus infection modulates human brain microvascular endothelial cells tight junction proteins and cell adhesion molecules: Transmigration across the in vitro blood-brain barrier. AB - Neurological complications such as inflammation, failure of the blood-brain barrier (BBB), and neuronal death contribute to the mortality and morbidity associated with WNV-induced meningitis. Compromised BBB indicates the ability of the virus to gain entry into the CNS via the BBB, however, the underlying mechanisms, and the specific cell types associated with WNV-CNS trafficking are not well understood. Brain microvascular endothelial cells, the main component of the BBB, represent a barrier to virus dissemination into the CNS and could play key role in WNV spread via hematogenous route. To investigate WNV entry into the CNS, we infected primary human brain microvascular endothelial (HBMVE) cells with the neurovirulent strain of WNV (NY99) and examined WNV replication kinetics together with the changes in the expressions of key tight junction proteins (TJP) and cell adhesion molecules (CAM). WNV infection of HBMVE cells was productive as analyzed by plaque assay and qRT-PCR, and did not induce cytopathic effect. Increased mRNA and protein expressions of TJP (claudin-1) and CAM (vascular cell adhesion molecule and E-selectin) were observed at days 2 and 3 after infection, respectively, which coincided with the peak in WNV replication. Further, using an in vitro BBB model comprised of HBMVE cells, we demonstrate that cell-free WNV can cross the BBB, without compromising the BBB integrity. These data suggest that infection of HBMVE cells can facilitate entry of cell-free virus into the CNS without disturbing the BBB, and increased CAM may assist in the trafficking of WNV-infected immune cells into the CNS, via 'Trojan horse' mechanism, thereby contributing to WNV dissemination in the CNS and associated pathology. PMID- 19135696 TI - Effect of ozone exposure on the oxidation of trace organic contaminants in wastewater. AB - Three tertiary-treated wastewater effluents were evaluated to determine the impact of wastewater quality (i.e. effluent organic matter (EfOM), nitrite, and alkalinity) on ozone (O(3)) decomposition and subsequent removal of 31 organic contaminants including endocrine disrupting compounds, pharmaceuticals, and personal care products. The O(3) dose was normalized based upon total organic carbon (TOC) and nitrite to allow comparison between the different wastewaters with respect to O(3) decomposition. EfOM with higher molecular weight components underwent greater transformation, which corresponded to increased O(3) decomposition when compared on a TOC basis. Hydroxyl radical (()OH) exposure, measured by parachlorobenzoic acid (pCBA), showed that limited ()OH was available for contaminant destruction during the initial stage of O(3) decomposition (t<30s) due to the effect of the scavenging by the water quality. Advanced oxidation using O(3) and hydrogen peroxide did not increase the net production of ()OH compared to O(3) under the conditions studied. EfOM reactivity impacted the removal of trace contaminants when evaluated based on the O(3):TOC ratio. Trace contaminants with second order reaction rate constants with O(3)(k(O)(3))>10(5)M( 1)s(-1) and ()OH (k(OH))>10(9)M(-1)s(-1), including carbamazepine, diclofenac, naproxen, sulfamethoxazole, and triclosan, were >95% removed independent of water quality when the O(3) exposure (integralO(3)t) was measurable (0-0.8mgmin/L). O(3) exposure would be a conservative surrogate to assess the removal of trace contaminants that are fast-reacting with O(3). Removal of contaminants with k(O)(3) < 10M(-1)S(-1) , and k(OH)>10(9)M(-1)s(-1), including atrazine, iopromide, diazepam, and ibuprofen, varied when O(3) exposure could not be measured, and appeared to be dependent upon the compound specific k(OH). Atrazine, diazepam, ibuprofen and iopromide provided excellent linear correlation with pCBA (R(2)>0.86) making them good indicators of ()OH availability. PMID- 19135697 TI - Whole lake selective withdrawal experiment to control harmful cyanobacteria in an urban impoundment. AB - Different environmental conditions support optimal growth by Aphanizomenon and Microcystis in Ford Lake, Michigan, USA, based on weekly species biovolume and water chemistry measurements from June through October 2005-2007. Experimental withdrawal of hypolimnetic water through the outlet dam was conducted in 2006, with 2005 and 2007 acting as control years, to test theory regarding management of nuisance and toxic cyanobacteria. The dynamics of Aphanizomenon and Microcystis blooms in Ford Lake appear to be driven largely by NO3(-) concentrations, with higher levels shifting the advantage to Microcystis (P<0.0001). Aphanizomenon was most successful with a mean TN:TP ratio (mol:mol) of 48.3:1, whereas Microcystis thrived with a mean ratio of 70.1:1. Withdrawal of hypolimnetic water successfully destabilized the water column and led to higher levels of NO3(-) and the near elimination of the Aphanizomenon bloom in 2006 (P<0.0001). Selective withdrawal did not reduce Microcystis biovolume or microcystin toxicity. Microcystis biovolume and NO3(-) levels were positively correlated with microcystin toxin (P=0.01) and jointly accounted for 30.5% of the variability in the data. Selective withdrawal may be a viable management option for improving water quality under certain circumstances. To fully address the problem of nuisance and toxic algal blooms in Ford Lake, however, an integrated approach is required that targets cyanobacteria biovolume dynamics as well as conditions suited for toxin production. PMID- 19135698 TI - CFD simulation of mechanical draft tube mixing in anaerobic digester tanks. AB - Computational Fluid Dynamics (CFD) was used to simulate the mixing characteristics of four different circular anaerobic digester tanks (diameters of 13.7, 21.3, 30.5, and 33.5m) equipped with single and multiple draft impeller tube mixers. Rates of mixing of step and slug injection of tracers were calculated from which digester volume turnover time (DVTT), mixture diffusion time (MDT), and hydraulic retention time (HRT) could be calculated. Washout characteristics were compared to analytic formulae to estimate any presence of partial mixing, dead volume, short-circuiting, or piston flow. CFD satisfactorily predicted performance of both model and full-scale circular tank configurations. PMID- 19135699 TI - Disturbances in energy metabolism of Daphnia magna after exposure to tebuconazole. AB - This study was conducted to investigate the change of some biochemical parameters in the aquatic invertebrate Daphnia magna following exposure to the fungicide tebuconazole and to determine the most sensitive biomarker among the ones tested in this species. Four biochemical biomarkers (protein, glycogen, lipids and caloric content) were correlated with feeding behaviour studies of D. magna after fungicide exposure. Juveniles of D. magna were exposed to four sublethal concentrations of tebuconazole (0.41, 0.52, 0.71 and 1.14 mgL(-1)) for 5d. Daphnid samples were taken from each test and control group at 24, 48, 72, 96 and 120 h after the start of the experiment. Tebuconazole EC(50) values were calculated on D. magna in our laboratory as 56.83 and 40.10 mgL(-1) at 24 and 48 h, respectively. Results showed that daphnid energy content decreased as tebuconazole concentration increased, especially after 96-120 h of exposure to 0.52 mgL(-1) and higher fungicide concentrations. The data suggest that tebuconazole is moderately toxic to D. magna but also that it seriously impairs the metabolic functions, resulting in alterations in biochemical constituents. In the D. magna feeding study, algae feeding rates were inhibited after fungicide exposure. Such findings indicate the importance of feeding studies in laboratory toxicity test as well as their relationship with others studies. The results emphasize the importance of considering different kind of biomarkers to identify and evaluate the biological effect of a fungicide in the aquatic environment. Although the biochemical biomarkers used resulted good indicators of tebuconazole toxicity, feeding rates in D. magna decreased after only 5h exposure to the fungicide resulting in the most sensitive parameter of daphnid fungicide exposure. PMID- 19135700 TI - Mineral induced mechanochemical degradation: the imazaquin case. AB - The potential role of mechanochemical processes in enhancing degradation of imazaquin by soil components is demonstrated. The investigated components include montmorillonite saturated with Na(+), Ca(2+), Cu(2+)and Al(3+), Agsorb (a commercial clay mix), birnessite and hematite. The mechanical force applied was manual grinding of mixtures of imazaquin and the minerals, using mortar and pestle. The degradation rates of imazaquin in these mixtures were examined as a function of the following parameters: time of grinding, herbicide load (3.9, 8.9, 16.7 and 26.6 mg imazaquin per g mineral), temperature (10, 25, 40 and 70 degrees C), acidic/basic conditions, and dry or wet grinding. Dry grinding of imazaquin for 5 min with Al-montmorillonite or with hematite resulted in 56% and 71% degradation of the imazaquin, respectively. Wet grinding slightly reduced the degradation rate with hematite and entirely cancelled the enhancing effect of grinding with Al-montmorillonite. Wet grinding in the presence of the transition metals: Ni(2+), Cu(2+), Fe(3+) added as chlorides was carried out. Addition of Cu(2+) to Na-montmorillonite loaded with imazaquin was the most effective treatment in degrading imazaquin (more than 90% of the imazaquin degraded after 5 min of grinding). In this treatment, Cu-montmorillonite formation during the grinding process was confirmed by XRD and accordingly, grinding with Cu montmorillonite gave similar degradation values. LC-MS analysis revealed that the mechanochemical transformation of imazaquin resulted in the formation of a dimer and several breakdown products. The reported results demonstrate once again that mechanochemical procedures offer a remediation avenue applicable to soils polluted with organic contaminants. PMID- 19135701 TI - Effect of perfluorooctane sulfonate on toxicity and cell uptake of other compounds with different hydrophobicity in green alga. AB - Perfluorooctane sulfonate (PFOS) was evaluated alone and in binary mixtures with pentachlorophenol, atrazine and diuron, respectively to investigate the effects of interactions between PFOS and other compounds on the growth rate in Scenedesmus obliquus. Single application of PFOS showed no inhibition on the growth of S. obliquus below 40 mg L(-1), whereas PFOS acting with pentachlorophenol resulted in higher algal growth inhibition in comparison with pentachlorophenol alone. A maximum increase of 45% in the growth inhibition was observed at a pentachlorophenol concentration of 2.56 mg L(-1) together with a PFOS concentration of 40 mg L(-1). On the contrary, the algal growth inhibition of atrazine and diuron was depressed by PFOS. Furthermore, cell uptake was examined to gain some insights into the mechanisms of the effects of PFOS on the toxicity of the other compounds. Cell uptake of pentachlorophenol increased while that of atrazine and diuron was reduced in cells that have been exposed to PFOS. The effects of PFOS on the toxicity of pentachlorophenol, atrazine and diuron were possibly related to the influence of PFOS on the cell uptake of these hydrophobic compounds. Results suggested that PFOS influenced the cell uptake and toxicity of structurally different compounds in dissimilar manners and potentially increased the accessibility and toxicity of more hydrophobic compounds to algal cells. PMID- 19135702 TI - The impacts of cypermethrin pesticide application on the non-target microbial community of the pepper plant phyllosphere. AB - Although pesticides have been extensively used for controlling insects and disease pathogens of plants, little is known regarding the impacts of applying these pesticides on the microbial community in the plant phyllosphere. Here, we report the effects of cypermethrin pesticide application upon the microbial community of the pepper plant phyllosphere. Assessments were made using culture independent techniques including phospholipid fatty acid analysis (PLFA) and 16S rRNA gene directed Polymerase Chain Reaction with Denaturing Gradient Gel Electrophoresis (PCR-DGGE). During the 21 day greenhouse study, PLFA results indicated that both total and bacterial biomass increased after application of the pesticide. PLFA profiles also indicated that Gram-negative bacteria became predominant. DGGE analysis confirmed a significant change in bacterial community structure within the phyllosphere following the pesticide application where different dendrogram clusters were observed between control and treated samples. Phylogenetic analysis also suggested a change in bacterial phyla following treatment, where bands sequenced within control cultures were predominantly of the Firmicutes phylum, but those bands sequenced in the treated samples were predominantly members of the Bacteroidetes and gamma-Proteobacteria phyla. In conclusion, this study revealed an increase in bacterial abundance and a shift in community composition within the pepper plant phyllosphere following the pesticide application, and highlighted the effective use of PLFA and PCR-DGGE for studying the effect of pesticides upon indigenous phyllosphere microbes. PMID- 19135703 TI - The checkered puffer (Spheroides testudineus) and its helminths as bioindicators of chemical pollution in Yucatan coastal lagoons. AB - The suitability of using helminth communities as bioindicators of environmental quality of the Yucatan coastal lagoons status was tested on the checkered puffer (Spheroides testudineus) in four coastal lagoons along the Yucatan coast. The concentration of chemical pollutants in sediments, water quality parameters, helminth infracommunity characteristics, as well as fish physiological biomarkers, including EROD (7-ethoxyresorufin-O-deethylase) and catalase activities, were measured. Results from sediment analyses demonstrated the presence of hydrocarbons, organochlorine pesticides and polychlorinated biphenyls at varying concentrations, some of which exceeded the Probability Effect Level (PEL). Significant negative associations among organochlorine pesticides, infracommunity characteristics and fish physiological responses were observed in most of the lagoons. Results suggest that EROD activity and parasite infracommunity characteristics could be useful tools to evaluate the effects of chemical pollutants on the fish host and in the environment. Importantly, certain parasites appear to influence biomarker measurements, indicating that parasites should be considered in ecotoxicological studies. PMID- 19135704 TI - The coagulopathy of cirrhosis assessed by thromboelastometry and its correlation with conventional coagulation parameters. AB - BACKGROUND: Thromboelastometry allows continuous registration of the blood viscoelastic changes upon activation by cephaline or tissue-factor plus calcium chloride. The technique is used as a near-patient-testing device to guide transfusion in cardiac surgery or liver transplantation and less to investigate hemostasis in acquired or congenital coagulopathies. AIMS: (i) Review of the coagulopathy associated with cirrhosis and (ii) report on its investigation by thromboelastometry in comparison with conventional coagulation parameters. METHODS: We investigated citrated blood samples from 51 adult cirrhotics for the following thromboelastometry parameters: coagulation-time (CT), clot-formation time (CFT), maximum-clot-firmness (MCF). RESULTS: Relatively few patients [14/51(27%)] were identified as abnormal by CT; in contrast, a greater proportion were identified by the CFT [41/51(80%)] or MCF [39/51(76%)]. CFT and MCF were correlated with the platelet-count, antithrombin and fibrinogen. Prothrombin time (PT) was correlated with CFT and MCF. None of the coagulation parameters were correlated with CT. The correlation of the Child-Pugh-score (taken as index of severity) versus MCF or PT was -0.457(p < 0.001) or 0.484(p < 0.001), suggesting MCF as a suitable prognostic index. CFT and MCF, but not CT obtained ROC curves that were useful to distinguish patients from healthy individuals. CONCLUSIONS: Thromboelastometry, currently used to assist liver transplantation is also suitable for investigating stable cirrhosis. CFT and MCF are the most interesting parameters to be considered for future clinical studies needed to assess their value as measures of bleeding-risk and prognosis in this category of patients. PMID- 19135706 TI - Characterization of a homozygous Gly11Val mutation in the Gla domain of coagulation factor X. AB - Factor (F) X deficiency is a rare inherited autosomal recessive trait. We report on a patient affected by a severe bleeding diathesis. Mutations were sought by F10 sequence analysis. The consequences of the mutation were characterized by measuring thrombin and FXa formation after triggering the clotting cascade with activated partial thromboplastin time (aPTT) reagent or with phospholipid vesicles plus either tissue factor (TF) or FIXabeta, or with the FX activator from Russell's viper venom (RVV-X). The patient was found to be homozygous for a novel FX p.G51V mutation (G11V of the mature protein) within the omega-loop of the gamma-carboxyglutamic-rich domain. FX activity was markedly reduced (FX:C <1%) in prothrombin time and aPTT assays, and was 15% of normal in the RVV-X assay. The antigen level (FX:Ag) was 75%. TF, alone or in combination with recombinant FVIIa, failed to trigger detectable FXa or thrombin activity in the patient's plasma. FIXabeta also failed to trigger measurable FXa or thrombin production, but activation with RVV-X was only 4-fold less effective in the patient's plasma than in normal plasma. Supplementation with normal FX suggested that FX(G11V) and/or FXa(G11V) might slow the clotting cascade by competition. Overall, the patient's phenotype appears to be due to a very low rate of FX(G11V) activation by TF/FVIIa and FVIIIa/FIXa complexes rather than to FXa(G11V) activity within prothrombinase. PMID- 19135705 TI - The effect of antiplatelet drugs clopidogrel and aspirin is less immediately after stent implantation. AB - INTRODUCTION: A decreased effect of clopidogrel or aspirin on platelets corresponds to an increased risk of major adverse coronary events. The aim was to investigate if the inhibition of platelet function by clopidogrel and aspirin is equal at two different time points: immediately after percutaneous coronary intervention (PCI) and one day thereafter. MATERIALS AND METHODS: Platelet function was assessed by the Vasodilator Stimulated Phosphoprotein (VASP) phosphorylation assay, Impedance Aggregometry, Platelet Function Analyzer and the Cone and Platelet Analyzer in 30 patients on chronic treatment with clopidogrel and aspirin. RESULTS: Inhibition of platelets by clopidogrel and aspirin was less post PCI than one day after PCI as measured with the VASP assay and aggregometry: the platelet reactivity index, the adenosine diphosphate/prostaglandin and the arachidonic acid -induced platelet aggregation were 23% (p=0.009), 75% (p=0.001) and 127% (p<0.001) higher post PCI than one day after PCI, respectively. The collagen/adenosine diphosphate closure time was 30% higher after PCI compared to one day thereafter (p=0.047), which could in part be due to a two-fold increase in von Willebrand factor-ristocetin cofactor activity one day after PCI (p=0.001). CONCLUSION: Inhibition of platelets by clopidogrel and aspirin was less immediately post PCI as compared to one day thereafter. This indicates that the time point of platelet function testing is important for the determination of cut-off points and the definition of nonresponsiveness to antiplatelet drugs. PMID- 19135707 TI - A case of vesicocutaneous fistula to the thigh. AB - We report a case of vesicocutaneous fistula to the thigh that developed 7 years after an injury to the pelvis. A 34-year-old male patient was referred to our hospital because of a water-soluble yellow exudate from his left thigh. A urinary fistula was suggested by the results of an intravenous indigo carmine injection. Intravenous pyelography and computed tomography revealed the presence of a bladder deformation and a fistula originating at the ventral bladder wall and leading down to the left thigh. The patient underwent fistulectomy and partial cystectomy. To the best of our knowledge, no previous reports of vesicocutaneous fistula to the thigh have been published. PMID- 19135708 TI - Adjuvant gemcitabine plus docetaxel for completely resected stages I-IV high grade uterine leiomyosarcoma: Results of a prospective study. AB - OBJECTIVE: Patients with completely resected stages I-IV high grade uterine leiomyosarcoma are at high risk for recurrence. No adjuvant treatment has been shown to improve survival, although prospective data are limited. We sought to determine whether adjuvant gemcitabine-docetaxel would yield a 2-year progression free survival of at least 50% in this leiomyosarcoma population. METHODS: Eligible patients were treated with gemcitabine 900 mg/m(2) over 90 min days 1 and 8 plus docetaxel 75 mg/m(2) day 8, every 3 weeks for 4 cycles. CT imaging was performed at baseline, after cycle 4, and every 3 months. Progression was defined as evidence of new disease on CT. RESULTS: Twenty-five patients (median age 49; range, 37-73) enrolled; 23 were evaluable (1-never treated, 1-ineligible). With median follow-up of 49 months for all patients, 10 (45%) of the 23 evaluable patients remained progression free at 2 years, with a median progression-free survival of 13 months. The median overall survival is not yet reached. Among the 18 patients with stages I or II uterine leiomyosarcoma, 59% remain progression free at 2 years, with a median progression-free survival of 39 months. Median overall survival for stages I and II patients is not yet reached with median follow-up duration of 49 months. Sites of first recurrence were: lung only - 3/23 (13%); pelvis only - 5/23 (22%); both - 5 (22%). CONCLUSIONS: Post-resection gemcitabine-docetaxel for stages I-IV high-grade uterine leiomyosarcoma yields 2 year progression-free survival rates that appear superior to historical rates. Gemcitabine-docetaxel merits further study as part of an adjuvant strategy for patients with completely resected, early-stage uterine leiomyosarcoma. PMID- 19135710 TI - Multicellular spheroids in ovarian cancer metastases: Biology and pathology. AB - Epithelial ovarian cancer (EOC) has a relatively high mortality rate ( approximately 55%). One of the presiding causes is that the current chemotherapeutic regimes are unable to achieve sustained remission, despite frequently producing a positive response at first treatment. One of the reasons that EOC is difficult to treat is that the mechanism of dissemination is unusual. EOC dissemination characteristically involves local invasion of pelvic and abdominal organs. Unlike many epithelial cancers, initial dissemination rarely requires the vasculature, although the vasculature is often implicated in the advanced stages of disease. Recently, it has become apparent that aggregates of malignant cells (spheroids) contained within malignant ascites represent a significant impediment to efficacious treatment of late stage EOC. In vivo, spheroids are present in the malignant ascites of EOC patients, while in vitro cultured spheroids are capable of tumorgenesis in vivo and display a reduced response to chemotherapeutic drugs when compared to monolayers. A major problem associated with the current generation of chemotherapy agents is that they do not address the anchorage- and vascular-independent growth conditions associated with a 3-dimensional structure that has formed and/or grown in suspension. Thus, spheroid formation may represent a key component of platinum/taxane-sensitive recurrence. If this is correct, a better understanding of spheroid biology may contribute to the identification of new treatment opportunities for the sustained treatment of metastatic EOC. This review article outlines the key biological features of spheroids, specifically discussing their role in EOC dissemination and chemo-response as well as providing insights into spheroid functionality. PMID- 19135709 TI - Evaluation of an intraperitoneal chemotherapy program implemented at the Princess Margaret Hospital for patients with epithelial ovarian carcinoma. AB - OBJECTIVE: A prospective evaluation of an ambulatory intraperitoneal (IP) /intravenous (IV) chemotherapy regimen for women with epithelial ovarian carcinoma (EOC). METHODS: Cisplatin 100 mg/m(2) (option for 75 mg/m(2)) IP combined with paclitaxel 175 mg/m(2) IV (3 h infusion) administered every 21 days was adopted by our institution as a single day, outpatient regimen for women with stage III EOC who had undergone optimal cytoreductive (or=upper quartile). Immunoblot expression of MASPIN, THBS-1, bFGF, VEGF, VEGFR-1 and p53 status (mutation and overexpression) was previously determined. RESULTS: Of 106 evaluable cases, 25% exhibited high CD31-MVD (>24.25 vessels/high power field [HPF]) or high CD105-MVD (>19.25 vessels/HPF). After adjusting for age and stratifying by GOG performance status, stage, cell type, grade, debulking status and treatment regimen, high versus low CD105-MVD was associated with increased risk of disease progression (hazard ratio [HR]=1.873; 95% confidence interval [CI]: 1.102-3.184; p=0.020), but not death (HR=1.125; 95% CI: 0.654-1.935; p=0.670) whereas CD31-MVD was not associated with risk of disease progression (HR=1.578; 95% CI=0.918-2.711; p=0.099) or death (HR=1.678; 95% CI=0.957-2.943; p=0.071). CD31-MVD was correlated with CD105-MVD (p=0.001) and MASPIN (p=0.016). Neither CD31-MVD nor CD105-MVD was associated with p53 status, THBS-1, bFGF, VEGF or VEGFR-1. CONCLUSIONS: High MVD assessed using CD105, a marker of proliferating endothelial cells and neoangiogenesis, but not CD31 a pan-endothelial marker, appeared to be an independent prognostic factor for worse progression-free survival in women with advanced EOC after adjusting for prognostic clinical covariates. PMID- 19135713 TI - Clonal mast cell disorders in patients with systemic reactions to Hymenoptera stings and increased serum tryptase levels. AB - BACKGROUND: Anaphylaxis after Hymenoptera stings has been reported in subjects with mastocytosis, but few data exist regarding disease prevalence in populations allergic to these insects. OBJECTIVE: The incidence of clonal mast cell (MC) disorders in subjects with both systemic reactions to Hymenoptera stings and increased serum baseline tryptase (sBT) levels was assessed by using bone marrow (BM) aspirates and biopsy specimens. METHODS: Subjects with a history of a systemic reaction caused by a Hymenoptera sting underwent the standard diagnostic work-up for Hymenoptera allergy, and sBT levels were measured. Subjects with an increased sBT level had BM evaluation that included histology/cytology, flow cytometry, and detection of KIT mutations. RESULTS: Forty-four (11.6%) of 379 subjects with systemic reactions had increased sBT levels (>11.4 ng/mL), and 31 (70.5%) of these had a history of anaphylaxis. Thirty-four subjects with increased sBT levels underwent a BM analysis. Histology detected diagnostic or subdiagnostic MC infiltrates in 22 (65%) of 34 patients. Abnormal MCs were identified by means of flow cytometry and cytology in 26 (78.8%) of 33 and 20 (58.8%) of 34 subjects, respectively. A KIT mutation was detected in 17 (54.8%) of 31 subjects. The diagnosis was indolent systemic mastocytosis in 21 (61.7%) of 34 subjects and monoclonal MC activation syndrome in 9 (26.5%) of 34 subjects. All subjects with anaphylaxis had one of those 2 disorders. CONCLUSION: The concomitant presence of systemic reactions (especially anaphylaxis) after Hymenoptera stings and increased sBT levels strongly suggests that a BM examination is indicated for the diagnosis of clonal MC disease. PMID- 19135714 TI - Birth of domestic cat kittens of predetermined sex after transfer of embryos produced by in vitro fertilization of oocytes with flow-sorted sperm. AB - Our goals were to: (1) determine if domestic cat sperm could be sorted to high purity by flow cytometry after overnight shipment of cooled samples; (2) evaluate the efficiency with which sorted sperm could be used to generate cat embryos in vitro; and (3) determine if live kittens of predetermined sex could be produced after transfer of embryos derived by IVF using sorted sperm. Semen samples (n=5) from one male were extended in electrolyte-free solution and shipped overnight at 4 degrees C to the sorting facility. Samples were adjusted to 75x10(6)sperm/mL and stained with Hoechst 33342. After 1h at 34.5 degrees C, samples were adjusted to 50x10(6)sperm/mL with 4% egg yolk TALP+0.002% food dye and sorted by high speed flow cytometry. Later resort analysis confirmed purities of 94% and 83% for X- and Y-chromosome bearing sperm, respectively. Sorted sperm were centrifuged, re-suspended in TEST yolk buffer and shipped overnight to the IVF laboratory. After IVF of in vivo matured oocytes with X-chromosome bearing sperm, cleavage frequency was 62% (54/87). After IVF of IVM oocytes with control, X- or Y chromosome bearing sperm, the incidence of cleavage was 42% (48/115), 33% (40/120), and 35% (52/150), respectively, and blastocyst development was 53% (21/40), 50% (11/22), and 55% (23/42), respectively (P>0.05). On Day 2, 45 embryos produced by IVF of in vivo matured oocytes with X-chromosome bearing sperm were transferred to the oviduct of four Day 1 recipients, three of which subsequently delivered litters of one, four, and seven female kittens, respectively. In conclusion, we confirmed that sperm sorting technology can be applied to domestic cats and established that kittens of predetermined sex can be produced. PMID- 19135715 TI - Effect of water-bath post-polymerization on the mechanical properties, degree of conversion, and leaching of residual compounds of hard chairside reline resins. AB - OBJECTIVES: This study evaluated the effect of water-bath post-polymerization at 55 degrees C for 10 min (WB) on the content and leaching of residual compounds, degree of conversion, flexural strength, and hardness of hard chairside reline resins (Kooliner: K, New Truliner: N, Ufi Gel hard: U, and Tokuso Rebase Fast: T). METHODS: Leaching experiments were made by storing specimens (n=48) in artificial saliva at 37+/-1 degrees C and analyzing residual monomers and plasticizer by HPLC. Analysis of residual monomer and plasticizer content (n=48) was also made by HPLC. Degree of conversion (n=40) was analyzed by using FT-Raman spectroscopy. A 3-point loading test was used to evaluate the flexural strength of the specimens (n=80). One fragment of each flexural test specimen was then submitted to Vickers microhardness test. RESULTS: WB produced a significant decrease (p<0.050) in the amount of residual compounds eluted from the materials within the first hour of immersion. With the exception of material U, WB decreased the duration of release of the residual compounds evaluated. All materials evaluated exhibited significantly (p<0.050) lower values of residual monomer and plasticizer (material N) after WB compared with the control groups. WB increased the degree of conversion of K and T resins and the hardness of N, K, and T resins (p<0.050). Only material K showed an increase in flexural strength after WB (p<0.050). SIGNIFICANCE: Immersion of relined dentures in water at 55 degrees C for 10 min can be used to reduce the amounts and duration of release of residual compounds and improve mechanical properties of some of the materials evaluated. PMID- 19135716 TI - Augmentation of skeletal tissue formation in impaction bone grafting using vaterite microsphere biocomposites. AB - The development of particulate bone void fillers with added biological function to augment skeletal tissue formation will lead to improved efficacy in bone replacement surgery. We demonstrate the potential for vaterite microsphere biocomposites to augment bone matrix formation within an in vivo model for impaction bone grafting seeded with human bone marrow stromal cells. In vitro tests demonstrate the significance of vaterite microspheres in the activation and promotion of 3D skeletal tissue formation. Further in vitro experiments using functionalized microspheres with surface integrated RGD peptide activate co cultured skeletal populations in pellets and promote secretion of extracellular matrix collagens and human osteocalcin. Specific temporal release of entrapped RNase A was successfully demonstrated using these specialized microspheres with integrated magnetic beads, which physically disrupted the inorganic macrostructure. These studies demonstrate that bio-inspired calcium carbonate microspheres augment in vivo bone formation in impaction bone grafting. Such microspheres with added biological functionality offer innovative therapeutic approaches to activate skeletal populations and enhance bone formation with reparative implications for hard tissues. PMID- 19135717 TI - Micromechanics of bone tissue-engineering scaffolds, based on resolution error cleared computer tomography. AB - Synchrotron radiation micro-computed tomography (SRmuCT) revealed the microstructure of a CEL2 glass-ceramic scaffold with macropores of several hundred microns characteristic length, in terms of the voxel-by-voxel 3D distribution of the attenuation coefficients throughout the scanned space. The probability density function of all attenuation coefficients related to the macroporous space inside the scaffold gives access to the tomograph-specific machine error included in the SRmuCT measurements (also referred to as instrumental resolution function). After Lorentz function-based clearing of the measured CT data from the systematic resolution error, the voxel-specific attenuation information of the voxels representing the solid skeleton is translated into the composition of the material inside one voxel, in terms of the nanoporosity embedded in a dense CEL2 glass-ceramic matrix. Based on voxel invariant elastic properties of dense CEL2 glass-ceramic, continuum micromechanics allows for translation of the voxel-specific nanoporosity into voxel-specific elastic properties. They serve as input for Finite Element analyses of the scaffold structure. Young's modulus of a specific CT-scanned macroporous scaffold sample, predicted from a Finite Element simulation of a uniaxial compression test, agrees well with the experimental value obtained from an ultrasonic test on the same sample. This highlights the satisfactory predictive capabilities of the presented approach. PMID- 19135718 TI - Formation of a human-derived fat tissue layer in P(DL)LGA hollow fibre scaffolds for adipocyte tissue engineering. AB - Development of adipose tissue-engineering strategies, where human bone marrow stromal cells (HBMSC) are combined with three-dimensional scaffolds, is likely to prove valuable for soft tissue restoration. In this study, we assessed the function of poly(DL-lactide-co-glycolide) (P(DL)LGA) hollow fibres in facilitating the development of HBMSC-derived adipocytes for advancement of an associated adipocyte layer. The large surface area of 75:25 P(DL)LGA fibres facilitated the rapid generation of extensive adipocyte aggregates from an undifferentiated HBMSC monolayer, where the fat-laden cells stained positive with Oil Red O and expressed the adipocyte marker, fatty acid binding protein 3 (FABP3). Following implantation subcutaneously in severely compromised immunodeficient mice, the adipogenic phenotype of the PLGA-adipocyte graft was maintained for up to 56 days. Confocal microscopy showed associated LipidTOX Deep Red neutral lipid staining in an (FL)P(DL)LGA fibre-adipocyte graft after 56 days, critical evidence demonstrating maintenance of the adipocyte phenotype in the subcutaneous graft. To support adipose tissue advancement in a defined volume, the P(DL)LGA-adipocyte scaffold was encapsulated within alginate/chitosan hydrogel capsules (typical diameters, 4.0 mm). In a 28-day in vivo trial in immunodeficient mice, clusters of the capsules were maintained at the subcutaneous site. An adipocyte tissue layer advancing within the surrounding hydrogel was demonstrated. PMID- 19135719 TI - Cellular automata model for drug release from binary matrix and reservoir polymeric devices. AB - Kinetics of drug release from polymeric tablets, inserts and implants is an important and widely studied area. Here we present a new and widely applicable cellular automata model for diffusion and erosion processes occurring during drug release from polymeric drug release devices. The model divides a 2D representation of the release device into an array of cells. Each cell contains information about the material, drug, polymer or solvent that the domain contains. Cells are then allowed to rearrange according to statistical rules designed to match realistic drug release. Diffusion is modeled by a random walk of mobile cells and kinetics of chemical or physical processes by probabilities of conversion from one state to another. This is according to the basis of diffusion coefficients and kinetic rate constants, which are on fundamental level just probabilities for certain occurrences. The model is applied to three kinds of devices with different release mechanisms: erodable matrices, diffusion through channels or pores and membrane controlled release. The dissolution curves obtained are compared to analytical models from literature and the validity of the model is considered. The model is shown to be compatible with all three release devices, highlighting easy adaptability of the model to virtually any release system and geometry. Further extension and applications of the model are envisioned. PMID- 19135720 TI - A nanoscale drug-entrapment strategy for hydrogel-based systems for the delivery of poorly soluble drugs. AB - The hydrophilic nature of hydrogel matrices makes them disadvantageous to entrap poorly soluble therapeutic agents and greatly restricts their applications as drug-delivery systems. In this study, we demonstrated that sustained delivery of lipophilic drugs in hydrogel-based devices can be readily achieved by enhancing retention of drugs within micelles. This nanoscale drug-entrapment strategy was applied to develop a polymeric drug-eluting stent. Sirolimus, a lipophilic anti proliferative/immunosuppressive drug, was entrapped into the hydrophobic core of Pluronic L121 micelles and then blended in a chitosan-based strip and crosslinked by an epoxy compound to fabricate test stents. It was found that the use of such a nanoscale drug-entrapment strategy was able to significantly increase the loading efficiency of lipophilic drugs, prevent the drug from aggregation and beneficially reduce its initial burst release; thus, the duration of drug release was extended considerably. When implanting the stent in rabbit infrarenal abdominal aortas, in-stent restenosis was markedly reduced and less inflammatory reaction was observed, while unfavorable effects such as delayed endothelial healing caused by the overdose of sirolimus could be significantly evaded. PMID- 19135722 TI - Arterial spin labeling shows cortical collateral flow in the endovascular treatment of vasospasm after post-traumatic subarachnoid hemorrhage. AB - We report here on the case of a patient who, 10 days prior to his admission to hospital, had suffered a bicycle accident. He presented with signs of minor dysphasia. A brain CT-scan revealed slight subarachnoid hemorrhage (SAH) in the left sylvian fissure as well as narrowing of the distal M1 segment of the left middle cerebral artery (MCA) on CT-angiography. MRI showed diffusion abnormalities and hypoperfusion in the left MCA territory with cortical hyperperfusion on arterial spin labeling (ASL). Arteriography confirmed the vasospasm, but showed no sign of aneurysm. Angioplasty of the narrowed MCA was successful, and follow-up MRI showed reperfusion of the MCA territory. PMID- 19135721 TI - Carvacrol is a novel inhibitor of Drosophila TRPL and mammalian TRPM7 channels. AB - Transient receptor potential (TRP) channels are essential components of biological sensors that detect changes in the environment in response to a myriad of stimuli. A major difficulty in the study of TRP channels is the lack of pharmacological agents that modulate most members of the TRP superfamily. Notable exceptions are the thermoTRPs, which respond to either cold or hot temperatures and are modulated by a relatively large number of chemical agents. In the present study we demonstrate by patch clamp whole cell recordings from Schneider 2 and Drosophila photoreceptor cells that carvacrol, a known activator of the thermoTRPs, TRPV3 and TRPA1 is an inhibitor of the Drosophila TRPL channels, which belongs to the TRPC subfamily. We also show that additional activators of TRPV3, thymol, eugenol, cinnamaldehyde and menthol are all inhibitors of the TRPL channel. Furthermore, carvacrol also inhibits the mammalian TRPM7 heterologously expressed in HEK cells and ectopically expressed in a primary culture of CA3-CA1 hippocampal brain neurons. This study, thus, identifies a novel inhibitor of TRPC and TRPM channels. Our finding that the activity of the non-thermoTRPs, TRPL and TRPM7 channels is modulated by the same compound as thermoTRPs, suggests that common mechanisms of channel modulation characterize TRP channels. PMID- 19135723 TI - Great genotypic and phenotypic diversities associated with copy-number variations of complement C4 and RP-C4-CYP21-TNX (RCCX) modules: a comparison of Asian-Indian and European American populations. AB - Inter-individual gene copy-number variations (CNVs) probably afford human populations the flexibility to respond to a variety of environmental challenges, but also lead to differential disease predispositions. We investigated gene CNVs for complement component C4 and steroid 21-hydroxylase from the RP-C4-CYP21-TNX (RCCX) modules located in the major histocompatibility complex among healthy Asian-Indian Americans (AIA) and compared them to European Americans. A combination of definitive techniques that yielded cross-confirmatory results was used. The medium gene copy-numbers for C4 and its isotypes, acidic C4A and basic C4B, were 4, 2 and 2, respectively, but their frequencies were only 53-56%. The distribution patterns for total C4 and C4A are skewed towards the high copy number side. For example, the frequency of AIA-subjects with three copies of C4A (30.7%) was 3.92-fold of those with a single copy (7.83%). The monomodular-short haplotype with a single C4B gene and the absence of C4A, which is in linkage disequilibrium with HLA DRB1*0301 in Europeans and a strong risk factor for autoimmune diseases, has a frequency of 0.012 in AIA but 0.106 among healthy European Americans (p=6.6x10(-8)). The copy-number and the size of C4 genes strongly determine the plasma C4 protein concentrations. Parallel variations in copy-numbers of CYP21A (CYP21A1P) and TNXA with total C4 were also observed. Notably, 13.1% of AIA-subjects had three copies of the functional CYP21B, which were likely generated by recombinations between monomodular and bimodular RCCX haplotypes. The high copy-numbers of C4 and the high frequency of RCCX recombinants offer important insights to the prevalence of autoimmune and genetic diseases. PMID- 19135724 TI - Characterization of an early signaling defect following Fc epsilonRI activation in the canine mastocytoma cell line, C2. AB - A comparison of IgE recognition by cognate receptors expressed on the C2 canine mastocytoma cell line with analogous events in a rat basophilic leukemia cell line transfected with the alpha-chain subunit of the canine high-affinity IgE receptor using flow cytometry show that canine IgE recognizes the alpha-chain of its cognate receptor on both cell lines. Our study confirms the expression of functional IgE receptors in both cell lines, but receptor-mediated signaling in the C2 line only supports the early stages of downstream signaling as shown by the phosphorylation of the gamma-chain and the failure to effect the phosphorylation of Syk. In contrast RBL-2H3 cells respond to sensitization with IgE and challenge with cognate antigen with tyrosine phosphorylation of the gamma subunits of the receptor complex followed by downstream phosphorylation of Syk and Ca(2+) mobilization, culminating in beta-hexosaminidase release. We propose that the identification of the precise signaling defect in C2 cells will yield useful information regarding the pathway leading to mast cell exocytosis and facilitate the restoration of the complete signaling cascade through complementation of the missing/defective signal transducer since signaling events downstream of Ca(2+) mobilization are intact as demonstrated by beta hexosaminidase release following non-immunologic stimulation with the calcium ionophore, A23187. PMID- 19135725 TI - Deletion of the G2A receptor fails to attenuate experimental autoimmune encephalomyelitis. AB - Lysophosphatidylcholine (LPC) is a chemotactic lysolipid produced during inflammation by the hydrolytic action of phospholipase A(2) enzymes. LPC stimulates chemotaxis of T cells in vitro through activation of the G protein coupled receptor, G2A. This has led to the proposition that G2A contributes to the recruitment of T cells to sites of inflammation and thus promotes chronic inflammatory autoimmune diseases associated with the generation and subsequent tissue infiltration of auto-antigen-specific effector T cells. However, one study suggests that G2A may negatively regulate T cell proliferative responses to antigen receptor engagement and thereby attenuates autoimmunity by reducing the generation of autoreactive T cells. To address the relative contribution of these G2A-mediated effects to the pathophysiology of T cell-mediated autoimmune disease, we examined the impact of G2A inactivation on the onset and severity of murine experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis (MS). Wild type (G2A(+/+)) and G2A-deficient (G2A(-/-)) C57BL/6J mice exhibited a similar incidence and onset of disease following immunization with MOG(35-55) peptide. Disease severity was only moderately reduced in G2A(-/-) mice. Similar numbers of MOG(35-55) specific T cells were generated in secondary lymphoid organs of MOG(35-55)-immunized G2A(+/+) and G2A(-/-) mice. Comparable numbers of T cells were detected in spinal cords of G2A(+/+) and G2A(-/-) mice. We conclude that the proposed anti-proliferative and chemotactic functions of G2A are not manifested in vivo and therefore therapeutic targeting of G2A is unlikely to be beneficial in the treatment of MS. PMID- 19135726 TI - Squamous carcinoma of the hypopharynx in children in Senegal: between disarray and enigma. AB - INTRODUCTION: The squamous carcinoma of the hypopharynx constitutes the first reason of hospitalization in our department. Different studies have been realized about it and showed an epidemiological profile characterized by the young age of the patients and the predominance of female. OBJECTIVES: To specify the place of the children in hypopharynx cancers. To evaluate the epidemiological criteria and the difficulties in the management. MATERIALS AND METHODS: Retrospective study of 11 years (1995-2005). Collection of the data concerning the epidemiology, clinical and paraclinical findings, the treatment, and the evolution was carried out at the University Department of Otorhinolaryngology in Dakar (Senegal). RESULTS: 15 charts of children have been collected. They presented in the majority of the cases, an advanced cancer, with 93% of T3T4. The lesion was localized at the retro-cricoid area in 4 cases, the pyriform sinus in 3 cases, the oesophagus junction in 2 cases, and the posterior wall in 1 case. In 5 cases the lesion was spreading to the totality of the hypopharynx. No case of smoking or alcohol has been noted. The anemia has been noted in 86.7% of the cases. A case of papillomatosis of the mouth and lips has been noted. The treatment was for most of cases as symptomatic, like tracheotomy with or without gastrostomy. The evolution was fatal with 11 deaths (73.33%). DISCUSSION: Our set is the richest set through the world of hypopharyngeal cancers in children. The Plummer Vinson syndrome noted in 86.7% of the cases and the viral infection by HPV could play a role in the genesis of this cancer. CONCLUSION: The cancer of the hypopharynx essentially affects the young women in Senegal without alcohol or tobacco consumption. It does not save the children, with an awful prognosis, however. So, it is imperative to organize a vast campaign of information of the populations on the bad prognosis of this cancer and to lead a large-scale epidemiological investigation, to get a better understanding of this cancer in our country. PMID- 19135727 TI - Contact in the genetics of autism and schizophrenia. AB - Although autism and schizophrenia are considered to be distinct neuropsychiatric developmental disorders, recent studies indicate that they share genetic factors. The same chromosomal rearrangements and several single genes have emerged as genetic risks in both disorders. One such gene is contactin-associated protein-2 (CNTNAP2). These findings raise the possibility that these neuropsychiatric disorders share pathogenic mechanisms and that similar defects in biological pathways of brain development might underlie the phenotypic spectrum of these disorders. PMID- 19135728 TI - Purinergic signalling in inflammation of the central nervous system. AB - Inflammation is the most fundamental body reaction to noxious stimuli. No vascularized tissue, organ or apparatus is free from this response. Several mediators of inflammation, originating from outside (exogenous) or inside (endogenous) the body, are known. Among the endogenous factors, extracellular nucleotides and nucleosides are attracting interest for their ubiquity and striking ability to modulate diverse immune responses. Until recently, it was doubted that the central nervous system (CNS), reportedly an 'immunoprivileged organ', could be the site of immune reactions. Nowadays, it is acknowledged that inflammation and immunity have a key role in a vast range of CNS diseases. Likewise, it is clear that purinergic signalling profoundly affects neuroinflammation. Here, we provide a brief update of the state of the art in this expanding field. PMID- 19135729 TI - Deciphering complex mechanisms in neurodegenerative diseases: the advent of systems biology. AB - Classical biological methods involving analyses of one or several genes have been the mainstay for studying the pathogenesis of neurodegenerative disorders. However, it has become clear that these diseases exhibit complex molecular interactions involving both host genomes and environmental determinants. Systems biology represents an integrated and deeper investigation of interacting biomolecules within cells or organisms. This approach has only recently become feasible as high-throughput technologies including cDNA microarrays, mass spectrometric analyses of proteins and lipids together with rigorous bioinformatics have evolved. Herein, we review recent developments from studies of systems biology applied to multiple sclerosis, Alzheimer's disease and HIV associated dementia as three prototypic neurodegenerative disorders. Existing high-content data derived from clinically and experimentally derived neural tissues point to convergent pathways among these neurodegenerative disorders, which transcend descriptive studies to reach a more integrated understanding of disease pathogenesis and, in some instances, highlighting 'druggable' network nodes. PMID- 19135730 TI - Remote neuroimmune signaling: a long-range mechanism of nociceptive network plasticity. AB - Chronic pain secondary to neuronal injury is actively and continuously modulated at multiple locations along the sensory neuraxis. Here, we describe how nociceptive neurons of the spinal cord and thalamus process and communicate nociceptive information in terms of precisely calibrated firing patterns. We then discuss how several cell types with immunogenic properties (e.g. blood cells and glia) cause system-wide interference in nociceptive processing through novel signaling schema, thus contributing to nociceptive network plasticity and chronic pain. PMID- 19135731 TI - Calculated serum calcium is an insufficient surrogate for measured ionized calcium. AB - Direct measurement of serum-ionized calcium (Ca2+) is the best available method to assess 'true' calcemia because serum total calcium is confounded by serum proteins, mainly albumin. However, calculated surrogates for Ca2+, albumin adjustment particularly, have been widely used in the literature despite their insufficiency to evaluate calcemic status accurately. Recently, a new formula, in which serum total protein instead of albumin is used for adjustment of total calcium, has been suggested for very old hospitalized patients. We compared these two surrogates for Ca2+ with the direct measurement in both frail bedridden inpatients and in a general aged population. Both surrogates were equally insufficient in detecting hypocalcemia in both patient groups. However, the sensitivity and specifity for detecting hypercalcemia by both surrogates were relatively high in the bedridden inpatients. Precise assessment of calcemic status is particularly needed in research and in detecting mild hypo- or hypercalcemia. However, the calculated surrogates for Ca2+ have consistently failed in accurate assessment of 'true' calcemia. Thus, the direct measurement of Ca2+ should be favored instead of readily available automated surrogates. Adjustments of calcium for albumin or total protein may be useful in clinical practice in excluding hypercalcemia in very old bedridden patients, only. PMID- 19135732 TI - Successful use of endothelial progenitor cell capture stent for treatment of left main coronary artery disease before non-cardiac surgery for abdominal aortic aneurysm. AB - We report on the novel use of endothelial progenitor cell (EPC) capture stent for the treatment of left main coronary artery disease (CAD) for a patient who required a high risk non-cardiac surgery for his rapidly expanding abdominal aortic aneurysm (AAA). Instead of using bare metal stent, we decided to use the EPC capture stent to treat his left main disease as it had a unique "pro-healing" properties. We were able to reduce his peri-operative risk and he underwent the AAA surgery successfully with no adverse events. To our knowledge, this is the first reported case of the successful use of EPC capture stent for the treatment of left main CAD before a high risk non-cardiac surgery for AAA. PMID- 19135733 TI - Predictors of coronary spasm in patients with chest pain and no significant coronary stenosis. PMID- 19135734 TI - Ergotamine-induced cardiovascular toxicity: mechanisms and clinical significance. PMID- 19135735 TI - Atypical basal type takotsubo cardiomyopathy: MDCT findings correlated with echocardiography. AB - Takotsubo cardiomyopathy is characterised by reversible left ventricular apical ballooning and no significant coronary artery stenosis. A new variant, a mid ventricular pattern of takotsubo cardiomyopathy, has been reported. However, there have been few reports on basal wall akinesia or hypokinesia with preservation of the apical and mid-ventricular wall motion shown at echocardiography and ventriculography. We report the case of a 42-year-old patient who presented with atypical basal type takotsubo cardiomyopathy and recovered fully within 2 weeks. PMID- 19135736 TI - Further comments on oral transmission of Chagas' disease in Brazil: epidemiology, geographical distribution and viability of the infective parasite. AB - In 2006, Brazilian government received the international certificate of interruption of the vectorial transmission of Chagas' disease. However, outbreaks reported in Brazilian Amazon rainforest bear a regular occurrence and represents a relevant regional epidemiological gauge. The wild life cycle of the Chagas' disease transmission (i.e., triatomine-marsupial cycle) is present outside the previously reported endemic belt, ubiquitously, as infective triatomines can be demonstrated in Palm trees widespread all over the Amazon rainforest. As humans invade the rainforest, one is incidentally caught up and further becomes' an active part of American trypanosomiasis wild life cycle. PMID- 19135737 TI - Thorough search for myopathy increases the prevalence of noncompaction. PMID- 19135738 TI - A 31 year old woman with essential hypertension grade III and branch retinal vein occlusion with homozygous C677T MTHFR hyperhomocysteinemia and high Lp(a) levels. AB - We report a 31-year old woman with essential hypertension grade III and history of branch retinal vein occlusion in the setting of hyperhomocysteinemia due to homozygous MTHFR gene mutation and elevated Lp(a). The patient was treated successfully with antihypertensive treatment, acetylsalicylic acid and multivitamin complex supplementation. PMID- 19135739 TI - Prolonged cardiogenic shock caused by a high-dose intravenous administration of dopamine in a patient with takotsubo cardiomyopathy. AB - We report a patient with takotsubo cardiomyopathy in whom cardiogenic shock continued with a high intraventricular pressure gradient (IVPG) under a high-dose intravenous administration of dopamine and in whom cessation of the administration improved the hemodynamics dramatically. The present case report suggests that the administration of dopamine may lead to further hemodynamic deterioration in patients with takotsubo cardiomyopathy who exhibits an IVPG. PMID- 19135740 TI - Anthracycline-induced cardiomyopathy: favourable effects of cardiac resynchronization therapy. AB - We report a case of severe refractory congestive heart failure after anthracycline chemotherapy in a patient with a narrow QRS interval on the electrocardiogram and echocardiographic evidence of left ventricular dyssynchrony, where cardiac resynchronization therapy resulted in normalization of left ventricular ejection fraction and marked symptomatic relief. PMID- 19135741 TI - Oral potassium supplement use and outcomes in chronic heart failure: a propensity matched study. AB - BACKGROUND: Hypokalemia is common in heart failure (HF) and is associated with increased mortality. Potassium supplements are commonly used to treat hypokalemia and maintain normokalemia. However, their long-term effects on outcomes in chronic HF are unknown. We used a public-use copy of the Digitalis Investigation Group (DIG) trial dataset to determine the associations of potassium supplement use with outcomes using a propensity-matched design. METHODS: Of the 7788 DIG participants with chronic HF, 2199 were using oral potassium supplements at baseline. We estimated propensity scores for potassium supplement use for each patient and used them to match 2131 pairs of patients receiving and not receiving potassium supplements. Matched Cox regression models were used to estimate associations of potassium supplement use with mortality and hospitalization during 40 months of median follow-up. RESULTS: All-cause mortality occurred in 818 (rate, 1327/10,000 person-years) and 802 (rate, 1313/10,000 person-years) patients respectively receiving and not receiving potassium supplements (hazard ratio {HR} when potassium supplement use was compared with nonuse, 1.05; 95% confidence interval {CI}, 0.94-1.18; P=0.390). All-cause hospitalizations occurred in 1516 (rate, 4777/10,000 person-years) and 1445 (rate, 4120/10,000 person-years) patients respectively receiving and not receiving potassium supplements (HR, 1.15; 95% CI, 1.05-1.26; P=0.004). HRs (95% CI) for hospitalizations due to cardiovascular causes and worsening HF were respectively 1.19 (95% CI, 1.08-1.32; P=0.001) and 1.27 (1.12-1.43; P<0.0001). CONCLUSION: The use of potassium supplements in chronic HF was not associated with mortality. However, their use was associated with increased hospitalization due to cardiovascular causes and progressive HF. PMID- 19135742 TI - Recurrent acute pulmonary embolism and paroxysmal atrial fibrillation associated with subclinical hyperthyroidism. AB - Subclinical hyperthyroidism is an increasingly recognized entity that is defined as a normal serum free thyroxine and free triiodothyronine levels with a thyroid stimulating hormone level suppressed below the normal range and usually undetectable. It has been reported that subclinical hyperthyroidism is not associated with coronary heart disease or mortality from cardiovascular causes but it is sufficient to induce arrhythmias including atrial fibrillation and atrial flutter. It has also been reported that increased factor X activity in patients with subclinical hyperthyroidism represents a potential hypercoagulable state. Elevated D-dimer levels have been reported in acute pulmonary embolism and it has also been reported that right ventricular overload and hypoxia in acute pulmonary embolism may lead to right ventricular myocardium injury reflected by elevated cardiac troponin levels too. We present a case of recurrent pulmonary embolism associated with subclinical hyperthyroidism, in an 81-year-old Italian woman. Also this case focuses attention on the importance of a correct evaluation of subclinical hyperthyroidism. PMID- 19135743 TI - A case with the longest angiographically documented left main coronary artery. AB - The left main coronary artery (LMC) is generally considered to be between 5 and 15 mm long. We report a case with the longest angiographically documented LMC (41 mm) in a patient with acute inferior myocardial infarction. PMID- 19135744 TI - Acute efficacy of low-dose human atrial natriuretic peptide monotherapy without loop diuretics for acute decompensated heart failure with left ventricular systolic dysfunction: a case report. AB - A 64-year-old man was admitted to our hospital with new-onset acute decompensated heart failure (ADHF). He had left ventricular systolic dysfunction and chronic atrial fibrillation with a rapid ventricular response. Initial treatment with low dose recombinant human atrial natriuretic peptide (hANP) alone and no loop diuretics had an immediate effect on ADHF and left ventricular systolic dysfunction. This case demonstrates that low-dose hANP (0.01 microg/kg/min) monotherapy can be safe and effective for ADHF, with no loop diuretics being required during hospitalization. However, clinical trials will be needed to further evaluate the acute efficacy and safety of hANP monotherapy in patients with ADHF. PMID- 19135745 TI - Dual atrioventricular nodal physiology in young child. AB - Dual atrioventricular nodal physiology has been reported. Atrioventricular nodal reentrant tachycardia is the most common supraventricular tachycardia associated with dual atrioventricular nodal physiology. We present a case of dual atrioventricular nodal physiology in a 12-year-old Italian boy. PMID- 19135746 TI - The first report of late stent thrombosis leading to acute myocardial infarction in patient receiving the new endothelial progenitor cell capture stent. AB - The endothelial progenitor cell (EPC) capture stent is a stainless steel coronary stent covered with antibodies specific to EPC's surface antigens, designed to promote the faster formation of a functional endothelial lining. This may prevent both stent thrombosis and restenosis as well as reduce the need of prolonged double antiplatelet therapy. Here we report a case of late (156 days after implantation) EPC capture stent thrombosis and re-stenosis of a second EPC capture stent occurring both in the same patient after withdrawal of clopidogrel. Intravascular ultrasound performed during his acute admission showed undersizing of the stent with thrombosis. The present case raises the question about the safety of the EPC capture stent and the need to prolong at least for 6 months double antiplatelet therapy also in patients receiving this new stent. PMID- 19135747 TI - Nanotechnology and its applications in the food sector. AB - Nanoscience and nanotechnology are new frontiers of this century. Their applications to the agriculture and food sector are relatively recent compared with their use in drug delivery and pharmaceuticals. Smart delivery of nutrients, bioseparation of proteins, rapid sampling of biological and chemical contaminants and nanoencapsulation of nutraceuticals are some of the emerging topics of nanotechnology for food and agriculture. Advances in technologies, such as DNA microarrays, microelectromechanical systems and microfluidics, will enable the realization of the potential of nanotechnology for food applications. In this review, we intended to summarize the applications of nanotechnology relevant to food and nutraceuticals together with identifying the outstanding challenges. PMID- 19135748 TI - Inter-professional variability in the assignment and recording of acute toxicity grade using the RTOG system during prostate radiotherapy. AB - BACKGROUND AND PURPOSE: To compare the routine acute toxicity documentation practices of therapists and oncologists using the RTOG lower GI and GU scales. METHODS AND MATERIALS: Ninety consecutive prostate radiotherapy patients were identified. The weekly urinary and rectal acute toxicity grades routinely documented by therapists and oncologists were collected retrospectively from radiotherapy charts. These data were paired together, and compared between the professional groups. RESULTS: Only RTOG acute toxicity grades between 0 and 2 were recorded by either group. The overall rate of documentation was high (97% therapists/86% oncologists), but the rate of quantitative documentation was low from the oncologists (46%) who used a free-form text field for recording purposes. There was no significant difference in the incidence of maximum grade of acute toxicity reported by either professional group (p>0.1). There was good RTOG score concordance between the observer groups (kappa=0.756), with pair-wise absolute agreement in 76%. Pair-wise discrepancies between the observers were commonly attributable to differences in the time/date of assessment. CONCLUSIONS: Despite some methodological limitations, this study found that therapist-assessed RTOG acute toxicity grades demonstrated a good level of agreement with the grades assigned by their oncologist colleagues. PMID- 19135749 TI - Dose escalation in the radiotherapy of non-small-cell lung cancer with aperture based intensity modulation and photon beam energy optimization for non preselected patients. AB - PURPOSE: To verify the potential of aperture-based intensity-modulated radiotherapy (AB-IMRT) to realize dose escalation plans for non-preselected non small-cell lung cancer (NSCLC) patients, using photon beam energy optimization. METHODS AND MATERIALS: Seven cases of NSCLC were retrospectively studied. Clinical reference plans were made at 60 Gy by an experienced dosimetrist. Dose escalation was applied to PTV2, a subvolume within the main PTV1. Escalation plans were optimized by considering beam angles (table and gantry), energy (6 and 23 MV) and weights, for an increasing dose to the PTV2, starting from 66 Gy and keeping 30 fractions. RESULTS: In five cases, doses over 78 Gy could be achieved before exceeding organs at risk (OARs) standard tolerance. Peripheral overdosages, as well as lung and spinal cord tolerance doses, limited escalation. Means+/-SD V(95%) parameters were (97.3+/-0.9)% for PTV1s and (96.7+/-2.2)% for PTV2s. Doses to OARs were also maintained at acceptable levels. Optimized plans made use of both low- and high-energy beams and had a similar number of monitor units compared to the 60 Gy clinical plans. CONCLUSIONS: The AB-IMRT system can successfully realize dose escalation for a sizeable number of cases. Plans produced contained few large segments, and are applicable to a wide range of tumor volumes and locations. PMID- 19135750 TI - Stereotactic radiosurgery for the treatment of brain metastases. AB - BACKGROUND: The objective of this systematic review was to summarise the current evidence concerning radiosurgical treatment (SRS) of newly diagnosed brain metastasis and to compare SRS as a single or additional treatment to treatment alternatives with regard to medical effectiveness and safety. METHODS: A structured search of electronic databases was performed to identify relevant publications from 2002 through 2007. Studies targeting patients with brain metastases were included. Standardised quality assessment and data extraction were performed. RESULTS: Of 1496 publications, 16 studies were included. The mean survival in most studies was less than 12 months. There was evidence that SRS plus WBRT was associated with improved local tumour control and neurological functioning compared to either treatment alone. Only in patients with single metastasis, this resulted in improved survival. There was inconclusive evidence when comparing SRS to WBRT, Neurosurgery (NS) or Hypofractionated Radiotherapy (HCSRT). The Quality of life (Qol) was not investigated. CONCLUSION: SRS plus WBRT was associated with improved local tumour control and neurological functioning compared to either treatment alone. Only in certain patients, this resulted in improved survival. Methodologically rigorous studies are therefore warranted to investigate further treatment options, and in view of the poor prognosis, to investigate Qol and neurological functioning. PMID- 19135751 TI - Favorable outcomes of radiotherapy for early-stage mucosa-associated lymphoid tissue lymphoma. AB - PURPOSE: The aim of this study was to evaluate the efficacy of radiation therapy (RT) for early-stage mucosa-associated lymphoid tissue (MALT) lymphoma. MATERIALS AND METHODS: Patients with stage IotaE (n=48) and stage capital PE, CyrillicE (n=2) MALT lymphoma treated with RT were reviewed. The primary tumor originated in the stomach in 20 patients, in the orbit in 9 patients, in the conjunctiva or eyelid and the parotid glands in 6 patients each, and 9 patients in the others. The median total RT dose was 32Gy (range, 25.6-50Gy). The median follow-up time was 50 months. RESULTS: Although disease did not recur in the RT field in any patient regardless of the total dose, disease recurred outside the RT field in the seven patients. As all recurrences were localized, salvage RT was performed for each recurrence and achieved complete response without recurrence in the field. The 5-year overall survival, local control, and progression-free survival rates were 96.6%, 100%, and 82.2%, respectively. CONCLUSIONS: A total dose of 25 30Gy is appropriate for local control of MALT lymphoma. RT is also an effective salvage therapy in cases of localized recurrence. PMID- 19135752 TI - Two related cases of type A insulin resistance with compound heterozygous mutations of the insulin receptor gene. AB - We describe two sisters with type A insulin resistance. In contrast to common situation for this genetic disorder, the sisters harbored compound heterozygous mutations in the insulin receptor gene associated with mild glucose intolerance. The cases highlight the diversity of clinical phenotypes associated with mutations of the insulin receptor gene. PMID- 19135753 TI - Risperidone-associated newly diagnosed diabetes and fatal diabetes ketoacidosis in a young schizophrenic patient. AB - A 27-year-old man, who has been using risperidone for two months as the treatment for schizophrenia, with no previous history of diabetes was admitted to the hospital with the presentation of severe diabetes ketoacidosis and subsequent fatal progression. PMID- 19135754 TI - Resistance training improves metabolic health in type 2 diabetes: a systematic review. AB - This paper systematically reviews the effect of resistance training (RT) on glycemic control and insulin sensitivity in adults with type 2 diabetes. Twenty studies were included, with the volume, frequency and intensity of RT varying markedly. Supervised RT improved glycemic control and insulin sensitivity, however, when supervision was removed compliance and glycemic control decreased. Evidence indicates the mechanisms behind the improvements to glucose tolerance require further elucidation. Although research demonstrates apparent benefits of RT for individuals with diabetes, further research is required to elucidate the minimum effective dose by describing frequency, intensity and the duration of acute and chronic improvements. PMID- 19135755 TI - Glycemic control and survival of diabetic hemodialysis patients--importance of lower hemoglobin A1C levels. AB - AIMS: The significance of hemoglobin A1C (HbA1C) on the survival of diabetic hemodialysis patients still remains controversial. We investigated the impact of HbA1C on the survival. METHODS: A total of 122 diabetic patients on maintenance hemodialysis (age, 59.9+/-11.9 years [mean+/-SD]; hemodialysis duration: 53+/-38 months) were surveyed (survey period: 46+/-19 months). RESULTS: The cumulative survival of the poor glycemic control group (mean HbA1C of 3-month period > or =6.3%, n=62) was significantly lower than that of the good group (HbA1C<6.3%, n=60), as determined by Kaplan-Meier estimation (P=0.0084, log-rank test). Kaplan Meier analysis also demonstrated that both cardiovascular and non-cardiovascular mortalities were higher in the poor group than in the good group (P=0.0545 and P=0.0453, respectively). In a multivariate Cox proportional hazard model, the mean HbA1C was a significant predictor of survival (OR 1.260 per 1.0%, 95% CI 1.020-0.579, P=0.0325). CONCLUSIONS: Poor glycemic control is an independent predictor of poor prognosis in diabetic hemodialysis patients. HbA1C is a clinically useful parameter for identifying the risk for mortality, both for cardiovascular and non-cardiovascular mortality, and that careful management of glycemic control by use of HbA1C is important. PMID- 19135756 TI - Cost-effectiveness analyses of drug eluting stents versus bare metal stents: a systematic review of the literature. AB - OBJECTIVES: Drug eluting stents (DES) used to treat coronary lesions reduce rates of in-stent restenosis and therefore the need for repeat revascularization compared to bare metal stents (BMS). DES, however, are considerably more expensive than BMS. We evaluated whether DES are a cost-effective alternative for BMS. METHODS: Reports of Health Technology Assessment agencies were assessed and a systematic review of economic evaluations comparing DES with BMS was performed. RESULTS: Nineteen full economic evaluations were identified. Some studies indicate that DES may be cost-effective or even cost-saving in specific patients, when used for coronary lesions with a high propensity of restenosis such as long lesions, lesions in narrow vessels, or in patients with diabetes. Other studies mention DES is not cost effective at all with ICERs of more than 200,000 Canadian dollar per QALY gained. One of the main determining factors is the influence of protocol mandated angiographic follow-up in RCTs. The risk for a re-intervention using BMS ranges from 5% to 14% in registries and is much smaller than reported in RCTs (up to 30%). As a result, the absolute reduction in repeat revascularization by DES compared to BMS is smaller in real life. Moreover, using DES instead of BMS does not increase survival or decrease myocardial infarctions. The combination of (a) a higher cost (>euro700) for DES versus BMS; (b) no life years gained; (c) a relatively small absolute reduction in repeat procedures; and (d) a small improvement in QoL for a short period (less than 0.15 during the first month after the re-intervention), results in unfavourable cost effectiveness ratios. CONCLUSIONS: Although several studies conclude that DES may be cost effective in large subgroups of patients, under real-world conditions, the cost-effectiveness of DES is unfavourable in comparison with BMS. PMID- 19135757 TI - Evaluation of lesions corresponding to ground-glass opacities that were resected after computed tomography follow-up examination. AB - BACKGROUND: Ground-glass opacity (GGO), which is closely related with bronchioloalveolar carcinoma (BAC), is being detected more frequently. BAC is considered to be a relatively less aggressive tumor, and immediate resection at the time of detection might not be necessary. Therefore, when GGO is detected, a CT follow-up examination is often performed. If growth is detected during the follow-up CT examination, resection is usually considered. However, the possible treatment delay caused by the scheduling of a CT follow-up examination is an issue that must be clarified. Since the cancer might progress during the follow up period, such follow-up periods might have a negative influence on the patient's prognosis. This study attempted to clarify whether CT follow-up causes treatment delay. METHODS: A total of 113 lung cancer patients with pure or mixed GGO findings who underwent a resection after a CT follow-up examination between 1999 and 2005 were retrospectively examined. The CT findings at the initial detection, the changes in the CT findings during the CT follow-up period, the histology, the pathological stage and the outcomes after resection were reviewed and evaluated. RESULTS: The CT finding at the time of the initial detection showed pure GGO in 63 patients and mixed GGO in 50 patients. Histology revealed that adenocarcinoma was found in all 113 patients; squamous cell carcinoma was not found in any of the patients. One-hundred twelve patients were diagnosed as having Stage IA, and a singe patient with visceral pleura invasion was diagnosed as having Stage IB. Complete resections were performed in all the patients. The median postoperative follow-up period was 45.0 months. No recurrences or deaths were observed during the study period. CONCLUSIONS: No treatment delays or negative influences on patient outcome resulted from the CT follow-up period. A future prospective study should be conducted to establish the optimal CT follow up program. PMID- 19135758 TI - Effect of highly active antiretroviral therapy on survival of HIV infected patients with non-small-cell lung cancer. AB - OBJECTIVE: To evaluate the impact of highly active antiretroviral therapy (HAART) on survival in HIV infected patients with non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: All consecutive HIV infected patients with NSCLC diagnosed between 06/1996 and 03/2007 at two University hospitals in Paris (France) were prospectively followed until death. The association between survival and clinical and biological factors was analyzed by univariate and multivariate models. Survival analysis was performed by Kaplan-Meier estimates and the Cox proportional hazards regression model. RESULTS: During the study period, NSCLC was diagnosed in 49 consecutive HIV infected patients (median age 46 years); 84% had advanced disease. Median survival was 8.1 months (range 5-10 months). In multivariate analysis, baseline parameters with significant positive impact on survival included performance status (PS) < or =1 (HR=0.2, 95%CI [0.09, 0.46], p=0.0001), stage I-II disease (HR=0.15, 95%CI [0.04, 0.53], p=0.003), and use of HAART (HR=0.4, 95%CI [0.2, 0.9], p=0.027). CONCLUSION: HAART is a good prognostic factor for survival in HIV infected patients with NSCLC. Stage of disease and PS are two other valid survival prognostic factors. PMID- 19135759 TI - Houston's medical disaster response to Hurricane Katrina: part 2: transitioning from emergency evacuee care to community health care. AB - After Hurricane Katrina hit the Gulf Coast on August 29, 2005, thousands of ill and injured evacuees were transported to Houston, TX. Houston's regional disaster plan was quickly implemented, leading to the activation of the Regional Hospital Preparedness Council's Catastrophic Medical Operations Center and the rapid construction of a 65-examination-room medical facility within the Reliant Center. A plan for triage of arriving evacuees was quickly developed and the Astrodome/Reliant Center Complex mega-shelter was created. Herein, we discuss major elements of the regional disaster response, including regional coordination, triage and emergency medical service transfers into the region's medical centers, medical care in population shelters, and community health challenges. PMID- 19135760 TI - Houston's medical disaster response to Hurricane Katrina: part 1: the initial medical response from Trauma Service Area Q. AB - After Hurricane Katrina hit the Gulf Coast on August 29, 2005, thousands of ill and injured evacuees were transported to Houston, TX. Houston's regional disaster plan was quickly implemented, leading to the activation of the Regional Hospital Preparedness Council's Catastrophic Medical Operations Center and the rapid construction of a 65-examination-room medical facility within the Reliant Center. A plan for triage of arriving evacuees was quickly developed and the Astrodome/Reliant Center Complex mega-shelter was created. Herein, we discuss major elements of the regional disaster response, including regional coordination, triage and emergency medical service transfers into the region's medical centers, medical care in population shelters, and community health challenges. PMID- 19135761 TI - Marked variability in adherence to hand hygiene: a 5-unit observational study in Tuscany. AB - BACKGROUND: International authorities recommend that the hand hygiene of health care workers be improved to prevent health care-associated infection. In 2005, Tuscany, a region in central Italy, initiated a campaign to improve hand hygiene that focused on raising awareness and educating health care workers. We assessed hand hygiene rates approximately 3 years after the campaign was initiated in 5 units of 2 hospitals in Florence, Italy, the capital of Tuscany. We also were curious whether variability would exist in the hand hygiene rates despite the close proximity of the units. METHODS: We conducted a 3-month observational study in 2008 to assess hand hygiene adherence of doctors and nurses. Four of the units (ophthalmology, cardiology, geriatrics, and infectious diseases) were within one hospital, and the fifth unit (an emergency department) was in another hospital located less than 1 km away. External observers were used to assess the hand hygiene adherence of doctors and nurses before patient contact. RESULTS: A total of 665 doctor-patient observations and 1147 nurse-patient observations were made. Doctors used some type of hand hygiene before touching the patient in 28% of their patient interactions (soap and water in 16% and alcohol-based handrub in 12%). Nurses used some type of hand hygiene in 34% of their interactions (soap and water in 27% and alcohol-based handrub in 7%). Hand hygiene adherence varied substantially across the units, from a low of 6% to a high of 66% for doctors and from 19% to 56% for nurses. The correlation between nurse adherence and doctor adherence was 0.90. CONCLUSION: The overall rates of hand hygiene adherence observed were similar to those found when Tuscany initiated a hand hygiene campaign 3 years earlier. Focusing on overall rates may be misleading, however, because substantial variability existed between units. Furthermore, these rates come only from the "first moment" (before touching the patient) and can only be compared with rates from studies using the same approach. PMID- 19135762 TI - Myocardial (123)I-MIBG scintigraphy for differentiation of Lewy bodies disease from FTD. AB - Clinical distinction between Lewy bodies disease (LBD) and frontotemporal dementia (FTD) is sometimes difficult. Nigrostriatal dopaminergic degeneration occurs in both LBD and FTD, limiting helpfulness of DAT imaging to differentiate these forms of dementia. Several studies have emphasized the usefulness of myocardial scintigraphy with (123)Metaiodobenzylguanidine ((123)I-MIGB) in assessing the sympathetic nerve terminals in LBD demonstrating that cardiac (123)I-MIGB uptake is decreased in patients with this disease. We investigated the role of cardiac (123)I-MIBG scintigraphy in differentiating patients with LBD from those with FTD. Clinical diagnosis of LBD and FTD was determined according to established consensus criteria. Nine patients with LBD (1 possible and 8 probable), 6 patients with FTD, and 16 control subjects were involved in the study. The heart to mediastinum ratio (H/M) of (123)I-MIBG uptake was markedly reduced in all patients with LBD (H/M early: 1.25+/-0.12; delayed: 1.14+/-0.13) whereas it was normal in patients with FTD (H/M early: 1.86+/-0.20; delayed: 1.80+/-0.23) and in controls (H/M early: 1.91+/-0.17; delayed: 1.99+/-0.19), suggesting that cardiac (123)I-MIBG scintigraphy can help distinguish patients with LBD from those with FTD. PMID- 19135764 TI - Persistent toxic substances in remote lake and coastal sediments from Svalbard, Norwegian Arctic: levels, sources and fluxes. AB - Surface sediments from remote lakes and coastal areas from Ny-Alesund, Svalbard, Norwegian Arctic were analyzed for polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs). Relatively high levels of PAHs were encountered from several lakes from Ny-Alesund, which were within the range of levels reported for European high mountain lakes and some urban/industrialized areas in the world, pointing to the role of remote Arctic lakes as potential reservoir of semi-volatile organic compounds. Specific patterns of PBDEs were observed, showing higher concentrations of lower brominated compounds such as BDE-7, 17 and 28. Estimated surface sediment fluxes of PAHs in Ny-Alesund remote lakes were similar to those observed for some European high mountain lakes. The current PAH levels in sediments from three lakes exceeded Canadian sediment quality guidelines, suggesting the presence of possible risks for aquatic organisms and the need for further studies. PMID- 19135763 TI - A diaryl sulfide, sulfoxide, and sulfone bearing structural similarities to combretastatin A-4. AB - Studies examining various spacer groups that link the two aromatic rings of combretastatin A-4 (CA4) have shown that the biological activity of analogs does not require the cis-stilbene configuration of CA4. Oxygen or nitrogen, carbonyl, methylene and ethylene spacers, for example, are present in CA4 analogs that show good activity. Up to now sulfur was not tested for this purpose. In this article we describe the synthesis of sulfide, sulfoxide and sulfone spacers between two aromatic rings comparable to those of CA4. We also compared them with CA4 for inhibitory effects on cell growth, tubulin polymerization, and the binding of [(3)H]colchicine to tubulin. We found that the sulfide is highly active and may be a lead compound for the preparation of antitumor compounds. PMID- 19135765 TI - Inorganic arsenic speciation at river basin scales: the Tinto and Odiel rivers in the Iberian Pyrite Belt, SW Spain. AB - The Tinto and Odiel rivers are heavily affected by acid mine drainage from mining areas in the Iberian Pyrite Belt. In this work we have conducted a study along these rivers where surface water samples have been collected. Field measurements, total dissolved metals and Fe and inorganic As speciation analysis were performed. The average total concentration of As in the Tinto river (1975 microg L(-1)) is larger than in the Odiel river (441 microg L(-1)); however, the mean concentration of As(III) is almost four times higher in the Odiel. In wet seasons the mean pH levels of both rivers (2.4 and 3.2 for the Tinto and Odiel, respectively) increase slightly and the amount of dissolved total arsenic tend to decrease, while the As(III)/(V) ratio strongly increase. Besides, the concentration of the reduced As species increase along the water course. As a result, As(III)/(V) ratio can be up to 100 times higher in the lower part of the basins. An estimation of the As(III) load transported by both rivers into the Atlantic Ocean has been performed, resulting in about 60 kg yr(-1) and 2.7t yr( 1) by the Tinto and Odiel rivers, respectively. PMID- 19135766 TI - A steady state visually evoked potential investigation of memory and ageing. AB - Old age is generally accompanied by a decline in memory performance. Specifically, neuroimaging and electrophysiological studies have revealed that there are age-related changes in the neural correlates of episodic and working memory. This study investigated age-associated changes in the steady state visually evoked potential (SSVEP) amplitude and latency associated with memory performance. Participants were 15 older (59-67 years) and 14 younger (20-30 years) adults who performed an object working memory (OWM) task and a contextual recognition memory (CRM) task, whilst the SSVEP was recorded from 64 electrode sites. Retention of a single object in the low demand OWM task was characterised by smaller frontal SSVEP amplitude and latency differences in older adults than in younger adults, indicative of an age-associated reduction in neural processes. Recognition of visual images in the more difficult CRM task was accompanied by larger, more sustained SSVEP amplitude and latency decreases over temporal parietal regions in older adults. In contrast, the more transient, frontally mediated pattern of activity demonstrated by younger adults suggests that younger and older adults utilize different neural resources to perform recognition judgements. The results provide support for compensatory processes in the aging brain; at lower task demands, older adults demonstrate reduced neural activity, whereas at greater task demands neural activity is increased. PMID- 19135768 TI - Lack of toxicity by medium chain triglycerides (MCT) in canines during a 90-day feeding study. AB - Dietary fats in food are natural energy sources to animals and are included in the American Association of Feed Control Officials (AAFCO) manual as a requirement for dog food. Medium chain triglycerides are comprised of a glycerol backbone esterified to medium chain length (8-12 carbon) fatty acids (FA) and, in the context of this report, are all saturated FA. Unlike esterified long chain (>12 carbons) FA (long chain triglycerides or LCT), MCT are lower in caloric value, and are eliminated from the body more quickly than LCT. The objective of this study was to determine the safety of MCT when fed to beagles for 90 days at levels of 0%, 5%, 10%, and 15% MCT added to conventional feed. The beagles were monitored for signs of toxicity by clinical observations, body weight measurements, food consumption level, physical examinations, hematology and serum chemistry, ophthalmic examinations, and urinalysis. There were no signs of toxic effects observed in any of the animals that were related to feed, and the animal viability was 100% at the end of the study. Some animals exhibited significant increased blood urea nitrogen, potassium and cholesterol levels in the 10% and 15% MCT-fed groups. Also, in the same groups with elevated nitrogen, there were concomitant reductions in total blood protein and urine volumes. These changes in serum chemistry may be the result of protein sparing effects due to the high levels of MCT intake, and are not deemed to be pathological in nature. Animals receiving 15% MCT in feed had lower levels of food intake due to palatability issues. From the other examination parameters, there were no significant changes noted between groups receiving MCT and vehicle feed. No safety concerns were noted at any dose level, although an issue with palatability precluded identifying 15% as the highest dose level tested. PMID- 19135767 TI - Brain mechanisms associated with background monitoring of the environment for potentially significant sensory events. AB - Background monitoring is a necessary prerequisite to detect unexpected changes in the environment, while being involved in a primary task. Here, we used fMRI to investigate the neural mechanisms that underlie adaptive goal-directed behavior in a cued task switching paradigm during real response conflict or, more generally, when expectations on the repetitive features of the environment were violated. Unexpected changes in sensory stimulus attributes in the currently unattended stimulus dimension thereby led to activations in a bilateral network comprising inferior lateral frontal, intraparietal, and posterior medial frontal brain regions, independent of whether these attributes elicited a factual response conflict or not. This fronto-parietal network may thus play an important role in adaptive responding to potentially significant events outside the current focus of attention. PMID- 19135770 TI - Long-term culture of primary human lymphoblastic leukemia cells in the absence of serum or hematopoietic growth factors. AB - OBJECTIVE: B-lineage acute lymphoblastic leukemia (ALL) and chronic myeloid leukemia in lymphatic blastic phase in adults have poor prognoses despite intensive chemotherapy. Novel targeted treatment modalities emerge, but their evaluation requires relevant in vitro models of lymphoblastic leukemia. Presently available cell lines do not fully represent this heterogeneous disease. Available in vitro culturing protocols do not support long-term proliferation of primary cells. We therefore aimed to develop a culture system that allows long-term proliferation of primary human B-lineage lymphoblastic leukemia. MATERIALS AND METHODS: Primary lymphoblastic leukemia cells were cultured in a defined serum free medium, in the absence or presence of human hematopoietic growth factors or serum. RESULTS: In the defined serum-free medium, cells from 12 of 34 cases immediately proliferated in vitro. In the absence of hematopoietic growth factors and serum these cases proliferated for more than 1 year without signs of exhaustion. The culturing system supported different subtypes of lymphoblastic leukemia. Two chronic myeloid leukemia in lymphatic blastic phase, four bcr/abl positive ALL, one etv6/abl-positive ALL, 2 e2a-pbx1-positive ALL, and one t(9;11) positive ALL could be long-term expanded, as well as two ALL that displayed nontypical cytogenetics. Not all bcr/abl- or e2a-pbx1-positive ALL proliferated in vitro, demonstrating heterogeneity within these subtypes. The proliferating bcr/abl- and etv6/abl-positive cells displayed sensitivity to imatinib, demonstrating that their proliferation depended on the activity of these oncoproteins. CONCLUSION: The serum-free culturing system may be a valuable instrument in the study of ALL cell biology, as well as in the evaluation of novel targeted therapeutics. PMID- 19135771 TI - Expression of P190 and P210 BCR/ABL1 in normal human CD34(+) cells induces similar gene expression profiles and results in a STAT5-dependent expansion of the erythroid lineage. AB - OBJECTIVE: The P190 and P210 BCR/ABL1 fusion genes are mainly associated with different types of hematologic malignancies, but it is presently unclear whether they are functionally different following expression in primitive human hematopoietic cells. MATERIALS AND METHODS: We investigated and systematically compared the effects of retroviral P190 BCR/ABL1 and P210 BCR/ABL1 expression on cell proliferation, differentiation, and global gene expression in human CD34(+) cells from cord blood. RESULTS: Expression of either P190 BCR/ABL1 or P210 BCR/ABL1 resulted in expansion of erythroid cells and stimulated erythropoietin independent burst-forming unit-erythroid colony formation. By using a lentiviral anti-signal transducer and activator of transcription 5 (STAT5) short-hairpin RNA, we found that both P190 BCR/ABL1- and P210 BCR/ABL1-induced erythroid cell expansion were STAT5-dependent. Under in vitro conditions favoring B-cell differentiation, neither P190 nor P210 BCR/ABL1-expressing cells formed detectable levels of CD19-positive cells. Gene expression profiling revealed that P190 BCR/ABL1 and P210 BCR/ABL1 induced almost identical gene expression profiles. CONCLUSIONS: Our data suggest that the early cellular and transcriptional effects of P190 BCR/ABL1 and P210 BCR/ABL1 expression are very similar when they are expressed in the same human progenitor cell population, and that STAT5 is an important regulator of BCR/ABL1-induced erythroid cell expansion. PMID- 19135772 TI - Optimization of methods for the detection of BCR-ABL activity in Philadelphia positive cells. AB - OBJECTIVE: The recent success in treating chronic myeloid leukemia (CML) with tyrosine kinase inhibitors (TKI), such as imatinib mesylate (IM), has created a demand for reproducible methods to accurately assess inhibition of BCR-ABL activity within CML cells, including rare stem and progenitor cells, either in vitro or in vivo. The purpose of this study was to develop an enzyme-linked immunosorbent (ELISA) method to measure total tyrosine phosphorylation (P-Tyr) in small samples of cells that express BCR-ABL and to compare to more established methods. MATERIALS AND METHODS: The assay was first validated in BCR-ABL wild type and mutant vs BCR-ABL-negative cell lines. P-Tyr levels were then measured by ELISA in primary CD34(+) CML cells treated with IM. RESULTS: In vitro exposure to TKI resulted in decreases in the level of P-Tyr, in both BCR-ABL-positive cell lines and primary CD34(+) CML samples, which were comparable to the reduction in P-Tyr by flow cytometry and phosphorylation of CrkL by either Western blot or flow cytometry. CONCLUSION: We have developed an accurate ELISA method to measure BCR-ABL activity within Ph(+) cells, which is comparable to other in vitro BCR ABL assessment techniques in terms of sensitivity and could be adapted for high throughput. PMID- 19135773 TI - Bone marrow mesenchymal stromal cells of patients with myeloproliferative disorders do not carry the JAK2-V617F mutation. AB - Myeloproliferative disorders (MPDs) are often associated with the presence of the JAK2-V617F mutation in hematopoietic cells. It is currently not known if this mutation is carried as well by bone marrow mesenchymal stromal cells (MSCs) in these patients. To test this hypothesis, we recruited seven patients with JAK2 V617F(+) MPD, isolated marrow MSCs and characterized their phenotype and mesenchymal differentiation capacity, and probed for JAK2-V617F genomic DNA mutation. We found that MSCs of most patients could be culture-expanded and had a phenotype and differentiation capacity similar to that of MSCs derived from normal subjects. Using real-time polymerase chain reaction and melting curve analysis with probes specific for the JAK2-V617F DNA mutation, we did not find the mutation in any of the MSC samples studied. These results demonstrate that, in the setting of MPD, MSC do not originate from the mutated hematopoietic progenitor clone. PMID- 19135774 TI - Interregional comparison of karst disturbance: west-central Florida and southeast Italy. AB - The karst disturbance index (KDI) consists of 31 environmental indicators contained within the five broad categories: geomorphology, hydrology, atmosphere, biota, and cultural. The purpose of this research is to apply the KDI to two distinct karst areas, west Florida, USA, and Apulia, Italy. Through its application, the utility of the index can be validated and other important comparisons can be made, such as differences in the karst legislations implemented in each region and the effect of time exposure to human occupation to each karst terrain. Humans have intensively impacted the karst of southeast Italy for thousands of years compared to only decades in west-central Florida. However, west-central Florida's higher population density allows the region to reach disturbance levels comparable to those reached over a longer period in Apulia. Similarly, Italian karst is more diverse than the karst found in west-central Florida, creating an opportunity to test all the KDI indicators. Overall, major disturbances for southeast Italy karst include quarrying, stone clearing, and the dumping of refuse into caves, while west-central Florida suffers most from the infilling of sinkholes, soil compaction, changes in the water table, and vegetation removal. The application of the KDI allows a benchmark of disturbance to be established and later revisited to determine the changing state of human impact for a region. The highlighting of certain indicators that recorded high levels of disturbance also allows regional planners to allocate resources in a more refined manner. PMID- 19135775 TI - Farm diversity, classification schemes and multifunctionality. AB - Diversity is not only intrinsic to agriculture; it can be considered also as one of its main assets as it provides a wide range of responses that can help to face uncertain futures. The ongoing encounter between changing spatial and temporal frameworks and a set of diverse farming strategies is leading to the emergence of an ongoing flow of development models that could materialize in a wide range of farming practices, contrasting enterprise models, changing relations between rural households and agricultural holdings, and differentiated patterns that link farming and farming families to the wider context in which they are embedded. The many-sided diversity encountered in agriculture is not only the outcome of the agency and polyvalence of the actors involved; their agency and polyvalence are in turn inspired and strengthened by the material and symbolic diversity, which contributes to a further unfolding of diversity. A proper understanding of the range, dimensions, significance and causes of diversity has been, over the centuries, a main concern--first for what is now known as classical agronomy, and later on in agrarian sciences. Yet the classification schemes, developed and used for such an understanding, have increasingly become an Achilles heel as each of them relies on specific assumptions that will bring out particular features of the overall farm diversity and will result in different perspectives of what agriculture is and how it fits into societal projects. Consequently, they are at the core of many debates and struggles, not only within agrarian sciences but increasingly on a wider societal level. The growing recognition of multifunctionality in agriculture, especially in the context of the changing EU policy, strengthens the relevance and importance of this debate. In this new context, we discuss advantages and limits of different classification principles by comparing two methodologies which have been extensively used in France and The Netherlands. PMID- 19135776 TI - Prognostic value of capsular invasion for localized clear-cell renal cell carcinoma. AB - BACKGROUND: The impact of capsular invasion on the survival of patients undergoing surgery for renal cell carcinoma (RCC) has attracted little attention in the literature and remains controversial. OBJECTIVES: To evaluate the value of capsular invasion, without perirenal fat invasion, on the prognosis of patients with localized clear-cell RCC. DESIGN, SETTING, AND PARTICIPANTS: Between 1984 and 2007, we retrospectively reviewed the records of 317 consecutive patients with localized clear-cell RCC (pT1-T2N0M0) who underwent radical nephrectomy or nephron-sparing surgery at our institution. Overall, 299 patients were eligible for the study. We analyzed clinical (presentation and body mass index [BMI]) and pathologic (tumor size, Fuhrman nuclear grade, collecting system invasion, microvascular invasion, and capsular involvement) parameters. MEASUREMENTS: Recurrence-free survival (RFS) and cancer-specific survival (CSS) were investigated using the Kaplan-Meier method, and the Cox regression model was used to determine the significant prognostic factors based on multivariate analysis. RESULTS AND LIMITATIONS: Renal capsular invasion was observed in 106 of 299 patients (35.5%). Capsular invasion had a statistically significant association with age, symptomatic presentation, tumor diameter, pathologic stage, collecting system invasion, and microvascular invasion. The mean follow-up was 60.5 mo (range: 1-249). The 5-yr RFS and CSS rates for tumors with capsular invasion were significantly lower compared with rates for tumors without invasion (77.7% vs 92.3% and 85.5% vs 95.7%, respectively; p=0.0004). Multivariate analysis showed that BMI (hazard ratio [HR]=0.19), stage (HR=2.45), and capsular invasion (HR=3.36) were independent prognostic factors of disease recurrence. With respect to CSS, BMI (HR=0.20), tumor size (HR=1.13), and capsular invasion (HR=4.03) were the factors related to death. Nevertheless, we recognize that these findings may be limited by the study's retrospective, single-institution design. CONCLUSIONS: Our findings suggest that capsular invasion is associated with poor survival in patients with localized clear-cell RCC. PMID- 19135777 TI - Editorial comment on: Prognostic value of capsular invasion for localized clear cell renal cell carcinoma. PMID- 19135778 TI - Down-regulation of c-Src/EGFR-mediated signaling activation is involved in the honokiol-induced cell cycle arrest and apoptosis in MDA-MB-231 human breast cancer cells. AB - Honokiol is a naturally occurring neolignan abundant in Magnoliae Cortex and has showed anti-proliferative and pro-apoptotic effects in a wide range of human cancer cells. However, the molecular mechanisms on the anti-proliferative activity in cancer cells have been poorly elucidated. In this study, we evaluated the growth inhibitory activity of honokiol in cultured estrogen receptor (ER) negative MDA-MB-231 human breast cancer cells. Honokiol exerted anti proliferative activity with the cell cycle arrest at the G0/G1 phase and sequential induction of apoptotic cell death in a concentration-dependent manner. The honokiol-induced cell cycle arrest was well correlated with the suppressive expression of CDK4, cyclin D1, CDK2, cyclin E, c-Myc, and phosphorylated retinoblastoma protein (pRb) at Ser780. Apoptosis caused by honokiol was also concomitant with the cleavage of caspases (caspase-3, -8, and -9) and Bid along with the suppressive expression of Bcl-2, but it was independent on the expression of Bax and p53. In addition, honokiol-treated cells exhibited the cleavage of poly (ADP-ribose) polymerase (PARP) and DNA fragmentation. In the analysis of signal transduction pathway, honokiol down-regulated the expression and phosphorylation of c-Src, epidermal growth factor receptor (EGFR), and Akt, and consequently led to the inactivation of mTOR and its downstream signal molecules including 4E-binding protein (4E-BP) and p70 S6 kinase. These findings suggest that honokiol-mediated inhibitory activity of cancer cell growth might be related with the cell cycle arrest and induction of apoptosis via modulating signal transduction pathways. PMID- 19135779 TI - Re-use of fluoride contaminated bone char sludge in concrete. AB - Managing sludge generated by treating groundwater contaminated with geogenic contaminants (fluoride, arsenic, and iron) is a major issue in developing nations. Their re-use in civil engineering applications is a possible pathway for reducing the impact on the geo-environment. This paper examines the re-use of one such sludge material, namely, fluoride contaminated bone char sludge, as partial replacement for fine aggregate (river-sand) in the manufacture of dense concrete specimens. Bone char sludge is being produced by defluoridation of contaminated groundwater in Nalagonda District, Andhra Pradesh, India. The impact of admixing 1.5-9% sludge contents on the compression strength and fluoride leaching potential of the sludge admixed concrete (SAC) specimens are examined. The compression strengths of the SAC specimens are examined with respect to strength criteria for manufacture of dense, load-bearing concrete blocks. The fluoride release potential of the SAC specimens is examined with respect to standards specific to disposal of treated leachate into inland surface water. PMID- 19135780 TI - Suitability of nitrogen rich compounds for gun propellant formulations. AB - This paper reports the suitability of a novel nitrogen rich compound, guanidinium 5-aminotetrazolate for RDX-based high-energy gun propellant formulations in respect of flame temperature as well as the burning rate characteristics. It has been found that the partial replacement of RDX with guanidinium-5-amino tetrazolate at the rate of five parts decreases the flame temperature of the propellant by about 120 K without adversely affecting the burning rate characteristics, i.e. linear rate of burning co-efficient and pressure exponent. PMID- 19135781 TI - Treatment of a synthetic solution of galvanization effluent via the conversion of sodium cyanide into an insoluble safe complex. AB - Wastewater discharged from metal-finishing processes usually contains cyanide, a hazardous substance that is used extensively in the surface finishing industry. In the present study, a synthetic solution resembling the contaminated wastewater was chemically treated using ferrous sulfate. This simple one-step process was applied successfully to remove cyanide from metal finishing wastewater. The experiments were carried out on a synthetic solution containing ions of cyanide and zinc. The effects of mixing velocity, ratio of ferrous ions to cyanide ions, ferrous ions concentration, initial cyanide concentration, pH of solution, temperature, mixing time and zinc ions concentration were studied. The results showed that the removal efficiency of cyanide increased as the mixing velocity increased, ferrous ion molar ratio to cyanide ions increased, temperature increased and time of mixing increased at an optimum of pH 8. The reduction of cyanide concentration reached the allowable limit for wastewater discharge according to the Egyptian Environmental Law decree 44/2000, which is 0.2mg/l. The formed complexes were analyzed and the stability of each complex was studied under different pH value solutions after 7 days of contact. A typical example of electroplating wastewater from a local company in the field of metal finishing, which contains 18 mg/lCN(-) and 12 mg/l Zn(2+), was treated according to the determined optimum conditions for the treatment process and the concentration of CN(-) was reduced to 0.095 mg/l after 15 min of agitation. PMID- 19135782 TI - Removal of some metal ions by activated carbon prepared from Phaseolus aureus hulls. AB - Removal of lead [Pb(II)], zinc [Zn(II)], copper [Cu(II)], and cadmium [Cd(II)] from aqueous solutions using activated carbon prepared from Phaseolus aureus hulls (ACPAH), an agricultural waste was studied. The influence of various parameters such as effect of pH, contact time, adsorbent dose, and initial concentration of metal ions on the removal was evaluated by batch method. The removal of metal ions by ACPAH was pH dependent and the optimum pH values were 7.0, 8.0, 7.0 and 6.0 for Cu(II), Cd(II), Zn(II), and Pb(II), respectively. The sorption isotherms were studied using Langmuir, Freundlich, Dubinin-Radushkevich (D-R), and Temkin isotherm models. The maximum adsorption capacity values of ACPAH for metal ions were 21.8 mg g(-1) for Pb(II), 21.2 mg g(-1) for Zn(II), 19.5 mg g(-1) for Cu(II), and 15.7 mg g(-1) for Cd(II). The experiments demonstrated that the removal of metal ions followed the pseudo-second-order kinetic model. Desorption experiments were carried out using HCl solution with a view to regenerate the spent adsorbent and to recover the adsorbed metal ions. PMID- 19135783 TI - Study of kinetic and fixed bed operation of removal of sulfate anions from an industrial wastewater by an anion exchange resin. AB - Sulfate anions represent very important wastewater pollutants, which appear in the effluents discharged from copper mines. In this study, for the first time, an attempt has been made on the removal of sulfate anions by an ion exchange resin. This work is focused on the removal of sulfate anions from the Sarcheshmeh copper complex (Kerman province, Southeast of Iran) wastewater by an anion exchange resin. Batch experiments of sulfate anions adsorption on Lewatit K6362 resin were carried out to determine the adsorption equilibrium data and the relation of adsorption isotherms. Isothermal data can be fitted with Freundlich adsorption isotherms better than Langmuir equation. The results show that maximum removal of sulfate anions take places in the resin dosage of 1000 mg/100ml and the adsorption of sulfate anions on the resin follows reversible first-order kinetics. The overall adsorption rate constants were compared for different initial concentrations. Finally, the effects of parameters such as the flow rate, bed height and inlet adsorbate concentration on the breakthrough curve in a fixed bed column were studied in detail. PMID- 19135784 TI - Separation of copper from aqueous sulfate solutions by mixtures of Cyanex 301 and LIX 984N. AB - The extraction equilibria of copper(II) with Cyanex 301, LIX 984N, and their mixtures have been investigated. Extraction was studied as a function of organic phase composition, sulfuric acid concentration, pH, temperature, initial copper concentration, mixing speed, and aqueous/organic volume ratio. Considerable synergistic enhancement has been observed in the extraction of Cu(2+) with mixtures of Cyanex 301 and LIX 984N. The results demonstrate that copper ion is extracted as CuRL(2)H with synergistic mixture. The thermodynamic parameter, enthalpy change (Delta H) of Cyanex 301, LIX 984N, and their mixtures have been determined and the endothermic process has been found. The synergistic enhancement factor of copper(II) with mixtures is higher at more acidic solutions, which suggests that it is a promising synergistic extraction system for the separation of copper(II) from more acidic medium. HCl was found to be more efficient for copper stripping from loaded synergistic mixtures. PMID- 19135785 TI - Oxidation by Fenton's reagent combined with biological treatment applied to a creosote-comtaminated soil. AB - In this study, we investigated the feasibility of using Fenton oxidation to remove sorbed polycyclic aromatic hydrocarbons (PAHs) in aged soil samples with creosote oil from a wood preserving site. The optimal dosage of reagents was determined by a statistical method, the central composite rotatable experimental design. The maximum PAH removal was 80% with a molar ratio of oxidant/catalyst equal to 90:1. In general low molecular weight PAHs (3 rings) were degraded more efficiently than higher molecular weight PAHs (4 and 5 rings). The hydrogen peroxide decomposition kinetic was studied in the presence of KH(2)PO(4) as stabilizer. The kinetic data were fitted to a simple model, the pseudo-first order which describes the hydrogen peroxide decomposition. The PAH kinetic degradation was also studied, and demonstrated that non-stabilized hydrogen peroxide was consumed in less than 30 min, whilst PAH removal continued for up to 24h. In a second part of the work, a combined chemical and biological treatment of the soil was carried out and shown to be dependent on the pre-oxidation step. Different reagent doses (H(2)O(2):Fe) were used (10, 20, 40, 60:1) in the pre treatment step. An excess of hydrogen peroxide resulted in a poor biological removal, thus the optimal molar ratio of H(2)O(2):Fe for the combined process was 20:1. The combined treatment resulted in a maximum total PAH removal of 75% with a 30% increase in removal due to the biodegradation step. The sample with highest PAH removal in the pre-oxidation step led to no further increase in removal by biological treatment. This suggests that the more aggressive chemical pre oxidation does not favour biological treatment. The physico-chemical properties of the pollutants were an important factor in the PAH removal as they influenced chemical, biological and combined treatments. PMID- 19135786 TI - Rapid removal of cobalt ion from aqueous solutions by almond green hull. AB - Almond green hull, an agriculture solid waste, was chemically treated and used for the adsorption of Co (II) from aqueous solutions. The efficiency of this new adsorbent was studied using batch adsorption technique under different experimental conditions such as sorbent amount, initial metal-ion concentration, contact time, adsorbent particle size, and chemical treatment. Optimum dose of sorbent for maximum metal-ion adsorption were 0.25 g for 51.5 mg l(-1) and 0.4 g for 110 mg l(-1) solutions, respectively. High removal efficiencies of Co (II) were occurred in the first 1 min of sorbent contact time. The adsorption of Co (II) on almond green hull was also observed to follow the pseudo second-order kinetics. Adsorption isotherms were expressed by Langmuir and Freundlich adsorption models. The Langmuir adsorption model fits the experimental data reasonably well compared to the Freundlich model. The maximum adsorption capacity of this new sorbent was found to be 45.5 mg g(-1). The present study revealed that such a low cost material could be used as an efficient sorbent for the removal of cobalt from wastewater streams. PMID- 19135787 TI - Reactions of sulphur mustard and sarin on V 1.02 O 2.98 nanotubes. AB - Reactions of sulphur mustard and sarin were studied on the surface of V(1.02)O(2.98) nanotubes by gas chromatography and gas chromatography-mass spectrometry techniques. The V(1.02)O(2.98) nanotube samples were made by using hydrothermal method and characterized by scanning electron microscopy, nitrogen adsorption, X-ray diffractometry and thermogravimetry. Later, they were exposed to sulphur mustard and sarin separately at ambient temperature (30+/-2 degrees C). The data explored the formation of sulphoxide of sulphur mustard, thiodiglycol for sulphur mustard and isopropyl methyl phosphonic acid for sarin on V(1.02)O(2.98) nanotubes illustrating the role of oxidation and hydrolysis reactions in the decontamination. PMID- 19135788 TI - Synthesis of high capacity cation exchangers from a low-grade Chinese natural zeolite. AB - The Chinese natural zeolite, in which clinoptilolite coexists with quartz was treated hydrothermally with NaOH solutions, either with or without fusion with NaOH powder as pretreatment. Zeolite Na-P, Na-Y and analcime were identified as the reacted products, depending on the reaction conditions such as NaOH concentration, reaction time and hydrothermal temperature. The products were identified by X-ray diffraction, and characterized by Fourier transform IR and ICP. With hydrothermal treatment after fusion of natural zeolite with NaOH, high purity of zeolite Na-Y and Na-P can be selectively formed, their cation exchange capacity (CEC) are 275 and 355 meq/100g respectively, which are greatly higher than that of the natural zeolite (97 meq/100g). Furthermore, the ammonium removal by the synthetic zeolite Na-P in aqueous solution was also studied. The equilibrium isotherms have been got and the influence of other cations present in water upon the ammonia uptake suggested an order of preference Ca(2+)>K(+)>Mg(2+). PMID- 19135789 TI - Simple correlation for predicting detonation velocity of ideal and non-ideal explosives. AB - This paper describes a simple method for prediction of detonation velocity of ideal and non-ideal explosives. A non-ideal aluminized and nitrated explosive can have Chapman-Jouguet detonation velocity significantly different from that expected from existing thermodynamic computer codes for equilibrium and steady state calculations. Detonation velocity of explosives with general formula C(a)H(b)N(c)O(d)Al(e) can be predicted only from values of a, b, c, d, e and a specific structural parameter without using any assumed detonation products, heat of formation and experimental data. Predicted detonation velocities by this procedure for ideal and non-ideal explosives show good agreement with respect to experimental values as compared to computed results of BKWR and BKWS equations of state. PMID- 19135790 TI - Catalytic wet peroxide oxidation of azo dye (Congo red) using modified Y zeolite as catalyst. AB - The present study explores the degradation of azo dye (Congo red) by catalytic wet peroxide oxidation using Fe exchanged commercial Y zeolite as a catalyst. The effects of various operating parameters like temperature, initial pH, hydrogen peroxide concentration and catalyst loading on the removal of dye, color and COD from an aqueous solution were studied at atmospheric pressure. The percent removals of dye, color and COD at optimum pH(0) 7, 90 degrees C using 0.6 ml H(2)O(2)/350 ml solution and 1g/l catalyst was 97% (in 4h), 100% (in 45 min) and 58% (in 4h), respectively. The % dye removal has been found to be less in comparison to % color removal at all conditions, e.g. dye removal in 45 min and at above conditions was 82%, whereas the color removal was 100%. The results indicate that the Fe exchanged Y zeolite is a promising catalyst for dye removal. Fe exchanged catalyst is characterized using XRD, SEM/EDAX, surface area analyzer and FTIR. Though the dye, color and COD removals were maximum at pH(0) 2 but as the leaching of Fe from the catalyst was more in acidic pH range, pH(0) 7 was taken as operating pH due to almost comparable removals as of pH(0) 2 and no leaching of Fe ions. PMID- 19135791 TI - Pretreatment of ammonium removal from landfill leachate by chemical precipitation. AB - Chemical precipitation is a useful technology to remove ammonium (NH(4)(+)) from landfill leachate. In this paper, the removal of ammonium from landfill leachate was studied. The objective of this study was to investigate optimum pH, optimum molar ratio, and different kinds of chemicals combinations for magnesium ammonium phosphate (MAP) precipitation. Furthermore, the kinetics of MAP formation and surface characterization were analyzed. The results indicated that ammonium in landfill leachate could be removed with the optimum pH of 9.5. The Mg(2+):NH(4)(+):PO(4)(3-) molar ratio was practically controlled at 1.15:1:1 to remove ammonium effectively and avoid higher concentration of PO(4)(3-) in the effluent. Highest salt concentration was generated by using MgCl(2).6H(2)O plus Na(2)HPO(4).12H(2)O. Compare to MgCl(2).6H(2)O and Na(2)HPO(4).12H(2)O, adding MgO and 85% H(3)PO(4) could significantly minimize the salt concentration, although ammonium removal ratio was 9 percent lower. The lowest ammonium removal ratio was generated by adding Ca(H(2)PO(4))(2).H(2)O and MgSO(4).7H(2)O. Moreover, the kinetics experiment shown that the rate of reaction was closer to the first-order kinetic model. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis indicated that MAP was the main composition of the precipitates. Scanning electron microscopy with energy dispersive X-ray (SEM EDX) analysis indicated that the unshaped crystal was coarse and its size was irregular, the surface composition of the precipitates contains a great deal of O, P, Mg and trace of C, K, Na, Cl. PMID- 19135792 TI - Removal of oxyanions from synthetic wastewater via carbonation process of calcium hydroxide: applied and fundamental aspects. AB - Removal of oxyanions (selenite, selenate, arsenate, phosphate and nitrate) during calcite formation was experimentally studied using aqueous carbonation of calcium hydroxide under moderate pressure (P(CO2) congruent with 20 bar) and temperature (30 degrees C). The effects of Ca(OH)(2) dose (10 and 20 g), Ca(OH)(2) source (commercial pure material or alkaline paper mill waste) and oxyanion initial concentration (from 0 to 70 mg atom/L) were investigated for this anisobaric gas liquid-solid system. The Ca(OH)(2) carbonation reaction allowed successfully the removal of selenite (>90%), arsenate (>78%) and phosphate (congruent with 100%) from synthetic solutions. Conversely, nitrate and selenate had not any physicochemical affinity/effect during calcite formation. The rate of CO(2) transfer during calcite formation in presence of oxyanions was equal or slower than for an oxyanion-free system, allowing to define a retarding kinetic factor RF that can vary between 0 (no retarding effect) to 1 (total inhibition). For selenite and phosphate RF was quite high, close to 0.3. A small retarding effect was detected for arsenate (RF approximately 0.05) and no retarding effect was detected for selenate and nitrate (RF approximately 0). In general, RF depends on the oxyanion initial concentration, oxyanion nature and Ca(OH)(2) dose. The presence of oxyanions could also influence the crystal morphology and aggregation/agglomeration process. For example, a c-axis elongation of calcite crystals was clearly observed at the equilibrium, for calcite formation in presence of selenite and phosphate. The oxyanions removal process proposed herein was inspired on the common physicochemical treatment of wastewater using calcium hydroxide (Ca(OH)(2)). The particularity, for this novel method is the simultaneous calcium hydroxide carbonation with compressed carbon dioxide in order to stabilise the solid matter. This economical and ecological method could allow the removal of various oxyanions as well as the ex situ mineral sequestration of CO(2); particularly, when the Ca(OH)(2) source comes from alkaline solid waste. PMID- 19135793 TI - Synthesis of nano B2O3/TiO2 composite material as a new solid phase extractor and its application to preconcentration and separation of cadmium. AB - A new solid phase extractor, nano-scale diboron trioxide/titanium dioxide composite material, was synthesized and used for separation and/or preconcentration of trace cadmium ion from various samples. The characterization of the synthesized material was performed by scanning electron microscope (SEM), transmission electron microscope (TEM) and X-ray diffractometer methods (XRD). The specific surface area of the material was also determined and found as 3.4m(2)/g. Analytical parameters including pH of sample solution, sample volume, flow rate of sample solution, volume and concentration of eluent for the column solid phase extraction (SPE) procedure were examined. The effect of common matrix ions on the recovery of the cadmium has also been investigated and found that they did not interfere on cadmium preconcentration. Under the optimum experimental conditions, preconcentration factor and analytical detection limit were determined as 50 and 1.44 microg/L, respectively. The reusability (stabile up to 100 run) and adsorption capacity (49 mg/g) of the sorbent were excellent. The accuracy of the method was confirmed by analyzing certified reference materials (Tea leaves GBW-07605). The results demonstrated good agreement with the certified values (relative error <10%). The precision of the method was also satisfactory. The recovery of cadmium under the optimum conditions was found to be 96+/-3% at 95% confidence level. The method was applied for the determination of cadmium in tap water and tea leaves. PMID- 19135794 TI - Adsorption of organic compounds from aqueous solution onto the synthesized zeolite. AB - A novel zeolite was synthesized, characterized and employed for the adsorption of methylene blue (cationic dye) and bisphenol-A in aqueous solution. The pore properties of the synthesized zeolite have been determined using N(2) adsorption desorption isotherms, indicating that it is a supermicroporous adsorbent with BET surface area of over 400 m(2)g(-1). Based on the XRD image, it was indicative of the probable formation of zeolite-P2 in the hydrothermal synthesis. The metal content and zeta-potential of the zeolite were also measured to examine the hydrophilicity and the effect of pH on the surface charge, respectively. It was found that the synthesized zeolite exhibited significantly higher adsorption capacity for methylene blue than that for bisphenol-A due to the difference in molecular properties. Kinetic studies at 25 degrees C indicated that the adsorption of methylene blue well followed the pseudo-second-order model and could be elucidated by considering the pore property and surface charge of the synthesized zeolite. The kinetic parameters thus obtained from the fittings of the model were dependent on initial dye concentration, pH, and adsorbent mass. PMID- 19135795 TI - Characterization and activity studies of highly heavy metal resistant sulphate reducing bacteria to be used in acid mine drainage decontamination. AB - Biological treatment with sulphate-reducing bacteria (SRB) has been considered as the most promising alternative for acid mine drainage (AMD) decontamination. Normally, these wastewaters contain high concentrations of sulphate and heavy metals, so the search for SRB highly resistant to metals is extremely important for the development of a bioremediation technology. A SRB consortium resistant to high concentrations of heavy metals (Fe, Cu and Zn), similar to those typically present in AMD, was obtained among several environmental samples, from a wastewater treatment plant. The phylogenetic analysis of the dsr gene sequence revealed that this consortium contains species of SRB affiliated to Desulfovibrio desulfuricans and Desulfobulbus rhabdoformis. The results show that the presence of usually lethal concentrations of Fe (400mg/L), Zn (150 mg/L) and Cu (80 mg/L) is not toxic for the sulphate-reducing bacteria present in this sample. As a consequence, a very good efficiency in terms of sulphate reduction and metals removal was obtained. Both ethanol and lactate can be used by this inoculum as carbon source. With the other samples tested sulphate reduction was inhibited by the presence of copper and zinc. This highly metal resistant consortium will be used to inoculate a bioreactor to carry out AMD decontamination. PMID- 19135796 TI - Montmorillonite-supported magnetite nanoparticles for the removal of hexavalent chromium [Cr(VI)] from aqueous solutions. AB - Montmorillonite-supported magnetite nanoparticles were prepared by co precipitation and hydrosol method. The obtained materials were characterized by X ray diffraction, nitrogen adsorption, elemental analysis, differential scanning calorimetry, transmission electron microscopy and X-ray photoelectron spectroscopy. The average sizes of the magnetite nanoparticles without and with montmorillonite support are around 25 and 15 nm, respectively. The montmorillonite-supported magnetite nanoparticles exist on the surface or inside the interparticle pores of clays, with better dispersing and less coaggregation than the ones without montmorillonite support. Batch tests were carried out to investigate the removal mechanism of hexavalent chromium [Cr(VI)] by these synthesized magnetite nanoparticles. The Cr(VI) uptake was mainly governed by a physico-chemical process, which included an electrostatic attraction followed by a redox process in which Cr(VI) was reduced into trivalent chromium. The adsorption of Cr(VI) was highly pH-dependent and the kinetics of the adsorption followed the Pseudo-second-order model. The adsorption data of unsupported and clay-supported magnetite nanoparticles fit well with the Langmuir and Freundlich isotherm equations. The montmorillonite-supported magnetite nanoparticles showed a much better adsorption capacity per unit mass of magnetite (15.3mg/g) than unsupported magnetite (10.6 mg/g), and were more thermally stable than their unsupported counterparts. These fundamental results demonstrate that the montmorillonite-supported magnetite nanoparticles are readily prepared, enabling promising applications for the removal of Cr(VI) from aqueous solution. PMID- 19135797 TI - Repeated dosing of ABT-102, a potent and selective TRPV1 antagonist, enhances TRPV1-mediated analgesic activity in rodents, but attenuates antagonist-induced hyperthermia. AB - Transient receptor potential vanilloid type 1 (TRPV1) is a ligand-gated ion channel that functions as an integrator of multiple pain stimuli including heat, acid, capsaicin and a variety of putative endogenous lipid ligands. TRPV1 antagonists have been shown to decrease inflammatory pain in animal models and to produce limited hyperthermia at analgesic doses. Here, we report that ABT-102, which is a potent and selective TRPV1 antagonist, is effective in blocking nociception in rodent models of inflammatory, post-operative, osteoarthritic, and bone cancer pain. ABT-102 decreased both spontaneous pain behaviors and those evoked by thermal and mechanical stimuli in these models. Moreover, we have found that repeated administration of ABT-102 for 5-12 days increased its analgesic activity in models of post-operative, osteoarthritic, and bone cancer pain without an associated accumulation of ABT-102 concentration in plasma or brain. Similar effects were also observed with a structurally distinct TRPV1 antagonist, A-993610. Although a single dose of ABT-102 produced a self-limiting increase in core body temperature that remained in the normal range, the hyperthermic effects of ABT-102 effectively tolerated following twice-daily dosing for 2 days. Therefore, the present data demonstrate that, following repeated administration, the analgesic activity of TRPV1 receptor antagonists is enhanced, while the associated hyperthermic effects are attenuated. The analgesic efficacy of ABT-102 supports its advancement into clinical studies. PMID- 19135798 TI - No evidence for the development of acute tolerance to analgesic, respiratory depressant and sedative opioid effects in humans. AB - It is widely accepted that chronic opioid therapy is associated with the development of pharmacological tolerance. More controversial is the question as to whether acute opioid administration can result in "acute tolerance." The aim of this double-blind, placebo-controlled study in thirty-six healthy human volunteers was to examine whether a 3-h intravenous infusion delivering two different but clinically relevant doses of the mu-opioid receptor agonist remifentanil would result in tolerance to analgesic, respiratory depressant and/or sedative opioid effects. The blood remifentanil concentration versus opioid effect relationship was determined before and after the 3-h infusion. Tolerance was inferred if the potency of remifentanil was significantly lower after the 3-h infusion. Opioid analgesia was assessed with the aid of the cold pressor test and models of electrical and heat pain. Respiratory depression was assessed by measuring arterial pCO2 and minute ventilation. Subjective sedation scores were assessed on a visual analogue scale. Mixed effects modeling was used to relate the steady-state blood remifentanil concentration to each pharmacodynamic assessment. Neither dose of remifentanil produced detectable tolerance to any of the measured opioid effects following a 3-h infusion. The study was adequately powered to detect a decrease in potency of 5-24% for analgesia, 20-48% for respiratory depression, and 32% for sedative effects. These results suggest that short-term administration of clinically useful doses of remifentanil is not associated with the development of significant tolerance to analgesic, respiratory depressant, or sedative opioid effects. PMID- 19135799 TI - Acute, localised paroxysmal pain as the initial manifestation of focal seizures: a case report and a brief review of the literature. AB - Pain is a rare manifestation of epileptic seizures. Yet, despite its rarity as a clinical entity, the pain associated with such seizures can be both severe and disabling. It thus remains important to consider epilepsy in the differential diagnosis of unexplained paroxysmal pain. By way of illustration, we present here a report of acute localised paroxysmal pain as the initial manifestation of focal epilepsy with EEG abnormalities in an otherwise neurodevelopmentally normal child. A brief discussion of the literature on epileptic pain then follows. PMID- 19135801 TI - Import of Parafilaria bovicola in the Netherlands. AB - In March 2007 cutaneous nodules were observed in the neck-, shoulder- and back region of a breeding bull, imported from France in November 2006. The nodules opened spontaneously and produced a haemorrhagic exudate. After suspicion of parafilariosis, the bull was treated with moxidectin, but without effect. Treatment with ivermectin stopped the bleedings. To prevent spread of this imported infection, the bull was euthanised and necropsied. Worms were found in the subcutis which could be identified as Parafilaria bovicola. This is the first diagnosed case of parafilariosis in the Netherlands. In the following period, all cattle on the farm were carefully examined for signs of parafilariosis and provided with permethrin containing ear tags. Till July 2008, no new cases on the farm were observed. PMID- 19135800 TI - Periganglionic inflammation elicits a distally radiating pain hypersensitivity by promoting COX-2 induction in the dorsal root ganglion. AB - We have developed a model in which inflammation contiguous to and within a dorsal root ganglion (DRG) was generated by local application of complete Freund's adjuvant (CFA) to the L4 lumbar spinal nerve as it exits from the intervertebral foramen. The periganglionic inflammation (PGI) elicited a marked reduction in withdrawal threshold to mechanical stimuli and an increase in heat pain sensitivity in the ipsilateral hindpaw in the absence of any hindpaw inflammation. The pain sensitivity appeared within hours and lasted for a week. The PGI also induced a prominent increase in IL-1beta and TNF-alpha levels in the DRG and of cyclooxygenase-2 (COX-2) expression in neurons and satellite cells. A selective COX-2 inhibitor reduced the PGI-induced hyperalgesia. We also show that IL-1beta induces COX-2 expression and prostaglandin release in DRG neurons in vitro in a MAP kinase-dependent fashion. The COX-2 induction was prevented by ERK and p38 inhibitors. We conclude that periganglionic inflammation increases cytokine levels, including IL-1beta, leading to the transcription of COX-2 and prostaglandin production in the affected DRG, and thereby to the development of a dermatomally distributed pain hypersensitivity. PMID- 19135802 TI - Geometric means provide a biased efficacy result when conducting a faecal egg count reduction test (FECRT). AB - The process of conducting a faecal egg count reduction test was simulated to examine whether arithmetic or geometric means offer the best estimate of efficacy in a situation where the true efficacy is known. Two components of sample variation were simulated: selecting hosts from the general population which was modelled by the negative binomial distribution (NBD), and taking an aliquot of faeces from the selected host to estimate the worm egg count by assuming a Poisson distribution of sample counts. Geometric mean counts were determined by adding a constant (C) to each count prior to log transformation, C was set at 25, 12 or 1. Ten thousand Monte Carlo simulations were run to estimate mean efficacy, the 2.5% (lower) and the 97.5% (upper) percentile based on arithmetic or geometric means. Arithmetic means best estimated efficacy for all different levels of worm aggregation. For moderate levels of aggregation and with C=1 the geometric mean substantially overestimated efficacy. The bias was reduced if C was increased to 25 but the results were no better than those based on arithmetic means. For very high levels of aggregation (over-dispersed populations) the geometric mean underestimated efficacy regardless of the size of C. It is recommended that the guidelines on anthelmintic resistance be revised to advocate the use of arithmetic means to estimate efficacy. PMID- 19135803 TI - In vitro effects of Cucurbita moschata seed extracts on Haemonchus contortus. AB - This study was carried out to evaluate the in vitro effect of Cucurbita moschata seed against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and dichloromethane) of C. moschata seed were tested in vitro on four developmental stages of H. contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition (LMI) assay and adult worm motility (AWM) assay. The highly significant (P<0.001) ability to stop larval development (inhibition>90% for each extract) and the negative effect of the dichloromethane and methanolic extracts on adult worm motility (inhibition of motility >59.2% after 24h of incubation) compared to the negative controls, suggest anthelmintic properties of C. moschata seed against H. contortus. The active principles responsible for the activity could be secondary metabolites such as amino acid compounds or terpenoid compounds present in the extracts. PMID- 19135804 TI - Prevalence of Calodium hepaticum (Syn. Capillaria hepatica) in house mice (Mus musculus) in the Azores archipelago. AB - Calodium hepaticum (Syn. Capillaria hepatica) is a zoonotic liver nematode of mammals distributed worldwide. Rodents are believed to be the main reservoirs of this nematode. In this paper, prevalence of the parasite was analyzed in liver histological sections from 51 house mice (Mus musculus) caught in human-inhabited houses, from two localities (Furnas and Rabo de Peixe) on Sao Miguel island from the Azores archipelago (Portugal). Mean prevalence of infection was 19.6%, with 33.3% prevalence in Furnas and 4.1% in Rabo de Peixe (P=0.07). No significant differences were found between the prevalence of infection and the age, body weight and the sex of mice. Hepatic lesions found were either acute and/or chronic stage and consisted of moderate to severe multifocal pyogranulomatous hepatitis with encapsulated eggs with typical bipolar plugs and moderate to severe necrotizing hepatitis consistent with larva tracks. Periportal inflammatory infiltration, hepatocyte regeneration and bile duct hyperplasia were also noted. In most cases, hepatic lesions occupied more than 50% of the liver, but despite severe lesions, in some mice, no signs of hepatic failure were noticed. The high rate of infection found in the present study suggests that house mice are an important reservoir for this parasite in the Azores and could have a role in human transmission. PMID- 19135806 TI - Pretreatment with alternate day modified fast will permit higher dose and frequency of cancer chemotherapy and better cure rates. AB - It is established that calorie restriction (CR) increases the resistance of cells to various stressors such as oxidative damage, excitotoxins, mercury and acetaminophen. Alternate day feeding (ADF) may confer greater stress resistance than daily CR of 30% or 40%. A recent study in three strains of mouse showed that a fast of 48 or 60 h prevented toxic effects due to administration of doses 2-4 times the maximum human dose of etoposide, a chemotherapy agent which acts through increased oxidative stress. In addition, mice inoculated with neuroblastoma survived longer when pretreated with fasting, then given high dose etoposide, as well as not exhibiting toxicity. This increased survival was construed as evidence of differential stress resistance between normal and cancer cells, the cancer cells being only partially protected by the pretreatment fast. In clinical practice, increased differential stress resistance could lead to the use of much higher doses of chemotherapy agents, and in the absence of toxicity, make it possible to repeat the treatment to kill residual cancer cells. Humans are unlikely to comply with a total fast of longer than 24 or 48 h, which may be insufficient to activate the same gene expression process. Based on published data we estimate that an optimal time period for development of stress resistance is 2-3 weeks when alternate day feeding is employed. Our previously published experience suggests that 2-3 weeks of alternate day modified fast in which subjects eat ad libitum one day and <20% of one's estimated caloric requirement the next will confer a similar stress resistance. Compliance with this diet is high and greater maintenance of body weight is feasible. We hypothesize that a pretreatment of 2-3 weeks with the alternate day modified fast will improve outcomes in cancer chemotherapy, decreasing morbidity and raising cure rates. PMID- 19135805 TI - Diurnal cortisol amplitude and fronto-limbic activity in response to stressful stimuli. AB - The development and exacerbation of many psychiatric and neurologic conditions are associated with dysregulation of the hypothalamic pituitary adrenal (HPA) axis as measured by aberrant levels of cortisol secretion. Here we report on the relationship between the amplitude of diurnal cortisol secretion, measured across 3 typical days in 18 healthy individuals, and blood oxygen level dependant (BOLD) response in limbic fear/stress circuits, elicited by in-scanner presentation of emotionally negative stimuli, specifically, images of the World Trade Center (WTC) attack. Results indicate that subjects who secrete a greater amplitude of cortisol diurnally demonstrate less brain activation in limbic regions, including the amygdala and hippocampus/parahippocampus, and hypothalamus during exposure to traumatic WTC-related images. Such initial findings can begin to link our understanding, in humans, of the relationship between the diurnal amplitude of a hormone integral to the stress response, and those neuroanatomical regions that are implicated as both modulating and being modulated by that response. PMID- 19135807 TI - Carbonic anhydrase III: the new hope for the elimination of exercise-induced muscle fatigue. AB - Fatigue, defined as the failure to maintain the required or expected power output, is a complex problem. Its occurrence mechanism is extremely complicated. The obvious reasons are that it is a multifactorial situation and that the limiting factors may vary with force intensity, exercise duration and muscle type. In recent years, it has been found that carbonic anhydrase III(CAIII) which is present in high concentrations in muscles has multiple biological activities that can dissipate or resist some fatigue related substances. Therefore, we hypothesize that the CAIII supplementation may contribute to dissipate fatigue. Confirmation of this hypothesis will further add to our understanding of the physiological functions of CAIII and will be hopeful to solve the difficulties in eliminating muscular fatigue. PMID- 19135808 TI - Surgical treatment of multifocal giant cell tumor of carpal bones with preservation of wrist function: case report. AB - We report a rare case of multifocal giant cell tumor of bone involving the trapezium, trapezoid, capitate, and scaphoid with soft tissue extension. Following intralesional resection, an autogenous corticocancellous iliac crest bone graft was used to fill the resultant defect and preserve carpal height and radiocarpal motion. Successful union with no recurrence was noted at 1-year follow-up. PMID- 19135809 TI - Arthroscopic findings in gout of the wrist. AB - PURPOSE: We have recently treated several patients with gout of the wrist and scapholunate ligament pathology. The purpose of this investigation was to report the arthroscopic findings of 7 such patients. METHODS: We searched our medical records for patients with a diagnosis of gout and nontraumatic wrist pathology. We retrospectively reviewed the clinical, arthroscopic, radiographic, and pathologic findings in 7 patients who failed nonsurgical management and were treated with wrist arthroscopy. RESULTS: All 7 wrists had diffuse synovitis and crystalline deposits throughout the radiocarpal joint. There were focal crystalline precipitates on the scapholunate and lunotriquetral ligaments but not on the triangular fibrocartilage complex. Scapholunate or lunotriquetral ligament disruption was noted in 6 and 5 wrists, respectively. An early scapholunate advanced collapse pattern of arthritis was present in 5 patients. CONCLUSIONS: Gout affecting the wrist may be associated with crystal deposition on the scapholunate and lunotriquetral ligaments. Ligament incompetence and a scapholunate advanced collapse pattern of wrist arthritis was noted in 6 of our 7 patients. PMID- 19135810 TI - TNF-alpha antagonists and other recombinant proteins for the treatment of rheumatoid arthritis. PMID- 19135811 TI - Molecular cloning, sequencing and expression of the outer membrane protein A gene from Haemophilus parasuis. PMID- 19135812 TI - Analysis of the occurrence and distribution of the Omp25/Omp31 family of surface proteins in the six classical Brucella species. AB - Members of the Omp25/Omp31 family of surface proteins were previously shown to participate in the virulence of some Brucella species and a different distribution of the seven proteins of this family among species could be related to the difference in pathogenicity and host preference they exhibit. Accordingly, in this work we have analyzed the expression of the genes coding for the Omp25/Omp31 family in the six classical Brucella species and a set of B. ovis mutant strains with each omp gene inactivated. Immunoblot of whole-cell lysates with antibodies raised against the purified recombinant outer membrane proteins (OMPs) did not show the simultaneous presence of the seven OMPs in any of the Brucella strains studied, but different Omp25/Omp31 profiles were detected, in our experimental conditions, between the Brucella strains representative of the six classical species. Transcripts for omp31, omp25 and omp25c were, in general, the most abundant of the family and some hits were found in B. ovis for a posttranscriptional regulation mechanism and for a compensatory mechanism increasing the synthesis of a protein to compensate for the absence of another one. Finally, the potential interest of Omp25c and Omp31b as subcellular vaccines, considering their occurrence in the Brucella strains studied and their antigenic relatedness with other proteins of the family, is discussed. PMID- 19135813 TI - Pathogenesis of Mycobacterium avium subsp. paratuberculosis in neonatal calves after oral or intraperitoneal experimental infection. AB - Understanding the host response to Mycobacterium avium subsp. paratuberculosis is critical to the development of effective vaccines and therapeutics for the control of this disease in the field. The current study compared the effectiveness of oral and intraperitoneal (IP) methods of experimental inoculation and two strains of M. avium subsp. paratuberculosis (strain K-10 and clinical isolate 509) on the level of infection and lesion development. Calves were inoculated with 4x10(11) to 8x10(12)cfu live bacteria, depending upon treatment group. Fecal shedding of M. avium subsp. paratuberculosis was minimal and infrequent over the course of the study for calves that received strain K-10 (oral and IP), however, calves orally inoculated with the clinical isolate shed high numbers of bacteria in their feces up to 4 months post-inoculation. Colonization was present in a number of intestinal tissues and lymph nodes with the lowest number of affected tissues in the IP calves and the highest for calves receiving the clinical isolate via oral inoculation. Microscopic lesions were predominantly found in the ileal and jejunal sections of small intestine and their associated lymph nodes, as well as the ileocecal valve and node. These data suggest that a variety of experimental infection regimes can be effective but oral inoculation with a clinical isolate may result in greater colonization of tissues and fecal shedding of M. avium subsp. paratuberculosis. PMID- 19135814 TI - Brain imaging findings in very preterm infants throughout the neonatal period: part II. Relation with perinatal clinical data. AB - This study describes the relation between frequent and clinically relevant brain imaging findings in very preterm infants (GA<32 weeks), assessed with sequential cranial ultrasonography throughout the neonatal period and MRI around term age, and several potential perinatal risk factors. For ultrasound findings during admission the following independent risk factors were identified: male gender for periventricular echodensities and intraventricular haemorrhage, postnatal corticosteroid treatment for cystic white matter lesions, and lower gestational age for post-haemorrhagic ventricular dilatation. For MRI findings around term age, including punctate white matter lesions, ventricular dilatation, decreased cortical complexity, and diffuse and excessive high signal intensity, no independent risk factors were found. In very preterm infants, the risk factors for frequently found changes on cranial ultrasound have largely remained unchanged over the last decades, while no risk factors could be identified for subtle and diffuse white matter injury as seen on MRI around term age. PMID- 19135816 TI - Impact of image noise levels, scout scan dose and lens shield on image quality and radiation exposure in z-axis dose-modulated neck MSCT on 16- and 64-slice Toshiba Aquilion scanners. AB - OBJECTIVE: Assessing the impact of image noise (IN) levels, scout scan dose and lens shield use on image quality and radiation exposure in neck multislice CT (MSCT) when using z-axis dose modulation (DM). METHODS: Neck MSCT phantom studies with/without z-axis DM were performed by using different IN levels (S.D. 7.5 30HU) and scout scan tube currents (7.5-50mA) on Toshiba Aquilion scanners (16 /64-slice). Image quality indices were evaluated by two radiologists and radiation exposure parameters calculated. Cadaveric phantom measurements elucidated lens shield interactions with DM efficacy. The lowest dose scan protocol with diagnostic image quality was introduced into the clinical imaging routine and retrospectively evaluated in 20 age-matched patients undergoing neck MSCT with/without DM. RESULTS: The highest image noise level in DM neck studies with comparable image quality to standard neck CT amounted to 20HU, resulting in a mean tube current of 50mAs (CTDI(w) 6.3mGy). DM reduced effective dose by 35% and organ dose figures (lens, thyroid) by 33%. Scout scan dose lowering to 20mA resulted in an effective dose (ED) decrease of 0.06mSv (5%). Avoiding lens shield placement during scout scan effected an organ dose decrease of 20%. Overall contour sharpness and image contrast did not differ significantly (DM/without DM) whereas image noise was rated higher in DM neck CT studies (p<0.05). CONCLUSIONS: z-Axis dose modulation, as assessed on 16- and 64-slice Toshiba Aquilion scanners, is effective and mandatory in neck MSCT. DM efficacy can be enhanced by optimising scout scan doses and lens shield use. PMID- 19135815 TI - Use of fission yeast heterologously expressing human cytochrome P450 2B6 in biotechnological synthesis of the designer drug metabolite N-(1-phenylcyclohexyl) 2-hydroxyethanamine. AB - Standards of drug metabolites are required for drug metabolism studies as a basis for toxicological risk assessment with respect to drug interactions and pharmacogenetic polymorphisms. They are further needed as reference compounds in analytical toxicology. However, metabolite standards are often not commercially available, particularly in the case of new designer drugs. As an alternative to often cumbersome chemical synthesis, human cytochrome P450 (CYP) isoenzymes heterologously expressed in the fission yeast Schizosaccharomyces pombe can be used for the biotechnological synthesis of drug metabolites. In the present study this concept was applied to the synthesis of N-(1-phenylcyclohexyl)-2 hydroxyethanamine (PCHEA), the common O-dealkyl metabolite of the designer drugs N-(1-phenylcyclohexyl)-2-methoxyethanamine (PCMEA) and N-(1-phenylcyclohexyl)-2 ethoxyethanamine (PCEEA). After adding 250 micromol PCEEA x HCl (62 mg), a 1 l culture of CAD65 (S. pombe strain co-expressing human CYP reductase and CYP2B6) was fermented over 65 h (pH 8, 30 degrees C) and centrifuged. PCHEA and remaining parent drug were isolated from the supernatant by solid-phase extraction (SPE). The eluate was evaporated to dryness and reconstituted in HPLC solvent. Aliquots were separated by semi-preparative HPLC. From the respective fraction, PCHEA was extracted by liquid-liquid extraction and precipitated as hydrochloric salt. Approximately 80% of PCEEA was converted to PCHEA. The final yield of PCHEA x HCl was 9 mg (35 micromol). Its identity was confirmed by GC-MS, (1)H NMR and (13)C NMR. The product purity, as determined by HPLC-UV, was 95%. PMID- 19135817 TI - Stability of new potential ACE inhibitor in the aqueous solutions of different pH. AB - Angiotensin-converting enzyme (ACE) inhibitors are a group of active substances binding to an active site of ACE. Many authors who studied the structure activity relationship suggested the structural elements needed for a potent ACE inhibitor. While many authors studied the activity of ACE inhibitor substances only a few structure stability studies have been presented. In this paper the stability properties of molecule xPRIL were studied by determination of degradation path and rate of degradation in aqueous solutions with different pH (2.0, 6.8 and 12.0) and temperatures (40, 60 and 80 degrees C). The degradation of molecule through two main degradation paths was identified and confirmed by liquid chromatography and mass spectroscopy (LC-MS). Stability properties of xPRIL were determined in a stability study evaluated by high-performance liquid chromatography (HPLC). The first order kinetics of degradation reaction of xPRIL and Arrhenius equations for each pH were determined at observed conditions. xPRIL showed the highest stability at pH 2 solution. The degradation kinetics of xPRIL was compared to the degradation kinetics of enalapril maleate (EM) and perindopril (PER) in bio relevant solutions with pH 2.0 and 6.8. In addition to the stability study of xPRIL the forced degradation study of all three molecules at rigorous conditions was conducted. From the obtained results the structural element having the highest influence on stability properties of the studied molecules was identified. The fragmentation paths of xPRIL, its cyclization degradation product and its hydrolysis degradation product were identified and confirmed by MS/MS method. PMID- 19135818 TI - Determination of oxymatrine and its metabolite matrine in rat blood and dermal microdialysates by high throughput liquid chromatography/tandem mass spectrometry. AB - Oxymatrine (OMT) and matrine (MT) are the major quinolizidine alkaloids found in certain Sophora plants, which have been extensively used in China for the treatment of viral hepatitis, cancer, cardiac diseases and skin diseases (such as atopic dermatitis and eczema). A precise, sensitive and high throughput LC-MS/MS was developed to determine OMT and its metabolite MT in rat blood and dermis collected using microdialysis technique. Microdialysis probes were inserted into the jugular vein/right atrium and dermis of Wistar rats, and 3% OMT gel (1g) was administered via topical application. The samples were collected and then injected into the LC-MS/MS system after adding the internal standard (codeine, CDN). Chromatographic separation was achieved in a run time of 2min on a reversed phase short-column (50mmx2.1mm, 3.5microm). The mobile phase for column separation was methanol-ammonium formate (pH 5.0; 25mM) (70:30, v/v) with a flow rate of 0.3mL/min. A diverter valve was installed post-LC column for desalting. Detection of analytes and IS was done by tandem mass spectrometry, operating in positive ion and multiple reaction monitoring (MRM) acquisition mode. The protonated precursor to product ion transitions monitored for OMT, MT and IS was m/z 265.0-->247.3, 249.1-->148.3 and 300.0-->215.2, respectively. The lower limit of quantification (LLOQ) for OMT and MT was 0.5ng/mL. The calibration curves were linear over the range of 0.5-1000ng/mL for OMT and MT with a coefficient of determination >0.999. This selective and sensitive method is useful for the determination of OMT and MT and in the pharmacokinetic studies of these compounds. The blood and dermal concentration-time profile of OMT and its metabolite MT suggest that the limiting factor for dermal metabolism is the low capacity of enzymes in the skin rather than the quantity of penetrated OMT. PMID- 19135819 TI - Study of curcumin and genistein interactions with human serum albumin. AB - Curcumin, the yellow pigment from the rhizoma of Curcuma longa, is a widely studied polyphenolic compound which has a variety of biological activity as anti inflammatory and antioxidative agent. Genistein one of the flavonoids found in soybean and chickpeas inhibits DNA strand breaks acting as a direct scavenger of reactive oxygen species. Human serum albumin (HSA) with high affinity binding sites is a major transporter for delivering several endogenous compounds and drugs in vivo. The aim of this study was to examine the interactions of curcumin and genistein with human serum albumin at physiological conditions, using constant protein concentration and various pigment contents. FTIR, UV-Visible, CD and fluorescence spectroscopic methods were used to analyse drug binding mode, the binding constant and the effects of pigment complexation on HSA stability and conformation. Structural analysis showed that curcumin and genistein bind HSA via polypeptide polar groups with overall binding constants of K(curcumin)=5.5 (+/ 0.8)x10(4)M(-1) and K(genistein)=2.4 (+/-0.40)x10(4)M(-1). The number of bound pigment (n) is 1.33 for curcumin and 1.49 for genistein. The HSA conformation was altered by pigment complexation with reduction of alpha-helix and increase of random coil and turn structures suggesting a partial protein unfolding. PMID- 19135820 TI - Extraction and determination of some psychotropic drugs in urine samples using dispersive liquid-liquid microextraction followed by high-performance liquid chromatography. AB - A simple, rapid and sensitive method termed as dispersive liquid-liquid microextraction (DLLME) combined with high-performance liquid chromatography ultraviolet detector (HPLC-UV) has been proposed for the determination of three psychotropic drugs (amitryptiline, clomipramine and thioridazine) in urine samples. The determination was performed on a C(8) column under the optimal chromatographic conditions (mobile phase: ammonium acetate (0.03 mol L(-1), pH 5.5)-acetonitrile (60:40, v/v); flow rate: 1.0 mL min(-1); detection wavelength: 238 nm). Several factors influencing the extraction efficiency of the target drugs, such as pH, extraction and disperser solvent type and their volume, extraction time and ion strength were studied and optimized. Under the optimal DLLME conditions, the absolute recoveries of amitryptiline, clomipramine and thioridazine from the urine samples were 96, 97 and 101%, respectively. The detection limits (LODs) and quantification (LOQs) of the proposed approach were 3 and 10 ng mL(-1) for amitryptiline, 7 and 21 ng mL(-1) for clomipramine, and 8 and 25 ng mL(-1) for thioridazine, respectively. The relative standard deviations (RSDs) for nine replicate determinations at 0.100 microg mL(-1) level of target drugs were less than 4.8%. Good linear behaviors over the investigated concentration ranges were obtained with the values of R(2)>0.998 for the target drugs. The proposed method was successfully applied to the real urine samples from two female patients under amitryptiline and clomipramine treatment, respectively. PMID- 19135821 TI - Antibacterial activity of telithromycin and comparators against pathogens isolated from patients with community-acquired respiratory tract infections: the Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin study year 5 (2003-2004). AB - The activity of telithromycin and comparator antibacterials was examined in isolates of Streptococcus pneumoniae and Haemophilus influenzae isolated from patients with community-acquired pneumonia (CAP), acute exacerbations of chronic bronchitis (AECB), or sinusitis during year 5 (2003-2004) of the Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin global resistance surveillance study. Among S. pneumoniae, penicillin nonsusceptibility and erythromycin resistance were 35.7% and 36.0%, respectively. beta-Lactamase was produced by 12.3% of H. influenzae isolates. beta-Lactamase negative ampicillin-resistant strains, mainly from Japan, comprised 5.2% of global H. influenzae isolates. Telithromycin and levofloxacin were the most active agents tested against S. pneumoniae and H. influenzae (>99% of isolates susceptible) isolated from patients with CAP, AECB, or bacterial sinusitis. Amoxicillin-clavulanate, levofloxacin, and telithromycin were the most active agents against multidrug-resistant S. pneumoniae. PMID- 19135822 TI - What do Nepalese medical students and doctors think about patient-centred communication? AB - OBJECTIVE: To ascertain the attitudes of Nepalese medical students and doctors regarding aspects of doctor-patient communication. METHOD: A cross-sectional survey, using the Patient-Practitioner Orientation Scale (PPOS), was undertaken with students and doctors in a teaching hospital in rural Nepal. Qualitative research was also done, using semi-structured interviews and focus groups, with a sample from the same population. The author's participant observations provided a third data source. RESULTS: Participants generally expressed 'patient-centred' attitudes, particularly in the following areas: power-sharing; the importance of social context; friendliness; providing adequate time and explanations. They thought that the hierarchical nature of society carried over into medical practice, particularly noting the exalted position of doctors and the importance of social and financial issues in patient outcomes. The participants thought that Nepalese doctors currently practice in a 'doctor-centred' manner but thought that this should change. CONCLUSION: There was a marked contrast between the attitudes expressed by participants and the way that they see medicine is currently practiced. The results also showed an under-appreciation of Nepalese patients' universal desire for being given good information. PRACTICE IMPLICATIONS: This study underlines the need for an increased understanding of local preferences regarding medical communication styles. Methods of providing information to patients need to be strengthened in clinical practice. PMID- 19135823 TI - Shifting goals in medical communication. Determinants of goal detection and response formation. AB - OBJECTIVES: Research and education of medical communication different perspective, making the extraction of clear recommendations from research that can be applied in education not always possible. In education, medical communication is encountered from a goal-oriented perspective, which is often lacking in quantitative research where the relationship between process variables and the content of medical practice is often ignored. The aim of this paper is to bring the worlds of research and education together by presenting a comprehensive model of determinants explaining the behavior of physicians in daily practice. A basic notion in this model is that medical communication is goal-oriented, problem-solving behavior. Goals in communication are not fixed, but permanently changing over time. Hence, communication abilities do not rely on behavioral skills only but also on perceptual skills in identifying goals. A number of determinants affect the cognitive processes of goal appraisal and response formation: knowledge, attitudes, social norms, self-efficacy, stressors and interfering goals. CONCLUSION: Modeling medical communication as goal-oriented problem-solving behavior, and recognizing the complexity of goal appraisal and other key determinants of response formation may provide a common focus for both research and education in measuring, explaining and improving the HCP's behavior. PRACTICE IMPLICATIONS: In education not only skill practicing but also reflection on the process and outcomes is important to understand how one acts in practice situations and should act in future situations. In research measurements should be expanded to take contextual and goal-oriented dimensions of the process of communication into account to make findings more relevant for education and practice. PMID- 19135824 TI - Patients' reflections on communication in the second-opinion hematology-oncology consultation. AB - OBJECTIVE: The nature of communication between patients and their second-opinion hematology consultants may be very different in these one-time consultations than for those that are within long-term relationships. This study explored patients' perceptions of their second-opinion hematology-oncology consultation to investigate physician-patient communication in malignant disease at a critical juncture in cancer patients' care and decision-making. METHODS: In-depth telephone interviews with a subset of 20 patients from a larger study, following their subspecialty hematology consultations. RESULTS: Most patients wanted to contribute to the consultation agenda, but were unable to do so. Patients sought expert and honest advice delivered with empathy, though most did not expect the consultant to directly address their emotions. They wanted the physician to apply his/her knowledge to the specifics of their individual cases, and were disappointed and distrustful when physicians cited only general prognostic statistics. In contrast, physicians' consideration of the unique elements of patients' cases, and demonstrations of empathy and respect made patients' feel positively about the encounter, regardless of the prognosis. CONCLUSIONS: Patients provided concrete recommendations for physician and patient behaviors to enhance the consultation. PRACTICE IMPLICATIONS: Consideration of these recommendations may result in more effective communication and increased patient satisfaction with medical visits. PMID- 19135825 TI - Communication skills training for general practitioners to promote patient coping: the GRIP approach. AB - OBJECTIVE: To develop, perform and test the effects of a communication skills training program for general practitioners (GPs). The program specifically addresses the patients' coping and resources despite more or less severe psychological or physical illness. METHODS: A training model was developed, based on cognitive therapy and solution-focused therapy. The training was given the acronym GRIP after its main content: Get a measure of the patient's subjective complaints and illness attributions. Respond to the patient's understanding of the complaints. Identify resources and solutions. Promote positive coping. The study involved a quasi-experimental design in which 266 consultations with 25 GPs were video recorded. Forty hours of communication skills training were given to the intervention group. RESULTS: Consultation duration, patient age and distress determined the frequency of the GRIP communication. There was a significant effect of training on four particular subcategories of the GRIP techniques. The effect of the training was most evident in a subgroup of GPs who used little or no resource-oriented communication before training. CONCLUSION: This pilot training model may help change the GPs' communicative pattern with patients in some situations. PRACTICE IMPLICATION: Communication skills training programmes that emphasize patient attributions and personal resources should be developed further and tested in general practice settings with an aim to promote patient coping. PMID- 19135826 TI - Do physicians improve their communication skills between finishing medical school and completing internship? A nationwide prospective observational cohort study. AB - OBJECTIVE: To test whether young physicians improve their communication skills between graduating from medical school and completing clinical internship, and to explore contributing background and/or internship factors. METHODS: Norwegian medical students graduating June 2004 were invited to take part in a videotaped standardized patient interview February 2004. Of the 111 students who originally participated, 62 completed a second interview February 2006. Observed communication skills were assessed with the Arizona Communication Interview Rating Scale (ACIR). RESULTS: The level of communication skills increased significantly during the period for participants overall; and for females but not males. General social skills reached significantly higher levels than specific professional skills, both types of skill improving during the study. Independent predictors were working in local hospitals, learning atmosphere and low stress. At school completion, 50% reached a level defined as 'advanced beginner'. Towards the end of the internship, 58% reached 'capable' and 27% 'competent' levels of communication skills. CONCLUSIONS: Female physicians improved most in communication skills, the gender difference being multivariate mediated through low stress levels and learning atmosphere. The findings support the division of communication skills into general social and specific professional skills. PRACTICE IMPLICATIONS: The relatively low proportion of young physicians, especially males, developing the capability to practise independently at internship completion indicates a need for more effective training in communication skills, during both medical school and internship. PMID- 19135827 TI - Theories in health communication research. PMID- 19135828 TI - Rapid calcitonin response to experimental hypercalcemia in healthy horses. AB - Calcium has important physiological functions, and disorders of calcium homeostasis are frequent in horses. We have made important progress understanding equine calcium homeostasis; however, limited information on equine calcitonin (CT) is available, in part because of the lack of validated CT assays. To determine the CT response to high ionized calcium (Ca(2+)) concentrations in healthy horses, we induced hypercalcemia in 10 healthy horses using a calcium gluconate 23% solution (5mg/kg; 120 mL/500 kg horse) infused over 4 min. Four horses were infused with 120 mL of 0.9% NaCl and used as controls. We validated a human-specific CT radioimmunoassay for use in horses. Serum Ca(2+) concentrations increased from 6.2+/-0.3mg/dL to 9.9+/-0.5mg/dL (4 min; P<0.01). Serum CT increased from 16.7+/-8.0 pg/mL to 87.1+/-55.8 pg/mL at 2 min, and 102.5+/-51.1 pg/mL at 4 min (P<0.01). Serum CT returned to baseline by 20 min, whereas serum Ca(2+) returned to baseline by 40 min. Of interest, CT concentrations returned to baseline despite hypercalcemia, suggesting thyroid gland C-cell CT depletion. Resting CT values higher than 40 pg/mL were considered abnormally elevated. No significant changes in serum Ca(2+) or CT concentrations were found in control horses. The coefficients of variation for the CT radioimmunoassay were lower than 11.9%. We conclude that the equine thyroid gland C-cell responds quickly to changes in extracellular Ca(2+) concentrations by secreting large quantities of CT into the systemic circulation, indicating that CT is important in equine calcium homeostasis. The human CT radioimmunoassay can be used to measure changes in equine CT. PMID- 19135829 TI - Late conversion of aortic stent grafts. AB - OBJECTIVES: The frequency of late removal of endovascular abdominal aortic repair (EVAR) parallels the rise of endovascular aortic repair. Evaluation of outcomes for EVAR explants may identify risks for complications and alter clinical management. METHODS: A patient database was used to identify EVAR patients requiring explant >1 month after implant. A retrospective analysis was conducted of the type of graft, duration of implant, reason for removal, operative technique, death, and length of stay. RESULTS: During 1999 through 2007, 1606 EVARs were performed, and 25 patients required explantation, with an additional 16 referred from other institutions (N = 41). The average age was 73 years (range, 50-87 years); 90% were men. Grafts were excised after a median of 33.3 months (range, 3-93 months). Explanted grafts included 16 AneuRx (40%), 7 Ancure (17%), 6 Excluder (15%), 4 Zenith (10%), 4 Talent (10%), 1 Cook Aortomonoiliac rupture graft, 1 Endologix, 1 Quantum LP, and 1 homemade tube graft. Overall hospital mortality was 19% and occurred after conversion for rupture in 4, and in infected graft, aortoenteric fistula, repair of new aneurysm of the visceral segment, and claudication due to graft stenosis in one patient each. Elective EVAR-related mortality was 3.3%. Mortality was higher in patients with rupture compared with nonrupture (4 of 6 vs 3 of 35, P 30). Period 2 patients when compared with period 1 had more technically challenging anatomy with more calcified lesions (68% vs 27%), longer lesions (15.9 mm vs 8.2 mm), and higher incidence of ulceration (55% vs 27%) (all P < .04). CONCLUSION: Despite successful performance of 47 consecutive CAS procedures without permanent neurologic sequelae, significant reductions in periprocedural embolic events as identified via DW-MRI lesions may be achieved through implementation of quality improvement measures identified through continuous outcome analysis. The long term neurologic benefits associated with reduced subclinical neurologic events remains to be determined. PMID- 19135834 TI - Recruiting participants with peripheral arterial disease for clinical trials: experience from the Study to Improve Leg Circulation (SILC). AB - OBJECTIVE: To describe the success of diverse recruitment methods in a randomized controlled clinical trial of exercise in persons with peripheral arterial disease (PAD). METHODS: An analysis of recruitment sources conducted for the 746 men and women completing a baseline visit for the study to improve leg circulation (SILC), a randomized controlled trial of exercise for patients with PAD. For each recruitment source, we determined the number of randomized participants, the rate of randomization among those completing a baseline visit, and cost per randomized participant. RESULTS: Of the 746 individuals who completed a baseline visit, 156 were eligible and randomized. The most frequent sources of randomized participants were newspaper advertising (n = 67), mailed recruitment letters to patients with PAD identified at the study medical center (n = 25), and radio advertising (n = 18). Costs per randomized participant were $2750 for television advertising, $2167 for Life Line Screening, $2369 for newspaper advertising, $3931 for mailed postcards to older community dwelling men and women, and $5691 for radio advertising. Among those completing a baseline visit, randomization rates ranged from 10% for those identified from radio advertising to 32% for those identified from the Chicago Veterans Administration and 33% for those identified from posted flyers. CONCLUSION: Most participants in a randomized controlled trial of exercise were recruited from newspaper advertising and mailed recruitment letters to patients with known PAD. The highest randomization rates after a baseline visit occurred among participants identified from posted flyers and mailed recruitment letters to PAD patients. PMID- 19135836 TI - Fenestrated endovascular repair for juxtarenal aortic pathology. AB - OBJECTIVE: To evaluate the outcomes after fenestrated endovascular aortic repair (f-EVAR) in a tertiary European referral center. METHODS: All patients treated with commercially available custom-made f-EVAR between September 2002 and June 2007 were prospectively enrolled in a computerized database including co morbidities and aneurysm morphology. Patients were retrospectively analyzed. Follow-up consisted of clinical examinations and computed tomography (CT) scanning. RESULTS: A total of 54 patients were included in this study. Median age was 72 (interquartile range [IQR] 68-76) years and 85% were men. Median preoperative aneurysm diameter was 60 (53-66) mm. One hundred thirty-four vessels were targeted (43 scallops, 91 fenestrations) and 96 stents were placed (69 bare, 27 covered). Target vessel catheterization was achieved in 98% of cases. Two patients (3.7%) died within 30 days, 1 from trash embolization and multiorgan failure and 1 from retroperitoneal bleeding caused by a renal artery perforation. Three type I endoleaks occurred intraoperatively, two sealed pre-discharge and one was treated with a Palmaz stent (Cordis, Miami Lakes, Fla) on postoperative day 4. Thirteen patients had type II endoleaks, and 2 required treatment. The median clinical follow-up was 25 (12-32) months with median CT follow-up of 22 (4 26) months. Aneurysm diameter decreased >or=5 mm in 47%, was unchanged in 50%, and increased >or=5 mm in 3% of patients at 1 year. There were three type II endoleaks at 1-year follow-up, one of which was successfully treated after 19 months due to aneurysm growth. Ninety-six percent of target vessels remained patent during the study period and all occlusions occurred within the first year of follow-up. Five target vessels occluded (2 renal arteries [RAs] and 3 superior mesenteric arteries [SMAs]) without symptoms during follow-up and successful reinterventions were done on 2 stenosed RAs. Three patients suffered creatinine increase but none needed dialysis. One late aneurysm-related death occurred due to massive bleeding during redo surgery for infection. CONCLUSION: Despite complex anatomy or severe comorbidities in these patients f-EVAR has acceptable short- and midterm results in this series which includes a learning curve and offers a valid treatment alternative to patients unsuitable for standard EVAR or open repair. PMID- 19135837 TI - Prospective randomized trial of operative vs interventional treatment for renal artery ostial occlusive disease (RAOOD). AB - INTRODUCTION AND OBJECTIVES: Patients with either renovascular hypertension (RVH) and/or renal insufficiency (RI) due to renal artery ostial occlusive disease (RAOOD) can successfully undergo an open surgical reconstruction procedure (OSRP), but since the publication of Blum et al(1) percutaneous balloon stent angioplasty (PTRA + stent) leaving a small part of the stent within the aorta has become very popular. However, balloon dilatation and stenting does not remove the atherosclerotic plaque, which is often heavily calcified but leads to disruption of the plaque causing myointimal hyperplasia and recurrent stenosis. Therefore, a comparison of the two treatment modalities concerning complications and durability in a prospective randomized design was felt to bring more insight to the discussion. METHODS: From 1998 to 2004, we performed OSRP in 330 patients with RVH and/or RI for various locations of RAOOD. During this time period, 50 patients (female 18, male 32, mean age 64.4 years) with RAOOD of at least 70% stenosis (DSA and duplex criteria) in one or both renal arteries, who did not require other aorto/mesenteric/iliac reconstructive procedures agreed and were randomized to either OSRP (n = 25 patients, 49 arteries) or PTRA + stent (n = 25 patients, 28 arteries). Two patients crossed over to surgical treatment. Patients were followed on a regular basis for 4 years and longer. Endpoints were re occurrence of RAOOD and impairment of either kidney function or RVH. RESULTS: We approached 77 arteries. There was no early mortality in either group, but directly procedure-related morbidity was 13% in the interventional group and 4% in the surgical group. Four-year follow-up mortality was 18% in the interventional group and 25% in the surgical group. Both groups showed significant improvement of RVH (P < .001 in each group) as well as improvement or stabilization in patients with insufficient renal function. Freedom from recurrent RAOOD (>70%) was achieved in 90.1% of the surgical group and 79.9% of the interventional group. CONCLUSION: Both treatment modalities showed good early results concerning RVH, kidney function, and renal perfusion. Despite a higher number of bilateral renal artery reconstructions in patients undergoing OSRP, which was probably due to the preferred technique of transaortic endarterectomy eliminating the plaque originating in the aorta and usually extending into both renal arteries, mortality was not higher and procedure-related morbidity was even lower compared to PTRA + stent. These findings and also longer durability of OSRP imply that surgical reconstruction remains the gold standard for patients with RAOOD before PTRA + stent may be considered. PMID- 19135838 TI - Endoscopically directed external support of femoral vein valves. AB - BACKGROUND: This study investigated the optimization of indications to external valve support using the Vedensky Spiral for primary deep venous reflux based on the femoral vein proximal (ostial) valve anatomy type. METHODS: The external Vedensky Spiral was used for correction of valvular insufficiency in 28 extremities of 24 patients (18 women, 6 men) during 1998 to 2002. Patients were a mean age of 54.6 years (range, 32-76 years). The clinical manifestation was C(4) in 10 limbs and C(5) in 18 limbs. Primary axial deep reflux was present in all 28 extremities, and axial superficial reflux was also present in 26. Duplex scanning and descending phlebography were used to estimate the functional condition of the deep vein valves. Fibrophleboscopy intraoperatively to study the anatomic status of the femoral vein valves and for checking the repaired valve function was used. RESULTS: Phleboscopy showed the following anatomy of ostial valves of the femoral vein: 16 valves had wide separation of cusps, 11 had elongation of cusps, and one had a monocusp. The competence of femoral vein ostial valves was completely restituted in all cases with wide separation of cusps. Multiple corrections were performed in cases with incomplete competence of the ostial valve (valve with elongation of cusps or monocusp). CONCLUSION: External valve support by the Vedensky Spiral is an effective and simple method of correction of incompetent femoral vein valves in limbs with primary deep venous reflux. The technical success of the intervention depends on the type of valve insufficiency and the correct choice of spiral diameter. PMID- 19135839 TI - Contrast-enhanced ultrasound versus color duplex ultrasound imaging in the follow up of patients after endovascular abdominal aortic aneurysm repair. AB - PURPOSE: This study assessed the negative predictive value, sensitivity, specificity, and diagnostic accuracy of real-time contrast-enhanced ultrasound imaging (CEUS) in the detection of endoleaks in patients with abdominal aortic aneurysm (AAA) who underwent endovascular repair (EVAR) compared with unenhanced ultrasound imaging. Computed tomography angiography (CTA) was the gold standard. The secondary objective was to define the optimal dose of the second-generation contrast agent to routinely use in the CEUS examinations for endoleak detection. METHODS: The study enrolled 84 patients with unruptured AAA who were treated with EVAR and underwent CTA follow-up. In the same day, CTA (4- x 1-mm collimation, 1.25-mm slice width), unenhanced US imaging and CEUS imaging was performed in all patients. The CEUS studies were performed after an intravenous bolus injection of 1.2 mL and 2.4 mL of a second-generation contrast agent with continuous low mechanical index (range, 0.01-0.04) real-time tissue harmonic imaging. The unenhanced US and CEUS studies were interpreted separately by two independent experienced readers to detect the presence of endoleaks by viewing recorded videotapes according to a five-point confidence scale. The standard of reference was represented by the consensus reading of CTA performed by two experienced radiologists not involved in the image analysis. Qualitative analysis as well as sensitivity, specificity, negative predictive value, and diagnostic accuracy in detecting endoleaks of each reading session were compared. RESULTS: CEUS imaging significantly improved the diagnostic performance of unenhanced US studies in the detection of endoleaks in terms of sensitivity (97.5% vs 62.5%), negative predictive value (97.3% vs 65.1%), accuracy (89.3% vs 63.1%), and specificity (81.8% vs 63.6%). The optimal dose of contrast agent to detect and characterize endoleaks was 2.4 mL. No adverse events were recorded during the study. CONCLUSIONS: The results showed CEUS imaging is a fast, noninvasive, reliable, and valid alternative to multislice CTA for endoleak detection in endovascular aortic stent graft patients, and is superior to unenhanced US imaging. Contrast enhanced ultrasound imaging should be performed using a recommended contrast medium dose of 2.4 mL. PMID- 19135841 TI - Treatment of ruptured abdominal aortic aneurysm after endovascular abdominal aortic repair: A comparison with patients without prior treatment. AB - OBJECTIVE: A retrospective analysis of immediate outcomes following aneurysm rupture (rAAA) in two groups: patients previously treated at our center with primary endovascular repair (EVAR) and patients without previous EVAR for abdominal aortic aneurysms (AAA) in an 8-year period. METHODS: Fourteen patients with a confirmed rAAA identified throughout the follow-up period following primary EVAR repair at our center (from a population of 820 AAA treated at our center in election) were retrospectively compared with 155 patients without previous EVAR in the same time period, from the introduction of an intention-to treat protocol with EVAR for rAAA in January 1999. Primary study outcomes included 30-day mortality and severe systemic complications following rAAA correction with both open and EVAR treatments. RESULTS: In the 14 patients secondary interventions were necessary throughout follow-up prior to rupture in 43% (6/14). The mean time to rupture was 50.23 months (9-113). The mean increase in maximum aneurysmal diameter at rupture was 18.39 mm. Type of endoleaks observed at rupture: 35.7% I proximal, 35.7% III contralateral stump disconnection, 14.3% I distal, 14.3% III midgraft tear: treatment at rupture included five EVAR corrections with aortouniiliac endografts, four EVAR corrections with extensions, and five surgical conversions. Thirty-day mortality between the two groups, 28.5% (patients with prior EVAR) 38.7% (patients without prior EVAR), and severe systemic complications, 50% vs 37.6%, were not found to be statistically significant. Hemodynamic instability, 36% (patients with prior EVAR) 63% (patients without prior EVAR), was found to be an independent predictor of 30-day mortality (P < .0001), whereas severe systemic complications, 50% vs 33.5%, did not influence the same outcome (P = .852). CONCLUSIONS: In terms of mortality, it would be logical to expect a protection from the endograft in patients with previous EVAR. A trend seems to confirm this assumption, but no statistical significance was found, which may be due to the small population size. PMID- 19135843 TI - Decrease in total aneurysm-related deaths in the era of endovascular aneurysm repair. AB - OBJECTIVE: With the expansion of elective abdominal aortic aneurysm (AAA) repair after the introduction of endovascular aneurysm repair (EVAR), there is a concern that even with a lower operative mortality there could be an increasing number of aneurysm-related deaths. To evaluate this, we looked at national trends in AAA repair volume as well as mortality rates after intact and ruptured AAA repair encompassing the introduction of EVAR. METHODS: Patients with intact or ruptured AAA undergoing open repair or EVAR and all those with a diagnosis of ruptured AAA were identified within the 1993 to 2005 Nationwide Inpatient Sample database using International Classification of Diseases, 9th Revision, diagnosis and procedure codes. The number of repairs, number of rupture diagnoses without repair, number of deaths, and associated mortality rates were measured for each year of the database. Outcomes (mean annual volumes) were compared from the pre EVAR era (1993 to 1998) with the post-EVAR era (2001 to 2005). RESULTS: Since its introduction, EVAR increased steadily and accounted for 56% of repairs yet only 27% of the deaths for intact repairs in 2005. The mean annual number of intact repairs increased from 36,122 in the pre-EVAR era to 38,901 in the post-EVAR era, whereas the mean annual number of deaths related to intact AAA repair decreased from 1693 pre-EVAR to 1207 post-EVAR (P < .0001). Mortality for all intact AAA repair decreased from 4.0% to 3.1% (P < .0001) pre-EVAR and post-EVAR, but open repair mortality was unchanged (open repair, 4.7% to 4.5%, P = .31; EVAR, 1.3%). During the same time, the mean annual number of ruptured repairs decreased from 2804 to 1846, and deaths from ruptured AAA repairs decreased from 2804 to 1846 (P < .0001). Mortality for ruptured AAA repair decreased from 44.3% to 39.9% (P < .0001) pre-EVAR and post-EVAR (open repair, 44.3% to 39.9%, P < .001; EVAR, 32.4%). The overall mean annual number of ruptured AAA diagnoses (9979 to 7773, P < .0001) and overall mean annual deaths from a ruptured AAA decreased post-EVAR (5338 to 3901, P < .0001). CONCLUSION: Since the introduction of EVAR, the annual number of deaths from intact and ruptured AAA has significantly decreased. This coincided with an increase in intact AAA repair after the introduction of EVAR and a decrease in ruptured AAA diagnosis and repair volume. PMID- 19135845 TI - [Pericarditis in systemic lupus erythematosus: prevalence and clinical and immunologic characteristics]. AB - PURPOSE: Our study sought to determine the frequency and clinical and immunologic characteristics of pericarditis in patients with systemic lupus erythematosus (SLE). METHODS: We retrospectively analyzed the files of all patients with SLE diagnosed between 1987 and 2005. The study included only those patients who had undergone echocardiography. They were divided in two groups according to whether or not they had pericarditis, and their clinical and immunologic characteristics and outcome were compared. RESULTS: Our records found 180 patients with SLE, 97 of whom had had echocardiography. The frequency of pericarditis was 39%. The average age of the 34 women and 4 men with pericarditis at its onset was 36.4 years. There were 59 SLE patients without pericarditis, 56 women and 3 men with an average age of 28.8 years. Pericarditis was frequently associated with pleural effusions. The prevalence of anticardiolipin antibodies was significantly lower in the group of patients with pericarditis. CONCLUSION: Pericarditis is the most common cardiac manifestation in SLE. Its frequency in this sample of Tunisian patients was similar to that described in different ethnic groups. The association between pericarditis and pleuritis is standard. Anticardiolipin antibodies, commonly positive in valve diseases, were less frequent in our patients with pericarditis. PMID- 19135846 TI - [Biological diagnosis of pheochromocytomas and paragangliomas]. AB - Pheochromocytomas and/or paragangliomas are rare, heterogeneous tumors of the chromaffin cells. Thirty percent of the patients presented with these diseases in a hereditary context. The biological diagnosis relies on the identification of excessive secretion of the metanephrines which are more sensitive and specific than those of catecholamines The published recommendations give the opportunity to choose between the free metanephrines and the fractionated metanephrines in sera or urines. The concentrations of the free plasmatic metanephrines reflect the ongoing production of tumor. They are little sensitive to the renal failure. The assay of the vanillylmandelic acid should be dropped because of its inefficiency. The assay of the chromogranin A in serum should be used in association with those of metanephrines in the diagnosis but also in the follow up. Its role still has to be precised. PMID- 19135847 TI - [Pathogenesis of chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is a respiratory disease characterized by permanent and progressive airway obstruction. Cigarette smoking is the main cause responsible for COPD although only 15 to 25 % of smokers develop COPD. Mechanisms underlying COPD pathogenesis are not fully understood. Structural abnormalities in small airways (bronchioles < 2mm in diameter) are the main determinants of airway obstruction; obstruction of these bronchioles is related to increase in airway wall thickness (associated with peribronchiolar fibrosis) and to plugging by mucus exudates. Alveolar wall destruction (emphysema) also contributes to airway obstruction and to gas exchange abnormalities. Current knowledge related to molecular and cellular mechanisms responsible for these structural modifications are reviewed. PMID- 19135848 TI - [Antimalarial's retinopaty remains a current threat]. PMID- 19135849 TI - [Effect of physical activity on anxiety and depression]. AB - The advantages of physical activity are widely recognised from both a physiological and a psychological perspective. Evidence seems to demonstrate that physical activity is associated with decreases in depression and anxiety in clinical and non-clinical populations. There are a number of physiological, biochemical and psychological explanations which should be considered to understand the psychological effects of exercise. Physical activity may be considered as an adjunct to psychotherapeutic and pharmacological treatments of depression and anxiety. Physical activity appears to be a non-specific form of treatment with psychotherapeutic potential that should not be ignored. PMID- 19135851 TI - The effect of an artificially lengthened vocal tract on estimated glottal contact quotient in untrained male voices. AB - The use of hard-walled narrow tubes, often called resonance tubes, for the purpose of voice therapy and voice training has a historical precedent and some theoretical support, but the mechanism of any potential benefit from the application of this technique is not well understood. Fifteen vocally untrained male participants produced a series of spoken /a/ vowels at a modal pitch and constant loudness, before and after a minute of repeated phonation into a 50-cm hard-walled glass tube at the same pitch and loudness targets. Electroglottography was used to measure the glottal contact quotient (CQ) during each phase of the experiment. Single-subject analysis revealed statistically significant changes in CQ during tube phonation, but with no discernable pattern across the 15 participants. These results indicate that the use of resonance tubes can have a distinct effect on glottal closure, but the mechanism behind this change remains unclear. The implication is that vocal loading techniques such as this need to be studied further with specific attention paid to the underlying mechanism of any measured changes in glottal behavior, and especially to the role of instruction and feedback in the therapeutic and pedagogical application of these techniques. PMID- 19135850 TI - Endothelin receptor antagonism by tezosentan attenuates lung injury induced by aortic ischemia-reperfusion. AB - Tezosentan is a novel dual endothelin receptor antagonist. The purpose of this study was to examine the effect of tezosentan on lung injury induced by abdominal aortic ischemia-reperfusion (IR) in rats. Thirty-two Wistar-albino rats were randomized into four groups (eight per group) as follows: control group (sham laparotomy), aortic IR group (120 min ischemia and 120 min reperfusion), aortic IR + tezosentan group (a bolus intravenous injection of 10 mg/kg tezosentan before ischemia plus continuous intravenous infusion of 1 mg/kg/hr tezosentan during 120 min ischemia and 120 min reperfusion), and control + tezosentan. Blood and lung tissue samples were obtained for biochemical analysis. Protein concentrations in bronchoalveolar lavage fluid and lung wet/dry weight ratios were measured. A histological evaluation was also done. Aortic IR significantly increased (p < 0.05 vs. control group) and tezosentan significantly decreased (p < 0.05 vs. aortic IR group) the plasma level of tumor necrosis factor-alpha; lung tissue levels of malondialdehyde, catalase, and myleperoxidase; and protein concentration in bronchoalveolar lavage fluid and lung wet/dry weight ratio. Histological evaluation showed that tezosentan attenuated the morphological changes associated with lung injury. The results of this study indicate that tezosentan attenuates lung injury induced by aortic IR in rats. We propose that this protective effect of tezosentan is due to (1) reduced systemic inflammatory response, (2) reduced oxidative stress and lipid peroxidation in lung tissue, (3) reduced pulmonary microvascular leakage, and (4) inhibition of leukocyte infiltration into lung tissue. PMID- 19135852 TI - Influence of abusive vocal habits, hydration, mastication, and sleep in the occurrence of vocal symptoms in teachers. AB - Some vocal disorders in teachers are associated with occupational factors, but there are few studies that analyze the influence of vocal habits, fluid intake, mastication, and sleep on these disorders. The objective was to analyze the occurrence of vocal fatigue, hoarseness, and dry throat in elementary and high school teachers and their association with vocal habits, fluid intake, mastication, and sleep. A sample of 422 elementary and secondary school teachers was studied using a specific questionnaire. The multiple regression analysis showed that hoarseness was associated with absence of water intake (odds ratio (OR)=1.7; P=0.047), yelling/speaking loudly (OR=1.6; P=0.058), jaw-opening limitations (OR=3.8; P=0.003), average of 6 hours of sleep/night (OR=1.7; P=0.039), and waking-up feeling replenished (OR=2.0; P=0.020). The presence of vocal fatigue was significantly associated with yelling/speaking loudly (OR=2.2; P=0.013), speaking excessively (OR=2.4; P=0.023), difficulty to open the mouth to masticate (OR=6.6; P=0.003), less than 6 hours of sleep (OR=4.0; P=0.008), and waking-up feeling replenished (sometimes OR=2.8; P=0.003; or never OR=3.3; P=0.002). The presence of dry throat was associated with being a former smoker (OR=3.3; P=0.011) and having jaw-opening limitations (OR=3.9; P=0.021). In recent years, speech and hearing interventions with teachers have focused on health-care promotion actions and prevention of vocal disorders, prioritizing issues related with hydration and healthy vocal use habits. However, the findings in the present study show the need to further focus on lifestyle habits related to sleep and eating habits. PMID- 19135853 TI - Development and validation of a voice disorder outcome profile for an Indian population. AB - No instrument exists for the people of India to measure outcomes of voice disorders. The objective of this study was to develop a statistically robust tool for assessing voice disorder outcomes in the Indian population. A 32-item assessment tool called the Voice Disorder Outcome Profile (Voice-DOP) was developed after consultation with two different sets of speech-language pathologists and individuals with voice disorders. Voice-DOP measures voice disorder outcomes in three domains, physical, emotional, and functional. The questionnaire was given to 42 individuals with various voice disorders and 30 control subjects with no vocal pathology. The data obtained were subjected to measures of reliability (internal consistency and test-retest) and validity (content, construct, and concurrent). Results showed that Voice-DOP had high internal consistency (Cronbach's alpha levels from 0.49 to 0.84) and test-rest reliability (r=0.96-0.99). Voice-DOP differentiated the dysphonic group from the control group, and correlations of r=0.49-0.87 were obtained between the total Voice-DOP scores and the domain scores, findings suggesting adequate construct validity. Concurrent validity was supported by a significant correlation (r=0.51) between the Voice-DOP scores and the self-perception of severity by the individuals with dysphonia. A comparison of Voice-DOP scores between males and females revealed no significant difference in their outcomes. The conclusion of this study is that Voice-DOP is a sufficiently reliable and valid tool to measure voice disorder outcomes in the Indian population. PMID- 19135854 TI - The effectiveness of the glottal to noise excitation ratio for the screening of voice disorders. AB - This paper evaluates the capabilities of the Glottal to Noise Excitation Ratio for the screening of voice disorders. A lot of effort has been made using this parameter to evaluate voice quality, but there do not exist any studies that evaluate the discrimination capabilities of this acoustic parameter to classify between normal and pathological voices, and neither are there any previous studies that reflect the normative values that could be used for screening purposes. A set of 226 speakers (53 normal and 173 pathological) taken from a voice disorders database were used to evaluate the usefulness of this parameter for discriminating normal and pathological voices. To evaluate this parameter, the effect of the bandwidth of the Hilbert envelopes and the frequency shift have been analyzed, concluding that a good discrimination is obtained with a bandwidth of 1000 Hz and a frequency shift of 300 Hz. The results confirm that the Glottal to Noise Excitation Ratio provides reliable measurements in terms of discrimination among normal and pathological voices, comparable to other classical long-term noise measurements found in the literature, such as Normalized Noise Energy or Harmonics to Noise Ratio, so this parameter can be considered a good choice for screening purposes. PMID- 19135855 TI - Hoarseness as a sign of possible nonspecific mucosal hyperreactivity in vocal tract. AB - The aim of the present paper was to evaluate the possible impact of methacholine challenge on the voice and vocal tract in 10 patients reporting hoarseness due to presumable hyperreactivity to some environmental factors. Ten age and gender matched healthy subjects served as controls. Methacholine was used for hypersensitivity challenge and saline solution (0.9% NaCl) as control substance. Subjects were examined on two separate challenge occasions testing either methacholine in increasing doses (3, 6, 12 mg) or NaCl. Voice recordings, videolaryngoscopy, and measurement of nasal secretion were performed before and after each sniffing session. Subjective complaints were documented. Data were statistically analyzed with three-way analysis of variance and group comparisons performed. Perceptual analysis of voice recordings showed an increase of the grade of voice disorder in patients after exposure to both substances. Moreover, one of the patients became aphonic and another severely dysphonic after NaCl. No voice quality changes were detected in controls. The videolaryngoscopy findings divergent from normal did not increase in any group. Nasal secretion was significantly higher after sniffing of methacholine than NaCl in both groups. The frequency of subjective complaints was equal in both groups after both substances. However, there was a qualitative difference in the character of the symptoms: the patients complained of throat, vocal, and nasal symptoms whereas the controls complained exclusively of nasal symptoms. The study supports the view that vocal dysfunction after exposure to non-specific environmental irritating factors may be triggered mainly by emotional mechanisms such as off warding reaction or dissociative disorder. PMID- 19135856 TI - Perceptual evaluation of severe pediatric voice disorders: rater reliability using the consensus auditory perceptual evaluation of voice. AB - The purpose of this nonrandomized prospective study was to quantify the inter- and intrarater reliability of experienced speech-language pathologist's perceptual ratings of voice in pediatric patients post-laryngotracheal reconstruction (LTR). Moderate to severe dysphonia is common in this population. Using the sentence portion of the Consensus Auditory Perceptual Evaluation-Voice (CAPE-V) rating scale, three experienced speech-language pathologists independently rated randomized voice samples of 50 participants ages 4-20 years, who had acquired or congenital airway conditions requiring at least one LTR on the six salient perceptual vocal attributes. Data collection and listening conditions were carefully controlled. Seventeen (34%) of the samples were randomly selected for rerating at a later time. Estimates of interrater reliability were strongest for perceptual ratings of breathiness (intraclass correlation coefficient [ICC]=71%), roughness (ICC=68%), pitch (ICC=68%), and overall severity (ICC=67%). Reliability was lower for ratings of loudness (ICC=57%) and strain (ICC=35%). For each rater, the intrarater reliability on all but one parameter (strain) was moderate to strong (ICC=63-93%). There was a strong interrater eliability for four of six vocal parameters rated using the CAPE-V in a population of children and adolescents with marked dysphonia. The parameter of strain, when rated by auditory sample alone and apart from the clinical context, was difficult to rate. PMID- 19135857 TI - Vocal fold bridge: a complication of a sulcus cyst surgery. AB - Vocal fold bridges are rare pathologies and mostly coincidentally discovered in direct laryngoscopy. Although the pathophysiology of formation is not well established, it is attributed as a primary pathology that accompanies sulcus and related vocal fold disorders. This paper presents a vocal fold bridge formation in a professional singer after surgical removal of a sulcus cyst formed as a sequela with typical histologic findings resembling the primary bridges. This case report addresses a very rare unpublished complication of a phonomicrosurgical procedure and implicates the evidence of trauma in the etiology. PMID- 19135858 TI - Extensive Salmonella enteritidis endocarditis involving mitral, tricuspid valves, aortic root and right ventricular wall. AB - After conducting a PubMed search, 11 articles describing a total of 12 cases of native valve Salmonella enteritidis (S. enteritidis) endocarditis were identified in the English literature. Only 13 cases of prosthetic valve endocarditis attributed to S. enteritidis have been published in the English literature. Only 1 case involving the myocardium and other valves concomitantly could be located. Transthoracic echocardiography proved inadequate to demonstrate valvular vegetations. Transesophageal echocardiography was instrumental in establishing the diagnosis of endocarditis by documenting vegetations. S. enteritidis endocarditis can cause devastating endovascular infections in immunocompromised patients. Patients who present with multiple vague symptoms with medical histories that include diabetes mellitus, immunocompromise, and prosthetic heart valves should alert clinicians to strongly consider S. enteritidis endocarditis in their differential diagnoses. Despite considerable effort, S. enteritidis endocarditis poses very high risk for morbidity and mortality. PMID- 19135859 TI - Quantile regression and restricted cubic splines are useful for exploring relationships between continuous variables. AB - OBJECTIVE: Ordinary least squares (OLS) regression, commonly called linear regression, is often used to assess, or adjust for, the relationship between a continuous independent variable and the mean of a continuous dependent variable, implicitly assuming a linear relationship between them. Linearity may not hold, however, and analyzing the mean of the dependent variable may not capture the full nature of such relationships. Our goal is to demonstrate how combined use of quantile regression and restricted cubic splines (RCS) can reveal the true nature and complexity of relationships between continuous variables. STUDY DESIGN AND SETTING: We provide a review of methodologic concepts, followed by two examples using real data sets. In the first example, we analyzed the relationship between cognition and disease duration in multiple sclerosis. In the second example, we analyzed the relationship between length of stay (LOS) and severity of illness in the intensive care unit (ICU). RESULTS: In both examples, quantile regression showed that the relationship between the variables of interest was heterogeneous. In the second example, RCS uncovered nonlinearity of the relationship between severity of illness and length of stay. CONCLUSION: Together, quantile regression and RCS are a powerful combination for exploring relationships between continuous variables. PMID- 19135860 TI - Inadequate planning and reporting of adjudication committees in clinical trials: recommendation proposal. AB - OBJECTIVES: Adjudication committees (ACs) are recommended in randomized controlled trials (RCTs) to standardize the assessment of events. We aimed to assess the reporting and functioning of ACs (synonyms: clinical event committees, endpoint committees) in clinical trials. STUDY DESIGN AND SETTING: We searched five high-impact-factor medical journals for reports of RCTs with clinical event endpoints published between January 1, 2004 and December 31, 2005. RESULTS: ACs were reported in 33.4% of the 314 reports of RCTs. ACs were reported in 29.6% of trials with low risk of misclassification (i.e., "hard" main outcome), in 47.5% of trials with medium risk of misclassification (i.e., subjective main outcome and intervention delivered in a blinded fashion) and in 31% of trials with high risk of misclassification (i.e., subjective main outcome without intervention delivered in a blinded fashion). Selected cases to be adjudicated consisted largely of events identified by site investigators (93.3%). Data provided to the AC were reported for 47.4% of ACs. CONCLUSION: Reporting of ACs is not fitted to the risk of biased misclassification. Important aspects of the functioning of ACs are insufficiently reported or raise methodological issues. We propose some recommendations for planning and reporting ACs in clinical trials. PMID- 19135861 TI - Assessment of 3D DCE-MRI of the kidneys using non-rigid image registration and segmentation of voxel time courses. AB - We have applied automated image analysis methods in the assessment of human kidney perfusion based on 3D dynamic contrast-enhanced MRI data. This approach consists of non-rigid 3D image registration of the moving kidney followed by k means clustering of the voxel time courses with split between left and right kidney. This method was applied to four data sets acquired from healthy volunteers, using 1.5 T (2 exams) and 3 T scanners (2 exams). The proposed registration method reduced motion artifacts in the image time series and improved further analysis of the DCE-MRI data. The subsequent clustering to segment the kidney compartments was in agreement with manually delineations (similarity score of 0.96) in the same motion corrected images. The resulting mean intensity time curves clearly show the successive transition of contrast agent through kidney compartments (cortex, medulla, and pelvis). The proposed method for motion correction and kidney compartment segmentation might improve the validity and usefulness of further model-based pharmacokinetic analysis of kidney function in patients. PMID- 19135862 TI - Contrast enhancement and tissues classification of breast MRI using Kalman filter based linear mixing method. AB - Much attention is currently focused on one of the newest breast examination techniques, breast MRI. Contrast-enhanced breast MRIs acquired by contrast injection have been shown to be very sensitive in the detection of breast cancer, but are also time-consuming and cause waste of medical resources. This paper therefore proposes the use of spectral signature detection technology, the Kalman filter-based linear mixing method (KFLM), which can successfully present the results as high-contrast images and classify breast MRIs into major tissues from four bands of breast MRIs. A series of experiments using phantom and real MRIs was conducted and the results compared with those of the commonly used c-means (CM) method and dynamic contrast-enhanced (DCE) breast MRIs for performance evaluation. After comparison with the CM algorithm and DCE breast MRIs, the experimental results showed that the high-contrast images generated by the spectral signature detection technology, the KFLM, were of superior quality. PMID- 19135863 TI - Malnutrition in Dutch health care: prevalence, prevention, treatment, and quality indicators. AB - OBJECTIVE: In most health care organizations there is still insufficient awareness for recognizing and treating malnourished patients. To gain more insight into nutritional care policies in Dutch health care organizations, this study investigated screening, treatment, and other quality indicators of nutritional care. METHODS: In 2007 a cross-sectional multicenter study was performed that included 20 255 patients (hospitals, n = 6021; nursing homes, n = 11 902; home care, n = 2332). A standardized questionnaire was used to study nutritional screening and treatment at the patient level and quality indicators at institutional and ward levels (e.g., malnutrition guidelines/protocols, nutritional education, and weighing policy). RESULTS: Nutritional screening was performed more often in nursing homes (60.2%) than in hospitals (40.3%) and home care (13.9%, P < 0.001). In general, one in every five patients was malnourished, and nutritional treatment was applied in fewer than 50% of all malnourished patients in nursing homes, hospitals, and home care. At ward level nursing homes focused more on the quality of nutritional care than did hospitals and home care, especially with respect to controlling the use of nutritional guidelines (54.6%, P < 0.03), weighing at admission (82.9%, P < 0.01), and mealtime ambiance (91.8%, P < 0.01). CONCLUSION: This large-scale study shows that malnutrition is still a considerable problem in one of every five patients in all participating health care settings. It furthermore demonstrates that recognizing and treating malnutrition continues to be problematic. To target the problem of malnutrition adequately, more awareness is needed of the importance of nutritional screening, appropriate treatment, and other nutritional quality indicators. PMID- 19135864 TI - Quality of life in patients with oral and oropharyngeal cancer. AB - Quality of life (QoL) is an important aspect in the clinical assessment and management of patients with cancer. The aim of the present study was to evaluate QoL at the time of diagnosis in patients with oral and oropharyngeal cancer and to establish the influence of variables such as gender, age, tumor location and tumor staging. The authors studied 149 patients with oral and oropharyngeal cancer for 2 years. QoL was measured using the EORTC QLQ-C30 and its specific modules for head and neck cancer QLQ-H&N 35. Variable deterioration of QoL was observed before therapy. The emotional domain showed the greatest alterations, while pain was the most remarkable symptom variable. QoL seems to be associated with gender (female patients obtained worse scores in most of the functional scales), age (patients < 65 years scored better), tumor location (orpharyngeal tumors showed worse prognosis) and tumor staging (early stages obtained better scores than advances ones). Many patients with oral and oropharyngeal cancer show poor QoL before initiating treatment. The present study of a homogeneous group of patients is the first carried out in Spain following the EORTC QLQ-C30 questionnaire and its results may serve for future reference. These results are similar to those obtained in populations from the north and centre of Europe. PMID- 19135866 TI - Maxillo-mandibular counter-clockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses: part II--airway changes and stability. AB - The purpose of this study was to evaluate the anatomical changes and stability of the oropharyngeal airway and head posture following TMJ reconstruction and mandibular advancement with TMJ Concepts custom-made total joint prostheses and maxillary osteotomies with counter-clockwise rotation of the maxillo-mandibular complex. All patients were operated at Baylor University Medical Center, Dallas TX, USA, by one surgeon (Wolford). The lateral cephalograms of 47 patients were analyzed to determine surgical and post-surgical changes of the oropharyngeal airway, hyoid bone and head posture. Surgery increased the narrowest retroglossal airway space 4.9 mm. Head posture showed flexure immediately after surgery ( 5.6+/-6.7 degrees) and extension long-term post surgery (1.8+/-6.7 degrees); cervical curvature showed no significant change. Surgery increased the distances between the third cervical vertebrae and the menton 11.7+/-9.1 mm and the third cervical vertebrae and hyoid 3.2+/-3.9 mm, and remained stable. The distance from the hyoid to the mandibular plane decreased during surgery (-3.8+/-5.8 mm) and after surgery (-2.5+/-5.2 mm). Maxillo-mandibular advancement with counter clockwise rotation and TMJ reconstruction with total joint prostheses produced immediate increase in oropharyngeal airway dimension, which was influenced by long-term changes in head posture but remained stable over the follow-up period. PMID- 19135865 TI - Maxillo-mandibular counter-clockwise rotation and mandibular advancement with TMJ Concepts total joint prostheses: part IV--soft tissue response. AB - The purpose of this study was to evaluate soft tissue response to maxillo mandibular counter-clockwise rotation, with TMJ reconstruction and mandibular advancement using TMJ Concepts total joint prostheses, and maxillary osteotomies in 44 females. All patients were operated at Baylor University Medical Center, Dallas TX, USA, by one surgeon (Wolford). Eighteen patients had genioplasties with either porous block hydroxyapatite or hard tissue replacement implants (Group 2) 26 had no genioplasty (Group 1). Surgically, the maxilla moved forward and upward by counter-clockwise maxillo-mandibular rotation with greater horizontal movement in Group 2. Vertically, both groups showed diversity of maxillo-mandibular mean movement. Group 1 showed a consistent 1: 0.97 ratio of hard to soft tissue advancement at pogonion; Group 2 results were less consistent, with ratios between 1: 0.84 and 1: 1.02. Horizontal changes in upper lip morphology after maxillary advancement/impaction, VY closure, and alar base cinch sutures showed greater movement in both groups, than observed in hard tissue. Counter-clockwise rotation of the maxillo-mandibular complex using TMJ Concepts total joint prostheses resulted in similar soft tissue response as previously reported for traditional maxillo-mandibular advancement without counter-clockwise rotation of the occlusal plane. The association of chin implants, in the present sample, showed higher variability of soft tissue response. PMID- 19135867 TI - Perioperative assessment of psychological state and quality of life of head and neck cancer patients undergoing surgery. AB - This study assessed psychological state and quality of life (QOL) in head and neck cancer patients during the perioperative period. Patients who had undergone primary surgery at the Department of Oral and Maxillofacial Surgery, Tokyo Medical and Dental University Hospital, Tokyo, Japan were enrolled. After obtaining informed consent, three tests were administered: the Hospital Anxiety and Depression Scale (HADS) to assess psychological state, and the Functional Assessment of Cancer Therapy General (FACT-G) and Functional Assessment of Head and Neck (FACT-H&N) to assess QOL. Japanese language versions of all tests were administered. Tests were administered 1 day before surgery, 1 week after surgery, and 1 and 6 months after discharge. Test scores were analyzed, as were relationships between psychological state, QOL and clinical factors. Anxiety was greatest before surgery, while depression was greatest immediately after surgery. QOL in the somatic and specific domains also decreased immediately after surgery. QOL in most domains improved 1 month after discharge. This study offers important information regarding perioperative psychological state and QOL in head and neck cancer patients. PMID- 19135868 TI - Provoked ictal SPECT in temporal and extratemporal drug-resistant epileptic patients: comparison of Statistical Parametric Mapping and qualitative analysis. AB - OBJECTIVE: To compare Statistical Parametric Mapping (SPM) with qualitative analysis of provoked ictal SPECT, in terms of both diagnostic yield and efficacy in identifying the epileptogenic zone of temporal and extratemporal epileptic patients. METHODS: We included in the study 27 drug-resistant epileptic patients submitted to provoked ictal SPECT and for whom both SPM and qualitative analysis were obtained. A comparison between visual SPECT analysis and SPM results was performed in terms of lateralization and localization of the ictal hyperperfusion areas. Then, hyperperfusion areas were compared to epileptogenic zones to assess whether they co-localized or not. RESULTS: In twenty patients, a co-localization was found between hyperperfusion areas detected by both qualitative and SPM analysis of provoked ictal SPECT and epileptogenic zones. A concordance between SPM and qualitative analysis in all but two patients was obtained. These findings were observed for both temporal and extratemporal epilepsies. CONCLUSIONS: The co localization between provoked ictal SPECT hyperperfusion areas and epileptogenic zones found in most patients supports the usefulness of this technique in integrating data provided by neuroimaging and electroclinical correlations to correctly define epileptogenic zone. Moreover, this preliminary work suggests that SPM as entirely objective methodology is feasible and can be accurate also after provoked ictal SPECT. PMID- 19135869 TI - Antiepileptic drug utilization in Taiwan: analysis of prescription using National Health Insurance database. AB - OBJECTIVE: The objective of this study was to analyze and evaluate antiepileptic drug (AED) utilization among adults in Taiwan. METHODS: A random sample of 167,377 patients from the National Health Insurance (NHI) reference database was used. Prescription records were retrieved for all patients prescribed AEDs during 2004. The prescribed daily dose/defined daily dose (PDD/DDD) ratio was used to assess the adequacy of AED dosing. RESULTS: Seventy-one percent (n=518) of patients used only one AED, while 29% (n=212) used more than two AEDs in 2004. For monotherapy, the most frequent regimens included carbamazepine (41.9%), followed by phenytoin (27.3%) and valproic acid (17.8%). For polytherapy, the most commonly used combination was valproic acid and carbamazepine. For adults, the mean PDD/DDD ratio for each AED used for either monotherapy or polytherapy was less than 1.00. Additionally, adult patients treated with more than one AED during 2004 in Taiwan took each drug in higher dose than patients using the same AED in monotherapy. CONCLUSION: In Taiwan antiepileptic drug therapies appear to be still dominated by the first generation drugs. The mean dosages of most antiepileptic drugs were lower than that of WHO suggested. PMID- 19135870 TI - Stereotactic radiofrequency amygdalohippocampectomy for the treatment of mesial temporal lobe epilepsy: correlation of MRI with clinical seizure outcome. AB - SUMMARY: Stereotactic radiofrequency amygdalohippocampectomy (AHE) has been reintroduced as an alternative treatment of mesial temporal lobe epilepsy. The aim of this study was to describe MRI changes after stereotactic AHE and to correlate the hippocampal and amygdalar volumes reduction with the clinical seizure outcome. Eighteen patients after stereotactic AHE were included. Volumetry was calculated from pre-operative MRI and from MRI obtained 1 year after the operation. The clinical outcome was examined 1 and 2 years after the treatment. Hippocampal volume decreased by 54+/-19%, and amygdalar volume decreased by 49+/-18%. One year after the procedure, 13 (72%) patients were classified as Engel's Class I (9 as Class IA), 4 (22%) patients as Class II and 1 (6%) patient as Class III. Two years after the operation, 14 patients (82%) were classified as Class I (7 as Class IA) and 3 patients (18%) as Class II. We found 3 surgical complications after the procedure: one small subdural hematoma, and twice a small electrode tip left in operation field (these patients were excluded from the study). In 3 patients, temporary meningeal syndrome developed. Results of radiofrequency AHE are promising. The volume reduction of target structures after AHE is significantly related to the clinical outcome. PMID- 19135871 TI - Catatonia due to a prion familial disease. PMID- 19135872 TI - Increased diffusivity in superior temporal gyrus in patients with schizophrenia: a Diffusion Tensor Imaging study. AB - BACKGROUND: Superior temporal gyrus (STG) volume reduction is one of the most consistent findings in schizophrenia. The goal of this study was to conduct the first Diffusion Tensor Imaging (DTI) study to investigate altered structural integrity in STG gray and white matter in patients with chronic schizophrenia compared with healthy controls. METHODS: Magnetic resonance imaging (MRI) and DTI were acquired in 21 male patients with schizophrenia and 22 age-, handedness-, and parental social economic status-matched male comparison subjects. After manual segmentation of gray and white matter, mean diffusivity and fractional anisotropy were measured within STG. Correlational analyses were also conducted to test possible associations between DTI and clinical measures, including positive and negative symptoms of schizophrenia. RESULTS: Compared with controls, patients demonstrated reduced volume, bilaterally, in STG gray matter but not in white matter. For DTI measures, patients showed increased mean diffusivity, bilaterally, in STG gray matter, and in left STG white matter. In addition, mean diffusivity in left STG white matter showed statistically significant correlations with auditory hallucinations and attentional impairments in patients. CONCLUSIONS: These findings suggest a disruption of tissue integrity in STG gray and white matter in schizophrenia. In addition, increased water diffusivity in left-side STG, which was associated with auditory hallucinations and attentional impairments, suggests the possibility of a disconnection among auditory/language processing regions in schizophrenia. PMID- 19135873 TI - Single nucleotide polymorphisms and the haplotype in the DEFB1 gene are associated with atopic dermatitis in a Korean population. AB - BACKGROUND: Atopic dermatitis (AD) patients have been recognized to have an increased susceptibility to cutaneous colonization and infection by bacteria, fungi and viruses. OBJECTIVE: We wanted to evaluate the associations of single nucleotide polymorphism (SNP) and the haplotype in the defensin (DEFA) and defensin (DEFB) genes, and so we performed genotyping for the SNPs in these genes in both AD patients and normal controls. METHOD: We genotyped 27 SNPs from the DEFA 4, 5 and 6 genes and the DEFB1 gene for 1089 case-control samples (631 AD patients and 458 normal controls). We analyzed the SNPs and haplotypes in each gene. RESULT: We identified that two SNPs and the haplotype CT in the DEFB1 gene are associated with AD in Koreans. The rs5743399 (-2266T/C) SNP is associated with AD, and especially with the high IgE, extrinsic type, and the rs5743409 ( 1241T/G) SNP is associated with AD. On the haplotype analysis of these two SNPs, the haplotype CT is associated with AD, and especially with the allergic, extrinsic type of AD. However, we could not find any significant associations between the SNPs in the three DEFA genes and AD. CONCLUSION: We found that the rs5743399 SNP, the rs5743409 SNP and the CT haplotype in the DEFB1 gene were significantly associated with the susceptibility to AD. We also found that rs5743399 polymorphism and the haplotype CT in this gene showed a strong association with the allergic, extrinsic type of AD. These results suggest that the DEFB1 gene has a main effect on the skin inflammation and/or skin responsiveness to any kind of allergic reaction. PMID- 19135874 TI - Antimicrobial activity of commercial Olea europaea (olive) leaf extract. AB - The aim of this research was to investigate the activity of a commercial extract derived from the leaves of Olea europaea (olive) against a wide range of microorganisms (n=122). Using agar dilution and broth microdilution techniques, olive leaf extract was found to be most active against Campylobacter jejuni, Helicobacter pylori and Staphylococcus aureus [including meticillin-resistant S. aureus (MRSA)], with minimum inhibitory concentrations (MICs) as low as 0.31 0.78% (v/v). In contrast, the extract showed little activity against all other test organisms (n=79), with MICs for most ranging from 6.25% to 50% (v/v). In conclusion, olive leaf extract was not broad-spectrum in action, showing appreciable activity only against H. pylori, C. jejuni, S. aureus and MRSA. Given this specific activity, olive leaf extract may have a role in regulating the composition of the gastric flora by selectively reducing levels of H. pylori and C. jejuni. PMID- 19135879 TI - Science diplomacy in the 21st century. AB - Science diplomacy is the use of scientific collaborations among nations to address the common problems facing 21(st) century humanity and to build constructive international partnerships. There are many ways that scientists can contribute to this process. PMID- 19135880 TI - The secret life of Smad4. AB - Smad4 is an important mediator of signaling by TGFbeta family members that is presumed to be controlled by the phosphorylation status of its partners, the receptor-regulated R-Smads. In this issue, Dupont et al. (2009) now reveal that Smad4 itself is subject to cycles of ubiquitination and deubiquitination that regulate its interactions with the R-Smads. PMID- 19135881 TI - ESCRTing membrane deformation. AB - The ESCRT-III complex mediates membrane budding away from the cytosol in endosome biogenesis, cytokinesis, and viral budding. In this issue, Saksena et al. (2009) use an elegant fluorescence-based approach to define the sequential activation, recruitment, and disassembly of ESCRT-III subunits during membrane involution in vitro. PMID- 19135882 TI - Who benefits from granulomas, mycobacteria or host? AB - By investigating host-pathogen interactions in zebrafish using intravital imaging, Davis and Ramakrishnan (2009) provide evidence that aggregates of immune cells known as granulomas, long thought to constrain mycobacterial infection, may instead facilitate its spread. PMID- 19135883 TI - When sirtuins and NF-kappaB collide. AB - Both the sirtuins and certain transcription factors of the NF-kappaB/Rel family control aging-associated changes in gene expression. Kawahara et al. (2009) now show a requirement in mice for the sirtuin SIRT6 in dampening NF-kappaB-dependent gene expression, thus unveiling a potential link between inflammation, aging, and metabolism. PMID- 19135884 TI - Are GTGs ABA's biggest fans? AB - The search for receptors for abscisic acid (ABA), a phytohormone central to the response of plants to biotic and abiotic stress, has been controversial. In this issue, Pandey et al. (2009) report the identification of two membrane proteins from Arabidopsis, GTG1 and GTG2, that bind ABA in vitro and mediate ABA responses in vivo. PMID- 19135885 TI - A new family of odorant receptors in Drosophila. AB - In the fruit fly Drosophila, not all olfactory sensory neurons express a seven transmembrane odorant receptor, suggesting that other types of odorant receptors might exist. Benton et al. (2009) now present evidence that a family of proteins related to ionotropic glutamate receptors is a previously unrecognized class of odorant receptors. PMID- 19135886 TI - MicroRNA control in the immune system: basic principles. AB - MicroRNA (miRNA) control has emerged as a critical regulatory principle in the mammalian immune system. Genetic ablation of the miRNA machinery, as well as loss or deregulation of certain individual miRNAs, severely compromises immune development and response and can lead to immune disorders like autoimmunity and cancer. Although individual miRNAs modulate protein output from hundreds of target genes, they may impact physiological processes by regulating the concentrations of just a few key cellular proteins that may be components of a single or of functionally interrelated pathways in a given cellular context. PMID- 19135887 TI - The role of the granuloma in expansion and dissemination of early tuberculous infection. AB - Granulomas, organized aggregates of immune cells, form in response to persistent stimuli and are hallmarks of tuberculosis. Tuberculous granulomas have long been considered host-protective structures formed to contain infection. However, work in zebrafish infected with Mycobacterium marinum suggests that granulomas contribute to early bacterial growth. Here we use quantitative intravital microscopy to reveal distinct steps of granuloma formation and assess their consequence for infection. Intracellular mycobacteria use the ESX-1/RD1 virulence locus to induce recruitment of new macrophages to, and their rapid movement within, nascent granulomas. This motility enables multiple arriving macrophages to efficiently find and phagocytose infected macrophages undergoing apoptosis, leading to rapid, iterative expansion of infected macrophages and thereby bacterial numbers. The primary granuloma then seeds secondary granulomas via egress of infected macrophages. Our direct observations provide insight into how pathogenic mycobacteria exploit the granuloma during the innate immune phase for local expansion and systemic dissemination. PMID- 19135888 TI - Cdk1-dependent phosphorylation of Cdc13 coordinates telomere elongation during cell-cycle progression. AB - Elongation of telomeres by telomerase replenishes the loss of terminal telomeric DNA repeats during each cell cycle. In budding yeast, Cdc13 plays an essential role in telomere length homeostasis, partly through its interactions with both the telomerase complex and the competing Stn1-Ten1 complex. Previous studies in yeast have shown that telomere elongation by telomerase is cell cycle dependent, but the mechanism underlying this dependence is unclear. In S. cerevisiae, a single cyclin-dependent kinase Cdk1 (Cdc28) coordinates the serial events required for the cell division cycle, but no Cdk1 substrate has been identified among telomerase and telomere-associated factors. Here we show that Cdk1 dependent phosphorylation of Cdc13 is essential for efficient recruitment of the yeast telomerase complex to telomeres by favoring the interaction of Cdc13 with Est1 rather than the competing Stn1-Ten1 complex. These results provide a direct mechanistic link between coordination of telomere elongation and cell-cycle progression in vivo. PMID- 19135889 TI - SIRT6 links histone H3 lysine 9 deacetylation to NF-kappaB-dependent gene expression and organismal life span. AB - Members of the sirtuin (SIRT) family of NAD-dependent deacetylases promote longevity in multiple organisms. Deficiency of mammalian SIRT6 leads to shortened life span and an aging-like phenotype in mice, but the underlying molecular mechanisms are unclear. Here we show that SIRT6 functions at chromatin to attenuate NF-kappaB signaling. SIRT6 interacts with the NF-kappaB RELA subunit and deacetylates histone H3 lysine 9 (H3K9) at NF-kappaB target gene promoters. In SIRT6-deficient cells, hyperacetylation of H3K9 at these target promoters is associated with increased RELA promoter occupancy and enhanced NF-kappaB dependent modulation of gene expression, apoptosis, and cellular senescence. Computational genomics analyses revealed increased activity of NF-kappaB-driven gene expression programs in multiple Sirt6-deficient tissues in vivo. Moreover, haploinsufficiency of RelA rescues the early lethality and degenerative syndrome of Sirt6-deficient mice. We propose that SIRT6 attenuates NF-kappaB signaling via H3K9 deacetylation at chromatin, and hyperactive NF-kappaB signaling may contribute to premature and normal aging. PMID- 19135890 TI - Posttranscriptional crossregulation between Drosha and DGCR8. AB - The Drosha-DGCR8 complex, also known as Microprocessor, is essential for microRNA (miRNA) maturation. Drosha functions as the catalytic subunit, while DGCR8 (also known as Pasha) recognizes the RNA substrate. Although the action mechanism of this complex has been intensively studied, it remains unclear how Drosha and DGCR8 are regulated and if these proteins have any additional role(s) apart from miRNA processing. Here, we report that Drosha and DGCR8 regulate each other posttranscriptionally. The Drosha-DGCR8 complex cleaves the hairpin structures embedded in the DGCR8 mRNA and thereby destabilizes the mRNA. We further find that DGCR8 stabilizes the Drosha protein via protein-protein interaction. This crossregulation between Drosha and DGCR8 may contribute to the homeostatic control of miRNA biogenesis. Furthermore, microarray analyses suggest that a number of mRNAs may be downregulated in a Microprocessor-dependent, miRNA independent manner. Our study reveals a previously unsuspected function of Microprocessor in mRNA stability control. PMID- 19135891 TI - Structural studies of a bacterial condensin complex reveal ATP-dependent disruption of intersubunit interactions. AB - Condensins are key mediators of chromosome condensation across organisms. Like other condensins, the bacterial MukBEF condensin complex consists of an SMC family protein dimer containing two ATPase head domains, MukB, and two interacting subunits, MukE and MukF. We report complete structural views of the intersubunit interactions of this condensin along with ensuing studies that reveal a role for the ATPase activity of MukB. MukE and MukF together form an elongated dimeric frame, and MukF's C-terminal winged-helix domains (C-WHDs) bind MukB heads to constitute closed ring-like structures. Surprisingly, one of the two bound C-WHDs is forced to detach upon ATP-mediated engagement of MukB heads. This detachment reaction depends on the linker segment preceding the C-WHD, and mutations on the linker restrict cell growth. Thus ATP-dependent transient disruption of the MukB-MukF interaction, which creates openings in condensin ring structures, is likely to be a critical feature of the functional mechanism of condensins. PMID- 19135892 TI - Functional reconstitution of ESCRT-III assembly and disassembly. AB - Receptor downregulation in the MVB pathway is mediated by the ESCRT complexes. ESCRT-III is composed of four protein subunits that are monomeric in the cytosol and oligomerize into a protein lattice only upon membrane binding. Recent studies have shown that the ESCRT-III protein Snf7 can form a filament by undergoing homo oligomerization. To examine the role of membrane binding and of interactions with other ESCRT components in initiating Snf7 oligomerization, we used fluorescence spectroscopy to directly detect and characterize the assembly of the Snf7 oligomer on liposomes using purified ESCRT components. The observed fluorescence changes reveal an obligatory sequence of membrane-protein and protein-protein interactions that generate the active conformation of Snf7. Also, we demonstrate that ESCRT-III assembly drives membrane deformation. Furthermore, using an in vitro disassembly assay, we directly demonstrate that Vps24 and Vps2 function as adaptors in the ATP-dependent membrane disassembly of the ESCRT-III complex by recruiting the AAA ATPase Vps4. PMID- 19135894 TI - FAM/USP9x, a deubiquitinating enzyme essential for TGFbeta signaling, controls Smad4 monoubiquitination. AB - The assembly of the Smad complex is critical for TGFbeta signaling, yet the mechanisms that inactivate or empower nuclear Smad complexes are less understood. By means of siRNA screen we identified FAM (USP9x), a deubiquitinase acting as essential and evolutionarily conserved component in TGFbeta and bone morphogenetic protein signaling. Smad4 is monoubiquitinated in lysine 519 in vivo, a modification that inhibits Smad4 by impeding association with phospho Smad2. FAM reverts this negative modification, re-empowering Smad4 function. FAM opposes the activity of Ectodermin/Tif1gamma (Ecto), a nuclear factor for which we now clarify a prominent role as Smad4 monoubiquitin ligase. Our study points to Smad4 monoubiquitination and deubiquitination as a way for cells to set their TGFbeta responsiveness: loss of FAM disables Smad4-dependent responses in several model systems, with Ecto being epistatic to FAM. This defines a regulative ubiquitination step controlling Smads that is parallel to those impinging on R Smad phosphorylation. PMID- 19135893 TI - ATPase cycle of the nonmotile kinesin NOD allows microtubule end tracking and drives chromosome movement. AB - Segregation of nonexchange chromosomes during Drosophila melanogaster meiosis requires the proper function of NOD, a nonmotile kinesin-10. We have determined the X-ray crystal structure of the NOD catalytic domain in the ADP- and AMPPNP bound states. These structures reveal an alternate conformation of the microtubule binding region as well as a nucleotide-sensitive relay of hydrogen bonds at the active site. Additionally, a cryo-electron microscopy reconstruction of the nucleotide-free microtubule-NOD complex shows an atypical binding orientation. Thermodynamic studies show that NOD binds tightly to microtubules in the nucleotide-free state, yet other nucleotide states, including AMPPNP, are weakened. Our pre-steady-state kinetic analysis demonstrates that NOD interaction with microtubules occurs slowly with weak activation of ADP product release. Upon rapid substrate binding, NOD detaches from the microtubule prior to the rate limiting step of ATP hydrolysis, which is also atypical for a kinesin. We propose a model for NOD's microtubule plus-end tracking that drives chromosome movement. PMID- 19135895 TI - Two novel GPCR-type G proteins are abscisic acid receptors in Arabidopsis. AB - In plants, G proteins modulate signaling by the stress hormone, abscisic acid (ABA). We identify and characterize two novel Arabidopsis proteins that show homology to an orphan vertebrate GPCR (GPR89) and interact with the sole Arabidopsis G protein alpha subunit, GPA1, but also have intrinsic GTP-binding and GTPase activity. We have named these proteins GPCR-type G proteins (GTG1 and GTG2). Arabidopsis mutants lacking both GTG1 and GTG2 exhibit ABA hyposensitivity. GTG1 and GTG2 bind ABA specifically. The GDP-bound form of the GTGs exhibits greater ABA binding than the GTP-bound form, the GTPase activity of the GTGs is inhibited by GPA1, and gpa1 null mutants exhibit ABA-hypersensitive phenotypes. These results predict that, unusually, it is the GDP-bound, not the GTP-bound, form of the GTGs that actively relays the signal. We propose that GTG proteins function both as a new type of G protein and as a class of membrane localized ABA receptors. PMID- 19135896 TI - Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila. AB - Ionotropic glutamate receptors (iGluRs) mediate neuronal communication at synapses throughout vertebrate and invertebrate nervous systems. We have characterized a family of iGluR-related genes in Drosophila, which we name ionotropic receptors (IRs). These receptors do not belong to the well-described kainate, AMPA, or NMDA classes of iGluRs, and they have divergent ligand-binding domains that lack their characteristic glutamate-interacting residues. IRs are expressed in a combinatorial fashion in sensory neurons that respond to many distinct odors but do not express either insect odorant receptors (ORs) or gustatory receptors (GRs). IR proteins accumulate in sensory dendrites and not at synapses. Misexpression of IRs in different olfactory neurons is sufficient to confer ectopic odor responsiveness. Together, these results lead us to propose that the IRs comprise a novel family of chemosensory receptors. Conservation of IR/iGluR-related proteins in bacteria, plants, and animals suggests that this receptor family represents an evolutionarily ancient mechanism for sensing both internal and external chemical cues. PMID- 19135897 TI - The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. AB - Mitochondria are mobile organelles and cells regulate mitochondrial movement in order to meet the changing energy needs of each cellular region. Ca(2+) signaling, which halts both anterograde and retrograde mitochondrial motion, serves as one regulatory input. Anterograde mitochondrial movement is generated by kinesin-1, which interacts with the mitochondrial protein Miro through an adaptor protein, milton. We show that kinesin is present on all axonal mitochondria, including those that are stationary or moving retrograde. We also show that the EF-hand motifs of Miro mediate Ca(2+)-dependent arrest of mitochondria and elucidate the regulatory mechanism. Rather than dissociating kinesin-1 from mitochondria, Ca(2+)-binding permits Miro to interact directly with the motor domain of kinesin-1, preventing motor/microtubule interactions. Thus, kinesin-1 switches from an active state in which it is bound to Miro only via milton, to an inactive state in which direct binding to Miro prevents its interaction with microtubules. Disrupting Ca(2+)-dependent regulation diminishes neuronal resistance to excitotoxicity. PMID- 19135899 TI - SnapShot: centriole biogenesis. PMID- 19135898 TI - Purification of proteins associated with specific genomic Loci. AB - Eukaryotic DNA is bound and interpreted by numerous protein complexes in the context of chromatin. A description of the full set of proteins that regulate specific loci is critical to understanding regulation. Here, we describe a protocol called proteomics of isolated chromatin segments (PICh) that addresses this issue. PICh uses a specific nucleic acid probe to isolate genomic DNA with its associated proteins in sufficient quantity and purity to allow identification of the bound proteins. Purification of human telomeric chromatin using PICh identified the majority of known telomeric factors and uncovered a large number of novel associations. We compared proteins found at telomeres maintained by the alternative lengthening of telomeres (ALT) pathway to proteins bound at telomeres maintained by telomerase. We identified and validated several proteins, including orphan nuclear receptors, that specifically bind to ALT telomeres, establishing PICh as a useful tool for characterizing chromatin composition. PMID- 19135900 TI - Genome-wide DNA-mapping of CD34+ cells from patients with myelodysplastic syndrome using 500K SNP arrays identifies significant regions of deletion and uniparental disomy. AB - OBJECTIVE: Identification of genomic lesions in progenitor cells of patients with myelodysplastic syndrome (MDS) could lead to the discovery of new disease specific genes and may be of prognostic value. MATERIALS AND METHODS: We carried out a genome-wide mapping of DNA from CD34+ cells of MDS patients with high resolution 500K single nucleotide polymorphism arrays and a concomitant integration with global gene expression analysis. Thirteen MDS patients were analyzed. RESULTS: Copy number and loss of heterozygosity analyses detected heterozygous deletions on chromosomes 2, 9, 13, 16, 17, and 20 ranging in size from 0.1 megabases (Mba) to 2.1 Mba. Additionally, numerous regions with significant uniparental disomy were detected. Integration of the genomic data with gene expression analysis showed that genes, which were downregulated at least 1.5-fold in regions of significant deletion and uniparental disomy were exclusively downregulated in those samples displaying the aberration. Genomics and gene expression data were confirmed by real-time polymerase chain reaction and variable number tandem repeat analysis. CONCLUSION: High-density genomic mapping of CD34+ bone marrow cells from patients with MDS identifies cryptic genetic lesions and offers new opportunities for the discovery of target genes in MDS by integration with gene expression analysis. PMID- 19135902 TI - miR-320 targets transferrin receptor 1 (CD71) and inhibits cell proliferation. AB - OBJECTIVE: MicroRNAs (miRNAs) have been implicated in complex vertebrate developmental systems, such as hematopoiesis, and may play an integral role in the development of human cancers. Based on these observations, we investigated the contribution of miRNAs to acute myelogenous leukemia cell lineage-specific differentiation. MATERIALS AND METHODS: To facilitate the identification of miRNAs and their targets relevant to leukemic cell differentiation, changes miRNA expression were analyzed in the human leukemia cell line HL-60, which historically has been utilized to study lineage-specific changes in response to the differentiation agent 12-O-tetradecanoylphorbol-13-acetate (TPA). RESULTS: Using this approach, we have identified a panel of TPA-induced miRNAs that are expressed coincident with HL-60 stereotypic morphological changes characteristic of monocytic differentiation. The transferrin receptor 1(TfR-1; CD71), whose surface expression is downregulated during TPA-mediated HL-60 cell differentiation, has been identified as a target of the TPA-induced miRNA miR 320. Cell culture experiments indicate that enforced miR-320 expression can suppress TfR-1 expression and cell proliferation. CONCLUSION: TPA induces the expression of several miRNAs in HL-60 cells, one such miRNA (miR-320) contributes to downregulation of TfR-1 surface expression characteristically seen during HL 60 monocytic differentiation. Moreover, TfR-1-targeting miRNAs, such as miR-320, may have potential as novel therapeutic agents for cancer due to their inhibitory effects on cell proliferation. PMID- 19135901 TI - Neurobeachin (NBEA) is a target of recurrent interstitial deletions at 13q13 in patients with MGUS and multiple myeloma. AB - OBJECTIVE: Chromosome 13 deletions (del[13]), detected by metaphase cytogenetics, predict poor outcomes in multiple myeloma (MM), but the gene(s) responsible have not been conclusively identified. We sought to identify tumor-suppressor genes on chromosome 13 using a novel array comparative genomic hybridization (aCGH) strategy. MATERIALS AND METHODS: We identified DNA copy number losses on chromosome 13 using genomic DNA isolated from CD138-enriched bone marrow cells (tumor) from 20 patients with MM, monoclonal gammopathy of undetermined significance, or amyloidosis. We used matched skin biopsy (germline) genomic DNA to control for copy number polymorphisms and a novel aCGH array dedicated to chromosome 13 to map somatic DNA gains and losses at ultra-high resolution (>385,000 probes; median probe spacing 60 bp). We analyzed microarray expression data from an additional 262 patient samples both with and without del[13]. RESULTS: Two distinct minimally deleted regions at 13q14 and 13q13 were defined that affected the RB1 and NBEA genes, respectively. RB1 is a canonical tumor suppressor previously implicated in MM. NBEA is implicated in membrane trafficking in neurons, protein kinase A binding, and has no known role in cancer. Noncoding RNAs on chromosome 13 were not affected by interstitial deletions. Both the RB1 and NBEA genes were deleted in 40% of cases (8 of 20; 5 patients with del[13] detected by traditional methods and 3 patients with interstitial deletions detected by aCGH). Forty-one additional MM patient samples were used for complete exonic sequencing of RB1, but no somatic mutations were found. Along with RB1, NBEA gene expression was significantly reduced in cases with del[13]. CONCLUSIONS: The NBEA gene at 13q13, and its expression are frequently disrupted in MM. Additional studies are warranted to evaluate the role of NBEA as a novel candidate tumor-suppressor gene. PMID- 19135903 TI - Quitting by gradual smoking reduction using nicotine gum: a randomized controlled trial. AB - BACKGROUND: Many smokers express a desire to quit smoking by gradually reducing the number of cigarettes they smoke until they stop completely. This study tested the efficacy of nicotine gum in facilitating cessation through gradual reduction. DESIGN: This was a multi-center, placebo-controlled, double-blind RCT of 2- and 4 mg nicotine gum versus placebo. SETTING/PARTICIPANTS: 3297 smokers who were interested in quitting gradually. INTERVENTION: Subjects were instructed to gradually reduce their smoking while increasing their gum use over the course of up to 8 weeks. Once they had achieved initial abstinence (no smoking for 24 hours), gum was to be used in accordance with the current FDA-approved directions for cessation. The study was conducted under over-the-counter conditions, with no counseling provided. Continuous abstinence was assessed after 28 days and 6 months. Secondary measures of smoking reduction were also assessed. Analyses were conducted in 1999-2000 and 2007-2008. MAIN OUTCOME MEASURES: Smokers on active gum were significantly more likely to achieve initial cessation (2 mg: OR=1.42; 4 mg: OR=1.90); 28-day continuous abstinence (2 mg: OR=2.01; 4 mg: OR=4.66); and continuous abstinence at 6 months (2 mg: OR=1.80; 4 mg: OR=5.96). During the reduction phase, active gum aided smoking reduction, and participants who reduced their smoking were more likely to achieve abstinence. CONCLUSIONS: These findings demonstrate that smokers who wish to quit smoking by gradual reduction can increase their success by using nicotine gum to facilitate reduction and cessation. PMID- 19135904 TI - Sedentariness, small-screen recreation, and fitness in youth. AB - BACKGROUND: There are concerns that sedentariness among young people has increased and that this may be detrimental to their health. The purpose of this study was to examine the association between sedentary activities, including small-screen recreation (SSR: watching TV/DVDs/videos, recreational computer use) and cardiorespiratory endurance (CRE) in children aged 11-15 years. METHODS: A cross-sectional representative population survey was taken of New South Wales (Australia) school students in Grades 6, 8, and 10 (N=2750) in 2004. Sedentary activities and SSR were measured by a self-report questionnaire. CRE was determined by the 20-meter multi-stage shuttle run test. The optimal cut point for time spent on SSR was determined by receiver operating characteristic analysis. RESULTS: Time spent in sedentary activities was inversely associated with CRE among Grade 8 students (p=0.01) and Grade 10 girls (p=0.03). CRE was lower among Grade 8 students (p<0.001) and Grade 10 girls (p<0.001) who spent >or=2 hours/day on SSR compared with students who spent <2 hours/day. The 2 hour/day cut point for SSR had high sensitivity (boys: 84%, [95% CI=79%, 87%]; girls: 79% [95% CI=74%, 84%]) and low specificity (boys: 28% [95% CI=24%, 32%]; girls: 42% [95% CI=38%, 46%]). CONCLUSIONS: Cardiorespiratory endurance did not differ consistently across quintiles of sedentariness and SSR among boys; however, among girls there was a consistent inverse association. If confirmed in prospective studies, these findings have important implications for risk of chronic disease. PMID- 19135905 TI - Move to Improve: a randomized workplace trial to increase physical activity. AB - BACKGROUND: Workplaces are important settings for interventions to increase physical activity, but effects have been modest. PURPOSE: To evaluate the efficacy of Move to Improve, a social-ecologic intervention delivered at the workplace to increase leisure-time physical activity. METHODS: A group-randomized 12-week intervention consisting of organizational action and personal and team goal-setting was implemented in Fall 2005, with a multi-racial/ethnic sample of 1442 employees at 16 worksites of The Home Depot, Inc. Change in physical activity was analyzed in Fall 2007 using latent growth modeling and latent transition analysis. RESULTS: Participants in the intervention had greater increases in moderate and vigorous physical activity and walking compared to participants in a health education control condition. The proportion of participants that met the Healthy People 2010 recommendation for regular participation in either moderate or vigorous physical activity remained near 25% at control sites during the study but increased to 51% at intervention sites. During the last 6 weeks of the study, intervention participants exceeded 300 weekly minutes of self-reported moderate-to-vigorous physical activity and 9000 daily pedometer steps. CONCLUSIONS: The results support the feasibility and efficacy of the Move to Improve intervention and the role of goal-setting for attaining increased physical activity levels. PMID- 19135906 TI - Reducing low birth weight through home visitation: a randomized controlled trial. AB - BACKGROUND: Poor birth outcomes increase the risk of infant mortality and morbidity, developmental delays, and child maltreatment. This study assessed the effectiveness of a prenatal home-visitation program in reducing adverse birth outcomes among socially disadvantaged pregnant women and adolescents. DESIGN: As part of a larger RCT, this study examined the effects of home-visitation services on low birth weight (LBW) deliveries. SETTING/PARTICIPANTS: Pregnant women and adolescents eligible for Healthy Families New York (HFNY) were recruited in three communities. Eligibility was based on socioeconomic factors such as poverty, teen pregnancy, and the risk of child maltreatment. Two thirds of the participants were black or Hispanic, and 90% were unmarried. INTERVENTION: Pregnant women and adolescents were randomized to either an intervention group that received bi weekly home-visitation services (n=236) or to a control group (n=265). Home visitors encouraged healthy prenatal behavior, offered social support, and provided a linkage to medical and other community services. Services were tailored to individual needs. MAIN OUTCOME MEASURE: An LBW of <2500 grams on birth certificate files. Baseline and birth interviews were conducted from 2000 to 2002, and birth records were collected in 2007. Analyses were done from 2007 to 2008. RESULTS: The risk of delivering an LBW baby was significantly lower for the HFNY group (5.1%) than for the control group (9.8%; AOR=0.43; 95% CI=0.21, 0.89). The risk was further reduced for mothers who were exposed to HFNY at a gestational age of or=90% African American (or subgroup analysis), adults (>or=18 yrs), and were published in English; final article data abstraction occurred from October 2007 through February 2008. EVIDENCE SYNTHESIS: A total of 2797 titles were identified from the initial search, and 90 were deemed eligible for abstract review. Of these, 17 articles were eligible for full review and ten met all eligibility criteria. The median sample size was 761 (234 to 10,623), and half of the articles included only African Americans. Light traffic, the presence of sidewalks, and safety from crime were more often positively associated with physical activity, although associations were not consistent (OR range = 0.53 2.43). Additionally, perceived barriers to physical activity were associated with obesity. The presence of supermarkets and specialty stores was consistently positively associated with meeting fruit and vegetable guidelines. CONCLUSIONS: With relatively few studies in the literature focused on African Americans, more research is needed to draw conclusions on features of the built environment that are associated with physical activity, diet, and obesity. PMID- 19135909 TI - Health professionals' roles in animal agriculture, climate change, and human health. AB - What we eat is rapidly becoming an issue of global concern. With food shortages, the rise in chronic disease, and global warming, the impact of our dietary choices seems more relevant today than ever. Globally, a transition is taking place toward greater consumption of foods of animal origin, in lieu of plant based diets. With this transition comes intensification of animal agriculture that in turn is associated with the emergence of zoonotic infectious diseases, environmental degradation, and the epidemics of chronic disease and obesity. Health professionals should be aware of these trends and consider them as they promote healthier and more environmentally-sustainable diets. PMID- 19135910 TI - Total laparoscopic repair of abdominal aortic aneurysm with short proximal necks. AB - With the development of endovascular aneurysm repair, abdominal aortic aneurysms with short infrarenal necks (< or =10 mm, AAASN) are considered juxtarenal aneurysms. Minimally invasive treatment consists of hybrid procedures or fenestrated endografts. We present our experience with direct aortic repair for AAASN performed via a total laparoscopic approach. Data are expressed as median values with extremes. From February 2002 to December 2007, 32 patients had total laparoscopic AAASN repair. Length of the infrarenal aortic neck was 5 mm (0-10). Median age of the 29 men and three women was 70 years (range 50-84). Nine patients presented with preoperative grade 1 renal insufficiency (28.1%). The procedure was totally laparoscopic in 30 patients (93.7%). Aortic approaches included left retrorenal (n = 24) and transperitoneal left retrocolic (n = 8) exposures. Median operative and clamping times were 270 (range 215-410) and 83 (range 36-147) min, respectively. Aortic clamping was suprarenal in 14 cases (43.7%), with suprarenal clamping time of 24 min (range 9-37). Median blood loss was 850 mL (range 215-2,400). Thirty-day mortality was 3.1% (one patient died from myocardial infarction). Two patients presented with severe systemic complications (6.4%, postoperative coagulopathy with hemorrhagic syndrome, pneumopathy). Seventeen patients developed mild or moderate systemic nonlethal complications (53.1%): transient renal insufficiencies (n = 12), grade 1 ischemic colitis (n = 1), surrenal insufficiency (n = 1), myocardial ischemia (n = 1), and cardiac arythmia (n = 2). One patient was reoperated for an intestinal obstruction. Liquid diet was reintroduced after 1 day (range 1-13). Most patients were ambulatory by day 3 (range 2-17). Median lengths of stay were 48 hr (range 12-552) in the intensive care unit and 10 days (range 4-37) in the hospital. With a median follow-up of 27 months (range 1-50), 28 patients are alive, with complete recovery without graft anomalies. Three patients died, from pneumopathy (n = 1) and carcinoma (n = 2), respectively, at 29, 19, and 44 months' follow-up. Two patients presented stable juxta-renal aortic dilation <35 mm. Total laparoscopic juxtarenal AAA repair is feasible and worthwhile for patients with AAASN. Short- and midterm results match well with those of open surgery. Total laparoscopic repair in AAASN reduces the trauma of extensive surgical approaches. Based on these encouraging early results, we elected to perform laparoscopy whenever possible in good surgical risk patients with AASN. PMID- 19135911 TI - Treatment of superior mesenteric artery portal vein fistula with balloon expandable stent graft. AB - Visceral artery to portal vein arteriovenous fistulas are rare and difficult to treat. Covered stents have made treatment of pseudoaneurysms and arteriovenous fistulas feasible utilizing minimally invasive techniques. We present a case of a 46-year-old male with a remote history of an exploratory laparotomy after a motorcycle accident who presented with abdominal pain, malaise, and jaundice. A computed tomographic scan revealed a superior mesenteric artery to portal vein arteriovenous fistula. A large arteriovenous fistula was confirmed by arteriography. A balloon-expandable stent graft was placed across the arteriovenous fistula in the superior mesenteric artery. Postprocedure, the patient's abdominal pain resolved and his bilirubin decreased from 2.9 to 0.4. Endovascular repair of a superior mesenteric to portal arteriovenous fistula utilizing a stent graft is feasible and minimally invasive. PMID- 19135912 TI - Staphylococcal Infections. Preface. PMID- 19135913 TI - Staphylococcal infections: a historical perspective. AB - Staphylococcus aureus is an unusually successful and adaptive human pathogen that can cause epidemics of invasive disease despite its frequent carriage as a commensal. Over the past 100 years and more, S aureus has caused cycles of outbreaks in hospitals and the community and has developed resistance to every antibiotic used against it, yet the exact mechanisms leading to epidemics of virulent disease are not fully understood. Approaches such as bacterial interference have been effective in interrupting outbreaks, but to better prevent staphylococcal disease, we will need to be vigilant about environmental factors that facilitate its spread. Even more importantly, we need to understand more about the mechanisms that lead to its virulence and transmission. With such information, it may be possible to develop a vaccine that will prevent endemic and epidemic staphylococcal disease. PMID- 19135915 TI - Staphylococcus aureus: a community pathogen. AB - Staphylococcus aureus is a common human pathogen. S aureus infections most commonly clinically manifest as skin infections. There has been much interest in S aureus infections in the community over the past decade because of the rise of community-associated methicillin-resistant S aureus (CA-MRSA) infections, which have emerged globally over a relatively short period of time. In contrast to health care-associated methicillin resistant S aureus (HA-MRSA), circulating strains of CA-MRSA have characteristic pathogenesis, strain characteristics, epidemiology, and clinical manifestations that are distinct from HA-MRSA. In fact, CA-MRSA probably behaves more like community-associated methicillin sensitive S aureus (MSSA). This article reviews current knowledge of the epidemiology and clinical manifestations of community-associated S aureus and CA MRSA infections. PMID- 19135914 TI - Host defense and pathogenesis in Staphylococcus aureus infections. AB - Staphylococcus aureus is the most abundant cause of bacterial infections in the United States. As such, the pathogen has devised means to circumvent destruction by the innate immune system. Neutrophils are a critical component of innate immunity and the primary cellular defense against S aureus infections. This article reviews human neutrophil function in the context of S aureus virulence mechanisms and provides an overview of community-associated methicillin-resistant S aureus pathogenicity. PMID- 19135916 TI - Staphylococcal surgical site infections. AB - Staphylococcus aureus is the leading cause of surgical site infections (SSI) in the United States. In particular, SSI caused by methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a devastating complication, leading to increased mortality rates, increased length of hospitalization, and increased costs. Proven strategies for prevention of SSI caused by S aureus include addressing modifiable risk factors and correct choice and timing of antimicrobial prophylaxis. Other strategies, including decolonization and the use of vancomycin, remain controversial. PMID- 19135917 TI - Coagulase-negative staphylococcal infections. AB - Coagulase-negative staphylococci (CNS) are differentiated from the closely related but more virulent Staphylococcus aureus by their inability to produce free coagulase. Currently, there are over 40 recognized species of CNS. These organisms typically reside on healthy human skin and mucus membranes, rarely cause disease, and are most frequently encountered by clinicians as contaminants of microbiological cultures. However, CNS have been increasingly recognized to cause clinically significant infections. The conversion of the CNS from symbiont to human pathogen has been a direct reflection of the use of indwelling medical devices. This article deals with the clinical syndromes, epidemiology, prevention, and management of infections caused by this unique group of organisms. PMID- 19135918 TI - Antistaphylococcal agents. AB - These are interesting times in the treatment of infections caused by Staphylococcus aureus, with shifting epidemiology of antibiotic resistance; changing prevalence of clinical syndromes (probably reflecting changes in virulence of circulating strains); and the recent availability of a variety of new agents with activity against multidrug-resistant gram-positive cocci. The abundance of riches in new drugs for the multidrug-resistant gram-positive space is timely, and these agents show great potential, but as yet have incompletely tested durability and comparative efficacy. This article reviews the advantages and disadvantages of a variety of antistaphylococcal agents by providing basic information including mechanism of action; mechanisms of resistance; clinical use (including dosing for and data supporting common indications); drug toxicities; and major drug interactions. PMID- 19135919 TI - Staphylococcus aureus decolonization as a prevention strategy. AB - Decolonization may be defined as treatment to eradicate Staphylococcus aureus or methicillin-resistant S aureus (MRSA) carriage. Potential benefits of decolonization include decreased risk of subsequent staphylococcal infection and prevention of staphylococcal transmission to reduce endemic rates of infection or manage outbreaks. This article reviews available data regarding various proposed treatment regimens for eradicating staphylococcal carriage and the effectiveness of decolonization for infection prevention and as an infection control measure. PMID- 19135920 TI - Staphylococcal vaccines and immunotherapies. AB - Staphylococcus aureus is an important pathogen in the hospital and in the community, and it is increasingly resistant to multiple antibiotics. A nonantimicrobial approach to controlling S aureus is needed. The most extensively tested vaccine against S aureus, which is a capsular polysaccharide-based vaccine known as StaphVAX, showed promise in an initial phase 3 trial, but was found to be ineffective in a confirmatory trial, leading to its development being halted. Likewise, a human IgG preparation known as INH-A21 (Veronate) with elevated levels of antibodies to the staphylococcal surface adhesins ClfA and SdrG made it into phase 3 testing, where it failed to show a clinical benefit. Several novel antigens are being tested for potential inclusion in a staphylococcal vaccine, including cell wall-anchored adhesin proteins and exotoxins. Given the multiple and sometimes redundant virulence factors of S aureus that enable it to be such a crafty pathogen, if a vaccine is to prove effective, it will have to be multicomponent, incorporating several surface proteins, toxoids, and surface polysaccharides. PMID- 19135921 TI - Heterogeneity of magnetic resonance imaging in Leigh syndrome with SURF1 gene 604G-->C mutation. AB - OBJECTIVE: To identify the magnetic resonance (MR) features of a group of pediatric patients with Leigh syndrome (LS) caused by SURF1 gene 604G-->C mutation. MATERIALS AND METHODS: Eight cases with definite diagnosis of SURF1 gene 604G-->C mutation in our hospital were reviewed. Most cases presented typical symptoms in their infancy or childhood, with psychomotor regression, hypotonia, or eye movement disturbances. They all underwent cranial MR examinations after the onset. Their brain images were reviewed by an experienced neuroradiologist to determine the abnormalities. RESULTS: The data of our group showed heterogeneous neuroradiological findings: involvement of the brain stem and subthalamic nuclei was found in only three cases; basal ganglia abnormalities were detected in two cases, whereas demyelination was demonstrated in four cases; and brain atrophy existed invariably in the group. CONCLUSION: The MR presentation in LS patients with SURF1 gene 604G-->C mutation is variable. Maybe it is not appropriate to correlate a specific gene mutation with a homogenous radiological pattern. PMID- 19135922 TI - Reduced compression mammography to reduce breast pain. AB - This study evaluated whether reduced compression mammography to relieve breast tenderness is feasible. Women can better tolerate a compression force of approximately 90 N in mammography. The breast thickness increased approximately 3 mm when the compression force was reduced to 90 N, and although the radiation dose increased approximately 20%, the image quality was identical to that with standard compression. Many patients experience breast pain with a compression force of 120 N. Reduced compression force mammography is acceptable in women whose breasts are particularly sensitive. PMID- 19135923 TI - Appearances of pulmonary focal lesions at 0.5-mm slice thickness computed tomography: comparison with 1-mm slice thickness computed tomography. AB - AIM: This study aimed to evaluate the validity of 0.5-mm thin-section computed tomography (CT) for the assessment of pulmonary nodular lesion in comparison with 1-mm CT. MATERIALS AND METHODS: A total of 38 focal lesions from 30 patients, which were scanned with 0.5- and 1.0-mm collimation, were evaluated regarding the extent of ground-glass opacity (GGO) and well-defined margin, and the presence of pleural indentation, spicula, and internal air density. The frequency of each finding was statistically compared between 0.5- and 1-mm CT using the McNemar test. RESULTS: No statistically significant difference was observed between 0.5- and 1-mm CT for each finding. CONCLUSION: The use of 0.5-mm CT is not justified if the original collimation of multi-detector row CT is near 1 mm. PMID- 19135924 TI - Value of multiplanar reconstruction in MSCT in demonstrating the relationship between solitary pulmonary nodule and bronchus. AB - In this report, we studied the value of the solitary pulmonary nodule (SPN) bronchus relationship in determining the nature of SPN by multiplanar reconstruction (MPR) in multislice spiral computed tomography (MSCT). One hundred forty-eight SPN cases were enrolled. CT was performed in all cases using MSCT. Images were then transferred to a processing workstation for MPR. The results showed that MPR is a valuable tool for visualizing the SPN-bronchus relationship and that the SPN-bronchus relationship is useful in determining the nature and the degree of differentiation of SPN. PMID- 19135925 TI - Veins around the esophagus and the stomach: do their calibrations provide a diagnostic clue for portal hypertensive gastropathy? AB - OBJECTIVE: We analyze whether calibrations of esophagogastric veins can be used as an indirect sign for the presence of portal hypertensive gastropathy (PHG). PATIENTS AND METHODS: The diameters of the left gastric, paraesophageal, and azygos veins were measured on magnetic resonance (MR) portograms of 57 patients with portal hypertension. The mean diameters of the above mentioned veins in patients with and without PHG were compared with Mann-Whitney U test. The diametral associations between the left gastric and azygos veins were assessed with Fisher's exact test in respect to the presence of PHG. RESULTS: In patients with PHG, the mean diameters of the left gastric, paraesophageal, and azygos veins were not statistically different from those in patients without PHG (P>.05). There was no significant difference in the diametral relationship between the left gastric and azygos veins in patients with and without PHG. CONCLUSION: The calibrations of the veins around the esophagus and the stomach do not provide any clue about the presence of PHG. PMID- 19135926 TI - Cinematographic techniques in the diagnostics of intestinal diseases using MRT enteroclysma. AB - PURPOSE: We investigated whether an additional cine sequence in MR enteroclysis examination is practicable and would provide additional findings in the diagnostics of patients with abdominal diseases. MATERIALS AND METHODS: Seventeen consecutively presenting patients with known or presumed abdominal disease (known or presumed intestinal illness; age range, 21-57 years; mean age, 34.1 years; male:female ratio, 4:13) were retrospectively studied. All patients underwent MRI enteroclysis at 1.5 T subsequent to conventional ileocolonoscopy, which was used as reference standard. A gastroenterologist and a radiologist evaluated the MRI examinations together to see whether the additional cine sequences would provide any additional findings. RESULTS: The cine sequences provided relevant diagnostic information in at least 11 of 17 patients (60.6%). In five patients, we were able to determine a stenosis as scarred; in two patients, normal peristalsis of the bowel was demonstrated, showing that the stenosis had not been scarred. CONCLUSION: This study provides strong evidence that the implementation of cine sequences in MRI enteroclysis examination provides additional findings with clinical relevance, especially for distinguishing scarred and functional stenoses. PMID- 19135927 TI - Why can we not see a normal appendix on CT? An evaluation of the factors influencing nonvisualization of a normal appendix by 64-slice MDCT. AB - PURPOSE: The aim of this study was to identify the factors that prevent the visualization of a normal appendix by 64-slice MDCT. MATERIALS AND METHODS: Thirty-nine consecutive patients with a normal appendix not visualized during routine abdominal CT and without a history of abdominal surgery were selected for this study. In addition, 100 consecutive patients with a clearly visualized normal appendix by CT were selected as controls. Two radiologists, by consensus, evaluated MDCT images for cecum level, pericecal fat content, presence of small bowel dilatation, presence of pericecal fluid, presence of cecal wall thickening, and identification of the ileocecal valve. RESULTS: Patients with a nonvisualized normal appendix had a significantly lower cecum level, minimal pericecal fat, and a higher amount of pericecal fluid and were less likely to have an identified ileocecal valve (46.2%, 79.5%, 17.9%, and 76.9%, respectively) than patients with a clearly visualized appendix (18%, 9%, 4%, and 100%, respectively) (all P<.01). The presence of small bowel dilatation and cecal wall thickening was not statistically significant (7.7% vs. 8% and 7.7% vs. 9%, respectively) (all P>.01). CONCLUSION: The factors that influence the nonvisualization of a normal appendix on MDCT images are as follows: a low cecum level, minimal pericecal fat, presence of pericecal fluid, and nonidentification of the ileocecal valve. PMID- 19135928 TI - Periportal oedema of the liver: correlation with clinical and paraclinical parameters in polytraumatic patients. AB - PURPOSE: To clarify the occurrence of periportal edema in polytraumatic patients. MATERIALS AND METHODS: Retrospective analysis of computed tomography (CT) scans from 74 polytraumatic patients (12 females, 62 males; 14-88 years old, median 32 years) performed shortly after the trauma. Periportal oedema was found in 22 patients. The existence and extent of the periportal oedema were studied with regard to the injury type, sex, weight and age, heart frequency, arterial blood pressure, as well as the lactate, pH value, and base excess (BE) directly after the admission of the patient to the intensive care unit. RESULTS: There is a relevant statistical correlation between the existence of periportal oedema and the abdominal trauma (P<.0001), independent of the type of abdominal injury. No relevant correlation between periportal oedema and existence of liver rupture, liver haematoma, other abdominal organ injury, abdominal vessel injury, fracture in the skeletal system, or intracerebral bleeding was found. Periportal oedema occurs more commonly in females than in males, more in light weight patients than in the others, and more frequent in young patients than in older ones. There is no correlation with arterial blood pressure, heart rate, pH value, lactate, and BE. CONCLUSION: The existence of periportal oedema is one sign of abdominal trauma and is independent of liver injury. Although it correlates with the sex, weight, and age of the patient, there is no correlation with arterial blood pressure, heart rate, pH value, lactate, and BE. PMID- 19135929 TI - Frequency and etiology of solitary hot spots in the pelvis at whole-body positron emission tomography/computed tomography imaging. AB - PURPOSE: To determine the frequency and etiology of a single hypermetabolic focus within the pelvis with no other areas of increased 18-fluorodeoxyglucose (FDG) uptake in the reminder of the whole body in an oncological population. METHOD AND MATERIALS: We retrospectively examined the first 700 whole-body PET/CT scans performed at our institution for baseline staging or follow-up of cancer and identified all patients with a solitary focus of increased FDG uptake in the pelvis. All available medical records and imaging findings in these patients were reviewed in order to determine the etiology of increased FDG uptake. RESULTS: Eight (1.1%) of the 700 patients had a solitary hot spot in the pelvis at positron emission tomography (PET)/computed tomography (CT) imaging, consisting of seven of 380 women and one of 320 men. In the seven women, increased FDG uptake was due to physiological endometrial uptake (n=2), leiomyoma (n=1), corpus luteum cyst (n=1), physiological ovarian uptake (n=1), urinary leak (n=1), and nonspecific colitis (n=1). In the man, uptake was due to recurrent rectosigmoid adenocarcinoma. None of the 700 patients was found to have metastatic disease in the pelvis. CONCLUSION: Isolated pelvic hot spots at PET/CT imaging in an oncological population are not common and usually benign; physiological endometrial or ovarian uptake is the single commonest cause. PMID- 19135930 TI - The role of single-photon emission computed tomography/computed tomography for precise localization of metastases in patients with differentiated thyroid cancer. AB - PURPOSE: It is very important in the management of patients with differentiated thyroid cancer (DTC) to precisely localize the foci of I-131 uptake, but it is difficult because of a lack of anatomic landmarks. The purpose of this study was to investigate the added value of I-131 single-photon emission computed tomography (SPECT)/computed tomography (CT) fusion imaging using a hybrid system in patients with DTC. METHODS: Ninety-four patients with DTC underwent I-131 SPECT/CT using a hybrid tomography consisting of a dual-head variable-angle gamma camera and a low-dose X-ray tube. Results were compared with I-131 whole-body scan (WBS). SPECT/CT was performed 5-7 days after administration of a therapeutic dose of I-131. Fusion images were constructed by combining the digital CT and SPECT images on a computer workstation. RESULTS: Compared with I-131 WBS, SPECT/CT imaging had improved the precise localization in 21% (20/94) of patients. In addition, SPECT/CT provided additional clinical data in 12 of the patients examined (12/94) and also caused physicians to reconsider the (131)I therapeutic approach in 22 patients. CONCLUSION: The results of the current study indicate that the addition of I-131 SPECT/CT to WBS can improve the localization of metastases in patients with DTC. It may also detect metastases missed by WBS and adjust the therapy plan. PMID- 19135931 TI - Recurrent liver hydatid disease: when does it become symptomatic and how does one diagnose it? AB - BACKGROUND: Echinococcosis is a zoonotic disease that mainly occurs in sheep grazing areas. Recurrence of the disease and its diagnosis are relatively new areas of investigation due to the limited number of cases. The aim of this study was to evaluate the diagnosis of the symptomatic recurrent liver hydatid disease and the efficacy of abdominal ultrasonography (US). MATERIALS AND METHODS: Between 1988 and 2006, 412 patients with hydatid cyst of the liver were operated at Erciyes University Medical Faculty. We have considered a growing cyst at the original operative site or at the neighboring hepatic tissue as recurrence and investigated 38 of them for the recurrence of the disease (9.2%). RESULTS: The recurrence usually occurred after 2 years. The majority of the cases were Types 2 and 3 (24 cases; 8 were Type 4 and 6 were Type 1) according to Gharbi classification. In 35 patients with recurrent disease approved after surgical exploration, US was successful in preoperative diagnosis (100%). Overall, the recurrence was demonstrated correctly by means of US in 35 patients (92.1%), and 23 of 26 patients (88.4%) were correctly diagnosed with the use of computed tomography. In our study, in the first 2 years after the operation, the success rate of US examination was 72.7%. The success rate of US rose up to 100% when the frequency of recurrence is highest. CONCLUSION: Beyond 2 years after surgery, US examination alone might be enough for the diagnosis of symptomatic recurrent liver hydatid disease in patients with the suspicion of recurrence. Postoperative early US profile and annual US examination must be performed for at least 5 years to prevent misinterpretation in doubtful cases. PMID- 19135932 TI - Replaced right hepatic artery coursing ventral to the portal vein: demonstration with 64-row CT and 3D reformatted images. AB - We report one case of replaced right hepatic artery coursing ventral to the portal vein. This variation, which was demonstrated by 64-row CT angiography and three-dimensional reformatted images, has never been reported. Surgeons involved in hepatic and biliopancreatic procedures should be aware of this possible variation, which has important surgical implications. PMID- 19135933 TI - Synchronous bilateral mucinous carcinoma of the breast: case report. AB - A mucinous carcinoma of the breast is a well-differentiated rare histological type of invasive ductal carcinoma, having a lower frequency of metastasis to an axillary lymph node and a better survival rate. Bilateral breast cancer has an overall incidence of 4% to 20% in patients with primary operable breast cancer. Few reports exist in the clinical literature characterizing a synchronous bilateral mucinous carcinoma of the breast. We report the characteristic imaging findings of a bilateral mucinous carcinoma of the breast. PMID- 19135934 TI - Paratracheal air collection in a trauma patient: a case report. AB - The presence of a paratracheal air collection in the setting of acute trauma may indicate a wide array of etiologies. When a paratracheal air collection is found in a trauma patient, the possibility of tracheal or esophageal rupture must be considered. Tracheal diverticulae are most frequently found incidentally during autopsy, with an incidence of approximately 1%. We present a case of an incidental tracheal diverticulum in a patient with a recent history of motor vehicle accident with chest and neck trauma. PMID- 19135935 TI - Dedifferentiated liposarcoma of the retroperitoneum with osteosarcomatous component: report of two cases. AB - We describe two cases of ossified soft tissue tumors of the retroperitoneum. Computed tomographic and magnetic resonance imaging both revealed retroperitoneal masses consisting of two components -- densely ossified and lipid-rich components. In one case, a 50-year-old man, a histological diagnosis of dedifferentiated liposarcoma with osteosarcoma was made based on the needle biopsy of the two components. In another case, a 54-year-old man, surgical resection of the complex perirenal mass was performed and the same diagnosis was made. Although an ossified component represent high-grade lesion, the fatty component is an important clue to the diagnosis of dedifferentiated liposarcoma. The imaging features may be similar to those of malignant mesenchymoma, which is not a currently used term. PMID- 19135937 TI - A non-leaky Artemis-deficient mouse that accurately models the human severe combined immune deficiency phenotype, including resistance to hematopoietic stem cell transplantation. AB - Two Artemis-deficient (mArt(-/-)) mouse models, generated independently on 129/SvJ backgrounds, have the expected T(-)B(-)NK(+) severe combined immune deficiency (SCID) phenotype but fail to mimic the human disease because of CD4(+) T cell leakiness. Moreover, immune reconstitution after hematopoietic stem cell transplantation is achieved more readily in these leaky mouse models than in Artemis-deficient humans. To develop a more clinically relevant animal model, we backcrossed the mArt(-/-) mutation onto the C57Bl/6 (B6) background (99.9%), which resulted in virtually no CD4(+) T cell leakiness compared with 129/SvJ mArt(+/-) mice (0.3% +/- 0.25% vs 19.5% +/- 15.1%, P < .001). The nonleaky mouse also was uniquely resistant to engraftment using allogeneic mismatched hematopoietic stem cells, comparable to what is seen in human Artemis deficiency. The genetic background also influenced Artemis-associated radiation sensitivity, with differing degrees of x-ray hypersensitivity evident in 129/SvJ and B6 backgrounds with both the mArt(-/-) and mArt(+/-) genotypes. Our results indicate that immunogenic and DNA repair phenotypes associated with Artemis deficiency are significantly altered by genetic background, which has important implications for the diagnosis and treatment of SCID. Moreover, the B6 mArt(-/-) mouse provides a more accurate model for the human disease and a more appropriate system for studying human Artemis deficiency and for developing improved transplantation and gene therapy regimens for the treatment of children with SCID. PMID- 19135938 TI - Changes in quality-of-life and psychosocial adjustment among multiple myeloma patients treated with high-dose melphalan and autologous stem cell transplantation. AB - High-dose melphalan and autologous hematopoietic stem cell transplantation (HSCT) is a standard treatment for myeloma, but very little is known about the psychosocial or quality-of-life difficulties that these patients encounter during treatment. Data regarding older patients is particularly scarce. Using a prospective design, this investigation evaluated 94 patients at stem cell collection and again after high-dose therapy and transplantation. Outcomes included quality-of-life (FACT-BMT) and psychosocial adjustment (ie, Brief Symptom Inventory, Impact of Events Scale, and Satisfaction with Life Scale). Findings were compared with age- and sex-adjusted population norms and with transplantation patient norms. At stem cell collection, physical deficits were common, with most patients scoring 1 standard deviation below population norms for physical well-being (70.2%) and functional well-being (57.5%), and many reporting at least moderate fatigue (94.7%) and pain (39.4%). Clinically meaningful levels of anxiety (39.4%), depression (40.4%), and cancer-related distress (37.0%) were evident in a notable proportion of patients. After transplantation, there was a worsening of transplant-related concerns (P < .05), depression (P < .05), and life-satisfaction (P < .001); however, pain improved (P < .01), and social functioning was well preserved. Overall, the declines in functioning after transplantation were less pronounced than anticipated. Older patients were not more compromised than younger ones; in multivariate analyses, they reported better overall quality of life (P < .01) and less depression (P < .05) before transplantation. Our findings emphasize the importance of early screening and intervention. PMID- 19135939 TI - Study of kidney function impairment after reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation. A single-center experience. AB - Acute renal failure (ARF) is a life-threatening complication after allogeneic stem cell transplantation (Allo-HSCT). Identification of ARF risk factors could be useful to develop preventive strategies for patients at high risk. The goal of this study was to evaluate the incidence and risk factors of ARF after reduced intensity conditioning Allo-HSCT (Allo-RIC). We included 188 consecutive patients who underwent Allo-RIC in our center between January 1999 and December 2006. ARF was defined as a decrease of at least 25% in baseline estimated glomerular filtration rate (GFR) calculated by modification of diet in renal disease (MDRD) equation. Conditioning consisted of fludarabine (Flu) 150 mg/m(2) in combination with busulfan (Bu) 8-10 mg/kg (n = 61), melphalan (Mel) 140 mg/m(2) (n = 115), cyclophosphamide (Cy) 120 mg/kg (n = 7) or low-dose total-body irradiation (TBI) 2 Gy (n = 5). Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine A (CsA) alone (n = 3) or in addition to methotrexate (MTX; n = 132) or mycophenolate mofetil (MMF; n = 51). The cumulative incidence of ARF at 1 year was 52% (n = 97 patients) after Allo-RIC. Most cases (86%) occurred within the first 3 months, and the main cause was the administration of CsA (71%). The risk factors associated with ARF in multivariate analysis were: administration of MTX (hazard ratio [HR] 1.9, P =.02), more than 3 lines of therapy prior to Allo-RIC (HR 1.8, P = .01), diabetes mellitus (HR 2.1, P < .01), and GVHD grade III-IV (HR 2.1, P = .015). In multivariate analysis, ARF was an independent risk factor for 1-year nonrelapse mortality (NRM) (HR 3, 95% confidence interval [CI]: 1.5-6, P = .002). Patients who experienced ARF had lower 1-year overall survival (OS; 53% versus 74%, P < .05). ARF is a frequent complication in patients after Allo-RIC, and it has a negative impact on outcome. Identification of ARF risk factors could help to avoid exposure to nephrotoxic drugs during the follow-up in patients at high risk. PMID- 19135940 TI - Allogeneic stem cell transplantation for adults with myelodysplastic syndromes: importance of pretransplant disease burden. AB - Allogeneic stem cell transplantation is the only known curative therapy for myelodysplastic syndromes (MDS). We present the transplant outcomes for 84 adult MDS patients, median age 50 (18-69 years), undergoing allogeneic hematopoietic stem cell transplantation (HSCT) at the University of Minnesota between 1995 and 2007. By WHO criteria 35 (42%) had refractory anemia with excess blasts (RAEB-1 or 2), 23 (27%) had refractory cytopenia with multilineage dysplasia (RCMD) or RCMD and ringed sideroblasts (RCMD-RS), and the remaining 26 (31%) had refractory anemia (RA), myelodysplastic syndrome-unclassifiable (MDS-U), chronic myelomonocytic leukemia (CMML), myelodysplastic/myeloproliferative disease (MDS/MPD), or myelodysplastic syndrome-not otherwise specified (MDS-NOS). Graft source was related in 47 (56%), unrelated donor (URD) marrow in 11 (13%), and unrelated cord blood (UCB) in 26 (31%). The conditioning regimen included total body irradiation (TBI) in 94% of transplantations; 52 (62%) myeloablative (MA) and 32 (38%) nonmyeloablative (NMA) regimens. Cumulative incidence of neutrophil engraftment by day +42, acute graft-versus-host disease (aGVHD) by day +100, and chronic GVHD (cGVHD) by 1 year were 88% (80%-96%, 95% confidence interval [CI]), 43% (36%-50%, 95% CI), and 15% (10%-20%, 95% CI), respectively. One-year treatment-related mortality (TRM), relapse, disease-free survival (DFS), and overall survival (OS) were 39% (28%-50%, 95% CI), 23% (12%-32%, 95% CI), 38% (28% 48%, 95% CI), and 48% (38%-58%, 95% CI) respectively. Cumulative incidence of relapse at 1 year in patients with pre-HCT complete remission (CR) or <5% blasts was improved at 18% (8%-28%, 95% CI) compared to 35% (16%-54%, 95% CI) in patients with 5%-20% blasts (P = .07). Additionally, with MA conditioning, the incidence of relapse at 1 year trended lower at 16% (6%-26%, 95% CI) versus 35% (18%-52%, 95% CI) in NMA (P = .06), and a statistically significant decrease in relapse was noted in patients entering HCT with CR or <5% blasts with an incidence of 9% (0%-18%, 95% CI) (MA) versus 31% (11%-51%, 95% CI) (NMA) (P = 0.04). For those patients with > or =5% blasts, MA conditioning did not significantly decrease relapse rates. One-year TRM was similar between MA and NMA conditioning. For patients entering transplant in CR or with <5% blasts, prior treatment to reach this level did not impact rates of relapse or transplant related mortality when all patients were analyzed; however, when broken down by conditioning intensity, there was a trend toward improved DFS in those NMA patients who were pretreated. Finally, 1-year DFS was similar using related donor peripheral blood stem cell (PBSC)/marrow, URD marrow, or UCB grafts. These data suggest that (1) blast percentage <5% at HSCT is the major predictor of improved DFS and relapse and prior treatment to reach this disease status may have value in leading to improved DFS; (2) MA conditioning is associated with lower relapse risk, particularly in patients with CR or <5% blasts, but is not able to overcome increased disease burden; (3) NMA conditioning yields equivalent TRM, DFS, and OS, and is reasonable in patients unsuited for MA conditioning; (4) the donor sources tested (PBSC, bone marrow [BM], or UCB) yielded similar outcomes. PMID- 19135941 TI - Pharmacokinetics and pharmacodynamics of plerixafor in patients with non-Hodgkin lymphoma and multiple myeloma. AB - Phase I pharmacokinetic (PK) and pharmacodynamic (PD) studies in healthy volunteers demonstrated that plerixafor (AMD3100), a CXCR4 antagonist, administered either alone or with granulocyte colony-stimulating factor (G-CSF), resulted in dose-dependent mobilization of CD34(+) cells in the peripheral blood. The purpose of this study was to evaluate the safety and the PK and PD of plerixafor with G-CSF in patients with non-Hodgkin lymphoma (NHL) and multiple myeloma (MM). This was a phase II, open-label, single-arm study conducted in 2 centers in Canada. Patients aged 18 to 70 years with NHL or MM eligible for autologous transplantation were eligible. A total of 22 patients (8 with NHL and 14 with MM) were enrolled in the study. The patients were given G-CSF (10 microg/kg/day subcutaneously [s.c.]) for 4 days in the morning and plerixafor 240 microg/kg s.c. on the evening before each day of apheresis. Apheresis was initiated 10 to 11 hours after each evening dose of plerixafor and after the morning dose of G-CSF. This regimen was repeated for up to 5 days or until > or = 5 x 10(6) CD34(+) cells/kg were collected. The objectives were to determine the safety and efficacy of plerixafor in patients with NHL and MM, and the PK and PD of a single 240-microg/kg dose of plerixafor administered after 4 days of G-CSF mobilization in these patients. The median absolute peripheral blood CD34(+) cell count increased from 24.0 cells/microL before plerixafor administration to 75.0 cells/microL before the first apheresis (10 to 11 hours after treatment with plerixafor). The median number of CD34(+) cells collected in a median of 1 day was 5.7 x 10(6) cells/kg in the patients with NHL and 12.0 x 10(6) cells/kg in those with MM. All patients underwent transplantation with prompt and durable engraftment. The PK profile of plerixafor was characterized in 13 patients (5 with NHL and 8 with MM). Overall, the PK parameters were comparable in the patients with NHL and those with MM. Plerixafor was rapidly absorbed after s.c. administration with no observable lag time, with peak plasma concentrations occurring 0.5 hour after administration in most patients. Plerixafor was rapidly cleared, with a median terminal half-life of 4.6 hours. The median maximum increase in the number of circulating cells from baseline was 4.2-fold (range, 3.0- to 5.5-fold), with the maximum fold increase occurring approximately 10 hours after plerixafor injection for all patients. The plerixafor PK and PD profiles in the study patients were consistent with those in healthy volunteers and support the current dosing regimen and timing of apheresis. Plerixafor was safe and effective in mobilizing CD34(+) cells for transplantation. PMID- 19135942 TI - Blood stream infections in allogeneic hematopoietic stem cell transplant recipients: reemergence of Gram-negative rods and increasing antibiotic resistance. AB - Blood stream infections (BSI) are a well-known cause of morbidity and mortality in hematopoietic stem cell transplant (HSCT) patients. The aim of this study was to analyze etiology and microbial resistance of BSI in patients undergoing allogeneic HSCT in a single center over a 4-year period (2004-2007). There were 168 episodes of BSI in 132 patients (median 10 days after HSCT) and 182 pathogens were isolated. Gram-positive bacteria (GPB) accounted for 57% of 182 isolates. Gram-negative rods (GNR) for 37% and fungi for 6%. All patients received routine fluoroquinolone prophylaxis. There was a significant decrease in GPB/GNR ratio over time, from 2.4 in 2004 to 1 in 2007 (P = .043). Among GPB, staphylococci decreased from 37 of 68 (64%) in 2004-2005 to 8 of 35 (23%) in 2006-2007 (P < .002). The Enterococcus faecalis/E. faecium ratio decreased from 4.5 in 2004 to 0.33 in 2007 (P = .006), whereas the total number of enterococcal strains per year did not change. The incidence of Escherichia coli among GNR increased from 3 of 15 (20%) in 2004 to 13 of 21 (62%) in 2007 (P = .003). Fluoroquinolone resistance was common, both among GPB and GNR (81% and 74%, respectively). Mortality rate at 7 days after BSI was 11% (19 of 168), reaching 39% for Pseudomonas aeruginosa BSI (7 of 18). BSI remains a frequent and potentially life threatening complication of allogeneic HSCT, the causative organism influencing 7 and 30-day mortality rate. BSI etiology may change rapidly, requiring implementation of new empirical-therapy schemes. PMID- 19135943 TI - Donor and recipient CMV serostatus and outcome of pediatric allogeneic HSCT for acute leukemia in the era of CMV-preemptive therapy. AB - In the era of cytomegalovirus (CMV)-preemptive therapy, it is unclear whether CMV serostatus of donor or recipient affects outcome of allogeneic hematopoietic stem cell transplantation (HSCT) among children with leukemia. To investigate, consecutive patients aged 0-18 who underwent primary HSCT for acute leukemia in 1997-2007 (HLA-matched sibling or unrelated donor, myeloablative conditioning, unmanipulated bone marrow or peripheral blood, preemptive therapy, no CMV prophylaxis) were followed retrospectively through January 2008. Treatment failure (relapse or death) was analyzed using survival-based proportional hazards regression. Competing risks (relapse and nonrelapse mortality, NRM) were analyzed using generalized linear models of cumulative incidence-based proportional hazards. Excluding 4 (2.8%) patients lacking serostatus of donor or recipient, there were 140 subjects, of whom 50 relapsed and 24 died in remission. Pretransplant CMV seroprevalence was 55.7% in recipients, 57.1% in donors. Thirty five (25.0%) grafts were from seronegative donor to seronegative recipient (D-/R ). On univariate analysis, D-/R- grafts were associated with shorter relapse-free survival (RFS) than other grafts (median 1.06 versus 3.15 years, P < .05). Adjusted for donor type, diagnosis, disease stage, recipient and donor age, female-to-male graft, graft source, and year, D-/R- graft was associated with relapse (hazards ratio 3.15, 95% confidence interval 1.46-6.76) and treatment failure (2.45, 1.46-4.12) but not significantly with NRM (2.00, 0.44-9.09). In the current era, children who undergo allogeneic HSCT for acute leukemia have reduced risk of relapse and superior RFS when recipient and/or donor is CMV seropositive before transplantation. However, no net improvement in RFS would be gained from substituting seropositive unrelated for seronegative sibling donors. PMID- 19135944 TI - Hematopoietic cell transplantation from an HLA-mismatched familial donor is feasible without ex vivo-T cell depletion after reduced-intensity conditioning with busulfan, fludarabine, and antithymocyte globulin. AB - To extend the use of allogeneic hematopoietic cell transplantation (HCT) to patients without an HLA-matched donor, we investigated HCT from a related donor with 1 fully mismatched HLA-haplotype after conditioning with busulfan in reduced dose, fludarabine, and antithymocyte globulin. Hematopoietic cells were collected from the donors via leukapheresis after mobilization and infused without further manipulation. Cyclosporin and methotrexate were administered for graft-versus host disease (GVHD) prophylaxis. Posttransplant engraftment, GVHD, and transplantation-related mortality (TRM) were recorded. Thirty-one patients (age range: 16-69 years) with high-risk acute leukemia/myelodysplastic syndrome (n = 25) or bone marrow failure (n = 6) were enrolled. The donors were either mothers (n = 14), offspring (n = 9), or siblings (n = 8) of these patients. Excluding 3 patients who died or relapsed with leukemia within 3 weeks after HCT, all the remaining 28 patients engrafted with neutrophils (>500/microL) at a median of 16.5 days. Twenty-two of 24 evaluated patients achieved complete donor chimerism (> or =95%) 2 weeks after HCT and none experienced graft failure subsequently. The cumulative incidences of grade 2-4 acute GVHD (aGVHD) and moderate-severe chronic GVHD (cGVHD) were 19% (95% confidence interval [CI], 9%-40%) and 20% (95% CI, 10%-41%), respectively. After a median follow-up of 18.2 months (range: 6.3 52.1), 18 patients remained alive (53%). Four patients died without recurrence/progression of underlying diseases giving a TRM of 13% (95% CI, 5% 33%). HCT from an HLA-mismatched family member is feasible without ex vivo T cell depletion when reduced-intensity conditioning containing anti-hymocyte globulin is performed. PMID- 19135945 TI - Possible association between obesity and posttransplantation complications including infectious diseases and acute graft-versus-host disease. AB - Both obesity and malnutrition are considered risk factors for complications after bone marrow transplantation (BMT). To elucidate the impact of pretransplantation body mass index (BMI) on clinical outcome, we performed a retrospective cohort study with registration data from the Japan Marrow Donor Program (JMDP). Between January 1998 and December 2005, a total of 3935 patients received unrelated BMT through the JMDP; of these, 3827 patients for whom pretransplantation height and weight data were available were included in the study. Patients were stratified according to pretransplantation BMI values (low BMI: BMI < 18 kg/m(2), n = 295; normal BMI: 18 < or = BMI < 25 kg/m(2), n = 2906; overweight: 25 < or = BMI <30 kg/m(2), n = 565; obese: 30 kg/m(2) < or = BMI, n = 61). In a univariate analysis, pretransplantation BMI was associated with a significantly greater risk of grade II-IV acute graft-versus-host disease (GVHD; P = .03). Multivariate analysis showed that pretransplantation BMI tended to be associated with an increased risk of grade II-IV acute GVHD (P = .07). Obesity was associated with an increased risk of infection compared with normal BMI (odds ratio = 1.9; 95% confidence interval = 1.1 to 3.2; P = .02). Our findings demonstrate a correlation between pretransplantation BMI and posttransplantation complications. Although BMI depends strongly on multiple factors, the effect of obesity on clinical outcome should be evaluated in a prospective study. PMID- 19135947 TI - Early central nervous complications after umbilical cord blood transplantation for adults. AB - Early central nervous complications (CNS) are significant after allogeneic stem cell transplantation; however, the clinical characteristics of early CNS complications have not yet been well described. The medical record of 77 patients who underwent cord blood transplantation (CBT) between March 2001 and November 2005, at 8 centers of the Nagoya Blood and Marrow Transplantation Group were retrospectively reviewed. The preparative regimen included myeloablative CBT (n = 31) or reduced-intensity (RI)-CBT (n = 46). Of the 77 patients, 10 (13%) developed early CNS complications. Causes included Cyclosporine encephalopathy (n = 5), tacrolimus encephalopathy (n = 2), thrombocytic microangiopathy (n = 1), and unknown (n = 3). The median time of onset was 19 days (range: 2-58 days). All of the 10 patients developed impaired consciousness. Seizures developed in 6 patients. Early CNS complications spontaneously subsided in 3 patients. Three patients responded to cyclosporine or tacrolimus discontinuation. The remaining 4 patients died within 30 days of developing of early CNS complications. No relationship was detected between the preparative regimen and the onset of early CNS complications, while an HLA disparity showed borderline significance (hazard ratio, 3.24; 95% confidential interval, 0.94-11.20; P = .06). Early CNS complications are a significant problem after CBT, and the clinician has to be aware of the possibility of these complications. PMID- 19135946 TI - A Phase II trial of autologous stem cell transplantation followed by mini allogeneic stem cell transplantation for the treatment of multiple myeloma: an analysis of Eastern Cooperative Oncology Group ECOG E4A98 and E1A97. AB - Conventional allogeneic hematopoietic stem cell transplantation (HSCT) for multiple myeloma is associated with high transplantation-related mortality (TRM). Nonmyeloablative allogeneic transplantation (NST) uses the well-known graft versus-myeloma (GVM) effect to eradicate minimal residual disease. The Eastern Cooperative Oncology Group conducted a Phase II trial of autologous HSCT followed by NST to provide maximal tumor cytoreduction to allow for a subsequent GVM effect. Patients received melphalan 200 mg/m(2) with autologous HSCT, followed by fludarabine 30 mg/m(2) in 5 daily doses and cyclophosphamide 1 g/m(2) in 2 daily doses with matched sibling donor NST. Graft-versus-host disease (GVHD) prophylaxis included cyclosporine and corticosteroids. The primary endpoints were TRM, graft failure, acute GVHD, progression-free survival (PFS), and overall survival (OS). Thirty-two patients were enrolled into the study; 23 patients completed both transplantations (72%). Best responses post-NST were 7 (30%) complete remission (CR), 11 (48%) partial remission (PR), 2 (9%) no response, and 3 (13%) not evaluable. Acute grade III-IV GVHD was observed in 4 patients (17%), and chronic GVHD was seen in 13 patients (57%; 7 limited, 6 extensive). Chronic GVHD resulted in the following responses: 3 (23%) CR, 1 continuing CR, and 6 (46%) PR. Two patients (8.7%) had early TRM. With a median follow up of 4.6 years, the median PFS was 3.6 years, and the 2-year OS was 78%. Our findings indicate that autologous HSCT followed by NST is feasible, with a low early TRM in a cooperative group setting. The overall response rate was 78%, including 30% CR, similar to other reports for autologous HSCT-NST. Because a plateau in PFS or OS was not observed with this treatment approach even in patients achieving CR, we suggest that future studies use posttransplantation maintenance therapy. PMID- 19135948 TI - Graft-versus-host disease prophylaxis with everolimus and tacrolimus is associated with a high incidence of sinusoidal obstruction syndrome and microangiopathy: results of the EVTAC trial. AB - A calcineurin inhibitor combined with methotrexate is the standard prophylaxis for graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). Everolimus, a derivative of sirolimus, seems to mediate antileukemia effects. We report on a combination of everolimus and tacrolimus in 24 patients (median age, 62 years) with either myelodysplastic syndrome (MDS; n = 17) or acute myeloid leukemia (AML; n = 7) undergoing intensive conditioning followed by HSCT from related (n = 4) or unrelated (n = 20) donors. All patients engrafted, and only 1 patient experienced grade IV mucositis. Nine patients (37%) developed acute grade II-IV GVHD, and 11 of 17 evaluable patients (64%) developed chronic extensive GVHD. Transplantation-associated microangiopathy (TMA) occurred in 7 patients (29%), with 2 cases of acute renal failure. The study was terminated prematurely because an additional 6 patients (25%) developed sinusoidal obstruction syndrome (SOS), which was fatal in 2 cases. With a median follow-up of 26 months, the 2-year overall survival rate was 47%. Although this new combination appears to be effective as a prophylactic regimen for acute GVHD, the incidence of TMA and SOS is considerably higher than seen with other regimens. PMID- 19135949 TI - Unrelated donor reduced-intensity allogeneic hematopoietic stem cell transplantation for relapsed and refractory Hodgkin lymphoma. AB - Myeloablative allogeneic hematopoietic cell transplantation (HCT) may cure patients with relapsed or refractory Hodgkin lymphoma (HL), but is associated with a high treatment-related mortality (TRM). Reduced-intensity and nonmyeloablative (RIC/NST) conditioning regimens aim to lower TRM. We analyzed the outcomes of 143 patients undergoing unrelated donor RIC/NST HCT for relapsed and refractory HL between 1999 and 2004 reported to the Center for International Blood and Marrow Transplant Research (CIBMTR). Patients were heavily pretreated, including autologous HCT in 89%. With a median follow-up of 25 months, the probability of TRM at day 100 and 2 years was 15% (95% confidence interval [CI] 10%-21%) and 33% (95% CI 25%-41%), respectively. The probabilities of progression free survival (PFS) and overall survival (OS) were 30% and 56% at 1 year and 20% and 37% at 2 years. The presence of extranodal disease and the Karnofsky Performance Scale (KPS) <90 were significant risk factors for TRM, PFS, and OS, whereas chemosensitivity at transplantation was not. Dose intensity of the conditioning regimen (RIC versus NST) did not impact outcomes. Unrelated donor HCT with RIC/NST can salvage some patients with relapsed/refractory HL, but relapse remains a common reason for treatment failure. Clinical studies should be aimed at reducing the incidence of acute graft-versus-host disease (GVHD) and relapse. PMID- 19135950 TI - Prognostic factors and clinical outcomes of high-dose chemotherapy followed by autologous stem cell transplantation in patients with peripheral T cell lymphoma, unspecified: complete remission at transplantation and the prognostic index of peripheral T cell lymphoma are the major factors predictive of outcome. AB - High-dose chemotherapy followed by autologous stem cell transplantation (HDT/ASCT) offers a rescue option for T cell lymphoma patients with poor prognosis. However, the effectiveness of HDT/ASCT in patients with various peripheral T cell subtypes, optimal transplant timing, and the prognostic factors that predict better outcomes, have not been identified. We retrospectively investigated the clinical outcomes and prognostic factors for HDT/ASCT in 64 Korean patients with peripheral T cell lymphoma, unspecified (PTCL-U) between March 1995 and February 2007. The median age at transplantation was 44 years (range: 15-63 years). According to the age-adjusted International Prognostic Index (a-IPI) and the prognostic index of PTCL (PIT), 8 patients (12.5%) were in the high-risk group and 16 (26.6%) had the 2-3 PIT factors, respectively. After a median follow-up of 29.7 months, the 3-year overall survival (OS) and progression free survival (PFS) rates were 53.0% +/- 7.5% and 44.3% +/- 7.0%, respectively. Univariate analysis showed that poor performance status, high lactate dehydrogenase (LDH) levels, high a-IPI score, high PIT classes, failure to achieve complete response (CR) at transplantation, and nonfrontline transplantation were associated with poor OS. Multivariate analysis showed that failure to achieve CR at transplantation (hazard ratio [HR] 2.23; 95% confidence interval [CI] 1.78-7.93) and 2-3 PIT factors (HR 3.76; 95% CI 1.02-5.42) were independent prognostic factors for OS. Failure to achieve CR at transplantation and high PIT are negative predictable factors for survival following HDT/ASCT in patients with PTCL-U. PMID- 19135951 TI - Donor treatment with a multipegylated G-CSF maximizes graft-versus-leukemia effects. AB - Donor treatment with granulocyle-colony stimulating factor (G-CSF) is known to modulate immune function, characterized by the generation of regulatory myelogenous and T cell populations and Th2 differentiation. Recently, these effects have been shown to be enhanced by pegylation of the G-CSF molecule, which also improves graft-versus-leukemia (GVL) via activation of invariant natural killer (iNK) T cells. We have compared G-CSF bound to a single PEG molecule (monopeg-G-CSF) as used clinically to a G-CSF molecule bound to multiple PEG molecules (multipeg-G-CSF) in major histocompatibility complex (MHC) disparate and matched models of graft-versus-host disease (GVHD) and GVL. We demonstrate that multipeg-G-CSF induces greater levels of progenitor cell, myelogenous, and iNKT cell expansion than monopeg-G-CSF, while inducing similar protection from GVHD. Despite this, multipeg-G-CSF enhanced CTL function in vivo and improved iNKT cell-dependent leukemia clearance. Thus, GVL and GVHD can be further separated after allogeneic stem cell transplantation by mobilization with a multiple-pegylated G-CSF molecule. PMID- 19135952 TI - Improving homeopathic prescribing. PMID- 19135953 TI - Cost-benefit evaluation of homeopathic versus conventional therapy in respiratory diseases. AB - METHODS: A retrospective observational study was conducted on 105 out of 233 patients suffering from chronic respiratory disease attending the Homeopathic Clinic of the Campo di Marte Hospital in Lucca (Tuscany, Italy) between October 1998 and May 2003. We assessed the cost of conventional medicinal products using Anatomic Therapeutic Chemical (ATC) classification, specific for the pathology in question, and the general costs in the year preceding the first appointment at the Homeopathic Clinic vs. the first and second year subsequent to homeopathic treatment. The costs of conventional drugs for a group of patients affected by asthma (8 patients) and recurrent respiratory infections (16 patients) with long term use of conventional medicine treated by homeopathy were compared with the expenses of conventional drugs of a matched group of 16 and 32 patients, respectively. RESULTS: Costs of pharmacological therapy specific for respiratory diseases were reduced by 46.3% (n=105) in the first year (P<0.01); and by 47.5% (n=72) in the second year (P<0.01) of homeopathic treatment. Reduction in general drug costs during homeopathic therapy was 42.4% in the first year (P<0.01); and 49.8 in the second year (N.S.). Costs for patients affected by chronic asthma showed a reduction in expenses of 71.1% for specific medicines relative to the group in homeopathic treatment vs. an increase of 12.3% in the group treated only with conventional drugs after the first year of follow-up and, respectively, a reduction of -54.4% for homeopathic treatment vs. +45.2% after the second year. For patients with recurrent respiratory infections we found a reduction of 35.8% in the homeopathic group in the first year, compared to an increase 8.6% of costs for specific drugs in the control group; in the second year the respective figures were -43.6% versus +7.8% in the control group. CONCLUSIONS: Homeopathic treatment for respiratory diseases (asthma, allergic complaints, Acute Recurrent Respiratory Infections) was associated with a significant reduction in the use and costs of conventional drugs. Costs for homeopathic therapy are significantly lower than those for conventional pharmacological therapy. PMID- 19135954 TI - Evaluation of the quality of life after individualized homeopathic treatment for seasonal allergic rhinitis. A prospective, open, non-comparative study. AB - BACKGROUND: Quality of life (QoL) is an important outcome measure in the treatment of seasonal allergic rhinitis (SAR), a condition for which homeopathy is frequently used. OBJECTIVE: The assessment of the effect of homeopathic medical prescriptions with the Rhino-conjunctivitis Quality of Life Questionnaire (RQLQ) in the treatment of SAR. METHODS: A prospective, open, non-comparative study was conducted in Belgium. Patients aged between 14 and 68 years with SAR were treated by one of seven homeopathic physicians. Patients completed the RQLQ at baseline and again after three and four weeks of homeopathic treatment. RESULTS: Seventy-four patients were screened, of whom 46 met the study eligibility criteria (average age 36 years, 70% female). The mean RQLQ score at baseline was 3.40 (+/-.98). After three and four weeks of homeopathic treatment it had fallen to 1.97 (+/-1.32) (P=0.0001), and 1.6 (+/-1.28) (P=0.0001), respectively. CONCLUSIONS: After homeopathic treatment, patients reported an alleviation of their symptoms of allergic rhinitis as reported in the RQLQ. A formal Randomized Clinical Trial (RCT) is indicated. PMID- 19135955 TI - Observational study of homeopathic and conventional therapies in patients with diabetic polyneuropathy. AB - METHODS: The feasibility and outcomes of homeopathic therapy in a group of type-2 diabetes mellitus patients with diabetic neuropathy were studied in a prospective observational study. Patients were followed from baseline (T0) for 6 months (T1) and for 12 months (T2), treatment was adjusted as necessary. Primary outcome was diabetic neuropathy symptom (DNS) score, secondary outcomes were clinical evolution and short-form-36 (SF-36)-evaluated quality of life (QOL). RESULTS: Homeopathy was used in 45 patients, 32 of whom completed the observation study, and in parallel the conventional therapy outcomes were observed in 32 patients, 29 of whom completed the study. DNS improved in both groups during the observation period, but the change with respect to baseline was statistically significant only in Homeopathic group at T1 (P=0.016). Over the course of the observation there was a substantial stability of the electroneurophysiological values, blood pressure and body weight in both groups, a slight decrease of fasting blood glucose and glycated haemoglobin in Homeopathic group. QOL scores showed an improvement in Homeopathic group only. The cost of conventional drugs decreased in Homeopathic group from 114 euro/month to 94 euro/month at T1. CONCLUSION: Complementary homeopathic therapy of diabetic neuropathy was feasible and promising effects in symptom scores and cost savings were observed. PMID- 19135956 TI - Statistical analysis of six repertory rubrics after prospective assessment applying Bayes' theorem. AB - BACKGROUND: After prospective assessment of six homeopathic symptoms we validated some rubrics of the homeopathic repertory using Bayesian theory. In this paper we introduce statistical arguments for introducing or discarding entries from the repertory. METHODS: 4094 patients entered the prospective study and 4072 prescriptions were evaluated. After translating typeface into Likelihood Ratios (LRs), Confidence Intervals and the probability of existing repertory entries compared to our findings were calculated. OUTCOME: Our assessment yielded 121 relevant results to validate existing repertory entries. Five symptoms could be compared with Kent's original repertory; they have about the same prevalence (range 3.9-6.5%) in the whole population, but the size of the corresponding repertory rubrics varies from 3 to 103 entries. LR assessment reduced the larger rubrics and supplemented the smaller ones. Our results do not correspond with 56% of the existing repertory entries regarding five symptom-rubrics. This result cannot be generalised for the whole repertory. PMID- 19135957 TI - Homeopathic drug selection using Intuitionistic fuzzy sets. AB - Using intuitionistic fuzzy set theory, Sanchez's approach to medical diagnosis has been applied to the problem of selection of single remedy from homeopathic repertorization. Two types of Intuitionistic Fuzzy Relations (IFRs) and three types of selection indices are discussed. I also propose a new repertory exploiting the benefits of soft-intelligence. PMID- 19135958 TI - Ignatia in the treatment of oral lichen planus. AB - OBJECTIVE: To evaluate the effectiveness of Ignatia homeopathic 30C in management of oral lichen planus (OLP). METHODS AND MATERIALS: In this single blind randomized control clinical trial, 30 consecutive patients with oral lesions consistent clinically and histologically with erosive and/or atrophic OLP were recruited. The patients were randomly divided into two groups to receive Ignatia or placebo. They were treated for 4 months. RESULTS: Mean lesion sizes and mean pain measures differed between control and treatment groups favouring Ignatia (p<0.05). CONCLUSION: Our results suggest that Ignatia has a beneficial effect in treatment of OLP in selected patients. PMID- 19135959 TI - Lymphocyte proliferation stimulated by activated human macrophages treated with Canova. AB - INTRODUCTION: Canova (CA) is a homeopathic medication with immunomodulatory properties, recommended for patients with a depressed immune system. CA has been reported to increase in leukocyte numbers, cellular differentiation and reduction in tumor size. AIM AND METHOD: Since CA may stimulate lymphocyte differentiation, proliferation, and/or survival, the aim of the present study was to compare the mitotic index (MI) of phytohemagglutinin-stimulated human lymphocytes cultured in a medium supplemented with human macrophages activated by CA, with lymphocytes cultured in a medium without CA-treated macrophages. RESULTS: In this study, the MI of lymphocyte cultured received the medium containing CA-stimulated macrophages showed a higher proliferation index (p<0.01) than the lymphocytes cultured in a medium without CA-treated macrophages. Our results suggest that CA treatment, in addition to activating macrophages, indirectly induces lymphocyte proliferation and has potential as a new adjuvant therapeutic approach. PMID- 19135960 TI - Polarity analysis, a new approach to increase the precision of homeopathic prescriptions. AB - INTRODUCTION: The Swiss randomised controlled trial of homeopathy for attention deficit hyperactivity disorder (ADHD) was a rigorous test of homeopathy. In each of its three phases it delivered evidence for a specific effect of homeopathic treatment, but it also unmasked weaknesses of the method. Misleading reports of sensations and mind symptoms by parents were frequent, while modalities and polar symptoms usually proved to be reliable information for repertorisation. The problem of cases with a paucity of symptoms was resolved by reintroduction of (pathognomonic) perception symptoms into the repertorisation. Additionally polarity analysis, a further development of Boenninghausen's concept of contraindications, was tested and introduced. It allows a precise differential diagnosis of possible homeopathic medicines. Increasing the rate of optimal prescriptions by 20%, polarity analysis turned out to be the most efficient modification to case analysis. This paper describes the transfer of the new insights to the treatment of other diseases and as the evaluation of this process. METHODS: Polarity analysis was tested and applied in acute diseases by completing patient histories with repertory specific checklists, mainly based on modalities and polar symptoms. The checklists encompassed eleven different complaints. Treatment results were compared with results reached by conventional homeopathic case analysis methods. The same procedure was applied in chronic diseases with repertory-specific questionnaires. Again, eleven different areas were covered. Treatment results for chronic diseases were also compared with a conventional case analysis approach. RESULTS: Polarity analysis, checklists and questionnaires led to an increase in optimal prescriptions of 22% in acute diseases and 16% in chronic diseases. In addition, the average improvement rates in chronic disease were 9% higher than with conventional homeopathic procedures. The new method is demonstrated by a case example with a verified clinical cure, and its impacts on homeopathy are discussed. CONCLUSION: The use of polarity analysis as an integral part of case analysis and differential diagnosis of possible remedies together with an increased awareness for assessing the reliability of symptoms in repertorisation lead to a substantial improvement in the precision of homeopathic prescriptions. PMID- 19135961 TI - Treatment of spasmodic dysphonia with homeopathic medicine: a clinical case report. AB - Botulinum toxin (Botox) injection is the only conventional medical treatment available for patients with spasmodic dysphonia (SD). Some patients are reluctant to receive Botox treatment due to concerns about unknown long-term side effects, expense, and dependence on repeated injections. The purpose of the study was to report the perceptual and physiological changes in the vocal functions of an SD patient treated with classical homeopathy. The results were similar to a previous case report: classical homeopathy seems to be capable of amelioratiny SD symptoms beyond the short-term effects of Botox injections. Although the physiological mechanism of homeopathic healing is not fully accounted for by the current bio medical models, it may be an effective therapeutic alternative for some SD patients. PMID- 19135962 TI - In search of the reliable repertory. AB - The development of homeopathic repertories is complex, reflecting history, the emergence of divergent views on homeopathic philosophy, and differences in opinion as to what constitutes reliable materia medica. The purpose of this paper is to critically evaluate the content of repertories examining its reliability, the quality of source material, and the evidence that it forms a reliable bridge between case and materia medica. Reliability may be improved by demanding higher standards and consistency of evidence. However, it is necessary to understand what constitutes evidence, and the importance of taking into account the context in which practitioners use the repertory. This paper will suggest that rather than demanding certain 'standards', practitioners will be better served by a greater understanding of the sources of knowledge and by reflexivity of the key players in the construction of our repertories. 'The repertory' is considered generally here as the deconstruction of different repertories. The strengths and weaknesses in particular, whilst interesting, would be the topic of another a paper in its own right. Where individual repertories are mentioned, they are referred to as examples only. PMID- 19135963 TI - 20 years ago: The British Homoeopathic Journal, January 1989. PMID- 19135965 TI - Homeopathy: the clinical picture is no clearer. PMID- 19135966 TI - Are media attacks justified? A response to Vithoulkas. PMID- 19135970 TI - Ionic liquids promote amyloid formation from alpha-synuclein. AB - The slow process required for alpha-synuclein to form amyloid fibrils is a major obstacle in the development of therapeutic compounds for alpha-synuclein-related neurodegenerative diseases such as Parkinson's disease (PD). Here we have developed an efficient method by which amyloid fibrils can be formed from alpha synuclein using ionic liquids (ILs). This report indicates that ILs could potentially be used as a stimulator for the amyloid formation of alpha-synuclein. PMID- 19135969 TI - Visual, base-specific detection of nucleic acid hybridization using polymerization-based amplification. AB - Polymerization-based signal amplification offers sensitive visualization of biotinylated biomolecules functionalized to glass microarrays in a manner suitable for point-of-care use. Here we report using this method for visual detection of multiplexed nucleic acid hybridizations from complex media and develop an application toward point mutation detection and single nucleotide polymorphism (SNP) typing. Primer extension reactions were employed to label selectively and universally all complementary surface DNA hybrids with photoinitiators, permitting simultaneous and dynamic photopolymerization from positive sites to 0.5-nM target concentrations. Dramatic improvements in signal ratios between complementary and mismatched hybrids enabled visual discrimination of single base differences in KRAS codon-12 biomarkers. PMID- 19135971 TI - Use of tandem Biacore-mass spectrometry to identify platelet membrane targets of novel monoclonal antibodies. AB - The monoclonal antibodies (mAbs) ALMA.17 and ALMA.7 recognize human platelet membrane proteins. ALMA.17 is directed against alpha(IIb)beta(3) integrin, but the target of ALMA.7 was unknown previously. Tandem Biacore micropurification and mass spectrometry (MS) analysis of a platelet membrane lysate was used to identify the target of ALMA.7. Detergent lysates enriched in membrane proteins were perfused over immobilized ALMA.17 or ALMA.7 in a Biacore system. The captured proteins were eluted, concentrated on C3 magnetic beads, and digested with trypsin before nano liquid chromatography-tandem mass spectrometry (LC MS/MS) analysis. Critical adjustments needed to be made in (i) the detergent mixture to preserve protein antigenicity and sensor chip integrity and (ii) the method of trypsin digestion to concentrate the proteins and use elution buffers that do not interfere with MS. The target of ALMA.17 was confirmed to be alpha(IIb)beta(3) integrin, whereas that of ALMA.7 was identified as CD226 (PTA 1, DNAM-1, TLiSa-1). This was confirmed by immunoassays comparing ALMA.7 with a commercial anti-CD226 mAb. Thus, a tandem Biacore and nano LC-MS/MS strategy allowed unambiguous identification of an unknown antigen in a complex medium such as a platelet membrane lysate. This strategy may be employed to identify any protein "capturable" on a sensor chip provided that one uses appropriate experimental conditions. PMID- 19135972 TI - Age-related differences in oxidative protein-damage in young and senescent fibroblasts. AB - Aging is accompanied by an accumulation of oxidized proteins and cross-linked modified protein material. The intracellular formation and accumulation of highly oxidized and cross-linked proteins, the so-called lipofuscin, is a typical sign of senescence. However, little is known whether the lipofuscin accumulation during aging is related to environmental conditions, as oxidative stress, and whether the accumulation of oxidized proteins and lipofuscin is preferentially taking place in the cytosol or the nucleus and finally, what is the role of lysosomes in this process. Therefore, we investigated human skin fibroblasts in an early stage of proliferation ("young cells") and in a late stage ("senescent cells"). Such cells were compared for the amount of protein carbonyls and lipofuscin and their distribution within the cytosol and the nucleus. Furthermore, cells were exposed to single and repeated doses of hydrogen peroxide and paraquat, measuring the same set of parameters. In addition to that the role of the proteasome to degrade oxidized proteins in young and senescent cells was tested. Furthermore, detailed microscopic analysis was performed testing the intracellular distribution of lipofuscin. The results clearly demonstrated that repeated/chronic oxidative stress induces a senescence-like phenotype of the distribution of oxidized proteins as well as of lipofuscin. It could be demonstrated that most of the lipofuscin is located in lysosomes and that senescent cells contain less lysosomes not lipofuscin-laden in comparison to young cells. PMID- 19135973 TI - Antifolate/folate-activated HGF/c-Met signalling pathways in mouse kidneys-the putative role of their downstream effectors in cross-talk with androgen receptor. AB - This in vivo study of mouse kidneys was focused on the identification of protein mediators involved in the cross-talk between two signalling pathways. One pathway was triggered by testosterone via an androgen receptor, AR, and the other induced by CB 3717/folate via HGF, and its membrane receptor c-Met. Sequential activation of these pathways leads to a drastic decrease of testosterone-induced ornithine decarboxylase, ODC, expression. We proved that CB 3717/folate-induced ODC expression is Akt-dependent. CB 3717/folate activates Akt and ERK1/2 kinases, PTEN phosphatase and also up-regulates cyclin D2 and PCNA, but decreases GSK3beta and cyclin D1 protein levels. Testosterone activation of AR induces GSK3beta and PTEN. Results of the sequential activation of the studied signalling pathways suggest that Akt, GSK3beta and possibly ERK1/2 kinases may participate in the negative cross-talk and attenuation of AR transactivity, while the involvement of PTEN and cyclin D1 seems to be doubtful. PMID- 19135975 TI - Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase. AB - In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier. PMID- 19135974 TI - Predicting protein folding cores by empirical potential functions. AB - Theoretical and in vitro experiments suggest that protein folding cores form early in the process of folding, and that proteins may have evolved to optimize both folding speed and native-state stability. In our previous work (Chen et al., Structure, 14 (2006) 1401), we developed a set of empirical potential functions and used them to analyze interaction energies among secondary-structure elements in two beta-sandwich proteins. Our work on this group of proteins demonstrated that the predicted folding core also harbors residues that form native-like interactions early in the folding reaction. In the current work, we have tested our empirical potential functions on structurally-different proteins for which the folding cores have been revealed by protein hydrogen-deuterium exchange experiments. Using a set of 29 unrelated proteins, which have been extensively studied in the literature, we demonstrate that the average prediction result from our method is significantly better than predictions based on other computational methods. Our study is an important step towards the ultimate goal of understanding the correlation between folding cores and native structures. PMID- 19135976 TI - Molecular and functional characterization of choline transporter in human colon carcinoma HT-29 cells. AB - We examined the molecular and functional characterization of choline uptake in human colon carcinomas using the cell line HT-29. Furthermore, we explored the possible correlation between choline uptake and cell proliferation. Choline uptake was saturable and mediated by a single transport system. Interestingly, removal of Na(+) from the uptake buffer strongly enhanced choline uptake. This increase in component of choline uptake under Na(+)-free conditions was inhibited by a Na(+)/H(+) exchanger 1 (NHE1) inhibitor. Collapse of the plasma-membrane H(+) electrochemical gradient by a protonophore inhibited choline uptake. Choline uptake was inhibited by the choline analogue hemicholinium-3 (HC-3) and various organic cations, and was significantly decreased by acidification of the extracellular medium and by intracellular alkalinization. Real-time PCR revealed that choline transporter-like protein 1 (CTL1), CTL2, CTL4 and NHE1 mRNA are mainly expressed in HT-29 cells. Western blot and immunocytochemical analysis indicated that CTL1 protein was expressed in plasma membrane. The biochemical and pharmacological data indicated that CTL1 is functionally expressed in HT-29 cells and is responsible for choline uptake in these cells. We conclude that choline transporters, especially CTL1, use a directed H(+) gradient as a driving force, and its transport functions in co-operation with NHE1. Finally, cell proliferation was inhibited by HC-3 and tetrahexylammonium chloride (THA), which strongly inhibits choline uptake. Identification of this novel CTL1-mediated choline uptake system provides a potential new target for therapeutic intervention. PMID- 19135977 TI - Inhibition of vascular endothelial growth factor expression by TRUE gene silencing. AB - Pathogenic angiogenesis in various diseases including cancer, autoimmune diseases, and age-related macular degeneration is thought to be regressed with anti-angiogenic drugs. TRUE gene silencing is a new technology to eliminate a specific mRNA using synthetic sgRNA and cellular tRNase Z(L). To discover anti angiogenic sgRNAs, we applied TRUE silencing to the VEGF gene. We examined eight sgRNAs for efficacy in targeting exogenous human VEGF mRNA. Many of them worked efficiently in 293 and HeLa cells. Two of them downregulated the endogenous VEGF gene expression in HeLa cells very efficiently, and the efficacy of these two sgRNAs surpassed that of siRNA extremely. PMID- 19135978 TI - IFN-gamma down-regulates Secretoglobin 3A1 gene expression. AB - STAT1 mediates Interferon (IFN)-dependent positive and negative regulation of inflammatory gene expression in lung. In this study, we examined the effect of IFN-gamma on the expression of SCGB3A1 which is thought to play crucial roles in inflammation and epithelial cell differentiation in lung. We found that expression of SCGB3A1 was down-regulated by IFN-gamma in a time- and dose dependent manner in the murine transformed Clara Cells (mtCC) line. IFN-gamma induced the phosphorylation of STAT1, which binds to a STAT-binding element (SBE) in the SCGB3A1 gene promoter, leading to decreased transcriptional activation of this gene. PMID- 19135979 TI - TRPA1 activation by lidocaine in nerve terminals results in glutamate release increase. AB - We examined the effects of local anesthetics lidocaine and procaine on glutamatergic spontaneous excitatory transmission in substantia gelatinosa (SG) neurons in adult rat spinal cord slices with whole-cell patch-clamp techniques. Bath-applied lidocaine (1-5 mM) dose-dependently and reversibly increased the frequency but not the amplitude of spontaneous excitatory postsynaptic current (sEPSC) in SG neurons. Lidocaine activity was unaffected by the Na(+)-channel blocker, tetrodotoxin, and the TRPV1 antagonist, capsazepine, but was inhibited by the TRP antagonist, ruthenium red. In the same neuron, the TRPA1 agonist, allyl isothiocyanate, and lidocaine both increased sEPSC frequency. In contrast, procaine did not produce presynaptic enhancement. These results indicate that lidocaine activates TRPA1 in nerve terminals presynaptic to SG neurons to increase the spontaneous release of L-glutamate. PMID- 19135980 TI - MicroRNA-15b regulates cell cycle progression by targeting cyclins in glioma cells. AB - MicroRNAs (miRNAs) are non-protein-coding RNAs that function as post transcriptional gene regulators. Recent evidence has shown that miRNA plays a pivotal role in the development of many cancers including glioma, a lethal brain cancer. We have recently compared the miRNA expression profiles between normal brain and glioma tissues from Chinese patients by miRNA microarray and identified a panel of differentially expressed miRNAs. Here, we studied the function of one miRNA, miR-15b, in glioma carcinogenesis and elucidated its downstream targets. Over-expression of miR-15b resulted in cell cycle arrest at G0/G1 phase while suppression of miR-15b expression resulted in a decrease of cell populations in G0/G1 and a corresponding increase of cell populations in S phase. We further showed that CCNE1 (encoding cyclin E1) is one of the downstream targets of miR 15b. Taken together, our findings indicate that miR-15b regulates cell cycle progression in glioma cells by targeting cell cycle-related molecules. PMID- 19135981 TI - CD40 ligation converts TGF-beta-secreting tolerogenic CD4-8- dendritic cells into IL-12-secreting immunogenic ones. AB - CD40L, the ligand for CD40 on dendritic cells (DCs), plays an important role in maturation and activation of DCs leading to induction of immune responses. Our previous studies showed that the mouse splenic CD4(-)8(-) DCs are tolerogenic and capable of stimulating suppressive type 1 CD4(+) regulatory T (Tr1) cell responses via TGF-beta secretion. In this study, we investigated whether CD40 ligation is able to convert tolerogenic CD4(-)8(-) DCs into immunogenic ones by in vitro treatment of DCs with anti-CD40 antibody. Our data showed that in vitro CD40 ligation with anti-CD40 antibody converted TGF-beta-secreting tolerogenic CD4(-)8(-) DCs into IL-12-secreting immunogenic ones capable of stimulating type 1 CD4(+) helper T (Th1) and CD8(+) cytotoxic T lymphocyte (CTL) responses leading to induction of antitumor immunity. In addition, in vivo CD40 ligation by intratumoral injection of adenoviral vector AdVCD40L expressing CD40 ligand also induced tumor growth inhibition and regression of established P815 tumors with infiltration of tolerogenic CD4(-)8(-) DCs. Therefore, our data provide new information for and may thus have useful impacts in CD40 ligation-based immunotherapy of cancer. PMID- 19135982 TI - Biochemical and functional characterization of the Ror2/BRIb receptor complex. AB - Ror2 belongs to the Ror family of receptor tyrosine kinases. Two distinct human disorders result from mutations in Ror2 suggesting a role in cartilage formation, chondrocyte differentiation, and joint formation. We have previously demonstrated functional and physical association of Ror2 with the BMP receptor type Ib (BRIb). The interaction site was mapped to the extracellular CRD domain of Ror2. Here we show specific association with and transphosphorylation by BRIb, but not BMP receptors Ia or II. This association is independent of N-glycosylation, excluding the possibility that the interaction is mediated by carbohydrate moieties present in the CRD region of Ror2. The Ror2/BRIb complex proved very stable under high ionic and reducing conditions, yet it appeared sensitive to SDS-treatment. Besides we provide evidence that the Ror2/BRIb complex forms in distinct microdomains at the plasma membrane (DRMs), indicating that Ror2 may interfere with BMP signaling complexes within these membrane domains. PMID- 19135983 TI - Cholinergic mechanism involved in the nociceptive modulation of dentate gyrus. AB - Acetylcholine (ACh) causes a wide variety of anti-nociceptive effects. The dentate gyrus (DG) region of the hippocampal formation (HF) has been demonstrated to be involved in nociceptive perception. However, the mechanisms underlying this anti-nociceptive role have not yet been elucidated in the cholinergic pain related neurons of DG. The electrical activities of pain-related neurons of DG were recorded by a glass microelectrode. Two kinds of pain-related neurons were found: pain-excited neurons (PEN) and pain-inhibited neurons (PIN). The experimental protocol involved intra-DG administration of muscarinic cholinergic receptor (mAChR) agonist or antagonist. Intra-DG microinjection of 1 microl of ACh (0.2 microg/microl) or 1 microl of pilocarpine (0.4 microg/microl) decreased the discharge frequency of PEN and prolonged firing latency, but increased the discharge frequency of PIN and shortened PIN inhibitory duration (ID). Intra-DG administration of 1 microl of atropine (1.0 microg/microl) showed exactly the opposite effects. According to the above experimental results, we can presume that cholinergic pain-related neurons in DG are involved in the modulation of the nociceptive response by affecting the discharge of PEN and PIN. PMID- 19135984 TI - ZIC1 is downregulated through promoter hypermethylation in gastric cancer. AB - As one of major epigenetic changes responsible for tumor suppressor gene inactivation in the development of cancer, promoter hypermethylation was proposed as a marker to define novel tumor suppressor genes. In the current study we identified ZIC1 (Zic family member 1, odd-paired Drosophila homolog) as a novel tumor suppressor gene silenced through promoter hypermethylation in gastric cancer, the second leading cause of cancer death worldwide. In all of gastric cancer cells lines examined, ZIC1 expression was downregulated and such downregulation was accompanied with the hypermethylation of ZIC1 promoter. Demethylation treatment with 5-aza-2'-deoxycytidine (Aza) reversed ZIC1 downregulation, highlighting the importance of promoter methylation to ZIC1 downregulation in gastric cancer cells. Notably, ZIC1 expression was significantly downregulated in primary gastric carcinoma tissues in comparison with non-tumor adjacent gastric tissues (p<0.01). Accordingly, promoter methylation of ZIC1 was frequently detected in primary gastric carcinoma tissues (94.6%, 35/37) but not normal gastric tissues, indicating that promoter hypermethylation mediated ZIC1 downregulation may play an important role in gastric carcinogenesis. Indeed, ectopic expression of ZIC1 led to the growth inhibition of gastric cancer cells through the induction of S-phase cell cycle arrest (p<0.01). Our results revealed ZIC1 as a novel candidate tumor suppressor gene downregulated through promoter hypermethylation in gastric cancer. PMID- 19135985 TI - Overexpression of a NAC transcription factor enhances rice drought and salt tolerance. AB - The plant-specific NAC (NAM, ATAF1/2, CUC2) transcription factors play diverse roles in plant development and stress responses. In this study, a rice NAC gene, ONAC045, was functionally characterized, especially with regard to its role in abiotic stress resistance. Expression analysis revealed that ONAC045 was induced by drought, high salt, and low temperature stresses, and abscisic acid (ABA) treatment in leaves and roots. Transcriptional activation assay in yeast indicated that ONAC045 functioned as a transcriptional activator. Transient expression of GFP-ONAC045 in onion epidermal cells revealed that ONAC045 protein was localized in the nucleus. Transgenic rice plants overexpressing ONAC045 showed enhanced tolerance to drought and salt treatments. Two stress-responsive genes were upregulated in transgenic rice. Together, these results suggest that ONAC045 encodes a novel stress-responsive NAC transcription factor and is potential useful for engineering drought and salt tolerant rice. PMID- 19135986 TI - Selective blockade of CaMKII-alpha inhibits NMDA-induced caspase-3-dependent cell death but does not arrest PARP-1 activation or loss of plasma membrane selectivity in rat retinal neurons. AB - Calcium/calmodulin-dependent protein kinase II-alpha (CaMKII-alpha) has been implicated in a number of receptor mediated events in neurons. Pharmacological blockade of CaMKII-alpha has been shown to prevent phosphorylation of NMDA-R2A and R2B receptor subunits, suggesting that this enzyme may be linked to receptor trafficking of glutamate receptors and serve as a regulatory protein for neuronal cell death. In the retina, inhibition of CaMKII-alpha has been reported to be neuroprotective against NMDA-induced cell death by preventing the activation of the caspase-3 dependent pathway. However, the effects of CaMKII-alpha blockade on the caspase-3 independent, PARP-1 dependent and the non-programmed cell death pathways have not previously been investigated. In the present study, blockade of CaMKII-alpha with the highly specific antagonist myristoylated autocamtide-2 related inhibitory peptide (AIP) was used in a rat in vivo model of retinal toxicity to compare the effects of on NMDA-induced caspase-3-dependent, PARP-1 dependent and the non-programmed (necrosis) cell death pathways. Results confirmed that AIP fully attenuates caspase-3 activation for at least 8 h following NMDA insult and also significantly improves retinal ganglion cell survival. However, this blockade had little effect on reducing the loss of plasma membrane selectivity (LPMS, e.g. necrosis) in cells located in the ganglion cell and inner nuclear layers and did not alter NMDA-induced PARP-1 hyperactivation, or prevent TUNEL labeling following a moderate NMDA-insult. These findings support a specific role for CaMKII-alpha in mediating the caspase-3 dependent cell death pathway and provide evidence that it is not directly linked to the signaling of either the PARP-1 dependent or the non-programmed cell death pathways. PMID- 19135988 TI - Increased monocyte alphaL, alphaM and beta2 integrin subunits in diabetes mellitus. AB - OBJECTIVES: Previous studies indicated the involvement of alpha2 (CD49b) integrin subunit in monocyte-laminin attachment in diabetes. The aim of the present study was to further investigate monocyte atherosclerosis related integrin subunit changes in relation to diabetes and hyperglycemia. DESIGN AND METHODS: Monocytes were derived from 10 patients with type II diabetes and 10 age matched controls. Attachment to laminin-1 in the presence of anti-integrin subunit antibodies has been estimated by a solid phase assay while for the alpha2 integrin subunit density assessment, fluorescence spectrometry was used. RESULTS: Incubation of monocytes with monoclonal anti-alphaL (CD11a) and anti-beta2 (CD18) equalized the differences between diabetics and controls. Both glucose and insulin increased the alpha2 integrin subunit expression in relation to the controls. CONCLUSIONS: The previously observed modified interaction between monocytes and laminin in diabetes mellitus may be attributed to changes not only in alpha2 but also in alphaL and beta2 integrin subunit changes. PMID- 19135987 TI - Effects of progesterone administration on infarct volume and functional deficits following permanent focal cerebral ischemia in rats. AB - Recent experimental evidence indicates that progesterone (PROG) protects against various models of brain injury, including ischemic stroke. Most human studies of pharmacologic treatments for acute cerebral stroke have failed despite initial success in animal models. To simulate better the typical human stroke without reperfusion, the present study was conducted to examine the efficacy of PROG on infarct volume and functional outcome in a permanent model of stroke, using direct cauterization of the middle cerebral artery (MCA). Twenty-four male adult Sprague-Dawley rats underwent pMCAO by electro-coagulation and sham operation. After induction of permanent MCA occlusion (pMCAO), the rats received an initial intraperitoneal injection of PROG (8 mg/kg) or vehicle at 1 h post-occlusion followed by subcutaneous injections at 6, 24 and 48 h. Functional deficits were tested on the rotarod and grip-strength meter at 24, 48 and 72 h after pMCAO. The rats were killed 72 h after surgery and isolated brain was sectioned into coronal slices and stained with 2, 3, 5-triphenyltetrazolium chloride (TTC). PROG-treated rats showed a substantial reduction (54.05%) in the volume of the infarct (% contralateral hemisphere) compared to vehicle controls. In addition there was a significant improvement in ability to remain on an accelerating rotarod and increased grip strength observed in the pMCAO rats treated with PROG compared to vehicle. Taken together, these data indicate that PROG is beneficial in one of the best-characterized models of stroke, and may warrant further testing in future clinical trials for human stroke. PMID- 19135989 TI - Reverse modulation of the HDL anionic peptide factor and phospholipid transfer protein activity in coronary artery disease and type 2 diabetes mellitus. AB - OBJECTIVES: The high density lipoprotein Anionic Peptide Factor (HDL(3)-APF) was previously described as an apolipoprotein that promotes the reverse cholesterol transport. Since phospholipid transfer protein (PLTP) is involved in such mechanism we attempted to focus on the two APF and PLTP proteins. DESIGN AND METHODS: We recruited 56 type 2 diabetic patients with (n=36) or without (n=20) coronary artery disease (CAD) and 19 CAD patients. The three groups were compared to 39 healthy control subjects. In all groups, lipid profile was determined and plasma APF concentrations and PLTP activity were measured. RESULTS: In all patients, the PLTP activity was significantly increased in comparison with controls (p<0.01), in concomitance with a plasma APF level decrease in groups with CAD (with and without type 2 diabetes) (p<0.01). Multiple linear regression analysis demonstrated that, when apoA-I, HDL-C, HDL-phospholipids and PLTP activity were taken into account as independent variables (after univariate regression analysis), HDL-PL was positively and independently related to APF (p<0.0001 in whole population; p=0.0090 in controls) and PLTP activity was negatively and independently related to APF in whole population and all patients' groups (all p<0.05), but positively and independently associated to APF in controls (p=0.0005). CONCLUSIONS: APF could be considered as a specific marker against CAD and type 2 diabetes mellitus and our results confirm the atherogenic behavior of PLTP in CAD. Thus, these two proteins are likely to be regulated in a reverse manner. PMID- 19135990 TI - Trehalose loading into red blood cells is accompanied with hemoglobin oxidation and membrane lipid peroxidation. AB - One of the recent approaches to enhance desiccation tolerance in red blood cells (RBCs) is by loading trehalose. This process has been shown to increase the recovery of lyophilized RBCs; conversely, it results in cellular damage including hemoglobin oxidation and loss of membrane integrity. The purpose of this study was to further investigate the extent of oxidative injury during the loading of trehalose into RBCs. RBCs were incubated in the absence (control) or presence of trehalose (0.8 mol/l) at 4 degrees C or 37 degrees C for different time scales. Oxidative damage was monitored by flow cytometry using dichlorofluorescin for reactive oxygen species formation, Annexin V-FITC for phosphatidylserine translocation and fluorescein-DHPE for lipid peroxidation. Percent methemoglobin, percent hemolysis and thiobarbituric acid reactive substances were measured by spectrophotometry. The extent of oxidative damage during trehalose loading is affected by the incubation temperature, incubation time and the presence of trehalose. Incubation at 4 degrees C was relatively innocuous; however, oxidative injury was evident at 37 degrees C in both RBC groups. The addition of trehalose is correlated with high osmotic pressure, which had minor effects during incubation at 4 degrees C, but seemed to have exacerbated the severity of cellular injury at 37 degrees C, as measured by higher levels of hemolysis, methemoglobin and lipid peroxidation. The process of trehalose-loading is problematic due to its requirement for prolonged incubations at 37 degrees C. These conditions are correlated with oxidative injury, even in the absence of trehalose. While trehalose is believed to be crucial for stabilizing biomembranes, the consequences of its introduction into the cells require further investigation. PMID- 19135991 TI - Cryoprotective effects of antifreeze proteins delivered into zebrafish embryos. AB - Fish embryo cryopreservation, which is useful in aquaculture or biodiversity conservation, is still far from being achieved. Structural barriers reduce the entrance of cryoprotectants into embryo compartments. Previous studies demonstrated a better ability for freezing in Arctic species which naturally express antifreeze proteins (AFPs). In this study, AFPs were delivered in early zebrafish embryos by incubation in media containing protein. Their cryoprotective effects were then analyzed. Chilling sensitivity was evaluated at 4 degrees C and -10 degrees C. Survival rates significantly increased in embryos incorporating AFPI and kept at -10 degrees C. To analyze their effects on cryopreservation, 5 somite embryos were vitrified. Incorporation of AFPI reduced the percentage of embryos that collapsed at thawing (14.2% of AFPI-treated embryos and 48.9% of controls). Cellular damage caused by vitrification was assessed after thawing by cell dissociation and further analysis of cell survival in culture (SYBR-14/IP labeling). The percentage of viable cells at thawing ranged from 25 to 50%, considered incompatible with embryo development. Cells recovered from frozen control embryos did not survive in culture. However, the incorporation of AFPs allowed survival similar to that of cells recovered from non-frozen embryos. Blastomere cryopreservation trials incorporating AFPI in the extender also demonstrated a significant increase in viability after freezing. Our findings demonstrated that delivery of AFPs into zebrafish embryos by incubation in media containing protein at early stages is a simple and harmless method that increases cryoprotection of the cellular compartment. This beneficial effect is also noticed in blastomeres, encouraging their use in further protocols for embryo cryopreservation. PMID- 19135992 TI - Identification and subclassification of new Atoh1 derived cell populations during mouse spinal cord development. AB - At spinal levels, sensory information pertaining to body positioning (proprioception) is relayed to the cerebellum by the spinocerebellar tracts (SCTs). In the past we revealed the basic helix-loop-helix transcription factor Atoh1 (Math1) to be important for establishing Dorsal Progenitor 1 (DP1) commissural interneurons, which comprise a subset of proprioceptive interneurons. Given there exists multiple subdivisions of the SCT we asked whether Atoh1 may also play a role in specifying other cell types in the spinal cord. Here, we reveal the generation of at least three DP1 derived interneuron populations that reside at spatially restricted positions along the rostral-caudal axis. Each of these cell populations expresses distinct markers and anatomically coincides with the cell bodies of the various subdivisions of the SCT. In addition, we found that as development proceeds (e.g. by E13.5) Atoh1 expression becomes apparent in the dorsal midline in the region of the roof plate (RP). Interestingly, we find that cells derived from Atoh1 expressing RP progenitors express SSEA-1, and in the absence of Atoh1 these progenitors become SOX9 positive. Altogether we reveal the existence of multiple Atoh1 dependent cell types in the spinal cord, and uncover a novel progenitor domain that arises late in development. PMID- 19135993 TI - S 35171 exerts protective effects in spontaneously hypertensive stroke-prone rats by preserving mitochondrial function. AB - S 35171 is one of a family of compounds that have been designed to protect mitochondrial function. We tested the hypothesis that S 35171 exerts protective effects in spontaneously hypertensive stroke-prone rats (SHRSPs), an animal model developing spontaneous brain damage preceded by proteinuria and systemic inflammation revealed by the urinary accumulation of acute-phase proteins (APPs) originating in the liver. Male SHRSPs fed a permissive diet received vehicle or S 35171 (10 mg/kg/day) started simultaneously with a high-sodium diet (group A) or after the establishment of proteinuria (group B). The drug delayed urinary APPs accumulation and the appearance of magnetic resonance imaging (MRI)-monitored brain lesions (after 62+/-3 days in group A, and 51+/-2 days in controls, P<0.01). The delay was more pronounced in group B as 30% of the animals survived the entire 90-day experimental period without brain abnormality. Proteomic analysis showed no significant alteration in the expression pattern of brain mitochondrial proteins, but the liver mitochondrial levels of carbamoylphosphate synthase I (CPS-I), an enzyme involved in urea metabolism) and the antioxidant peroxiredoxin-3 spot were affected by hypertension and S 35171. Stress reduces CPS-I and induces the peroxiredoxin-3 spot, whereas S 35171 brought about normal CPS-I expression and a 12-fold higher level of the peroxiredoxin-3 spot. As both enzymes are involved in maintaining mitochondrial functions, their increased expression after S 35171 treatment may be responsible for delaying the pathological condition that leads to the development of brain damage in SHRSPs. PMID- 19135994 TI - Rapid identification of Acanthamoeba from contact lens case using loop-mediated isothermal amplification method. AB - A method employing loop-mediated isothermal amplification (LAMP) of 18S ribosomal RNA gene was developed to detect Acanthamoeba in contact lens cases. A prevalence of 7% (10/150) was detected, with 100% sensitivity and 100% specificity when compared with the standard culture technique. Using visual inspection of turbidity a minimum of 10pg of Acanthamoeba DNA could be detected, 10 times more sensitive than quantitative PCR employing two of the LAMP primers. The production of LAMP amplicons was confirmed by gel-electrophoresis and ethidium bromide staining. The LAMP procedure takes less than 2h to perform and will be useful for incorporation into a point-of-care screening of suspected Acanthamoeba infection. PMID- 19135995 TI - Glibenclamide modulates glucantime activity and disposition in Leishmania major. AB - A source of chemotherapeutic failure in anti-infective therapies is the active movement of drugs across membranes, through ATP-binding cassette (ABC) transporters. In fact, simultaneous administration of therapeutic drugs with ABC transporter blockers has been invoked to be the way to actively prevent the emergence of drug resistance. Herein, we demonstrate that glucantime's efficacy in decreasing the infection rate of Leishmania-infected macrophages is strongly enhanced when used in combination with glibenclamide, a specific blocker of ABC transporters. Intracellular ABC transporters mediate glucantime sequestration in intracellular organelles. Their selective inhibition may effectively increase the cytoplasmic concentration of glucantime and its leishmanicidal activity. Our results reveal for the first time that glibenclamide targets in Leishmania major a compartment associated with a multivesicular system that is simultaneously labeled by the acidic marker LysoTracker-red and may represent the organelle where antimonials are sequestered. These results constitute a proof of concept that conclusively demonstrates the potential value that combination therapy with an ABC transporter blocker may have for leishmaniasis therapy. PMID- 19135996 TI - Adipose-derived mesenchymal stem cells alleviate experimental colitis by inhibiting inflammatory and autoimmune responses. AB - BACKGROUND & AIMS: Crohn's disease is a chronic disease characterized by severe T helper (Th)1 cell-driven inflammation of the colon partially caused by a loss of immune tolerance against mucosal antigens. Mesenchymal stem cells were recently described to suppress effector T-cell responses and have therapeutic effects in some immune disorders. Here, we investigated the potential therapeutic effects of human adipose-derived mesenchymal stem cells (hASCs) in a model of inflammatory bowel disease. METHODS: Mice with trinitrobenzene sulfonic acid-induced colitis were treated with hASCs after onset of disease and clinical scores were evaluated. Inflammatory response was determined by measuring the levels of different inflammatory mediators in colon and serum. Th1-mediated effector responses were evaluated by determining the proliferation and cytokine profile of activated mesenteric lymph node cells. The number of regulatory T cells and the suppressive capacity on Th1 cell responses was determined. RESULTS: Systemic infusion of hASCs or murine ASCs ameliorated the clinical and histopathologic severity of colitis, abrogating body weight loss, diarrhea, and inflammation and increasing survival (P < .001). This therapeutic effect was mediated by down regulating both Th1-driven autoimmune and inflammatory responses. ASCs decreased a wide panel of inflammatory cytokines and chemokines and increased interleukin 10 levels (P < .001), directly acting on activated macrophages. hASCs also impaired Th1 cell expansion and induced/activated CD4(+)CD25(+)FoxP3(+) regulatory T cells with suppressive capacity on Th1 effector responses in vitro and in vivo (P < .001). CONCLUSIONS: hASCs emerge as key regulators of immune tolerance and as attractive candidates for a cell-based therapy for Crohn's disease. PMID- 19135997 TI - A fast-track program reduces complications and length of hospital stay after open colonic surgery. AB - BACKGROUND & AIMS: A fast-track program is a multimodal approach for patients undergoing colonic surgery that combines stringent regimens of perioperative care (fluid restriction, optimized analgesia, forced mobilization, and early oral feeding) to reduce perioperative morbidity, hospital stay, and cost. We investigated the impact of a fast-track protocol on postoperative morbidity in patients after open colonic surgery. METHODS: A randomized trial of patients in 4 teaching hospitals in Switzerland included 156 patients undergoing elective open colonic surgery who were assigned to either a fast-track program or standard care. The primary end point was the 30-day complication rate. Secondary end points were severity of complications, hospital stay, and compliance with the fast-track protocol. RESULTS: The fast-track protocol significantly decreased the number of complications (16 of 76 in the fast-track group vs 37 of 75 in the standard care group; P = .0014), resulting in shorter hospital stays (median, 5 days; range, 2-30 vs 9 days, respectively; range, 6-30; P < .0001). There was a trend toward less severe complications in the fast-track group. A multiple logistic regression analysis revealed fluid administration greater than the restriction limits (odds ratio, 4.198; 95% confidence interval, 1.7-10.366; P = .002) and a nonfunctioning epidural analgesia (odds ratio, 3.365; 95% confidence interval, 1.367-8.283; P = .008) as independent predictors of postoperative complications. CONCLUSIONS: The fast-track program reduces the rate of postoperative complications and length of hospital stay and should be considered as standard care. Fluid restriction and an effective epidural analgesia are the key factors that determine outcome of the fast-track program. PMID- 19135998 TI - Improved virologic response in chronic hepatitis C genotype 4 treated with nitazoxanide, peginterferon, and ribavirin. AB - BACKGROUND & AIMS: Sustained virologic response (SVR) rates of 50%-60% have been achieved in patients with chronic hepatitis C genotype 4 treated with peginterferon plus ribavirin. The safety and efficacy of nitazoxanide plus peginterferon alfa-2a, with or without ribavirin, were evaluated in a randomized controlled trial at 2 centers in Egypt. METHODS: Previously untreated patients with chronic hepatitis C and genotype 4 infection were assigned randomly to groups that were given standard of care (peginterferon alfa-2a and ribavirin for 48 weeks, n = 40), nitazoxanide monotherapy for 12 weeks followed by nitazoxanide plus peginterferon alfa-2a for 36 weeks (n = 28), or nitazoxanide monotherapy for 12 weeks followed by nitazoxanide plus peginterferon alfa-2a and ribavirin for 36 weeks (n = 28). Therapeutics included nitazoxanide (500 mg) twice daily, peginterferon alfa-2a (180 microg) once weekly, and weight-based ribavirin (1000 1200 mg/day). RESULTS: The percentages of rapid virologic response (RVR), defined as undetectable HCV RNA at week 4 of combination therapy, and SVR were significantly higher in patients given the triple therapy compared with the standard of care (64% vs 38%, P = .048; and 79% vs 50%, P = .023; respectively). Patients given nitazoxanide plus peginterferon alfa-2a had intermediate rates of RVR (54%) and SVR (61%). Adverse events were similar across treatment groups except for higher rates of anemia in the groups receiving ribavirin. CONCLUSIONS: The combination of nitazoxanide, peginterferon alfa-2a, and ribavirin increased the percentages of patients with RVR and SVR, compared with patients given peginterferon plus ribavirin, without an increase in adverse events. PMID- 19135999 TI - The relationship between fitness and baseline glucocorticoids in a passerine bird. AB - Glucocorticoid (cort) hormones are increasingly applied in studies of free ranging animals, with elevated baseline cort levels generally assumed to indicate individuals or populations in worse condition and with lower fitness (the Cort Fitness Hypothesis). The relationship between cort and fitness is rarely validated and studies investigating the cort-fitness relationship often find results inconsistent with the Cort-Fitness Hypothesis. The inconsistency of these studies may result in part from variation in the cort-fitness relationship across life history stages. Here we address the following questions in a two-year study in free-ranging tree swallows (Tachycineta bicolor): (1) Do baseline cort levels correlate with fitness within a life history stage? (2) Does the cort-fitness relationship vary across different life history stages? (3) Does the cort-fitness relationship vary across life history stages within an individual? (4) Does reproductive effort influence cort levels, and do cort levels influence reproductive effort? We measured baseline cort and fitness components in female birds of known breeding stages. We find correlations between baseline cort levels and fitness within some life history stages, but the relationship shifts from negative during early breeding to positive during late breeding, even within the same individuals. A positive relationship between baseline cort and fitness components during the nestling period suggests that reproductive investment may elicit higher cort levels that feedback to reallocate more effort to reproduction during critical periods of nestling provisioning. Our findings provide reason to question the Cort-Fitness Hypothesis, and have implications for the application of cort measures in monitoring the condition of populations of conservation concern. PMID- 19136001 TI - Relationship between plasma leptin-like protein levels, begging and provisioning in nestling thin-billed prions Pachyptila belcheri. AB - While there have been many studies in various species examining the physiological role of leptin, there are so far no data in free-living seabirds. In the present study, we assess whether leptin is expressed in thin-billed prions (Pachyptila belcheri) and we investigate its relationship with feeding-related parameters including body condition, begging intensities and provisioning rates. We showed by Western Blot analysis using leptin-specific antibody that leptin-like protein (14-16kDa) is expressed in adipose tissue and liver of nestling thin-billed prions. Plasma leptin-like protein levels, determined by RIA, were in the same range (1-3ng/ml) as in other avian species and increased with age. In two breeding seasons, the plasma leptin-like protein levels were negatively correlated with provisioning rates (R=-0.67 and -0.35 in 2003 and 2004, respectively, P<0.05) indicating that endogenous leptin may be an anorexigenic hormone in wild birds. Plasma leptin-like protein levels were positively correlated with begging intensities (R=0.43 and 0.37 in 2003 and 2004, respectively, P<0.05), and this may be because hungry nestling seabird chicks with low body conditions increased their begging intensities. Plasma leptin-like protein levels did not correlate either with plasma triglyceride or glucose levels in thin-billed prions. Overall, these findings show the presence of leptin like protein in free-living seabirds and provide new insights into its function and its possible role in feeding-associated behaviours. PMID- 19136000 TI - Modulation of metabolic and clock gene mRNA rhythms by pineal and retinal circadian oscillators. AB - Avian circadian organization involves interactions between three neural pacemakers: the suprachiasmatic nuclei (SCN), pineal, and retina. Each of these structures is linked within a neuroendocrine loop to influence downstream processes and peripheral oscillations. However, the contribution of each structure to drive or synchronize peripheral oscillators or circadian outputs in avian species is largely unknown. To explore these interactions in the chick, we measured 2-deoxy[(14)C]-glucose (2DG) uptake and mRNA expression of the chick clock genes bmal1, cry1, and per3 in three brain areas and in two peripheral organs in chicks that underwent pinealectomy, enucleation, or sham surgery. We found that 2DG uptake rhythms damp under constant darkness in intact animals, while clock gene mRNA levels continue to cycle, demonstrating that metabolic rhythms are not directly driven by clock gene transcription. Moreover, 2DG rhythms are not phase-locked to rhythms of clock gene mRNA. However, pinealectomy and enucleation had similar disruptive effects on both metabolic and clock gene rhythms, suggesting that both of these oscillators act similarly to reinforce molecular and physiological rhythms in the chicken. Finally, we show that the relative phasing of at least one clock gene, cry1, varies between central and peripheral oscillators in a tissue specific manner. These data point to a complex, differential orchestration of central and peripheral oscillators in the chick, and, importantly, indicate a disconnect between canonical clock gene regulation and circadian control of metabolism. PMID- 19136002 TI - Trypanosoma cruzi bromodomain factor 2 (BDF2) binds to acetylated histones and is accumulated after UV irradiation. AB - Histone tail post-translational modifications (acetylation, methylation, phosphorylation, ubiquitination and ADP-ribosylation) regulate many cellular processes. Among these modifications, phosphorylation, methylation and acetylation have already been described in trypanosomatid histones. Bromodomains, together with chromodomains and histone-binding SANT domains, were proposed to be responsible for "histone code" reading. The Trypanosoma cruzi genome encodes four coding sequences (CDSs) that contain a bromodomain, named TcBDF1-4. Here we show that one of those, TcBDF2, is expressed in discrete regions inside the nucleus of all the parasite life cycle stages and binds H4 and H2A purified histones from T. cruzi. Immunolocalization experiments using both anti-histone H4 acetylated peptides and anti-TcBDF2 antibodies determined that TcBDF2 co-localizes with histone H4 acetylated at lysines K10 and K14. TcDBF2 and K10 acetylated H4 interaction was confirmed by co-immunoprecipitation. It is also shown that TcBDF2 was accumulated after UV irradiation of T. cruzi epimastigotes. These results suggest that TcBDF2 could be taking part in a chromatin remodelling complex in T. cruzi. PMID- 19136003 TI - Plasmodium male development gene-1 (mdv-1) is important for female sexual development and identifies a polarised plasma membrane during zygote development. AB - Successful development of Plasmodium sexual stages is essential for parasite survival, but the genes involved are poorly understood. We 'knocked out' the male development gene-1 (mdv-1) locus in Plasmodium berghei and found it to be important in female gametocyte activation. Indirect immunofluorescence assays show MDV-1 has a punctate cytoplasmic distribution in gametocytes. After activation of both females and males, MDV-1 is more peripherally located but in males exclusively it becomes concentrated in a few large foci. In vitro ookinete conversion assays that test the ability of activated female gametocytes to develop into retort stage ookinetes, suggests a complicit role for MDV-1, with the knock-out parasite producing 86% reduction in ookinetes. The retort stage ookinete develops from the zygote by increasing growth of an apical protrusion and MDV-1 locates at the 'leading' extracellular apical pole of this protrusion. In the fully developed ookinete MDV-1 is localised to the posterior pole. In vivo, the knock-out parasites demonstrate a phenotype in which there is a 90% reduction of parasite transmission to oocysts in mosquitoes. PMID- 19136004 TI - An apicomplexan ankyrin-repeat histone deacetylase with relatives in photosynthetic eukaryotes. AB - Cryptosporidium parvum is a member of the Apicomplexa that lacks a plastid and associated nuclear-encoded genes, which has hampered its use in evolutionary comparisons with algae and eliminated a pool of potentially useful drug targets. Here we show that apicomplexan parasites possess an unusual family of class II histone deacetylase (HDAC) proteins with orthologues that are present in other chromalveolates and primitive algae. A striking feature of these HDAC proteins is the presence of ankyrin repeats in the amino-terminus that appear to be required for enzyme activity. In vitro and in vivo analyses of the C. parvum orthologue indicate that this subclass of chromatin-remodelling proteins is targeted by the anti-cancer drug suberoylanilide hydroxamic acid and that these proteins are most likely involved in the essential process of H4 histone deacetylation that coincides with DNA replication. We propose that members of this novel class of histone deacetylase can serve as promising new targets for treatments against debilitating diseases such as cryptosporidosis, toxoplasmosis and malaria. PMID- 19136005 TI - Leishmania major CorA-like magnesium transporters play a critical role in parasite development and virulence. AB - Establishment of infection by Leishmania depends on the transformation of the invading metacyclic promastigotes into the obligatory intracellular amastigotes, and their subsequent survival in the macrophage phagolysosome, which is low in magnesium. We show that two Leishmania major proteins designated MGT1 and MGT2, which play a critical role in these processes, belong to the two-transmembrane domain (2-TM-GxN) cation transporter family and share homology with the major bacterial magnesium transporter CorA. Although both are present in the endoplasmic reticulum throughout the life cycle of the parasite, MGT1 is more highly expressed in the infectious metacyclic parasites, while MGT2 is enriched in the immature procyclic stages. The two proteins, although predicted to be structurally similar, have features that suggest different regulatory or gating mechanisms. The two proteins may also be functionally distinct, since only MGT1 complements an Escherichia coli DeltaCorA mutant. In addition, deletion of one mgt1 allele from L. major led to increased virulence, while deletion of one allele of mgt2 resulted in slower growth and total loss of virulence in vitro and in vivo. This loss of virulence may be due to an impaired transformation of the parasites into amastigotes. Deletion of both mgt1 alleles in the hemizygous MGT2 knockdown parasites reversed the growth defect and partially restored virulence. Our data indicate that the MGTs play a critical role in parasite growth, development and virulence. PMID- 19136006 TI - Crystal structures of mite allergens Der f 1 and Der p 1 reveal differences in surface-exposed residues that may influence antibody binding. AB - The group 1 mite allergens Der f 1 and Der p 1 are potent allergens excreted by Dermatophagoides farinae and Dermatophagoides pteronyssinus, respectively. The human immunoglobulin E antibody responses to the group 1 allergens show more cross-reactivity than the murine immunoglobulin G antibody responses, which are largely species specific. Here, we report the crystal structure of the mature form of Der f 1, which was isolated from its natural source, and a new high resolution structure of mature recombinant Der p 1. Unlike Der p 1, Der f 1 is monomeric both in the crystalline state and in solution. Moreover, no metal binding is observed in the structure of Der f 1 despite the fact that all amino acids involved in Ca(2+) binding in Der p 1 are completely conserved in Der f 1. Although Der p 1 and Der f 1 share an extensive sequence identity, comparison of the crystal structures of both allergens revealed structural features that could explain the differences in murine IgG and human IgE antibody responses to these allergens. There are structural differences between Der f 1 and Der p 1 that are unevenly distributed on the allergens' surfaces. This uneven spatial arrangement of conserved versus altered residues could explain both the specificity and cross reactivity of antibodies against Der f 1 and Der p 1. PMID- 19136008 TI - M phase-specific phosphorylation of histone H1.5 at threonine 10 by GSK-3. AB - H1 histones are progressively phosphorylated during the cell cycle. The number of phosphorylated sites is zero to three in late S phase and increases to five or six in late G2 phase and M phase. It is assumed that this phosphorylation modulates chromatin condensation and decondensation, but its specific role remains unclear. Recently, it was shown that the somatic H1 histone subtype H1.5 becomes pentaphosphorylated during mitosis, with phosphorylated threonine 10 being the last site to be phosphorylated. We have generated an antiserum specific for human H1.5 phosphorylated at threonine 10. Immunofluorescence labeling of HeLa cells with this antiserum revealed that the phosphorylation at this site appears in prometaphase and disappears in telophase, and that this hyperphosphorylated form of H1.5 is mainly chromatin-bound in metaphase when chromatin condensation is maximal. In search of the kinase responsible for the phosphorylation at this site, we found that threonine 10 of H1.5 can be phosphorylated by glycogen synthase kinase-3 in vitro, but not by cyclin dependent kinase 1/cyclin B and cyclin-dependent kinase 5/p35, respectively. Furthermore, addition of specific glycogen synthase kinase-3 inhibitors led to a reduction in phosphorylation at this site both in vivo and in vitro. PMID- 19136007 TI - Downhill versus barrier-limited folding of BBL 1: energetic and structural perturbation effects upon protonation of a histidine of unusually low pKa. AB - A dispersion of melting temperatures at pH5.3 for individual residues of the BBL protein domain has been adduced as evidence for barrier-free downhill folding. Other members of the peripheral subunit domain family fold cooperatively at pH7. To search for possible causes of anomalies in BBL's denaturation behavior, we measured the pH titration of individual residues by heteronuclear NMR. At 298 K, the pK(a) of His142 was close to that of free histidine at 6.47+/-0.04, while that of the more buried His166 was highly perturbed at 5.39+/-0.02. Protonation of His166 is thus energetically unfavorable and destabilizes the protein by approximately 1.5 kcal/mol. Changes in C(alpha) secondary shifts at pH5.3 showed a decrease in helicity of the C-terminus of helix 2, where His166 is located, which was accompanied by a measured decrease of 1.1+/-0.2 kcal/mol in stability from pH7 to 5.3. Protonation of His166 perturbs, therefore, the structure of BBL. Only approximately 1% of the structurally perturbed state will be present at the biologically relevant pH7.6. Experiments at pH5.3 report on a near-equal mixture of the two different native states. Further, at this pH, small changes of pH and pK(a) induced by changes in temperature will have near-maximal effects on pH dependent conformational equilibria and on propagation of experimental error. Accordingly, conventional barrier-limited folding predicts some dispersion of measured thermal unfolding curves of individual residues at pH5.3. PMID- 19136009 TI - A novel fold in the TraI relaxase-helicase c-terminal domain is essential for conjugative DNA transfer. AB - TraI relaxase-helicase is the central catalytic component of the multiprotein relaxosome complex responsible for conjugative DNA transfer (CDT) between bacterial cells. CDT is a primary mechanism for the lateral propagation of microbial genetic material, including the spread of antibiotic resistance genes. The 2.4-A resolution crystal structure of the C-terminal domain of the multifunctional Escherichia coli F (fertility) plasmid TraI protein is presented, and specific structural regions essential for CDT are identified. The crystal structure reveals a novel fold composed of a 28-residue N-terminal alpha-domain connected by a proline-rich loop to a compact alpha/beta-domain. Both the globular nature of the alpha/beta-domain and the presence as well as rigidity of the proline-rich loop are required for DNA transfer and single-stranded DNA binding. Taken together, these data establish the specific structural features of this noncatalytic domain that are essential to DNA conjugation. PMID- 19136010 TI - The chromatin of active genes is not in a permanently open conformation. AB - Quantitative measurements of local chromatin accessibility to DNase I in 15-day chicken embryo erythrocyte nuclei have been performed using a range of nuclease concentrations and real-time TaqMan PCR to monitor the loss of short ( approximately 80 bp) amplicons. At the beta-globin locus, well-established DNase I hypersensitive sites stand out against a background in which actively transcribed gene sequences (e.g., beta-adult and beta-hatching) are no more sensitive than the nearby constitutive heterochromatin that has previously been shown to form the 30-nm fibre structure. Similar observations were made at the lysozyme locus containing the active Gas41 gene and also at the GAPDH locus. We conclude that active genes are not continuously held in an open 'beads-on-a string' configuration, but adopt a 30-nm-type structure most of the time. This implies that the compact nucleosomal supercoil re-forms in the wake of the polymerase complex. PMID- 19136011 TI - Structures of dimeric GIT1 and trimeric beta-PIX and implications for GIT-PIX complex assembly. AB - GIT (G protein-coupled receptor kinase-interacting protein) and PIX (p21 activated kinase-interacting exchange factor) family proteins integrate signaling pathways involving Arf and Rho family GTPases. GIT1 and beta-PIX form a constitutively associated complex that acts as a scaffold to allow the formation of large multiprotein assemblies that regulate synaptogenesis, cell polarity and cell migration among other physiological processes. Complex formation is mediated by the GIT binding domain (GBD) in beta-PIX, which recognizes the Spa homology domain of GIT1. Both binding domains are adjacent to predicted coiled-coil segments that allow homo-oligomerization of GIT1 and beta-PIX, respectively. Oligomerization of GIT and PIX proteins is important for their physiological functions, and deletion of the coiled-coil domains interferes with correct subcellular localization and the GEF (guanine nucleotide exchange factor) activity of PIX. We have solved the crystal structures of the CC domains of GIT1 and beta-PIX and determined the stoichiometry of complex formation between the two proteins in order to understand the molecular architecture of the GIT1-beta PIX complex. The crystal structure of the CC domain of GIT1 solved at 1.4 A resolution shows a dimeric, parallel CC that spans 67 A in length. Unexpectedly, and in contrast to prevalent dimeric models, the structure of the CC region of beta-PIX determined at 2.8 A resolution, combined with hydrodynamic studies, reveals that this protein forms a parallel trimer. Furthermore, we demonstrate that dimeric GIT and trimeric PIX form an unusual high-affinity heteropentameric complex in which each Spa homology domain of the GIT1 dimer recognizes one GBD of the beta-PIX trimer, leaving one GBD unoccupied. These results can serve as a basis to better understand oligomerization-dependent GIT1-beta-PIX-regulated signaling events and provide an insight into the architecture of large signaling complexes involving GIT1 and beta-PIX. PMID- 19136012 TI - The desmoglein-specific cytoplasmic region is intrinsically disordered in solution and interacts with multiple desmosomal protein partners. AB - The desmoglein-specific cytoplasmic region (DSCR) is a conserved region of unknown structure and function that uniquely defines the desmoglein family of cell adhesion molecules. It is the site of caspase cleavage during apoptosis, and its mutation is linked to cardiomyopathy. Here, we reveal that a 276-residue DSCR construct of human desmoglein 1 is intrinsically disordered and forms an interaction hub for desmosomal proteins. In solution, it contains 6.5% helical and 10.3% beta-strand structure based on circular dichroism spectroscopy. A single monomeric state with a predominantly unfolded structure is found by size exclusion chromatography and analytical ultracentrifugation. Thermal stability assays and nuclear magnetic resonance spectroscopy reveal a nonglobular structure under a range of solution conditions. However, the introduction of detergent micelles increases structure to 18% helical and 16% beta-strand character, suggesting an inducible structure. The DSCR exhibits weak but specific interactions with plakoglobin, the plakin domain of desmoplakin, plakophilin 1, and the cytoplasmic domain of desmocollin 1. The desmoglein 1 membrane proximal region also interacts with all four DSCR ligands, strongly with plakoglobin and plakophilin and more weakly with desmoplakin and desmocollin 1. Thus, the DSCR is an intrinsically disordered functional domain with an inducible structure that, along with the membrane proximal region, forms a flexible scaffold for cytoplasmic assembly at the desmosome. PMID- 19136013 TI - Near-UV circular dichroism reveals structural transitions of vimentin subunits during intermediate filament assembly. AB - In vitro assembly of vimentin intermediate filaments (IFs) proceeds from soluble, reconstituted tetrameric complexes to mature filaments in three distinct stages: (1) within the first seconds after initiation of assembly, tetramers laterally associate into unit-length filaments (ULFs), on average 17 nm wide; (2) for the next few minutes, ULFs grow by longitudinal annealing into short, immature filaments; (3) almost concomitant with elongation, these immature filaments begin to radially compact, yielding approximately 11-nm-wide IFs at around 15 min. The near-UV CD signal of soluble tetramers exhibits two main peaks at 285 and 278 nm, which do not change during ULF formation. In contrast, the CD signal of mature IFs exhibits two major changes: (1) the 278-nm band, denoting the transition of the tyrosines from the ground state to the first vibrational mode of the excited state, is lost; (2) a red-shifted band appears at 291 nm, indicating the emergence of a new electronic species. These changes take place independently and at different time scales. The 278-nm signal disappears within the first minute of assembly, compatible with increased rigidity of the tyrosines during elongation of the ULFs. The rise of the 291-nm band has a lifetime of approximately 13 min and denotes the generation of phenolates by deprotonation of the tyrosines' hydroxyl group after they relocalize into a negatively charged environment. The appearance of such tyrosine-binding "pockets" in the assembling filaments highlights an essential part of the molecular rearrangements characterizing the later stages of the assembly process, including the radial compaction. PMID- 19136014 TI - Downhill versus barrier-limited folding of BBL 2: mechanistic insights from kinetics of folding monitored by independent tryptophan probes. AB - Barrier-free downhill folding has been proposed for the peripheral subunit binding domain BBL. To date, ultrafast kinetic experiments on BBL, which are crucial for a mechanistic understanding of folding, have been hampered by the lack of good intrinsic spectroscopic probes. Here, we present a detailed kinetic characterization of three single-point tryptophan mutants of BBL that have suitable fluorescence properties for following microsecond and nanosecond folding kinetics using temperature jump fluorescence spectroscopy. Experiments were performed at pH 7, which is optimal for stability and minimizes complications that arise from the presence of an alternative native-state conformation of BBL at lower pH. We examined the dependence of rate and equilibrium constants on concentration of denaturant and found that they follow well-established laws allowing kinetic transients to be related to events in folding and compared with equilibrium data. Logarithms of rate constants versus denaturant concentration yielded plots (chevrons) that are characteristic of barrier-limited folding for all mutants investigated, including a truncated sequence that was previously used in the proposal of downhill folding. The thermodynamic quantities calculated from the rate constants were in excellent agreement with those directly determined from equilibrium denaturation based on empirical two-state equations. We found that sequence truncation of BBL as used in studies proposing downhill folding leads to a large loss in helical content and protein stability, which were exacerbated at the low pH used in those studies. The kinetics and equilibria of folding of BBL fit to conventional barrier-limited kinetics. PMID- 19136015 TI - Consequences of domain insertion on the stability and folding mechanism of a protein. AB - SlyD, the sensitive-to-lysis protein from Escherichia coli, consists of two domains. They are not arranged successively along the protein chain, but one domain, the "insert-in-flap" (IF) domain, is inserted internally as a guest into a surface loop of the host domain, which is a prolyl isomerase of the FK506 binding protein (FKBP) type. We used SlyD as a model to elucidate how such a domain insertion affects the stability and folding mechanism of the host and the guest domain. For these studies, the two-domain protein was compared with a single-domain variant SlyDDeltaIF, SlyD* without the chaperone domain (residues 1 69 and 130-165) in which the IF domain was removed and replaced by a short loop, as present in human FKBP12. Equilibrium unfolding and folding kinetics followed an apparent two-state mechanism in the absence and in the presence of the IF domain. The inserted domain decreased, however, the stability of the host domain in the transition region and decelerated its refolding reaction by about 10-fold. This originates from the interruption of the chain connectivity by the IF domain and its inherent instability. To monitor folding processes in this domain selectively, a Trp residue was introduced as fluorescent probe. Kinetic double mixing experiments revealed that, in intact SlyD, the IF domain folds and unfolds about 1000-fold more rapidly than the FKBP domain, and that it is strongly stabilized when linked with the folded FKBP domain. The unfolding limbs of the kinetic chevrons of SlyD show a strong downward curvature. This deviation from linearity is not caused by a transition-state movement, as often assumed, but by the accumulation of a silent unfolding intermediate at high denaturant concentrations. In this kinetic intermediate, the FKBP domain is still folded, whereas the IF domain is already unfolded. PMID- 19136016 TI - Nanomechanics of native thick filaments from indirect flight muscles. AB - During flight, the wings of Drosophila melanogaster beat nearly 200 times per second. The indirect flight muscle fibers that power this movement have evolved to resist the repetitive mechanical stress that results from the 5-ms wing beat cycle at a strain amplitude of 3.5%. In order to understand how this is achieved at the sarcomere level, we have analyzed the mechanical properties of native thick filaments isolated from indirect flight muscle. Single filaments adsorbed onto a solid support were manipulated in physiological buffer using an atomic force microscope. Images taken after the manipulation revealed that segments were stretched, on average, to 150%, with a maximum at 385% extension. The lateral force-versus-displacement curve associated with each manipulation contained information about the bending and tensile properties of each filament. The bending process was dominated by shearing between myosin dimers and yielded a shear modulus between 3 and 13 MPa. Maximum tension along the stretched filaments was observed at approximately 200% extension and varied between 8 and 17 nN. Based on current models of thick filament structure, these variations can be attributed to cross-links between myosin dimers distributed along the filament. PMID- 19136018 TI - Unilateral left prosopometamorphopsia: a neuropsychological case study. AB - We describe a patient who suddenly developed prosopometamorphopsia after a childbirth; she claimed that the left half of well-known and unfamiliar faces looked distorted. Brain MR was normal, whereas SPECT showed hypoperfusion of the left infero-lateral occipital cortex. No visual recognition defects for objects or faces were present. In three matching tasks with half-faces (Experiment 1), chimeric faces (Experiment 2), or chimeric objects (Experiment 3), the patient was impaired only when she matched pairs of chimeric faces differing in their left half; the same results were obtained after 1 year. This is the first behavioural demonstration of selective chronic metamorphopsia for the left side of faces, and provides new insights for models of face processing. PMID- 19136017 TI - The preferential dopamine D3 receptor antagonist S33138 inhibits cocaine reward and cocaine-triggered relapse to drug-seeking behavior in rats. AB - We have previously reported that selective dopamine (DA) D3 receptor antagonists are effective in a number of animal models of drug addiction, but not in intravenous drug self-administration, suggesting a limited ability to modify drug reward. In the present study, we evaluated the actions ofS33138, a novel partially selective D3 receptor antagonist, in animal models relevant to drug addiction. S33138, at doses of 0.156 or 0.625 mg/kg (i.p.), attenuated cocaine enhanced brain-stimulation reward (BSR), and the highest dose tested (2.5 mg/kg) produced a significant aversive-like rightward shift in BSR rate-frequency reward functions. Further, S33138 produced biphasic effects on cocaine self administration, i.e., a moderate dose (2.5 mg/kg, p.o.) increased, while a higher dose (5 mg/kg, p.o.) inhibited, cocaine self-administration. The increase in cocaine self-administration likely reflects a compensatory response to a partial reduction in drug reward after S33138. In addition, S33138 (0.156-2.5 mg/kg, p.o.) also dose-dependently inhibited cocaine-induced reinstatement of drug seeking behavior. The reduction in cocaine-enhanced BSR and cocaine-triggered reinstatement produced by lower effective doses (e.g., 0.156 or 0.625 mg/kg) of 533138 is unlikely due to impaired locomotion, as lower effective doses of S33138 decreased neither Ymax levels in the BSR paradigm, rotarod performance, nor locomotion. However, the higher doses (2.5 or 5 mg/kg) of S33138 also significantly inhibited sucrose self-administration and rotarod performance, suggesting non-D3 receptor-mediated effects on non-drug reward and locomotion. These data suggest that lower doses of S33138 interacting essentially with D3 receptors have pharmacotherapeutic potential in treatment of cocaine addiction, while higher doses occupying D2 receptors may influence locomotion and non-drug reward. PMID- 19136020 TI - On the evolution of the timing of reproduction. AB - In this paper, we study the evolution of the per capita rate of reproduction as a function of time in the modelling framework introduced by Eskola and Geritz [Eskola, H.T.M., Geritz, S.A.H., 2007. On the mechanistic derivation of various discrete-time population models, Bull. Math. Biol. 69, 329-346]. We assume that the total number of juveniles one adult individual can produce is a finite constant, and we study how this number should be distributed during the season, when certain interaction and mortality processes are also included in the model. If aggressive interactions between the juveniles are not included in the model, evolution is simply optimizing, and the optimal reproductive strategy is always a single Dirac delta-peak within the season. If aggressive interactions between the juveniles are included, an evolutionarily stable strategy can consist of not only one or two delta-peaks, but also of continuous reproduction during the season. Using this approach, we have also derived conditions under which the classical population dynamical models of Beverton and Holt [Beverton, R.J.H., Holt, S.J., 1957. On the dynamics of exploited fish populations. Fisheries Investigations, Ser. 219], Hassell [Hassell, M.P., 1975. Density-dependence in single-species populations. J. Animal Ecology 44, 283-295] and Ricker [Ricker, W.E., 1954. Stock and recruitment. J. Fisheries Res. Board Can. 11, 559-623] are evolutionarily stable. PMID- 19136019 TI - Amygdalar opioids modulate hypothalamic melanocortin-induced anorexia. AB - We wanted to assess the possibility that opioid activity in the central amygdala (CeA) could modulate the feeding inhibition of melanocortin stimulation of the paraventricular hypothalamus (PVN). The melanocortin system is important in both the acute regulation of satiety and feeding behavior and in the integration of long-term appetite signals. Melanotan II (MTII) is a synthetic MC3R and MC4R agonist which reduces food intake when given intracerebroventricularly (ICV) and into the PVN. Tyr-D-Ala-Gly-(me) Phe-Gly-ol (DAMGO), a micro-opioid receptor agonist, increases food intake, while opioid antagonists, like naltrexone (NTX), inhibit food intake after injection into many brain sites involved in appetite regulation, including the CeA. In food-deprived male Sprague-Dawley rats, co injected intra-PVN MTII partially blocked the orexigenic effect of co-injected intra-CeA DAMGO. Intra-CeA NTX co-injected with intra-PVN MTII reduced food intake significantly more than either alone. NTX administered intra-CeA reduced c Fos-immunoreactivity (IR) in nucleus accumbens neurons significantly compared to the intra-PVN MTII treated animals, animals co-injected intra-PVN with MTII and intra-CeA with NTX animals, and control animals. Intra-PVN MTII induced c-Fos-IR in significantly more PVN neurons than observed in control animals. Intra-CeA NTX co-injected with intra-PVN MTII induced c-Fos-IR significantly in PVN neurons relative to control and intra-CeA NTX animals. Such data support the significance of opioid action within the CeA as a modulator of the feeding regulation action of melanocortins within the PVN, occurring within the context of a larger appetitive network. PMID- 19136021 TI - On parameter estimation in population models II: multi-dimensional processes and transient dynamics. AB - Recently, a computationally-efficient method was presented for calibrating a wide class of Markov processes from discrete-sampled abundance data. The method was illustrated with respect to one-dimensional processes and required the assumption of stationarity. Here we demonstrate that the approach may be directly extended to multi-dimensional processes, and two analogous computationally-efficient methods for non-stationary processes are developed. These methods are illustrated with respect to disease and population models, including application to infectious count data from an outbreak of "Russian influenza" (A/USSR/1977 H1N1) in an educational institution. The methodology is also shown to provide an efficient, simple and yet rigorous approach to calibrating disease processes with gamma-distributed infectious period. PMID- 19136022 TI - Time-course comparison of xenobiotic activators of CAR and PPARalpha in mouse liver. AB - Constitutive androstane receptor (CAR) and peroxisome proliferator activated receptor (PPAR)alpha are transcription factors known to be primary mediators of liver effects, including carcinogenesis, by phenobarbital-like compounds and peroxisome proliferators, respectively, in rodents. Many similarities exist in the phenotypes elicited by these two classes of agents in rodent liver, and we hypothesized that the initial transcriptional responses to the xenobiotic activators of CAR and PPARalpha will exhibit distinct patterns, but at later time points these biological pathways will converge. In order to capture the global transcriptional changes that result from activation of these nuclear receptors over a time-course in the mouse liver, microarray technology was used. First, differences in basal expression of liver genes between C57Bl/6J wild-type and Car null mice were examined and 14 significantly differentially expressed genes were identified. Next, mice were treated with phenobarbital (100 mg/kg by gavage for 24 h, or 0.085% w/w diet for 7 or 28 days), and liver gene expression changes with regards to both time and treatment were identified. While several pathways related to cellular proliferation and metabolism were affected by phenobarbital in wild-type mice, no significant changes in gene expression were found over time in the Car-nulls. Next, we determined commonalities and differences in the temporal response to phenobarbital and WY-14,643, a prototypical activator of PPAR alpha. Gene expression signatures from livers of wild-type mice C57Bl6/J mice treated with PB or WY-14,643 were compared. Similar pathways were affected by both compounds; however, considerable time-related differences were present. This study establishes common gene expression fingerprints of exposure to activators of CAR and PPARalpha in rodent liver and demonstrates that despite similar phenotypic changes, molecular pathways differ between classes of chemical carcinogens. PMID- 19136023 TI - Early defects in photoreceptor outer segment morphogenesis in zebrafish ift57, ift88 and ift172 Intraflagellar Transport mutants. AB - Intraflagellar Transport (IFT) refers to a highly conserved process occurring in eukaryotic ciliated structures. In vertebrate photoreceptors, IFT mediates protein trafficking to the outer segments. The IFT particle is a multi-subunit complex and mutations in many individual components causes photoreceptor defects. In zebrafish, mutations in the ift57, ift88, and ift172 genes result in retinal degeneration by 5 days post fertilization (dpf). Although the effects of these mutations on photoreceptor survival have been described, early developmental morphogenesis remains poorly understood. We used transmission electron microscopy and immunohistochemistry to examine these mutants at 60, 72, and 96h post fertilization (hpf) and describe early photoreceptor morphogenesis defects. PMID- 19136024 TI - Neurobiology of the development of motivated behaviors in adolescence: a window into a neural systems model. AB - Adaptive motivated behaviors are at the core of a successful life. Conversely, perturbed motivated behaviors are the hallmark of psychiatric disorders. Based on the notion that most psychopathology is developmental in nature, understanding the neural mechanisms that control motivated behavior across development and in psychopathology is a critical step for preventing and treating psychiatric diseases. This review focuses on adolescence, which is the critical developmental period that determines the successful passage into adulthood. We first present a heuristic neural systems model of motivated behavior (triadic model) that integrates neuroscience theories and the emerging body of functional neuroimaging work on the neurodevelopment of motivated behavior. As a key feature of adolescence, social reorientation is particularly emphasized through the presentation of a parallel model of social integration processing network. Although not yet integrated in the triadic model, pubertal changes and their possible contribution to adolescent motivated behavior are reviewed. Similarly, given its central role in motivated actions, the dopamine system is discussed from the perspective of animal studies dedicated to changes of this system across adolescence. This review reveals vast gaps in knowledge about the neurobiology of motivated behavior in normally developing individuals, which makes the translation to psychopathology only tentative. However, it provides clear directions for future research. PMID- 19136025 TI - Revising the senior walking environmental assessment tool. AB - BACKGROUND: The Senior Walking Environmental Assessment Tool (SWEAT), an instrument for measuring built environmental features associated with physical activity of older adults, was revised to create an easier-to-use tool for use by practitioners and community members. METHODS: Inter-rater and intra-rater reliability of the modified instrument (SWEAT-R) was assessed in Portland, Oregon in 2007. Five trained observers audited street segments in 12 neighborhoods, resulting in 361 pairs of audits, including 63 repeated audits. RESULTS: Overall, 88% and 75% of items assessed had good or excellent inter-rater and intra-rater reliability, respectively. The revised instrument required less time to complete than the original instrument, while obtaining more information. CONCLUSION: SWEAT R provides easy to gather, reliable data for use in community-based audits of built environment in relation to walking among older adults. PMID- 19136026 TI - The impact of single and dual hydrothermal modifications on the molecular structure and physicochemical properties of normal corn starch. AB - Effect of single and dual hydrothermal modifications with annealing (ANN) and heat-moisture treatment (HMT) on molecular structure and physicochemical properties of corn starch was investigated. Normal corn starch was modified by ANN at 70% moisture at 50 degrees C for 24h and HMT at 30% moisture at 120 degrees C for 24h as well as by the combination of ANN and HMT. The apparent amylose content and swelling factor (SF) decreased on ANN and HMT, but amylose leaching (AML) increased. These changes were more pronounced on dual modification. The crystallinity (determined by X-ray diffraction), the gelatinization enthalpy (determined by differential scanning calorimetry) and ratio of 1047 cm(-1)/1022 cm(-1) (determined by Fourier transform infrared spectroscopy) slightly increased on ANN and decreased on HMT. The ANN and subsequent HMT (ANN-HMT) resulted in the lowest crystallinity, gelatinization enthalpy and ratio of 1047 cm(-1)/1022 cm(-1). The gelatinization temperature range decreased on ANN but increased on HMT. However, the gelatinization range of dually modified starches (ANN-HMT and HMT-ANN) was between ANN starch and HMT starch. Birefringence remained unchanged on ANN but slightly decreased on HMT as well as dual modification. Average chain length and amount of longer branch chains (DP> or =37) remained almost unchanged on ANN but decreased on HMT and dual modifications (ANN-HMT and HMT-ANN). HMT and dual modifications resulted in highly reduced pasting viscosity. ANN and HMT as well as dual modifications increased RDS content and decreased SDS and RS content. PMID- 19136028 TI - A pragmatic approach to biochemical systems theory applied to an alpha-synuclein based model of Parkinson's disease. AB - This paper presents a detailed systems model of Parkinson's disease (PD), developed utilizing a pragmatic application of biochemical systems theory (BST) intended to assist experimentalists in the study of system behavior. This approach utilizes relative values as a reasonable initial estimate for BST and provides a theoretical means of applying numerical solutions to qualitative and semi-quantitative understandings of cellular pathways and mechanisms. The approach allows for the simulation of human disease through its ability to organize and integrate existing information about metabolic pathways without having a full quantitative description of those pathways, so that hypotheses about individual processes may be tested in a systems environment. Incorporating this method, the PD model describes alpha-synuclein aggregation as mediated by dopamine metabolism, the ubiquitin-proteasome system, and lysosomal degradation, allowing for the examination of dynamic pathway interactions and the evaluation of possible toxic mechanisms in the aggregation process. Four system perturbations: elevated alpha-synuclein aggregation, impaired dopamine packaging, increased neurotoxins, and alpha-synuclein overexpression, were analyzed for correlation to qualitative PD system hypotheses present in the literature, with the model demonstrating a high level of agreement with these hypotheses. Additionally, various PD treatment methods, including levadopa and monoamine oxidase inhibition (MAOI) therapy, were applied to the disease models to examine their effects on the system. Future additions and refinements to the model may further the understanding of the emergent behaviors of the disease, helping in the identification of system sensitivities and possible therapeutic targets. PMID- 19136027 TI - GDNF--a potential target to treat addiction. AB - The glial cell line-derived neurotrophic factor (GDNF) is a secreted protein, best known for its role in the development of the central and peripheral nervous systems and the survival of adult dopaminergic neurons. More recently, accumulating evidence suggests that GDNF plays a unique role in negatively regulating the actions of drugs of abuse. In this article, we review these data and highlight the possibility that the GDNF pathway may be a promising target for the treatment of addiction. PMID- 19136029 TI - Differentiation of Cherry leaf roll virus isolates from various host plants by immunocapture-reverse transcription-polymerase chain reaction-restriction fragment length polymorphism according to phylogenetic relations. AB - A restriction fragment length polymorphism assay (RFLP) was developed to differentiate Cherry leaf roll virus (CLRV) isolates according to phylogenetic clades by examining restriction patterns from partial 3' non-coding region (NCR) genomic fragments (approx. 420bp). The 3' NCR fragment from 43 CLRV isolates belonging to different phylogenetic groups were compared after restriction analysis with the endonucleases Bsp143I, AluI, RsaI, EcoRI and Eco130I, and another 23 isolates were analyzed by computer assisted restriction analysis. The restriction endonucleases Bsp143I, AluI and RsaI enabled the differentiation of isolates from group B and all but two isolates belonging to group A. A major proportion of group E isolates could also be discriminated. The remainder of the group E isolates were indistinguishable from isolates belonging to phylogenetic group C or D2. Isolates belonging to group D1 could not be differentiated from two group A isolates. The method was applied successfully in an IC-RT-PCR-RFLP assay to differentiate samples from walnut, black elderberry and birch and determine their phylogenetic relationships. In future, this method will facilitate rapid phylogenetic classification of CLRV isolates detected in certain host plants by the universal immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR), and will be suitable for studying CLRV population diversity as well as genetic drift within virus populations. PMID- 19136030 TI - Involvement of serotoninergic 5-HT1A/2A, alpha-adrenergic and dopaminergic D1 receptors in St. John's wort-induced prepulse inhibition deficit: a possible role of hyperforin. AB - Prepulse inhibition (PPI) of acoustic startle response is a valuable paradigm for sensorimotor gating processes. Previous research showed that acute administration of St. John's wort extract (500 mg/kg, p.o.) to rats caused significant disruption of PPI while elevating monoamines levels in some brain areas. The cause-effect relationship between extract-induced PPI disruption and augmented monoaminergic transmission was studied using different serotoninergic, adrenergic and dopaminergic antagonists. The effects of hypericin and hyperforin, as the main active constituents of the extract, on PPI response were also tested. PPI disruption was prevented after blocking the serotoninergic 5-HT1A and 5-HT2A, alpha-adrenergic and dopaminergic D1 receptors. Results also demonstrated a significant PPI deficit after acute treatment of rats with hyperforin, and not hypericin. In some conditions manifesting disrupted PPI response, apoptosis coexists. Electrophoresis of DNA isolated from brains of hyperforin-treated animals revealed absence of any abnormal DNA fragmentation patterns. It is concluded that serotoninergic 5-HT1A and 5-HT2A, alpha-adrenergic and dopaminergic D1 receptors are involved in the disruptive effect of St. John's wort extract on PPI response in rats. We can also conclude that hyperforin, and not hypericin, is one of the active ingredients responsible for St. John's wort induced PPI disruption with no relation to apoptotic processes. PMID- 19136031 TI - Immune involvement in schizophrenia and autism: etiology, pathology and animal models. AB - There is increasing evidence of immune involvement in both schizophrenia and autism. Of particular interest are striking abnormalities in the expression of immune-related molecules such as cytokines in the brain and cerebral spinal fluid (CSF). It is proposed that this represents a permanent state of brain immune dysregulation, which begins during early development. One possibility is that maternal infection, a known risk factor for schizophrenia and autism, sets this immune activation in motion. Several animal models are being used to investigate this hypothesis. There is also recent evidence that, among schizophrenic subjects, those associated with maternal infection display a distinctive pathology, which suggests that diverse causes for this disorder may explain some of its heterogeneity. The human and animal results related to immune involvement suggest novel therapeutic avenues based on immune interventions. PMID- 19136032 TI - Role of CREB in vasoactive intestinal peptide-mediated wound healing in human bronchial epithelial cells. AB - Vasoactive intestinal peptide (VIP) is one of the most important sensory neuropeptides in respiratory system. We previously reported that VIP enhances wound healing and proliferation of human bronchial epithelial cells (HBECs), and these effects are mediated by intracellular signaling molecules such as protein kinase A (PKA), protein kinase C (PKC), Calmodulin (CaM), and extracellular signal-regulated kinase (ERK). In the present study, we further investigated the role of cAMP Response Element Binding Protein (CREB) in VIP-promoted protective effects in mechanical-damaged HBECs. VIP-mediated wound healing and cell proliferation in HBECs was inhibited by CREB antisense oligonucleotides (ASO) in a time-dependent manner. VIP increased the CREB DNA binding activity and expression of the p-CREB that were inhibited by VIP receptor antagonist. Increased CREB DNA binding activity and expression of the p-CREB were also partially inhibited by PKA and ERK inhibitors. These results suggest that the VIP mediated wound repair of HBECs is associated with activation of CREB via PKA and ERK dependent pathway. PMID- 19136033 TI - Ear mesenchymal stem cells: an efficient adult multipotent cell population fit for rapid and scalable expansion. AB - Bone marrow mesenchymal stem cells (BM-MSCs) have the potential to be used for tissue engineering. Nevertheless, they exhibit a low growth rate that limits their availability. In this work we use an alternative model of MSCs from the outer ear (ear mesenchymal stem cells, E-MSCs). These cells bear the characteristics of progenitor cells because of their ability to be differentiated into the three lineages of chondrocytes, osteocytes and adipocytes. This model cell population had a threefold higher cell growth rate compared to BM-MSCs. This allowed rapid testing of the scalability in microcarrier culture using bead-to bead transfer and also enabled their expansion in a 1-l bioreactor. The cells were able to maintain their potential for differentiation into the above three lineages. Therefore, E-MSCs appear to be an attractive model for assessing a number of bioengineering parameters that may affect the behavior of adult stem cells in culture. PMID- 19136034 TI - Prediction of drug release from ethylcellulose coated pellets. AB - The aim of this study was to elucidate the underlying drug release mechanisms in pellets coated with aqueous ethylcellulose dispersion, providing long term stable drug release profiles and containing different types of starter cores. The systems were thoroughly characterized using mechanical analysis; the sensitivity of drug release to the osmolality of the release medium was measured; scanning electron microscopy and optical macroscopy were used to monitor the pellets' morphology and dimensions upon exposure to different media, and drug release was measured from single and ensembles of pellets as well as from thin, free films. All experimental results indicate that diltiazem HCl release from pellets coated with ethylcellulose containing small amounts of poly(vinyl alcohol)-poly(ethylene glycol) graft copolymer is primarily controlled by drug diffusion through the intact polymeric membranes, irrespective of the type of starter core (consisting of microcrystalline cellulose or sugar, optionally coated with ethylcellulose). Importantly, the apparent diffusion coefficient of the drug in the macromolecular networks could easily be determined with thin free films and successfully be used to quantitatively predict the release rate from coated pellets. Thus, based on this knowledge and using the presented mathematical theories the development of new/ optimization of existing controlled drug delivery systems of this type can be significantly facilitated. PMID- 19136035 TI - Effects of energy density and portion size on development of acquired flavour liking and learned satiety. AB - The concept of learned satiety (LS) suggests that associations between the sensory quality and post-ingestive effects of foods may lead to acquired control of meal-size. Although a recent study appeared to support LS since participants learned to eat more of a flavoured cereal with lower energy density (ED) after repeated experience, suggesting that they adjusted voluntary intake to ensure adequate energy was consumed, the large serving portion used in training may have lead to over-satiation. To investigate this further, groups of 12 men were assigned to one of four conditions based on the trained serving portion (150 or 300 g) and presence or absence of cues to differentiate high and low ED versions. In the absence of sensory cues, neither mass consumed nor rated pleasantness differed between high and low ED conditions either before or after training, resulting in greater energy intake in the high ED condition. When sensory cues differentiated ED, intake increased significantly post-training in both the high ED condition trained with the small portion and low ED condition trained with the large portion, and flavour pleasantness changed similarly. Moreover hunger increased significantly after the food was tasted in both conditions where intake increased. These data provide further evidence that learning can moderate meal size dependent on energy content, but suggest that these changes are driven by changes in flavour liking rather than LS. PMID- 19136036 TI - Effects of adrenomedullin on systolic and diastolic myocardial function. AB - Adrenomedullin (AM) effects were studied in rabbit papillary muscles by adding increasing concentrations (10(-10) to 10(-6)M) either alone or after pre treatment with l-NNA, indomethacin, AM22-52 (AM receptor antagonist), CGRP(8-37) (CGRP receptors antagonist), KT5720 (PKA inhibitor), as well as after endocardial endothelium (EE) removal. Passive length-tension relations were constructed before and after a single concentration of AM (10(-6)M). AM concentration dependently induced negative inotropic and lusitropic effects, and increased resting muscle length (RL). At 10(-6)M, AT, dT/dt(max) and dT/dt(min) decreased 20.9+/-4.9%, 18.3+/-7.3% and 16.7+/-7.8%, respectively, and RL increased to 1.010+/-0.004L/L(max). Correcting RL to its initial value resulted in a 26.6+/ 6.4% decrease of resting tension, indicating decreased muscle stiffness, also patent in the down and rightward shift of the passive length-tension relation. The negative inotropic effect of AM was dependent on its receptor, CGRP receptor, PKA, the EE and NO, while the effects of AM on myocardial stiffness were abolished by EE damage and NO inhibition. This latter effect represents a novel mechanism of acute neurohumoral modulation of diastolic function, suggesting that AM is an important regulator of cardiac filling. PMID- 19136037 TI - Association of CTLA-4 polymorphisms and clinical-immunologic characteristics at onset of type 1 diabetes mellitus in children. AB - CTLA-4 is a homeostatic regulator of T cell activation and is believed to play a critical role in immune tolerance. Polymorphisms in the CTLA-4 promoter (-318C/T) and in exon 1 (+49 A/G) were analyzed in 300 Chilean patients with type 1 diabetes mellitus (T1D) and 310 healthy children by polymerase chain reaction restriction fragment length polymorphism. The effect of CTLA-4 allele and haplotype frequencies on the interferon-gamma, tumor necrosis factor-alpha, and transforming growth factor-beta(1) levels and the presence in serum of GAD65 and IA-2 autoantibodies at the onset of T1D was evaluated. The distribution of the CTLA-4 allele and genotype frequencies was found to be similar in patients and control children. However, among the T1D patients' carriers of GG genotype on CTLA-4 gene a higher frequency of anti-GAD65 autoantibodies (87.2%) was observed. On the other hand, higher ketoacidosis at onset, younger age at onset, and higher levels of tumor necrosis factor-alpha and interferon-gamma were observed in T1D patients carriers of G allele in comparison with the carriers of AA genotype. In conclusion, the result of this study showed that CTLA-4 +49 A/G and -318 C/T polymorphisms were not linked with a higher genetic risk for T1D. However, the presence of a GG genotype or G allele dosage was associated with a younger age of onset, higher prevalence of ketoacidosis at the moment of diagnosis and positive anti-GAD65 serum autoantibodies. PMID- 19136038 TI - Polymorphism of the TGFB1 gene is not associated with bronchial allergic asthma in a Polish population. AB - Allergic asthma is a complex genetic disorder that involves interactions between genetic and environmental factors. Some studies have indicated that transforming growth factor beta(1) (TGF-beta(1)), a pleiotropic cytokine regulating inflammatory reactions and airway remodeling, may participate in the pathogenesis of asthma. Several polymorphisms have been described in the TGFB1 gene; some were tested in allergic asthma, with conflicting results. The aim of this study was to investigate the possible associations of four TGFB1 gene polymorphisms (-800G>A, 509C>T, 869T>C, and 915G>C) with allergic asthma in a Polish population. These four single nucleotide polymorphisms were genotyped in 247 asthmatic patients (including 207 atopic individuals) and 287 unrelated healthy volunteers by means of the polymerase chain reaction-restriction fragment length polymorphism method. No significant differences between patients and controls in allele, genotype, and haplotype frequencies were reported. Logistic regression analysis of genotype distribution and allele positivity adjusted for age and sex did not reveal any significant differences between all patients or patients selected for atopy and controls. Thus, no evidence was reported for a contribution of the TGFB1 gene to allergic asthma in a Polish population. The results are discussed in the context of similar studies in other populations. PMID- 19136039 TI - Protective immunization with homologous and heterologous antigens against Helicobacter suis challenge in a mouse model. AB - Helicobacter (H.) suis colonizes the stomach of more than 60% of slaughter pigs and is also of zoonotic importance. Recently, this bacterium was isolated in vitro, enabling the use of pure cultures for research purposes. In this study, mice were immunized intranasally or subcutaneously with whole bacterial cell lysate of H. suis or the closely related species H. bizzozeronii and H. cynogastricus, and subsequently challenged with H. suis. Control groups consisted of non-immunized and non-challenged mice (negative control group), as well as of sham-immunized mice that were inoculated with H. suis (positive control group). Urease tests on stomach tissue samples at 7 weeks after challenge infection were negative in all negative control mice, all intranasally immunized mice except one, and in all and 3 out of 5 animals of the H. cynogastricus and H. suis subcutaneously immunized groups, respectively. H. suis DNA was detected by PCR in the stomach of all positive control animals and all subcutaneously immunized/challenged animals. All negative control animals and some intranasally immunized/challenged mice were PCR-negative. In conclusion, immunization using antigens derived from the same or closely related bacterial species suppressed gastric colonization with H. suis, but complete protection was only achieved in a minority of animals following intranasal immunization. PMID- 19136040 TI - The mucosal adjuvanticity of the oligodeoxynucleotides containing a non methylated CpG motif on BCG and diphtheria toxoid. AB - CpG-DNA is currently attracting attention as an effective and safe vaccine adjuvant to prevent from microbial infections. In this report, we examined the effects of oligo B, which is a synthetic CpG-DNA, in mucosal administration of Bacillus Calmette-Guerin (BCG) and diphtheria toxoid (DT). Co-administration with oligo B enhanced BCG-induced delayed type hypersensitivity to purified protein derivative (PPD) in guinea pigs. The titers of anti-DT serum IgG, IgA and mucosal IgA antibodies induced by intranasal administration with DT plus oligo B were significantly higher than that with DT alone. In both C57BL/6 and BALB/c mice, intranasal administration of DT with oligo B induced enough level of antibodies to prevent onset of diphtheria. The analysis of antibody subclasses showed that intranasal administration of oligo B induced not only IgG1 but also IgG2a, IgG2c and IgA anti-DT antibodies. In contrast, there was no or little production of the anti-DT serum IgE. Taken together our data suggest that oligo B is a powerful adjuvant in mucosal immunization. PMID- 19136041 TI - Reduced spontaneous facial mimicry in women with autistic traits. AB - Deficits in empathizing and perspective taking are defining characteristics of autism-spectrum disorders. Converging evidence suggests that these socio emotional abilities are rooted in basic mechanisms that subserve imitative behavior, and may vary with autistic traits across the population as a whole. We investigated this notion by assessing spontaneous and instructed mimicry of facial expressions in healthy male and female volunteers scoring extremely high or low on the autism-spectrum quotient questionnaire. Mimicry was recorded using electromyography of the corrugator supercilii and zygomaticus major. Results show that spontaneous mimicry in the corrugator supercilii was strongest in female participants with low AQ-scores. Mimicry in the zygomaticus major, and in the instructed facial mimicry condition, did not differ between groups. These findings indicate that the degree to which individuals exhibit spontaneous mimicry may vary as a function of both gender and autistic traits. PMID- 19136042 TI - Postural instability in cerebellar ataxia: correlations of knee, arm and trunk movements to center of mass velocity. AB - The aim of this study was to investigate the correlations between body segment movements and center of mass (COM) velocity during pathological balance corrections of spinocerebellar ataxia (SCA) patients compared with controls, and to relate correlations indicating instability to EMG activity differences. Eighteen SCA patients and 21 age-matched controls were tested. Upright standing was perturbed using rotations of the support surface. We recorded body motion and surface EMG. For lateral perturbations peaks in COM lateral velocity were larger in SCA patients than controls. These peaks were correlated with increased ("hypermetric") trunk roll downhill and reduced uphill knee flexion velocity. Subsequent arm abduction partially corrected the lateral instability. Early balance correcting responses in knee and paraspinal muscles showed reduced amplitudes compared with normal responses. Later responses were consistent with compensation mechanisms for the lateral instability created by the stiffened knee and pelvis. We conclude that truncal hypermetria coupled with insufficient uphill knee flexion is the primary cause of lateral instability in SCA patients. Holding the knees and pelvis more rigid possibly permits a reduction in the controlled degrees of freedom and concentration on arm abduction to improve lateral instability. For backwards perturbations excessive posterior COM velocity coincided with marked trunk hypermetric flexion forwards. We concluded that this flexion and the ensuing backwards shift of the pelvis result from rigidity which jeopardizes posterior stability. Timing considerations and the lack of confirmatory changes in amplitudes of EMG activity suggest that lateral and posterior instability in SCA is primarily a biomechanical response to pelvis and knee rigidity resulting from increased muscle background activity rather than changed evoked responses. PMID- 19136044 TI - Distribution of calcium channel Ca(V)1.3 immunoreactivity in the rat spinal cord and brain stem. AB - The function of local networks in the CNS depends upon both the connectivity between neurons and their intrinsic properties. An intrinsic property of spinal motoneurons is the presence of persistent inward currents (PICs), which are mediated by non-inactivating calcium (mainly Ca(V)1.3) and/or sodium channels and serve to amplify neuronal input signals. It is of fundamental importance for the prediction of network function to determine the distribution of neurons possessing the ion channels that produce PICs. Although the distribution pattern of Ca(V)1.3 immunoreactivity (Ca(V)1.3-IR) has been studied in some specific central nervous regions in some species, so far no systematic investigations have been performed in both the rat spinal cord and brain stem. In the present study this issue was investigated by immunohistochemistry. The results indicated that the Ca(V)1.3-IR neurons were widely distributed across different parts of the spinal cord and the brain stem although with variable labeling intensities. In the spinal gray matter large neurons in the ventral horn (presumably motoneurons) tended to display higher levels of immunoreactivity than smaller neurons in the dorsal horn. In the white matter, a subset of glial cells labeled by an oligodendrocyte marker was also Ca(V)1.3-positive. In the brain stem, neurons in the motor nuclei appeared to have higher levels of immunoreactivity than those in the sensory nuclei. Moreover, a number of nuclei containing monoaminergic cells, for example the locus coeruleus, were also strongly immunoreactive. Ca(V)1.3-IR was consistently detected in the neuronal perikarya regardless of the neuronal type. However, in the large neurons in the spinal ventral horn and the cranial motor nuclei the Ca(V)1.3-IR was clearly detectable in first and second order dendrites. These results indicate that in the rat spinal cord and brain stem Ca(V)1.3 is probably a common calcium channel used by many kinds of neurons to facilitate the neuronal information processing via certain intracellular mechanisms, for instance, PICs. PMID- 19136043 TI - Effects of Parkinson's disease on proprioceptive control of posture and reaching while standing. AB - Although previous studies have shown pointing errors and abnormal multijoint coordination in seated subjects with Parkinson's disease (PD) who cannot view their arm, the extent to which subjects with PD have problems using proprioception to coordinate equilibrium maintenance and goal-oriented task execution has not been adequately investigated. If a common motor program controls voluntary arm pointing movements and the accompanying postural adjustments, then impairments of proprioceptive integration in subjects with PD should have similar effects on pointing and body center of mass (CoM) control with eyes closed. Ten standing subjects with PD (OFF-medication) and 10 age matched control (CTR) subjects pointed to a target with their eyes closed and open. Although pointing accuracy was not significantly different between groups, body CoM displacements were reduced in subjects with PD, but not in CTR, when eyes were closed. In addition, with eyes closed, PD subjects showed reduced temporal coupling between pointing and CoM velocity profiles and reduced spatial coupling between pointing and CoM endpoints. This poor coupling with eyes closed could be related to the PD subjects' increased jerkiness of CoM displacements. The different effects of eye closure between CTR and PD subjects on the CoM displacements, but not pointing accuracy, are consistent with separate motor programs for the pointing and postural components of this task. Furthermore, the decoupling between the two movement components in subjects with PD when they could not use vision, suggests that the basal ganglia are involved in the integration of proprioceptive information for posture-movement coordination. PMID- 19136045 TI - Olfactory bulbectomy induces rapid and stable changes in basal and stress-induced locomotor activity, heart rate and body temperature responses in the home cage. AB - BACKGROUND: Olfactory bulbectomy (OBX) in rats causes several behavioral and neurochemical changes. However, the extent and onset of physiological and behavioral changes induced after bulbectomy have been little examined. METHODS: Male Sprague-Dawley rats received telemetric implants. Before and immediately after OBX surgery, basal and stress-induced heart rate, body temperature, and locomotor activity were measured in the home cage in sham (n=9) and OBX animals (n=11). Stress was induced using novel cage stress or witness stress. RESULTS: Bulbectomized animals differed physiologically and behaviorally from shams. Nocturnally, OBX animals were significantly more active compared with shams, had a higher core body temperature and displayed a decreased heart rate variability. During the light period, OBX animals had a significantly lower basal heart rate and a reduced heart rate variability. These effects became apparent 2-3 days after OBX surgery, and were stable over time. After witness stress, OBX animals showed smaller autonomic (body temperature and heart rate) responses compared with shams, but showed no difference in locomotor responses. In contrast, novel cage stress led to increased locomotor responses in OBX rats compared with sham rats, while no differences were found in autonomic responses. CONCLUSION: Removal of the olfactory bulbs results in rapid, stable and persistent changes in basal locomotor activity, body temperature, heart rate and heart rate variability. Although the sleep-wake cycle of these parameters is not altered, increases in circadian amplitude are apparent within 3 days after surgery. This indicates that physiological changes in the OBX rat are the immediate result of olfactory bulb removal. Further, stress responsivity in OBX rats depends on stressor intensity. Bulbectomized rats display smaller temperature and heart rate responses to less intense witness stress compared with sham rats. Increased locomotor responses to more intense novel cage stress are present in the home cage as well as the open field. The present study shows that olfactory bulbectomy has rapid and persistent influence on basal and stress-induced physiological parameters. PMID- 19136047 TI - Low osmolar contrast medium induces cellular injury and disruption of calcium homeostasis in rat glomerular endothelial cells in vitro. AB - The mechanisms of contrast medium (CM)-induced renal impairment at cellular level are not fully understood. The purpose of this study was to investigate the toxic effects of non-ionic low osmolar contrast medium (LOCM) on glomerular endothelial cells (GECs). Ioversol, the most representative LOCM used in clinic, was chosen to act on primary cultured rat GECs. When rat GECs were treated with various amount of ioversol, a dose-dependent reduction in cell viability was observed by tetrazolium dye (MTT) assay. After exposure to ioversol at dose of 100 microl/ml for 24h, nuclear condensation, nuclear fragmentation, cell apoptosis and necrosis were found in GECs. The intracellular free Ca(2+) concentrations ([Ca(2+)]i) detected by confocal laser scanning were markedly elevated in GECs treated with ioversol. The [Ca(2+)]i increase, LDH release and expression changes of apoptotic associated proteins in GECs induced by ioversol could be reversed by pretreatment with intracellular Ca(2+) chelator, 1,2-bis (o-aminophenoxy) ethane-N,N,N',N' tetra-acetic acid (BAPTA/AM). These results suggest that LOCM can decrease cell viability and cause cell injury of GECs. The elevation of [Ca(2+)]i released from intracellular calcium stores likely contribute to the LOCM-induced cell injury of GECs. PMID- 19136046 TI - Soman induces ictogenesis in the amygdala and interictal activity in the hippocampus that are blocked by a GluR5 kainate receptor antagonist in vitro. AB - Exposure to organophosphorus nerve agents induces brain seizures, which can cause profound brain damage resulting in death or long-term cognitive deficits. The amygdala and the hippocampus are two of the most seizure-prone brain structures, but their relative contribution to the generation of seizures after nerve agent exposure is unclear. Here, we report that application of 1 muM soman for 30 min, in rat coronal brain slices containing both the hippocampus and the amygdala, produces prolonged synchronous neuronal discharges (10-40 s duration, 1.5-5 min interval of occurrence) resembling ictal activity in the basolateral nucleus of the amygdala (BLA), but only interictal-like activity ("spikes" of 100-250 ms duration; 2-5 s interval) in the pyramidal cell layer of the CA1 hippocampal area. BLA ictal- and CA1 interictal-like activity were synaptically driven, as they were blocked by the AMPA/kainate receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione. As the expression of the GluR5 subunit of kainate receptors is high in the amygdala, and kainate receptors containing this subunit (GluR5KRs) play an important role in the regulation of neuronal excitability in both the amygdala and the hippocampus, we tested the efficacy of a GluR5KR antagonist against the epileptiform activity induced by soman. The GluR5KR antagonist UBP302 reduced the amplitude of the hippocampal interictal-like spikes, and eliminated the seizure-like discharges in the BLA, or reduced their duration and frequency, with no significant effect on the evoked field potentials. This is the first study reporting in vitro ictal-like activity in response to a nerve agent. Our findings, along with previous literature, suggest that the amygdala may play a more important role than the hippocampus in the generation of seizures following soman exposure, and provide the first evidence that GluR5KR antagonists may be an effective treatment against nerve agent induced seizures. PMID- 19136048 TI - Oxidative stress mediated cytotoxicity of cyanide in LLC-MK2 cells and its attenuation by alpha-ketoglutarate and N-acetyl cysteine. AB - Cyanide is a rapidly acting mitochondrial poison that inhibits cellular respiration and energy metabolism leading to histotoxic hypoxia followed by cell death. Cyanide is predominantly a neurotoxin but its toxic manifestations in non neuronal cells are also documented. This study addresses the oxidative stress mediated cytotoxicity of cyanide in Rhesus monkey kidney epithelial cells (LLC MK2). Cells were treated with various concentrations of potassium cyanide (KCN) for different time intervals and cytotoxicity was evidenced by increased leakage of intracellular lactate dehydrogenase, mitochondrial dysfunction (MTT assay) and depleted energy status of cells (ATP assay). Cytotoxicity was accompanied by lipid peroxidation indicated by elevated levels of malondialdehyde (MDA), reactive oxygen species (ROS) and reactive nitrogen species (RNS) (DCF-DA staining), diminished cellular antioxidant status (reduced glutathione (GSH), glutathione peroxidase, superoxide dismutase and catalase). These cascading events triggered an apoptotic kind of cell death characterized by oligonucleosomal DNA fragmentation and nuclear fragmentation (Hoechst 33342 staining). Apoptosis was further confirmed by increased caspase-3 activity. Cyanide-induced cytotoxicity, oxidative stress, and DNA fragmentation were prevented by alpha-ketoglutarate (A-KG) and N-acetyl cysteine (NAC). A-KG is a potential cyanide antidote that confers protection by interacting with cyanide to form cyanohydrin complex while NAC is a free radical scavenger and enhances the cellular GSH levels. The study reveals cytotoxicity of cyanide in cells of renal origin and the protective efficacy of A-KG and NAC. PMID- 19136050 TI - Evaluation of alkyloxycarbonyloxymethyl (AOCOM) ethers as novel prodrugs of phenols for topical delivery: AOCOM prodrugs of acetaminophen. AB - The maximum fluxes of a series of alkyloxycarbonyloxymethyl (AOCOM) ethers of acetaminophen (APAP) through hairless mouse skin from isopropyl myristate, IPM (J(MMIPM)) were measured. The J(MMIPM), solubilities in IPM (S(IPM)), water (S(AQ)) and pH 4.0 buffer (S4.0) and molecular weights MW were then fitted to the Roberts-Sloan (RS) equation: log JM = x + y log S(LIPID) + (1-y) log S(AQ)-zMW. Only one of the prodrugs gave an improvement in the flux obtained by APAP itself. The general lack of improvement in flux seems to be due to the fact that there was no improvement in the S(AQ) values of the AOCOM derivatives compared to APAP. When the n = 5 members of the AOCOM series were added to the n = 66 database of J(MMIPM) to give n = 71 and fitted to the RS equation where S(LIPID) was S(IPM), the following coefficients were obtained: x = -0.562, y = 0.501, z = 0.00248, r2 = 0.923. These results demonstrate the importance of improving S(AQ) for prodrugs to improve their solubilities in the skin and hence the flux of the parent drug. The RS equation, which is derived directly from Fick's law, explains this dependence of flux on S(AQ). PMID- 19136049 TI - Investigation of the immunogenicity of p-phenylenediamine and Bandrowski's base in the mouse. AB - p-Phenylenediamine (PPD) exposure is associated with T-cell mediated contact dermatitis. T-cells from allergic patients proliferate following exposure to PPD and the oxido-conjugation product Bandrowski's base (BB). Both compounds are classified as sensitizers in the local lymph node assay; however, because of their instability the nature of the antigenic determinant remains ill-defined. The aim of this study was to explore the immunogenic potential of PPD and BB in mice. Spleen cell proliferation and cytokine secretion was measured ex vivo following antigen recall with soluble PPD or BB and either irradiated or glutaraldehyde fixed, antigen pulsed dendritic cells from syngeneic mice. Glutathione was added to certain incubations. LC-MS analysis and solvent extraction were used to monitor the fate of [(14)C]BB in culture and the extent of BB binding, respectively. Spleen cells from BB exposed, but not PPD- or vehicle-exposed, mice proliferated when stimulated with BB. Proliferating cells secreted high levels of IFN-gamma, GM-CSF and IL-2. Stimulation with PPD instigated low levels of proliferation. Irradiated, but not fixed, dendritic cells pulsed with BB stimulated proliferation signifying a classical hapten mechanism involving irreversible BB binding to protein and processing. BB bound preferentially to serum protein when incubated together with cells and serum. Degradation of BB in the presence of glutathione was associated with a stronger stimulation of specific T-cells at higher BB concentrations. These data demonstrate that BB is a potent immunogen in the mouse. PMID- 19136051 TI - Conformational and bioactivity analysis of insulin: freeze-drying TBA/water co solvent system in the presence of surfactant and sugar. AB - Despite the extensive research into the freeze-drying of aqueous solutions of proteins, it remains unknown whether proteins can survive the lyophilization process in a water-organic co-solvent system and how the process and additives affect the structural stability and activity of the proteins. In the present study, a conformational analysis of insulin in the absence/presence of bile salt and trehalose was carried out, before and after freeze-drying of a tert-butyl alcohol (TBA)/water co-solvent system at volume ratios of TBA to water ranging from 50/50 to 0/100. The study involved the use of ultraviolet derivative and fluorescence spectroscopy, circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. Also the bioactivity of insulin was evaluated in vivo using the streptozotocin (STZ)-induced diabetic mice as an animal model. Initial investigations indicate that the extent of the structural change of insulin depends significantly both on the TBA content and on the concentration of additives, such as sodium deoxycholate, prior to lyophilization. This could be accounted for by the phase behavior properties of the TBA/water co-solvent system, surface denaturation together with the selective and/or forced dispersion of insulin during phase separation. Lyophilized insulin in the presence of bile salt and trehalose retained more of its bioactivity and native-like structure in the solid state compared with that in the absence of additives at various TBA/water ratios, although in all cases there was a major and reversible rearrangement of secondary structure after rehydration, except for insulin at 50% TBA (v/v). Furthermore, both lyophilization in non-eutectic systems and less structural changes in the formulation process lead to more bioactivity. PMID- 19136052 TI - Does perinatal asphyxia induce apoptosis in the inner ear? AB - Pre- and perinatal asphyxia is known to be an important risk factor in the development of neonatal hearing impairment. This study aims to evaluate the role of apoptosis, which is known to play an essential role in the development of the inner ear structures, in the development of neonatal hearing loss caused by pre- and perinatal asphyxia. Eight temporal bones of six different newborns were included. We performed a morphologic analysis by both light microscopy, and transmission electron microscopy, as well as immunohistochemical staining to detect the cleaved form of caspase 3 as apoptosis marker and Bcl 2 as anti apoptotic marker. Early and late phases of apoptosis were evidenced by condensation of chromatin (electron-dense, black structure along nuclear membrane) and fragmentation of the nucleus, respectively. Changes in nuclear morphology during apoptosis correlate with cleavage by caspase 3 located downstream of Bcl 2 action. The immunohistochemistry for cleaved caspase 3 showed a particular predilection for the inner and outer hair cells, spiral ganglion cells and the marginal cells of the stria vascularis. The brain of all examined cases did not show signs of apoptosis. In summary, this investigation suggests that apoptosis takes place before brain tissue apoptosis and is probably an earlier event than thought. Apoptosis of the cochlea is known to play an essential role in the development of the inner ear. Additionally, this study shows that apoptosis may play an important role in the development of hearing impairment, caused by pre- and perinatal asphyxia. PMID- 19136053 TI - Evidence for differential regulation of lactate metabolic properties in aged and unloaded rat skeletal muscle. AB - Skeletal muscles of elderly individuals show fatigue resistance and reduced lactate accumulation compared with those of young subjects during activities that recruit a small amount of muscle mass. To explore the mechanism underlying the functional changes in aged muscle, we focused on lactate metabolic properties, including monocarboxylate transporter (MCT) 1 and MCT4, in muscles from old and young control rats and hindlimb-suspended young rats. MCT1 expression was lower in soleus (SOL) of old rats than in SOL of young control rats, but was similar in young control and hindlimb-suspended rats. MCT4 expression was lower in extensor digitorum longus (EDL) of old rats than in that of young control rats, but did not differ between young control and hindlimb-suspended rats. The ratio of lactate dehydrogenase to citrate synthase activities was higher in SOL of hindlimb-suspended and old rats than in SOL of young control rats, and was lower in EDL of old rats than in those of young control and hindlimb-suspended rats. Our data suggest that aging causes metabolic changes that can reduce lactate accumulation during exercise and increase fatigue resistance in skeletal muscle, and that these changes result from aging rather than from inactivity. PMID- 19136054 TI - Effects of (+)-methamphetamine on path integration and spatial learning, but not locomotor activity or acoustic startle, align with the stress hyporesponsive period in rats. AB - Rats treated with (+)-methamphetamine (MA) on postnatal days (P) 11-20 exhibit long-term spatial and path integration (Morris water maze (MWM) and Cincinnati water maze (CWM)) learning deficits whereas those treated on P1-10 do not. MA treatment increases corticosterone release in an age-dependent U-shaped pattern that corresponds to the stress hyporesponsive period (SHRP; P4-15). Here we tested the hypothesis that the cognitive effects induced by MA are associated with treatment that begins within the SHRP. Three treatment regimens were compared, P1-10, P6-15, and P11-20. One male/female pair/litter received 0, 10, or 25mg/kg MA/dose (four doses/day at 2h intervals given s.c. with 19-21 litters/regimen). Locomotor activity and acoustic startle were tested as behaviors not predicted to be associated with the SHRP. Cincinnati and Morris water maze findings were consistent with the hypothesis in that MA-treated animals exposed from P6-15 or P11-20 showed impaired learning compared to those exposed from P1-10; however, on probe trials in the Morris water maze, MA-induced memory impairments were not regimen-specific and were contributed to by all treatment regimens. All MA treatment regimens induced reductions in locomotor activity and acoustic startle facilitation as expected. No differential effect on prepulse trials was seen suggesting no impairment in sensory gating. Cognitive deficits from neonatal MA treatment are associated with the SHRP and may be the product of hypothalamic-pituitary-adrenal (HPA) axis dysregulation during critical periods of brain development. PMID- 19136056 TI - Development of a species-specific colorimetric-PCR assay for detection and species differentiation of Mycobacterium immunogenum and Mycobacterium chelonae and its comparison with quantitative real-time PCR for field metalworking fluids. AB - Mycobacterium immunogenum and Mycobacterium chelonae are closely related species associated with occupational hypersensitivity pneumonitis (HP) and nosocomial infections. There is a need to develop specific and readily adaptable methods for detection and speciation of these agents. Here we report development of a probe based colorimetric-PCR assay involving heat shock protein (hsp) gene amplification (228 bp) and its detection in an ELISA-like reaction. A quantitative format of this assay was developed and validated on metalworking fluids (MWF). The assay showed a minimum detection limit of 10 fg genomic DNA or 1 mycobacterial cell, albeit with variations depending on type and composition of the MWF matrix. When applied to the field MWF samples, the developed assay was found to be comparable to the real-time PCR assay, and allowed direct speciation of MWF mycobacteria without sequencing and/or restriction pattern analysis. In conclusion, the developed colorimetric PCR allows detection and quantification of MWF mycobacteria without culturing and is the first probe-based assay for unambiguous differentiation between the two phylogenetically closely related species, M. immunogenum and M. chelonae. Considering that the assay offers high throughput format involving relatively simpler instrument infrastructure, it has a potential for applications in routine assessment of MWF mycobacteria in diagnostic and industrial laboratories. PMID- 19136055 TI - Increased IgG4 levels in children with autism disorder. AB - Accumulating evidence indicates that immune dysfunction is associated with autism disorders in a significant subset of children. Previous reports have shown abnormal immunoglobulin (Ig) levels, including an increased presence of autoreactive antibodies in the circulation of individuals with autism. As IgG is the predominant antibody isotype in circulation, we expected that an altered immune response could result in an abnormal IgG subclass profile in children with autism. We examined circulating plasma levels of IgG1, IgG2, IgG3, and IgG4 in 241 children from the CHARGE (Childhood Autism Risks from Genetics and the Environment) study, a large epidemiologic case-control investigation, including 114 children who meet full criteria for autism disorder (AU), 96 typically developing control children (TD) from a randomly selected sample of the general population, and 31 children with developmental delays (DD). We report significantly increased levels of the IgG4 subclass in children with AU compared with TD control children (p=0.016) and compared with DD controls (p=0.041). These results may suggest an underlying immunological abnormality in AU subjects resulting in elevated IgG4 production. Further investigation is necessary to elucidate the relationship between immunological findings and behavioral impairments in autism. PMID- 19136057 TI - Erythropoietin protects retinal pigment epithelial cells from oxidative damage. AB - Oxidative damage from reactive oxygen species (ROS) has been implicated in many diseases, including age-related macular degeneration, in which the retinal pigment epithelium (RPE) is considered a primary target. The aim of this study was to determine whether erythropoietin (EPO) protects cultured human RPE cells against oxidative damage and to identify the pathways that may mediate protection. EPO (1 IU/ml) significantly increased the viability of oxidant treated RPE cells, decreased the release of the inflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta, recovered the RPE cells' barrier integrity disrupted by oxidative stress, prevented oxidant-induced cell DNA fragmentation and membrane phosphatidylserine exposure, and also reduced the levels of oxidant-induced intracellular ROS and restored cellular antioxidant potential, total antioxidant capacity, glutathione peroxidase, and superoxide dismutase and decreased malondialdehyde, the end product of lipid peroxidation. EPO inhibited caspase-3-like activity. Protection by EPO was partly dependent on the activation of Akt1 and the maintenance of the mitochondrial membrane potential. No enhanced or synergistic protection was observed during application of Z-DEVD-FMK (caspase-3 inhibitor) combined with EPO compared with cultures exposed to EPO and H(2)O(2) alone. Together, these results suggest that EPO could protect against oxidative injury-induced cell death and mitochondrial dysfunction in RPE cells through modulation of Akt1 phosphorylation, mitochondrial membrane potential, and cysteine protease activity. PMID- 19136058 TI - Tubulin acetylation favors Hsp90 recruitment to microtubules and stimulates the signaling function of the Hsp90 clients Akt/PKB and p53. AB - Involved in a wide range of cellular processes such as signal transduction, microtubules are highly dynamic polymers that accumulate various post translational modifications including polyglutamylation, polyglycylation, carboxyterminal cleavage and acetylation, the functions of which just begin to be uncovered. The molecular chaperone Hsp90, which is essential for the folding and activity of numerous client proteins involved in cell proliferation and apoptosis, associates with the microtubule network but the effects of tubulin post-translational modifications on its microtubule binding has not yet been investigated. Herein, we show that both the constitutive (beta) and the inducible (alpha) Hsp90 isoforms bind to microtubules in a way that depends on the level of tubulin acetylation. Tubulin acetylation also stimulates the binding and the signaling function of at least two of its client proteins, the kinase Akt/PKB and the transcription factor p53. This study highlights the role of tubulin acetylation in modulating microtubule-based transport of Hsp90-chaperoned proteins and thus in regulating signaling dynamics in the cytoplasm. PMID- 19136059 TI - Role of polyamines in p53-dependent apoptosis of intestinal epithelial cells. AB - Although p53 is known to play a critical role in the proliferation of gastrointestinal epithelia, the role of the Mdm2/p53 pathway in response to inducers of apoptosis in intestinal epithelial cells is unknown. Our data show that camptothecin (CPT)-induced apoptosis correlated with increased p53, p21Cip1, and Mdm2 protein levels, with a simultaneous increase in ATR Ser428, p53 Ser15 and Mdm2 Ser166 phosphorylation in IEC-6 cells. Increased p53 levels and its phosphorylation increased Bax protein, caspase-9, -3 activation and apoptosis. However, TNF-alpha/CHX-mediated apoptosis was independent of p53 protein levels and phosphorylation. The translation inhibitor, cycloheximide (CHX), prevented CPT-induced apoptosis. CHX completely prevented CPT-induced p53 phosphorylation and synthesis of p21Cip1, Bax and Bcl-xL proteins without altering p53 levels. The p53 activator, RITA, augmented CPT-induced apoptosis. The Mdm2 antagonist, Nutlin-3, significantly increased apoptosis, which was accompanied by increased p53, Mdm2 and p21Cip1 protein levels. The ATM/ATR kinase inhibitor, CGK733, blocked CPT-induced p53 Ser15 phosphorylation and protected cells from CPT induced apoptosis. Inhibition of ornithine decarboxylase (ODC) with alpha difluromethylornithine (DFMO) and subsequent depletion of intracellular polyamines increased p53 protein, Mdm2 Ser166 phosphorylation and conferred resistance to CPT-induced apoptosis. However, polyamine depletion had no effect on p53 phosphorylation. Nutlin-3 reversed the protective effect of DFMO and sensitized cells to CPT-induced apoptosis. These results suggest that p53 stabilization and accumulation in response to polyamine depletion predominantly modulate cell cycle checkpoints via p21Cip1 expression and inhibit transcription of target genes responsible for apoptosis. In contrast, phosphorylation and stabilization of p53 in response to DNA-damage lead to apoptosis, which indicates different roles of p53 during DNA damage and polyamine depletion. PMID- 19136060 TI - Lysophosphatidic acid induces Ca2+ mobilization and c-Myc expression in mouse embryonic stem cells via the phospholipase C pathway. AB - Embryonic stem cells (ESC) are pluripotent and could be maintained in vitro in a self-renewing state indefinitely, at the same time preserving their potential to differentiate towards more specific lineages. Despite the progress in the field, the complex network of signalling cascades involved in the maintenance of the self-renewing and pluripotent state remains not fully understood. In the present study, we have investigated the role of lysophosphatidic acid (LPA), a potent mitogen present in serum, in Ca(2+) signalling and early gene activation in mouse ESC (mESC). In these cells, we detected the expression of the G-protein coupled LPA receptor subtypes LPA(1), LPA(2) and LPA(3). Using fluorescence Ca(2+) imaging techniques, we showed that LPA induced an increase in intracellular Ca(2+) concentration. This increase was also observed in the absence of extracellular Ca(2+), suggesting the involvement of internal stores. Pre treatment with BAPTA-AM, thapsigargin or U-73122 efficiently blocked this Ca(2+) release, indicating that LPA was evoking Ca(2+) mobilization from the endoplasmic reticulum via the phospholipase C (PLC) pathway. Interestingly, this signalling cascade initiated by LPA was involved in inducing the expression of the Ca(2+) dependent early response gene c-myc, a key gene implicated in ESC self-renewal and pluripotency. Additionally, LPA increased the proliferation rate of mESC. Our findings therefore outline the physiological role of LPA in mESC. PMID- 19136061 TI - Antibiotic delivery polyurethanes containing albumin and polyallylamine nanoparticles. AB - Nano-structured polymers delivering an antibiotic for the prevention of medical device-related infections were developed. Systems consisted of bovine serum albumin or polyallylamine nanoparticles alone or entrapped in a polyurethane and then loaded with cefamandole nafate, chosen as a drug model. Results showed that nanoparticles alone were able to adsorb high antibiotic amounts due to their high surface/volume ratio. However, they released cefamandole in an uncontrolled fashion, leading to a rapid loss of antibacterial activity. Improvements in the release control were obtained when CEF loaded and non-loaded nanoparticles were entrapped in a carboxylated polyurethane. For these systems the drug delivery was at least of 50% with respect to nanoparticles alone with a prolonged antimicrobial activity up to 9 days. PMID- 19136063 TI - Gene cloning, expression and homology modeling of hemolysin gene from Aeromonas hydrophila. AB - Hemolysin is a significant toxin secreted by Aeromonas hydrophila, which contributes pathogenicity of fish to humans. The complete ORF of hemolysin gene (1886 bp) was amplified using PCR. It was cloned in TA and sub-cloned in pET28a vector then transformed into Escherichia coli BL21(DE3) codon plus RP cells expressed by the induction with 1.0 mM of IPTG. The expected size of expressed protein was 68.0 kDa estimated by migration in 12% SDS-PAGE. Anti-His monoclonal antibodies were used to substantiate the recombinant protein by Western blotting. The percent similarity between hemolysin of A. hydrophila with other hemolytic toxins revealed that the hemolysin/aerolysin/cytotoxin sequence varied from 99.35 to 50.40%. Homology modeling was used to construct 3-D structure of hemolysin of A. hydrophila with the known crystal 3-D structure (PDB: 1XEZ). This protein can be used for immunoassays and it is suitable for vaccine candidate against A. hydrophila infection. PMID- 19136062 TI - Gamma-protocadherin homophilic interaction and intracellular trafficking is controlled by the cytoplasmic domain in neurons. AB - Gamma-protocadherins (Pcdh-gammas) are good candidates to mediate specificity in synaptogenesis but their role in cell-cell interactions is a matter of debate. We proposed that Pcdh-gammas modify preformed synapses via trafficking of Pcdh gammas-containing organelles, insertion into synaptic membranes and homophilic transcellular interaction. Here we provide evidence in support of this model. We show for the first time that Pcdh-gammas have homophilic properties and that they accumulate at dendro-dendritic and axo-dendritic interfaces during neuronal development. Pcdh-gammas are maintained in a substantial mobile intracellular pool in dendrites and cytoplasmic deletion shifts the molecule to the surface and reduces the number and velocity of the mobile packets. We monitored Pcdh-gamma temporal and spatial dynamics in transport organelles. Pcdh-gamma organelles bud and fuse with stationary clusters near synapses. These results suggest that Pcdh gamma-mediated cell-cell interactions in synapse development or maintenance are tightly regulated by control of intracellular trafficking via the cytoplasmic domain. PMID- 19136064 TI - [18F]flumazenil binding to central benzodiazepine receptor studies by PET- quantitative analysis and comparisons with [11C]flumazenil. AB - [(11)C]flumazenil is the reference radioligand for Positron Emission Tomography (PET) studies of central benzodiazepine (BZ) receptors. Fluorine is available in the flumazenil molecule and [(18)F]flumazenil has recently been prepared. The aim of the present PET-study in 8 male subjects was to examine the binding of [(18)F]flumazenil in the human brain by direct comparison with [(11)C]flumazenil. Each subject participated in two 93-minute PET-measurements with [(11)C]flumazenil and [(18)F]flumazenil, respectively. Data were analyzed using compartment models with metabolite-corrected arterial plasma input and reference tissue models using the pons as reference region. There was no evident difference between the kinetic behaviors of the two ligands. Overall, the noise in the time activity curves for [(18)F]flumazenil was lower at late time points, and the variance of the kinetic parameters was lower than for [(11)C]flumazenil. In BZ receptor rich regions, such as the neocortex, the 3-compartment model was statistically favored, whereas the 2-compartment model was favored in the pons. Binding potential values obtained by the reference tissue models were in good agreement with those obtained by the kinetic analysis. There was no support for the presence of specific binding in the pons. In conclusion, the binding and the kinetic behavior of [(11)C]flumazenil and [(18)F]flumazenil were similar. The present analysis supports the use of pons as reference region in simplified protocols without arterial blood sampling. [(18)F]flumazenil should thus be an excellent choice for applied studies at centers not having a cyclotron. PMID- 19136065 TI - Phylogenetic relationships of Sardinian cave salamanders, genus Hydromantes, based on mitochondrial and nuclear DNA sequence data. PMID- 19136066 TI - Patterns of plant speciation in the Cape floristic region. AB - Plant species have accumulated in the Cape region of southern Africa to a much greater degree than in areas of equivalent size in the rest of the subcontinent. Although this could be a consequence simply of lower extinction rates in the Cape, most researchers have invoked high rates of ecological speciation, driven by unique aspects of the Cape environment, as the primary explanation for this richness. To assess these ideas, we analyzed the frequencies of ecological shifts among 188 sister species pairs obtained from molecular phylogenies of eight Cape clades. Ecological shifts were evident in 80% of sister species pairs, with general habitat, pollinator, and fire-survival strategy shifts being especially frequent. Contrary to an established idea that shifts in soil type are frequently associated with speciation of Cape taxa, these shifts were relatively rare, occurring in just 17% of species pairs. More cases of sister species divergence are accompanied solely by floral than by vegetative diversification, suggesting an important role for pollinator-driven speciation. In an analysis of two large orchid genera that have radiated in both the Cape and the rest of southern Africa, the frequency of ecological shifts (general habitat, soil type, altitude and flowering time), did not differ between sister species pairs in the Cape region and those outside it. Despite suggestions that Cape plants tend to have small range sizes and show fine-scale patterns of speciation, range size did not differ significantly between species in the Cape and those outside it. We conclude that ecological speciation is likely to have been important for radiation of the Cape flora, but there is no evidence as yet for special "Cape" patterns of ecological speciation. PMID- 19136067 TI - Mitochondria in early mammalian development. AB - The role of mitochondria as central determinants of development competence of oocytes and preimplantation stage embryos is considered in the context of the diverse activities these organelles have in normal cell function. Stage- and cell cycle-specific mitochondrial translocations and redistributions are described with respect to mechanisms of cytoplasmic remodeling that may establish domains of autonomous regulation of mitochondrial function and activity during early development. The functions of mitochondria as intracellular signaling elements, as regulators of signaling pathways, and oxygen sensors in differentiated cells are suggested to have similar capacities during mammalian oogenesis and early embryogenesis. Questions concerning the numerical size of the oocyte mitochondrial complement, the energy required to support normal preovulatory oogenesis and preimplantation embryogenesis, and the regulation of mitochondrial activity by extrinsic and intrinsic factors are addressed with respect the potential they may have for new investigational approaches to study the origin of the differential developmental competence of human oocytes and preimplantation stage embryos. PMID- 19136068 TI - Mitochondria and the redox control of development in cnidarians. AB - Mitochondria are the product of an ancient symbiosis between bacteria and host cells. While mitochondria function primarily in energy conversion, increasing amounts of evidence indicate that mitochondrial metabolic state can influence various emergent features of eukaryotic cells. Important intermediaries in such redox signaling include by-products of metabolism, particularly reactive oxygen species (ROS). This review uses cnidarians, a group of basally branching animals, to illustrate the many and varied effects of ROS on development. ROS from both mitochondria and algal symbionts are considered. Because some applications of ROS may lack specificity, the signaling demands of mitochondria and algae may to some extent conflict. An extensive algal symbiosis may thus be incompatible with a well-developed capacity for mitochondrial signaling. PMID- 19136069 TI - Semantics and types of cough. AB - The panel considered the different types of cough in terms of basic mechanisms and clinical manifestations; both experimentally and clinically cough could occur in single efforts and as 'bouts' or 'epochs'. There were different definitions of cough but, provided the definition used was clear, this did not seem to be a major concern. The methods available for determining the nature or type of clinical cough were discussed, in particular automated cough counting in the clinic and more sophisticated methods available in the laboratory. With regard to semantics, there has been great variation in the names used; this applies to nervous sensors for cough, to cough reflexes and epochs, to clinical names for cough, and to cough sounds. Some simplification and uniformity of nomenclature seemed desirable although, provided the use of a name was clear, little confusion probably existed. The panel felt that the cough nomenclature would evolve with time and would prove to be useful for investigators, clinicians and coughers. PMID- 19136070 TI - Airway smooth muscle contraction - perspectives on past, present and future. AB - Past and contemporary views of airway smooth muscle (ASM) have led to a high level of understanding of the control and intracellular regulation of force or shortening of ASM and of its possible role in airway disease. As well as the multitude of cellular mechanisms that regulate ASM contraction, a number of structural and mechanical factors, which are only present at the airway and lung level, provide overriding control over ASM. With new knowledge about the cellular physiology and biology of ASM, there is increasing need to understand how ASM contraction is regulated and expressed at these airway and system levels. PMID- 19136071 TI - Schistosoma mansoni infection alters co-stimulatory molecule expression and cell activation in asthma. AB - Chronic schistosomiasis induces Th2/T regulatory responses which are able to down modulate allergic inflammation and asthma. Because co-stimulatory molecules and IL-10 are essential for inducing tolerance, the aim of this study was to determine by flow cytometry, the expression of CD28, CTLA4, CD40L, CD80, CD86, HLA-DR, IL-10 and IL-10 receptor, by mononuclear cells from asthmatic individuals infected with Schistosoma mansoni and compare with non-infected individuals. Peripheral blood mononuclear cells were stained with fluorochrome conjugated antibodies for the expression of co-stimulatory molecules, and for intracellular CTLA4 and IL-10 expression. There was no significant difference in the frequency of T cells expressing CD28 between the two groups. However, the frequency of TCD4(+) cells expressing CTLA4 and CD40L was higher in infected asthmatics. The frequency of monocytes expressing CD80 and CD86 did not differ between groups, while the expression of HLA-DR and IL-10 receptor was higher on monocytes of infected individuals. Furthermore, monocytes and CD4(+)CD25(+) cells of infected individuals expressed higher levels of IL-10. We conclude that, besides alternatively-activated monocytes that are, together with CD4(+)CD25(+) cells, important sources of IL-10, CTLA4 and CD40L expression may also participate in the down-modulation of inflammatory allergic response in S. mansoni-infected asthmatics. PMID- 19136073 TI - Differential sub-nuclear localisation of repressive and activating histone methyl modifications in P. falciparum. AB - Post-translational histone modifications and sub-nuclear organization epigenetically influence gene regulation, especially those implicated in antigenic variation of Plasmodium falciparum. Here we screened for histone methylation modifications and determined, for the first time, their spatial nuclear localisation. Differential enrichment in sub-nuclear compartments, suggesting high local concentrations of particular methyl marks was observed. H3 K79me3 particularly, does not co-localise with already defined nuclear compartments, thus apparently defining a compartment not observed previously in other eukaryotes. Our data show the presence of discrete sub-nuclear zones enriched for histone methyl marks, pointing to the existence of specialized transcription factories and repressive regions in Plasmodium. PMID- 19136072 TI - Phenotypic studies on recombinant human immunodeficiency virus type 1 (HIV-1) containing CRF01_AE env gene derived from HIV-1-infected patient, residing in central Thailand. AB - Human immunodeficiency virus type 1 (HIV-1) env genes were cloned from blood samples of HIV-1-infected Thai patients, and 35 infectious CRF01_AE envelope glycoprotein (Env)-recombinant viruses were established. In this report, we examined the neutralization susceptibility of these viruses to human monoclonal antibodies, 2G12, IgG1 b12, 2F5 and 4E10, pooled patient plasma, coreceptor antagonists and fusion inhibitor, T-20. The neutralization susceptibility of CRF01_AE Env-recombinant viruses to 2F5, 4E10, patient plasma, coreceptor antagonists and T-20 varied, while most viruses showed low susceptibility to 2G12 and IgG1 b12. Several dual-tropic viruses showed lower susceptibility to 2F5 and 4E10 than CXCR4- or CCR5-tropic viruses. Neutralization susceptibility of the CRF01_AE Env-recombinant virus to pooled patient plasma was negatively correlated with the length of the V1/V2 region or the number of potential N-linked glycosylation sites in conserved regions of gp120. No correlation was found between the coreceptor usage and neutralization susceptibility of the virus to T 20, whereas several dual-tropic viruses showed higher susceptibility to coreceptor antagonists than CXCR4- or CCR5-tropic viruses. We propose that these CRF01_AE Env-recombinant viruses are useful to further study the molecular mechanism of the susceptibility of CRF01_AE Env to neutralizing antibodies and viral entry inhibitors. PMID- 19136074 TI - Connexin 43 a potential regulator of cell proliferation and apoptosis within the seminiferous epithelium. AB - The gap junction proteins, connexins (Cx), are present in the testis and among them Cx43 play an essential role in spermatogenesis. By using an in vitro proliferation model of germ cells and Sertoli cells, we tempted here to clarify the role of Cx43 in the control of Sertoli and germ cell proliferation and apoptosis. Cx43 was detected in purified preparations of Sertoli cells and spermatogonia and immunolocalized in both cell types identified by vimentin and c kit, respectively. Inhibition of gap junction coupling by the gap junction inhibitor alpha-GA significantly enhanced BrdU incorporation in Sertoli cells and reduced the number of activated caspase-3 positive germ cells. Similarly, inhibitory Cx43 and pan-Cx mimetic inhibitory peptides increased proliferation of Sertoli cells and stimulated survival of germ cells. Cx32 mimetic inhibitory peptide also stimulated Sertoli cell proliferation without altering germ cell proliferation and apoptosis. The present results reveal that Cx43 gap junctions between Sertoli cells participate in the control of Sertoli cell proliferation and that Cx43 gap junctions between Sertoli cells and spermatogonia are indirectly involved in germ cell number increase by controlling germ cell survival rather than germ cell proliferation. PMID- 19136075 TI - Regulation of hepatic fatty acid elongase 5 by LXRalpha-SREBP-1c. AB - Dietary essential fatty acids linoleic acid and alpha-linolenic acid are converted to arachidonic-, eicosapentaenoic-, and docosahexaenoic acid under tight regulation by nutritional status and hormones. Hepatic fatty acid elongase 5 (Elovl5) elongates C18-20 polyunsaturated fatty acids (PUFAs) and is important for biosynthesis of C20-22 PUFAs. We demonstrate that Liver X Receptor alpha (LXRalpha) and sterol regulatory binding protein-1c (SREBP-1c) regulate hepatic Elovl5 expression. LXRalpha and LXRbeta play different roles in maintenance of basal expression of Elovl5. LXRalpha is necessary for basal as well as LXR agonist induced Elovl5 transcription. Promoter studies revealed that the mouse Elovl5 gene is a direct SREBP-1c target. The up-regulation of Elovl5 expression by LXR agonist is likely secondary to the induction of SREBP-1c. PUFAs repress expression of SREBP-1c and Elovl5, but when combined with LXR ligand stimulation, which increases SREBP-1c mRNA and nuclear SREBP-1c, Elovl5 mRNA levels are restored to normal. Our studies suggest that an LXRalpha-SREBP-1c pathway plays a regulatory role in hepatic biosynthesis of PUFAs through transcriptional activation of Elovl5 as well as other desaturases. The stimulatory role of LXRalpha-SREBP-1c in the production of PUFAs enables the possibility for a feedback regulation of hepatic lipogenesis through PUFA mediated repression of SREBP-1c expression. PMID- 19136076 TI - Transformation of 5-ene steroids by the fungus Aspergillus tamarii KITA: mixed molecular fate in lactonization and hydroxylation pathways with identification of a putative 3beta-hydroxy-steroid dehydrogenase/Delta5-Delta4 isomerase pathway. AB - The fungus Aspergillus tamarii metabolizes progesterone to testololactone in high yield through a sequential four step enzymatic pathway which, has demonstrated flexibility in handling a range of steroidal probes. These substrates have revealed that subtle changes in the molecular structure of the steroid lead to significant changes in route of metabolism. It was therefore of interest to determine the metabolism of a range of 5-ene containing steroidal substrates. Remarkably the primary route of 5-ene steroid metabolism involved a 3beta-hydroxy steroid dehydrogenase/Delta(5)-Delta(4) isomerase (3beta-HSD/isomerase) enzyme(s), generating 3-one-4-ene functionality and identified for the first time in a fungus with the ability to handle both dehydroepiansdrosterone (DHEA) as well as C-17 side-chain containing compounds such as pregnenolone and 3beta hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one. Uniquely in all the steroids tested, 3beta-HSD/isomerase activity only occurred following lactonization of the steroidal ring-D. Presence of C-7 allylic hydroxylation, in either epimeric form, inhibited 3beta-HSD/isomerase activity and of the substrates tested, was only observed with DHEA and its 13alpha-methyl analogue. In contrast to previous studies of fungi with 3beta-HSD/isomerase activity DHEA could also enter a minor hydroxylation pathway. Pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn 5-en-20-one were metabolized solely through the putative 3beta-HSD/isomerase pathway, indicating that a 17beta-methyl ketone functionality inhibits allylic oxidation at C-7. The presence of the 3beta-HSD/isomerase in A. tamarii and the transformation results obtained in this study highlight an important potential role that fungi may have in the generation of environmental androgens. PMID- 19136077 TI - Catalytic properties of the expressed acyclic carotenoid 2-ketolases from Rhodobacter capsulatus and Rubrivivax gelatinosus. AB - Purple photosynthetic bacteria synthesize the acyclic carotenoids spheroidene and spirilloxanthin which are ketolated to spheroidenone and 2,2' diketospirilloxanthin under aerobic growth. For the studies of the catalytic reaction of the ketolating enzyme, the crtA genes from Rubrivivax gelatinosus and Rhodobacter capsulatus encoding acyclic carotenoid 2-ketolases were expressed in Escherichia coli to functional enzymes. With the purified enzyme from the latter, the requirement of molecular oxygen and reduced ferredoxin for the catalytic activity was determined. Furthermore, the putative intermediate 2-HO-spheroidene was in vitro converted to the corresponding 2-keto product. Therefore, a monooxygenase mechanism involving two consecutive hydroxylation steps at C-2 were proposed for this enzyme. By functional pathway complementation studies in E. coli and enzyme kinetic studies, the product specificity of both enzymes were investigated. It appears that the ketolases could catalyze most intermediates and products of the spheroidene and spirilloxanthin pathway. This was also the case for the enzyme from Rba. capsulatus from which spirilloxanthin synthesis is absent. In general, the ketolase of Rvi. gelatinosus had a better specificity for spheroidene, HO-spheroidene and spirilloxanthin as substrates than the ketolase from Rba. capsulatus. PMID- 19136078 TI - Transplantation tolerance induced by regulatory T cells: in vivo mechanisms and sites of action. AB - The mechanisms by which CD4(+)CD25(+)Foxp3(+) T (Treg) cells regulate effector T cells in a transplantation setting and their in vivo homeostasis still remain to be clarified. Using a mouse adoptive transfer model, we analyzed the in vivo expansion, trafficking, and effector function of alloreactive T cells and donor specific Treg cells, in response to a full-thickness skin allograft. Fluorescent labeled CD4(+)CD25(-) and antigen-specific Treg cells were transferred alone or co-injected into syngeneic BALB/c-Nude recipients transplanted with skins from (C57BL/6 x BALB/c) F1 donors. Treg cells divided in vivo, migrated and accumulated in the allograft draining lymph nodes as well as within the graft. The co-transfer of Treg cells did not modify the early activation and homing of CD4(+)CD25(-) T cells in secondary lymphoid organs. However, in the presence of Treg cells, alloreactive CD4(+)CD25(-) T cells produced significantly less IFN gamma and were present in reduced numbers in the secondary lymphoid organs. Furthermore, time-course studies showed that Treg cells were recruited into the allograft at a very early stage after transplantation and effectively prevented the infiltration of effector T cells. In conclusion, suppression of rejection requires the early recruitment to the site of antigenic challenge of donor specific Treg cells, which then mainly regulate the effector arm of T cell alloresponses. PMID- 19136079 TI - Lung compression effects on gas exchange in human breath-hold diving. AB - Lung compression during breath-hold diving reduces gas exchanging surface area. Beyond a critical depth, collapse of all alveoli should result in total pulmonary shunt and a drop in arterial oxygen partial pressure toward the mixed-venous level. The effect of lung collapse on human breath-hold diving capability is analysed using a computational model of the lungs and circulation that simulates oxygen, carbon dioxide, and nitrogen exchange between alveoli, blood, and tissues. Gas uptake during descent becomes limited by lung compression when the ratio of diffusing capacity to the product of perfusion and gas solubility in blood drops below one. An equation is derived for estimating collapse depth due to direct alveolar compression and time-dependent absorption atelectasis. Oxygen dissolved in blood during descent builds a limited capacitive store for supporting metabolism during the period of lung collapse. Hypoxemia with loss of consciousness prior to alveolar re-opening on ascent is predicted to occur on dives beyond 300 m, depending on initial lung volume. PMID- 19136080 TI - Comparisons of effects of intravenous and inhaled methacholine on airway physiology in a murine asthma model. AB - Airway responses to intravenous (i.v.) and inhaled (i.h.) delivery of methacholine (MCh) in BALB/c and C57BL/6 mouse strains have been compared with and without ovalbumin (OVA)-induced airway inflammation. Bronchial reactivity to MCh was assessed in anaesthetised and tracheostomised animals by using an animal ventilator (flexiVent). We partitioned the response of the lungs into airway and parenchymal components in order to compare the contributions of the airways with those of the lung parenchyma to the pulmonary mechanical responses resulting from different routes of MCh administration. Our results indicate disparate physiological responses. Intravenous MCh delivery induced a higher maximum lung resistance than i.h. MCh in OVA-treated BALB/c mice but not in C57BL/6 mice. Inhaled MCh delivery led to a significantly larger fall in lung compliance and a greater impact on peripheral airways than i.v. MCh in both strains. In conclusion, i.v. and i.h. MCh produced disparate effects in different murine strains and variant responses in inflamed airways and healthy controls. The two methods of MCh delivery have important advantages but also certain limitations with regard to measuring airway reactivity in a murine model of allergic asthma. PMID- 19136081 TI - Proteome analysis of a human liver carcinoma cell line stably expressing hepatitis delta virus ribonucleoproteins. AB - Hepatitis delta virus (HDV) infects human hepatocytes already infected with the hepatitis B virus increasing about ten fold the risk of cirrhosis and fulminant hepatitis. The lack of an appropriate cell culture system capable of supporting virus replication has so far impaired the detailed investigation of the HDV biology including the identification of host factors involved in pathogenesis. Here, we made use of a HDV cDNA stably transfected cell line, Huh7-D12, in a proteomic approach to identify the changes in the protein expression profiles in human liver cells that arise as a consequence of HDV replication. Total protein extracts from Huh7-D12 cells and of the corresponding non transfected human liver carcinoma cell line, Huh7, were separated by 2-DE. Differentially expressed spots were identified by MALDI-TOF followed by database searching. We identified 23 differentially expressed proteins of which 15 were down regulated and 8 up regulated in Huh7-D12 cells. These proteins were found to be involved in different cellular pathways. The down regulation of the histone H1-binding protein and of triosephosphate isomerase was confirmed by Real time PCR, and the up regulation of the La protein and lamin A/C was validated by western blot. PMID- 19136082 TI - Cortical bone development in black and white South African children: iliac crest histomorphometry. AB - Fragility fracture rates in South Africa are lower in blacks (B) than in whites (W) both in adults and in children. In adults this difference may in part be explained by histomorphometric findings in iliac crest cortical bone of B of thicker, less porous cortices, greater endocortical (Ec) wall thickness, fewer canals and greater osteoid thickness accompanied by greater mineral apposition rate and bone formation rate compared to W. Since no comparative data for B and W children are available we examined iliac crest cortical bone of 57 B and 56 W aged 0-23 yrs by routine histomorphometry. RESULTS: The effects of growth as expressed in differences between external and internal cortex were similar in B and W children. Cortical thickness increased with age similarly in B and W until about age 15 whereafter it continued to increase only in B. Ec wall thickness rose with age in B but did not change in W. After age 11 canal number was lower in B. Cortical porosity was highest between ages 6 and 15 with a tendency to lower values in the external cortex in B. Thus structural differences reported in adults were evident in children. Bone turnover as reflected in osteoid surface and eroded surface declined with age similarly in B and W but osteoid thickness did not change with age. Greater osteoid thickness in B children could reflect greater vigor of osteoblasts and greater osteoblast team performance as it did in B adults and may have contributed to the structural advantage in B children. CONCLUSION: B children showed greater values for osteoid thickness, endocortical wall thickness and cortical thickness, and a tendency to lower porosity compared to W children. These features may contribute to lower fragility fracture rates in B children. Differing environmental influences and possibly genetic effects may play a role. PMID- 19136083 TI - Wnt signaling and osteoarthritis. AB - Osteoarthritis is a common disease, clinically manifested by joint pain, swelling and progressive loss of function. The severity of disease manifestations can vary but most of the patients only need intermittent symptom relief without major interventions. However, there is a group of patients that shows fast progression of the disease process leading to disability and ultimately joint replacement. Apart from symptom relief, no treatments have been identified that arrest or reverse the disease process. Therefore, there has been increasing attention devoted to the understanding of the mechanisms that are driving the disease process. Among these mechanisms, the biology of the cartilage-subchondral bone unit has been highlighted as key in osteoarthritis, and pathways that involve both cartilage and bone formation and turnover have become prime targets for modulation, and thus therapeutic intervention. Studies in developmental, genetic and joint disease models indicate that Wnt signaling is critically involved in these processes. Consequently, targeting Wnt signaling in a selective and tissue specific manner is an exciting opportunity for the development of disease modifying drugs for osteoarthritis. PMID- 19136084 TI - Rectal misoprostol vs 15-methyl prostaglandin F2alpha for retained placenta after second-trimester delivery. AB - OBJECTIVE: The purpose of this study was to compare rectal misoprostol (RM) with 15-methyl prostaglandin F2alpha (PGF(2alpha)) for the management of retained placenta after second-trimester deliveries. STUDY DESIGN: A retrospective study of all second-trimester deliveries between the years 2000 and 2005 was performed. Women were divided into 2 groups, depending on whether they received RM or PGF(2alpha) after the delivery. RESULTS: Three hundred three second-trimester deliveries were analyzed. The time from the administration of medications to the placental delivery was significantly shorter in women who received PGF(2alpha), compared with the RM group (49.5 vs 89 minutes; P < .01). Women who received PGF(2alpha) had lower rates of retained placenta (4.9% vs 12.4%; P = .02). CONCLUSION: The use of PGF(2alpha) after second-trimester deliveries results in shorter third stage of labor and lower rates of retained placenta compared with RM. PMID- 19136085 TI - A role for menstruation in preconditioning the uterus for successful pregnancy. AB - Menstruation is widely viewed as serving no purpose other than to reinitiate the endometrial cycle in the absence of pregnancy. Yet, it is striking that cyclic endometrial decidualization followed by menstrual shedding is confined to the few species, including human beings, where placenta formation entails deep trophoblast invasion of maternal tissues and its vasculature. Both menstruation and pregnancy are inflammatory conditions that cause a degree of physiological ischemia-reperfusion tissue injury, albeit much more so in pregnancy. Thus, the emergence of cyclic menstruation may not have been an evolutionary coincidence but serves to protect uterine tissues from the profound hyperinflammation and oxidative stress associated with deep placentation, a process known as preconditioning. The concept of menstrual preconditioning provides a novel paradigm for understanding how reproductive disorders impact on pregnancy outcome. For example, endometriosis could be viewed as a disorder of exaggerated menstrual preconditioning that confers protection against placentation-related disorders, such as preeclampsia. PMID- 19136086 TI - Pregnancy loss rate after mid-trimester amniocentesis in twin pregnancies. AB - OBJECTIVE: The purpose of this study was to estimate the attributable pregnancy loss rate of mid-trimester amniocentesis in twin pregnancies. STUDY DESIGN: A 16 year retrospective cohort of all twin pregnancies that underwent ultrasound evaluation at a large tertiary care medical center was followed for pregnancy outcomes. Women who underwent amniocentesis were compared with those who did not. Fetal loss was defined as loss before 24 weeks' gestation. Univariable, stratified, multivariable, and time-to-event analyses were performed. RESULTS: Of 1934 twin pregnancies, 311 women elected amniocentesis. Women who elected amniocentesis were more likely to experience a pregnancy loss than those who did not (3.2% vs 1.4%; risk difference, 1.80%; 95% CI, -0.24 to 3.84%), which was significant after adjustment for advanced maternal age, chorionicity, sonographic findings, alcohol exposure, and race (adjusted odds ratio, 2.9; 95% CI, 1.2-6.9). CONCLUSION: The attributable risk of pregnancy loss before 24 weeks' gestation after mid-trimester amniocentesis in twin pregnancies is 1 in 56 (1.8%). This information will be useful in counseling patients with twin pregnancies who are considering amniocentesis. PMID- 19136087 TI - Patient satisfaction with thermal balloon ablation for treatment of menorrhagia. AB - OBJECTIVE: The purpose of this study was to determine whether satisfaction in women who have undergone the thermal balloon ablation (TBA) procedure for menorrhagia at the Penn State Milton S. Hershey Medical Center is consistent with initial published studies. STUDY DESIGN: Two hundred sixteen patients were mailed a survey regarding patient satisfaction, postoperative bleeding patterns, and need for additional surgery. The follow-up interval was 13-60 months. RESULTS: The survey response rate was 88%. Eighty-nine percent of women were satisfied with the results of their procedure. After 3-5 years, 37% of women reported amenorrhea and 44% reported minimal/light bleeding. Only 9% of women eventually required hysterectomy. CONCLUSION: This study confirms a patient satisfaction rate in our institution that is consistent with initial published studies. PMID- 19136090 TI - Risk factors associated with postpartum suicide attempt: the chilling story. PMID- 19136088 TI - Fecal incontinence in obese women with urinary incontinence: prevalence and role of dietary fiber intake. AB - OBJECTIVE: This study estimates the prevalence of fecal incontinence (FI) in overweight and obese women with urinary incontinence and compares dietary intake in women with and without FI. STUDY DESIGN: A total of 336 incontinent and overweight women in the Program to Reduce Incontinence by Diet and Exercise clinical trial were included. FI was defined as monthly or greater loss of mucus, liquid, or solid stool. Dietary intake was quantified using the Block Food Frequency Questionnaire. RESULTS: Women had a mean (+/- SD) age of 53 +/- 10 years, body mass index of 36 +/- 6 kg/m(2), and 19% were African American. Prevalence of FI was 16% (n = 55). In multivariable analyses, FI was independently associated with low fiber intake, higher depressive symptoms, and increased urinary tract symptoms (all P < .05). CONCLUSION: Overweight and obese women report a high prevalence of monthly FI associated with low dietary fiber intake. Increasing dietary fiber may be a treatment for FI. PMID- 19136091 TI - Gestational weight gain by body mass index among US women delivering live births, 2004-2005: fueling future obesity. AB - OBJECTIVE: Current pregnancy weight gain guidelines are based on prepregnancy body mass indices (BMI), but gestational weight gains by BMI class among US women are unknown. STUDY DESIGN: We assessed the amount of gestational weight gain among 52,988 underweight, normal-weight, overweight, and obese US women who delivered a singleton, full-term infant in 2004-2005. Excessive weight gain during pregnancy was defined as gaining 35 or more pounds for normal-weight and 25 or more pounds for overweight women. RESULTS: Approximately 40% of normal weight and 60% of overweight women gained excessive weight during pregnancy. Obese women gained the least, although one-fourth of these women gained 35 or more pounds. Excessive weight gain levels were highest among women aged 19-years old or younger and those having their first birth. CONCLUSION: Excessive gestational weight gains were common, especially among the youngest and those who were nulliparous. These results predict higher obesity levels from pregnancy weight gains among US women. PMID- 19136092 TI - Indications for delivery and short-term neonatal outcomes in late preterm as compared with term births. AB - OBJECTIVE: The objective of the study was to evaluate the indications for late preterm birth and compare outcomes by gestational age among late preterm (34-36 weeks) and term (> or = 37 weeks) neonates at our institution. STUDY DESIGN: This was a retrospective analysis of delivery indications and short-term neonatal outcomes in women who delivered at the University Hospital between January 1, 2005 and Dec. 31, 2006. Data were analyzed using chi(2), Student's t-test, analysis of variance, and post hoc Tukey tests. RESULTS: One hundred forty-nine late preterm (n = 49 for 34, n = 50 for 35, n = 50 for 36 weeks) and 150 term infants (n = 50 for 37, n = 50 for 38, n = 50 for 39 weeks or longer) were evaluated. Differences among groups (ie, 34 vs 35 vs 36 vs 37, etc) as well as combinations of differences between 2 groups (ie, 34-36 weeks vs > or = 37 or > or = 38 weeks) were analyzed. Spontaneous labor and/or rupture of membranes were the most common indications for late preterm delivery (92%). Compared with term, late preterm infants had longer hospital stays (5 days vs 2.4 days; P < .001) and higher rates of neonatal intensive care unit (NICU) admissions (56% vs 4%; P < .001), feeding problems (36% vs 5%; P < .001), hyperbilirubinemia (25% vs 3%; P < .001), and respiratory complications (20% vs 5%; P < .001). Neonatal complications were minimal at 38 weeks or longer. CONCLUSION: Rates of neonatal intensive care unit admission, length of stay, and neonatal morbidities are significantly higher in late preterm as compared with term births. PMID- 19136093 TI - The changing specter of uterine rupture. AB - OBJECTIVE: The objective of the study was to review all patient records discharged with codes for uterine rupture in 2006 in Hospital Corporation of America hospitals. STUDY DESIGN: All patient charts were distributed to a committee of perinatologists and general obstetricians. Case report forms were analyzed for variables of interest to determine validity of coding and quality of care. RESULTS: Of 69 cases identified, only 41 were true ruptures. Twenty patients had previous cesareans, and in 9 of these patients, concurrent use of oxytocics was documented. Among the 21 patients without previous cesareans, 7 had previous uterine surgery, and oxytocics were documented in 12 of the remaining 14 patients. Standard of care violations were identified in 10 of 41 true rupture cases. CONCLUSION: Epidemiological data on uterine rupture based on hospital discharge codes without concurrent chart review may be invalid. Patients with previous cesareans represent only half of true uterine ruptures in contemporary practice. PMID- 19136094 TI - Laparoscopic vs vaginal hysterectomy for benign pathology. AB - OBJECTIVE: The objective of the study was to compare length of stay, blood loss, operative time, and pain of laparoscopic and vaginal hysterectomy. STUDY DESIGN: This was a prospective, randomized, controlled comparison between vaginal (VH) and laparoscopic (LH) hysterectomy among 60 consecutive patients with a uterine volume of 300 mL or less and without uterine prolapse. Patients were followed up for 12 months. RESULTS: The groups were significantly different for mean operative time (VH: 81 +/- 30 minutes; LH: 99 +/- 25 minutes; P = .033) and blood loss (LH: 83 +/- 57 mL; VH: 178 +/- 149 mL; P = .004). Bilateral adnexectomy was performed when preoperatively planned in 73% of cases of the vaginal arm, whereas it was always performed in the laparoscopic arm (P = .045). Postoperative pain on day 0 and the number of days of analgesic request were higher in the vaginal group (P = .023 and P = .017, respectively). LH was associated with a reduced hospital stay (LH: 2.7 +/- 0.5 days; VH: 3.2 +/- 0.6 days; P < .001).There were no differences between the groups at the follow-up. CONCLUSION: Laparoscopic hysterectomy results in a shorter hospital stay, less blood loss, and less postoperative pain compared with vaginal hysterectomy. PMID- 19136095 TI - Optimizing the management of stage II endometrial cancer: the role of radical hysterectomy and radiation. AB - OBJECTIVE: The optimal management of stage II endometrial cancer remains uncertain. We examined the role of radical hysterectomy and adjuvant radiotherapy for stage II endometrial cancer. STUDY DESIGN: The Surveillance, Epidemiology, and End Results database was used to identify 1577 women with stage II endometrioid type endometrial adenocarcinoma who underwent surgical staging. RESULTS: The cohort included 1198 women who underwent simple hysterectomy (76%) and 379 who underwent radical hysterectomy (24%). Radical hysterectomy had no effect on survival (hazard ratio [HR], 0.86; 95% confidence interval [CI], 0.61 1.23). Patients who did not receive radiation were 48% (HR, 1.48; 95% CI, 1.14 1.93) more likely to die than those who underwent adjuvant radiotherapy. The survival benefit from radiation was most pronounced in women who underwent radical hysterectomy. CONCLUSION: Adjuvant radiation improves survival. Although the routine performance of radical hysterectomy does not appear to be justified, patients with high-risk stage II tumors appear to benefit from combination therapy with radical hysterectomy and radiotherapy. PMID- 19136096 TI - Anal sphincter structure and function relationships in aging and fecal incontinence. AB - OBJECTIVE: The objective of the study was to determine the effect of aging and continence status on the structure and function of the external (EAS) and internal (IAS) anal sphincters STUDY DESIGN: Young (YC) and older (OC) continent women were compared with older women with fecal incontinence (OI). Patients completed the Fecal Incontinence Quality of Life and Fecal Incontinence Severity Index and underwent anorectal manometry and transanal ultrasound. RESULTS: Nine YC, 9 OC, and 8 OI women participated. Aging was associated with a thickening of the IAS. Older incontinent women had a thinner EAS, had decreased maximum squeeze pressures, and were hypersensitive to rectal distention with decreased tolerable rectal volumes and urge to defecate at lower volumes. CONCLUSION: Thickening of the IAS occurs with aging. Thinning of the EAS and a corresponding drop in squeeze pressure correlated with fecal incontinence but not aging. Rectal hypersensitivity was associated with fecal incontinence rather than aging and may play a role in the mechanism of fecal incontinence. PMID- 19136097 TI - Impaired prefrontal cortical function and disrupted adaptive cognitive control in methamphetamine abusers: a functional magnetic resonance imaging study. AB - BACKGROUND: Methamphetamine (MA) abuse is associated with neurotoxicity to frontostriatal brain regions with deleterious effects on cognitive processes. Deficits in behavioral control are thought to be one contributing factor to the sustainment of addictive behaviors in MA abuse. METHODS: In order to examine patterns of behavioral control relevant to addiction, we employed a fast-event related functional magnetic resonance imaging design to examine trial-to-trial reaction time (RT) adjustments in 12 MA-dependent subjects and 16 non-substance abusers. A variant of the Stroop task was employed to contrast the groups on error rates, RT conflict, and the level of trial-to-trial adjustments seen after incongruent trials. RESULTS: The MA abusers exhibited reduced RT adjustments and reduced activation in the right prefrontal cortex compared to controls on conditions that measured the ability to use exposure to conflict situations (i.e., conflict trials) to regulate behavior. The groups did not differ on accuracy rates or within-trial Stroop conflict effects. CONCLUSIONS: The observed deficits in trial-to-trial RT adjustments suggest that the ability to adapt a behavioral response based on prior experience may be compromised in MA abusers. These failures to modify behavior based on prior events may reflect a deficit that contributes to drug-seeking behavior. PMID- 19136098 TI - Outcome after enteroscopy for patients with obscure GI bleeding: diagnostic comparison between double-balloon endoscopy and videocapsule endoscopy. AB - BACKGROUND: Double-balloon endoscopy (DBE) and videocapsule endoscopy (VCE) have been useful in managing obscure GI bleeding (OGIB). OBJECTIVE: This study compared diagnostic yields of OGIB between DBE and VCE, and evaluated the outcome after DBE. DESIGN: A single-center retrospective study. SETTING: A tertiary referral hospital. PATIENTS: Between June 2003 and February 2007, 162 consecutive patients with OGIB were enrolled and treated. The diagnostic yield between VCE and DBE was compared in 74 patients. MAIN OUTCOME MEASUREMENTS: Comparison of diagnostic yields between DBE and VCE, and the prognosis after DBE. RESULTS: Of 162 patients, 95 (59%) were diagnosed with small-bowel diseases. They were treated by medical, enteroscopic, and surgical therapies (n = 35, 30, and 30, respectively). A comparison of the overall diagnostic yield between DBE (64%) and VCE (54%) was not significantly different. The 4 VCE-positive DBE-negative cases were because of inaccessibility of DBE. The 11 VCE-negative DBE-positive cases were because of a failure to detect lesions in the proximal small bowel and the Roux-en-Y loop, and because of diverticula. At a median follow-up of 555 days after DBE, 11 patients with small-bowel diseases developed rebleeding; all were treated by enteroscopic or medical therapies. Vascular diseases, comorbidities, especially portal hypertensive disease and chronic renal failure that required hemodialysis, and severe anemia (Hb or =5 vs <5) were associated with a higher polyp-detection rate (40% vs 24%, P < .02). BBPS scores were inversely correlated with colonoscope insertion (r = -0.16, P < .003) and withdrawal (r = -0.23, P < .001) times. LIMITATIONS: Single-center study. CONCLUSIONS: The BBPS is a valid and reliable measure of bowel preparation. It may be well suited to colonoscopy outcomes research because it reflects the colon's cleanliness during the inspection phase of the procedure. PMID- 19136103 TI - Comparison of detectability of small-bowel lesions between capsule endoscopy and double-balloon endoscopy for patients with suspected small-bowel disease. AB - BACKGROUND: There are many reports of the usefulness of capsule endoscopy (CE) or double-balloon endoscopy (DBE) for the detection of small-bowel disease. However, there are few reports that compared CE and DBE. OBJECTIVE: To determine whether CE or DBE better detects small-bowel lesions in patients with suspected small bowel disease. DESIGN: A prospective single-center study. SETTING: Department of Endoscopy, Hiroshima University Hospital. MAIN OUTCOME MEASUREMENTS: Rates of CE and DBE detection of small-bowel lesions. PATIENTS: Seventy-six consecutive patients (47 men, 29 women; mean age 56.0 years) who underwent both CE and DBE. METHODS: CE was performed before DBE. DBEs were performed within 1 week, by both retrograde and antegrade approaches so that the entire small bowel could be examined, if possible. RESULTS: Small-bowel lesions were detected by CE in 42 patients (55.3%) and by DBE in 46 patients (60.5%). The difference was not significant (P = .45). Total enteroscopy was achieved by both examinations in 35 patients, and small-bowel lesions were detected by both examinations in 21 of the 35 patients (60.0%). Agreement between results of the 2 examinations was good (kappa = 0.76). LIMITATION: The main indication for examinations was not the same. CONCLUSIONS: CE and DBE are nearly equal in their ability to detect small bowel lesions if the entire small bowel is examined. PMID- 19136104 TI - Endoscopic balloon dilation in caustic-induced chronic gastric outlet obstruction. AB - BACKGROUND: The standard treatment of caustic-induced gastric outlet obstruction (GOO) is surgery. There are only a few reports in the medical literature on endoscopic balloon dilation (EBD) for caustic-induced GOO. OBJECTIVE: To study the short-term and long-term response of EBD in patients with caustic-induced GOO. SETTING: Tertiary-care center in India. DESIGN: Retrospective analysis of data. PATIENTS: Of the 49 patients with caustic-induced GOO seen by us between January 1998 and December 2003, 41 were treated by EBD. Thirty-seven patients had consumed an acid and 4 had consumed an alkali a mean (SD) of 19.5 +/- 14.5 weeks earlier. EBD was performed every 3 weeks by using through-the-scope balloons under endoscopic guidance. INTERVENTION: The balloon was negotiated across the narrowed segment and inflated for 60 seconds by using a pressure gun. Balloons of incremental diameters, up to a maximum of 3 sizes, were used in each sitting. The end point of dilation was 15 mm, after which patients were assessed for recurrence. The patients were observed until August 2007. RESULTS: All 41 patients (23 men; mean [SD] age 29.6 +/- 8.5 years) could be successfully taken for EBD. Thirty-nine patients underwent successful repeated dilations, which required a mean (SD) of 5.8 +/- 2.6 dilations (range 2-13) to achieve the end point of 15 mm. All 39 patients were followed up for an average (SD) of 35.4 +/- 11.1 months (range 18-58 months). The mean (SD) size of the first dilator was 8.2 +/- 0.6 mm (range 8-10 mm). One patient had a perforation and was subjected to antrectomy; another patient had pain every time he received EBD; he also had surgery. Other complications were minor: self-limiting pain (n = 8) or bleeding (n = 7). CONCLUSIONS: EBD is a safe, effective, and long-lasting alternative to surgery for caustic-induced GOO. PMID- 19136105 TI - The role of postoperative colonoscopic surveillance after radical surgery for colorectal cancer: a prospective, randomized clinical study. AB - BACKGROUND: Although colonoscopy plays an important role in postoperative surveillance of patients with colorectal cancer, the optimum protocol for colonoscopic surveillance has not been established. OBJECTIVE: Our purpose was to compare the efficacy of 2 different colonoscopic surveillance strategies in terms of both survival and recurrence resectability. DESIGN: Prospective, randomized, controlled trial. SETTING: A teaching hospital in Sun Yat-sen University. PATIENTS: Three hundred twenty-six consecutive patients undergoing radical surgery for colorectal cancer. INTERVENTION: In the intensive colonoscopic surveillance group (ICS group, n = 165), colonoscopy was performed at 3-month intervals for 1 year, at 6-month intervals for the next 2 years, and once a year thereafter. In the routine colonoscopic surveillance group (RCS group, n = 161), colonoscopy was performed at 6 months, 30 months, and 60 months postoperatively. MAIN OUTCOME MEASUREMENTS AND RESULTS: The 5-year survival rate was 77% in the ICS group and 73% in the RCS group (P > .05). Postoperative colorectal cancer was detected in 13 patients (8.1%) in the ICS group and in 18 patients (11.4%) in the RCS group. In the ICS group, there were more asymptomatic postoperative colorectal cancers (P = .04), more patients had reoperation with curative intent (P = .048), and the probability of survival after postoperative colorectal cancer was higher (P = .03). LIMITATION: Lack of detailed characterization of metachronous colorectal adenomas in these patients. CONCLUSIONS: Although the patients in the ICS group had more curative operations for postoperative colorectal cancer and survived significantly longer, ICS itself did not improve overall survival. PMID- 19136106 TI - Low prevalence of submucosal invasive carcinoma at esophagectomy for high-grade dysplasia or intramucosal adenocarcinoma in Barrett's esophagus: a 20-year experience. AB - BACKGROUND: The rate of occult adenocarcinoma at esophagectomy in patients with Barrett's esophagus (BE) and high-grade dysplasia (HGD) has been reported to be approximately 40%. Recently, it has been suggested that this risk may be overestimated. OBJECTIVE: Our purpose was to determine the rate of submucosal invasive adenocarcinoma in patients undergoing esophagectomy for BE after biopsy diagnosis of HGD or intramucosal carcinoma (IMC). A secondary aim was to identify clinical risk factors for submucosal invasive adenocarcinoma in these patients. DESIGN: A retrospective study. SETTING: Tertiary referral center. PATIENTS: All patients with preoperative BE with HGD or IMC treated with esophagectomy over a 20 year period. INTERVENTIONS: Esophagectomy. MAIN OUTCOME MEASUREMENTS: Submucosal invasive adenocarcinoma at esophagectomy. RESULTS: Sixty patients were included (41 with preoperative HGD, 19 with preoperative IMC). The overall rate of submucosal invasive carcinoma was 6.7% (95% CI, 1.8%-16.2%) (n = 4), with a 5% rate of submucosal invasion in patients with preoperative HGD and 11% for patients with preoperative IMC. All 4 patients with submucosal invasion at esophagectomy had either nodular or ulcerated mucosa on preoperative endoscopy. The 1-year and 5-year all-cause risks of death for the entire cohort were 1.9% and 10.9%, respectively. LIMITATIONS: Retrospective study. CONCLUSIONS: The rate of submucosal invasive adenocarcinoma at esophagectomy in BE patients with HGD or IMC on biopsy is much lower than 40%. After adequate sampling and staging, patients with BE with HGD and IMC, especially those without endoscopically visible lesions, can potentially be treated by nonsurgical (local) therapies. PMID- 19136107 TI - EUS-assisted band ligation of small duodenal stromal tumors and follow-up by EUS. AB - BACKGROUND: Duodenal GI stromal tumors (GISTs), with potential for malignant transformation, arise in muscularis propria. It is difficult to endoscopically resect lesions in the muscularis propria by using standard electrosurgical techniques. OBJECTIVES: Our purpose was to investigate the efficacy of a new method for resection of these tumors, EUS-assisted band ligation. DESIGN: Prospective study. SETTING: Shenjing Hospital of China Medical University. PATIENTS: Nineteen elderly or high surgical risk patients with small duodenal GISTs. INTERVENTION: A standard endoscope with a transparent cap attached to the tip was used. The cap was placed over the lesion, maximum sustained suction was applied, and an elastic band was released around the base. EUS was used to determine whether the hypoechoic mass was confined completely by the band. MAIN OUTCOME MEASUREMENTS: Beginning 2 weeks after banding, the lesions were observed endoscopically once per week until healing was complete. Thereafter, all patients underwent EUS every 2 to 3 months on schedule. RESULTS: The tumors sloughed completely. The mean time required for complete healing after band ligation was 4.7 weeks. Bleeding occurred in 2 patients 7 days after ligation because the lesion sloughed. The bleeding was self-limiting and not life threatening. No perforation or other complications occurred. Follow-up ranged from 18 to 63 months, during which time no recurrence was observed. LIMITATION: New technique with limited data. CONCLUSION: EUS-assisted band ligation with systematic follow up by EUS is an effective and safe treatment for small duodenal GISTs. PMID- 19136108 TI - Metallic stents for gastric outlet obstruction: reintervention rate is lower with uncovered versus covered stents, despite similar outcomes. AB - BACKGROUND: Self-expandable metallic stents (SEMSs) are widely used for palliation of malignant gastric outlet obstruction (GOO). A common complication of their use, however, is stent obstruction caused by tumor ingrowth or hyperplasia. The covered SEMS was designed to prevent these problems. OBJECTIVE: We compared the performance of uncovered and covered SEMSs in patients with GOO. DESIGN: A retrospective study, single center. SETTING: A tertiary-referral center. PATIENTS: Sixty patients with symptomatic malignant GOO. INTERVENTIONS: All patients received an uncovered or covered knitted nitinol stent by using the over-the-wire placement procedure. MAIN OUTCOME MEASUREMENT: Comparison of the clinical outcome, complications, and the reintervention rate between uncovered and covered stents. RESULTS: Thirty-one patients (mean [+/-SEM] age 72.2 +/- 2.1 years; 16 men) received uncovered SEMSs, and 29 (mean [+/-SEM] age 70.6 +/- 1.7 years; 17 men) received covered SEMSs. The technical success rate was 100% in both groups. No difference in clinical success was seen (90.3% uncovered group vs 86.2% covered group). Regarding early complications (<1 week), one mild case of pancreatitis from the stent covering the papilla occurred in each group. Late complications included reobstruction, migration, bleeding, stent fracture, and perforation. The occurrence of reobstruction did not differ between the 2 groups (3.2% uncovered group vs 10.3% covered group). No difference in migration (0% uncovered group vs 6.9% covered group) was seen. The uncovered group required less frequent reinterventions for stent reobstruction, migration, or stent fracture (3.2% uncovered group vs 20.7% covered group, P = .0490). The uncovered group had 2 major late complications: bleeding and perforation. All 60 patients died, with a median survival time of 51 days and 62 days, respectively. LIMITATIONS: Small-sized, single-center, retrospective study. CONCLUSIONS: In palliation for malignant GOO, covered stents were associated with a more frequent need for reintervention than uncovered stents, despite similar outcomes and complications. These results require confirmation in a larger randomized comparison. PMID- 19136109 TI - A rare case of histologically mixed-type intramucosal gastric cancer accompanied by nodal recurrence and liver metastasis after endoscopic submucosal dissection. PMID- 19136110 TI - Multiple endoscopic biopsies in research subjects: safety results from a National Institutes of Health series. AB - BACKGROUND: Routine endoscopic mucosal biopsies are generally considered safe. However, the outcomes of performing large numbers of biopsies in subjects enrolled in research protocols have not been reported. OBJECTIVE: Our purpose was to assess the safety of taking numerous mucosal biopsy specimens during endoscopic procedures (eg, >20/endoscopic procedure) in research subjects. DESIGN: Single-center retrospective chart review. SETTING: Research hospital: National Institutes of Health (NIH) Clinical Center. PATIENTS: Volunteers who underwent research protocol endoscopies with large numbers of biopsies during 2001 to 2008 at the NIH. MAIN OUTCOME MEASUREMENTS: Charts were reviewed for the occurrence of procedure-related major/minor complications. RESULTS: A total of 253 research endoscopies were performed on 133 patients: 169 colonoscopies, 64 sigmoidoscopies, and 20 upper endoscopies. A total of 9,661 biopsy specimens were obtained for research and histopathologic examination (mean 38.2 +/- 15.6 per procedure). No major complications were identified. Minor complications occurred with 13 (5.1%) lower endoscopic procedures and included self-limited bleeding (4), pain (5), or both (4). There was no statistically significant association between the number of biopsies, type of procedure, location of research biopsies, operator, polypectomy, or the use of nonsteroidal anti-inflammatory drugs and the risk of complications. LIMITATIONS: Retrospective design, modest sample size. CONCLUSIONS: This is the first report on the safety of performing large numbers of endoscopic biopsies in research subjects. This practice is well tolerated and appears to have no more than minimal risk without appreciably increasing the risk of otherwise routine endoscopy. PMID- 19136111 TI - Safety and utility of ERCP during pregnancy. AB - BACKGROUND: ERCP is an important diagnostic and therapeutic tool in patients with biliary and pancreatic disease. Its utility and safety during pregnancy is largely unknown because it is not often required and because its use has been only infrequently reported in the published literature. OBJECTIVE: Our purpose was to report the clinical experience with ERCP during pregnancy. DESIGN: Retrospective review, single academic center. PATIENTS: All (consecutive) pregnant women who underwent ERCP at Parkland Memorial Hospital from 2000 to 2006. MAIN OUTCOME MEASUREMENTS: History, clinical data, hospital course, procedure-related complication rates and outcomes, and delivery and fetal outcomes were abstracted from medical records. RESULTS: During the study period, 68 ERCPs were performed on 65 pregnant women. The calculated ERCP rate was 1 per 1415 births. The common indications for ERCP in pregnancy were recurrent biliary colic, abnormal liver function tests, and dilated bile duct on US. ERCP was technically successful in all patients. The median fluoroscopy time was 1.45 minutes (range 0-7.2 minutes). There was no perforation, sedation-related adverse event, postsphincterotomy bleeding, cholangitis, or procedure-related maternal or fetal deaths. Post-ERCP pancreatitis was diagnosed in 11 patients (16%). None of these 11 patients had local or systemic complications. Fifty-nine patients had complete follow-up. Endoscopic therapy at the time of ERCP was undertaken in all patients. Furthermore, 9 patients (32.1%) underwent cholecystectomy in the first and second trimesters for either acute cholecystitis (6) or symptomatic gallstones (3). Term pregnancy was achieved in 53 patients (89.8%). Patients having ERCP in the first trimester had the lowest percentage of term pregnancy (73.3%) and the highest risk of preterm delivery (20.0%) and low-birth-weight newborns (21.4%). None of the 59 patients with long-term follow-up had spontaneous fetal loss, perinatal death, stillbirth, or fetal malformation. LIMITATION: Retrospective review. CONCLUSIONS: ERCP can be performed safely during pregnancy. Further, ERCP performed in pregnancy leads to specific therapy in essentially all patients. However, ERCP may be associated with a higher rate of post-ERCP pancreatitis than in the general population. PMID- 19136112 TI - A transformation method to estimate muscle attachments based on three bony landmarks. AB - In order to create musculoskeletal models that can be scalable to different subject specificities the calculation of the exact locations of muscle attachment is required. For this purpose, a scaling method is presented that estimates muscle attachment locations in homologous segments using three bony landmarks per segment. A data-set of 17 muscles' attachment lines from the shoulders of seven cadavers was used to assess the estimation quality of the scaling method. By knowing from the cadaver data the measured location of the muscles' attachment lines it is possible to assess the quality of the estimated ones. The scaling results showed an overall mean RMSE for the scapula and humerus muscles of 7.6 and 11.1mm, respectively. These results were then analyzed with an upper extremity model, in order to compute the influence of the RMSE in glenohumeral elevation muscle moment arms in the scapular plane. The results presented were considered to be satisfactory. Among other error contributors, the inter- and intra-subject variability should be further investigated, along with the sensitivity of a biomechanical model to these error variations. PMID- 19136113 TI - Influence of single-level lumbar degenerative disc disease on the behavior of the adjacent segments--a finite element model study. AB - The current study investigated mechanical predictors for the development of adjacent disc degeneration. A 3-D finite element model of a lumbar spine was modified to simulate two grades of degeneration at the L4-L5 disc. Degeneration was modeled by changes in geometry and material properties. All models were subjected to follower preloads of 800N and moment loads in the three principal directions of motion using a hybrid protocol. Degeneration caused changes in the loading and motion patterns of the segments above and below the degenerated disc. At the level (L3-L4) above the degenerated disc, the motion increased due to moderate degeneration by 21% under lateral bending, 26% under axial rotation and 28% under flexion/extension. At the level (L5-S1) below the degenerated disc, motion increased only during lateral bending by 20% due to moderate degeneration. Both the L3-L4 and L5-S1 segment showed a monotonic increase in both the maximum von Mises stress and shear stress in the annulus as degeneration progressed for all loading directions, expect extension at L3-L4. The most significant increase in stress was observed at the L5-S1 level during axial rotation with nearly a ten fold increase in the maximum shear stress and 103% increase in the maximum von Mises stress. The L5-S1 segment also showed a progressive increase in facet contact force for all loading directions with degeneration. Nucleus pressure did not increase significantly for any loading direction at either the caudal or cephalic adjacent segment. Results suggest that single-level degeneration can increase the risk for injury at the adjacent levels. PMID- 19136114 TI - Design and validation of a dynamometric horseshoe for the measurement of three dimensional ground reaction force on a moving horse. AB - Properties of ground surfaces condition locomotion, and quality of track surfaces is believed to be involved in the pathogenesis of many musculoskeletal injuries in the horse. Measuring ground reaction forces (GRF) is an interesting approach to assess those interactions. Forceplates are the most commonly used but they are not well suited to compare different ground surfaces at fast gaits. Embarked equipment, fixed under the horse's hoof, would allow force measurement on any track. The objective of this work was (1) to design a device which enables the measurement of 3-D GRF on any ground, at any gait, for a given subject, (2) to determine its accuracy, and (3) to evaluate its performance and usefulness under physiological conditions. The resulting dynamometric horseshoe was composed of 4 piezoelectric sensors sandwiched between 2 aluminium plates designed at the shape of an equine shoe. The measurements, evaluated after a quasi-static calibration, revealed that the root mean square error was 1.3% in the normal direction, and 3.1% in the transversal direction. In vivo tests at the walk and trot in straight line and at the trot on circles, were conducted on 3 different ground surfaces. The results demonstrate that this dynamometric horseshoe is well suited to study the effects of different ground surfaces on GRF in the moving horse. PMID- 19136115 TI - Separation of proanthocyanidins isolated from tea leaves using high-speed counter current chromatography. AB - The proanthocyanidin extract from tea (Camellia sinensis) leaves was purified for the further study of the biological role of proanthocyanidins in blister blight leaf disease of tea, which is caused by the fungus Exobasidium vexans. An aqueous acetone extract of proanthocyanidins prepared from healthy tea leaves was partially purified using Sephadex LH-20 chromatography. The crude proanthocyanidin extract obtained was fractionated with high-speed counter current chromatography (HSCCC) using the solvent system n-hexane-EtOAc-MeOH-water (1:5:1:5). The purity of the each isolated fraction after a single HSCCC run was evaluated by high-performance liquid chromatography (HPLC). Seven fractions of high purity were isolated. The identity of the compound present in each fraction isolated was established using electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy. Five proanthocyanidins and two flavanol digallates, (-)-epigallocatechin digallate (EGCDG) and (-)-epicatechin digallate (ECDG) were isolated. Comparison of spectral data of the proanthocyanidins isolated with those previously reported indicated that all five were known B-type proanthocyanidins with 2,3-cis stereochemistry in both the upper (u-unit) and the terminal (t-unit) units, and 4R configuration of the C ring in the u-unit. The proanthocyanidins were established to be dimers composed of (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and (-) epiafzelechin gallate (EAG) units with the following structures: EGCG-(4beta-->6) EGCG, ECG-(4beta-->6)-EGCG, EGCG-(4beta-->6)-ECG, EAG-(4beta-->6)-EGCG, ECG (4beta-->6)-ECG by analysis of spectral data. Therefore HSCCC offers a powerful method for the separation of a group of closely related naturally occurring compounds. PMID- 19136116 TI - Statistical theory of multiple-site linear wall-adsorption capillary Chromatography. AB - Based on the mass-balance principle, a particular diffusion equation to describe the movement of solute molecules in the stagnant layer of multiple-site solid surfaces is constructed. From the equation, the moments of residence time in a step on multiple-site surfaces are derived. Similarly, the moments in a step in the mobile phase are also derived from a diffusion-drift equation. According to the probability theory, there exists a general relationship between the moments of an elution curve and the moments in a step. Through this relationship, the expressions of the elution-curve moments are derived from the step moments. In this paper, the details related to multiple-site linear wall-adsorption capillary chromatography are described and added in the equations to determine the step moments. The resultant expressions of the elution-curve moments involve various factors, such as adsorption-desorption rate constants, equilibrium constants, axial and radial dispersions in the mobile phase. Afterwards, the moment expressions are used to analyze the peak tailing. The results show that a small quantity of sites with a slow desorption rate will lead to a large peak asymmetry. PMID- 19136117 TI - Liposome rupture and contents release over coplanar microelectrode arrays. AB - The vulnerability of vesicles to electroporation and rupture by externally applied electric fields, combined with the ability of dielectrophoresis and/or AC electroosmosis to manipulate suspended vesicles over micropatterned electrodes suggests new techniques to electrically trigger localized chemical reactions at predetermined positions in microfluidic devices. The electric field conditions needed to rupture giant unilamellar phospholipid vesicles were determined as a function of vesicle size in a simple coplanar microelectrode array geometry. Rupture results were interpreted in terms of the spatially varying electric field strength, calculated via the Poisson equation and accounting for frequency effects on electrode impedance, and the experimentally measured vesicle elevation. The vesicle transmembrane voltage scales linearly with the applied electric field strength according to the Schwan theory of electroporation, so that larger vesicles are usually more prone to electric field induced rupture than smaller ones in the uniform electric fields that are typically employed to cause electroporation and rupture. Yet, in the coplanar microelectrode arrangement, larger vesicles preferentially reside at larger elevations where the local field strengths are weaker. As a result, there is a sensitive range of vesicle radii that are most prone to electric field induced rupture over a micropatterned electrode array that leaves the largest vesicles resistant to rupture. PMID- 19136118 TI - A facile route to ultra-long polyaniline nanowires and the fabrication of photoswitch. AB - Ultra-long polyaniline nanowires with an average diameter of 60 nm and length up to tens of micrometers were successfully synthesized via chemical oxidation polymerization in an aqueous solution. These nanowires exhibited reversible electrochemical behavior judged from cyclic voltammetry curves. The excellent photosensitivity and photoresponse of a bundle of nanowires were also investigated, which showed that the photocurrent enhanced by ca. 4 times under irradiation of an incandescence lamp (12 V, 10 W). This work might be useful in the fabrication of photosensor and photoswitch nanodevices in the future. PMID- 19136119 TI - Efficient preparation of silver nanoplates assisted by non-polar solvents. AB - In the paper, we report an efficient method to prepare high yield (up to 97%) of silver nanoplates. Synthesis of silver nanoplates was carried out in a binary solvent system of N,N-dimethylformamide (DMF) and toluene, in which DMF served as the reductant and polyvinylpyrrolidone (PVP) as the capping agent. By increasing the ratio of toluene to DMF to 7:6, silver nanoplates can be successfully synthesized; otherwise other shaped nanoparticles would be the major products. The nanoplate sample was characterized by TEM, HRTEM, SAED, XRD, AFM and UV visible spectroscopy, proving the high nanoplate purity of this sample. The influence of toluene content, other solvents, AgNO(3) concentration, preparation temperature and chloride ions was also examined, which suggests that the function of non-polar solvents in this system is to enhance the PVP coverage on silver surface and, furthermore, to facilitate the preferential adsorption of PVP on two (111) facets of silver nanoplates. PMID- 19136120 TI - A liquid/liquid interface excited by stimulation with water. AB - An immiscible interface, which was composed of CaCl(2) aqueous and ester phases, was excited by the addition of a small amount of water. The excitation, i.e., burst and flow at the interface, was observed above a critical concentration of CaCl(2) in the aqueous phase. The critical concentration and degree of excitation depended on the kind of ester. The mechanism of excitation is discussed in relation to the interfacial tension and the Marangoni effect. PMID- 19136121 TI - Behavior of fluorescent molecules bound to the interior of silica nanocapsules in various solvents. AB - Porous silica nanocapsules with 20% 3-aminopropyltrimethoxysilane (APS)-bound 6 carboxy-fluorescein (APS-fluorescein) and 80% APS molecules adsorbed on the surface of a 50-nm-diameter Au core were prepared by a modified core-shell method. Silica mesoporous nanocapsules were obtained after the Au cores were dissolved in sodium cyanide. The size of the pores in the silica shells corresponded to the area of the fluorescein (approximately 1.02 nm(2)) in each APS-fluorescein molecule, which was bound to the silica shell by coupling between the silanol groups of APS in the APS-fluorescein molecule and the silica shell. The amino group of APS bound to the silica inside the shell is also reactive. Dy485XL N-hydroxysuccinimide ester (NHS) molecules were then added to the mesoporous silica nanocapsules in the solution and bonded to the amino group of the interior. Thus, mesoporous (fluorescein and Dy485XL)-bound silica nanocapsules were obtained. The fluorescence of Dy485XL was only observed in the mesoporous (fluorescein and Dy485XL)-bound silica nanocapsules in aqueous solution after ultrafiltration. However, the fluorescence of fluorescein reappeared after the addition of acetonitrile. Furthermore, upon adding various solvents to the mesoporous (fluorescein and Dy485XL)-bound silica nanocapsules, their fluorescence varied with that of fluorescein or Dy485XL. In the case of a mixture of 6-carboxy-fluorescein-N-hydroxysuccinimide (FLUOS) and Dy485XL-NHS free molecules in aqueous solution, the fluorescence of FLUOS was observed. Such different fluorescence phenomena demonstrated that Dy485XL-NHS molecules can easily penetrate into the nanocapsule interior via the pores and that the interior of the silica nanocapsules can bind to Dy485XL molecules. These fluorescence behaviors are discussed in terms of fluorescence resonance energy transfer (FRET) and solvatochromism. PMID- 19136122 TI - Temporal and spatial changes in marine benthic habitats in relation to the EU Water Framework Directive: the use of sediment profile imagery. AB - In 2002 to 2006, sediment profile imagery (SPI) was used to study the environmental impact of eutrophication-induced irregular and seasonal hypoxia on marine benthic habitats in six regions in the Skagerrak and Kattegat (West Sweden). The benthic habitat quality (BHQ) was assessed by parameterisation of biogenic structures observed by the SPI technique, and benthic quality status was related to the EU Water Framework Directive (EU-WFD). The temporal changes were analysed by a 5-factor nested ANOVA and significant temporal differences were recorded within three of the regions. Two of these were affected by hypoxia in the deeper parts and one was probably affected by hypoxia below the halocline. The environmental quality status according to the EU-WFD was bad to high in two regions, moderate to good in three regions, and good to high, i.e., acceptable according to the EU-WFD, in only one region. As BHQ is highly correlated to benthic faunal data, measures have to be taken to improve the coastal water quality in five of the six studied areas. PMID- 19136123 TI - Calibration and evaluation of five indicators of benthic community condition in two California bay and estuary habitats. AB - Many types of indices have been developed to assess benthic invertebrate community condition, but there have been few studies evaluating the relative performance of different index approaches. Here we calibrate and compare the performance of five indices: the Benthic Response Index (BRI), Benthic Quality Index (BQI), Relative Benthic Index (RBI), River Invertebrate Prediction and Classification System (RIVPACS), and the Index of Biotic Integrity (IBI). We also examine whether index performance improves when the different indices, which rely on measurement of different properties, are used in combination. The five indices were calibrated for two geographies using 238 samples from southern California marine bays and 125 samples from polyhaline San Francisco Bay. Index performance was evaluated by comparing index assessments of 35 sites to the best professional judgment of nine benthic experts. None of the individual indices performed as well as the average expert in ranking sample condition or evaluating whether benthic assemblages exhibited evidence of disturbance. However, several index combinations outperformed the average expert. When results from both habitats were combined, two four-index combinations and a three-index combination performed best. However, performance differences among several combinations were small enough that factors such as logistics can also become a consideration in index selection. PMID- 19136124 TI - Two poplar methyl salicylate esterases display comparable biochemical properties but divergent expression patterns. AB - Two genes encoding proteins of 98% sequence identity that are highly homologous to tobacco methyl salicylate (MeSA) esterase (SABP2) were identified and cloned from poplar. Proteins encoded by these two genes displayed specific esterase activities towards MeSA to produce salicylic acid, and are named PtSABP2-1 and PtSABP2-2, respectively. Recombinant PtSABP2-1 and PtSABP2-2 exhibited apparent Km values of 68.2+/-3.8microM and 24.6+/-1microM with MeSA, respectively. Structural modeling using the three-dimensional structure of tobacco SABP2 as a template indicated that the active sites of PtSABP2-1 and PtSABP2-2 were highly similar to that of tobacco SABP2. Under normal growing conditions, PtSABP2-1 showed the highest level of expression in leaves and PtSABP2-2 was most highly expressed in roots. In leaf tissues of poplar plants under stress conditions, the expression of PtSABP2-1 was significantly down-regulated by two stress factors, whereas the expression of PtSABP2-2 was significantly up-regulated by four stress factors. The plausible mechanisms leading to these two highly homologous MeSA esterase genes involved in divergent biological processes in poplar are discussed. PMID- 19136125 TI - Bacterial resistance modifying tetrasaccharide agents from Ipomoea murucoides. AB - As part of an ongoing project to identify oligosaccharides which modulate bacterial multidrug resistance, the CHCl(3)-soluble extract from flowers of a Mexican arborescent morning glory, Ipomoea murucoides, through preparative-scale recycling HPLC, yielded five lipophilic tetrasaccharide inhibitors of Staphylococcusaureus multidrug efflux pumps, murucoidins XII-XVI (1-5). The macrocyclic lactone-type structures for these linear hetero-tetraglycoside derivatives of jalapinolic acid were established by spectroscopic methods. These compounds were tested for in vitro antibacterial and resistance modifying activity against strains of Staphylococcus aureus possessing multidrug resistance efflux mechanisms. Only murucoidin XIV (3) displayed antimicrobial activity against SA-1199B (MIC 32microg/ml), a norfloxacin-resistant strain that over expresses the NorA MDR efflux pump. The four microbiologically inactive (MIC>512microg/ml) tetrasaccharides increased norfloxacin susceptibility of this strain by 4-fold (8microg/ml from 32microg/ml) at concentrations of 25microg/ml, while murucoidin XIV (3) exerted the same potentiation effect at a concentration of 5microg/ml. PMID- 19136126 TI - The role of tryptophan as a biosynthetic precursor of indole-diterpenoid fungal metabolites: continuing a debate. AB - Studies in the 1980s and 1990s on the origin of the indole moiety in fungal indole-diterpenoids using (14)C-labelled tryptophan consistently showed autoradiographic evidence but gave low % incorporation of the probe. Recent studies on a member of the group (nodulisporic acid A), using more specific (13)C methodology, demonstrated a role of the tryptophan biosynthetic pathway but, in failing to show involvement of end-product, concluded that the indole was derived from indole-3-glycerol phosphate and suggested that the previous (14)C data arose via metabolic scrambling of label. In considering the protocol for the (13)C studies, there is concern that the fungal material was starved of an exogenous nitrogen source and thus could have degraded added labelled tryptophan. Consequently, synthesis of the serine necessary for anabolic formation of tryptophan may have been constrained. It is suggested that (13)C studies on appropriate fungi early in the idiophase of submerged or surface fermentation should be made before the biosynthesis of indole-diterpenoids can become clearer. PMID- 19136127 TI - Synthesis and cytotoxicity evaluation of natural alpha-bisabolol beta-D fucopyranoside and analogues. AB - alpha-Bisabolol beta-d-fucopyranoside, a cytotoxic naturally occurring compound, was efficiently synthesized along with five other alpha-bisabolol glycosides (beta-d-glucoside, beta-d-galactoside, alpha-d-mannoside, beta-d-xyloside and alpha-l-rhamnoside). Glycosidation of alpha-bisabolol was performed using Schmidt's inverse procedure and provided excellent yields (83-95%). Cytotoxicity was evaluated against a broad panel of cancerous cell lines including human and rat glioma (U-87, U-251 and GL-261) since the anticancer activity of alpha bisabolol was previously demonstrated against brain tumor cell lines. The addition of a sugar moiety markedly increased alpha-bisabolol cytotoxicity in most cases. Among the synthesized glycosides, alpha-bisabolol alpha-l rhamnopyranoside exhibited the strongest cytotoxic activity with IC(50) ranging from 40 to 64muM. According to ADME in silico predictions, this glycoside closely respects physicochemical parameters necessary to cross the blood-brain barrier passively. PMID- 19136128 TI - Khayanolides from African mahogany Khaya senegalensis (Meliaceae): A revision. AB - Five khayanolides (1-O-acetylkhayanolide B 1, khayanolide B 2, khayanolide E 3, 1 O-deacetylkhayanolide E 4, 6-dehydroxylkhayanolide E 5) were isolated from the stem bark of African mahogany Khaya senegalensis (Meliaceae). Their structures and absolute configurations were determined through extensive spectroscopic analyses including MS, NMR, and single-crystal X-ray diffraction experiments. The results established that two previously reported khayanolides, 1alpha-acetoxy 2beta,3alpha,6,8alpha,14beta-pentahydroxy-[4.2.1(10,30).1(1,4)]-tricyclomeliac-7 oate 6 and 1alpha,2beta,3alpha,6,8alpha,14beta-hexahydroxy-[4.2.1(10,30).1(1,4)] tricyclomeliac-7-oate 7, were, in fact, 1-O-acetylkhayanolide B 1 and khayanolide B 2, and that the two reported phragmalin derivatives, methyl 1alpha-acetoxy 6,8alpha,14beta,30beta-tetrahydroxy-3-oxo-[3.3.1(10,2).1(1,4)]-tricyclomeliac-7 oate 8 and methyl 1alpha,6,8alpha,14beta,30beta-pentahydroxy-3-oxo [3.3.1(10,2).1(1,4)]-tricyclomeliac-7-oate 9, were, in fact, khayanolide E 3 and 1-O-deacetylkhayanolide E 4, respectively. Based on the results from this study and consideration of the biogenetic pathway, the methyl 6-hydroxyangolensate in African mahogany K. senegalensis should have a C-6 S configuration while methyl 6 hydroxyangolensate in genuine mahogany Swietenia species should have a C-6 R configuration. PMID- 19136129 TI - CD36 T188G gene polymorphism and severe falciparum malaria in India. AB - Sequestration of parasitized erythrocytes (PE) in the microvasculature contributes directly to the virulence and severe pathology of Plasmodium falciparum malaria. The scavenger receptor CD36 appears to play an important role in PE adherence. Recently several mutations in the CD36 gene have been found to be associated with variability in susceptibility to P. falciparum infection in different ethnic populations. We investigated the possible association of T188G CD36 gene polymorphism with severe clinical manifestations of malaria in 95 adult patients with severe malaria admitted to SCB Medical College Hospital, Cuttack, Orissa, India ('severe' group) and 95 ethnically matched controls attending outpatient clinics at primary health centres ('mild' group). The frequency of the wild-type (T188T) allele of the CD36 gene was 0.91 in the 'severe' group and 0.78 in the 'mild' group of patients, while mutant (T188G) allele frequency was 0.09 in the severe group and 0.22 in the mild group. The Hardy-Weinberg equation indicates that the mutant allele is under selection pressure and disease association analysis shows that the presence of the heterozygote mutant allele renders protection against severe malaria (chi(2)=10.67, odds ratio=3.51, 95% CI 1.67-7.36). PMID- 19136130 TI - Skin disorders and disease profile of poverty: analysis of medical records in Tigray, northern Ethiopia, 2005-2007. AB - Skin disease is among the most frequent causes of morbidity in Ethiopia, showing high prevalence in the general population and being the sixth most frequent cause of outpatient visits nationwide to health facilities. This study was aimed at describing skin disease patterns in Tigray region, northern Ethiopia through a retrospective analysis of 30618 outpatient and 1103 inpatient medical records from the Italian Dermatological Centre in Mekelle, the regional capital of Tigray, during the period 2005-2007. The leading causes of outpatient attendance were eczema (n=6998), mycosis (n=5065), pigmentation anomalies (n=3319), scabies (n=2229) and acne (n=2001). Different patterns were observed for inpatient services, with scabies being the leading cause of admission (n=213), followed by eczema (n=158), pyoderma (n=131), leishmaniasis (n=106) and mycosis (n=56). Since the most common and readily treatable skin diseases are related to household crowding and lack of hygiene, i.e. conditions reflecting low socio-economic status, they are considered to be important contributors to the 'disease profile of poverty' and, in general, to health inequalities. Cost-effective interventions are available to reduce the burden of skin disease. The control of skin disease should be considered a public health priority and included in strategies for health-sector development and poverty reduction. PMID- 19136131 TI - Melanogenesis in dermal melanocytes of Japanese Silky chicken embryos. AB - The Japanese Silky chicken (SK) shows dermal and visceral hyperpigmentation. This study characterizes ultrastructurally the melanin granules developing in dermal melanocytes of the dorsal skin of SK, in an attempt to better understand the processes of melanogenesis in these permanently ectopic cells. The steps of melanogenesis are similar to those described for epidermal melanocytes, with melanosomes going from stage I to IV but, in SK, the maturation occurs in the cell body, as well as in the cytoplasmic processes. At stage III, the deposition of melanin is cumulative and can aggregate in rounded structures, which combine to turn into the mature granule. The final destiny of mature melanosomes is still unclear, although it was observed that dermal macrophages can accumulate melanin granules in their phagosomes. Even with the close proximity between melanocytes and other dermal cells, the transference of melanosomes was not observed. Our findings indicate that melanogenesis in dermal melanocytes in SK has the same morphological characteristics found in epidermal melanocytes, but the functional aspect still remains to be elucidated. PMID- 19136132 TI - The kinase activity of pseudorabies virus US3 is required for modulation of the actin cytoskeleton. AB - Different viruses exploit the host cytoskeleton to facilitate replication and spread. The conserved US3 protein of the alphaherpesvirus pseudorabies virus induces actin stress fiber disassembly and formation of actin-containing cell projections, which are associated with enhanced intercellular virus spread. Proteins of members of other virus families, notably vaccinia virus F11L protein and human immunodeficiency virus Nef protein, induce actin rearrangements that are very similar to those induced by US3. Interestingly, unlike F11L and Nef, the US3 protein displays serine/threonine kinase activity. Here, we report that the kinase activity of pseudorabies virus US3 is absolutely required for its actin modulating activity. These data show that different viruses have developed independent mechanisms to induce very similar actin rearrangements. PMID- 19136133 TI - Betanodavirus non-structural protein B1: A novel anti-necrotic death factor that modulates cell death in early replication cycle in fish cells. AB - The functions of the Betanodavirus non-structural protein B1 is still unknown. We examined B1 expression patterns and investigated novel cell death regulatory functions for this viral protein following RGNNV infection in fish cells. The B1 gene (336 nt) was cloned from the redspotted grouper nervous necrosis virus (RGNNV) genome. B1 mRNA was rapidly expressed in the fish cells from viral RNA3 at 12 h post-infection (p.i.). At the protein level, expression was low at 12 h p.i., and then increased rapidly between 24 h and 72 h p.i. In RGNNV-infected, B1 containing fish cells, over expression of RGNNV B1 reduced Annexin-V positive cells by 50% and 65% at 48 h and 72 h p.i., respectively, and decreased loss of mitochondrial membrane potential (MMP) by 20% and 70% at 48 h and 72 h p.i., respectively. Finally, B1 knockdown during RGNNV infection using anti-sense RNA increased necrotic cell death and reduced cell viability during the early replication cycle (24 h p.i.). Our results suggest that B1 is an early expression protein that has an anti-necrotic cell death function which reduces the MMP loss and enhances viral host cell viability. This finding provides new insights into RNA viral pathogenesis and disease control. PMID- 19136134 TI - Small DNA tumor viruses: large contributors to biomedical sciences. AB - Studies of the small DNA tumor viruses (the polyomaviruses, the adenoviruses and the papillomaviruses) have led to fundamental discoveries that have advanced our understanding of basic mammalian cell molecular biology processes such as transcription and DNA replication, uncovered pathways and genes often perturbed in human cancer, and identified bona fide human cancer viruses. In this article we examine the many contributions that have come from the small DNA tumor virus field and provide a recounting of some of the major landmark discoveries. PMID- 19136135 TI - Achieving nitrogen removal via nitrite in a pilot-scale continuous pre denitrification plant. AB - Nitrogen removal via nitrite (the nitrite pathway) is beneficial for carbon limited biological wastewater treatment plants. However, partial nitrification to nitrite has proven difficult in continuous processes treating domestic wastewater. The nitrite pathway is achieved in this study in a pilot-scale continuous pre-denitrification plant (V=300 L) treating domestic wastewater by controlling the dissolved oxygen (DO) concentration at 0.4-0.7 mg/L. It is demonstrated that the nitrite pathway could be repeatedly and reliably achieved, with over 95% of the oxidized nitrogen compounds at the end of the aerobic zone being nitrite. The nitrite pathway improved the total nitrogen (TN) removal by about 20% in comparison to the nitrate pathway, and also reduced aeration costs by 24%. FISH analysis showed that the nitrite oxidizing bacteria (NOB) population gradually reduced at low DO levels, and reached negligible levels when stable nitrite pathway was established. It is hypothesized that NOB was washed out due to its relatively lower affinity with oxygen. A lag phase was observed in the establishment of the nitrite pathway. Several sludge ages were required for the onset of the nitrite pathway after the application of low DO levels. However, nitrite accumulation increased rapidly after that. A similar lag phase was observed for the upset of the nitrite pathway when a DO concentration of 2-3 mg/L was applied. The nitrite pathway negatively impacted on the sludge settleability. A strong correlation between the sludge volume index and the degree of nitrite accumulation was observed. PMID- 19136136 TI - The valuation of water quality: effects of mixing different drinking water qualities. AB - As water supplies increasingly turn to use desalination technologies it becomes relevant to consider the options for remineralization and blending with mineral rich water resources. We present a method for analyzing economic consequences due to changes in drinking water mineral content. Included impacts are cardiovascular diseases, dental caries, atopic eczema, lifetime of dish and clothes washing machines, heat exchangers, distribution systems, bottled water consumption and soap usage. The method includes an uncertainty assessment that ranks the impacts having the highest influence on the result and associated uncertainty. Effects are calculated for a scenario where 50% of Copenhagen's water supply is substituted by desalinated water. Without remineralization the total impact is expected to be negative (euro -0.44+/-0.2/m(3)) and individual impacts expected in the range of euro 0.01-0.51/m(3) delivered water. Health impacts have the highest contribution to impact size and uncertainty. With remineralization it is possible to reduce several negative impacts and the total impact is expected to be positive (euro 0.14+/-0.08/m(3)). PMID- 19136137 TI - Measuring the trophic status of ponds: relationships between summer rate of periphytic net primary productivity and water physico-chemistry. AB - The monitoring of the water quality is a major environmental concern and specific method of assessment needs to be addressed for ponds. Among threats on freshwater, eutrophication remains an important and widespread problem. To assess if the trophic state of ponds can be significantly predicted by physico-chemical surrogates, parallel measurements of water physico-chemistry and of summer rates of periphytic net primary productivity (NPP) were performed on a set of nine lowland Swiss ponds representing the whole spectrum of trophic states. The developed methods to measure periphytic NPP on artificial substrates give similar values between the pseudo-replicates and are functional for ponds. The winter concentration of total nitrogen (TN) in the water is a significant predictor of the average summer rates of NPP while in submerged macrophyte beds total phosphorus (TP) is a better predictor. Rates of NPP measured in open water were most often higher and water transparency lower than in macrophyte beds, highlighting the positive influence of macrophytes on water quality and a possible co-occurrence in a pond of contrasting conditions when submerged species are present. This emphasizes the importance of maintaining or restoring submerged macrophyte stands in ponds to improve the water quality. PMID- 19136138 TI - Zero Nuisance Piggeries: long-term performance of MBR (membrane bioreactor) for dilute swine wastewater treatment using submerged membrane bioreactor in semi industrial scale. AB - Effective aerobic/anoxic treatment of piggery manure wastewater was achieved in a real farm scale using a small piggery (72 pigs) with reuse of the treated water. The experimental procedure was followed for 9 months. Fresh manure (FM) is formed by daily flush on piggeries and biologically treated after centrifuge pre treatment. For upgrade liquid/solid separation and pathogen retention in biological treatment, a membrane system was used with the aim of effluent reuse in flush. Despite an evolution of FM through time, centrifuge pre-treatment and bioreactor performances stayed at high level. An elimination of 86% of the suspended solids occurred through pre-treatment, and nitrogen and COD biological degradation remains at 90% all time long. Moreover, interestingly about half of the soluble part of phosphorus (20% of the global phosphorus content) was biologically removed via the recirculation between the anoxic and the aerobic tank which acted as an intermittent aerobic/anoxic sequence. A part of COD was proved not biodegradable and was accumulated via the reuse of the treated water for flushing purpose. This accumulation justifies washing of the biomass between two runs in purpose to enhance the treated water quality and also to meet the membrane tolerance. The membrane was proved reliable as far as the maintenance procedure was respected. Maintenance cleaning had to be operated as soon as the TransMembrane Pressure (TMP) achieved 50 mbar and curative washing was necessary if the TMP increased over 90 mbar or between 2 runs. The temperature was proved to influence both the bioactivity and the membrane fouling kinetic. Finally, it was demonstrated that the process was sustainable for long-term management of swine wastewater at semi-industrial scale. PMID- 19136139 TI - Determination and prediction of xenoestrogens by recombinant yeast-based assay and QSAR. AB - Estrogenic activities expressed by the logarithm of relative potency (logRP), for 8 xenoestrogens were determined using the recombinant yeast-based assay. The determined logRP values were employed as an independent external data set to validate an estrogenic activity quantitative structure-activity relationship (QSAR) model. The QSAR model was established using partial least squares regression and molecular descriptors derived from DRAGON software. For the training set of the QSAR model that included 25 xenoestrogens, R(2)=0.889, the leave-one-out cross-validation squared correlation coefficient (Q(LOO)(2)) was 0.897. For the external validation set, the predicted logRP values were consistent with the observed values, with a root mean square error (RMSE) of 0.736 log units and the squared correlation coefficient (Q(EXT)(2)) was 0.775. Six descriptors were included in the QSAR model, which indicated that the logRP value was related to molecular size, shape profiles, symmetry and polarizability. Comparatively, the developed model has good robustness and predictivity. Moreover, the applicability domain of the model was discussed. PMID- 19136140 TI - Interactions between estrogenic chemicals in binary mixtures investigated using vitellogenin induction and factorial analysis. AB - Considerable progress has been made in assessing the combined effects of chemicals, but the effect of mixtures remains one of the most daunting challenges in environmental toxicology. In this study, the effects of binary mixtures of estrogenic chemicals were investigated using plasma vitellogenin (VTG) induction in male adult Japanese medaka (Oryzias latipes) as the endpoint. We focused on whether or not the factorial design using rigorous statistical methods was appropriate for the identification of possible interactions. The estrogenic activities of 17beta-estradiol (E2), 4-tert-nonylphenol (NP) and bisphenol A (BPA) were investigated in medaka following 14 d of exposure. Although all chemicals induced concentration-dependent increases in plasma VTG, a complete concentration-response curve was obtained only for E2, while partial curves were obtained for NP and BPA. Therefore, a 3 x 3 factorial design was employed to identify both the individual and interactive effects. A significant difference in mean plasma VTG induction was found when any chemical was considered alone. However, no significant interactions between E2 and NP, E2 and BPA, and NP and BPA were demonstrated. These results suggest that these three model estrogenic chemicals share a common mechanism for inducing VTG synthesis, and that no interactions occur when they act in combination. We also demonstrated the ability of this experimental design to detect interactions between binary mixtures, which will allow the assessment of biological effects of more complex mixtures in future research. PMID- 19136141 TI - Hydrogen peroxide decomposition on manganese oxide (pyrolusite): kinetics, intermediates, and mechanism. AB - The objective of this study is the kinetic interpretation of hydrogen peroxide decomposition on manganese oxide (pyrolusite) and the explanation of the reaction mechanism including the hydroperoxide/superoxide anion. The decomposition of hydrogen peroxide on manganese oxide at pH 7 was represented by a pseudo first order model. The maximum value of the observed first-order rates constants (k(obs)) was 0.741 min(-1) at 11.8 of [H(2)O(2)]/[triple bond MnO(2)] when [H(2)O(2)]/[triple bond MnO(2)] were ranged from 58.8 to 3.92. The pseudo first order rate constants (kMnO(2)) approximated as the average value of 0.025 (min mM)(-1) with a standard deviation of 0.003 at [H(2)O(2)]/[triple bond MnO(2)] ranged from 39.2 to 11.8. When [H(2)O(2)]/[triple bond MnO(2)] was 3.92, the rate constants (kMnO(2)) was 0.061 (min mM)(-1) as maximum. Oxygen production showed that the initial rates increased with decreasing [H(2)O(2)]/[triple bond MnO(2)] and the total amounts of oxygen was slightly less than the stoichiometric value (0.5) in most experiments. However, oxygen was produced at more than 0.5 in low [H(2)O(2)]/[triple bond MnO(2)] (i.e. 3.92 and 9.79). The relative production of hydroperoxide/superoxide anion implied that the production increased with low [H(2)O(2)]/[triple bond MnO(2)], and the existence of anions suggested that the mechanism includes propagation reactions with intermediates such as hydroperoxide/superoxide anion in solution. In addition, both [H(2)O(2)] decomposition and the production of anion were accelerated in alkaline solution. Manganese ion dissolved into solution was negligible in neutral and alkaline conditions, but it greatly increased in acidic conditions. PMID- 19136142 TI - Structural and physiological responses to ozone in Manna ash (Fraxinus ornus L.) leaves of seedlings and mature trees under controlled and ambient conditions. AB - Leaf-level microscopical symptom structure and physiological responses were investigated in seedlings experimentally exposed to ozone (O3) in indoor chambers (150 ppb, 8 h d(-1)/7 weeks), and field trees of Manna ash (Fraxinus ornus) exposed to ambient O3 (max 93 ppb/one growing season). Ozone-induced leaf injury, including leaf reddening and stippling, was observed in both seedlings and mature trees, but the morphology of injury in the stipples differed, being hypersensitive-like (HR-like) in the chamber seedlings and accelerated cell senescence (ACS) in the field trees. In both exposure conditions, the main structural impact of O3 was on the mesophyll and especially the upper assimilating cell layers. The main physiological impact was on carbon assimilation and on stomatal sluggishness. These effects were not due to stomatal structural injury and were more severe in juvenile compared to mature trees because of environmental (water availability, light) and constitutional (gas exchange capacity) factors and differences in the cell physiology processes (HR like vs. ACS) triggered by ozone stress. Given the plasticity of plant responses to ozone stress, dose/response relationships for tree seedlings in the indoor chambers cannot be extrapolated to mature trees unless ambient conditions are closely simulated. PMID- 19136144 TI - Sleep as a marker in the effective management of chronic osteoarthritis pain with opioid analgesics. AB - OBJECTIVES: Sleep disturbances frequently accompany chronic pain from osteoarthritis (OA), and their effective management may reflect successful treatment of chronic pain. The objective of this article is to provide a rationale for using improvement in sleep as a marker for effective management of chronic OA pain with opioid analgesics. For this purpose, available evidence evaluating the relationship between successful management of chronic pain with opioids and improvements in sleep in patients with OA is reviewed. METHODS: We conducted a comprehensive PubMed search to identify studies that systematically measured the impact of opioid treatment on pain and sleep parameters in the context of chronic pain from OA. Our search criteria included publication in a recognized peer-reviewed journal, randomized placebo-controlled design, and assessment of both pain intensity and sleep as outcomes. RESULTS: In each of the 10 placebo-controlled studies identified, concurrent improvements in pain intensity and measured sleep disturbances were observed in patients receiving the long-acting opioid analgesics under study. Improved overall sleep quality, reduced awakenings from pain, and increased duration of sleep were among the favorable changes observed in patients with OA treated with long-acting opioids. CONCLUSIONS: Current evidence suggests that various long-acting opioid analgesics simultaneously achieve pain control and improve sleep. However, the complex interaction between reduced pain and improved sleep requires further study. PMID- 19136145 TI - Does colorectal cancer clinical advancement affect adhesion molecules (sP selectin, sE-selectin and ICAM-1) concentration? AB - Adhesion molecules take part in physiological and pathological processes. They involved in inflammatory reactions and play important role in tumor invasion and the development of metastases. Soluble forms of P-selectin, E-selectin and ICAM-1 have been described in this study in patient with colorectal cancer. Plasma was obtained from 44 patients with colorectal cancer and 34 control subject's prior surgery, by an enzyme-linked immunosorbent assay (ELISA). The patients were divided according to TNM classification. Plasma level of all three molecules was significantly higher in colorectal cancer patients than in the control (p < 0.001). The highest level of sE-selectin and ICAM-1 were observed in patients with liver metastasis. There was no correlation between sP-selectin and sE selectin, but we found a significant correlation between sE-selectin and ICAM-1 in all patients. These findings suggest that plasma concentration of E-selectin and ICAM-1 may indicate tumor progression and liver metastasis. PMID- 19136143 TI - Understanding the epidemiology and progression of systemic lupus erythematosus. AB - OBJECTIVES: This review examines the burden and patterns of disease in systemic lupus erythematosus (SLE) and the influence and interactions of gender, ethnicity, age, and psychosocial attributes with respect to disease progression, focusing on issues relevant to clinical practice and research. METHODS: PubMed literature search complemented by review of bibliographies listed in identified articles. RESULTS: An increased risk among reproductive age women is clearly seen in African Americans in the United States. However, in other populations, a different pattern is generally seen, with the highest age-specific incidence rates occurring in women after age 40 years. The disease is 2 to 4 times more frequent, and more severe, among nonwhite populations around the world and tends to be more severe in men and in pediatric and late-onset lupus. SLE patients now experience a higher than 90% survival rate at 5 years. The less favorable survival experience of ethnic minorities is possibly related to socioeconomic status rather than to ethnicity per se, and adequate social support has been shown to be a protective factor, in general, in SLE patients. Discordance between physician and patient ratings of disease activity may affect quality of care. CONCLUSIONS: Our understanding of ways to improve outcomes in SLE patients could benefit from patient-oriented research focusing on many dimensions of disease burden. Promising research initiatives include the inclusion of community-based patients in longitudinal studies, use of self-assessment tools for rating disease damage and activity, and a focus on self-perceived disease activity and treatment compliance. PMID- 19136146 TI - Plasma homocysteine level and left ventricular thrombus formation in acute anterior myocardial infarction patients following thrombolytic therapy with t-PA. AB - AIMS: The aim of this study was to evaluate the relationship between homocysteine levels and the development of left ventricular thrombus in acute anterior myocardial infarction patients directed to thrombolytic therapy. METHODS AND RESULTS: Seventy-nine patients presenting with ST elevated acute anterior myocardial infarction and treated with thrombolytic agent, t-PA, were included in the study. Two-dimensional echocardiography was used to divide patients into 2 groups according to the presence (n = 14) or absence (n = 65) of thrombus in the left ventricle following myocardial infarction. The levels of fasting plasma total homocysteine, total cholesterol, triglycerides, HDL-cholesterol, LDL cholesterol, vitamin B12 and folic acid were assessed. There were no significant differences between two groups in terms of age, gender, hyperlipidemia and smoking. History of diabetes mellitus (28.57% versus 6.15%, p = 0.04), peak creatine phosphokinase levels (4153.54 +/- 1228.41 U/L versus 2456.92 +/- 1421.36 U/L, p < 0.001), mean left ventricular wall motion score index (2.21 +/- 0.18 versus 1.83 +/- 0.23, p < 0.001) and total fasting homocysteine levels (18.24 +/- 5.67 mmol/L versus 12.31 +/- 3.52 mmol/L, p < 0.001) were significantly higher in patients with left ventricular thrombus. In multivariate analysis; only diabetes mellitus (p = 0.03), higher wall motion score index (p = 0.001) and higher homocysteine levels (p = 0.04) were independent predictors of left ventricular thrombus formation. CONCLUSION: Our results suggest that; diabetes mellitus, higher wall motion score index and hyperhomocysteinemia independently increases the risk for the development of left ventricular thrombus formation in patients with acute anterior myocardial infarction following thrombolytic therapy. PMID- 19136147 TI - Bisphosphonate-induced osteonecrosis of the jaw (ONJ): Incidence and risk factors in patients with breast cancer and gynecological malignancies. AB - OBJECTIVE: Since 2003, multiple cases of bisphosphonate-induced osteonecrosis of the jaw (ONJ) were reported. The aim of this study was to describe the incidence and risk factors of ONJ in patients with breast cancer or gynecological malignancies receiving bisphosphonates (BP). METHODS: ONJ was recorded for all patients with breast cancer or gynecological malignancies treated with intravenous bisphosphonates at the Department of Gynecology and Obstetrics, University Hospital Tuebingen during April, 1999 and May, 2006. RESULTS: 10 of 345 (2.9%) patients with breast cancer or gynecological malignancies developed ONJ while receiving bisphosphonate therapy. Six patients with ONJ had a history of recent dental procedures. All patients had received zoledronic acid as part of their bisphosphonate regimen. Time of exposure to bisphosphonates and the number of treatment cycles were significant risk factors for the development of ONJ (p<0.001). In patients diagnosed with ONJ the mean number of treatment cycles was 27+/-18 cycles. However, the mean number of treatment cycles in patients without manifestation of ONJ was 12+/-12 cycles. CONCLUSION: Length of exposure to BPs and the cumulative dose of given BPs seem to be the most important risk factors for the development of ONJ followed by dental procedures. PMID- 19136148 TI - Specialized and high-volume care leads to better outcomes of ovarian cancer treatment in the Netherlands. AB - OBJECTIVE: We investigated the influence of hospital and gynecologist level of specialization and volume on surgical results and on survival of ovarian cancer patients. METHODS: Data were collected from 1077 ovarian cancer patients treated from 1996 to 2003 in a random sample of 18 Dutch hospitals. Hospitals and gynecologists were classified according to specialization (general, semi specialized or specialized) and by volume (12 cases/year). Outcomes were percentage of adequately staged and optimally debulked patients and length of overall survival. Data were analyzed using multivariable logistic regression (surgical results) and Cox regression (survival). RESULTS: The level of specialization and the volume of hospitals and of gynecologists were strongly related to the proportion of adequately staged patients (adjusted odds ratio (OR) specialized hospitals 3.9 (95% confidence interval (CI) 2.0-7.6); specialized gynecologists 9.5 (95% CI 4.7-19)). Patients with stage III disease had a higher chance of optimal debulking when treated in specialized hospitals (adjusted OR 1.7 (95% CI 1.1-2.7)) or by high volume gynecologists (adjusted OR 2.8 (95% CI 1.4-5.7)). Overall survival was best in patients treated in specialized hospitals and by high-volume gynecologists. CONCLUSION: The specialization level of hospitals and the surgical volume of gynecologists positively influence outcomes of surgery and survival. Concentration of ovarian cancer care thus seems warranted. PMID- 19136149 TI - Flowability of composites is no guarantee for contraction stress reduction. AB - OBJECTIVES: The purpose of this study was to measure the contraction stress development of three flowable resin-composite materials (Grandio Flow, VOCO GmbH, Cuxhaven, Germany; Tetric Flow, Ivoclar Vivadent, Schaan, Liechtenstein; Filtek Supreme XT Flowable Restorative, 3M ESPE, ST. Paul, MN, USA) and an universal micro-hybrid composite resin (Filtek Z250, 3M ESPE, St. Paul, MN, USA) during photopolymerization with a halogen curing light, using a novel stress-measuring gauge. METHODS: Curing shrinkage stress was measured using a stress-analyzer. Composites were polymerized with a halogen curing unit (VIP, Bisco Inc., Schaumburg, IL, USA) for 40s. The contraction force (N) generated during polymerization was continuously recorded for 180s after photo-initiation. Contraction stress (MPa) was calculated at 20s, 40s, 60s, 120s and 180s. Data were statistically analyzed. RESULTS: Filtek Supreme XT Flowable Restorative exhibited the highest stress values compared to other materials (p<0.05), while the lowest values were recorded with Tetric Flow (p<0.05). Tetric Flow was also the only flowable composite showing stress values lower than the conventional composite Filtek Z250 (p<0.05). SIGNIFICANCE: Flowable composites investigated with this experimental setup showed shrinkage stress comparable to conventional resin restorative materials, thus supporting the hypothesis that the use of flowable materials do not lead to marked stress reduction and the risk of debonding at the adhesive interface as a result of polymerization contraction is similar for both type of materials. PMID- 19136150 TI - Long-term clinical outcomes following the use of synthetic hydroxyapatite and bone graft in impaction in revision hip arthroplasty. AB - Impaction grafting using morsellised allograft bone restores bone stock, but carries the potential for transmission of infection. Synthetic bone graft substitutes can eliminate this risk but may, however, influence outcome. In this study we tested the hypothesis that a 50/50 mix of hydroxyapatite and allograft does not affect long-term function, survival or radiological outcome. Sixty-five patients had revision hip arthroplasty using impaction grafting with either pure allograft (42 patients) or a 50/50 mixture of allograft and solid particulate hydroxyapatite. Harris hip scores were assessed pre-operatively and annual intervals thereafter. Function was analyzed using multilevel modeling, the Kaplan Meier method used for survival analysis and graft incorporation was assessed radiologically. The hip score improved in both groups but showed a small annual decline (average 1.2/year, p<0.01). This decline was higher for females (average 3.4, p=0.025) and significantly related to pre-op scores (p<0.001). After adjusting for these, allograft patients had marginally higher scores (difference=3.1, p=0.3). The majority of revisions were for aseptic loosening. At 13 years survival in the allograft group was 84%, and 82% in the mixture group (p=0.96, log rank test). Radiologically the graft incorporation was similar in both groups (p=0.62). We conclude that long-term prosthesis survival and function following revision arthroplasty with a 50/50 mixture of allograft and hydroxyapatite are comparable to allograft alone. PMID- 19136151 TI - Direct cell entry of gold/iron-oxide magnetic nanoparticles in adenovirus mediated gene delivery. AB - Gold/iron-oxide MAgnetic Nanoparticles (GoldMAN) imparts useful magnetic properties to various biomolecules. Gold nanoparticles immobilized on the surface of magnetic nanoparticles allow for the conjugation of biomolecules via an Au-S bond. Here, we present a practical application by utilizing GoldMAN and a magnetic field to induce intracellular transduction. This method has great potential for application of the adenovirus gene delivery vector (Ad), widely used for in vitro/in vivo gene transfer, to Ad-resistant cells. We demonstrated that Ad was easily immobilized on GoldMAN and the Ad/GoldMAN complex was introduced into the cell by the magnetic field, which increased gene expression over 1000 times that of Ad alone. The GoldMAN penetrated the plasma membrane directly, independent of the cell-surface virus receptors and endocytosis pathway. This mechanism will contribute to improve the gene expression efficiency of Ad. This technology is a useful tool for extending Ad tropism and enhancing transduction efficiency. GoldMAN also makes possible the effective use of various biomolecules within the cell because of its interesting cell-entry mechanism. PMID- 19136152 TI - Reconstruction of calvarial defect using a tricalcium phosphate-oligomeric proanthocyanidins cross-linked gelatin composite. AB - A biodegradable GTP composite which was composed of oligomeric proanthocyanidins (OPCs) cross-linked gelatin mixed with tricalcium phosphate was developed as a bone substitute. The subcutaneous implantation in rats was examined to determine the in vivo degradation and biocompatibility of the GTP composites with various cross-linking densities. Experimental results indicated that the rate of in vivo degradation was markedly attenuated as the concentration of OPCs increased above 5.0wt%. Furthermore, this study examined the biological response of rabbit calvarial bone to GTP composite to evaluate its potential for use as an osteoconductive bone substitute. Bone defects (10mm in diameter) in New Zealand white rabbits were filled with the GTP composite. The de-protenized bovine cancellous bone matrix was employed as the control material. The results of radiographic analyses demonstrated obviously greater new bone ingrowth in the GTP composite than in the de-proteinized bovine bone at the same implantation time. Progressive replacement of the GTP composite by new bone proceeded by a combination of osteoconduction and biodegradation. The biodegradable GTP composite thus has great potential for improving bone repair. PMID- 19136154 TI - Social problem-solving in high-functioning schizophrenia: specific deficits in sending skills. AB - This study examined social problem-solving performance in high-functioning schizophrenia (n=26) and its relation to neurocognition. Ten healthy controls were used as a comparison group. Social problem-solving was assessed with the Assessment of Interpersonal Problem Solving Skills (AIPSS) method. The schizophrenia group was outperformed by healthy controls on all AIPSS measures, reaching statistical significance for sending skills. Exploration of the internal relationship between different aspects of social problem-solving showed that identification of an interpersonal problem (a receiving skill) was not correlated with formulating solutions to the problem (processing skills) or successfully role-playing solutions (interpersonal sending skills). Non-verbal performance in the role-play (an interpersonal sending skill) was not significantly correlated with identification of an interpersonal problem or the generation of solutions. This suggests a dissociation of social problem-solving processes. Social problem solving was significantly associated with psychomotor speed, verbal learning, semantic fluency and cognitive flexibility. Clinical implications are that remediation of social problem-solving skills should focus on role-playing (nonverbal) interpersonal behaviors, rather than on verbally analyzing an interpersonal problem and clarifying alternative solutions. PMID- 19136153 TI - The influence of BCR density on the differentiation of natural poly-reactive B cells begins at an early stage of B cell development. AB - B cell antigen receptor (BCR) density plays a role in the differentiation of immature B cells to their mature compartments; however, the exact strategy of its influence on the development of natural autoreactive B cells is still unclear. In the present study, we explored the role of BCR surface density in autoreactive B cell development by studying two lines of mice containing distinct copy numbers of an IgH transgene with V(H) derived from a poly-reactive natural antibody 3B4. Surface BCR levels were found to be related to the transgene copy number in these mice. In mice with higher copy numbers of the transgene, the BCRs were found to promote the remaining of autoreactive B cells into marginal zone (MZ) and B-1a subsets; meanwhile, elevated surface BCR levels were correlated with a significant decrease of follicular (Fo) B cell numbers and a reduction in the number of multiple stages of immature B cells both in spleen and bone marrow (BM). Interestingly, no difference in the ratio of cell apoptosis and proliferation was found in all stages of B cell development between two lines, except that more severely aberrant proliferation of pro/pre-B cells in BM was found in mice with higher transgene copies. This data supports the idea that natural poly-reactive B cells can be positively selected into MZ and B-1 cells, and high BCR surface density favors this selection. More importantly, our data suggests that the influence by receptor expression on the differentiation of natural poly-reactive B cells begins at an early stage of B cell development. PMID- 19136155 TI - Characterization and expression profile of complete functional domain of granulysin/NK-lysin homologue (buffalo-lysin) gene of water buffalo (Bubalus bubalis). AB - Granulysin (GNLY)/NK-lysin (NKL) is an effector antimicrobial cationic peptide expressed in the cytotoxic and natural killer lymphocytes. We report here cDNA sequence (405bp) encoding the complete functional domain of buffalo-lysin (bu lysin), and its expression profile in the various tissues. The nucleotide sequence of bu-lysin exhibited >85% identity with the bovine lysin. Comparison of the deduced amino acid sequence of bu-lysin with those of GNLY/NKL of different species revealed the conservation of six cysteine (Cys) residues and five alpha helices. Unlike the homologues in other species, bu-lysin composed of 11 positively charged Lys residues as in equine. The expression of bu-lysin mRNA in the in vitro cultured lymphocytes was inducible and increased markedly (p<0.05) in a dose dependant manner when incubated with Concanavalin A (ConA). The expression of bu-lysin mRNA in the different tissues was variable: comparatively higher in the spleen and lymph node, moderate in the uterine endometrium and low in the liver and kidney. These results indicate the existence and active expression of GNLY/NKL homologue in water buffalo having a significant influence in immune response. PMID- 19136156 TI - The effect of lidocaine on in vitro adhesion and migration of equine neutrophils. AB - The effect of lidocaine on in vitro migration and adhesion of equine neutrophils was evaluated. Neutrophils were isolated from equine whole blood using a Percoll gradient centrifugation protocol. Purified neutrophils were incubated with lidocaine at concentrations from 0.1 to 1000 microg/ml for 30 min at 37 degrees C, after calcein loading. Neutrophil integrin-mediated adhesion in response to stimulation with 100 nM LTB(4), 100 nM PAF, or 100 ng/ml IL-8, or integrin mediated migration in response to stimulation with 100 nM LTB(4), 150 nM PAF, or 100 ng/ml IL-8 was assessed. Statistical significance was set at P<0.05. Neutrophil adhesion was significantly increased in response to all three stimulants. IL-8-stimulated adhesion was significantly increased when neutrophils were incubated with 1mg/ml lidocaine, compared to lower lidocaine concentrations. LTB(4)-stimulated adhesion was significantly increased when neutrophils were incubated with 1mg/ml lidocaine compared to that at 5 microg/ml lidocaine. Migration was significantly increased in response to IL-8. IL-8 and LTB(4) stimulated migration was significantly increased when neutrophils were incubated with 1mg/ml lidocaine, compared to lower lidocaine concentrations. In conclusion, lidocaine did not inhibit neutrophil migration or adhesion in vitro at therapeutic concentrations, and increased migration and adhesion at higher concentrations. PMID- 19136157 TI - Screening to detect permanent childhood hearing impairment in neonates transferred from the newborn nursery. AB - OBJECTIVES: The focus of this report is hearing screening of newborns transferred from the regular nursery to a specialized area. The purpose of the study undertaken was: (1) to determine whether screening coverage in this population was achieved; (2) to establish whether the linkage between neonatal screening and the diagnostic follow-up was carried out correctly; (3) to better determine the incidence of permanent childhood hearing impairment (PCHI) in this at-risk population. METHODS: Six population centres averaging 12,000 births annually participated (Bordeaux, Lille, Paris, Marseille, Toulouse and Lyon). Automated auditory brainstem response (AABR) (Natus ALGO 3i) screening was performed in two stages: i.e. infants with initial "positive" results were screened a second time using the same technique. Of the 117,103 babies born during the study period, 4972 neonates were "transferred" and comprised the population for this report (4.2% of the total births). RESULTS AND DISCUSSION: Screening results for 4972 "transferred" neonates were compared with those of non-transferred neonates (N=112,131). Screening coverage of eligible infants was significantly lower (75.4%) in "transferred" neonates (3750 infants screened) compared to 97.5% coverage of non-transferred neonates (109,349 infants screened). The rate of positive results after the first stage AABR was higher in the "transferred" population (11.1%) than in the non-transferred population (6.5%). Of the 415 "transferred" newborns with initial positive screens, 91.3% were rechecked as stipulated in the project protocol. The second pre-discharge AABR ascertained that in half of the cases auditory function had normalized in the day. Of the 183 "transferred" infants whose result remained suspect at the conclusion of both stages of the neonatal screen (4.9% of the tested population), only 70.5% returned to the audiology centre for diagnostic follow-up. The incidence of bilateral PCHI was markedly higher (4/1000) in "transferred" infants than in the non-transferred population (1.08/1000). CONCLUSIONS: The difficulty of obtaining universal screening coverage in "transferred" infants was, unfortunately, verified in this prospective, multicentre study. Further, the diversity of our "transferred" population was not much greater than that revealed by careful analysis of published hearing screening studies in neonatal intensive care unit (NICU) infants. The influence of risk factors and their more or less complex combinations is apparent. PMID- 19136158 TI - Temporal maps and informativeness in associative learning. AB - Neurobiological research on learning assumes that temporal contiguity is essential for association formation, but what constitutes temporal contiguity has never been specified. We review evidence that learning depends, instead, on learning a temporal map. Temporal relations between events are encoded even from single experiences. The speed with which an anticipatory response emerges is proportional to the informativeness of the encoded relation between a predictive stimulus or event and the event it predicts. This principle yields a quantitative account of the heretofore undefined, but theoretically crucial, concept of temporal pairing, an account in quantitative accord with surprising experimental findings. The same principle explains the basic results in the cue competition literature, which motivated the Rescorla-Wagner model and most other contemporary models of associative learning. The essential feature of a memory mechanism in this account is its ability to encode quantitative information. PMID- 19136159 TI - Relationship between clinical phenotypes and cognitive impairment in Parkinson's disease (PD). AB - Most patients with idiopathic PD (IPD) show variable degrees of cognitive decline. The purpose of this study was to evaluate the relationship between the predominant motor symptom at the time of disease onset and the level of cognitive function in patients with IPD. A total of 159 patients with IPD were enrolled in this study. The patients' initial motor symptoms were classified into three types: tremor-dominant (TD), bradykinesia and rigidity-dominant (BRD), gait and postural instability-dominant (GPD). Disease severity was rated according to the Hoehn-Yahr classification (H&Y stage). Overall cognitive status was evaluated using the Korean versions of the Mini-Mental State Examination (K-MMSE) and the Modified Mini-Mental State (3MS) tests. The GPD group showed the lowest scores of the K-MMSE/3MS, and the patients with TD showed the best performance in the cognitive analysis (p<0.05). The patients who were older at disease onset showed worse cognitive performance than those the patients who were younger at disease onset (p<0.05). There was no difference in cognitive status according to H&Y stages. The accurate classification of initial motor symptoms and the detailed history, including the exact onset age of IPD, may allow us to predict cognitive decline in IPD. PMID- 19136160 TI - The effect of trust and change in trust on self-rated health: a longitudinal study among aging people. AB - This study examined whether trust predicted subsequent self-rated health over time at 3 years follow-up among aging people, and whether changes in trust were associated with self-rated health. Longitudinal, questionnaire-based data were collected from three age cohorts (born in 1926-1930, 1936-1940, and 1946-1950) living in the Province of Paijat-Hame, southern Finland. The response rate at the baseline in 2002 was 66% (n=2815). The follow-up was carried out in 2005, with 79% of eligible individuals participating (n=2216). Logistic regression analyses were used to derive the results. High trust was a strong predictor for good self rated health at the follow-up. Adjusting for background variables, however, attenuated the association. In addition, good self-rated health was most common among men with sustained high trust, among women the association was somewhat weaker. Among men improvement in trust was associated with good self-rated health, but this correlation weakened after multiple adjustments. Thus, longitudinally trust is an important contributor to self-rated health among aging people. Moreover, improvement of trust but also the stability of high trust especially among men indicate better self-rated health. Trust has a positive effect on health and should therefore be seen as a significant element in health promotion. PMID- 19136161 TI - Comparison and correlates of participation in older adults without disabilities. AB - This study was conducted to compare, by age group and gender, the level of participation of older adults who had no disabilities, and to determine which characteristics are most associated with participation. This study involved 350 randomly recruited community-dwelling older adults. Participation in daily activities and social roles were measured with the Assessment of Life Habits (LIFE-H). Demographic, health-related and environmental data were also collected. A decline with age was observed in four of the six daily activities domains and two of the four social roles domains of participation. However, these lower scores are mainly explained by the 85+ group, which consistently scored lower than the 65-69-year-old group. No differences were found between the 65-69, 70-74 and 75-79 years old groups. Some participation domains differed according to gender. Satisfaction with participation was high and did not differ between age groups. Characteristics most associated with participation vary according to the domains; generally, age and marital status are the best determinants of participation. This study found that most of older adults have an unrestricted level of participation which decreases only late in the aging process. This reduction in participation in very old adults was not accompanied by a decrease in satisfaction, supporting the hypothesis that they can participate satisfactorily in valued activities. PMID- 19136162 TI - QRS prolongation during exercise stress testing: beyond ST segment depression. PMID- 19136163 TI - Gross echocardiographic underestimation of mitral valve calcification--a near fatal experience. AB - Acute severe mitral regurgitation is a feared complication following percutaneous balloon mitral valvuloplasty (PBMV) for the treatment of severe mitral stenosis, contributing to the 1% peri-procedural mortality rate of this procedure. Whilst there is an extensive experience with the use of the Wilkins echocardiographic score to assess patient suitability and predict long-term clinical outcomes following PBMV, catastrophic severe acute mitral regurgitation, such as that described in our patient, is a remarkably rare occurrence following PBMV in patients with favourable Wilkins echocardiographic criteria. We highlight a case of the gross underestimation of the degree of valvular calcification using trans oesophageal echocardiography, when compared to the findings at surgery, which contributed to our patients' life-threatening mitral regurgitation following the first balloon inflation. We advocate further research into the utility of multi detector computed tomography (MDCT) imaging for the adjunctive pre-procedural assessment of the degree mitral calcification in order to further minimise the risk of peri-procedural complications associated with PBMV. This would be particularly suitable in the elderly population who normally have greater degrees of valvular calcification that may be underestimated with echocardiography alone. PMID- 19136164 TI - Pre-injection of magnesium sulfate enhances the efficacy of ibutilide for the conversion of typical but not of atypical persistent atrial flutter. AB - BACKGROUND: Ibutilide is a class III antiarrhythmic drug, frequently used for conversion of atrial fibrillation and flutter. Retrospective cohort evaluations found that intravenous application of magnesium enhances the efficacy of ibutilide for chemical conversion of these arrhythmias. This prospective study sought to investigate the effects of intravenously pre-injected magnesium on the conversion rate of ibutilide for typical and atypical atrial flutter. METHODS: We performed a prospective, randomized, placebo-controlled study. Patients with typical atrial flutter (TAF) or atypical atrial flutter (AAF) were randomized to receive either 4 g of intravenous magnesium sulfate or placebo immediately before administration of a maximum dose of 2 mg of ibutilide fumarate. Continuous rhythm monitoring for 4 h provided information on conversion to sinus rhythm. QT interval durations were measured before randomization, after magnesium, as well as 30 min and 4 h after starting ibutilide infusion. RESULTS: We randomized 117 patients (58 with and 59 without pre-injection of magnesium; 65 with TAF and 52 with AAF). In patients with TAF, pre-injection of magnesium significantly improved the efficacy of ibutilide for conversion (85% with magnesium vs. 59% with placebo, p=0.017). In patients with AAF, no significant difference in conversion rates between patients receiving magnesium or placebo was detected (48% vs. 56%, p=0.189). Pre-injection of magnesium did not significantly influence the QT intervals at any time after administration of ibutilide. CONCLUSIONS: Pre-injection of magnesium significantly enhances the efficacy of ibutilide for the conversion of TAF but not of AAF. PMID- 19136165 TI - Late thrombotic occlusion of a left internal mammary artery graft causing ST elevation myocardial infarction. AB - Late occlusion of the left internal mammary artery (LIMA) is rare, and its pathological basis is unknown. We present a case of acute occlusion of the LIMA in a 77-year-old woman, 7 years after graft surgery, causing fatal myocardial infarction. Diagnostic angiography of the LIMA revealed a proximal occlusion just beyond the LIMA origin which re-canalised after repeated injection of contrast with near complete restoration of LIMA patency and flow with smooth angiographic appearance. Despite initial diminution of pain and electrocardiographic changes with recanalisation, pain and ST changes returned and the patient died. Restoration of near normal vessel patency with angiography suggests that the basis of this occlusion is not atherosclerotic plaque rupture, but is more likely to represent in situ thrombosis or, less likely, localized spasm. Early recognition of the possibility of LIMA occlusion may direct initial angiography to the LIMA rather than the native circulation, and reduce the time to reperfusion. PMID- 19136166 TI - Mirror-image type D interrupted aortic arch: a novel cardiac phenotype providing some perspective in the del22q11.2 syndrome. AB - A 1-day-old baby boy was referred from a local obstetric clinic due to feeble crying, tachypnea, and tachycardia. Two-dimensional echocardiography with Doppler and multidetector computer tomography showed mirror-image type D interrupted aortic arch, conotruncal ventricular septal defect, and patent ductus arteriosus. Emergency cardiac surgery by a biventricular Norwood procedure was performed to relieve his symptom and sign of congestive heart failure successfully. Thymus could not be seen by surgical exploration of the superior mediastinum. Serum C terminal parathyroid hormone was decreased to less than the normal lower limit. Cytogenetic analysis and fluorescence in situ hybridization study of blood revealed a deletion in chromosome 22q11.2. To the best of our knowledge, mirror image type D interrupted aortic arch has never been reported in patients with the del22q11.2 syndrome in the English literature. This unusual aortic arch anomaly may provide us a new perspective in the spectrum of cardiovascular malformations in the del22q11.2 syndrome and advocate 22q11.2 deletion as one of the genetic causes of some rare aortic arch anomalies and their correspondent mirror-images. PMID- 19136167 TI - Factors of importance for patients' decision time in acute coronary syndrome. AB - BACKGROUND: Not much is known about the patients' decision time in acute coronary syndrome (ACS). The aim of the survey was therefore to describe patients' decision time and factors associated with this parameter in ACS. METHODS: We conducted a national survey comprising intensive cardiac care units at 11 hospitals in Sweden in which patients with ACS diagnosis and symptoms onset outside hospital participated. Main outcome measures were patients' decision time and factors associated with patients' decision time. RESULTS: In all, 1939 patients took part in the survey. The major factors associated with a shorter patient decision time were: 1) ST-elevation ACS, 2) associated symptoms such as vertigo or near syncope, 3) interpreting the symptoms as cardiac in origin, 4) pain appearing suddenly and reaching a maximum within minutes, 5) having knowledge of the importance of quickly seeking medical care and 6) experiencing the symptoms as frightening. The following aspects of the disease were associated with a longer decision time: 1) pain was localised in the back and 2) symptom onset at home when alone. CONCLUSION: A number of factors, including the type of ACS, the type and localisation of symptoms, the place where symptoms occurred, patients' interpretation of symptoms and knowledge were all associated with patients' decision time in connection with ACS. PMID- 19136168 TI - Targeting converging therapeutic pathways to overcome hypertension. PMID- 19136169 TI - Novel mutation (H402R) in the S1 domain of KCNH2-encoded gene associated with long QT syndrome in a Spanish family. AB - Long-QT syndrome is a congenital cardiac disease resulting in ventricular arrhythmias and sudden death. Genetic mutations in two protein ion-channel genes, KCNQ1 and KCNH2. The mutations position in these genes provides additional information about the evaluation of the risk-stratification. In a Spanish family in whom previous repetitive syncope episodes, sudden death and pathological prolongation of the QT interval were documented, a novel heterozygous mutation in the KCNH2 gene (A1218>G) was identified. This mutation loading to amino acid substitution H420R in the S1 transmembrane domain of KCNH2. The new A1218>G mutation in the KCNH2 gene detected in this Spanish family causes arrhythmia manifestation in the carriers. PMID- 19136170 TI - WITHDRAWN: Resynchronization of the infarcted myocardium. AB - This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19136171 TI - Advanced algorithms can lead to electrocardiographic misinterpretations. AB - We observed a patient with syncope, who implanted a pacemaker with advanced algorithms such as "atrial-tachy response" and "dynamic atrio-ventricular delay". After one year, conventional ECG Holter showed pacemaker malfunction, wrongly attributed to exposure to electromagnetic field. In fact, telemetry revealed an inappropriate programming and solved our case. Holter monitoring is commonly performed in the evaluation of pacemaker malfunction, albeit it remains a quite shallow diagnostic method especially to detect electromagnetic interferences. New algorithms seem important, but it is reasonable to obtain more suitable analytical tools, too. PMID- 19136172 TI - A combination of right ventricular hypertrabeculation/noncompaction and Ebstein's anomaly. AB - A 35-year-old man admitted with iterative extremities edema for three months and mild chest complaint associated with activity without chest distress for five days. He underwent an initial 2DE that did not confirm HVM/NVM and 3DE that strengthened the diagnosis of HVM/NVM. The limitations of 2DE may lead to misinterpretation of prominent trabeculations as "false tendons," and may also lead to underestimation of the severity of HVM/NVM. In contrast to 2DE, 3DE provides for pyramid-shaped datasets that encompass the entire right ventricular. Specifically, right ventricular can be sectioned in userselected planes and an unlimited number and angles of such planes can be used. Intracavitary echodensities that are suspicious for trabeculations can be tracked in multiple directions from base to apex. To our knowledge, this is the first report on only right ventricular HVM/NVM associated with Ebstein's malformation. The leaflets of the tricuspid valve develop from endocardial cushion tissue and the myocardium. The tricuspid valve leaflets become freely movable after delamination of the tension apparatus within the myocardium. Although the coexistence of right ventricular HVM/NVM and Ebstein's malformation could be a coincidence, I believe that both defects could be caused by a developmental arrest of the right ventricular myocardium. That is to say, the genetic change that is responsible for Ebstein's anomaly also plays a role in the differentiation of the right myocardium. PMID- 19136173 TI - Stress-induced hypercontractility in patients with hypertension: an interesting imaging finding. AB - Excessive sympathetic activity and stress-induced LV hypercontractility may be detected in hypertensives or hypertensive LV hypertrophy. Recent quantitative data support that hypertensive patients may be associated with stress-induced LV hypercontractility. Stress-induced hypercontractility may also be related to basal LV cavity obliteration and dynamic LV outflow tract obstruction in hypertensives. In contrast to LV cavity dilation, stress-induced LV cavity obliteration may be associated with favorable outcome, however it has not been completely elucidated. Hypertensives with dynamic LVOT obstruction may clinically present with acute heart failure and normal systolic function. LV contractility may change and evaluation of LV geometry, contractility and volume by imaging techniques may be important in disease progression. PMID- 19136174 TI - Enhanced proliferation of acinar and progenitor cells by prophylactic pilocarpine treatment underlies the observed amelioration of radiation injury to parotid glands. AB - BACKGROUND: Administration of pilocarpine before irradiation can ameliorate radiation-induced hyposalivation. Indirect evidence suggests that this effect may be mediated through induction of a compensatory response. In this study, this hypothesis is tested directly, by assessing the proliferation of progenitor and secretory cells in irradiated and non-irradiated parotid gland tissue. METHODS: In a rat model, parotid glands were unilaterally irradiated with a single dose of 15 Gy, 60 min after administration of pilocarpine (4.0mg/kg). Rats were sacrificed for proliferating cell nuclear antigen (PCNA) labelling, assessing the number of proliferating progenitor and secretory cells, before, and 10h, 1, 3, 7, 10, 20 and 30 days after irradiation. RESULTS: A small radiation-induced increase in PCNA expressing cells was observed, both in the acinar (secretory cells) and intercalated duct cell (containing the progenitor cells) compartment. This increment was significantly enhanced in pilocarpine pre-treated glands. In fact, in this group of animals increased proliferation was observed both in the irradiated and the shielded gland. CONCLUSIONS: Amelioration of early loss of rat salivary gland function after radiation by pilocarpine pre-treatment is, at least in part, due to compensatory mechanisms through increased proliferation of undamaged cells. PMID- 19136175 TI - Prospective evaluation of quality of life after permanent prostate brachytherapy with I-125: importance of baseline symptoms and of prostate-V150. AB - BACKGROUND AND PURPOSE: Detailed knowledge of quality of life (QoL) after permanent I-125 brachytherapy may aid in counselling patients with early-stage prostate cancer. MATERIALS AND METHODS: Seventy-four consecutive patients with low-risk prostate cancer were asked to complete the EORTC QLQ-C30 questionnaire with the prostate-specific PR25 module before implant, four weeks and one year after implant (response rates 97%, 88% and 89%, respectively). Implant characteristics were correlated with QoL scores. RESULTS: Global QoL was stable from pre-treatment to one year after implant and similar to age-adjusted scores of healthy controls. Significant changes versus baseline in QLQ-C30 domains were worsened social function at four weeks, increased constipation at four weeks and at one year and improved emotional function at one year. PR25 urinary symptoms were significantly increased at four weeks and, despite some improvement, at one year; bowel symptoms were slightly increased. Both types of symptoms were most strongly related with pre-treatment symptom scores. Prostate-V150 was the only implant parameter significantly associated with both urinary and bowel symptoms at four weeks and one year. CONCLUSIONS: Limiting the high-dose subvolume in the prostate may be beneficial to reduce urinary and bowel symptoms but the major determinant of symptoms after I-125 implant is the baseline symptom level. PMID- 19136177 TI - Yeast population dynamics during pilot-scale storage of grape marcs for the production of Grappa, a traditional Italian alcoholic beverage. AB - The composition and population dynamics of the yeast microflora of grape marcs were investigated during a pilot scale fermentation study using two white grape varieties, namely Moscato and Prosecco, from two distinct areas of the Veneto Region. Yeast counts were made at the beginning, after 4 and after 15 days of marc storage under anaerobic conditions. Seventy isolates from each sampling time were identified to species by RAPD-PCR analysis and subsequent ITS region sequencing. A good biodiversity of yeasts occurred in both marcs at the beginning of fermentation, with high presence of Hanseniaspora opuntiae, but without detectable presence of Saccharomyces strains, which instead became the dominant yeast after just 4 days of fermentation, remaining at that level until the end of fermentation. Colonization of Moscato marc by S. cerevisiae resulted better, in relation to its higher sugar content. Characterization of S. cerevisiae isolates by mitochondrial DNA restriction analysis revealed the presence of 66 different strains in the marc from the Moscato grapes, without the occurrence of a clearly dominant strain, while in the marc from the Prosecco grapes only 23 different profiles were scored, with a dominant strain that accounted for 62.7% of the Saccharomyces population after 4 days of fermentation. PMID- 19136176 TI - A comparison of risk assessments on Campylobacter in broiler meat. AB - In recent years, several quantitative risk assessments for Campylobacter in broiler meat have been developed to support risk managers in controlling this pathogen. The models encompass some or all of the consecutive stages in the broiler meat production chain: primary production, industrial processing, consumer food preparation, and the dose-response relationship. The modelling approaches vary between the models, and this has supported the progress of risk assessment as a research discipline. The risk assessments are not only used to assess the human incidence of campylobacteriosis due to contaminated broiler meat, but more importantly for analyses of the effects of control measures at different stages in the broiler meat production chain. This review paper provides a comparative overview of models developed in the United Kingdom, Denmark, the Netherlands and Germany, and aims to identify differences and similarities of these existing models. Risk assessments developed for FAO/WHO and in New Zealand are also briefly discussed. Although the dynamics of the existing models may differ substantially, there are some similar conclusions shared between all models. The continuous introduction of Campylobacter in flocks implies that monitoring for Campylobacter at the farm up to one week before slaughter may result in flocks that are falsely tested negative: once Campylobacter is established at the farm, the within-flock prevalence increases dramatically within a week. Consequently, at the point of slaughter, the prevalence is most likely to be either very low (<5%) or very high (>95%). In evaluating control strategies, all models find a negligible effect of logistic slaughter, the separate processing of positive and negative flocks. Also, all risk assessments conclude that the most effective intervention measures aim at reducing the Campylobacter concentration, rather than reducing the prevalence. During the stage where the consumer handles the food, cross-contamination is generally considered to be more relevant than undercooking. An important finding, shared by all, is that the tails of the distributions describing the variability in Campylobacter concentrations between meat products and meals determine the risks, not the mean values of those distributions. Although a unified model for risk assessment of Campylobacter in the broiler meat production would be desirable in order to promote a European harmonized approach, it is neither feasible nor desirable to merge the different models into one generic risk assessment model. The purpose of such a generic model has yet to be defined at a European level and the large variety in practices between countries, especially related to consumer food preparation and consumption, complicates a unified approach. PMID- 19136178 TI - Biogeochemistry at a wetland sediment-alluvial aquifer interface in a landfill leachate plume. AB - The biogeochemistry at the interface between sediments in a seasonally ponded wetland (slough) and an alluvial aquifer contaminated with landfill leachate was investigated to evaluate factors that can effect natural attenuation of landfill leachate contaminants in areas of groundwater/surface-water interaction. The biogeochemistry at the wetland-alluvial aquifer interface differed greatly between dry and wet conditions. During dry conditions (low water table), vertically upward discharge was focused at the center of the slough from the fringe of a landfill-derived ammonium plume in the underlying aquifer, resulting in transport of relatively low concentrations of ammonium to the slough sediments with dilution and dispersion as the primary attenuation mechanism. In contrast, during wet conditions (high water table), leachate-contaminated groundwater discharged upward near the upgradient slough bank, where ammonium concentrations in the aquifer where high. Relatively high concentrations of ammonium and other leachate constituents also were transported laterally through the slough porewater to the downgradient bank in wet conditions. Concentrations of the leachate-associated constituents chloride, ammonium, non-volatile dissolved organic carbon, alkalinity, and ferrous iron more than doubled in the slough porewater on the upgradient bank during wet conditions. Chloride, non-volatile dissolved organic carbon (DOC), and bicarbonate acted conservatively during lateral transport in the aquifer and slough porewater, whereas ammonium and potassium were strongly attenuated. Nitrogen isotope variations in ammonium and the distribution of ammonium compared to other cations indicated that sorption was the primary attenuation mechanism for ammonium during lateral transport in the aquifer and the slough porewater. Ammonium attenuation was less efficient, however, in the slough porewater than in the aquifer and possibly occurred by a different sorption mechanism. A stoichiometrically balanced increase in magnesium concentration with decreasing ammonium and potassium concentrations indicated that cation exchange was the sorption mechanism in the slough porewater. Only a partial mass balance could be determined for cations exchanged for ammonium and potassium in the aquifer, indicating that some irreversible sorption may be occurring. Although wetlands commonly are expected to decrease fluxes of contaminants in riparian environments, enhanced attenuation of the leachate contaminants in the slough sediment porewater compared to the aquifer was not observed in this study. The lack of enhanced attenuation can be attributed to the fact that the anoxic plume, comprised largely of recalcitrant DOC and reduced inorganic constituents, interacted with anoxic slough sediments and porewaters, rather than encountering a change in redox conditions that could cause transformation reactions. Nevertheless, the attenuation processes in the narrow zone of groundwater/surface-water interaction were effective in reducing ammonium concentrations by a factor of about 3 during lateral transport across the slough and by a factor of 2 to 10 before release to the surface water. Slough porewater geochemistry also indicated that the slough could be a source of sulfate in dry conditions, potentially providing a terminal electron acceptor for natural attenuation of organic compounds in the leachate plume. PMID- 19136179 TI - 3D-QSAR studies of HDACs inhibitors using pharmacophore-based alignment. AB - Histone deacetylases (HDACs) enzyme is a promising target for the development of anticancer drugs. The enzyme-bound conformation of Trichostatin A (TSA) (PDB ID:1C3R) as an inhibitor of HDACs was used to manually construct a pharmacophore model. This model was then successfully used to identify the bioactive conformation and align flexible and structurally diverse molecules. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on hydroxamate-based HDACs inhibitors based on phamacophore alignment. The best predictions were obtained with CoMFA standard model (q(2) = 0.726, r(2) = 0.998) and CoMSIA model combined with steric, electrostatic, hydrophobic, hydrogen bond donor and acceptor fields (q(2) = 0.610, r(2) = 0.995). Both of the models were validated by an external test set, which gave a satisfactory predictive r(2) value of 0.800 and 0.732, respectively. Graphical interpretation of the results revealed important structural features of the inhibitors related to the active site of HDACs. The results may be exploited for further design and virtual screening for some novel HDACs inhibitors. PMID- 19136180 TI - Sedimentology models from activity concentration measurements: application to the "Bay of Cadiz" Natural Park (SW Spain). AB - A previous study on seabed sediments of the Bay of Cadiz (SW of Spain) enabled us to identify several relations between sedimentological variables and activity concentrations of environmental radionuclides such as (137)Cs, (226)Ra, (232)Th and (40)K. In this paper the study has been extended to a large neighbouring inter-tidal area in order to establish if the above mentioned models can be generalized. As a result we have determined that the measured activity concentrations are closely to the values predicted by the theoretical models (correlation coefficient range=0.85-0.93). Furthermore, the proposal model for granulometric facies as a function of activity concentrations of the abovementioned radionuclides provides for the sediments distribution a representation which agrees with the values of the tidal energy distribution obtained using numeric models calibrated with experimental data from current meters and water level recorders. PMID- 19136181 TI - Explaining the hip adduction moment variability during gait: Implications for hip abductor strengthening. AB - BACKGROUND: There is emerging interest in hip abductor function during gait and its potential relationship to knee joint pathology. During gait, the hip abductor muscles are primarily responsible for generating moments of force to control frontal plane movement. The current study investigated the relationship between hip abductor muscle function and frontal plane hip moments of force during gait. METHODS: Frontal plane hip moments of force and electromyographic features of gluteus medius were measured during walking in 22 healthy individuals. Hip abductor strength, subject anthropometrics and gait velocity were recorded. Multiple regression models were used to evaluate the relationship between the anthropometric, velocity, strength and electromyographic variables and the initial and mid-stance magnitude of the hip adduction moment. FINDINGS: A positive relationship was found between the initial peak moment (Nm), and both body mass and gait speed (R(2)=90%). Body mass (positive) and hip abductor strength (negative) explained significant levels of mid-stance magnitude variability (R(2)=62.5%). Gait speed (positive) explained significant levels of variability in the normalized initial peak moment (Nm/kg) (R(2)=52%). No variables were included in the normalized mid-stance moment model (P>0.05). INTERPRETATION: Body mass was the key factor associated with high hip adduction moments during initial and mid-stance of the gait cycle. Increased gait velocity was associated with higher initial peaks and higher muscle strength was associated with lower mid-stance magnitude of the external hip adductor moment during walking. These findings suggest that in a healthy adult population, hip abductor strength and activation were not directly related to the hip adduction moment magnitude during gait. PMID- 19136182 TI - Studying the infant feeding intentions of pregnant women experiencing material deprivation: methodology of the Looking at Infant Feeding Today (LIFT) study. AB - The Looking at Infant Feeding Today (LIFT) study examined the factors underlying the infant feeding choices of first-time mothers in the Leeds and Bradford areas of England experiencing material hardship, with a view to informing interventions aimed at increasing breastfeeding uptake amongst this group. This short report describes the novel methodology used to obtain data from a sample of pregnant women who are traditionally 'hard-to-reach' in deprived areas. This involved the use of trusted individuals such as midwives and multilingual health workers. A total of 449 women were approached; 441 (91.5%) agreed to participate and 303 returned completed questionnaires. Whilst 285 participants self-completed, 18 opted for interviewer assistance provided by a trained multilingual health worker. Feeding method up to 10 days after birth was obtained from the hospital records of 248 women (82.4% of eligible returns), and self-reported feeding method at six weeks was obtained from 273 women (90.7% of eligible returns). The study succeeded in obtaining data from a relatively deprived and hard-to-reach sample, with adequate numbers of participants in the subgroups of interest (teenage, left full time education age 16 or younger, household receiving income support or job seekers allowance, Asian or African-Caribbean). PMID- 19136183 TI - Community inequality and smoking cessation in New Zealand, 1981-2006. AB - The overall prevalence of smoking in New Zealand reduced from 32% in 1981 to 23.5% in 2006 but rates of smoking cessation have not been consistent among all social, demographic and ethnic groups. The period 1981-2006 also saw macroeconomic changes in New Zealand that resulted in profound increases in social and economic inequalities. Within this socio-political context we address two questions. First, has there been a social polarisation in smoking prevalence and cessation in New Zealand between 1981 and 2006? Second, to what extent can ethnic variation in rates of quitting be explained by community inequality, independently of socio-economic status? We find that smoking behaviour in New Zealand has become socially and ethnically more polarised over the past two decades, with greater levels of smoking cessation among higher socio-economic groups, and among New Zealanders of European origin. Variations in quit rates between Maori and European New Zealanders cannot be fully accounted for by ethnic differences in socio-economic status. Community inequality exerted a significant influence on Maori (but not European) smoking quit rates. The association with community inequality was particularly profound among women, and for particular age groups living in urban areas. These findings extend the international evidence for a relationship between social inequality and health, and in particular smoking behaviour. The research also confirms the importance of considering the role of contextual factors when attempting to elucidate the mechanisms linking socio-economic factors to health outcomes. Our findings emphasise that, if future smoking cessation strategies are to be successful, attention has to shift from policies that focus solely on engineering individual behavioural change, to an inclusion of the role of environmental stressors such as community inequality. PMID- 19136184 TI - Diversity within: deconstructing aboriginal community health in Wikwemikong Unceded Indian Reserve. AB - This paper analyzes the community health of the Wikwemikong Unceded Indian Reserve (WUIR), Ontario, Canada. Results are reported from fieldwork including participant observation, key informant interviewing and self-reported data measuring health status, risk behaviour, place of residence, self-identity, and personal history extracted from 350 interviews conducted during a community-wide needs assessment. The research aimed to create a health plan for the community; however, subsequent analysis of the needs assessment results indicates that internal diversity exists in health status and needs between the seven villages that comprise WUIR. The analysis suggests variation in health status and risk taking behaviour among community members may be related to varying colonial histories among the villages. The implications of intra-community variation in health status in First Nations are discussed in relation to influential health policy theories such as the determinants of health and health transfer policy in Canada. PMID- 19136185 TI - Practitioner perspectives on tackling health inequalities: findings from an evaluation of healthy living centres in Scotland. AB - Little is known about how health practitioners tasked with tackling health inequalities account for their own programmes and actions. This paper attempts to address this gap by drawing on data collected in the course of an evaluation of the Healthy Living Centre (HLC) programme, which was designed to address the wider determinants of health, in particular social exclusion and socioeconomic disadvantage, through targeting services at the most deprived local communities. Six Scottish HLC case studies explored in depth how HLC practitioners conceptualised 'health inequalities' and applied the construct to legitimate their public health and health improvement work. Practitioners drew on multiple explanations of health inequalities, sought to apply holistic approaches to service provision, and developed activities that took account of classed practices intended to overcome class-related disempowerment and stigma. They discussed the challenges of positioning services to appeal to and reach target groups and the difficulties in assessing the impact of their work on reducing health inequalities. Responses to tackling inequalities were variable across time and between HLCs, resulting from uneven learning about target groups and their changing needs, an evolving policy agenda and consideration given to the longer term sustainability of HLC sites. Although practitioners' work to address health inequalities was limited by the programme's focus on working with disadvantaged groups, findings illustrate how classed practices are linked to the challenges of attracting and successfully engaging with such groups. Practitioner accounts highlighted the importance of gaining acceptance to overcome barriers to engagement with disadvantaged communities, the time required to achieve a satisfactory level of engagement, the proximity of service providers to clients and the adaptability of services necessary to address evolving needs. PMID- 19136186 TI - Obesity, race/ethnicity and the multiple dimensions of socioeconomic status during the transition to adulthood: a factor analysis approach. AB - Racial/ethnic disparities in obesity widen dramatically during young adulthood in the US. Understanding racial/ethnic differences in the association between socioeconomic status (SES) and obesity can provide insight on these disparities. However, the delay and complexity of the transition to adulthood create challenges for defining SES using traditional, single indicators, such as income or years of education. Our objective was to define a multidimensional measure of young adult SES using exploratory factor analysis and to investigate whether distinct SES dimensions differentially predicted obesity across race/ethnicity in 11,250 young adults (mean age=21.9 years) from the National Longitudinal Study of Adolescent Health (Wave III: 2000-2001). Four factors (social advantage; schooling; employment; and economic hardship) extracted from a principal factor analysis on 38 SES indicators comprised our multidimensional measure of young adult SES. The respondents' scores on each factor were entered into gender stratified Poisson regression models to estimate the relative risk of young adult obesity for a contrast of approximately one standard deviation in score. The association of the "Social advantage" and "Economic hardship" factors with obesity differed by race/ethnicity (p<0.05 for Wald test of interaction) in females; high "Social advantage" scores were inversely associated with obesity in white and Hispanic females (9-20% lower) while high scores on "Economic hardship" were positively associated with obesity (7-76% higher) in white and Asian females. In contrast, no significant racial/ethnic differences were detected in young adult males. The "Schooling" factor was significantly protective (RR=0.91; 95% CI: 0.85, 0.98) for females of all racial/ethnic groups. These results facilitate understanding of the impact of multiple, distinct SES dimensions during the complex transition to adulthood and thus provide salient information for reducing racial/ethnic disparities in obesity during this important period for obesity development. PMID- 19136187 TI - Mechanisms behind privatization: a case study of private growth in Swedish elderly care. AB - For many years the Swedish Welfare State has been associated with a welfare model in which the public sector dominates both the provision and financing of the elderly care system. However, influenced by the ongoing trend of New Public Management, the past 15 years have been characterized by governmental regimes encouraging competition and as a result there has been a substantial increase in private providers. This case study on elderly care in Sweden provides new insights into the mechanisms behind the spread and growth of privatization. Our results show that in 1990 only 1% of the labour force in the elderly care sector was employed by private organizations, in comparison to 2003 when the private share had increased to 13%. The accompanying organizational changes have been controversial and are often criticized. In general, left-wing politicians have frequently defended the traditional welfare model dominated by public providers, whereas right-wing politicians have urged for a larger share of alternative providers. In this study, statistics between the years 1990 and 2003 were used to model the relationship between privatization and a number of economic, political and social/demographic variables. The results from regression and diffusion analysis imply that privately managed elderly care has established itself mainly in metropolitan areas dominated by right-wing regimes. Surprisingly, neighbouring municipalities tend to follow these pioneers irrespective of their political colour or economic situation. In fact, after shifting political power many of those neighbouring municipalities dominated by left-wing regimes not only maintain an abundance of private contractors but also encourage a continued process of contracting out publicly managed elderly care units. As a result, clusters of municipalities with an increasing degree of privatization arise despite political and economic differences. In conclusion, geographical proximity seems to be an important variable in addition to population density, ideology and financial situation when privatization reforms are implemented in the Swedish elderly care system. PMID- 19136188 TI - The role of religion in HIV-positive women's disclosure experiences and coping strategies in Kinshasa, Democratic Republic of Congo. AB - Literature from the U.S. has documented the importance of spirituality on the psychological health of people living with HIV/AIDS; however there is little published data on the ways in which people living with HIV/AIDS in Africa turn to religion for support. We conducted 40 in-depth interviews with HIV-positive women who were pregnant or had recently given birth in Kinshasa, Democratic Republic of Congo to inform the development of a comprehensive family-centered HIV treatment and care program. Women described how they relied upon their faith and turned to church leaders when they were diagnosed with HIV and prepared to share their diagnosis with others. The women used prayer to overcome the initial shock, sadness and anger of learning their HIV diagnosis. They turned to their church leaders to help them prepare for disclosing their diagnosis to others, including their partners. Church leaders were also important targets for disclosure by some women. Women's faith played an important role in their long-term coping strategies. Conceptualizing their infection as a path chosen by God, and believing that God has the power to cure their infection comforted women and provided them with hope. In settings like the Democratic Republic of the Congo, where there is a strong foundation of faith, we need to recognize how individuals draw upon their different health belief systems in order to develop and implement coherent and effective prevention, treatment and care strategies. PMID- 19136189 TI - Is perceived racial privilege associated with health? Findings from the Behavioral Risk Factor Surveillance System. AB - While racial discrimination has gained increasing attention in public health research, little is known about perceived racial privilege and health. Using the Behavioral Risk Factor Surveillance System (BRFSS) data, this study explored the relationship of both perceived racial discrimination and privilege with well being in the USA. Data were extracted from the BRFSS 2004 data set, in which 22,412 respondents in seven states and one major city provided data on perceived racial discrimination and privilege at work. Logistic regression analysis was conducted to examine the relationships of differential racial treatment to self rated general health status and the number of physically and mentally unhealthy days. Racially stratified analyses found that perceived racial privilege was significantly associated with more days of poor physical and mental health. This relationship was consistent for Whites, but for racial minorities it appeared on only some outcome measures. Reports of being treated worse than other races in the workplace were associated with poor health for all racial groups, as had been reported in previous studies on racial discrimination. Because racial discrimination and racial privilege are both products of racism, this study's findings suggest that racism may harm all involved. Impacts of perceived racial privilege deserve more attention in the literature on racism and health. PMID- 19136190 TI - "When in the body, it makes you look fat and HIV negative": the constitution of antiretroviral therapy in local discourse among youth in Kahe, Tanzania. AB - Antiretroviral therapy (ART) is becoming increasingly more accessible within the health care system in Tanzania. However, the impact of the increased availability of ART on local conceptions about medicines, health and physical wellbeing has not been fully explored. In this article we examine how ART is constituted within local discourses about treatment and healing. Based on 21 focus group discussions with young people aged 14-24 years in a rural area (Kahe), we examine how local terms and descriptions of antiretroviral therapy relate to wider definitions about the body, health, illness and drug efficacy. Findings illustrate how local understandings of ART draw on a wider discourse about the therapeutic functions of medicines and clinical dimensions of HIV/AIDS. Therapeutic efficacy of antiretroviral medication appeared to overlap and sometimes contradict locally shared understandings of the clinical functions of medicines in the body. Implications of ART on bodily appearance and HIV signs may influence conceptions about sick role, perpetuate stigma and affect local strategies for HIV prevention. Structural inequities in access, limited information on therapeutic efficacy of ART and perceived difficulties with status disclosure appear to inform local conceptions and possible implications of ART. Policy and programme interventions to foster public understanding and acceptability of ART should emphasize treatment education about the benefits and limitations of therapy and increased access to ART in rural areas, and should integrate voluntary status disclosure and HIV prevention. PMID- 19136191 TI - A combination of umbilical artery PI and normalized blood flow volume in the umbilical vein: venous-arterial index for the prediction of fetal outcome. AB - OBJECTIVE: The objective was to assess the diagnostic power of the umbilical venous-arterial index (VAI) as a combination of the pulsatility index in the umbilical artery and the normalized blood flow volume in the umbilical vein for the prediction of poor fetal outcome. STUDY DESIGN: This was a prospective clinical study in which the umbilical artery PI (UAPI), the normalized umbilical vein blood volume flow rate (nUV; ml/min/kg estimated fetal body weight), the venous-arterial index (VAI; nUV/UAPI), and the pulsatility index (PI) in the umbilical artery (UA), uterine artery (utA), middle cerebral artery (MCA), and aorta were determined in 181 fetuses once (at between 17 and 41 weeks' gestation) during pregnancy using standard Doppler ultrasound equipment. A risk score based on the umbilical blood pH, the 1 min Apgar score, birth weight, duration of gestation, type of postpartum respiratory support, and referral to the pediatric department was used, and fetuses were assigned to a control or a pathological group accordingly. RESULTS: The incidence of compromised neonates was 18.2%. The sensitivity of the UAPI in predicting the poor neonatal outcome was 51.5%, of the nUV 54.5%, the MCA PI 39.4%, the PI in the utA 61.5%, and the notching in the utAa and the VAI was 57.6% and 69.7% respectively. CONCLUSION: A combination of the umbilical artery PI and the nUV as the VAI with a cut-off of 100 ml/min/kg can be used to predict fetal outcome. PMID- 19136193 TI - Port-site metastasis following laparoscopic hysterectomy and bilateral salpingo ophorectomy for endometrial carcinoma. PMID- 19136192 TI - Severe maternal morbidity for 2004-2005 in the three Dublin maternity hospitals. AB - OBJECTIVE: To assess the prevalence and causes of severe maternal morbidity in Dublin over a two year period from 2004 to 2005. STUDY DESIGN: A prospective cohort study from January 2004 to December 2005 was undertaken in the three large maternity hospitals in Dublin, which serve a population of 1.5 million people. All are tertiary referral centres for obstetrics and neonatology and have an annual combined delivery rate of circa 23,000 births. Cases of severe maternal morbidity were identified. A systems based classification was used. The primary cause of maternal morbidity and the number of events experienced per patient was recorded. RESULTS: We identified 158 women who fulfilled the definition for severe maternal morbidity, giving a rate of 3.2 per 1000 maternities. There were two maternal deaths during the time period giving mortality to morbidity ratio of 1:79. The commonest cause of severe morbidity was vascular dysfunction related to obstetric haemorrhage. Eclampsia comprised 15.4% of cases. Intensive care or coronary care admission occurred in 12% of cases. CONCLUSION: The prevalence of severe maternal morbidity in this population is 3.2/1000 maternities. Obstetric haemorrhage was the main cause of severe maternal morbidity. PMID- 19136194 TI - Sociodemographic factors and pregnancy events associated with women who declined vaginal birth after cesarean section. AB - OBJECTIVE: Certain sociodemographic characteristics and pregnancy events were shown to pre-dispose women to decline vaginal birth after cesarean section (CS) in the Western population. The study aimed to identify the sociodemographic factors and pregnancy events associated with Chinese women who declined to attempt vaginal birth after CS. STUDY DESIGN: This was a retrospective study of Hong Kong Chinese women who delivered in a public obstetric unit between 2002 and 2006. It was the unit's policy that women should attempt vaginal birth after one previous lower segment CS. During the study period, 1317 healthy Hong Kong Chinese women carrying a singleton pregnancy had had one previous lower segment CS. Of these women, 787 had had no antenatal complications during their index pregnancy. Univariate and multivariate analyses were performed to identify the sociodemographic factors and pregnancy events associated with women who declined vaginal birth after CS. RESULTS: Thirty-nine percent (39.26%; 95%CI 35.85, 42.67) of women had elective CS because they had declined a vaginal birth. A history of a previous successful vaginal birth was the strongest determinant negatively associated with declining a vaginal birth (OR 0.15; 95%CI 0.06, 0.37). Determinants that were positively associated with declining a vaginal birth were higher family social class (OR 1.61; 95%CI 1.02, 2.55); family income of more than US$ 3850 per month (OR 1.56; 95%CI 1.03, 2.34), and women who attended private antenatal care before their first visit to the public unit (OR 1.47; 95%CI 1.02, 2.12). CONCLUSIONS: Women who experienced a previous successful vaginal birth were positively associated with attempting vaginal birth after CS. Women who declined vaginal birth after CS exhibited certain social characteristics and treatment-seeking behavior. Future studies into the social context of this group of women may help us understand the underlying reasons why they decline vaginal birth. PMID- 19136195 TI - Studies on equilibrium of anthranilic acid in aqueous solutions and in two-phase systems: aromatic solvent-water. AB - The acid-base equilibria of anthranilic acid have been characterized in terms of macro- and microdissociation constants (dissociation constants K(a1), K(a2) and tautomeric constant K(z)). On the basis of spectrophotometric investigations the values of the distribution ratio D of anthranilic acid in the two-phase systems: aromatic solvent (benzene, ethylbenzene, toluene, chlorobenzene, bromobenzene) aqueous solution were obtained. Employing the results of potentiometric titration in the two-phase systems: aromatic solvent-aqueous solution the distribution constant K(D) and dimerization constant K(dim) values were calculated. The influence of organic solvent polarity and pH of the aqueous phase on the contents of the particular forms of the acid in the two-phase systems were analyzed. PMID- 19136196 TI - Defective cytotoxicity of T lymphocytes in myelodysplastic syndrome. AB - OBJECTIVE: The capacity of mononuclear blood cells to form autoreactive cytotoxic T lymphocytes was investigated in order to elucidate the mechanism of successful immunosuppressive therapy in some myelodysplastic syndrome (MDS) patients (autoreactivity studies). The failure in autoreactivity studies raised the question of alloreactive cytotoxic T lymphocyte formation in MDS (alloreactivity studies). MATERIALS AND METHODS: Sixteen MDS patients and relevant controls were examined. Autoreactive lymphocytes directed against autologous bone marrow mononuclear cells and alloreactive lymphocytes directed against unrelated third party cells were tested using cytotoxicity assay. In addition, we used one-way mixed lymphocyte reaction, human androgen receptor test for clonality detection, and enzyme-linked immunosorbent assay kits for tumor necrosis factor and interferon-gamma testing. RESULTS: We did not confirm the presence of autoreactive T cells in eight of nine MDS patients tested. The response to allogeneic cells was impaired in 11 of 16 MDS patients, more often in refractory anemia (RA; 80%) than in RA with ring sideroblasts (40%). Interestingly, the response to allogeneic cells in mixed lymphocyte reaction was normal in all MDS patients. T lymphocytes were polyclonal in all but one patient. Tumor necrosis factor and interferon-gamma level in supernatants of mononuclear cells was significantly reduced in RA. CONCLUSION: The presumed autoaggressive T cells were not confirmed in MDS in our experimental arrangement. Alloreactivity studies demonstrated the impairment of effector cytotoxic phase of cell-mediated immunological reaction in MDS, namely in RA. The significance of our finding of defective cytotoxicity for pathogenesis, clinical course, and even for therapy is discussed together with other immunological defects reported so far. PMID- 19136197 TI - On the contribution of modelling to multifunctional agriculture: learning from comparisons. AB - In this paper a set of criteria is proposed for the evaluation of the potential contribution of modelling tools to strengthening the multifunctionality of agriculture. The four main areas of evaluation are (1) policy relevance, (2) the temporal resolution and scope, (3) the degree to which spatial and socio institutional scales and heterogeneity are addressed and (4) the level of integration in the assessment of scientific dimensions and of the multiple functions of agriculture. The evaluative criteria are applied to the portfolio of modelling approaches developed and applied in a joint project of the French research institute INRA and the Dutch Wageningen University & Research Centre. The CLUE-S model focuses on prediction of changes in multifunctional land-use at regional scale, given a set of predetermined scenarios or policy variants, e.g. for ex-ante policy assessment and initiation of discussions on regional development. The two other modelling approaches are complementary and aim to address multifunctional farming activities. The Landscape IMAGES framework generates a range of static images of possible but sometimes distant futures for multifunctional farming activities in a small region or landscape. It supports the exploration of trade-offs between financial returns from agriculture, landscape quality, nature conservation and restoration, and environmental quality. Co-Viability Analysis generates trajectories of states and farming decisions fulfilling a given set of ecological and productive constraints representing a desired and sustainable future. The three modelling approaches differ in their policy relevance, in the ways that spatial and socio institutional scales are addressed and in their degree of explicitation of interaction between the various functions of agriculture, but jointly cover most of the desired capabilities for assessment of multifunctionality. Caveats were particularly identified in the integration of the socio-institutional dimension and the related heterogeneity. Although the model portfolio did not completely satisfy the demands of the set of evaluative criteria, it is concluded that, due to their complementarities, in combination the three models could significantly contribute to further development and strengthening of multifunctionality. PMID- 19136198 TI - Editorial comment on: prognosis of Japanese metastatic renal cell carcinoma patients in the cytokine era: a cooperative group report of 1463 patients. PMID- 19136199 TI - Prognosis of Japanese metastatic renal cell carcinoma patients in the cytokine era: a cooperative group report of 1463 patients. AB - BACKGROUND: Incidence rate of renal cell carcinoma (RCC) differs among countries. The rates of Asian countries are lower than those of countries in North America or Europe but are exceptionally high in Japanese males. Approximately 30% of patients with RCC have metastasis at initial diagnosis, and another 30% have metastasis after nephrectomy. Clinical studies of risk factors in patients with metastatic RCC (mRCC) are mainly based on data from non-Asian patients. OBJECTIVES: We aimed to investigate the prognosis of Japanese patients and their prognostic factors. DESIGN, SETTING, AND PARTICIPANTS: The subjects of this study were 1463 patients who were clinically diagnosed with RCC with metastasis in 40 Japanese hospitals between January 1988 and November 2002. MEASUREMENTS: The primary end point was overall survival calculated from first diagnosis of mRCC to death or last follow-up. We also investigated the relationship between survival and clinical features. RESULTS AND LIMITATIONS: The median overall survival time was 21.4 mo. The estimated survival rates at 1, 3, 5, and 10 yr were 64.2%, 35.2%, 22.5%, and 9.1%, respectively; they contrasted with data from the United States of 54%, 19%, 10%, and 6%, respectively for the same periods. A high percentage of patients had undergone nephrectomy (80.5%) and metastasectomy (20.8%), both of which were shown to prolong survival. CONCLUSIONS: The median survival time in the present study was approximately twice as long as that of previous studies from North America or Europe. Early diagnosis of metastasis, nephrectomy, metastasectomy, and cytokine-based therapy seemed to improve the prognosis of RCC patients in the present study. PMID- 19136200 TI - Process in the mechanisms of endostatin combined with radiotherapy. AB - Endostatin has been demonstrated to represent a promising novel medicine to treat malignant tumors, and it may be more effective when combined with conventional treatment. Meanwhile, a promising area of research in radiation oncology is the integration of molecular targeting agents to improve the effectiveness of radiotherapy (RT) in the control of primary tumor. Antiangiogenic agents are one such class of targeted therapies and have shown promise in both laboratory and clinical experiments. Endostatin in combination with RT has feasibility; but it also has lots of problems. Further understanding of the mechanisms is needed. PMID- 19136201 TI - Highly homologous hS100A15 and hS100A7 proteins are distinctly expressed in normal breast tissue and breast cancer. AB - Human S100A7 (psoriasin) is considered a marker for specific stages of breast cancer. hS100A15 is almost identical to hS100A7 and difficult to discriminate. We developed specific probes to distinguish hS100A7 and hS100A15, and demonstrate their differential distribution in normal breast tissue. Further, hS100A7 and S100A15 transcripts are elevated in ER/PR negative breast cancers, but hS100A15 protein is detected in all cancer specimens while hS100A7 protein is sporadically expressed. The differential regulation, expression and distribution of hS100A7 and hS100A15 and their reported distinct functions are compelling reasons to discriminate among these proteins in normal breast and breast cancers. PMID- 19136202 TI - Adsorption removal of phosphate in industrial wastewater by using metal-loaded skin split waste. AB - Leather industry inevitably generates a large amount of skin split waste (SSW) due to the necessary operation of splitting in leather making process. In this study, two adsorbents, SSW-Fe and SSW-Al, were prepared by loading Fe(III) and Al(III) onto SSW, and their adsorption behaviors to phosphate in industrial wastewater were investigated through batch and column adsorption experiments. The macro-adsorption kinetics data obtained from batch experiments were well fitted by the pseudo-second-order rate model and the adsorption isotherms can be well described by the Langmuir equation. The adsorption behaviors of the columns can be well described by Yoon and Nelson model and the time at breakthrough point can be accurately predicted by this model. It was found that the metal-loaded adsorbents prepared by using SSW as supporting matrix can effectively remove phosphate from industrial wastewater. The leakage of metal ions during adsorption process is neglectable. These results indicate that the metal-loaded SSW adsorbents have a good future in practical application for the removal of phosphate from industrial wastewater. PMID- 19136203 TI - Sources and reactivity of NMHCs and VOCs in the atmosphere: a review. AB - Nonmethane hydrocarbons (NMHCs) and volatile organic compounds (VOCs) are important species present in the environment, which results in alteration of the chemistry of atmosphere. On the global scale natural emissions of NMHCs and VOCs exceed anthropogenic emissions, although anthropogenic sources usually dominate within urban areas. Among the natural sources, vegetation is the dominant source. Oceanic and microbial production of these species is minimal as compared to other sources of input. Isoprene and terpenes are main species of NMHCs which are emitted from plants as a protective mechanism against temperature stress tolerance and protection from ravages of insects and pests. The major anthropogenic sources for NMHCs emissions are biomass burning and transportation. NMHCs play a significant role in ozone (O(3)) production in the presence of adequate concentration of oxides of nitrogen in the atmosphere. The production of O(3) is based on Maximum Incremental Reactivity (MIR) of NMHCS and VOCs. The compound's MIR multiplied by molecular weight gives Relative Ozone Productivity (ROPi). To check the reliability of current methods of measuring the NMHCs the Nonmethane Hydrocarbon Inter-comparison Experiment (NMHICE) had been designed. The sample of known composition and unknown concentration of different hydrocarbons was supplied to different laboratories worldwide and less than 50% laboratories correctly separated the unknown mixture. Atmospheric scientists throughout the world are evaluating current analytical methods being employed and are trying to correct the problems to ensure quality control in hydrocarbon analysis. PMID- 19136204 TI - Chromatographic separations and recovery of lead ions from a synthetic binary mixtures of some heavy metal using cation exchange resin. AB - A simple solid phase extraction procedure on cation exchange resin Purolite C100 is presented. The procedure based on a column technique for separation and recovery of lead ions from synthetic binary mixtures. Equilibrium distribution coefficient, k(d) for the different metal ions such as Al(III), Fe(III), Ba(II) and Pb(II) in the presence of nitric acid and ammonium acetate solutions of variable concentrations was determined at 25 degrees C. The values of separation factor, alpha were evaluated. Quantitative separation of lead ions from a synthetic binary mixtures are based on the fact that ammonium acetate is a good elute for lead ions but fails to elute the other cations. PMID- 19136205 TI - Kinetics and mechanism of photolysis and TiO2 photocatalysis of triclosan. AB - The degradations of triclosan (5-chloro-2-(2,4-dichlorophenoxy)-phenol), a potent broad-spectrum antimicrobial agent, were compared in TiO2-only in the dark condition, photolysis, and TiO2 photocatalysis with a UV-A lamp. TiO2 photocatalysis more effectively degraded and mineralized triclosan compared to TiO2-only and photolysis conditions. While triclosan removed only 30% by TiO2 only condition within 20 min, the triclosan degradation in photolysis and photocatalysis at the same time was 75 and 82%, respectively, and TOC removal was significantly higher in photocatalysis than in photolysis. The data of kinetics showed that triclosan adsorption onto TiO2 was fitted to Langmuir isotherm, and TiO2 photocatalysis was fitted to Langmuir-Hinshelwood model (b=27.99 mM(-1), K(triclosan)=9.49 mM(-1)). The neutral range of pH was favorable to photocatalysis due to the charge effect between TiO2 and triclosan. The addition of 2-propanol, a radical scavenger, significantly reduced the degradation of triclosan both in photolysis and photocatalysis. Dioxin-type intermediates such as dibenzo-dichloro-p-dioxin (DCDD), dibenzo-p-dioxin were produced in photolysis with and without 2-propanol, and also in photocatalysis with 2-propanol, but these intermediates were not detected in photocatalysis without 2-propanol. This result indicates that the photocatalytic degradation of triclosan is mainly achieved by radicals, and these radicals can further degrade dioxin-type intermediates once they are produced in photocatalysis. PMID- 19136206 TI - Theoretical studies on polynitro-1,3-bishomopentaprismanes as potential high energy density compounds. AB - Based on the fully optimized molecular geometric structures at the DFT-B3LYP/6 31G* level, the densities (rho), detonation velocities (D) and pressures (P) for a series of polynitro-1,3-bishomopentaprismanes (PNBPPs), as well as their thermal stabilities, were investigated to look for high energy density compounds (HEDCs). The studied PNBPPs have high values of heats of formation (HOFs) and the magnitude is correlative with the number (n) and the space distance of nitro groups. D and P for PNBPPs were estimated by using modified Kamlet-Jacobs equations based on the calculated HOFs and rho. It is found that rho, D and P all increase with n and satisfy the group additivity rule. The calculations on the bond dissociation energies of C_NO(2) and C_C bonds indicate that both bonds are possible to be the trigger bond in the pyrolysis process, and this interesting phenomenon is related with the molecular structure, especially the strain energy of the skeleton. In conjunction with the energetic performances and thermal stabilities, PNBPPs with n=8-12 are recommended as the preferred candidates of HEDCs. These results would provide basic information for the further studies of PNBPPs. PMID- 19136207 TI - Feasibility analysis of color removal from textile dyeing wastewater in a fixed bed column system by surfactant-modified zeolite (SMZ). AB - In this study, the ability of surfactant-modified zeolite (SMZ) to remove color from real textile wastewater was investigated. Tests were performed in a fixed bed column reactor and the surface of natural zeolite was modified with a quaternary amine surfactant hexadecyltrimethylammonium bromide (HTAB). The zeolite bed that was modified at 1 g L(-1) HTAB concentration and HTAB flow rate of 0.015 L min(-1) showed good performance in removing color. Effects of wastewater color intensity, flow rates and bed heights were also studied. Wastewater was diluted several times in the ratios of 25%, 50% and 75% in order to assess the influence of wastewater strength. The breakthrough curves of the original and diluted wastewaters are dispersed due to the fact that breakthrough came late at lower color intensities and saturation of the bed appeared faster at higher color intensities. The column had a 3-cm diameter and four different bed heights of 12.5, 25, 37.5 and 50 cm, which treated 5.25, 19.50, 35.25 and 51 L original textile wastewater, respectively, at the breakthrough time at a flow rate of 0.025 L min(-1). The theoretical service times evaluated from bed depth service time (BDST) approach for different column variables. The calculated and theoretical values of the exchange zone height were found with a difference of 27%. The various design parameters obtained from fixed-bed experimental studies showed good correlation with corresponding theoretical values, under different bed heights. The regeneration of the SMZ was also evaluated using a solution consisting of 30 g L(-1) NaCl and 1.5 g L(-1) NaOH at pH 12 and temperature 30 degrees C. Twice-regenerated SMZ showed the best performance compared with the others while first- and thrice-regenerated perform lower than the original SMZ. PMID- 19136208 TI - Removal of phosphate from secondary effluent with Fe2+ enhanced by H2O2 at nature pH/neutral pH. AB - Removal of phosphate in secondary effluents was investigated in presence of Fe(2+)/H(2)O(2). The effect of H(2)O(2)-dose, Fe-dose and initial phosphate concentration were assessed. The results indicated that Fe(2+)/H(2)O(2) could greatly increase the removal of phosphate compared with those achieved by Fe(2+) alone. For initial phosphate concentration of 0.97-3.75 mg P/L, phosphate removal rates of 50-60% were observed at 1:1 molar addition of Fe(II). However, a 125% excess of Fe-dose was necessary for complete phosphate removal. The experimental data demonstrated that removal of phosphate with Fe(2+)/H(2)O(2) was higher than that observed with ferric coagulation alone. This fact suggested that in situ formed Fe(III) having much affinity for ligand phosphate. Chemical co precipitation was considered as the dominant mechanism about phosphate removal in presence of Fe(2+)/H(2)O(2). The electron paramagnetic resonance (EPR) spectra tests in secondary effluents showed that Fe(2+)/H(2)O(2) could produce an increasing hydroxyl radical concentration with a decrease in both H(2)O(2) dosage and phosphate concentration. Fe(2+)/H(2)O(2) had the potential to be utilized for removal of phosphate due to the lower cost and the higher phosphate removal capability. PMID- 19136209 TI - Thermal behaviour of arsenic trioxide adsorbed on activated carbon. AB - The thermal stability and desorption of arsenic trioxide (As(2)O(3)) adsorbed on activated carbon (AC) was investigated as this phenomenon is expected to influence the arsenic release during low temperature pyrolysis of chromated copper arsenate (CCA) wood waste. Firstly, a thermogravimetric (TG) experiment with arsenolite, an allotropic form of As(2)O(3), was performed. The sample starts to sublime at temperatures lower than 200 degrees C with a sublimation peak temperature of 271 degrees C. Subsequently, TG experiments with samples of As(2)O(3) adsorbed on AC revealed that only very little (max. 6+/-3 wt%) As(2)O(3) was volatilized at temperatures below 280 degrees C, while still 41.6 (+/-5)wt% of the original arsenic concentration was retained at 440 degrees C and 28.5 (+/-3)wt% at 600 degrees C. The major arsenic volatilization occurred between 300 degrees C and 500 degrees C. The kinetic parameters of desorption, activation energy of desorption (E(d)) and pre-exponential factor (A), were determined by fitting an Arrhenius model to the experimental data, resulting in E(d)=69 kJ/mol, A=1.21 x 10(4)s(-1). It can be concluded that the adsorption of As(2)O(3) on AC can contribute to the thermal stabilisation of As(2)O(3). Consequently, during low temperature pyrolysis of CCA wood arsenic release may be prevented by adsorption of As(2)O(3) on the coal-type product formed during the thermal decomposition of the wood. PMID- 19136210 TI - Modeling of gadolinium recovery from nitrate medium with 8-hydroxyquinoline by emulsion liquid membrane. AB - The extraction equilibrium of Gd(III) from nitrate medium by 8-hydroxyquinoline (HOX) in toluene was studied. Liquid-liquid investigations were first carried out. Based on the equilibrium results, the extraction of Gd(III) from aqueous nitrate medium into an emulsion liquid membrane system (ELM) containing 8 hydroxyquinoline in toluene as extractant, HNO(3) as stripping solution, Span-80 as surfactant was studied. The stability of the prepared ELM was studied in terms of the degree of membrane breakage. The different parameters affecting the permeation of gadolinium (III) were also studied. A general permeation model for the recovery of Gd(III) by the selected membrane is presented. The internal mass transfer in the water in oil (W/O) emulsion drop, the external mass transfer around the drop, the rates of formation and decomposition of the complex at the external aqueous-organic interface were considered. PMID- 19136211 TI - Area-specific representation of mechanical nociceptive stimuli within SI cortex of squirrel monkeys. AB - While functional imaging studies in humans have consistently reported activation of primary somatosensory cortex (SI) with painful stimuli, the specific roles of subdivisions of areas 3a, 3b, and 1 within SI during pain perception are largely unknown, particularly in the representation of mechanical evoked pain. In this study, we investigated how modality, location, and intensity of nociceptive stimuli are represented within SI by using high-spatial resolution optical imaging of intrinsic signals in Pentothal-anesthetized squirrel monkeys. Perceptually comparable mechanical nociceptive and innocuous tactile stimuli were delivered by indenting the glabrous skin of the distal finger pads with 0.2 and 2mm diameter probes, respectively. Within each of areas 3a, 3b, and 1, activations to mechanical nociceptive stimulation of individual distal finger pads were spatially distinct and somatotopically organized. We observed differential cortical activation patterns. Areas 3a, 3b, and 1 were all activated during mechanical nociceptive stimulation and were modulated by nociceptive stimulus intensity. However, with innocuous tactile stimulation, mainly areas 3b and 1 exhibited response modulation with different levels of stimulation. In summary, mechanical nociceptive inputs are area-specific and topographically represented within SI. We propose that all areas of SI are implicated in encoding the features of mechanical nociception, where areas 3a and 3b are distinctively involved in coding nociceptive and pressure sensation components of stimulation. PMID- 19136212 TI - Spin polarization of field-emitted electrons from half-metallic Co2MnSi thin films grown on a W(001) facet. AB - We measured spin polarization of electrons field-emitted from half-metallic Co(2)MnSi thin film grown on a W(001) facet via chromium buffer layer using Mott scattering. For spontaneously magnetized samples, values of polarization at room temperature were observed in a range from 10% to 46% and the polarization direction was 110 orientation of substrate tungsten, which agreed with an easy axis of magnetization of bulky Co(2)MnSi. An enhancement of polarization was observed as a consequence of applying a magnetic field of 350G perpendicular to the emitter axis after the annealing at 800K. This result is considered to be caused by improvement in crystallinity of the evaporated film due to annealing. PMID- 19136213 TI - Atom probe analysis of titanium hydride precipitates. AB - It is expected that the three-dimensional atom probe (3DAP) will be used as a tool to visualize the atomic scale of hydrogen atoms in steel is expected, due to its high spatial resolution and very low detection limit. In this paper, the first 3DAP analysis of titanium hydride precipitates in metal titanium is reported in terms of the quantitative detection of hydrogen. FIB fabrication techniques using the lift-out method have enabled the production of needle tips of hydride precipitates, of several tens of microns in size, within a titanium matrix. The hydrogen concentration estimated from 3DAP analysis was slightly smaller than that of the hydride phase predicted from the phase diagram. We discuss the origin of the difference between the experimental and predicted values and the performance of 3DAP for the quantitative detection of hydrogen. PMID- 19136214 TI - Molecular characterization of Blastocystis isolates from children and rhesus monkeys in Kathmandu, Nepal. AB - To investigate the possible transmission of Blastocystis organisms between local rhesus monkeys and children in Kathmandu, Nepal, we compared the subtype (ST) and sequence of Blastocystis isolates from children with gastrointestinal symptoms and local rhesus monkeys. Twenty and 10 Blastocystis isolates were established from 82 and 10 fecal samples obtained from children and monkeys, respectively. Subtype analysis with seven sequence-tagged site (STS) primers indicated that the prevalence of Blastocystis sp. ST1, ST2 and ST3 was 20%, 20% and 60% in the child isolates, respectively. In contrast to human isolates, ST3 was not found in monkey isolates and the prevalence of ST1 and ST2 was 50% and 70%, respectively, including three mixed STs1 and 2 and one isolate not amplified by any STS primers, respectively. Since Blastocystis sp. ST2 has been reported as the most dominant genotype in the survey of Blastocystis infection among the various monkey species, sequence comparison of the 150bp variable region of the small subunit rRNA (SSU rRNA) gene was conducted among ST2 isolates of humans and monkeys. Sequence alignment of 24 clones developed from ST2 isolates of 4 humans and 4 monkeys showed three distinct subgroups, defined as ST2A, ST2B and ST2C. These three subgroups were shared between the child and monkey isolates. These results suggest that the local rhesus monkeys are a possible source of Blastocystis sp. ST2 infection of humans in Kathmandu. PMID- 19136215 TI - Beyond RECIST: molecular and functional imaging techniques for evaluation of response to targeted therapy. AB - The development of targeted therapies is a major breakthrough in the treatment of cancer. By evoking necrosis and cavitation, evaluation based on tumour size alone, as is done in the RECIST criteria, is no longer an adequate method. New molecular and functional imaging techniques are developed. This review focuses on the use of new imaging modalities for the evaluation of treatment response of pathway based targeted therapies. First, the basic principles of functional and molecular imaging modalities are briefly discussed. Thereafter, their clinical application in targeted therapies is correlated to the underlying biological mechanism. In this way, the best method for response evaluation for a new agent can be identified. PMID- 19136216 TI - Amygdala activity to fear and anger in healthy young males is associated with testosterone. AB - Neuroimaging studies have documented modulation of the activity of the amygdala - a key node in the neural network underlying emotion perception and processing, and one that has also been associated with regulating aggression - by exogenous testosterone. However, results on the impact of normal range testosterone levels on explicit emotion recognition as a prerequisite for social interaction and amygdala activation in healthy young males are missing. Hence, we performed functional MRI at 3T in a group of 21 healthy males during explicit emotion recognition with a protocol specifically optimized to reliably detect amygdala activation. We observed similar amygdala activation to all emotions presented without any effect of gender of poser or laterality. Reaction times to fearful male faces were found negatively correlated to testosterone concentration, while no significant effects emerged for other emotions and neutral expressions. Correlation analyses revealed a significant positive association between testosterone levels and amygdala response to fearful and angry facial expressions, but not to other expressions. Hence, our results demonstrate that testosterone levels affect amygdala activation and also behavioral responses particularly to threat-related emotions in healthy young males. We conclude that these findings add to our understanding of emotion processing and its modulation by neuroendocrine factors. PMID- 19136217 TI - Schizotypy and nicotine, alcohol, and cannabis use in a non-psychiatric sample. AB - Schizotypy is a multidimensional personality construct that is characterized by perceptual abnormalities, social withdrawal, mild suspiciousness, and odd thinking patterns. This study examined the relationship between four dimensions of self-reported schizotypy and substance use involving nicotine, alcohol, and cannabis, in undergraduate students. Results showed that higher levels of disorganized schizotypy, or odd thinking and behavior, were associated with greater indices of use of all three substances. Furthermore, higher cognitive perceptual schizotypy was selectively associated with cannabis use. Results confirm findings of recent research that has discovered associations among schizotypy and substance use, highlighting links between behavioral traits and use of nicotine, alcohol, and cannabis. This study is one of the first to examine a wide range of schizotypy domains, and to show selective effects of the disorganized domain of schizotypy. PMID- 19136218 TI - Pre-colonized polymeric vascular stents: a hope for atherosclerotic vascular disease. PMID- 19136219 TI - Developmental neural chromosome instability as a possible cause of childhood brain cancers. PMID- 19136220 TI - Is cancer a genetic program with an unknown function? AB - Cancer is a tissue that requires a genetic program, possibly involving hundreds of genes, as indicated by studies of differential gene expression performed with microarrays in various types of cancer. Cancer is thus a biological process of considerable precision as described by Schrodinger's equation, and it is not simply the random expression of hundreds of genes, but rather a program that gives neoplastic tissue characteristics that are independent of the originating tissue: invasion capacity and destruction of neighbouring tissues, metastasis, and capacity to progressively disengage from the genetic program of the original tissue. This program is common to all types of cancer, and does not appear to be related to the genetic program of any known differentiated tissue. In other words, cancer's genetic program appears to be universal in all tissues, since cancer behaves similarly in all tissues; though depending on the tissue in which it originates it will show certain specific characteristics, reflecting the extent to which the neoplastic cells are able to escape from the characteristics of the original tissue. PMID- 19136222 TI - Modeling risk factors for nicotine and other drug abuse in the preclinical laboratory. AB - Risk factors that predict vulnerability for nicotine and other drug abuse have been identified using preclinical models, and there is close agreement with clinical and epidemiological studies. The major risk factors to be discussed are age, sex/hormonal status, impulsivity, sweet-liking, novelty reactivity, proclivity for exercise, and environmental impoverishment (vs. enrichment). This discussion will focus on factors that preclinical research has determined are strong and translatable predictors of nicotine and other drug abuse. An advantage of using preclinical models is that prospective, longitudinal studies and within subject designs can be used to reveal risk factors that are diverse yet maintain unique characteristics. The many interrelationships among these factors lead to an additive vulnerability that increases the predictability that drug abuse will occur. A feature that these risk factors have in common is that they consistently predict vulnerability to drug abuse over critical transition phases of addiction that are difficult to examine prospectively in humans, such as acquisition, escalation, and reinstatement of drug-seeking after abstinence (relapse). The models offer valuable information that has been transferred to effective prevention and treatment strategies for smoking and other drug abuse in humans. PMID- 19136221 TI - Tissue pO2 of orthotopic 9L and C6 gliomas and tumor-specific response to radiotherapy and hyperoxygenation. AB - PURPOSE: Tumor hypoxia is a well-known therapeutic problem; however, a lack of methods for repeated measurements of glioma partial pressure of oxygen (pO(2)) limits the ability to optimize the therapeutic approaches. We report the effects of 9.3 Gy of radiation and carbogen inhalation on orthotopic 9L and C6 gliomas and on the contralateral brain pO(2) in rats using a new and potentially widely useful method, multisite in vivo electron paramagnetic resonance oximetry. METHODS AND MATERIALS: Intracerebral 9L and C6 tumors were established in the left hemisphere of syngeneic rats, and electron paramagnetic resonance oximetry was successfully used for repeated tissue pO(2) measurements after 9.3 Gy of radiation and during carbogen breathing for 5 consecutive days. RESULTS: Intracerebral 9L gliomas had a pO(2) of 30-32 mm Hg and C6 gliomas were relatively hypoxic, with a pO(2) of 12-14 mm Hg (p < 0.05). The tissue pO(2) of the contralateral brain was 40-45 mm Hg in rats with either 9L or C6 gliomas. Irradiation resulted in a significant increase in pO(2) of the 9L gliomas only. A significant increase in the pO(2) of the 9L and C6 gliomas was observed in rats breathing carbogen, but this effect decreased during 5 days of repeated experiments in the 9L gliomas. CONCLUSION: These results highlight the tumor specific effect of radiation (9.3.Gy) on tissue pO(2) and the different responses to carbogen inhalation. The ability of electron paramagnetic resonance oximetry to provide direct repeated measurements of tissue pO(2) could have a vital role in understanding the dynamics of hypoxia during therapy that could then be optimized by scheduling doses at times of improved tumor oxygenation. PMID- 19136223 TI - An indirect sandwich ELISA for the detection of avian influenza H5 subtype viruses using anti-hemagglutinin protein monoclonal antibody. AB - A sandwich ELISA test using AIV H5 subtype specific monoclonal antibody (clone 2H4) to an epitope of hemagglutinin protein has been developed. The monoclonal antibody was used to capture the antigen from clinical samples (swabs and tissues). Captured antigens from clinical samples were detected using polyclonal sera, purified AIV H5N1 particles were titrated in the sandwich ELISA and the limit of detection was determined to be approximately 1.0 ng of influenza viral protein in virus preparations. Fifteen AIV strains of H1-H15 subtypes and some other pathogens were tested by this system, and the test is specific to H5 subtype viruses as it failed to detect other AIV subtype viruses and other pathogens. Varieties of clinical samples originating from laboratory experiments (n=382) and from fields (n=288) were employed to test the efficacy of DAS-ELISA test. The test compared very well with the traditional method for detection of influenza virus: virus isolation (VI) in embryonated chicken eggs. In comparison to virus isolation the sensitivity and specificity of sandwich ELISA were found to be 98.6% and 97.6% respectively. In addition, the DAS-ELISA was used to test samples of experimentally infected birds and clinical samples obtained from central China in 2005. The assay proved to be sensitive and specific for the rapid detection of AIV H5 subtype virus form the tissues and swabs in infected animals. PMID- 19136224 TI - Evidence of widespread infection of avian hepatitis E virus (avian HEV) in chickens from Spain. AB - In the present work, 262 serum samples and 29 faeces pools from chickens coming from 29 healthy flocks were analysed by RT-PCR for detection of avian HEV and by ELISA using an aHEV derived antigen for detection of anti-HEV IgG. Additionally, other 300 randomly selected serum samples were also analysed by RT-PCR. Seven serum samples were positive to RNA detection. Sequence analysis of both the helicase and the capsid genes revealed that the Spanish isolates were clustered together and close related to those strains from the United States isolated from farms with HSS. On the serology study, 26/29 flocks had at least one positive animal (89.7%) and chickens older than 40 weeks were found to have higher seropositivities compared to the rest of age groups. Within positive farms, the proportion of positive animals ranged from 20% to 80%. This is the first report of aHEV sequences in chickens from Europe. Further studies are needed to elucidate the clinical significance of avian HEV infections in Europe. PMID- 19136225 TI - Neonatal brain injury: imaging, monitoring and protection. PMID- 19136226 TI - Porcine spermatozoa inhibit post-breeding cytokine induction in uterine epithelial cells in vivo. AB - Early endometrial cytokine responses after exposure to various inseminate components were investigated for a better understanding of the immunological reactions occurring in the porcine uterus after insemination. Baseline values were established for the mRNA concentrations of GM-CSF, IL-6, IL-10, CXCL8 (interleukin-8), Tumour Necrosis Factor alpha (TNF-alpha), TGF-beta, cyclooxygenase-2 (COX-2) and arachidonate 5-lipooxygenase (ALOX-5) in periovulatory uterine endometrial tissue using quantitative RT-PCR. Synchronized gilts were inseminated with spermatozoa diluted either in the semen extender Androhep or seminal plasma. Uterine infusions of media without spermatozoa were used as controls. Three hours after insemination sows were slaughtered and the expression of the above mentioned cytokines was measured in uterine epithelial cells. Simultaneously, the influx of polymorphonuclear neutrophilic (PMN) granulocytes into the uterus was quantified. Compared to baseline values seminal plasma (SP) and Androhep (AH) respectively, if used alone, caused a significant increase in mRNA concentrations of IL-10 (SP: 1.5-fold), TGF-beta (AH: 1.5-fold), CXCL8 (AH: 7.1-fold), TNF-alpha (AH: 1.9-fold) and COX-2 (AH: 7-fold). Surprisingly, in the presence of spermatozoa, none of the tested cytokines revealed mRNA concentrations higher than baseline values. The number of immigrated, intra-luminal PMN correlated only with mRNA concentrations of CXCL8 in presence of Androhep (r=0.51). None of the other cytokines tested seemed to be involved in the regulation of neutrophil recruitment. However, the most interesting result was the sperm-induced down-regulation in the expression of TNF alpha, TGF-beta, IL-10, CXCL8 and COX-2 to mRNA concentration levels similar to or even below baseline values. In conclusion the results show that CXCL8 contributes significantly to uterine PMN recruitment and indicate a so far underestimated role of porcine spermatozoa in the general regulation of the uterine post-mating inflammatory response. PMID- 19136227 TI - Activity-based assessment of the sleep behaviors of VLBW preterm infants and full term infants at around 12 months of age. AB - PURPOSE: The aim of this study is to identify the characteristics of sleep behaviors assessed from activity data among very low-birth weight (VLBW) preterm infants in comparison to full-term infants at the age of around 12 corrected months, due to the possibility that sleep behaviors of preterm infants may be different from those of full-term infants. METHOD: Cross-sectional study. Infant sleep measures were obtained through actigraphs (Micro-mini RC, Ambulatory Monitoring Inc., Ardsley, NY), which were attached to the infants' ankles during a week-long period. The subjects were 14 preterm infants and 14 full-term infants in which the mean ages were 13.3+/-2.1 corrected months and 12.3+/-2.4 month, respectively. RESULTS: Preterm infants showed significantly less sleep duration during nighttime than full-term infants. The mean activity score and ACTX (Activity Index: percentage minutes with more than 0 activity score) of preterm infants during nighttime were significantly higher than those of full-term infants. No significant differences in total sleep duration, daytime sleep duration, the number of night-wakings, and sleep efficiency were found between the two groups. CONCLUSION: Preterm infants slept less and had a larger percentage of less restful sleep during nighttime. The results indicated that prematurity at the time of birth remained throughout the developmental course of sleep behaviors among preterm infants at around the age of 12 corrected months. PMID- 19136228 TI - Patients' trust in their physician--psychometric properties of the Dutch version of the "Wake Forest Physician Trust Scale". AB - OBJECTIVE: Aim was to investigate the psychometric properties of a Dutch version of the "Wake Forest Physician Trust Scale", which intends to measure patients' trust in their physician. METHODS: A random sample of internal medicine patients visiting the outpatient clinic completed the questionnaire (N=201). Dimensionality, reliability and validity of the instrument were examined. RESULTS: The structure of the questionnaire was best explained by a unidimensional construct. Reliability was confirmed: internal consistency was high (alpha=.88), and mean item-total correlations were all above .40. Construct validity was indicated by patients' trust in their physician correlating significantly and as hypothesized with (1) satisfaction with their physician (r=.64), (2) with the length of the patient-physician relationship (r=.28), (3) with their willingness to recommend their physician (r=.71) and (4) their unwillingness to switch their physician (r=.61). CONCLUSION: The results suggest the Dutch version of the Wake Forest Physician Trust Scale to be a psychometrically sound instrument to assess patients' interpersonal trust. PRACTICE IMPLICATIONS: Trust is a key feature of the patient-physician relationship, yet has been scarcely researched in other than Anglophone cultures. An adequate Dutch trust questionnaire forms the first step to gaining more knowledge about patient-physician trust in another culture and health care setting. PMID- 19136229 TI - A practitioner's guide to persuasion: an overview of 15 selected persuasion theories, models and frameworks. AB - OBJECTIVE: To provide a brief overview of 15 selected persuasion theories and models, and to present examples of their use in health communication research. RESULTS: The theories are categorized as message effects models, attitude behavior approaches, cognitive processing theories and models, consistency theories, inoculation theory, and functional approaches. CONCLUSIONS: As it is often the intent of a practitioner to shape, reinforce, or change a patient's behavior, familiarity with theories of persuasion may lead to the development of novel communication approaches with existing patients. PRACTICE IMPLICATIONS: This article serves as an introductory primer to theories of persuasion with applications to health communication research. Understanding key constructs and general formulations of persuasive theories may allow practitioners to employ useful theoretical frameworks when interacting with patients. PMID- 19136230 TI - [How can we use antihyperalgesic drugs?]. PMID- 19136231 TI - Activated protein C resistance in deep venous thrombosis. AB - Several risk factors for deep venous thrombosis (DVT) have been identified, and inherited thrombophilias constitute a significant proportion of them. The most common inherited thrombophilia is activated protein C (APC) resistance, and factor V Leiden is the most common cause of APC resistance. The high prevalence of APC resistance in Caucasians is established, and the prevalence among persons of Asian and African descent has been shown to be low in previous studies. Twenty three patients with lower limb DVT were included in the study. Diagnosis was confirmed by duplex ultrasonography. Patients were tested for APC resistance with use of the STA Staclot APC-R system (Diagnostica Stago, Asnieres, France), as per the manufacturer's guidelines. Ten patients (43.5%) tested positive for APC resistance, while 13 (56.5%) tested negative. APC resistance, although considered a rarity, has been found to have a high prevalence in patients with DVT from the northeastern region of India. APC resistance estimation should be done for all patients with DVT. PMID- 19136233 TI - Updating meta-analyses leads to larger type I errors than publication bias. AB - OBJECTIVE: To estimate the extent to which the practice of periodically updating meta-analyses causes inflation of the type I error and then to compare the estimate with the inflation caused by publication bias. We also present a simple method to adjust for the inflation associated with updating meta-analyses. STUDY DESIGN AND SETTING: Simulations were used to estimate the error rates. RESULTS: In general, updating meta-analyses caused 2- to 5-fold inflation of the type I error rates, which exceeded the inflation caused by publication bias. As a rule of thumb, the results of a meta-analysis are robust up to 5, 10, 15, or 22 updates, if the P-value multiplied by 4, 6, 8, or 10 remains below the desired significance level. CONCLUSION: Meta-analyses are likely to be updated until a clear conclusion is reached. Therefore, it is important to take the inflation of the error rate into account to interpret the results correctly. PMID- 19136232 TI - Outcome and survival of patients aged 75 years and older compared to younger patients after ruptured abdominal aortic aneurysm repair: do the results justify the effort? AB - We evaluated early mortality (<30 days) rates, cost analyses, and preoperative variables that may be predictive of 30-day mortality in elderly patients compared to younger patients after emergency open repair of ruptured abdominal aortic aneurysm (RAAA). The survey is a retrospective analysis based on prospectively registered data. The protocol was an "all-comers" policy. Seventy-two patients, who were operated on for RAAA in our department from January 1, 2005, to December 30, 2005, are included in this study. The follow-up time of survivors was 1 year. We defined 75-year-old patients as elderly because of the increased incidence of surgical risk factors and hospital mortality in this subset of patients (cut-off age). Demographic, clinical, and operative factors were analyzed together with 30 day mortality. Univariate analysis was performed with the chi-squared test. Multivariate analyses were also performed with the variables that were found to be significant in the univariate analysis. Health economy and cost analysis for the two groups were estimated. Out of 72 open repairs of RAAA, 44 patients (61%) were under 75 years of age and 28 (39%) were 75 years or older. The average age of the patients was 71 years (confidence interval [CI] 69.2-73.7, range 53-87). Twenty-five patients (35%, CI 27.6-51.2) died within 30 days in the postoperative period. The 30-day mortality for the 28 elderly patients who underwent open operative repair was 16 (57%, CI 48%-72%) compared to 9 (20%, CI 12%-33%) of 44 younger patients (p < 0.001). An age of 75 years or older and a serum creatinine >or=0.150 mmol/L in elderly patients with RAAA (p < 0.01) were identified to be significant risk factors for operative mortality. We did not encounter significant differences in the distribution of other risk factors in the group of elderly patients compared to the younger group. Between the survivors of the two groups, there were no significant differences in the total length of stay (LOS) and the LOS in the intensive care unit. Advanced age (>or=75) and the combination of this advanced age and serum creatinine of >or=0.150 mmol/L were the only significant (p < 0.05) preoperative risk factors in our single-center study. However, we believe that treatment for RAAA can be justified in elderly patients. In our experience, surgical open repair has been life-saving in 33% of patients aged 75 years and older, at a relatively low price for each life, estimated at euro 40,409. PMID- 19136234 TI - Systematic reviews of low back pain prognosis had variable methods and results: guidance for future prognosis reviews. AB - OBJECTIVE: Systematic reviews of prognostic factors for low back pain vary substantially in design and conduct. The objective of this study was to identify, describe, and synthesize systematic reviews of low back pain prognosis, and explore the potential impact of review methods on the conclusions. STUDY DESIGN AND SETTING: We identified 17 low back pain prognosis reviews published between 2000 and 2006. One reviewer extracted and a second checked review characteristics and results. Two reviewers independently assessed review quality. RESULTS: Review questions and selection criteria varied; there were both focused and broad reviews of prognostic factors. A quarter of reviews did not clearly define search strategies. The number of potential citations identified ranged from 15 to 4,458 and the number of included prognosis studies ranged from 3 to 32 (of 162 distinct citations included across reviews). Seventy percent of reviews assessed quality of included studies, but assessed only a median of four of six potential biases. All reviews reported associations based on statistical significance; they used various strategies for syntheses. Only a small number of important prognostic factors were consistently reported: older age, poor general health, increased psychological or psychosocial stress, poor relations with colleagues, physically heavy work, worse baseline functional disability, sciatica, and the presence of compensation. We found discrepancies across reviews: differences in some selection criteria influenced studies included, and various approaches to data interpretation influenced review conclusions about evidence for specific prognostic factors. CONCLUSION: There is an immediate need for methodological work in the area of prognosis systematic reviews. Because of methodological shortcomings in the primary and review literature, there remains uncertainty about reliability of conclusions regarding prognostic factors for low back pain. PMID- 19136235 TI - Seizure-like activity in the hypoglycemic rat: lack of correlation with the electroencephalogram of free-moving animals. AB - BACKGROUND: The neuropathology of hypoglycemia and its mechanisms have been well studied. However, the physiopathogenesis of hypoglycemia-related seizures has escaped elucidation. Various animal models reportedly show "seizures" when rendered hypoglycemic, however, correlation with the electroencephalogram (EEG) is inconsistent. In order to characterize the role of the hippocampus and frontal neocortex in the generation of hypoglycemic seizures, this study was undertaken. METHODS: Adult rats were implanted stereotaxically with electrodes in the left hippocampus and right frontal cortex. After 1 week, they were fasted 18-24h, then injected intraperitoneally with insulin, 35 IU/kg. Simultaneous EEG/video monitoring was conducted. RESULTS: Interpretable EEG recordings were obtained in 8/12 animals. Two showed poor association of seizure-like behaviour (neck extension, vocalizations, tonic extension of the tail, digging or running limb movements) with ictal EEG patterns. Four animals exhibited such behaviours during periods of high amplitude polymorphic slow wave activity, burst-suppression patterns or non-rhythmic spiking. Two others were encephalopathic (behaviourally and electroencephalographically) until death. CONCLUSIONS: Not all animals develop seizure-like behaviour when hypoglycemic. If these are seizures, they may originate from subcortical structures, or the "convulsive" behaviours observed may simply be flight/fight reflexes released during profound encephalopathy. Spike activity in the EEG may be a manifestation of this state. Recording EEG from rat cortex and hippocampus during seizure-like activity brought on by hypoglycemia correlates poorly with seizure-like behaviours suggesting that the relevant electrophysiological correlates, if present, are generated from deeper brain structures. PMID- 19136236 TI - Clinical, histological and genetic investigation of Buschke-Fischer-Brauer's disease in Tunisian families. PMID- 19136237 TI - The effects of diethyldithiocarbamate, a SOD inhibitor, on endothelial function in sedentary and exercised db/db mice. AB - The study was designed to evaluate the effects of diethyldithiocarbamate (DDC), an superoxide dismutase (SOD) blocker on endothelial function in db/db mice. The db/db and wild-type (WT) mice were randomly divided into low intensity exercise, moderate intensity exercise and control (sedentary) groups. Mice were exercised daily, 5 days per week. After 6 weeks, ring segments of aortae were mounted in wire myograph and acetylcholine (ACh) concentration response curves were recorded in absence or presence of DDC and DDC plus SOD. Results showed that ACh concentration response curve was similar in WT and WT exercised groups. Incubation of aorta rings of WT mice with DDC suppressed the maximum ACh response (p<0.05). Subsequent incubation with SOD restored vasodilatory response in WT mice. The vasodilatation to ACh was significantly reduced in sedentary db/db mice compare to WT (p<0.05) and incubation with DDC did not further decrease this response, however, addition of SOD restored the vasodilatation to ACh to that observed in WT mice. Mild and moderate exercised db/db mice had ACh response similar to that in WT mice. Incubation with DDC incubation reduced ACh induced vasodilatation and addition of SOD restored this response. Our results support the conclusion that SOD mimetics can be used to improve superoxide-mediated endothelial dysfunction in diabetic db/db mice. PMID- 19136238 TI - Co-morbid association of depression and COPD: a population-based study. AB - OBJECTIVES: Depression occurs commonly among patients with COPD, but the independent association of depression and COPD and the effect of depression on COPD outcomes are not well established. METHOD: A population sample of 2402 Chinese aged >or=55 with and without COPD (characteristic symptoms of chronic cough, sputum or breathlessness and airflow obstruction and FEV(1)/FVC<0.70) was assessed on Geriatric Depression Scale (score>or=5), dependence on basic activities of daily living (ADL), SF-12 health status, smoking and medication behaviour. RESULTS: The 189 respondents with COPD showed higher depressive symptoms prevalence (22.8%) than 2213 respondents without COPD (12.4%); multivariate odd ratio (OR) was 1.86; 95% CI, 1.25-2.75 after controlling for confounding risk factors. In multivariate analyses of respondents with COPD, those who were depressed (N=43), compared to those who were not (N=146), were more likely to report ADL disability (OR=2.89, p=0.049) poor or fair self reported health (OR=3.35, p=0.004), poor SF-12 PCS scores (OR=2.35, p=0.041) and SF-12 MCS scores (OR=4.17, p<0.001). CONCLUSION: Depressive symptoms were associated with COPD independent of known risk factors. In COPD participants, depressive symptoms were associated with worse health and functional status and self-management. PMID- 19136239 TI - Potential side effects in patients treated with inhaled corticosteroids and long acting beta2-agonists. AB - BACKGROUND: Long-acting-beta-agonists and inhaled corticosteroids are widely used in patients with obstructive lung disease. We determined the occurrence of potential side effects to inhaled steroids and long-acting-beta-agonists in an open post-marketing observational study. METHODS: A total of 158 adult patients treated with long-acting-beta-agonists and inhaled steroids because of asthma or chronic obstructive lung disease were included prospectively in a cross-sectional study from September 2004 through august 2005. A subgroup of 31 patients with minimal disease were observed prospectively after treatment reduction. The frequency and intensity of potential side effects to long-acting-beta-agonists and inhaled corticosteroids were registered on a 10-point visual analogue scale. RESULTS: A total of 131 (83%) reported potential side effects to inhaled steroids. The most frequent were sore (54%) and dry (52%) throat. A total of 114 (72%) patients reported potential side effects to long-acting-beta-agonists. The most frequent were muscle cramps (62%) and muscle twisting (39%). For 5/8 potential side effects their occurrence increased with increased individual dozing. In 31 patients the mean medication with steroids and long-acting-beta agonists was reduced by 87% and 97.5%, respectively, with side effects reduced by 62% (p<0.001) to inhaled steroids and by 91% (p<0.001) to inhaled long-acting beta-agonists. CONCLUSIONS: Potential side effects in adult patients to inhaled corticosteroids and long-acting-beta-agonists are very common with increased frequency with increased dozing. Patients are largely unaware of the association. Patients should be better informed, and the results further support tailored dozing to minimum therapy. PMID- 19136240 TI - Medication adherence and persistence in the last year of life in COPD patients. AB - OBJECTIVE: To examine medication adherence and persistence among COPD patients during their last year of life. DATA SOURCE: National VA databases were used to identify patients who had COPD and died between 1999 and 2003. STUDY DESIGN: We examined use of inhaled corticosteroids (ICS), long acting beta(2) agonists (LABA), methylxanthines (MTX), and anticholinergics (AC), alone and in combination. Medication possession ratios (MPR) were compared between regimens by quarterly periods using General Estimating Equations (GEE). Medication persistence was examined in monotherapy users with Kaplan-Meier survival analysis and extended Cox proportional hazard models. PRINCIPAL FINDINGS: Only half of the identified patients in the COPD cohort (5913 of 11,376) used any medications. Among 5913 patients, overall mean (SD) MPR was 0.44 (0.32) during the last year of life. A positive linear trend in MPR was observed across quarterly periods in AC users (beta=0.014, p<0.0001), and was highest for MTX users (beta=0.11, p<0.0001). Of 3436 on monotherapy only, 40% discontinued medication within 30 days, and 70% discontinued within 90 days. MTX users were less likely to discontinue (HR=0.714, p=0.012) than reference (AC) group. CONCLUSION: COPD patients in their last year of life tended to use respiratory medications sporadically. Further research is needed to qualify whether minor differences in MPR between regimens reflect behavioral differences related to regimen or reflect refill policy and MPR calculation technique. PMID- 19136241 TI - Capsaicin cough sensitivity in smokers with and without airflow obstruction. AB - BACKGROUND: Cough is a frequent symptom of cigarette smokers that often precedes the development of airflow obstruction. We determined whether chronic cigarette smoking is associated with an increase in capsaicin cough response in the absence of cough. METHODS: We examined this in asymptomatic smokers with normal lung function (n=68, FEV(1) 99.3+/-2.1% predicted) and in patients with established COPD without cough symptoms (n=42; FEV(1) 57.0+/-2.6% predicted), using healthy non-smoking volunteers as control (n=92; FEV(1) 100.6+/-1.7% predicted). Using an incremental capsaicin concentration challenge protocol, we recorded the concentrations that induced 2 (C2) and 5 or more coughs (C5). RESULTS: Because females have a lower C2 and C5 than males in the control group, we analysed the data in each group according to gender. Log C5 was decreased both in asymptomatic smokers (1.56+/-0.11 micromol/L, p<0.05) and in COPD patients (1.44+/-0.14 micromol/L, p<0.01) when compared to non-smokers (1.90+/-0.09 micromol/L). Log C2 did not differ between groups. Log C2 and log C5 were decreased in women (0.772+/ 0.071 micromol/L and 1.481+/-0.094 micromol/L, respectively) when compared to men (1.045+/-0.088 micromol/L and 1.923+/-0.087 micromol/L, respectively) (p<0.05 for log C2; p<0.001 for log C5). CONCLUSION: We conclude that chronic cigarette smoking increases capsaicin cough reflex and that this remains so with the development of COPD. PMID- 19136242 TI - Analysis of potential RDF resources from solid waste and their energy values in the largest industrial city of Korea. AB - The production potential of refuse derived fuel (RDF) in the largest industrial city of Korea is discussed. The purpose of this study is to evaluate the energy potential of the RDF obtained from utilizing combustible solid waste as a fuel resource. The total amount of generated solid waste in the industrial city was more than 3.3 million tonnes, which is equivalent to 3.0tonnes per capita in a single year. The highest amount of solid waste was generated in the city district with the largest population and the biggest petrochemical industrial complex (IC) in Korea. Industrial waste accounted for 89% of the total amount of the solid waste in the city. Potential RDF resources based on combustible solid wastes including wastepaper, wood, rubber, plastic, synthetic resins and industrial sludge were identified. The amount of combustible solid waste that can be used to produce RDF was 635,552tonnes/yr, consisting of three types of RDF: 116,083tonnes/yr of RDF-MS (RDF from municipal solid waste); 146,621tonnes/yr of RDF-IMC (RDF from industrial, municipal and construction wastes); and 372,848tonnes/yr of RDF-IS (RDF from industrial sludge). The total obtainable energy value from the RDF resources in the industrial city was more than 2,240,000x10(6)kcal/yr, with the following proportions: RDF-MS of 25.6%, RDF-IMC of 43.5%, and RDF-IS of 30.9%. If 50% or 100% of the RDF resources are utilized as fuel resources, the industrial city can save approximately 17.6% and 35.2%, respectively, of the current total disposal costs. PMID- 19136243 TI - Life cycle assessment of bagasse waste management options. AB - Bagasse is mostly utilized for steam and power production for domestic sugar mills. There have been a number of alternatives that could well be applied to manage bagasse, such as pulp production, conversion to biogas and electricity production. The selection of proper alternatives depends significantly on the appropriateness of the technology both from the technical and the environmental points of view. This work proposes a simple model based on the application of life cycle assessment (LCA) to evaluate the environmental impacts of various alternatives for dealing with bagasse waste. The environmental aspects of concern included global warming potential, acidification potential, eutrophication potential and photochemical oxidant creation. Four waste management scenarios for bagasse were evaluated: landfilling with utilization of landfill gas, anaerobic digestion with biogas production, incineration for power generation, and pulp production. In landfills, environmental impacts depended significantly on the biogas collection efficiency, whereas incineration of bagasse to electricity in the power plant showed better environmental performance than that of conventional low biogas collection efficiency landfills. Anaerobic digestion of bagasse in a control biogas reactor was superior to the other two energy generation options in all environmental aspects. Although the use of bagasse in pulp mills created relatively high environmental burdens, the results from the LCA revealed that other stages of the life cycle produced relatively small impacts and that this option might be the most environmentally benign alternative. PMID- 19136244 TI - Thermal processing of paper sludge and characterisation of its pyrolysis products. AB - Paper sludge is a waste product from the paper and pulp manufacturing industry that is generally disposed of in landfills. Pyrolysis of paper sludge can potentially provide an option for managing this waste by thermal conversion to higher calorific value fuels, bio-gas, bio-oils and charcoal. This work investigates the properties of paper sludge during pyrolysis and energy required to perform thermal conversion. The products of paper sludge pyrolysis were also investigated to determine their properties and potential energy value. The dominant volatile species of paper sludge pyrolysis at 10 degrees C/min were found to be CO and CO(2), contributing to almost 25% of the paper sludge dry weight loss at 500 degrees C. The hydrocarbons (CH(4), C(2)H(4), C(2)H(6)) and hydrogen contributed to only 1% of the total weight loss. The bio-oils collected at 500 degrees C were primarily comprised of organic acids with the major contribution being linoleic acid, 2,4-decadienal acid and oleic acid. The high acidic content indicates that in order to convert the paper sludge bio-oil to bio diesel or petrochemicals, further upgrading would be necessary. The charcoal produced at 500 degrees C had a calorific value of 13.3MJ/kg. PMID- 19136245 TI - Geographic information systems (GIS) based model of dairy manure transportation and application with environmental quality consideration. AB - Survey information was used to develop a minimum cost spatial dairy manure transportation model where environmental quality and crop nutrient requirements were treated as constraints. The GIS model incorporated land use types, exact locations of dairy farms and farmlands, road networks, and distances from each dairy farm to receiving farmlands to identify dairy manure transportation routes that minimized costs relative to environmental and other constraints. Our analyses indicated that the characteristics of dairy manure, its bulk and relatively low primary N, P(2)O(5) and K(2)O nutrient levels limit the distribution areas or distances between the farms and the land over which the manure can be economically spread. Physical properties of the land limit the quantities of nutrients that can be applied because of excess nutrient buildup in soil and potential to harm nearby waterbodies and downstream people and places. Longer distances between dairy and farmland favor the use of commercial fertilizers due to the high cost of manure transportation. At $0.08 per ton per km transportation cost, the optimal cut-off distances for dairy manure application is 30km for N and 15km each for P(2)O(5) and K(2)O consistent rules. An analysis of dairy manure application to different crop types suggest that, on average, 1ha of land requires 61 tons of dairy manure to meet the recommended N, P(2)O(5) and K(2)O needs. PMID- 19136246 TI - Application of risk management techniques for the remediation of an old mining site in Greece. AB - This article summarizes the project and risk management of a remediation/reclamation project in Lavrion, Greece. In Thoricos the disposal of mining and metallurgical wastes in the past resulted in the contamination with heavy metals and acid mine drainage. The objective of this reclamation project was to transform this coastal zone from a contaminated site to an area suitable for recreation purposes. A separate risk assessment study was performed to provide the basis of determining the relevant environmental contamination and to rate the alternative remedial schemes involved. The study used both existing data available from comprehensive studies, as well as newly collected field data. For considering environmental risk, the isolation and minimization of risk option was selected, and a reclamation scheme, based on environmental criteria, was applied which was comprised of in situ neutralization, stabilization and cover of the potentially acid generating wastes and contaminated soils with a low permeability geochemical barrier. Additional measures were specifically applied in the areas where highly sulphidic wastes existed constituting active acid generation sources, which included the encapsulation of wastes in HDPE liners installed on clay layers. PMID- 19136247 TI - Removal of copper, chromium and arsenic from preservative-treated wood by chemical extraction-fungal bioleaching. AB - Large volumes of preservative-treated wood containing toxic Cr, Cu and As salts are decommissioned worldwide. This study investigated the effectiveness of solid state fermentation with copper-tolerant brown-rot fungi for the remediation of wood treated with chromated copper arsenate (CCA) and acid copper chromate (ACC) formulations. Treatment of CCA- and ACC-wood with the most effective strain, Antrodia vaillantii FRLP-14G, attained extensive leaching of As and/or Cr, but Cu elimination was poor (<18%). Additional research showed that a variety of organic acids, including citrate, are effective Cu extractants. Based on these findings, a process combining chemical extraction and subsequent fungal treatment was developed that proved highly effective in removing inorganic pollutants from CCA wood. Extraction of CCA-wood with citric acid (30 mM, pH 3.10) followed by a 28 day solid-state fermentation period removed 87% Cu, 80% Cr, and 100% As. These results indicate the potential of the two-stage process for the remediation of preservative-treated wood. PMID- 19136248 TI - Biochip functionalization using electrowetting-on-dielectric digital microfluidics for surface plasmon resonance imaging detection of DNA hybridization. AB - This work reports on a dynamically configurable micro-array surface plasmon resonance biochip platform. The platform comprises a digital electrowetting-on dielectric (EWOD) microfluidic device tailored to surface plasmon resonance imaging (SPRi). We demonstrate its application for simultaneous immobilization of different DNA probes at the designated detection sites on-chip from sub-microL volume solutions in combination with multichannel label-free real-time detection of subsequent hybridization reactions. Successful on-chip DNA probe dilution and immobilization is also demonstrated using SPRi hybridization detection. Furthermore, active control of the immobilized probe density and orientation is achieved under an applied potential using the electric interface of the EWOD device. For low probe densities, under negative applied potential, the DNA hybridization efficiency is enhanced compared to passive probe immobilization, yielding a two-fold SPR signal increase within only 8min of hybridization. EWOD microfluidic platform coupled with SPRi promises to dramatically increase the speed of detection and quantification of biomolecular interactions while reducing reagent consumption. The proposed system would enable the development of high throughput, rapid and ultrasensitive detection of biomolecules beyond DNA microarray applications. PMID- 19136249 TI - Preclinical evaluation of vascular-disrupting agents in Ewing's sarcoma family of tumours. AB - The effects of the tubulin-binding vascular-disrupting agents (VDAs), combretastatin A4 phosphate (CA4P), OXi4503/CA1P and OXi8007, in subcutaneous mouse models of the Ewing's sarcoma family of tumours (ESFTs) have been investigated alone and in combination with doxorubicin. Delay in subcutaneous tumour growth was observed following treatment of mice with multiple doses of OXi4503/CA1P but not with CA4P or OXi8007. A single dose of OXi4503/CA1P caused complete shutdown of vasculature by 24h and extensive haemorrhagic necrosis by 48h. However, a viable rim of proliferating cells remained, which repopulated the tumour within 10 days following the withdrawal of treatment. Combined treatment with doxorubicin 1h prior to administration of OXi4503/CA1P enhanced the effects of OXi4503/CA1P causing a synergistic delay in tumour growth (p<0.001). This study demonstrates that OXi4503/CA1P is a potent VDA in ESFT and in combination with conventional cytotoxic agents represents a promising treatment strategy for this tumour group. PMID- 19136250 TI - Interleukin 7/interleukin 7 receptor induce c-Fos/c-Jun-dependent vascular endothelial growth factor-D up-regulation: a mechanism of lymphangiogenesis in lung cancer. AB - Interleukin 7 (IL-7) is known to promote lymphangiogenesis. To study the relationship between IL-7 and the lymphangiogenic factor, vascular endothelial growth factor (VEGF)-D, in human lung cancer cells and its impact on the prognosis of lung cancer patients, we investigated how IL-7 regulates VEGF-D. We found that, in lung cancer cell lines, IL-7/IL-7 receptor (IL-7R) increase the expression of VEGF-D and phosphorylation of c-Fos/c-Jun, induce c-Fos and c-Jun heterodimer formation, and enhance c-Fos/c-Jun DNA binding activity to regulate VEGF-D. In addition, the expression levels of IL-7 and IL-7R correlated well with that of VEGF-D, lymphatic vessels density (LVD), clinical stages, lymph node metastasis, and poor prognosis in 100 human non-small cell lung cancer (NSCLC) specimens analysed. Taken together, our results provide evidence that IL-7/IL-7R induce VEGF-D up-regulation and promote lymphangiogenesis via c-Fos/c-Jun pathway in lung cancer. PMID- 19136251 TI - The impact of socioeconomic factors on 30-day mortality following elective colorectal cancer surgery: a nationwide study. AB - We investigated postoperative mortality in relation to socioeconomic status (SES) in electively operated colorectal cancer patients, and evaluated whether social inequalities were explained by factors related to patient, disease or treatment. Data from the nationwide database of Danish Colorectal Cancer Group were linked to individual socioeconomic information in Statistics Denmark. Patients born before 1921 and those having local surgical or palliative procedures were excluded. A total of 7160 patients, operated on in the period 2001-2004, were included, of whom 342 (4.8%) died within 30 days of surgery. Postoperative mortality was significantly lower in patients with high income (odds ratio (OR)=0.82 (0.70-0.95) for each increase in annual income of EUR 13,500), higher education versus short education (OR)=0.60 (0.41-0.87), and owner-occupied versus rental housing (OR)=0.73 (0.58-0.93). Differences in comorbidity and to a lesser extent lifestyle characteristics accounted for the excess risk of postoperative death among low-SES patients. PMID- 19136252 TI - Investigation of the microbial community structure and activity as indicators of compost stability and composting process evolution. AB - In a bid to identify suitable microbial indicators of compost stability, the process evolution during windrow composting of poultry manure (PM), green waste (GW) and biowaste was studied. Treatments were monitored with regard to abiotic factors, respiration activity (determined using the SOUR test) and functional microflora. The composting process went through typical changes in temperature, moisture content and microbial properties, despite the inherent feedstock differences. Nitrobacter and pathogen indicators varied as a monotonous function of processing time. Some microbial groups have shown a potential to serve as fingerprints of the different process stages, but still they should be examined in context with respirometric tests and abiotic parameters. Respiration activity reflected well the process stage, verifying the value of respirometric tests to access compost stability. SOUR values below 1 mg O(2)/g VS/h were achieved for the PM and the GW compost. PMID- 19136253 TI - The investigation of effect of organic carbon sources addition in anaerobic aerobic (low dissolved oxygen) sequencing batch reactor for nutrients removal from wastewaters. AB - The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic-aerobic (low dissolved oxygen, 0.15-0.45 mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations. PMID- 19136254 TI - Ecological significance and some biotechnological application of an organic solvent stable alkaline serine protease from Bacillus subtilis strain DM-04. AB - An organic solvent stable, alkaline serine protease (Bsubap-I) with molecular mass of 33.1 kDa, purified from Bacillus subtilis DM-04 showed optimum activity at temperature and pH range of 37-45 degrees C and 10.0-10.5, respectively. The enzyme activity of Bsubap-I was significantly enhanced in presence of Fe(2+). The thermal resistance and stability and of Bsubap-I in presence of surfactants, detergents, and organic solvents, and its dehairing activity supported its candidature for application in laundry detergent formulations, ultrafiltration membrane cleaning, peptide synthesis and in leather industry. The broad substrate specificity and differential antibacterial property of Bsubap-I suggested the natural ecological role of this enzyme for the producing bacterium. PMID- 19136255 TI - Characterization of bio-oil from induction-heating pyrolysis of food-processing sewage sludges using chromatographic analysis. AB - In this study, gas chromatography-mass spectrometry (GC-MS) was used to analyze the pyrolytic bio-oils and gas fractions derived from the pyrolysis of industrial sewage sludges using induction-heating technique. The liquid products were obtained from the cryogenic condensation of the devolatilization fraction in a nitrogen atmosphere using a heating rate of 300 degrees C/min ranging from 25 to 500 degrees C. The analytical results showed that the pyrolysis bio-oils were very complex mixtures of organic compounds and contained a lot of nitrogenated and/or oxygenated compounds such as aliphatic hydrocarbons, phenols, pyridines, pyrroles, amines, ketones, and so on. These organic hydrocarbons containing nitrogen and/or oxygen should originate from the protein and nucleic acid textures of the microbial organisms present in the sewage sludge. The non condensable devolatilization fractions were also composed of nitrogenated and oxygenated compounds, but contained small fractions of phenols, 1H-indoles, and fatty carboxylic acids. On the other hand, the compositions in the non condensable gas products were principally carbon dioxide, carbon monoxide and methane analyzed by gas chromatography-thermal conductivity detector (GC-TCD). PMID- 19136256 TI - Multiscale modelling of hydrothermal biomass pretreatment for chip size optimization. AB - The objective of this work is to develop a relationship between biomass chip size and the energy requirement of hydrothermal pretreatment processes using a multiscale modelling approach. The severity factor or modified severity factor is currently used to characterize some hydrothermal pretreatment methods. Although these factors enable an easy comparison of experimental results to facilitate process design and operation, they are not representative of all the factors affecting the efficiency of pretreatment, because processes with the same temperature, residence time, and pH will not have same effect on biomass chips of different size. In our study, a model based on the diffusion of liquid or steam in the biomass that takes into account the interrelationship between chip size and time is developed. With the aid of our developed model, a method to find the optimum chip size that minimizes the energy requirement of grinding and pretreatment processes is proposed. We show that with the proposed optimization method, an average saving equivalent to a 5% improvement in the yield of biomass to ethanol conversion process can be achieved. PMID- 19136257 TI - Discovery of novel small molecule cell type-specific enhancers of NF-kappaB nuclear translocation. AB - An IKKbeta inhibitor reported to block NF-kappaB transcriptional activities in Jurkat T cells, was found to enhance NF-kappaB translocation in HUVEC cells. These studies suggested a noncanonical NF-kappaB signaling pathway independent of IKKbeta in HUVEC cells. PMID- 19136258 TI - Synthesis, biological activity and tubulin binding poses of 1-deoxy-9-(R) dihydrotaxane analogs. AB - 1-Deoxy-9alpha-dihydrotaxane analogs 9 and 10 were semi-synthesized from 1 deoxybaccatin VI, isolated from Taxus mairei, and tested for cytotoxic activity. Taxane 9 is 10-fold less cytotoxic than paclitaxel, while 10 is equally active. In the tubulin polymerization assay (ED(50) values), 10 is 4-fold less effective than paclitaxel, but 3-fold superior to 9. These observations can be explained by analysis of the corresponding taxane/beta-tubulin complexes. PMID- 19136260 TI - Synthesis and evaluation of a cyclic imine derivative conjugated to a fluorescent molecule for labeling of proteins. AB - A cyclic imine conjugated to a fluorescent dansyl group was synthesized and used for covalent labeling of proteins. The covalent attachment to proteins was confirmed by gel electrophoresis and mass analysis. PMID- 19136259 TI - Synthesis of 1-(methanesulfonyl- and aminosulfonylphenyl)acetylenes that possess a 2-(N-difluoromethyl-1,2-dihydropyridin-2-one) pharmacophore: evaluation as dual inhibitors of cyclooxygenases and 5-lipoxygenase with anti-inflammatory activity. AB - A hitherto unknown class of linear acetylene regioisomers were designed such that a SO(2)Me or SO(2)NH(2) group was located at the ortho-, meta- or para-position of the acetylene C-1 phenyl ring, and a N-difluoromethyl-1,2-dihydropyridin-2-one moiety was attached via its C-5 position to the C-2 position on an acetylene template (scaffold). All three SO(2)Me regioisomers, and the 4-SO(2)NH(2) analog, were potent inhibitors of 5-lipoxygenase (5-LOX IC(50)=3.2-3.5 microM range) relative to the reference drug caffeic acid (IC(50)=4.0 microM). The SO(2)Me regioisomers exhibited weak cyclooxygenease-1 (COX-1) and -2 (COX-2) inhibitory activity with a modest COX-2 selectivity index. The most potent 3-SO(2)Me, 4 SO(2)Me and 4-SO(2)NH(2) compounds, with respective ED(50) values of 66.1, 68.5 and 86.5 mg/kg po, exhibited comparable oral anti-inflammatory (AI) activity to that of the reference drug ibuprofen (ED(50)=67.4 mg/kg po). The N-difluoromethyl 1,2-dihydropyridin-2-one moiety provides a novel pharmacophore for the design of cyclic hydroxamic mimetics capable of inhibiting 5-LOX for exploitation in the development of 5-LOX inhibitory AI drugs. PMID- 19136261 TI - Endovascular embolization of a recurrent cervical giant cell neoplasm using N butyl 2-cyanoacrylate. AB - Pre-operative endovascular embolization of spinal giant cell tumors (GCTs) has been an effective strategy to reduce blood loss during surgical resection. Traditionally, spinal GCTs have been embolized with polyvinyl acetate (PVA) particles. We present the pre-operative embolization of a recurrent cervical GCT with N-butyl 2-cyanoacrylate (NBCA) rather than PVA. The patient was a 17-year old female who, 3 months prior, had undergone a surgical resection of a cervical GCT without pre-operative embolization. She returned with tumor recurrence in the approximate location. Resection was recommended, and pre-operative embolization was requested. The tumor was embolized with NBCA. Post-embolization angiography demonstrated significantly decreased tumor "blush" and a significant reduction of the vascular supply. This is the first reported use of NBCA for the pre-operative embolization of a cervical GCT. The benefits of NBCA over PVA particles include superior penetration, permanent tumor embolization and lower exposure to radiation due to shorter procedure time. PMID- 19136262 TI - Glomus tumor and knee pain: a report of four cases. AB - We present four cases of glomus tumors presenting as knee pain. All cases were treated by surgical excision of the tumor. All patients made an immediate recovery with return to full normal function. The presentation of this is unique in that the patient has exquisite pain and tenderness when the area affected is palpated. Occasionally, local infiltration or an ischaemia test can assist with diagnosis. To our knowledge, this is the largest case series in the literature. PMID- 19136263 TI - Synthesis and biological evaluation of extraordinarily potent C-10 carba artemisinin dimers against P. falciparum malaria parasites and HL-60 cancer cells. AB - A series of artemisinin dimers incorporating a metabolically stable C-10 carba linkage have been prepared, several of which show remarkable in vitro antimalarial activity (as low as 30 pM) versus Plasmodium falciparum and in vitro anticancer activity in the micromolar to nanomolar range versus HL-60 cell lines. PMID- 19136264 TI - Development of a rapid source preparation method for high-resolution alpha particle spectrometry. AB - In order to prepare sources of short-lived actinides for alpha-particle spectrometry, a coprecipitation method with Sm hydroxide was developed. The preparation procedure can be completed within 5 min with a high chemical yield of over 90%. It was found that the uniformity of the produced sources was sufficient to provide a high resolution of better than 20 keV. Under this method, we successfully measured the alpha-particle spectrum of short-lived Cf isotopes produced in the (238)U((12)C, xn) reaction. PMID- 19136265 TI - WITHDRAWN: Measurement of beam focus quality in biomedical nuclear microscopy. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19136266 TI - Putrescine overproduction negatively impacts the oxidative state of poplar cells in culture. AB - While polyamines (PAs) have been suggested to protect cells against Reactive Oxygen Species (ROS), their catabolism is known to generate ROS. We compared the activities of several enzymes and cellular metabolites involved in the ROS scavenging pathways in two isogenic cell lines of poplar (Populus nigraxmaximowiczii) differing in their PA contents. Whereas the control cell line was transformed with beta-glucuronidase (GUS), the other, called HP (High Putrescine), was transformed with a mouse ornithine decarboxylase (mODC) gene. The expression of mODC resulted in several-fold increased production of putrescine as well its enhanced catabolism. The two cell lines followed a similar trend of growth over the seven-day culture cycle, but the HP cells had elevated levels of soluble proteins. Accumulation of H(2)O(2) was higher in the HP cells than the control cells, and so were the activities of glutathione reductase and monodehydroascorbate reductase; the activity of ascorbate peroxidase was lower in the former. The contents of reduced glutathione and glutamate were significantly lower in the HP cells but proline was higher on some days of analysis. There was a small difference in mitochondrial activity between the two cell lines, and the HP cells showed increased membrane damage. In the HP cells, increased accumulation of Ca was concomitant with lower accumulation of K. We conclude that, while increased putrescine accumulation may have a protective role against ROS in plants, enhanced turnover of putrescine actually can make them vulnerable to increased oxidative damage. PMID- 19136267 TI - Storage oil hydrolysis during early seedling growth. AB - Storage oil breakdown plays an important role in the life cycle of many plants by providing the carbon skeletons that support seedling growth immediately following germination. This metabolic process is initiated by lipases (EC: 3.1.1.3), which catalyze the hydrolysis of triacylglycerols (TAGs) to release free fatty acids and glycerol. A number of lipases have been purified to near homogeneity from seed tissues and analysed for their in vitro activities. Furthermore, several genes encoding lipases have been cloned and characterised from plants. However, only recently has data been presented to establish the molecular identity of a lipase that has been shown to be required for TAG breakdown in seeds. In this review we briefly outline the processes of TAG synthesis and breakdown. We then discuss some of the biochemical literature on seed lipases and describe the cloning and characterisation of a lipase called SUGAR-DEPENDENT1, which is required for TAG breakdown in Arabidopsis thaliana seeds. PMID- 19136268 TI - Leucoanthocyanidin reductase and anthocyanidin reductase gene expression and activity in flowers, young berries and skins of Vitis vinifera L. cv. Cabernet Sauvignon during development. AB - Proanthocyanidins, or condensed tannins, are crucial polyphenolic compounds for grape and wine quality. Recently, significant advances were achieved in understanding the biosynthesis of their main subunits: (+)-catechin and (-) epicatechin, produced by catalysis of leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), respectively. Expression studies had been published but no data were available on enzyme activity. In our work, we devised assays to measure LAR and ANR activity and determine their development throughout the growth of flowers, young berries, and skins of Vitis vinifera L. cv. Cabernet Sauvignon. We also investigated the accumulation of compounds in these tissues and focused on the expression of both the structural genes and the transcription factors involved in regulating them: VvMYB5a and VvMYBPA1. Biosynthetic genes were expressed early and LAR and ANR were already active during flowering and at the beginning of berry growth, as well as during colour-change in skins. The profiles we determined correlated with total tannin, catechin, and epicatechin concentrations. The involvement of VvMYB5a and VvMYBPA1 was confirmed and specific expression patterns were also established for VvLAR transcripts. PMID- 19136269 TI - Characterisation of a new reporter system allowing high throughput in planta screening for recombination events before and after controlled DNA double strand break induction. AB - DNA double strand breaks (DSBs) are created either by DNA damaging reagents or in a programmed manner, for example during meiosis. Homologous recombination (HR) can be used to repair DSBs, a process vital both for cell survival and for genetic rearrangement during meiosis. In order to easily quantify this mechanism, a new HR reporter gene that is suitable for the detection of rare recombination events in high-throughput screens was developed in Arabidopsis thaliana. This reporter, pPNP, is composed of two mutated Pat genes and has also one restriction site for the meganuclease I-SceI. A functional Pat gene can be reconstituted by an HR event giving plants which are resistant to the herbicide glufosinate. The basal frequency of intra-chromosomal recombination is very low (10(-5)) and can be strongly increased by the expression of I-SceI which creates a DSB. Expression of I-SceI under the control of the 35S CaMV promoter dramatically increases HR frequency (10,000 fold); however the measured recombinant events are in majority somatic. In contrast only germinal recombination events were measured when the meganuclease was expressed from a floral-specific promoter. Finally, the reporter was used to test a dexamethasone inducible I-SceI which could produce up to 200x more HR events after induction. This novel inducible I-SceI should be useful in fundamental studies of the mechanism of repair of DSBs and for biotechnological applications. PMID- 19136270 TI - Regulation of de novo fatty acid synthesis in maturing oilseeds of Arabidopsis. AB - As a Brassicaceae, Arabidopsis thaliana constitutes an excellent model system to investigate oil biosynthesis in seeds. Extensive tools for the genetic and molecular dissection of this model species are now available. Together with analytical procedures adapted to its tiny seeds, these tools have allowed major advances in isolating and characterising the factors that participate in the metabolic and developmental control of seed filling. Once the biochemical pathways producing storage lipids, namely triacylglycerols, were elucidated, the question of the regulation of this metabolic network has arisen. The coordinated up regulation of genes encoding enzymes of the fatty acid biosynthetic pathway observed at the onset of seed maturation suggests that the pathway may be subjected to a system of global transcriptional regulation. This has been further established by the study of master regulators of the maturation program like LEAFY COTYLEDON2 and the characterisation of the WRINKLED1 transcription factor. These factors have been shown to participate in a regulatory cascade controlling the induction of the genes involved in fatty acid biosynthesis at the onset of the maturation phase. Although much remains to be elucidated, the framework of the regulatory system controlling fatty acid biosynthesis in Arabidopsis seeds is coming into focus. PMID- 19136272 TI - Video-assisted thoracic surgery lobectomy: can we afford it? AB - INTRODUCTION: Although video-assisted thoracoscopic (VATS) lobectomy offers advantages with regard to pain, respiratory function and mobility, concern is often expressed concerning the cost of stapling devices and increased operative time. We have therefore compared the costs of VATS and open lobectomy in order to assess the overall economic sustainability of a VATS programme. MATERIALS AND METHODS: Between January 2004 and December 2006 346 patients underwent pulmonary lobectomy mainly for stage I or II lung cancer (93 VATS lobectomy, 253 thoracotomy). In the VATS group 47% of patients had an upper lobectomy versus 52% in the open group (p=ns). Direct medical costs (disposables, theatre time, high dependency unit stay, hospital stay) were determined and stratified by lobectomy type. RESULTS: Mean theatre cost for a VATS lobectomy was 2533+/-230 euro versus 1280+/-54 euro for a thoracotomy lobectomy (p=0.00001). Mean high dependency unit (HDU) cost was 1713+/-236 euro and 2571+/-80 euro for a VATS and a thoracotomy lobectomy, respectively (p=0.00001). Mean cost of hospital stay for a VATS lobectomy was 3776+/-281 euro versus 4325+/-154 euro for an open one (p=0.00001). The overall cost for a VATS lobectomy was 8023+/-565 euro which was less than an open lobectomy 8178+/-167 euro (p=0.0002). VATS bilobectomy was slightly more expensive than an open one: 8702+/-350 euro versus 8655+/-466 euro but this difference (47 euro) was not significant (p=ns). CONCLUSION: VATS lobectomy is less expensive than conventional lobectomy. Increased theatre costs (disposables and time) are counteracted by shorter hospital stay. Importantly, the reduced HDU and ward bed stays free resources for other patients. PMID- 19136271 TI - Correlations between selected tumor markers and fluorodeoxyglucose maximal standardized uptake values in esophageal cancer. AB - OBJECTIVE: Esophageal cancer tumor biology is best assessed clinically by 2 [18F]fluoro-2-deoxy-d-glucose (FDG)-PET. Both FDG-PET maximal positron emission tomography (PET) standardized uptake values (SUVmax) and selected tumor markers have been shown to correlate with stage, nodal disease, and survival in esophageal cancer. Interestingly, there is limited data examining the relationship between FDG-PET SUVmax and expression of these tumor markers in esophageal cancer. The purpose of this study was to determine the correlation of tumor markers with FDG-PET SUVmax in esophageal cancer. METHODS: FDG-PET SUVmax was calculated in 67 patients with esophageal cancer of which 59 (88%) had adenocarcinoma. Neoadjuvant radiotherapy and/or chemotherapy were administered to 42% (28/67) of patients. Esophageal tumor tissue and surrounding normal tissue was obtained and tissue microarrays were created. Immunohistochemical analysis was performed for five known esophageal cancer tumor markers (GLUT-1, p53, cyclin D1, epidermal growth factor receptor (EGFR), and vascular endothelial growth factor (VEGF)). Assessment of each tumor marker was made by two independent, blinded pathologists using common grading criteria of intensity and percentage of cells stained. A p value <0.05 was considered significant. RESULTS: There were 55 men (82%) and 12 women (18%) with a median age of 63 years (range 40-83). Pathologic staging included stage I (n=29, 43%), stage II (n=19, 28%), stage III disease (n=18, 27%), and stage IV disease (n=1, 2%). PET SUVmax correlated with T stage (p=0.001). In patients undergoing surgery without induction therapy, increasing SUVmax values correlated with increased expression of GLUT-1 transporter (p=0.01). There was no correlation between SUVmax and EGFR, cyclin D1, VEGF, or p53 expression in primary tumor. CONCLUSIONS: FDG-PET SUVmax correlates with an increased expression of GLUT-1 transporter in esophageal cancer specimens not subjected to induction therapy. No significant difference in tumor marker expression was noted between patients undergoing induction therapy or surgery alone except p53 expression decreased in primary tumors following induction therapy. Failure of SUVmax values to correlate with known prognostic esophageal cancer tumor markers suggests that FDG-PET may have limited clinical utility in assessing response to therapies targeting these markers. PMID- 19136273 TI - Lung metastases from colorectal cancer: surgical resection and prognostic factors. AB - OBJECTIVE: To analyse our experience with excision of lung metastases from colorectal carcinoma (CRC), and to evaluate clinically relevant prognostic factors, identifying the cluster of patients who would benefit from this procedure. METHODS: Sixty-one patients, 42 men (69%), with primary CRC who underwent 94 curative resections of pulmonary metastases were retrospectively reviewed. Age was 30-80 years (mean 61.2+/-15). Population was analysed for age, sex, disease-free interval (DFI), prethoracotomy carcinoembryonic antigen (CEA) level, location and histology of primary tumour, number of lung lesions (and size of largest resected metastasis), type of lung resection, nodal involvement (hilar/mediastinal), use of adjuvant treatment, morbid-mortality and immediate and follow-up survival. RESULTS: Mean DFI was 29+/-22 months (range 5-132 months). There was no hospital mortality and significant morbidity occurred in five patients (8.2%). Mean follow-up was 39+/-4 months (range 4-173 months). Mean overall survival and disease-free survival were 67+/-16 months and 52+/-6 months, respectively. Three-, 5- and 10-year survival rates from date of primary colorectal resection were 83%, 71% and 43%, respectively. Three-, 5- and 10-year survival rates from date of lung resection were 61%, 48% and 11%, respectively. Five-year survival was 57% in patients with normal prethoracotomy CEA levels and 18% for those with high levels (>5 ng/ml) (p=0.039). CONCLUSIONS: Pulmonary metastasectomy has potential survival benefit for patients with metastatic colorectal carcinoma. Low morbidity and mortality rates, contrasting with lack of any other effective therapy, justify aggressive surgical management. Single deposits, DFI >36 months and normal prethoracotomy serum CEA were significant independent prognostic factors. PMID- 19136274 TI - Short-term systolic and diastolic ventricular performance after surgical ventricular restoration for dilated ischemic cardiomyopathy. AB - OBJECTIVE: Based on the adverse relationship between left ventricular (LV) remodeling and clinical outcome in ischemic cardiomyopathy, surgical ventricular restoration (SVR) is proposed as a valuable adjunct procedure. This study reports on the short-term clinical and hemodynamical performance of SVR. METHODS: Using end-systolic LV volume as indication for SVR, 78 patients with ischemic cardiomyopathy are divided in two groups: group 1 comprised 55 patients treated by coronary revascularization and mitral annuloplasty, group 2 comprised 23 patients undergoing additional SVR. Hemodynamic investigation included echocardiographic assessment of systolic and diastolic function. Clinical follow up focused on survival and functional status with exercise performance. RESULTS: Both surgical approaches resulted in improvement of NYHA class (2.9-1.6 in group 1; 3.3-1.5 in group 2, p<0.001), achieving similar exercise performance (peak VO2 13.7 vs 15.4 ml/kgmin in groups 1 and 2, p=0.25) and plasma BNP values (group 1: 1350 pg/ml and group 2: 767 pg/ml, p=0.23). SVR provided additional benefit as patients basically had a worse NYHA class (2.9 in group 1 vs 3.3 in group 2, p=0.03). Within mean follow-up of 20 months, survival rate was 84% in group 1 and 74% in group 2 (p=0.11), including operative mortality of 7% and 13% (p=0.42). Through effective volume reduction (LVEDVI 41%; LVESVI 49%) systolic function improved immediately after SVR (LVEF 27-39% in group 2, p<0.05). Worsening of diastolic function was specifically observed after SVR within the first year (E/A ratio 1.38-1.74 cm/s, p=0.02). Recurrent mitral regurgitation (p=0.004) and secondary remodeling (p=0.01) were major determinants of decreasing LV compliance. Clinical outcome in terms of cardiac events and survival was compromised by restrictive diastolic function (p=0.02) and increased LV volumes (p=0.04). CONCLUSION: SVR in addition to coronary revascularization and restrictive mitral annuloplasty results in significant clinical improvement in selected patients with advanced ischemic heart disease and severely dilated ventricles. SVR entails immediate improvement of systolic function, which remains sustained during short-term follow-up. Serial assessment of diastolic function is mandatory as LV compliance seems more sensitive to early changes induced by recurrence of mitral regurgitation and secondary ventricular dilation. Moreover, worsening of diastolic dysfunction should be timely recognized because of its adverse clinical impact. PMID- 19136275 TI - Plasmapheresis before thymectomy in myasthenia gravis: routine versus selective protocols. AB - OBJECTIVE: The value of thymectomy in management of myasthenia gravis (MG) is well recognized. Plasmapheresis (PMP) before thymectomy appears to improve the postoperative outcome. As PMP has its own complications, selective use of PMP preoperatively for patients at risk of post-thymectomy complications would improve the postoperative outcome, and decreases the PMP-related complications. The aim of this study is to evaluate the effectiveness and safety of routine versus selective use of PMP before thymectomy. MATERIAL AND METHODS: We performed a retrospective analysis of two different protocols in two institutions comparing the routine (group I) versus selective use (group II) of prethymectomy PMP. The computerized database and the medical records of 164 patients diagnosed with MG who underwent thymectomy between 1998 and 2007, were reviewed. RESULTS: In group I, 74 patients were treated with PMP before thymectomy. In group II (90 patients), 35 patients were identified as high-risk and were treated with PMP before thymectomy, and in 55 patients, thymectomy was performed without preoperative PMP. There was significant difference in the recorded PMP-related complications between group I and group II; 25.7% (19 patients) versus 8.9% (8 patients), respectively. There was no difference between the two groups as regards the duration of postoperative mechanical ventilation, intensive care unit (ICU) stay and hospital stay. CONCLUSION: This study demonstrated that selective use of PMP before thymectomy may reduce the incidence of PMP-related complications without affecting the overall outcome. PMID- 19136276 TI - Diamino acid derivatives of PpIX as potential photosensitizers for photodynamic therapy of squamous cell carcinoma and prostate cancer: in vitro studies. AB - Photodynamic therapy (PDT) is an alternative treatment modality involving light activated drugs, called photosensitizers (PSs), to treat cancer and non-cancerous conditions. The search for new compounds which might become effective PSs is the major direction for PDT development. In the present work we have studied the dark toxicity, intracellular localization and photodynamic properties of four potential, water soluble, second generation PSs--PP(Arg)(2), PP(Ser)(2)Arg(2), PP(Ala)(2)Arg(2), PP(Phe)(2)Arg(2), all diamino acid derivatives of protoporphyrin IX. Human prostate cancer (DU-145) and squamous carcinoma (A431) cells were used as experimental model. Among investigated compounds PP(Ser)(2)Arg(2) exhibited the lowest dark toxicity and the highest PDT effectiveness towards both cell lines. Fluorescence microscopy revealed the time dependent changes in intracellular localization of the PS which were related to the phototoxicity. The results show that PP(Ser)(2)Arg(2) may be a potential PS for PDT. PMID- 19136277 TI - Biomodulation with low-level laser radiation induces changes in endothelial cell actin filaments and cytoskeletal organization. AB - The cytoskeleton is a central and vital structure of eukaryotic cells. It consists of a dynamic network of partially interconnected polymers. This extended network controls the mechanical properties of animal cells, serves as intracellular transport "pathways", and plays a prominent role in cell motility, proliferation, and adhesion. In order to evaluate the action of laser irradiation on the cytoskeleton and proliferation of endothelial cells, rabbit aortic endothelial cells (RAEC) were irradiated with 685-nm low-level laser (20 mW output power). Fluorescent dye rhodamine-phalloidin staining was used to visualize the effect of laser irradiation on actin filaments. Irradiation with 8 J/cm(2) was performed four times at 12-h intervals for 24 min. Cells cultured under low fetal bovine serum condition (5% FBS) for 7 days presented actin staining predominantly in the cortical membrane region and a few actin filament stress fibers. However, the formation of stress fibers similar to those of control cells increased significantly in irradiated cells. It was concluded that laser irradiation induces changes in the cytoskeleton of endothelial cells through the reorganization of actin filaments and neo-formation of stress fibers, allowing evident cellular proliferation. PMID- 19136278 TI - Transforming growth factor beta 1 (TGFbeta1) gene polymorphisms and Chagas disease susceptibility in Peruvian and Colombian patients. AB - Susceptibility to Chagas disease infections and its clinical manifestations may be influenced by host genetic factors. Among cytokines, the multifunctional transforming growth factor beta 1 (TGFbeta1), plays a major role in the establishment and pathogenesis of Trypanosoma cruzi infection, the etiological agent of Chagas disease. Several single-nucleotide polymorphisms (SNP) in the TGFbeta1 gene that may affect cytokine production have been described. We investigated, by PCR methods, five SNP in the TGFbeta1 gene of known or suggested functional significance (-988 C/A; -800 G/A; -509 C/T; 10 T/C and 263 C/T) in 347 seropositive (asymptomatic, n=175; cardiomyopathic, n=172) and 279 seronegative unrelated individuals from a Peruvian and a Colombian population where T. cruzi is highly endemic. We found a significant difference in the distribution of the TGFbeta1 10T and 10C alleles between patients and healthy controls in both cohorts, analyzed independently and combined. The frequency of the high TGFbeta1 producer genotype 10 C/C was increased in the patients groups of both populations. Our data suggests that TGFbeta1 genetic polymorphisms at codon 10 may be involved in a differential susceptibility to T. cruzi infection in these South American samples. PMID- 19136279 TI - Functional independence of explicit and implicit motor adjustments. AB - Adaptation to novel visuomotor transformations for example when navigating a cursor on a computer monitor by using a computer mouse, can be explicit or implicit. Explicit adjustments are made when people are informed about the occurrence and the type of a novel visuomotor transformation and intentionally modify their movements. Implicit adjustments, in contrast, are made without reportable knowledge of a novel visuomotor transformation and without a change intention. The relation of implicit adjustments to explicit adjustments needs further clarification. Here we show that these two types of adjustment occur at the same time and remain functionally independent. The size of total adjustment turned out to be the sum of explicit and implicit adjustments measured in isolation, even when both processes produce opposite outcomes. In perspective our results demonstrate that automatic, implicit processes of motor control are not superseded by intentional, explicit ones, but only superposed. PMID- 19136280 TI - Temperament and intuition: a commentary on Feltz and Cokely. AB - In this paper, we examine Adam Feltz and Edward Cokely's recent claim that "the personality trait extraversion predicts people's intuitions about the relationship of determinism to free will and moral responsibility" (INSERT REFERENCE). We will first present some criticisms of their work before briefly examining the results of a recent study of our own. We argue that while Feltz and Cokely have their finger on the pulse of an interesting and important issue, they have not established a robust and stable connection between extraversion and compatibilist-friendly intuitions. PMID- 19136281 TI - Mode of vagus nerve stimulation differentially affects sleep related breathing in patients with epilepsy. AB - PURPOSE: We describe the influence of vagus nerve stimulation (VNS) with standard mode and rapid cycling mode on sleep related breathing in two patients with epilepsy. METHODS: Two VNS treated patients underwent digital video polysomnography for three nights (night 1: rapid cycling mode; night 2: standard mode; night 3: off mode). RESULTS: In patient 1, on off mode, apnea-hypopnea index (AHI) was 11.1/h, respiratory effort-related arousal index (RERAI) 0.9/h, flow limitation index (FLI) 0.9/h and oxygen desaturation index (ODI) 10.2/h. On standard mode, AHI was 5.5/h, RERAI 1.7/h, FLI 4.1/h and ODI 5.5/h. On rapid cycling mode, AHI was 10.4/h, RERAI 7.9/h, FLI 17.3/h and ODI 10.3/h. In patient 2, on off mode, AHI was 1.6/h, RERAI 0.8/h, FLI 2.2/h and ODI 0/h. On standard mode, AHI was 2.9/h, RERAI 2.4/h, FLI 2.6/h and ODI 2.9/h. On rapid cycling mode, AHI was 0.7/h, RERAI increased to 15.4/h, FLI to 52.0/h and ODI was 0.7/h. CONCLUSIONS: The number of RERAs as well as the number of flow limitations were higher with the rapid cycling mode compared to standard mode and stimulation off and might be related to the higher impulse frequency. PMID- 19136282 TI - Posterior cortex epilepsy: diagnostic considerations and surgical outcome. AB - PURPOSE: To assess the role of various pre-surgical evaluations in posterior cortex epilepsy (PCE) and its surgical outcome. METHODS: A retrospective analysis of clinical and laboratory data from 43 PCE patients referred for surgery was performed. The diagnosis was established by standard pre-surgical evaluation modalities including semiology, MRI, interictal and ictal scalp video-EEG as well as additional intracranial EEG monitoring in selected cases. RESULTS: The 43 patients included 11 parietal lobe epilepsies, 13 occipital lobe epilepsies, and 19 patients with seizures originating from parieto-occipito-posterior temporal cortex. Thirty-three (76.7%) patients experienced at least one type of aura. Seventeen patients showed complex focal seizures, which were followed by secondarily generalized tonic-clonic seizures in seven of them; eighteen patients predominantly showed simple motor seizures (clonic seizures in 6, tonic seizures in 7, and versive seizures in 5). Long-term visual field deficits were observed in 8 patients. Other transient neurological deficits occurred in 7 patients. All patients received the follow-up study lasting 1-5 years, and achieved Engel's I in 26 cases, II in 5, III in 5, and IV-V in 7. Malformation of cortical development was diagnosed in 41.9% of our surgical population. No significant relationship was found between the diagnostic accuracy of any pre-surgical evaluating modality and surgical outcome in this series. CONCLUSIONS: Surgical treatment is effective for PCE. Accurate localization of epileptogenic zone and eloquent cortex are two key factors for favorable outcome. None of the diagnostic modalities shows obvious predictive value for favorable surgical outcome. PMID- 19136283 TI - Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation. AB - OBJECTIVE: The effects of inflammation on bone development from mesenchymal stem cells (MSC) are unclear due to the difficulty in isolating MSC. The aim of this study was to develop a MSC isolation method and to determine the in vitro effects of interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) on their osteogenic differentiation. METHODS: Murine MSC were isolated from the limbs of C57/Bl6 mice through collagenase digestion of bone and enriched as the Stem cell antigen (Sca-1)(+) CD31(-) CD45(-) population, using lineage immunodepletion, followed by fluorescence-activated cell sorting (FACS). They were differentiated along the osteoblast linage in the presence or absence of IL 1beta and TNFalpha. Mineralization was measured as was the expression of a number of osteogenic genes by quantitative polymerase chain reaction (PCR). RESULTS: We show that osteogenic differentiation from the MSC population is suppressed by IL 1beta and TNFalpha. In addition to suppression of bone mineralization, both cytokines inhibited the differentiation-associated increases in alkaline phosphatase (ALP) activity and the gene expression for ALP, alpha1(I) procollagen, runt-related transcription factor 2 (Runx2) and osterix. However, only TNFalpha inhibited osteonectin and osteopontin mRNA expression and only IL 1beta reduced cell proliferation. CONCLUSIONS: The convenient isolation technique enables the easy generation of sufficient MSC to permit the molecular analysis of their differentiation. We were thus able to show that the proinflammatory cytokines, IL-1beta and TNFalpha, can compromise bone development from this primary MSC population, although with some significant differences. The potential involvement of specific inflammatory mediators needs to be taken into account if optimal bone repair and presumably that of other tissues are to be achieved with MSC. PMID- 19136284 TI - The potential role of MRI in veterinary clinical cardiology. AB - Over the last decade, magnetic resonance imaging (MRI) has become established as a useful referral diagnostic method in veterinary medicine that is widely used in small animal brain and spinal diseases, aural, nasal and orbital disorders, planning soft tissue surgery, oncology and small animal and equine orthopaedics. The use of MRI in these disciplines has grown due to its unparalleled capability to image soft tissue structures. This has been exploited in human cardiology where, despite the inherent difficulties in imaging a moving, contractile structure, cardiac MRI (CMRI) has become the optimal technique for the morphological assessment and quantification of ventricular function. Both CMRI hardware and software systems have developed rapidly in the last 10 years but although several preliminary veterinary CMRI studies have been reported, the technique's growth has been limited and is currently used primarily in clinical research. A review of published studies is presented with a description of CMRI technology and the potential of CMRI is discussed along with some of the reasons for its limited usage. PMID- 19136285 TI - Neural tube defect in a 4000-year-old Egyptian infant mummy: a case of meningocele from the museum of anthropology and ethnography of Turin (Italy). AB - This paper reports a paleopathological study of a severe neural tube defect in an ancient mummy, more specifically, a meningocele in an Egyptian infant from the XI dynasty (2100-1955B.C.). This is one of the most ancient cases of meningocele in mummified human remains described in paleopathological literature. Prehistoric and early historic examples of severe congenital defects of the vertebral column and neural tube are rare, because of the precarious preservation conditions of ancient human remains. Further, since the majority are only the skeletal remains, paleopathological and paleoepidemiological analysis based on the observation of bones is even more difficult. Hence, it is not easy to investigate this disease in the past in all its complexities and true diffusion. The case presented here is peculiar, since it concerns a mummy with almost all soft tissues preserved, thus allowing us to describe the defect in an infant. Only targeted, minimally invasive examinations were performed. An anthropological investigation with helical CT scan and histological analysis was used to diagnose the defect and identify post-mortem transformation processes. The analyses confirmed the diagnosis of meningocele in an approximately six-month-old infant. PMID- 19136286 TI - Stress and thermoregulation: different sympathetic responses and different effects on experimental pain. AB - Stress and thermoregulation both activate the sympathetic nervous system (SNS) but might differently affect pain. Studies investigating possible interactions in patients are problematic because of the high prevalence of SNS disturbances in patients. We therefore analyzed the influence of these different sympathetic challenges on experimentally-induced pain in healthy subjects. SNS was activated in two different ways: by mental stress (Stroop task, mental arithmetic task), and by thermoregulatory stimulation using a water-perfused thermal suit (7 degrees C, 32 degrees C, or 50 degrees C). Attentional effects of the mental stress tasks were controlled by using easy control tasks. Both, stress and thermoregulatory stimuli, robustly activated SNS parameters. However, the patterns of activation were different. While stress co-activated heart rate, blood pressure, peripheral vasoconstriction and sweating, thermal stimulation either increased blood pressure (cold) or heart rate and sweating (warm). Only stress was able to induce a significant reduction of pain. The control tasks neither activated the SNS nor altered pain perception. Our results suggest that (1) different patterns of sympathetic activation can be recorded after stress and thermoregulatory challenges and (2) that only stress is able to interfere with sensation of experimental pain. Whether SNS activation is causally responsible for analgesia needs to be further investigated. PMID- 19136287 TI - Toxicity of peroxisomal C27-bile acid intermediates. AB - Peroxisomes play an important role in bile acid biosynthesis because the last steps of the synthesis pathway are performed by the beta-oxidation system located inside peroxisomes. As a consequence, C(27)-bile acid intermediates accumulate in several peroxisomal disorders. It has been suggested that C(27)-bile acids are especially toxic and contribute to the liver disease associated with peroxisomal disorders. For this reason, we investigated the toxicity of C(27)-bile acids and the underlying mechanisms. We studied the effects of conjugated and unconjugated C(27)-bile acids on cell viability, mitochondrial respiratory chain function and production of oxygen radicals in the rat hepatoma cell line McA-RH7777. Cell viability decreased progressively after incubation with increasing concentrations of different bile acids with dihydroxycholestanoic acid (DHCA) being clearly the most cytotoxic bile acid. In addition, the different bile acids caused a dose dependent decrease in ATP synthesis by isolated mitochondria oxidizing malate and glutamate. Finally, there was a dose-dependent stimulation of ROS generation in the presence of C(27)-bile acids. In conclusion, our studies showed that C(27) bile acids are more cytotoxic than mature C(24)-bile acids. In addition, C(27) bile acids are potent inhibitors of oxidative phosphorylation and enhance mitochondrial ROS production by inhibiting the respiratory chain. PMID- 19136288 TI - Infection outcomes in splenectomized patients with hemoglobinopathies in Australia. AB - OBJECTIVE: To assess the infection risk of splenectomized persons with hemoglobinopathies in Australia. METHODS: This was an Australia cohort study of infections and vaccinations in 63 patients with hemoglobinopathies followed longitudinally from 1967 to 2006, and an investigation into the risk factors for poor outcome. RESULTS: There were 28 cases of bacterial infection that necessitated hospitalization in the cohort, resulting in an incidence of 1.4 bacterial infections per 100 patient-years. There was one death (1.6%) as a direct result of bacterial infection. Hepatitis C was diagnosed in 28 patients (44%). The spectrum of infection included pneumonia (6/28), cellulitis (6/28), bacteremia (4/28), and skin abscess (3/28). Notably, Klebsiella species organisms were isolated in 9/28 cultures. CONCLUSIONS: Infectious complications in this group of patients cause serious morbidity and mortality. This cohort may be a target for novel preventive strategies such as more immunogenic vaccines, patient registries, and/or education programs. PMID- 19136289 TI - Understanding transmitted HIV resistance through the experience in the USA. AB - Transmitted drug resistance is an emerging phenomenon with important clinical and public health implications. It has been reported in 3.4% to 26% of HIV-infected persons in the USA. Most cases affect non-nucleoside reverse transcriptase inhibitors or nucleos(t)ide reverse transcriptase inhibitors. Transmitted protease inhibitor or multi-class resistance is uncommon, occurring in <5% of cases. The genital tract may function as a reservoir of transmissible drug resistant variants or a site for low-level viral replication at a time plasma HIV is suppressed. Transmitted drug-resistant HIV variants, including those that exist in very low titers (minority populations), are associated with suboptimal virologic response to initial antiretroviral therapy. Baseline resistance testing, preferably genotype, appears to be cost-effective and is recommended for all treatment-naive patients in the USA, although prospective trials have not been performed. It appears transmitted drug resistance is still relatively low in developing countries, but there is a dearth of information. PMID- 19136290 TI - Verification of Berlin algorithm for diagnosing undifferentiated spondyloarthropathy patients in Chinese population. AB - OBJECTIVE: To evaluate the diagnosing value of the Berlin algorithm, comparing to that of the established ESSG and Amor criteria in Chinese patients with undifferentiated spondyloarthropathy. METHODS: A total of 92 clinically diagnosed undifferentiated spondyloarthropathy patients with axial involvement were compared to 123 patients with other kinds of rheumatic diseases by using the parameters listed in the Berlin algorithm, ESSG criteria, and Amor criteria. RESULTS: In the 92 undifferentiated spondyloarthropathy patients with axial involvement, the prevalence rate of HLA-B27 was 71.76% (61/85). Elevated ESR and/or CRP was found in 40.96% (34/83) SpA patients and abnormal MRI manifestation of sacroiliac joint was found in 91% (39/43) SpA patients. The specificity of HLA-B27 was 78% and similar with the Berlin study. The sensitivity/specificity of ESSG, Amor criteria and Berlin algorithm on diagnosing USpA was 72.83%/92.68%, 64.13%/93.50% and 67.39%/95.93%, respectively. The coincidence between the three diagnosing criteria and the rheumatologist's opinion was moderate. CONCLUSION: Our study showed the new Berlin algorithm has important value of diagnosing undifferentiated spondyloarthropathy in China, which has the similar diagnosing capacity comparing to the traditional criteria ESSG and Amor criteria. PMID- 19136291 TI - Cadmium and lead ions can be removed simultaneously from a binary aqueous solution by the sono-sorption method. AB - This paper presents the effect of ultrasound on the simultaneous removal of Cd(2+) and Pb(2+) ions from a binary aqueous solution in the presence of natural sorbent (sono-sorption method). The obtained results compare with the results of control method which was achieved in the absence of ultrasound. The experiments under sonication were carried out by 20 kHz apparatus. The effect of different parameters such as the amount of sorbent, contact time, temperature, and specially the competitive sorption of ions were considered for both methods. It is found that the sorption efficiency of Cd(2+) was decreased in the presence of Pb(2+). But, the presence of Cd(2+) had a negligible effect on the removal of Pb(2+). The results show that in the binary solution, the removal efficiency was higher in the presence of ultrasound than the control method. Both ions in binary solution can be removed to near completion with ultrasound. Two different multi component isotherm equations based on the Langmuir isotherm were applied on the studied system to predict the sorption behavior of the components. PMID- 19136292 TI - A finite element analysis of the vibrational behaviour of the intra-operatively manufactured prosthesis-femur system. AB - In total hip replacement (THR) a good initial stability of the prosthetic stem in the femur, which corresponds to a good overall initial contact, will help assure a good long-term result. During the insertion the implant stability increases and, as a consequence, the resonance frequencies increase, allowing the assessment of the implant fixation by vibration analysis. The influence of changing contact conditions on the resonance frequencies was however not yet quantitatively understood and therefore a finite element analysis (FEA) was set up. Modal analyses on the hip stem-femur system were performed in various contact situations. By modelling the contact changes by means of the contact tolerance options in the finite element software, contact could be varied over the entire hip stem surface or only in specific zones (proximal, central, distal) while keeping other system parameters constant. The results are in agreement with previous observations: contact increase causes positive resonance frequency shifts and the dynamic behaviour is most influenced by contact changes in the proximal zone. Although the finite element analysis did not establish a monotonous relationship between the vibrational mode number and the magnitude of the resonance frequency shift, in general the higher modes are more sensitive to the contact change. PMID- 19136293 TI - Synthetic organic pigments of the 20th and 21st century relevant to artist's paints: Raman spectra reference collection. AB - Some 170 organic pigments relevant to artist's paints have been collected from historic collections and modern manufacturers. The number includes multiples of the same pigment from different sources and comprises 118 different color indices (C.I.). All of them have been analysed with FTIR spectroscopy and 125 pigments (93 different C.I. No.s) of particular relevance to artist's paints have been characterised with Raman spectroscopy so far. The pigment collection encompasses the following pigment classes and subgroups: monoazo pigments represented by acetoacetic arylide yellow (hansa yellow), beta-naphthol, BON, naphthol AS and benzimidazolone; disazo pigments with disazo condensation, diarylide, bisacetoacetarylide, pyrazolone; azo-azomethin metal complex pigments; non-azo, polycyclic pigments such as phthalocyanines, diketopyrrolo-pyrroles (DPP), perylenes and perinones, quinacridones, isoindolinones, polycarbocyclic anthraquinones and dioxanines. The selection of references was based on availability (historic collections) and current use in 16 acrylic, alkyd and oil based artist's paints, and it covers pigment colors PY yellow (27 C.I. No.s), PR red (38), PO orange (9), PB blue (8), PV violet (6), PG green (3) and PBr brown (2). Besides peak tables and spectra patterns, flow charts based on color, pigment class, group and individual color index are presented to help identification of unknowns and mixed paint samples. While Raman could isolate all different C.I. numbers, multiple references of the same C.I. from different sources could not be distinguished. PMID- 19136294 TI - Adaptation and vision change the relationship between muscle activity of the lower limbs and body movement during human balance perturbations. AB - OBJECTIVE: Investigate the relationship between changes in lower limb EMG root mean square (RMS) activity and changes in body movement during perturbed standing. Specifically, linear movement variance, torque variance and body posture were correlated against tibialis anterior and gastrocnemius RMS EMG activity during perturbed standing by vibration of the calf muscles. METHODS: Eighteen healthy participants (mean age 29.1 years) stood quietly for 30s before vibration pulses were randomly applied to the calf muscles over a period of 200 s with eyes open or closed. Movement variance, torque variance and RMS EMG activity were separated into five periods, thereby allowing us to explore any time-varying changes of the relationships. RESULTS: Changes of tibialis anterior muscles EMG activity were positively correlated with changes in linear movement variance and torque variance throughout most of the trials, and negatively correlated with some mean angular position changes during the last 2 min of the trials. Moreover, the initial changes in Gastrocnemius EMG activity were associated with initial changes of mean angular position. Additionally, both tibialis anterior and gastrocnemius muscle activities were more involved in the initial control of stability with eyes closed than with eyes open. CONCLUSIONS: Visual information and adaptation change the association between muscle activity and movement when standing is perturbed by calf muscle vibration. SIGNIFICANCE: Access to visual information changes the standing strategy to calf muscle vibrations. Training evoking adaptation could benefit those susceptible to falls by optimising the association between muscle activities and stabilising body movement. PMID- 19136295 TI - General physical activity levels influence positive and negative priming effects in young adults. AB - OBJECTIVE: To investigate the relationship between general physical activity level and the cognitive functions of executive control in young adults using behavioral measures and event-related brain potentials. METHODS: Forty young adults (mean age=21.1 yrs; 19 females) were differentiated on the basis of their regular physical activity level into two groups: active and sedentary. They performed a spatial priming task consisting of three conditions: control, positive, and negative priming. Spatial priming effects, which are related to executive control and occur automatically, were assessed as indicators of cognitive functioning. RESULTS: Negative priming effects on reaction time and P3 latency in the active group were larger than in the sedentary group. By contrast, positive priming effects were only observed in the sedentary group. CONCLUSIONS: The cognitive effects of regular physical activity could be observed using a relatively simple paradigm. The results indicate that regular physical activity has a beneficial effect on the cognitive processes on executive control in young adults. SIGNIFICANCE: The present study provides additional evidence of the beneficial effects of regular physical activity on cognitive functioning in young adults. PMID- 19136296 TI - The vestibular evoked myogenic potential: a test-retest reliability study. AB - OBJECTIVE: The purpose of this study was to evaluate the reliability of the vestibular evoked myogenic potential (VEMP) response in a clinical setting when only a feedback mechanism is available for monitoring background EMG. METHODS: Sixty-one healthy subjects participated in the investigation of the within session reliability, whereas fourteen of them were retested after one week for examination of the between-session reliability. VEMPs were recorded using 500 Hz tone bursts with the subjects in a sitting position and their heads turned away from the test ear to the contralateral shoulder, thereby pushing their chin against the inflatable cuff of a blood pressure manometer, serving as feedback method. RESULTS: The feedback method revealed latency and amplitude values comparable to other data in the literature where different test conditions were applied. Excellent reliability with ICC values ranging from 0.78 to 0.96 and CV(ME) values ranging from 4% to 36%, was achieved for P1 and N1 latency, threshold and interpeak amplitude. Good reliability with ICC values of 0.65 and 0.68 and CV(ME) values of 170% and 189%, was obtained for the asymmetry ratio. CONCLUSIONS: A unilateral muscle contraction controlled by a feedback mechanism resulted in reliable response parameters, comparable right to left and corresponding to literature data obtained in different test conditions. SIGNIFICANCE: The use of a blood pressure manometer as feedback mechanism combined with a meticulously controlled positioning of the head and contraction of the SCM muscle provides a reliable alternative in clinical settings, when the background muscle contraction cannot be measured or software related correction algorithms are not accessible. PMID- 19136297 TI - Methodology for intraoperatively eliciting motor evoked potentials in the vocal muscles by electrical stimulation of the corticobulbar tract. AB - OBJECTIVE: To establish a methodology for recording corticobulbar motor evoked potentials (CoMEPs) from vocal muscles after transcranial electrical stimulation (TES) and direct cortical stimulation (DCS). METHODS: Twenty-four patients were included in this study (22 for TES, 2 for DCS, 3 for TES plus DCS) that underwent different surgical procedures. We used two methods to elicit CoMEPs: (a) TES by stimulation over C3/Cz or C4/Cz and (b) DCS with a strip electrode placed over the primary motor area (M1) for laryngeal muscles. To record CoMEPs from vocal muscles we used two hook wire electrodes 76 microm of diameter passing through 27 gauge needle endotracheally placed in the vocal muscles after intubation. RESULTS: Recording of CoMEPs in the vocal muscles after TES was successfully performed in 22 patients. TES over the right or left hemisphere elicit responses bilaterally. The onset latencies for the right vocal muscle was 12.4+/-3.1 ms (ipsilateral stimulation) and 12.7+/-2.2 ms (contralateral stimulation) while for the left vocal muscle, onset latency was 12.9+/-2.3 ms (ipsilateral stimulation) and 14.1+/-3.4 ms (contralateral stimulation). In five patients DCS elicited CoMEPs in right and left vocal muscle with latency of 16.6+/-4.7 and 15.6+/-3.7 ms, respectively. CONCLUSION: The method to elicit and record CoMEPs in vocal muscles shows reliable results and adds one more tool in the armamentarium of intraoperative neurophysiology. SIGNIFICANCE: This method shows the ability to continuously monitor the functional integrity of corticobulbar pathways, vagal nucleus and laryngeal nerves. PMID- 19136298 TI - The involvement of cognitive processing in a perceptual-motor process examined with EEG time-frequency analysis. AB - OBJECTIVE: For motor activities, visual information is crucial for organizing a movement with respect to a given situation. The present study investigates how cognitive information processing is associated with this visuomotor process. METHODS: Brain dynamics in executing two perceptual-motor tasks were examined in terms of event-related synchronization (ERS) and event-related desynchronization (ERD) of EEG. Those tasks were (1) reaching toward and grasping a visual object with a pinch grip, and (2) matching the pinch grip size with respect to the perceived object size. RESULTS: According to the aperture size in the task execution, both the tasks were affected by the perceived object size inducing the Ebbinghaus illusion. The alpha-ERD patterns were associated with the movement execution and appeared to be identical in both the tasks, whilst the gamma-ERS appeared only for the grasping motion. CONCLUSIONS: These results suggest that cognitive processing was involved not only in the matching task but also in the grasping task. These ERD/ERS patterns are thought to reflect the similarity and difference in the perceptual-motor processes between the two tasks. SIGNIFICANCE: The analysis of ERD/ERS can provide insight on the qualitative feature in a visuomotor process associated with the involvement of cognitive processing. PMID- 19136299 TI - Sensory afferent inhibition within and between limbs in humans. AB - OBJECTIVE: To examine the distribution and inter-limb interaction of short latency afferent inhibition (SAI) in the arm and leg. METHODS: Motor evoked potentials (MEPs) in distal and proximal arm, shoulder and leg muscles induced with ranscranial magnetic stimulation (TMS) were conditioned by painless electrical stimuli applied to the index finger (D2) and great toe (T1) at interstimulus intervals (ISIs) of 15, 25-35, 80 ms (D2) and 35, 45, 55, 65 and 100 ms (T1) in 27 healthy human subjects. TMS was delivered over primary motor cortex (M1) arm and leg areas. Electrical stimulus intensities were varied between 1 and 3 times the sensory perception thresholds. We also tested effects of posterior cutaneous brachial nerve (PCBN) stimulation on MEPs in arm muscles at ISIs of 18 and 28 ms. RESULTS: D2 but not PCBN electrical conditioning reduced MEP amplitudes in upper limb muscles at ISIs of 25 and 35 ms. SAI was more pronounced in distal as compared to proximal arm muscles. Also, SAI following D2 stimulation increased with higher conditioning intensities. D2 stimulation did not change lower limb muscles MEPs. In contrast, T1 stimulation did not induce SAI in any muscles but caused MEP facilitation in a foot muscle at an ISI of 55 ms and in upper limb muscles at ISIs of 35 and 55 ms. Short interval intracortical inhibition (SICI) and intracortical facilitation (ICF) were not affected by electrical T1 conditioning. CONCLUSION: D2 stimulation causes segmental SAI in upper limb muscles with a distal to proximal attenuation without affecting leg muscles. In contrast, toe stimulation facilitates motor output both in foot and upper arm muscles. SIGNIFICANCE: Our data suggest that cutaneo-motor pathways in arms and legs are functionally organized in a different way with cutaneo-motor interactions induced by toe stimulation probably relayed at a thalamic level. Abnormal cutaneo-motor interactions following electrical toe stimulation may serve as an electrophysiological marker of thalamic dysfunction, e.g. in neurodegenerative diseases. PMID- 19136300 TI - Nocturnal sympathetic activity and hypertension: riding the wave of the peripheral pulse. PMID- 19136301 TI - Protection of free-living and captive possums against pulmonary challenge with Mycobacterium bovis following oral BCG vaccination. AB - In New Zealand, possums (Trichosurus vulpecula) are the main wildlife reservoir for bovine tuberculosis (Tb), which they transmit to livestock. This study investigated oral vaccination with lipid-formulated Mycobacterium bovis BCG and subsequent protection against virulent M. bovis challenge in wild-caught possums. Possums were trapped from the field and either hand-vaccinated and released back into the wild, or acclimatised to captive conditions prior to voluntary uptake of flavoured vaccine. Possums were subsequently exposed to pulmonary challenge with virulent M. bovis, administered either by instillation of a liquid suspension as an intra-tracheal challenge (field animals) or in micro-droplets as an aerosol (captive animals). Field studies indicated that the relative risk of death in wild possums due to Tb was 2.4 times greater in control compared with orally vaccinated possums, with the vaccine conferring protection to possums in both good and poor body condition. Laboratory studies indicated that oral vaccination conferred protection in cage-acclimatised possums, with >3log(10) reduction in lung bacterial burdens among vaccinated animals. This study provides evidence that lipid-formulated BCG oral vaccine can provide significant protection to possums in field as well as laboratory conditions, which may favour the use of this formulation as a delivery method for controlling wildlife Tb. PMID- 19136302 TI - Adjuvant methotrexate in CIDP. PMID- 19136303 TI - Randomised controlled trial of methotrexate for chronic inflammatory demyelinating polyradiculoneuropathy (RMC trial): a pilot, multicentre study. AB - BACKGROUND: Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) responds to treatment with corticosteroids, intravenous immunoglobulin, and plasma exchange. We aimed to test whether the standard immunosuppressive drug methotrexate was of use in treatment of CIDP. METHODS: In a pilot, multicentre, randomised, double-blind, controlled trial we compared oral methotrexate 7.5 mg weekly for 4 weeks, then 10 mg weekly for 4 weeks, and finally 15 mg weekly for 32 weeks (40 weeks' total treatment) with placebo in patients with CIDP requiring intravenous immunoglobulin or corticosteroids. After about 16 weeks, the dose of corticosteroids or intravenous immunoglobulin was decreased by 20% every 4 weeks if participants did not deteriorate. Primary outcome was a greater than 20% reduction in mean weekly dose in the last 4 weeks of the trial compared with the first 4 weeks. Secondary outcomes analysed separately at the mid-trial and final visits measured activity limitations and strength. Analyses were done by intention to treat. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN73774524. FINDINGS: 59 of the 60 enrolled participants completed the trial. 14 (52%) of 27 taking methotrexate and 14 (44%) of 32 taking placebo had a greater than 20% reduction in mean weekly dose of corticosteroids or intravenous immunoglobulin (adjusted odds ratio 1.21, 95% CI 0.40-3.70). There were no clinically and statistically significant differences in secondary outcomes. The one serious adverse event in the placebo group and the three in the methotrexate group were not thought to be related to treatment. INTERPRETATION: Oral methotrexate 15 mg weekly showed no significant benefit, but limitations in the trial design and the high rate of response in the placebo group meant that a treatment effect could not be excluded. This study can inform design of future trials in CIDP. FUNDING: The GBS/CIDP Foundation International. PMID- 19136304 TI - Long-term follow up (37-69 years) of patients with bladder exstrophy treated with ureterosigmoidostomy: uro-nephrological outcome. AB - OBJECTIVE: To describe the urological and nephrological long-term outcome of patients born with classical bladder exstrophy treated with bilateral ureterosigmoidostomies in early childhood. PATIENTS AND METHOD: Out of 42 patients born with bladder exstrophy in Switzerland between 1937 and 1968, 25 participated in this study; seven had died, seven were lost to follow up and three refused consent. Assessment included chart review, clinical examination, and assessment of renal function and morphology. RESULTS: After a follow-up period of 37-69 years ((mean 50 years), 13 of the 25 participants (52%) had their ureterosigmoidostomy still in place. All others had different forms of urinary diversions. Fifteen (60%) patients had normal renal function or mild chronic kidney disease as assessed by estimated glomerular filtration rate. Three patients were on renal replacement therapy. MRI (n=16) showed 10 morphologically normal kidneys. One patient suffered from adenocarcinoma of the colon, five had benign colonic polyps, one urethral papillary carcinoma and 18 no evidence of tumor. CONCLUSION: The majority of our patients have normal or mildly impaired renal function and a well functioning ureterosigmoidostomy. This is remarkable, given the fact that ureterosigmoidostomies are considered to be refluxing high pressure reservoirs at risk of renal injury and malignancy. PMID- 19136305 TI - Synovial B cells of rheumatoid arthritis express ZAP-70 which increases the survival and correlates with the inflammatory and autoimmune phenotype. AB - B cells have acquired an important role in the pathogenesis of rheumatoid arthritis (RA) since B cell depletion allowed to rescue patients poorly responders to TNFalpha blockers. This study focused on the involvement of ZAP-70 as a bio-marker of B cells immune activation in RA. ZAP-70 expression in synovial fluid (SF) B cells obtained from RA patients was increased compared to SF B cells of osteoarthritis (OA) patients. Moreover we found that ZAP-70 positive/CD38 positive and ZAP-70 positive/CD5 positive B cells were enriched in SF. The analysis of B cell apoptosis in vitro showed that the percentage of ZAP-70 negative B cells spontaneously undergoing apoptosis was significantly higher than ZAP-70 positive B cells. The ZAP-70 positive B cell ratio (SF/peripheral blood (PB)) showed a positive correlation with SF autoantibody levels and with local levels of BAFF and IL6. ZAP-70 positive B cells seem to define a subset characterized by increased survival and high relationship with local inflammation and autoimmunity. PMID- 19136306 TI - Endoluminal procedures for bariatric patients: expectations among bariatric surgeons. AB - BACKGROUND: Primary and revisional bariatric endoluminal procedures are currently being developed. Acceptable levels of risk and weight loss for these procedures have not yet been established. The aim of this study was to evaluate the expectations and concerns among bariatric surgeons regarding these procedures. METHODS: The American Society for Metabolic and Bariatric Surgery Emerging Technologies Committee developed a questionnaire that was distributed to the membership. Risk tolerance was assessed with comparison to commonly performed endoscopic and bariatric procedures. The percentage of excess weight loss (EWL) ranges were provided to assess the expectations for results 1 year after the procedure. RESULTS: A total of 214 responses were returned. The acceptable level of risk to achieve 10-20% EWL after primary and revisional procedures was equivalent to, or less than, that of a therapeutic endoscopic procedure for 81% and 76% of respondents, respectively. The acceptable level of risk to achieve 30 40% EWL after primary and revisional procedures was equivalent to that after laparoscopic adjustable gastric banding for 45% and 35% of respondents, respectively and equivalent to that after laparoscopic Roux-en-Y gastric bypass for 8% and 22%, respectively. In addition, 62% of respondents responded that 10 30% EWL would be acceptable for revisional procedures, and 35% responded that 10 30% EWL would be acceptable after a primary procedure. The primary concern was unproven efficacy, followed by durability, poor weight loss, availability of equipment, and procedural risk. Finally, 58% would not be willing to recommend an endoluminal procedure until the efficacy has been established, regardless of the risk. CONCLUSION: Risk tolerance and weight loss expectations among bariatric surgeons are different for primary and revisional endoscopic procedures. Most surgeons were unwilling to consider endoluminal procedures for their patients until the efficacy has been proven. PMID- 19136307 TI - Failure of adjustable gastric banding: starting BMI of 46 kg/m2 is a fulcrum of success and failure. AB - BACKGROUND: To determine the body mass index (BMI) located at the fulcrum of success and failure in a prospective study conducted at the University of Texas Health Science Center at Houston. On average, our patients whose percentage of excess weight loss (%EWL) was >50% at 1 year had a significantly lower BMI than those with <30% EWL. METHODS: We prospectively collected the weight loss data for 430 patients who had had an adjustable gastric band placed. We stratified the %EWL within 1 year for patients with a BMI of 30-59 kg/m2. A line was generated for the %EWL over time for BMI groups of 30-39, 40-49, and 50-59 kg/m(2) and compared with the average %EWL over time. The y-intercepts of the resulting four lines were graphed against the average BMI for each group. RESULTS: The generated y-intercept line had an R2 of .9237. Using the equation of this line and the known y-intercept for the average, we solved for x, resulting in a BMI of 46 kg/m2. Patients with a BMI <46 kg/m2 had a 50% EWL at 1 year, and those with a BMI >46 kg/m2 had only a 33% EWL at 1 year. The %EWL between the groups was significantly different at all measured intervals (P <.0001). CONCLUSION: A BMI of 46 kg/m2 identifies those at high risk of failure to lose a significant percentage of excess weight after adjustable gastric banding and who require closer follow-up. Furthermore, patients who have a BMI >46 kg/m2 should be advised that their weight loss might be suboptimal at 1 year. PMID- 19136308 TI - Early jejunojejunostomy obstruction after laparoscopic gastric bypass: case series and treatment algorithm. AB - BACKGROUND: To review our experience with early jejunojejunostomy obstruction (JJO) at a large academic teaching hospital and provide a management algorithm. Early JJO is a known and often overlooked complication of laparoscopic Roux-en-Y gastric bypass. METHODS: From 2003 to 2007, 1097 patients underwent laparoscopic Roux-en-Y gastric bypass at our institution. Data, including patient demographics, co-morbidities, intraoperative data, peri- and postoperative complications, and outcomes, were prospectively recorded and retrospectively reviewed. RESULTS: Early post-laparoscopic Roux-en-Y gastric bypass JJO occurred in 13 patients (1.2%). The average time to presentation was 15 days (range 5-27). Patients presented with a combination of nausea, vomiting, and abdominal pain; all underwent computed tomography to confirm the diagnosis. The causes of JJO included dietary noncompliance (46%), anastomotic edema (23%), narrowing of the jejunojejunostomy at surgery (23%), and luminal clot (8%). Management was determined using our proposed treatment algorithm. Three patients (23%) required operative intervention, with the remainder successfully treated conservatively. CONCLUSION: From our experience, we propose a treatment algorithm for standardized management of early JJO, reserving reoperation for those who are acutely ill on presentation or those in whom conservative management fails. A review of our series using this algorithm has suggested that most patients can be successfully treated nonoperatively; however, bariatric surgeons must maintain a low threshold for surgical re-intervention in cases in which rapid recovery is not seen. PMID- 19136310 TI - Obesity is associated with increased prevalence and severity of pelvic floor disorders in women considering bariatric surgery. AB - BACKGROUND: Although an association between obesity and urinary incontinence (UI) has been reported, the association between obesity and other PFDs is less clear. The aim of this study was to determine the prevalence of pelvic floor disorders (PFDs), including stress urinary incontinence (SUI), urge urinary incontinence (UUI), pelvic organ prolapse (POP), and anal incontinence (AI), in obese women contemplating bariatric surgery compared with nonobese subjects at a tertiary care referral hospital. METHODS: From September 2006 to December 2007, obese women contemplating bariatric surgery and nonobese women from general gynecology clinic completed a validated screening questionnaire for PFDs, the Sandvik urinary incontinence severity index, and the Rockwood fecal incontinence severity index. RESULTS: A total of 217 obese (mean body mass index of 50 +/- 10 kg/m(2)) and 210 nonobese controls (mean body mass index 23 +/- 3 kg/m(2)) were screened. The presence of any PFD occurred in 159 patients (75%) in the obese group compared with 89 nonobese patients (44%; P <.0001). More obese patients experienced SUI, UUI, and AI, but not POP. Obese patients also had more severe UI and AI. Obesity remained a significant risk factor for UI and AI, even after adjusting for baseline differences in demographics and medical conditions, with an adjusted odds ratio of 4.1 (95% confidence interval 2.3-7.8) and 2.1 (95% confidence interval 1.1-4.1), respectively. CONCLUSION: The prevalence of PFDs, including SUI, UUI, and all forms of AI, was greater in the obese and morbidly obese women contemplating bariatric surgery. Obesity was also associated with an increased severity of UI and AI. Obesity appears to confer a fourfold and twofold increased risk of UI and AI, respectively. PMID- 19136311 TI - Silastic ring vertical gastric bypass: cohort study with 83% rate of 5-year follow-up. AB - BACKGROUND: Silastic ring vertical gastric bypass (SRVGB) with jejunal interposition is our standard operation for morbidly obese patients. We present the results of 5 years of follow-up in a cohort of patients who underwent SRVGB in 2001. METHODS: The records of all 160 consecutive patients who underwent SRVGB from January to December 2001 were reviewed. Of the 160 procedures, 143 were primary open cases, 14 were revisions from restrictive procedures, and 3 were laparoscopic cases. At 5 years, the body mass index and percentage of excess weight loss was available for 133 patients (83%) at office visits (n = 91, 68.4%), by telephone (n = 40, 30.1%), or by e-mail (n = 2, 1.5%). RESULTS: Of the 160 patients, 121 were women and 39 were men, with a mean age of 33.15 +/- 10.0 years, percentage of ideal body weight of 195.7% +/- 40.8%, and body mass index of 44.6 +/- 9.3 kg/m(2). The mean hospital stay was 3 +/- 1 days. One patient (.6%) died of a pulmonary embolus. Early complications included 3 cases (1.87%) of upper gastrointestinal bleeding and 4 gastric leaks (2.5%): 2 (1.36%) from primary cases and 2 (14.29%) from revisional cases. Late complications included 32 patients (20%) with incisional hernias, 20 (12.5%) with anemia, 14 (8.8%) with dumping, 4 (2.5%) with gastrojejunal stricture, 2 (1.25%) with intestinal obstruction, and 2 (1.25%) requiring silastic ring surgical removal. The 5-year follow-up data were available for 133 patients (83%). The mean body mass index in this group was 27 +/- 5 kg/m(2), with a percentage of excess weight loss of 83% +/- 18.3% at 5 years postoperatively. CONCLUSION: The results of our study have shown that SRVGB is an effective operation for promoting lasting weight loss, with acceptable mortality and complication rates. PMID- 19136312 TI - Revisional operations for marginal ulcer after Roux-en-Y gastric bypass. AB - BACKGROUND: Marginal ulcer is a potentially serious complication of Roux-en-Y gastric bypass (RYGB). This study reviewed 1 surgeon's experience with 39 revisional operations for intractable marginal ulcer after primary RYGB. METHODS: A total of 2282 consecutive patients underwent RYGB by 1 surgeon from 1984 to 2006, of which 1621 were open and 661 laparoscopic. The stomach was transected in laparoscopic RYGB and was left undivided in the open group. All revisions included ulcer excision, revision of the gastrojejunostomy with gastric transection as needed. Six patients underwent vagotomy. RESULTS: Of the 2282 patients, 122 (5.3%) developed marginal ulcers (88 and 34 from the open and laparoscopic group, respectively). Of these 122 patients, 39 (32%) underwent revision for intractability (35 open and 4 laparoscopic). Of these 39 patients, 28 (71.7%) had gastrogastric fistulas. Risk factors for ulcer (medication, smoking) were present in 26 patients (66.6%). The primary indications of intractability included abdominal pain (66.6%), gastrointestinal bleeding (20.5%), stomal obstruction (10.2%), and perforation (2.5%). Early postoperative complications included 2 leaks (5.1%) and 2 wound infections (5.1%). Three smokers (7.7%) developed recurrent ulcers postoperatively. One patient died of acute respiratory distress syndrome 5 months postoperatively. Of the 39 patients, 34 (87%) remained asymptomatic after revision. The revision rate was significantly less after laparoscopic RYGB (.6%) than after open RYGB (2.1%; P < or =.0025). CONCLUSION: The results of our study have shown that operations for intractable marginal ulcer after RYGB are highly successful in nonsmokers. Patients who undergo laparoscopic RYGB with gastric transection are less likely to require revision than patients who undergo RYGB with an incontinuity gastric partition. The rate of reoperation for marginal ulcer was greater than anticipated. PMID- 19136313 TI - Clinical and radiologic findings of massive gastric prolapse after laparoscopic adjustable gastric banding. PMID- 19136314 TI - Gastric banding with simultaneous panniculectomy: two case reports on technique. PMID- 19136316 TI - Factors affecting rejection of bariatric patients from an academic weight loss program. AB - BACKGROUND: To determine the factors affecting rejection of bariatric candidates at an accredited, American College of Surgeons Level 1A, bariatric program. Bariatric surgery "Centers of Excellence" use a multidisciplinary team to screen patients for eligibility for surgery using insurance, medical history, psychological evaluation findings, and the surgeon assessment. Few studies have reported on the frequency or reasons for patients not being accepted for surgery among high-volume academic bariatric programs. METHODS: From March to September 2007, 299 consecutive patients were accepted for evaluation into an accredited bariatric program and tracked for the incidence of rejection for weight loss surgery. The primary reasons for rejection included a lack of insurance coverage, being medically unfit, psychological or social inappropriateness, and a body mass index (BMI) that did not meet the cutoff (BMI<35 kg/m2 or <40 kg/m2 without co morbid conditions). RESULTS: Of 299 screened patients, 90 (30.1%) were not accepted for surgery by the multidisciplinary team. The most frequent reason was the lack of insurance coverage (47.8%). Primary care physicians were the most common source of patient referral. All but 1 of the patients excluded because of an inadequate BMI (n=13) had been referred by friends, co-workers, or themselves from information received from the Internet or television. CONCLUSION: Approximately one third of screened patients were not accepted for surgery by an academic bariatric program. Self- or social referral appeared to correlate with rejection because the BMI did not meet the criteria for surgery. This suggests inadequate information among social referral networks and/or in the media. Long term follow-up will determine the health outcomes of patients not cleared for weight loss surgery. PMID- 19136315 TI - A rodent model of metabolic surgery for study of type 2 diabetes and positron emission tomography scanning of beta cell mass. AB - BACKGROUND: Type 2 diabetes mellitus is a worldwide healthcare problem with major socioeconomic implications. Metabolic surgical procedures have been shown to improve diabetes, but the mechanism of action is poorly understood. The Goto Kakizaki (GK) rodent is a type 2 diabetic animal model that is ideally situated for studying the effect of surgery on diabetes; however, the operative mortality is high. The aim of this study was to describe the operative technique, improvements in perioperative management, and the technique of micro-positron emission tomography (PET) scanning of the beta-cell mass in GK rodents. METHODS: A total of 53 GK rats were divided into 1 of 3 operative groups: sham, sleeve gastrectomy, and duodenojejunal bypass. A subset of animals underwent micro-PET scanning with [11C]-dihydrotetrabenazine to determine the vesicular monoamine transporter 2 binding index, an indicator of beta-cell mass. RESULTS: The 30-day mortality in the sham and sleeve gastrectomy rodents was 0; however, 2 sleeve gastrectomy rodents developed enterocutaneous fistula and 1 developed an abscess. In the duodenojejunal bypass group, the initial mortality rate was close to 90%; however, refinements in the surgical technique and perioperative management (fluids, antibiotics, pain control) lowered the mortality rate to 60%. The surgical technique is discussed in detail. [11C]-Dihydrotetrabenazine uptake in the pancreas was demonstrated on micro-PET scanning in the sham and duodenojejunal bypass rodents. CONCLUSION: Intensive medical management in the perioperative period and attention to the operative technique lowered the mortality. [11C]-Dihydrotetrabenazine micro-PET scanning is a feasible method for assessing the beta-cell mass in GK rodents and could prove to be an important modality for evaluating beta-cell performance in type 2 diabetes. PMID- 19136317 TI - Differential effects of hypoxia and acidosis on p53 expression, repair of UVC damaged DNA and viability after UVC in normal and tumor-derived human cells. AB - Hypoxia and low pH are commonly associated with the tumor microenvironment. We have examined the effects of hypoxia alone (HA) and hypoxia coupled to low pH (HApH) on p53 expression, nucleotide excision repair (NER) and cellular sensitivity to UVC in normal human fibroblasts and human tumor cells. p53 expression was measured using Western blotting, NER using host cell reactivation (HCR) of a UV-damaged reporter gene and cell sensitivity using the MTT assay. HApH resulted in a transient increase in p53 expression in normal fibroblasts at 6h and in tumor cells at 6-18h. In normal fibroblasts HApH resulted in a transient increase in HCR at early times (12-24h) and a concomitant decrease in UVC sensitivity. In contrast, for the tumor-derived cells, the increased HCR of the UVC-treated reporter gene was delayed (36-40h) and UVC sensitivity increased or remained the same after HApH treatment. These results suggest that early upregulation of p53 and increased repair of UV-damaged DNA after HApH treatment is required for increased cell viability after UVC. HA treatment alone also resulted in a transient increase in HCR of the UVC-damaged reporter gene at early times (12-24h) in normal fibroblasts and a delayed increase (36-40h) in the tumor derived cells. However, the enhanced p53 expression was less or even absent for treatment with HA alone, and HA had no significant affect on cell viability after UVC for any of the cell lines. These results indicate a different cellular response following HApH compared to HA alone. PMID- 19136318 TI - Shape memory polymer foams for cerebral aneurysm reparation: effects of plasma sterilization on physical properties and cytocompatibility. AB - Shape memory polyurethanes (SMPUs) represent promising candidate materials for aneurysm embolization, since they could enable clinical problems still associated with these clinical procedures to be overcome. In this work, we report on the characterization of physicochemical, thermomechanical and in vitro interface properties of two SMPU foams (Cold Hibernated Elastic Memory, CHEM), proposed as a material for embolization devices in minimally invasive procedures. Moreover, because device sterilization is mandatory for in vivo applications, effects on the properties of the foams after plasma sterilization were also evaluated. Both foams (CHEM 3520 and CHEM 5520) showed excellent shape recovery ability (recovery rate, R(r), up to 99%) in conventional shape recovery tests, performed at constant heating rate. Transition temperatures (T(trans)), determined by tandelta peaks in dynamic mechanical analysis (DMA), were 32.2 and 45.1 degrees C, for CHEM 3520 and 5520, respectively. The value of T(trans) affects shape memory ability in the recovery test at 37 degrees C, which simulates the behavior after implantation of the device: in fact, R(r) was significantly higher for lower T(trans) foam (R(r) approximately 82% and R(r) approximately 46%, respectively, for CHEM 3520 and CHEM 5520). After plasma sterilization performed by a Sterrad sterilization system, an increase in open porosity was observed: this is probably due to the sterilization cycle; however, no effects on shape recovery behavior were observed. Furthermore, plasma treatment had no significant effect on L929 cells in in vitro cytotoxicity tests, performed on cell culture medium extracts in contact with foams for up to 7 days. Moreover, direct cytocompatibility tests showed a good colonization and growth from L929 cells on CHEM foams, suggesting the effectiveness of an in vivo healing process. All these results seem to suggest that CHEM foams could be advantageously used for manufacturing devices for mini-invasive embolization procedures of aneurysms. PMID- 19136319 TI - The behavior of rat tooth germ cells on poly(vinyl alcohol). AB - The purpose of this study was to evaluate the behaviors of rat tooth germ (TG) cells cultured on poly(vinyl alcohol) (PVA). It was found that TG cells suspended and aggregated to form three-dimensional spheroids on PVA. Compared with traditional monolayered cells on tissue culture polystyrene, TG cell spheroids on PVA obviously increased the alkaline phosphatase activity, the degree of mineralization, and upregulated both osteopontin and dentin matrix protein 1 genes, regardless of the seeding density. Surprisingly, PVA appears to activate the alkaline phosphatase activity and mineralization effects on TG cell spheroids in the absence of a differentiation medium. Furthermore, the present study indicates that integrins may play an important role in the mineralization on TG cell spheroids by adding Arg-Gly-Asp (RGD) peptides. Therefore, the information presented here should help to clarify the role of PVA in the regulation of the mineralization, differentiation and integrin-mediation of TG cells. PMID- 19136320 TI - Degradable particulate composite reinforced with nanofibres for biomedical applications. AB - Nanofibre-based structures and their composites are increasingly being studied for many biomedical applications, including tissue engineering scaffolds. These materials enable architectures resembling the extracellular matrix to be obtained. The search for optimized supports and carriers of cells is still a major challenge for the tissue engineering field. The main purpose of this work is to develop a novel composite structure that combines microparticles and nanofibres in reinforced polymeric microfibres. This innovative combination of materials is obtained by melting extrusion of a particulate composite reinforced with chitosan nanofibre meshes (0.05 wt.%) produced by the electrospinning technique. The reinforced microfibres were analysed by scanning electron microscopy and showed a considerable alignment of the chitosan nanofibres along the longitudinal main axis of the microfibre composite structure. The tensile mechanical properties revealed that the introduction of the nanofibre reinforcement in the particulate microfibre composite increased the tensile modulus by up to 70%. The various structures were subjected to swelling and degradation tests immersed in an isotonic saline solution at 37 degrees C. The presence of chitosan nanofibres in the particulate microfibres enhances the water uptake by up to 24%. The combination of good mechanical properties and enhanced degradability of the developed structures is believed to have great potential for various biomedical applications, including three-dimensional fibre mesh scaffolds to be applied in the field of bone tissue engineering. PMID- 19136321 TI - Intramembranous bone tissue response to biodegradable octacalcium phosphate implant. AB - Previous studies showed that synthetic octacalcium phosphate (OCP) enhances bone formation coupled with its own osteoclastic biodegradation more than non biodegradable hydroxyapatite (HA), including sintered HA ceramic, when implanted in animal bone defects. The present study was designed to investigate whether synthetic OCP in granule form has biodegradable characteristics when implanted in the subperiosteal area of mouse calvaria in comparison with non-sintered stoichiometric HA, especially in relatively short periods after implantation. OCP crystals exhibited plate-like morphology, whereas HA crystals had a sphere-like structure. Both crystals had large pore volumes >75% in total, with micropores within the granules. Direct bonding of newly formed bone was discernible in HA until 35 days after implantation by element analysis for calcium and phosphorus. However, histomorphometric analysis demonstrated that bone formation was facilitated on OCP surfaces with greater alkaline phosphatase activity than on HA up to 21 days. The surfaces attacked by tartrate-resistant acid phosphatase positive osteoclast-like cells were significantly greater than those of HA. OCP became encapsulated and replaced with new bone with prolonged implantation periods up to 180 days. The results suggest that the enhanced bone formation in mouse calvaria could be associated with the biodegradable nature of OCP, and that OCP could be used in augmenting intramembranous bone volume. PMID- 19136322 TI - Reconstruction of the first and second metatarsals with free vascularised double barrelled fibular graft after resection of a chondrosarcoma. AB - We report the case of a 71-year-old woman with a chondrosarcoma originating in the first metatarsal of the left foot. We performed a wide resection and reconstructed the first and second metatarsals with a free vascularised double barrelled fibular graft. Her limb was successfully salvaged with functional and aesthetic results. Two years after surgery, the patient is able to walk without any ortho-prosthetic devices and has returned to farm work. The details of the technique applied in this case have been described in this article. PMID- 19136323 TI - Sentinel bruising in metastatic malignant melanoma. PMID- 19136324 TI - A useful technique to obtain adequate negative pressure for liposuction. PMID- 19136325 TI - Improved noninvasive coronary angiography in morbidly obese patients with dual source computed tomography. AB - BACKGROUND: Morbidly obese persons (body mass index [BMI; in kg/m(2)] >or= 40) have an increased risk of cardiovascular morbidity and mortality but have reduced accuracy with conventional cardiac testing and coronary CT angiography (CCTA). OBJECTIVE: This study investigated a novel dual-source computed tomography (DSCT) acquisition and reconstruction method for coronary imaging in morbidly obese patients. METHODS: This was a observational study in which each patient served as his or her own control. After a single DSCT acquisition using a novel method, standard quarter-scan image reconstructions at a temporal resolution of 83 milliseconds were compared with temporal resolution reconstructions at 105, 125, and 165 milliseconds. Images were evaluated for diagnostic adequacy score and for image noise, signal-to-noise ratio, and contrast-to-noise ratio. In each patient, the image reconstruction with the best visual diagnostic score was compared with the control image for quantitative measures. RESULTS: Fifty patients (32 female; mean +/- SD age, 51 +/- 10 y; mean BMI, 44.8 +/- 5.6) were enrolled. Scans were of diagnostic quality in 47 (94%) patients using the "best reconstruction" compared with 38 (76%) patients using quarter-scan reconstruction. Significant improvements were observed in noise (42 +/- 16 HU versus 56 +/- 19 HU; P < 0.0001), contrast-to-noise ratio (8.4 +/- 3.3 HU versus 7.0 +/- 2.2 HU; P = 0.0038), and signal-to-noise ratio (7.6 +/- 2.9 HU versus 6.5 +/- 3.5 HU; P = 0.030). CONCLUSIONS: CCTA with DSCT using a modified scan protocol and adjustable temporal reconstructions provides diagnostic image quality in >90% of morbidly obese patients. PMID- 19136326 TI - A comparison of the theory of unpleasant symptoms and the conceptual model of chemotherapy-related changes in cognitive function. AB - PURPOSE/OBJECTIVES: To explore the use of the Theory of Unpleasant Symptoms (TUS) as a model for describing the symptom experience related to the cognitive impairment associated with standard-dose chemotherapy and compare and contrast that use of the TUS with the Conceptual Model of Chemotherapy-Related Changes in Cognitive Function. DATA SOURCES: PubMed database. DATA SYNTHESIS: Retrospective and prospective clinical trials have substantiated the incidence of mild to moderate cognitive impairment. Patients have reported a significant effect on quality of life. Associated factors may include age, anemia, fatigue, depression, anxiety, hormone levels, cytokine release, and genetic makeup. Establishing the appropriateness of a working model to describe the relationships would provide additional structure and focus for empirical research. CONCLUSIONS: Both models have utility for describing the relationships of factors associated with chemotherapy-related cognitive impairment. IMPLICATIONS FOR NURSING: Blending of the TUS and the Conceptual Model of Chemotherapy-Related Changes in Cognitive Function may provide an enhanced framework for further research about the physiologic and psychological aspects of chemotherapy-related cognitive impairment. PMID- 19136327 TI - Application of multilevel growth-curve analysis in cancer treatment toxicities: the exemplar of oral mucositis and pain. AB - PURPOSE/OBJECTIVES: To introduce the use of a statistical technique known as multilevel growth-curve analysis and illustrate how the method can be advantageous in comparison with traditional repeated measures for the study of trajectories of signs and symptoms in individual patients over time. DATA SOURCES: Data were derived from use of the technique in a randomized clinical trial of a psychoeducational intervention to reduce severity of oral mucositis and oral pain. DATA SYNTHESIS: The development of new biologic models that seek to explain clustering of signs and symptoms or the appearance and resolution of signs and symptoms motivates the need to use more sophisticated statistical techniques to test such models. CONCLUSIONS: The application of multilevel growth model to an existing data set demonstrates that the model can be effective in the study of individual differences in trajectories of change in signs and symptoms. IMPLICATIONS FOR NURSING: This method for the study of changes in patients' signs and symptoms over time can be of particular interest to nursing, both from a clinical point of view and as a way to test theoretical models that have been proposed to capture patient experiences with signs and symptoms. PMID- 19136328 TI - A model linking uncertainty, post-traumatic stress, and health behaviors in childhood cancer survivors. AB - PURPOSE/OBJECTIVES: To consolidate the literature and provide a model to explain the links among uncertainty, post-traumatic stress syndrome, and health behaviors in adolescent and young adult childhood cancer survivors. DATA SOURCES: A systemic review of related literature and theory was used for the proposed model. The literature pertaining to the Uncertainty in Illness Theory, childhood cancer late effects, post-traumatic stress, and health behaviors was reviewed and critiqued from three data sets from 1979-2007: MEDLINE, PsycInfo, and CINAHL. Key words used for the search were uncertainty and post-traumatic stress as well as health behaviors, including smoking, alcohol use, unsafe sex, sunscreen use, and physical inactivity. DATA SYNTHESIS: Childhood cancer survivors living with chronic uncertainty may develop a new view of life and, as a result, adopt more health-promotion behaviors and engage in less health-risk behaviors. However, survivors living with chronic uncertainty may generate symptoms similar to post traumatic stress disorder and, therefore, adopt fewer health-promotion behaviors and engage in more health-risk behaviors. CONCLUSIONS: The uncertainty that pervades the childhood cancer experience can lead to the development of symptoms that resemble those of post-traumatic stress. The symptoms can interfere with the adoption of healthy lifestyle behaviors and avoidance of health-risk behaviors. IMPLICATIONS FOR NURSING: The theoretically derived model outlined in this article can be used to guide clinical interventions and additional research into the health behaviors of childhood cancer survivors. PMID- 19136329 TI - Spiritual needs of Taiwan's older patients with terminal cancer. AB - PURPOSE/OBJECTIVES: To explore the core constitutive patterns from the perspective of the older patient with terminal cancer, the major foci of spiritual needs, and professional actions that manifest tangibly as spiritual care. DESIGN: Hermeneutic inquiry. SETTING: Two leading teaching hospitals in Taiwan. SAMPLE: 35 older patients with terminal cancer with a life expectancy of three months. METHODS: Participatory observation and in-depth interviews were the major data collection strategies. Hermeneutic inquiry was used to disclose, document, and interpret participants' lived experiences. MAIN RESEARCH VARIABLES: The perceptions of older Taiwanese patients with terminal cancer and their particular healthcare needs across various transition stages. FINDINGS: Two constitutive patterns, "caring for the mortal body" and "transcending the worldly being," each with three themes and a related transformational process emerged from the data analysis. Two foci of spiritual needs were discovering patients' concerns about their dying bodies and their worldly being and facilitating the transcendence of the spiritual being from the physical realm. CONCLUSIONS: Caring for older Taiwanese patients with terminal cancer and helping them transcend the fear of the unknown when they give up worldly being can produce insights into patients' primary concerns, perceptions, and spiritual needs to help them achieve a "good" death. IMPLICATIONS FOR NURSING: The knowledge gained in this study and the six major spiritual care actions identified may empower oncology nurse educators to develop culturally valid spiritual care courses. Research-based guidance for nurse clinicians who are taking care of older Taiwanese patients with terminal cancer will be provided. PMID- 19136330 TI - Knowledge, attitudes, and behaviors of nursing and midwifery students regarding breast self-examination in Turkey. AB - PURPOSE/OBJECTIVES: To investigate knowledge, attitudes, and behaviors of nursing and midwifery students regarding breast self-examination (BSE). DESIGN: Descriptive, cross-sectional. SETTING: Aydin School of Health at Adnan Menderes University in Aydin, Turkey. SAMPLE: 244 female students of nursing and midwifery. METHODS: Data were collected with a questionnaire. MAIN RESEARCH VARIABLES: BSE-related knowledge, attitudes, and behaviors. FINDINGS: More than half of the study participants stated they had sufficient information about BSE from varied sources, primarily from school curricula. The students were knowledgeable about who should perform BSE and its recommended frequency; however, their knowledge of BSE techniques was limited. First-year students had negative attitudes about BSE but became more positive as they progressed in their education. Half of the study sample stated they performed BSE at recommended times and intervals, but only one-fifth used recommended BSE positions and techniques. The main reasons for not performing BSE included not knowing how to perform it (57%), not having any history of problems in the breast (39%), and forgetfulness (18%). CONCLUSIONS: The results demonstrate that nursing and midwifery education has a positive effect on students' knowledge, attitudes, and behaviors regarding BSE. IMPLICATIONS FOR NURSING: The findings suggest that nursing and midwifery students should be thoroughly prepared to perform BSE on themselves so they can educate other women about this important preventive procedure. PMID- 19136332 TI - Stand up for your profession. PMID- 19136335 TI - Building a collaborative nursing practice to promote patient education: an inpatient and outpatient partnership. PMID- 19136336 TI - Compassion fatigue: are you at risk? PMID- 19136337 TI - Predictors of fatigue 30 days after completing anthracycline plus taxane adjuvant chemotherapy for breast cancer. AB - PURPOSES/OBJECTIVES: To identify the predictors of fatigue 30 days after completing adjuvant chemotherapy for breast cancer and whether differences are observed between a behavioral sleep intervention and a healthy-eating attention control group in predicting fatigue. DESIGN: Descriptive, exploratory, secondary analysis of a randomized clinical trial. SETTING: Outpatient oncology patients in a midwestern U. S. city. SAMPLE: 96 women, ages 29-83 years, 72% married, 95% white, diagnosed with stage I-IIIA breast cancer, receiving adjuvant anthracycline and taxane chemotherapy. METHODS: Participants were randomized to a behavioral sleep intervention group or an attention control group. Participants completed data collection prior to and during the peak and rebound days of the initial chemotherapy treatment cycle and after the last treatment. MAIN RESEARCH VARIABLES: Fatigue, circadian rhythms of activity, objective and subjective sleep wake, and objective and subjective activity-rest. FINDINGS: Predictors of fatigue were less total sleep time prior to treatment, higher fatigue prior to treatment and at the peak, and less energy upon awakening on rebound days. In the control group, predictors of higher fatigue were higher fatigue prior to treatment, higher body mass index, higher number of positive lymph nodes, and less daytime dysfunction. For the intervention group, lower peak activity at the peak of initial treatment differentially predicted fatigue. CONCLUSIONS: Results suggest the sleep intervention group participants who maintained activity balanced with sleep at the peak of the initial treatment benefited most from the intervention. IMPLICATIONS FOR NURSING: Nurses should screen for fatigue prior to initial chemotherapy treatment and at regular intervals, further assess for poor sleep in patients who report fatigue of 4 or higher (on a 0-10 scale), and use evidence based guidelines to select appropriate interventions. PMID- 19136338 TI - Use of buffered lidocaine in bone marrow biopsies: a randomized, controlled trial. AB - PURPOSE/OBJECTIVES: To determine whether a difference exists in perceived pain during preprocedure anesthetic injection for bone marrow biopsy between buffered and unbuffered lidocaine, to determine whether pain levels change over time, and to investigate relationships between perceived pain scores and other variables. DESIGN: A double-blind, randomized, experimental, crossover design. SETTING: A large hospital in the midwestern region of the United States. SAMPLE: 48 patients undergoing bone marrow biopsy. METHODS: The patients served as their own controls for the bilateral procedure. A 100 mm visual analog scale measured pain. A demographic questionnaire gathered the between-subjects exploratory variables. MAIN RESEARCH VARIABLES: Perceived pain scores and type of lidocaine anesthetic solution (buffered versus unbuffered). FINDINGS: Participants reported significantly lower pain scores on the side anesthetized with buffered lidocaine compared with the side anesthetized with unbuffered lidocaine. Higher pain scores were reported on the treatment side for participants who had received more than two surgical procedures. Patients who were members of a minority group had higher mean pain scores than Caucasians on the control side. CONCLUSIONS: Buffered lidocaine is superior to unbuffered lidocaine as an anesthetic for bone marrow biopsy procedures. IMPLICATIONS FOR NURSING: Advanced practice nurses perform a significant number of bone marrow biopsies and aim to improve patient comfort during invasive procedures. Use of unbuffered lidocaine should be questioned. PMID- 19136339 TI - Dietary supplement use in adult cancer survivors. AB - PURPOSE/OBJECTIVES: To assess dietary supplement use and its association with demographic and health-related characteristics among cancer survivors and to investigate differences in supplement use patterns by cancer site. DESIGN: A cross-sectional survey. SETTING: Computer-assisted telephone survey. SAMPLE: 1,233 adult (ages 30-69) survivors participating in the Penn State Cancer Survivor Study who underwent an interviewer-administered questionnaire. METHODS: Descriptive statistics with multivariate logistic regression to determine demographic, disease, and health-related predictors of supplement use. MAIN RESEARCH VARIABLES: Use of dietary supplements and types of supplements taken. FINDINGS: Supplement use ranged from 50% among blood cancer survivors to 85% among melanoma skin cancer survivors, with an overall prevalence rate of 73%. Multivariate logistic regression revealed statistically significant associations (p values < 0.05) between supplement use and older age (>or= age 50), higher levels of education and physical activity, female gender, lower body mass index, and white ethnicity. CONCLUSIONS: Overall, a wide variety of supplements were reported, although multivitamins, calcium and vitamin D combinations, and antioxidant vitamin combinations were the most prevalent. Seventy-eight percent of supplement users took more than one supplement. IMPLICATIONS FOR NURSING: The findings support continued efforts by oncology nurses to identify the types of supplements cancer survivors are using. Nurses should caution against the use of individual supplements as well as combinations of different supplements containing nutrient quantities above recommended daily intake levels. Furthermore, oncology nurses and other healthcare professionals should be receptive to questions and prepared to initiate conversations with patients about their use of dietary supplements. PMID- 19136340 TI - Where the dying live: a systematic review of determinants of place of end-of-life cancer care. AB - PURPOSE/OBJECTIVES: To describe the determinants of place of end-of-life (EOL) care for patients with cancer. DATA SOURCES: A systematic literature review of primary research studies (1997-2007) was conducted. Studies that investigated place of EOL care or identified place of EOL care in relation to outcomes were examined, their critical quality was appraised, and references were mapped. DATA SYNTHESIS: Of the 735 articles identified, 39 (representing 33 studies) met inclusion criteria. Two main research designs emerged: large-scale epidemiologic reports and smaller descriptive studies. Findings suggest that factors related to the disease, the individual, and the care and social environment influence place of EOL care for patients with cancer. Social support, healthcare inputs (from services and programs and healthcare provider contact), and patient preferences were the most important factors. CONCLUSIONS: Most patients with terminal cancer prefer home palliation; however, most die in an institution. The reasons are complex, with various determinants influencing decisions regarding place of EOL care. IMPLICATIONS FOR NURSING: Findings may highlight evidence-based interventions to assist patients and families facing decisions regarding place of EOL care. A clearer understanding of factors that influence place of EOL care for patients with cancer could enhance healthcare policy and guide needs-based modifications of the healthcare system. PMID- 19136341 TI - Examining the impact of socioeconomic status and socioecologic stress on physical and mental health quality of life among breast cancer survivors. AB - PURPOSE/OBJECTIVES: To examine how physical and mental health quality of life (QOL) varies in relation to the socioeconomic status and ethnicity among breast cancer survivors; to determine key socioecologic factors influencing outcomes. DESIGN: Cross-sectional. SETTING: Participants were recruited from the California Cancer Surveillance Program, from hospital registries, and from community agencies in southern California. SAMPLE: 703 multiethnic population-based breast cancer survivors, including European, African, Latina, and Asian Americans. METHODS: Participants completed a mailed questionnaire or answered a telephone survey. To identify socioeconomic status and socioecologic stress, four measures were used: household income, education, job type, and the Life Stress Scale. MAIN RESEARCH VARIABLES: Physical and mental health QOL, socioeconomic status (income, education, and job type), and socioecologic stress. FINDINGS: After controlling for the demographic and medical information, health-related QOL was significantly correlated to socioeconomic status, such that higher socioeconomic status groups expressed better QOL. Ethnic variations existed in QOL according to socioeconomic status. Socioecologic stress was the most important factor influencing physical and mental health QOL. CONCLUSIONS: The findings provide additional evidence that low socioeconomic status and high socioecologic stress exacerbate negative QOL sequelae. IMPLICATIONS FOR NURSING: Practice and research implications include the need for greater attention to QOL outcomes among at-risk lower socioeconomic status survivors and the recognition of the unique contributions of socioeconomic status, socioecologic stress, and ethnicity on physical and mental health QOL. PMID- 19136342 TI - Factors influencing men undertaking active surveillance for the management of low risk prostate cancer. AB - PURPOSE/OBJECTIVES: To identify and describe decision-making influences on men who decide to manage their low-risk prostate cancer with active surveillance. RESEARCH APPROACH: Qualitative, semistructured interview. SETTING: The Prostate Centre at Vancouver General Hospital in Canada. PARTICIPANTS: 25 patients diagnosed with low-risk prostate cancer and on active surveillance. METHODOLOGIC APPROACH: An interpretative, descriptive, qualitative design. MAIN RESEARCH VARIABLES: Factors that influenced men's decisions to take up active surveillance. FINDINGS: The specialists' description of the prostate cancer was the most influential factor on men choosing active surveillance. Patients did not consider their prostate cancer to be life threatening and, in general, were relieved that no treatment was required. Avoiding treatment-related suffering and physical dysfunction and side effects such as impotence and incontinence was cited as the major reason to delay treatment. Few men actively sought treatment or health-promotion information following their treatment decision. Female partners played a supportive role in the decision. The need for active treatment if the cancer progressed was acknowledged. Patients were hopeful that new treatments would be available when and if they needed them. Being older and having comorbidities did not preclude the desire for future active treatment. Patients carried on with their lives as usual and did not report having any major distress related to being on active surveillance. CONCLUSIONS: The study findings indicate that men are strongly influenced by the treating specialist in taking up active surveillance and planning future active treatments. As such, most men relied on their specialists' recommendation and did not perceive the need for any adjunct therapy or support until the cancer required active treatment. INTERPRETATION: Oncology nurses should work collaborative-ly with specialists to ensure that men receive the information they need to make informed treatment decisions. PMID- 19136343 TI - Ipilimumab for advanced melanoma: a nursing perspective. AB - PURPOSE/OBJECTIVES: To discuss the response patterns and side effects related to ipilimumab, a new immunotherapeutic agent under investigation in the treatment of advanced melanoma and other malignancies. DATA SOURCES: Published articles, abstracts, research data, and clinical experience. DATA SYNTHESIS: Ipilimumab is a fully human monoclonal antibody that inhibits the activity of cytotoxic T lymphocyte antigen-4 (CTLA-4), a naturally immunosuppressive molecule. The most common side effects are immune mediated (e.g., inflammatory diarrhea, pruritus) and appear to occur as a direct result of CTLA-4 inhibition and enhanced immune system activation. Side effects generally are grade I or II and resolve with standard treatments. Most grade III or IV events are managed successfully after swift diagnosis and treatment with corticosteroids; steroid-refractory events resolve after treatment with infliximab or mycophenolate. CONCLUSIONS: The response patterns and side effects associated with ipilimumab therapy greatly differ from those common to other advanced melanoma therapies (e.g., chemotherapy, cytokines, vaccines). IMPLICATIONS FOR NURSING: Nurses have an important role in educating patients about the differences between anti-CTLA-4 therapy and chemotherapy. In addition, teaching patients to recognize ipilimumab's side effects and report them early can result in fast treatment to prevent symptom progression from grade I or II to III or IV. Communication between nurses and patients throughout the treatment process will help patients benefit maximally from the new therapeutic strategy. PMID- 19136346 TI - Introduction to special section on occupational therapy. PMID- 19136347 TI - Doing daily life: how occupational therapy can inform psychiatric rehabilitation practice. AB - TOPIC: This paper provides an overview of occupational therapy in the context of psychiatric rehabilitation and mental health recovery. PURPOSE: The paper delineates practical aspects of occupational therapy's involvement in the mental health field with a discussion of occupation and the elements of conceptual models that guide the practice of occupational therapy. SOURCES USED: CINAHL, Psych Info, Medline. CONCLUSION: Occupational therapy is a key discipline in the field of psychiatric rehabilitation and brings to the field a strong theoretical and knowledge base along with unique procedures and practices. It is important for the psychiatric rehabilitation field to learn from all disciplines, including occupational therapy. PMID- 19136344 TI - Capecitabine-based combination therapy for breast cancer: implications for nurses. AB - PURPOSE/OBJECTIVES: To review available data and implications for nurses of combination regimens containing capecitabine for metastatic breast cancer. DATA SOURCES: Peer-reviewed publications or abstracts from major oncology conferences and reviews of capecitabine focusing on nursing implications. DATA SYNTHESIS: Capecitabine has proven efficacy in combination with docetaxel and is under evaluation in the neoadjuvant, adjuvant, and metastatic settings in combination with several oral and IV chemotherapeutic and biologic agents. CONCLUSIONS: Capecitabine-containing regimens demonstrate high activity in a range of settings but typically have more complex safety profiles, dose-modification schemes, and scheduling requirements than monotherapy. IMPLICATIONS FOR NURSING: Patients need to be aware of a wider range of likely side effects and should understand that they have been prescribed combination therapy rather than more simple, single agent treatments because of its potential to improve outcome. PMID- 19136348 TI - Functional assessment and intervention in occupational therapy. AB - TOPIC: Occupational therapists consider the person, the environment and the occupation (personally meaningful activities) in the assessment and intervention process with the ultimate goal being occupational performance. PURPOSE: This paper describes functional assessment and intervention from the perspective of the occupational therapist with specific examples of assessment instruments and intervention approaches. A scenario of a person in recovery with a particular occupational performance goal is woven throughout the paper to apply the concepts of functional assessment and intervention to a specific situation. SOURCES USED: CINAHL and PubMed databases; Existing assessment manuals and research on these instruments. CONCLUSIONS: Occupational therapists are just one discipline of many within psychiatric rehabilitation. Multidisciplinary teams can be most effective when they work collaboratively and maximize the expertise of all contributors. By describing the occupational therapy process, providing a scenario, and offering suggestions for collaboration, it is hoped that this paper fosters dialogue among occupational therapists and others involved in psychiatric rehabilitation. PMID- 19136349 TI - The Canadian Occupational Performance Measure: a tool for recovery-based practice. AB - TOPIC: This paper discusses the Canadian Occupational Performance Measure (COPM) which is a tool that supports goal setting and the assessment of change with an individual participating in daily activity. PURPOSE: The purpose of the paper is to examine the Canadian Occupational Performance Measure as a potential tool for use in psychiatric rehabilitation and recovery-oriented services. The paper considers the theoretical orientation of the Canadian Occupational Performance Measure and describes the instrument's structure and properties. In addition, there is a delineation of the instrument's utility in mental health and an examination of its relationship to psychiatric rehabilitation and recovery principles. SOURCES USED: The paper uses existing research on the instrument, examines psychosocial rehabilitation literature, and reflects on consultation with local and international researchers/practitioners. CONCLUSIONS: The Canadian Occupational Performance Measure is designed to foster partnership between clients and practitioners and it encourages identification of occupationally focused issues and goals. This instrument promotes an agenda of participation, resumption of life roles, and inclusion in environments of choice. PMID- 19136350 TI - Time use among people with psychiatric disabilities: implications for practice. AB - TOPIC: This paper reviewed the current literature regarding time use among people with psychiatric disabilities. PURPOSE: The purpose was to investigate what characterizes time use, occupational balance and occupational patterns among people with psychiatric disabilities, how time use relates to health in this group, and how time use can be used in research designs and addressed in assessments and interventions. SOURCES: The databases CINAHL and PubMed were searched, by combinations of terms such as: time use, time geography, occupational balance, occupational pattern, assessment, intervention, occupational therapy, lifestyle redesign, well-being, and mental health. CONCLUSIONS: Time use for people with psychiatric disabilities is often restricted to sleeping, eating, caring for oneself, and performing quiet activities. The target group is at risk of being both over- and under-occupied, and occupational imbalance may be regarded as an expression of the disability, thus shaped by a misfit between the person's capacities and environmental opportunities and demands. Several time-use methods exist for the study of daily occupations in people with psychiatric disabilities, but no intervention based on time use was found. Principles for a time-use based intervention are discussed. However, such an intervention needs to be investigated for relevance and effectiveness in future research. PMID- 19136351 TI - An occupational and rehabilitation perspective for institutional practice. AB - OBJECTIVE: The article aims to provide an occupational perspective on the lives of people with a serious mental illness who have committed a criminal offense and are incarcerated in a secure environment. PURPOSE: The article focuses on ways that institutions fail to meet occupational needs of such persons and the challenges for mental health and psychiatric rehabilitation professionals, including occupational therapists, in providing psychiatric rehabilitation to facilitate community integration and participation. The concepts of occupational deprivation, occupational imbalance, habits and occupational enrichment provide useful theoretical constructs underpinning practice endeavors. SOURCES USED: Ovid using Medline, PsychINFO, CINAHL, OTDBase, and ProQuest. CONCLUSIONS: There is a priority for research to validate tools to assess outcomes of occupations in secure settings, and the use of these tools to focus on which rehabilitation practices are correlated with establishing positive outcomes after release. Research evidence is also needed that demonstrates that occupational enrichment can result in observable and measurable outcomes that mitigate the negative effects of incarceration and support successful community re-entry of persons with mental illnesses who are offenders. PMID- 19136352 TI - What does recovery mean for me? Perspectives of Canadian mental health consumers. AB - OBJECTIVE: The objective of this study was to explore the meaning of recovery from the perspectives of consumers receiving mental health services in Canada. METHODS: Sixty semi-structured interviews were conducted with 54 mental health consumers in Montreal, Quebec City and Waterloo-Guelph, Ontario. RESULTS: Two contrasting meanings of recovery emerged. The first definition strongly attached recovery to illness while the second definition linked recovery to self determination and taking responsibility for life. CONCLUSIONS: The prominence of biomedical definitions of recovery suggests the need to find common ground between these two perspectives, if conceptualizations of recovery are to include the views of consumers who routinely experience the mental health system. PMID- 19136353 TI - Taking culture seriously: ethnolinguistic community perspectives on mental health. AB - OBJECTIVE: Ethnolinguistic communities are underserved by mental health systems in immigrant-receiving, multicultural societies, but their perspectives are seldom elicited in mental health research or reform planning. This article helps fill this gap by presenting community perspectives on concepts of mental health, mental illness and mental health experiences with five ethnocultural communities (Latin American, Mandarin-speaking Chinese, Polish, Punjabi Sikh and Somali) in Ontario, Canada. METHODS: Data were collected from 21 focus groups as part of a large-scale, participatory action research project called Taking Culture Seriously in Community Mental Health. RESULTS: The analysis focuses on how mental health and mental illnesses are described, how mental health care is experienced and what recommendations community members provide to improve the mental health system. CONCLUSIONS: Study findings illustrate the importance of the social context of immigration and settlement in conceptualizing mental health and mental distress. We conclude that systemic changes are needed to formulate collaborative, community-based strategies for mental health promotion and interventions. PMID- 19136354 TI - Making art: a qualitative study of personal and group transformation in a community arts studio. AB - OBJECTIVE: The objectives of this ethnographic study were to understand the effect of an integrated arts studio on the lives of the participants including the impact of social isolation, stigma and discrimination, and to study the importance of consumer perspective in the design and implementation of the program. METHODS: Methods included the use of semi-structured in-depth interviews, participant-observation, journal keeping, and document review with twenty individuals. Ten participants self identified as having mental illnesses, and their diagnoses were varied in severity and description; the other ten participants reported no mental illnesses. RESULTS: A key finding of the study was that art making provided participants the opportunity to build new identities and roles, and that through engagement in mutually meaningful activity, in this case making art, a community of artists developed. Art was also seen as a bridge creating access to the larger community. CONCLUSIONS: These findings add further evidence for the power of art making as a practical strategy to affect the health of individuals living with psychiatric disabilities as well as the communities in which they reside. PMID- 19136355 TI - Using the recovery knowledge inventory (RKI) to assess the effectiveness of a consumer-led recovery training program for service providers. AB - OBJECTIVE: This Australian study was designed to assess the effectiveness of a consumer-led recovery training program. METHODS: A non-equivalent control group study design was used to assess changes in recovery knowledge and attitudes pre training, immediately post-training, and at 6 months post-training. RESULTS: Relative to the comparison group, those receiving training demonstrated significant gains in knowledge at follow-up. CONCLUSIONS: A consumer-led training program was able to improve provider knowledge of recovery based practice. While the RKI was developed in the USA, it proved to be a useful measure of change in an Australian sample. PMID- 19136356 TI - The implications of race and ethnicity for shared decision-making. AB - TOPIC: Shared Decision Making is an innovative health-care approach gaining ground as part of the shift toward patient-centered medicine. It involves collaboration and discussion between doctors and patients to arrive at mutually satisfying treatment decisions. It places a strong emphasis on the therapeutic alliance, empowering patients to play an active role in their own care. PURPOSE: In this article, the author argues that research and development vis-a-vis shared decision-making must take serious account of ethno-racial status as a mediating variable. This may be a powerful determinant of broad orientations toward medical decision-making. SOURCES USED: Medline was used to extract relevant papers which were analyzed and intertwined with the author's existing knowledge of relevant sociological and anthropological evidence and theory. CONCLUSIONS: Specific proposals are made for a research agenda ensuring that ethno-racial issues are considered as a departure point rather than an afterthought in shared decision making model development. PMID- 19136357 TI - Transitional employment and its effects in Finland. AB - OBJECTIVE: The aim of this study was to explore the extent and type of transitional employment (TE) work undertaken in clubhouses in Finland and to better understand the influence of transitional employment on the rehabilitation goals of clubhouse members. Secondarily, the survey examined how TE possibly led to independent work or participation in adult education. METHODS: A survey was conducted with members of all Finnish Fountain House clubhouses who had participated in a transitional employment program from the beginning of the year 2002. A second survey was also conducted with clubhouse directors. RESULTS: Study results provide evidence that TE-participation leads to an active lifestyle and increases social inclusion, but it does not lead to permanent employment. CONCLUSIONS: Both members and leaders reported that transitional employment periods helped to evaluate an individual's working ability, but did not lead to steady employment in the open labor market. PMID- 19136358 TI - My journey to wellness. PMID- 19136359 TI - How will certification affect the field of psychiatric rehabilitation? PMID- 19136361 TI - RECOMBFLOW: a scientific workflow environment for Intragenomic Gene Conversion analysis in bacterial genomes, including the pathogen Streptococcus pyogenes. AB - Intragenomic Gene Conversion (IGC) is important in the evolution of bacteria but has only been analysed computationally in a few strains of Escherichia coli. This paper describes a scientific workflow system, called RECOMBFLOW, that automates this complex procedure for the analysis of more than 400 bacterial genomes, with a median analysis time per genome of less than 5 minutes. Results show that IGC varies greatly, both between different species and among multiple genomes of the same species. We analyse for the first time the large variation of IGC in the pathogen Streptococcus pyogenes, and also in non-pathogenic bacteria. PMID- 19136362 TI - Analysis of protein phosphorylation site predictors with an independent dataset. AB - Protein phosphorylation plays a fundamental role in most of the cellular regulatory pathways. Experimental detection of protein phosphorylation sites is labour intensive and often limited by the availability and optimisation of enzymatic reactions. The in silico prediction of phosphorylation sites using protein's primary sequences may provide guidelines for further experimental consideration and interpretation of phosphoproteomic data. An array of such tools exists over the internet and provides the prediction for protein kinase families. We developed an independent dataset to compare the performances of these methods to provide scientists with a better understanding of which method to use for their research. PMID- 19136363 TI - A practical algorithm for multiplex PCR primer set selection. AB - Selecting the minimum primer set with multiple constraints is an effective method for a successful and economical Multiplex Polymerase Chain Reaction (MP-PCR) experiment. However, there is no suitable algorithm for solving the problem. In this paper, a mathematical model is presented for the minimum primer set selection problem with multiple constraints. By introducing a novel genetic operator, we developed a parthenogenetic algorithm MG-PGA to solve the model. Experimental results show that MG-PGA can not only find a small primer set, but can also satisfy multiple biological constraints. Therefore, MG-PGA is a practical solution for MP-PCR primer design. PMID- 19136364 TI - Finding Significantly Expressed genes from time-course expression profiles. AB - This paper proposes a statistical method for finding Significantly Expressed (SE) genes from time-course expression. SE genes are time-dependent while non-SE genes are time-independent. This method models time-dependent gene expression profiles by autoregressive equations plus Gaussian noises, and time-independent ones by Gaussian noises. The statistical F-testing is used to calculate the probability (p-value) that a profile is time-independent. Both a synthetic dataset and a biological dataset were employed to evaluate the performance of this method, measured by the False Discovery Rate (FDR) and the False Non-discovery Rate (FNR). Results show that the proposed method outperforms traditional methods. PMID- 19136365 TI - Stabbing balls and simplifying proteins. AB - We address the problem of stabbing a sequence of indexed balls B = {B1,B2, . . . , Bn} in R(3), where Bi (1 0.12). CONCLUSIONS: We found decreased repeated-element methylation after exposure to traffic particles. Whether decreased methylation mediates exposure-related health effects remains to be determined. PMID- 19136373 TI - Sepsis alters the megakaryocyte-platelet transcriptional axis resulting in granzyme B-mediated lymphotoxicity. AB - RATIONALE: Sepsis-related mortality results in part from immunodeficiency secondary to profound lymphoid apoptosis. The biological mechanisms responsible are not understood. OBJECTIVES: Because recent evidence shows that platelets are involved in microvascular inflammation and that they accumulate in lymphoid microvasculature in sepsis, we hypothesized a direct role for platelets in sepsis related lymphoid apoptosis. METHODS: We studied megakaryocytes and platelets from a murine-induced sepsis model, with validation in septic children, which showed induction of the cytotoxic serine protease granzyme B. MEASUREMENTS AND MAIN RESULTS: Platelets from septic mice induced marked apoptosis of healthy splenocytes ex vivo. Platelets from septic granzyme B null (-/-) mice showed no lymphotoxicity. CONCLUSIONS: Our findings establish a conceptual advance in sepsis: Septic megakaryocytes produce platelets with acutely altered mRNA profiles, and these platelets mediate lymphotoxicity via granzyme B. Given the contribution of lymphoid apoptosis to sepsis-related mortality, modulation of platelet granzyme B becomes an important new target for investigation and therapy. PMID- 19136374 TI - Acute lung injury is reduced in fat-1 mice endogenously synthesizing n-3 fatty acids. AB - RATIONALE: Acute lung injury (ALI) remains an important cause of mortality in intensive care units. Inflammation is controlled by cytokines and eicosanoids derived from the n-6 fatty acid (FA) arachidonic acid (AA). The n-3 FA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and mediators derived from EPA and DHA possess reduced inflammatory potency. OBJECTIVES: To determine whether the ability of fat-1 mice to endogenously convert n-6 to n-3 FA, and thus generate an increased ratio of n-3 to n-6 FA, impacts experimental ALI. METHODS: We investigated ALI induced by intratracheal instillation of endotoxin in fat-1 and wild-type (WT) mice, assessing leukocyte numbers, protein concentration, and prostaglandin and cytokine levels in bronchoalveolar lavage fluid, as well as free FA in plasma, and lung ventilator compliance. Body temperature and motor activity of mice--markers of sickness behavior--were also recorded. MEASUREMENTS AND MAIN RESULTS: In ALI, fat-1 mice exhibited significantly reduced leukocyte invasion, protein leakage, and macrophage inflammatory protein-2 and thromboxane B(2) levels in lavage fluid compared with WT mice. Free AA levels were increased in the plasma of WT mice in response to endotoxin, whereas EPA and DHA were increased in the fat-1 group. Ventilator compliance was significantly improved in fat-1 mice. Body temperature and motor activity were decreased in ALI. fat-1 Mice recovered body temperature and motor activity faster. CONCLUSIONS: fat-1 Mice exhibited reduced features of ALI and sickness behavior. Increasing the availability of n-3 FA may thus be beneficial in critically ill patients with ALI. PMID- 19136376 TI - Gene plasticity in colonic circular smooth muscle cells underlies motility dysfunction in a model of postinfective IBS. AB - The cellular mechanisms of motility dysfunction in postinfectious irritable bowel syndrome (PI-IBS) are not known. We used a rat model of neonatal inflammation to test the hypothesis that gene plasticity in colonic circular smooth muscle cells underlies motility dysfunction in PI-IBS. Mild/moderate or severe inflammation was induced in neonatal and adult rats. Experiments were performed in tissues obtained at 7 days (short term) and 6-8 wk (long term) after the induction of inflammation. Severe inflammation in neonatal rats induced persistent long-term smooth muscle hyperreactivity to acetylcholine (ACh), whereas that in adult rat caused smooth muscle hyporeactivity that showed partial recovery in the long term. Mild/moderate inflammation had no effect in neonatal rats, but it induced smooth muscle hyporeactivity to ACh in adult rats, which recovered fully in the long term. Smooth muscle hyperreactivity to ACh resulted in accelerated colonic transit and increase in defecation rate, whereas hyporeactivity had opposite effects. Smooth muscle hyperreactivity to ACh was associated with increase in transcription rate of key cell-signaling proteins of the excitation-contraction coupling alpha1C subunit of Cav1.2 (L-type) calcium channels, Galphaq, and 20-kDa myosin light chain (MLC20), whereas hyporeactivity was associated with their suppression. Inflammation in adult rats induced classical inflammatory response, which was absent in neonatal rats. Severe neonatal inflammation enhanced plasma norepinephrine and muscularis propria vasoactive intestinal polypeptide in the long term. We conclude that severe, but not mild/moderate, inflammation in a state of immature or impaired stress and immune response systems alters the transcription rate of key cell-signaling proteins of excitation-contraction coupling in colonic circular smooth muscle cells to enhance their contractility and accelerate colonic transit and defecation rate. PMID- 19136375 TI - Fibrinogen genes modify the fibrinogen response to ambient particulate matter. AB - RATIONALE: Ambient particulate matter has been associated with systemic inflammation indicated by blood markers such as fibrinogen, implicated in promoting atherothrombosis. OBJECTIVES: This study evaluated whether single nucleotide polymorphisms (SNPs) within the fibrinogen genes modified the relationship between ambient particles and plasma fibrinogen. METHODS: In 854 myocardial infarction survivors from five European cities plasma fibrinogen levels were determined repeatedly (n = 5,082). City-specific analyses were conducted to assess the impact of particulate matter on fibrinogen levels, applying additive mixed models adjusting for patient characteristics, time trend, and weather. City-specific estimates were pooled by meta-analysis methodology. MEASUREMENTS AND MAIN RESULTS: Seven SNPs in the FGA and FGB genes shown to be associated with differences in fibrinogen levels were selected. Promoter SNPs within FGA and FGB were associated with modifications of the relationship between 5-day averages of particulate matter with an aerodynamic diameter below 10 microm (PM(10)) and plasma fibrinogen levels. The PM(10)-fibrinogen relationship for subjects with the homozygous minor allele genotype of FGB rs1800790 compared with subjects homozygous for the major allele was eightfold higher (P value for the interaction, 0.037). CONCLUSIONS: The data suggest that susceptibility to ambient particulate matter may be partly genetically determined by polymorphisms that alter early physiological responses such as transcription of fibrinogen. Subjects with variants of these frequent SNPs may have increased risks not only due to constitutionally higher fibrinogen concentrations, but also due to an augmented response to environmental inflammatory stimuli such as ambient particulate matter. PMID- 19136377 TI - A synthetic farnesoid X receptor (FXR) agonist promotes cholesterol lowering in models of dyslipidemia. AB - The nuclear hormone receptor farnesoid X receptor (FXR) plays a critical role in the regulation of bile acid, triglyceride (TG), and cholesterol homeostasis. WAY 362450 (FXR-450/XL335) is a potent synthetic FXR agonist as characterized in luciferase reporter assays and in mediating FXR target gene regulation in primary human and immortalized mouse hepatocytes. In vivo, WAY-362450 dose dependently decreased serum TG levels after 7 days of oral dosing in western diet-fed low density lipoprotein receptor-/- mice and in the diabetic mouse strains KK-Ay and db/db comparable to that achieved with the peroxisome proliferator activated receptor-alpha agonist, fenofibrate. WAY-362450 treatment also reduced serum cholesterol levels via reductions in LDLc, VLDLc, and HDLc lipoprotein fractions that were not accompanied by hepatic cholesterol accumulation. This cholesterol lowering was dependent on FXR as demonstrated in a hypothyroid-induced hypercholesterolemia setting in FXR-/- mice. In fructose-fed models, WAY-362450 also decreased TG and VLDLc levels in rats and hamsters but significantly increased HDLc levels in rats while reducing HDLc levels in hamsters. The differential effect of WAY-362450 on HDLc is likely due to a murine-specific induction of endothelial lipase and scavenger receptor-BI that does not occur in rats. These studies demonstrate a consistent ability of WAY-362450 to lower both serum TG and cholesterol levels and suggest that synthetic FXR agonists may have clinical utility in the treatment of mixed dyslipidemia. PMID- 19136378 TI - Infliximab and the TNF-alpha system. AB - Infliximab, a chimeric monoclonal antibody against TNF-alpha, is efficacious in Crohn's disease (CD) and rheumatoid arthritis (RA). Its main mechanism of action is thought to be the induction of apoptosis. The present study evaluates in detail the effects of infliximab on the TNF-alpha system using peripheral blood monocytes and T cells as well as lamina propria lymphocytes from normal individuals and patients with CD, ulcerative colitis, and RA. Lymphocytes were studied in the resting state in the absence of strong stimuli that may obscure subtle findings. Infliximab did not change the numbers of viable cells. Rather, it caused monocytes to increase their release of soluble TNFR2, which serves to neutralize TNF-alpha, potentiating the action of infliximab. It reduced TNFR2 expression, thereby decreasing TNF-alpha responsiveness. These changes were due to upregulated production of TNFR2 rather than increased shedding. Infliximab did not cause rebound production of TNF-alpha transcripts that would counteract its effects. It specifically enhanced production of IL-10 but not proinflammatory cytokines secreted by leukocytes, thereby promoting an anti-inflammatory microenvironment. In addition, infliximab caused a rise in c-Jun amino-terminal kinase phosphorylation by monocytes. Thus infliximab manipulates the TNF-alpha system to promote its anti-TNF-alpha effects. PMID- 19136379 TI - Effects of brain stem cholecystokinin-8s on gastric tone and esophageal-gastric reflex. AB - The actions of cholecystokinin (CCK) on gastrointestinal functions occur mainly via paracrine effects on peripheral sensory vagal fibers, which engage vago-vagal brain stem circuits to convey effector responses back to the gastrointestinal tract. Recent evidence suggests, however, that CCK also affects brain stem structures directly. Many electrophysiological studies, including our own, have shown that brain stem vagal circuits are excited by sulfated CCK (CCK-8s) directly, and we have further demonstrated that CCK-8s induces a remarkable degree of plasticity in GABAergic brain stem synapses. In the present study, we used fasted, anesthetized Sprague-Dawley rats to investigate the effects of brain stem administration of CCK-8s on gastric tone before and after activation of the esophageal-gastric reflex. CCK-8s microinjected in the dorsal vagal complex (DVC) or applied on the floor of the fourth ventricle induced an immediate and transient decrease in gastric tone. Upon recovery of gastric tone to baseline values, the gastric relaxation induced by esophageal distension was attenuated or even reversed. The effects of CCK-8s were antagonized by vagotomy or fourth ventricular, but not intravenous, administration of the CCK-A antagonist lorglumide, suggesting a central, not peripheral, site of action. The gastric relaxation induced by DVC microinjection of CCK-8s was unaffected by pretreatment with systemic bethanecol but was completely blocked by NG-nitro-L-arginine methyl ester, suggesting a nitrergic mechanism of action. These data suggest that 1) brain stem application of CCK-8s induces a vagally mediated gastric relaxation; 2) the CCK-8s-induced gastric relaxation is mediated via activation of nonadrenergic, noncholinergic pathways; and 3) CCK-8s reverses the esophageal gastric reflex transiently. PMID- 19136380 TI - Transcriptional regulation of IL-6 in bile duct epithelia by extracellular ATP. AB - The inflammatory cytokine IL-6 is essential for cell survival after liver injury. Bile duct epithelia (BDE) markedly upregulate IL-6 release after liver injury, but the mechanisms regulating this have not been defined. Purinergic signals induce multiple potent downstream effects in BDE, so the goals of this study were to determine whether extracellular ATP regulates BDE IL-6 transcription and to identify the molecular mechanisms regulating this process. Effects of extracellular nucleotides on IL-6 transcription in primary rat bile duct epithelia were assessed. The relative effects of cAMP and cytosolic calcium were determined by use of agonists and antagonists. The role of the cAMP response element (CRE) was determined by site-directed mutagenesis. We found that ATP potently upregulated IL-6 mRNA, and that the pharmacological profile for IL-6 upregulation was most consistent with the newly identified P2Y11 receptor. This occurred in a cAMP-dependent and calcium-dependent fashion. The effect of cAMP and calcium agonists on IL-6 promoter activity was synergistic, and mutation of the IL-6 CRE blocked upregulation by ATP. Taken together, these data show that extracellular ATP acts through a mechanism involving a rat P2Y receptor functionally related to the P2Y11 receptor, cAMP, and calcium signals and that the IL-6 promoter CRE to upregulate transcription of IL-6 in BDE. Since IL-6 has such critical importance in the liver, it is likely that this pathway is of great relevance to the understanding of hepatic response to injury. PMID- 19136381 TI - Oxidant stress-induced liver injury in vivo: role of apoptosis, oncotic necrosis, and c-Jun NH2-terminal kinase activation. AB - Oxidant stress is critically involved in various liver diseases. Superoxide formation causes c-Jun NH2-terminal kinase (JNK)- and caspase-dependent apoptosis in cultured hepatocytes. To verify these findings in vivo, male Fisher rats were treated with diquat and menadione. The oxidant stress induced by both compounds was confirmed by increased formation of glutathione disulfide and 4 hydroxynonenal protein adducts. Plasma alanine aminotransferase activities increased from 46+/-4 U/l in controls to 955+/-90 U/l at 6 h after diquat treatment. Hematoxylin and eosin staining of liver sections revealed large areas of necrotic cells at 3 and 6 h. DNA strandbreaks, evaluated with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, showed clusters of TUNEL-positive cells, where the staining was predominantly cytosolic and the cells were swollen, indicating oncotic necrosis. There was no significant increase in caspase-3 activities or relevant release of DNA fragments into the cytosol at any time between 0 and 6 h after diquat treatment. Despite the activation of JNK after high doses of diquat, the JNK inhibitor SP-600125 did not protect against diquat-induced necrosis. Menadione alone did not cause liver injury, but, in combination with phorone and FeSO4, induced moderate oncotic necrosis. On the other hand, if animals were treated with galactosamine/endotoxin as positive control for apoptosis, caspase-3 activities were increased by 259%, the number of TUNEL-positive cells with apoptotic morphology was increased 103 fold, and DNA fragmentation was enhanced 6-fold. The data indicate that liver cell death initiated by diquat-induced superoxide formation in vivo is mediated predominantly by oncotic necrosis and is independent of JNK activation. PMID- 19136382 TI - IL-4 regulates susceptibility to intestinal inflammation in murine food allergy. AB - Allergies involve a state of immediate hypersensitivity to antigens, including food proteins. The mechanism underlying the initiation and development of allergic responses involves IL-4 that directly induces the differentiation of committed effector Th2 lymphocytes. Although it is clear that Th2 responses play a pivotal role in the development of allergic responses, it remains unclear which mechanisms are involved in the development of the intestinal damages observed in food allergy. Accordingly, this work aimed to study the role of Th2/IL-4 dependent responses in the development of food allergy and intestinal pathology. C57BL/6 wild-type (WT) and IL-4-/- mice were sensitized with peanut proteins, challenged with peanut seeds, and followed for the development of food allergy and intestinal inflammation. Results demonstrated that exposure to peanut seeds led to weight loss in WT but not in IL-4-/- mice that preserved gut integrity with no signs of mucosal inflammation. These animals presented increased levels of IgG2a in sera, suggesting a role for allergic antibodies in the pathogenesis of WT animals. Most importantly, results also showed that lack of IL-4 modulated gut mucosal response in food allergy through diminished expression of TNF-alpha mRNA, increased Th1 IFN-gamma, IL-12p40, regulatory cytokines, and Foxp3, demonstrating their relevance in the control of allergic inflammatory processes, especially in the intestine. Finally, this study highlighted some of the complex mechanisms involved in the pathogenesis of allergic responses to food antigens in the gut, thereby providing valuable tools for directing novel therapeutic or preventive strategies to the control of allergic enteropathy. PMID- 19136383 TI - Reduced liver fibrosis in hypoxia-inducible factor-1alpha-deficient mice. AB - Liver fibrosis is characterized by excessive deposition of extracellular matrix in the liver during chronic injury. During early stages of this disease, cells begin to synthesize and secrete profibrotic proteins that stimulate matrix production and inhibit matrix degradation. Although it is clear that these proteins are important for development of fibrosis, what remains unknown is the mechanism by which chronic liver injury stimulates their production. In the present study, the hypothesis was tested that hypoxia-inducible factor-1alpha (HIF-1alpha) is activated in the liver during chronic injury and regulates expression of profibrotic proteins. To investigate this hypothesis, mice were subjected to bile duct ligation (BDL), an animal model of liver fibrosis. HIF 1alpha protein was increased in the livers of mice subjected to BDL by 3 days after surgery. To test the hypothesis that HIF-1alpha is required for the development of fibrosis, control and HIF-1alpha-deficient mice were subjected to BDL. Levels of type I collagen and alpha-smooth muscle actin mRNA and protein were increased in control mice by 14 days after BDL. These levels were significantly reduced in HIF-1alpha-deficient mice. Next, the levels of several profibrotic mediators were measured to elucidate the mechanism by which HIF 1alpha promotes liver fibrosis. Platelet-derived growth factor (PDGF)-A, PDGF-B, and plasminogen activator inhibitor-1 mRNA levels were increased to a greater extent in control mice subjected to BDL compared with HIF-1alpha-deficient mice at 7 and 14 days after BDL. Results from these studies suggest that HIF-1alpha is a critical regulator of profibrotic mediator production during the development of liver fibrosis. PMID- 19136384 TI - Increased susceptibility to exacerbated liver injury in hypercholesterolemic ApoE deficient mice: potential involvement of oxysterols. AB - The contribution of metabolic factors to the severity of liver disease is not completely understood. In this study, apolipoprotein E-deficient (ApoE-/-) mice were evaluated to define potential effects of hypercholesterolemia on the severity of carbon tetrachloride (CCl4)-induced liver injury. Under baseline conditions, hypercholesterolemic ApoE-/- mice showed increased hepatic oxidative stress (SOD activity/4-hydroxy-2-nonenal immunostaining) and higher hepatic TGF beta1, MCP-1, and TIMP-1 expression than wild-type control mice. After CCl4 challenge, ApoE-/- mice exhibited exacerbated steatosis (Oil Red O staining), necroinflammation (hematoxylin-eosin staining), macrophage infiltration (F4/80 immunohistochemistry), and fibrosis (Sirius red staining and alpha-smooth muscle actin immunohistochemistry) and more severe liver injury [alanine aminotransferase (ALT) and aspartate aminotransferase] than wild-type controls. Direct correlations were identified between serum cholesterol and hepatic steatosis, fibrosis, and ALT levels. These changes did not reflect the usual progression of the disease in ApoE-/- mice, since exacerbated liver injury was not present in untreated age-paired ApoE-/- mice. Moreover, hepatic cytochrome P 450 expression was unchanged in ApoE-/- mice. To explore potential mechanisms, cell types relevant to liver pathophysiology were exposed to selected cholesterol oxidized products. Incubation of hepatocytes with a mixture of oxysterols representative of those detected by GC-MS in livers from ApoE-/- mice resulted in a concentration-dependent increase in total lipoperoxides and SOD activity. In hepatic stellate cells, oxysterols increased IL-8 secretion through a NF-kappaB independent mechanism and upregulated TIMP-1 expression. In macrophages, oxysterols increased TGF-beta1 secretion and MCP-1 expression in a concentration dependent manner. Oxysterols did not compromise cell viability. Taken together, these findings demonstrate that hypercholesterolemic mice are sensitized to liver injury and that cholesterol-derived products (i.e., oxysterols) are able to induce proinflammatory and profibrogenic mechanisms in liver cells. PMID- 19136386 TI - Curriculum evaluation and revision in a nascent field: the utility of the retrospective pretest--posttest model in a homeland security program of study. AB - Although most academic disciplines evolve at a measured pace, the emerging field of homeland security must, for reasons of safety and security, evolve rapidly. The Department of Homeland Security sponsored the establishment of a graduate educational program for key officials holding homeland security roles. Because homeland security is a nascent field, the establishment of a program curriculum was forced to draw from a variety of disciplines. Curriculum evaluation was complicated by the rapid changes occurring in the emerging discipline, producing response shift bias, and interfering with the pre-post assessments. To compensate for the validity threat associated with response shift bias, a retrospective pretest-posttest evaluative methodology was used. Data indicate the program has evolved in a significant and orderly fashion and these data support the use of this innovative evaluation approach in the development of any discipline. PMID- 19136385 TI - Lipopolysaccharide activates innate immune responses in murine intestinal myofibroblasts through multiple signaling pathways. AB - Myofibroblasts (MF) play an important role in intestinal wound healing. A compromised epithelial barrier exposes intestinal subepithelial MF to luminal bacterial products. However, responses of murine intestinal MF to bacterial adjuvants and potential roles of intestinal MF in innate immune responses are not well defined. Our aims in this study were to determine innate immune responses and intracellular signaling pathways of intestinal MF exposed to LPS, a prototypic Toll-like receptor (TLR) ligand. Expression of TLR4 in primary murine intestinal MF cultures was confirmed by RT-PCR and Western blotting. LPS-induced secretion of prostaglandin E2 (PGE2), interleukin (IL)-6, and keratinocyte derived chemokines (KC) was measured by ELISA. Intracellular responses to LPS were assessed by Western blotting for NF-kappaB p65, Ikappa-Balpha, Akt, p38 MAP kinase, and cyclooxygenase-2 (COX-2). LPS induced rapid phosphorylation of NF kappaB p65, Akt, and p38 MAPK and degradation of Ikappa-Balpha. LPS induced expression of COX-2 and secretion of PGE2 (2.0+/-0.8-fold induction vs. unstimulated cells), IL-6 (6.6+/-0.4-fold induction), and KC (12.5+/-0.4-fold induction). Inhibition of phosphoinositide-3 (PI3)-kinase, p38 MAPK, or NF-kappaB pathways reduced LPS-induced PGE2, IL-6, and KC secretion. These studies show that primary murine intestinal MF respond to LPS, evidenced by activation of NF kappaB, PI3-kinase, and MAPK signaling pathways and secretion of proinflammatory molecules. Inhibition of these pathways attenuated LPS-dependent PGE2, IL-6, and KC production, indicating that LPS activates MF by multiple signaling pathways. These data support the hypothesis that MF are a component of the innate immune system and may exert paracrine effects on adjacent epithelial and immune cells by responding to luminal bacterial adjuvants. PMID- 19136388 TI - Androgen receptor in Sertoli cells is not required for testosterone-induced suppression of spermatogenesis, but contributes to Sertoli cell organization in Utp14b jsd mice. AB - Testosterone acting through the androgen receptor (AR) maintains the arrest of spermatogonial differentiation in juvenile spermatogonial depletion (jsd mutation in the Utp14b gene) mutant adult male mice. It is not known which of the somatic cell types expressing AR mediates this inhibition. To determine whether Sertoli cells are responsible, we selectively eliminated AR in Sertoli cells in jsd mice containing a floxed-Ar gene and an anti-Mullerian hormone-Cre transgene. In these Sertoli AR-knockout (SCARKO)-jsd mice, spermatogonial differentiation did not recover. However, the normal organization of Sertoli cell nuclei was drastically disrupted in SCARKO-jsd mice compared with SCARKO or jsd mice. In addition, the extent of ectoplasmic specializations was reduced; tight junctions were not found; vinculin, an anchoring protein found in ectoplasmic specializations, became uniformly distributed in the cytoplasm; and the adult Sertoli cells showed excess heterochromatin subjacent to their nuclear envelope. Despite the abnormalities in Sertoli cells in SCARKO-jsd mice, global suppression of testosterone action and levels was still effective in restoring the differentiated germ cells, and this was accompanied by an improved arrangement of Sertoli cell nuclei. We conclude that Sertoli cells are not targets for the testosterone-mediated inhibition of spermatogonial differentiation in jsd mice, and that both AR in Sertoli cells and the presence of differentiated germ cells contribute to maintaining the organization of Sertoli cells within the seminiferous tubules. PMID- 19136389 TI - Immunolocalization of androgen receptors, estrogen alpha receptors, and estrogen beta receptors in experimentally induced canine prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is an age-dependent prostatic disease affecting male humans and dogs. In dogs, the combined administration of estrogens and androgens synergistically increases prostate weight, and continued treatment leads to the development of glandular hyperplasia. The aim of the present study was to examine the immunohistochemical expression of androgen receptor (AR), estrogen receptor alpha (ER alpha), and estrogen receptor beta (ER beta) in the different cell types of the prostate gland in an experimental model. Five male beagle dogs were castrated and treated with 25 mg of 5 alpha-androstane-3 alpha and 17beta-diol and 0.25 mg 17beta-estradiol for 30 weeks. Prostate specimens were surgically obtained every 45 days (experimental stages M0 to M6: 0, 12, 18, 24, 30, and 36 weeks from the beginning of the hormonal treatment). The control group consisted of 3 noncastrated dogs treated with a vehicle, from which specimens were only taken at the time points M0, M1, M4, and M6. Immunohistochemical data revealed high AR and ER alpha expression in the epithelial and stromal cell nuclei of all the experimental and control specimens. Weak staining of the cytoplasm was observed only in epithelial cells. The suspension of hormone treatment led to a significant reduction in the expression of both receptors. On the contrary, ER beta was expressed only in epithelial cell nuclei, with no significant differences in the percentages of stained nuclei between control and hormonally treated or atrophic prostates. Results indicate that AR, ER alpha, and ER beta are differently expressed in canine prostate tissue and that they show specific expression patterns in response to the hormonal induction of BPH. PMID- 19136391 TI - Increased aggressiveness of human prostate PC-3 tumor cells expressing cell surface localized membrane type-1 matrix metalloproteinase (MT1-MMP). AB - Membrane type-1 matrix metalloproteinase (MT1-MMP) is a multidomain transmembrane endopeptidase with a major role in physiological and pathological processes through proteolysis of extracellular matrix and other pericellular proteins. We examined cell surface function of MT1-MMP in PC-3 human prostate tumor cells selected for metastasis in nude mice (PC-3-LN4), or transfected with the full length wild-type (WT) MT1-MMP or with the mutant form lacking the cytoplasmic tail (Delta C-MT1-MMP). Enhanced cell surface MT1-MMP was determined by fluorescence-activated cell sorting analysis and evidenced mechanistically by increased activation of proMMP-2 and invasion into type-I collagen gels. PC-3 cells overexpressing MT1-MMP grew faster than mock-transfected control cells subcutaneously in nude mice. MT1-MMP localized in caveolae, as judged by immunofluorescence microscopy and sucrose-gradient, detergent-resistant cell fractionation. Delta C-MT1-MMP was strongly associated with caveolae, whereas the WT form was present in both caveolae and noncaveolae fractions. The role of plasma membrane MT1-MMP was supported by localization of MT1-MMP by immunofluorescence microscopy at the cell surface of tumor cells in primary prostate cancers. Increased plasma membrane localization of MT1-MMP, either in caveolae or in other lipid raft structures, is a mechanism to localize this proteinase in contact with extracellular matrix and other pericellular proteins, the cleavage of which can facilitate prostate cancer cell invasion and metastasis. PMID- 19136390 TI - Estrogen enhances recovery from radiation-induced spermatogonial arrest in rat testes. AB - Irradiation of LBNF(1) rat testes induces spermatogonial differentiation arrest, which can be reversed by gonadotropin-releasing hormone (GnRH) antagonist-induced suppression of intratesticular testosterone (ITT) and follicle-stimulating hormone (FSH). Although exogenous estrogen treatment also enhanced spermatogenic recovery, as measured by the tubule differentiation index (TDI), it was not clear whether estrogen stimulated spermatogonial differentiation only by further suppressing ITT or by an additional independent mechanism as well. To resolve this question, we performed the following experiments. At 15 weeks after irradiation, rats were treated with GnRH antagonist; some also received 17beta estradiol (E2) and were killed 4 weeks later. GnRH antagonist treatment increased the TDI from 0% to 8%, and addition of E2 further increased the TDI to 39%. However, E2 addition further reduced ITT from 7 ng/g testis, observed with GnRH antagonist to 3 ng/g testis, so decreased ITT levels might have contributed to recovery. Next GnRH antagonist-treated rats were given exogenous testosterone and flutamide to stabilize ITT levels and block its action. This increased TDI slightly from 8% to 13%, but the further addition of E2 significantly raised the TDI to 27%, indicating it acted by a mechanism independent of ITT levels. Plots of TDI for all treatment groups compared with ITT, FSH, or a linear combination of ITT and FSH showed that treatments including E2 produced higher TDI values than did treatments without E2. These results indicate that there was an effect of E2 on spermatogonial differentiation because of an additional direct action on the testis that is unrelated to its suppression of testosterone or gonadotropins. PMID- 19136392 TI - Effects of an environmentally relevant organochlorine mixture and a metabolized extract of this mixture on porcine sperm parameters in vitro. AB - Organochlorine chemicals are present in the environment worldwide; however, populations living in the Far North are particularly at risk because their traditional diets are mainly composed of contaminated animals (fish, seals, whales, and polar bears). It has been suggested that male fertility is globally declining, possibly because of chronic, low-level exposure to environmental contaminants. This study was designed to assess the effects on fresh sperm fertility parameters using the porcine model of 1) an environmentally relevant mixture of 15 organochlorines and 2) the metabolized extract of this mixture. In the first experiment, the organochlorine mixture (at relative concentrations of 10.5, 14.7, and 21 microg/mL polychlorinated biphenyls [PCBs]) reduced sperm total motility, progressive motility, and viability, and increased capacitation, spontaneous acrosome reaction rates, and cytosolic calcium levels, suggesting that the mixture alters the sperm membrane and is detrimental to sperm function. In the second experiment, the metabolized extract of this organochlorine mixture (at relative concentrations of 0.9, 1.8, 2.7, 3.6, and 4.5 microg/L OH-PCBs) tended to decrease only sperm total motility. In an in vitro porcine model, the mixture of organochlorines, as found in the Arctic food chain, was rapidly detrimental to sperm function at concentrations above environmental levels. In contrast, short and physiologically relevant exposure to the metabolized extract of this mixture induced only limited adverse effects on sperm motility. PMID- 19136394 TI - Breastfeeding mothers returning to work: possibilities for information, anticipatory guidance and support from US health care professionals. AB - Today, more mothers in the United States are in the labor force, and returning to the workforce presents numerous challenges for the breastfeeding mother. Although it as been demonstrated that maternal employment is associated with a decrease in the length of time a mother continues to breastfeed, health care providers are in a unique position to enhance a mother's breastfeeding success as she transitions back into the workplace. This article describes various commonly perceived obstacles to combining breastfeeding and working and provides examples of information, anticipatory guidance, and support that health care providers can use to assist a breastfeeding mother with a successful return to the workforce. PMID- 19136393 TI - Testicular function in poor-risk nonseminomatous germ cell tumors treated with methotrexate, paclitaxel, ifosfamide, and cisplatin combination chemotherapy. AB - Our objective was to investigate the impact of methotrexate, paclitaxel, ifosfamide, and cisplatin (M-TIP) on long-term fertility in poor-risk nonseminomatous germ cell tumors (NSGCT). Thirty patients with poor-risk NSGCT (median age, 29 years; range, 17-62 years) were treated with methotrexate 250 mg/m(2) with folinic acid rescue (day 1) and paclitaxel 175 mg/m(2) (day 1), followed by ifosfamide 1.2 g/m(2) and cisplatin 20 mg/m(2) (days 2-6). Treatment consisted of 4 cycles of M-TIP administered every 3 weeks. Twenty-one patients were continuously disease-free at a median follow-up of 5.3 years (range, 0.9-8.4 years). Sperm count and hormonal analyses were examined prechemotherapy (30 patients) and postchemotherapy (21 patients). Counts were classified as follows: lower than 1 x 10(6)/mL, azoospermia; 1-20 x 10(6)/mL, oligospermia (OS); higher than 20 x 10(6)/mL, normospermia (NS). Patients were followed for a median of 2.3 years (range, 0.9-3.8 years) postchemotherapy. The prechemotherapy median luteinizing hormone (LH) serum levels were slightly above the upper normal limit, whereas the serum levels of follicle-stimulating hormone (FSH) and testosterone (T) were within the reference interval. Eleven (52.3%) patients had NS prechemotherapy. Among the patients with NS, 72.7% still had NS following chemotherapy. Overall, 17 of 21 (80.9%; 33.3% OS and 47.6% NS) patients had recovery of spermatogenesis after treatment. The median FSH serum levels were significantly elevated at least 1 year postchemotherapy when compared with the pretreatment levels. Eighteen months after the completion of chemotherapy the median FSH levels had returned to the reference limits. Serum LH and T levels were unaffected by chemotherapy. Prior to chemotherapy 4 of 30 patients had fathered 5 children. Since completion of chemotherapy, 5 patients have fathered 5 children. The majority of men with poor-risk germ cell tumors who were treated with the M-TIP regimen demonstrated recovery spermatogenesis after treatment, and Leydig cell function was unaffected. PMID- 19136395 TI - Transfer of buprenorphine into breast milk and calculation of infant drug dose. AB - Little is known about the safety of buprenorphine (BUP) in breastfeeding. The aim of this work was to investigate the transfer of buprenorphine and its main active metabolite, norbuprenorphine (n-BUP), into human milk and to determine the drug dose and effects in exposed infants. Seven lactating women, who were maintained on BUP treatment because of previous opiate addiction, were studied in an open observational study. All mothers had a strong wish to breastfeed their newborn infants. Buprenorphine samples for analysis were collected from the urine of 6 infants together with breast milk, blood, and urine from their mothers during a 24-hour period in the week after birth. One mother-infant pair was studied at 9 months of age. Buprenorphine and n-BUP were analyzed by a liquid chromatography/mass spectrometry method suitable for handling different matrices. Buprenorphine and n-BUP were found in low levels in the infants' urine. Breastfed infants were exposed to a calculated BUP dose per kg bodyweight less than 1%, with an average milk/plasma area under the curve of 1.7 (range, 1.1-2.8) for BUP and 0.7 (range, 0.4-1.2) for n-BUP. These data support the use of BUP during breastfeeding. However, the authors recommend that infants be monitored closely. PMID- 19136396 TI - Sources and acceptance of infant-feeding advice among low-income women. AB - The purpose of this study is to identify sources and acceptability of infant feeding advice among participants in the US-based Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). Focus groups are used to identify sources of infant-feeding advice and factors that contribute to acceptance of or resistance to that advice among 65 WICeligible mothers (34 English speaking and 31 Spanish speaking). The mothers primarily rely on experienced family and friends for advice and frequently use their own intuition to find solutions that work to solve real or perceived infant-feeding problems. Professional advice is perceived as credible when caregivers exhibit characteristics similar to those of experienced family and friends: confidence, empathy, respect, and calm. Using this information, it may be possible for WIC staff to make programmatic modifications to increase their ability to promote optimal infant-feeding behaviors in this population, thereby contributing to the reduction in the prevalence of childhood overweight. PMID- 19136397 TI - Caring for AIDS patients in the ICU: expanding horizons. PMID- 19136398 TI - Primum non nocere: how to cause chaos with a bronchoscope in the ICU. PMID- 19136399 TI - The old and the new of sedation for bronchoscopy. PMID- 19136400 TI - Interactive physiology grand rounds: introduction to the series. PMID- 19136401 TI - Pectoriloquy, a retrospective analysis. PMID- 19136403 TI - Implementation of ICU palliative care guidelines and procedures: a quality improvement initiative following an investigation of alleged euthanasia. AB - Ethical conflicts are commonly encountered in the course of delivering end-of life care in the ICU. Some ethical concerns have legal dimensions, including concerns about inappropriate hastening of death. Despite these concerns, many ICUs do not have explicit policies and procedures for withdrawal of life sustaining treatments. We describe a US Office of Inspector General (OIG) investigation of end-of-life care practices in our ICU. The investigation focused on care delivered to four critically ill patients with terminal diseases and an ICU nurse's concern that the patients had been subjected to euthanasia. The OIG investigation also assessed the validity of allegations that patient flow in and out of our ICU was inappropriately influenced by scheduled surgeries and that end of-life care policies in our ICU were not clear. Although the investigation did not substantiate the allegations of euthanasia or inappropriate ICU patient flow, it did find that the policies that discuss end-of-life care issues were not clear and allowed for wide-ranging interpretations. Acting on the OIG recommendations, we developed a quality improvement initiative addressing end-of-life care in our ICU, intended to enhance communication and understanding about palliative care practices in our ICU, to prevent ethical conflicts surrounding end-of-life care, and to improve patient care. The initiative included the introduction of newly developed ICU comfort care guidelines, a physician order set, and a physician template note. Additionally, we implemented an educational program for ICU staff. Staff feedback regarding the initiative has been highly favorable, and the nurse whose concerns led to the investigation was satisfied not only with the investigation but also the policies and procedures that were subsequently introduced in our ICU. PMID- 19136404 TI - Vicks VapoRub induces mucin secretion, decreases ciliary beat frequency, and increases tracheal mucus transport in the ferret trachea. AB - BACKGROUND: Vicks VapoRub (VVR) [Proctor and Gamble; Cincinnati, OH] is often used to relieve symptoms of chest congestion. We cared for a toddler in whom severe respiratory distress developed after VVR was applied directly under her nose. We hypothesized that VVR induced inflammation and adversely affected mucociliary function, and tested this hypothesis in an animal model of airway inflammation. METHODS: [1] Trachea specimens excised from 15 healthy ferrets were incubated in culture plates lined with 200 mg of VVR, and the mucin secretion was compared to those from controls without VVR. Tracheal mucociliary transport velocity (MCTV) was measured by timing the movement of 4 microL of mucus across the trachea. Ciliary beat frequency (CBF) was measured using video microscopy. [2] Anesthetized and intubated ferrets inhaled a placebo or VVR that was placed at the proximal end of the endotracheal tube. We evaluated both healthy ferrets and animals in which we first induced tracheal inflammation with bacterial endotoxin (a lipopolysaccharide [LPS]). Mucin secretion was measured using an enzyme-linked lectin assay, and lung water was measured by wet/dry weight ratios. RESULTS: [1] Mucin secretion was increased by 63% over the controls in the VVR in vitro group (p < 0.01). CBF was decreased by 35% (p < 0.05) in the VVR group. [2] Neither LPS nor VVR increased lung water, but LPS decreased MCTV in both normal airways (31%) and VVR-exposed airways (30%; p = 0.03), and VVR increased MCTV by 34% in LPS-inflamed airways (p = 0.002). CONCLUSIONS: VVR stimulates mucin secretion and MCTV in the LPS-inflamed ferret airway. This set of findings is similar to the acute inflammatory stimulation observed with exposure to irritants, and may lead to mucus obstruction of small airways and increased nasal resistance. PMID- 19136405 TI - COPD as a disease of accelerated lung aging. AB - There is increasing evidence for a close relationship between aging and chronic inflammatory diseases. COPD is a chronic inflammatory disease of the lungs, which progresses very slowly and the majority of patients are therefore elderly. We here review the evidence that accelerating aging of lung in response to oxidative stress is involved in the pathogenesis and progression of COPD, particularly emphysema. Aging is defined as the progressive decline of homeostasis that occurs after the reproductive phase of life is complete, leading to an increasing risk of disease or death. This results from a failure of organs to repair DNA damage by oxidative stress (nonprogrammed aging) and from telomere shortening as a result of repeated cell division (programmed aging). During aging, pulmonary function progressively deteriorates and pulmonary inflammation increases, accompanied by structural changes, which are described as senile emphysema. Environmental gases, such as cigarette smoke or other pollutants, may accelerate the aging of lung or worsen aging-related events in lung by defective resolution of inflammation, for example, by reducing antiaging molecules, such as histone deacetylases and sirtuins, and this consequently induces accelerated progression of COPD. Recent studies of the signal transduction mechanisms, such as protein acetylation pathways involved in aging, have identified novel antiaging molecules that may provide a new therapeutic approach to COPD. PMID- 19136406 TI - Critical illness-related corticosteroid insufficiency. AB - The diagnosis of adrenal failure and the indications for corticosteroid therapy in critically ill patients are controversial. This controversy is fueled by the complexity of the issues and the paucity of data from high quality clinical trials. Nevertheless, while the use of high-dose corticosteroids in patients with severe sepsis and ARDS failed to improve outcome and was associated with increased complications, an extended course of stress-dose corticosteroids has been reported to increase the occurrence of ventilator-free days and survival in select groups of ICU patients. These patients typically have an exaggerated proinflammatory response. Until recently the exaggerated proinflammatory response that characterizes critically ill patients with systemic inflammation has focused on suppression of the hypothalamic-pituitary-adrenal axis and adrenal failure. However, experimental and clinical data suggest that glucocorticoid tissue resistance may also play an important role. This complex syndrome is referred to as critical illness-related corticosteroid insufficiency (CIRCI) and is defined as inadequate corticosteroid activity for the severity of the illness of a patient. The paper reviews cortisol physiology, CIRCI, and the role of corticosteroid therapy in critically ill patients. PMID- 19136407 TI - Assessment of pleural pressure in the evaluation of pleural effusions. PMID- 19136408 TI - Exercise-induced bronchospasm: coding and billing for physician services. AB - Physician reporting of the service to insurance companies for reimbursement is multifaceted and perplexing to those who do not understand the factors to consider. Test selection should be individualized based on the patient's history and/or needs. Federal regulations concerning physician supervision of diagnostic tests mandate different levels of physician supervision based on the type and complexity of the test. Many factors play a key role in physician claim submission. These include testing location, component services, coding edits, and additional visits. Medical necessity of the service(s) must also be demonstrated for payer consideration and reimbursement. The following article reviews various tests for exercise-induced bronchospasm and focuses on issues to assist the physician in reporting the services accurately and appropriately. PMID- 19136409 TI - Shock complicating successful bronchial artery embolization for severe hemoptysis. AB - Bronchial artery embolization (BAE) complications are uncommon. We describe three patients who had a shock related to splenic infarction after a successful BAE. PMID- 19136410 TI - A 79-year-old man with chest wall pain and a rapidly growing mass. PMID- 19136411 TI - A 22-year-old man with headache and stiff neck after a water skiing fall. PMID- 19136412 TI - A hypotensive 22-year-old man with chest pain, cough, and an abnormal chest radiograph. PMID- 19136413 TI - When does previous disclosure become a 'prior publication' problem? PMID- 19136414 TI - Dyspnea in a 43-year-old woman with polycystic kidney disease. PMID- 19136417 TI - The need for expertise and the scientific base of evidence-based medicine. PMID- 19136418 TI - Continuous critical care and long-term noninvasive ventilatory support for patients with neuromuscular disease. PMID- 19136419 TI - Dysglycemia in critically ill patients from medicine. PMID- 19136420 TI - Smoking cessation counseling in Orthodox Jewish populations. PMID- 19136421 TI - Neuromuscular control of trunk stability: clinical implications for sports injury prevention. PMID- 19136422 TI - Surgical treatment for chronic disease and disorders of the achilles tendon. AB - Chronic Achilles tendon disorders range from overuse syndromes to frank ruptures. Numerous forms of treatment have been used, depending on the nature of the disorder or injury. Ultrasonography and magnetic resonance imaging are commonly used for evaluation. The spectrum of disease comprises paratenonitis, tendinosis, paratenonitis with tendinosis, retrocalcaneal bursitis, insertional tendinosis, and chronic rupture. However, there is no clear consensus on what defines a chronic Achilles disorder. Nonsurgical therapy is the mainstay of treatment for most patients with overuse syndromes. Surgical techniques for overuse syndromes or chronic rupture include debridement, local tissue transfer, augmentation, and synthetic grafts. Local tissue transfer most commonly employs either the flexor hallucis longus or flexor digitorum longus tendon to treat a chronic rupture. Reports on long-term outcomes are needed before useful generalizations can be made regarding treatment. PMID- 19136423 TI - Traumatic hip dislocations in children and adolescents: pitfalls and complications. AB - Traumatic hip dislocation is an uncommon injury in children. Lack of familiarity with management of the treating physician may lead to complications. Hip dislocation in young children can occur with minor trauma; in adolescents, greater force is required to produce a traumatic complete hip dislocation. Transient hip dislocation with spontaneous but incomplete reduction is a diagnostic pitfall that can occur in adolescents. Any asymmetric widening of the hip joint warrants additional investigation. Most dislocations in children can be reduced with gentle manipulation. Urgent reduction of the hip within 6 hours of injury reduces the risk of osteonecrosis. However, closed reduction in adolescents should be performed with caution because of the risk of displacement of the femoral head during manipulation. Open reduction is indicated when closed reduction fails or when there is interposition of bone or soft tissue following attempted closed reduction. Late complications include osteonecrosis, coxa magna, and osteoarthritis. PMID- 19136424 TI - Magnetic resonance imaging of the spine. AB - Historically, magnetic resonance imaging has offered poor specificity in the diagnosis of back pain. Researchers currently are engaged in developing new techniques, and clinicians are successfully utilizing existing technologies (ie, diffusion-weighted imaging) that previously were not used to evaluate the spine. Magnetic resonance imaging may be used in several spinal applications: intervertebral disk and facet joint degeneration, spinal canal stenosis, suspected diskitis or osteomyelitis, suspected spinal column neoplasia, vascular disorders, trauma, and demyelinating disease. PMID- 19136425 TI - Knee injury outcomes measures. AB - Outcomes measures have long been used in the assessment of knee injuries and management protocols. In the past decade, there has been a shift from clinician based outcomes tools to the development and validation of patient-reported outcomes measures. General health as well as disease- and medical condition specific outcomes measures have been so modified. The Medical Outcomes Study 36 Item Short Form is the most commonly used general health measure in orthopaedics. Joint-specific measures include the Western Ontario and McMaster Universities Osteoarthritis Index, the Knee Injury and Osteoarthritis Outcome Score, and the International Knee Documentation Committee Subjective Form. The Lysholm Knee Scale and the Cincinnati Knee Rating Scale continue to be popular, especially for the assessment of ligamentous injuries. The ACL Quality of Life score is a disease-specific, patient-reported outcomes measure of anterior cruciate ligament deficiency. The historically used Tegner activity level scale and the recently developed Marx activity level scale are used in conjunction with these outcomes measures to make possible a global assessment of recovery from knee injuries and clinician interventions. PMID- 19136426 TI - Soft-tissue sarcoma. AB - Soft-tissue sarcomas are rare malignancies of mesodermal origin. Common sites of involvement include the extremities, trunk, retroperitoneum, and the head and neck. Soft-tissue sarcomas of the extremities and pelvis are the most relevant to the orthopaedic surgeon. The patient with extremity soft-tissue sarcoma typically presents with a painless, enlarging mass. Advanced imaging techniques and biopsy, which are best done at tertiary referral centers, usually confirm the diagnosis. Factors such as sarcoma size, location, grade, histologic subtype, and stage, as well as patient age and comorbidities, determine the specific approach to management and patient outcome. Limb-sparing surgical resection is the mainstay of treatment. Radiation is used for unresectable tumors and as a neoadjuvant or an adjuvant to resection. The use of chemotherapy is controversial, and no standardized protocol has been established. PMID- 19136427 TI - Parkinson's disease and the orthopaedic patient. AB - Parkinson's disease is a progressive neurologic disorder that affects the musculoskeletal system in multiple ways. As medication and surgical management of this disorder have improved, the life spans and quality of life of patients affected by it also have improved. With age, the risk of fracture, osteoarthritis, and osteopenia increase in patients with Parkinson's disease compared with the general population. The symptoms of Parkinson's disease predispose patients to gait abnormalities and loss of bone mass, which commonly result in falls and fracture. Although preventive measures such as medication, lifestyle changes, and vitamin replacement may help, surgical intervention is often indicated. Surgical treatment and postoperative management of both elective and emergent surgery are complicated and controversial. PMID- 19136429 TI - Hepatic lipase maturation: a partial proteome of interacting factors. AB - Tandem affinity purification (TAP) has been used to isolate proteins that interact with human hepatic lipase (HL) during its maturation in Chinese hamster ovary cells. Using mass spectrometry and Western blotting, we identified 28 proteins in HL-TAP isolated complexes, 16 of which localized to the endoplasmic reticulum (ER), the site of HL folding and assembly. Of the 12 remaining proteins located outside the ER, five function in protein translation or ER-associated degradation (ERAD). Components of the two major ER chaperone systems were identified, the BiP/Grp94 and the calnexin (CNX)/calreticulin (CRT) systems. All factors involved in CNX/CRT chaperone cycling were identified, including UDP glucose:glycoprotein glucosyltransferase 1 (UGGT), glucosidase II, and the 57 kDa oxidoreductase (ERp57). We also show that CNX, and not CRT, is the lectin chaperone of choice during HL maturation. Along with the 78 kDa glucose-regulated protein (Grp78; BiP) and the 94 kDa glucose-regulated protein (Grp94), an associated peptidyl-prolyl cis-trans isomerase and protein disulfide isomerase were also detected. Finally, several factors in ERAD were identified, and we provide evidence that terminally misfolded HL is degraded by the ubiquitin mediated proteasomal pathway. We propose that newly synthesized HL emerging from the translocon first associates with CNX, ERp57, and glucosidase II, followed by repeated posttranslational cycles of CNX binding that is mediated by UGGT. BiP/Grp94 may stabilize misfolded HL during its transition between cycles of CNX binding and may help direct its eventual degradation. PMID- 19136432 TI - Current issues and considerations regarding trichomoniasis and human immunodeficiency virus in African-Americans. AB - Trichomonas vaginalis has long been recognized as one of the most prevalent sexually transmitted infections. However, it is only in recent years that it has been appreciated that Trichomonas may play a critical role in amplifying human immunodeficiency virus (HIV) transmission. Given the evidence that T. vaginalis likely promotes HIV infection, the apparent high level of Trichomonas infection in the African-American community is cause for concern. Even if T. vaginalis increases the risk of HIV transmission by a small or modest amount, it translates into a sizable population effect since Trichomonas is so common in this community. Therefore, control of trichomoniasis may represent an important avenue of control for the prevention of HIV transmission, particularly among African Americans. PMID- 19136431 TI - Acquired immunity to malaria. AB - Naturally acquired immunity to falciparum malaria protects millions of people routinely exposed to Plasmodium falciparum infection from severe disease and death. There is no clear concept about how this protection works. There is no general agreement about the rate of onset of acquired immunity or what constitutes the key determinants of protection; much less is there a consensus regarding the mechanism(s) of protection. This review summarizes what is understood about naturally acquired and experimentally induced immunity against malaria with the help of evolving insights provided by biotechnology and places these insights in the context of historical, clinical, and epidemiological observations. We advocate that naturally acquired immunity should be appreciated as being virtually 100% effective against severe disease and death among heavily exposed adults. Even the immunity that occurs in exposed infants may exceed 90% effectiveness. The induction of an adult-like immune status among high-risk infants in sub-Saharan Africa would greatly diminish disease and death caused by P. falciparum. The mechanism of naturally acquired immunity that occurs among adults living in areas of hyper- to holoendemicity should be understood with a view toward duplicating such protection in infants and young children in areas of endemicity. PMID- 19136433 TI - Microbiology of odontogenic bacteremia: beyond endocarditis. AB - SUMMARY: The human gingival niche is a unique microbial habitat. In this habitat, biofilm organisms exist in harmony, attached to either enamel or cemental surfaces of the tooth as well as to the crevicular epithelium, subjacent to a rich vascular plexus underneath. Due to this extraordinary anatomical juxtaposition, plaque biofilm bacteria have a ready portal of ingress into the systemic circulation in both health and disease. Yet the frequency, magnitude, and etiology of bacteremias due to oral origin and the consequent end organ infections are not clear and have not recently been evaluated. In this comprehensive review, we address the available literature on triggering events, incidence, and diversity of odontogenic bacteremias. The nature of the infective agents and end organ infections (other than endocarditis) is also described, with an emphasis on the challenge of establishing the link between odontogenic infections and related systemic, focal infections. PMID- 19136434 TI - Host-microbe interactions and defense mechanisms in the development of amoebic liver abscesses. AB - SUMMARY: Amoebiasis by Entamoeba histolytica is a major public health problem in developing countries and leads to several thousand deaths per year. The parasite invades the intestine (provoking diarrhea and dysentery) and the liver, where it forms abscesses (amoebic liver abscesses [ALAs]). The liver is the organ responsible for filtering blood coming from the intestinal tract, a task that implies a particular structure and immune features. Amoebae use the portal route and break through the sinusoidal endothelial barrier to reach the hepatic parenchyma. When faced with systemic and cell-mediated defenses, trophozoites adapt to their new environment and modulate host responses, leading to parasite survival and the formation of inflammatory foci. Cytopathogenic effects and the onset of inflammation may be caused by diffusible products originating from parasites and/or immune cells either by their secretion or by their release after cell death. Liver infection thus results from the interplay between E. histolytica and hepatic cells. Despite its importance in terms of public health burden, the lack of integrated data on ALA genesis means that we have only an incomplete description of the initiation and development of hepatic amoebiasis. Here, we review the main steps of ALA development as well as the responses triggered in both the host and the parasite. Transcriptome studies highlighted parasite factors involved in adherence to human cells, cytopathogenic effects, and adaptative and stress responses. An understanding of their role in ALA development will help to unravel the host-pathogen interactions and their evolution throughout the infection. PMID- 19136435 TI - Immunobiology of human cytomegalovirus: from bench to bedside. AB - SUMMARY: Following primary infection, human cytomegalovirus (HCMV) establishes lifelong latency and periodically reactivates without causing symptoms in healthy individuals. In the absence of an adequate host-derived immune response, this fine balance of permitting viral reactivation without causing pathogenesis is disrupted, and HCMV can subsequently cause invasive disease and an array of damaging indirect immunological effects. Over the last decade, our knowledge of the immune response to HCMV infection in healthy virus carriers and diseased individuals has allowed us to translate these findings to develop better diagnostic tools and therapeutic strategies. The application of these emerging technologies in the clinical setting is likely to provide opportunities for better management of patients with HCMV-associated diseases. PMID- 19136437 TI - Epidemiology, diagnosis, treatment, and control of trichinellosis. AB - SUMMARY: Throughout much of the world, Trichinella spp. are found to be the causative agents of human trichinellosis, a disease that not only is a public health hazard by affecting human patients but also represents an economic problem in porcine animal production and food safety. Due to the predominantly zoonotic importance of infection, the main efforts in many countries have focused on the control of Trichinella or the elimination of Trichinella from the food chain. The most important source of human infection worldwide is the domestic pig, but, e.g., in Europe, meats of horses and wild boars have played a significant role during outbreaks within the past 3 decades. Infection of humans occurs with the ingestion of Trichinella larvae that are encysted in muscle tissue of domestic or wild animal meat. Early clinical diagnosis of trichinellosis is rather difficult because pathognomonic signs or symptoms are lacking. Subsequent chronic forms of the disease are not easy to diagnose, irrespective of parameters including clinical findings, laboratory findings (nonspecific laboratory parameters such as eosinophilia, muscle enzymes, and serology), and epidemiological investigations. New regulations laying down rules for official controls for Trichinella in meat in order to improve food safety for consumers have recently been released in Europe. The evidence that the disease can be monitored and to some extent controlled with a rigorous reporting and testing system in place should be motivation to expand appropriate programs worldwide. PMID- 19136436 TI - Neuropathogenesis of congenital cytomegalovirus infection: disease mechanisms and prospects for intervention. AB - Congenital cytomegalovirus (CMV) infection is the leading infectious cause of mental retardation and hearing loss in the developed world. In recent years, there has been an improved understanding of the epidemiology, pathogenesis, and long-term disabilities associated with CMV infection. In this review, current concepts regarding the pathogenesis of neurological injury caused by CMV infections acquired by the developing fetus are summarized. The pathogenesis of CMV-induced disabilities is considered in the context of the epidemiology of CMV infection in pregnant women and newborn infants, and the clinical manifestations of brain injury are reviewed. The prospects for intervention, including antiviral therapies and vaccines, are summarized. Priorities for future research are suggested to improve the understanding of this common and disabling illness of infancy. PMID- 19136438 TI - Update on the human broad tapeworm (genus diphyllobothrium), including clinical relevance. AB - SUMMARY: Tapeworms (Cestoda) continue to be an important cause of morbidity in humans worldwide. Diphyllobothriosis, a human disease caused by tapeworms of the genus Diphyllobothrium, is the most important fish-borne zoonosis caused by a cestode parasite. Up to 20 million humans are estimated to be infected worldwide. Besides humans, definitive hosts of Diphyllobothrium include piscivorous birds and mammals, which represent a significant zoonotic reservoir. The second intermediate hosts include both freshwater and marine fish, especially anadromous species such as salmonids. The zoonosis occurs most commonly in countries where the consumption of raw or marinated fish is a frequent practice. Due to the increasing popularity of dishes utilizing uncooked fish, numerous cases of human infections have appeared recently, even in the most developed countries. As many as 14 valid species of Diphyllobothrium can cause human diphyllobothriosis, with D. latum and D. nihonkaiense being the most important pathogens. In this paper, all taxa from humans reported are reviewed, with brief information on their life history and their current distribution. Data on diagnostics, epidemiology, clinical relevance, and control of the disease are also summarized. The importance of reliable identification of human-infecting species with molecular tools (sequences of mitochondrial genes) as well as the necessity of epidemiological studies aimed at determining the sources of infections are pointed out. PMID- 19136440 TI - Empowerment evaluation with programs designed to prevent first-time male perpetration of sexual violence. AB - This special issue captures several threads in the ongoing evolution of sexual violence prevention. The articles that follow examine an empowerment evaluation process with four promising programs dedicated to preventing first-time male perpetration of sexual violence, as well as evaluation findings. Both the evaluation approach and the programs examined shed light on how sexual violence prevention can continue to be improved in the future. PMID- 19136441 TI - Empowerment evaluation for violence prevention public health programs. AB - The formative evaluation consultation and systems technique (FORECAST) approach to evaluating complex community programs is described as an empowerment evaluation strategy. First, empowerment evaluation is defined and contrasted with more-traditional approaches, such as experimental and quasi-experimental designs. Then FORECAST is described, illustrating how it was applied in four community projects. One of the projects is used to demonstrate the application of FORECAST evaluation in programs addressing the prevention of first-time male perpetration of sexual violence. PMID- 19136442 TI - Factors contributing to the effectiveness of four school-based sexual violence interventions. AB - This study extends past research by examining factors associated with changes in attitudes, knowledge, and intended behaviors related to sexual assault. This study included 1,182 participants from four unique multiple-session school-based sexual violence interventions. Implementation and participant factors examined include single- versus mixed-gender groups, group setting versus classroom lecture setting, and participant gender. Participants completed self administered, paper-and-pencil pre- and postsurveys. A significant desired overall effect was found on participants' reports of positive attitudes, beliefs, and behavior regarding sexual harassment and personal boundaries and positive dating relationship norms (from pretest to posttest). There were steeper increases over time in both measures, with larger mixed-gender/single-gender differences among boys than among girls. Differences in the impact of participating in mixed- versus single-gender groups depended on classroom versus small group settings. The implications of these findings are discussed for sexual assault prevention programs. PMID- 19136443 TI - Logic models as a tool for sexual violence prevention program development. AB - Sexual violence is a growing public health problem, and there is an urgent need to develop sexual violence prevention programs. Logic models have emerged as a vital tool in program development. The Centers for Disease Control and Prevention funded an empowerment evaluation designed to work with programs focused on the prevention of first-time male perpetration of sexual violence, and it included as one of its goals, the development of program logic models. Two case studies are presented that describe how significant positive changes can be made to programs as a result of their developing logic models that accurately describe desired outcomes. The first case study describes how the logic model development process made an organization aware of the importance of a program's environmental context for program success; the second case study demonstrates how developing a program logic model can elucidate gaps in organizational programming and suggest ways to close those gaps. PMID- 19136444 TI - Empowering programs with evaluation technical assistance: outcomes and lessons learned. AB - Although empowerment evaluation has gained widespread currency, few reports have described its outcomes. This article combines perspectives of participants and technical assistance providers to describe the process and outcomes of the Evaluation Assistance for Sexual Violence Programs project. Participating programs reported substantial enhancements in evaluation capacity, resources devoted to evaluation, and the extent and sophistication of their evaluation practice, as well as numerous examples of the application of evaluation findings to program improvement. Experiences from evaluation technical assistance providers identified aspects of the process that were particularly useful in achieving these outcomes, including investing in collaborative relationships, maximizing participation among program staff, tailoring the content and form of technical assistance to program preferences, and combining structured learning with program-specific technical assistance. PMID- 19136445 TI - "Like a family but better because you can actually trust each other": the Expect Respect dating violence prevention program for at-risk youth. AB - Expect Respect, a teen dating violence prevention program, was among four programs selected by the Centers for Disease Control and Prevention to participate in an empowerment evaluation project. As one aspect of this project, a qualitative study was designed to investigate the effects of Expect Respect support groups for at-risk youth. The goal was to understand the "how and why" of the program's impact on participants. Group interviews were conducted with five boys' and five girls' support groups after completion of the program. Settings included public middle and high schools and alternative schools in juvenile detention. Participants were asked to describe significant learning experiences in support groups as well as changes in their relationships resulting from program participation. Youths across all groups reported learning new skills including improved communication, anger control, and alternatives to violence. They reported increased knowledge about healthy relationships and warning signs of dating violence and expanded awareness of their own and others' abusive behaviors. Changed relationship norms were uniquely expressed by a boys' group in juvenile detention. Findings indicate that the experience of emotional safety in groups and positive relationships among group members were instrumental in the learning process. PMID- 19136446 TI - Adoption, adaptation, and fidelity of implementation of sexual violence prevention programs. AB - Little research examines the organizational and contextual dynamics that affect decisions to adopt evidence-based programs as well as the feasibility of implementation with fidelity to the original model when new users adopt established programs. To understand how promising strategies can be disseminated widely, this study examines the adoption and implementation of two sexual violence prevention programs in new settings. Interviews were conducted with stake-holders to investigate the factors and dynamics related to the adoption and implementation of these programs. Additionally, the research team worked with the program developers to create measures of the fidelity of implementation, which were then administered at each site. The findings suggest that adoption decisions were based on perceived fit between the program and the adopting organization's values, goals, and local setting. After adoption, new sites were able to implement the program with fairly high levels of fidelity, given moderate investments in training and technical assistance. PMID- 19136439 TI - AmpC beta-lactamases. AB - SUMMARY: AmpC beta-lactamases are clinically important cephalosporinases encoded on the chromosomes of many of the Enterobacteriaceae and a few other organisms, where they mediate resistance to cephalothin, cefazolin, cefoxitin, most penicillins, and beta-lactamase inhibitor-beta-lactam combinations. In many bacteria, AmpC enzymes are inducible and can be expressed at high levels by mutation. Overexpression confers resistance to broad-spectrum cephalosporins including cefotaxime, ceftazidime, and ceftriaxone and is a problem especially in infections due to Enterobacter aerogenes and Enterobacter cloacae, where an isolate initially susceptible to these agents may become resistant upon therapy. Transmissible plasmids have acquired genes for AmpC enzymes, which consequently can now appear in bacteria lacking or poorly expressing a chromosomal bla(AmpC) gene, such as Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis. Resistance due to plasmid-mediated AmpC enzymes is less common than extended spectrum beta-lactamase production in most parts of the world but may be both harder to detect and broader in spectrum. AmpC enzymes encoded by both chromosomal and plasmid genes are also evolving to hydrolyze broad-spectrum cephalosporins more efficiently. Techniques to identify AmpC beta-lactamase producing isolates are available but are still evolving and are not yet optimized for the clinical laboratory, which probably now underestimates this resistance mechanism. Carbapenems can usually be used to treat infections due to AmpC producing bacteria, but carbapenem resistance can arise in some organisms by mutations that reduce influx (outer membrane porin loss) or enhance efflux (efflux pump activation). PMID- 19136447 TI - Empowerment evaluation: building prevention science and organizational capacity to prevent sexual violence. AB - The articles in this special issue all result from the employment of empowerment evaluation with the intention of furthering the field of sexual violence prevention. Centers for Disease Control and Prevention's goals were to build the science toward evidence-based programming and to build evaluation capacity among leaders in the sexual violence field. Other undefined benefits also resulted, including the facilitation of implementation of program improvements and the development of capacities that can be generalized across other aspects of the participants' work. These benefits are attributed to empowerment evaluation's intentional designation of evaluation activities to the same people responsible for program delivery. Empowerment evaluation's potential to make future contributions to the field of sexual violence prevention is discussed. PMID- 19136448 TI - Bias in 2-part mixed models for longitudinal semicontinuous data. AB - Semicontinuous data in the form of a mixture of zeros and continuously distributed positive values frequently arise in biomedical research. Two-part mixed models with correlated random effects are an attractive approach to characterize the complex structure of longitudinal semicontinuous data. In practice, however, an independence assumption about random effects in these models may often be made for convenience and computational feasibility. In this article, we show that bias can be induced for regression coefficients when random effects are truly correlated but misspecified as independent in a 2-part mixed model. Paralleling work on bias under nonignorable missingness within a shared parameter model, we derive and investigate the asymptotic bias in selected settings for misspecified 2-part mixed models. The performance of these models in practice is further evaluated using Monte Carlo simulations. Additionally, the potential bias is investigated when artificial zeros, due to left censoring from some detection or measuring limit, are incorporated. To illustrate, we fit different 2-part mixed models to the data from the University of Toronto Psoriatic Arthritis Clinic, the aim being to examine whether there are differential effects of disease activity and damage on physical functioning as measured by the health assessment questionnaire scores over the course of psoriatic arthritis. Some practical issues on variance component estimation revealed through this data analysis are considered. PMID- 19136449 TI - Treatment of hemodialyzed patient with sunitinib. PMID- 19136450 TI - Sequence analysis and polymorphism discovery in 4 members of the bovine cathelicidin gene family. AB - Cathelicidins (CATHLs) are small, cationic antimicrobial peptides that establish an early innate immune defense against infections in mammals. Beyond their wide spectrum of antimicrobial activity, these peptides play important roles in wound repair, chemotactic activity, and apoptosis. Thus, polymorphisms present in bovine CATHLs 2, 5, 6, and 7 could potentially underlie inherited differences in innate immunity and disease resistance. The purpose of the present study was to characterize single nucleotide polymorphisms (SNPs) and insertion-deletion (indel) polymorphisms within the bovine CATHL gene family. Comparative sequence analysis for 10 domestic cattle breeds representing both Bos taurus and Bos indicus revealed 60 SNPs, 7 of which were nonsynonymous and 5 indel mutations. Characterization of these novel polymorphisms is central to developing a firm understanding regarding what effects, if any, nonsynonymous CATHL variation has with respect to bovine innate immunity. PMID- 19136451 TI - Divergence in the face of gene flow: the case of two newts (amphibia: salamandridae). AB - Understanding the process of divergence requires the quantitative characterization of patterns of gene flow between diverging taxa. New and powerful coalescent-based methods give insight into these processes in unprecedented details by enabling the reconstruction of the temporal distribution of past gene flow. Here, we use sequence variation at eight nuclear markers and mitochondrial DNA (mtDNA) in multiple populations to study diversity, divergence, and gene flow between two subspecies of a salamander, the smooth newt (Lissotriton vulgaris kosswigi and Lissotriton vulgaris vulgaris) in Turkey. The ranges of both subspecies encompass mainly the areas of this important glacial refugial area. Populations in refugia where species have been present for a long time and differentiated in situ should better preserve the record of past gene flow than young populations in postglacial expansion areas. Sequence diversity in both subspecies was substantial (nuclear pi(sil) = 0.69% and 1.31%). We detected long-term demographic stability in these refugial populations with large effective population sizes (N(e)) of the order of 1.5-3 x 10(5) individuals. Gene trees and the isolation with migration (IM) analysis complemented by tests of nested IM models showed that despite deep, pre-Pleistocene divergence of the studied newts, asymmetric introgression from vulgaris to kosswigi has occurred, with signatures of recent gene flow in mtDNA and an anonymous nuclear marker, and evidence for more ancient introgression in nuclear introns. The distribution of migration times raises the intriguing possibility that even the initial divergence may have occurred in the face of gene flow. PMID- 19136452 TI - Answer to the commentary: Politics and public health--some conceptual considerations concerning welfare state characteristics and public health outcomes. PMID- 19136454 TI - Hemispheric asymmetry of auditory evoked fields elicited by spectral versus temporal stimulus change. AB - The investigation of functional hemispheric asymmetries regarding auditory processing in the human brain still remains a challenge. Classical lesion and recent neuroimaging studies indicated that speech is dominantly processed in the left hemisphere, whereas music is dominantly processed in the right. However, recent studies demonstrated that the functional hemispheric asymmetries were not limited to the processing of highly cognitive sound signals like speech and music but rather originated from the basic neural processing of elementary sound features, that is, spectral and temporal acoustic features. Here, in contrast to previous studies, we used carefully composed tones and pulse trains as stimuli, balanced the overall physical sound input between spectral and temporal change conditions, and demonstrated the time course of neural activity evoked by spectral versus temporal sound input change by means of magnetoencephalography (MEG). These original findings support the hypothesis that spectral change is dominantly processed in the right hemisphere, whereas temporal change is dominantly processed in the left. PMID- 19136453 TI - Valproic acid-induced deregulation in vitro of genes associated in vivo with neural tube defects. AB - The utility of an in vitro system to search for molecular targets and markers of developmental toxicity was explored, using microarrays to detect genes susceptible to deregulation by the teratogen valproic acid (VPA) in the pluripotent mouse embryonal carcinoma cell line P19. Total RNA extracted from P19 cells cultured in the absence or presence of 1, 2.5, or 10mM VPA for 1.5, 6, or 24 h was subjected to replicated microarray analysis, using CodeLink UniSet I Mouse 20K Expression Bioarrays. A moderated F-test revealed a significant VPA response for 2972 (p < 10(-3)) array probes (19.4% of the filtered gene list), 421 of which were significant across all time points. In a core subset of VPA target genes whose expression was downregulated (68 genes) or upregulated (125 genes) with high probability (p < 10(-7)) after both 1.5 and 6 h of VPA exposure, there was a significant enrichment of the biological process Gene Ontology term transcriptional regulation among downregulated genes, and apoptosis among upregulated, and two of the downregulated genes (Folr1 and Gtf2i) have a knockout phenotype comprising exencephaly, the major malformation induced by VPA in mice. The VPA-induced gene expression response in P19 cells indicated that approximately 30% of the approximately 200 genes known from genetic mouse models to be associated with neural tube defects may be potential VPA targets, suggestive of a combined deregulation of multiple genes as a possible mechanism of VPA teratogenicity. Gene expression responses related to other known effects of VPA (histone deacetylase inhibition, G(1)-phase cell cycle arrest, induction of apoptosis) were also identified. This study indicates that toxicogenomic responses to a teratogenic compound in vitro may correlate with known in vitro and in vivo effects, and that short-time (< or =6 h) exposures in such an in vitro system could provide a useful component in mechanistic studies and screening tests in developmental toxicology. PMID- 19136455 TI - The effect of surgery for symptomatic endometriosis: the other side of the story. AB - BACKGROUND: Surgery is often considered the best treatment option in women with symptomatic endometriosis. However, extent and duration of the therapeutic benefit are still poorly defined. METHODS: The best available evidence on surgery for endometriosis-associated pain has been reviewed to estimate the effect size of interventions in the most frequently encountered clinical conditions. RESULTS: Methodological drawbacks limit considerably the validity of observational, non comparative studies on the effect of laparoscopy for stage I-IV disease. As indicated by the results of three RCTs, the absolute benefit increase of destruction of lesions compared with diagnostic only operation in terms of proportion of women reporting pain relief was between 30% and 40% after short follow-up periods. The effect size tended to decrease with time and the re operation rate, based on long-term follow-up studies, was as high as 50%. In most case series on excisional surgery for rectovaginal endometriosis, substantial short-term pain relief was experienced by approximately 70-80% of the subjects who continued the study. However, at 1 year follow-up, approximately 50% of the women needed analgesics or hormonal treatments. Major complications were observed in 3-10% of the patients. Medium-term recurrence of lesions was observed in approximately 20% of the cases, and around 25% of the women underwent repetitive surgery. CONCLUSIONS: Pain recurrence and re-operation rates after conservative surgery for symptomatic endometriosis are high and probably underestimated. Clinicians and patients should be aware that the expected benefit is operator dependent. PMID- 19136457 TI - Reproductive outcome after bariatric surgery: a critical review. AB - BACKGROUND: After many cycles of weight loss and weight gain, more and more morbidly obese patients undergo bariatric surgery, like gastric banding or gastric bypass, as the ultimate treatment for their obesity-problem. Since women of reproductive age are candidates for bariatric surgery, concerns arise regarding the potential impact on future pregnancy. METHODS: English-language articles were identified in a PUBMED search from 1982 to January 2008 using the keywords for pregnancy and bariatric surgery or gastric bypass or gastric banding. RESULTS: The few reported case-control and cohort studies clearly show improved fertility and a reduced risk in obstetrical complications, including gestational diabetes, macrosomia and hypertensive disorders of pregnancy, in women after operatively induced weight loss when compared with morbidly obesity women. The incidence of intrauterine growth restriction (IUGR) appears to be increased, however. No conclusions can be drawn concerning the risk for preterm labour and miscarriage, although these risks are probably increased compared with controls matched for body mass index. Operative complications are not uncommon with bariatric surgery and several cases have pointed to the increased risk for intestinal hernias and nutritional deficiencies in subsequent pregnancy. Deficiencies in iron, vitamin A, vitamin B(12), vitamin K, folate and calcium can result in both maternal complications, such as severe anaemia, and fetal complications, such as congenital abnormalities, IUGR and failure to thrive. CONCLUSIONS: Close supervision before, during and after pregnancy following bariatric surgery and nutrient supplementation adapted to the patient's individual requirements can help to prevent nutrition-related complications and improve maternal and fetal health, in this high-risk obstetric population. PMID- 19136456 TI - New insights into epididymal biology and function. AB - BACKGROUND: The epididymis performs an important role in the maturation of spermatozoa including their acquisition of progressive motility and fertilizing ability. However, the molecular mechanisms that govern these maturational events are still poorly defined. This review focuses on recent progress in our understanding of epididymal function including its development, role of the luminal microenvironment in sperm maturation, regulation and novel mechanisms the epididymis utilizes to carry out some of its functions. METHODS: A systematic search of Pubmed was carried out using the search term 'epididymis'. Articles that were published in the English language until the end of August 2008 and that focused on the specific topics described above were included. Additional papers cited in the primary reference were also included. RESULTS: While the majority of these findings were the result of studies in animal models, recent studies in the human epididymis are also presented including gene profiling studies to examine regionalized expression in normal epididymides as well as in those from vasectomized patients. CONCLUSIONS: Significant progress has been made in our understanding of epididymal function providing new insights that ultimately could improve human health. The data also indicate that the human epididymis plays an important role in sperm maturation but has unique properties compared with animal models. PMID- 19136458 TI - Influence of substituent modifications on the binding of 2-amino-1,8 naphthyridines to cytosine opposite an AP site in DNA duplexes: thermodynamic characterization. AB - Here, we report on a significant effect of substitutions on the binding affinity of a series of 2-amino-1,8-naphthyridines, i.e., 2-amino-1,8-naphthyridine (AND), 2-amino-7-methyl-1,8-naphthyridine (AMND), 2-amino-5,7-dimethyl-1,8-naphthyridine (ADMND) and 2-amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND), all of which can bind to cytosine opposite an AP site in DNA duplexes. Fluorescence titration experiments show that the binding affinity for cytosine is effectively enhanced by the introduction of methyl groups to the naphthyridine ring, and the 1:1 binding constant (10(6) M(-1)) follows in the order of AND (0.30) < AMND (2.7) < ADMND (6.1) < ATMND (19) in solutions containing 110 mM Na(+) (pH 7.0, at 20 degrees C). The thermodynamic parameters obtained by isothermal titration calorimetry experiments indicate that the introduction of methyl groups effectively reduces the loss of binding entropy, which is indeed responsible for the increase in the binding affinity. The heat capacity change (DeltaC(p)), as determined from temperature dependence of the binding enthalpy, is found to be significantly different between AND (-161 cal/mol K) and ATMND (-217 cal/mol K). The hydrophobic contribution appears to be a key force to explain the observed effect of substitutions on the binding affinity when the observed binding free energy (DeltaG(obs)) is dissected into its component terms. PMID- 19136459 TI - A bridged nucleic acid, 2',4'-BNA COC: synthesis of fully modified oligonucleotides bearing thymine, 5-methylcytosine, adenine and guanine 2',4'-BNA COC monomers and RNA-selective nucleic-acid recognition. AB - Recently, we synthesized pyrimidine derivatives of the 2'-O,4'-C methylenoxymethylene-bridged nucleic-acid (2',4'-BNA(COC)) monomer, the sugar conformation of which is restricted in N-type conformation by a seven-membered bridged structure. Oligonucleotides (BNA(COC)) containing this monomer show high affinity with complementary single-stranded RNA and significant resistance to nuclease degradation. Here, BNA(COC) consisting of 2',4'-BNA(COC) monomers bearing all four bases, namely thymine, 5-methylcytosine, adenine and guanine was efficiently synthesized and properties of duplexes containing the 2',4'-BNA(COC) monomers were investigated by UV melting experiments and circular dichroism (CD) spectroscopy. The UV melting curve analyses showed that the BNA(COC)/BNA(COC) duplex possessed excellent thermal stability and that the BNA(COC) increased thermal stability with a complementary RNA strand. On the other hand, BNA(COC)/DNA heteroduplexes showed almost the same thermal stability as RNA/DNA heteroduplexes. Furthermore, mismatched sequence studies showed that BNA(COC) generally improved the sequence selectivity with Watson-Crick base-pairing compared to the corresponding natural DNA and RNA. A CD spectroscopic analysis indicated that the BNA(COC) formed duplexes with complementary DNA and RNA in a manner similar to natural RNA. PMID- 19136460 TI - JBP1 and JBP2 are two distinct thymidine hydroxylases involved in J biosynthesis in genomic DNA of African trypanosomes. AB - Genomic DNA of African trypanosomes contains a hypermodified thymidine residue termed base J (beta-d-glucosyl-HOMedU). This modified base is localized primarily to repetitive DNA, namely the telomeres, and is implicated in the regulation of antigenic variation. The base is synthesized in a two-step pathway. Initially, a thymidine residue in DNA is hydroxylated by a thymidine hydroxylase (TH). This intermediate (HOMedU) is then glucosylated to form base J. Two proteins involved in J synthesis, JBP1 (J binding protein 1) and JBP2, contain a putative TH domain related to the family of Fe(2+)/2-oxoglutarate-dependent hydroxylases. We have previously shown that mutations in the TH domain of JBP1 kill its ability to stimulate J synthesis. Here we show that mutation of key residues in the TH domain of JBP2 ablate its ability to induce de novo J synthesis. While the individual JBP1 null and JBP2 null trypanosomes have reduced J levels, the deletion of both JBP1 and JBP2 generates a cell line that completely lacks base J but still contains glucosyl-transferase activity. Reintroduction of JBP2 in the J null trypanosome stimulates HOMedU formation and site-specific synthesis of base J. We conclude that JBP2 and JBP1 are the TH enzymes involved in J biosynthesis. PMID- 19136461 TI - Binding to DNA of the RNA-polymerase II C-terminal domain allows discrimination between Cdk7 and Cdk9 phosphorylation. AB - The C-terminal domain (CTD) of RNA polymerase II regulates transcription through spatially and temporally coordinated events. Previous work had established that the CTD binds DNA but the significance of this interaction has not been determined. The present work shows that the CTD binds DNA in its unphosphorylated form, the form in which it is present in the pre-initiation complex. The CTD/DNA complex is recognized by and is phosphorylated by Cdk7 but not by Cdk9. Model building studies indicate the structural mechanism underlying such specificity involves interaction of Cdk7 with DNA in the context of the CTD/DNA complex. The model has been tested by mutagenesis experiments. CTD dissociates from DNA following phosphorylation by Cdk7, allowing transcription initiation. The CTD then becomes accessible for further phosphorylation by Cdk9 that drives the transition to transcription elongation. PMID- 19136462 TI - Differential usage of alternate promoters of the human stress response gene ATF3 in stress response and cancer cells. AB - Stress response gene ATF3 plays a pleiotropic role in determining cell fate in response to mitogenic or stress stimuli. An alternate promoter of the human ATF3 gene (designated P1 in this study) has recently been reported, which is located approximately 43.5 kb upstream of the previously reported P2 promoter. We showed here that the P1 promoter is highly conserved between human and mouse and is functional in response to various stimuli, whereas the P1 promoter was dominantly induced by serum and the P2 promoter was more efficiently activated in response to TGF-beta and oncogenic HRAS. The P1 promoter contains multiple transcriptional start sites, and the different 5'-UTRs markedly affected their translation in response to stress. In human prostate and Hodgkin Reed-Sternberg cancer cells with elevated expression of ATF3, the P1 promoter was constitutively activated and its chromatin structure was modified into active configuration. The differential usage of alternate promoters of the ATF3 gene at both transcriptional and translational level and the modification of chromatin structure may provide a novel mechanism for expressing ATF3 in determining cell fate during stress response and cancer. PMID- 19136463 TI - Insights into the architecture and stoichiometry of Escherichia coli PepA*DNA complexes involved in transcriptional control and site-specific DNA recombination by atomic force microscopy. AB - Multifunctional Aminopeptidase A (PepA) from Escherichia coli is involved in the control of two distinct DNA transaction processes: transcriptional repression of the carAB operon, encoding carbamoyl phosphate synthase and site-specific resolution of ColE1-type plasmid multimers. Both processes require communication at a distance along a DNA molecule and PepA is the major structural component of the nucleoprotein complexes that underlie this communication. Atomic Force Microscopy was used to analyze the architecture of PepA.carAB and PepA.cer site complexes. Contour length measurements, bending angle analyses and volume determinations demonstrate that the carP1 operator is foreshortened by approximately 235 bp through wrapping around one PepA hexamer. The highly deformed part of the operator extends from slightly upstream of the -35 hexamer of the carP1 promoter to just downstream of the IHF-binding site, and comprises the binding sites for the PurR and RutR transcriptional regulators. This extreme remodeling of the carP1 control region provides a straightforward explanation for the strict requirement of PepA in the establishment of pyrimidine and purine specific repression of carAB transcription. We further provide a direct physical proof that PepA is able to synapse two cer sites in direct repeat in a large interwrapped nucleoprotein complex, likely comprising two PepA hexamers. PMID- 19136464 TI - Genetic screening for modifiers of the DREF pathway in Drosophila melanogaster: identification and characterization of HP6 as a novel target of DREF. AB - The DNA replication-related element-binding factor (DREF) regulates cell proliferation-related gene expression in Drosophila. By genetic screening, taking advantage of the rough eye phenotype of transgenic flies that express DREF in the eye discs, we identified 24 genes that suppressed and 12 genes that enhanced the rough eye phenotype when heterozygous for mutations. Five genes, HP6, pigeon, lace, X box binding protein 1 and guftagu were found to carry replication-related element (DRE) sequences in their 5'-flanking regions. Of these, the HP6 gene carries two sequences that match seven out of eight nucleotides of DRE and two additional sequences that match six out of eight nucleotides of DRE in the 5' flanking region. Band mobility shift assays using Drosophila Kc cell nuclear extracts demonstrated DREF binding to two of these sites and chromatin immunoprecipitation using anti-DREF antibodies confirmed that this occurs in vivo. Knockdown of DREF in Drosophila S2 cells decreased the HP6 mRNA level. The results, taken together, indicate that DREF directly regulates expression of the HP6 gene. HP6 mRNA was detected throughout development by RT-PCR with highest levels in adult males. In addition, immunostaining analyses revealed colocalization of HP6 and DREF in nuclei at the apical tips in the testes. PMID- 19136465 TI - A single anti-microRNA antisense oligodeoxyribonucleotide (AMO) targeting multiple microRNAs offers an improved approach for microRNA interference. AB - Anti-miRNA antisense inhibitors (AMOs) have demonstrated their utility in miRNA research and potential in miRNA therapy. Here we report a modified AMO approach in which multiple antisense units are engineered into a single unit that is able to simultaneously silence multiple-target miRNAs, the multiple-target AMO or MTg AMO. We validated the technique with two separate MTg-AMOs: anti-miR-21/anti-miR 155/anti-miR-17-5p and anti-miR-1/anti-miR-133. We first verified the ability of the MTg-AMOs to antagonize the repressive actions of their target miRNAs using luciferase reporter activity assays and to specifically knock down the levels of their target miRNAs using real-time RT-PCR methods. We then used the MTg-AMO approach to identify several tumor suppressors-TGFBI, APC and BCL2L11 as the target genes for oncogenic miR-21, miR-155 and miR-17-5p, respectively, and two cardiac ion channel genes HCN2 (encoding a subunit of cardiac pacemaker channel) and CACNA1C (encoding the alpha-subunit of cardiac L-type Ca(2+) channel) for the muscle-specific miR-1 and miR-133. We further demonstrated that the MTg-AMO targeting miR-21, miR-155 and miR-17-5p produced a greater inhibitory effect on cancer cell growth, compared with the regular single-target AMOs. Moreover, while using the regular single-target AMOs excluded HCN2 as a target gene for either miR-1 or miR-133, the MTg-AMO approach is able to reveal HCN2 as the target for both miR-1 and miR-133. Our findings suggest the MTg-AMO as an improved approach for miRNA target finding and for studying function of miRNAs. This approach may find its broad application for exploring biological processes involving multiple miRNAs and multiple genes. PMID- 19136466 TI - FRAXE-associated mental retardation protein (FMR2) is an RNA-binding protein with high affinity for G-quartet RNA forming structure. AB - FRAXE is a form of mild to moderate mental retardation due to the silencing of the FMR2 gene. The cellular function of FMR2 protein is presently unknown. By analogy with its homologue AF4, FMR2 was supposed to have a role in transcriptional regulation, but robust evidences supporting this hypothesis are lacking. We observed that FMR2 co-localizes with the splicing factor SC35 in nuclear speckles, the nuclear regions where splicing factors are concentrated, assembled and modified. Similarly to what was reported for splicing factors, blocking splicing or transcription leads to the accumulation of FMR2 in enlarged, rounded speckles. FMR2 is also localized in the nucleolus when splicing is blocked. We show here that FMR2 is able to specifically bind the G-quartet forming RNA structure with high affinity. Remarkably, in vivo, in the presence of FMR2, the ESE action of the G-quartet situated in mRNA of an alternatively spliced exon of a minigene or of the putative target FMR1 appears reduced. Interestingly, FMR1 is silenced in the fragile X syndrome, another form of mental retardation. All together, our findings strongly suggest that FMR2 is an RNA binding protein, which might be involved in alternative splicing regulation through an interaction with G-quartet RNA structure. PMID- 19136467 TI - Structure and expression analysis of rice paleo duplications. AB - Having a well-known history of genome duplication, rice is a good model for studying structural and functional evolution of paleo duplications. Improved sequence alignment criteria were used to characterize 10 major chromosome-to chromosome duplication relationships associated with 1440 paralogous pairs, covering 47.8% of the rice genome, with 12.6% of genes that are conserved within sister blocks. Using a micro-array experiment, a genome-wide expression map has been produced, in which 2382 genes show significant differences of expression in root, leaf and grain. By integrating both structural (1440 paralogous pairs) and functional information (2382 differentially expressed genes), we identified 115 paralogous gene pairs for which at least one copy is differentially expressed in one of the three tissues. A vast majority of the 115 paralogous gene pairs have been neofunctionalized or subfunctionalized as 88%, 89% and 96% of duplicates, respectively, expressed in grain, leaf and root show distinct expression patterns. On the basis of a Gene Ontology analysis, we have identified and characterized the gene families that have been structurally and functionally preferentially retained in the duplication showing that the vast majority (>85%) of duplicated have been either lost or have been subfunctionalized or neofunctionalized during 50-70 million years of evolution. PMID- 19136468 TI - Passport, a native Tc1 transposon from flatfish, is functionally active in vertebrate cells. AB - The Tc1/mariner family of DNA transposons is widespread across fungal, plant and animal kingdoms, and thought to contribute to the evolution of their host genomes. To date, an active Tc1 transposon has not been identified within the native genome of a vertebrate. We demonstrate that Passport, a native transposon isolated from a fish (Pleuronectes platessa), is active in a variety of vertebrate cells. In transposition assays, we found that the Passport transposon system improved stable cellular transgenesis by 40-fold, has an apparent preference for insertion into genes, and is subject to overproduction inhibition like other Tc1 elements. Passport represents the first vertebrate Tc1 element described as both natively intact and functionally active, and given its restricted phylogenetic distribution, may be contemporaneously active. The Passport transposon system thus complements the available genetic tools for the manipulation of vertebrate genomes, and may provide a unique system for studying the infiltration of vertebrate genomes by Tc1 elements. PMID- 19136469 TI - Discovery of novel triple helical DNA intercalators by an integrated virtual and actual screening platform. AB - Virtual Screening is an increasingly attractive way to discover new small molecules with potential medicinal value. We introduce a novel strategy that integrates use of the molecular docking software Surflex with experimental validation by the method of competition dialysis. This integrated approach was used to identify ligands that selectively bind to the triplex DNA poly(dA) [poly(dT)](2). A library containing approximately 2 million ligands was virtually screened to identify compounds with chemical and structural similarity to a known triplex intercalator, the napthylquinoline MHQ-12. Further molecular docking studies using compounds with high structural similarity resulted in two compounds that were then demonstrated by competition dialysis to have a superior affinity and selectivity for the triplex nucleic acid than MHQ-12. One of the compounds has a different chemical backbone than MHQ-12, which demonstrates the ability of this strategy to 'scaffold hop' and to identify small molecules with novel binding properties. Biophysical characterization of these compounds by circular dichroism and thermal denaturation studies confirmed their binding mode and selectivity. These studies provide a proof-of-principle for our integrated screening strategy, and suggest that this platform may be extended to discover new compounds that target therapeutically relevant nucleic acid morphologies. PMID- 19136470 TI - A designed RNA selection: establishment of a stable complex between a target and selectant RNA via two coordinated interactions. AB - In this paper, we describe a new method for selecting RNA aptamers that cooperatively bind to two specific sites within a target RNA. We designed a selection system in which two RNAs, a target RNA and a RNA pool, were assembled by employing a pre-organized GAAA tetraloop-11-nt receptor interaction. This allows us to select the binding sequence against a targeted internal loop as well as a linker region optimized for binding of the two binding sites. After the selection, the aptamers bound with dissociation constants in the nanomolar range, thereby forming a stable complex with the target RNA. Thus this method enables identification of aptamers for a specific binding site together with a linker for cooperative binding of the two RNAs. It appears that our new method can be applied generally to select RNAs that adhere tightly to a target RNA via two specific sites. The method can also be applicable for further engineering of both natural and artificial RNAs. PMID- 19136475 TI - DNA repair-deficient Xpa/p53 knockout mice are sensitive to the non-genotoxic carcinogen cyclosporine A: escape of initiated cells from immunosurveillance? AB - The DNA repair-deficient Xpa(-/-)p53(+/-) (Xpa/p53) mouse is a potent model for carcinogenicity testing, representing increased sensitivity toward genotoxic but surprisingly also toward true human non-genotoxic carcinogens. The mechanism of this increased sensitivity in Xpa/p53 mice toward non-genotoxic carcinogens is still unknown. Here, we investigated the mechanism of the human non-genotoxic carcinogen cyclosporine A (CsA) in the Xpa/p53 mouse model. Xpa/p53 mice exposed to CsA for 39 weeks showed a significantly increased lymphoma incidence as compared with untreated Xpa/p53 mice and CsA-treated wild-type (WT) mice. We excluded concealed genotoxicity of CsA in Xpa/p53 mice by mutant frequency analyses. As a next step, we used a genetic approach: immunodeficient DNA-PKcs mice, defective in the catalytic subunit of the DNA-dependent protein kinase, were crossed with Xpa and Xpa/p53 mice. Xpa/p53 mice had an increased lymphoma incidence with shorter latency times as compared with DNA-PKcs-deficient WT and Xpa mice. Surprisingly, also six of 15 DNA-PKcs/Xpa/p53 females had developed an adenocarcinoma of the mammary gland. Tumor responses in CsA-treated and DNA-PKcs deficient Xpa/p53 mice were comparable as both genotypes developed mainly splenic lymphomas enriched in B lymphocytes. From our present studies, we hypothesize that levels of initiated precancerous cells are elevated in Xpa/p53 mice. These cells are insufficiently eliminated due to either suppression of the immune system by CsA or through immune-related DNA-PKcs deficiency. Based on the current studies and those conducted previously, we conclude that the Xpa/p53 model is an excellent adjunct to the current chronic rodent bioassay. PMID- 19136474 TI - Phosphoaspirin (MDC-43), a novel benzyl ester of aspirin, inhibits the growth of human cancer cell lines more potently than aspirin: a redox-dependent effect. AB - Aspirin is chemopreventive against colon and probably other cancers, but this effect is relatively weak and its chronic administration to humans is associated with significant side effects. Because of these limitations, extensive effort has been exerted to improve the pharmacological properties of aspirin. We have determined the anticancer activity and mechanisms of action of the novel para positional isomer of phosphoaspirin [P-ASA; MDC-43; 4 ((diethoxyphosphoryloxy)methyl)phenyl 2-acetoxybenzoate]. P-ASA inhibited the growth of 10 human cancer cell lines originating from colon, lung, liver, pancreas and breast, at least 18- to 144-fold more potently than conventional aspirin. P-ASA achieved this effect by modulating cell kinetics; compared with controls, P-ASA reduced cell proliferation by up to 68%, increased apoptosis 5.5 fold and blocked cell cycle progression in the G(2)/M phase. P-ASA increased intracellular levels of reactive oxygen species (ROS), depleted glutathione levels and modulated cell signaling predominantly through the mitogen-activated protein kinase (p38 and c-jun N-terminal kinase), cyclooxygenase (COX) and nuclear factor-kappa B pathways. P-ASA targeted the mitochondria, increasing mitochondrial superoxide anion levels; this effect on ROS led to collapsed mitochondrial membrane potential and triggered the intrinsic apoptotic pathway. The antioxidant N-acetyl cysteine abrogated the cell growth inhibitory and signaling effects of P-ASA, underscoring the centrality of ROS in its mechanism of action. Our results, establishing P-ASA as a potent inhibitor of the growth of several human cancer cell lines, suggest that it may possess broad anticancer properties. We conclude that the novel P-ASA is a promising anticancer agent, which merits further evaluation. PMID- 19136476 TI - 15-Deoxy-Delta12,14-prostaglandin J2 upregulates the expression of heme oxygenase 1 and subsequently matrix metalloproteinase-1 in human breast cancer cells: possible roles of iron and ROS. AB - Heme oxygenase-1 (HO-1) has recently been found to be involved in angiogenesis and metastasis. In this study, we investigated whether HO-1 could potentiate the metastatic potential of human breast cancer cells. Treatment of MCF-7 and MDA-MB 231 cells with 30 microM of 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) increased the expression of HO-1, which preceded the induction of matrix metalloproteinases (MMPs). The 15d-PGJ2-induced upregulation of MMP-1 was abrogated by the HO-1 inhibitor zinc protoporphyrin IX (ZnPP) as well as introduction of HO-1 short interfering RNA. In addition, HO-1 inducers, such as cobalt protoporphyrin IX and hemin, upregulated the expression of MMP-1. Overexpression of HO-1 in the MCF-7 cells caused the induction of MMP-1 expression. Treatment with the HO-1 inhibitor ZnPP abolished the migrative phenotype of 15d-PGJ2-treated MCF-7 cells. MCF-7 cells treated with 15d-PGJ2 exhibited intracellular accumulation of reactive oxygen species (ROS) which was abolished by ZnPP. We hypothesize that excess iron, released as a consequence HO 1 activity induced by 15d-PGJ2, is transiently available for the stimulation of intracellular ROS generation and subsequently MMP-1 expression. 15d-PGJ2-mediated upregulation of MMP-1 expression was blocked by the iron chelator desferrioxamine and the Fe2+-specific chelator 1,10-phenanthroline. The iron chelators as well as the antioxidant N-acetyl-L-cysteine abrogated ROS formation by 15d-PGJ2. In conclusion, 15d-PGJ2 upregulates MMP-1 expression via induction of HO-1 and subsequent production of iron capable of generating ROS, which may contribute to increased metastasis and invasiveness of the human breast cancer cells. PMID- 19136477 TI - The COX-2/PGE2 pathway: key roles in the hallmarks of cancer and adaptation to the tumour microenvironment. AB - It is widely accepted that alterations to cyclooxygenase-2 (COX-2) expression and the abundance of its enzymatic product prostaglandin E(2) (PGE(2)) have key roles in influencing the development of colorectal cancer. Deregulation of the COX 2/PGE(2) pathway appears to affect colorectal tumorigenesis via a number of distinct mechanisms: promoting tumour maintenance and progression, encouraging metastatic spread, and perhaps even participating in tumour initiation. Here, we review the role of COX-2/PGE(2) signalling in colorectal tumorigenesis and highlight its ability to influence the hallmarks of cancer--attributes defined by Hanahan and Weinberg as being requisite for tumorigenesis. In addition, we consider components of the COX-prostaglandin pathway emerging as important regulators of tumorigenesis; namely, the prostanoid (EP) receptors, 15 hydroxyprostaglandin dehydrogenase and the prostaglandin transporter. Finally, based on recent findings, we propose a model for the cellular adaptation to the hypoxic tumour microenvironment that encompasses the interplay between COX-2, hypoxia-inducible factor 1 and dynamic switches in beta-catenin function that fine-tune signalling networks to meet the ever-changing demands of a tumour. PMID- 19136478 TI - Macrophages and nerve fibres in peritoneal endometriosis. AB - BACKGROUND: Endometriosis is considered to be an inflammatory disease, and macrophages are the most numerous immune cells in endometriotic lesions. However, the mechanisms underlying the elevation of macrophages and their role in the pathogenesis and manifestations of endometriosis still remain unclear. METHODS: The number of macrophages stained for CD68 in endometriotic lesions (n = 24) and in peritoneum distant from the lesions (n = 14) from women with endometriosis was compared with the number of macrophages in normal peritoneum from women without endometriosis (n = 18). Peritoneal lesions were also double-stained for CD68 and protein gene product 9.5 to study the relationship between macrophages and nerve fibres. RESULTS: The densities of macrophages in peritoneal endometriotic lesions and unaffected peritoneum from women with endometriosis were both significantly higher than that in normal peritoneum from women without endometriosis (P < 0.001). More nerve fibres were also found in the areas where increased numbers of macrophages were identified. CONCLUSIONS: There was a significant elevation of macrophages in both normal peritoneum and peritoneal lesions from women with endometriosis compared with normal peritoneum from women without endometriosis. These cells may well play roles in the growth and development of endometriotic lesions and in the generation of pain through interaction with nerve fibres. PMID- 19136479 TI - Infertility patients' beliefs about their embryos and their disposition preferences. AB - BACKGROUND: Little research has been done on how beliefs of infertility patients about their embryos are related to their disposition decisions. The objective of this study was to describe how patients speak about their embryos, in moral (e.g. status) and non-moral terms, and to investigate how patients' narratives are related to their disposition preferences. METHODS: In-depth interviews based on the method of interpretative phenomenological analysis were conducted with patients undergoing IVF/ICSI treatment between May and July 2006. RESULTS: Seven couples and 11 female patients were interviewed. Six major themes emerged from the narratives of the participants when they spoke about their embryos: (i) a medical-technical perspective; (ii) feelings; (iii) genetic link to oneself and/or one's partner; (iv) symbolic meaning of the relationship between the infertile partners; (v) moral status and (vi) instrumental value. All but two participants spontaneously considered the embryo disposition options as a two stage decision sequence. In the first step, they considered donation to another couple for reproductive purposes. At this stage, the presence of the themes 'genetic link' and 'symbol of the relationship' was linked with a clear reluctance to donate. In the second step of the decision-making process, the option of donation for research and discarding were considered. At this stage, participants' confidence in medical science and the instrumental value they attached to the embryo were related to their decisions. CONCLUSIONS: Patients' conceptualization of their embryos plays an important role in embryo disposition decisions. Our research showed that patients deal with these decisions in a two stage decision sequence. PMID- 19136480 TI - Prospective longitudinal cohort study on cumulative 5-year delivery and adoption rates among 1338 couples initiating infertility treatment. AB - BACKGROUND: The objective was to assess crude 5-year delivery rates after assisted reproductive technology (ART) treatment, intrauterine inseminations (IUI), spontaneous conceptions (SC) and adoptions in a large infertile cohort. METHODS: A prospective longitudinal survey comprised 1338 infertile couples starting public infertility programmes offering IUIs and three free ART cycles during 2000-2001. The cohort was cross-linked with the National Medical Birth Register to obtain delivery rates for all 1338 couples. More detailed data were available from 817 women responding to a 5-year follow-up questionnaire (response rate 74.7%). Fifty-seven percent (466/817) of the couples had received treatment prior to inclusion in the study with an average of 4.1+/-2.8 infertility treatments before referral. RESULTS: Of the 1338 couples, 69.4% had at least one delivery within 5-years of follow-up. For women <35 years 74.9% had delivered compared with 52.2% of those aged > or =35 years. The mean number of children was 1.6, and 52.1% had more than one child. Of the 817 women who provided questionnaire data, 18.2% (149/817) delivered after SC, two-thirds of these after a previous ART delivery. Adoption of a child occurred for 5.9% (48/817) of the women. Positive prognostic factors for delivery were male infertility, female age <35 years, <3 years of infertility and less than three previous treatment cycles. CONCLUSIONS: A crude delivery rate of 69.4% in the total population 5 years after referral to tertiary hospital centres with 6.6% deliveries after SC in the subpopulation responding to the questionnaire indicates a high efficacy of modern infertility treatments. PMID- 19136481 TI - Novel interventions to reduce re-infection in women with chlamydia: a randomized controlled trial. AB - BACKGROUND: The aim of this study was to determine whether postal testing kits (PTKs) or patient-delivered partner therapy (PDPT) for partners of women with Chlamydia trachomatis reduce re-infection rates in women, compared with partner notification by patient referral. METHODS: Three hundred and thirty women testing positive for chlamydia, at clinics for genitourinary medicine, family planning and termination of pregnancy in Edinburgh, were randomized to one of three partner interventions: patient referral, PTK (partners post urine for testing) or PDPT (1 g azithromycin for partners). Women submitted urine for chlamydia testing every 3 months. The primary outcome was re-infection assessed as time to first positive result by the Cox proportional hazard regression. The proportion of partners tested or treated with each intervention was determined. RESULTS: Out of 330 women, 215 (65%) were retested over 12 months. There were 32 of 215 women (15%) who retested positive (7, 15 and 10 women from the patient referral, PTK and PDPT groups, respectively). There was no significant difference in re infection between PDPT versus patient referral (HR 1.32, 95% CI 0.50-3.56), PTK versus patient referral (HR 2.35, 95% CI 0.94-5.88) or PDPT versus PTK (HR 0.55, 95% CI 0.24-1.24). There was no significant difference in the proportion of partners confirmed tested/treated between the patient referral (34%) and PTK (41%, P = 0.32) or PDPT (42%, P = 0.28) groups. CONCLUSIONS: PTK and PDPT do not reduce re-infection rates in women with chlamydia compared with patient referral. However, PDPT may offer other advantages such as simplicity and cost compared with patient referral. PMID- 19136482 TI - Biomarkers of pathophysiology in hypertrophic cardiomyopathy: implications for clinical management and prognosis. AB - The study of biomarkers and their signalling pathways has allowed the development of new therapeutic strategies in a range of disorders. The aim of the present systematic review is to provide an overview of different biomarkers in patients with hypertrophic cardiomyopathy that could give some insight into the pathophysiologic mechanism(s) underlying the typical clinical and histological manifestations of the disease. Several pathophysiological models are presented and discussed, including studies that have investigated these biomarkers for diagnostic and prognostic reasons, in relation to disease progression and/or mortality. PMID- 19136483 TI - Symptomatic travel associated pulmonary embolism: high severity does not imply poor long term prognosis. PMID- 19136484 TI - Patients with peripheral arterial disease in the CHARISMA trial. AB - AIMS: The aim of this study was to determine whether clopidogrel plus aspirin provides greater protection against major cardiovascular events than aspirin alone in patients with peripheral arterial disease (PAD). METHODS AND RESULTS: This is a post hoc analysis of the 3096 patients with symptomatic (2838) or asymptomatic (258) PAD from the CHARISMA trial. The rate of cardiovascular death, myocardial infarction (MI), or stroke (primary endpoint) was higher in patients with PAD than in those without PAD: 8.2% vs. 6.8% [hazard ratio (HR), 1.25; 95% CI 1.08, 1.44; P = 0.002]. Among the patients with PAD, the primary endpoint occurred in 7.6% in the clopidogrel plus aspirin group and 8.9% in the placebo plus aspirin group (HR, 0.85; 95% CI, 0.66-1.08; P = 0.18). In these patients, the rate of MI was lower in the dual antiplatelet arm than the aspirin alone arm: 2.3% vs. 3.7% (HR, 0.63; 95% CI, 0.42-0.96; P = 0.029), as was the rate of hospitalization for ischaemic events: 16.5% vs. 20.1% (HR, 0.81; 95% CI, 0.68 0.95; P = 0.011). The rates of severe, fatal, or moderate bleeding did not differ between the groups, whereas minor bleeding was increased with clopidogrel: 34.4% vs. 20.8% (odds ratio, 1.99; 95% CI, 1.69-2.34; P < 0.001). CONCLUSION: Dual therapy provided some benefit over aspirin alone in PAD patients for the rate of MI and the rate of hospitalization for ischaemic events, at the cost of an increase in minor bleeding. PMID- 19136485 TI - The peripheral arterial disease subgroup in the CHARISMA trial: does it tell us anything new? PMID- 19136487 TI - Validation of an echocardiographic multiparametric strategy to increase responders patients after cardiac resynchronization: a multicentre study. AB - AIMS: We sought to develop and validate a multiparametric algorithm by applying previously validated criteria to predict cardiac resynchronization therapy (CRT) response in a multicentre study. Thirty per cent of patients treated by CRT fail to respond to the treatment. Although dyssynchrony by echocardiography has been used to improve the selection of patients, the complexity of myocardial contraction has generated a moderate improvement using any of several individual parameters. METHODS AND RESULTS: Two hundred end-stage heart failure patients [NYHA 3-4 and left ventricular ejection fraction (LVEF)<35%] with QRS>120 ms were included. Echocardiography analysis focused on the following parameters: atrioventricular dyssynchrony, interventricular dyssynchrony, and intraventricular dyssynchrony that integrated radial (PSAX M-mode) and longitudinal [tissue Doppler imaging (TDI)] evaluations for spatial (wall to wall) and temporal (wall end-systole to mitral valve opening) dyssynchrony diagnosis. Following CRT implantation, patients were monitored for 6 months with functional and echo evaluations defining responders by a 15% reduction in end systolic volume. Mean QRS duration and LVEF were 152 +/- 17 ms and 25 +/- 8%. There was a CRT response in 57% of patients, independent of QRS width. Mean prevalence of positive criteria was 34 +/- 8%. Feasibility and variability averages were 81 +/- 20% and 9 +/- 4%. In a single parametric approach, ranges of sensitivities and specificities were 18-65% and 45-84% with a mean of 41% and 66%. A multiparametric approach by focusing on criteria combination decreased the mean rate of false-positive results to 14 +/- 12%, 5 +/- 4%, 2 +/- 2%, and 1 +/- 2% from one to four parameters, respectively. More than three parameters were associated with a specificity above 90% and a positive predictive value above 65%. Reproducibility of this global strategy was 91%. CONCLUSION: A multiparametric echocardiographic strategy based on the association of conventional criteria is a better indicator of CRT response than the existing single parametric approaches. PMID- 19136488 TI - Association of pericardial fat, intrathoracic fat, and visceral abdominal fat with cardiovascular disease burden: the Framingham Heart Study. AB - AIMS: The aim of this study was to assess whether pericardial fat, intrathoracic fat, and visceral abdominal adipose tissue (VAT) are associated with the prevalence of cardiovascular disease (CVD). METHODS AND RESULTS: Participants from the Framingham Heart Study Offspring cohort underwent abdominal and chest multidetector computed tomography to quantify volumes of pericardial fat, intrathoracic fat, and VAT. Relations between each fat depot and CVD were assessed using logistic regression. The analysis of 1267 participants (mean age 60 years, 53.8% women, 9.7% with prevalent CVD) demonstrated that pericardial fat [odds ratio (OR) 1.32, 95% confidence interval (CI) 1.11-1.57; P = 0.002] and VAT (OR 1.35, 95% CI 1.11-1.57; P = 0.003), but not intrathoracic fat (OR 1.14, 95% CI 0.93-1.39; P = 0.22), were significantly associated with prevalent CVD in age sex-adjusted models and after adjustment for body mass index and waist circumference. After multivariable adjustment, associations were attenuated (P > 0.14). Only pericardial fat was associated with prevalent myocardial infarction after adjusting for conventional measures of adiposity (OR 1.37, 95% CI 1.03 1.82; P = 0.03). CONCLUSION: Pericardial fat and VAT, but not intrathoracic fat, are associated with CVD independent of traditional measures of obesity but not after further adjustment for traditional risk factor. Taken together with our prior work, these findings may support the hypothesis that pericardial fat contributes to coronary atherosclerosis. PMID- 19136489 TI - Effect of right ventricular lead location on response to cardiac resynchronization therapy in patients with end-stage heart failure. AB - AIMS: It is currently recommended to implant the left ventricular (LV) pacing lead at the lateral wall. However, the optimal right ventricular (RV) pacing lead location for cardiac resynchronization therapy (CRT) remains controversial. We sought to investigate whether optimizing the site for placement of the RV lead could further improve the long-term response to CRT in patients with advanced heart failure. METHODS AND RESULTS: Between October 2006 and December 2007, a total of 73 consecutive patients with standard indication for CRT were enrolled. The enrolled patients were divided into two groups based on the RV lead location. There were 50 patients in RV apex (RVA) group and 23 patients in RV high septum (RVHS). The primary study endpoint was a decrease in LV end-systolic volume (LVESV) by >15% at 6-month follow-up. The secondary endpoints were improvement in New York Heart Association (NYHA) class by >or=1 point and decrease in brain-type natriuretic peptide (BNP) levels by >50% after CRT. At 6-month follow-up, improvement in NYHA class by >or=1 point (RVA: 72% vs. RVHS: 74%, P = 0.76), decrease in LVESV by >or=15% (RVA: 65% vs. RVHS: 64%, P = 0.76), and decrease in BNP level by >50% (RVA: 70% vs. RVHS: 69%, P = 0.88) were observed in similar proportion of the two groups. When we separately assessed the significance of RV pacing site in three LV stimulation sites, there were no significant differences in terms of clinical improvement (62 vs. 64%, P = 0.74) and decrease in LVESV by >15% (63 vs. 62%, P = 0.78) between RVA and RVHS pacing when the LV stimulation site was lateral cardiac vein. In anterolateral vein pacing site, the RVA stimulation was associated with higher clinical (88 vs. 47%, P = 0.05), echocardiographic (75 vs. 32%, P = 0.02), and neurohormonal responses (80 vs. 50%, P = 0.04) compared with that in RVHS site. When LV was paced from posterolateral vein, RVHS pacing was superior to RVA in terms of the clinical improvement (85 vs. 35%, P = 0.01), echocardiographic response (72 vs. 30%, P = 0.01), and decrease in BNP levels (75 vs. 50%, P = 0.04). CONCLUSION: The present study did not show any difference between RVA and RVHS pacing sites in terms of overall improvement in clinical outcome and LV reverse remodelling following CRT. However, effect of RV lead location on CRT response varies depending on LV stimulation site. PMID- 19136486 TI - Efficacy of the I(f) current inhibitor ivabradine in patients with chronic stable angina receiving beta-blocker therapy: a 4-month, randomized, placebo-controlled trial. AB - AIMS: To evaluate the anti-anginal and anti-ischaemic efficacy of the selective I(f) current inhibitor ivabradine in patients with chronic stable angina pectoris receiving beta-blocker therapy. METHODS AND RESULTS: In this double-blinded trial, 889 patients with stable angina receiving atenolol 50 mg/day were randomized to receive ivabradine 5 mg b.i.d. for 2 months, increased to 7.5 mg b.i.d. for a further 2 months, or placebo. Patients underwent treadmill exercise tests at the trough of drug activity using the standard Bruce protocol for randomization and at 2 and 4 months. Total exercise duration at 4 months increased by 24.3 +/- 65.3 s in the ivabradine group, compared with 7.7 +/- 63.8 s with placebo (P < 0.001). Ivabradine was superior to placebo for all exercise test criteria at 4 months (P < 0.001 for all) and 2 months (P-values between <0.001 and 0.018). Ivabradine in combination with atenolol was well tolerated. Only 1.1% of patients withdrew owing to sinus bradycardia in the ivabradine group. CONCLUSION: The combination of ivabradine 7.5 mg b.i.d. and atenolol at the commonly used dosage in clinical practice in patients with chronic stable angina pectoris produced additional efficacy with no untoward effect on safety or tolerability. PMID- 19136490 TI - Impact of the main implantable cardioverter-defibrillator trials in clinical practice: data from the Italian ICD Registry for the years 2005-07. AB - AIMS: Several studies have demonstrated the life-saving role of implantable cardioverter-defibrillator (ICD) therapy. In order to evaluate the effects of major trials in clinical practice, we report the clinical data of patients enrolled in the Italian ICD Registry for the years 2005-07. METHODS AND RESULTS: The Registry collects prospectively national ICD implantation activity on the basis of EURID Card. The number of ICDs per million of inhabitants was 180.6 in the year 2005, 192.5 in the year 2006, and 220.6 in the year 2007 (P = 0.02). The median age was 69 years for all 3 years considered. Implantable cardioverter defibrillator implantation for primary prevention concerned 4692 patients in 2005 (44.2%), 5938 patients in 2006 (52.2%), 7324 patients in 2007 (55.7%) and for secondary prevention 5921 patients in 2005 (55.8%), 5447 patients in 2006 (47.8%), and 5828 patients in 2007 (44.3%). Single-chamber ICDs were implanted in 30.9, 31.0, and 28.5% of patients, dual chamber ICDs in 31.9, 31.5, and 31.7%, and biventricular ICDs in 37.2, 37.5, and 39.8%, respectively. CONCLUSION: The ICD implantation rate in Italy increased significantly in the period 2005-07. The registry showed a significant increase in prophylactic and biventricular ICD use, reflecting a favourable adherence to trials and guidelines in the clinical practice. PMID- 19136491 TI - Six-minute walking test predicts long-term cardiac death in patients who received cardiac resynchronization therapy. AB - AIMS: Cardiac resynchronization therapy (CRT) has been proven to be effective in patients suffering from chronic heart failure (CHF) associated with electrical dyssynchrony. However, long-term predictors of mortality in that subset have not been extensively investigated. The aim of this study was to establish baseline long-term predictors of cardiovascular mortality in CHF patients treated with CRT. METHODS AND RESULTS: A total of 188 consecutive patients with moderate to severe CHF who had undergone a successful CRT implant were evaluated. Baseline measurements included clinical history, a 6-min walking test (6MWT), and echocardiographic parameters. Patients with cardiac or non-cardiac diseases limiting their ability to perform a 6MWT were excluded, with the final count totalling 155 patients [82% men, mean age 69 +/- 8 years, New York Heart Association (NYHA) functional class: II 22%, III 73.5%, IV 4.5%]. A total of 24 patients (15.5%) died of cardiovascular causes and one patient underwent heart transplantation during a mean follow-up of 24.4 +/- 18.1 months. Univariate analysis showed that NYHA class, distance walked in the 6MWT, left atrial diameter, digoxine and left ventricle (LV) ejection fraction were all significantly related to rates of mortality. Multivariate Cox regression after adjustment for the presence of a defibrillator showed that the LV ejection fraction [HR 0.91 (95% CI: 0.84-0.98) P = 0.008] and 6MWT distance <225 m [HR 5.6 (95% CI: 1.2-25.3) P = 0.026] were independent predictors of cardiovascular mortality. CONCLUSION: Baseline functional capacity, measured by the 6MWT distance, and LV ejection fraction are independent predictors of mortality in moderate to severe CHF patients, despite CRT. A 6MWT distance walked of <225 m identifies patients at high risk of cardiovascular death at mid-long term. PMID- 19136493 TI - Phylogenetic relationships of Cranichidinae and Prescottiinae (Orchidaceae, Cranichideae) inferred from plastid and nuclear DNA sequences. AB - BACKGROUND AND AIMS: Phylogenetic relationships of subtribes Cranichidinae and Prescottiinae, two diverse groups of neotropical terrestrial orchids, are not satisfactorily understood. A previous molecular phylogenetic study supported monophyly for Cranichidinae, but Prescottiinae consisted of two clades not sister to one another. However, that analysis included only 11 species and eight genera of these subtribes. Here, plastid and nuclear DNA sequences are analysed for an enlarged sample of genera and species of Cranichidinae and Prescottiinae with the aim of clarifying their relationships, evaluating the phylogenetic position of the monospecific genera Exalaria, Ocampoa and Pseudocranichis and examining the value of various structural traits as taxonomic markers. METHODS: Approx. 6000 bp of nucleotide sequences from nuclear ribosomal (ITS) and plastid DNA (rbcL, matK trnK and trnL-trnF) were analysed with cladistic parsimony and Bayesian inference for 45 species/14 genera of Cranichidinae and Prescottiinae (plus suitable outgroups). The utility of flower orientation, thickenings of velamen cell walls, hamular viscidium and pseudolabellum to mark clades recovered by the molecular analysis was assessed by tracing these characters on the molecular trees. KEY RESULTS: Spiranthinae, Cranichidinae, paraphyletic Prescottia (with Pseudocranichis embedded), and a group of mainly Andean 'prescottioid' genera (the 'Stenoptera clade') were strongly supported. Relationships among these clades were unresolved by parsimony but the Bayesian tree provided moderately strong support for the resolution (Spiranthinae-(Stenoptera clade (Prescottia/Pseudocranichis-Cranichidinae))). Three of the four structural characters mark clades on the molecular trees, but the possession of a pseudolabellum is variable in the polyphyletic Ponthieva. CONCLUSIONS: No evidence was found for monophyly of Prescottiinae and the reinstatement of Cranichidinae s.l. (including the genera of 'Prescottiinae') is favoured. Cranichidinae s.l. are diagnosed by non-resupinate flowers. Lack of support from parsimony for relationships among the major clades of core spiranthids is suggestive of a rapid morphological radiation or a slow rate of molecular evolution. PMID- 19136492 TI - Clinical and arrhythmic outcomes after implantation of a defibrillator for primary prevention of sudden death in patients with post-myocardial infarction cardiomyopathy: The Survey to Evaluate Arrhythmia Rate in High-risk MI patients (SEARCH-MI). AB - AIMS: To evaluate clinical and arrhythmic outcomes in post-infarction cardiomyopathy patients implanted with a defibrillator (ICD) for primary prevention of sudden death. METHODS AND RESULTS: The SEARCH-MI registry is a European multi-centre, prospective, observational study enrolling patients after myocardial infarction, chronic left ventricular dysfunction and an ICD implanted for primary prevention of sudden death. Data on 556 patients with at least one recorded follow-up are presented. Survey to Evaluate Arrhythmia Rate in High-risk MI (SEARCH-MI) patients were sicker than those enrolled in MADIT-II with higher New York Heart Association class and left bundle branch block. Total mortality was 10.4%. Close to one-third (30%) of patients experienced episodes of sustained ventricular arrhythmia. One-quarter (23%) received at least one appropriate therapy and 10% inappropriate therapy. Gender (25% males vs. 5% females, P = 0.0009) and history of non-sustained ventricular tachycardia (24% with vs. 18% without P = 0.037) were predictive of appropriate ventricular therapy. CONCLUSION: SEARCH-MI represents the current clinical management of post-MI patients with left ventricular dysfunction indicated to defibrillator implant for primary prevention. European routine clinical practice was influenced by landmark trials and guidelines which impacted on the implantation of cardiac resynchronization therapy in over 25% of such patients. Non-sustained ventricular tachycardia identifies subjects with a higher incidence of appropriate ICD therapy. PMID- 19136494 TI - Post-abscission, pre-dispersal seeds of Digitalis purpurea remain in a developmental state that is not terminated by desiccation ex planta. AB - BACKGROUND AND AIMS: Seed quality may be compromised if seeds are harvested before natural dispersal (shedding). It has been shown previously that slow or delayed drying can increase potential quality compared with immediate rapid drying. This study set out to investigate whether or not there is a critical moisture content, below which drying terminates maturation events for seeds harvested after mass maturity but before dispersal. METHODS: Seeds of foxglove (Digitalis purpurea) in the post-abscission pre-dispersal phase were held at between 15 and 95 % RH for 4 or 8 d, with or without re-hydration to 95 % RH for a further 4 d, before drying to equilibrium at 15 % RH. In addition, dry seeds were primed for 48 h at -1 MPa. Subsequent seed longevity was assessed at 60 % RH and 45 degrees C. KEY RESULTS: Rate of germination and longevity were improved by holding seeds at a wide range of humidities after harvest. Longevity was further improved by re-hydration at 95 % RH. Priming improved the longevity of the seeds dried immediately after harvest, but not of those first held at 95 % RH for 8 d prior to drying. CONCLUSIONS: Maturation continued ex planta in these post abscission, pre-dispersal seeds of D. purpurea dried at 15-80 % RH at a rate correlated positively with RH (cf. ageing of mature seeds). Subsequent re hydration at 95 % RH enabled a further improvement in quality. Priming seeds initially stored air-dry for 3 months also allowed maturation events to resume. However, once individual seeds within the population had reached maximum longevity, priming had a negative impact on their subsequent survival. PMID- 19136496 TI - In vitro neurogenesis from neural progenitor cells isolated from the hippocampus region of the brain of adult rats exposed to ethanol during early development through their alcohol-drinking mothers. AB - AIMS: This study was aimed to determine whether ethanol exposure during early development altered neurogenesis in the brain of adult rats. METHODS: Pregnant rats were given either ethanol-mixed or mannose-mixed (for control) rodent liquid diet ad libitum. Ethanol drinking continued during pregnancy and nursing. After weaning, the pups (AC(o): pups from control mothers, AE(o): pups from ethanol exposed mothers) received normal diet and water ad libitum for 11 weeks. Then the rats were anesthetized, their brains were collected and the hippocampal samples were processed for isolation of neural progenitor cells (NPCs). AC(o) NPCs and AE(o) NPCs were sequentially grown in media containing different growth factors that induced proliferation and differentiation. RESULTS AND CONCLUSIONS: Neuronal maturation was significantly delayed in ethanol-exposed rats. AC(o) NPCs, up to day 7 of culture, exhibited high beta-catenin-probe binding, an increase in Ca(2+) when exposed to gamma-amino butyric acid (GABA) and lack of response to glutamate (Glu) exposure. beta-Catenin-probe binding and the stimulatory effects of GABA declined thereafter. AC(o) NPCs, at culture day 29, exhibited high beta catenin-probe binding, lack of response to GABA and elevated Glu-induced increase in Ca(2+i). Cultures of AE(o) NPCs showed an amplified stimulatory effects of GABA, attenuated stimulatory effects of Glu and attenuated the delayed (culture day 29) increase in the expression of Wnt proteins and beta-catenin-probe binding. This suggests a significant alteration in neurogenesis and synapse formation in adult rats exposed to ethanol at early development through their alcohol-drinking mothers. PMID- 19136497 TI - Effect of a small dose of alcohol on the endurance performance of trained cyclists. AB - AIM: The aim of this study was to investigate the effect of an acute small ethanol (EtOH) dose (0.5 ml EtOH/kg fat-free mass, combined with carbohydrate) in a drink on endurance performance of trained cyclists. METHODS: Thirteen well trained male cyclists took part in this study. A 60-min cycling endurance performance test (time trial) was performed in a calorimetric chamber after drinking an EtOH (30 +/- 1.8 ml) or a non-EtOH control (C) drink. RESULTS: Overall, EtOH induced a significant decrease in the average cycling power output (PO) (EtOH: 233 +/- 23 W versus C: 243 +/- 24 W, P < 0.01). The time course of mechanical PO showed an early decrease during the EtOH trial as compared to C (P < 0.01). Due to the lower PO, oxygen consumption, carbon dioxide production and glucose oxidation were significantly lower (P < 0.05) as compared to C. Relative to PO, heart rate response and ratings of perceived exertion (RPE) were increased by EtOH as compared to C (P < 0.05). In contrast, EtOH did not influence gross work efficiency, glycaemia and blood lactate concentration. CONCLUSIONS: These results show that the acute low dose of EtOH decreased endurance performance. An increase of cardio-vascular strain and psychobiological mechanisms may explain this decrease of endurance performance. PMID- 19136498 TI - Proteomics approach in the study of the pathophysiology of alcohol-related brain damage. AB - AIMS: Chronic, excessive drinking of alcohol can induce brain damage in the regions important for neurocognitive function. Some of the damage are permanent while some are appearantly reversible. It is our aim to understand the molecular mechanisms underlying alcohol-induced and/or related brain damage, particularly of that observed in 'medically uncomplicated' (without heptatic cirrhosis or Wernicke-Korsakoff Syndrome [WKS]) alcoholics. METHODS: A high-throughput proteomics technology has been applied to several 'alcohol-sensitive' brain regions from uncomplicated and hepatic cirrhosis-complicated alcoholics to understand the mechanisms of alcohol-related brain damage at the level of protein expression. RESULTS: It was clearly demonstrated that each brain region reacts in significantly different manner to chronic alcohol ingestion. Appearant abnormalities in vitamin B1 (thiamine)-related biochemical pathways were observed in several brain regions, such as the dorsolateral prefrontal cortex, genu (a frontal part of the corpus callosum) and cerebellar vermis in uncomplicated alcoholics, suggesting that the reduction of this important nutritional component might be associated with brain damage even without the signs of WKS. In addition, in the two different subregions of the corpus callosum (genu and splenium [a posterior part of the corpus callosum]) and the cerebellar vermis, significant differences in protein expression profiles between uncomplicated and complicated alcoholics with hepatic cirrhosis were identified, suggesting that hepatic factors such as ammonia have significant additive influences on brain protein expression, which might lead to the structural changes and/or damage in these brain regions. Furthermore, in the hippocampus, significant change of the level of glutamine synthetase expression was observed, suggesting once again the importance of ammonia as a cause of brain damage in this region. CONCLUSIONS: Although our data did not show any evidence of "direct" alcohol effects to induce the alteration of protein expression in association with brain damage, high throughput neuroproteomics approaches are proven to have a potential to dissect the mechanisms of complex brain disorders. PMID- 19136499 TI - Alcohol withdrawal and prolonged hospital stay in a patient with neuroimaging abnormalities: a case report. AB - A hospital stay of 30 days was required in a 47-year-old woman with alcohol withdrawal. Magnetic resonance imaging (MRI) findings revealed a focal brain stem lesion and multiple focal supracortical abnormalities. Could asymptomatic neuroimaging abnormalities predict risk of complicated alcohol withdrawal? Future clinical observations and longitudinal studies may wish to address this potential risk factor. PMID- 19136500 TI - Alterations to cell metabolism in connective tissues of the knee after ovariohysterectomy in a rabbit model: are there implications for the postmenopausal athlete? AB - BACKGROUND: Participation in regular exercise and athletic activities across the lifespan is encouraged to maintain the cardiovascular and musculoskeletal systems and general wellbeing. Before the menopause there is an increased risk of anterior cruciate ligament (ACL) injuries in female athletes, whereas there is an increased risk of joint diseases such as knee osteoarthritis after the menopause. Although there are few data regarding alterations in individual connective tissues of the knee in humans either before, during or after the menopause, it is possible to assess changes in experimental models following surgical menopause. OBJECTIVE: To assess changes in cell metabolism in the medial collateral ligament, ACL, patellar tendon, lateral and medial menisci, tibial plateau and femoral condyle articular cartilage and the synovium after surgical menopause in an experimental model system. METHODS: Panels of rabbits were subjected to ovariohysterectomy or sham operations, and RNA from each tissue was assessed for collagen, proteoglycan, proteinase, growth factor, sex hormone receptor and inflammatory mediator messenger RNA levels by reverse transcribed PCR. RESULTS: Unique alterations in cell metabolism were detected 2 months after surgical menopause and the pattern of significant changes was tissue specific (number of mRNA species altered, extent of changes, elevation/depression of changes). CONCLUSIONS: Changes in cell metabolism may alter the set point for the tissues of the knee and subsequently the functioning of the knee after the menopause. Such changes may contribute to an increased risk of injury and/or degenerative conditions. Further studies in pre and postmenopausal women athletes may also shed light on whether the present findings can be extrapolated to human populations. PMID- 19136501 TI - Creating active environments across the life course: "thinking outside the square". AB - The built environment and physical activity agenda provides a unique opportunity for public health, physical activity and planning researchers to be front and centre of a movement aimed at creating healthier and more sustainable environments. However, in order to optimise environments that encourage physical activity across the life course, researchers in this field need to think beyond their "square" -that is, the target group, setting and physical activity behaviour with which they work. We suggest that researchers working in this field need a better understanding of systems theory to appreciate that a change to one part of a complex system can positively and negatively influence other parts of the system. An understanding of systems theory would help minimise unintended negative consequences to other population subgroups or to other types of physical activity from the implementation of our research findings. In this way, a more comprehensive set of research, practice and programme-related activities may emerge, which will advance physical activity research and practice, and improve population health across the life course. PMID- 19136502 TI - Echocardiographic deformation imaging reveals preserved regional systolic function in endurance athletes with left ventricular hypertrophy. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is often observed in athletes, which should be differentiated from hypertrophic cardiomyopathy. The aim of the study was to explore the functional changes measured using tissue Doppler imaging (TDI) deformation analysis in athletes fulfilling LVH criteria participating in different endurance sports. METHODS: Healthy controls (n = 62, 58% men) and endurance athletes (n = 120, 62% men) aged 18-40 years were prospectively enrolled and underwent both standard echocardiography as well as TDI. Longitudinal TDI-derived strain and strain rate (SR) were calculated in the septal and posterior wall in three segments. LVH was defined as a left ventricular mass (LVM) over 132 g/m2 in men and over 109 g/m2 in women. RESULTS: Echocardiographic LVH was observed in 33 athletes (67% men). LVM was significantly increased in both athlete groups (102.6 g/m2 (SD 16.0) and 135.7 g/m2 (SD 15.9) vs 88.0 g/m2 (SD 16.5) in controls, p<0.001). Diastolic parameters were not significantly different between groups. Athletes with LVH showed no significant difference in strain and SR values in any segment of the septal or posterior wall compared with controls or those without LVH. A weak but significant correlation (also after multivariate analysis) was found for septal wall thickness and LVM in peak systolic strain (r = 0.26, p<0.01 and 0.23, p<0.01) and SR (r = 0.27, p<0.01 and 0.29, p<0.01). Nevertheless, strain and SR values were still within normal limits in all athletes. CONCLUSION: Athletes with LVH overall show normal deformation values in the left ventricle. These data suggest that a moderate reduction in regional septal deformation should not be considered as pathological when evaluating the endurance athlete with echocardiographic LVH of unknown origin. PMID- 19136503 TI - Polymorphisms in the IGF1 signalling pathway including the myostatin gene are associated with left ventricular mass in male athletes. AB - BACKGROUND: Athlete's heart as an adaptation to long-time and intensive endurance training can vary considerably between individuals. Genetic polymorphisms in the cardiological relevant insulin-like growth factor 1 (IGF1) signalling pathway seem to have an essential influence on the extent of physiological hypertrophy. OBJECTIVE: Analysis of polymorphisms in the genes of IGF1, IGF1 receptor (IGF1R) and the negative regulator of the cardiac IGF1 signalling pathway, myostatin (MSTN), and their relation to left ventricular mass (LVM) of endurance athletes. METHODS: In 110 elite endurance athletes or athletes with a high amount of endurance training (75 males and 35 females) and 27 male controls, which were examined by echocardiographic imaging methods and ergometric exercise-testing, the genotypes of a cytosine-adenine repeat polymorphism in the promoter region of the IGF1 gene and a G/A substitution at position 3174 in the IGF1R gene were determined. Additionally, a mutation screen of the MSTN gene was performed. RESULTS: The polymorphisms in the IGF1 and the IGF1R gene showed a significant relation to the LVM for male (IGF1: p=0.003; IGF1R: p=0.01), but not for female athletes. The same applies to a previously unnoticed polymorphism in the 1 intron of the MSTN gene, whose deletion allele (AAA->AA) appears to increase the myostatic effect (p=0.015). Moreover, combinations of the polymorphisms showed significant synergistic effects on the LVM of the male athletes. CONCLUSIONS: The authors' results argue for the importance of polymorphisms in the IGF1 signalling pathway in combination with MSTN on the variant degree of physiological hypertrophy of male athletes. PMID- 19136504 TI - Prophylactic misuse and recommended use of non-steroidal anti-inflammatory drugs by athletes. PMID- 19136505 TI - Fasciitis first before tendinopathy: does the anatomy hold the key? PMID- 19136506 TI - Impact of mode of transportation on dyslipidaemia in working people in Beijing. AB - OBJECTIVE: This study aims to investigate the association between mode of transportation to work and dyslipidaemia. METHODS: During the period between January and February 2006, telephone interviews were conducted with 2506 randomly selected urban residents aged 18 years or older in the 8 districts of Beijing, using a multiple stratified random sampling technique. Of the selected individuals, 1024 (40.86%) members of the workforce were subsequently tested for biomarkers (ie, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C)). Multiple logistic regression modelling was used, adjusted for potential confounders. RESULTS: The probability of dyslipidaemia in workers who travel to work by bus, car or taxi is higher than that of workers who walk to work, with prevalence odds ratios (PORs) of 1.99 (95% CI 1.33 to 2.97) and 2.21 (95% CI 1.28 to 3.84), respectively. There is no significant difference in the risk of experiencing dyslipidaemia when workers who ride bicycles are compared with those who walk to work (POR = 1.22, 95% CI 0.83 to 1.78). CONCLUSIONS: These findings indicate that modes of transportation to work are significantly associated with the prevalence of dyslipidaemia. Prevention education should be emphasised among higher-risk people who usually go to work by car, bus or taxi. PMID- 19136507 TI - Physical inactivity: the biggest public health problem of the 21st century. PMID- 19136508 TI - Monitoring disease activity by stool analyses: from occult blood to molecular markers of intestinal inflammation and damage. AB - It is a common experience that gastrointestinal symptoms urge us to differentiate inflammatory bowel disease (IBD) from functional disorders. Furthermore, in patients with proven IBD the disease activity has to be accurately monitored. Faecal markers of neutrophil influx into the mucosa are promising indicators of intestinal inflammation. Some neutrophil-derived proteins may be linked to the pathogenesis of IBD due to their functions as damage-associated molecular pattern molecules (DAMPs). Phagocyte-specific DAMPs of the S100 family are released from neutrophils or monocytes, followed by pro-inflammatory activation of pattern recognition receptors. The complex of S100A8/S100A9 was termed "calprotectin" and has been in use as a faecal marker for 10 years. More recently, faecal S100A12 has been reported to be an even more accurate faecal marker of inflammation. We review the biology of this novel group of molecules which can be used as surrogate markers directly linked to the molecular mechanisms of gut inflammation. PMID- 19136509 TI - Serum pepsinogens and risk of gastric and oesophageal cancers in the General Population Nutrition Intervention Trial cohort. AB - OBJECTIVE: Low serum pepsinogen I (PGI) and low pepsinogen I/pepsinogen II ratio (PGI/II ratio) are markers of gastric fundic atrophy. We aimed to prospectively test the association between serum PGI/II ratio and risks of gastric non-cardia adenocarcinoma, gastric cardia adenocarcinoma, and oesophageal squamous cell carcinoma (OSCC). DESIGN: Case-cohort study nested in a prospective cohort with over 15 years of follow-up. SETTING: Rural region of the People's Republic of China. SUBJECTS: Men and women aged 40-69 years at study baseline. MAIN OUTCOME MEASURES: Adjusted hazard ratios and 95% confidence intervals for the association between serum PGI/II ratio and cancer risk. RESULTS: Compared to subjects with PGI/II ratio of >4, those with or=8 and the vancomycin Etest was U conversion, the functions and substrate specificities of these gene products vary considerably. For example, although two closely related enzymes, A3F and A3G, both restrict HIV 1 infection in strains deficient in virus infectivity factor (vif), A3F selectively deaminates cytosine within 5'-TTCA-3' motifs in single stranded DNA, whereas A3G targets 5'-CCCA-3' sequences. In the present study we have used nucleoside analog interference mapping to probe A3G-DNA interactions throughout the enzyme-substrate complex as well as to determine which DNA structural features determine substrate specificity. Our results indicate that multiple components of nucleosides within the consensus sequence are important for substrate recognition by A3G (with base moieties being most critical), whereas deamination interference by analog substitution outside this region is minimal. Furthermore, exocyclic groups in pyrimidines 1-2 nucleotides 5' of the target cytosine were shown to dictate substrate recognition by A3G, with chemical composition at ring positions 3 and 4 found to be more important than at ring position 5. Taken together, these results provide insights into how the enzyme selects A3G hotspot motifs for deamination as well as which approaches might be best suited for forming a stable, catalytically competent cross-linked A3G-DNA complex for future structural studies. PMID- 19136563 TI - Poly-L-aspartic acid enhances and prolongs gentamicin-mediated suppression of the CFTR-G542X mutation in a cystic fibrosis mouse model. AB - Aminoglycosides such as gentamicin have the ability to suppress translation termination at premature stop mutations, leading to a partial restoration of protein expression and function. This observation led to studies showing that this approach may provide a viable treatment for patients with genetic diseases such as cystic fibrosis that are caused by premature stop mutations. Although aminoglycoside treatment is sometimes associated with harmful side effects, several studies have shown that the co-administration of polyanions such as poly L-aspartic acid (PAA) can both reduce toxicity and increase the intracellular aminoglycoside concentration. In the current study we examined how the co administration of gentamicin with PAA influenced the readthrough of premature stop codons in cultured cells and a cystic fibrosis mouse model. Using a dual luciferase readthrough reporter system in cultured cells, we found that the co administration of gentamicin with PAA increased readthrough 20-40% relative to cells treated with the same concentration of gentamicin alone. Using a Cftr-/- hCFTR-G542X mouse model, we found that PAA also increased the in vivo nonsense suppression induced by gentamicin. Following the withdrawal of gentamicin, PAA significantly prolonged the time interval during which readthrough could be detected, as shown by short circuit current measurements and immunofluorescence. Because the use of gentamicin to suppress disease-causing nonsense mutations will require their long term administration, the ability of PAA to reduce toxicity and increase both the level and duration of readthrough has important implications for this promising therapeutic approach. PMID- 19136564 TI - Phospholipase Cgamma2 is critical for Dectin-1-mediated Ca2+ flux and cytokine production in dendritic cells. AB - Dectin-1 is a C-type lectin that recognizes beta-glucan in the cell walls of fungi and plays an important role in anti-fungal immunity. It signals via tyrosine kinase Syk and adaptor protein Card9 to activate NF-kappaB leading to proinflammatory cytokine production in dendritic cells (DCs). Other than this, not much else is known of the mechanism of Dectin-1 signaling. We demonstrate here that stimulation of DCs with zymosan triggers an intracellular Ca2+ flux that can be attenuated by a blocking anti-Dectin-1 antibody or by pre-treatment of cells with the phospholipase C (PLC) gamma-inhibitor U73122, suggesting that Dectin-1 signals via a PLCgamma pathway to induce Ca2+ flux in DCs. Interestingly, treatment of DCs with particulate curdlan, which specifically engages Dectin-1, results in the phosphorylation of both PLCgamma1 and PLCgamma2. However, we show that PLCgamma2 is the critical enzyme for Dectin-1 signaling in DCs. PLCgamma2-deficient DCs have drastic impairment of Ca2+ signaling and are defective in their secretion of interleukin 2 (IL-2), IL-6, IL-10, IL-12, IL-23, and tumor necrosis factor alpha. PLCgamma2-deficient DCs also exhibit impaired activation of ERK and JNK MAPKs and AP-1 and NFAT transcription factors in response to Dectin-1 stimulation. In addition, PLCgamma2-deficient DCs are also impaired in their activation of NF-kappaB upon Dectin-1 engagement due to defective assembly of the Card9-Bcl10-Malt1 complex and impaired IKKalpha/beta activation and IkappaBalpha degradation. Thus, our data indicate that pattern recognition receptors such as Dectin-1 could elicit Ca2+ signaling and that PLCgamma2 is a critical player in the Dectin-1 signal transduction pathway. PMID- 19136565 TI - Neuronal differentiation is regulated by leucine-rich acidic nuclear protein (LANP), a member of the inhibitor of histone acetyltransferase complex. AB - Neuronal differentiation is a tightly regulated process characterized by temporal and spatial alterations in gene expression. A number of studies indicate a significant role for histone acetylation in the regulation of genes involved in development. Histone acetylation is regulated by histone deacetylases and histone acetyltransferases. Recent findings suggest that these catalytic activities, in turn, are modulated by yet another set of regulators. Of considerable interest in this context is the possible role of the INHAT (inhibitor of histone acetyltransferase) complex, comprised of a group of acidic proteins that suppress histone acetylation by a novel "histone-masking" mechanism. In this study, we specifically examined the role of the leucine-rich acidic nuclear protein (LANP), a defining member of the INHAT complex whose expression is tightly regulated in neuronal development. We report that depleting LANP in neuronal cell lines promotes neurite outgrowth by inducing changes in gene expression. In addition, we show that LANP directly regulates expression of the neurofilament light chain, an important neuron-specific cytoskeletal gene, by binding to the promoter of this gene and modulating histone acetylation levels. Finally, we corroborated our findings in vivo by demonstrating increased neurite outgrowth in primary neurons obtained from LANP null mice, which is also accompanied by increased histone acetylation at the NF-L promoter. Taken together, these results implicate INHATs as a distinct class of developmental regulators involved in the epigenetic modulation of neuronal differentiation. PMID- 19136566 TI - Plant Vacuolar ATP-binding Cassette Transporters That Translocate Folates and Antifolates in Vitro and Contribute to Antifolate Tolerance in Vivo. AB - The vacuoles of pea (Pisum sativum) leaves and red beet (Beta vulgaris) storage root are major sites for the intracellular compartmentation of folates. In the light of these findings and preliminary experiments indicating that some plant multidrug resistance-associated protein (MRP) subfamily ATP-binding cassette transporters are able to transport compounds of this type, the Arabidopsis thaliana vacuolar MRP, AtMRP1 (AtABCC1), and its functional equivalent(s) in vacuolar membrane vesicles purified from red beet storage root were studied. In so doing, it has been determined that heterologously expressed AtMRP1 and its equivalents in red beet vacuolar membranes are not only competent in the transport of glutathione conjugates but also folate monoglutamates and antifolates as exemplified by pteroyl-l-glutamic acid and methotrexate (MTX), respectively. In agreement with the results of these in vitro transport measurements, analyses of atmrp1 T-DNA insertion mutants of Arabidopsis ecotypes Wassilewskia and Columbia disclose an MTX-hypersensitive phenotype. atmrp1 knock out mutants are more sensitive than wild-type plants to growth retardation by nanomolar concentrations of MTX, and this is associated with impaired vacuolar antifolate sequestration. The vacuoles of protoplasts isolated from the leaves of Wassilewskia atmrp1 mutants accumulate 50% less [(3)H]MTX than the vacuoles of protoplasts from wild-type plants when incubated in media containing nanomolar concentrations of this antifolate, and vacuolar membrane-enriched vesicles purified from the mutant catalyze MgATP-dependent [(3)H]MTX uptake at only 40% of the capacity of the equivalent membrane fraction from wild-type plants. AtMRP1 and its counterparts in other plant species therefore have the potential for participating in the vacuolar accumulation of folates and related compounds. PMID- 19136567 TI - Letter to the editor: Comment on 'Acute impact of immediate release methylphenidate administered three times a day on sleep in children with attention-deficit/hyperactivity disorder'. PMID- 19136568 TI - Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphigidae) and its natural host Nicotiana attenuata. VIII. An unbiased GCxGC-ToFMS analysis of the plant's elicited volatile emissions. AB - Treating wounds in Nicotiana attenuata leaves with Manduca sexta oral secretions (W+OS) mimics most changes elicited by M. sexta herbivory, but an unbiased analysis of the effect of the different OS constituents on volatile emissions is lacking. We used two-dimensional gas chromatography/time-of-flight (GCxGC-ToF) mass spectrometry combined with multivariate statistics to parse volatiles into regulatory patterns. Volatiles released by wounding alone and by the alkalinity of OS were assessed by applying a buffer known to mimic the pH-mediated changes of OS elicitation (pectin methyl esterase activation and methanol release). The activities of fatty acid amino acid conjugates, well-known elicitors of antiherbivore defenses, and of 2-hydroxyoctadecatrienoic acid, a newly discovered signal in OS, were determined. Approximately 400 analytes were detected after deconvolution and alignment of GCxGC data; 35 volatiles were significantly regulated upon W+OS. Two-thirds of these were specifically regulated by OS, being either amplified (most terpenoids and certain hexenylesters) or strongly repressed (many short-chain alcohols and some aromatic and hexenylester derivatives). Fatty acid amino acid conjugates played a central role in this pattern of regulation, since they induced the emission of half of OS-elicited volatiles and inhibited the production of almost all OS-repressed volatiles; 2 hydroxyoctadecatrienoic acid influenced emission of trans-alpha-bergamotene, while other unknown OS constituents amplified hexenylester production. We conclude that the complex bouquet of herbivory-elicited volatiles results from the complex modulations of the wound response by diverse cues found in OS. This work also underscores the value of ultra-high-resolution GCxGC-ToF analysis combined with the nontargeted mining of the resulting data. PMID- 19136569 TI - Tyrosine and phenylalanine are synthesized within the plastids in Arabidopsis. AB - While the presence of a complete shikimate pathway within plant plastids is definitively established, the existence of a cytosolic postchorismate portion of the pathway is still debated. This question is alimented by the presence of a chorismate mutase (CM) within the cytosol. Until now, the only known destiny of prephenate, the product of CM, is incorporation into tyrosine (Tyr) and/or phenylalanine (Phe). Therefore, the presence of a cytosolic CM suggests that enzymes involved downstream of CM in Tyr or Phe biosynthesis could be present within the cytosol of plant cells. It was thus of particular interest to clarify the subcellular localization of arogenate dehydrogenases (TYRAs) and arogenate dehydratases (ADTs), which catalyze the ultimate steps in Tyr and Phe biosynthesis, respectively. The aim of this study was to address this question in Arabidopsis (Arabidopsis thaliana) by analysis of the subcellular localization of the two TYRAAts and the six AtADTs. This article excludes the occurrence of a spliced TYRAAt1 transcript encoding a cytosolic TYRA protein. Transient expression analyses of TYRA- and ADT-green fluorescent protein fusions reveal that the two Arabidopsis TYRA proteins and the six ADT proteins are all targeted within the plastid. Accordingly, TYRA and ADT proteins were both immunodetected in the chloroplast soluble protein fraction (stroma) of Arabidopsis. No TYRA or ADT proteins were immunodetected in the cytosol of Arabidopsis cells. Taken together, all our data exclude the possibility of Tyr and/or Phe synthesis within the cytosol, at least in green leaves and Arabidopsis cultured cells. PMID- 19136570 TI - Effects of FSH on testicular mRNA transcript levels in the hypogonadal mouse. AB - FSH acts through the Sertoli cell to ensure normal testicular development and function. To identify transcriptional mechanisms through which FSH acts in the testis, we have treated gonadotrophin-deficient hypogonadal (hpg) mice with recombinant FSH and measured changes in testicular transcript levels using microarrays and real-time PCR 12, 24 and 72 h after the start of treatment. Approximately 400 transcripts were significantly altered at each time point by FSH treatment. At 12 h, there was a clear increase in the levels of a number of known Sertoli cell transcripts (e.g. Fabp5, Lgals1, Tesc, Scara5, Aqp5). Additionally, levels of Leydig cell transcripts were also markedly increased (e.g. Ren1, Cyp17a1, Akr1b7, Star, Nr4a1). This was associated with a small but significant rise in testosterone at 24 and 72 h. At 24 h, androgen-dependent Sertoli cell transcripts were up-regulated (e.g. Rhox5, Drd4, Spinlw1, Tubb3 and Tsx) and this trend continued up to 72 h. By contrast with the somatic cells, only five germ cell transcripts (Dkkl1, Hdc, Pou5f1, Zfp541 and 1700021K02Rik) were altered by FSH within the time-course of the experiment. Analysis of canonical pathways showed that FSH induced a general decline in transcripts related to formation and regulation of tight junctions. Results show that FSH acts directly and indirectly to induce rapid changes in Sertoli cell and Leydig cell transcript levels in the hpg mouse but that effects on germ cell development must occur over a longer time-span. PMID- 19136571 TI - The sequence after the signal peptide of the G protein-coupled endothelin B receptor is required for efficient translocon gating at the endoplasmic reticulum membrane. AB - The heptahelical G protein-coupled receptors (GPCRs) must reach their correct subcellular location to exert their function. Receptor domains relevant for receptor trafficking include signal sequences mediating receptor integration into the membrane of the endoplasmic reticulum (ER) and anterograde or retrograde transport signals promoting receptor sorting into the vesicles of the secretory pathway. In addition, receptors must be correctly folded to pass the quality control system of the early secretory pathway. Taking the endothelin B receptor as a model, we describe a new type of a transport-relevant GPCR domain. Deletion of this domain (residues Glu(28) to Trp(54)) leads to a fully functional receptor protein that is expressed at a lower level than the wild-type receptor. Subcellular localization experiments and glycosylation state analyses demonstrate that the mutant receptor is neither misfolded, retained intracellularly, nor misrouted. Fluorescence recovery after photobleaching analyses demonstrate that constitutive internalization is also not affected. By using an in vitro prion protein targeting assay, we show that this domain is necessary for efficient translocon gating at the ER membrane during early receptor biogenesis. Taken together, we identified a novel transport-relevant domain in the GPCR protein family. Our data may also be relevant for other GPCRs and unrelated integral membrane proteins. PMID- 19136572 TI - The G-alpha protein GNA3 of Hypocrea jecorina (Anamorph Trichoderma reesei) regulates cellulase gene expression in the presence of light. AB - Although the enzymes enabling Hypocrea jecorina (anamorph Trichoderma reesei) to degrade the insoluble substrate cellulose have been investigated in some detail, little is still known about the mechanism by which cellulose signals its presence to the fungus. In order to investigate the possible role of a G-protein/cyclic AMP signaling pathway, the gene encoding GNA3, which belongs to the adenylate cyclase-activating class III of G-alpha subunits, was cloned. gna3 is clustered in tandem with the mitogen-activated protein kinase gene tmk3 and the glycogen phosphorylase gene gph1. The gna3 transcript is upregulated in the presence of light and is almost absent in the dark. A strain bearing a constitutively activated version of GNA3 (gna3QL) exhibits strongly increased cellulase transcription in the presence of the inducer cellulose and in the presence of light, whereas a gna3 antisense strain showed delayed cellulase transcription under this condition. However, the gna3QL mutant strain was unable to form cellulases in the absence of cellulose. The necessity of light for stimulation of cellulase transcription by GNA3 could not be overcome in a mutant which expressed gna3 under control of the constitutive gpd1 promoter also in darkness. We conclude that the previously reported stimulation of cellulase gene transcription by light, but not the direct transmission of the cellulose signal, involves the function and activation of GNA3. The upregulation of gna3 by light is influenced by the light modulator ENVOY, but GNA3 itself has no effect on transcription of the light regulator genes blr1, blr2, and env1. Our data for the first time imply an involvement of a G-alpha subunit in a light-dependent signaling event in fungi. PMID- 19136573 TI - Disruption of the Aopex11-1 gene involved in peroxisome proliferation leads to impaired Woronin body formation in Aspergillus oryzae. AB - The Woronin body, a unique organelle found in the Pezizomycotina, plugs the septal pore upon hyphal damage to prevent excessive cytoplasmic bleeding. Although it was previously shown that the Woronin body buds out from the peroxisome, the relationship between peroxisomal proliferation/division and Woronin body differentiation has not been extensively investigated. In this report, we examined whether Pex11 required for peroxisomal proliferation participates in Woronin body formation in Aspergillus oryzae. A. oryzae contained two orthologous PEX11 genes that were designated Aopex11-1 and Aopex11-2. Deletion of Aopex11 genes revealed that only the DeltaAopex11-1 strain showed reduced growth and enlarged peroxisomes in the presence of oleic acid as a sole carbon source, indicating a defect in peroxisomal function and proliferation. Disruption of Aopex11-1 gene impaired the Woronin body function, leading to excessive loss of the cytosol upon hyphal injury. Dual localization analysis of the peroxisome and Woronin body protein AoHex1 demonstrated that Woronin bodies fail to fully differentiate from peroxisomes in the DeltaAopex11-1 strain. Furthermore, distribution of AoHex1 was found to be peripheral in the enlarged peroxisome or junctional in dumbbell-shaped peroxisomes. Electron microscopy of the DeltaAopex11-1 strain revealed the presence of Woronin bodies that remained associated with organelles resembling peroxisomes, which was supported from the sucrose gradient centrifugation confirming that the Woronin body protein AoHex1 overlapped with the density-shifted peroxisome in the DeltaAopex11-1 strain. In conclusion, the present study describes the role of Pex11 in Woronin body differentiation for the first time. PMID- 19136574 TI - Hypoxia-induced mitogenic factor (HIMF/FIZZ1/RELMalpha) induces the vascular and hemodynamic changes of pulmonary hypertension. AB - Pulmonary hypertension (PH) is a serious disease of multiple etiologies mediated by hypoxia, immune stimuli, and elevated pulmonary pressure that leads to vascular thickening and eventual right heart failure. In a chronic hypoxia model of PH, we previously reported the induction of a novel pleiotropic cytokine, hypoxia-induced mitogenic factor (HIMF), that exhibits mitogenic, vasculogenic, contractile, and chemokine properties during PH-associated vascular remodeling. To examine the role of HIMF in hypoxia-induced vascular remodeling, we performed in vivo knockdown of HIMF using short hairpin RNA directed at rat HIMF in the chronic hypoxia model of PH. Knockdown of HIMF partially blocked increases in mean pulmonary artery pressure, pulmonary vascular resistance, right heart hypertrophy, and vascular remodeling caused by chronic hypoxia. To demonstrate a direct role for HIMF in the mechanism of PH development, we performed HIMF-gene transfer into the lungs of rats using a HIMF-expressing adeno-associated virus (AAV). AAV-HIMF alone caused development of PH similar to that of chronic hypoxia with increased mean pulmonary artery pressure and pulmonary vascular resistance, right heart hypertrophy, and neomuscularization and thickening of small pulmonary arterioles. The findings suggest that HIMF represents a critical cytokine-like growth factor in the development of PH. PMID- 19136575 TI - IL-1beta-induced cortisol stimulates lung fluid absorption in fetal guinea pigs via SGK-mediated Nedd4-2 inhibition. AB - We tested the hypothesis that interleukin (IL)-1beta-induced cortisol synthesis stimulates distal lung fluid absorption in fetal guinea pigs via induction of serum- and glucocorticoid-regulated kinase (SGK) and inhibition of neural precursor cell expressed, developmentally downregulated protein 4-2 (Nedd4-2). IL 1beta was subcutaneously administered daily to timed-pregnant guinea pigs over 3 days. Fetuses were obtained by abdominal hysterotomy at gestation day (GD)61 and GD68 and instilled with an isosmolar 5% albumin solution into the lungs. Distal lung fluid movement was measured over 1 h from the change in distal air space protein concentration. Fetal lungs were secreting lung fluid at GD61 while absorbing lung fluid at GD68. Distal lung fluid absorption was induced at GD61 by IL-1beta but unaffected at GD68. Plasma cortisol concentrations were increased by IL-1beta at GD61 and endogenously at GD68. Distal lung fluid absorption was measured and correlated to SGK and Nedd4-2 expression and to alpha-epithelial Na channel (ENaC) expression. SGK was increased by IL-1beta and late during gestation (GD68), while Nedd4-2 was decreased by IL-1beta and late during gestation. alpha-ENaC was induced by IL-1beta at GD61 and increased late during gestation. Thus our study suggests that cortisol-stimulated fetal lung fluid absorption is mediated by increased ENaC expression and may be governed by the SGK/Nedd4-2 pathway. These observations may explain why babies delivered preterm after intrauterine inflammation have a reduced risk of developing severe respiratory distress. PMID- 19136576 TI - Neutrophil elastase cleaves VEGF to generate a VEGF fragment with altered activity. AB - Excessive neutrophil elastase (NE) activity and altered vascular endothelial growth factor (VEGF) signaling have independently been implicated in the development and progression of pulmonary emphysema. In the present study, we investigated the potential link between NE and VEGF. We noted that VEGF(165) is a substrate for NE. Digestion of purified VEGF(165) with NE generated a partially degraded disulfide-linked fragment of VEGF. Mass spectrometric analysis revealed that NE likely cleaves VEGF(165) at both the NH(2) and COOH termini to produce VEGF fragment chains approximately 5 kDa reduced in size. NE treatment of VEGF laden endothelial cell cultures and smooth muscle cells endogenously expressing VEGF generated VEGF fragments similar to those observed with purified VEGF(165). NE-generated VEGF fragment showed significantly reduced binding to VEGF receptor 2 and heparin yet retained the ability to bind to VEGF receptor 1. Interestingly, VEGF fragment showed altered signaling in pulmonary artery endothelial cells compared with intact VEGF(165). Specifically, treatment with VEGF fragment did not activate extracellular-regulated kinases 1 and 2 (ERK1/2), yet resulted in enhanced activation of protein kinase B (Akt). Treatment of monocyte/macrophage RAW 264.7 cells with VEGF fragment, on the other hand, led to both Akt and ERK1/2 activation, increased VEGFR1 expression, and stimulated chemotaxis. These findings suggest that the tissue response to NE-mediated injury might involve the generation of diffusible VEGF fragments that stimulate inflammatory cell recruitment and activation via VEGF receptor 1. PMID- 19136577 TI - Mechanisms of eosinophil major basic protein-induced hyperexcitability of vagal pulmonary chemosensitive neurons. AB - We have reported recently that eosinophil-derived basic proteins directly enhance the capsaicin- and electrical stimulation-evoked whole cell responses in rat pulmonary sensory neurons (19). Our present study further elucidates the mechanisms underlying the sensitization of pulmonary afferent nerves induced by these cationic proteins. Our results show that pretreatment with eosinophil major basic protein (MBP; 2 microM, 60 s) significantly enhanced the excitability of isolated rat vagal pulmonary chemosensitive neurons to acid and ATP in the current-clamp mode, but this potentiating effect was absent in the voltage-clamp recordings. The hyperexcitability induced by MBP was not prevented by the blockade of either transient receptor potential vanilloid type-1 receptor (TRPV1) selectively (inhibitor: AMG 9810; 1 microM, 2 min) or all TRPV1-4 channels (inhibitor: ruthenium red; 5 microM, 2 min). In addition, MBP also markedly potentiated the excitability of mouse pulmonary chemosensitive neurons, and no detectable difference was found between those isolated from wild-type and TRPV1 knockout mice. Furthermore, MBP pretreatment affected the decay time and recovery phase of the action potentials evoked by current injections and significantly inhibited both the sustained delayed-rectifier voltage-gated K(+) current (IK(dr)) and the A-type, fast-inactivating K(+) current (IK(a)) in these sensory neurons. In conclusion, our results indicate that the inhibition of IK(dr) and IK(a) should, at least in part, account for the hyperexcitability of pulmonary chemosensitive neurons induced by eosinophil-derived cationic proteins, whereas an interaction with TRPV1 channels does not seem to be required for the sensitizing effect of these proteins. PMID- 19136578 TI - Carbon dioxide inhalation causes pulmonary inflammation. AB - The aim of this study was to assess whether one of the most common poisons of cellular respiration, i.e., carbon dioxide, is proinflammatory. CO(2) is naturally present in the atmosphere at the level of 0.038% and involved in numerous cellular biochemical reactions. We analyzed in vitro the inflammation response induced by exposure to CO(2) for 48 h (0-20% with a constant O(2) concentration of 21%). In vivo mice were submitted to increasing concentrations of CO(2) (0, 5, 10, and 15% with a constant O(2) concentration of 21%) for 1 h. The exposure to concentrations above 5% of CO(2) resulted in the increased transcription (RNase protection assay) and secretion (ELISA) of proinflammatory cytokines [macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, MIP-2, IL-8, IL-6, monocyte chemoattractant protein-1, and regulated upon activation, normal T cell expressed, and, presumably, secreted (RANTES)] by epithelial cell lines HT-29 or A549 and primary pulmonary cells retrieved from the exposed mice. Lung inflammation was also demonstrated in vivo by mucin 5AC-enhanced production and airway hyperreactivity induction. This response was mostly mediated by the nuclear translocation of p65 NF-kappaB, itself a consequence of protein phosphatase 2A (PP2A) activation. Short inhibiting RNAs (siRNAs) targeted toward PP2Ac reversed the effect of carbon dioxide, i.e., disrupted the NF-kappaB activation and the proinflammatory cytokine secretion. In conclusion, this study strongly suggests that exposure to carbon dioxide may be more toxic than previously thought. This may be relevant for carcinogenic effects of combustion products. PMID- 19136579 TI - Role of clathrin-mediated endocytosis of surfactant protein A by alveolar macrophages in intracellular signaling. AB - We recently provided evidence that anti-inflammatory macrophage activation, i.e., the inhibition of constitutive and signal-induced NF-kappaB activity by the pulmonary collectin surfactant protein (SP)-A, critically involves a promoted stabilization of IkappaB-alpha, the predominant inhibitor of NF-kappaB, via posttranscriptional mechanisms comprising the activation of atypical (a)PKCzeta. SP-A uptake and degradation by alveolar macrophages (AMphi) occur in a receptor mediated, clathrin-dependent manner. However, a mutual link between endocytosis of and signaling by SP-A remains elusive. The aim of this study was to investigate whether clathrin-mediated endocytosis (CME) of SP-A by AMphi is a prerequisite for its modulation of the IkappaB-alpha/NF-kappaB pathway. The inhibition of clathrin-coated pit (CCP) formation and clathrin-coated vesicle (CCV) formation/budding abrogates SP-A-mediated IkappaB-alpha stabilization and SP-A-mediated inhibition of LPS-induced NF-kappaB activation in freshly isolated rat AMphi, as determined by Western analysis, fluorescence-activated cell sorting, confocal microscopy, and EMSA. Actin depolymerization and inhibition of CCP formation further abolished SP-A-mediated inhibition of LPS-induced TNF-alpha release, as determined by ELISA. In addition, SP-A-induced atypical PKCzeta activation was abolished by pretreatment of AMphi with CCV inhibitors as determined by in vitro immunocomplex kinase assay. Although CME is classically considered as a means to terminate signaling, our results demonstrate that SP-A uptake via CME by AMphi has to precede the initiation of SP-A signaling. PMID- 19136580 TI - Heat shock protein 90-eNOS interactions mature with postnatal age in the pulmonary circulation of the piglet. AB - Binding of endothelial nitric oxide synthase (eNOS) to the chaperone protein, Hsp90, promotes coupled eNOS synthetic activity. Using resistance level pulmonary arteries (PRA) from 2-day-, 5- to 7-day-, and 12-day-old piglets, we tested the hypothesis that Hsp90-eNOS interactions are developmentally regulated in the early neonatal period. PRA were isolated for coimmunoprecipitation and immunoblot analyses or cannulated for continuous diameter measurements using the pressurized myography technique. NOS inhibition caused less constriction in PRA from 2-day- compared with 5- to 7-day- and 12-day-old piglets. No age-related differences were found in dilation responses to an NO donor or in protein expression of Hsp90, phospho-eNOS (Ser(1177)), Akt, phospho-Akt, or caveolin-1. Compared with the older animals, PRA from 2-day-old piglets had higher total eNOS expression but displayed less binding of eNOS to Hsp90 and Akt. Hsp90 antagonism with radicicol induced greatest constriction in PRA from 12-day-old piglets. ACh stimulation caused dilation in PRA from 5- to 7-day- and 12-day-old but not 2-day old animals, despite rapid and equivalent ACh-mediated eNOS phosphorylation (Ser(1177)) in all three age groups. Hsp90 inhibition abolished ACh-mediated dilation in PRA from the older piglets. ACh failed to stimulate Hsp90-eNOS binding in 2-day-old but induced a significant increase in Hsp90-eNOS coimmunoprecipitation in PRA from the older age groups, which was blocked by Hsp90 antagonism. We conclude that physical interactions between Hsp90 and eNOS mature over the first weeks of life, likely contributing to the postnatal fall in pulmonary vascular resistance and changes in agonist-induced pulmonary vascular responses characteristic of the early neonatal period. PMID- 19136581 TI - Variable stretch pattern enhances surfactant secretion in alveolar type II cells in culture. AB - Secretion of pulmonary surfactant that maintains low surface tension within the lung is primarily mediated by mechanical stretching of alveolar epithelial type II (AEII) cells. We have shown that guinea pigs ventilated with random variations in frequency and tidal volume had significantly larger pools of surfactant in the lung than animals ventilated in a monotonous manner. Here, we test the hypothesis that variable stretch patterns imparted on the AEII cells results in enhanced surfactant secretion. AEII cells isolated from rat lungs were exposed to equibiaxial strains of 12.5, 25, or 50% change in surface area (DeltaSA) at 3 cycles/min for 15, 30, or 60 min. (3)H-labeled phosphatidylcholine release and cell viability were measured 60 min following the onset of stretch. Whereas secretion increased following 15-min stretch at 50% DeltaSA and 30-min stretch at 12.5% DeltaSA, 60 min of cyclic stretch diminished surfactant secretion regardless of strain. When cells were stretched using a variable strain profile in which the amplitude of each stretch was randomly pulled from a uniform distribution, surfactant secretion was enhanced both at 25 and 50% mean DeltaSA with no additional cell injury. Furthermore, at 50% mean DeltaSA, there was an optimum level of variability that maximized secretion implying that mechanotransduction in these cells exhibits a phenomenon similar to stochastic resonance. These results suggest that application of variable stretch may enhance surfactant secretion, possibly reducing the risk of ventilator-induced lung injury. Variable stretch-induced mechanotransduction may also have implications for other areas of mechanobiology. PMID- 19136582 TI - Long-term effects of prenatal hypoxia on endothelium-dependent relaxation responses in pulmonary arteries of adult sheep. AB - Chronic hypoxia during the course of pregnancy is a common insult to the fetus. However, its long-term effect on the pulmonary vasculature in adulthood has not been described. In this study, the vasorelaxation responses of conduit pulmonary arteries in adult female sheep that were chronically hypoxic as fetuses and raised postnatally at sea level were investigated. Vessel tension studies revealed that endothelium-dependent relaxation responses were attenuated in pulmonary arteries from adult sheep that experienced prenatal hypoxia. Endothelial nitric oxide synthase (eNOS) protein expression was unchanged, but eNOS activity was significantly decreased in pulmonary arteries from prenatally hypoxic sheep. Protein expression of eNOS partners, caveolin-1, calmodulin, and heat shock protein 90 (Hsp90) did not change following prenatal hypoxia. However, the association between eNOS and caveolin-1, its inhibitory binding partner, was significantly increased, whereas association between eNOS and its stimulatory partners calmodulin and Hsp90 was greatly decreased. Furthermore, phosphorylation of Ser(1177) in eNOS decreased, whereas phosphorylation of Thr(495) increased, in the prenatally hypoxic pulmonary arteries, events that are related to eNOS activity. These data demonstrate that prenatal hypoxia results in persistent abnormalities in endothelium-dependent relaxation responses of pulmonary arteries in adult sheep due to decreased eNOS activity resulting from altered posttranslational regulation. PMID- 19136583 TI - Surface expression of CD74 by type II alveolar epithelial cells: a potential mechanism for macrophage migration inhibitory factor-induced epithelial repair. AB - Macrophage migration inhibitory factor (MIF) is a pleiotropic proinflammatory cytokine involved in acute lung injury and other processes such as wound repair and tumor growth. MIF exerts pro-proliferative effects on a variety of cell types including monocytes/macrophages, B cells, and gastric epithelial cell lines through binding to the major histocompatibility complex type II-associated invariant chain, CD74. In acute lung injury, inflammatory damage of the alveolar epithelium leads to loss of type I alveolar epithelial cells (AEC-I), which are replaced by proliferation and differentiation of type II alveolar epithelial cells (AEC-II). In this study we have investigated the potential of MIF to contribute to alveolar repair by stimulating alveolar epithelial cell proliferation. We show that murine AEC-II, but not AEC-I, express high surface levels of CD74 in vivo. Culture of AEC-II in vitro resulted in decreased mRNA levels for CD74 and loss of surface CD74 expression, which correlated with a transition of AEC-II to an AEC-I-like phenotype. MIF stimulation of AEC-II induced rapid and prolonged phosphorylation of ERK1/2 and Akt, increased expression of cyclins D1 and E, as well as AEC-II proliferation. Corresponding MIF signaling and enhanced thymidine incorporation was observed after MIF stimulation of MLE-12 cells transfected to overexpress CD74. In contrast, MIF did not induce MAPK activation, gene transcription, or increased proliferation in differentiated AEC-I-like cells that lack CD74. These data suggest a previously unidentified role of MIF-CD74 interaction by inducing proliferation of AEC-II, which may contribute to alveolar repair. PMID- 19136584 TI - Glutamine in acute lung injury: the experimental model matters. PMID- 19136585 TI - Enhanced expression of beta 3-galactosyltransferase 5 activity is sufficient to induce in vivo synthesis of extended type 1 chains on lactosylceramides of selected human colonic carcinoma cell lines. AB - In general, an elevated expression of beta 3-galactosyltransferase (beta 3GalT) activity contributed by beta 3GalT5 correlates well with increased biosynthesis and expression of type 1 chain (Gal beta 1-3GlcNAc beta 1-) derivatives such as Lewis A and sialyl Lewis A, which are mostly recognized as terminal epitopes and not further extended. Most known beta 3-N-acetylglucosaminyltransferases show a higher activity toward extending type 2 chain (Gal beta 1-4GlcNAc beta 1-), and an over-expression of beta 3GalT5 could suppress the formation of the type 2 chain poly-N-acetyllactosaminoglycans. The potential of extending instead the predominant type 1 chain termini synthesized under such circumstances was, however, not investigated, partly due to technical difficulty in unambiguous identification of extended type 1 chains. Using an advanced mass spectrometry based glycomic mapping and glycan sequencing approach, we show here that type 1 chains carried on the lacto-series glycosphingolipids of colonic carcinoma cells can be extended when the endogenous beta 3GalT activity relative to competing beta 4GalT activity, as defined against a common GlcNAc beta 1-3Gal beta 1-4Glc acceptor, is sufficiently high, as found in Colo205 and SW1116, but not in DLD-1 cells. In support of this positive correlation, the lacto-series glycosphingolipids isolated from stably transfected DLD-1 clones over-expressing beta 3GalT5 were shown to comprise fucosylated dimeric type 1 chains, whereas a mock transfectant and the DLD-1 parent carried only fucosylated dimeric type 2 chains on their lactosylceramides. It suggests that while the natural expression of extended type 1 chain is likely to be determined by many contributing factors including the relative amounts of competing glycosyltransferases and the UDP-Gal level, the enhanced expression of beta 3GalT5 is sufficient to promote in vivo extension of type 1 chains by furnishing a significantly higher amount of type 1 chain precursors relative to competing type 2 chains. PMID- 19136586 TI - Autogenous regulation of Escherichia coli polynucleotide phosphorylase expression revisited. AB - The Escherichia coli polynucleotide phosphorylase (PNPase; encoded by pnp), a phosphorolytic exoribonuclease, posttranscriptionally regulates its own expression at the level of mRNA stability and translation. Its primary transcript is very efficiently processed by RNase III, an endonuclease that makes a staggered double-strand cleavage about in the middle of a long stem-loop in the 5'-untranslated region. The processed pnp mRNA is then rapidly degraded in a PNPase-dependent manner. Two non-mutually exclusive models have been proposed to explain PNPase autogenous regulation. The earlier one suggested that PNPase impedes translation of the RNase III-processed pnp mRNA, thus exposing the transcript to degradative pathways. More recently, this has been replaced by the current model, which maintains that PNPase would simply degrade the promoter proximal small RNA generated by the RNase III endonucleolytic cleavage, thus destroying the double-stranded structure at the 5' end that otherwise stabilizes the pnp mRNA. In our opinion, however, the first model was not completely ruled out. Moreover, the RNA decay pathway acting upon the pnp mRNA after disruption of the 5' double-stranded structure remained to be determined. Here we provide additional support to the current model and show that the RNase III-processed pnp mRNA devoid of the double-stranded structure at its 5' end is not translatable and is degraded by RNase E in a PNPase-independent manner. Thus, the role of PNPase in autoregulation is simply to remove, in concert with RNase III, the 5' fragment of the cleaved structure that both allows translation and prevents the RNase E-mediated PNPase-independent degradation of the pnp transcript. PMID- 19136587 TI - Involvement of the Cra global regulatory protein in the expression of the iscRSUA operon, revealed during studies of tricarballylate catabolism in Salmonella enterica. AB - In Salmonella enterica, tricarballylate (Tcb) catabolism requires function of TcuB, a membrane-bound protein that contains [4Fe-4S] clusters and heme. TcuB transfers electrons from reduced flavin adenine dinucleotide in the Tcb dehydrogenase (TcuA) to electron acceptors in the membrane. We recently showed that functions needed to assemble [Fe-S] clusters (i.e., the iscRSUA-hscBA-fdx operon) compensate for the lack of ApbC during growth of an apbC strain on Tcb. ApbC had been linked to [Fe-S] cluster metabolism, and we showed that an apbC strain had decreased TcuB activity. Here we report findings that expand our understanding of the regulation of expression of the iscRSUA genes in Salmonella enterica. We investigated why low levels of glucose or other saccharides restored growth of an apbC strain on Tcb. Here we report the following findings. (i) A < or =1 mM concentration of glucose, fructose, ribose, or glycerol restores growth of an apbC strain on Tcb. (ii) The saccharide effect results in increased levels of TcuB activity. (iii) The saccharide effect depends on the global regulatory protein Cra. (iv) Putative Cra binding sites are present in the regulatory region of the iscRSUA operon. (v) Cra protein binds to all three sites in the iscRSUA promoter region in a concentration-dependent fashion. To our knowledge, this is the first report of the involvement of Cra in [Fe-S] cluster assembly. PMID- 19136589 TI - Analysis by mutagenesis of a chromosomal integron integrase from Shewanella amazonensis SB2BT. AB - Integrons are mobile genetic elements that can integrate and disseminate genes as cassettes by a site-specific recombination mechanism. Integrons contain an integrase gene (intI) that carries out recombination by interacting with two different target sites; the attI site in cis with the integrase and the palindromic attC site of a cassette. The plasmid-specified IntI1 excises a greater variety of cassettes (principally antibiotic resistance genes), and has greater activity, than chromosomal integrases. The aim of this study was to analyze the capacity of the chromosomal integron integrase SamIntIA of the environmental bacterium Shewanella amazonensis SB2BT to excise various cassettes and to compare the properties of the wild type with those of mutants that substitute consensus residues of active integron integrases. We show that the SamIntIA integrase is very weakly active in the excision of various cassettes but that the V206R, V206K, and V206H substitutions increase its efficiency for the excision of cassettes. Our results also suggest that the cysteine residue in the beta-5 strand is essential to the activity of Shewanella-type integrases, while the cysteine in the beta-4 strand is less important for the excision activity. PMID- 19136588 TI - Flagellar biogenesis of Xanthomonas campestris requires the alternative sigma factors RpoN2 and FliA and is temporally regulated by FlhA, FlhB, and FlgM. AB - In prokaryotes, flagellar biogenesis is a complicated process involving over 40 genes. The phytopathogen Xanthomonas campestris pv. campestris possesses a single polar flagellum, which is essential for the swimming motility. A sigma54 activator, FleQ, has been shown to be required for the transcriptional activation of the flagellar type III secretion system (F-T3SS), rod, and hook proteins. One of the two rpoN genes, rpoN2, encoding sigma54, is essential for flagellation. RpoN2 and FleQ direct the expression of a second alternative sigma FliA (sigma28) that is essential for the expression of the flagellin FliC. FlgM interacts with FliA and represses the FliA regulons. An flgM mutant overexpressing FliC generates a deformed flagellum and displays an abnormal motility. Mutation in the two structural genes of F-T3SS, flhA and flhB, suppresses the production of FliC. Furthermore, FliA protein levels are decreased in an flhB mutant. A mutant defective in flhA, but not flhB, exhibits a decreased infection rate. In conclusion, the flagellar biogenesis of Xanthomonas campestris requires alternative sigma factors RpoN2 and FliA and is temporally regulated by FlhA, FlhB, and FlgM. PMID- 19136590 TI - ClpX inhibits FtsZ assembly in a manner that does not require its ATP hydrolysis dependent chaperone activity. AB - ClpX is a well-characterized bacterial chaperone that plays a role in many processes, including protein turnover and the remodeling of macromolecular complexes. All of these activities require ATP hydrolysis-dependent, ClpX mediated protein unfolding. Here we used site-directed mutagenesis in combination with genetics and biochemistry to establish that ClpX inhibits assembly of the conserved division protein FtsZ through a noncanonical mechanism independent of its role as an ATP-dependent chaperone. PMID- 19136591 TI - The Rcs two-component system regulates expression of lysozyme inhibitors and is induced by exposure to lysozyme. AB - The Escherichia coli Rcs regulon is triggered by antibiotic-mediated peptidoglycan stress and encodes two lysozyme inhibitors, Ivy and MliC. We report activation of this pathway by lysozyme and increased lysozyme sensitivity when Rcs induction is genetically blocked. This lysozyme sensitivity could be alleviated by complementation with Ivy and MliC. PMID- 19136592 TI - In Bacillus subtilis, the sirtuin protein deacetylase, encoded by the srtN gene (formerly yhdZ), and functions encoded by the acuABC genes control the activity of acetyl coenzyme A synthetase. AB - This report provides in vivo evidence for the posttranslational control of the acetyl coenzyme A (Ac-CoA) synthetase (AcsA) enzyme of Bacillus subtilis by the acuA and acuC gene products. In addition, both in vivo and in vitro data presented support the conclusion that the yhdZ gene of B. subtilis encodes a NAD(+)-dependent protein deacetylase homologous to the yeast Sir2 protein (also known as sirtuin). On the basis of this new information, a change in gene nomenclature, from yhdZ to srtN (for sirtuin), is proposed to reflect the activity associated with the YdhZ protein. In vivo control of B. subtilis AcsA function required the combined activities of AcuC and SrtN. Inactivation of acuC or srtN resulted in slower growth and cell yield under low-acetate conditions than those of the wild-type strain, and the acuC srtN strain grew under low acetate conditions as poorly as the acsA strain. Our interpretation of the latter result was that both deacetylases (AcuC and SrtN) are needed to maintain AcsA as active (i.e., deacetylated) so the cell can grow with low concentrations of acetate. Growth of an acuA acuC srtN strain on acetate was improved over that of the acuA(+) acuC srtN strain, indicating that the AcuA acetyltransferase enzyme modifies (i.e., inactivates) AcsA in vivo, a result consistent with previously reported in vitro evidence that AcsA is a substrate of AcuA. PMID- 19136593 TI - The atypical hybrid histidine protein kinase RodK in Myxococcus xanthus: spatial proximity supersedes kinetic preference in phosphotransfer reactions. AB - Many proteins of two-component signal transduction systems (TCS) have domain structures that do not comply with a phosphate flow as observed in linear TCS, phosphorelays, or simple branched pathways. An example is RodK, which is essential for fruiting body formation in Myxococcus xanthus and, in addition to a sensor domain, consists of a kinase domain and three receiver domains (RodK-R1, R2, and -R3), all of which are functionally important. We identified the RokA response regulator as part of the RodK pathway. In vitro the isolated RodK kinase domain engages in phosphotransfer to RodK-R3 and RokA, with a kinetic preference for RokA. However, in the context of the full-length protein, the RodK kinase domain has a preference for phosphotransfer to RodK-R3 over RokA. We suggest that in full-length RodK, the spatial proximity of the RodK kinase domain and RodK-R3 compensate for the kinetic preference of the isolated kinase domain for RokA. Thus, the kinetic preference observed using an isolated kinase domain of a hybrid kinase does not necessarily reflect the phosphotransfer preference of the full length protein. We speculate that the phosphorylation status of RodK-R1 and RodK R2 determines whether RodK engages in phosphotransfer to RodK-R3 or RokA in vivo. PMID- 19136594 TI - Isolation and characterization of superdormant spores of Bacillus species. AB - Superdormant spores of Bacillus subtilis and Bacillus megaterium were isolated in 4 to 12% yields following germination with high nutrient levels that activated one or two germinant receptors. These superdormant spores did not germinate with the initial nutrients or those that stimulated other germinant receptors, and the superdormant spores' defect was not genetic. The superdormant spores did, however, germinate with Ca(2+)-dipicolinic acid or dodecylamine. Although these superdormant spores did not germinate with high levels of nutrients that activated one or two nutrient germinant receptors, they germinated with nutrient mixtures that activated more receptors, and using high levels of nutrient mixtures activating more germinant receptors decreased superdormant spore yields. The use of moderate nutrient levels to isolate superdormant spores increased their yields; the resultant spores germinated poorly with the initial moderate nutrient concentrations, but they germinated well with high nutrient concentrations. These findings suggest that the levels of superdormant spores in populations depend on the germination conditions used, with fewer superdormant spores isolated when better germination conditions are used. These findings further suggest that superdormant spores require an increased signal for triggering spore germination compared to most spores in populations. One factor determining whether a spore is superdormant is its level of germinant receptors, since spore populations with higher levels of germinant receptors yielded lower levels of superdormant spores. A second important factor may be heat activation of spore populations, since yields of superdormant spores from non-heat-activated spore populations were higher than those from optimally activated spores. PMID- 19136595 TI - Covalently linked AcrB giant offers a new powerful tool for mechanistic analysis of multidrug efflux in bacteria. PMID- 19136596 TI - Heterologous expression of mycobacterial proteins in Saccharomyces cerevisiae reveals two physiologically functional 3-hydroxyacyl-thioester dehydratases, HtdX and HtdY, in addition to HadABC and HtdZ. AB - We report on Mycobacterium tuberculosis Rv0241c and Rv3389c, representing two physiologically functional 3-hydroxyacyl-thioester dehydratases (Htd). These enzymes are potentially entrained in type 2 fatty acid synthase (FASII). Mycobacterial FASII is involved in the synthesis of mycolic acids, which are the major constituents of the protective layer around the pathogen, shielding it from noxious chemicals and the host's immune system. Mycolic acids are additionally associated with the virulence and resilience of M. tuberculosis. Here, Rv0241c and Rv3389c, which are distinct from the previously identified heterodimers Rv0635-Rv0636 (HadAB) and Rv0636-Rv0637 (HadBC) but also the homodimer Rv0130 (HtdZ), were identified by expressing the corresponding candidate open reading frames in Saccharomyces cerevisiae htd2Delta cells lacking mitochondrial 3 hydroxyacyl-acyl carrier protein dehydratase activity, followed by scoring for phenotype rescue. The htd2Delta mutant fails to produce sufficient levels of lipoic acid and does not respire or grow on nonfermentable carbon sources. Soluble protein extracts made from mutant htd2Delta cells expressing mitochondrially targeted Rv0241c or Rv3389c contained 3-hydroxyacyl-thioester hydratase activity. Moreover, mutant yeast cells expressing Rv0241c or Rv3389c were able to recover their respiratory growth on glycerol medium and efficiently reduce 2,3,5-triphenyltetrazolium chloride. Additionally, expression of mitochondrial Rv0241c or Rv3389c in htd2Delta cells also restored de novo lipoic acid synthesis to 92 and 40% of the level in the wild-type strain, respectively. We propose naming Rv0241c and Rv3389c as HtdX and HtdY, respectively, and discuss the implications of our finding with reference to Rv0098, a candidate mycobacterial FabZ homologue with intrinsic thioesterase and hydratase activities that lacks the eukaryotic-like hydratase-2 motif. PMID- 19136597 TI - African 1, an epidemiologically important clonal complex of Mycobacterium bovis dominant in Mali, Nigeria, Cameroon, and Chad. AB - We have identified a clonal complex of Mycobacterium bovis present at high frequency in cattle in population samples from several sub-Saharan west-central African countries. This closely related group of bacteria is defined by a specific chromosomal deletion (RDAf1) and can be identified by the absence of spacer 30 in the standard spoligotype typing scheme. We have named this group of strains the African 1 (Af1) clonal complex and have defined the spoligotype signature of this clonal complex as being the same as the M. bovis BCG vaccine strain but with the deletion of spacer 30. Strains of the Af1 clonal complex were found at high frequency in population samples of M. bovis from cattle in Mali, Cameroon, Nigeria, and Chad, and using a combination of variable-number tandem repeat typing and spoligotyping, we show that the population of M. bovis in each of these countries is distinct, suggesting that the recent mixing of strains between countries is not common in this area of Africa. Strains with the Af1 specific deletion (RDAf1) were not identified in M. bovis isolates from Algeria, Burundi, Ethiopia, Madagascar, Mozambique, South Africa, Tanzania, and Uganda. Furthermore, the spoligotype signature of the Af1 clonal complex has not been identified in population samples of bovine tuberculosis from Europe, Iran, and South America. These observations suggest that the Af1 clonal complex is geographically localized, albeit to several African countries, and we suggest that the dominance of the clonal complex in this region is the result of an original introduction into cows naive to bovine tuberculosis. PMID- 19136598 TI - Functional analysis of insertion sequence ISAba1, responsible for genomic plasticity of Acinetobacter baumannii. AB - ISAba1 is an insertion sequence that is widely distributed in Acinetobacter baumannii. We demonstrated here that ISAba1 and the composite transposon Tn2006 are capable of transposition, generating 9-bp target site duplications. The expression of the ISAba1 transposase-encoding gene was downregulated by translational frameshifting. PMID- 19136599 TI - Complete and draft genome sequences of six members of the Aquificales. AB - The Aquificales are widespread in marine and terrestrial hydrothermal environments. Here, we report the complete and draft genome sequences of six new members of the Aquificales: two marine species, Persephonella marina strain EX-H1 and Hydrogenivirga strain 128-5-R1 (from the East Pacific Rise, 9 degrees 50.3'N, 104 degrees 17.5'W, and the Eastern Lau Spreading Center, 176 degrees 11.5'W, 20 degrees 45.8'S, respectively), and four terrestrial isolates, Sulfurihydrogenibium azorense strain Az-Fu1, Sulfurihydrogenibium yellowstonense strain SS-5, and Sulfurihydrogenibium strain Y03AOP1 (from Furnas, Azores, Portugal, and Calcite Springs and Obsidian Pool in Yellowstone National Park, United States, respectively), and the only thermoacidophilic isolate, Hydrogenobaculum strain Y04AAS1 (from a stream adjacent to Obsidian Pool). Significant differences among the different species exist that include nitrogen metabolism, hydrogen utilization, chemotaxis, and signal transduction, providing insights into their ecological niche adaptations. PMID- 19136600 TI - Synergism of coupled subsarcolemmal Ca2+ clocks and sarcolemmal voltage clocks confers robust and flexible pacemaker function in a novel pacemaker cell model. AB - Recent experimental studies have demonstrated that sinoatrial node cells (SANC) generate spontaneous, rhythmic, local subsarcolemmal Ca(2+) releases (Ca(2+) clock), which occur during late diastolic depolarization (DD) and interact with the classic sarcolemmal voltage oscillator (membrane clock) by activating Na(+) Ca(2+) exchanger current (I(NCX)). This and other interactions between clocks, however, are not captured by existing essentially membrane-delimited cardiac pacemaker cell numerical models. Using wide-scale parametric analysis of classic formulations of membrane clock and Ca(2+) cycling, we have constructed and initially explored a prototype rabbit SANC model featuring both clocks. Our coupled oscillator system exhibits greater robustness and flexibility than membrane clock operating alone. Rhythmic spontaneous Ca(2+) releases of sarcoplasmic reticulum (SR)-based Ca(2+) clock ignite rhythmic action potentials via late DD I(NCX) over much broader ranges of membrane clock parameters [e.g., L type Ca(2+) current (I(CaL)) and/or hyperpolarization-activated ("funny") current (I(f)) conductances]. The system Ca(2+) clock includes SR and sarcolemmal Ca(2+) fluxes, which optimize cell Ca(2+) balance to increase amplitudes of both SR Ca(2+) release and late DD I(NCX) as SR Ca(2+) pumping rate increases, resulting in a broad pacemaker rate modulation (1.8-4.6 Hz). In contrast, the rate modulation range via membrane clock parameters is substantially smaller when Ca(2+) clock is unchanged or lacking. When Ca(2+) clock is disabled, the system parametric space for fail-safe SANC operation considerably shrinks: without rhythmic late DD I(NCX) ignition signals membrane clock substantially slows, becomes dysrhythmic, or halts. In conclusion, the Ca(2+) clock is a new critical dimension in SANC function. A synergism of the coupled function of Ca(2+) and membrane clocks confers fail-safe SANC operation at greatly varying rates. PMID- 19136603 TI - Respiratory modulation of heart sound morphology. AB - Heart sounds, the acoustic vibrations produced by the mechanical processes of the cardiac cycle, are modulated by respiratory activity. We have used computational techniques of cluster analysis and classification to study the effects of the respiratory phase and the respiratory resistive load on the temporal and morphological properties of the first (S1) and second heart sounds (S2), acquired from 12 healthy volunteers. Heart sounds exhibited strong morphological variability during normal respiration and nearly no variability during apnea. The variability was shown to be periodic, with its estimated period in good agreement with the measured duration of the respiratory cycle. Significant differences were observed between properties of S1 and S2 occurring during inspiration and expiration. S1 was commonly attenuated and slightly delayed during inspiration, whereas S2 was accentuated and its aortic component occurred earlier at late inspiration and early expiration. Typical split morphology was observed for S1 and S2 during inspiration. At high-breathing load, these changes became more prominent and occurred earlier in the respiratory cycle. Unsupervised cluster analysis was able to automatically identify the distinct morphologies associated with different respiratory phases and load. Classification of the respiration phase (inspiration or expiration) from the morphology of S1 achieved an average accuracy of 87 +/- 7%, and classification of the breathing load was accurate in 82 +/- 7%. These results suggest that quantitative heart sound analysis can shed light on the relation between respiration and cardiovascular mechanics and may be applied to continuous cardiopulmonary monitoring. PMID- 19136601 TI - 20-HETE increases survival and decreases apoptosis in pulmonary arteries and pulmonary artery endothelial cells. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE) is an endogenous cytochrome P-450 product present in vascular smooth muscle and uniquely located in the vascular endothelium of pulmonary arteries (PAs). 20-HETE enhances reactive oxygen species (ROS) production of bovine PA endothelial cells (BPAECs) in an NADPH oxidase dependent manner and is postulated to promote angiogenesis via activation of this pathway in systemic vascular beds. We tested the capacity of 20-HETE or a stable analog of this compound, 20-hydroxy-eicosa-5(Z),14(Z)-dienoic acid, to enhance survival and protect against apoptosis in BPAECs stressed with serum starvation. 20-HETE produced a concentration-dependent increase in numbers of starved BPAECs and increased 5-bromo-2'-deoxyuridine incorporation. Caspase-3 activity, nuclear fragmentation studies, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays supported protection from apoptosis and enhanced survival of starved BPAECs treated with a single application of 20-HETE. Protection from apoptosis depended on intact NADPH oxidase, phosphatidylinositol 3 (PI3)-kinase, and ROS production. 20-HETE-stimulated ROS generation by BPAECs was blocked by inhibition of PI3-kinase or Akt activity. These data suggest 20-HETE-associated protection from apoptosis in BPAECs required activation of PI3-kinase and Akt and generation of ROS. 20-HETE also protected against apoptosis in BPAECs stressed by lipopolysaccharide, and in mouse PAs exposed to hypoxia reoxygenation ex vivo. In summary, 20-HETE may afford a survival advantage to BPAECs through activation of prosurvival PI3-kinase and Akt pathways, NADPH oxidase activation, and NADPH oxidase-derived superoxide. PMID- 19136602 TI - Temporal changes in expression of connexin 43 after load-induced hypertrophy in vitro. AB - Upon remodeling of the ventricle after a provoking stimulus, such as hypertension, connections between adjacent myocytes may need to be "reformatted" to preserve a synchronization of excitation of the remodeling heart. In the mammalian heart, the protein connexin forms the gap junctions that allow electrical and chemical signaling communication between neighboring cells. We aim to elucidate whether mechanical load, in isolation, potentially changes the expression of connexin 43 (Cx43), the major isoform of the connexin family in the ventricle, and its phosphorylation. Cx43 expression levels and contractile function of multicellular rabbit cardiac preparations were assessed in a newly developed in vitro system that allows for the study of the transition of healthy multicellular rabbit myocardium to hypertrophied myocardium. We found that in mechanically loaded cardiac trabeculae, Cx43 levels remained stable for about 12 h and then rapidly declined. Phosphorylation at Ser368 declined much faster, being almost absent after 2 h of high-load conditions. No-load conditions did not affect Cx43 levels, nor did phosphorylation at Ser368. The downregulation of Cx43 under mechanical load did not correspond with the contractile changes that were observed. Furthermore, blocking paracrine activity of the muscle could only partially prevent the downregulation of Cx43. Additionally, no effect of mechanical loading on the expression of N-cadherin and zonula occludens-1 was observed, indicating a specificity of the connexin response. High mechanical load induced a rapid loss of Cx43 phosphorylation, followed by a decrease in Cx43 protein levels. Paracrine factors are partly responsible for the underlying mechanism of action, whereas no direct correlation to contractile ability was observed. PMID- 19136604 TI - Cardiomyocytes from postinfarction failing rat hearts have improved ischemia tolerance. AB - Altered myocardial Ca(2+) and Na(+) handling in congestive heart failure (CHF) may be expected to decrease the tolerance to ischemia by augmenting reperfusion Ca(2+) overload. The aim of the present study was to investigate tolerance to hypoxia-reoxygenation by measuring enzyme release, cell death, ATP level, and cell Ca(2+) and Na(+) in cardiomyocytes from failing rat hearts. CHF was induced in Wistar rats by ligation of the left coronary artery during isoflurane anesthesia, after which cardiac failure developed within 6 wk. Isolated cardiomyocytes were cultured for 24 h and subsequently exposed to 4 h of hypoxia and 2 h of reoxygenation. Cell damage was measured as lactate dehydrogenase (LD) release, cell death as propidium iodide uptake, and ATP by firefly luciferase assay. Cell Ca(2+) and Na(+) were determined with radioactive isotopes, and free intracellular Ca(2+) concentration ([Ca(2+)](i)) with fluo-3 AM. CHF cells showed less increase in LD release and cell death after hypoxia-reoxygenation and had less relative reduction in ATP level after hypoxia than sham cells. CHF cells accumulated less Na(+) than sham cells during hypoxia (117 vs. 267 nmol/mg protein). CHF cells maintained much lower [Ca(2+)](i) than sham cells during hypoxia (423 vs. 1,766 arbitrary units at 4 h of hypoxia), and exchangeable Ca(2+) increased much less in CHF than in sham cells (1.4 vs. 6.7 nmol/mg protein) after 120 min of reoxygenation. Ranolazine, an inhibitor of late Na(+) current, significantly attenuated both the increase in exchangeable Ca(2+) and the increase in LD release in sham cells after reoxygenation. This supports the suggestion that differences in Na(+) accumulation during hypoxia cause the observed differences in Ca(2+) accumulation during reoxygenation. Tolerance to hypoxia and reoxygenation was surprisingly higher in CHF than in sham cardiomyocytes, probably explained by lower hypoxia-mediated Na(+) accumulation and subsequent lower Ca(2+) accumulation in CHF after reoxygenation. PMID- 19136606 TI - The hexosamine biosynthesis inhibitor azaserine prevents endothelial inflammation and dysfunction under hyperglycemic condition through antioxidant effects. AB - Hexosamine biosynthetic pathway (HBP) accounts for some cardiovascular adverse effects of hyperglycemia. We investigated whether the HBP inhibitor azaserine protects against hyperglycemia-induced endothelial damage dependently of HBP. Human endothelial cells isolated from umbilical veins were exposed either to a high (30.5 mmol/l) or low concentration of glucose (5.5 mmol/l) for 4 days, followed by a stimulation with TNF-alpha (1 ng/ml, 24 h). The blockade of the rate-limiting enzyme glutamine:fructose-6-phosphate amidotransferase inhibited HBP flux and oxidative stress (generation of superoxide and peroxynitrite) under the hyperglycemic condition and prevented the synergistic stimulation of VCAM-1 and ICAM-1 expression by hyperglycemia and TNF-alpha. In the cells cultured under a low-glucose condition when no increased HBP flux occurred, azaserine enhanced the manganese-superoxide dismutase (MnSOD) protein level and also inhibited the oxidative stress and the expression of VCAM-1 and ICAM-1 in response to TNF alpha. Moreover, the polyphenol resveratrol inhibited the oxidative stress and adhesion molecule expression and did not decrease the HBP flux under the hyperglycemia condition. In addition, in isolated rat aortas exposed to hyperglycemic buffer for 5 h when no significant HBP flux occurred, azaserine upregulated the MnSOD protein level and prevented decreased endothelium-dependent relaxations to acetylcholine. In conclusion, hyperglycemia independently increases oxidative stress and HBP flux, amplifies endothelial inflammation, and impairs endothelial function mainly through oxidative stress and not the HBP pathway. Azaserine protects against hyperglycemic endothelial damage through its antioxidant effect independently of inhibiting HBP pathway. PMID- 19136605 TI - Granulocyte colony-stimulating factor protects cardiac mitochondria in the early phase of cardiac injury. AB - Although granulocyte colony-stimulating factor (G-CSF) reportedly plays a cardioprotective role in several models of cardiac injury, clinical use of this drug in cardiac patients has been controversial. Here, we tested, in vivo and in vitro, the effect of G-CSF on cardiac mitochondria, which play a key role in determining cardiac cellular fate and function. Mild stimulation of C57/BL6 mice with doxorubicin (Dox) did not induce cardiac apoptosis or fibrosis but did induce damage to mitochondrial organization of the myocardium as observed through an electron microscope. Cardiac catheterization and echocardiography revealed that Dox did not alter cardiac systolic function or left ventricular size but did reduce diastolic function, an early sign of cardiac damage. Treatment with G-CSF attenuated significantly the damage to mitochondrial organization and rescued diastolic function. In an in vitro model for rat neonatal cardiomyocytes, a subapoptotic dose of Dox induced severe mitochondrial damage, including marked swelling of the cardiac mitochondria and/or decreased mitochondrial membrane potential. These mitochondrial changes were completely blocked by pretreatment with G-CSF. In addition, G-CSF dramatically improved ATP generation, which rescued Dox-impaired mitochondrial electron transport and oxygen consumption mainly through complex IV. These findings clearly indicate that G-CSF protects cardiac mitochondria, which are key organelles in the determination of cardiac cellular fate, in the early phase of cardiac injury. PMID- 19136607 TI - Interatrial shunt for chronic pulmonary hypertension: differential impact of low flow vs. high-flow shunting. AB - The purpose of the present study was to determine for the first time the qualitative and quantitative impact of varying degrees of interatrial shunting on right heart dynamics and systemic perfusion in subjects with chronic pulmonary hypertension (CPH). Eight dogs underwent 3 mo of progressive pulmonary artery banding, following which right atrial and ventricular end-systolic and end diastolic pressure-volume relations were calculated using conductance catheters. An 8-mm shunt prosthesis was inserted between the superior vena cava and left atrium, yielding a controlled model of atrial septostomy. Data were obtained 1) preshunt or "CPH"; 2) "Low-Flow" shunt; and 3) "High-Flow" shunt (occluding superior vena cava forcing all flow through the shunt). With progressive shunting, right ventricular pressure fell from 72 +/- 19 mmHg (CPH) to 54 +/- 17 mmHg (Low-Flow) and 47 +/- 17 mmHg (High-Flow) (P < 0.001). Cardiac output increased from 1.5 +/- 0.3 l/min at CPH to 1.8 +/- 0.4 l/min at Low-Flow (286 +/- 105 ml/min, 15% of cardiac output; P < 0.001), but returned to 1.6 +/- 0.3 l/min at High-Flow (466 +/- 172 ml/min, 29% of cardiac output; P = 0.008 vs. Low-Flow, P = 0.21 vs. CPH). There was a modest rise in systemic oxygen delivery from 252 +/- 46 ml/min at CPH to 276 +/- 50 ml/min at Low-Flow (P = 0.07), but substantial fall to 222 +/- 50 ml/min at High-Flow (P = 0.005 vs. CPH, P < 0.001 vs. Low Flow). With progressive shunting, bichamber contractility did not change (P = 0.98), but the slope of the right atrial end-diastolic pressure volume relation decreased (P < 0.04), consistent with improved compliance. This study demonstrated that Low-Flow interatrial shunting consistently improved right atrial mechanics and systemic perfusion in subjects with CPH, while High-Flow exceeded an "ideal shunt fraction". PMID- 19136608 TI - Adaptive response of the heart to long-term anemia induced by iron deficiency. AB - Anemia is common in patients with chronic heart failure and an independent predictor of poor prognosis. Chronic anemia leads to left ventricular (LV) hypertrophy and heart failure, but its molecular mechanisms remain largely unknown. We investigated the mechanisms, including the molecular signaling pathway, of cardiac remodeling induced by iron deficiency anemia (IDA). Weanling Sprague-Dawley rats were fed an iron-deficient diet for 20 wk to induce IDA, and the molecular mechanisms of cardiac remodeling were evaluated. The iron-deficient diet initially induced severe anemia, which resulted in LV hypertrophy and dilation with preserved systolic function associated with increased serum erythropoietin (Epo) concentration. Cardiac STAT3 phosphorylation and VEGF gene expression increased by 12 wk of IDA, causing angiogenesis in the heart. Thereafter, sustained IDA induced upregulation of cardiac hypoxia inducible factor-1alpha gene expression and maintained upregulation of cardiac VEGF gene expression and cardiac angiogenesis; however, sustained IDA promoted cardiac fibrosis and lung congestion, with decreased serum Epo concentration and cardiac STAT3 phosphorylation after 20 wk of IDA compared with 12 wk. Upregulation of serum Epo concentration and cardiac STAT3 phosphorylation is associated with a beneficial adaptive mechanism of anemia-induced cardiac hypertrophy, and later decreased levels of these molecules may be critical for the transition from adaptive cardiac hypertrophy to cardiac dysfunction in long-term anemia. Understanding the mechanism of cardiac maladaptation to anemia may lead to a new strategy for treatment of chronic heart failure with anemia. PMID- 19136610 TI - If parents establish a no-smoking agreement with their offspring, does this prevent adolescents from smoking? Findings from three Dutch studies. AB - Data from three studies were used to investigate whether the establishment of a no-smoking agreement is related to lower odds of adolescent smoking. The prevalence of a no-smoking agreement was first explored by using a national sample involving 4,501 Dutch adolescents. Second, data from a longitudinal study among 595 early adolescents and their parents were used to test whether establishing a no-smoking agreement prevents adolescents from smoking. Third, the authors tested among 856 early- and mid-adolescents and their parents, whether in addition to the establishment of a no-smoking agreement, the frequency and quality of communication on smoking issues had an effect on adolescent smoking. The findings do not support that establishing a no-smoking agreement is an effective deterrent with regard to adolescent smoking. Parents who want to prevent smoking might consider focusing their efforts on establishing a good quality of communication on smoking issues whereas parents who just talk a lot about smoking issues without considering the quality of their communication might do more harm than good. PMID- 19136609 TI - Cardiac fibroblasts require focal adhesion kinase for normal proliferation and migration. AB - Migration and proliferation of cardiac fibroblasts (CFs) play an important role in the myocardial remodeling process. While many factors have been identified that regulate CF growth and migration, less is known about the signaling mechanisms involved in these processes. Here, we utilized Cre-LoxP technology to obtain focal adhesion kinase (FAK)-deficient adult mouse CFs and studied how FAK functioned in modulating cell adhesion, proliferation, and migration of these cells. Treatment of FAK(flox/flox) CFs with Ad/Cre virus caused over 70% reduction of FAK protein levels within a cell population. FAK-deficient CFs showed no changes in focal adhesions, cell morphology, or protein expression levels of vinculin, talin, or paxillin; proline-rich tyrosine kinase 2 (Pyk2) expression and activity were increased. Knockdown of FAK protein in CFs increased PDGF-BB-induced proliferation, while it reduced PDGF-BB-induced migration. Adhesion to fibronectin was not altered. To distinguish between the function of FAK and Pyk2, FAK function was inhibited via adenoviral-mediated overexpression of the natural FAK inhibitor FAK-related nonkinase (FRNK). Ad/FRNK had no effect on Pyk2 expression, inhibited the PDGF-BB-induced migration, but did not change the PDGF-BB-induced proliferation. FAK deficiency had only modest effects on increasing PDGF-BB activation of p38 and JNK MAPKs, with no alteration in the ERK response vs. control cells. These results demonstrate that FAK is required for the PDGF-BB-induced migratory response of adult mouse CFs and suggest that FAK could play an essential role in the wound-healing response that occurs in numerous cardiac pathologies. PMID- 19136611 TI - Perspectives of mothers in farmworker households on reducing the take-home pathway of pesticide exposure. AB - Farmworkers carry pesticide residue home on their clothing, boots, and skin, placing other household members at risk, particularly children. Specific precautions are recommended to reduce this take-home pathway, yet few studies have examined the perspectives of farmworkers and other household members regarding these behaviors and the reasons for or against adoption. The authors conducted semistructured interviews with 37 Mexican/Mexican-American women in farmworker households to explore the family and cultural context in which pesticide safety practices are performed and to identify factors that facilitate or hinder their adoption. Whereas women could describe the take-home pathway, they were less able to connect it with their family's susceptibility to pesticide exposure. Women experienced difficulty integrating the prevention behaviors into their everyday lives because of competing responsibilities, conflicts with their husbands' intentions and with cultural health beliefs, perceived lack of control, and community barriers that interfered with women's motivations. Implications for practice are discussed. PMID- 19136612 TI - Transcriptional profiling reveals a critical role for tyrosine phosphatase VE-PTP in regulation of VEGFR2 activity and endothelial cell morphogenesis. AB - To define molecular events accompanying formation of the 3-dimensional (3D) vascular tube, we have characterized gene expression during vascular endothelial growth factor (VEGF)-induced tubular morphogenesis of endothelial cells. Microarray analyses were performed comparing gene induction in growth-arrested, tube-forming endothelial cells harvested from 3D collagen cultures to that in proliferating endothelial cells cultured on fibronectin. Differentially expressed genes were clustered and analyzed for specific endothelial expression through publicly available datasets. We validated the contribution of one of the identified genes, vascular endothelial protein tyrosine phosphatase (VE-PTP), to endothelial morphogenesis. Silencing of VE-PTP expression was accompanied by increased VEGF receptor-2 (VEGFR2) tyrosine phosphorylation and activation of downstream signaling pathways. The increased VEGFR2 activity promoted endothelial cell cycle progression, overcoming the G(0)/G(1) arrest associated with organization into tubular structures in the 3D cultures. Proximity ligation showed close association between VEGFR2 and VE-PTP in resting cells. Activation of VEGFR2 by VEGF led to rapid loss of association, which was resumed with time in parallel with decreased receptor activity. In conclusion, we have identified genes, which may serve critical functions in formation of the vascular tube. One of these, VE-PTP, regulates VEGFR2 activity thereby modulating the VEGF-response during angiogenesis. PMID- 19136613 TI - Down-regulation of the hepatic selenoprotein biosynthesis machinery impairs selenium metabolism during the acute phase response in mice. AB - The acute-phase response (APR) is characterized by an impaired metabolism of the essential trace element selenium (Se). Moreover, low-Se concentrations correlate to mortality risk in sepsis. Therefore, we analyzed the expression of the central Se transport and storage protein selenoprotein P (Sepp1) during an APR in mice. Serum Se and Sepp1 concentrations declined in parallel after injection of lipopolysaccharide to 50 and 39% of control-injected littermates, respectively. This negative APR proceeded largely independent from hepatic Sepp1 transcript concentrations. Instead, we identified a set of hepatic transcripts involved in Se metabolism, which declined coordinately during the APR, including the selenocysteine-specific elongation factor (EFsec), selenophosphate-synthetase 2 (Sephs2), selenocysteine-tRNA[Ser]Sec synthase (SecS), and phosphoseryl tRNA[Ser]Sec kinase (Pstk). Pstk reacted most strongly and qualified as a new limiting factor for Sepp1 biosynthesis in siRNA-mediated knockdown experiments in hepatocytes in culture. Analogous experiments were performed with mice transgenic for hepatocyte-specific human Sepp1 cDNA to verify this hypothesis. Similar kinetics and effect sizes of Sepp1 expression were observed as before in wild type mice. We conclude that hepatic translation of Sepp1 mRNA is specifically impaired during the APR. This deficit disrupts regular Se metabolism, transport, and supply to peripheral tissues and likely aggravates the pathological status. PMID- 19136614 TI - Monomeric C-reactive protein activates endothelial cells via interaction with lipid raft microdomains. AB - Emerging evidence indicates that in addition to native pentameric C-reactive protein (CRP), monomeric CRP (mCRP) also plays an active role in inflammation associated with cardiovascular diseases. mCRP activates endothelial cells, one of the critical events in cardiovascular diseases; however, the underlying molecular mechanisms are incompletely understood. Here we report that association of mCRP with human aortic and coronary artery endothelial cells is predominantly due to membrane insertion rather than binding to the surface proteins Fc gammaRs and proteoglycans. We identify lipid rafts as the preferential membrane microdomains for mCRP anchorage. mCRP binding depends on membrane cholesterol content and is synergistically mediated by the putative cholesterol binding consensus sequence of CRP (aa 35-47) and the C-terminal octapeptide (aa 199-206). Conversely, disrupting lipid rafts with methyl-beta cyclodextrin or nystatin abrogated mCRP induced cytokine release, reactive oxygen species generation, and adhesion molecule expression in endothelial cells. Furthermore, ex vivo treatment of rabbit thoracic aorta and carotid artery segments with nystatin prevented mCRP induced IL-8 release. Our data identify mCRP-lipid raft interaction as an important mechanism in mediating cellular responses to mCRP and lend further support to the notion of mCRP regulation of endothelial cell function during inflammation. PMID- 19136615 TI - Differential inflammatory activity across human abdominal aortic aneurysms reveals neutrophil-derived leukotriene B4 as a major chemotactic factor released from the intraluminal thrombus. AB - Development and progression of acquired abdominal aortic aneurysms (AAAs) have been associated with different inflammatory mediators. The aim of the present study was to elucidate the topology and the potential mechanisms linking the leukotriene pathway to human AAAs. Human aneurysmal lesions were obtained from 24 patients undergoing surgery, and the intraluminal thrombus was separated from the vascular wall. Histological examination revealed major expression of the leukotriene-producing enzymes 5-lipoxygenase and LTA(4) hydrolase, as well as the two receptors for leukotriene B(4) (BLT1R and BLT2R), corresponding to neutrophils in the luminal part of the thrombus. In contrast, in the vascular wall, the leukotriene pathway mainly localized in macrophage-rich adventitial areas. Furthermore, conditioned media of the intraluminal thrombus contained significantly higher concentrations of leukotriene B(4) than that derived from the vascular wall, which were significantly correlated to other neutrophil derived mediators, such as elastase/alpha(1)-antitrypsin complexes, myeloperoxidase, and MMP9/NGAL complexes. Finally, the neutrophil-chemotactic activity of the conditioned media from the intraluminal thrombus exhibited major inhibition by antagonists of the leukotriene B(4) receptors. Taken together, these results indicate neutrophil-derived leukotriene B(4) as a major neutrophil chemotactic factor released from the intraluminal thrombus of human AAAs and suggest that targeting BLT receptors may represent a potential medical therapeutic strategy in the prevention of AAA progression in humans. PMID- 19136616 TI - An acellular matrix-bound ligand enhances the mobilization, recruitment and therapeutic effects of circulating progenitor cells in a hindlimb ischemia model. AB - Circulating progenitor cells home to and engraft to sites of ischemia, mediated in part by the adhesion molecule L-selectin; however, accumulation in tissues such as the heart is low. In this study, an acellular collagen-based matrix containing sialyl Lewis(X) (sLe(X)), which binds L-selectin, was developed in order to enhance the endogenous progenitor cell therapeutic response. Its effect on progenitor cells and angiogenesis were assessed in vitro and using a hindlimb ischemia model with rats. In culture, the sLe(X)-collagen matrix recruited more CD133(+)CD34(+)L-selectin(+) cells than collagen-only matrix, with adhesion mediated by L-selectin binding. Increased angiogenic/chemotactic cytokine production and improved resistance to apoptosis appeared in cells cultured on sLe(X)-collagen matrix. In vivo, mobilization of endogenous circulating progenitor cells was increased, and greater recruitment of these and systemically injected human peripheral blood CXCR4(+)L-selectin(+) cells to sLe(X)-collagen treated limbs was observed compared to collagen-only. This condition was associated with differences in angiogenic/chemotactic cytokine levels, with greater arteriole density and increased perfusion in sLe(X)-collagen treated hindlimbs. With these factors taken together, we demonstrated that an acellular matrix-bound ligand approach can enhance the mobilization, recruitment, and therapeutic effects of endogenous and/or transplanted progenitor cells, possibly through paracrine and antiapoptotic mechanisms, and could be used to improve cell based regenerative therapies. PMID- 19136617 TI - Genome-wide identification of DNA-protein interactions using chromatin immunoprecipitation coupled with flow cell sequencing. AB - The transcriptional networks underlying mammalian cell development and function are largely unknown. The recently described use of flow cell sequencing devices in combination with chromatin immunoprecipitation (ChIP-seq) stands to revolutionize the identification of DNA-protein interactions. As such, ChIP-seq is rapidly becoming the method of choice for the genome-wide localization of histone modifications and transcription factor binding sites. As further studies are performed, the information generated by ChIP-seq is expected to allow the development of a framework for networks describing the transcriptional regulation of cellular development and function. However, to date, this technology has been applied only to a small number of cell types, and even fewer tissues, suggesting a huge potential for novel discovery in this field. PMID- 19136618 TI - Activins regulate 17beta-hydroxysteroid dehydrogenase type I transcription in murine gonadotrope cells. AB - Activins are pleiotropic members of the TGFbeta superfamily and were initially characterized based on their abilities to stimulate FSH synthesis and secretion by gonadotrope cells of the anterior pituitary gland. Here, we identified the gene encoding the steroidogenic enzyme, 17beta-hydroxysteroid dehydrogenase type I (17beta-HSD1; Hsd17b1), as an activin-responsive gene in immortalized gonadotrope cells, LbetaT2. 17beta-HSD1 catalyzes the conversion of estrone to the more active 17beta-estradiol, and activin A stimulated an increase in this enzymatic activity in these cells. We demonstrated that activins signaled via the type I receptor, activin receptor-like kinase (ALK4), and the intracellular signaling protein, SMAD2, to regulate Hsd17b1 transcription in immediate-early fashion. Critical cis-elements, including a minimal SMAD-binding element, were mapped to within 100 bp of the start of transcription. Activin/ALK4 signaling also regulated Hsd17b1 transcription in both immortalized and primary cultured murine granulosa cells. The promoter regions mediating basal and activin/ALK4 regulated promoter activity were generally conserved across the different cell types. The data show that activin A rapidly regulates Hsd17b1 transcription in gonadotrope and granulosa cells and may thereby regulate local 17beta-estradiol synthesis. PMID- 19136619 TI - A long-acting glucagon-like peptide-1 analogue attenuates induction of plasminogen activator inhibitor type-1 and vascular adhesion molecules. AB - Glucagon-like peptide-1 (GLP-1) administration attenuates endothelial cell dysfunction in diabetic patients and inhibits tumour necrosis factor alpha (TNF) mediated plasminogen activator inhibitor type-1 (PAI-1) induction in human vascular endothelial cells. The short half-life of GLP-1 mediated via degradation by the enzyme dipeptidyl peptidase 4 mandates the clinical use of long-acting GLP 1 analogues. The effects of a long-acting GLP-1 analogue on PAI-1 and vascular adhesion molecule expression in vascular endothelial cells are unknown. In this report, we demonstrate for the first time that the treatment with liraglutide, a long-acting GLP-1 analogue, inhibited TNF or hyperglycaemia-mediated induction of PAI-1, intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 mRNA and protein expression in a human vascular endothelial cell line. In addition, treatment attenuated TNF- or hyperglycaemia-mediated induction of the orphan nuclear receptor Nur77 mRNA expression. Taken together, these observations indicate that liraglutide inhibits TNF- or glucose-mediated induction of PAI-1 and vascular adhesion molecule expression, and this effect may involve the modulation of NUR77. These effects suggest that liraglutide may potentially improve the endothelial cell dysfunction associated with premature atherosclerosis identified in type 2 diabetic patients. PMID- 19136620 TI - Identification, localization, and regulation of passerine GnRH-I messenger RNA. AB - The neuropeptide GnRH-I is critical for the regulation of reproduction in all vertebrates. Study of the regulation of GnRH-I in passerine songbirds has been the focus of studies on subjects as diverse as photoperiodism, puberty, stress, nutrition, processing of auditory information, migration, global climate change, and evolutionary biology. Until now, analysis of GnRH-I in songbirds has been limited to measurement of immunoreactive peptide. Measurement of mRNA regulation has been impossible because of lack of knowledge of the GnRH gene sequence, despite many attempts in the last 20 years to identify it. Thus, the relative roles of environmental, social, physiological, and evolutionary influences upon passerine GnRH regulation have remained enigmatic. Here, we report the first cloning of GnRH-I cDNA from a songbird, Taeniopygia guttata, its localization and regulation. Although the homology of its translated precursor polypeptide between chicken GnRH-I precursor polypeptide was only 54%, zebra finch GnRH-I precursor contained an amino acid sequence that can be processed into chicken GnRH-I peptide (pEHWSYGLQPG-amide). In situ hybridization combined with immunocytochemistry showed co-localization of GnRH-I mRNA and immunoreactive peptide in the preoptic area of sexually mature birds. GnRH-I mRNA signal was greatly reduced in sexually immature birds. Ovary mass of female birds was positively correlated with GnRH-I mRNA level in the brain. These data will now permit molecular analysis of the regulation of songbird reproduction by physical, social, and physiological cues, along with fine scale analysis of selection pressures acting upon the reproductive system of songbirds. (244/250). PMID- 19136621 TI - Making synaptic plasticity and memory last: mechanisms of translational regulation. AB - Synaptic transmission in neurons is a measure of communication at synapses, the points of contact between axons and dendrites. The magnitude of synaptic transmission is a reflection of the strength of these synaptic connections, which in turn can be altered by the frequency with which the synapses are stimulated, the arrival of stimuli from other neurons in the appropriate temporal window, and by neurotrophic factors and neuromodulators. The ability of synapses to undergo lasting biochemical and morphological changes in response to these types of stimuli and neuromodulators is known as synaptic plasticity, which likely forms the cellular basis for learning and memory, although the relationship between any one form synaptic plasticity and a particular type of memory is unclear. RNA metabolism, particularly translational control at or near the synapse, is one process that controls long-lasting synaptic plasticity and, by extension, several types of memory formation and consolidation. Here, we review recent studies that reflect the importance and challenges of investigating the role of mRNA translation in synaptic plasticity and memory formation. PMID- 19136622 TI - Systemic regulation of starvation response in Caenorhabditis elegans. AB - When the supply of environmental nutrients is limited, multicellular animals can make both physiological and behavioral changes so as to cope with nutrient starvation. Although physiological and behavioral effects of starvation are well known, the mechanisms by which animals sense starvation systemically remain elusive. Furthermore, what constituent of food is sensed and how it modulates starvation response is still poorly understood. In this study, we use a starvation-hypersensitive mutant to identify molecules and mechanisms that modulate starvation signaling. We found that specific amino acids could suppress the starvation-induced death of gpb-2 mutants, and that MGL-1 and MGL-2, Caenorhabditis elegans homologs of metabotropic glutamate receptors, were involved. MGL-1 and MGL-2 acted in AIY and AIB neurons, respectively. Treatment with leucine suppressed starvation-induced stress resistance and life span extension in wild-type worms, and mutation of mgl-1 and mgl-2 abolished these effects of leucine. Taken together, our results suggest that metabotropic glutamate receptor homologs in AIY and AIB neuron may modulate a systemic starvation response, and that C. elegans senses specific amino acids as an anti hunger signal. PMID- 19136623 TI - Phosphorylation of Swi6/HP1 regulates transcriptional gene silencing at heterochromatin. AB - Heterochromatin protein 1 (HP1) recruits various effectors to heterochromatin for multiple functions, but its regulation is unclear. In fission yeast, a HP1 homolog Swi6 recruits SHREC, Epe1, and cohesin, which are involved in transcriptional gene silencing (TGS), transcriptional activation, and sister chromatid cohesion, respectively. We found that casein kinase II (CK2) phosphorylated Swi6. Loss of CK2-dependent Swi6 phosphorylation alleviated heterochromatic TGS without affecting heterochromatin structure. This was due to the inhibited recruitment of SHREC to heterochromatin, accompanied by an increase in Epe1. Interestingly, loss of phosphorylation did not affect cohesion. These results indicate that CK2-dependent Swi6 phosphorylation specifically controls TGS in heterochromatin. PMID- 19136624 TI - GLI1 is regulated through Smoothened-independent mechanisms in neoplastic pancreatic ducts and mediates PDAC cell survival and transformation. AB - Pancreatic ductal adenocarcinoma (PDAC) is characterized by the deregulation of the hedgehog signaling pathway. The Sonic Hedgehog ligand (Shh), absent in the normal pancreas, is highly expressed in pancreatic tumors and is sufficient to induce neoplastic precursor lesions in mouse models. We investigated the mechanism of Shh signaling in PDAC carcinogenesis by genetically ablating the canonical bottleneck of hedgehog signaling, the transmembrane protein Smoothened (Smo), in the pancreatic epithelium of PDAC-susceptible mice. We report that multistage development of PDAC tumors is not affected by the deletion of Smo in the pancreas, demonstrating that autocrine Shh-Ptch-Smo signaling is not required in pancreatic ductal cells for PDAC progression. However, the expression of Gli target genes is maintained in Smo-negative ducts, implicating alternative means of regulating Gli transcription in the neoplastic ductal epithelium. In PDAC tumor cells, we find that Gli transcription is decoupled from upstream Shh-Ptch Smo signaling and is regulated by TGF-beta and KRAS, and we show that Gli1 is required both for survival and for the KRAS-mediated transformed phenotype of cultured PDAC cancer cells. PMID- 19136625 TI - E2f3b plays an essential role in myogenic differentiation through isoform specific gene regulation. AB - Current models posit that E2F transcription factors can be divided into members that either activate or repress transcription, in part through collaboration with the retinoblastoma (pRb) tumor suppressor family. The E2f3 locus encodes E2f3a and E2f3b proteins, and available data suggest that they regulate cell cycle dependent gene expression through opposing transcriptional activating and repressing activities in growing and quiescent cells, respectively. However, the role, if any, of E2F proteins, and in particular E2f3, in myogenic differentiation is not well understood. Here, we dissect the contributions of E2f3 isoforms and other activating and repressing E2Fs to cell cycle exit and differentiation by performing genome-wide identification of isoform-specific targets. We show that E2f3a and E2f3b target genes are involved in cell growth, lipid metabolism, and differentiation in an isoform-specific manner. Remarkably, using gene silencing, we show that E2f3b, but not E2f3a or other E2F family members, is required for myogenic differentiation, and that this requirement for E2f3b does not depend on pRb. Our functional studies indicate that E2f3b specifically attenuates expression of genes required to promote differentiation. These data suggest how diverse E2F isoforms encoded by a single locus can play opposing roles in cell cycle exit and differentiation. PMID- 19136626 TI - Yeast Mph1 helicase dissociates Rad51-made D-loops: implications for crossover control in mitotic recombination. AB - Eukaryotes possess mechanisms to limit crossing over during homologous recombination, thus avoiding possible chromosomal rearrangements. We show here that budding yeast Mph1, an ortholog of human FancM helicase, utilizes its helicase activity to suppress spontaneous unequal sister chromatid exchanges and DNA double-strand break-induced chromosome crossovers. Since the efficiency and kinetics of break repair are unaffected, Mph1 appears to channel repair intermediates into a noncrossover pathway. Importantly, Mph1 works independently of two other helicases-Srs2 and Sgs1-that also attenuate crossing over. By chromatin immunoprecipitation, we find targeting of Mph1 to double-strand breaks in cells. Purified Mph1 binds D-loop structures and is particularly adept at unwinding these structures. Importantly, Mph1, but not a helicase-defective variant, dissociates Rad51-made D-loops. Overall, the results from our analyses suggest a new role of Mph1 in promoting the noncrossover repair of DNA double strand breaks. PMID- 19136627 TI - Second messenger-mediated spatiotemporal control of protein degradation regulates bacterial cell cycle progression. AB - Second messengers control a wide range of important cellular functions in eukaryotes and prokaryotes. Here we show that cyclic di-GMP, a global bacterial second messenger, promotes cell cycle progression in Caulobacter crescentus by mediating the specific degradation of the replication initiation inhibitor CtrA. During the G1-to-S-phase transition, both CtrA and its cognate protease ClpXP dynamically localize to the old cell pole, where CtrA is rapidly degraded. Sequestration of CtrA to the cell pole depends on PopA, a newly identified cyclic di-GMP effector protein. PopA itself localizes to the cell pole and directs CtrA to this subcellular site via the direct interaction with a mediator protein, RcdA. We present evidence that c-di-GMP regulates CtrA degradation during the cell cycle by controlling the dynamic sequestration of the PopA recruitment factor to the cell pole. Furthermore, we show that cell cycle timing of CtrA degradation relies on converging pathways responsible for substrate and protease localization to the old cell pole. This is the first report that links cyclic di GMP to protein dynamics and cell cycle control in bacteria. PMID- 19136628 TI - Transcription is required for establishment of germline methylation marks at imprinted genes. AB - Genomic imprinting requires the differential marking by DNA methylation of genes in male and female gametes. In the female germline, acquisition of methylation imprint marks depends upon the de novo methyltransferase Dnmt3a and its cofactor Dnmt3L, but the reasons why specific sequences are targets for Dnmt3a and Dnmt3L are still poorly understood. Here, we investigate the role of transcription in establishing maternal germline methylation marks. We show that at the Gnas locus, truncating transcripts from the furthest upstream Nesp promoter disrupts oocyte derived methylation of the differentially methylated regions (DMRs). Transcription through DMRs in oocytes is not restricted to this locus but occurs across the prospective DMRs at many other maternally marked imprinted domains, suggesting a common requirement for transcription events. The transcripts implicated here in gametic methylation are protein-coding, in contrast to the noncoding antisense transcripts involved in the monoallelic silencing of imprinted genes in somatic tissues, although they often initiate from alternative promoters in oocytes. We propose that transcription is a third essential component of the de novo methylation system, which includes optimal CpG spacing and histone modifications, and may be required to create or maintain open chromatin domains to allow the methylation complex access to its preferred targets. PMID- 19136629 TI - PRMT1-mediated arginine methylation of PIAS1 regulates STAT1 signaling. AB - To elucidate the function of the transcriptional coregulator PRMT1 (protein arginine methyltranferase 1) in interferon (IFN) signaling, we investigated the expression of STAT1 (signal transducer and activator of transcription) target genes in PRMT1-depleted cells. We show here that PRMT1 represses a subset of IFNgamma-inducible STAT1 target genes in a methyltransferase-dependent manner. These genes are also regulated by the STAT1 inhibitor PIAS1 (protein inhibitor of activated STAT1). PIAS1 is arginine methylated by PRMT1 in vitro as well as in vivo upon IFN treatment. Mutational and mass spectrometric analysis of PIAS1 identifies Arg 303 as the single methylation site. Using both methylation deficient and methylation-mimicking mutants, we find that arginine methylation of PIAS1 is essential for the repressive function of PRMT1 in IFN-dependent transcription and for the recruitment of PIAS1 to STAT1 target gene promoters in the late phase of the IFN response. Methylation-dependent promoter recruitment of PIAS1 results in the release of STAT1 and coincides with the decline of STAT1 activated transcription. Accordingly, knockdown of PRMT1 or PIAS1 enhances the anti-proliferative effect of IFNgamma. Our findings identify PRMT1 as a novel and crucial negative regulator of STAT1 activation that controls PIAS1-mediated repression by arginine methylation. PMID- 19136630 TI - Rapid determination of hydrogen positions and protonation states of diisopropyl fluorophosphatase by joint neutron and X-ray diffraction refinement. AB - Hydrogen atoms constitute about half of all atoms in proteins and play a critical role in enzyme mechanisms and macromolecular and solvent structure. Hydrogen atom positions can readily be determined by neutron diffraction, and as such, neutron diffraction is an invaluable tool for elucidating molecular mechanisms. Joint refinement of neutron and X-ray diffraction data can lead to improved models compared with the use of neutron data alone and has now been incorporated into modern, maximum-likelihood based crystallographic refinement programs like CNS. Joint refinement has been applied to neutron and X-ray diffraction data collected on crystals of diisopropyl fluorophosphatase (DFPase), a calcium-dependent phosphotriesterase capable of detoxifying organophosphorus nerve agents. Neutron omit maps reveal a number of important features pertaining to the mechanism of DFPase. Solvent molecule W33, coordinating the catalytic calcium, is a water molecule in a strained coordination environment, and not a hydroxide. The smallest Ca-O-H angle is 53 degrees, well beyond the smallest angles previously observed. Residue Asp-229, is deprotonated, supporting a mechanism involving nucleophilic attack by Asp-229, and excluding water activation by the catalytic calcium. The extended network of hydrogen bonding interactions in the central water filled tunnel of DFPase is revealed, showing that internal solvent molecules form an important, integrated part of the overall structure. PMID- 19136631 TI - CD40-CD40L cross-talk integrates strong antigenic signals and microbial stimuli to induce development of IL-17-producing CD4+ T cells. AB - IL-17-producing CD4(+) T cells have been recognized as key players in organ related autoimmune disease; however, the parameters that govern their development are yet to be elucidated fully. By using both in vivo and in vitro systems, we have investigated the role of antigen dose, pathogen-associated molecular patterns, and CD40-CD40 ligand (CD40L) cross-talk in Th17 differentiation. We found that the strength of antigenic stimulation critically influenced the extent of Th17 differentiation, because high, but not low or intermediate, antigen concentrations led to IL-17 production. Strong antigenic stimulation of T cells up-regulated CD40L expression, which in concert with certain microbial stimuli (i.e., cytosine phosphate guanine, curdlan, and zymosan) synergistically increased dendritic cell (DC) IL-6 production and Th17 polarization. CD40 deficient DCs exhibited reduced cytokine release and failed to drive Th17 development in vitro. These results were confirmed in vivo where the absence of CD40-CD40L cross-talk was found to prevent the expansion of IL-17-producing cells and accordingly the development of experimental autoimmune encephalitis. Our data demonstrate that CD40-CD40L cross-talk is important for Th17 development by translating strong T cell receptor and microbial stimuli into IL-6 production. PMID- 19136632 TI - The tumor suppressor Cdc73 functionally associates with CPSF and CstF 3' mRNA processing factors. AB - The CDC73 tumor suppressor gene is mutationally inactivated in hereditary and sporadic parathyroid tumors. Its product, the Cdc73 protein, is a component of the RNA polymerase II and chromatin-associated human Paf1 complex (Paf1C). Here, we show that Cdc73 physically associates with the cleavage and polyadenylation specificity factor (CPSF) and cleavage stimulation factor (CstF) complexes that are required for the maturation of mRNA 3' ends in the cell nucleus. Immunodepletion experiments indicate that the Cdc73-CPSF-CstF complex is necessary for 3' mRNA processing in vitro. Microarray analysis of CDC73 siRNA treated cells revealed INTS6, a gene encoding a subunit of the Integrator complex, as an in vivo Cdc73 target. Cdc73 depletion by siRNA resulted in decreased INTS6 mRNA abundance, and decreased association of CPSF and CstF subunits with the INTS6 locus. Our results suggest that Cdc73 facilitates association of 3' mRNA processing factors with actively-transcribed chromatin and support the importance of links between tumor suppression and mRNA maturation. PMID- 19136633 TI - PTMap--a sequence alignment software for unrestricted, accurate, and full spectrum identification of post-translational modification sites. AB - We present sequence alignment software, called PTMap, for the accurate identification of full-spectrum protein post-translational modifications (PTMs) and polymorphisms. The software incorporates several features to improve searching speed and accuracy, including peak selection, adjustment of inaccurate mass shifts, and precise localization of PTM sites. PTMap also automates rules, based mainly on unmatched peaks, for manual verification of identified peptides. To evaluate the quality of sequence alignment, we developed a scoring system that takes into account both matched and unmatched peaks in the mass spectrum. Incorporation of these features dramatically increased both accuracy and sensitivity of the peptide- and PTM-identifications. To our knowledge, PTMap is the first algorithm that emphasizes unmatched peaks to eliminate false positives. The superior performance and reliability of PTMap were demonstrated by confident identification of PTMs on 156 peptides from four proteins and validated by MS/MS of the synthetic peptides. Our results demonstrate that PTMap is a powerful algorithm capable of identification of all possible protein PTMs with high confidence. PMID- 19136634 TI - Insulin levels control female germline stem cell maintenance via the niche in Drosophila. AB - Stem cell maintenance depends on local signals provided by specialized microenvironments, or niches, in which they reside. The potential role of systemic factors in stem cell maintenance, however, has remained largely unexplored. Here, we show that insulin signaling integrates the effects of diet and age on germline stem cell (GSC) maintenance through the dual regulation of cap cell number (via Notch signaling) and cap cell-GSC interaction (via E cadherin) and that the normal process of GSC and niche cell loss that occurs with age can be suppressed by increased levels of insulin-like peptides. These results underscore the importance of systemic factors for the regulation of stem cell niches and, thereby, of stem cell numbers. PMID- 19136635 TI - Basis for the preferential activation of cardiac sympathetic nerve activity in heart failure. AB - In heart failure (HF), sympathetic nerve activity is increased. Measurements in HF patients of cardiac norepinephrine spillover, reflecting cardiac sympathetic nerve activity (CSNA), indicate that it is increased earlier and to a greater extent than sympathetic activity to other organs. This has important consequences because it worsens prognosis, provoking arrhythmias and sudden death. To elucidate the mechanisms responsible for the activation of CSNA in HF, we made simultaneous direct neural recordings of CSNA and renal SNA (RSNA) in two groups of conscious sheep: normal animals and animals in HF induced by chronic, rapid ventricular pacing. In normal animals, the level of activity, measured as burst incidence (bursts of pulse related activity/100 heart beats), was significantly lower for CSNA (30 +/- 5%) than for RSNA (94 +/- 2%). Furthermore, the resting level of CSNA, relative to its maximum achieved while baroreceptors were unloaded by reducing arterial pressure, was set at a much lower percentage than RSNA. In HF, burst incidence of CSNA increased from 30 to 91%, whereas burst incidence of RSNA remained unaltered at 95%. The sensitivity of the control of both CSNA and RSNA by the arterial baroreflex remained unchanged in HF. These data show that, in the normal state, the resting level of CSNA is set at a lower level than RSNA, but in HF, the resting levels of SNA to both organs are close to their maxima. This finding provides an explanation for the preferential increase in cardiac norepinephrine spillover observed in HF. PMID- 19136636 TI - 3D structure of the C3bB complex provides insights into the activation and regulation of the complement alternative pathway convertase. AB - Generation of the alternative pathway C3-convertase, the central amplification enzyme of the complement cascade, initiates by the binding of factor B (fB) to C3b to form the proconvertase, C3bB. C3bB is subsequently cleaved by factor D (fD) at a single site in fB, producing Ba and Bb fragments. Ba dissociates from the complex, while Bb remains bound to C3b, forming the active alternative pathway convertase, C3bBb. Using single-particle electron microscopy we have determined the 3-dimensional structures of the C3bB and the C3bBb complexes at approximately 27A resolution. The C3bB structure shows that fB undergoes a dramatic conformational change upon binding to C3b. However, the C3b-bound fB structure was easily interpreted after independently fitting the atomic structures of the isolated Bb and Ba fragments. Interestingly, the divalent cation-binding site in the von Willebrand type A domain in Bb faces the C345C domain of C3b, whereas the serine-protease domain of Bb points outwards. The structure also shows that the Ba fragment interacts with C3b separately from Bb at the level of the alpha'NT and CUB domains. Within this conformation, the long and flexible linker between Bb and Ba is likely exposed and accessible for cleavage by fD to form the active convertase, C3bBb. The architecture of the C3bB and C3bBb complexes reveals that C3b could promote cleavage and activation of fB by actively displacing the Ba domain from the von Willebrand type A domain in free fB. These structures provide a structural basis to understand fundamental aspects of the activation and regulation of the alternative pathway C3 convertase. PMID- 19136637 TI - Pharmacological characterization of ATPM [(-)-3-aminothiazolo[5,4-b]-N cyclopropylmethylmorphinan hydrochloride], a novel mixed kappa-agonist and mu agonist/-antagonist that attenuates morphine antinociceptive tolerance and heroin self-administration behavior. AB - ATPM [(-)-3-amino-thiazolo[5,4-b]-N-cyclopropylmethylmorphinan hydrochloride] was found to have mixed kappa- and mu-opioid activity and identified to act as a full kappa-agonist and a partial mu-agonist by in vitro binding assays. The present study was undertaken to characterize its in vivo effects on morphine antinociceptive tolerance in mice and heroin self-administration in rats. ATPM was demonstrated to yield more potent antinociceptive effects than (-)U50,488H (trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl]benzeneacetamide). It was further found that the antinociceptive effects of ATPM were mediated by kappa- and mu-, but not delta-opioid, receptors. In addition to its agonist profile on the mu-receptor, ATPM also acted as a mu-antagonist, as measured by its inhibition of morphine-induced antinociception. It is more important that ATPM had a greater ratio of the ED(50) value of sedation to that of antinociception than (-)U50,488 (11.8 versus 3.7), indicative of a less sedative effect than (-)U50,488H. In addition, ATPM showed less potential to develop antinociceptive tolerance relative to (-)U50,488H and morphine. Moreover, it dose dependently inhibited morphine-induced antinociceptive tolerance. Furthermore, it was found that chronic treatment of rats for 8 consecutive days with ATPM (0.5 mg/kg s.c.) produced sustained decreases in heroin self-administration. ( )U50,488H (2 mg/kg s.c.) also produced similar inhibitory effect. Taken together, our findings demonstrated that ATPM, a novel mixed kappa-agonist and mu-agonist/ antagonist, could inhibit morphine-induced antinociceptive tolerance, with less potential to develop tolerance and reduce heroin self-administration with less sedative effect. kappa-Agonists with some mu-activity appear to offer some advantages over selective kappa-agonists for the treatment of heroin abuse. PMID- 19136639 TI - Fluorescent ligands of the bradykinin B1 receptors: pharmacologic characterization and application to the study of agonist-induced receptor translocation and cell surface receptor expression. AB - Unlike the widely distributed and preformed B(2) receptors, the bradykinin B(1) receptors exhibit a highly regulated expression and minimal agonist-induced endocytosis. To evaluate the potential usefulness of fluorescent B(1) receptor probes applicable to live cell microscopy and cytofluorometry, combined chemical synthesis and pharmacologic evaluation have been conducted on novel 5(6) carboxyfluorescein [5(6)CF]-containing peptides. Representative agents are the antagonist B-10376 [5(6)CF-epsilon-aminocaproyl-Lys-Lys-[Hyp(3), CpG(5), D Tic(7), CpG(8)]des-Arg(9)-bradykinin] and the agonist B-10378 [5(6)CF-epsilon aminocaproyl-Lys-des-Arg(9)-bradykinin]. B-10376 has a K(i) of 10 to 20 nM to displace [(3)H]Lys-des-Arg(9)-bradykinin from rabbit or human recombinant B(1) receptors expressed in human embryonic kidney (HEK) 293 cells and is a surmountable antagonist in the rabbit aorta contractility assay (pA(2), 7.49). B 10378 was a full agonist at the naturally expressed B(1) receptor (rabbit aorta contraction, calcium transients in human smooth muscle cells) and had a binding competition K(i) of 19 or 89 nM at the recombinant rabbit or human receptor, respectively. Both fluorescent probes can label with specificity human or rabbit B(1) receptors expressed in HEK 293 cells (epifluorescence or confocal microscopy), but the agonist was associated with discontinuous plasma membrane labeling, which coincided with that of a red-emitting caveolin-1 conjugate. Cytofluorometry with B-10376 was applied to recombinant and, in human vascular smooth muscle cells, to naturally expressed B(1) receptors. In all fluorescent applications, the specific labeling was reduced by an excess of a B(1) receptor nonpeptide antagonist. Despite the loss of affinity determined by the introduction of a fluorophore in B(1) receptor agonist or antagonist peptides, the resulting agents allow original applications (imaging in live cells, cytofluorometry). PMID- 19136638 TI - Proerectile effects of dopamine D2-like agonists are mediated by the D3 receptor in rats and mice. AB - Dopamine D(2)-like agonists induce penile erection (PE) and yawning in a variety of species, effects that have been suggested recently to be specifically mediated by the D(4) and D(3) receptors, respectively. The current studies were aimed at characterizing a series of D(2), D(3), and D(4) agonists with respect to their capacity to induce PE and yawning in the rat and the proerectile effects of apomorphine [(R)-(-)-5,6,6a,7-tetrahydro-6-methyl-4H-dibenzo-[de,g]quinoline 10,11-diol hydrochloride] in wild-type and D(4) receptor (R) knockout (KO) mice. All D(3) agonists induced dose-dependent increases in PE and yawning over a similar range of doses, whereas significant increases in PE or yawning were not observed with any of the D(4) agonists. Likewise, D(2), D(3), and D(4) antagonists were assessed for their capacity to alter apomorphine- and pramipexole (N'-propyl-4,5,6,7-tetrahydrobenzothiazole-2,6-diamine dihydrochloride)-induced PE and yawning. The D(3) antagonist, PG01037 [N-{4-[4 (2,3-dichlorophenyl)-piperazin-1-yl]-trans-but-2-enyl}-4-pyridine-2-yl-benzamide hydrochloride], inhibited the induction of PE and yawning, whereas the D(2) antagonist, L-741,626 [3-[4-(4-chlorophenyl)-4-hydroxypiperidin-l-yl]methyl-1H indole], reversed the inhibition of PE and yawning observed at higher doses. The D(4) antagonist, L-745,870 [3-(4-[4-chlorophenyl]piperazin-1-yl)-methyl-1H pyrrolo[2,3-b]pyridine trihydrochloride], did not alter apomorphine- or pramipexole-induced PE or yawning. A role for the D(3) receptor was further supported because apomorphine was equipotent at inducing PE in wild-type and D(4)RKO mice, effects that were inhibited by the D(3) antagonist, PG01037, in both wild-type and D(4)R KO mice. Together, these studies provide strong support that D(2)-like agonist-induced PE and yawning are differentially mediated by the D(3) (induction) and D(2) (inhibition) receptors. These studies fail to support a role for the D(4) receptor in the regulation of PE or yawning by D(2)-like agonists. PMID- 19136640 TI - Rapid identification of the hepatic cytochrome P450 2C19 activity using a novel and noninvasive [13C]pantoprazole breath test. AB - We tested the hypothesis that the stable isotope [(13)C]pantoprazole is O demethylated by cytochrome P450 CYP2C19 and that the (13)CO(2) produced and exhaled in breath as a result can serve as a safe, rapid, and noninvasive phenotyping marker of CYP2C19 activity in vivo. Healthy volunteers who had been genotyped for the CYP2C19(*)2, CYP2C19(*)3, and CYP2C19(*)17 alleles were administered a single oral dose of [(13)C]pantoprazole sodium-sesquihydrate (100 mg) with 2.1 g of sodium bicarbonate. Exhaled (13)CO(2) and (12)CO(2) were measured by IR spectroscopy before (baseline) and 2.5 to 120 min after dosing. Ratios of (13)CO(2)/(12)CO(2) after [(13)C]pantoprazole relative to (13)CO(2)/(12)CO(2) at baseline were expressed as change over baseline (DOB). Maximal DOB, DOB(15) to DOB(120), and area under the DOB versus time curve (AUC(0 120) and AUC(0-infinity)) were significantly different among three genotype groups (CYP2C19(*)1/(*)1, n = 10; CYP2C19(*)1/(*)2 or CYP2C19(*)1/(*)3, n = 10; and CYP2C19(*)2/(*)2, n = 5) with predicted extensive metabolizers (EMs), intermediate metabolizers (IMs), and poor metabolizers (PMs) of CYP2C19, respectively (Kruskal-Wallis test, p < 0.01); linear regression analysis indicated a gene-dose effect relationship (r(2) ranged between 0.236 and 0.522; all p < 0.05). These breath test indices were significantly lower in PMs than IMs (p < 0.05) or EMs (p < 0.01) of CYP2C19. [(13)C]Pantoprazole plasma exposure showed significant inverse correlation with breath test indices in the respective subjects (Pearson r = -0.74; p = 0.038). These feasibility data suggest that the [(13)C]pantoprazole breath test is a reliable, rapid, and noninvasive probe of CYP2C19 and seems to be a useful tool to optimize drug therapy metabolized by CYP2C19. PMID- 19136641 TI - An educational program in a pediatric hospice setting. AB - Canuck Place Children's Hospice, a family-centered pediatric hospice in Vancouver, Canada, provides family support and respite, pain and symptom management, and end-of-life care. One of the goals of pediatric hospice palliative care is to create an environment that supports a normal way of life and enhances quality of life. At Canuck Place, a unique school program for children with progressive life-threatening illnesses has been set up to meet this goal. This article describes the Canuck Place educational program, gives insights into the importance and challenges of providing a complete school experience, and discusses the expanded role of the teacher in the pediatric hospice setting. PMID- 19136642 TI - Beyond polarization, public preferences suggest policy opportunities to address aging, dying, and family caregiving. AB - Despite well-documented deficiencies and widespread suffering experienced by millions of elderly or ill Americans and their families, politicians rarely address end-of-life issues. Citizen Forums in New Hampshire surveyed 463 people regarding aging, serious illness, and caregiving. More than 80% indicated it was very or extremely important to have their dignity respected, preferences honored, pain controlled, and to not leave family with debt. Less than half strongly endorsed being kept alive as long as possible, prayed with or for, or having assisted-suicide available. Over 80% strongly endorsed palliative care requirements clinical licensure and reimbursement, expansion of family caregiver leave, respite care, and bereavement support. By avoiding actions which elicit strong divergence of opinion and focusing on actions on which consensus exists, public officials and candidates can respond to problems and improve care and experience for frail elders, dying Americans, and their families. PMID- 19136643 TI - Severe hemorrhage from a hypodermoclysis site. AB - Hypodermoclysis, or subcutaneous infusion of fluids, is an alternative means of administration of fluid. It is sometimes used in older patients in whom there is difficulty achieving intravenous access, or who are entering the terminal stage. It is considered a relatively low-risk procedure. We report a case where a patient's death may have resulted from its use. PMID- 19136644 TI - A plastidial pathway for protein isoprenylation in tobacco cells. PMID- 19136645 TI - Rearrangement of actin cytoskeleton mediates invasion of Lotus japonicus roots by Mesorhizobium loti. AB - Infection thread-dependent invasion of legume roots by rhizobia leads to internalization of bacteria into the plant cells, which is one of the salient features of root nodule symbiosis. We found that two genes, Nap1 (for Nck associated protein 1) and Pir1 (for 121F-specific p53 inducible RNA), involved in actin rearrangements were essential for infection thread formation and colonization of Lotus japonicus roots by its natural microsymbiont, Mesorhizobium loti. nap1 and pir1 mutants developed an excess of uncolonized nodule primordia, indicating that these two genes were not essential for the initiation of nodule organogenesis per se. However, both the formation and subsequent progression of infection threads into the root cortex were significantly impaired in these mutants. We demonstrate that these infection defects were due to disturbed actin cytoskeleton organization. Short root hairs of the mutants had mostly transverse or web-like actin filaments, while bundles of actin filaments in wild-type root hairs were predominantly longitudinal. Corroborating these observations, temporal and spatial differences in actin filament organization between wild-type and mutant root hairs were also observed after Nod factor treatment, while calcium influx and spiking appeared unperturbed. Together with various effects on plant growth and seed formation, the nap1 and pir1 alleles also conferred a characteristic distorted trichome phenotype, suggesting a more general role for Nap1 and Pir1 in processes establishing cell polarity or polar growth in L. japonicus. PMID- 19136646 TI - The Arabidopsis MYB5 transcription factor regulates mucilage synthesis, seed coat development, and trichome morphogenesis. AB - The Arabidopsis thaliana MYB5 gene is expressed in trichomes and seeds, including the seed coat. Constitutive expression of MYB5 resulted in the formation of more small trichomes and ectopic trichomes and a reduction in total leaf trichome numbers and branching. A myb5 mutant displayed minimal changes in trichome morphology, while a myb23 mutant produced increased numbers of small trichomes and two-branched trichomes. A myb5 myb23 double mutant developed more small rosette trichomes and two-branched trichomes than the single mutants. These results indicate that MYB5 and MYB23 regulate trichome extension and branching. The seed coat epidermal cells of myb5 and myb5 myb23 were irregular in shape, developed flattened columellae, and produced less mucilage than those of the wild type. Among the downregulated genes identified in the myb5 seeds using microarray analysis were ABE1 and ABE4 (alpha/beta fold hydrolase/esterase genes), MYBL2, and GLABRA2. The same genes were also downregulated in transparent testa glabra1 (ttg1) seeds, suggesting that MYB5 collaborates with TTG1 in seed coat development. These genes were upregulated in leaves and roots by ectopically expressed MYB5. The MYBL2, ABE1, and ABE4 promoters were active in seeds, including seed coats, and the latter two also in trichomes. Models of the MYB5 regulatory networks involved in seed coat and trichome development are presented. PMID- 19136647 TI - The plastidial 2-C-methyl-D-erythritol 4-phosphate pathway provides the isoprenyl moiety for protein geranylgeranylation in tobacco BY-2 cells. AB - Protein farnesylation and geranylgeranylation are important posttranslational modifications in eukaryotic cells. We visualized in transformed Nicotiana tabacum Bright Yellow-2 (BY-2) cells the geranylgeranylation and plasma membrane localization of GFP-BD-CVIL, which consists of green fluorescent protein (GFP) fused to the C-terminal polybasic domain (BD) and CVIL isoprenylation motif from the Oryza sativa calmodulin, CaM61. Treatment with fosmidomycin (Fos) or oxoclomazone (OC), inhibitors of the plastidial 2-C-methyl-d-erythritol 4 phosphate (MEP) pathway, caused mislocalization of the protein to the nucleus, whereas treatment with mevinolin, an inhibitor of the cytosolic mevalonate pathway, did not. The nuclear localization of GFP-BD-CVIL in the presence of MEP pathway inhibitors was completely reversed by all-trans-geranylgeraniol (GGol). Furthermore, 1-deoxy-d-xylulose (DX) reversed the effects of OC, but not Fos, consistent with the hypothesis that OC blocks 1-deoxy-d-xylulose 5-phosphate synthesis, whereas Fos inhibits its conversion to 2-C-methyl-d-erythritol 4 phosphate. By contrast, GGol and DX did not rescue the nuclear mislocalization of GFP-BD-CVIL in the presence of a protein geranylgeranyltransferase type 1 inhibitor. Thus, the MEP pathway has an essential role in geranylgeranyl diphosphate (GGPP) biosynthesis and protein geranylgeranylation in BY-2 cells. GFP-BD-CVIL is a versatile tool for identifying pharmaceuticals and herbicides that interfere either with GGPP biosynthesis or with protein geranylgeranylation. PMID- 19136649 TI - Adipose overexpression of desnutrin promotes fatty acid use and attenuates diet induced obesity. AB - OBJECTIVE: To investigate the role of desnutrin in adipose tissue triacylglycerol (TAG) and fatty acid metabolism. RESEARCH DESIGN AND METHODS: We generated transgenic mice overexpressing desnutrin (also called adipose triglyceride lipase [ATGL]) in adipocytes (aP2-desnutrin) and also performed adenoviral-mediated overexpression of desnutrin in 3T3-L1CARDelta1 adipocytes. RESULTS: aP2-desnutrin mice were leaner with decreased adipose tissue TAG content and smaller adipocyte size. Overexpression of desnutrin increased lipolysis but did not result in increased serum nonesterified fatty acid levels or ectopic TAG storage. We found increased cycling between diacylglycerol (DAG) and TAG and increased fatty acid oxidation in adipocytes from these mice, as well as improved insulin sensitivity. CONCLUSIONS: We show that by increasing lipolysis, desnutrin overexpression causes reduced adipocyte TAG content and attenuation of diet-induced obesity. Desnutrin-mediated lipolysis promotes fatty acid oxidation and re-esterification within adipocytes. PMID- 19136648 TI - The functional role of pack-MULEs in rice inferred from purifying selection and expression profile. AB - Gene duplication is an important mechanism for evolution of new genes. In plants, a special group of transposable elements, called Pack-MULEs or transduplicates, is able to duplicate and amplify genes or gene fragments on a large scale. Despite the abundance of Pack-MULEs, the functionality of these duplicates is not clear. Here, we present a comprehensive analysis of expression and purifying selection on 2809 Pack-MULEs in rice (Oryza sativa), which are derived from 1501 parental genes. At least 22% of the Pack-MULEs are transcribed, and 28 Pack-MULEs have direct evidence of translation. Chimeric Pack-MULEs, which contain gene fragments from multiple genes, are much more frequently expressed than those derived only from a single gene. In addition, Pack-MULEs are frequently associated with small RNAs. The presence of these small RNAs is associated with a reduction in expression of both the Pack-MULEs and their parental genes. Furthermore, an assessment of the selection pressure on the Pack-MULEs using the ratio of nonsynonymous (Ka) and synonymous (Ks) substitution rates indicates that a considerable number of Pack-MULEs likely have been under selective constraint. The Ka/Ks values of Pack-MULE and parental gene pairs are lower among Pack-MULEs that are expressed in sense orientations. Taken together, our analysis suggests that a significant number of Pack-MULEs are expressed and subjected to purifying selection, and some are associated with small RNAs. Therefore, at least a subset of Pack-MULEs are likely functional and have great potential in regulating gene expression as well as providing novel coding capacities. PMID- 19136650 TI - Intranasal insulin ameliorates experimental diabetic neuropathy. AB - OBJECTIVE: We hypothesized that intranasal insulin (I-I) delivery targets the nervous system while avoiding potential adverse systemic effects when compared with subcutaneous insulin (S-I) for experimental streptozotocin-induced diabetic peripheral neuropathy (DPN). RESEARCH DESIGN AND METHODS: I-I or S-I at 0.87 IU daily or placebo were delivered in separate cohorts of diabetic and nondiabetic CD1 mice during 8 months of diabetes. Radiolabeled insulin detection was used to compare delivery and biodistribution for I-I and S-I. Biweekly behavioral testing and monthly electrophysiological and quantitative studies assessed progression of DPN. At and before end point, morphometric analysis of DRG, peripheral nerve, distal epidermal innervation, and specific molecular markers were evaluated. RESULTS: Radiolabeled I-I resulted in more rapid and concentrated delivery to the spinal cord and DRG with less systemic insulin exposure. When compared with S-I or intranasal placebo, I-I reduced overall mouse mortality and sensory loss while improving neuropathic pain and electrophysiological/morphological abnormalities in diabetic mice. I-I restored mRNA and protein levels of phosphoinositide 3 kinase/Akt, cyclic AMP response element-binding protein, and glycogen synthase kinase 3beta to near normal levels within diabetic DRGs. CONCLUSIONS: I-I slows the progression of experimental DPN in streptozotocin mice, avoids adverse effects associated with S-I treatment, and prolongs lifespan when compared with S I. I-I may be a promising approach for the treatment of DPN. PMID- 19136651 TI - Pharmacological stimulation of NADH oxidation ameliorates obesity and related phenotypes in mice. AB - OBJECTIVE: Nicotinamide adenine dinucleotides (NAD+ and NADH) play a crucial role in cellular energy metabolism, and a dysregulated NAD+-to-NADH ratio is implicated in metabolic syndrome. However, it is still unknown whether a modulating intracellular NAD+-to-NADH ratio is beneficial in treating metabolic syndrome. We tried to determine whether pharmacological stimulation of NADH oxidation provides therapeutic effects in rodent models of metabolic syndrome. RESEARCH DESIGN AND METHODS: We used beta-lapachone (betaL), a natural substrate of NADH:quinone oxidoreductase 1 (NQO1), to stimulate NADH oxidation. The betaL induced pharmacological effect on cellular energy metabolism was evaluated in cells derived from NQO1-deficient mice. In vivo therapeutic effects of betaL on metabolic syndrome were examined in diet-induced obesity (DIO) and ob/ob mice. RESULTS: NQO1-dependent NADH oxidation by betaL strongly provoked mitochondrial fatty acid oxidation in vitro and in vivo. These effects were accompanied by activation of AMP-activated protein kinase and carnitine palmitoyltransferase and suppression of acetyl-coenzyme A (CoA) carboxylase activity. Consistently, systemic betaL administration in rodent models of metabolic syndrome dramatically ameliorated their key symptoms such as increased adiposity, glucose intolerance, dyslipidemia, and fatty liver. The treated mice also showed higher expressions of the genes related to mitochondrial energy metabolism (PPARgamma coactivator 1alpha, nuclear respiratory factor-1) and caloric restriction (Sirt1) consistent with the increased mitochondrial biogenesis and energy expenditure. CONCLUSIONS: Pharmacological activation of NADH oxidation by NQO1 resolves obesity and related phenotypes in mice, opening the possibility that it may provide the basis for a new therapy for the treatment of metabolic syndrome. PMID- 19136652 TI - Sex and depot differences in adipocyte insulin sensitivity and glucose metabolism. AB - OBJECTIVE: To investigate how insulin sensitivity and glucose metabolism differ in adipocytes between different fat depots of male and female mice and how sex steroids contribute to these differences. RESEARCH DESIGN AND METHODS: Adipocytes from intra-abdominal/perigonadal (PG) and subcutaneous (SC) adipose tissue from normal, castrated, or steroid-implanted animals were isolated and analyzed for differences in insulin sensitivity and glucose metabolism. RESULTS: Adipocytes from both PG and SC depots of females have increased lipogenic rates compared with those from males. In females, intra-abdominal PG adipocytes are more insulin sensitive than SC adipocytes and more insulin-sensitive than male adipocytes from either depot. When stimulated by low physiological concentrations of insulin, female PG adipocytes show a robust increase in Akt and extracellular signal related kinase (ERK) phosphorylation and lipogenesis, whereas male adipocytes show activation only at higher insulin concentrations. Adipocytes from females have higher mRNA/protein levels of several genes involved in glucose and lipid metabolism. After castration, adipocytes of male mice showed increased insulin sensitivity and increased lipogenic rates, whereas adipocytes of females demonstrate decreased lipid production. Increasing estrogen above physiological levels, however, also reduced lipid synthesis in females, whereas increasing dihydrotestosterone in males had no effect. CONCLUSIONS: There are major sex differences in insulin sensitivity in adipose tissue, particularly in the intra abdominal depot, that are regulated by physiological levels of sex steroids. The increased sensitivity to insulin and lipogenesis observed in adipocytes from females may account for their lower level of insulin resistance and diabetes risk despite similar or higher fat content than in males. PMID- 19136654 TI - Ciliary neurotrophic factor stimulates muscle glucose uptake by a PI3-kinase dependent pathway that is impaired with obesity. AB - OBJECTIVE: Ciliary neurotrophic factor (CNTF) reverses muscle insulin resistance by increasing fatty acid oxidation through gp130-LIF receptor signaling to the AMP-activated protein kinase (AMPK). CNTF also increases Akt signaling in neurons and adipocytes. Because both Akt and AMPK regulate glucose uptake, we investigated muscle glucose uptake in response to CNTF signaling in lean and obese mice. RESEARCH DESIGN AND METHODS: Mice were injected intraperitoneally with saline or CNTF, and blood glucose was monitored. The effects of CNTF on skeletal muscle glucose uptake and AMPK/Akt signaling were investigated in incubated soleus and extensor digitorum longus (EDL) muscles from muscle-specific AMPKalpha2 kinase-dead, gp130(DeltaSTAT), and lean and obese ob/ob and high-fat fed mice. The effect of C2-ceramide on glucose uptake and gp130 signaling was also examined. RESULTS: CNTF reduced blood glucose and increased glucose uptake in isolated muscles in a time- and dose-dependent manner with maximal effects after 30 min with 100 ng/ml. CNTF increased Akt-S473 phosphorylation in soleus and EDL; however, AMPK-T172 phosphorylation was only increased in soleus. Incubation of muscles from AMPK kinase dead (KD) and wild-type littermates with the PI3-kinase inhibitor LY-294002 demonstrated that PI3-kinase, but not AMPK, was essential for CNTF-stimulated glucose uptake. CNTF-stimulated glucose uptake and Akt phosphorylation were substantially reduced in obesity (high-fat diet and ob/ob) despite normal induction of gp130/AMPK signaling--effects also observed when treating myotubes with C2-ceramide. CONCLUSIONS: CNTF acutely increases muscle glucose uptake by a mechanism involving the PI3-kinase/Akt pathway that does not require AMPK. CNTF-stimulated glucose uptake is impaired in obesity induced insulin resistance and by ceramide. PMID- 19136653 TI - Survey of the human pancreatic beta-cell G1/S proteome reveals a potential therapeutic role for cdk-6 and cyclin D1 in enhancing human beta-cell replication and function in vivo. AB - OBJECTIVES: To comprehensively inventory the proteins that control the G1/S cell cycle checkpoint in the human islet and compare them with those in the murine islet, to determine whether these might therapeutically enhance human beta-cell replication, to determine whether human beta-cell replication can be demonstrated in an in vivo model, and to enhance human beta-cell function in vivo. RESEARCH DESIGN AND METHODS: Thirty-four G1/S regulatory proteins were examined in human islets. Effects of adenoviruses expressing cdk-6, cdk-4, and cyclin D1 on proliferation in human beta-cells were studied in both in vitro and in vivo models. RESULTS: Multiple differences between murine and human islets occur, most strikingly the presence of cdk-6 in human beta-cells versus its low abundance in the murine islet. Cdk-6 and cyclin D1 in vitro led to marked activation of retinoblastoma protein phosphorylation and cell cycle progression with no induction of cell death. Human islets transduced with cdk-6 and cyclin D1 were transplanted into diabetic NOD-SCID mice and markedly outperformed native human islets in vivo, maintaining glucose control for the entire 6 weeks of the study. CONCLUSIONS: The human G1/S proteome is described for the first time. Human islets are unlike their rodent counterparts in that they contain easily measurable cdk-6. Cdk-6 overexpression, alone or in combination with cyclin D1, strikingly stimulates human beta-cell replication, both in vitro as well as in vivo, without inducing cell death or loss of function. Using this model, human beta-cell replication can be induced and studied in vivo. PMID- 19136655 TI - Genome-wide scan for linkage to type 1 diabetes in 2,496 multiplex families from the Type 1 Diabetes Genetics Consortium. AB - OBJECTIVE: Type 1 diabetes arises from the actions of multiple genetic and environmental risk factors. Considerable success at identifying common genetic variants that contribute to type 1 diabetes risk has come from genetic association (primarily case-control) studies. However, such studies have limited power to detect genes containing multiple rare variants that contribute significantly to disease risk. RESEARCH DESIGN AND METHODS: The Type 1 Diabetes Genetics Consortium (T1DGC) has assembled a collection of 2,496 multiplex type 1 diabetic families from nine geographical regions containing 2,658 affected sib pairs (ASPs). We describe the results of a genome-wide scan for linkage to type 1 diabetes in the T1DGC family collection. RESULTS: Significant evidence of linkage to type 1 diabetes was confirmed at the HLA region on chromosome 6p21.3 (logarithm of odds [LOD] = 213.2). There was further evidence of linkage to type 1 diabetes on 6q that could not be accounted for by the major linkage signal at the HLA class II loci on chromosome 6p21. Suggestive evidence of linkage (LOD > or =2.2) was observed near CTLA4 on chromosome 2q32.3 (LOD = 3.28) and near INS (LOD = 3.16) on chromosome 11p15.5. Some evidence for linkage was also detected at two regions on chromosome 19 (LOD = 2.84 and 2.54). CONCLUSIONS: Five non-HLA chromosome regions showed some evidence of linkage to type 1 diabetes. A number of previously proposed type 1 diabetes susceptibility loci, based on smaller ASP numbers, showed limited or no evidence of linkage to disease. Low-frequency susceptibility variants or clusters of loci with common alleles could contribute to the linkage signals observed. PMID- 19136656 TI - Beta-Cell hyperplasia induced by hepatic insulin resistance: role of a liver pancreas endocrine axis through insulin receptor A isoform. AB - OBJECTIVE: Type 2 diabetes results from a combination of insulin resistance and impaired insulin secretion. To directly address the effects of hepatic insulin resistance in adult animals, we developed an inducible liver-specific insulin receptor knockout mouse (iLIRKO). RESEARCH DESIGN AND METHODS: Using this approach, we were able to induce variable insulin receptor (IR) deficiency in a tissue-specific manner (liver mosaicism). RESULTS: iLIRKO mice presented progressive hepatic and extrahepatic insulin resistance without liver dysfunction. Initially, iLIRKO mice displayed hyperinsulinemia and increased beta cell mass, the extent of which was proportional to the deletion of hepatic IR. Our studies of iLIRKO suggest a cause-and-effect relationship between progressive insulin resistance and the fold increase of plasma insulin levels and beta-cell mass. Ultimately, the beta-cells failed to secrete sufficient insulin, leading to uncontrolled diabetes. We observed that hepatic IGF-1 expression was enhanced in iLIRKO mice, resulting in an increase of circulating IGF-1. Concurrently, the IR A isoform was upregulated in hyperplastic beta-cells of iLIRKO mice and IGF-1 induced proliferation was higher than in the controls. In mouse beta-cell lines, IR-A, but not IR-B, conferred a proliferative capacity in response to insulin or IGF-1, providing a potential explanation for the beta-cell hyperplasia induced by liver insulin resistance in iLIRKO mice. CONCLUSIONS: Our studies of iLIRKO mice suggest a liver-pancreas endocrine axis in which IGF-1 functions as a liver derived growth factor to promote compensatory pancreatic islet hyperplasia through IR-A. PMID- 19136657 TI - Location of arterial stiffening differs in those with impaired fasting glucose versus diabetes: implications for left ventricular hypertrophy from the Multi Ethnic Study of Atherosclerosis. AB - OBJECTIVE: To determine whether middle-aged and older individuals with impaired fasting glucose (IFG), but no clinical evidence of cardiovascular disease, exhibit abnormal changes in proximal thoracic aortic stiffness or left ventricular (LV) mass when compared with healthy counterparts. RESEARCH DESIGN AND METHODS: From the Multi-Ethnic Study of Atherosclerosis, 2,240 subjects with normal fasting glucose (NFG), 845 with IFG, and 414 with diabetes, all aged 45 to 85 years and without preexisting coronary artery disease, underwent MRI determinations of total arterial and proximal thoracic aortic stiffness and LV mass. The presence or absence of other factors known to influence arterial stiffness was assessed. RESULTS: After adjustment for clinical factors known to modify arterial stiffness, proximal thoracic aortic stiffness was not increased in those with IFG compared with those with NFG (1.90 +/- 0.05 versus 1.91 +/- 0.04 10(-3) mmHg(-1), respectively, P = 0.83). After accounting for clinical factors known to influence LV mass, LV mass was increased in those with diabetes relative to those with NFG (150.6 +/- 1.4 versus 145.8 +/- 0.81 g, P < 0.0009) but not in those with IFG in comparison with NFG (145.2 +/- 1.03 versus 145.8 +/- 0.81 g, P = 0.56). CONCLUSIONS: Middle-aged and older individuals with the pre diabetes state of IFG do not exhibit abnormal proximal thoracic distensibility or LV hypertrophy relative to individuals with NFG. For this reason, an opportunity may exist in those with IFG to prevent LV hypertrophy and abnormal aortic stiffness that is observed in middle-aged and older individuals with diabetes. PMID- 19136658 TI - Relationship of metabolic syndrome with incident aortic valve calcium and aortic valve calcium progression: the Multi-Ethnic Study of Atherosclerosis (MESA). AB - OBJECTIVE: Metabolic syndrome (MetS) has been associated with increased prevalence of aortic valve calcium (AVC) and with increased progression of aortic stenosis. The purpose of this study was to determine whether MetS is associated with increased risks for the development of new ("incident") AVC or for progression of established AVC as assessed by CT. RESEARCH DESIGN AND METHODS: The relationships of MetS or its components as well as of diabetes to risks for incident AVC or AVC progression were studied among participants with CT scans performed at baseline and at either year 2 or year 3 examinations in the Multi Ethnic Study of Atherosclerosis (MESA). RESULTS: Of 5,723 MESA participants meeting criteria for inclusion, 1,674 had MetS by Adult Treatment Panel III criteria, whereas 761 had diabetes. Among the 5,123 participants without baseline AVC, risks for incident AVC, adjusted for time between scans, age, sex, race/ethnicity, LDL cholesterol, lipid-lowering medications, and smoking, were increased significantly for MetS (odds ratio [OR] 1.67 [95% CI 1.21-2.31]) or diabetes (2.06 [1.39-3.06]). In addition, there was an increase in incident AVC risk with increasing number of MetS components. Similar results were found using the International Diabetes Federation MetS criteria. Among the 600 participants (10.5%) with baseline AVC, neither MetS nor diabetes was associated with AVC progression. CONCLUSIONS: In the MESA cohort, MetS was associated with a significant increase in incident ("new") AVC, raising the possibility that MetS may be a potential therapeutic target to prevent AVC development. PMID- 19136659 TI - SHIP limits immunoregulatory capacity in the T-cell compartment. AB - Regulatory T cells (T(regs)) play a pivotal role in preventing autoimmunity, graft-versus-host disease (GVHD), and organ graft rejection. We previously showed that either germline or induced SH2 domain-containing inositol 5-phosphatase (SHIP) deficiency in the host abrogates GVHD. Here we show that SHIP deficiency promotes an increase of CD4(+)CD25(+)FoxP3(+) T(regs) and CD4(+)CD25( )FoxP3(+)"naive" T cells in the periphery that display increased CD103, glucocorticoid-induced tumor necrosis factor receptor-related protein (GITR), OX40, and FcgammaRII/III expression. SHIP deficiency does not compromise T(reg) function because SHIP-deficient CD3(+)CD4(+)CD25(+) T(regs) are as suppressive as wild-type (WT) CD3(+)CD4(+)CD25(+) T(reg). Interestingly, like conventional T(regs), SHIP(-/-) CD4(+)CD25(-) T cells are unresponsive to major histocompatibility complex (MHC)-mismatched stimulators and suppress allogeneic responses by T cells in vitro. In addition, SHIP(-/-) CD4(+)CD25(-) T cells mediate reduced lethal GVHD on adoptive transfer to MHC-mismatched hosts. Furthermore, hosts with induced SHIP deficiency exhibit delayed rejection of MHC mismatched cardiac grafts. Thus, SHIP is required for robust graft-versus-host and host-versus-graft responses by CD4(+) T cell and limits their immunoregulatory capacity. These findings further define the immunosuppressive mechanisms that result from SHIP deficiency and provide additional justification for targeting SHIP in clinical transplantation. PMID- 19136660 TI - Role for MKL1 in megakaryocytic maturation. AB - Megakaryoblastic leukemia 1 (MKL1), identified as part of the t(1;22) translocation specific to acute megakaryoblastic leukemia, is highly expressed in differentiated muscle cells and promotes muscle differentiation by activating serum response factor (SRF). Here we show that Mkl1 expression is up-regulated during murine megakaryocytic differentiation and that enforced overexpression of MKL1 enhances megakaryocytic differentiation. When the human erythroleukemia (HEL) cell line is induced to differentiate with 12-O-tetradecanoylphorbol 13 acetate, overexpression of MKL1 results in an increased number of megakaryocytes with a concurrent increase in ploidy. MKL1 overexpression also promotes megakaryocytic differentiation of primary human CD34(+) cells cultured in the presence of thrombopoietin. The effect of MKL1 is abrogated when SRF is knocked down, suggesting that MKL1 acts through SRF. Consistent with these findings in human cells, knockout of Mkl1 in mice leads to reduced platelet counts in peripheral blood, and reduced ploidy in bone marrow megakaryocytes. In conclusion, MKL1 promotes physiologic maturation of human and murine megakaryocytes. PMID- 19136661 TI - T-cell responses associated with neonatal alloimmune thrombocytopenia: isolation of HPA-1a-specific, HLA-DRB3*0101-restricted CD4+ T cells. AB - T-cell responses have been implicated in the development of HPA-1a-induced neonatal alloimmune thrombocytopenia (NAIT). However, HPA-1a-specific T cells have neither been isolated nor characterized. Here, we aimed to determine whether HPA-1a-specific T cells could be isolated from HPA-1a-immunized women. In the present study, peripheral blood mononuclear cells (PBMCs) from an HPA-1a alloimmunized woman were cultured for weeks in the presence of HPA-1a peptide, labeled with CFSE, and assayed for antigen-specific proliferation. Individual proliferating cells were isolated by fluorescence-activated cell sorting and expanded in culture. Antigen specificity and HLA restriction were determined by cytokine secretion (enzyme-linked immunospot [ELISPOT]) and proliferation assays. Several CD3(+)CD4(+) T-cell clones were isolated that proliferated and secreted cytokines in response to HPA-1a peptide. Two of these clones have been established in long-term culture in our laboratory. Both of these recognize synthetic as well as naturally processed HPA-1a antigen, and the recognition is restricted by the MHC molecule HLA-DRB3*0101 that is strongly associated with NAIT. These HPA-1a-specific T-cell clones represent unambiguous evidence for the association of T-cell responses with NAIT, and they will serve as unique tools to elucidate the cellular immune response that may result in NAIT. PMID- 19136662 TI - Complement-dependent T-cell lymphopenia caused by thymocyte deletion of the membrane complement regulator Crry. AB - Although complement lysis is frequently used for the purification of lymphocyte subpopulations in vitro, how lymphocytes escape complement attack in vivo has not been clearly delineated. Here, we show that conditional gene targeting of a murine membrane complement regulator Crry on thymocytes led to complement dependent peripheral T-cell lymphopenia. Notably, despite evidence of hypersensitivity to complement attack, Crry-deficient T cells escaped complement injury and developed normally in the thymus, because of low intrathymic complement activity. Crry-deficient T cells were eliminated in the periphery by a C3- and macrophage-mediated but C5-independent mechanism. Thus, Crry is essential for mature T-cell survival in the periphery but not for lymphogenesis in the thymus. The observation that the thymus is a complement-privileged site may have implications for complement-based antitumor therapies. PMID- 19136663 TI - In silico analysis of phosphoproteome data suggests a rich-get-richer process of phosphosite accumulation over evolution. AB - Recent phosphoproteome analyses using mass spectrometry-based technologies have provided new insights into the extensive presence of protein phosphorylation in various species and have raised the interesting question of how this protein modification was gained evolutionarily on such a large scale. We investigated this issue by using human and mouse phosphoproteome data. We initially found that phosphoproteins followed a power-law distribution with regard to their number of phosphosites: most of the proteins included only a few phosphosites, but some included dozens of phosphosites. The power-law distribution, unlike more commonly observed distributions such as normal and log-normal distributions, is considered by the field of complex systems science to be produced by a specific rich-get richer process called preferential attachment growth. Therefore, we explored the factors that may have promoted the rich-get-richer process during phosphosite evolution. We conducted a bioinformatics analysis to evaluate the relationship of amino acid sequences of phosphoproteins with the positions of phosphosites and found an overconcentration of phosphosites in specific regions of protein surfaces and implications that in many phosphoproteins these clusters of phosphosites are activated simultaneously. Multiple phosphosites concentrated in limited spaces on phosphoprotein surfaces may therefore function biologically as cooperative modules that are resistant to selective pressures during phosphoprotein evolution. We therefore proposed a hypothetical model by which the modularization of multiple phosphosites has been resistant to natural selection and has driven the rich-get-richer process of the evolutionary growth of phosphosite numbers. PMID- 19136664 TI - Mitochondria do not contain lipid rafts, and lipid rafts do not contain mitochondrial proteins. AB - Lipid rafts are membrane microdomains involved in many cellular functions, including transduction of cellular signals and cell entry by pathogens. Lipid rafts can be enriched biochemically by extraction in a nonionic detergent at low temperature, followed by floatation on a sucrose density gradient. Previous proteomic studies of such detergent-resistant membranes (DRMs) are in disagreement about the presence of mitochondrial proteins in raft components. Here, we approach the status of mitochondrial proteins in DRM preparations by employing stable isotope labeling by amino acids in cell culture to evaluate the composition of differentially purified subcellular fractions as well as high resolution linear density gradients. Our data demonstrate that F(1)/F(0) ATPase subunits, voltage-dependent anion selective channels, and other mitochondrial proteins are at best partially copurifying contaminants of raft preparations. PMID- 19136665 TI - Increased susceptibility to diet-induced gallstones in liver fatty acid binding protein knockout mice. AB - Quantitative trait mapping identified a locus colocalizing with L-Fabp, encoding liver fatty acid binding protein, as a positional candidate for murine gallstone susceptibility. When fed a lithogenic diet (LD) for 2 weeks, L-Fabp(-/-) mice became hypercholesterolemic with increased hepatic VLDL cholesterol secretion. Seventy-five percent of L-Fabp(-/-) mice developed solid gallstones compared with 6% of wild-type mice with an increased gallstone score (3.29 versus 0.62, respectively; P < 0.01). Hepatic free cholesterol content, biliary cholesterol secretion, and the cholesterol saturation index of hepatic bile were increased in LD-fed L-Fabp(-/-) mice. Chow-fed L-Fabp(-/-) mice demonstrated increased fecal bile acid (BA) excretion accompanied by decreased ileal Asbt expression. By contrast, there was an increased BA pool and decreased fecal BA excretion in LD fed L-Fabp(-/-) mice, associated with increased proximal intestinal Asbt mRNA expression, suggesting that intestinal BA absorption was enhanced in LD-fed L Fabp(-/-) mice. The increase in biliary BA secretion and enterohepatic pool size in LD-fed L-Fabp(-/-) mice was accompanied by downregulation of Cyp7a1 mRNA and increased intestinal mRNA abundance of Fgf-15, Fxr, and Fabp6. These findings suggest that changes in hepatic cholesterol metabolism and biliary lipid secretion as well as changes in enterohepatic BA metabolism increase gallstone susceptibility in LD fed L-Fabp(-/-) mice. PMID- 19136666 TI - Phenolic acids suppress adipocyte lipolysis via activation of the nicotinic acid receptor GPR109A (HM74a/PUMA-G). AB - Phenolic acids are found in abundance throughout the plant kingdom. Consumption of wine or other rich sources of phenolic acids, such as the "Mediterranean diet," has been associated with a lower risk of cardiovascular disease. The underlying mechanism(s), however, has remained unclear. Here, we show that many phenolic acids, including those from the hydroxybenzoic and hydroxycinnamic acid classes, can bind and activate GPR109A (HM74a/PUMA-G), the receptor for the antidyslipidemic agent nicotinic acid. In keeping with this activity, treatment with a number of phenolic acids, including cinnamic acid, reduces lipolysis in cultured human adipocytes and in fat pats isolated from wild-type mice but not from mice deficient of GPR109A. Oral administration of cinnamic acid significantly reduces plasma levels of FFA in the wild type but not in mice deficient of GPR109A. Activation of GPR109A by phenolic acids may thus contribute to a cardiovascular benefit of these plant-derived products. PMID- 19136667 TI - Oxidized LDL impair adipocyte response to insulin by activating serine/threonine kinases. AB - Oxidized LDL (oxLDL) increase in patients affected by type-2 diabetes, obesity, and metabolic syndrome. Likewise, insulin resistance, an impaired responsiveness of target tissues to insulin, is associated with those pathological conditions. To investigate a possible causal relationship between oxLDL and the onset of insulin resistance, we evaluated the response to insulin of 3T3-L1 adipocytes treated with oxLDL. We observed that oxLDL inhibited glucose uptake (-40%) through reduced glucose transporter 4 (GLUT4) recruitment to the plasma membrane (-70%), without affecting GLUT4 gene expression. These findings were associated to the impairment of insulin signaling. Specifically, in oxLDL-treated cells insulin receptor (IR) substrate-1 (IRS-1) was highly degraded likely because of the enhanced Ser(307)phosphorylation. This process was largely mediated by the activation of the inhibitor of kappaB-kinase beta (IKKbeta) and the c-Jun NH(2) terminal kinase (JNK). Moreover, the activation of IKKbeta positively regulated the nuclear content of nuclear factor kappaB (NF-kappaB), by inactivating the inhibitor of NF-kappaB (IkappaBalpha). The activated NF-kappaB further impaired per se GLUT4 functionality. Specific inhibitors of IKKbeta, JNK, and NF-kappaB restored insulin sensitivity in adipocytes treated with oxLDL. These data provide the first evidence that oxLDL, by activating serine/threonine kinases, impaired adipocyte response to insulin affecting pathways involved in the recruitment of GLUT4 to plasma membranes (PM). This suggests that oxLDL might participate in the development of insulin resistance. PMID- 19136668 TI - Suberoylanilide hydroxamic acid reactivates HIV from latently infected cells. AB - Human immunodeficiency virus (HIV) persists in a latent form in infected individuals treated effectively with highly active antiretroviral therapy (HAART). In part, these latent proviruses account for the rebound in viral replication observed after treatment interruption. A major therapeutic challenge is to purge this reservoir. In this study, we demonstrate that suberoylanilide hydroxamic acid (SAHA) reactivates HIV from latency in chronically infected cell lines and primary cells. Indeed, P-TEFb, a critical transcription cofactor for HIV, is released and then recruited to the viral promoter upon stimulation with SAHA. The phosphatidylinositol 3-kinase/Akt pathway is involved in the initiation of these events. Using flow cytometry-based single cell analysis of protein phosphorylation, we demonstrate that SAHA activates this pathway in several subpopulations of T cells, including memory T cells that are the major viral reservoir in peripheral blood. Importantly, SAHA activates HIV replication in peripheral blood mononuclear cells from individuals treated effectively with HAART. Thus SAHA, which is a Food and Drug Administration-approved drug, might be considered to accelerate the decay of the latent reservoir in HAART-treated infected humans. PMID- 19136670 TI - Scavenger receptor BI-mediated selective uptake is required for the remodeling of high density lipoprotein by endothelial lipase. AB - Endothelial lipase (EL) is a negative regulator of high density lipoprotein (HDL) cholesterol plasma levels, and scavenger receptor BI (SR-BI) is involved in remodeling of HDL. The present study investigates the requirement of SR-BI for the effects of EL-mediated phospholipid hydrolysis on HDL metabolism in vivo. In vitro, selective uptake from EL-modified HDL was 129% higher than selective uptake from control HDL in SR-BI-overexpressing cells (p=0.01). In vivo overexpression of human EL by means of recombinant adenovirus decreased HDL plasma levels significantly (p<0.01). Fast protein liquid chromatography analysis and agarose gel electrophoresis revealed that EL expression resulted in the generation of small pre-beta HDL particles in wild-type mice, whereas in SR-BI-/- mice small HDL were preferentially removed. In kinetic experiments the fractional catabolic rate (FCR) of HDL cholesteryl ester increased by 110% (p<0.001), and the FCR of HDL apolipoproteins increased by 64% (p<0.001) in response to EL overexpression in wild-type mice. In SR-BI-/- mice a similar increase in the HDL apolipoprotein FCR occurred (p<0.001); however, there was no further increase in HDL cholesteryl ester catabolism. The apparent whole body selective uptake was increased 3-fold by EL in wild-type mice (p<0.001), whereas there was no selective uptake in SR-BI knock-out mice. EL overexpression increased hepatic selective uptake as well as holoparticle uptake (each p<0.01) in wild-type mice, whereas in SR-BI knock-out mice only holoparticle uptake increased (p<0.01). Our results indicate that SR-BI-mediated selective uptake of HDL cholesteryl ester is essential for the remodeling of large alpha-migrating HDL particles by EL. PMID- 19136669 TI - Growth factor regulation of growth factors in articular chondrocytes. AB - Several lines of evidence indicate that polypeptide growth factors are important in articular cartilage homeostasis and repair. It is not yet clear how these growth factors are regulated. We tested the hypothesis that the growth factors responsible for regulating cartilage are themselves regulated by growth factors. We delivered insulin-like growth factor I (IGF-I), fibroblast growth factor-2 (FGF-2), and/or transforming growth factor-beta1 (TGF-beta1) to adult bovine articular chondrocytes in primary culture and measured the resulting changes in IGF-I, FGF-2, and TGF-beta1 gene expression and protein production. These growth factors differentially regulated their own and each others gene expression and protein production. In concert, they regulated each other in an interactive fashion. Their interactions ranged from inhibitory to synergistic. The time course of the regulatory effects differed among the individual growth factors and combinations. Growth factor-induced changes in growth factor protein production by articular chondrocytes generally corresponded to the changes in gene expression patterns. These studies suggest that interactions among IGF-I, FGF-2, and TGF-beta1 substantially modulate their regulatory functions. The results may help guide the application of growth factors to articular cartilage repair. PMID- 19136673 TI - Anti-Mullerian hormone-based approach to controlled ovarian stimulation for assisted conception. AB - BACKGROUND: Individualization of controlled ovarian stimulation (COS) for assisted conception is complicated by variable ovarian response to follicle stimulating hormone. We hypothesized that anti-Mullerian hormone (AMH), a predictor of oocyte yield, may facilitate treatment strategies for women undergoing COS, to optimize safety and clinical pregnancy rates. METHODS: Prospective cohort study of 538 patients in two centres with differential COS strategies based on a centralized AMH measurement. RESULTS: AMH was associated with oocyte yield after ovarian stimulation in both centres, and a 'reduced' AMH (1 to <5 pmol/l) was associated with a reduced clinical pregnancy rate. Women with a 'normal' AMH (5 to <15 pmol/l) treated with a long GnRH-agonist protocol (both centres) showed a low incidence of excess response (0%) and poor response (0%). In women with 'high' AMH (>15 pmol/l), the antagonist protocol eliminated the need for complete cryopreservation of embryos due to excess response (P < 0.001) and showed a higher fresh cycle clinical pregnancy rate than agonist cycles [OR 4.40 (95% CI 1.95-9.93), P < 0.001]. CONCLUSIONS: The use of circulating AMH to individualize treatment strategies for COS may result in reduced clinical risk, optimized treatment burden and maintained pregnancy rates, and is worthy of prospective randomized examination. PMID- 19136672 TI - Type XIV Collagen Regulates Fibrillogenesis: PREMATURE COLLAGEN FIBRIL GROWTH AND TISSUE DYSFUNCTION IN NULL MICE. AB - Type XIV collagen is a fibril-associated collagen with an interrupted triple helix. This collagen interacts with the fibril surface and has been implicated as a regulator of fibrillogenesis; however, a specific role has not been elucidated. Functional roles for type XIV collagen were defined utilizing a new type XIV collagen-deficient mouse line. This line was produced using a conventional targeted knock-out approach. Col14a1(-/-) mice were devoid of type XIV collagen, whereas heterozygous mice had reduced synthesis. Both mutant Col14a1 genotypes were viable with a grossly normal phenotype; however, mature skin exhibited altered mechanical properties. Prior to evaluating tendon fibrillogenesis in type XIV collagen-deficient mice, the developmental expression patterns were analyzed in wild-type flexor digitorum longus (FDL) tendons. Analyses of mRNA and protein expression indicated tissue-specific temporal expression that was associated with the early stages in fibrillogenesis. Ultrastructural analyses of wild-type and null tendons demonstrated premature fibril growth and larger fibril diameters in tendons from null mice at postnatal day 4 (P4). However, fibril structure in mature tendons was normal. Biomechanical studies established a direct structure/function relationship with reduced strength in P7-null tendons. However, the biomechanical properties in P60 tendons were comparable in null and wild-type mice. Our results indicate a regulatory function for type XIV collagen in early stages of collagen fibrillogenesis with tissue differences. PMID- 19136671 TI - Genetic evidence for the coordinated regulation of collagen fibrillogenesis in the cornea by decorin and biglycan. AB - Decorin and biglycan are class I small leucine-rich proteoglycans (SLRPs) involved in regulation of collagen fibril and matrix assembly. We hypothesize that tissue-specific matrix assembly, such as in the cornea, requires a coordinate regulation involving multiple SLRPs. To this end, we investigated the expression of decorin and biglycan in the cornea of mice deficient in either SLRP gene and in double-mutant mice. Decorin and biglycan exhibited overlapping spatial expression patterns throughout the corneal stroma with differential temporal expression. Whereas decorin was expressed at relatively high levels in all developmental stages, biglycan expression was high early, decreased during development, and was present at very low levels in the mature cornea. Ultrastructural analyses demonstrated comparable fibril structure in the decorin- and biglycan-null corneas compared with wild-type controls. We found a compensatory up-regulation of biglycan gene expression in the decorin-deficient mice, but not the reverse. Notably, the corneas of compound decorin/biglycan-null mice showed severe disruption in fibril structure and organization, especially affecting the posterior corneal regions, corroborating the idea that biglycan compensates for the loss of decorin. Fibrillogenesis assays using recombinant decorin and biglycan confirmed a functional compensation, with both having similar effects at high SLRP/collagen ratios. However, at low ratios decorin was a more efficient regulator. The use of proteoglycan or protein core yielded comparable results. These findings provide firm genetic evidence for an interaction of decorin and biglycan during corneal development and further suggest that decorin has a primary role in regulating fibril assembly, a function that can be fine-tuned by biglycan during early development. PMID- 19136674 TI - Mutations at serine 37 in mouse guanylate kinase confer resistance to 6 thioguanine. AB - Guanylate kinase (GMK) is an essential nucleoside monophosphate kinase that catalyzes the phosphorylation of guanine-monophosphate (GMP) and dGMP to yield GDP and dGDP, respectively, important precursors for nucleotide synthesis. GMK is also responsible for the activation of 6-thioguanine (6-TG), a drug widely used as chemotherapeutic agent to treat leukemia. Several mechanisms of resistance to 6-TG have been reported but a subset of drug resistant cells cannot be explained by these mechanisms. We propose that mutations in GMK could result in drug resistance. Because cells require the presence of a functional GMK for viability, mutations that arise that lead to 6-TG resistance must retain activity toward GMP. We report three amino acid substitutions at serine 37 (S37) in mouse GMK that display activity toward GMP by conferring genetic complementation to a conditional GMK-deficient Escherichia coli and in enzyme assays. When 6-TG is included in complementation studies, cells expressing wild-type GMK are sensitive whereas all S37 mutants examined are able to effectively discriminate against 6 TG and display a drug resistance phenotype. Activity of the three S37 mutant enzymes toward clinically relevant concentrations of 6-TGMP is undetectable. Mutations in GMK, therefore, represent a previously undescribed mechanism for 6 TG resistance. PMID- 19136675 TI - The influence of the framework core residues on the biophysical properties of immunoglobulin heavy chain variable domains. AB - Antibody variable domains differ considerably in stability. Single-chain Fv (scFv) fragments derived from natural repertoires frequently lack the high stability needed for therapeutic application, necessitating reengineering not only to humanize their sequence, but also to improve their biophysical properties. The human V(H)3 domain has been identified as having the best biophysical properties among human subtypes. However, complementarity determining region (CDR) grafts from highly divergent V(H) domains to huV(H)3 frequently fail to reach its superior stability. In previous experiments involving a CDR graft from a murine V(H)9 domain of very poor stability to huV(H)3, a hybrid V(H) framework was obtained which combines the lower core residues of muV(H)9 with the surface residues of huV(H)3. It resulted in a scFv with far better biophysical properties than the corresponding grafts to the consensus huV(H)3 framework. To better understand the origin of the superior properties of the hybrid framework, we constructed further hybrids, but now in the context of the consensus CDR-H1 and -H2 of the original human V(H)3 domain. The new hybrids included elements from either murine V(H)9, human V(H)1 or human V(H)5 domains. From guanidinium chloride-induced equilibrium denaturation measurements, kinetic denaturation experiments, measurements of heat-induced aggregation and comparison of soluble expression yield in Escherichia coli, we conclude that the optimal V(H) framework is CDR-dependent. The present work pinpoints structural features responsible for this dependency and helps to explain why the immune system uses more than one framework with different structural subtypes in framework 1 to optimally support widely different CDRs. PMID- 19136677 TI - Impact of health education on health-related quality of life among elderly persons: results from a community-based intervention study in rural Bangladesh. AB - This study examines the change in health-related quality of life (HRQoL) among (> or =60 years) elderly persons as a result of health education intervention. A community-based intervention study was performed in eight randomly selected villages ( INTERVENTION: n = 4; CONTROL: n = 4) in rural Bangladesh. A total of 1135 elderly persons was selected for this study. The analyses include 839 participants ( INTERVENTION: n = 425; CONTROL: n = 414) who participated in both baseline and post-intervention surveys. Participants in the intervention area were further stratified into compliant (n = 315) and non-compliant (n = 110) groups based on the reported compliance to the intervention activities. The intervention includes, for example, physical activity, advice on healthy food intake and other aspects of management. To create an enabling environment, social awareness was provided by means of information about the contribution of and challenges faced by elderly persons at home and the community, including information about elderly persons' health and health care. The intervention activities were provided to the elderly persons, caregivers, household members and community people for 15 months. The HRQoL was assessed using a multi dimensional generic instrument designed for elderly persons. Multivariate analyses revealed that in the non-compliant group the probabilities of increased scores were less likely in overall HRQoL (OR 0.52, 95% CI 0.32-0.82). Among the CONTROL group, increased scores were less likely in the physical (OR 73, 95% CI 0.54-0.99), social (OR 0.37, 95% CI 0.27-0.50), spiritual (OR 0.60, 95% CI 0.34 0.94), environment (OR 0.36, 95% CI 0.26-0.49) dimensions and overall HRQoL (OR 0.44, 95% CI 0.32-0.59) (adjusted for age, sex, literacy, marital status and economic status). This study concludes that provision of community-based health education intervention might be a potential public health initiative to enhance the HRQoL in old age. PMID- 19136676 TI - Health promotion profile of youth sports clubs in Finland: club officials' and coaches' perceptions. AB - The purpose of this article is to examine the current health promotion orientation of youth sports clubs in Finland in view of the standards created previously for the health promoting sports club (HPSC). Ninety-seven youth sports clubs participated, and 273 sports club officials and 240 coaches answered the questionnaires. To describe clubs health promotion orientations, an HPSC index was created. The HPSC index was formulated on sub-indices by factor analysis. The sub-indices were: policy, ideology, practice and environment indexes. The results indicate that youth sports clubs are fairly health promoting in general. On average, the clubs fulfilled 12 standards for HPSC out of 22. Every fourth club was categorized as higher health promoting (> or = 15 fulfilled standards), and every third as lower health promoting (<11 fulfilled standards). The variation between clubs was wide. The clubs that had been recognized as exemplary and hence certified by the Young Finland Association were more likely to recognize health promotion than non-certified clubs (OR = 2.36, p = 0.016). The sports club officials were twice as likely to evaluate their clubs as higher health promoting than the coaches (OR = 2.04, p = 0.041). Under the sub-indices, ideologies were recognized best, others less. These findings indicate that minority of the youth sports clubs have realized health promotion comprehensively as a part of their activities. There is a lot of need for development, especially in the area of health promotion policies and practices. The instruments used proved valid and reliable and can therefore be recommended for international use. PMID- 19136678 TI - Discordant genotypic interpretation and phenotypic role of protease mutations in HIV-1 subtypes B and G. AB - OBJECTIVES: HIV-1 group M is classified into nine different subtypes. Most antiretroviral (ARV) drugs have been developed for subtype B, and the response of non-B subtypes in terms of susceptibility and the acquisition of drug resistance when facing those drugs is largely unknown. In this study, we aimed to address differences in the impact of protease inhibitor (PI)-selected mutations on subtypes B and G. PATIENTS AND METHODS: ARV-treated, HIV-positive patients regularly monitored at the Hospital de Egas Moniz, in Lisbon, Portugal, were examined for the presence of PI-associated primary mutations (301 subtype B and 184 subtype G), and for the selection of those mutations over the time of PI exposure. Forty-three subtype G patients were phenotyped for susceptibility to all available PIs through VIRCO's Antivirogram, and compared with a similar dataset of subtype B patients. RESULTS: Mutation I54V/L was selected by nelfinavir in subtype G isolates, a mutation not previously described for this drug in subtype B. L90M was associated with a lower reduction in the susceptibility of subtype G to nelfinavir when compared with subtype B, and with no reduced susceptibility to saquinavir. This was compensated for by the acquisition of M89I in subtype G. L90M did not reduce the susceptibility of subtype G to saquinavir, in contrast to subtype B. Likewise, the pattern I54V/L L90M did not reduce the susceptibility of subtype G to indinavir and saquinavir. Indinavir-associated mutations M46I/L, I84V and V82A/F/T developed earlier in subtype B across the time of exposure to that drug when compared with subtype G counterparts. CONCLUSIONS: Our results provide proof of principle and support the growing evidence that subtype-specific responses to ARVs exist. Data presented here highlight inconsistencies in current genotyping interpretation algorithms inadequately applied to all HIV-1 subtypes. PMID- 19136679 TI - Comparison of three phenotypic techniques for detection of methicillin resistance in Staphylococcus spp. reveals a species-dependent performance. AB - OBJECTIVES: To evaluate the usefulness of the cefoxitin screen in Vitek 2 Gram positive panels for recognizing methicillin-resistant strains of staphylococci. METHODS: Seven hundred and ninety-nine non-duplicate isolates of Staphylococcus aureus and coagulase-negative strains were included in the study. Methicillin resistance was measured using PCR for the mecA gene, the CLSI cefoxitin disc diffusion method, the Vitek 2 cefoxitin screen and the Vitek 2 oxacillin susceptibility test. RESULTS: Compared with the molecular detection of methicillin resistance the overall sensitivities and specificities of the phenotypic tests for cefoxitin disc diffusion were 94.9% and 97.0%, for Vitek 2 cefoxitin screen were 94.6% and 93.5% and for Vitek 2 oxacillin susceptibility test were 93.8% and 77.9%. The cephamycin tests (cefoxitin disc diffusion and Vitek 2 screen) were not able to identify mecA-positive strains of Staphylococcus simulans. In addition, the performance of the Vitek 2 system was poor against Staphylococcus cohnii subspecies, Staphylococcus hominis hominis and Staphylococcus saprophyticus. CONCLUSIONS: Overall, the performance of the Vitek 2 system for differentiating mecA-positive staphylococci was comparable to PCR and the CLSI disc diffusion method; however, performance was species-dependent. Thus, before accepting the results produced by Vitek 2, species identification may be required. PMID- 19136680 TI - Physical exertion and triggering of myocardial infarction. PMID- 19136681 TI - A novel functional haplotype in the human GNAS gene alters Galphas expression, responsiveness to beta-adrenoceptor stimulation, and peri-operative cardiac performance. AB - AIMS: Cardiac overexpression of the beta-adrenoceptor-coupled G-protein subunit Galphas in mice enhances inotropic responses to sympathetic stimulation, but evokes cardiomyopathy with increasing age. We tested whether functional single nucleotide polymorphisms (SNPs) in the human Galphas (GNAS) gene modulate Galphas expression and assessed functional consequences. METHODS AND RESULTS: Sequencing the promoter and intron 1 of GNAS revealed 11 SNPs resulting in three common haplotypes. Haplotype *3 constructs exhibited significantly higher promoter activity than haplotypes *1 and *2, resulting in a more than 50% higher Galphas mRNA expression in homozygous *3 carriers (*3/*3) than in heterozygous (*3/-) and negative *3 (-/-) carriers (P = 0.002). Basal, Galphas- (via NaF and GTP) and isoproterenol-stimulated adenylyl cyclase (AC) activities were also significantly higher in *3/*3 than in *3/- and -/- carriers. In contrast, direct AC activation via forskolin was independent of GNAS haplotypes. Furthermore, haemodynamic measurements in 137 coronary artery bypass patients revealed a higher cardiac index in *3/*3 carriers than in *3/- and -/- carriers (P = 0.025) associated with a lower NYHA functional class (P = 0.040) and serum NT-proBNP concentrations (P = 0.002). CONCLUSION: SNPs in regulatory regions of GNAS impact upon Galphas expression and stimulated cAMP formation in human hearts in vitro and upon cardiac performance in vivo. PMID- 19136683 TI - Prevalence of thermophilic Campylobacter species in household cats and dogs in Ireland. AB - Rectal swabs were collected from 147 household dogs and 35 household cats, including healthy animals, animals with gastrointestinal signs and animals with a variety of medical and surgical conditions. A combination of selective culture methods was used to optimise the recovery of Campylobacter species, and a PCR was used to confirm their isolation and to identify the species. The overall prevalence of Campylobacter species was 42.9 per cent in the cats and 41.5 per cent in the dogs. Campylobacter upsaliensis was the species most commonly isolated from the dogs and cats, and Campylobacter jejuni was the second most commonly isolated. Particularly high prevalences were detected in the few cats and dogs with diarrhoea, and in the cats and dogs that were six months old or younger. PMID- 19136684 TI - Effects of a modified live CSFV vaccine on the development of PMWS in pigs infected experimentally with PCV-2. AB - The objective of this study was to determine the effect of vaccination against classical swine fever virus (CSFV) on the development of postweaning multisystemic wasting syndrome (PMWS) in conventional pigs infected experimentally with porcine circovirus type 2 (PCV-2). The pigs infected with PCV 2 and immunised with modified live CSFV developed mild to moderate PMWS, whereas none of the pigs infected with PCV-2 alone or immunised with modified live CSFV alone developed PMWS. Lesions histologically characteristic of PMWS were observed in lymph nodes from the pigs infected with PCV-2 and immunised with modified live CSFV vaccine, and extensive replication of PCV-2 was detected in the nodes by in situ hybridisation. PMID- 19136685 TI - Assessing techniques for disinfecting sites for inserting intravenous catheters into the jugular veins of horses. AB - The sites of insertion of catheters into the jugular veins of six horses were investigated to determine common isolates and to assess the effectiveness of two disinfection protocols with the hair coat left long, clipped or shaved. Skin commensals (Staphylococcus, Streptococcus and Micrococcus species) and environmental contaminants (Bacillus, Enterobacteriaceae, Aspergillus and Mucor species) were the microorganisms most frequently isolated. Chlorhexidine gluconate and povidone-iodine-based skin disinfection protocols resulted in significant reductions in the number of bacterial isolates from clipped sites. With chlorhexidine, there were no significant differences between the reductions observed at sites with the hair coat left long, clipped or shaved. PMID- 19136686 TI - Comparative evaluation of diagnostic techniques for bovine viral diarrhoea virus in aborted and stillborn fetuses. PMID- 19136687 TI - Bartonella clarridgeiae in a cat in the UK. PMID- 19136688 TI - Prevalence of Rhipicephalus sanguineus ticks on dogs in a region on the Mexico USA border. PMID- 19136689 TI - Haemophilia in Maine Coon cats. PMID- 19136690 TI - Unusual bovine papular stomatitis virus infection in a British dairy cow. PMID- 19136691 TI - Treatment of pain and inflammation in degenerative joint disease. PMID- 19136692 TI - Capillaria hepatica in primates in zoological collections in the British Isles. PMID- 19136693 TI - The role of twitching motility in Pseudomonas aeruginosa exit from and translocation of corneal epithelial cells. AB - PURPOSE: The authors have shown that twitching motility, a pilus-mediated form of bacterial surface movement, is required for Pseudomonas aeruginosa virulence in a murine model of keratitis. To study the role of twitching motility in virulence, Pseudomonas traversal of multilayered corneal epithelia in vitro was investigated. METHODS: Translocation of multilayered corneal epithelia was investigated with the invasive strain PAK and isogenic twitching motility mutants. Rabbit corneal epithelial cells were grown to multilayers with filters and inoculated on their apical surfaces with 10(6) colony-forming unit bacteria, and translocating bacteria were quantified by viable counts of the basal chamber. Transepithelial resistance (TER) was recorded. Cellular exit of P. aeruginosa after invasion was quantified with modified gentamicin survival assays, and the role of apoptosis in exit was explored. RESULTS: PAK translocated the epithelia as early as 1 hour after infection, and by 8 hours apical and basal numbers of bacteria were similar. Bacterial translocation did not reduce TER. Each twitching motility mutant (pilU, pilT with pili, pilA lacking pili) was defective in translocation (>2 log reduction vs. PAK; P < 0.005). All twitching mutants were competent for cell invasion but defective in cellular exit, accumulating intracellularly to numbers exceeding those of PAK. Inhibiting apoptosis reduced the cellular exit of PAK. CONCLUSIONS: These results show that twitching motility enables P. aeruginosa to translocate corneal epithelial layers and suggest that it contributes to epithelial cell exit by a mechanism involving apoptosis. The relationship between these in vitro findings and the role of twitching motility in P. aeruginosa virulence in vivo remains to be determined. PMID- 19136694 TI - Comparison of clinical and three-dimensional histomorphometric optic disc margin anatomy. AB - PURPOSE: To investigate the anatomic basis of the optic disc margin in the normal monkey eye by colocalizing optic disc photographs to three-dimensional (3D) histomorphometric reconstructions of the same optic nerve head. METHODS: Optic disc photographs from 28 normal monkey eyes were overlaid onto 3D central retinal vessel reconstructions generated as part of postmortem optic nerve histomorphometric reconstructions for each eye. Within each reconstruction, the Bruch's membrane opening (BMO) was delineated. Alignment was achieved by matching the clinical vessel outline to the vessel reconstruction with parallel viewing software. An experienced observer viewed stereophotographs and marked the disc margin onto clinical photographs with custom software. Alignment of the delineated disc margin to the histomorphometrically defined BMO was qualitatively assessed within each image. RESULTS: In 20 eyes, BMO aligned well to the disc margin delineation. In four eyes, alignment improved after repeated colocalization. Careful review of the histomorphometric reconstructions identified that in most cases Bruch's membrane extended beyond the termination of the border tissue of Elschnig, most substantially in the superior and nasal sectors. Misalignments could be explained by inaccurate BMO marking or where Bruch's membrane terminated externally to the inferior edge of the border tissue; this latter structure aligned to the disc margin. CONCLUSIONS: BMO was a clinically detectable entity and represented the disc margin in most eyes in this study. The 3D architecture of the border tissue combined with the presence of an overhang of Bruch's membrane makes an important contribution to disc margin anatomy. PMID- 19136695 TI - Chemical inhibition of alpha-toxin, a key corneal virulence factor of Staphylococcus aureus. AB - PURPOSE: alpha-Toxin mediates extreme corneal damage during Staphylococcus aureus keratitis. Chemical inhibition of this toxin was sought to provide relief from toxin-mediated pathology. METHODS: Inhibition of alpha-toxin by phosphate buffered saline (PBS), 0.1% methyl-beta-cyclodextrin (CD), or CD plus cholesterol (0.1%, CD-cholesterol) was assayed by hemolysis of rabbit erythrocytes. Pathologic changes in rabbit corneas injected with 12 hemolytic units of alpha toxin suspended in PBS, 1% CD, or 1% CD-cholesterol were compared over time. Rabbit corneas injected with 10(2) colony forming units (CFU) of S. aureus were treated from 7 to 13 hours postinfection (PI) with a total of 15 drops of CD cholesterol, CD, or PBS. Slit lamp examination (SLE) and measurement of erosions were performed at 13 hours PI and bacteria were quantified at 14 hours PI. RESULTS: Toxin-mediated lysis of erythrocytes was inhibited up to 16,000-fold in the presence of CD-cholesterol compared with CD or PBS. Eyes injected with alpha toxin mixed with CD-cholesterol had, at 7 hours postinjection, significantly smaller erosions than eyes injected with alpha-toxin in PBS or alpha-toxin mixed with CD (P = 0.0090 and P = 0.0035, respectively). Eyes infected with S. aureus and treated with CD-cholesterol had significantly lower SLE scores than eyes treated with CD or PBS (P or= 0.0648). CONCLUSIONS: CD-cholesterol is a potent inhibitor of alpha-toxin activity in vitro and an effective means to arrest corneal damage during S. aureus keratitis. PMID- 19136696 TI - Photoreceptors repair by autologous transplantation of retinal pigment epithelium and partial-thickness choroid graft in rabbits. AB - PURPOSE: To investigate whether autologous retinal pigment epithelium (RPE) and a partial-thickness graft can repair degenerated photoreceptors overlying a mechanically damaged Bruch's membrane. METHODS: Twenty-one pigmented rabbits were used in the study. Abrasive debridement of the RPE was performed with a metal cannula after superior retinal bleb detachment in 20 rabbits. The graft was prepared beneath the inferior retina and was transplanted to the debridement area 14 days later. Debridement-only sites served as the control. Tissue sections were evaluated by light microscopy and transmission electron microscopy at 7 days, 1 month, and 3 months after transplantation, corresponding to 21 days, 45 days, and 3 months after debridement, respectively. RESULTS: When analyzed at 7 days after transplantation, short buds of inner segment with regularly organized outer nuclear layer were observed. The outer segments (OS) were of insufficient length to be observed, but by 1 and 3 months, a significant elongation of the OS was detected. In control retinas from 21 days (corresponding to 7 days after transplantation) to 3 months after RPE debridement, the outer nuclear layer cells were disorganized and diminished. CONCLUSIONS: This study showed that autologous RPE and partial-thickness choroid graft have the capacity not only to support photoreceptor cell survival, but also to initiate early repair mechanisms, as exhibited by outer segment regeneration. PMID- 19136697 TI - Utility of molecular sequence analysis of the ITS rRNA region for identification of Fusarium spp. from ocular sources. AB - PURPOSE: Fungal ocular infections cause significant ocular morbidity, particularly when diagnosis and treatment are delayed. Accurate morphologic identification of Fusarium spp. beyond the genus is time-consuming and insensitive. It was the purpose of this study to examine the usefulness of the nuclear ribosomal RNA (rRNA) internal transcribed spacer regions (ITS1 and -2) to detect and differentiate Fusarium spp. responsible for ocular infections. METHODS: Fifty-eight archived isolates from ocular sources of 52 patients diagnosed with Fusarium keratitis at the Bascom Palmer Eye Institute (Miami, FL) from April 2000 to May 2007 were analyzed. The archived samples, which were initially classified according to morphologic characteristics, were analyzed by DNA sequence data generated from the ITS regions of the rRNA genes. RESULTS: Fifteen distinct sequences were identified among the 58 isolates. Sequence analysis identified the isolates as Fusarium solani (75%), F. oxysporum (16%), F. incarnatum-equiseti (5%), F. dimerum (2%), and one Fusarium sp. (2%) that was not classified within any species complex. Species identification based on sequence data correlated well with the morphologic classification when performed by a mycology reference laboratory, but a higher rate of mismatch was observed based on identification by a nonreference laboratory. CONCLUSIONS: Most of the isolates of Fusarium ocular infections belong to the F. solani or F. oxysporum species complexes. Morphologic classification at the species level yielded inconsistent results at a general microbiology laboratory. In contrast, the sequence variation within the ITS region allowed reliable and faster discrimination of the isolates at both the genus and species level. PMID- 19136699 TI - MHC-matched corneal allograft rejection in an IFN-gamma/IL-17-independent manner in C57BL/6 mice. AB - PURPOSE: It has been widely accepted that Th1- and IFN-gamma-mediated immune responses are indispensable for corneal allograft rejection in BALB/c hosts. The present study was designed to determine the role of IFN-gamma and IL-17 in the rejection by C57BL/6 hosts, which display high rejection rates. METHODS: MHC matched or -mismatched corneal allografts were grafted onto IFN-gamma-knockout (GKO), IFN-gamma-receptor-knockout (GRKO), IL-17-knockout (IL-17KO), or wild-type (WT) C57BL/6 hosts. Graft fates were assessed clinically and histologically. At appropriate time intervals after allografting, RNA was isolated from corneal graft parenchymal and stromal tissues and cervical lymph nodes. The cytokine mRNA levels of Th1, -2, and -17 type were analyzed by real-time PCR. RESULTS: No significantly prolonged allograft survival was observed in any combinations. The rejected MHC-mismatched corneas in GKO elicited intensive infiltration of eosinophils, CD11b(+) macrophages, and B cells, but few Gr-1(+)CD11c(-) neutrophils. In contrast, rejected MHC-matched corneas in GKO hosts, as well as GRKO and WT hosts, elicited intensive infiltration of CD11b(+) macrophages and Gr 1(+)CD11c(-) neutrophils, but no B220(+) B cells and eosinophils. At 1 week after MHC-matched allografting, mRNA levels of IL-6 and IL-17A in the lymph node were extensively upregulated in GKO hosts. It is of interest that anti-IFN-gamma treatment did not improve the allograft survival in IL-17KO hosts. CONCLUSIONS: IFN-gamma and IL-17 play no critical role in the development of minor-specific allograft rejection in C57BL/6 mice. This indicates the presence of sophisticated rejection mechanisms that are still elusive and cannot be ascribed simply to Th1, -2, or -17. PMID- 19136698 TI - Prevention of endotoxin-induced uveitis in rats by benfotiamine, a lipophilic analogue of vitamin B1. AB - PURPOSE: To study the amelioration of ocular inflammation in endotoxin-induced uveitis (EIU) in rats by benfotiamine, a lipid-soluble analogue of thiamine. METHODS: EIU in Lewis rats was induced by subcutaneous injection of lipopolysaccharide (LPS) followed by treatment with benfotiamine. The rats were killed 3 or 24 hours after LPS injection, eyes were enucleated, aqueous humor (AqH) was collected, and the number of infiltrating cells, protein concentration, and inflammatory marker levels were determined. Immunohistochemical analysis of eye sections was performed to determine the expression of inducible-nitric oxide synthase (iNOS), cyclooxygenase (Cox)-2, protein kinase C (PKC), and transcription factor NF-kappaB. RESULTS: Infiltrating leukocytes, protein concentrations, and inflammatory cytokines and chemokines were significantly elevated in the AqH of EIU rats compared with control rats, and benfotiamine treatment suppressed these increases. Similarly increased expression of inflammatory markers iNOS and Cox-2 in ciliary body and retinal wall was also significantly inhibited by benfotiamine. The increased phosphorylation of PKC and the activation of NF-kappaB in the ciliary body and in the retinal wall of EIU rat eyes were suppressed by benfotiamine. CONCLUSIONS: These results suggest that benfotiamine suppresses oxidative stress-induced NF-kappaB-dependent inflammatory signaling leading to uveitis. Therefore, benfotiamine could be used as a novel therapeutic agent for the treatment of ocular inflammation, especially uveitis. PMID- 19136700 TI - The effects of 5-fluorouracil on ocular tissues in vitro and in vivo after controlled release from a multifunctional implant. AB - PURPOSE: To evaluate the effects of 5-fluorouracil (5-FU) on ocular cells in vitro and the effects of degradable 5-FU-loaded poly(DL-lactide-co-glycolide; PDLGA) 50:50 implant in the rabbit eye in vivo. METHODS: Cytotoxicity was assessed with a tetrazolium salt WST-1 cell proliferation/viability test and a lactate dehydrogenase (LDH) leakage test in rabbit corneal stromal fibroblasts (SIRCs), bovine corneal endothelial cells (BCECs), human conjunctival epithelial cells (IOBA-NHCs), human retinal pigment epithelial cells (ARPE-19), and human corneal epithelial cells (HCECs). The 5-FU-loaded PDLGA implants were surgically placed in rabbit eyes with a deep sclerectomy technique and the histopathology of the eyes was examined. RESULTS: In vitro, 5-FU affected cell proliferation and survival in a time- and dose-dependent manner. In the WST-1 test, adverse effects in serum-free conditions started from 0.0005 mg/mL 5-FU in SIRCS and HCECs, whereas in other cell types, 0.005 mg/mL 5-FU hindered cell proliferation. In serum-free conditions 72-hour 5 mg/mL 5-FU treatment decreased cell viability to 40% in BCECs and to 10% to 15% in other cell types. 5-FU had no or very minor effects on LDH leakage. In vivo, the 5-FU implant showed no signs of toxicity in cornea and retina, whereas in the conjunctival stroma near the implantation site, some inflammatory cells and a marked subepithelial condensation of stromal connective tissue was observed during the postoperative period of 4 weeks. CONCLUSIONS: 5-FU had a broad therapeutic range, and the 5-FU implant showed only minor tissue reactions in conjunctiva at the surgical site. 5-FU is a possible candidate for controlled drug release. PMID- 19136701 TI - Experimental glutamatergic excitotoxicity in rabbit retinal ganglion cells: block by memantine. AB - PURPOSE: Excessive activity of NMDA-type (N-methyl-d-aspartate) glutamatergic channels has been implicated as a mechanism for neuronal injury in neurologic disorders, including glaucoma, and retinal disease. This study was designed to characterize the retinal response to experimental manipulations that mimic features of glutamatergic excitotoxic insult and also to determine whether memantine, an NMDA-type glutamatergic channel blocker, is effective in reversing experimental excitotoxicity. METHODS: Recordings of the electroretinogram (ERG) and spiking activity of single retinal ganglion cells (RGCs) were made from rabbit retinas. Excitotoxic insult was induced by either (1) application of NMDA, a selective NMDA receptor agonist; (2) application of TBOA (dl-threo-beta benzyloxyaspartic acid), a selective inhibitor of glutamate transporters, or (3) perfusion with magnesium-free medium. For each condition, memantine was coapplied to determine its efficacy for reversal of experimental excitotoxicity. Memantine was also applied in isolation to characterize any effect on retinal responses to light stimuli. RESULTS: All three experimental manipulations were associated with an increase in the tonic level of RGC spiking activity, a reduction in RGC spike amplitude, and, in some cells, block of spike generation. Experimental excitotoxicity had little or no effect on ERG responses. Coapplication of memantine was associated with recovery of RGC tonic spiking activity and spike amplitude toward control levels. Application of memantine in isolation was associated with a dose-dependent effect on the timing of ERG and RGC-OFF responses. CONCLUSIONS: Memantine was effective in reversing acute experimental excitotoxicity at concentrations that have little effect on retinal light signaling. PMID- 19136702 TI - The regulation of NADPH oxidase and its association with cell proliferation in human lens epithelial cells. AB - PURPOSE: NADPH oxidase (NOX)-generated reactive oxygen species (ROS) are essential for growth factor-stimulated cell proliferation. In this study, the regulatory role of p22phox, a membrane subunit of NOX, in NOX activity and platelet-derived growth factor (PDGF) mitogenic signaling were examined. METHODS: Human lens epithelial B3 (HLE B3) cell lines with p22phox overexpressed (p22-OE) and p22phox knockdown (p22-KD) were used as models. Cells stimulated with PDGF were compared with nonstimulated control cells. The relative NOX activity and intracellular ROS generation were detected by lucigenin-based assay and DCFH fluorescence, respectively. Cell proliferation was measured by BrdU and fluorescent nucleic acid staining assays. p22phox, P-JNK, P-ERK1/2, P-Akt, P-p38, p47phox, and P-PDGF receptor in cell lysates were detected by Western blot analysis with the respective specific antibodies. RESULTS: p22-OE showed higher NOX activity, PDGF-stimulated ROS generation, cell proliferation, and activation of signaling cascades of ERK1/2, JNK, and Akt over the control (vector alone). In contrast, p22-KD displayed opposite results. In addition, PDGF stimulated p47phox and Rac1 translocations and induced binding between p22phox and the cytosolic subunits of p47phox, p67phox, and p40phox. Overexpression of p22phox increased p22phox-p47phox binding, enhanced, and prolonged the phosphorylation of PDGF receptor at Tyr857 with a corresponding inhibition of the activity of the oxidation-sensitive low molecular weight protein tyrosine phosphatase (LMW-PTP). However, p22phox knockdown weakened p22phox-p47phox binding and largely diminished the activation of PDGF receptor with no inhibition of LMW-PTP. CONCLUSIONS: PDGF mitogenic action in HLE B3 cells depends on p22phox to regulate NOX activity, which affects PDGF receptor function for cell proliferation. PMID- 19136703 TI - The use of human mesenchymal stem cell-derived feeder cells for the cultivation of transplantable epithelial sheets. AB - PURPOSE: To report the efficacy of human bone marrow-derived mesenchymal stem cells as a source of feeder cells for the cultivation of transplantable corneal epithelial cell sheets. METHODS: Human mesenchymal stem cells (marrow adherent stem cells; MASCs) were cultured in alpha-modified Eagle's medium with 10% serum and were treated with mitomycin C. Expression of cytokines in MASCs was confirmed by reverse transcription-polymerase chain reaction. Human limbal epithelial cells were cocultured with MASCs or 3T3 feeder cells to compare colony-forming efficiency (CFE). Limbal epithelial cells were cultured on MASCs or 3T3 feeder cells at the air-liquid interface to allow stratification, and stratified epithelial sheets were analyzed by immunohistochemistry against cytokeratin 3 (K3), K15, p63alpha, and ABCG2. Rabbit limbal epithelial cell sheets were cultivated with MASC feeder cells and transplanted to the ocular surface of the limbal-deficient rabbits. Epithelial grafts were observed by slit lamp microscopy for 4 weeks and then evaluated by histology and immunohistochemistry against K3 and K4. RESULTS: MASC feeder cells expressed keratinocyte growth factor, hepatocyte growth factor, and N-cadherin. The CFE of human limbal epithelial cells was similar in MASC and 3T3 feeder groups. Stratified cell sheets were successfully cultivated with MASC feeder cells expressing K3, K15, p63alpha, and ABCG2. Transplanted epithelial sheets regenerated the corneal phenotype in limbal deficient rabbits. CONCLUSIONS: MASC-derived feeder cells are suitable for the engineering of epithelial sheets, avoiding the use of potentially hazardous xenologic feeder cells. PMID- 19136704 TI - Hyperexpression of the high-affinity IgE receptor-beta chain in chronic allergic keratoconjunctivitis. AB - PURPOSE: Although the existence of Fc(epsilon)RI-alphabetagamma(2) and Fc(epsilon)RI-alphagamma(2) receptor subtypes was reported, there has been no direct evidence of these two subtypes of Fc(epsilon)RI in vivo. To investigate the existence of these two subtypes of Fc(epsilon)RI in vivo, the authors evaluated the expression of Fc(epsilon)RI-beta in the giant papillae of chronic allergic conjunctivitis and compared the expression level of Fc(epsilon)RI-beta with control conjunctivae using the anti-human Fc(epsilon)RI-beta antibody. METHODS: Fc(epsilon)RI-beta expression in giant papillae obtained from patients with atopic keratoconjunctivitis and vernal keratoconjunctivitis in control conjunctivae was evaluated by immunohistochemistry using anti-Fc(epsilon)RI-beta, -alpha, -gamma, and anti-human mast cell tryptase, anti-chymase, anti-basophil, and anti-CD1a antibodies. RESULTS: Statistical analyses revealed that the densities of Fc(epsilon)RI-beta(+) cells, Fc(epsilon)RI-alpha(+) cells, tryptase(+) cells, and Fc(epsilon)RI-beta(+)/tryptase(+) cells were significantly increased in giant papillae compared with controls. There were two types of Fc(epsilon)RI (alphabetagamma(2) and alphagamma(2)) on the mast cells of the giant papillae. The ratio of the Fc(epsilon)RI-beta(+) cell number/Fc(epsilon)RI alpha(+) cell number in the giant papillae (0.69 +/- 0.08 [mean +/- SD]) was significantly higher than that of the controls (0.07 +/- 0.16). Fc(epsilon)RI beta/tryptase double immunostaining revealed that 81% +/- 13% of tryptase(+) cells expressed Fc(epsilon)RI-beta. Fc(epsilon)RI-beta(+) cells were preferentially localized within and around epithelial tissue. The authors also found that Fc(epsilon)RI-beta was expressed by basophils but not by Fc(epsilon)RI alphagamma(2)-positive Langerhans cells in the giant papillae samples. CONCLUSIONS: Preferential Fc(epsilon)RI-beta expression observed in the mast cells and basophils of giant papillae suggests important roles of Fc(epsilon)RI beta in the pathophysiology of atopic keratoconjunctivitis and vernal keratoconjunctivitis. PMID- 19136705 TI - Visual morbidity due to inaccurate spectacles among school children in rural China: the See Well to Learn Well Project, report 1. AB - PURPOSE: Inadequately corrected refractive error is the leading cause of visual disability among children in China; inaccurate spectacles are a potential cause. The prevalence and visual impact of spectacle inaccuracy were studied among rural, secondary-school children, to determine the optimal timing for updating of refraction. METHODS: A random sample of children from years 1 and 2 in all junior and senior high schools in Fuyang Township, Guangdong Province, underwent ocular examination. All children who reported wearing glasses received cycloplegic refraction, vision assessment, and measurement of current spectacles. RESULTS: Among 3226 examined children, 733 (22.7%) reported owning spectacles. Refractive error and spectacle power were assessed for 588 (80.2%) children. They had a mean age of 15.0 +/- 1.6 years; 70.2% were girls, 83.3% had more than -1.5 D of myopia, and 17.9% had presenting vision < or = 6/12 in the better eye. The glasses of 48.8% of children were inaccurate by > or = 1 D; inaccuracy was > or = 2 D in 17.7%. Children with inaccurate glasses (> or = 1 D) had presenting vision in the better eye significantly (P < 0.001) worse than that of children with accurate glasses, and 30.3% had presenting acuity < or = 6/12. In multivariate models, younger age (P = 0.004), more myopic refractive error (P < 0.001), and having glasses > or = 1 year old (P = 0.04) were associated with inaccurate spectacles. DISCUSSION: Inaccurate spectacles are common and are associated with significant visual impairment among children in rural China. Reducing outdated glasses could lessen the visual burden, although refractive services may have to be offered on an annual basis for optimal benefit. PMID- 19136706 TI - Suppression of experimental autoimmune uveitis by angiotensin II type 1 receptor blocker telmisartan. AB - PURPOSE: Angiotensin II type 1 receptor (AT1-R) blockers are used widely for the treatment of patients with hypertension. Recent reports have suggested that AT1-R also plays a key role in various inflammatory conditions. The aim of this study was to examine whether blockade of AT1-R is effective in the suppression of murine experimental autoimmune uveoretinitis (EAU). METHODS: C57BL/6 mice were immunized with human interphotoreceptor retinoid binding protein-derived peptide 1-20 (hIRBP-p). Telmisartan, an AT1-R blocker, was administrated daily by intraperitoneal injection. On day 21 after immunization, the severity of EAU was assessed clinically and histopathologically. With the use of flow cytometry, the activation of draining lymph node (LN) cells was assessed by cell proliferation response against hIRBP-p and by the number of CD44(high) activated CD4(+) T cells present. In addition, mRNA expression of ICAM-1, MCP-1, and IFN-gamma in the eye was analyzed by reverse-transcriptase PCR, and the number of retinal adherent leukocytes was counted by retinal perfusion labeling. RESULTS: Telmisartan significantly suppressed EAU clinically and histopathologically. Intraocular mRNA expression of ICAM-1 and MCP-1 was downregulated, and the retinal adherent leukocyte counts were significantly decreased in telmisartan-treated mice compared with vehicle-treated mice. LN cell proliferative responses against hIRBP p and the number of CD44(high)CD4(+) T cells were remarkably reduced in telmisartan-treated mice. CONCLUSIONS: Systemic administration of telmisartan significantly suppressed EAU by the inhibition of antigen-specific T-cell activation in the LNs and of leukocyte adhesion in the retina. These results indicate that telmisartan may be a novel therapeutic regimen for patients with endogenous uveitis. PMID- 19136707 TI - Evaluation of ranibizumab-induced changes in high-resolution optical coherence tomographic retinal morphology and their impact on visual function. AB - PURPOSE: To evaluate the effects of intravitreal ranibizumab on retinal function and morphology and to identify a correlation between anatomy and function by using spectral domain optical coherence tomography (SDOCT). METHODS: Twenty-three patients affected by neovascular AMD received three injections of ranibizumab in three consecutive months and were monitored by assessment of best corrected visual acuity (BCVA), central retinal sensitivity (CRS) and morphologic changes at the level of the retina and the retinal pigment epithelium (RPE). The morphologic changes, identified by SDOCT segmentation, were mean retinal thickness (MRT), central retinal thickness (CRT), and the pathologic area (lesion area) of the RPE. RESULTS: BCVA increased from a mean 60.1 +/- 8.7 letters at baseline to 67.0 +/- 10.9 at month 3 (P = 0.0003). The CRS at the 0 degrees position increased from 2.8 +/- 3.1 dB at baseline to 4.0 +/- 5.7 at week 1, remaining stable until month 3. Absolute scotoma size decreased continuously from baseline to month 3, in a mean of 5.3 +/- 5.8 to 3.6 +/- 4.0 test point locations. By SDOCT, MRT decreased from 308.6 +/- 25.9 microm at baseline to 268.4 +/- 22.4 microm at month 3 (P = 0.0001). CRT was 365.8 +/- 84.9 and 254.9 +/- 95.1 microm at month 3 (P = 0.0002). The mean RPE lesion area was 6.0 +/- 3.0 mm(2) at baseline, which decreased to 5.0 +/- 3.1 mm(2) at month 3 (P = 0.115). The only significant correlation was identified between the lesion area and CRS. CONCLUSIONS: In ranibizumab therapy, the condition of the RPE lesion may be more relevant for visual function than the usual OCT parameters, retinal thickness. PMID- 19136708 TI - Depletion of passenger leukocytes from corneal grafts: an effective means of promoting transplant survival? AB - PURPOSE: To develop and compare effective strategies for depleting graft-derived passenger leukocytes that include antigen-presenting cells from corneal buttons and to assess the effectiveness of this strategy in promoting graft survival using a high-risk (HR) model of corneal transplantation. METHODS: Corneal buttons harvested from C57BL/6 mice were used in three ex vivo strategies of passenger leukocyte depletion. Two strategies involved storage in medium at different temperatures for prolonged periods. A third strategy used complement-dependent cytotoxicity (CDC) by treating the buttons with anti-CD45 mAb plus complement. Wholemount corneal buttons or cells from enzyme-digested corneas were analyzed using confocal microscopy or flow cytometry, respectively, for the pan-leukocyte surface marker CD45. HR host beds were created and used to evaluate the efficacy of passenger leukocyte depletion on transplant survival. RESULTS: Passenger leukocyte numbers in the buttons were significantly reduced by all three treatments. CDC was the most efficient strategy for passenger leukocyte depletion with 39% reduction (P < 0.00005) of CD45(+) cells, and negligible damage to the endothelial layer, achievable within 24 hours. However, passenger leukocyte depletion failed to improve HR graft longevity. CONCLUSIONS: Anti-CD45 antibody plus complement-mediated targeting of donor tissue is the most efficient way to deplete corneal passenger leukocytes and can considerably reduce the time required for cell depletion. However, depletion of graft passenger leukocytes does not have a significant effect on promoting graft survival even in the HR setting. PMID- 19136709 TI - A drug-eluting contact lens. AB - PURPOSE: To formulate and characterize a drug-eluting contact lens designed to provide extended, controlled release of a drug. METHODS: Prototype contact lenses were created by coating PLGA (poly[lactic-co-glycolic acid]) films containing test compounds with pHEMA (poly[hydroxyethyl methacrylate]) by ultraviolet light polymerization. The films, containing encapsulated fluorescein or ciprofloxacin, were characterized by scanning electron microscopy. Release studies were conducted in phosphate-buffered saline at 37 degrees C with continuous shaking. Ciprofloxacin eluted from the contact lens was studied in an antimicrobial assay to verify antimicrobial effectiveness. RESULTS: After a brief and minimal initial burst, the prototype contact lenses demonstrated controlled release of the molecules studied, with zero-order release kinetics under infinite sink conditions for over 4 weeks. The rate of drug release was controlled by changing either the ratio of drug to PLGA or the molecular mass of the PLGA used. Both the PLGA and the pHEMA affected release kinetics. Ciprofloxacin released from the contact lenses inhibited ciprofloxacin-sensitive Staphylococcus aureus at all time-points tested. CONCLUSIONS: A prototype contact lens for sustained drug release consisting of a thin drug-PLGA film coated with pHEMA could be used as a platform for ocular drug delivery with widespread therapeutic applications. PMID- 19136710 TI - Common and distinct pathways for cellular activities in FGF-2 signaling induced by IL-1beta in corneal endothelial cells. AB - PURPOSE: To determine the mechanism by which IL-1beta induces FGF-2 and to elucidate the signaling pathways of IL-1beta-induced FGF-2 in corneal endothelial cells (CECs). METHODS: Expression and/or activation of FGF-2, p38, ERK1/2, and Akt was analyzed by immunoblot analysis. Cell proliferation was measured by MTT assay. Pharmacologic inhibitors were used to block PI 3-kinase, p38, or ERK1/2. RESULTS: Brief stimulation of CECs with IL-1beta activated PI 3-kinase and p38 in a biphasic fashion. The first wave of activation, triggered by IL-1beta, involves the inductive activity of IL-1beta on FGF-2 production; the second wave of activation, triggered by the induced FGF-2, involves the promotion of cellular activities. In both pathways, p38 acts downstream to PI 3-kinase. The inductive activity of IL-1beta on FGF-2 is further evidenced by the conditioned medium, which contains a large amount of FGF-2. Stimulation of CECs with IL-1beta also activated ERK1/2 in a delayed fashion. The IL-1beta-induced FGF-2 exerted cellular activities using distinct pathways: the second wave of activation of PI 3-kinase and p38 was involved in cell migration, whereas cell proliferation was simultaneously stimulated by ERK1/2 and the second wave of PI 3-kinase. Likewise, the conditioned medium demonstrated cellular activities and pathways identical with those observed in cells treated with IL-1beta. CONCLUSIONS: These data suggest that CECs produce FGF-2 by IL-1beta stimulation through PI 3-kinase and p38. The IL-1beta-induced FGF-2 facilitates cell migration via PI 3-kinase and p38, whereas it stimulates cell proliferation using PI 3-kinase and ERK1/2 in parallel pathways. PMID- 19136711 TI - In situ fluorescence measurement of tear film [Na+], [K+], [Cl-], and pH in mice shows marked hypertonicity in aquaporin-5 deficiency. AB - PURPOSE: Tear film composition depends on water and ion transport across ocular surface epithelia and on fluid secretion by lacrimal glands. The purpose of this study was to establish in situ fluorescence methods to measure tear film ionic concentrations and pH in mice and to determine whether tear film composition is sensitive to deficiency of the major ocular surface aquaporin water channels. METHODS: Tear film ionic concentrations and pH were measured in anesthetized mice by ratio imaging fluorescence microscopy after topical application of ion/pH sensing, dual-wavelength fluorescent indicators. [Na(+)], [K(+)], and [Cl(-)] were measured with membrane-impermeant indicators developed by our laboratory, and pH was measured with bis(carboxyethyl)-carboxyfluorescein fluorescence conjugated dextran. Measurements were performed on wild-type mice and on knockout mice lacking aquaporins AQP1, AQP3, and AQP5. RESULTS: In wild-type mice, tear film [Na(+)] was 139 +/- 8 mM, [K(+)] was 48 +/- 1 mM, [Cl(-)] was 127 +/- 4 mM, and pH was 7.59 +/- 0.2 (SE; n = 5-8). pH did not differ significantly in the AQP knockout mice. [Na(+)] was increased by approximately twofold in AQP5 null mice (230 +/- 20 mM) and was greatly reduced after exposure of the ocular surface to a humidified atmosphere. [K(+)] was mildly reduced in AQP1 null mice. CONCLUSIONS: These results establish an in situ optical methodology to measure tear film [Na(+)], [K(+)], [Cl(-)], and pH in living mice, without the need for fluid sampling. Tear film hypertonicity in AQP5 deficiency is likely caused by reduced transcorneal water secretion in response to evaporative water loss. PMID- 19136712 TI - Waiting time for cataract surgery and its influence on patient attitudes. AB - PURPOSE: To characterize willingness to pay for private operations and preferred waiting time among patients awaiting cataract surgery in Hong Kong. METHODS: This was a cross-sectional survey. Subjects randomly selected from cataract surgical waiting lists in Hong Kong (n = 467) underwent a telephone interview based on a structured, validated questionnaire. Data were collected on private insurance coverage, preferred waiting time, amount willing to pay for surgery, and self reported visual function and health status. RESULTS: Among 300 subjects completing the interview, 144 (48.2%) were 76 years of age or older, 177 (59%) were women, and mean time waiting for surgery was 17 +/- 15 months. Among 220 subjects (73.3%) willing to pay anything for surgery, the mean amount was US$552 +/- 443. With adjustment for age, education, and monthly household income, subjects willing to pay anything were less willing to wait 12 months for surgery (OR = 4.34; P = 0.002), more likely to know someone having had cataract surgery (OR = 2.20; P = 0.03), and more likely to use their own savings to pay for the surgery (OR = 2.21; P = 0.04). Subjects considering private cataract surgery, knowing people who have had cataract surgery, using nongovernment sources to pay for surgery, and having lower visual function were willing to pay more. CONCLUSIONS: Many patients wait significant periods for cataract surgery in Hong Kong, and are willing to pay substantial amounts for private operations. These results may have implications for other countries with cataract waiting lists. PMID- 19136713 TI - Biphasic photoreceptor degeneration induced by light in a T17M rhodopsin mouse model of cone bystander damage. AB - PURPOSE: To evaluate light-induced retinal damage in transgenic T17M rhodopsin mice as a novel model for bystander cone damage during retinal degeneration. METHODS: Mouse eyes were exposed to bright white light (15,000 lux, 2.5 minutes). After exposure, electroretinography was performed on mice dark adapted for 12 or more hours at 0 to 5 days to test photoreceptor response or for 0 to 12 hours to test response recovery. Retinal cryosections were examined by TUNEL staining and outer nuclear layer thickness measurements. Cone morphology was assessed by peanut agglutinin staining in retinal flatmounts and cryosections. RESULTS: T17M retinal function and morphology changed rapidly after exposure to light. Scotopic and photopic electroretinogram responses declined progressively from 0.5 to 3 days. Scotopic response recovery peaked at 50% to 60% of the unilluminated response in 3 hours, indicating an early, rapid decline in scotopic signaling. Photopic responses were near normal or supernormal from 0 to 6 hours. Cell death peaked at 1 day, and outer nuclear layer thickness declined from 1 to 5 days. Disorganized cones were observed at 6 hours, intact and damaged cones were observed at 12 hours and 1 day, but only cone remnants were observed at 3 and 5 days. Light exposure had little to no effect on ERG responses in nontransgenic littermates and other retinal degeneration models. CONCLUSIONS: The time course of light-induced T17M retinal damage is biphasic, with an initial decline in rod function within hours followed by bystander cone and rod deterioration within days. The rapid and synchronous induction of damage in this model is attractive for characterizing bystander effects in retinal degeneration. PMID- 19136714 TI - Novel sutureless keratoplasty with a chemically defined bioadhesive. AB - PURPOSE: The purpose of this study was to evaluate sutureless keratoplasty using a chemically-defined bioadhesive (CDB) made from food or medical additives. METHODS: Sutureless automated lamellar therapeutic keratoplasty (ALTK) using a CDB was performed on three rabbit eyes. Allogenic lamellar graft was transplanted onto the recipient bed using either suture fixation or a sutureless technique using the CDB. Slit-lamp examination was performed at selected intervals to evaluate the grade of epithelialization and the corneal clarity. The rabbits were killed at 90 days after operation and the eyes processed for histology, electron microscopic examination, and immunohistochemistry for cytokeratins and cell junction-related proteins. RESULTS: Sutureless keratoplasty was successfully performed with appropriate handling time before the CDB gelatinized. All the glued grafts were rapidly epithelialized within 7 days, and thereafter remained clear and attached for 90 days. Histologic and ultrastructural findings on the sutureless group showed the normal feature of stromal and epithelial cells and the grafts to be closely adhered with no inflammatory or scarring changes on the interface. Immunohistochemistry of the epithelial cells on the sutureless group revealed a similar expression pattern to the control group. CONCLUSIONS: These results demonstrate that sutureless keratoplasty using the CDB is easy to perform, with reliable attachment and no fear of toxic effects or disease transmissions. The authors expect the CDB to become a major choice for corneal treatment, especially in lamellar keratoplasty, posterior keratoplasty, and amniotic membrane transplantation on corneas. PMID- 19136715 TI - Morphologic and histopathologic changes in the rabbit cornea produced by femtosecond laser-assisted multilayer intrastromal ablation. AB - PURPOSE: To observe morphologic and histopathologic changes in the midperiphery of the rabbit cornea produced by femtosecond laser-assisted multilayer intrastromal ablation, determine whether this method may be used to correct myopia, and study how the cornea heals when the epithelium is not injured. METHODS: The right eyes of 10 New Zealand White rabbits were used for the experiments. A 60-kHz femtosecond laser delivery system was used, and three lamellar layers of laser pulses were focused starting at a corneal depth of 180 microm and ending at 90 microm from the surface, with each successive layer placed 45 microm anterior to the previous layer. In the interface of the applanation contact lens cone, a 6-mm diameter aluminum circle was placed at the center to block the laser, and ablation was limited to the midperiphery of the cornea. The laser settings were spot/line separation, 10 microm; diameter, 8.5 mm; energy for ablating the stroma, 1.3 microJ. Topography examination was used to document changes in corneal power. Light microscopy, transmission electron microscopy (TEM), and confocal microscopy in vivo were applied to observe changes in the cornea. RESULTS: There was significant change in mean corneal power between baseline and postoperative month 3 (n = 8; P = 0.0001), with a decrease from 46.82 D to 44.42 D. There was no haze formation or refractive regression throughout the follow-up. There were no corneal structural abnormalities under light microscopy. Activated keratocytes and necrotic debris were visible under confocal microscopy. Fibroblasts were observed, and no myofibroblasts appeared under TEM. CONCLUSIONS: Multilayer intrastromal ablation by the femtosecond laser with intact epithelium in the midperiphery of the corneal stroma can flatten the cornea without causing haze formation or refractive regression. This procedure allows the cornea to heal differently than when traditional corneal refractive surgery is performed and the epithelium is damaged. PMID- 19136716 TI - Human corneal endothelial cell transplantation in a human ex vivo model. AB - PURPOSE: To determine the effects of incorporating superparamagnetic microspheres (SPMs) into cultured human corneal endothelial cells (HCECs) and to describe preliminary experiments of HCEC transplantation, facilitated by SPMs and an external magnetic field, in a human anterior segment ex vivo model. METHODS: HCECs were cultured as monolayers and incorporated with magnetite oxide SPMs (900, 300, and 100 nm) at different concentrations. Cell viability, migration toward a magnetic field, and light transmittance were measured after incorporation of the SPMs. HCEC transplantation into the eyes of human recipients was investigated by subjecting anterior segments in organ culture to an external magnetic field. Light and electron microscopy were used to assess HCEC attachment to corneal stroma. RESULTS: SPMs were incorporated into the cytoplasm of HCECs after overnight incubation. None of the SPMs affected the short-term viability of cultured HCECs (P > 0.14, n = 6) or their light transmittance (P > 0.06, n = 5), although there was a trend toward decreased transmittance with the higher concentration of 900-nm SPMs. Cell migration toward a magnetic field was higher for HCECs with incorporated SPMs than for HCECs without SPMs (P < or = 0.01, n = 6), with dose-response relationships evident for the 300- and 100-nm SPMs. SPMs facilitated the attachment of HCECs to the corneal stroma in the human anterior segment model with minimal change in intracameral (intraocular) pressure. CONCLUSIONS: SPMs facilitate migration of HCECs toward a magnetic source and attachment of cells to the corneal stroma without affecting cell viability or light transmittance. The human anterior segment model can be used to study HCEC transplantation. PMID- 19136717 TI - The puzzle of ligand binding to Corynebacterium ammoniagenes FAD synthetase. AB - In bacteria, riboflavin phosphorylation and subsequent conversion of FMN into FAD are carried out by FAD synthetase, a single bifunctional enzyme. Both reactions require ATP and Mg(2+). The N-terminal domain of FAD synthetase appears to be responsible for the adenylyltransferase activity, whereas the C-terminal domain would be in charge of the kinase activity. Binding to Corynebacterium ammoniagenes FAD synthetase of its products and substrates, as well as of several analogues, is analyzed. Binding parameters for adenine nucleotides to each one of the two adenine nucleotide sites are reported. In addition, it is demonstrated for the first time that the enzyme presents two independent flavin sites, each one related with one of the enzymatic activities. The binding parameters of flavins to these sites are also provided. The presence of Mg(2+) and of both adenine nucleotides and flavins cooperatively modulates the interaction parameters for the other ligands. Our data also suggest that during its double catalytic cycle FAD synthetase must suffer conformational changes induced by adenine nucleotide-Mg(2+) or flavin binding. They might include not only rearrangement of the different protein loops but also alternative conformations between domains. PMID- 19136718 TI - Lipin 2 is a liver-enriched phosphatidate phosphohydrolase enzyme that is dynamically regulated by fasting and obesity in mice. AB - Lipin 1 is a bifunctional intracellular protein that regulates fatty acid metabolism in the nucleus via interactions with DNA-bound transcription factors and at the endoplasmic reticulum as a phosphatidic acid phosphohydrolase enzyme (PAP-1) to catalyze the penultimate step in triglyceride synthesis. However, livers of 8-day-old mice lacking lipin 1 (fld mice) exhibited normal PAP-1 activity and a 20-fold increase in triglyceride levels. We sought to further analyze the hepatic lipid profile of these mice by electrospray ionization mass spectrometry. Surprisingly, hepatic content of phosphatidate, the substrate of PAP-1 enzymes, was markedly diminished in fld mice. Similarly, other phospholipids derived from phosphatidate, phosphatidylglycerol and cardiolipin, were also depleted. Another member of the lipin family (lipin 2) is enriched in liver, and hepatic lipin 2 protein content was markedly increased by lipin 1 deficiency, food deprivation, and obesity, often independent of changes in steady state mRNA levels. Importantly, RNAi against lipin 2 markedly reduced PAP-1 activity in hepatocytes from both wild type and fld mice and suppressed triglyceride synthesis under conditions of high fatty acid availability. Collectively, these data suggest that lipin 2 plays an important role as a hepatic PAP-1 enzyme. PMID- 19136719 TI - Diversity of serine hydrolase activities of unchallenged and botrytis-infected Arabidopsis thaliana. AB - Activity-based protein profiling is a powerful method to display enzyme activities in proteomes and provides crucial information on enzyme activity rather than protein or transcript abundance. We applied activity-based protein profiling using fluorophosphonate-based probes to display the activities of Ser hydrolases in the model plant Arabidopsis thaliana. Multidimensional protein identification technology and in-gel analysis of fluorophosphonate-labeled leaf extracts revealed over 50 Ser hydrolases, including dozens of proteases, esterases, and lipases, representing over 10 different enzyme families. Except for some well characterized Ser hydrolases like subtilases TPP2 and ARA12, prolyl oligopeptidase acylamino acid-releasing enzyme, serine carboxypeptidase-like SNG1 and BRS1, carboxylesterase-like CXE12, methylesterases MES2 and MES3, and S formylglutathione hydrolase, the majority of these serine hydrolases have not been described before. We studied transiently expressed SNG1 and investigated plants infected with the fungal pathogen Botrytis cinerea. Besides the down regulation of several Arabidopsis Ser hydrolase activities during Botrytis infection, we detected the activities of Botrytis-derived cutinases and lipases, which are thought to contribute to pathogenicity. PMID- 19136721 TI - Dr Miroslav Ryska: biographical notes. PMID- 19136720 TI - Functional unfolding of alpha1-antitrypsin probed by hydrogen-deuterium exchange coupled with mass spectrometry. AB - The native state of alpha(1)-antitrypsin (alpha(1)AT), a member of the serine protease inhibitor (serpin) family, is considered a kinetically trapped folding intermediate that converts to a more stable form upon complex formation with a target protease. Although previous structural and mutational studies of alpha(1)AT revealed the structural basis of the native strain and the kinetic trap, the mechanism of how the native molecule overcomes the kinetic barrier to reach the final stable conformation during complex formation remains unknown. We hypothesized that during complex formation, a substantial portion of the molecule undergoes unfolding, which we dubbed functional unfolding. Hydrogen-deuterium exchange coupled with ESI-MS was used to analyze this serpin in three forms: native, complexing, and complexed with bovine beta-trypsin. Comparing the deuterium content at the corresponding regions of these three samples, we probed the unfolding of alpha(1)AT during complex formation. A substantial portion of the alpha(1)AT molecule unfolded transiently during complex formation, including not only the regions expected from previous structural studies, such as the reactive site loop, helix F, and the following loop, but also regions not predicted previously, such as helix A, strand 6 of beta-sheet B, and the N terminus. Such unfolding of the native interactions may elevate the free energy level of the kinetically trapped native serpin sufficiently to cross the transition state during complex formation. In the current study, we provide evidence that protein unfolding has to accompany functional execution of the protein molecule. PMID- 19136722 TI - Collisions of slow ions C3Hn+ and C3Dn+ (n = 2-8) with room temperature carbon surfaces: mass spectra of product ions and the ion survival probability. AB - Collisions of C3Hn+ (n = 2-8) ions and some of their per- deuterated analogs with room temperature carbon (HOPG) surfaces (hydrocarbon-covered) were investigated over the incident energy range 13-45 eV in beam scattering experiments. The mass spectra of product ions were measured and main fragmentation paths of the incident projectile ions, energized in the surface collision, were determined. The extent of fragmentation increased with increasing incident energy. Mass spectra of even-electron ions C3H7+ and C3H5+ showed only fragmentations, mass spectra of radical cations C3H8*+ and C3H6*+ showed both simple fragmentations of the projectile ion and formation of products of its surface chemical reaction (H atom transfer between the projectile ion and hydrocarbons on the surface). No carbon-chain build-up reaction (formation of C4 hydrocarbons) was detected. The survival probability of the incident ions, S(a), was usually found to be about 1 2% for the radical cation projectile ions C3H8*+, C3H6*+, C3H4*+ and C3H2*+ and several percent up to about 20% for the even-electron projectile ions C3H7+, C3H5+, C3H3+. A plot of S(a) values of C1, C2, C3, some C7 hydrocarbon ions, Ar+ and CO2+ on hydrocarbon-covered carbon surfaces as a function of the ionization energies (IE) of the projectile species showed a drop from about 10% to about 1% and less at IE 8.5-9.5 eV and further decrease with increasing IE. A strong correlation was found between log S(a) and IE, a linear decrease over the entire range of IE investigated (7-16 eV), described by log S(a) = (3.9 +/- 0.5)-(0.39 +/- 0.04) IE. PMID- 19136723 TI - Deciphering Dorin M glycosylation by mass spectrometry. AB - The soft tick, Ornithodoros moubata, is a vector of several bacterial and viral pathogens including Borrelia duttoni, a causative agent of relapsing fever and African swine fever virus. Previously, a sialic acid-specific lectin Dorin M was isolated from its hemolymph. Here, we report on the complete characterization of the primary sequence of Dorin M. Using liquid chromatography coupled to mass spectrometry, we identified three different glycopeptides in the tryptic digest of Dorin M. The peptide, as well as the glycan part of all glycopeptides, were further fully sequenced by means of tandem mass spectrometry (MS2) and multiple stage mass spectrometry (MS3). Two classical N-glycosylation sites were modified by high-mannose-type glycans containing up to nine mannose residues. The third site bore a glycan with four to five mannose residues and a deoxyhexose (fucose) attached to the proximal N-acetylglycosamine. The microheterogeneity at each site was estimated based on chromatographic behavior of different glycoforms. The fourth, a non-classical N-glycosylation site (Asn-Asn-Cys), was not glycosylated, probably due to the involvement of the cysteine residue in a disulfide bridge. PMID- 19136724 TI - Intra-molecular cross-linking of acidic residues for protein structure studies. AB - Intra-molecular cross-linking has been suggested as a method of obtaining distance constraints that would help to develop structural models of proteins. Recent work published on intra-molecular cross-linking for protein structural studies has employed commercially available primary amine (lysine, the amino terminus) selective reagents. Previous work using these cross-linkers has shown that for several proteins of known structure, the number of cross-links that can be obtained experimentally may be small compared to what would be expected from the known structure, due to the relative reactivity, distribution and solvent accessibility of the lysines in the protein sequence. To overcome these limitations, we have investigated the use of cross-linking reagents that can react with other reactive side chains in proteins. We used 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide hydrochloride (EDC) to activate the carboxylic acid containing residues, aspartic acid (D), glutamic acid (E) and the carboxy terminus (O), for cross-linking reactions. Once activated, the DEO side chains can react to form "zero-length" cross-links with nearby primary amine containing residues, lysines (K) and the amino terminus (X), via the formation of a new amide bond. We also show that the EDC-activated DEO side chains can be cross linked to each other using dihydrazides, two hydrazide moieties connected by an alkyl cross-linker arm of variable length. Using these reagents, we have found three new "zero-length" cross-links in ubiquitin consistent with its known structure (M1-E16, M1-E18 and K63-E64). Using the dihydrazide cross-linkers, we have identified two new cross-links (D21-D32 and E24-D32) unambiguously. Using a library of dihydrazide cross-linkers with varying arm length, we have shown that there is a minimum arm length required for the DEO-DEO cross-links of 5.8 A. These results show that additional structural information can be obtained by exploiting new cross-linker chemistry, increasing the probability that the protein target will yield sufficient distance constraints to develop a structural model. PMID- 19136725 TI - Electronic properties of charge-tagged peptides upon electron capture. AB - We report a computational study of Ala-Lys (AK) and Lys-Ala (KA) dipeptide ions furnished with fixed-charged pyridinium groups that were attached by amide linkers to the N-terminal amino groups. Cation-radicals from one-electron reduction of the doubly charged AK and KA peptide conjugates showed various extents of unpaired electron density being delocalized between the pyridine and peptide moieties. The delocalization depended on the local recombination energies (RE(loc)) of the charged groups. The RE(loc) of the pyridine moieties were modified by introducing electron-donating substituents (CH(3), OCH(3), and N(CH(3))(2)). The RE(loc) of the peptide moieties were found to depend on the peptide conformation and internal solvation of the Lys ammonium groups. Substantial electron delocalization was found for combinations of pyridine substituents and peptide conformers with closely matched RE(loc), such as 4 dimethylamino-pyridine and internally solvated Lys ammonium or unsubstituted pyridine and free (unsolvated) Lys ammonium. The dissociation (DeltaH(diss)) and transition state energies (E(TS)) for the loss of the pyridine ring from the conjugates were found to be DeltaH(diss) = 34-36 kJ mol(-1) and E(TS) = 67-69 kJ mol(-1) for the unsubstituted pyridine moieties, but did not depend much on the peptide sequence. PMID- 19136726 TI - Aberrant processing forms of lung surfactant proteins SP-B and SP-C revealed by high-resolution mass spectrometry. AB - The mutation (g.1286T>C) of the pulmonary surfactant-associated protein C gene (SFTPC) leads to the I73T substitution in the precursor protein (pro-SP-C) and results in interstitial lung disease with the histological pattern of non specific interstitial pneumonia and pulmonary alveolar proteinosis. Central for the disease is the abnormal processing of the SP-C pro-protein to mature SP-C; however little is known about the nature of intermediates and processing products. We report here the application of high resolution Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry to the characterization of processing intermediates of hydrophobic pulmonary surfactant proteins SP-B and SP C in intra- alveolar surfactant material of a patient with I73T mutation. SP-C and SP-B processing forms were separated from broncho-alveolar lavage fluid using chloroform/methanol extraction and sodium dodecyl sulfate poly acrylamide gel electrophoreis, detected by Western blot and identified by electrospray- and matrix-assisted laser desorption/ionization-FT-ICR mass spectrometry. The mass spectrometric and immuno-analytical results show the intra-alveolar accumulation of an aberrant C-terminal SP-C processing products in which the mature SP-C protein part is missing and aberrant processing intermediates of SP-B. PMID- 19136727 TI - Application of silicon nanowires and indium tin oxide surfaces in desorption electrospray ionization. AB - Two nanostructured surfaces are introduced as advantageous substrates for desorption electrospray ionization mass spectrometry (DESI-MS). Nano-assisted laser desorption/ionization (NALDI) plates coated with silicon nanowires (SiNWs) and indium tin oxide (ITO) layers on glass are both conductive non-polar surfaces that were originally designed as superior substrates for matrix-free laser desorption/ionization. In this study, NALDI/SiNWs and ITO were tested as potentially useful DESI substrates for selected model analytes (cyclosporine, beauverolide, surfactin and nystatin). Both nanostructured surfaces produced more intense and longer-lasting signals than other tested surfaces (polytetrafluoroethylene, glass, polymethylmethacrylate and chromatography paper). PMID- 19136728 TI - Negative ion ultraviolet matrix-assisted laser desorption ionization mass spectrometry and post source decay of glycosyl esters of nucleoside pyrophosphates. AB - Six different glycosyl esters of nucleoside pyrophosphates (monosaccharide nucleotides) were analyzed by means of matrix-assisted laser desorption/ionisation reflectron time-of-flight mass spectrometry (MALDI/RToF MS) in the negative ion mode. Several matrices were evaluated and 3-hydroxypicolinic acid as well as alpha-cyano-4- hydroxycinnamic acid (CHCA) turned out to be the matrices of choice applying the thin layer technique to obtain maximum sensitivity for deprotonated molecular ion detection and maximal fragmentation particular with CHCA. The determination of the molecular mass with a mass accuracy below 0.1% was feasible with sample amounts in the lower femtomole range applying a MALDI desk-top mass spectrometer. A further important refinement of this technique was the use of post source decay (PSD) fragment ion analysis with a curved field reflector (which means no stepping of the reflector voltage). Detailed structural information of the six selected monosaccharide nucleotides could be obtained with PSD and differences in the fragmentation pattern were used to distinguish them. This method (based on molecular mass and PSD fragment ion analysis) has been applied to verify the presence of a glycosyl ester of nucleoside pyrophosphate in samples from Saccharomyces cerevisiae. PMID- 19136729 TI - Nano-desorption electrospray and kinetic method in chiral analysis of drugs in whole human blood samples. AB - A home-made nano-desorption electrospray ionization (nano-DESI) device and the kinetic method were tested in chiral analysis of model clinical samples containing enantiomers of one of three pharmaceutically important compounds: dihydroxyphenylalanine (DOPA), ephedrine and ibuprofen. The initial evaluation of chiral systems was carried out by direct infusion of solution mixtures (analyte/central metal/chiral reference ligand) to a standard electrospray ionization (ESI) source. Cu(II) was used as a central metal for all analytes, L phenylalanine was applied as a chiral reference ligand for DOPA, whereas L tryptophan was used for the other two analytes. Then, the ESI source was substituted by a nano-DESI source and dried spots of 1 microL samples of whole human blood spiked with individual drugs were successfully analyzed without any pre-treatment. Irrespective of a laborious initial nano-DESI set-up, the combination of the kinetic method with nano-desorption electrospray has, for the first time, been demonstrated as a promising tool for chiral analysis of drugs in blood samples. PMID- 19136730 TI - Analysis of complex oligosaccharides using graphitized carbon liquid chromatography/mass spectrometry. AB - The sugar fraction of alpha-1 acid glycoprotein (AGP) was studied using porous graphitized carbon (PGC) chromatography coupled to mass spectrometry. Electrospray ionization provides efficient control over fragmentation; at low collision energy only molecular species were observed, allowing accurate oligosaccharide profiling. PGC chromatography was useful separating 18 sugars differing in monosaccharide composition. Most of these were separated into several isomeric forms; altogether 49 different oligosaccharides were found in AGP. PMID- 19136731 TI - Comparing the habituation of late auditory evoked potentials to loud and soft sound. AB - The objective fitting of hearing aids and cochlear implants remains a challenge. In particular, the determination of whether sound is perceived as too loud or comfortable represents an unsolved problem in noncooperative patients. In a first step of an ongoing study, we assess the feasibility of habituation correlates in late auditory evoked potentials (LAEPs) to discriminate between a soft sound (SS) of 50 dB SPL and a loud sound (LS) of 100 dB SPL. We applied a new sweep-to-sweep time-scale coherence measure to analyse the habituation in LAEPs, i.e., relative changes within sweep sequences. From the comparison between both stimulation levels, a total discrimination of responses to SS and LS in the individual normal hearing subject was possible. As just relative changes in SS and LS sweep sequences were considered, purely exogenously driven morphological alternations in the responses such as intensity related amplitude and latency changes were excluded from the analysis. It is concluded that the proposed method allows for the reliable detection of auditory habituation and differentiation of SS from LS. The proposed scheme might provide an electrophysiological measurement and signal processing framework for the objective detection of the most comfortable loudness level and can be used in further, more clinically oriented studies. PMID- 19136732 TI - Impedance-based tissue discrimination for needle guidance. AB - Measurement of electrical impedance can discriminate between tissues of different electrical properties. A measurement system with adequate spatial resolution focused on a volume around the tip of a needle or other invasive clinical equipment can be used to determine in which type of tissue the tip is positioned. We have measured the sensitivity zone of a needle electrode with an active electrode area of 0.3 mm(2), and measured impedance spectra in porcine tissue in vivo. Small electrode impedance data will be influenced by electrode polarization impedance (EPI) at low frequencies. To refine existing methods for needle guidance with higher spatial resolution, we have used multivariate analysis and new interpretations of EPI, and tissue data gathered with selected needle electrodes. The focus of this study is on discrimination between muscle and fat/subdermis for drug administration, but our results also indicate that these refinements will facilitate new clinical applications for impedance-based needle guidance in general. PMID- 19136733 TI - Closed-form solutions for the optimum equivalence of first-order compartmental models and their implications for classical models of closed-circuit anesthesia. AB - Given a function that describes the uptake of a substance into the body with time, an analytical technique is described which transforms that function into a model of parallel first-order compartments that converges to the same uptake profile as the number of compartments is increased. The fitting of the compartmental model to the given uptake function is optimized to minimize the squared error. A necessary condition of the analytical method is that the uptake function be capable of being successively integrated at least as many times as the number of desired compartments. The uptake function should also be monotonically decreasing as all parallel first-order compartment models predict monotonically decreasing uptake. We applied this technique and ascertained the compartmental structure of the Severinghaus relationship, a longstanding observation in the field of clinical anesthesia that the uptake of nitrous oxide follows an inverse-square-root of time profile. The Severinghaus relationship is numerically poorly behaved at a time of zero elapsed minutes, predicting an instantaneously infinite uptake. Nevertheless, modeling of the first minute of anesthesia is necessary for characterizing the initial induction of anesthesia and methods of maintaining closed-circuit anesthesia such as the unit dose method. Using solely analytical methods, solutions for the compartmental properties of a mammillary model that matches the Severinghaus relationship for any expressed time interval are produced. These properties are compared to currently accepted values for the uptake of nitrous oxide. When matched to the Severinghaus relationship in the range of 0-100 min with a three-compartment model, we identified time constants of 0.28, 4.69 and 33.49 min with associated apparent volumes of 1.44, 2.14 and 7.97 l, respectively. The time constants in particular contrast to our earlier findings for the range of 1-100 min (1.46, 7.41 and 42.0 min). Our earlier findings were well matched to published time constants for tissues in classical pharmacokinetic models for volatile uptake. Consequently, we conclude that rigid adherence to the Severinghaus relationship from a time of zero minutes may lead to the over-administration of anesthetic agent due to an implicit mischaracterization of the relevant compartmental properties. PMID- 19136735 TI - The consistency of ordinary least-squares and generalized least-squares polynomial regression on characterizing the mechanomyographic amplitude versus torque relationship. AB - The primary purpose of this study was to examine the consistency of ordinary least-squares (OLS) and generalized least-squares (GLS) polynomial regression analyses utilizing linear, quadratic and cubic models on either five or ten data points that characterize the mechanomyographic amplitude (MMG(RMS)) versus isometric torque relationship. The secondary purpose was to examine the consistency of OLS and GLS polynomial regression utilizing only linear and quadratic models (excluding cubic responses) on either ten or five data points. Eighteen participants (mean +/- SD age = 24 +/- 4 yr) completed ten randomly ordered isometric step muscle actions from 5% to 95% of the maximal voluntary contraction (MVC) of the right leg extensors during three separate trials. MMG(RMS) was recorded from the vastus lateralis during the MVCs and each submaximal muscle action. MMG(RMS) versus torque relationships were analyzed on a subject-by-subject basis using OLS and GLS polynomial regression. When using ten data points, only 33% and 27% of the subjects were fitted with the same model (utilizing linear, quadratic and cubic models) across all three trials for OLS and GLS, respectively. After eliminating the cubic model, there was an increase to 55% of the subjects being fitted with the same model across all trials for both OLS and GLS regression. Using only five data points (instead of ten data points), 55% of the subjects were fitted with the same model across all trials for OLS and GLS regression. Overall, OLS and GLS polynomial regression models were only able to consistently describe the torque-related patterns of response for MMG(RMS) in 27-55% of the subjects across three trials. Future studies should examine alternative methods for improving the consistency and reliability of the patterns of response for the MMG(RMS) versus isometric torque relationship. PMID- 19136734 TI - The use of microelectrode array (MEA) to study the protective effects of potassium channel openers on metabolically compromised HL-1 cardiomyocytes. AB - The microelectrode array (MEA) was used to evaluate the cardioprotective effects of adenosine triphosphate sensitive potassium (K(ATP)) channel activation using potassium channel openers (KCOs) on HL-1 cardiomyocytes subjected to acute chemically induced metabolic inhibition. Beat frequency and extracellular action potential (exAP) amplitude were measured in the presence of metabolic inhibitors (sodium azide (NaN(3)) or 2-deoxyglucose (2-DG)) or KCOs (pinacidil (PIN, a cyanoguanidine derivative, activates sarcolemmal K(ATP) channels) or SDZ PCO400 (SDZ, a benzopyran derivative, activates mitochondrial K(ATP) channels)). The protective effects of these KCOs on metabolically inhibited HL-1 cells were subsequently investigated. Signal shapes indicated that NaN(3) and 2-DG reduced the rate of the sodium (Na(+)) influx signal as reflected by a reduction in beat frequency. PIN and SDZ appeared to reduce both rate of depolarization and extent of the Na(+) influx signals. Pre-treating cardiomyocytes with PIN (0.1 mM), but not SDZ, prevented the reduction of beat frequency associated with NaN(3)- or 2 DG-induced metabolic inhibition. The exAP amplitude was not affected by either KCO. The cardioprotective effect of PIN relative to SDZ may be due to the opening of different K(ATP) channels. This metabolic inhibition model on the MEA may provide a stable platform for the study of cardiac pathophysiology in the future. PMID- 19136736 TI - Pathology and diagnostic options of lower limb compartment syndrome. AB - BACKGROUND: The indication of surgical treatment in lower limb compartment syndrome mostly depends on the clinical signs which can be often uncertain, resulting in delayed insufficient intervention. AIM: The aim of the study was to evaluate the progression of compartment syndrome by measuring of intracompartmental pressure and monitoring of decreased tissular oxygenation, indicating an insufficient secondary microcirculation. MATERIALS AND METHODS: 16 patients were examined in our study (12 males, 4 females, mean age: 62.7+/-9.5 years), who underwent acute lower limb revascularization surgery for a critical (lasting more than 4 hours) limb ischemia. The indications were: 5 iliac artery embolizations and 11 femoral artery occlusions. After revascularization, on the second postoperative day, we detected significant lower limb edema and swelling of several grade. To monitor the elevated intracompartmental pressure (ICP) and to evaluate the extremital circulation, we used KODIAG pressure meter and the tissular oxygen saturation (StO2) was measured by near-infrared-spectroscopy. RESULTS: In 12 cases the ICP exceeded the critical 40 mmHg. In these patients the average StO2 was 50-53%, in spite of complete recanalization. In these cases we made urgent, semi-open fasciotomy. In 4 cases, where the clinical aspect showed compartment syndrome, the measured parameters did not indicate a surgical intervention (ICP: 25-35 mmHg, StO2: around normal). SUMMARY: A novel approach in our examination is that, besides empirical therapeutic guidelines generally applied in clinical practice, we established an objective, parameter-based ("evidence based medicine") surgical indication strategy for the lower limb compartment syndrome. Our parameter results produced by the above pressure and saturation measurements help the clinicians to decide between conservative and operative treatment of the disease. PMID- 19136738 TI - Effects of a herbal medicine, Hippophae rhamnoides, on cardiovascular functions and coronary microvessels in the spontaneously hypertensive stroke-prone rat. AB - The dry fruits of Hippophae rhamnoides (so-called "Saji" or "Sea buckthorn") are used in China as a herbal medicine. The present work studied the effects on microvessels in the left ventricular wall, hematological parameters, cardiovascular performance and plasma constituents in spontaneously hypertensive stroke-prone rats (SHRSP/EZO) treated with Hippophae for 60 days. Analyses showed that the powder made of dry Hippophae fruits contains the vitamins C, B1, B2 and E, provitamin A, rutin, serotonin, cytosterol, selenium and zinc, among other constituents. The experimental rats were fed ad libitum with blocks of rat chow supplemented with Hippophae powder at a concentration of 0.7 g/kg in rat powder chow, while control rats were unsupplemented chow. The mean arterial blood pressure, heart rate, total plasma cholesterol, triglycerides, and glycated hemoglobin were significantly decreased by the Hippophae treatment. The arteriolar capillary portions of microvessels expressing alkaline phosphatase decreased, but there was a trend for an increase in the total capillary density. It was concluded that Hippophae fruits improved the metabolic processes accompanied by reduction of hypertensive stress on the ventricular microvessels. PMID- 19136737 TI - Sampling time after tourniquet removal affects erythrocyte deformability and aggregation measurements. AB - Venipuncture procedures are widely thought to influence biochemical, hematological or hemorheological measurements. In line with the preparation of the new Guidelines for the standardization of hemorheological measurement, we compared various blood rheological parameters (i.e., red blood cell deformability and aggregation indices) assessed in blood samples obtained after 5, 30, 60 and 90 s following the tourniquet removal and a blood sample obtained without applying a tourniquet (control sample). A slight but significant improvement in red blood cell (RBC) deformability after the removal of tourniquet compared to the control sample was observed. RBC deformability was maximal in the samples obtained 30 s after tourniquet removal and remained slightly higher than the control in the following samples (at 60 and 90 s after tourniquet removal). The aggregation index (AI) decreased with time after tourniquet removal reaching significantly lower values than the control at 90 s after tourniquet removal. This finding was supported by a greater half time for RBC aggregation in the samples obtained 60 and 90 s after tourniquet removal. In conclusion, this study revealed that RBC deformability and aggregation might be significantly altered in the samples obtained after the application and removal of a tourniquet, as a part of the blood sampling procedure. Recommendation "remove the tourniquet at least 5 s prior to the start of blood sampling" may need to be revised. PMID- 19136739 TI - Comparative analysis of chosen hemorheological methods in a group of stroke patients. AB - Blood viscosity is one of the most important factors determining the blood flow. In this report we describe changes in whole blood viscosity in a group of patients after stroke measured as a function of the shear rate (flow curve) and also blood visco-elastic properties measured using oscillatory methods (dynamical mechanical analysis). The principle of the latter method is based on the measurement of the amplitude and phase of the sample oscillations driven by controlled harmonic strain. All measurements were performed by means of a rotary oscillatory rheometer Contraves LS-40. PMID- 19136740 TI - Influence of risk factors on nitric oxide metabolites at the initial stage of juvenile acute myocardial infarction. PMID- 19136741 TI - Hemorheological changes in solid tumor patients after treatment with recombinant erythropoetin. AB - The subject of this study was the effect of erythropoetin (Epoetin) treatment of anemic patients (n=30) with solid tumors on parameters of hemorheological profile. Both prior to and following Epoetin treatment (10,000 units subcutaneously thrice weekly) for four weeks hemorheological measurements included plasma and red blood cell (RBC) suspension viscosity; high and low shear whole blood viscosity; hematocrit (Hct), hemoglobin, RBC aggregation (RBCA) and deformation. It was found that the patients had reduced Hb up to 92.96+/-2.99 g/l, and the Hct - 28.2%. These parameters were significant increased after four weeks of Epoetin treatment: Hb by 28% (p<0.01) and Hct by 31% (p<0.01). In macrorheological part of the profile, both the high shear blood viscosity, and the low one were increased by 23 and 27% (p<0.05), respectively. These changes were mainly associated with the Hct rise. As for microrheological part of profile after Epoetin treatment, RBCA was decreased by 25% (p<0.05). While the red cell rigidity index (Tk) was lowered only slightly (by 8%). RBC incubation with Epoetin (10.0 I.E./ml) was accompanied by 10% decrease of Tk. It is important to note that before Epoetin treatment incubation with it led to decrease of RBCA by 30% (p<0.05), whereas after four week of the treatment period a 27% (p<0.05) rise of aggregation was found in majority of patients. Thus these results suggest that Epoetin is an effective, safe and convenient therapy for the management of anaemia in patients with cancer. And this drug has a moderate positive hemorheological effect on RBC and on the microrheological properties in particular. These data suggested that Epoetin has an effect on the membrane properties regulating RBC aggregation and deformation and affects by the signal transduction system, including Ca2+-dependant signaling pathway and tyrosine kinase and phosphotase activity change. PMID- 19136742 TI - Erythrocyte aggregation in rheumatoid arthritis: Cell and plasma factor's role. AB - Increase in erythrocyte aggregation (EA) is pathognomonic for rheumatoid arthritis (RA), and its estimation through erythrocyte sedimentation rate (ESR) is part of DAS 28-4 activity diagnosis, with low correlation with EA and that does not discriminate the contribution of cell factors that increase aggregation. OBJECTIVE: To analyse cell and plasma factors that might be involved in EA increase, to understand how RA affects blood components, thus modifying blood fluid behavior. METHODOLOGY: One hundred women presenting active RA were compared with age-matched controls (C). EA was measured by transmitted light, obtaining two parameters: 2k2n0, characterizing the aggregation process kinetics and s0/n0, estimating aggregates size. Cell factors assays: erythrocyte deformability, by filtration through nucleopore membranes, cell shape, by microscopy, and membrane fluidity by EPR. Plasma: total proteins and CRP, albumin, fibrinogen (Fb), by gravimetry, and IgG and IgM by single radial immuno-diffusion. RESULTS: AR and C (x+/-SE). 2k2n0: 31.83+/-2.84, 23.75+/-1.91; s0/n0: 0.92+/-0.05, 0.87+/-0.04. Rigidity index (RI): 14.79+/-4.71, 6.92+/-1.31. Morphological index: 0.28+/-0.03, 0.30+/-0.05, n.s. Fb (mg/dl): 382+/-80, 299+/-70. IgG (mg/dl): 1580+/-219, 1296+/ 158; IgM (mg/dl) 233+/-28, 183+/-23; albumin (g/dl) 3.84+/-0.44, 3.77+/-0.51 n.s. p<0.05 accepted. Correlations: 2k2n0 vs. Fb r=0.66; s0/n0 vs. Fb r=0.51; 2k2n0 vs. Igs r=0.65; s0/n0 vs. Igs r=0.56. 2k2n0 vs. RI r=-0.59; s0/n0 vs. RI=-0.52, p<0.05. CONCLUSIONS: Plasma factors, Igs and Fb increased aggregation, since RI is altered, this reduces the process efficiency regarding aggregation. Patients with active RA present an increased EA, with values modifications associated with the activity index DAS 28-4, thus becoming an RA activity indicator. PMID- 19136743 TI - Cutaneous vasomotion in patients with chronic venous insufficiency and the influence of compression therapy. AB - We investigated in patients with chronic venous insufficiency (CVI) and after compression therapy the fluxmotion within characteristic frequency bands, which were described earlier by Bracic and Stefanovska (Bull. Math. Biol. 60 (1998), 919-935).Therefore, the frequency spectra of laser Doppler flux data of the 36 patient's legs were compared with 41 legs of healthy subjects. In addition, 14 patients with CVI wore a compression stocking (interface pressure: 25-32 mmHg) or compression bandages and were measured after 4 weeks therapy. Data were analyzed by means of a Wavelet packet transformation (a combination of the Daubechies filter of order 4 and the Haar filter).We found significant differences between the patients and the healthy subjects in the frequency intervals of myogenic 0.06 0.16 Hz, respiratory 0.16-0.6 Hz and heart activity 0.6-1.6 Hz (p<0.05, Mann Whitney U test). Furthermore, the main energy peak height in these frequency intervals increased with the severity of venous disease and was highest in patients with venous leg ulceration. Compression therapy had a significant influence in myogenic vessel activity, which has been proved by a positive frequency shift of 20% (p=0.007, one-sided by the exact Wilcoxon test).In venous disease fluxmotion was increased. Compression therapy over a period of 4 weeks improved myogenic vessel activity. PMID- 19136744 TI - Erythrocyte aggregability and AB0 blood groups. AB - It is not established whether there is an association between erythrocyte aggregation and AB0 blood type, as glycophorins carry sialic acid which is responsible for the negative erythrocyte surface charge and the antigenicity for AB0 blood groups. We have determined erythrocyte aggregation by means of the Myrenne aggregometer in 114 healthy volunteers, along with plasma lipids, fibrinogen and AB0 blood groups. No differences in erythrocyte aggregation (EA0 and EA1) were observed when subjects with 0 (n = 45) and non-0 (n=69) blood group were compared (P = 0.624 and P = 0.838, respectively). Fibrinogen was statistically lower in 0 group compared with non-0 group (P = 0.012). Erythrocyte aggregation (EA0 and EA1) correlated significantly with both lipids and fibrinogen (P < 0.01). When erythrocyte aggregation was dichotomized as EA1 > or = 8, no association was found with 0 and non-0 blood groups (P > 0.05) but it was associated with high lipid levels: T-Chol > 220 mg/dl, TG > 175 mg/dl and high fibrinogen levels > 300 mg/dl (P = 0.035; P = 0.030; P = 0.010, respectively). Erythrocyte aggregation does not seem to be associated with blood groups, but rather with plasma lipids and fibrinogen. PMID- 19136745 TI - Influence of controlled vascular training on the pain free walking distance and plasmaviscosity in patients suffering from peripheral arterial occlusive disease. AB - Does controlled vascular training influence plasmaviscosity and the pain free walking distance in patients with peripheral arterial occlusive disease (paod) Fontaine stage II?37 patients, 24 men and 13 women with a mean age of 64.5 years SD 8.5 took part in ambulant vascular training over a period of 12 months.Before, after 6 months and after 12 months, pain free (pfwd) and maximum walking distance (mwd) was measured using a standardized treadmill program. Also ankle-brachials systolic pressure index (a-bspi), transcutaneous oxygen tension (tcPO2) and plasmaviscosity (pv) were measured.Pfwd increased from 212 SD 143 m to 371 SD 249 m (p<0.02). TcPO2 increased during training, but without statistically significance. A-bspi increased between the first 6 months of training statistically significant. Before training pv was 1.31 mPa s SD 0.10, after training period of 6 months it was 1.27 mPa s SD 0.11 (p=0.06) and 12 months later it was 1.28 SD 0.11 mPa s (p=0.35). The improvement of pfwd and the decrease of pv correlates (r=-0.39, p=0.05).In most patients, arterial vascular training improves pfwd and mwd. Simultaneously to the increase of the walking distances plasmaviscosity decreases and crurobrachial indexes increases. We found a coupling between improvement of pfwd and pv. Pv seems to participate in improvement of leg hemodynamics in patients with paod. PMID- 19136746 TI - Hemorheological abnormalities in stable angina and acute coronary syndromes. PMID- 19136747 TI - Thoracic epidural anaesthesia in cardiac surgery--the current standing. PMID- 19136749 TI - Effect of single intraoperative dose of amiodarone in patients with rheumatic valvular heart disease and atrial fibrillation undergoing valve replacement surgery. AB - Maintenance of sinus rhythm (SR) is superior to rate control in atrial fibrillation (AF). In order to achieve SR, we administered single-dose intravenous amiodarone intraoperatively and evaluated its effect on conversion of rheumatic AF to SR in patients undergoing valvular heart surgery. Patients were randomly assigned to amiodarone (n = 42) or control (n = 40) group in a double blind manner. The amiodarone group received amiodarone (3 mg/kg) intravenously prior to the institution of cardiopulmonary bypass and the control group received the same volume of normal saline. In the amiodarone group, the initial rhythm after the release of aortic cross clamp was noted to be AF in 14.3% (n = 6) and remained so in 9.5% (n = 4) of patients till the end of surgery. In the control group, the rhythm soon after the release of aortic cross clamp was AF in 37.5% (n = 15) (p = 0.035) and remained so in 32.5% (n = 13) of patients till the end of surgery (p = 0.01). At the end of first post-operative day 21.4% (n = 9) of patients in amiodarone group and 55% (n = 22) of patients in control group were in AF (p = 0.002). The requirement of cardioversion/defibrillation was 1.5 (+/ 0.54) in amiodarone group and 2.26 (+/-0.73) in the control group (p = 0.014), and the energy needed was 22.5 (+/-8.86) joules in the amiodarone group and 40.53 (+/-16.5) in the control group (p = 0.008). A single intraoperative dose of intravenous amiodarone increased the conversion rate of AF to normal sinus rhythm, reduced the need and energy required for cardioversion/defibrillation and reduced the recurrence of AF within one day. PMID- 19136748 TI - Halogenated anaesthetics and cardiac protection in cardiac and non-cardiac anaesthesia. AB - Volatile anaesthetic agents have direct protective properties against ischemic myocardial damage. The implementation of these properties during clinical anaesthesia can provide an additional tool in the treatment or prevention, or both, of ischemic cardiac dysfunction in the perioperative period. A recent meta analysis showed that desflurane and sevoflurane reduce postoperative mortality and incidence of myocardial infarction following cardiac surgery, with significant advantages in terms of postoperative cardiac troponin release, need for inotrope support, time on mechanical ventilation, intensive care unit and overall hospital stay. Multicentre, randomised clinical trials had previously demonstrated that the use of desflurane can reduce the postoperative release of cardiac troponin I, the need for inotropic support, and the number of patients requiring prolonged hospitalisation following coronary artery bypass graft surgery either with and without cardiopulmonary bypass. The American College of Cardiology/American Heart Association Guidelines recommend volatile anaesthetic agents during non-cardiac surgery for the maintenance of general anaesthesia in patients at risk for myocardial infarction. Nonetheless, evidence in non-coronary surgical settings is contradictory and will be reviewed in this paper together with the mechanisms of cardiac protection by volatile agents. PMID- 19136750 TI - Changes in near infrared spectroscopy during deep hypothermic circulatory arrest. AB - Monitoring cerebral oxygenation with near infrared spectroscopy may identify periods of cerebral desaturation and thereby the patients at risk for perioperative neurocognitive issues. Data regarding the performance of near infrared spectroscopy monitoring during deep hypothermic circulatory arrest are limited. The current study presents data regarding use of a commercially available near infrared spectroscopy monitor during deep hypothermic circulatory arrest in paediatric patients undergoing surgery for congenital heart disease. The cohort included 8 patients, 2 weeks to 6 months of age, who required deep hypothermic circulatory arrest for repair of congenital heart disease. The baseline cerebral oxygenation was 63 +/- 11% and increased to 88 +/- 7% after 15 min of cooling to a nasopharyngeal temperature of 17-18 degrees C on cardiopulmonary bypass. In 5 of 8 patients, the cerebral oxygenation value had achieved its peak value (either >or=90% or no change during the last 2-3 min of cooling on cardiopulmonary bypass). In the remaining 3 patients, additional time on cardiopulmonary bypass was required to achieve a maximum cerebral oxygenation value. The duration of deep hypothermic circulatory arrest varied from 36 to 61 min (43.4 +/- 8 min). After the onset of deep hypothermic circulatory arrest, there was an incremental decrease in cerebral oxygenation to a low value of 53 +/ 11%. The greatest decrease occurred during the initial 5 min of deep hypothermic circulatory arrest (9 +/- 3%). Over the entire period of deep hypothermic circulatory arrest, there was an average decrease in the cerebral oxygenation value of 0.9% per min (range of 0.5 to 1.6% decline per minute). During cardiopulmonary bypass, cooling and deep hypothermic circulatory arrest, near infrared spectroscopy monitoring followed the clinically expected parameters. Such monitoring may be useful to identify patients who have not achieved the highest possible cerebral oxygenation value despite 15 min of cooling on cardiopulmonary bypass. Future studies are needed to define the cerebral oxygenation value at which neurological damage occurs and if interventions to correct the decreased cerebral oxygenation will improve perioperative outcomes. PMID- 19136751 TI - Role of cardiac biomarkers (troponin I and CK-MB) as predictors of quality of life and long-term outcome after cardiac surgery. AB - Perioperative and postoperative morbidity and mortality associated with cardiac surgery affect both the outcome and quality of life. Markers such as troponin effectively predict short-term outcome. In a prospective cohort study in a University Hospital we assessed the role of cardiac biomarkers, also as predictors of long-term outcome and life quality after cardiac surgery with a three-year follow-up after conventional heart surgery. Patients were interviewed via phone calls with a structured questionnaire examining general health, functional status, activities of daily living, perception of life quality and need for hospital readmission. Descriptive statistics and multivariate analysis were performed. Out of 252 consecutive patients, 8 (3.2%) died at the three years follow up: 7 for cardiac complications and 1 for cancer. Thirty-six patients (13.5%) had hospital readmission for cardiac causes (mostly for atrial fibrillation or other arrhythmias (9.3%), but none needed cardiac surgical reintervention; 21 patients (7.9%) were hospitalised for non-cardiac causes. No limitation in function activities of daily living was reported by most patients (94%), 92% perceived their general health as excellent, very good or good and none considered it insufficient; 80% were NYHA I, 17% NYHA II, 3% NYHA III and none NYHA IV. Multivariate analysis indicated preoperative treatment with digitalis or nitrates, and postoperative cardiac biomarkers release was independently associated to death. Elevated cardiac biomarker release and length of hospital stay were the only postoperative independent predictors of death in this study. PMID- 19136753 TI - Butorphanol premedication to facilitate invasive monitoring in cardiac surgery patients before induction of anaesthesia. AB - Cannulations (peripheral vein, radial artery and jugular vein) performed for invasive monitoring before induction of anaesthesia in cardiac surgery patients may be associated with stress and anxiety. The efficacy and safety of butorphanol premedication was assessed in setting up of invasive monitoring. The study was a prospective, randomized, double blind, placebo controlled one with 70 patients undergoing elective cardiac surgery. In group-1 patients (n = 35) (placebo) intramuscular saline was administered 1-2 hours before the surgery in equivalent volume to butorphanol. In group-2 (n = 35) butorphanol (1, 1.5 and 2 mg for three body weight groups < 40 kg, 41-60 kg and> 60 kg, respectively) was administered 1 2 hours before surgery. Observer blinded for medication recorded the sedation score, pupil size and pain after each cannulation using visual analogue score (VAS). Student's 't' test and Chi-square test for proportions, Mann-Whitney test for non-parametric data was carried out. The median pain score of cannulation in group-2 (butorphanol) in the hand (10 mm) and neck (20 mm) were significantly (P < 0.05) lower than group-1 (placebo) patients (hand = 30 mm and neck = 40 mm). Pain during neck cannulation was significantly (P < 0.05) reduced (VAS < 30 mm) in patients with the pupil size of < 2.5 mm. Since the pain during neck cannulation was more than pain during hand cannulations in both the groups, we conclude that the intensity of pain depends also upon the site of cannulation. Besides the analgesic effect of butorphanol, its sedative effect helped to effectively decrease the pain during neck cannulation in conscious patients. PMID- 19136752 TI - Caudal epidural sufentanil and bupivacaine decreases stress response in paediatric cardiac surgery. AB - Surgery and anaesthesia are known to cause stress response. Attenuation of stress response can decrease morbidity, postoperative hospital length of stay and, thus, cost. Intrathecal and epidural techniques produce reliable analgesia in patients undergoing surgery along with stress response attenuation. The present study was undertaken to evaluate the efficacy of caudal sufentanil and bupivacaine combination on perioperative stress response in paediatric patients undergoing open heart surgery. Thirty patients (ASA grade II-III) undergoing elective corrective cardiac surgery for acyanotic congenital heart disease, were randomly allocated to two groups. In group GA (n = 15), patients received balanced general anaesthesia. In group GC (n = 15), in addition to general anaesthesia, caudal block with bupivacaine and sufentanil combination was given after endotracheal intubation. Monitoring included electrocardiography, invasive arterial pressure, end-tidal carbon dioxide, pulse oximetry, arterial blood gases including serum electrolytes, blood glucose, serum cortisol, urine output, central venous pressure and temperature. Haemodynamic responses in both groups were statistically similar. Serum cortisol levels were significantly lower in GC group than GA group (P < 0.05) after sternotomy (9.8+/-7.5 vs. 34.74+/-27.35), on cardiopulmonary bypass (CPB) (12.17 +/- 6.2 vs. 35.36 +/- 24.15), after sternal closure (14.03 +/- 5.1 vs. 37.62 +/- 20.69), 4 hours (26.64 +/- 14.61 vs. 37.62 +/- 9.13) and 24 hours (14.30 +/- 8.11 vs. 28.12 +/- 16.31) after intubation. Blood glucose levels were significantly higher in GA group as compared to GC group at sternal closure (277.46 +/- 77.25 vs.197.73 +/- 42.17) and 4 hours (255.26 +/- 73.73 vs. 185.26 +/- 57.41) after intubation (P < 0.05). To conclude, supplementation of caudal epidural bupivacaine and sufentanil could effectively attenuate the stress response in paediatric patients undergoing cardiac surgery under CPB in acyanotic congenital heart anomaly. PMID- 19136754 TI - Surgical retrieval of embolised atrial septal occluder device from pulmonary artery: pathophysiology and role of the intraoperative transoesophageal echocardiography. AB - Atrial septal defect is usually closed in the cardiac catheterisation laboratory using atrial septal occluder (ASO) device. One of the complications associated with the procedure is embolisation of the device into the pulmonary artery. We are reporting two cases wherein the pulmonary embolisation of ASO device occurred during the procedure in one patient and in the early post-procedure period in another; both were retrieved surgically. We are also describing the haemodynamic consequences of this complication and the role of intraoperative transoesophageal echocardiography during surgical retrieval of the device. PMID- 19136755 TI - Induced mild hypothermia in post-cardiopulmonary bypass vasoplegia syndrome. AB - The state of vasoplegia in immediate post-cardiopulmonary bypass period is characterized by severe hypotension, supranormal cardiac output, low systemic vascular resistance (SVR), and resistance to vasoconstrictors. We could successfully use induced mild hypothermia to increase SVR, and could avoid very high doses of nor-epinephrine (>0.3 mcg/kg/min) in the background of severe pulmonary hypertension (systolic pulmonary pressure> 90 mmHg). Its effects such as decreased oxygen demand, positive inotropy and better right ventricle performance probably helped to improve oxygenation in presence of pulmonary oedema. PMID- 19136756 TI - Superior vena cava syndrome after pulsatile bidirectional Glenn shunt procedure: perioperative implications. AB - Bidirectional superior cavopulmonary shunt (bidirectional Glenn shunt) is generally performed in many congenital cardiac anomalies where complete two ventricle circulations cannot be easily achieved. The advantages of BDG shunt are achieved by partially separating the pulmonary and systemic venous circuits, and include reduced ventricular preload and long-term preservation of myocardium. The benefits of additional pulsatile pulmonary blood flow include the potential growth of pulmonary arteries, possible improvement in arterial oxygen saturation, and possible prevention of development of pulmonary arteriovenous malformations. However, increase in the systemic venous pressure after BDG with additional pulsatile blood flow is known. We describe the peri-operative implications of severe flow reversal in the superior vena cava after pulsatile BDG shunt construction in a child who presented for surgical interruption of the main pulmonary artery. PMID- 19136757 TI - Intraoperative myocardial ischemia during renal transplantation caused by anomalous origin of the right coronary artery. AB - Anomalous origin of the right coronary artery (AORCA) is a rare congenital anomaly with an incidence of 0.92% during routine cardiac catheterization. Its presence raises an important concern to the anaesthesiologist because it can lead on to myocardial ischaemia manifesting as either angina pectoris or myocardial infarction, or sudden death in young patients with minimal exertion, even in the absence of atherosclerosis. Patients with AORCA may be intolerant to stress and the high cardiac output condition owing to volume loading. Such a therapeutic manoeuvre may be desirable during renal transplantation to enable better perfusion of the renal graft immediately after grafting the kidney, in order to improve its function. Hence, haemodynamic goals in renal transplant recipient with AORCA can be contradictory during surgery, thereby rendering anaesthetic management challenging. We report a case of acute myocardial ischemia precipitated by fluid loading conditions in a patient with AORCA during renal transplant that was successfully treated with emergent intra-aortic balloon pump therapy intraoperatively. Judicious intraoperative fluid replacement is recommended, and volume overload must be avoided in AORCA patients undergoing surgery. PMID- 19136758 TI - Anaesthetic management of a child presenting with intrapericardial teratoma compressing the airway and the heart. AB - Intrapericardial tumours can cause cardiac compromise either from pericardial effusion producing cardiac tamponade or from the mass obstructing the outflow from the heart or both. Respiratory embarrassment may also be present if the mass compresses the trachea or the main stem bronchus. A 1-month-old child presented in the emergency needing prompt measures to relieve compressive symptoms. Successful anaesthetic and surgical management led to uneventful recovery. Histopathology of the mass revealed a rarely occurring intrapericardial teratoma. After surgical removal, the child had clinical follow-up for more than a year. Intrapericardial teratoma is a rare entity that presents a diagnostic and therapeutic challenge. The authors describe the case of a 1-month-old male child with this condition who was successfully managed by resection of the mass. PMID- 19136759 TI - Milrinone infusion: a therapeutic option in coronary vasospasm after primary percutaneous transluminal coronary angioplasty. AB - A 42-year-old male presented to the emergency department with acute chest pain. The electrocardiogram revealed inferior wall myocardial infarction. Emergency coronary angiography revealed total occlusion of the distal right coronary artery with thrombus. Patient was taken up for primary percutaneous coronary angioplasty with stenting of distal right coronary artery. Six hours following the procedure, the patient developed re-elevation of ST-segment in inferior leads of electrocardiogram and subsequent haemodynamic instability. Repeat coronary angiography revealed patent stent and coronary artery spasm in proximal part, which was relieved by intracoronary injection of nitroglycerine. After an hour, the patient re-developed symptoms of chest pain along with bradycardia, hypotension and ST segment elevation. Intravenous infusion of nitroglycerine did not improve the condition but produced persistent hypotension. Infusion of milrinone was then started. Over time, normalisation of electrocardiogram occurred. The patient was discharged in stable condition. This case suggests that milrinone may be effective in alleviating coronary artery spasm when the use of other agents fails. PMID- 19136760 TI - Adult cardiac transplantation: a review of perioperative management Part-I. AB - Cardiac allotransplantation has, over the years, become the established therapeutic modality for patients with end-stage heart failure. Significant advances in immunosuppressive therapy have dramatically improved the outcome of heart transplantation over the past four decades. This review will focus on the anaesthetic challenges involved in the perioperative management of these complex patients as well as some of the proposed alternatives to transplantation. PMID- 19136761 TI - Intramyocardial dissecting haematoma causing cardiac tamponade: an unusual complication after mitral valve replacement surgery. PMID- 19136762 TI - Large rhabdomyosarcoma of the right ventricle obstructing tricuspid valve, pulmonary valve and left ventricular outflow tract. PMID- 19136763 TI - Selective lung separation with Fogarty catheter guided by an adult fiberoptic bronchoscope in a paediatric patient with situs inversus. PMID- 19136764 TI - Intraoperative coronary vein graft thrombosis: should we blame the use of aprotinin? PMID- 19136765 TI - The value of postoperative troponin levels in predicting long-term mortality after coronary artery bypass surgery. PMID- 19136766 TI - Identical drug packaging: heparin and midazolam-yet another instance of similar drug packaging. PMID- 19136767 TI - Diagnostic fiberoptic bronchoscope aided double lumen tube insertion. PMID- 19136768 TI - Quality control in the histopathology laboratory: an overview with stress on the need for a structured national external quality assessment scheme. AB - The concept of quality control in histopathology is relatively young and less well understood. Like in other disciplines of laboratory medicine, the concept of quality and its control is applicable to pre analytical, analytical and post analytical activities. Assessment of both precision and accuracy performances is possible by appropriate internal and external quality control and assessment schemes. This article is a review of all processes that achieve quality reporting in histopathology. There is a special focus on external quality assessment - a scheme that lacks organization on a national level in our country. Statistical data derived from a small scale external quality assurance program is also analyzed along with recommendations to organize an effective national scheme with the participation of authorized zonal centers. PMID- 19136769 TI - CD20 positivity in classical Hodgkin's lymphoma: Diagnostic challenge or targeting opportunity. AB - BACKGROUND: It is now well established that Hodgkin cells are clonal B cells with a CD30 and CD15 phenotype. However, on immunohistochemistry, in our experience and the experience of others, CD20 positivity in an otherwise typical classical Hodgkin's Lymphoma is not uncommon and if associated with CD15 negativity poses a potential diagnostic trap and is likely to be called B-NHL. OBJECTIVE: To assess the frequency of B-cell related antigens CD20 and CD79a in classical Hodgkin's Lymphoma. MATERIALS AND METHODS: A total of 91 consecutive cases of classical Hodgkin's Lymphoma were analyzed for co-expression of CD20 and CD79a. Both males and females of all ages were included in this study. All cases of nodular lymphocyte predominant Hodgkin's Lymphoma were excluded. All the cases were stained with a panel of antibodies including LCA, CD20, CD79a, CD30, CD15, CD3, EMA and Alk. Protein. RESULTS: All 91 cases of classical Hodgkin's Lymphoma showed negativity for LCA and positivity for CD30. Eighteen cases (19.8%) showed distinct membrane staining with CD20 in most of the large atypical cells. However, out of these, only 7 cases (7.7%) showed CD79a co-expression, which was largely focal. CD15 negativity with CD20 positivity was seen in 7 (7.7%) cases of otherwise typical classical Hodgkin's Lymphoma. CONCLUSIONS/RECOMMENDATIONS: CD20 expression is frequent in classical Hodgkin's Lymphoma and our results are in consensus with reported literature on this subject. In these cases, LCA negativity of large cells was extremely useful in clinching the right diagnosis. PMID- 19136770 TI - Liver needle biopsy in Iraninan pediatric patients: diagnostic significance and pattern of liver diseases. AB - We aimed at determining the pattern of liver disease in the Iranian children referred to the Medical Center of Children affiliated with the Tehran University of Medical Sciences. MATERIALS AND METHODS: In a cross-sectional study conducted over 2 years, 425 liver needle biopsies were sent to the pathology laboratory of our center. Slides were prepared from paraffin-embedded blocks, stained by routine H & E and special stains and were then reviewed. The frequency of each disorder, separately and in combination with the age group or gender of the patients was calculated and compared with other similar studies. RESULTS: The male to female ratio was 1.42:1. The age range was between 1 month and 18 years old and 41.4% were less than 2 years old. The most common histological diagnosis was iron overload due to major thalassemia (17.5%) followed by biliary atresia (9.7%), no significant pathologic change (8.7%), neonatal hepatitis (8.7%), chronic hepatitis (8.5%), cirrhosis (6.5%), metabolic disease (5.5%) and progressive familial intrahepatic cholestasis (5%). RESULTS of the hemosiderosis grading in patients with thalassemia revealed no or minimal, mild, medium, or marked increase in 10%, 27.1%, 10%, 21.4% and 31.5% of the cases, respectively and the degree of iron deposition rose in parallel with age and also the stage of fibrosis (p< 0.05). CONCLUSION: A liver biopsy is a useful and practical tool for the appropriate diagnosis of pediatric liver diseases. Also, we found that in non thalassemic children, biliary atresia, chronic hepatitis and neonatal hepatitis, in the stated order, are the most prevalent histologic diagnoses in Iranian pediatrics. PMID- 19136771 TI - IgA nephropathy in Kerala, India: a retrospective study. AB - This study was conducted to assess the prevalence of IgA nephropathy in Kerala. No published data are available in literature from this part of India. This study included 1592 renal biopsies received from all parts of Kerala over a 2-year period. The age of the subjects ranged from 5 to 78 years old with a peak incidence in the third decade of life. The male:female ratio was 1.2:1. A total of 60% of the cases had presented with hematuria. Direct immunoflourescence was performed using specific antibodies for IgA, IgG, IgM, C3 and C1q. Light microscopic studies with special stains namely PAS, silver and trichrome were also done. A total of 227 cases (14.26% of all renal biopsies) were diagnosed as IgA excluding 12 cases of Henoch-Schonlein Purpura (12.74% in the first year and 15.52% in the second year). The prevalence of IgA nephropathy appears to be increasing in Kerala. PMID- 19136772 TI - Seroprevalence of hepatitis viruses in patients infected with the human immunodeficiency virus. AB - OBJECTIVE: The co-infection of Hepatitis B and C viruses with HIV accelerates disease progression and also has an effect on the management of patients infected with HIV. The prevalence of HIV co-infection with hepatitis viruses varies widely. This study is planned to evaluate the prevalence of HIV co-infection with Hepatitis B and C viruses in North India. MATERIALS AND METHODS: A total of 1178 patients enrolled in the ART center were retrospectively analyzed for the presence of HBV and HCV on the basis of the presence of HBsAg and anti-HCV markers. RESULTS: In patients infected with HIV, the prevalence of co-infection with HBV was 9.9% (117/1178), the prevalence of co-infection with HCV was 6.3% (74/1178) and the prevalence co-infection with both HBV and HCV was ~1% (12/1178). DISCUSSION: The prevalence rate of HBV and HCV are increasing in patients infected with HIV. Having acquired the knowledge about the importance of such a co-infection, it is essential that all the patients infected with HIV be screened for HBV and HCV co-infection. PMID- 19136773 TI - Evaluation of endoscopic biopsy in gastric lesions with a special reference to the significance of mast cell density. AB - Various studies have shown the role of mast cells in chronic inflammatory states and in tumor growth. The study is designed to have an idea of the relationship of mast cell density (MCD) to gastric ulcer and cancer, to verify whether mast cell accumulation occurred in the two conditions especially in Indian patients and thus postulate that therapeutic strategies against mast cell mediators could be useful in treatment. Also, we want to review literature and attempt to explain our findings. A total of 240 patients, who underwent their first endoscopy and biopsy for a span of 21/2 years were studied retrospectively. Out of these, 210 cases that were either gastric ulcers or cancer were chosen for this MCD study. Biopsies were sectioned and stained routinely. Toluidine blue stain and copper grid was used to calculate MCD. Student's t-Test was used to calculate the statistical significance of MCD. MCD in benign ulcers was much higher than in control subjects. MCD in well-differentiated cancers showed MCD higher than control. Poorly-differentiated adenocarcinoma showed lower MCD than well differentiated adenocarcinoma. It was concluded that the accumulation of mast cells in gastric ulcers is an inflammatory response. MCD is increased in well differentiated gastric cancers, which may be a mast cell mediated immune response or mast cells may have a role in tumor angiogenesis and produce factors for tumor progression. Poorly-differentiated adenocarcinoma apparently lacks mast cell mediated anti-tumor response in some unexplained way. PMID- 19136774 TI - Utility of fibronectin in immuocytochemial differentiation of reactive mesothelial cells from metastatic malignant cells in serous effusions. AB - Cytological evaluation of serous effusions often poses difficulties to the pathologists. This study was designed to assess the utility of fibronectin as a mesothelial cell marker and evaluate its use along with carcinoembryonic antigen (CEA) as a short panel to aid in the differentiation of reactive mesothelial cells from metastatic adenocarcinoma cells in serous effusions. Forty serous effusion cases from clinically benign cases and forty from clinically malignant cases with a known primary were included in the study. After routine cytological evaluation, the cases were categorized as benign, suspicious and malignant. All the cases were studied for fibronectin and CEA immunostaining using APAAP technique. In the present study, fibronectin emerged as a 100% specific and 93.4% sensitive marker for mesothelial cells. CEA immunostaining was seen in 80% cases of metastatic adenocarcinoma in serous effusions. Mesothelial cells did not show any CEA positivity. Short panel of CEA and fibronectin aided in reaching a correct diagnosis in three out of five cytologically suspicious cases. PMID- 19136775 TI - Morphological spectrum of peripheral nerve sheath tumors: a series of 126 cases. AB - By convention, soft tissue tumors include the peripheral nerve sheath tumor (PNST) although they arise from neuroectoderm. PNSTs display a wide spectrum of morphology and biological behavior. Malignant transformation in benign tumors, especially neurofibroma, is an acknowledged phenomenon.This article describes clinico-pathological features of 126 PNSTs with their morphological variants and those associated with neurofibromatosis 1 (NF1) encountered in a single center over a 6-year period. Benign PNSTs were more common [118 (93.65%)] when compared with their malignant counterparts [8 (6.34%)]. Histological variants including plexiform schwannomas and diffuse, plexiform and pigmented forms of neurofibromas, though rare, were observed. Eleven (8.73%) patients with NF1 showed a strong association with plexiform neurofibroma. A malignant transformation in pre-existing neurofibroma was seen in 1 patient with NF1. This series highlights the clinico-pathological spectrum of PNSTs, their morphological variants and the incidence of associated NF1. The importance of detecting plexiform variants lies in their vastly different prognostic implications particularly when occurring in the setting of NF1. PMID- 19136776 TI - A comparative study of cervical smears in an urban Hospital in India and a population-based screening program in Mauritius. AB - OBJECTIVE: To study cervical smear abnormalities in urban women in India and women in Mauritius and to compare the results in the two groups. STUDY DESIGN: An analysis of 6010 cervical smears taken as part of routine check-ups in an urban hospital was done and an analysis of 10,000 cervical smears taken from women participating in a National Cancer Screening Program in Mauritius was done. Emphasis was put on cervical epithelial cell abnormalities and the results in the two populations are compared with that of similar studies in other parts of the world. RESULTS: Non specific inflammation formed 19.6% and 25.34% of the smears in the Indian and Mauritian groups, respectively (with specific infection forming 6.05% and 15.08%). The epithelial abnormalities constituted 1.392% of the Indian group and 0.47% of the Mauritian group. The difference was statistically significant in the atypical squamous cells of uncertain significance (ASCUS) and atypical glandular cells of uncertain significance (AGUS) group. CONCLUSIONS: The prevalence of low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL) is similar to that in the developed world. PMID- 19136777 TI - PNH revisited: Clinical profile, laboratory diagnosis and follow-up. AB - BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by intravascular hemolysis, marrow failure, nocturnal hemoglobinuria and thrombophila. This acquired disease caused by a deficiency of glycosylphosphatidylinositol (GPI) anchored proteins on the hematopoietic cells is uncommon in the Indian population. MATERIALS AND METHODS: Data of patients diagnosed with PNH in the past 1 year were collected. Clinical data (age, gender, various presenting symptoms), treatment information and follow-up data were collected from medical records. Results of relevant diagnostic tests were documented i.e., urine analysis, Ham's test, sucrose lysis test and sephacryl gel card test (GCT) for CD55 and CD59. RESULTS: A total of 5 patients were diagnosed with PNH in the past 1 year. Presenting symptoms were hemolytic anemia (n=4) and bone marrow failure (n=1). A GCT detected CD59 deficiency in all erythrocytes in 4 patients and CD55 deficiency in 2 patients. A weak positive PNH test for CD59 was seen in 1 patient and a weak positive PNH test for CD55 was seen in 3 patients. All patients were negative by sucrose lysis test. Ham's test was positive in two cases. Patients were treated with prednisolone and/or androgen and 1 patient with aplastic anemia was also given antithymocyte globulin. A total of 4 patients responded with a partial recovery of hematopoiesis and 1 patient showed no recovery. None of the patients received a bone marrow transplant. CONCLUSION: The study highlights the diagnostic methods and treatment protocols undertaken to evaluate the PNH clone in a developing country where advanced methods like flowcytometry immunophenotyping (FCMI) and bone marrow transplants are not routinely available. PMID- 19136778 TI - The absence of JC virus antigens in Indian children with medulloblastomas. AB - BACKGROUND: The human polyoma virus, also known as the JC virus (JCV), replicates predominantly in the oligodendrocytes, the myelin producing cells in the central nervous system and results in the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML) especially in immunosuppressed patients with AIDS. Several investigators have also documented the presence of the viral genome and early and late antigens in a variety of brain tumors particularly in medulloblastomas, gliomas and ependymomas. Reports also indicate the presence of JCV in patients with colon cancer. The T antigen of JCV has been postulated to have oncogenic potential as substantiated by animal experiments. Although JCV infects 80% of the population, there are scant epidemiological studies regarding JCV from India. There are also reports of the low prevalence of PML in patients with AIDS from India and Africa. AIM: This study was undertaken to investigate if Indian children with medulloblastomas also show evidence of JCV. METHODS: Twenty two consecutive cases of medulloblastomas were investigated for the presence of T antigen and agnoprotein of JCV in biopsy specimens by immunohistochemistry. Antibodies to the agnoprotein antigen raised in rabbits and a monoclonal antibody against SV40 T antigen raised in mice that cross-reacts with JCV T antigen were used. RESULTS: Out of 22 patients, 4 had desmoplastic tumors while the rest had classical tumors. All children were below the age of 10. Results indicate that while PML tissues showed consistent immunostaining both with antibody to T antigen and agnoprotein antibody, none of the tumors showed any positive staining for JC viral antigens. CONCLUSION: JCV antigens could not be detected by immunohistochemistry in the tumor tissues of Indian children with medulloblastomas. PMID- 19136779 TI - Congenital rubella and cytomegalovirus infections in and around Chandigarh. AB - AIMS: This study has analyzed the role of rubella and cytomegalovirus (CMV) in infections of children and pregnant women. SETTINGS AND DESIGN: The study was carried out in a tertiary care hospital. Data from blood samples from pregnant women (asymptomatic and also women with obstetric problems) and children (suspected of intrauterine infections) that were received in the laboratory over a period of 8 years were analysed. MATERIALS AND METHODS: The samples were tested for rubella- and CMV-specific IgM antibodies by capture enzyme linked immunosorbent assay. RESULTS: In children, the overall positivity for rubella- and CMV-specific IgM antibodies was 2.8% and 12.5%, respectively. In asymptomatic pregnant females, rubella positivity was 0.7% while in women with obstetric complications it was 3.4%. IgM antibody positivity in cases of CMV was 7.8% in both asymptomatic pregnant women and also in women with obstetric complications. CONCLUSIONS: The study indicated that infection with CMV is more common than the rubella virus. The incidence of rubella has reduced over the past few years. Hence, screening for rubella infection may be reserved for women with obstetric complications only. The routine screening for CMV among all antenatal cases is a debatable issue. PMID- 19136781 TI - Trends in antimicrobial resistance of fecal Shigella and Salmonella isolates in Tehran, Iran. AB - CONTEXT: The resistance of bacteria to commonly prescribed antibiotics is increasing both in developing as well as developed countries. Resistance has emerged even to newer, more potent antimicrobial agents. The present study was therefore undertaken to report resistance rates to antimicrobial agents in 2487 stool culture isolates in a tertiary care hospital between 1996 and 2000 and 2001 and 2005. MATERIALS AND METHODS: During 1996 to 2005, 31776 fecal samples were collected from all patients having diarrhea aged> 1 month to 14 years old. Microbiology records were reviewed and information on each isolate regarding its antimicrobial susceptibility profile was collected and recorded. STATISTICAL ANALYSIS USED: The statistical analysis was performed using SPSS, Version 11.5 software. RESULTS: Of the positive fecal cultures, 1329 (53.43%) of the isolates were Shigella spp and 700 (28.14%) of the isolates were Salmonella spp. Resistance to antimicrobial agents increased among most of the pathogens between 2001 and 2005. An increase in the rate of resistance was observed in Shigella spp for kanamycin (from 11 to 37%) and ceftazidime (from 1 to 9.9%) and among Salmonella spp. for nalidixic acid (from 9.2 to 42.3%) and ceftazidime (from 3 to 23.4%). CONCLUSIONS: Routine surveillance of antimicrobial susceptibilities to all classes of clinically used agents is necessary to detect resistance trends in different parts of world, detecting the emergence of new resistance mechanisms that guide infection control measures and public health guidelines; such trends may help in identifying outbreaks of resistant organisms. Such a check seems to be the best way to find appropriate antibiotic regimens. PMID- 19136780 TI - Inducible clindamycin resistance among clinical isolates of Staphylococci. AB - INTRODUCTION: Clinical failure of clindamycin therapy has been reported due to multiple mechanisms that confer resistance to macrolide, lincosamide and streptogramin antibiotics. This study was undertaken to detect the presence of inducible clindamycin resistance among clinical isolates of staphylococci. MATERIALS AND METHODS: The detection of inducible clindamycin resistance was performed by D-test using erythromycin and clindamycin discs as per CDC guidelines. RESULTS: Among the 244 clinical isolates of staphylococci studied, 32 (13.1%) showed inducible clindamycin resistance and belonged to the MLSBi phenotype. Among the MLS B i phenotypes, 10 isolates were methicillin-resistant Staphylococcus aureus (38.4% of the total MRSA), 16 were methicillin-sensitive Staphylococcus aureus (12.9% of the total MSSA) and 6 were coagulase-negative staphylococci (6.3% of the total CONS). CONCLUSION: The test for inducible resistance to clindamycin should be included in the routine antibiotic susceptibility testing, as it will help in guiding therapy. PMID- 19136782 TI - Bacterial vaginosis with special reference to anaerobes. AB - AIMS: This study was undertaken to assess the prevalence of bacterial vaginosis (BV) and to estimate the prevalence of anaerobic organisms in vaginal discharge of women suffering from bacterial vaginosis. SETTINGS AND DESIGN: Patients attending the Obstetrics and Gynecology Department of a Medical College Hospital. A one year cross-sectional study. METHODS AND MATERIALS: High vaginal swabs taken from 174 female patients complaining of abnormal vaginal discharge. BV was diagnosed by clinical composite criteria and by gram stain. Anaerobes were isolated and identified from the discharge. STATISTICAL ANALYSIS USED: Chi square test, with level of significance set at a value of P< 0.05. RESULTS: BV was diagnosed in 68.39% of the cases by using clinical composite criteria and in 58.4% of the cases by gram stain. Anaerobic culture isolation of vaginal swabs revealed that out of 174 cases 143 (82.65%) were culture positive for anaerobes. Bacteroides were significantly raised in BV as compared with non bacterial vaginosis (NBV; < 0.05%). CONCLUSIONS: Anaerobic bacteria are important pathogens in the causation of bacterial vaginosis along with other aerobic organisms. Bacteroides and peptostreptococci are significantly raised in BV. PMID- 19136783 TI - Transport and storage of sputum specimen by using cetylpyridinium chloride for isolation of mycobacteria. AB - Of the 191 sputum specimens that were collected from pulmonary tuberculosis patients, 78.65% (140/178) specimens were culture positive when processed within 48 h by the NaOH method. The culture positivity in the same specimen that were stored with cetylpyridinium chloride (CPC) and processed after 7-8 days was 70.22% (125/178), whereas those stored without CPC and processed by the NaOH method was 46.62% (83/178). The difference in number of positive cultures obtained before storage and after storage (without CPC) was statistically significant (P = 0.001). Culture positivity by the CPC method was comparable with that of NaOH method before storage and the difference was not statistically significant (P = 0.35). PMID- 19136784 TI - Primary laryngeal leishmaniasis: a rare case report. AB - Laryngeal leishmaniasis is extremely rare. We report a case of primary laryngeal leishmaniasis in a 70-year-old male who was admitted with complaints of gradual progressive hoarseness of the voice, dyspnea, cough for the past 3 months and noisy breathing for the past 5 days. An X-ray of the soft tissue of the neck showed a prevertebral soft tissue mass causing narrowing of the airway at the C6 C7 vertebral level. A computerized tomography (CT) scan showed a soft tissue mass in the subglottic region causing significant narrowing of the airway. A direct laryngoscopy showed a pinkish-white, friable mass involving the subglottic region and the anterior half of the vocal cords. With the clinical suspicion of malignancy, an endoscopic biopsy was done. A histopathological examination showed diffuse mixed inflammatory cell infiltrate in subepithelium with numerous Leishmania donovani bodies in the cytoplasm of histiocytes. PMID- 19136785 TI - Primary langerhans cell histiocytosis of the vulva: report of a case and brief review of the literature. AB - Primary Langerhans cell histiocytosis (LCH) of the vulva is rare. Fifteen cases of primary cutaneous vulvar LCH have been reported in English literature. We report an additional case of LCH confined to the vulva. In this article, we describe the clinical presentation, histopathology and immunohistochemistry findings of vulvar LCH that are helpful to both gynecologists and pathologists in the diagnosis of this entity. We briefly discuss the pathogenesis of LCH. The debate whether LCH is a reactive or neoplastic entity is still ongoing. PMID- 19136786 TI - Anaplastic large-cell lymphoma with florid granulomatous reaction: a case report and review of literature. AB - Granulomatous reactions have been reported in association with lymphomas, more often with Hodgkins disease than with Non-Hodgkins Lymphoma. Not many reports are available on the association of anaplastic large-cell lymphoma with sarcoid-type granuloma. Herein, we report a case of an elderly female with generalized lymphadenopathy who had a florid granulomatous reaction almost masking the lymphoma cells in the lymph node biopsy. A detailed clinical history, careful histological examination and immunohistochemistry helped in attaining the correct diagnosis. PMID- 19136787 TI - Primary neuroendocrine carcinoma of the breast. AB - Primary neuroendocrine carcinoma of the breast is rare-only about 30 cases have been reported in literature. Immunohistochemical examination showing expression of chromogranin and/or synaptophysin confirms evidence of neuroendocrine differentiation. Usually foci of neuroendocrine differentiation can be seen in breast carcinoma and are reported to be present in about 2-5% of breast cancer cases. Here, we report a case of breast carcinoma in which most of the areas studied on the tissue section showed neuroendocrine differentiation. PMID- 19136788 TI - Malignant peripheral nerve sheath tumor with divergent differentiation. AB - A malignant peripheral nerve sheath tumor (MPNST) is an uncommon spindle cell sarcoma accounting for approximately 5% of all soft tissue sarcomas. A 55-year old female with a right suprarenal tumor showed MPNST with additional foci of epithelioid, rhabdomyoblastic, osteogenic and lipogenic differentiation. Although the capacity of MPNST to undergo epithelioid, rhabdomyoblastic, osteogenic and very rarely lipogenic differentiation is reported in literature, the occurrence of all these differentiation in one case has not been described in literature before. To the best of our knowledge, this is only the second MPNST case with lipomatous differentiation. PMID- 19136789 TI - Glomus tumor of the stomach. AB - Gastric glomus tumors are rare neoplasms. We report here a case of gastric glomus tumor in a 25-year-old female who presented with exsanguinating gastrointestinal hemorrhage. Clinically and on gross examination, the tumor was suspected to be a gastrointestinal stromal tumor (GIST). Histopathological and immunohistochemical evaluation revealed it to be a glomus tumor. PMID- 19136790 TI - Primary osteoliposarcoma of the bone. AB - Osteoliposarcoma are rare tumors of the bone. To our knowledge, so far only eight cases have been reported. Some of the reports have referred to these tumors as malignant mesenchymoma. We report a case of a male with osteoliposarcoma of the right femur who presented with pain and restricted movements of the right knee joint. Radiologically, a tumor in the lower end of the right femur was seen extending into the soft tissue. The biopsy was reported as osteosarcoma. The patient received three cycles of neo-adjuvant chemotherapy followed by limb salvage surgery with provisions for a custom-made prosthesis. A histopathological study of the excision specimen revealed areas of pleomorphic liposarcoma with numerous osteoblasts associated with areas of osteoid surrounded by neoplastic cells. The final diagnosis was osteoliposarcoma. Only 21% tumor necrosis (effects of chemotherapy) was observed. Presently, 26 months following diagnosis, the patient is fine with no evidence of local recurrence or distant metastasis. PMID- 19136791 TI - Secretory carcinoma arising in radial scars of the breast: A case report and review of literature. AB - Radial scars or complex sclerosing lesions are common benign lesions in the breast with characteristic radiological and pathological features. The pathological diagnosis of carcinoma arising in this setting requires careful amalgamation of clinical, radiological and morphological details. Ancillary techniques like immunohistochemistry aid in the diagnosis. We report an unusual case of a secretory carcinoma arising in the background of a radial scar. PMID- 19136792 TI - Choristoma of the base of the tongue. AB - Choristoma is a benign tumor where new bone formation occurs. It occurs exclusively in the flat bones of the skull and face. These are slow growing lesions that are usually completely asymptomatic and only present when there is a disruption in the function of the organ due to its large size as it grows. These choristomas can rarely occur in soft tissues especially in the head, eye, tongue, or extremities. Choristomas of the soft tissues are very rare. Only 61 cases of choristomas of the tongue have been reported in literature. Here we report a case of choristoma in the base of the tongue in a 25-year-old Malay female. PMID- 19136793 TI - Extra genital heterologous malignant mixed mullerian tumor of primary peritoneal origin. AB - The case of an extragenital heterologous malignant mixed mullerian tumor (MMMT) of primary peritoneal origin occurring in a 76-year-old female is presented. A large tumor was seen between the uterus and rectosigmoid occupying the entire pelvis. The uterus, fallopian tubes and ovaries were uninvolved. The tumor was composed of carcinomatous areas showing endometrioid and serous papillary differentiation and sarcomatous areas showing cartilaginous differentiation. The extragenital primary MMMTs of the female peritoneum are thought to originate from the secondary mullerian system. This case is presented for its rarity. To the best our knowledge, this is the first case of extragenital MMMT of primary peritoneal origin in Indian literature. PMID- 19136794 TI - Primary high-grade testicular leiomyosarcoma. AB - We herein present an extremely rare occurrence of primary intratesticular leiomyosarcoma. A 65-year-old patient presented with painless enlargement of the right testis. A high inguinal orchiectomy was done. Histopathological examination of the excised mass was consistent with high-grade leiomyosarcoma. Pertinent literature is reviewed and the importance of excluding the germ cell tumor and the paratesticular neoplasm is emphasized. PMID- 19136796 TI - Bilateral Sertoli-Leydig cell tumor of the ovary: a rare case report. AB - Sertoli leydig cell tumors also known as arrhenoblastoma, are a rare member of the sex cord-stromal tumor group of ovarian and testicular cancers, comprising less than 1% of all ovarian tumors, which occur in young adults and are almost always unilateral. We hereby report a case of a 17-year-old female presenting with a short history of irregular menses and an abdominal lump, which was histologically proven to be a bilateral sertoli leydig cell tumor of the ovary, an exceptionally rare entity in itself. PMID- 19136795 TI - Bilateral benign non functional struma ovarii with Pseudo-Meigs' syndrome. AB - Bilateral presentation of benign Struma ovarii is rare and has not been reported frequently in published literature. A 70-year-old postmenopausal female presented with progressive ascites, bilateral pleural effusion and elevated CA-125 levels. The contrast-enhanced computed tomography (CECT) of the abdomen and pelvis revealed a heterogenous mass in the left adnexa. These findings were suspicious for an ovarian malignancy. After surgery the diagnosis of non functional, bilateral benign Struma ovarii was made. Struma ovarii is a specialized ovarian teratoma composed predominantly of mature thyroid tissue. It is associated with pleural effusion and ascites (Pseudo-Meigs' syndrome) in 5% of cases. The combination of struma ovarii and elevated CA-125 levels has been reported infrequently. This is a rare case of bilateral benign struma ovarii associated with Pseudo-Meigs' syndrome and elevated CA-125 levels. Surgical excision of the ovarian masses induced immediate resolution of the ascites and pleural effusion and a reduction of the serum CA-125 level. PMID- 19136797 TI - Myoepithelioma of soft tissue. AB - Myoepitheliomas and mixed tumors involving deep subcutaneous and subfascial soft tissues of limb or limb girdle are rare lesions as against salivary lesions that are well established conditions. Here, we report a 22-year-old female who presented with painful hard swelling in the left gluteal region of 1(1/2) year duration. MRI showed a large ill-defined heterogeneous mass lesion measuring about 7-8 cm. in the left sacral region eroding the left sacroiliac region and left sacroiliac joint. With a clinical diagnosis of chondrosarcoma, the tumor with the surrounding tissue was resected in segments at surgery. Histomorphology revealed nests, sheets and cords of round to spindled cells with extensive squamous metaplasia in a myxoid to fibrous stroma. These cells extensively infiltrated muscle and bone. The tumor cells expressed immunoreactivity for cytokeratin (AE1/AE3) and S-100. PMID- 19136798 TI - Polymorphous low-grade adenocarcinoma of parotid gland: a rare occurrence. AB - Polymorphous low-grade adenocarcinoma (PLGA) is a rare salivary gland malignant tumor of low aggressiveness, commonly occurring in minor salivary glands. Its occurrence in major salivary gland has been documented albeit rarely. The striking histological feature is architectural diversity combined with benign cytologic features. We report a case of PLGA arising from left parotid in a 25 year-old male patient. On light microscopy, varied patterns were seen .The cells were uniform with bland nuclei. Neural invasion was noted. PMID- 19136799 TI - Reactive perforating collagenosis. AB - Reactive perforating collagenosis is a rare cutaneous disorder of unknown etiology. We hereby describe a case of acquired reactive perforating collagenosis in a patient of diabetes and chronic renal failure. PMID- 19136800 TI - Central neurocytoma in the vermis of the cerebellum. AB - In neuropathology, anatomic landmarks and topographic relationships bear particularly on issues of differential diagnosis. A case of central neurocytoma of the vermis of the cerebellum is being reported in this article. A 45-year-old male with features of hydrocephalous and a posterior fossa space-occupying lesion was diagnosed as having central neurocytoma upon a microscopy of Hematoxylene and Eosin stained sections. The diagnosis was confirmed by a positive immunohistochemical reaction for Neuron Specific Enolase (NSE) and Synaptophysin. This case report is being made to emphasize that a high index of suspicion is required even for reporting neuropathology where the site does not match the conventional location of the lesion seen and also to emphasize the importance of immunohistochemistry in proving such unusually located lesions. PMID- 19136801 TI - Hemoglobin E disorders in Eastern Uttar Pradesh. AB - The distribution of hemoglobin E (alpha2beta2 26Glu (R)Lys ) is mostly restricted to Northeastern India. While evaluating the patients of jaundice, we came across two cases of hemoglobin E (Hb E) disorders. The first case is in a 22-year-old Bengali male and the second case of Hb E/beta thalassemia in a 5-year-old Hindu boy. The family study revealed Hb E trait in both the parents of Case 1, whereas in Case 2, the father was found to have Hb E trait and the mother had beta thalassemia minor, thus confirming the diagnosis. Herein, we present the laboratory diagnosis and comparative data of the spectrum of Hb E disorders (i.e., heterozygous Hb E trait, homozygous Hb E disease and compound heterozygous Hb E/beta -thalassemia) that was found in our index cases and their parents. PMID- 19136802 TI - Transient thrombocytosis with megathrombocytes in a case of acute myeloblastic leukemia. AB - Thrombocytosis is commonly seen in reactive conditions and certain neoplastic states, such as chronic myeloproliferative disorders. It is rarely seen in acute leukemia. A 12-year-old girl with acute myeloblastic leukemia (FAB M2) in remission presented with pyoderma. Her hemogram revealed anemia (Hb-6.4g/dl), leucopenia (TLC - 1.2 x 109/L) and thrombocytosis (platelet count- 580 x 109/L). A peripheral blood film showed numerous abnormally large platelets with few atypical cells. The thrombocytosis subsided with the clearance of infection but atypical cells persisted. One month later, she relapsed. Cytogenetic analysis revealed variable results (trisomy 9 and deletion 3). This case has been presented because thrombocytosis is rare in AML and its appearance calls for a close follow-up. PMID- 19136803 TI - Osteomyelitis of the mandible due to Aggregatibacter (Actinobacillus) actinomycetemcomitans. AB - Aggregatibacter (Actinobacillus) actinomycetemcomitans is a capnoic gram negative coccobacilli known to produce juvenile periodontitis. This organism was isolated in pure culture from an unusual case of osteomyelitis of the mandible. The patient was treated with tetracycline, which is the drug of choice for A. actinomycetemcomitans and the clinical response improved. From our limited review of the literature, it appears that this is the first case of osteomyelitis due to A.actinomycetemcomitans reported in India. PMID- 19136804 TI - Isolation of Salmonella paratyphi A from renal abscess. AB - Intrarenal abscesses remain a significant cause of morbidity and mortality as well as a diagnostic dilemma because a plethora of microorganisms can cause this condition. A definitive diagnosis is made by demonstrating the organisms from the aspirate and the success or failure of therapy depends upon the antimicrobial sensitivity pattern. Enteric fever is a multisystem disorder caused by invasive strains of salmonella. Salmonellosis continues to be a major public health problem, especially in developing countries. Classic enteric fever is caused by S. typhi and usually less severe enteric fevers are caused by S. paratyphi A, B, or C. However, at times S. paratyphi is capable of causing serious and often life threatening infections like infective endocarditis, pericarditis, empyma, sino venous thrombosis, osteomyelitis, meningitis, bone marrow infiltration, hepatitis and pancreatitis. There are anecdotal case reports in world literature of abscesses being caused by this organism. Renal involvement like bacteriuria, nephrotic syndrome and acute renal failure have been reported due to S. parayphi A. S. paratyphi A has never been implicated in renal abscess, we report one such case that was managed successfully with medical therapy. PMID- 19136805 TI - Empyema thoracis due to actinomyces odontolyticus. AB - Actinomyces odontolyticus has been reported as an opportunistic pathogen causing systemic infections. A case of empyema thoracis due to this organism in a 68-year old male is reported here. The patient did not have any underlying disease or immunosuppression. The organism was isolated from his oral flora also. Eight cases of thoracopulmonary infections due to A. odontolyticus have been reported from the western countries, but none from India. PMID- 19136806 TI - Subperiodic, asymptomatic microfilaremia in an adult male from Mysore: a nonendemic area. AB - Wuchereria bancrofti is found throughout tropics and subtropics like Asia, Pacific islands, Africa, areas of South America and Caribbean basin. In all these areas, except Pacific islands, microfilaria occurs in the periodic form, in which case the microfilaria are found in large numbers in the peripheral blood during night. In the Pacific islands, they occur in the subperiodic form, i.e., microfilaria are present in the peripheral blood at all times and reach the maximum level of parasitemia in the afternoon. Microfilaria of Wuchereria bancrofti and Brugia malayi occurring in India displays a nocturnal periodicity, appearing in large numbers at night. This is the biological adaptation to the nocturnal biting habits of the vector mosquitoes. The maximum density in blood is reported between 10 PM and 2 AM. Here is a case report of asymptomatic microfilaremia showing subperiodicity, which is very unusual in India. PMID- 19136807 TI - Infection potpourri: are we watching? AB - Population explosion, frequent travel and urbanization have led to certain changes in the environment, which are conducive for the survival of multiple pathogens and their transmission vehicle in the same niche. Therefore, there are more chances of acquiring multiple infections at the same time with overlapping clinical manifestations. We would like to share a case having concurrent infection with four different agents. PMID- 19136808 TI - Siderophore production by uropathogenic Escherichia coli. AB - Urinary tract infection (UTI) is one of the most frequently encountered problems in ambulatory medicine. The present study was designed to determine siderophore production as the urovirulence factor of Escherichia coli isolated from the patients of UTI. A total of 160 strains of E. coli isolated from urine of patients with clinically diagnosed UTI were included in the study and 50 fecal isolates of E. coli, siderophore production was seen in 156 (97.5%). In 50 fecal isolates, siderophore production was seen in 2 (4%). Siderophore production has been shown to be more frequent in E. coli from patients with UTI, than in fecal isolates. The results suggest that siderophore production positive strains can be considered as UPEC. Thus, although a great deal has been learned regarding E. coli virulence mechanisms in UTI, much remains to be learned and the practical application of our growing understanding of E. coli virulence factors to the prevention and treatment of UTI has to be continued. PMID- 19136809 TI - Pearls from the abdomen. PMID- 19136811 TI - Ectopic adrenocortical rest in the wall of the large intestine. PMID- 19136810 TI - Primary mesenchymal chondrosarcoma of the breast. PMID- 19136812 TI - Primary peritoneal liposarcoma in a middle-aged patient. PMID- 19136813 TI - Group A Streptococcus meningitis: microbiological evidence clinches the issue. PMID- 19136815 TI - Proteomics in gerontology: current applications and future aspects--a mini review. AB - BACKGROUND: Aging is closely related to the onset of chronic diseases, such as coronary artery disease, diabetic nephropathy or different types of malignancies, reflecting the demand for novel biomarkers to manage theses diseases. OBJECTIVE: The analysis of the human proteome for biomarkers has made considerable advances in the last years. METHODS: We describe the main technological approaches taken, their advantages and disadvantages. RESULTS: We will review the different clinical sources of material and attempt to highlight the different challenges and approaches associated with these. Age-related changes in the proteome have been described and were found to be highly similar to changes associated with chronic diseases. We will give several examples on the successful application of proteomics in the diagnosis, prognosis and therapy of these chronic diseases. CONCLUSIONS: A boost in disease-related proteomic information is expected in the very near future, and will also result in its broad clinical application. However, this view appears to be dependent on the strict adherence to proper technological/analytical parameters, correct statistics, and large databases that allow comparison of datasets provided by different scientists. Clearly, the proteome is by far too complex to be tackled by one laboratory on its own. PMID- 19136814 TI - Sex differences in the cannabinoid modulation of appetite, body temperature and neurotransmission at POMC synapses. AB - We sought to determine whether sex differences exist for the cannabinoid modulation of appetite, body temperature and neurotransmission at pro opiomelanocortin (POMC) synapses. Gonadectomized male and female guinea pigs were outfitted to monitor core body temperature and injected with either the CB1 receptor agonist WIN 55,212-2 (1 mg/kg s.c.), antagonist AM251 (3 mg/kg s.c.) or vehicle (1 ml/kg s.c.) and evaluated for changes in six indices of feeding behavior under ad libitum conditions for 7 days. WIN 55,212-2 elicited an overt, sexually differentiated hyperphagia in which males displayed larger increases in hourly and daily intake, consumption/gram body weight, meal size and meal duration. The agonist also produced a more robust acute hypothermia in males than in females. In addition, males were more sensitive to the hypophagic effect of AM251, manifested by comparatively sizeable decreases in hourly intake, consumption/gram body weight, meal frequency and hyperthermia. To gain additional insight into the cellular mechanism underlying cannabinoid regulation of energy homeostasis, we performed whole-cell patch clamp recordings in hypothalamic slices prepared from gonadectomized male and female guinea pigs, and monitored miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs) in arcuate (ARC) neurons. ARC neurons from females exhibited a higher basal mEPSC frequency. WIN 55,212-2 dose-dependently reduced mEPSC and mIPSC frequency; however, cells from males were far less sensitive to the CB1 receptor-mediated decrease in mIPSC frequency. These effects were observed in neurons subsequently identified as POMC neurons. These data reveal pronounced sex differences in how cannabinoids influence the hypothalamic control of homeostasis. PMID- 19136816 TI - You never know what you might find--tracheo-esophageal fistula from extrusion of a spinal fusion fixation screw. PMID- 19136817 TI - Glomerular transcriptome changes associated with lipopolysaccharide-induced proteinuria. AB - BACKGROUND: Global gene expression patterns have recently been characterized in normal glomeruli, but gene expression changes that accompany glomerular disease remain poorly characterized. METHOD: Here, we mapped global glomerular gene expression profile changes occurring in conjunction with lipopolysaccharide (LPS) induced proteinuria in mice. RESULTS: We observed dramatic transcriptional reprogramming in glomeruli in response to LPS, representing some 20% of all genes and about 45% of the genes that are normally highly expressed in glomeruli. Bioinformatic analysis revealed significant changes in transcripts encoding proteins involved in the regulation of adherence junctions, actin cytoskeleton and survival in podocytes. In the LPS-treated mice, we observed dysregulation of genes expressed in glomerular endothelial and mesangial cells and in podocytes, there was also a significant decrease in podocyte number. Moreover, collagen alpha1, alpha2 (IV) and laminin 10 (laminin alpha 5 beta 1 gamma 1), which are expressed in immature glomeruli, were upregulated in the glomeruli of LPS-treated mice, suggesting remodeling of the glomerular basement membrane and activation of mesangial cells. By superimposing the LPS-induced changes onto GlomNet, a protein protein interaction network was predicted for podocyte proteins affected by LPS. CONCLUSIONS: The detected changes in glomerular gene expression and their involvement in protein interaction networks provide putative markers for early and transient glomerular injury and proteinuria. PMID- 19136818 TI - Association of serum total iron-binding capacity and its changes over time with nutritional and clinical outcomes in hemodialysis patients. AB - Serum transferrin, estimated by total iron-binding capacity (TIBC), may be a marker of protein-energy wasting (PEW) in maintenance hemodialysis (MHD) patients. We hypothesized that low TIBC or its fall over time is associated with poor clinical outcomes. In 807 MHD patients in a prospective 5-year cohort, associations of TIBC and its changes over time with outcomes were examined after adjustment for case-mix and markers of iron stores and malnutrition-inflammation including serum interleukin-6, iron and ferritin. Patients with serum TIBC >or=250 mg/dl had higher body mass index, triceps and biceps skinfolds and mid arm muscle circumference and higher serum levels of iron but lower ferritin and inflammatory markers. Some SF-36 quality of life (QoL) components were worse in the lowest and/or highest TIBC groups. Mortality was incrementally higher in lower TIBC levels (p-trend <0.001). Adjusted death hazard ratio was 1.75 (95% CI: 1.00-3.05, p = 0.05) for TIBC <150 compared to TIBC of 200-250 mg/dl. A fall in TIBC >20 mg/dl over 6 months was associated with a death hazard ratio of 1.57 (95% CI: 1.04-2.36, p = 0.03) compared to the stable TIBC group. Hence, low baseline serum TIBC is associated with iron deficiency, PEW, inflammation, poor QoL and mortality, and its decline over time is independently associated with increased death risk. PMID- 19136819 TI - Association between sweetened beverage consumption and body mass index, proportion of body fat and body fat distribution in Mexican adolescents. AB - BACKGROUND/AIMS: It was the aim of this study to evaluate the relationships between sweetened beverage (SB) consumption and the following indicators of overweight/fatness among Mexican adolescents: body mass index, body composition and body fat distribution. METHODS: We performed a cross-sectional analysis of data from adolescents participating in the baseline assessment of the Health Workers Cohort Study. Information on sociodemographic conditions, sexual maturation, dietary patterns and physical activity was collected via self administered questionnaires. SB consumption was evaluated through a validated semiquantitative food frequency questionnaire. Anthropometric measures were assessed with standardized procedures. The associations of interest were evaluated by means of multivariate regression and logistic regression models. RESULTS: A total of 1,055 adolescents, 10-19 years old (mean age 14.5+/-2.5 years), were evaluated. The overweight/obesity prevalence was 31.6% among girls and 31.9% among boys. We found that for each additional SB serving consumed daily, the subject's body mass index increased by on average 0.33 (p<0.001). Subjects consuming 3 daily servings of SB face a 2.1 times greater risk of proportionally excess body fat than those who consume less than 1 SB a day. CONCLUSIONS: Our data support the hypothesis that the consumption of SB increases the risk of overweight and/or obesity and encourages excess body fat and central obesity in Mexican adolescents. PMID- 19136820 TI - Ethanol consumption does not promote weight gain in female mice. AB - BACKGROUND: The prevalence of obese adult women has increased dramatically in the United States. Individuals consuming alcoholic beverages may obtain as much as 6 10% of their calories from ethanol; consequently, ethanol may contribute to a positive energy balance and weight gain in women consuming ethanol. The objective of these studies is to determine if ethanol consumption affects weight gain or body fat levels in female mice. METHODS: In order to determine the effects of ethanol consumption on weight gain, female mice were given either water or 20% w/v ethanol in the drinking water; mice were then placed on 1 of 3 diets for 20 weeks: (1) 30% calorie-restricted diet, (2) low-fat diet or (3) high-fat diet. Mice were scanned using a GE Lunar Piximus Densitometer to determine body fat, lean body mass and bone mineral density. RESULTS: Mice consuming the high-fat diet had the highest body weight. Moreover, ovariectomy exacerbated the effects of the high-fat diet. That is, ovariectomized female mice consuming the high-fat diet gained a higher amount of body weight and adipose tissue than non ovariectomized mice consuming the high-fat diet. Ethanol-consuming mice did not have a higher susceptibility to gaining body weight or body fat, even though they tended to have higher caloric intake than water-consuming mice. CONCLUSIONS: In female mice that consumed a high-fat diet, chronic ethanol consumption did not increase susceptibility to gaining weight or becoming obese. PMID- 19136821 TI - Effects of very-low-carbohydrate (horsemeat- or beef-based) diets and restricted feeding on weight gain, feed and energy efficiency, as well as serum levels of cholesterol, triacylglycerol, glucose, insulin and ketone bodies in adult rats. AB - BACKGROUND/AIMS: The beneficial or harmful effect of the low-carbohydrate (low carb), high-protein, high-fat diet (Atkins diet) has not been clearly demonstrated. We determined the effect of a low-carb diet and restricted feeding (70% ad libitum intake) on serum levels of cholesterol, triacylglycerol, glucose, ketone bodies and insulin in rats. METHODS: In experiment 1, each of 4 groups with 10 adult rats was assigned to a high-carb diet (AIN-93G) + ad libitum intake or restricted feeding, or a low-carb diet (53% horsemeat) + ad libitum intake or restricted feeding (2 x 2 factorial). In experiment 2, each of 3 groups with 10 adult rats was assigned to a control (AIN-93G) or low-carb diets (53% beef or horsemeat). RESULTS: Restricted feeding and the low-carb diet reduced (p<0.01) serum triacylglycerol compared with ad libitum intake and the AIN-93G diet, respectively (experiment 1). The dietary effect on serum total cholesterol, high density or low-density lipid cholesterol appeared to be inconsistent, but restricted feeding increased the low-density lipoprotein cholesterol level. The serum ketone body level was increased by the low-carb diet compared with AIN-93G (experiment 2). CONCLUSION: Restricted feeding and a low-carb diet are beneficial for alleviating cardiovascular disease risk factors, and their effects are additive, restricted feeding being more pronounced. PMID- 19136822 TI - Meta-analysis of enteral nutrition versus total parenteral nutrition in patients with severe acute pancreatitis. AB - OBJECTIVE: To compare the safety of enteral nutrition and total parenteral nutrition in nutrition support of patients with severe acute pancreatitis. METHODS: DATA SOURCES: Medline, Embase, and manual search. STUDY SELECTION: 295 articles were screened for randomized controlled studies (RCTs) that compared enteral nutrition with total parenteral nutrition in patients with severe acute pancreatitis. Finally, six RCTs were identified and included in the meta analysis. DATA EXTRACTION: six RCTs with 224 participants were analyzed. The main outcome were infections, artificial nutrition-related complications, pancreatitis related complications, non-pancreatitis-related complications, organ failure and mortality. The meta-analysis was performed with the fixed effects model or random effects model. RESULTS: Compared with total parenteral nutrition, enteral nutrition was associated with a significantly lower risk of infections [odds ratio (OR) 0.236; 95% confidence interval (95% CI) 0.120-0.464, p<0.001], pancreatitis-related complications (0.456; 0.234-0.888, p=0.021), organ failure (0.334; 0.167-0.670, p=0.002), multiple organ dysfunction syndrome (0.306; 0.128 0.736, p=0.008), and mortality (0.251; 0.095-0.666, p=0.005). There were no significant differences in artificial nutrition-related complications (0.642; 0.354-1.162, p=0.143), and non-pancreatitis-related complications (0.716; 0.325 1.576, p=0.406) between the two groups. CONCLUSIONS: Enteral nutrition appears safer than total parenteral nutrition in nutrition support of patients with severe acute pancreatitis. PMID- 19136823 TI - Human carotid plaque calcification and vulnerability. Relationship between degree of plaque calcification, fibrous cap inflammatory gene expression and symptomatology. AB - BACKGROUND: Inflammation is a key mechanism in human atherosclerotic plaque vulnerability and disruption. The objective was to determine the differential gene expression of pro- and anti-inflammatory factors in the fibrous cap and shoulder region of noncalcified and calcified carotid endarterectomy plaques. METHODS: Thirty carotid endarterectomy plaques were classified as type Va (noncalcified, n = 15) and type Vb (calcified, n = 15) in accordance with the American Heart Association consensus. Using laser capture microdissection, fibrous cap and shoulder regions were excised from frozen sections. Gene expression of pro- [interleukin 1 (IL-1), IL-8 and monocyte chemoattractant protein 1 (MCP-1)] and anti-inflammatory (IL-10) factors, and bone formation (bone morphogenetic protein 6 and osteocalcin) mediators were quantitated by real time PCR. Protein levels were determined using Western blotting. RESULTS: Mean percent carotid stenosis and calcification area were 79 and 5% in Va-plaques (40% symptomatic) and 77 and 42% in Vb-plaques (20% symptomatic). Macrophages infiltrating the region of the fibrous cap and the shoulder were more numerous in non-calcified plaques compared with calcified plaques (p < 0.01]. mRNA expression of MCP-1 and IL-8, and protein levels of IL-8 were also greater in Va plaques compared to Vb plaques (p < 0.05). Protein levels and mRNA expression of osteocalcin were greater in Vb compared to Va plaques (p < 0.05). CONCLUSIONS: Fibrous cap inflammation is more likely to occur in noncalcified than in calcified plaques. These findings suggest that carotid atherosclerotic plaque calcification is a structural marker of plaque stability. PMID- 19136824 TI - The dynamics of QT dispersion. PMID- 19136825 TI - Early reduction of QT dispersion after primary percutaneous intervention in ST segment elevation acute myocardial infarction. Mechanisms and clinical implications. AB - OBJECTIVES: To determine, in patients with ST-segment elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PPCI), the mechanisms and clinical implications of the acute changes in QT dispersion (QTd). METHODS: In this prospective study we included 216 patients admitted with a STEMI of <12 h of evolution. All were treated with PPCI. QTd was measured prior to PPCI and within 1 h after. RESULTS: The ratio of QTd reduction after PPCI (QTd-R) - defined as [(QTd before PPCI - QTd after PPCI)/QTd before PPCI] x100 - was significantly correlated with the percentage of ST-segment elevation resolution (ST-R; p < 0.001). To determine the significance of the different values of QTd-R, we further subdivided our population into 3 groups according to the tertiles of QTd-R (<10, 11-49, > or =50%). Patients with longer QTd-R had higher percentages of ST-R: 32 +/- 43 for QTd-R <10% vs. 60 +/- 21 for 11-49% vs. 71 +/- 12 for > or =50% (p < 0.05). By logistic regression, patients with QTd-R > or =50% had a reduction of 75% in the adjusted frequency of death or severe heart failure during hospitalization (95% CI 13-73%, p = 0.03). CONCLUSION: QTd-R after PPCI occurs early, is closely related to the restoration of reperfusion at the microvascular level and provides additional prognostic information. PMID- 19136826 TI - Variability of high-sensitivity C-reactive protein in chronic heart failure. PMID- 19136827 TI - Investigation of the effect of various fluoride compounds and preparations thereof on erosive tissue loss in enamel in vitro. AB - Metal fluoride preparations are promising in view of protection against dental erosion. The aim of this study was to screen the effect fluoride preparations containing various polyvalent metal cations had on erosion progression under increasingly severe erosive conditions in human enamel in vitro. Specimens were subjected to a cyclic de- and remineralisation procedure. Demineralisation (experiment E1 and E2a: 6 x 2 min/day, experiment E2b: 6 x 5 min/day) was performed with 0.05 M citric acid (pH 2.3). Fluoridation directly followed demineralisation and was reduced stepwise (E1: 6 x 2 min/day, E2a and E2b: 2 x 2 min/day). The test solutions were: ZnF(2 )(E1), AmF/NaF/CuSO(4) (E1), TiF(4)/AmF/Zn lactate (E1), TiF(4) (E1, E2a), TiF(4)/AmF (E2a), TiF(4)/AmF/NaF (E2a), TiF(4)/AmF/ZnF(2) (E1, E2a, E2b), AmF/NaF/SnCl(2) (2,800 mg/l Sn(2+); E1, E2a, E2b), AmF/NaF/SnCl(2)-1 (700 mg/l Sn(2+); E2a, E2b), all 1,500 ppm F(-), pH 4.5, and AmF/SnF(2) (positive control, pH 4.2, 250 ppm F(-); E1, E2a, E2b). In the negative control group, specimens were not fluoridated. Tissue loss was determined by profilometry after the last experimental day. Under mild conditions (E1), tin- and titanium-containing solutions were of marked efficacy and could nearly completely inhibit erosive tissue loss (reduction between 94.2 and 97.5%). With increasingly severe conditions (E2a, E2b), titanium lost its efficacy. Only the highly concentrated tin preparation (2,800 mg/l Sn(2+)) was able to reduce erosive tissue loss by 93.1%, even under severe conditions (E2b), with less frequent application of the experimental preparations. It should be considered for approaches for a symptomatic therapy of dental erosion. PMID- 19136828 TI - Effects of alpha-amylase and its inhibitors on acid production from cooked starch by oral streptococci. AB - This study evaluated acid production from cooked starch by Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinis and Streptococcus mitis, and the effects of alpha-amylase inhibitors (maltotriitol and acarbose) and xylitol on acid production. Streptococcal cell suspensions were anaerobically incubated with various carbohydrates that included cooked potato starch in the presence or absence of alpha-amylase. Subsequently, the fall in pH and the acid production rate at pH 7.0 were measured. In addition, the effects of adding alpha-amylase inhibitors and xylitol to the reaction mixture were evaluated. In the absence of alpha-amylase, both the fall in pH and the acid production rate from cooked starch were small. On the other hand, in the presence of alpha-amylase, the pH fell to 3.9-4.4 and the acid production rate was 0.61-0.92 micromol per optical density unit per min. These values were comparable to those for maltose. When using cooked starch, the fall in pH by S. sanguinis and S. mitis was similar to that by S. mutans and S. sobrinus. For all streptococci, alpha-amylase inhibitors caused a decrease in acid production from cooked starch, although xylitol only decreased acid production by S. mutans and S. sobrinus. These results suggest that cooked starch is potentially acidogenic in the presence of alpha-amylase, which occurs in the oral cavity. In terms of the acidogenic potential of cooked starch, S. sanguinis and S. mitis were comparable to S. mutans and S. sobrinus. Alpha-amylase inhibitors and xylitol might moderate this activity. PMID- 19136829 TI - Case-control study of early childhood caries in Australia. AB - The aim of this case-control study of 617 children was to investigate early childhood caries (ECC) risk indicators in a non-fluoridated region in Australia. ECC cases were recruited from childcare facilities, public hospitals and private specialist clinics to source children from different socioeconomic backgrounds. Non-ECC controls were recruited from the same childcare facilities. A multinomial logistic modelling approach was used for statistical analysis. The results showed that a large percentage of children tested positive for Streptococcus mutans if their mothers also tested positive. A common risk indicator found in ECC children from childcare facilities and public hospitals was visible plaque (OR 4.1, 95% CI 1.0-15.9, and OR 8.7, 95% CI 2.3-32.9, respectively). Compared to ECC-free controls, the risk indicators specific to childcare cases were enamel hypoplasia (OR 4.2, 95% CI 1.0-18.3), difficulty in cleaning child's teeth (OR 6.6, 95% CI 2.2-19.8), presence of S. mutans (OR 4.8, 95% CI 0.7-32.6), sweetened drinks (OR 4.0, 95% CI 1.2-13.6) and maternal anxiety (OR 5.1, 95% CI 1.1-25.0). Risk indicators specific to public hospital cases were S. mutans presence in child (OR 7.7, 95% CI 1.3-44.6) or mother (OR 8.1, 95% CI 0.9-72.4), ethnicity (OR 5.6, 95% CI 1.4-22.1), and access of mother to pension or health care card (OR 20.5, 95% CI 3.5-119.9). By contrast, a history of chronic ear infections was found to be protective for ECC in childcare children (OR 0.28, 95% CI 0.09-0.82). The biological, socioeconomic and maternal risk indicators demonstrated in the present study can be employed in models of ECC that can be usefully applied for future longitudinal studies. PMID- 19136830 TI - Performance of a pen-type laser fluorescence device and conventional methods in detecting approximal caries lesions in primary teeth--in vivo study. AB - This in vivo study aimed to compare the performance of different methods of approximal caries detection in primary molars. Fifty children (aged 5-12 years) were selected, and 2 examiners evaluated 621 approximal surfaces of primary molars using: (a) visual inspection, (b) the radiographic method and (c) a pen type laser fluorescence device (LFpen). As reference standard method, the teeth were separated using orthodontic rubbers during 7 days, and the surfaces were evaluated by 2 examiners for the presence of white spots or cavitations. The area under the receiver-operating characteristics curve (A(z)) as well as sensitivity, specificity and accuracy (percentage of correct diagnosis) were calculated and compared with the McNemar test at both thresholds. The interexaminer reproducibility was calculated using the intraclass correlation coefficient (ICC absolute values) and the kappa test (dichotomizing for both thresholds). The ICC value of the reference standard procedure was 0.94. At white-spot threshold, no methods tested presented good performance (sensitivity: visual 0.20-0.21; radiographic 0.16-0.23; LFpen 0.16; specificity: visual 0.95; radiographic 0.99 1.00; LFpen 0.94-0.96). At cavitation threshold, both LFpen and radiographic methods demonstrated higher sensitivity (0.55-0.65 and 0.65-0.70, respectively) and A(z) (0.92 and 0.88-0.89, respectively) than visual inspection sensitivity (0.30) and A(z) (0.69-0.76). All methods presented high specificities (around 0.99) and similar ICCs, but the kappa value for LFpen at white-spot threshold was lower (0.44). In conclusion, both LFpen and radiographic methods present similar performance in detecting the presence of cavitations on approximal surfaces of primary molars. PMID- 19136831 TI - Cognitive impairment and dementia in elderly people living in rural Benin, west Africa. AB - BACKGROUND/AIMS: Dementia is increasing as a priority public health problem because of the ageing of the world population. Our goal was to estimate dementia and cognitive impairment prevalence in an elderly population of rural Benin. METHODS: In a door-to-door survey, elderly people aged 65 years and above were screened using the Community Screening Interview for Dementia and the Five-Word Test. RESULTS: The prevalence of cognitive impairment was 10.4% and that of dementia was 2.6%. Age, current depressive disorder and absence of the APOE epsilon2 allele were significantly associated with cognitive impairment. CONCLUSION: Prevalence of dementia and cognitive impairment appears to be lower in this study than in developed countries. PMID- 19136832 TI - Cutaneous in vivo metabolism of topical lidocaine formulation in human skin. AB - Little is known about the metabolising capacity of the human skin in relation to topically applied drugs and formulations. We chose lidocaine as a model compound since the metabolic pathways are well known from studies concerning hepatic metabolism following systemic drug administration. However, the enzymes involved are also expressed in the skin. Hence, the aim of the current study was to investigate the extent of the cutaneous in vivo metabolism of topically applied lidocaine in human volunteers. A dose of 5 mg/cm(2) of Xylocaine(R) (5% lidocaine) ointment was applied onto the buttock skin of the volunteers. After 2 h, residual formulation was removed, and two 4-mm punch biopsies were taken from each volunteer. The quantity of lidocaine extracted from the skin samples (epidermis + dermis) was 109 +/- 43 ng/mm(2) skin. One metabolite (monoethylglycine xylidide, MEGX) was detected in skin from 7 of the 9 volunteers. The quantity of MEGX formed, relative to the quantity of lidocaine in the skin, was not consistent and ranged from <0.8 to 12.8%. No other metabolites were detected. PMID- 19136833 TI - Effects of various vehicles on skin hydration in vivo. AB - The stratum corneum, the outermost layer of the skin, regulates the passive loss of water to the environment. Furthermore, it is well accepted that drug penetration is influenced by skin hydration, which may be manipulated by the application of moisturizing or oleaginous vehicles. Measurements of transepidermal water loss (TEWL), and of skin hydration using a corneometer, were used to assess the effect of different vehicles on stratum corneum barrier function in vivo in human volunteers. A microemulsion significantly increased skin hydration relative to a reference vehicle based on medium chain triglycerides; in contrast, Transcutol(R) lowered skin hydration. TEWL measurements confirmed these observations. PMID- 19136834 TI - Abnormalities in stratum corneum function in patients recovered from leprosy. AB - BACKGROUND AND OBJECTIVES: Leprosy involves both the skin and peripheral nervous system. Leprosy patients display an increased incidence of xerosis and altered sensory thresholds, which persist in previously active skin sites. We assessed here whether alterations in stratum corneum (SC) function persist in cured leprosy, and the relationship of epidermal functional abnormalities to each clinical subtype of leprosy. METHODS: A total of 43 cured leprosy subjects and 29 normal control subjects were enrolled in this study. Basal skin surface pH, SC hydration, permeability barrier function as well as barrier recovery rates were measured over previously involved skin sites with a skin physiology monitor. One way ANOVA and two-tailed Student's t test were used to determine the significance between 2 groups and 3 or more groups, respectively. RESULTS: Competent barrier function was observed in all subtypes of cured leprosy subjects. All cured leprosy subjects except those with the borderline tuberculoid type exhibited a significantly lower SC hydration in comparison with normal subjects. Skin surface pH was significantly elevated in all cured leprosy subjects in comparison with normal subjects. CONCLUSIONS: A varied spectrum of alterations in SC function remains in all subjects who have recovered from leprosy, but the spectrum of SC functional abnormalities varies with disease subtype. PMID- 19136835 TI - Omega-6/omega-3 essential fatty acids: biological effects. PMID- 19136836 TI - Depression, suicide and deficiencies of omega-3 essential fatty acids in modern diets. PMID- 19136837 TI - Serial structural MRI analysis and proton and 31PMR spectroscopy in the investigation of cerebral fatty acids in major depressive disorder, Huntington's disease, myalgic encephalomyelitis and in forensic schizophrenic patients. PMID- 19136838 TI - Omega-3 fatty acid docosahexaenoic acid is the precursor of neuroprotectin D1 in the nervous system. PMID- 19136839 TI - Docosahexaenoic acid and amyloid-beta peptide signaling in Alzheimer's disease. PMID- 19136840 TI - Administration of docosahexaenoic acid, uridine and choline increases levels of synaptic membranes and dendritic spines in rodent brain. PMID- 19136841 TI - An assessment of dietary docosahexaenoic acid requirements for brain accretion and turnover during early childhood. PMID- 19136842 TI - Variation in lipid-associated genes as they relate to risk of advanced age related macular degeneration. PMID- 19136866 TI - Attorney involvement, claim duration, and workers' compensation costs. PMID- 19136868 TI - Public health responses to the risks of climate variability and change in the United States. AB - OBJECTIVE: Discuss issues related to the capacity of the United States to effectively adapt to current and future climate change. METHODS: Review literature on public health adaptation measures to reduce the burden of climate sensitive health outcomes. RESULTS: Most health risks of concern with climate change already exist in the United States. Current interventions may need to be augmented or deployed in new regions to prevent additional climate change-related morbidity and mortality. Monitoring and surveillance systems will need to be modified to ensure programs remain effective under a changing climate. CONCLUSIONS: Explicit consideration of climate change is needed in the many programs and research activities within federal, state, and local agencies that are relevant to adaptation to ensure that they have maximum effectiveness in reducing future vulnerability to the projected health impacts of climate change. PMID- 19136869 TI - Temperature extremes and health: impacts of climate variability and change in the United States. AB - OBJECTIVE: We evaluated temperature-related morbidity and mortality for the 2007 U.S. national assessment on impacts of climate change and variability on human health. METHODS: We assessed literature published since the 2000 national assessment, evaluating epidemiologic studies, surveys, and studies projecting future impacts. RESULTS: Under current climate change projections, heat waves and hot weather are likely to increase in frequency, with the overall temperature distribution shifting away from the colder extremes. Vulnerable subgroups include communities in the northeastern and Midwestern U.S.; urban populations, the poor, the elderly, children, and those with impaired health or limited mobility. CONCLUSIONS: Temperature extremes and variability will remain important determinants of health in the United States under climate change. Research needs include estimating exposure to temperature extremes; studying nonfatal temperature-related illness; uniform criteria for reporting heat-related health outcomes; and improving effectiveness of urban heat island reduction and extreme weather response plans. PMID- 19136870 TI - Climate change, extreme weather events, and us health impacts: what can we say? AB - OBJECTIVE: Address how climate change impacts on a group of extreme weather events could affect US public health. METHODS: A literature review summarizes arguments for, and evidence of, a climate change signal in select extreme weather event categories, projections for future events, and potential trends in adaptive capacity and vulnerability in the United States. RESULTS: Western US wildfires already exhibit a climate change signal. The variability within hurricane and extreme precipitation/flood data complicates identifying a similar climate change signal. CONCLUSIONS: Health impacts of extreme events are not equally distributed and are very sensitive to a subset of exceptional extreme events. Cumulative uncertainty in forecasting climate change driven characteristics of extreme events and adaptation prevents confidently projecting the future health impacts from hurricanes, wildfires, and extreme precipitation/floods in the United States attributable to climate change. PMID- 19136871 TI - Identifying vulnerable subpopulations for climate change health effects in the United States. AB - Climate change can be expected to have differential effects on different subpopulations. Biological sensitivity, socioeconomic factors, and geography may each contribute to heightened risk for climate-sensitive health outcomes, which include heat stress, air pollution health effects, extreme weather event health effects, water-, food-, and vector-borne illnesses. Particularly vulnerable subpopulations include children, pregnant women, older adults, impoverished populations, people with chronic conditions and mobility and cognitive constraints, outdoor workers, and those in coastal and low-lying riverine zones. For public health planning, it is critical to identify populations that may experience synergistic effects of multiple risk factors for health problems, both related to climate change and to other temporal trends, with specific geographic factors that convey climate-related risks. PMID- 19136872 TI - Multidomain subjective response to respirator use during simulated work. AB - OBJECTIVE: : Evaluate subjective tolerance to respirator use outside of traditional industrial settings by users including persons with mild respiratory impairment. METHODS: : The response to respirator use (half face mask dual cartridge and N95) was measured during eight types of work activities as well as in an exercise laboratory setting. The 43 research subjects included persons with mild respiratory impairments. Multiple domains of subjective response were evaluated. RESULTS: : Mixed model regression analyses assessing the effect of respirator type and task type showed: 1) most tolerated respirator use well; 2) half face mask respirators typically had greater adverse impact than N95 types; 3) multiple subjective outcomes, rather than only comfort/breathing impact, should be measured; and 4) rated subjective impact during work activities is less than in exercise laboratory settings. CONCLUSIONS: : The results suggest that respirator use may be feasible on a widespread basis if necessary in the face of epidemic or terror concerns. PMID- 19136873 TI - Independent and combined influence of physical activity and perceived stress on the metabolic syndrome in male law enforcement officers. AB - OBJECTIVE: : To examine the independent and combined interrelationships among physical activity, perceived stress, and the metabolic syndrome in law enforcement officers (LEOs). METHODS: : Perceived stress, self-reported physical activity, and metabolic syndrome risk factors were assessed in 386 white male LEOs. RESULTS: : Among the LEOs, 23.1% had the metabolic syndrome. The metabolic syndrome was not significantly associated with perceived stress (r = 0.047) whereas physical activity was (r = -0.225, P < 0.0001). The odds ratios (95% CI) for possessing the metabolic syndrome in the low and moderate physical activity groups compared to the high physical activity group were 3.13 (95% CI = 1.56 to 6.26) and 2.30 (95% CI = 1.29 to 4.09), respectively. CONCLUSIONS: : Regardless of stress level, physical inactivity is an important risk factor in the metabolic syndrome among this unique occupational group. PMID- 19136874 TI - A return-on-investment analysis of the health promotion program at the University of Minnesota. AB - OBJECTIVE: To determine the return-on-investment, if any, for the health promotion program adopted by the University of Minnesota in 2006. METHODS: Regression analysis was used to determine the cost-savings in annual health care expenditures associated with three components of the program: a risk assessment, a risk management program, and a disease management (DM) program. Differences-in differences equations with random effects were used to deal with selection bias. RESULTS: The analysis suggests that the DM reduced spending by about $1375 per year for each participant. The risk assessment and risk management components had no effect on spending in this initial year. CONCLUSIONS: DM reduced health care spending at the University of Minnesota, but not enough to generate a positive return-on-investment. A number of factors may qualify this conclusion. PMID- 19136875 TI - Direct and indirect costs to employers of patients with systemic lupus erythematosus with and without nephritis. AB - OBJECTIVES: To estimate the medical- and productivity-related cost burden of systemic lupus erythematosus (SLE) and SLE with nephritis in an employee population. METHODS: Using administrative data, annual costs were calculated for SLE patients, a subset with nephritis, and a matched control group of patients without SLE. These costs were compared with the cost of other chronic conditions. RESULTS: Mean annual medical expenditures and short term disability costs for SLE patients were $12,238 and $1184 greater (2005 dollars), respectively, than those of controls. Mean medical expenditures for SLE/nephritis patients were $46,862 greater than for controls. When compared with other chronic health conditions faced by employees, SLE/nephritis was the most costly condition. CONCLUSIONS: SLE, particularly with nephritis, is associated with substantial costs. Therapies that can better manage SLE may provide opportunities for savings to employers. PMID- 19136876 TI - Screening tools to estimate mold burdens in homes. AB - OBJECTIVE: The objective of this study was to develop screening tools that could be used to estimate the mold burden in a home which would indicate whether more detailed testing might be useful. METHODS: Two possible screening methods were considered for mold analysis: use of vacuum cleaner bag dust rather than the standard protocol dust samples and reducing the number of molds needed to be quantified resulting in the creation of an alternative mold burden scale. RESULTS: Vacuum bag dust analysis placed the estimate of mold burden into the upper or lower half of the Environmental Relative Moldiness Index scale. Mold burdens estimated by only 12 species produced an index, the American Relative Moldiness Index, with a correlation of rho = 0.80 with the Environmental Relative Moldiness Index. CONCLUSIONS: Two screening tools were developed for estimating the mold burden in homes. PMID- 19136877 TI - Work experiences and tobacco use: findings from the gear up for health study. AB - OBJECTIVE: To examine the relationships between the work environment and amount smoked, intention to quit, and participation in a health promotion intervention. METHODS: Cross-sectional data were collected from unionized truck drivers and dockworkers (N = 542; response rate = 78%) employed in eight trucking terminals as part of a larger intervention study targeting tobacco use cessation and weight management. RESULTS: Prevalence of tobacco use was 40% (n = 216). Multivariable analyses of amount smoked and coworker norms encouraging cessation found significant associations with intention to quit and negative social consequences of tobacco use. Program participation was significantly associated with concern about job exposures, working the day shift, and intention to quit. CONCLUSIONS: Work experiences represented in the social contextual model may help explain how the work environment affects tobacco-use behaviors and interest in quitting, and maybe guide interventions among blue-collar workers. PMID- 19136878 TI - Inflammatory responses and oxidative stress from metal fume exposure in automobile welders. AB - OBJECTIVES: : Welding operations expose to metal fume. We investigated the inflammatory responses and oxidative stress from metal fume exposure in an automobile plant. METHODS: : We recruited 258 automobile workers and measured the urine zinc, copper, and nickel to determine the exposure level, and examined the white blood cells, and IL-6 as inflammatory responses to the metal fume exposure. We also examined the relationship between glutathione (GSH) and metals exposure. RESULTS: : There were significant association between urine metals levels and welding hours. Zinc was significantly associated with blood white cells, interleukin-6, and GSH. Copper was significantly associated with GSH, but nickel was significantly inversely associated with GSH. CONCLUSIONS: : Automobile welders appear to have significant metals exposure. White blood cells and IL-6 might be involved in inflammatory process of zinc fume exposure with zinc and copper increasing GSH, but nickel depleting it. PMID- 19136879 TI - Predicting sickness absence-are extended health check-ups of any value? Comparisons of three individual risk models. AB - OBJECTIVE: To predict sickness absence by three health check-up models. METHODS: A study group of 821 participants from the public sector in Sweden where three health check-up models were compared 1) the limited variable model including smoking, body mass index, blood pressure, and cholesterol, 2) the several variable model including smoking, waist-hip ratio, blood pressure, relation between low density lipoproteins and high density lipoproteins, serum triglycerides, and fitness test, and 3) Self-rated health measured by one single question. Sickness absence data during 1 year was delivered from the employer. RESULTS: The three models served their purpose to predict sickness absence. CONCLUSIONS: The self-rated health-model with one single question has as good quality in predestination as more complicated models. This may have an implication for cost-effective procedures in occupational health services. PMID- 19136880 TI - Healthy workforce/healthy economy: the role of health, productivity, and disability management in addressing the nation's health care crisis: why an emphasis on the health of the workforce is vital to the health of the economy. PMID- 19136881 TI - Why are occupational medicine and industrial hygiene complementary professions and what makes them the key members of the occupational health team? PMID- 19136883 TI - Cellular loss after allogenic hepatocyte transplantation. AB - Hepatocyte transplantation (HCTx) has been considered as an alternative to orthotopic liver transplantation for the treatment of acute and chronic liver failure and metabolic liver diseases. One of the factors that mainly limit its application is the significant cellular loss after HCTx. Although liver has been considered as an immunologically privileged organ, transplanted hepatocytes can only survive for 7 to 10 days without immunosuppression. In fact, both innate and adaptive immune system are implicated in the destruction to allografts, which is a particular type of rejection. This article has reviewed the progress in the mechanisms of cellular loss after allogenic HCTx. PMID- 19136884 TI - Pretransplant genetic testing of live kidney donors at risk for autosomal dominant polycystic kidney disease. PMID- 19136885 TI - Dimerization of soluble HLA-G by IgG-Fc fragment augments ILT2-mediated inhibition of T-cell alloresponse. AB - BACKGROUND: Human leukocyte antigen (HLA)-G, a nonclassical HLA class I molecule, induces a wide range of tolerogenic immunological effects by means of interaction with its inhibitory receptors. However, recent studies show that HLA-G dimer formation is essential to bind to its receptors and exhibit its effects. METHODS: In this study, a soluble divalent HLA-G/IgG molecule (sHLA-G dimer) was constructed. Its inhibitory effect on T-cell alloresponse was studied with mixed lymphocyte reaction in vitro, which was set up by mixing inactivated T1 cells with HLA-mismatched peripheral blood lymphocytes in the presence or absence of the sHLA-G dimer. RESULTS: The results show that sHLA-G dimer inhibits T-cell alloresponse by reducing proliferation of both CD4+ and CD8+ T cells and suppressing generation of alloreactive cytotoxic T lymphocytes at nanomole concentration. The inhibition of the sHLA-G dimer is observed to be more effective than that of sHLA-G monomer. Our results also indicate that sHLA-G dimer up-regulates inhibitory receptor ILT2 on alloreactive CD8+ T cells, which contributes to the significant inhibition on T-cell alloresponse. CONCLUSION: The sHLA-G dimer formed by IgG-Fc fragment shows more inhibitory effects on alloreactive T cells, which may have implications for allotransplantation. PMID- 19136887 TI - Significance of serum bile acids in small bowel allograft rejection in pigs. AB - OBJECTIVES: To examine changes in individual bile acids in serum in a pig model of allogenic small bowel transplantation. METHODS: Seventeen pigs were divided into two groups: group 1 (n=10), controls; group 2 (n=7), allotransplantation, nonimmunosuppressed recipients. Both groups received a segmental intestine. Intestinal specimens for histologic studies were obtained at the end of cold flushing (T0) and on postoperative day 8 (T1). Total and individual bile acid concentrations were measured in serum samples taken at T0 and T1. RESULTS: All animals survived until the end of the study. In group 1, intestinal histology revealed no significant changes between T0 and T1 specimens. In contrast, in group 2, graft histology revealed moderate to severe rejection at T1 in all specimens. In contrast to group 1, serum levels of total bile acids increased significantly in group 2 at T1 compared with baseline (P=0.001). The increase was due to secondary bile acids which were significantly higher at T1 compared with baseline (P=0.0003). In contrast, secondary bile acids increased but not significantly at T1 in group 1 compared with baseline (P=0.056). CONCLUSION: Our study suggests that total and secondary serum bile acids may be a useful diagnostic tool for the diagnosis of acute intestinal rejection. PMID- 19136888 TI - Cryopreserved agarose-encapsulated islets as bioartificial pancreas: a feasibility study. AB - BACKGROUND: A bioartificial pancreas in which islets of Langerhans (islets) are encapsulated within a semipermeable membrane, such as agarose, has been proposed for treating type I diabetic mellitus. However, the long-term storage for providing a convenient and easily accessible supply still remains an issue. We investigated cryopreservation as a potential method of long-term storage for agarose-encapsulated islets (Mic-islets). METHODS: The morphology, insulin secretion, and histochemical staining of cryopreserved Mic-islets were analyzed. Streptozotocin-induced diabetic mice were transplanted intraperitoneally with 1000, 2000, and 3000 Mic-islets after cryopreservation in KYO-1 vitrification solution. Blood glucose levels were measured and immunohistochemical analyses were performed at 41 days posttransplantation. RESULTS: Transplanted cryopreserved Mic-islets restored normoglycemia in diabetic mice. The mean (+/ SD) normoglycemic periods were 32.0+/-13.2 days and 46.3+/-13.3 days for recipients of 1000 (n=5) and 2000 (n=4) cryopreserved Mic-islets, respectively, whereas the mean normoglycemic period was 53.2+/-16.7 days for recipients of 1000 noncryopreserved Mic-islets (n=7). These data indicate that cryopreserved Mic islets transplanted as a bioartificial pancreas successfully controlled blood glucose levels for extended periods. CONCLUSION: Cryopreserved agarose encapsulated islets could successfully control the blood glucose level for a long period as a bioartificial pancreas. PMID- 19136886 TI - Differential roles of direct and indirect allorecognition pathways in the rejection of skin and corneal transplants. AB - BACKGROUND: It is generally accepted that all transplants are not rejected in the same fashion. However, the extrinsic and intrinsic factors that control the recognition and rejection of a particular allograft by the host are not well characterized. METHODS: We compared the mechanisms underlying the response with donor antigens by T cells activated after transplantation of fully allogeneic skin and corneal grafts in mice. RESULTS: In corneal-transplanted mice, the CD4+ T-cell response was exclusively mediated by T cells recognizing minor antigens in an indirect fashion and producing low levels of interleukin-2. In contrast, skin grafts elicited both direct and indirect CD4+ T-cell responses primarily directed to major histocompatibility complex antigens and characterized by high interleukin-2 levels. Although CD8+ T-cells producing gamma interferon were activated directly in both skin- and corneal-grafted mice, only CD8+ T cells from skin-transplanted mice mounted a cytotoxic response. Next, we investigated whether failure of corneal transplants to induce a CD4+ direct alloresponse is due to their poor immunogenicity or due to the site of placement (eye). We observed that corneas transplanted under the skin and splenocytes transplanted in the eye were both capable of inducing direct CD4+ T-cell alloreactivity. CONCLUSIONS: This shows that failure of orthotopic corneal allotransplants to elicit a CD4+ T-cell direct alloresponse is associated with the combination of two factors, their low immunogenicity and the immune-privileged properties of the eye. PMID- 19136890 TI - Galectin-3-mediated xenoactivation of human monocytes. AB - BACKGROUND: Delayed xenograft rejection (DXR) remains a roadblock to successful xenotransplantation. A feature of DXR is early recruitment of monocytes to the xenograft. Naive human monocytes can recognize and adhere to unstimulated porcine aortic endothelial cells (PAEC) more than human aortic endothelial cells, partly due to endothelial expression of the xenoantigen galactose-alpha(1,3)galactose beta(1,4)GlcNAc-R (alpha-gal). Previous work from our laboratory has implicated galectin-3 as a candidate molecule on monocytes involved in initial recognition and adhesion of human monocytes to PAEC. METHODS: Flow cytometry was used to analyze monocyte activation and galectin-3 accumulation in PAEC. Reactive oxygen intermediate production was analyzed using dihydrorhodamine measured in a fluorescence plate reader. Western blotting was performed to determine galectin-3 secretion and expression by human monocytes. Immunofluorescence staining for the tight junction protein zona occludens-1 was used as a measure of PAEC monolayer integrity. RESULTS: We demonstrate that galectin-3 can be secreted from monocyte intracellular stores on contact with alpha-gal. Soluble galectin-3 binds PAEC partly by expression of alpha-gal. Binding is reduced on endothelium derived from alpha-gal knockout animals, but not completely. Competing terminal sugars expressed on human aortic endothelial cells such as sialic acid, may block galectin-3 binding. Furthermore, soluble galectin-3 activates monocytes in an autocrine/paracrine manner. Blocking galectin-3 reduces the activation of human monocytes. Finally, the inhibition of galectin-3 reduces monocyte-mediated endothelial injury on co-culture with PAEC. CONCLUSION: Galectin-3 plays a role in human monocyte activation and adhesion in the presence of PAEC, which may contribute to DXR. Additional transgenic strategies targeting galectin-3 ligands on porcine endothelium may be required to achieve optimal xenograft survival. PMID- 19136889 TI - HLA-E/human beta2-microglobulin transgenic pigs: protection against xenogeneic human anti-pig natural killer cell cytotoxicity. AB - BACKGROUND: Natural killer (NK) cells participate in pig-to-primate xenograft rejection both by antibody-dependent and -independent mechanisms. A majority of human NK cells express the inhibitory receptor CD94/NKG2A, which binds specifically to human leukocyte antigen (HLA)-E, a trimeric complex consisting of the HLA-E heavy chain, beta2-microglobulin (beta2m), and a peptide derived from the leader sequence of some major histocompatibility complex class I molecules. METHODS: To use this mechanism for protection of pig tissues against human NK cell-mediated cytotoxicity, we generated transgenic pigs by pronuclear microinjection of genomic fragments of HLA-E with an HLA-B7 signal sequence and of human beta2-microglobulin (hubeta2m) into zygotes. RESULTS: Three transgenic founder pigs were generated. Northern blot analysis of RNA from peripheral blood mononuclear cells revealed the presence of the expected transcript sizes for both transgenes in two of the three founders. The founder with the highest expression and his offspring were characterized in detail. Fluorescence-activated cell sorting (FACS) and Western blot analyses demonstrated consistent expression of HLA-E and hubeta2m in peripheral blood mononuclear cells. Immunohistochemistry revealed the presence of HLA-E and hubeta2m on endothelial cells of many organs, including heart and kidney. In vitro studies showed that lymphoblasts and endothelial cells derived from HLA-E/hubeta2m transgenic pigs are effectively protected against human NK cell-mediated cytotoxicity, depending on the level of CD94/NKG2A expression on the NK cells. Further, HLA-E/hubeta2m expression on porcine endothelial cells inhibited the secretion of interferon (IFN)-gamma by co cultured human NK cells. CONCLUSIONS: This novel approach against cell-mediated xenogeneic responses has important implications for the generation of multitransgenic pigs as organ donors for clinical xenotransplantation. PMID- 19136891 TI - The additional detrimental effects of cold preservation on transplantation associated injury in kidneys from living and brain-dead donor rats. AB - BACKGROUND: Brain death and cold preservation are major alloantigen-independent risk factors for transplantation outcome. The present study was conducted to assess the influence of these factors on transplantation-associated injury independently or in combination. METHODS: Brain death was induced in F344 rats. Renal grafts were harvested after 6 hr and either directly transplanted in unilateral nephrectomized Lewis recipient or subjected to 24 hr of cold preservation in University of Wisconsin solution before implantation. Allografts obtained from living donor rats were also subjected to cold preservation or not. DNA damage was assessed before implantation by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling staining. Ten days after transplantation, renal histology was performed according to Banff '97 classification. The expressions of cytokines and adhesion molecules were analyzed by quantitative polymerase chain reaction. RESULTS: Cold preservation significantly increased the number of terminal deoxynucleotide transferase mediated dUTP nick-end labeling positive cells in renal allografts. Ten days after transplantation, histology revealed a higher degree of tubulitis and vasculitis scores when the grafts were subjected to cold storage. Vasculitis was aggravated when the graft was obtained from brain death (BD) donors. BD, but not cold preservation alone, was associated with papillary necrosis. This was more frequently observed after cold preservation. Immunohistology showed an increase in MHC class II+ cells after cold preservation. The combination of BD and cold preservation revealed a higher degree of VEGF and IL-10 expression. CONCLUSIONS: Our Study emphasizes that cold ischemia time should be limited when renal allografts from brain-dead donors are transplanted. PMID- 19136892 TI - Protein kinase C inhibitor, AEB-071, acts complementarily with cyclosporine to prevent islet rejection in rats. AB - BACKGROUND: AEB-071 (AEB) is a specific inhibitor of protein kinase C, which prevents T-lymphocyte activation. The present study investigated the effect of AEB on rat islet allotransplantation alone or in combination with CTLA4-Ig, mycophenolate mofetil, or cyclosporine A (CsA). METHODS: A rodent allogeneic islet transplant model (Lewis to Wistar Furth) was used to investigate the efficacy of AEB as an immunosuppressive agent. Furthermore, the Lewis rat was used to screen for any AEB associated toxicities on glucose homeostasis in vivo. RESULTS: AEB alone (30 mg/kg per os [p.o.] two times per day [bid]) delayed rejection to a median survival time of 22 days (vs. 7 days in control vehicle treated animals, P<0.05). When combined with CsA (5 mg/kg p.o. bid), AEB prolonged survival from 12 (CsA alone) to over 100 days in 80% of animals (P<0.05). No delay in allograft rejection (above that resulting from AEB alone) was observed when AEB was combined with a sub-therapeutic dose of CTLA4-Ig or mycophenolate mofetil, nor low dose of CsA. The frequency of allospecific interferon-gamma-secreting splenocytes, assessed ex vivo by enzyme-linked immunosorbent spot (ELISPOT) assay, was lower in AEB-treated recipients compared with controls (P<0.05). AEB treatment did not alter the intraperitoneal glucose tolerance, the glucose-dependent insulin release, or the insulin content of the native pancreas. CONCLUSIONS: These data suggest that AEB is an appropriate immunosuppressive agent for islet transplantation, as it can prolong islet graft survival alone or in combination with CsA, without toxicity on glucose metabolism. PMID- 19136893 TI - Intraperitoneal insulin infusion versus islet transplantation: comparative study in patients with type 1 diabetes. AB - OBJECTIVE: The purpose of this article was to compare two strategies for insulin delivery in brittle type 1 diabetic patients: intraperitoneal insulin infusion (IPII) through an implantable pump and intraportal islet transplantation (IIT). METHODS: Thirteen consecutive patients (6 islet after kidney and 7 islet transplantation alone), treated with IIT according to the Edmonton protocol, were compared with 17 patients treated with IPII in the same center in a nonrandomized study. Both groups of patients were assessed for clinical profile, metabolic results, and adverse events during the 3-year period after implantation. RESULTS: Sex ratio, mean age, body mass index, diabetes duration, daily insulin need (DIN), blood creatinine, lipid and HbA1c levels, and frequency of diabetic complications did not differ significantly between the two groups before inclusion. The comparison of metabolic results 3, 6, and 12 months after IIT or IPII showed that while mean HbA1c significantly decreased over time in both groups, mean DIN, glycemia, and number of hypoglycemias less than 3.3 mmol/L per patient per week (Hypo) only significantly decreased in the IIT group versus baseline. At 12 months, mean DIN, HbA1c, and Hypo were significantly lower in the IIT versus IPII group. After 24 and 36 months, mean DIN, HbA1c, and Hypo remained significantly lower in the IIT group versus baseline, and mean HbA1c and DIN versus IPII. Adverse events were, however, fourfold more frequent with IIT versus IPII, though their numbers decreased over time. CONCLUSION: These results suggest that metabolic results improve with both methods, but were significantly better with IIT versus IPII, though with more frequent side effects. PMID- 19136894 TI - Fibrous intimal thickening at implantation adversely affects long-term kidney allograft function. AB - BACKGROUND: Kidneys from marginal and older donors are increasingly used to respond to the increasing demand for kidney transplants. This study evaluated the predictive value of intimal hyperplasia, as a marker of vasculopathy, in the renal allograft at the time of transplantation (transplantation) on the subsequent graft function (7 years). METHODS: The intima/media ratio of the arterial walls (I/M) was morphometrically determined by the sectorial elliptic method, in 51 implantation biopsies. Two study groups were determined. Group 1, with I/M less than or equal to 0.47, was considered as the group with minimal vascular damage at transplantation. Group 2, with I/M more than 0.47, was considered as having vasculopathy at transplantation. RESULTS: During the first 15 months, the estimated glomerular filtration rate improved in group 1 from 53+/ 17 to 61+/-17 mL/min/1.73 m2, whereas it decreased from 51+/-21 to 46+/-14 mL/min/1.73 m2 in group 2. From 1 year posttransplantation, the estimated glomerular filtration rate (eGFR) was significantly higher in group 1 at all time points (6 month evaluation). The difference in graft function between the two groups (mean, 11 mL/min/1.73 m2) remained unchanged between 1 and 7 years posttransplantation. Among several clinical parameters investigated, blood pressure of the recipient significantly modulates the impact of preexisting vasculopathy on graft function. CONCLUSION: Our data provide evidence that donor related vasculopathy, at the time of transplantation, has a persistent significant impact on the subsequent graft function. This effect becomes only apparent at 1 year posttransplantation and is increased in recipients with inadequately controlled blood pressure. PMID- 19136896 TI - Pediatric liver transplantation for metabolic liver disease: experience at King's College Hospital. AB - AIMS: The aims of this article were to report a single-center experience of pediatric liver transplantation for liver-based metabolic disorders and to compare the outcome of cirrhotic versus noncirrhotic metabolic liver disease. METHODS: The medical records of 96 patients younger than 18 years undergoing transplantation for liver-based metabolic disorders from 1989 to 2005 were reviewed. RESULTS: Hundred twelve transplants were performed in 96 patients at a median age of 59.7 months (range, 0-208 months). The cumulative 1-, 5-, and 10 year graft and patient survival rates were 83%, 77%, and 62% and 91%, 86%, and 82%, respectively. Acute liver failure at first presentation (hazard ratio [HR] 3.0; 95% confidence interval [CI] 1.1-8.1), age less than 1 year at time of transplantation (HR 4.6; 95% CI 1.7-12.4) and hospitalization (HR 3.2; 95% CI 1.1 9.3) were significant predictors of worse patient survival. For noncirrhotic disorders, the long-term patient (100% vs. 100%, 90% vs. 100%, and 90% vs. 75%, P=0.87) and graft survivals (93% vs. 100%, 70% vs. 100%, and 70 vs. 75%, P=0.12) at 1, 5, and 7 years for auxiliary versus orthotopic transplantation were not significantly different. CONCLUSIONS: Long-term patient survival after transplantation for metabolic disorders is excellent for both cirrhotic and noncirrhotic metabolic disorders. For noncirrhotic metabolic disorders, auxiliary transplantation has similar patient and graft survival compared with orthotopic transplantation, but further research is recommended. PMID- 19136895 TI - The effect of the JAK inhibitor CP-690,550 on peripheral immune parameters in stable kidney allograft patients. AB - INTRODUCTION: CP-690,550 inhibits Janus kinase 3 (JAK3) which mediates signal transduction of receptors of the common gamma-chain cytokines. These cytokines play key roles in lymphocyte function and homeostasis. As part of a phase 1 trial, we evaluated the effect of CP-690,550 on immune parameters. MATERIAL: Stable kidney transplant recipients (n=8) receiving mycophenolate mofetil and prednisolone were treated with CP-690,550, 30 mg twice daily orally for 29 days. Blood samples were collected on days 1 (before first dose), 15, 29 (end of treatment), and 57. RESULTS: Two patients experienced minor infections (one urinary tract infection and one mild respiratory tract infection). Leukocyte counts remained stable, whereas a mean decrease in hemoglobulin of 8% was measured (P=0.01). CP-690,550 treatment for 29 days resulted in statistically significant changes in the number of circulating CD19+ B cells (P=0.05), CD3- CD16+ CD56+ natural killer-cells (P<0.01), and CD4+ CD25bright+ T cells (P=0.05; one-way analysis of variance). After CP-690,550 treatment on day 15 the number of B cells increased by a mean of 100%, (P=0.04), whereas those of natural killer cells and CD4+ CD25bright+ T cells decreased by 65% (P=0.001) and 38% (P=0.03, t test), respectively, from pretreatment baseline. However, the regulatory capacities of the residual CD4+ CD25bright+ T cells remained unchanged pre- and posttreatment. In addition, in the presence of CP-690,550, the interferon-[gamma] production capacity of peripheral blood mononuclear cells was reduced by 39% (median) compared with predose baseline (P=0.01). CONCLUSIONS: These findings demonstrate the role of JAK3 in the homeostasis and function of select lymphocyte subpopulations. JAK3 inhibition may provide a novel mechanism for the modulation of allogeneic responses in patients after transplantation. PMID- 19136897 TI - Donor-specific antibodies against HLA, MICA, and GSTT1 in patients with allograft rejection and C4d deposition in renal biopsies. AB - BACKGROUND: Production of antibodies against donor-specific antigens is one of the central mechanisms of allograft rejection. This antibody-mediated rejection (AMR) is evidenced by the presence of circulating donor-specific antibodies and deposition of complement component C4d on renal endothelium. Although anti-human leukocyte antigen (HLA) antibodies account for a high proportion of AMR, in many cases anti-HLA antibodies cannot be demonstrated. In liver transplant, antibodies against glutathione-S-transferase T1 (GSTT1) expressed on the graft may induce an antibody response leading to a severe graft dysfunction. In addition, presence of antibodies against major-histocompatibility-complex class I chain-related gene A (MICA) has been associated with a poor graft survival in kidney transplantation. METHODS: Pre- and posttransplantation sera from 19 patients fulfilling the criteria for AMR including C4d deposition in renal biopsies were included. Donor specific antibodies against HLA-I and -II and MICA were studied using Luminex. Anti-GSTT1 antibodies were analyzed by indirect immunofluorescence and by an ELISA method. A control group of 39 patients with graft dysfunction negative for C4d was also included. RESULTS: At the time of the biopsy, 4 (21%) patients had only anti-HLA class I antibodies; 3 (15.8%) had anti-GSTT1, 2 (10.5%) had anti HLA-class II, and 2 (10.5%) had anti-MICA; four patients had combination of antibodies: HLA-I + MICA (n=1), HLA-I + GSTT1 (n=2), and GSTT1+MICA (n=1). No antibodies were found in 4 (21%) patients. In total, 6 (31.6%) C4d+ patients had anti-GSTT1 antibodies, whereas, among the 39 C4d-negative patients, only 3 (7.7%) had anti-GSTT1 antibodies (P=0.027). CONCLUSION: Besides anti-HLA antibodies, donor-specific antibodies against MICA and GSTT1 antigens could be responsible for the occurrence of antibody-mediated kidney graft rejection. PMID- 19136899 TI - Adoptive immune transfer of hepatitis B virus specific immunity from immunized living liver donors to liver recipients. AB - BACKGROUND: Liver transplantation is often the ultimate option of therapy for chronically hepatitis B virus (HBV) infected patients. Prevention of reinfection is therapy intensive and cost-effective. Adoptive transfer of HBV-specific immunity with the liver from an immune living liver donor (LLD) could be a new approach to prevent reinfection. METHODS: Forty-six potential LLDs were vaccinated against HBV. Humoral (antibodies to hepatitis B virus surface antigen [anti-HBs]-titer) and cellular (IFN-gamma-ELISpot and proliferation-assay) immune responses were examined in donors after immunization and in recipients before and after transplantation. RESULTS: Anti-HBs-titers of up to 50,000 IU/L were detected in LLDs. Fourteen recipients received livers from these donors. We detected humoral immunity in one HBV-naive recipient and in one chronically HBV infected recipient after transplantation. A transfer of cellular immunity (SI>3) was seen in three recipients. These patients received livers from donors with high anti-HBs-titers of more than 9000 IU/L. Cellular immunity was also detected in the corresponding donors (SI >3 and spots >22). CONCLUSIONS: Our study demonstrates that HBV-specific humoral and cellular immunity can be transferred by liver transplantation after vaccination of the donors. The transfer of B-cell and T-cell immunity correlates with the magnitude of immune responses in the donor. PMID- 19136898 TI - Kidney transplant half-life (t[1/2]) after rapid discontinuation of prednisone. AB - Protocols incorporating rapid discontinuation of prednisone (RDP) after kidney transplantation have been associated with good short-term results. However, concern remains that RDP will be associated with decreased long-term graft survival rates. We compared kidney transplant half-life (t1/2) for recipients treated with antibody induction, calcineurin inhibitor, antimetabolite, and RDP versus historical controls treated with antibody induction, calcineurin inhibitor, antimetabolite, and maintenance prednisone. For both living and deceased donor recipients, we found no difference between groups. We also found no differences in rate of graft loss to acute rejection or to tubular atrophy and interstitial fibrosis. Our study suggests that long-term graft outcome is not decreased when using RDP protocols versus chronic maintenance prednisone. PMID- 19136900 TI - Cerebrovascular events in renal transplant recipients. AB - BACKGROUND: The incidence of stroke and risk factors for different subtypes of cerebrovascular (CBV) events in renal transplant recipients have not been examined in any large prospective controlled trial. METHODS: The Assessment of Lescol in Renal Transplantation was a randomized, double-blind, placebo controlled study of the effect of fluvastatin (40-80 mg) daily on cardiovascular, and renal outcomes in renal transplant recipients. Patients initially randomized to fluvastatin or placebo in the 5 to 6 year trial was offered open-label fluvastatin in a 2-year extension to the original study. We investigated the incidence of stroke and risk factors for ischemic and hemorrhagic CBV events in 2102 renal graft recipients participating in the Assessment of Lescol in Renal Transplantation core and extension trial with a mean follow-up of 6.7 years. RESULTS: The incidence and type of CBV events did not differ between the lipid lowering arm and the placebo arm. A total of 184 (8.8%, 95% confidence interval 4.6-12.9) of 2102 patients experienced a CBV event during follow-up, corresponding to an incidence of 1.3% CBV event per year. The mortality for patients experiencing a hemorrhagic stroke was 48% (13 of 27), whereas the mortality for ischemic strokes was 6.0% (8 of 133). Diabetes mellitus, previous CBV event, age, and serum creatinine were independent risk factors for cerebral ischemic events. The risk of a hemorrhagic cerebral event was increased by diabetes mellitus, polycystic kidney disease, left ventricular hypertrophy, and systolic blood pressure. INTERPRETATION: Risk factors for CBV events in renal transplant recipients differ according to subtype. PMID- 19136901 TI - Calcitriol deficiency and 1-year mortality in cardiac transplant recipients. AB - BACKGROUND: Administration of the vitamin D hormone calcitriol improves survival in solid-organ transplanted experimental animals. We investigated whether lower serum calcitriol concentrations are associated with increased 1-year mortality in cardiac transplant recipients. METHODS: We prospectively recruited 171 patients who underwent cardiac transplantation at out institution between May 2004 and April 2006. We assessed calciotropic hormones, inflammation markers, and renal function preoperatively and on postoperative days 6 (t1) and 21 (t2). RESULTS: Serum creatinine and C-reactive protein increased, whereas calcitriol decreased significantly after transplantation (P<0.001). As determined by multivariable Cox regression analysis, the calcitriol level at t2 was an independent predictor of 1 year mortality. One-year mortality was 3.7 per 100 person-years in the tertile with the highest calcitriol concentrations at t2 (> 18 pg/mL), 13.2 per 100 person-years in the intermediate tertile (11-18 pg/mL), and 32.1 per 100 person years in the tertile with the lowest calcitriol concentrations at t2 (< 11 pg/mL) (P<0.001). 25-Hydroxyvitamin D deficiency (serum concentrations below 10 ng/mL), renal insufficiency (serum creatinine > or = 1.6 mg/dL), and high serum concentrations of the inflammation markers C-reactive peptide and tumor necrosis factor-alpha were predictors of a serum calcitriol concentration below 11 pg/mL (P=0.037-0.001). CONCLUSIONS: Low postoperative calcitriol concentrations are independently associated with high 1-year mortality in cardiac transplant recipients. A causal relationship has yet to be proven by intervention trials using active vitamin D. PMID- 19136902 TI - Alemtuzumab induction and antibody-mediated kidney rejection after simultaneous pancreas-kidney transplantation. AB - BACKGROUND: The best induction agent for simultaneous pancreas-kidney transplantation (SPKT) remains the subject of debate. Alemtuzumab is effective in preventing acute cellular rejection (ACR) in SPK recipients and has been used to prevent antibody-mediated rejection (AMR) in sensitized kidney transplant candidates. METHODS: A retrospective cohort study was performed including 136 SPK recipients receiving maintenance immunosuppression with tacrolimus, mycophenolic acid prodrugs, and prednisone. Two groups were compared: those who received induction with alemtuzumab (n=97) and those induced with basiliximab (n=39). RESULTS: Kidney ACR was more frequent in SPKT induced with basiliximab (2-year 12.8% vs. 3.1%, P=0.04), but the incidence of AMR was similar (2-year 18% with basiliximab vs. 13.8% with alemtuzumab, P=NS). Kidney rejection was associated with clinical pancreas rejection in 70% of cases, without differences between the groups. Postrejection kidney graft survival was similar in both groups (2-year basiliximab/alemtuzumab 94.7%/91.2%), but death-censored kidney graft survival was lower with alemtuzumab (100%/91.2%, P=0.056). In the basiliximab group, the predominant cause of kidney loss was death-with-function, whereas in the alemtuzumab group AMR accounted for all losses. Pancreas graft survival was similar in both groups, yet more pancreas losses due to acute rejection occurred in alemtuzumab-treated patients (4 vs. 1). CONCLUSIONS: Kidney AMR is more common than ACR in SPKT recipients treated with alemtuzumab, tacrolimus, mycophenolic acid, and steroids. ACR is better prevented by alemtuzumab than basiliximab, but no relevant difference is found in prevention of AMR. Despite the high incidence of AMR, survival rates are excellent in both groups. PMID- 19136904 TI - FoxP3 positive T cells in graft biopsies from living donor kidney transplants after donor-specific transfusions. AB - Donor-specific transfusions (DST) induce allograft tolerance in animals. Evidence is growing that FoxP3+ regulatory T cells are associated with tolerance in humans. Forty-four biopsies from 69 living donor kidney transplant recipients (LDT) after DST, 53 biopsies from 69 matched deceased donor transplant recipients (DDT), obtained for graft dysfunction, and 12 biopsies from LDT without DST were retrospectively analyzed. FoxP3 positivity was more frequent in LDT/DST than in DDT biopsies (67% vs. 44%, P=0.02). Considering only biopsies with acute rejection, FoxP3 positivity was observed in 92% (11/12) after LDT/DST, but only in 50% (6/12) after DDT (P=0.03). The number of FoxP3+ T cells per total infiltrating cells in rejection biopsies was higher (P<0.05) from LDT/DST (4.1%) than from DDT or LDT (2.6%) without DST (2.5%). Six-year graft survival was better in patients with LDT/DST than with DDT (87.5% vs. 79.7%, P=0.04). The present investigation demonstrates an association between DST and FoxP3+ T cells. The effect of DST on regulatory T cells deserves further analysis in transplantation. PMID- 19136903 TI - DNA testing for live kidney donors at risk for autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is characterized by age dependent growth of kidney cysts with end-stage renal disease developing in approximately 50% of affected individuals. Living donors from ADPKD families are at risk for developing ADPKD and may be excluded from renal donation if the diagnosis cannot be conclusively ruled out. Radiographic imaging may be adequate to screen for kidney cysts in most at-risk donors but may fail to identify affected individuals younger than 40 years or older individuals from families with mild disease. In this article, we report a strategy that incorporates genetic testing in the evaluation of live kidney donors at risk for ADPKD whose disease status cannot be established with certainty on the basis of imaging studies alone. We show that DNA diagnostics can be used to enhance safe donation for certain living donor candidates at risk for ADPKD. PMID- 19136905 TI - Proteomic analysis of HCV cirrhosis and HCV-induced HCC: identifying biomarkers for monitoring HCV-cirrhotic patients awaiting liver transplantation. AB - BACKGROUND: Progression from chronic hepatitis C virus (HCV) infection to cirrhosis and hepatocellular carcinoma (HCC) results in protein changes in the peripheral blood. We evaluated global protein expression in plasma samples of HCV cirrhotic and HCV-cirrhotic-HCC patients. PATIENTS AND METHODS: Plasma samples from 25 HCV-cirrhotic-HCC and 10 HCV-cirrhotic patients were quantitatively evaluated for protein expression. Tryptic peptides were analyzed using Thermo linear ion-trap mass spectrometer (LTQ) coupled with a Surveyor HPLC system (Thermo). SEQUEST and X!Tandem database search algorithms were used for peptide sequence identification. Protein relative quantification was performed using the area under the curve from the select ion chromatogram. A significant fold change between groups was based on controlling the false discovery rate (FDR) at less than 5%. RESULTS: We identified and quantified 2320 proteins from the analysis of the different protein pattern between HCV-cirrhosis and HCV-HCC samples. Gene ontology terms classified the more important biologic process related to these proteins as signal transduction, regulation of transcription DNA-dependent, protein amino acid phosphorylation, cell adhesion, transport, and immune response. Seven proteins showed significant expression changes with a FDR less than 5% between cirrhosis and tumor groups. Moreover, 18 proteins showed significant expression changes (FDR <5%) when plasma samples from HCV-cirrhosis were compared with early HCV-HCC. CONCLUSIONS: Differential protein expression was observed between samples from HCV patients with cirrhosis with and without HCC. Also, differences were observed between early and advanced HCV-HCC samples. This study provides important information for discovery of potential biomarkers for early HCC diagnosis in HCV cirrhotic patients. PMID- 19136906 TI - Exenatide use in islet transplantation: words of caution. PMID- 19136909 TI - Non-surgical tissue biopsy among patients with advanced pancreatic cancer: effect on survival. AB - OBJECTIVE: The aim was to determine the survival of patients with advanced, unresectable pancreatic cancer in relation to whether they underwent nonsurgical biopsy of their primary tumor. METHODS: A total of 1481 patients with distant stage pancreatic cancer diagnosed between 1992 and 2001 who underwent radiation treatment but not cancer-directed surgery were analyzed. The design is a retrospective cohort study from the Surveillance, Epidemiology, and End Results program of the US National Cancer Institute. Survival curves were created using Kaplan-Meier method and compared via log-rank test. RESULTS: Of 1481 patients (median age, 66 years) included in our analysis, 1406 (95%) underwent nonsurgical biopsy (95%) and 75 (5%) did not. There was no statistically significant difference in overall median survival according to receipt of nonsurgical biopsy (Kaplan-Meier curve, log-rank test = 0.09). A subgroup analysis of patients younger than 65 years who did not undergo biopsy revealed a hazard ratio of 1.76 (95% confidence interval, 1.14-2.72); that is, there was a 76% higher hazard for death among younger patients who did not undergo biopsy compared with those who did (P = 0.011). CONCLUSION: Nonsurgical biopsy did not seem to negatively impact survival among patients with advanced pancreatic cancer. PMID- 19136908 TI - Proteomic analyses of pancreatic cyst fluids. AB - OBJECTIVES: There are currently no diagnostic indicators that are consistently reliable, obtainable, and conclusive for diagnosing and risk-stratifying pancreatic cysts. Proteomic analyses were performed to explore pancreatic cyst fluids to yield effective diagnostic biomarkers. METHODS: We have prospectively recruited 20 research participants and prepared their pancreatic cyst fluids specifically for proteomic analyses. Proteomic approaches applied were as follows: (1) matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry peptidomics with LC/MS/MS (HPLC-tandem mass spectrometry) protein identification; (2) 2-dimensional gel electrophoresis; (3) GeLC/MS/MS (tryptic digestion of proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by LC/MS/MS). RESULTS: Sequencing of more than 350 free peptides showed that exopeptidase activities rendered peptidomics of cyst fluids unreliable; protein nicking by proteases in the cyst fluids produced hundreds of protein spots from the major proteins, making 2-dimensional gel proteomics unmanageable; GeLC/MS/MS revealed a panel of potential biomarker proteins that correlated with carcinoembryonic antigen (CEA). CONCLUSIONS: Two homologs of amylase, solubilized molecules of 4 mucins, 4 solubilized CEA-related cell adhesion molecules (CEACAMs), and 4 S100 homologs may be candidate biomarkers to facilitate future pancreatic cyst diagnosis and risk stratification. This approach required less than 40 microL of cyst fluid per sample, offering the possibility to analyze cysts smaller than 1 cm in diameter. PMID- 19136910 TI - Defatted bovine colostrum-supplemented diet around weaning improves exocrine pancreatic secretion by means of volume, digestive enzymes, and antibacterial activity. AB - OBJECTIVES: Exocrine pancreatic secretion contributes to limit pathogenic bacteria-associated diarrhea. Bovine colostrum, used in the treatment of diarrhea, reduces symptoms originating from gut pathogenic bacteria overgrowth. We hypothesized that bovine colostrum may stimulate the exocrine pancreatic secretion. METHODS: Eighteen piglets fitted with 2 permanent catheters (for pancreatic juice collection and reintroduction) were allocated to 1 of the following 2 dietary treatments for 5 days: a control diet or a diet supplemented with defatted bovine colostrum. Pancreatic juice was collected daily, and digestive enzyme activities and antibacterial activity were determined. RESULTS: The prandial pancreatic juice outflow, the basal and prandial lipase output, and the basal secretion of the antibacterial activity were, respectively, 60% (P = 0.08), 154% (P = 0.08), 92% (P = 0.06), and 72% (P < 0.05) higher in piglets fed a diet supplemented with defatted bovine colostrum. CONCLUSIONS: With defatted bovine colostrum, the increased antibacterial activity secretion against Escherichia coli may limit pathogenic bacteria overgrowth of the gut and reduce diarrheal episodes. The role of secretin in the increased pancreatic juice flow and lipase secretion was considered. PMID- 19136911 TI - Chemical composition of essential oils of Thymus and Mentha species and their antifungal activities. AB - The potential antifungal effects of Thymus vulgaris L., Thymus tosevii L., Mentha spicata L., and Mentha piperita L. (Labiatae) essential oils and their components against 17 micromycetal food poisoning, plant, animal and human pathogens are presented. The essential oils were obtained by hydrodestillation of dried plant material. Their composition was determined by GC-MS. Identification of individual constituents was made by comparison with analytical standards, and by computer matching mass spectral data with those of the Wiley/NBS Library of Mass Spectra. MIC's and MFC's of the oils and their components were determined by dilution assays. Thymol (48.9%) and p-cymene (19.0%) were the main components of T. vulgaris, while carvacrol (12.8%), a-terpinyl acetate (12.3%), cis-myrtanol (11.2%) and thymol (10.4%) were dominant in T. tosevii. Both Thymus species showed very strong antifungal activities. In M. piperita oil menthol (37.4%), menthyl acetate (17.4%) and menthone (12.7%) were the main components, whereas those of M. spicata oil were carvone (69.5%) and menthone (21.9%). Mentha sp. showed strong antifungal activities, however lower than Thymus sp. The commercial fungicide, bifonazole, used as a control, had much lower antifungal activity than the oils and components investigated. It is concluded that essential oils of Thymus and Mentha species possess great antifungal potential and could be used as natural preservatives and fungicides. PMID- 19136912 TI - Synthesis of 2,3-dioxo-5-(substituted)-arylpyrroles and their 2-oxo-5-aryl-3 hydrazone pyrrolidine derivatives. AB - Some novel 2,3-dioxo-5-(substituted)-arylpyrroles have been synthesized. Among these, pyrrolidine compound 1b was converted to 2,3-dioxo-5-aryl pyrrolidine 2b. Finally a set of hydrazone derivatives was obtained from the reaction of 2b with various hydrazine salts. The structures of all the new synthesized compounds were confirmed by elemental analyses, IR and 1H-NMR spectra. PMID- 19136913 TI - Molecular docking studies and anti-tyrosinase activity of Thai mango seed kernel extract. AB - The alcoholic extract from seed kernels of Thai mango (Mangifera indica L. cv. 'Fahlun') (Anacardiaceae) and its major phenolic principle (pentagalloylglucopyranose) exhibited potent, dose-dependent inhibitory effects on tyrosinase with respect to L-DOPA. Molecular docking studies revealed that the binding orientations of the phenolic principles were in the tyrosinase binding pocket and their orientations were located in the hydrophobic binding pocket surrounding the binuclear copper active site. The results indicated a possible mechanism for their anti-tyrosinase activity which may involve an ability to chelate the copper atoms which are required for the catalytic activity of tyrosinase. PMID- 19136914 TI - Isolation of betulinic acid, its methyl ester and guaiane sesquiterpenoids with protein tyrosine phosphatase 1B inhibitory activity from the roots of Saussurea lappa C.B.Clarke. AB - Activity-guided fractionation of a MeOH extract of the roots of Saussurea lappa C.B.Clarke (Compositae), using an in vitro protein tyrosine phosphatase 1B (PTP1B) inhibition assay, led to the isolation of four active constituents: betulinic acid (1), betulinic acid methyl ester (2), mokko lactone (3) and dehydrocostuslactone (4), along with nine inactive compounds. Our findings indicate that betulinic acid (1) and its methyl ester 2, as well as the two guaiane sesquiterpenoids 3 and 4 are potential lead moieties for the development of new PTP1B inhibitors. PMID- 19136915 TI - The main chemical composition and in vitro antifungal activity of the essential oils of Ocimum basilicum Linn. var. pilosum (Willd.) Benth. AB - The essential oils of the aerial parts of Ocimum basilicum Linn.var. pilosum (Willd.) Benth., an endemic medicinal plant growing in China, was obtained by hydrodistillation and analysed by GC-MS. Fifteen compounds, representing 74.19% of the total oil were identified. The main components were as follows: linalool (29.68%), (Z)-cinnamic acid methyl ester (21.49%), cyclohexene (4.41%), alpha- cadinol (3.99%), 2,4-diisopropenyl-1-methyl-1-vinylcyclohexane (2.27%), 3,5 pyridine-dicarboxylic acid, 2,6-dimethyl-diethyl ester (2.01%), beta-cubebene (1.97%), guaia-1(10),11-diene (1.58%), cadinene (1.41%) (E)-cinnamic acid methyl ester (1.36%) and beta-guaiene (1.30%). The essential oils showed significant antifungal activity against some plant pathogenic fungi. PMID- 19136916 TI - Screening of agelasine D and analogs for inhibitory activity against pathogenic protozoa; identification of hits for visceral leishmaniasis and Chagas disease. AB - There is an urgent need for novel and improved drugs against several tropical diseases caused by protozoa. The marine sponge (Agelas sp.) metabolite agelasine D, as well as other agelasine analogs and related structures were screened for inhibitory activity against Plasmodium falciparum, Leishmania infantum, Trypanosoma brucei and T. cruzi, as well as for toxicity against MRC-5 fibroblast cells. Many compounds displayed high general toxicity towards both the protozoa and MRC-5 cells. However, two compounds exhibited more selective inhibitory activity against L. infantum (IC(50) <0.5 microg/mL) while two others displayed IC(50) <1 microg/mL against T. cruzi in combination with relatively low toxicity against MRC-5 cells. According to criteria set up by the WHO Special Programme for Research & Training in Tropical Diseases (TDR), these compounds could be classified as hits for leishmaniasis and for Chagas disease, respectively. Identification of the hits as well as other SAR data from this initial screening will be valuable for design of more potent and selective potential drugs against these neglected tropical diseases. PMID- 19136917 TI - Antioxidant phenolic substances of Turkish red wines from different wine regions. AB - In this study, five biologically phenolic antioxidant Turkish red wines from different regions of Turkey were determined using HPLC with PDA detection. The antioxidant capacities (AC) of the investigated wines are also determined and the relationship between the phenol content and antioxidant capacity is discussed. The results show that Kalecik Karasi, Merlot and Cabernet Sauvignon AC values ranged between 15.8-18.7 mmol/L, 15.8-17.6 mmol/L and 18.1-22.6 mmol/L, respectively. Generally, Cabernet Sauvignon wines not only had higher levels of phenolic antioxidants, such as catechin, epicatechin and gallic acid, but also higher antioxidant capacities compared to Kalecik Karasi and Merlot wines. When the results were compared on the basis of geographical area, Aegean red wines were found to contain generally higher levels of biologically important phenolics and thus to have more antioxidant capacity compared to the wines of the other regions studied. PMID- 19136919 TI - Routine, single session, indirect laser for proliferative diabetic retinopathy. AB - AIMS: To evaluate the visual and morphological outcomes of routine, single session, indirect panretinal photocoagulation (PRP) for proliferative diabetic retinopathy, and to examine adverse events related to indirect laser within the first 8 weeks of treatment. METHODS: The case notes of 107 diabetics with proliferative retinopathy undergoing indirect PRP were reviewed retrospectively, and compared with the UK National Diabetic Retinopathy Laser Treatment Audit. Patients who had received PRP previously were excluded. Follow-up data were collected as close as possible to 9 months following the initial laser treatment. RESULTS: Fifteen patients (14.0%) returned with adverse events within the first 8 weeks of indirect PRP. There were two tractional retinal detachments and two new vitreous haemorrhages. One patient with underlying neurotrophic keratopathy developed a persistent epithelial defect. One developed non-progressive macular drag with a one-line drop in Snellen acuity. Three experienced new or exacerbations of preexisting clinically significant macular oedema, which resolved spontaneously. There were two transient choroidal effusions, two transient anterior uveitis, and two transient visual deteriorations without macular oedema clinically. There were no significant differences in the rates of new tractional retinal detachment, vitreous haemorrhage or rubeosis, but our vitrectomy rate was higher (7.5 vs 1.5%, P=0.02, chi2), largely due to our policy of performing vitrectomy early. CONCLUSIONS: Visual and morphological outcomes of routine, single session, indirect PRP for proliferative diabetic retinopathy were not inferior to the outpatient-based national audit, and the incidence of significant PRP-induced adverse events was low. PMID- 19136918 TI - Methodologies for the extraction of phenolic compounds from environmental samples: new approaches. AB - Phenolic derivatives are among the most important contaminants present in the environment. These compounds are used in several industrial processes to manufacture chemicals such as pesticides, explosives, drugs and dyes. They also are used in the bleaching process of paper manufacturing. Apart from these sources, phenolic compounds have substantial applications in agriculture as herbicides, insecticides and fungicides. However, phenolic compounds are not only generated by human activity, but they are also formed naturally, e.g., during the decomposition of leaves or wood. As a result of these applications, they are found in soils and sediments and this often leads to wastewater and ground water contamination. Owing to their high toxicity and persistence in the environment, both, the US Environmental Protection Agency (EPA) and the European Union have included some of them in their lists of priority pollutants. Current standard methods of phenolic compounds analysis in water samples are based on liquid liquid extraction (LLE) while Soxhlet extraction is the most used technique for isolating phenols from solid matrices. However, these techniques require extensive cleanup procedures that are time-intensive and involve expensive and hazardous organic solvents, which are undesirable for health and disposal reasons. In the last years, the use of news methodologies such as solid-phase extraction (SPE) and solid-phase microextraction (SPME) have increased for the extraction of phenolic compounds from liquid samples. In the case of solid samples, microwave assisted extraction (MAE) is demonstrated to be an efficient technique for the extraction of these compounds. In this work we review the developed methods in the extraction and determination of phenolic derivatives in different types of environmental matrices such as water, sediments and soils. Moreover, we present the new approach in the use of micellar media coupled with SPME process for the extraction of phenolic compounds. The advantages of micellar media over conventional extractants are reduction of organic solvent, low cost, easy handling and shorter time procedures. PMID- 19136920 TI - A prospective ultrasound biomicroscopy evaluation of changes in anterior segment morphology following laser iridotomy in European eyes. AB - PURPOSE: The aim of this study is to quantify changes in anterior segment morphology by the use of ultrasound biomicroscopy (UBM) after Nd:YAG laser iridotomy in primary angle closure (PAC) and PAC glaucoma (PACG) in European eyes. METHODS: A total of 35 eyes of 28 consecutive patients presenting with PAC or PACG were examined by UBM at presentation, and 1 week after Nd:YAG laser peripheral iridotomy (LPI). Mean age of patients was 63.5+/-14.4 (SD) years. Seventeen patients were females (60%). The trabecular-iris angle (TIA) was measured in the superior, nasal, inferior, and temporal quadrants. Baseline measurements were made under light and dark conditions. MAIN OUTCOME MEASURE: UBM measurements of the trabecular-iris angle (TIA) and the angle opening distance (AOD). DESIGN: Consecutive observational case series. RESULTS: All measurements were made in four quadrants. Before LPI, mean superior TIA was 3.59+/-4.5 degrees (mean+/-SD), nasal TIA was 6.37+/-4.9 degrees , inferior TIA was 9.33+/-7.6 degrees , and temporal TIA was 8.65+/-7.3 degrees in light conditions. After LPI, these values increased, respectively, to 12.58+/-6.9 degrees (P<0.05), 15.40+/ 6.8 degrees (P<0.05), 16.37+/-7.4 degrees (P<0.05), and 15.95+/-11.3 degrees (P<0.05), showing a significant widening of the angle in all four quadrants. Superior AOD increased from 0.060+/-0.07 to 0.107+/-0.07 mm (P=0.09). No serious LPI-related complications were encountered. CONCLUSIONS: Dimensions of the anterior chamber angle can be significantly influenced by Nd:YAG laser iridotomy in narrow angle European eyes. UBM examination is a viable tool for the quantitative evaluation of the anterior chamber angle before and after laser iridotomy. PMID- 19136921 TI - Risk of Creutzfeldt-Jakob disease transmission by ocular surgery and tissue transplantation. AB - Creutzfeldt-Jakob disease (CJD) is a rare, fatal neurodegenerative disease that occurs in sporadic, genetic, variant, and iatrogenic forms. The transformation of normal prion protein (PrP(C)) to the abnormal form (PrP(Sc)) is a key step in the pathogenesis of CJD and leads to the accumulation of amyloid and spongiform changes in the brain. The presence of PrP(Sc) in tissue is a surrogate marker for CJD infectivity. Sporadic CJD, whose cause is unknown, is by far the most frequent form with 1-2 cases per million population occurring every year-the genetic forms of CJD are rather rarer. The majority of variant CJD cases have occurred in the United Kingdom, where there have been four reports of transmission of vCJD by blood transfusion. The great majority of iatrogenic transmissions of CJD have resulted from the use of pituitary-derived hormones or dura mater with only a very few cases attributable to neurosurgical instruments or corneal transplants. In the absence of a validated test for CJD infectivity in eye donors, the application of appropriate donor selection criteria and the use of single-use instruments in eye banks are currently the most effective means of reducing the risk of CJD transmission. Onward transmission by reusable ophthalmic surgical instruments has not been reported, but the risk cannot be excluded. Use of appropriate cleaning and disinfection protocols and the ability to identify and quarantine instruments that may have been used on an infected patient are important safeguards. PMID- 19136922 TI - Ocular immune privilege. AB - It has been over 60 years since the phrase immune privilege was used by Sir Peter Medawar to describe the lack of an immune response against allografts placed into the ocular microenvironment. Since then, we have come to understand that the mechanisms of ocular immune privilege include unique anatomical features of a blood barrier and a lack of direct lymphatic drainage. Also, we know that the ocular microenvironment is rich with immunosuppressive molecules that influence the activity of immune cells. Moreover, the placement of foreign antigen into the ocular microenvironment can induce a systemic form of tolerance to the foreign antigen called anterior chamber-associated immune deviation (ACAID). Many soluble immunomodulators are found in aqueous humour, and are a mixture of growth factors, cytokines, neuropeptides, and soluble receptors. This is a continuously growing list. The mechanisms of ocular immune privilege induce apoptosis, promote the production of anti-inflammatory cytokines, and mediate the activation of antigen-specific regulatory immunity. These mechanisms of immune privilege also attempt to impose themselves upon immunity within the uveitic eye. The adaptation of several anatomical and biochemical mechanisms to establish an immune privileged microenvironment within the eye makes the eye immunologically unique. It is a tissue site where we may learn how immunity is regulated in inflammation and at rest. Success in translating the lessons of ocular immune privilege to other tissues has the potential to drastically change the therapy and clinical outcomes of autoimmune diseases and allograft survival. PMID- 19136923 TI - Antibody-based immunosuppressive agents for corneal transplantation. AB - The progress in antibody engineering over the last 20 years has created the tools for the development of novel antibody-based drugs and constructs, such as small antibody fragments, suitable for topical administration. In rheumatology, oncology, transplantation medicine and ophthalmology, therapeutic antibody constructs, and antibody fragments have been responsible for the clinical progress seen over the last decade. Although antibody-based therapies have become a well-established immunosuppressive option in solid organ transplantation, there are only very few reports with regard to corneal transplantation. The following review explains some of the important aspects of engineered antibody-based therapeutic agents and summarises the current use of such immunosuppressive therapies in transplantation medicine and corneal transplantation. PMID- 19136924 TI - Ipsilateral rotational autokeratoplasty: a review. AB - Corneal opacity is a major cause of monocular blindness and, after cataract, is also a leading cause of blindness worldwide. Keratoplasty techniques for the treatment of corneal opacities include deep anterior lamellar allokeratoplasty, penetrating allokeratoplasty, penetrating bilateral autokeratoplasty, and ipsilateral rotational autokeratoplasty (IRA). This review describes the indications, technique, and outcomes of IRA. IRA is only indicated for patients with a localised opacity leaving a minimum diameter of 4-5 mm of uninvolved clear cornea. For these few patients in whom the procedure is practicable, the surgery can be planned by manipulating digital images to estimate the trephine size and location and/or by the use of formulas. IRA may not provide either as good spectacle acuity or as good quality of vision as penetrating keratoplasty because of higher astigmatism and a reduced corneal pupillary clear zone, but these disadvantages are often outweighed when the risk of allograft rejection is high, as in paediatric patients and those with vascularised corneas. The main benefits of IRA are the retention of host endothelium, thereby eliminating both the risk of endothelial rejection and the prolonged attrition of endothelial cell numbers that occurs following penetrating keratoplasty, and the reduced requirement for postoperative steroid therapy with its associated complications. PMID- 19136925 TI - Is measurement of adult height useful in screening for primary angle closure? AB - BACKGROUND: Adult height is independently related to ocular dimensions and shorter people have shorter globes and shallower anterior chambers. We investigated the relationship between adult height and angle dimensions to explore measuring height as a possible screening test for angle closure. METHODS: A population-based, cross-sectional survey of adult Chinese living in the Tanjong Pagar district, Singapore was conducted. Anterior chamber depth, gonioscopic angle width and height were assessed using standardized protocols. RESULTS: Data were available for 996 persons. Shorter people had shallower anterior chamber depth (2.35 mm for persons <144 cm vs 2.72 mm for persons >170 cm, P=0.008) and smaller gonioscopic angles (22 degrees for <144 cm vs 30 degrees for >170 cm, P=0.079). After controlling for age and gender, adult height was significantly related to anterior chamber depth (P=0.008) but not significantly related to gonioscopic angle width (P=0.079). Female sex and age > or = 50 years used together correctly identified 45/66 (68.2%) individuals with an occludable angle (sensitivity 68.2%, specificity 61.3%). Fewer people, 41/66 (62.1%), were correctly identified when height <160 cm was added to female sex and age 50 > or = years (sensitivity was 62.1% and specificity was 64.7%). CONCLUSIONS: Shorter adult height is significantly associated with a shallower anterior chamber depth, but the addition of height contributed little to demographics (age and gender) as the preliminary screening criteria to identify individuals at risk of an occludable angle. PMID- 19136926 TI - Hypomethylation of long interspersed nuclear element-1 in hepatocellular carcinomas. AB - Recent studies have revealed the epigenetic alterations are involved in hepatocarcinogenesis. However, the function of long interspersed nuclear element 1 hypomethylation in hepatocellular carcinomas, and relationship among other clinicopathologic features, and genetic and epigenetic alterations, including CpG island hypermethylation, have not been studied. We determined long interspersed nuclear element-1 methylation, a marker of global methylation, in 57 tumor and nonneoplastic samples, including 24 from high-risk and 33 from low-risk countries. We compared methylation levels of long interspersed nuclear element-1 with eight CpG islands including p16, cyclooxygenase-2, T-type calcium channel, and estrogen receptor genes, and MINT31, MINT1, MINT2, and MINT27, as well as CpG island methylator phenotype and p53 gene mutation. Most hepatocellular carcinomas samples (88%) showed hypomethylation of long interspersed nuclear element-1, with a mean level of global methylation of 58+/-14 compared to 77+/-6 in nonneoplastic hepatic tissue (P<0.001). Levels of long interspersed nuclear element-1 hypomethylation differed depending on geographic location (P=0.02), status of hepatitis (P=0.01), hypermethylation of p16, estrogen receptor and MINT2 (P=0.01, 0.002, and 0.045, respectively), CpG island methylator phenotype-positive status (P=0.006), and p53 gene mutation (P=0.04). In conclusion, environmental factors such as geographic location and hepatitis status contribute to hepatocarcinogenesis through global hypomethylation. In hepatocellular carcinomas, hypermethylation of CpG islands, and CpG island methylator phenotype status seems to correlate with levels of long interspersed nuclear element-1 hypomethylation. PMID- 19136927 TI - The role of Movat pentachrome stain and immunoglobulin G4 immunostaining in the diagnosis of autoimmune pancreatitis. AB - Autoimmune pancreatitis is highly responsive to steroid therapy, but because it mimics pancreatic cancer, it often precipitates unnecessary surgery. Adequate diagnostic tests are needed to permit appropriate medical therapy. Lymphocytic and obliterative phlebitis are reported in the majority of cases, as are elevated IgG4-positive plasma cells, indicating their high sensitivity. Their specificities, especially when used in conjunction, however, remain largely unknown. Movat pentachrome vascular and IgG4 immunohistochemical stains were performed on a total of 15 autoimmune pancreatitis cases (11 pancreatic resections and 4 biopsies), 39 usual-type alcoholic or idiopathic chronic pancreatitis cases, 35 pancreatic ductal adenocarcinoma-associated chronic pancreatitis cases, and 29 normal pancreata. Marked and diffuse lymphocytic and obliterative venulitis were detected in all 15 cases of autoimmune pancreatitis on Movat staining (100% sensitivity). Only a single carcinoma-associated chronic pancreatitis case among all of the controls showed diffuse benign venulitis that was nonobliterative (99% specificity). Nine of 13, 9 autoimmune pancreatitis cases showed marked IgG4 immunopositivity at >or=10 positive plasma cells per x 400 field (69% sensitivity). No increased IgG4 plasma cells were found in any of 103 controls (100% specificity). In combination, all of the autoimmune pancreatitis cases had at least one (13/13) and most had both markers (9/13), whereas none of the controls had both markers. Overall, these combined stains show very promising diagnostic utility and should be considered in combination with clinical and serologic analyses in the evaluation of chronic pancreatitis suspicious for malignancy. Future validating studies on preoperative biopsies with outcome data following steroid therapy will be essential. PMID- 19136928 TI - Overexpression of HMGA2 relates to reduction of the let-7 and its relationship to clinicopathological features in pituitary adenomas. AB - High-mobility group A2 is highly expressed during embryogenesis and in various benign and malignant tumors. Recent studies report that high-mobility group A2 is negatively regulated by the let-7 microRNAs (miRNAs) family in vitro. The development of pituitary adenomas in high-mobility group A2 transgenic mice showed that high-mobility group A2 may be involved in pituitary tumorigenesis. However, no studies have investigated the clinical significance of high-mobility group A2 and its relationship to the let-7 miRNA family in human pituitary adenomas. Using immunohistochemistry, we analyzed high-mobility group A2 expression with respect to various clinicopathologic factors in 98 pituitary adenomas. Overexpression of high-mobility group A2 was observed in 39% (38/98) of pituitary adenomas compared with normal adenohypophysial tissue and was frequently found in adenomas including prolactin (PRL), adrenocorticotrophic hormone, or follicle-stimulating hormone/luteinizing hormone and in null cell adenomas, but relatively rare in growth hormone (GH) and mixed GH/PRL adenomas. High-mobility group A2 expression was significantly associated with tumor invasion (P<0.05) and was significantly higher in grade IV than in grades I, II, and III adenomas (P<0.05). High levels of high-mobility group A2 expression were more frequently observed in macroadenomas than in microadenomas (P<0.05). High levels of high-mobility group A2 expression also significantly correlated with the proliferation marker Ki-67 (P<0.0001). Real-time quantitative RT-PCR analysis was carried out to evaluate the expression of let-7 in 55 pituitary adenomas. Subsequently, decreased expression of let-7 was confirmed in 23 of 55 (42%) adenomas and was correlated with high-grade tumors (P<0.05). An inverse correlation between let-7 and high-mobility group A2 expression was evident (R= 0.33, P<0.05). These findings support a causal link between let-7 and high mobility group A2 whereby loss of let-7 expression induces high-mobility group A2 upregulation that represents an important mechanism in pituitary tumorigenesis and progression. PMID- 19136929 TI - Hypermutation in mantle cell lymphoma does not indicate a clinical or biological subentity. AB - Mantle cell lymphoma is a prime example of a well-defined entity based on morphology, phenotype, genetics and also clinical features. Although most patients have an adverse clinical course, some have a better survival than others. The most consistently reported adverse prognostic parameter is a high mitotic rate. Recently, it has been shown that hypermutation in the immunoglobulin heavy-chain gene occurs in a subset of mantle cell lymphomas. It is, however, unclear whether the mutational status is stable over time within a given case, whether hypermutation might be influenced by therapy and how it is related to other relevant biological features of mantle cell lymphoma. In this study, we analyzed 23 typical mantle cell lymphoma cases with respect to mutational status and compared the results with clinicopathological and genetic data to determine whether the presence of mutation indicates a subentity with clinical or pathological relevance. We found somatic hypermutation in 26% of our cases and, interestingly, one case showed ongoing somatic hypermutation. In tumor cells of both mutated and unmutated cases, we found a preferential usage of V(H)3 21 (23%) and V(H)4-34 (19%). No significant correlations were found between mutation status and the other morphological and genetic features analyzed. In conclusion, our results provide additional evidence that mutation status in mantle cell lymphoma is better interpreted as a feature within the spectrum of disease that seems to have little clinical or pathological relevance. PMID- 19136930 TI - Tumor deposits are encountered in advanced colorectal cancer and other adenocarcinomas: an expanded classification with implications for colorectal cancer staging system including a unifying concept of in-transit metastases. AB - The seventh edition of the TNM Classification of Malignant Tumors is due to be published soon. In the current version dating back to 2002, tumor deposits, which are metastatic lesions commonly encountered in the routine histopathological examination of advanced colorectal cancer specimens, are classified according to their shape with different implications for staging. So distinguished, these lesions are considered either as metastatic lymph nodes (N category) or as vascular invasions (T category). We recently proposed a more comprehensive classification approach that also includes the M category. Relying on two of our independent recent studies, we aim here to provide suggestions for a novel classification of tumor deposits with diverse implications for TNM staging system of colorectal cancer. Furthermore, we show that tumor deposits are not limited to colorectal cancer, but they are common to different adenocarcinoma types. PMID- 19136931 TI - Recurrent numerical aberrations of JAK2 and deregulation of the JAK2-STAT cascade in lymphomas. AB - The Janus kinase 2 (JAK2)-signal transducers and activators of transcription (STAT) pathway plays an important role in hematological malignancies. Mutations and translocations of the JAK2 gene, mapped at 9p24, lead to constitutive activation of JAK2 and its downstream targets. The presence of JAK2 mutations in lymphomas has been addressed in larger cohorts, but there are little systemic data on numerical and structural JAK2 aberrations in lymphoid neoplasms. To study the molecular epidemiology of these aberrations and the consecutive activation of the JAK2-STAT pathway in lymphomas, we examined 527 cases, covering the most common entities, in a tissue microarray by fluorescent in situ hybridization with breakable JAK2 probes, and immunohistochemistry for phosphorylated JAK2 (pJAK2) and its preferred downstream pSTAT3 and pSTAT5. 9p24 gains were detected in 6/17 (35%) primary mediastinal B-cell lymphomas (PMBCLs), 25/77 (33%) Hodgkin's lymphomas (HLs), 3/16 (19%) angioimmunoblastic T-cell lymphomas (AILTs) and 1/5 ALK1(+) anaplastic large cell lymphomas (ALCLs); breaks were observed only in three cases. pJAK2 expression was most prevalent in PMBCL, peripheral T-cell lymphomas and HL. pSTAT3 predominated in ALCLs, HLs, AILTs, PMBCLs and peripheral T-cell lymphomas. pSTAT5 expression was detected frequently in follicular lymphomas, diffuse large B-cell lymphomas and AILTs. 9p24 gains correlated with increased proportions of tumor cells expressing pJAK2 (P=0.002) and pSTAT3 (P=0.001). In follicular lymphomas, concomitant expression of pJAK2 and pSTAT5 was linked to better prognosis, whereas expression of pSTAT3 in nongerminal center-like diffuse large B-cell lymphomas could identify a patient group with an inferior outcome. Our findings stress that despite the rarity of activating JAK2 mutations in lymphomas, JAK2 is recurrently targeted by numerical, and rarely by structural, genetic aberrations in distinct lymphoma subtypes and that JAK2-STAT pathway may play a role in lymphomagenesis. PMID- 19136932 TI - IGF2BP3 (IMP3) expression is a marker of unfavorable prognosis in ovarian carcinoma of clear cell subtype. AB - Clear cell carcinoma is an uncommon subtype of ovarian carcinoma, accounting for 10% of cases. Clear cell carcinoma typically presents with stage I or II disease, and in this setting prognostic markers could aid in management decisions, in particular the decision to treat with adjuvant chemotherapy. We tested whether expression of insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3, also known as IMP3) can serve as a new biomarker to predict outcome for patients with clear cell carcinoma and other subtypes of ovarian carcinoma. The expression of IGF2BP3 was evaluated by immunohistochemistry in 475 ovarian carcinomas of different subtypes and correlated with disease-specific survival. IGF2BP3 antibody specificity was validated by correlation of IGF2BP3 protein with mRNA expression level in a series of 35 ovarian carcinomas (r=0.849, P<0.0001). IGF2BP3 protein expression was an independent marker of reduced disease-specific survival (risk ratio 2.9, 95% confidence interval 1.4-5.8) in the clear cell subtype (N=128), but not in high-grade serous (N=198) or endometrioid (N=121) carcinomas. The prognostic significance of IGF2BP3 expression for reduced disease specific survival (risk ratio 2.6, 95% confidence interval 1.3-5.0) was confirmed in an independent series of cases (N=150) from three different centers in North America. We conclude that IGF2BP3 is the first biomarker of prognostic significance in ovarian clear cell carcinoma that has been validated in an independent case series. PMID- 19136933 TI - Increased TRPC3 expression in vascular endothelium of patients with malignant hypertension. AB - An increased expression of transient receptor potential canonical type 3 (TRPC3) cation channels has been proposed as one of the factors contributing to the pathogenesis of hypertension. To test that hypothesis we compared the expression of TRPC3 and TRPC6 as an endogenous control in human vascular endothelium of preglomerular arterioles in kidney biopsies from six patients with malignant hypertension and from four patients with diarrhea-associated hemolytic-uremic syndrome. Patients with malignant hypertension showed significantly higher systolic blood pressure and more prominent expression of TRPC3 in vascular endothelium of preglomerular arterioles compared to patients with hemolytic uremic syndrome. The expression of TRPC6 was not different between the two groups. The study supports the hypothesis that the increased expression of TRPC3 is associated with malignant hypertension in humans. PMID- 19136934 TI - REG4 is associated with carcinogenesis in the 'intestinal' pathway of intraductal papillary mucinous neoplasms. AB - Subclassification of intraductal papillary mucinous neoplasms of the pancreas (IPMNs), based on morphological features and immunohistochemical profiles, has been proposed. Intestinal-type IPMNs frequently show moderate to severe dysplasia. Regenerating islet-derived family, member 4 (REG4) is associated with the adenoma-carcinoma sequence in colon cancer and it is also associated with intestinal phenotype. Therefore, to identify REG4 expression in IPMNs may be helpful to detect high-grade IPMNs. We also investigated REG4 expression and CDX2 expression in IPMNs. To investigate the expressions of REG4 and CDX2 in IPMNs and in invasive ductal adenocarcinoma derived from IPMN, we used immunohistochemical staining and microdissection-based quantitative real-time reverse transcription polymerase chain reaction. Among 125 IPMNs, 43 (34%) were positive for REG4 and most of the intestinal-type IPMNs showed its expression (35/38). The positive ratio of REG4 expression in colloid carcinoma (5/7) was significantly higher than that in tubular carcinoma (1/17; P=0.003). Most of CDX2-positive cases (31/33) expressed REG4 protein, whereas only 12 of 92 CDX2-negative cases did (P<0.001). The levels of REG4 mRNA in intestinal-type IPMN were significantly higher compared to those in gastric-type IPMN or to normal pancreatic ductal epithelium (P=0.005, P=0.004, respectively). REG4 expression was observed more frequently in borderline lesions (14/28) and carcinoma (21/45) compared to adenoma (8/52). Using the Ki-67 labeling index, REG4 expression was significantly correlated with proliferative activity in borderline lesions. We conclude that REG4 is involved in the 'intestinal' pathway of carcinogenesis in IPMN. PMID- 19136935 TI - Endometrial changes from short-term therapy with CDB-4124, a selective progesterone receptor modulator. AB - Selective progesterone receptor modulators are a class of drugs with progesterone antagonist activity that may confer therapeutic benefit for reproductive disorders in premenopausal women. Endometrial structure, which is dynamically controlled by circulating sex hormones, is likely to be perturbed by progesterone receptor modulators through their progesterone antagonist properties. We examined endometrial histology in 58 premenopausal women treated with the progesterone receptor modulator CDB-4124 (also known as Proellex) for endometriosis or uterine leiomyomata in two clinical trials. Endometrial biopsies obtained after 3 or 6 months with doses of 12.5, 25, or 50 mg daily oral CDB-4124 were reviewed independently by three pathologists. Consensus diagnoses using the World Health Organization hyperplasia scoring system, comments on specific histologic features, and clinical annotation were collected and analyzed. The majority of the endometrial biopsies (103 of 174 biopsies) contained histologic changes that are not seen during normal menstrual cycles. The histology of CDB-4124-treated patients was generally inactive or atrophic, and less frequently, proliferative or secretory, superimposed upon which were novel changes including formation of cystically dilated glands, and secretory changes coexisting with mitoses and apoptotic bodies. With increasing treatment dose and duration, the cysts became predominant and their lining inactive or atrophic. Cystic glands in the CDB-4124 treated subjects correlated with increased endometrial thickness by ultrasound. None of the CDB-4124-treated patients developed endometrial carcinoma or hyperplasia while on therapy. CDB-4124 therapy for 3-6 months produces histologic changes that are sufficiently novel that they might easily be misinterpreted by pathologists, particularly as disordered proliferative or hyperplastic endometrium. Knowledge of the constellation of endometrial changes associated with this agent and other progesterone receptor modulators, including cystic architecture and mixed non-physiologic epithelial changes will prevent misdiagnosis. PMID- 19136936 TI - Chemokine receptors CXCR4 and CCR7 promote metastasis by preventing anoikis in cancer cells. AB - Chemokine receptors are essential mediators of the metastatic spread in various cancer types; however their precise function in the development of secondary tumors remains poorly understood. We report here a novel property of the chemokine receptors CXCR4 and CCR7 in inhibiting detachment-induced cell death- anoikis, which is believed to be one of the major blocks in the metastatic spread of various neoplasms. Activation of these chemokine receptors by their respective ligands, CXCL12 and CCL21 specifically reduced the sensitivity of metastatic breast cancer cells to anoikis by a distinct mechanism of selective regulation of pro-apoptotic Bmf and anti-apoptotic Bcl-xL proteins. Consequently, functional CXCR4 and CCR7 increased cell survival in the absence of correct ECM attachment both in vitro and in vivo. We also demonstrated that preventing chemokine-induced reduction in Bmf levels significantly attenuated breast cancer metastasis in an experimental mouse model. These results provide evidence for a previously unknown axis in malignant tumors, which connects chemokine receptors with deregulated apoptosis in the absence of the appropriate cell--ECM interaction and may offer novel targets for therapeutic intervention for the treatment of metastatic breast and potentially other tumors. PMID- 19136937 TI - Dynamics of outer mitochondrial membrane permeabilization during apoptosis. AB - Individual cells within a population undergo apoptosis at distinct, apparently random time points. By analyzing cellular mitotic history, we identified that sibling HeLa cell pairs, in contrast to random cell pairs, underwent apoptosis synchronously. This allowed us to use high-speed cellular imaging to investigate mitochondrial outer membrane permeabilization (MOMP), a highly coordinated, rapid process during apoptosis, at a temporal resolution approximately 100 times higher than possible previously. We obtained new functional and mechanistic insight into the process of MOMP: We were able to determine the kinetics of pore formation in the outer mitochondrial membrane from the initiation phase of cytochrome-c-GFP redistribution, and showed differential pore formation kinetics in response to intrinsic or extrinsic apoptotic stimuli (staurosporine, tumor necrosis factor related apoptosis-inducing ligand (TRAIL)). We also detected that the onset of mitochondrial permeabilization frequently proceeded as a wave through the cytosol, and that the frequency of wave occurrence in response to TRAIL was reduced by inhibition of protein kinase CK2. Computational analysis by a partial differential equation model suggested that the spread of permeabilization signals could sufficiently be explained by diffusion-adsorption velocities of locally generated permeabilization inducers. Taken together, our study yielded the first comprehensive analysis of clonal cell-to-cell variability in apoptosis execution and allowed to visualize and explain the dynamics of MOMP in cells undergoing apoptosis. PMID- 19136938 TI - C-terminal binding protein maintains mitochondrial activities. AB - Mitochondria are essential organelles that are responsible for cellular energy production and cell death in response to various stimuli. Although C-terminal binding protein (CtBP) functions as a metabolic sensor in transcriptional corepressor complex, it is unclear whether CtBP controls gene transcription in response to metabolic stress. In this study, we found that CtBP represses Bcl-2 associated X protein (Bax) transcription in glucose-rich media by binding to the E-box region of the Bax promoter. Glucose withdrawal leads to the dissociation of CtBP from the Bax promoter and significant changes of the histone codes in the Bax promoter. CtBP knockout increases Bax transcription, ablates mitochondrial morphology and reduces mitochondrial activities. Ectopic expression of CtBP or knockdown of Bax in ctbp-knockout cells recovers mitochondrial morphology and function, suggesting that CtBP functions as a metabolic sensor that maintains mitochondrial activities. Our findings provide insights into how the intracellular energy level is reflected into gene transcription involved in mitochondrial morphology and function. PMID- 19136939 TI - Visualizing CTL activity for different CD8+ effector T cells supports the idea that lower TCR/epitope avidity may be advantageous for target cell killing. AB - Time-lapse video microscopy allows analysis of the interaction between individual CTLs and adherent peptide-pulsed targets, from contact, to lymphocyte detachment, APC rounding, phosphatidylserine exposure and finally loss of plasma membrane integrity characteristic of end-stage apoptosis. Using in vitro-stimulated effectors specific for the ovalbumin K(b)OVA(257) (OT-I) and influenza A virus D(b)NP(366) and D(b)PA(224) epitopes, no significant correlation was found between the duration of CTL contact and the time to phosphatidylserine exposure or loss of membrane integrity. Furthermore, there were minimal indications that transgenic T cells specific for the K(b)OVA(257) epitope (TCR) diversity had any effect. However, when the analysis was repeated with D(b)NP(366) and D(b)PA(224) specific CTLs recovered directly from the lungs of mice with influenza pneumonia, the lower avidity D(b)NP(366)-specific set was found to elute much more quickly. Shorter contact time may allow individual CTLs to lyse more targets, suggesting that lower TCR/epitope avidity may be more beneficial than higher epitope avidity for cell-mediated immunity. PMID- 19136940 TI - GAP-43 is essential for the neurotrophic effects of BDNF and positive AMPA receptor modulator S18986. AB - Positive alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor modulators include benzamide compounds that allosterically modulate AMPA glutamate receptors. These small molecules that cross the blood-brain barrier have been shown to act as a neuroprotectant by increasing the levels of endogenous brain-derived neurotrophic factor (BDNF). Positive AMPA receptor modulators have also been shown to increase the levels of growth-associated protein-43 (GAP-43). GAP-43 plays a major role in many aspects of neuronal function in vertebrates. The goal of this study was to determine whether GAP-43 was important in mediating the actions of positive AMPA receptor modulator (S18986) and BDNF. Using cortical cultures from GAP-43 knockout and control mice, we show that (1) GAP-43 is upregulated in response to S18986 and BDNF in control cultures; (2) this upregulation of GAP-43 is essential for mediating the neuroprotective effects of S18986 and BDNF; (3) administration of S18986 and BDNF leads to an increase in the expression of the glutamate transporters GLT-1 and GLAST that are key to limiting excitotoxic cell death and this increase in GLT-1 and GLAST expression is completely blocked in the absence of GAP-43. Taken together this study concludes that GAP-43 is an important mediator of the neurotrophic effects of S18986 and BDNF on neuronal survival and plasticity, and is essential for the success of positive AMPA receptor modulator-BDNF-based neurotrophin therapy. PMID- 19136941 TI - The role of Bcl-2 family member BNIP3 in cell death and disease: NIPping at the heels of cell death. AB - Bcl-2 nineteen-kilodalton interacting protein (BNIP3) is a BH-3-only Bcl-2 family member whose expression levels increase during stress such as hypoxia through hypoxia-inducing factor-1-dependent or -independent mechanisms. When BNIP3 expression is induced, it localizes to the mitochondria and triggers a loss of membrane potential, and an increase in the reactive oxygen species production, which often leads to cell death. Cells under normal growth conditions suppress BNIP3 expression through transcriptional repression. There is considerable debate in the literature regarding what type of cell death is induced by BNIP3. It has been observed that BNIP3 could induce necrosis, autophagy and/or apoptosis. In contrast, other studies indicate that BNIP3 could promote cell survival. Besides its cell death regulation, BNIP3 plays a key role in the pathogenicity of many diseases. In cardiac infarction, loss of BNIP3 expression has been shown to reduce the number of damaged cardiomyocytes after ischemia and reperfusion. BNIP3 expression also plays an important role in the deregulation of cell death in many cancers. In this review, we will discuss the different and often contradictory mechanisms of BNIP3 regulation of cell death and the role that BNIP3 may play in diseases. PMID- 19136942 TI - TGF-beta induces apoptosis in human B cells by transcriptional regulation of BIK and BCL-XL. AB - Transforming growth factor-beta (TGF-beta) potently induces apoptosis in Burkitt's lymphoma (BL) cell lines and in explanted primary human B lymphocytes. The physiological relevance and mechanism of TGF-beta-mediated apoptosis induction in these cells remains to be determined. Here we demonstrate the requirement for TGF-beta-mediated regulation of BIK and BCL-X(L) to activate an intrinsic apoptotic pathway in centroblastic BL cells. TGF-beta directly induced transcription of BIK and a consensus Smad-binding element identified in the BIK promoter recruits TGF-beta-activated Smad transcription factor complexes in vivo. TGF-beta also transcriptionally repressed expression of the apoptosis inhibitor BCL-X(L). Inhibition of BCL-X(L) sensitised BL cells to TGF-beta-induced apoptosis whereas overexpression of BCL-X(L) or suppression of BIK by shRNA, diminished TGF-beta-induced apoptosis. BIK and BCL-X(L) were also identified as TGF-beta target genes in purified normal human centroblast B cells and immunohistochemical analyses of tonsil tissue revealed widespread TGF-beta receptor-regulated Smad activation and a focal pattern of BIK expression. Furthermore, using a selective inhibitor of the TGF-beta receptor we provide evidence that autocrine TGF-beta signalling through ALK5 contributes to the default apoptotic programme in normal human centroblasts undergoing spontaneous apoptosis. Our data suggests that TGF-beta may act as a physiological mediator of human germinal centre homoeostasis by regulation of BIK and BCL-X(L). PMID- 19136943 TI - Transcriptome sequencing to detect gene fusions in cancer. AB - Recurrent gene fusions, typically associated with haematological malignancies and rare bone and soft-tissue tumours, have recently been described in common solid tumours. Here we use an integrative analysis of high-throughput long- and short read transcriptome sequencing of cancer cells to discover novel gene fusions. As a proof of concept, we successfully used integrative transcriptome sequencing to 're-discover' the BCR-ABL1 (ref. 10) gene fusion in a chronic myelogenous leukaemia cell line and the TMPRSS2-ERG gene fusion in a prostate cancer cell line and tissues. Additionally, we nominated, and experimentally validated, novel gene fusions resulting in chimaeric transcripts in cancer cell lines and tumours. Taken together, this study establishes a robust pipeline for the discovery of novel gene chimaeras using high-throughput sequencing, opening up an important class of cancer-related mutations for comprehensive characterization. PMID- 19136944 TI - Genetic architecture of mouse skin inflammation and tumour susceptibility. AB - Germline polymorphisms in model organisms and humans influence susceptibility to complex trait diseases such as inflammation and cancer. Mice of the Mus spretus species are resistant to tumour development, and crosses between M. spretus and susceptible Mus musculus strains have been used to map locations of genetic variants that contribute to skin cancer susceptibility. We have integrated germline polymorphisms with gene expression in normal skin from a M. musculus x M. spretus backcross to generate a network view of the gene expression architecture of mouse skin. Here we demonstrate how this approach identifies expression motifs that contribute to tissue organization and biological functions related to inflammation, haematopoiesis, cell cycle control and tumour susceptibility. Motifs associated with inflammation, epidermal barrier function and proliferation are differentially regulated in backcross mice susceptible or resistant to tumour development. The intestinal stem cell marker Lgr5 is identified as a candidate master regulator of the hair follicle, and the vitamin D receptor (Vdr) is linked to coordinated control of epidermal barrier function, inflammation and tumour susceptibility. PMID- 19136946 TI - Polymer chain dynamics and glass transition in athermal polymer/nanoparticle mixtures. AB - Polymer nanocomposites (PNCs), prepared by incorporating nanoparticles within a polymer host, generally exhibit properties that differ significantly from those of the host, even with small amounts of nanoparticles. A significant challenge is how to tailor the properties of these materials for applications (structural and biomedical to optoelectronic), because PNCs derive their properties from a collective and complex range of entropic and enthalpic interactions. Here, we show that PNCs, prepared from athermal mixtures of polymer-chain-grafted gold nanoparticles and unentangled polymer chains, may exhibit increases or decreases in their relaxation dynamics, and viscosity, by over an order of magnitude through control of nanoparticle concentration, nanoparticle size, grafting density and grafting chain degree of polymerization. In addition, we show how the glass transition may also be tailored by up to 10 degrees with the addition of less than 1.0 wt% nanoparticles to the polymer host. PMID- 19136945 TI - Adaptive immune features of natural killer cells. AB - In an adaptive immune response, naive T cells proliferate during infection and generate long-lived memory cells that undergo secondary expansion after a repeat encounter with the same pathogen. Although natural killer (NK) cells have traditionally been classified as cells of the innate immune system, they share many similarities with cytotoxic T lymphocytes. We use a mouse model of cytomegalovirus infection to show that, like T cells, NK cells bearing the virus specific Ly49H receptor proliferate 100-fold in the spleen and 1,000-fold in the liver after infection. After a contraction phase, Ly49H-positive NK cells reside in lymphoid and non-lymphoid organs for several months. These self-renewing 'memory' NK cells rapidly degranulate and produce cytokines on reactivation. Adoptive transfer of these NK cells into naive animals followed by viral challenge results in a robust secondary expansion and protective immunity. These findings reveal properties of NK cells that were previously attributed only to cells of the adaptive immune system. PMID- 19136947 TI - Infection-mimicking materials to program dendritic cells in situ. AB - Cancer vaccines typically depend on cumbersome and expensive manipulation of cells in the laboratory, and subsequent cell transplantation leads to poor lymph node homing and limited efficacy. We propose that materials mimicking key aspects of bacterial infection may instead be used to directly control immune-cell trafficking and activation in the body. It is demonstrated that polymers can be designed to first release a cytokine to recruit and house host dendritic cells, and subsequently present cancer antigens and danger signals to activate the resident dendritic cells and markedly enhance their homing to lymph nodes. Specific and protective anti-tumour immunity was generated with these materials, as 90% survival was achieved in animals that otherwise die from cancer within 25 days. These materials show promise as cancer vaccines, and more broadly suggest that polymers may be designed to program and control the trafficking of a variety of cell types in the body. PMID- 19136948 TI - Complex and hierarchical micelle architectures from diblock copolymers using living, crystallization-driven polymerizations. AB - Block copolymers consist of two or more chemically distinct polymer segments, or blocks, connected by a covalent link. In a selective solvent for one of the blocks, core-corona micelle structures are formed. We demonstrate that living polymerizations driven by the epitaxial crystallization of a core-forming metalloblock represent a synthetic tool that can be used to generate complex and hierarchical micelle architectures from diblock copolymers. The use of platelet micelles as initiators enables the formation of scarf-like architectures in which cylindrical micelle tassels of controlled length are grown from specific crystal faces. A similar process enables the fabrication of brushes of cylindrical micelles on a crystalline homopolymer substrate. Living polymerizations driven by heteroepitaxial growth can also be accomplished and are illustrated by the formation of tri- and pentablock and scarf architectures with cylinder-cylinder and platelet-cylinder connections, respectively, that involve different core forming metalloblocks. PMID- 19136949 TI - Genetic variation in PCDH11X is associated with susceptibility to late-onset Alzheimer's disease. AB - By analyzing late-onset Alzheimer's disease (LOAD) in a genome-wide association study (313,504 SNPs, three series, 844 cases and 1,255 controls) and evaluating the 25 SNPs with the most significant allelic association in four additional series (1,547 cases and 1,209 controls), we identified a SNP (rs5984894) on Xq21.3 in PCDH11X that is strongly associated with LOAD in individuals of European descent from the United States. Analysis of rs5984894 by multivariable logistic regression adjusted for sex gave global P values of 5.7 x 10(-5) in stage 1, 4.8 x 10(-6) in stage 2 and 3.9 x 10(-12) in the combined data. Odds ratios were 1.75 (95% CI = 1.42-2.16) for female homozygotes (P = 2.0 x 10(-7)) and 1.26 (95% CI = 1.05-1.51) for female heterozygotes (P = 0.01) compared to female noncarriers. For male hemizygotes (P = 0.07) compared to male noncarriers, the odds ratio was 1.18 (95% CI = 0.99-1.41). PMID- 19136950 TI - Increased LIS1 expression affects human and mouse brain development. AB - Deletions of the PAFAH1B1 gene (encoding LIS1) in 17p13.3 result in isolated lissencephaly sequence, and extended deletions including the YWHAE gene (encoding 14-3-3epsilon) cause Miller-Dieker syndrome. We identified seven unrelated individuals with submicroscopic duplication in 17p13.3 involving the PAFAH1B1 and/or YWHAE genes, and using a 'reverse genomics' approach, characterized the clinical consequences of these duplications. Increased PAFAH1B1 dosage causes mild brain structural abnormalities, moderate to severe developmental delay and failure to thrive. Duplication of YWHAE and surrounding genes increases the risk for macrosomia, mild developmental delay and pervasive developmental disorder, and results in shared facial dysmorphologies. Transgenic mice conditionally overexpressing LIS1 in the developing brain showed a decrease in brain size, an increase in apoptotic cells and a distorted cellular organization in the ventricular zone, including reduced cellular polarity but preserved cortical cell layer identity. Collectively, our results show that an increase in LIS1 expression in the developing brain results in brain abnormalities in mice and humans. PMID- 19136951 TI - Identification of a putative lysosomal cobalamin exporter altered in the cblF defect of vitamin B12 metabolism. AB - Vitamin B(12) (cobalamin) is essential in animals for metabolism of branched chain amino acids and odd chain fatty acids, and for remethylation of homocysteine to methionine. In the cblF inborn error of vitamin B(12) metabolism, free vitamin accumulates in lysosomes, thus hindering its conversion to cofactors. Using homozygosity mapping in 12 unrelated cblF individuals and microcell-mediated chromosome transfer, we identified a candidate gene on chromosome 6q13, LMBRD1, encoding LMBD1, a lysosomal membrane protein with homology to lipocalin membrane receptor LIMR. We identified five different frameshift mutations in LMBRD1 resulting in loss of LMBD1 function, with 18 of the 24 disease chromosomes carrying the same mutation embedded in a common 1.34 Mb haplotype. Transfection of fibroblasts of individuals with cblF with wild-type LMBD1 rescued cobalamin coenzyme synthesis and function. This work identifies LMBRD1 as the gene underlying the cblF defect of cobalamin metabolism and suggests that LMBD1 is a lysosomal membrane exporter for cobalamin. PMID- 19136952 TI - DCTN1 mutations in Perry syndrome. AB - Perry syndrome consists of early-onset parkinsonism, depression, severe weight loss and hypoventilation, with brain pathology characterized by TDP-43 immunostaining. We carried out genome-wide linkage analysis and identified five disease-segregating mutations affecting the CAP-Gly domain of dynactin (encoded by DCTN1) in eight families with Perry syndrome; these mutations diminish microtubule binding and lead to intracytoplasmic inclusions. Our findings show that DCTN1 mutations, previously associated with motor neuron disease, can underlie the selective vulnerability of other neuronal populations in distinct neurodegenerative disorders. PMID- 19136953 TI - 15q13.3 microdeletions increase risk of idiopathic generalized epilepsy. AB - We identified 15q13.3 microdeletions encompassing the CHRNA7 gene in 12 of 1,223 individuals with idiopathic generalized epilepsy (IGE), which were not detected in 3,699 controls (joint P = 5.32 x 10(-8)). Most deletion carriers showed common IGE syndromes without other features previously associated with 15q13.3 microdeletions, such as intellectual disability, autism or schizophrenia. Our results indicate that 15q13.3 microdeletions constitute the most prevalent risk factor for common epilepsies identified to date. PMID- 19136954 TI - Experimental evolution reveals natural selection on standing genetic variation. AB - Evolution depends on genetic variation generated by mutation or recombination from standing genetic variation. In sexual organisms, little is known about the molecular population genetics of adaptation and reverse evolution. We carry out 50 generations of experimental reverse evolution in populations of Drosophila melanogaster, previously differentiated by forward evolution, and follow changes in the frequency of SNPs in both arms of the third chromosome. We characterize the effects of sampling finite population sizes and natural selection at the genotype level. We demonstrate that selection has occurred at several loci and further that there is no general loss or gain of allele diversity. We also observe that despite the complete convergence to ancestral levels of adaptation, allele frequencies only show partial return. PMID- 19136955 TI - An RNA code for the FOX2 splicing regulator revealed by mapping RNA-protein interactions in stem cells. AB - The elucidation of a code for regulated splicing has been a long-standing goal in understanding the control of post-transcriptional gene expression events that are crucial for cell survival, differentiation and development. We decoded functional RNA elements in vivo by constructing an RNA map for the cell type-specific splicing regulator FOX2 (also known as RBM9) via cross-linking immunoprecipitation coupled with high-throughput sequencing (CLIP-seq) in human embryonic stem cells. The map identified a large cohort of specific FOX2 targets, many of which are themselves splicing regulators, and comparison between the FOX2 binding profile and validated splicing events revealed a general rule for FOX2 regulated exon inclusion or skipping in a position-dependent manner. These findings suggest that FOX2 functions as a critical regulator of a splicing network, and we further show that FOX2 is important for the survival of human embryonic stem cells. PMID- 19136956 TI - SRS2 and SGS1 prevent chromosomal breaks and stabilize triplet repeats by restraining recombination. AB - Several molecular mechanisms have been proposed to explain trinucleotide repeat expansions. Here we show that in yeast srs2Delta cells, CTG repeats undergo both expansions and contractions, and they show increased chromosomal fragility. Deletion of RAD52 or RAD51 suppresses these phenotypes, suggesting that recombination triggers trinucleotide repeat instability in srs2Delta cells. In sgs1Delta cells, CTG repeats undergo contractions and increased fragility by a mechanism partially dependent on RAD52 and RAD51. Analysis of replication intermediates revealed abundant joint molecules at the CTG repeats during S phase. These molecules migrate similarly to reversed replication forks, and their presence is dependent on SRS2 and SGS1 but not RAD51. Our results suggest that Srs2 promotes fork reversal in repetitive sequences, preventing repeat instability and fragility. In the absence of Srs2 or Sgs1, DNA damage accumulates and is processed by homologous recombination, triggering repeat rearrangements. PMID- 19136957 TI - Replisome stalling and stabilization at CGG repeats, which are responsible for chromosomal fragility. AB - Expanded CGG repeats cause chromosomal fragility and hereditary neurological disorders in humans. Replication forks stall at CGG repeats in a length-dependent manner in primate cells and in yeast. Saccharomyces cerevisiae proteins Tof1 and Mrc1 facilitate replication fork progression through CGG repeats. Remarkably, the fork-stabilizing role of Mrc1 does not involve its checkpoint function. Thus, chromosomal fragility might occur when forks stalled at expanded CGG repeats escape the S-phase checkpoint. PMID- 19136958 TI - Structures of endonuclease V with DNA reveal initiation of deaminated adenine repair. AB - Endonuclease V (EndoV) initiates a major base-repair pathway for nitrosative deamination resulting from endogenous processes and increased by oxidative stress from mitochondrial dysfunction or inflammatory responses. We solved the crystal structures of Thermotoga maritima EndoV in complex with a hypoxanthine lesion substrate and with product DNA. The PYIP wedge motif acts as a minor groove damage sensor for helical distortions and base mismatches and separates DNA strands at the lesion. EndoV incises DNA with an unusual offset nick 1 nucleotide 3' of the lesion, as the deaminated adenine is rotated approximately 90 degrees into a recognition pocket approximately 8 A from the catalytic site. Tight binding by the lesion-recognition pocket in addition to Mg(2+) and hydrogen bonding interactions to the DNA ends stabilize the product complex, suggesting an orderly recruitment of downstream proteins in this base-repair pathway. PMID- 19136959 TI - High-resolution dynamic mapping of histone-DNA interactions in a nucleosome. AB - The nature of the nucleosomal barrier that regulates access to the underlying DNA during many cellular processes is not fully understood. Here we present a detailed map of histone-DNA interactions along the DNA sequence to near base pair accuracy by mechanically unzipping single molecules of DNA, each containing a single nucleosome. This interaction map revealed a distinct approximately 5-bp periodicity that was enveloped by three broad regions of strong interactions, with the strongest occurring at the dyad and the other two about +/-40-bp from the dyad. Unzipping up to the dyad allowed recovery of a canonical nucleosome upon relaxation of the DNA, but unzipping beyond the dyad resulted in removal of the histone octamer from its initial DNA sequence. These findings have important implications for how RNA polymerase and other DNA-based enzymes may gain access to DNA associated with a nucleosome. PMID- 19136960 TI - Interleukin 7 signaling in dendritic cells regulates the homeostatic proliferation and niche size of CD4+ T cells. AB - Interleukin 7 (IL-7) and T cell antigen receptor signals have been proposed to be the main drivers of homeostatic T cell proliferation. However, it is not known why CD4(+) T cells undergo less-efficient homeostatic proliferation than CD8(+) T cells do. Here we show that systemic IL-7 concentrations increased during lymphopenia because of diminished use of IL-7 but that IL-7 signaling on IL-7 receptor-alpha-positive (IL-7Ralpha(+)) dendritic cells (DCs) in lymphopenic settings paradoxically diminished the homeostatic proliferation of CD4(+) T cells. This effect was mediated at least in part by IL-7-mediated downregulation of the expression of major histocompatibility complex class II on IL-7Ralpha(+) DCs. Our results indicate that IL-7Ralpha(+) DCs are regulators of the peripheral CD4(+) T cell niche and that IL-7 signals in DCs prevent uncontrolled CD4(+) T cell population expansion in vivo. PMID- 19136961 TI - Regulation of conformer-specific activation of the integrin LFA-1 by a chemokine triggered Rho signaling module. AB - Regulation of the affinity of the beta(2) integrin LFA-1 by chemokines is critical to lymphocyte trafficking, but the signaling mechanisms that control this process are not well understood. Here we investigated the signaling events controlling LFA-1 affinity triggering by chemokines in human primary T lymphocytes. We found that the small GTPase Rac1 mediated chemokine-induced LFA-1 affinity triggering and lymphocyte arrest in high endothelial venules. Unexpectedly, another Rho family member, Cdc42, negatively regulated LFA-1 activation. The Rho effectors PLD1 and PIP5KC were also critical to LFA-1 affinity modulation. Notably, PIP5KC was found to specifically control the transition of LFA-1 from an extended low-intermediate state to a high-affinity state, which correlated with lymphocyte arrest. Thus, chemokines control lymphocyte trafficking by triggering a Rho-dependent signaling cascade leading to conformer-specific modulation of LFA-1 affinity. PMID- 19136962 TI - Foxo1 links homing and survival of naive T cells by regulating L-selectin, CCR7 and interleukin 7 receptor. AB - Foxo transcription factors have a conserved role in the adaptation of cells and organisms to nutrient and growth factor availability. Here we show that Foxo1 has a crucial, nonredundant role in T cells. In naive T cells, Foxo1 controlled the expression of the adhesion molecule L-selectin, the chemokine receptor CCR7 and the transcription factor Klf2, and its deletion was sufficient to alter lymphocyte trafficking. Furthermore, Foxo1 deficiency resulted in a severe defect in interleukin 7 receptor alpha-chain (IL-7Ralpha) expression associated with its ability to bind an Il7r enhancer. Finally, growth factor withdrawal induced a Foxo1-dependent increase in Sell, Klf2 and Il7r expression. These data suggest that Foxo1 regulates the homeostasis and life span of naive T cells by sensing growth factor availability and regulating homing and survival signals. PMID- 19136963 TI - Loss of ETHE1, a mitochondrial dioxygenase, causes fatal sulfide toxicity in ethylmalonic encephalopathy. AB - Ethylmalonic encephalopathy is an autosomal recessive, invariably fatal disorder characterized by early-onset encephalopathy, microangiopathy, chronic diarrhea, defective cytochrome c oxidase (COX) in muscle and brain, high concentrations of C4 and C5 acylcarnitines in blood and high excretion of ethylmalonic acid in urine. ETHE1, a gene encoding a beta-lactamase-like, iron-coordinating metalloprotein, is mutated in ethylmalonic encephalopathy. In bacteria, ETHE1 like sequences are in the same operon of, or fused with, orthologs of TST, the gene encoding rhodanese, a sulfurtransferase. In eukaryotes, both ETHE1 and rhodanese are located within the mitochondrial matrix. We created a Ethe1(-/-) mouse that showed the cardinal features of ethylmalonic encephalopathy. We found that thiosulfate was excreted in massive amounts in urine of both Ethe1(-/-) mice and humans with ethylmalonic encephalopathy. High thiosulfate and sulfide concentrations were present in Ethe1(-/-) mouse tissues. Sulfide is a powerful inhibitor of COX and short-chain fatty acid oxidation, with vasoactive and vasotoxic effects that explain the microangiopathy in ethylmalonic encephalopathy patients. Sulfide is detoxified by a mitochondrial pathway that includes a sulfur dioxygenase. Sulfur dioxygenase activity was absent in Ethe1(-/-) mice, whereas it was markedly increased by ETHE1 overexpression in HeLa cells and Escherichia coli. Therefore, ETHE1 is a mitochondrial sulfur dioxygenase involved in catabolism of sulfide that accumulates to toxic levels in ethylmalonic encephalopathy. PMID- 19136964 TI - AdPLA ablation increases lipolysis and prevents obesity induced by high-fat feeding or leptin deficiency. AB - A main function of white adipose tissue is to release fatty acids from stored triacylglycerol for other tissues to use as an energy source. Whereas endocrine regulation of lipolysis has been extensively studied, autocrine and paracrine regulation is not well understood. Here we describe the role of the newly identified major adipocyte phospholipase A(2), AdPLA (encoded by Pla2g16, also called HREV107), in the regulation of lipolysis and adiposity. AdPLA-null mice have a markedly higher rate of lipolysis owing to increased cyclic AMP levels arising from the marked reduction in the amount of adipose prostaglandin E(2) that binds the Galpha(i)-coupled receptor, EP3. AdPLA-null mice have markedly reduced adipose tissue mass and triglyceride content but normal adipogenesis. They also have higher energy expenditure with increased fatty acid oxidation within adipocytes. AdPLA-deficient ob/ob mice remain hyperphagic but lean, with increased energy expenditure, yet have ectopic triglyceride storage and insulin resistance. AdPLA is a major regulator of adipocyte lipolysis and is crucial for the development of obesity. PMID- 19136965 TI - Reprogramming of fibroblasts into induced pluripotent stem cells with orphan nuclear receptor Esrrb. AB - The dominant effect of transcription factors in imparting expanded potency is best exemplified by the reprogramming of fibroblasts to pluripotent cells using retrovirus-mediated transduction of defined transcription factors. In the murine system, Oct4, Sox2, c-Myc and Klf4 are sufficient to convert fibroblasts to induced pluripotent stem (iPS) cells that have many characteristics of embryonic stem (ES) cells. Here we show that the orphan nuclear receptor Esrrb functions in conjunction with Oct4 and Sox2 to mediate reprogramming of mouse embryonic fibroblasts (MEFs) to iPS cells. Esrrb-reprogrammed cells share similar expression and epigenetic signatures as ES cells. These cells are also pluripotent and can differentiate in vitro and in vivo into the three major embryonic cell lineages. Furthermore, these cells contribute to mouse chimaeras and are germline transmissible. In ES cells, Esrrb targets many genes involved in self-renewal and pluripotency. This suggests that Esrrb may mediate reprogramming through the upregulation of ES-cell-specific genes. Our findings also indicate that it is possible to reprogram MEFs without exogenous Klf transcription factors and link a nuclear receptor to somatic cell reprogramming. PMID- 19136966 TI - Nuclear signalling by tumour-associated antigen EpCAM. AB - EpCAM was found to be overexpressed on epithelial progenitors, carcinomas and cancer-initiating cells. The role of EpCAM in proliferation, and its association with cancer is poorly explained by proposed cell adhesion functions. Here we show that regulated intramembrane proteolysis activates EpCAM as a mitogenic signal transducer in vitro and in vivo. This involves shedding of its ectodomain EpEX and nuclear translocation of its intracellular domain EpICD. Cleavage of EpCAM is sequentially catalysed by TACE and presenilin-2. Pharmacological inhibition or genetic silencing of either protease impairs growth-promoting signalling by EpCAM, which is compensated for by EpICD. Released EpICD associates with FHL2, beta-catenin and Lef-1 to form a nuclear complex that contacts DNA at Lef-1 consensus sites, induces gene transcription and is oncogenic in immunodeficient mice. In patients, EpICD was found in nuclei of colon carcinoma but not of normal tissue. Nuclear signalling of EpCAM explains how EpCAM functions in cell proliferation. PMID- 19136968 TI - Involvement of linear polyubiquitylation of NEMO in NF-kappaB activation. AB - Nuclear factor-kappaB (NF-kappaB) is a key transcription factor in inflammatory, anti-apoptotic and immune processes. The ubiquitin pathway is crucial in regulating the NF-kappaB pathway. We have found that the LUBAC ligase complex, composed of the two RING finger proteins HOIL-1L and HOIP, conjugates a head-to tail-linked linear polyubiquitin chain to substrates. Here, we demonstrate that LUBAC activates the canonical NF-kappaB pathway by binding to NEMO (NF-kappaB essential modulator, also called IKKgamma) and conjugates linear polyubiquitin chains onto specific Lys residues in the CC2-LZ domain of NEMO in a Ubc13 independent manner. Moreover, in HOIL-1 knockout mice and cells derived from these mice, NF-kappaB signalling induced by pro-inflammatory cytokines such as TNF-alpha and IL-1beta was suppressed, resulting in enhanced TNF-alpha-induced apoptosis in hepatocytes of HOIL-1 knockout mice. These results indicate that LUBAC is involved in the physiological regulation of the canonical NF-kappaB activation pathway through linear polyubiquitylation of NEMO. PMID- 19136967 TI - Interaction between TAK1-TAB1-TAB2 and RCAN1-calcineurin defines a signalling nodal control point. AB - The calcium-activated protein phosphatase calcineurin is controlled by regulator of calcineurin (RCAN) in organisms ranging from yeast to mammals. Here we performed a yeast two-hybrid screen with RCAN1 as bait, identifying TAK1 binding protein 2 (TAB2) as an interacting partner. TAB2 interacted directly with RCAN1 in vitro and in vivo, recruiting TAK1, TAB1 and calcineurin, forming a macromolecular signalling complex. Overexpression of TAK1 and TAB1, or active TAK1(DeltaN), promoted direct phosphorylation of RCAN1 in vitro and in vivo. TAK1 phosphorylated RCAN1 at Ser 94 and Ser 136, converting RCAN1 from an inhibitor to a facilitator of calcineurin-NFAT signalling, and enhancing NFATc1 nuclear translocation, NFAT transcriptional activation and the hypertrophic growth of cultured cardiomyocytes. The TAK1-TAB1-TAB2 and the calcineurin-NFAT signalling modules did not interact in Rcan1/2- or Tab2-deficient mouse embryonic fibroblast (MEF) cultures. Calcineurin activation also dephosphorylated and inhibited TAK1 and TAB1, an effect that was absent in Rcan1/2 deficient MEFs. Functionally, TAK1 was indispensable for the cardiomyocyte growth response induced by pro hypertrophic stimuli through calcineurin. These results describe a signalling relationship between two central regulatory pathways in which TAK1-TAB1-TAB2 selectively induces calcineurin-NFAT signalling through direct phosphorylation of RCAN1, while calcineurin activation diminishes TAK1 signalling by dephosphorylation of TAK1 and TAB1. PMID- 19136969 TI - Synaptotagmin IV: a multifunctional regulator of peptidergic nerve terminals. AB - Many members of the synaptotagmin (Syt) protein family bind Ca(2+) and trigger exocytosis, but some Syt proteins appear to have no Ca(2+)-dependent actions and their biological functions remain obscure. Syt IV is an activity-induced brain protein with no known Ca(2+)-dependent interactions and its subcellular localization and biological functions have sparked considerable controversy. We found Syt IV on both micro- and dense-core vesicles in posterior pituitary nerve terminals in mice. In terminals from Syt IV knockout mice compared with those from wild types, low Ca(2+) entry triggered more exocytosis, high Ca(2+) entry triggered less exocytosis and endocytosis was accelerated. In Syt IV knockouts, dense-core and microvesicle fusion was enhanced in cell-attached patches and dense-core vesicle fusion pores had conductances that were half as large as those in wild types. Given the neuroendocrine functions of the posterior pituitary, changes in Syt IV levels could be involved in endocrine transitions involving alterations in the release of the neuropeptides oxytocin and vasopressin. PMID- 19136970 TI - The tumor suppressor Pml regulates cell fate in the developing neocortex. AB - The control of cell fate in neural progenitor cells is critical for nervous system development. Nevertheless, the processes involved are only partially known. We found that the expression of the tumor suppressor Pml was restricted to neural progenitor cells (NPCs) in the developing neocortex of the mouse. Notably, in Pml(-/-) cortices, the overall number of proliferating NPCs was increased and transition between the two major progenitor types, radial glial cells and basal progenitors, was impaired. This in turn resulted in reduced differentiation and an overall decrease in the thickness of the cortex wall. In NPCs, Pml regulated the subcellular distribution of the retinoblastoma protein (pRb) and the protein phosphatase 1alpha, triggering pRb dephosphorylation. Together, these findings reveal an unexpected role of Pml in controlling the function of NPCs in the CNS. PMID- 19136971 TI - N-Acetylcysteine reverses cocaine-induced metaplasticity. AB - Cocaine addiction is characterized by an impaired ability to develop adaptive behaviors that can compete with cocaine seeking, implying a deficit in the ability to induce plasticity in cortico-accumbens circuitry crucial for regulating motivated behavior. We found that rats withdrawn from cocaine self administration had a marked in vivo deficit in the ability to develop long-term potentiation (LTP) and long-term depression (LTD) in the nucleus accumbens core subregion after stimulation of the prefrontal cortex. N-acetylcysteine (NAC) treatment prevents relapse in animal models and craving in humans by activating cystine-glutamate exchange and thereby stimulating extrasynaptic metabotropic glutamate receptors (mGluR). NAC treatment of rats restored the ability to induce LTP and LTD by indirectly stimulating mGluR2/3 and mGluR5, respectively. Our findings show that cocaine self-administration induces metaplasticity that inhibits further induction of synaptic plasticity, and this impairment can be reversed by NAC, a drug that also prevents relapse. PMID- 19136972 TI - Neural correlates of categorical perception in learned vocal communication. AB - The division of continuously variable acoustic signals into discrete perceptual categories is a fundamental feature of vocal communication, including human speech. Despite the importance of categorical perception to learned vocal communication, the neural correlates underlying this phenomenon await identification. We found that individual sensorimotor neurons in freely behaving swamp sparrows expressed categorical auditory responses to changes in note duration, a learned feature of their songs, and that the neural response boundary accurately predicted the categorical perceptual boundary measured in field studies of the same sparrow population. Furthermore, swamp sparrow populations that learned different song dialects showed different categorical perceptual boundaries that were consistent with the boundary being learned. Our results extend the analysis of the neural basis of perceptual categorization into the realm of vocal communication and advance the learned vocalizations of songbirds as a model for investigating how experience shapes categorical perception and the activity of categorically responsive neurons. PMID- 19136973 TI - Neuronal release of proBDNF. AB - Pro-brain-derived neurotrophic factor (proBDNF) and mature BDNF utilize distinct receptors to mediate divergent neuronal actions. Using new tools to quantitate endogenous BDNF isoforms, we found that mouse neurons secrete both proBDNF and mature BDNF. The highest levels of proBDNF and p75 were observed perinatally and declined, but were still detectable, in adulthood. Thus, BDNF actions are developmentally regulated by secretion of proBDNF or mature BDNF and by local expression of p75 and TrkB. PMID- 19136974 TI - Identification of distinct telencephalic progenitor pools for neuronal diversity in the amygdala. AB - The development of the amygdala, a central structure of the limbic system, remains poorly understood. We found that two spatially distinct and early specified telencephalic progenitor pools marked by the homeodomain transcription factor Dbx1 are major sources of neuronal cell diversity in the mature mouse amygdala. We found that Dbx1-positive cells of the ventral pallium generate the excitatory neurons of the basolateral complex and cortical amygdala nuclei. Moreover, Dbx1-derived cells comprise a previously unknown migratory stream that emanates from the preoptic area (POA), a ventral telencephalic domain adjacent to the diencephalic border. The Dbx1-positive, POA-derived population migrated specifically to the amygdala and, as defined by both immunochemical and electrophysiological criteria, generated a unique subclass of inhibitory neurons in the medial amygdala nucleus. Thus, this POA-derived population represents a previously unknown progenitor pool dedicated to the limbic system. PMID- 19136975 TI - Design of isoform-selective phospholipase D inhibitors that modulate cancer cell invasiveness. AB - Phospholipase D (PLD) is an essential enzyme responsible for the production of the lipid second messenger phosphatidic acid. Phosphatidic acid participates in both G protein-coupled receptor and receptor tyrosine kinase signal transduction networks. The lack of potent and isoform-selective inhibitors has limited progress in defining the cellular roles of PLD. We used a diversity-oriented synthetic approach and developed a library of PLD inhibitors with considerable pharmacological characterization. Here we report the rigorous evaluation of that library, which contains highly potent inhibitors, including the first isoform selective PLD inhibitors. Specific members of this series inhibit isoforms with >100-fold selectivity both in vitro and in cells. A subset of inhibitors was shown to block invasiveness in metastatic breast cancer models. These findings demonstrate the power of diversity-oriented synthesis combined with biochemical assays and mass spectrometric lipid profiling of cellular responses to develop the first isoform-selective PLD inhibitors--a new class of antimetastatic agents. PMID- 19136976 TI - Monomeric fluorescent timers that change color from blue to red report on cellular trafficking. AB - Based on the mechanism for chromophore formation in red fluorescent proteins, we developed three mCherry-derived monomeric variants, called fluorescent timers (FTs), that change their fluorescence from the blue to red over time. These variants exhibit distinctive fast, medium and slow blue-to-red chromophore maturation rates that depend on the temperature. At 37 degrees C, the maxima of the blue fluorescence are observed at 0.25, 1.2 and 9.8 h for the purified fast FT, medium-FT and slow-FT, respectively. The half-maxima of the red fluorescence are reached at 7.1, 3.9 and 28 h, respectively. The FTs show similar timing behavior in bacteria, insect and mammalian cells. Medium-FT allowed for tracking of the intracellular dynamics of the lysosome-associated membrane protein type 2A (LAMP-2A) and determination of its age in the targeted compartments. The results indicate that LAMP-2A transport through the plasma membrane and early or recycling endosomes to lysosomes is a major pathway for LAMP-2A trafficking. PMID- 19136977 TI - Perspective: sex matters: gonadal steroids and the brain. AB - Sex matters to every cell of the body and thus gonadal steroids have the capability of affecting numerous behaviors and neurotransmitters. Several reports in this issue highlight the importance of gonadal steroids: estradiol and progesterone modulate the serotonin transporter in rodents changes in D2 receptors across the menstrual cycle in female cynomolgus monkeys testosterone influences amygdala reactivity in women and temporary suppression of gonadal steroids influences sexual in both men and women. This body of work challenges all of neuropsychopharmacology to consider the importance of sex and gonadal steroids in future studies. PMID- 19136978 TI - The 20 most significant advances in obesity research, prevention and treatment. Foreword. PMID- 19136980 TI - WHO recognition of the global obesity epidemic. AB - The recognition of obesity as a disease was in theory established in 1948 by WHO's (World Health Organization) taking on the International Classification of Diseases but the early highlighting of the potential public health problem in the United States and the United Kingdom 35 years ago was considered irrelevant elsewhere. The medical profession disregarded obesity as important despite the new evidence and WHO data set out in the 1980s. Only in 1995 did WHO find greater problems of overweight than underweight in many developing countries but it required the first special obesity consultation in 1997 and particularly the Millennium burden of disease analyses to suddenly highlight its crucial role in the current unmanageable and escalating medical costs globally. Governments now recognize the overwhelming industrial developments that guarantee an escalating epidemic but neither they nor WHO know how to engage in changing the societal framework to promote routine spontaneous physical activity and a transformation of the food system so that low energy-density food of high nutrient quality becomes the norm. PMID- 19136979 TI - Energy metabolism, fuel selection and body weight regulation. AB - Energy homeostasis is critical for the survival of species. Therefore, multiple and complex mechanisms have evolved to regulate energy intake and expenditure to maintain body weight. For weight maintenance, not only does energy intake have to match energy expenditure, but also macronutrient intake must balance macronutrient oxidation. However, this equilibrium seems to be particularly difficult to achieve in individuals with low fat oxidation, low energy expenditure, low sympathetic activity or low levels of spontaneous physical activity, as in addition to excess energy intake, all of these factors explain the tendency of some people to gain weight. Additionally, large variability in weight change is observed when energy surplus is imposed experimentally or spontaneously. Clearly, the data suggest a strong genetic influence on body weight regulation implying a normal physiology in an 'obesogenic' environment. In this study, we also review evidence that carbohydrate balance may represent the potential signal that regulates energy homeostasis by impacting energy intake and body weight. Because of the small storage capacity for carbohydrate and its importance for metabolism in many tissues and organs, carbohydrate balance must be maintained at a given level. This drive for balance may in turn cause increased energy intake when consuming a diet high in fat and low in carbohydrate. If sustained over time, such an increase in energy intake cannot be detected by available methods, but may cause meaningful increases in body weight. The concept of metabolic flexibility and its impact on body weight regulation is also presented. PMID- 19136981 TI - Fructose: should we worry? AB - Obesity is a growing problem. In the broadest strokes, it is due to a small positive energy balance that persists over a sufficiently long time. Some forms of obesity develop independent of the type of diet that is eaten, whereas others are dependent on the diet. Among the former are individuals with leptin deficiency or genetic defects in the melanocortin 4 receptor. Most human obesity, however, occurs in the presence of highly palatable foods--fat and calorically sweetened beverages. The increase in obesity in the last 35 years has paralleled the increasing use of high-fructose corn syrup (HFCS), which first appeared just before 1970. Current soft drinks and many other foods are sweetened with this product because it is inexpensive and has useful manufacturing properties. The fructose in HFCS and sugar makes beverages very sweet, and this sweetness may underlie the relation of obesity to soft drink consumption. Fructose consumption has also been related to the metabolic syndrome and to abnormal lipid patterns. This evidence suggests that we should worry about our current level of fructose consumption, which has been increasing steadily for over 200 years and now represents over 10% of the energy intake of some people. PMID- 19136982 TI - Physiological and pathophysiological roles of adiponectin and adiponectin receptors in the integrated regulation of metabolic and cardiovascular diseases. AB - Adiponectin and adiponectin receptors (AdipoRs) have been found to play significant roles in the etiology of obesity-related chronic disease. Their discovery has been a long and complicated path, with many challenges. Developing methods to unravel the molecular secrets has been an informative process in itself. However, with both functional and genetic studies confirming adiponectin as a therapeutic target adipokine, many roles and interactions with certain other biomolecules have been clearly defined. We have found that decreased high molecular weight (HMW) adiponectin plays a crucial and causal role in obesity linked insulin resistance and metabolic syndrome; that AdipoR1 and AdipoR2 serve as the major AdipoRs in vivo; and that AdipoR1 activates the AMP kinase (AMPK) pathway and AdipoR2, the peroxisome proliferator-activated receptor alpha (PPARalpha) pathway in the liver, to increase insulin sensitivity and decrease inflammation. Further conclusions are that decreased adiponectin action and increased monocyte chemoattractant protein-1 (MCP-1) form a vicious adipokine network causing obesity-linked insulin resistance and metabolic syndrome; PPARgamma upregulates HMW adiponectin and PPARalpha upregulates AdipoRs; that dietary osmotin can serve as a naturally occurring adiponectin receptor agonist; and finally, that under starvation conditions, MMW adiponectin activates AMPK in hypothalamus, and promotes food intake, and at the same time HMW adiponectin activates AMPK in peripheral tissues, such as skeletal muscle, and stimulates fatty-acids combustion. Importantly, under pathophysiological conditions, such as obesity and diabetes, only HMW adiponectin was decreased; therefore, strategies to increase only HMW adiponectin may be a logical approach to provide a novel treatment modality for obesity-linked diseases, such as insulin resistance and type 2 diabetes. It is hoped that these data will be helpful in developing treatments to counteract the destructive, expensive and painful effects of obesity. PMID- 19136983 TI - Effects of TV time and other sedentary pursuits. AB - Television (TV) viewing is the dominant recreational pastime at all ages, especially for children and adolescents. Many studies have shown that higher TV viewing hours are associated with higher body mass index (BMI), lower levels of fitness and higher blood cholesterol levels. Although the effect size estimated from observational studies is small (with TV viewing explaining very little of the variance in BMI), the results of intervention studies show large effect sizes. The potential mediators of the effect of higher TV viewing on higher BMI include less time for physical activity, reduced resting metabolic rate (for which there is little supporting evidence) and increased energy intake (from more eating while watching TV and a greater exposure to marketing of energy dense foods). Electronic games may have an effect on unhealthy weight gain, but are less related to increased energy intake and their usage is relatively new, making effect size difficult to determine. Thus, TV viewing does not explain much of the differences in body size between individuals or the rise in obesity over time, perhaps because of the uniformly high, but relatively stable, TV viewing hours. Reducing TV viewing hours is a difficult prospect because potential actions, such as social marketing and education, are likely to be relatively weak interventions, although the evidence would suggest that, if viewing could be reduced, it could have a significant impact on reducing obesity prevalence. Regulations to reduce the heavy marketing of energy dense foods and beverages on TV may be the most effective public health measure available to minimize the impact of TV viewing on unhealthy weight gain. PMID- 19136984 TI - Obesity and the built environment: changes in environmental cues cause energy imbalances. AB - The past 30 years have seen dramatic changes in the food and physical activity environments, both of which contribute to the changes in human behavior that could explain obesity. This paper reviews documented changes in the food environment, changes in the physical activity environment and the mechanisms through which people respond to these environments, often without conscious awareness or control. The most important environmental changes have been increases in food accessibility, food salience and decreases in the cost of food. The increases in food marketing and advertising create food cues that artificially stimulate people to feel hungry. The existence of a metabolic pathway that allows excess energy to be stored as fat suggests that people were designed to overeat. Many internal mechanisms favor neurophysiologic responses to food cues that result in overconsumption. External cues, such as food abundance, food variety and food novelty, cause people to override internal signals of satiety. Other factors, such as conditioning and priming, tie food to other desirable outcomes, and thus increase the frequency that hunger is stimulated by environmental cues. People's natural response to the environmental cues are colored by framing, and judgments are flawed and biased depending on how information is presented. People lack insight into how the food environment affects them, and subsequently are unable to change the factors that are responsible for excessive energy consumption. Understanding the causal pathway for overconsumption will be necessary to interrupt the mechanisms that lead to obesity. PMID- 19136985 TI - Obesity research in the next decade. AB - The obesity epidemic demands more insight into genetic predisposition, mechanisms, prevention and therapeutic interventions. Opinions about how to prioritize obesity research in the next decade are many and highly varied. However, in this article I have chosen three areas of focus that arguably should be at the top of the list. These include: (1) the physiologic basis of body weight and body fat regulation; (2) epigenetic mechanisms of energy balance; and (3) the prevention of obesity. The approach needs to be translational and bi directional with a strong emphasis on basic science including studies of relevant gene expression and animal models of energy balance. Clinical research into mechanisms can challenge the existing paradigms that could direct research back to more basic understanding or to applications to populations at risk. Communication between scientists and physicians at the far end of the spectrum needs new and ongoing emphasis. PMID- 19136986 TI - New ligands for melanocortin receptors. AB - Named originally for their effects on peripheral end organs, the melanocortin system controls a diverse set of physiological processes through a series of five G-protein-coupled receptors and several sets of small peptide ligands. The central melanocortin system plays an essential role in homeostatic regulation of body weight, in which two alternative ligands, alpha-melanocyte-stimulating hormone and agouti-related protein, stimulate and inhibit receptor signaling in several key brain regions that ultimately affect food intake and energy expenditure. Much of what we know about the relationship between central melanocortin signaling and body weight regulation stems from genetic studies. Comparative genomic studies indicate that melanocortin receptors used for controlling pigmentation and body weight regulation existed more than 500 million years ago in primitive vertebrates, but that fine-grained control of melanocortin receptors through neuropeptides and endogenous antagonists developed more recently. Recent studies based on dog coat-color genetics revealed a new class of melanocortin ligands, the beta-defensins, which reveal the potential for cross talk between the melanocortin and the immune systems. PMID- 19136987 TI - How much progress have we made over the last few decades? AB - To recognize the 20th anniversary of the opening of the Pennington Biomedical Research Center in Baton Rouge, Louisiana, USA, a conference was held featuring some of the most prominent obesity researchers worldwide, addressing the most significant advances in obesity research. The Center had been founded with a gift from wealthy oilman Claude B Pennington with the mission of 'promoting healthier lives through research and education in nutrition and preventive medicine.' The purpose of the conference was to provide a cohesive presentation of the key events and steps that led to the major discoveries or advances in obesity science and give insight into new research directions. Only 20 of many possible topics could be adequately covered in the program of the conference. Their increasing importance is reflected in the increasing number of related publications as extracted from PubMed. The broad general areas in which these topics fall include measurement and evaluation of obesity, measurement of body composition and fat distribution, adipose tissue biology, genetics, bariatric surgery and public policy. Advances in the areas covered at the conference have had a major impact on our understanding of adipogenesis, central and peripheral regulation of body weight, health consequences of excess body weight, challenges of weight loss, magnitude of the worldwide epidemic and impact of public health policies. PMID- 19136988 TI - Gastrointestinal satiety signals. AB - Obesity constitutes a major global healthcare challenge. The morbidity, mortality, and socioeconomic costs of obesity are considerable. No currently available medical therapy delivers substantial, sustainable weight loss. The need to better understand the mechanisms of appetite regulation is therefore clear. Over the last 20 years, peptide hormones released from the gastrointestinal tract in response to nutritional stimuli have come to be recognized as important physiological regulators of appetite. Hormones such as peptide YY, pancreatic polypeptide, glucagon-like peptide-1 and oxyntomodulin are thought to act as postprandial satiety signals. These physiological pathways of appetite control offer a promising basis for anti-obesity therapies. Here, we briefly review the state of current knowledge of these hormones' actions on brain appetite circuits, and prospects for future research and development. PMID- 19136989 TI - UCP1: its involvement and utility in obesity. AB - Energy balance to prevent the development of obesity is dependent on energy expenditure. Although physical activity is the dominant mechanism for dissipating excess energy, a system of thermogenesis that evolved to protect the body from hypothermia is based upon the uncoupling of oxidative phosphorylation in brown adipocytes by the mitochondrial uncoupling protein (UCP1). It has been shown that upregulation of UCP1 by genetic manipulations or pharmacological agents can reduce obesity and improve insulin sensitivity. Recent evidence has shown the existence of two sources for brown adipocytes, one appearing as discrete brown fat depots during fetal development and the other appears during post-natal development as diffuse populations in traditional white fat depots. The latter can be induced by adrenergic stimulation depending on the genetic background of the animals and the nutritional environment. Understanding the biological and environmental factors controlling the expression of these two brown adipocyte populations promises to provide new strategies by which enhanced thermogenesis can be used to reduce obesity.International Journal of Obesity (2008) 32, S32 S38; doi:10.1038/ijo.2008.236. PMID- 19136990 TI - Advances in adipose tissue metabolism. AB - This review will focus on the recent findings in adipose tissue metabolism with special attention to human adipocyte biology and physiology. There are major advances stemming from the concomitant results obtained from studies on mature human adipocytes, human preadipocytes differentiated in vitro and murine adipose cell lines. Physiological developments have been based on the expanded utilization of various kinds of murine transgenic models and physiological techniques such as microdialysis, open-flow microperfusion, arteriovenous techniques and the utilization of deuterium- or tritium-labelled metabolites that have provided a number of physiological advances in the understanding of human adipose tissue physiology. Gene expression profiling studies and nutrigenomics are emerging methods that herald interesting approaches for the future. An overview of recent discoveries in the mechanisms involved in the control of free fatty acid uptake, triacylglycerol synthesis and fat deposition will be discussed, as well as recent advances in the mechanisms involved in the lipolytic pathways, the role of lipases and perilipins. In addition, the in vivo validation of catecholamine action and the discovery of the lipolytic effects of natriuretic peptides will also be covered. PMID- 19136991 TI - Inflammation and endoplasmic reticulum stress in obesity and diabetes. AB - Obesity is associated with chronic low-grade inflammation. Inflammatory signals interfere with insulin action and disrupt metabolic homeostasis. The c-Jun N terminal kinase (JNK) has been identified as a central mediator of insulin resistance. Recent studies showed that in obesity compromising endoplasmic reticulum (ER) function results in insulin resistance and type 2 diabetes that are dependent on JNK activation. In contrast, enhancing ER function in transgenic mice or by the use of chemical chaperones protects against diet-induced insulin resistance. Hence, ER stress and the related signaling networks present a critical mechanism underlying obesity-induced JNK activity, inflammatory response and insulin resistance. PMID- 19136992 TI - Human obesity as a heritable disorder of the central control of energy balance. AB - In the spirit of celebration associated with the 20th anniversary of the Pennington Biomedical Research Center, we have seized the opportunity of taking a highly personal and not at all comprehensive 'whistle-stop tour' of a large body of evidence that, we feel, supports the following conclusions: (1) that body fat stores are regulated by biological control processes in humans as they are in lower animals; (2) that there are major inherited influences on the efficiency whereby such control processes operate in humans; (3) that the precise nature of those genetic and biological influences and how they interact with environmental factors are beginning to be understood; (4) that most of the genes discovered thus far have their principal impact on hunger, satiety and food intake; (5) that while there is understandable resistance to the notion that genes can influence a human behavior such as the habitual ingestion of food, the implications of these discoveries are essentially benign. Indeed, we hope that they may eventually lead to improved treatment for patients and, in addition, help to inculcate a more enlightened attitude to the obese with a reduction in their experience of social and economic discrimination. PMID- 19136993 TI - Developmental and epigenetic pathways to obesity: an evolutionary-developmental perspective. AB - Although variation in individual lifestyle and genotype are important factors in explaining individual variation in the risk of developing obesity in an obesogenic environment, there is growing evidence that developmentally plastic processes also contribute. These effects are mediated at least in part through epigenetic processes. These developmental pathways do not directly cause obesity but rather alter the risk of an individual developing obesity later in life. At least two classes of developmental pathway are involved. The mismatch pathway involves the evolved adaptive responses of the developing organism to anticipated future adverse environments, which have maladaptive consequences if the environment is mismatched to that predicted. This pathway can be cued by prenatal undernutrition or stresses that lead the organism to forecast an adverse future environment and change its developmental trajectory accordingly. As a result, individuals develop with central and peripheral changes that increase their sensitivity to an obesogenic environment. It provides a model for how obesity emerges in populations in rapid transition, but also operates in developed countries. There is growing experimental evidence that this pathway can be manipulated by, for example, postnatal leptin exposure. Secondly, maternal diabetes, maternal obesity and infant overfeeding are associated with a greater risk of later obesity. Early life offers a potential point for preventative intervention. PMID- 19136994 TI - Insights into energy balance from doubly labeled water. AB - Obesity is defined as the excess storage of energy in the form of fat that results from imbalances between energy intake and expenditure. The study of the components of energy balance has undergone a significant advancement with the application of the doubly labeled water (DLW) method to the measurement of human energy expenditure. This manuscript includes a selective review of the studies that have utilized the doubly labeled method as it applies to the study of human obesity. Although generally now accepted, one of the major surprises from the early applications of DLW was that obese individuals have higher energy expenditures than lean controls. Moreover, weight gain, even in the already obese, is associated with an increase in energy expenditure as weight is one of the strongest predictors of total energy expenditure. Similarly, studies of weight loss treatment show a decrease in energy expenditure due to weight loss and due to adaptive changes in energetic efficiency, but these changes do not account for the common cessation of weight loss observed after 12-26 weeks of restriction. The accumulating data from the application of the DLW method suggest a need to place greater emphasis on mechanisms that lead to a mismatch between energy intake and expenditure rather than a continuing emphasis on energy intake or energy expenditure alone. PMID- 19136995 TI - Development of imaging methods to assess adiposity and metabolism. AB - Body composition studies were first recorded around the time of the renaissance, and advances by the mid-twentieth century facilitated growth in the study of physiology, metabolism and pathological states. The field developed during this early period around the 'two-compartment' molecular level model that partitions body weight into fat and fat-free mass. Limited use was also made of X-rays as a means of estimating fat-layer thickness, but the revolutionary advance was brought about by the introduction of three-dimensional images provided by computed tomography (CT) in the mid 1970s, followed soon thereafter by magnetic resonance imaging (MRI). Complete in vivo reconstruction of all major anatomic body compartments and tissues became possible, thus providing major new research opportunities. This imaging revolution has continued to advance with further methodology refinements including functional MRI, diffusion tensor imaging and combined methods such as positron emission tomography+CT or MRI. The scientific advances made possible by these new and innovative methods continue to unfold today and hold enormous promise for the future of obesity research. PMID- 19136997 TI - The role of fat topology in the risk of disease. AB - Clustering of multiple risk factors such as impaired glucose metabolism, lipid disorders and hypertension has been shown to be the major background of atherosclerotic diseases, and disease entities such as the metabolic syndrome represent a highly atherogenic state. Although these common risks may generally co-exist by accident in one individual, clustering of multiple risk factors in the metabolic syndrome does not occur by accident, and there should be a key player for the syndrome. In 1983, we reported the method for fat analysis using computed tomography scan, which enables us to analyze intra-abdominal visceral adiposity as well as subcutaneous fat. Visceral fat accumulation has been shown to cause impaired glucose metabolism, lipid disorders, and hypertension, and therefore it is considered to be a key player in the metabolic syndrome. To clarify the mechanism by which visceral fat accumulation causes a variety of metabolic and vascular diseases, we studied the molecular characteristics of adipose tissue and adipocytes by investigating expressed genes in visceral and subcutaneous adipocytes and revealed that adipocytes, especially visceral adipocytes, secrete a variety of bioactive substances, the so-called adipocytokines. We showed that visceral fat accumulation causes abnormalities in adipocytokine secretion, such as hypersecretion of plasminogen activator inhibitor 1, which is related to thrombogenic vascular diseases. More importantly, we discovered an important benign adipocytokine named adiponectin, which protects against the development of diabetes mellitus, hypertension, inflammation, and atherosclerotic vascular diseases. Plasma levels of adiponectin decreased in individuals with visceral fat accumulation, and hypoadiponectinemia caused by visceral fat accumulation might be one of the major causes of metabolic syndrome. PMID- 19136996 TI - Leptin receptor signaling and the regulation of mammalian physiology. AB - The adipocyte-derived hormone, leptin, signals the status of body energy stores to the central nervous system to regulate appetite and energy expenditure. A specific long-form leptin receptor (LepRb), a type I cytokine receptor, mediates leptin action on LepRb-expressing neurons in the brain. Leptin binding to LepRb activates the associated Janus kinase-2 (Jak2) tyrosine kinase to promote the phosphorylation of Jak2 and three residues on LepRb; each of these sites mediates a distinct aspect of downstream LepRb signaling, with differing physiologic functions. Tyr(1138) --> STAT3 signaling suppresses feeding, but is not required for a number of other leptin actions. Tyr(985) binds SH2-containing tyrosine phosphatase-2 and suppressor of cytokine signaling-3 and primarily mediates the attenuation of LepRb signaling in vivo. The role for Tyr(1077), the major regulator of signal transducer and activator of transcription-5 (STAT5) during leptin signaling, in the physiologic response to leptin remains unclear, although the obese phenotype of animals deleted for STAT5 in the brain suggests the potential importance of this signaling pathway. Leptin also modulates a number of other signaling pathways in the brain, including PI 3-kinase, mammalian target of rapamycin and AMP-dependent protein kinase; the pathways by which leptin controls these signals remain unclear, however, and may involve some indirect mechanisms. Important issues regarding leptin action and LepRb signaling in the future include not only the more thorough analysis of intracellular signaling pathways, but the neural substrate by which leptin acts, as most major populations of LepRb neurons remain poorly studied. PMID- 19136998 TI - Bariatric surgery and reduction in morbidity and mortality: experiences from the SOS study. AB - Obesity is associated with increased morbidity and mortality. Intentional weight loss results in improvement of cardiovascular risk factors, but most observational studies suggest that weight reduction is associated with increased overall and cardiovascular mortality. No prospective intervention studies on mortality have earlier been reported in obese subjects. The prospective, controlled Swedish Obese Subjects Study enrolled obese subjects who either underwent bariatric surgery (n=2010) or were allocated to a contemporaneously matched, conventionally treated obese control group (n=2037). This review sums up effects on morbidity and mortality over an average of 10 years. The mean weight change of the control group was less than +/-2% over up to 15 years of weight recording. Maximum weight losses in the surgical subgroups were observed after 1 2 years. After 10 years, the weight losses from baseline were stabilized at 25, 16 and 14%, respectively. Bariatric surgery improved all traditional cardiovascular risk states except hypercholesterolemia over 10 years. There were 129 deaths in the control group compared with 101 in the surgery group. The unadjusted overall mortality was reduced by 23.7% (P=0.0419) in the surgery group (relative to controls), whereas the gender-, age- and risk factor-adjusted mortality reduction was 30.7% (P=0.0102). The most common causes of death were myocardial infarction (controls n=25, surgery n=13) and cancer (47/29). Bariatric surgery for severe obesity is associated with long-term weight loss, improved risk factors and decreased overall mortality. PMID- 19137000 TI - Human ribonuclease 9, a member of ribonuclease A superfamily, specifically expressed in epididymis, is a novel sperm-binding protein. AB - To explore the functions of human ribonuclease 9 (RNase 9), we constructed a mammalian fusion expression vector pcDNA-hRNase9, prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences. According to the determined mature protein, recombinant human RNase 9 was prepared in E. coli. Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected, and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay. The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA, but exhibited antibacterial activity, in a concentration/time dependent manner, against E. coli. Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis, but not present in other tissues examined, and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa. These results suggest that human RNase 9 may play roles in host defense of male reproductive tract. PMID- 19136999 TI - Molecular physiology of weight regulation in mice and humans. AB - Evolutionary considerations relating to efficiency in reproduction, and survival in hostile environments, suggest that body energy stores are sensed and actively regulated, with stronger physiological and behavioral responses to loss than gain of stored energy. Many physiological studies support this inference, and suggest that a critical axis runs between body fat and the hypothalamus. The molecular cloning of leptin and its receptor-projects based explicitly on the search for elements in this axis-confirmed the existence of this axis and provided important tools with which to understand its molecular physiology. Demonstration of the importance of this soma-brain reciprocal connection in body weight regulation in humans has been pursued using both classical genetic approaches and studies of physiological responses to experimental weight perturbation. This paper reviews the history of the rationale and methodology of the cloning of leptin (Lep) and the leptin receptor (Lepr), and describes some of the clinical investigation characterizing this axis. PMID- 19137001 TI - Ectopic porcine spermatogenesis in murine subcutis: tissue grafting versus cell injection methods. AB - Fragments of testis tissue from immature animals grow and develop spermatogenesis when grafted onto subcutaneous areas of immunodeficient mice. The same results are obtained when dissociated cells from immature testes of rodents are injected into the subcutis of nude mice. Those cells reconstitute seminiferous tubules and facilitate spermatogenesis. We compared these two methods, tissue grafting and cell-injection methods, in terms of the efficiency of spermatogenesis in the backs of three strains of immunodeficient mice, using neonatal porcine testicular tissues and cells as donor material. Nude, severe combined immunodeficient (SCID) and NOD/Shi-SCID, IL-2Rgammacnull (NOG) mice were used as recipients. At 10 months after surgery, the transplants were examined histologically. Both grafting and cell-injection methods resulted in porcine spermatogenesis on the backs of recipient mice; the percentage of spermatids present in the transplants was 67% and 22%, respectively. Using the grafting method, all three strains of mice supported the same extent of spermatogenesis. As for the cell-injection method, although SCID mice were the best host for supporting reconstitution and spermatogenesis, any difference from the other strains was not significant. As NOG mice did not show any better results, the severity of immunodeficiency seemed to be irrelevant for supporting xeno-ectopic spermatogenesis. Our results confirmed that tubular reconstitution is applicable to porcine testicular cells. This method as well as the grafting method would be useful for studying spermatogenesis in different kinds of animals. PMID- 19137002 TI - Androgen receptor gene polymorphism and sex hormones in elderly men: the Tromso study. AB - The aim of this study was to examine whether CAG/GGN repeats are significant modulators of serum concentrations of total and free testosterone (T) as well as of luteinizing hormone (LH) in elderly men. Sixty-nine 60- to 80-year-old men with subnormal T levels (< or = 11.0 nmol L(-1)) and 104 men with normal T levels taking part in a nested case-control study were used for these analyses. Sex hormones were measured and free T was calculated. The CAG and GGN polymorphisms in the androgen receptor gene were determined by polymerase chain reaction and subsequent direct sequencing. There were no differences in the CAG and GGN repeat lengths between the groups. In cross-sectional analyses of the whole cohort, total and free T were positively associated with CAG length (all P < 0.05) before, but not after, waist circumference or body mass index was added to the model. CAG repeat lengths were weakly, but not independently, associated with total and free T. These findings indicate that when clinically evaluating T and LH levels in elderly men, the CAG and GGN repeat lengths do not need to be taken into consideration. PMID- 19137003 TI - Pediatric Highlights' fifth anniversary in the International Journal of Obesity. PMID- 19137005 TI - miRNA in situ hybridization in formaldehyde and EDC-fixed tissues. AB - MicroRNAs are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in substantial microRNA loss from mouse tissue sections, which can be prevented by fixation with 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide that irreversibly immobilizes the microRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid probes by recording nucleic acid melting temperature during ISH. PMID- 19137006 TI - Large-scale profiling of protein palmitoylation in mammalian cells. AB - S-palmitoylation is a pervasive post-translational modification required for the trafficking, compartmentalization and membrane tethering of many proteins. We demonstrate that the commercially available compound 17-octadecynoic acid (17 ODYA) can serve as a bioorthogonal, click chemistry probe for in situ labeling, identification and verification of palmitoylated proteins in human cells. We identified approximately 125 predicted palmitoylated proteins, including G proteins, receptors and a family of uncharacterized hydrolases whose plasma membrane localization depends on palmitoylation. PMID- 19137007 TI - Role of miR-143 targeting KRAS in colorectal tumorigenesis. AB - Dysregulated expression of microRNAs (miRNAs) is associated with a variety of diseases, including colorectal cancer. By comparing more than 200 miRNAs in 13 pairs of matched colorectal cancer and normal adjacent tissue samples through qRT PCR and microarray analysis, we found a widespread disruption of miRNA expression during colorectal tumorigenesis. In particular, among a panel of presumed targets generated by in silico analysis that may interact with these aberrantly expressed miRNAs, KRAS oncogene has been further experimentally validated as the target of miR-143. First, an inverse correlation between KRAS protein and miR-143 in vivo was found. Second, KRAS expression in Lovo cells was significantly abolished by treatment with miR-143 mimic, whereas miR-143 inhibitor increased KRAS protein level. Third, luciferase reporter assay confirmed that miR-143 directly recognize the 3'-untranslated region of KRAS transcripts. Four, Lovo cells treated with miR 143 inhibitor showed a stimulated cell proliferation, whereas miR-143 overexpression had an opposite effect. Finally, inhibition of KRAS expression by miR-143 inhibits constitutive phosphorylation of ERK1/2. Taken together, the present study provides the first evidences that miR-143 is significant in suppressing colorectal cancer cell growth through inhibition of KRAS translation. PMID- 19137008 TI - Sprouty4 negatively regulates protein kinase C activation by inhibiting phosphatidylinositol 4,5-biphosphate hydrolysis. AB - Sproutys have been shown to negatively regulate growth factor-induced extracellular signal-regulated kinase (ERK) activation, and suggested to be an anti-oncogene. However, molecular mechanism of the suppression has not yet been clarified completely. Sprouty4 inhibits vascular endothelial growth factor (VEGF) A-induced ERK activation, but not VEGF-C-induced ERK activation. It has been shown that VEGF-A-mediated ERK activation is strongly dependent on protein kinase C (PKC), whereas that by VEGF-C is dependent on Ras. This suggests that Sprouty4 inhibits the PKC pathway more specifically than the Ras pathway. In this study, we confirmed that Sprouty4 suppressed various signals downstream of PKC, such as phosphorylation of MARCKS and protein kinase D (PKD), as well as PKC-dependent nuclear factor (NF)-kappaB activation. Furthermore, Sprouty4 suppressed upstream signals of PKC, such as Ca(2+) mobilization, phosphatidylinositol 4,5-biphosphate (PIP(2)) breakdown and inositol 1,4,5-triphosphate (IP(3)) production in response to VEGF-A. Those effects were dependent on the C-terminal cysteine-rich region, but not on the N-terminal region of Sprouty4, which is critical for the suppression of fibroblast growth factor (FGF)-mediated ERK activation. Sprouty4 overexpression or deletion of the Sprouty4 gene did not affect phospholipase C (PLC) gamma-1 activation, which is an enzyme that catalyzes PIP(2) hydrolysis. Moreover, Sprouty4 inhibited not only VEGF-A-mediated PIP(2) hydrolysis but also inhibited the lysophosphatidic acid (LPA)-induced PIP(2) breakdown that is catalyzed by PLC beta/epsilon activated by G-protein coupled receptor (GPCR). Taken together, Sprouty4 has broader suppression activity for various stimuli than previously thought; it may function as an inhibitor for various types of PLC dependent signaling as well as for ERK activation. PMID- 19137009 TI - Activation of the non-canonical Dvl-Rac1-JNK pathway by Frizzled homologue 10 in human synovial sarcoma. AB - We previously reported that Frizzled homologue 10 (FZD10), a member of the Wnt signal receptor family, was highly and specifically upregulated in synovial sarcoma and played critical roles in its cell survival and growth. We here report a possible molecular mechanism of the FZD10 signaling in synovial sarcoma cells. We found a significant enhancement of phosphorylation of the Dishevelled (Dvl)2/Dvl3 complex as well as activation of the Rac1-JNK cascade in synovial sarcoma cells in which FZD10 was overexpressed. Activation of the FZD10-Dvls-Rac1 pathway induced lamellipodia formation and enhanced anchorage-independent cell growth cells. FZD10 overexpression also caused the destruction of the actin cytoskeleton structure, probably through the downregulation of the RhoA activity. Our results have strongly implied that FZD10 transactivation causes the activation of the non-canonical Dvl-Rac1-JNK pathway and plays critical roles in the development/progression of synovial sarcomas. PMID- 19137010 TI - Ceramide synthase 6 modulates TRAIL sensitivity and nuclear translocation of active caspase-3 in colon cancer cells. AB - We have previously shown that the death receptor ligand TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) induces an increase of intracellular C(16)-ceramide in sensitive SW480 but not in resistant SW620 cells. Resistance in SW620 cells was overcome by exogenous ceramide, leading us to propose that defective ceramide signaling contributes to TRAIL resistance. In this study we found that the increase in C(16)-ceramide in SW480 cells was inhibited by fumonisin B1, an inhibitor of ceramide synthases (CerS). Protein analysis revealed that TRAIL-resistant SW620 cells expressed lower levels of ceramide synthase 6 (CerS6, also known as longevity assurance homologue 6), which prompted us to investigate the effect of CerS6 modulation on TRAIL phenotype. RNAi against CerS6 resulted in a specific and significant decrease of the C(16)-ceramide species, which was sufficient to inhibit TRAIL-induced apoptosis. In cells with decreased levels of CerS6, caspase-3 was activated but failed to translocate into the nucleus. CerS6 localized primarily to the perinuclear region, suggesting this enzyme may be important in regulation of nuclear permeability. Moderate elevation in CerS6 expression was sufficient to reverse TRAIL resistance in SW620 cells. These results suggest that modulation of CerS6 expression may constitute a new therapeutic strategy to alter apoptotic susceptibility. PMID- 19137011 TI - The STAT3 inhibitor NSC 74859 is effective in hepatocellular cancers with disrupted TGF-beta signaling. AB - Hepatocellular carcinoma (HCC) is the third leading cause of cancer deaths worldwide, with few effective therapeutic options for advanced disease. At least 40% of HCCs are clonal, potentially arising from STAT3+, NANOG+ and OCT3/4+ liver progenitor/stem cell transformation, along with inactivation of transforming growth factor-beta (TGF-beta) signaling. Here we report significantly greater signal transducer and activator of transcription 3 (STAT3) and tyrosine phosphorylated STAT3 in human HCC tissues (P<0.0030 and P<0.0455, respectively) than in human normal liver. Further, in HCC cells with loss of response to TGF beta, NSC 74859, a STAT3-specific inhibitor, markedly suppresses growth. In contrast, CD133(+) status did not affect the response to STAT3 inhibition: both CD133(+) Huh-7 cells and CD133(-) Huh-7 cells are equally sensitive to NSC 74859 treatment and STAT3 inhibition, with an IC(50) of 100 muM. Thus, the TGF beta/beta2 spectrin (beta2SP) pathway may reflect a more functional 'stem/progenitor' state than CD133. Furthermore, NSC 74859 treatment of Huh-7 xenografts in nude mice significantly retarded tumor growth, with an effective dose of only 5 mg/kg. Moreover, NSC 74859 inhibited tyrosine phosphorylation of STAT3 in HCC cells in vivo. We conclude that inhibiting interleukin 6 (IL6)/STAT3 in HCCs with inactivation of the TGF-beta/beta2SP pathway is an effective approach in management of HCCs. Thus, IL6/STAT3, a major signaling pathway in HCC stem cell renewal and proliferation, can provide a novel approach to the treatment of specific HCCs. PMID- 19137012 TI - MYC-induced myeloid leukemogenesis is accelerated by all six members of the antiapoptotic BCL family. AB - Signals that control the fine balance between cell death and cell survival are altered during tumorigenesis. Understanding the mechanisms by which this balance is perturbed, leading to excessive cell survival, is important for designing effective therapies. Proteins belonging to the B-cell lymphoma (BCL) family are known to regulate death responses to apoptotic signals, especially those originating within cells. A subset of BCL family members capable of inhibiting cell death is known to contribute to tumorigenesis; however, it is not known whether all six antiapoptotic BCL family members play a causal role in tumor development. Using a mouse model of MYC-driven leukemia, we showed that, in addition to the well characterized BCL2 and BCLxl (BCL2L1), the other four family members -- BCLw (BCL2L2), BCLb (BCL2L10), BFL1 (BCL2A1) and MCL1 -- also cooperate with MYC to accelerate leukemogenesis. In addition, high levels of each family member are found in either solid human tumors or cell lines derived from human leukemias or lymphomas. PMID- 19137013 TI - EGR-1 forms a complex with YAP-1 and upregulates Bax expression in irradiated prostate carcinoma cells. AB - In this study, we investigated the functional role of early growth response-1 (Egr1 gene) in the regulation of radiation-induced clonogenic inhibition and apoptosis in p53 wild-type and mutant prostate cancer cells 22Rv1 and DU145, respectively. 22Rv1 cells were more sensitive to irradiation compared with DU145 cells, and the sensitivity was enhanced by overexpression of EGR-1 in both cells. Dominant-negative EGR-1 mutant (dnEGR-1) or repressor of EGR-1, NGFIA binding protein 1 (NAB1), increased radioresistance of these cells. Significant activation of caspases 3 and 9 and Bcl2-associated X (Bax) with increased poly(ADP-ribose) polymerase (PARP) cleavage and cytochrome c release was observed in radiation-exposed EGR-1 overexpressing cells. Gel shift analysis and chloramphenicol acetyl transferase (CAT) reporter assays indicate that EGR-1 transactivates the promoter of the Bax gene. Interaction of EGR-1 and Yes kinase associated protein 1 (YAP-1) through the WW domain of YAP-1 enhances the transcriptional activity of EGR-1 on the Bax promoter as shown by chromatin immunoprecipitation and reporter assays. Irradiation of PC3 cell xenografts that were treated with adenoviral EGR-1 showed significant regression in tumor volume. These findings establish the radiation-induced pro-apoptotic action of EGR-1, in a p53-independent manner, by directly transactivating Bax, and prove that alters the B-cell CLL/lymphoma 2 (Bcl-2)/Bax ratio as one of the mechanisms resulting in significant activation of caspases, leading to cell death through the novel interaction of EGR-1 with YAP-1. PMID- 19137014 TI - Mouse embryonic fibroblasts null for the Kruppel-like factor 4 gene are genetically unstable. AB - Kruppel-like factor 4 (KLF4) is a zinc-finger transcription factor with tumor suppressive activity in colorectal cancer. Here, we investigated whether KLF4 is involved in maintaining genetic stability in mouse embryonic fibroblasts (MEFs) isolated from mice wild type (+/+), heterozygous (+/-), or homozygous (-/-) for the Klf4 alleles. Compared to Klf4(+/+) and Klf4(+/-) MEFs, Klf4(-/-) MEFs had both a higher level of apoptosis and rate of proliferation. Quantification of chromosome numbers showed that Klf4(-/-) MEFs were aneuploid. A higher number of Klf4(-/-) MEFs exhibited gamma-H2AX foci and had higher amounts of gamma-H2AX compared to controls. Cytogenetic analysis demonstrated the presence of numerous chromosome aberrations including dicentric chromosomes, chromatid breaks, and double minute chromosomes in Klf4(-/-) cells but in few, if any, Klf4(+/+) or Klf4(+/-) MEFs. Approximately 25% of Klf4(-/-) MEFs exhibited centrosome amplification in contrast to the less than 5% of Klf4(+/+) or Klf4(+/-) MEFs. Finally, only Klf4(-/-) MEFs were capable of anchorage-independent growth. Taken together, these findings demonstrate that MEFs null for the Klf4 alleles are genetically unstable, as evidenced by the presence of aneuploidy, chromosome aberration and centrosome amplification. The results support a crucial role for KLF4 in maintaining genetic stability and as a tumor suppressor. PMID- 19137015 TI - TGF-beta repression of Id2 induces apoptosis in gut epithelial cells. AB - Transforming growth factor-beta (TGF-beta) regulates epithelial tissue homeostasis by activating processes that control cell cycle arrest, differentiation and apoptosis. Disruption of the TGF-beta signaling pathway often occurs in colorectal cancers. Earlier, we have shown that TGF-beta induces apoptosis through the transcription factor Smad3. Affymetrix oligonucleotide microarrays were used to identify TGF-beta/Smad3 target genes that regulate apoptosis in rat intestinal epithelial cells (RIE-1). We found that TGF-beta repressed the expression of the inhibitor of differentiation (Id) gene family. Knockdown of Id1 and Id2 gene expression induced apoptosis in RIE-1 cells, whereas overexpression of Id2 attenuated TGF-beta-induced apoptosis. TranSignal Protein/DNA arrays were used to identify the hypoxia-inducing factor-1 (HIF-1) as a downstream target of TGF-beta. HIF-1 is a basic helix-loop-helix protein, and overexpression of Id2 blocked HIF-1 activation by TGF-beta. Furthermore, knockdown of HIF-1 blocked TGF-beta-induced apoptosis. Thus, we have identified HIF-1 as a novel mediator downstream of Id2 in the pathway of TGF-beta-induced apoptosis. PMID- 19137016 TI - 9-Aminoacridine-based anticancer drugs target the PI3K/AKT/mTOR, NF-kappaB and p53 pathways. AB - Acquisition of a transformed phenotype involves deregulation of several signal transduction pathways contributing to unconstrained cell growth. Understanding the interplay of different cancer-related signaling pathways is important for development of efficacious multitargeted anticancer drugs. The small molecule 9 aminoacridine (9AA) and its derivative, the antimalaria drug quinacrine, have selective toxicity for tumor cells and can simultaneously suppress nuclear factor kappaB (NF-kappaB) and activate p53 signaling. To investigate the mechanism underlying these drug activities, we used a combination of two-dimensional protein separation by gel electrophoresis and mass spectrometry to identify proteins whose expression is altered in tumor cells by 9AA treatment. We found that 9AA treatment results in selective downregulation of a specific catalytic subunit of the phosphoinositide 3-kinase (PI3K) family, p110 gamma. Further exploration of this observation demonstrated that the mechanism of action of 9AA involves inhibition of the prosurvival AKT/mammalian target of rapamycin (mTOR) pathway that lies downstream of PI3K. p110 gamma translation appears to be regulated by mTOR and feeds back to further modulate mTOR and AKT, thereby impacting the p53 and NF-kappaB pathways as well. These results reveal functional interplay among the PI3K/AKT/mTOR, p53 and NF-kappaB pathways that are frequently deregulated in cancer and suggest that their simultaneous targeting by a single small molecule such as 9AA could result in efficacious and selective killing of transformed cells. PMID- 19137017 TI - A novel protein, MAPO1, that functions in apoptosis triggered by O6-methylguanine mispair in DNA. AB - O(6)-Methylguanine produced in DNA induces mutation due to its ambiguous base pairing properties during DNA replication. To suppress such an outcome, organisms possess a mechanism to eliminate cells carrying O(6)-methylguanine by inducing apoptosis that requires the function of mismatch repair proteins. To identify other factors involved in this apoptotic process, we performed retrovirus mediated gene-trap mutagenesis and isolated a mutant that acquired resistance to a simple alkylating agent, N-methyl-N-nitrosourea (MNU). However, it was still sensitive to methyl methanesulfonate, 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3 (2-chloroethyl)-3-nitrosourea, etoposide and ultraviolet irradiation. Moreover, the mutant exhibited an increased mutant frequency after exposure to MNU. The gene responsible was identified and designated Mapo1 (O(6)-methylguanine-induced apoptosis 1). When the expression of the gene was inhibited by small interfering RNA, MNU-induced apoptosis was significantly suppressed. In the Mapo1-defective mutant cells treated with MNU, the mitochondrial membrane depolarization and caspase-3 activation were severely suppressed, although phosphorylation of p53, CHK1 and histone H2AX was observed. The orthologs of the Mapo1 gene are present in various organisms from nematode to humans. Both mouse and human MAPO1 proteins expressed in cells localize in the cytoplasm. We therefore propose that MAPO1 may play a role in the signal-transduction pathway of apoptosis induced by O(6) methylguanine-mispaired lesions. PMID- 19137018 TI - Specific regulation of mRNA cap methylation by the c-Myc and E2F1 transcription factors. AB - Methylation of the mRNA 5' guanosine cap is essential for efficient gene expression. The 5' methyl cap binds to eIF4E, which is the first step in the recruitment of mRNA to the 40S ribosomal subunit. To investigate whether mRNA cap methylation is regulated in a gene-specific manner, we established a method to detect the relative level of cap methylation on specific mRNAs. We found that two transcription factors, c-Myc and E2F1, induce cap methylation of their transcriptional target genes, and therefore, c-Myc and E2F1 upregulate gene expression by simultaneously inducing transcription and promoting translation. c Myc-induced cap methylation is greater than transcriptional induction for the majority of its target genes, indicating that this is a major mechanism by which Myc regulates gene expression. PMID- 19137019 TI - Constitutive activation and targeted disruption of signal transducer and activator of transcription 3 (Stat3) in mouse epidermis reveal its critical role in UVB-induced skin carcinogenesis. AB - In this study, the potential role of Stat3 in UVB-induced skin carcinogenesis was examined using skin-specific gain and loss of function transgenic mice, that is, K5.Stat3C and K5Cre.Stat3(fl/fl) mice, respectively. The epidermis of Stat3 deficient mice was highly sensitive to UVB-induced apoptosis, whereas the epidermis of K5.Stat3C mice was more resistant to UVB-induced apoptosis. In particular, the status of Stat3 influenced the survival of ultraviolet photoproduct cells, including those located in the hair follicles. K5.Stat3C mice exhibited significantly increased epidermal proliferation and hyperplasia in response to UVB irradiation, whereas Stat3-deficient mice showed reduced epidermal proliferation and hyperplasia. Expression of target genes regulated by Stat3, such as cyclin D1 and Bcl-x(L), was increased in epidermis of both control and UVB-irradiated K5.Stat3C mice, and downregulated in epidermis of both control and UVB-irradiated K5Cre.Stat3(fl/fl) mice. Following UVB irradiation, the formation of skin tumors in K5.Stat3C mice was accelerated and both the incidence and multiplicity of skin tumors were significantly greater than wild-type controls. In contrast, Stat3-deficient mice were resistant to UVB skin carcinogenesis. These results show that Stat3 plays an important role in the development of UVB-induced skin tumors through its effects on both survival and proliferation of keratinocytes during carcinogenesis. PMID- 19137020 TI - Canonical WNT signalling determines lineage specificity in Wilms tumour. AB - Wilms tumours (WTs) have two distinct types of histology with or without ectopic mesenchymal elements, suggesting that WTs arise from either the mesenchymal or epithelial nephrogenic lineages. Regardless of the presence or absence of CTNNB1 mutations, nuclear accumulation of beta-catenin is often observed in WTs with ectopic mesenchymal elements. Here, we addressed the relationship between the WNT signalling pathway and lineage in WTs by examining CTNNB1 and WT1 mutations, nuclear accumulation of beta-catenin, tumour histology and gene expression profiles. In addition, we screened for mutations in WTX, which has been proposed to be a negative regulator of the canonical WNT-signalling pathway. Unsupervised clustering analysis identified two classes of tumours: mesenchymal lineage WNT dependent tumours, and epithelial lineage WNT-independent tumours. In contrast to the mesenchymal lineage specificity of CTNNB1 mutations, WTX mutations were surprisingly observed in both lineages. WTX-mutant WTs with ectopic mesenchymal elements had nuclear accumulation of beta-catenin, upregulation of WNT target genes and an association with CTNNB1 mutations in exon 7 or 8. However, epithelial lineage WTs with WTX mutations had no indications of active WNT signalling, suggesting that the involvement of WTX in the WNT-signalling pathway may be lineage dependent, and that WTX may have an alternative function to its role in the canonical WNT-signalling pathway. PMID- 19137021 TI - FEN1 contributes to telomere stability in ALT-positive tumor cells. AB - Abrogation of telomere stability through loss-of-function mutations in telomere binding proteins contributes to genomic instability and cancer progression. Recently, Flap endonuclease 1 (FEN1) was shown to contribute to telomere stability in human cells that had not yet activated a telomere maintenance mechanism, suggesting that abrogation of FEN1 function influences the transformation process by compromising telomere stability and driving genomic instability. Here, we analyse the telomeres in human cancer cells following FEN1 depletion. We show that FEN1 is required for telomere stability in cells that rely on the alternative lengthening of telomere (ALT) mechanism. Indeed, FEN1 depletion resulted in telomere dysfunction, characterized by formation of telomere dysfunction-induced foci (TIFs) and end-to-end fusions in ALT-positive cells. In contrast, no telomere phenotype was observed in telomerase-positive cells on FEN1 depletion, suggesting that ongoing telomerase activity protected telomeres. In consonance with this, we found that expression of the catalytic component of telomerase (hTERT) but not an inactive allele rescued telomere dysfunction on FEN1 depletion in ALT cells. Our data suggest that mutations that arise in FEN1 affect telomere stability and genome fidelity by promoting telomere fusions and anaphase-bridge-breakage cycles, which further drive genome instability and thereby contribute to the transformation process. PMID- 19137023 TI - Microstructure of Mo/Si multilayers with B4C diffusion barrier layers. AB - The growth behavior of B(4)C interlayers deposited at the interfaces of Mo/Si multilayers was investigated using x-ray photoemission spectroscopy, x-ray reflectivity, and x-ray diffraction measurements. We report an asymmetry in the formation of B(4)C at the B(4)C-on-Mo interface compared to the B(4)C-on-Si interface. X-ray photoelectron spectroscopy (XPS) depth profiling shows that for B(4)C-on-Mo the formed stoichiometry is close to expectation (4:1 ratio), while for B(4)C-on-Si it is observed that carbon diffuses from the B(4)C interfaces into the multilayer, resulting in nonstochiometric growth (>4:1). As a result, there is a discrepancy in the optical response near 13.5 nm wavelength, where B(4)C-on-Mo behaves according to model simulations, while B(4)C-on-Si does not. The as-deposited off-stoichiometric B(4)C-on-Si interface also explains why these interfaces show poor barrier properties against temperature induced interdiffusion. We show that the stoichiometry of B(4)C at the Mo-Si interfaces is connected to the structure of the layers onto which B(4)C is grown. Because of enhanced diffusion into the amorphous Si surface, we suggest that deposited boron and carbon atoms form Si(X)B(Y) and Si(X)C(Y) compounds. The low formation enthalpy of Si(X)C(Y) ensures C depletion of any B(X)C(Y) interlayer. Only after a saturated interfacial layer is formed, does further deposition of boron and carbon atoms result in actual B(4)C formation. In contrast to the off stoichiometric B(4)C growth on top of Si, B(4)C grown on top of Mo retains the correct stoichiometry because of the higher formation enthalpies for Mo(X)B(Y) and Mo(X)C(Y) formation and the limited diffusion depth into the (poly) crystalline Mo surface. PMID- 19137022 TI - Role of chromodomain helicase DNA-binding protein 2 in DNA damage response signaling and tumorigenesis. AB - The chromodomain helicase DNA-binding proteins (CHDs) are known to affect transcription through their ability to remodel chromatin and modulate histone deacetylation. In an effort to understand the functional role of the CHD2 in mammals, we have generated a Chd2 mutant mouse model. Remarkably, the Chd2 protein appears to play a critical role in the development, hematopoiesis and tumor suppression. The Chd2 heterozygous mutant mice exhibit increased extramedullary hematopoiesis and susceptibility to lymphomas. At the cellular level, Chd2 mutants are defective in hematopoietic stem cell differentiation, accumulate higher levels of the chromatin-associated DNA damage response mediator, gamma H2AX, and exhibit an aberrant DNA damage response after X-ray irradiation. Our data suggest a direct role for the chromatin remodeling protein in DNA damage signaling and genome stability maintenance. PMID- 19137024 TI - Photorefractive dynamic properties of Ca2+-doped strontium barium niobate crystals. AB - Measurements for photorefractive dynamic properties under polarized light with different luminous intensity and wavelength at 650 nm were performed in calcium doped strontium barium niobate crystals as a function of the time. Experimental results further validated that the different input luminous intensities influence only the saturated time and photorefractive velocity of the photorefractive crystals. Birefringence experiments suggest that an input luminous intensity oscillatory threshold should exist between 64 mW/cm(2) and 13 mW/cm(2). The dynamic change of refractive index from self-focusing to asymmetric self defocusing was also measured and regarded as the mutual results of the nonuniform intensity incidence and positive thermal lens effect, which was validated by a further experiment based on the Michelson interference. Also, oscillation accompanying change in the dynamic indices was analyzed and attributed to the high dielectric effect as well as asymmetric self-defocusing in this investigation. PMID- 19137025 TI - On-axis probability density function and fade behavior of partially coherent beams propagating through turbulence. AB - We examine the gamma-gamma and lognormal distributions as they apply to the irradiance at a point detector produced by partially coherent beams propagating horizontally through atmospheric turbulence. Our investigation compares the probability density functions and probability of fade predicted by the distributions with results from a wave-optics simulation developed for partially coherent beam propagation. For a partially coherent beam that is not too far removed from a coherent beam, we find the wave-optics results tend to the gamma gamma model for the weak fluctuation regime and the results are closer to the lognormal model for the strong fluctuation regime. We observe that increasing the initial beam size/Fried parameter ratio (w(0)/r(0)) or shortening the coherence length (l(c)) tends to narrow the probability density profile produced by the simulation. PMID- 19137026 TI - Quasi-analytical determination of noise-induced error limits in lidar retrieval of aerosol backscatter coefficient by the elastic, two-component algorithm. AB - The elastic, two-component algorithm is the most common inversion method for retrieving the aerosol backscatter coefficient from ground- or space-based backscatter lidar systems. A quasi-analytical formulation of the statistical error associated to the aerosol backscatter coefficient caused by the use of real, noise-corrupted lidar signals in the two-component algorithm is presented. The error expression depends on the signal-to-noise ratio along the inversion path and takes into account "instantaneous" effects, the effect of the signal-to noise ratio at the range where the aerosol backscatter coefficient is being computed, as well as "memory" effects, namely, both the effect of the signal-to noise ratio in the cell where the inversion is started and the cumulative effect of the noise between that cell and the actual cell where the aerosol backscatter coefficient is evaluated. An example is shown to illustrate how the "instantaneous" effect is reduced when averaging the noise-contaminated signal over a number of cells around the range where the inversion is started. PMID- 19137027 TI - Performance characterization of integral imaging systems based on human vision. AB - The perceptual contrast threshold (PCT) surface is proposed for characterizing the systematic performance of integral imaging (InI) systems. The method to determine the PCT surface of InI systems is presented first. The theoretical model of the PCT surface is then derived by considering the integral contribution of an InI system as well as the human visual system. Preliminary simulated results show that the PCT can correctly describe the InI properties related to the lateral resolution, contrast, depth distance, and their trade-off relationships, which can be used to compare the systematic performance of InI systems with different design parameters. To the best of our knowledge, this is the first study of the relationship of the resolvable InI contrast with the lateral resolution and the object depth. PMID- 19137028 TI - Edge technique for direct detection of strain and temperature based on optical time domain reflectometry. AB - A hybrid technique for real-time direct detection of strain and temperature along a single-mode fiber is proposed. The temperature is directly detected from the Raman backscattering in the time domain. To retrieve the strain profile from the Brillouin backscattering, an edge technique is introduced and a response function of the Fabry-Perot interferometer for the Brillouin backscattering is defined for the first time to our knowledge. The outgoing laser and the Brillouin backscattering are measured on different interference orders through different channels of the Fabry-Perot interferometer. A low-resolution reference channel and a high-resolution Brillouin channel are designed to keep both a high measurement sensitivity and a wide dynamic range. The measurement is based on detecting the bandwidth changes and the frequency shifts of the Brillouin backscattering; thus the resulting measurement is insensitive to the power fluctuation of the backscattering and the laser frequency jitter or drift. Neither time-consuming frequency scanning nor heavy data processing is needed, which makes real-time detection possible. The dynamic range of the edge technique can be increased substantially by using a piezoelectric tunable and capacitive servo-stabilized Fabry-Perot interferometer. We highlight the potential of this technique by numerical simulations. Given that the uncertainty of the temperature measurement is 0.5 degrees C and that the spatial and temporal resolutions are 10 cm and 1 s, the strain uncertainty is less than 20 microepsilon within a 2 km distance when the strain is below 0.4%, and it is not more than 110 microepsilon within a 4 km distance when the strain is below 0.6%. PMID- 19137029 TI - Probing technique using circular motion of a microsphere controlled by optical pressure for a nanocoordinate measuring machine. AB - A new surface probing technique using the circular motion of an optically-trapped microsphere is proposed for a nanocoordinate measuring system. The probe sphere is oscillated circularly in the plane perpendicular to the probe axis and the circular orbit of the probe sphere is monitored for the detection of the position and normal vector direction of the surface. The principle of detection is based on changes in the circular orbit of the microsphere. When the probe approaches a work surface, the orbit of the probe sphere becomes elliptical. The minor-axis length and the minor-axis angle of the ellipse are then used as parameters to detect the position and normal vector direction of the surface, respectively. In this study, the circular motion probe is shown to have a resolution of position detection of 39 nm, and the accuracy of measuring a normal vector to the surface is on the order of 3 degrees. PMID- 19137030 TI - Parabolic tapered structure for an ultracompact multimode interference coupler. AB - A parabolic tapered structure based on general interference has been proposed and studied theoretically with silica waveguides of silicon oxinitride (SiON) core by using a mathematical model based on sinusoidal modes for the reduction of coupling length of a 2x2 multimode interference (MMI) coupler. The coupling behaviors of the proposed structure are compared with those of other MMI structures. It is seen that the beat length for the proposed tapered MMI coupler is approximately half of that of a conventional MMI coupler. The effect of power imbalance on the fabrication tolerance of a 3 dB coupler using the proposed tapered structure is also studied and compared with that of other MMI structures. PMID- 19137031 TI - Measurement method for photoluminescent quantum yields of fluorescent organic dyes in polymethyl methacrylate for luminescent solar concentrators. AB - A method for measuring the photoluminescent quantum yields (PLQY) of luminescent organic dyes is presented. The self-absorption probability calculated at different dye concentrations is used to determine the absolute quantum yield from the observed values. The results for a range of commercially available dyes show high quantum yields, even at high concentrations, and an absence of quenching. The PLQY of several dye mixtures are also presented. The results indicate an absence of any reduction of PLQY in a dye mixture as compared with the individual PLQY of the dyes. PMID- 19137032 TI - Fundus camera systems: a comparative analysis. AB - Retinal photography requires the use of a complex optical system, called a fundus camera, capable of illuminating and imaging the retina simultaneously. The patent literature shows two design forms but does not provide the specifics necessary for a thorough analysis of the designs to be performed. We have constructed our own designs based on the patent literature in optical design software and compared them for illumination efficiency, image quality, ability to accommodate for patient refractive error, and manufacturing tolerances, a comparison lacking in the existing literature. PMID- 19137033 TI - On the accuracy of generalized Fokker-Planck transport equations in tissue optics. AB - Forward-peaked and large-angle scattering approximations of the radiative transport equation give rise to generalized Fokker-Planck equations whose main feature is the replacement of the integral scattering operator with differential operators in the direction-space variables. Using the P(N) method, an appraisal of generalized Fokker-Planck equations due to Gonzalez-Rodriguez and Kim [Appl. Opt.47, 2599-2609 (2008)], Leakeas and Larsen [Nucl. Sci. Eng.137, 236-250 (2001), and J. Opt. Soc. Am. A20, 92-98 (2003)], and Pomraning [Math. Models Meth. Appl. Sci.2, 21-36 (1992)] is carried out by computing the relative error between the backscattered and transmitted surface flux predicted by the generalized Fokker-Planck equations and the transport equation with Henyey Greenstein phase function for anisotropies ranging from 0 to 1. Generalized Fokker-Planck equations whose scattering operators incorporate large-angle scattering and possess eigenvalues similar to the integral scattering operator with Henyey-Greenstein phase function are found to minimize the relative error in the limit of unit anisotropy. PMID- 19137034 TI - Approach to investigating the spherical aberration variation with the object position in an aplanatic system. AB - In a general optical system, spherical aberration will arise when the on-axis position of an object is changed from its optimum position to another point. Induced spherical aberration can be used to compensate the aberration caused by inserting or removing a medium plate with any thickness that has a refractive index that differs from that of the original. To generate a degree of adequate aberration to balance the aberration from a thin layer, it is necessary to estimate the amount of arising aberration correctly when a point object deviates from its aberration-free position. We analytically induce the exact form of an arising spherical aberration with an on-axis object position for general optical systems that satisfy the Abbe sine condition and express a fourth-order approximation of that form using simple parameters that are conventionally used for the aberration of a thin lens. To verify the correctness of the proposed formula, a comparison between this analysis and simulation results is applied to several sample optical systems using commercial lens-design software. PMID- 19137035 TI - Mueller matrix imaging of targets under an air-sea interface. AB - The Mueller matrix imaging method is a powerful tool for target detection. In this study, the effect of the air-sea interface on the detection of underwater objects is studied. A backward Monte Carlo code has been developed to study this effect. The main result is that the reflection of the diffuse sky light by the interface reduces the Mueller image contrast. If the air-sea interface is ruffled by wind, the distinction between different regions of the underwater target is smoothed out. The effect of the finite size of an active light source is also studied. The image contrast is found to be relatively insensitive to the size of the light source. The volume scattering function plays an important role on the underwater object detection. Generally, a smaller asymmetry parameter decreases the contrast of the polarimetry images. PMID- 19137036 TI - Coma measurement by use of an alternating phase-shifting mask mark with a specific phase width. AB - The correlation between the coma sensitivity of the alternating phase-shifting mask (Alt-PSM) mark and the mark's structure is studied based on the Hopkins theory of partially coherent imaging and positive resist optical lithography (PROLITH) simulation. It is found that an optimized Alt-PSM mark with its phase width being two-thirds its pitch has a higher sensitivity to coma than Alt-PSM marks with the same pitch and the different phase widths. The pitch of the Alt PSM mark is also optimized by PROLITH simulation, and the structure of p=1.92lambda/NA and pw=2p/3 proves to be with the highest sensitivity. The optimized Alt-PSM mark is used as a measurement mark to retrieve coma aberration from the projection optics in lithographic tools. In comparison with an ordinary Alt-PSM mark with its phase width being a half its pitch, the measurement accuracies of Z(7) and Z(14) apparently increase. PMID- 19137037 TI - Angle-dependent encoding of multiple asymmetric symbols in a binary phase hologram with a spatial segmentation. AB - A method of encoding multiple asymmetric symbols into a single thin binary Fourier hologram is presented. It assumes a combination of a spatial segmentation and carrier frequencies in order to achieve multiple reconstructed images selectable by the angle of the incident laser beam. The proper segmentation function with an optimized period allows us to encode a number of different objects with little loss of reconstruction quality. A special sequence of phase encoding steps and a binarization enable recording of asymmetric symbols into a binary hologram. The description of the design procedure is given, followed by the experimental results confirming the conclusions conceived from numerical simulations. The method can be used practically for the design of simple translucent holographic head-up displays. PMID- 19137038 TI - Exploitation of multilayer coatings for infrared surface plasmon resonance fiber sensors. AB - We demonstrate surface plasmon resonance (SPR) fiber devices based upon ultraviolet inscription of a grating-type structure into both single-layered and multilayered thin films deposited on the flat side of a lapped D-shaped fiber. The single-layered devices were fabricated from germanium, while the multilayered ones comprised layers of germanium, silica, and silver. Some of the devices operated in air with high coupling efficiency in excess of 40 dB and an estimated index sensitivity of Delta lambda/Delta n=90 nm from 1 to 1.15 index range, while others provided an index sensitivity of Delta lambda/Delta n=6790 nm for refractive indices from 1.33 to 1.37. PMID- 19137039 TI - Cascaded adaptive-mask algorithm for twin-image removal and its application to digital holograms of ice crystals. AB - An iterative Gerchberg-Saxton-type algorithm with a support constraint for twin image removal from reconstructed Gabor inline holograms of single plane objects is described. It is applied to simulated holograms and to holograms of ice crystals recorded in the laboratory and in atmospheric clouds in situ. The algorithm is characterized by a distinction between object and background region and an iterative adaption of the object mask. Applying the algorithm to recorded inline holograms of atmospheric objects, the twin-image artifacts are removed successfully, for the first time allowing for a proper access to the in situ phase information on atmospheric ice crystals. It is also demonstrated that, after application of the algorithm, previously indiscernible internal object features can become visible for large Fresnel numbers. PMID- 19137040 TI - Programmable optically reconfigurable gate array architecture and its writer. AB - Recently, optically reconfigurable gate arrays (ORGAs), which consist of a gate array VLSI, a holographic memory, and a laser array, have been developed to achieve huge virtual gate counts that vastly surpass those of currently available VLSIs. By exploiting the large storage capacity of a holographic memory, VLSIs with more than 1 teragate counts will be producible. However, compared with current field programmable gate arrays, conventional ORGAs have one important shortcoming: they cannot be reprogrammed after fabrication. To reprogram ORGAs, a holographic memory must be disassembled from its ORGA package, then reprogrammed outside of the ORGA package using a holographic memory writer. It must then be implemented onto the ORGA package with high precision techniques beyond that which can be provided by manual assembly. Therefore, to improve this shortcoming, this paper proposes what is believed to be the world's first programmable ORGA architecture with no disassembly. Finally, the availability of this architecture is discussed based on the experimental results. PMID- 19137041 TI - Effects of phase shift between two photo-alignment substrates on diffraction properties in liquid crystalline grating cells. AB - Diffractive optical devices using low-molar-mass liquid crystals are of great important for realizing numerical optical functions such as optical memory and information systems. In the present study the liquid crystalline gratings have been fabricated using a pair of photo-alignment substrates, and effects of the phase shift between the two photo-alignment substrates on the diffraction properties have been investigated. The liquid crystalline grating has been constructed with variable polarization dependence that can be controlled by the phase shift between the two photo-alignment substrates and twisted angles of the nematic directors. The theoretical considerations were also performed using the elastic continuum theory, Jones calculus, and diffraction theory. A guiding principle that controls the diffraction properties of the liquid crystalline grating cell was successfully presented. PMID- 19137042 TI - Transmittance enhancement of a wire-grid polarizer by antireflection coating. AB - We examined the effect of an antireflection (AR) coating to enhance the TM transmittance of the wire-grid polarizer. The polarization transmission spectra were calculated using the rigorous coupled-wave analysis. As a result, we verified that an AR film should be inserted between a wire-grid and a Si substrate as regards the TM transmittance and the polarization function. Based on the simulation results, we fabricated a tungsten silicide (WSi) wire-grid polarizer with SiO films on both sides of the Si substrate. The transmittance exceeded 80% at a 4-5 microm wavelength range, although the theoretical transmittance of Si substrate is 54% and the ratio of the TM and TE transmittances reached 24 dB at a 3 microm wavelength when the WSi grating has a 300 nm thickness, a 400 nm period, and a fill factor of 0.6. Wire-grid polarizers with higher transmittance and larger extinction ratio can be obtained by adjusting the AR film thickness, the fill factor, and the thickness of the WSi grating. PMID- 19137043 TI - Analysis of imaging properties of a microlens based on the method for a dyadic Green's function. AB - The dyadic Green's function (DGF) is applied to examine the effect of focal shift in a spherical microlens with the variation of the numerical aperture for a given Fresnel number when a monochromatic plane wave with x linear polarization is incident on the microlens. By comparing the results based on the method for the vector Kirchhoff diffraction theory [J. Opt. Soc. Am. A22, 68-76 (2005)], the effect of the spherical aberration on focal shift in a microlens is evaluated, and the influences of NA as well as the spherical aberration on the transverse electric energy density distribution in the focal plane are also investigated. In contrast with other vector formulations of imaging theory, which mainly focus on the focal shift in an aplanatic system, the DGF method is more practical and effective to locate the principal maximum energy density along the normal axis and to study transverse electric energy density distribution, because the actual shape of a microlens and the effects of aberrations are considered. PMID- 19137044 TI - Backscatter suppression for underwater modulating retroreflector links using polarization discrimination. AB - Free space optical links underwater have the potential to enable short range (<100 m) high-bandwidth (megabits per second) data links that have a low probability of detection and interception. The use of a retroreflecting free space optical link in water has the added advantage of allowing much of the weight and power burden of the link to remain at one end. While modulating retroreflectors have been successfully implemented in above-water links, the underwater environment introduces new challenges. The focus of this paper is to address these challenges and to investigate techniques for minimizing their effect on the link performance. PMID- 19137045 TI - Characterization of a dissolved oxygen sensor made of plastic optical fiber coated with ruthenium-incorporated solgel. AB - A dissolved oxygen sensor made of plastic optical fiber as the substrate and dichlorotris (1, 10-phenanthroline) ruthenium as a fluorescence indicator is studied. Oxygen quenching characteristics of both intensity and phase were measured; the obtained characteristics showed deviation from the linear relation described by the Stern-Volmer equation. A two-layer model is proposed to explain the deviation, and main parameters can be deduced with the model. PMID- 19137046 TI - Ultrathin four-reflection imager. AB - We present the design and experimental demonstration of an ultrathin four reflection imager. The F/1.15 prototype imager achieves a focal length of 18.6 mm in a track length of just 5.5 mm, providing a 17 degrees field of view over 1.92 megapixels of a color image sensor with 3 microm pixels. We also present the design and experimental results of pupil-phase encoding and postprocessing, which were applied to extend the depth of field and compensate a small amount of axial chromatic aberration present in the four-reflection imager prototype. PMID- 19137047 TI - Direct measurement of absorption-induced wavefront distortion in high optical power systems. AB - Wavefront distortion due to absorption in the substrates and coatings of mirrors in advanced gravitational wave interferometers has the potential to compromise the operation and sensitivity of these interferometers [Opt. Lett.29, 2635-2637 (2004)]. We report the first direct spatially-resolved measurement, to our knowledge, of such wavefront distortion in a high optical power cavity. The measurement was made using an ultrahigh sensitivity Hartmann wavefront sensor on a dedicated test facility. The sensitivity of the sensor was lambda/730, where lambda=800 nm. PMID- 19137048 TI - Linewidth reduction of a broad-area laser diode array in a compound external cavity. AB - A compound external cavity is designed and implemented to achieve a homogeneous spectrum distribution of broad area laser diodes in an array and to narrow the spectral linewidth of the entire array. The compound external cavity is composed of an optical coupler and a Littrow external cavity with a telescope along the fast axis. The inhomogeneous distribution of individual laser diodes spectrum generated by the "smile" effect was reduced by the telescope. The effective transverse coupling among individual laser diodes in an array was enhanced by the optical coupler, which further reduced the spectrum inhomogeneous distribution of the entire array. The spectral linewidth of a 49-emitter laser array is reduced to 0.1 nm at the output power of 12.5 W. PMID- 19137049 TI - High photorefractive gain at counterpropagating geometry in CdTe:Ge at 1.064 microm and 1.55 microm. AB - Recording of efficient reflection holograms is achieved in CdTe:Ge at lambda=1.064 microm and lambda=1.55 microm. The gain factor measured at both wavelengths for counterpropagating two-beam coupling considerably exceeds the absorption constant and transcends all values previously reported for semiconductors with no external field. The dependences of the gain factor on intensity and grating spacing are studied. Some crystal characteristics are estimated in the frame of a single band, one mobile species approximation of space-charge formation. The homogeneity of photorefractive properties in the crystal volume is demonstrated. PMID- 19137050 TI - Third-order optical nonlinearity of gold nanoparticle arrays embedded in a BaTiO3 matrix. AB - Au:BaTiO(3) composite films comprising hexagon-shaped Au nanoparticle arrays covered with BaTiO(3) matrix were fabricated by double-layer nanosphere lithography and pulsed laser deposition technique. The optical nonlinearity of the composite film was determined using the Z-scan technique at a wavelength of 532 nm and a pulse duration of 25 ps. The third-order nonlinear optical susceptibility, chi(3), was found to be 2.9x10(-8) esu, which is comparable with the best values in metal-dielectric films comprising randomly distributed spherical particles but with much higher metal concentrations. The local electric field enhancement in and near the particles was investigated using the dipole discrete approximation method. PMID- 19137051 TI - Experimental and theoretical inspection of the phase-to-height relation in Fourier transform profilometry. AB - The measurement of an object's shape using projected fringe patterns needs a relation between the measured phase and the object's height. Among various methods, the Fourier transform profilometry proposed by Takeda and Mutoh [Appl. Opt.22, 3977-3982 (1983)] is widely used in the literature. Rajoub et al. have shown that the reference relation given by Takeda is erroneous [J. Opt. A. Pure Appl. Opt.9, 66-75 (2007)]. This paper follows from Rajoub's study. Our results for the phase agree with Rajoub's results for both parallel- and crossed-optical axes geometries and for either collimated or noncollimated projection. Our two main results are: (i) we show experimental evidence of the error in Takeda's formula and (ii) we explain the error in Takeda's derivation and we show that Rajoub's argument concerning Takeda's error is not correct. PMID- 19137052 TI - Measurement of Gaussian laser beam radius using the knife-edge technique: improvement on data analysis. AB - We revisited the well known Khosrofian and Garetz inversion algorithm [Appl. Opt.22, 3406-3410 (1983)] that was developed to analyze data obtained by the application of the traveling knife-edge technique. We have analyzed the approximated fitting function that was used for adjusting their experimental data and have found that it is not optimized to work with a full range of the experimentally-measured data. We have numerically calculated a new set of coefficients, which makes the approximated function suitable for a full experimental range, considerably improving the accuracy of the measurement of a radius of a focused Gaussian laser beam. PMID- 19137053 TI - Dual-prism interferometer for collimation testing. AB - An air-wedge lateral-shear interferometer using two prisms is presented. With a variable shear, the interferometer is suitable for testing collimation of a wide range of beam sizes down to a few millimeters in diameter. No antireflection coatings are necessary. Collimation for a light source with short coherent length is also demonstrated. PMID- 19137054 TI - Harmonic wavelet analysis of modulated tunable diode laser absorption spectroscopy signals. AB - Wavelet analyses of tunable diode laser absorption spectroscopy signals were performed. The absorption spectroscopy data were obtained by repeatedly scanning the beams from a tunable diode laser operating in the near infrared across absorption lines of gaseous NH(3) contained within a windowed glass tube. The laser was modulated and wavelet analyses of the absorption data were performed. It was observed that harmonic wavelets could simultaneously extract the 1f and 2f harmonics as well as higher-order harmonics from the direct absorption data. PMID- 19137055 TI - Absorption and wavelength modulation spectroscopy of NO2 using a tunable, external cavity continuous wave quantum cascade laser. AB - The absorption spectra and wavelength modulation spectroscopy (WMS) of NO(2) using a tunable, external cavity CW quantum cascade laser operating at room temperature in the region of 1625 to 1645 cm(-1) are reported. The external cavity quantum cascade laser enabled us to record continuous absorption spectra of low concentrations of NO(2) over a broad range (approximately 16 cm(-1)), demonstrating the potential for simultaneously recording the complex spectra of multiple species. This capability allows the identification of a particular species of interest with high sensitivity and selectivity. The measured spectra are in excellent agreement with the spectra from the high-resolution transmission molecular absorption database [J. Quant. Spectrosc. Radiat. Transfer 96, 139-204 (2005)]. We also conduct WMS for the first time using an external cavity quantum cascade laser, a technique that enhances the sensitivity of detection. By employing WMS, we could detect low-intensity absorption lines, which are not visible in the simple absorption spectra, and demonstrate a minimum detection limit at the 100 ppb level with a short-path absorption cell. Details of the tunable, external cavity quantum cascade laser system and its performance are discussed. PMID- 19137056 TI - Optical encoder based on a nondiffractive beam II. AB - In a previous work, we introduced the design of an optical encoder based on a nondiffractive beam and demonstrated that it generates a suitable output sinusoidal signal [Appl. Opt.47, 2201-2206 (2008)]. In this work, we experimentally, analytically, and numerically study the dependence of the system performance on its parameters (grating pitch, photodetector size, etc.) and propose three different optimization criteria for which the tolerance to variations in the system parameters is also analyzed. We conclude that the proposed design generates a suitable output signal, with high contrast and very low harmonic distortion, while having a remarkable tolerance to variations in its parameters and to mechanical perturbations. PMID- 19137057 TI - Tilted parallel dielectric slab as a multilevel attenuator for incident p- or s polarized light. AB - Under the condition of first-order blooming, a parallel dielectric slab, which is inserted in the path of an obliquely incident p- or s-polarized light beam, introduces multiple discrete attenuation levels given by 1/3, 4/27, 4/243,...... in reflection and 4/9, 4/81, 4/729,...... in transmission. These attenuation levels are independent of the slab refractive index, incident p or s linear polarization, or the presence of identical transparent surface coatings at the front and back sides of the slab. Therefore, the tilted slab provides multidecade reflectance and attenuation reference values that can be used in calibrating spectrophotometers and filters, and also for testing the linearity of photodetectors. For an uncoated dielectric slab, incidence angles that cause first-order blooming are determined as functions of the slab refractive index for incident p- or s-polarized light. PMID- 19137058 TI - Formulation of frequency stability limited by laser intrinsic noise in feedback systems. AB - We investigated the influence of amplitude modulation (AM) noise and phase modulation (PM) noise of a laser source on the frequency stability in frequency stabilization systems. We estimated the frequency stability and evaluated the efficacy of a noise reduction technique (the Doppler-trend subtraction method) of a laser diode frequency stabilization system, where enhanced intensity noise arising from PM-to-AM noise conversion through a reference gas cell is reduced using the technique employed in modulation transfer spectroscopy. To evaluate the relationship between the laser's intrinsic noise and its frequency stability, we performed noise spectrum measurements and formulated frequency stability in addition to measuring Allan standard deviation. As a result, it is found that the extra noise generated in PM-to-AM conversion is efficiently removed by the Doppler-trend subtraction method and that within the feedback bandwidth, the frequency stability becomes 1 order of magnitude better than that without the method. PMID- 19137059 TI - Simultaneous Confidence Intervals Based on the Percentile Bootstrap Approach. AB - This note concerns the construction of bootstrap simultaneous confidence intervals (SCI) for m parameters. Given B bootstrap samples, we suggest an algorithm with complexity of O(mB log(B)). We apply our algorithm to construct a confidence region for time dependent probabilities of progression in multiple sclerosis and for coefficients in a logistic regression analysis. Alternative normal based simultaneous confidence intervals are presented and compared to the bootstrap intervals. PMID- 19137060 TI - Metal Enhanced Fluorescence on Silicon Wafer Substrates. AB - We report on the fluorescence enhancement induced by silver island film (SIF) deposited on a silicon wafer. The model immunoassay was studied on silvered and unsilvered wafers. The fluorescence brightness of Rhodamine Red X increased about 300% on the SIF, while the lifetime was reduced by several fold and the photostability increased substantially. We discuss potential uses of silicon wafer substrates in multiplex assays in which the fluorescence is enhanced due to the SIF, and the multiplexing is achieved by using micro transponders. PMID- 19137061 TI - Cationic nano-copolymers mediated IKKbeta targeting siRNA inhibit the proliferation of human Tenon's capsule fibroblasts in vitro. AB - PURPOSE: To synthesize a ternary cationic copolymer called CS-g-(PEI-b-mPEG) and characterize its features as a non-viral siRNA carrier; in turn, to investigate the influence of small interfering RNA (siRNA) targeting IkappaB kinase subunit beta (IKKbeta) on the proliferation of human Tenon's capsule fibroblasts (HTFs) in vitro. METHODS: First, a novel cationic copolymer composed of low molecular weight, linear poly(ethyleneimine) [PEI] blocked with polyethylene glycol (PEG) and grafted onto a chitosan (CS) molecule was synthesized. CS-g-(PEI-b-mPEG) was then compacted with 21nt siRNA at various copolymer/siRNA charge (N/P) ratios, and the resulting complexes were characterized by dynamic light scattering, gel electrophoresis, and serum incubation. Cell Titer 96 AQ(ueous) One Solution cell proliferation assay was used to investigate the cytotoxicity of this cationic copolymer. Second, siRNAs targeting IKKbeta (IKKBeta-siRNAs) were delivered into the HTFs using CS-g-(PEI-b-mPEG) as the vehicle. Real-time reverse transcription polymerase chain reaction (RT-PCR) subsequently assessed the mRNA level of IKKbeta, and western blot assay was used to determine protein expression. After IKKB-siRNA transfection, Cell Titer 96 AQ(ueous) One Solution cell proliferation assay was used to evaluate the proliferation of HTFs. RESULTS: The diameter of the CS-g-(PEI-b-mPEG)/siRNA complexes tended to decrease whereas their zeta potential tended to increase as the N/P ratio increased. The CS-g-(PEI-b-mPEG) copolymer showed good siRNA binding ability and high siRNA protection capacity. Furthermore, the copolymer presented remarkable transfection efficiency and showed much less cytotoxicity than 25 kDa PEI. IKKB-siRNAs were successfully delivered into HTFs using CS-g-(PEI-b-mPEG) as a vector. As a result, the expression of IKKbeta was downregulated at both the mRNA and protein levels, and the activation of nuclear factor-kappaB (NF-kappaB) in the HTFs was subsequently inhibited. Most impressively, the proliferation of HTFs was also effectively suppressed through the blocking of the NF-kappaB pathway. CONCLUSIONS: All the results demonstrate that CS-g-(PEI-b-mPEG) is a promising candidate for siRNA delivery, featuring excellent biocompatibility, biodegradability, and transfection efficiency. The RNA interference (RNAi) strategy using cationic copolymers as siRNA carriers will be a safe and efficient anti-scarring method following glaucoma filtration surgery. PMID- 19137062 TI - Accumulation of phosphorylated beta-catenin enhances ROS-induced cell death in presenilin-deficient cells. AB - Presenilin (PS) is involved in many cellular events under physiological and pathological conditions. Previous reports have revealed that PS deficiency results in hyperproliferation and resistance to apoptotic cell death. In the present study, we investigated the effects of PS on beta-catenin and cell mortality during serum deprivation. Under these conditions, PS1/PS2 double knockout MEFs showed aberrant accumulation of phospho-beta-catenin, higher ROS generation, and notable cell death. Inhibition of beta-catenin phosphorylation by LiCl reversed ROS generation and cell death in PS deficient cells. In addition, the K19/49R mutant form of beta-catenin, which undergoes normal phosphorylation but not ubiquitination, induced cytotoxicity, while the phosphorylation deficient S37A beta-catenin mutant failed to induce cytotoxicity. These results indicate that aberrant accumulation of phospho-beta-catenin underlies ROS-mediated cell death in the absence of PS. We propose that the regulation of beta-catenin is useful for identifying therapeutic targets of hyperproliferative diseases and other degenerative conditions. PMID- 19137063 TI - Immunohistochemical characterisation of cell-type specific expression of CK1delta in various tissues of young adult BALB/c mice. AB - BACKGROUND: Casein kinase 1 delta (CK1delta) phosphorylates many key proteins playing important roles in such biological processes as cell growth, differentiation, apoptosis, circadian rhythm and vesicle transport. Furthermore, deregulation of CK1delta has been linked to neurodegenerative diseases and cancer. In this study, the cell specific distribution of CK1delta in various tissues and organs of young adult BALB/c mice was analysed by immunohistochemistry. METHODOLOGY/PRINCIPAL FINDINGS: Immunohistochemical staining of CK1delta was performed using three different antibodies against CK1delta. A high expression of CK1delta was found in a variety of tissues and organ systems and in several cell types of endodermal, mesodermal and ectodermal origin. CONCLUSIONS: These results give an overview of the cell-type specific expression of CK1delta in different organs under normal conditions. Thus, they provide evidence for possible cell-type specific functions of CK1delta, where CK1delta can interact with and modulate the activity of key regulator proteins by site directed phosphorylation. Furthermore, they provide the basis for future analyses of CK1delta in these tissues. PMID- 19137064 TI - Interleukin-32 promotes osteoclast differentiation but not osteoclast activation. AB - BACKGROUND: Interleukin-32 (IL-32) is a newly described cytokine produced after stimulation by IL-2 or IL-18 and IFN-gamma. IL-32 has the typical properties of a pro-inflammatory mediator and although its role in rheumatoid arthritis has been recently reported its effect on the osteoclastogenesis process remains unclear. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we have shown that IL-32 was a potent modulator of osteoclastogenesis in vitro, whereby it promoted the differentiation of osteoclast precursors into TRAcP+ VNR+ multinucleated cells expressing specific osteoclast markers (up-regulation of NFATc1, OSCAR, Cathepsin K), but it was incapable of inducing the maturation of these multinucleated cells into bone-resorbing cells. The lack of bone resorption in IL-32-treated cultures could in part be explain by the lack of F-actin ring formation by the multinucleated cells generated. Moreover, when IL-32 was added to PBMC cultures maintained with soluble RANKL, although the number of newly generated osteoclast was increased, a significant decrease of the percentage of lacunar resorption was evident suggesting a possible inhibitory effect of this cytokine on osteoclast activation. To determine the mechanism by which IL-32 induces such response, we sought to determine the intracellular pathways activated and the release of soluble mediators in response to IL-32. Our results indicated that compared to RANKL, IL-32 induced a massive activation of ERK1/2 and Akt. Moreover, IL-32 was also capable of stimulating the release of IL-4 and IFN-gamma, two known inhibitors of osteoclast formation and activation. CONCLUSIONS/SIGNIFICANCE: This is the first in vitro report on the complex role of IL-32 on osteoclast precursors. Further clarification on the exact role of IL-32 in vivo is required prior to the development of any potential therapeutic approach. PMID- 19137065 TI - Structural model of the Rev regulatory protein from equine infectious anemia virus. AB - Rev is an essential regulatory protein in the equine infectious anemia virus (EIAV) and other lentiviruses, including HIV-1. It binds incompletely spliced viral mRNAs and shuttles them from the nucleus to the cytoplasm, a critical prerequisite for the production of viral structural proteins and genomic RNA. Despite its important role in production of infectious virus, the development of antiviral therapies directed against Rev has been hampered by the lack of an experimentally-determined structure of the full length protein. We have used a combined computational and biochemical approach to generate and evaluate a structural model of the Rev protein. The modeled EIAV Rev (ERev) structure includes a total of 6 helices, four of which form an anti-parallel four-helix bundle. The first helix contains the leucine-rich nuclear export signal (NES). An arginine-rich RNA binding motif, RRDRW, is located in a solvent-exposed loop region. An ERLE motif required for Rev activity is predicted to be buried in the core of modeled structure where it plays an essential role in stabilization of the Rev fold. This structural model is supported by existing genetic and functional data as well as by targeted mutagenesis of residues predicted to be essential for overall structural integrity. Our predicted structure should increase understanding of structure-function relationships in Rev and may provide a basis for the design of new therapies for lentiviral diseases. PMID- 19137066 TI - Studying early lethality of 45,XO (Turner's syndrome) embryos using human embryonic stem cells. AB - Turner's syndrome (caused by monosomy of chromosome X) is one of the most common chromosomal abnormalities in females. Although 3% of all pregnancies start with XO embryos, 99% of these pregnancies terminate spontaneously during the first trimester. The common genetic explanation for the early lethality of monosomy X embryos, as well as the phenotype of surviving individuals is haploinsufficiency of pseudoautosomal genes on the X chromosome. Another possible mechanism is null expression of imprinted genes on the X chromosome due to the loss of the expressed allele. In contrast to humans, XO mice are viable, and fertile. Thus, neither cells from patients nor mouse models can be used in order to study the cause of early lethality in XO embryos. Human embryonic stem cells (HESCs) can differentiate in culture into cells from the three embryonic germ layers as well as into extraembryonic cells. These cells have been shown to have great value in modeling human developmental genetic disorders. In order to study the reasons for the early lethality of 45,XO embryos we have isolated HESCs that have spontaneously lost one of their sex chromosomes. To examine the possibility that imprinted genes on the X chromosome play a role in the phenotype of XO embryos, we have identified genes that were no longer expressed in the mutant cells. None of these genes showed a monoallelic expression in XX cells, implying that imprinting is not playing a major role in the phenotype of XO embryos. To suggest an explanation for the embryonic lethality caused by monosomy X, we have differentiated the XO HESCs in vitro an in vivo. DNA microarray analysis of the differentiated cells enabled us to compare the expression of tissue specific genes in XO and XX cells. The tissue that showed the most significant differences between the clones was the placenta. Many placental genes are expressed at much higher levels in XX cells in compare to XO cells. Thus, we suggest that abnormal placental differentiation as a result of haploinsufficiency of X-linked pseudoautosomal genes causes the early lethality in XO human embryos. PMID- 19137067 TI - Augmented lung inflammation protects against influenza A pneumonia. AB - BACKGROUND: Influenza pneumonia causes high mortality every year, and pandemic episodes kill millions of people. Influenza-related mortality has been variously ascribed to an ineffective host response that fails to limit viral replication, an excessive host inflammatory response that results in lung injury and impairment of gas exchange, or to bacterial superinfection. We sought to determine whether lung inflammation promoted or impaired host survival in influenza pneumonia. METHODS AND FINDINGS: To distinguish among these possible causes of influenza-related death, we induced robust lung inflammation by exposing mice to an aerosolized bacterial lysate prior to challenge with live virus. The treatment induced expression of the inflammatory cytokines IL-6 and TNF in bronchoalveolar lavage fluid 8- and 40-fold greater, respectively, than that caused by lethal influenza infection. Yet, this augmented inflammation was associated with striking resistance to host mortality (0% vs 90% survival, p = 0.0001) and reduced viral titers (p = 0.004). Bacterial superinfection of virus infected lungs was not observed. When mice were repeatedly exposed to the bacterial lysate, as would be clinically desirable during an influenza epidemic, there was no tachyphylaxis of the induced viral resistance. When the bacterial lysate was administered after the viral challenge, there was still some mortality benefit, and when ribavirin was added to the aerosolized bacterial lysate, host survival was synergistically improved (0% vs 93.3% survival, p<0.0001). CONCLUSIONS: Together, these data indicate that innate immune resistance to influenza can be effectively stimulated, and suggest that ineffective rather than excessive inflammation is the major cause of mortality in influenza pneumonia. PMID- 19137068 TI - Granular assembly of alpha-synuclein leading to the accelerated amyloid fibril formation with shear stress. AB - alpha-Synuclein participates in the Lewy body formation of Parkinson's disease. Elucidation of the underlying molecular mechanism of the amyloid fibril formation is crucial not only to develop a controlling strategy toward the disease, but also to apply the protein fibrils for future biotechnology. Discernable homogeneous granules of alpha-synuclein composed of approximately 11 monomers in average were isolated in the middle of a lag phase during the in vitro fibrillation process. They were demonstrated to experience almost instantaneous fibrillation during a single 12-min centrifugal membrane-filtration at 14,000 x g. The granular assembly leading to the drastically accelerated fibril formation was demonstrated to be a result of the physical influence of shear force imposed on the preformed granular structures by either centrifugal filtration or rheometer. Structural rearrangement of the preformed oligomomeric structures is attributable for the suprastructure formation in which the granules act as a growing unit for the fibril formation. To parallel the prevailing notion of nucleation-dependent amyloidosis, we propose a double-concerted fibrillation model as one of the mechanisms to explain the in vitro fibrillation of alpha synuclein, in which two consecutive concerted associations of monomers and subsequent oligomeric granular species are responsible for the eventual amyloid fibril formation. PMID- 19137069 TI - A novel peptide enhances therapeutic efficacy of liposomal anti-cancer drugs in mice models of human lung cancer. AB - Lung cancer is the leading cause of cancer-related mortality worldwide. The lack of tumor specificity remains a major drawback for effective chemotherapies and results in dose-limiting toxicities. However, a ligand-mediated drug delivery system should be able to render chemotherapy more specific to tumor cells and less toxic to normal tissues. In this study, we isolated a novel peptide ligand from a phage-displayed peptide library that bound to non-small cell lung cancer (NSCLC) cell lines. The targeting phage bound to several NSCLC cell lines but not to normal cells. Both the targeting phage and the synthetic peptide recognized the surgical specimens of NSCLC with a positive rate of 75% (27 of 36 specimens). In severe combined immunodeficiency (SCID) mice bearing NSCLC xenografts, the targeting phage specifically bound to tumor masses. The tumor homing ability of the targeting phage was inhibited by the cognate synthetic peptide, but not by a control or a WTY-mutated peptide. When the targeting peptide was coupled to liposomes carrying doxorubicin or vinorelbine, the therapeutic index of the chemotherapeutic agents and the survival rates of mice with human lung cancer xenografts markedly increased. Furthermore, the targeting liposomes increased drug accumulation in tumor tissues by 5.7-fold compared with free drugs and enhanced cancer cell apoptosis resulting from a higher concentration of bioavailable doxorubicin. The current study suggests that this tumor-specific peptide may be used to create chemotherapies specifically targeting tumor cells in the treatment of NSCLC and to design targeted gene transfer vectors or it may be used one in the diagnosis of this malignancy. PMID- 19137070 TI - Retraction and remodeling of rod spherules are early events following experimental retinal detachment: an ultrastructural study using serial sections. AB - PURPOSE: To describe changes induced by retinal detachment in the ultrastructure and organization of rod terminals and their connections with B-type horizontal cell (HC) axon terminals and rod bipolar cell (RB) dendrites. METHODS: Sections from control, 3 day, 7 day, and 28 day detached feline retinas were prepared for confocal immunofluorescence, light microscopy, and electron microscopy (EM). In addition, 100 mum-thick vibratome sections were immunolabeled with markers for photoreceptor terminals, HCs, and RBs. More than 40 rod spherules were studied in 90 nm-thick serial sections by transmission EM to greater detail changes in their ultrastructure and innervation. RESULTS: Following retinal detachment, many rod terminals retracted varying distances toward their respective cell bodies in the outer nuclear layer (ONL). In retinas detached for 1 to 4 weeks, an altered synaptic vesicle population and associated ribbons were found in all retracting terminals. Many rod somata in the distal ONL seemed to lack synaptic terminal structures altogether. In a retina detached for 1 week, EM showed that less than half of the retracted terminals remain in contact with RB dendrites. In contrast, almost every surviving spherule was contacted by neurite outgrowths from the axon terminals of the B-type HC. Although retracted spherules had several presynaptic structures similar to those in normal retina, numerous changes occurred in their overall synaptic architecture. The spherule's invagination was shallower, contained fewer postsynaptic processes, and often had "opened," allowing swollen HC processes apposing the synaptic ribbon to directly contact other processes of the outer plexiform layer (OPL) neuropil. Whereas in normal cat retina each HC "lobe" comes from a different axon terminal system, after detachment, the opposing lateral elements can stem from the same terminal. The innervating RB dendrites that branched off stout RB dendritic trunks that extended up into the ONL were thinner than normal, unbranched, often electron dense, and lacked organelles. When present, most merely lay adjacent to retracting spherules rather than enter any synaptic invagination that might still occur. CONCLUSIONS: Immunocytochemistry enabled RB and HC neurites to appear postsynaptic to retracted rod terminals. However, at the ultrastructural level, HCs seemed to more consistently retain connection with the retracted spherules than the RBs. The highly conserved architecture of the rod spherule was lost as the invagination opened and postsynaptic contacts became fewer. It would seem that the lack of RB central elements as well as the drastic alterations in the architecture of most retracted terminals would necessarily alter the physiology of this complex synapse. PMID- 19137071 TI - Preservation of intact adult rat photoreceptors in vitro: study of dissociation techniques and the effect of light. AB - PURPOSE: Intact adult photoreceptors in culture can be a valuable tool in the search of therapies for retinal degenerations. The major challenge in this technique is that photoreceptors undergo an alteration in cytoarchitecture and loss of outer segment during the cell culture process. This study compared techniques for the isolation of photoreceptor cells from adult rat retinas to determine which technique yields the highest percent of structurally well preserved cells in vitro. In addition, the role of light exposure during the dissociation and culture process was investigated to minimize photoreceptor cell deformation over time in culture. METHODS: Photoreceptor cells from adult rat retinas were isolated and quantified using three dissociation techniques: enzymatic dissociation with gentle pipeting; enzymatic dissociation with gentle pipeting and centrifugation; and non-enzymatic dissociation with gentle pipeting. To evaluate the effect of light exposure on cell deformation, we performed dissociations and cell seeding both in dark- and light-adapted conditions and measured the deformation of photoreceptors over a 12 h period right after dissociation. Cell viability in both conditions was evaluated after 4 and 7 days in culture. Preservation of cell structure in culture was assessed by immunofluorescence labeling of cells with anti-rhodopsin and 4',6-diamidino-2 phenylindole (DAPI) nuclear staining. RESULTS: An enzymatic technique followed by gentle pipeting or mechanical trituration yielded the highest number of intact elongated photoreceptors right after dissociation. Data suggested that centrifugation after the dissociation contributed to cell deformation immediately after isolation. Immunohistochemistry results showed that cells had deformed into a circular shape by 2 days after seeding. However, photoreceptors isolated in dark conditions maintained their elongated shape, even 7 days after seeding. Performing experiments in dark also promoted a higher number of cells to remain viable with time. CONCLUSIONS: The current study demonstrated the importance of proper isolation techniques to obtain the maximum amount of intact photoreceptor cells. The data suggested that a gentle dissociation technique, consisting of enzymatic treatment followed by moderate pipeting of the retinal tissue, may be the key to obtain a high number of intact or structurally preserved photoreceptors. Furthermore, isolation and cell culture procedures performed under dark conditions may facilitate to maintain high number of elongated photoreceptor cells in vitro. PMID- 19137072 TI - Divergent effects of PERK and IRE1 signaling on cell viability. AB - Protein misfolding in the endoplasmic reticulum (ER) activates a set of intracellular signaling pathways, collectively termed the Unfolded Protein Response (UPR). UPR signaling promotes cell survival by reducing misfolded protein levels. If homeostasis cannot be restored, UPR signaling promotes cell death. The molecular basis for the switch between prosurvival and proapoptotic UPR function is poorly understood. The ER-resident proteins, PERK and IRE1, control two key UPR signaling pathways. Protein misfolding concomitantly activates PERK and IRE1 and has clouded insight into their contributions toward life or death cell fates. Here, we employed chemical-genetic strategies to activate individually PERK or IRE1 uncoupled from protein misfolding. We found that sustained PERK signaling impaired cell proliferation and promoted apoptosis. By contrast, equivalent durations of IRE1 signaling enhanced cell proliferation without promoting cell death. These results demonstrate that extended PERK and IRE1 signaling have opposite effects on cell viability. Differential activation of PERK and IRE1 may determine life or death decisions after ER protein misfolding. PMID- 19137073 TI - Promoter methylation in cytology specimens as an early detection marker for esophageal squamous dysplasia and early esophageal squamous cell carcinoma. AB - The incidence of esophageal squamous cell carcinoma (ESCC) is very high in northern China. This cancer has a very poor prognosis, mostly because it is usually diagnosed at a late stage. Detection at an earlier stage can dramatically improve prognosis. Microscopic evaluation of esophageal balloon cytology (EBC) specimens has been the most common method for early detection of ESCC, but this technique is limited by low sensitivity and specificity. The use of molecular markers may improve these screening characteristics. This study evaluates whether measurement of gene methylation in EBC specimens may have utility for the detection of esophageal squamous dysplasia and early ESCC. We evaluated the presence of methylation in eight genes shown to be methylated in ESCC in previous studies in EBC specimens from 147 patients with endoscopic biopsy diagnoses ranging from normal mucosa to severe squamous dysplasia. Methylation status was determined using quantitative methylation-specific PCR techniques. The sensitivity and specificity of methylation of each individual gene and of combinations of these genes to detect biopsyproven high-grade (moderate or severe) squamous dysplasia were determined. For individual genes, the sensitivities ranged from 9% to 34% and the specificities ranged from 77% to 99%. Using a panel of four genes (AHRR, p16INK4a, MT1G, and CLDN3) resulted in sensitivity and specificity of 50% and 68%, respectively. This study suggests that evaluation of gene methylation in EBC samples may have utility for early detection of esophageal squamous dysplasia and early ESCC; however, identification of more sensitive methylation markers will be required for development of a clinically useful screening test. PMID- 19137074 TI - Contextual analysis of fluid intelligence. AB - The nature of fluid intelligence was investigated by identifying variables that were, and were not, significantly related to this construct. Relevant information was obtained from three sources: re-analyses of data from previous studies, a study in which 791 adults performed storage-plus-processing working memory tasks, and a study in which 236 adults performed a variety of working memory, updating, and cognitive control tasks. The results suggest that fluid intelligence represents a broad individual difference dimension contributing to diverse types of controlled or effortful processing. The analyses also revealed that very few of the age-related effects on the target variables were statistically independent of effects on established cognitive abilities, which suggests most of the age related influences on a wide variety of cognitive control variables overlap with age-related influences on cognitive abilities such as fluid intelligence, episodic memory, and perceptual speed. PMID- 19137076 TI - Tracing the Early Development of Harmful Algal Blooms on the West Florida Shelf with the Aid of Lagrangian Coherent Structures. AB - Several theories have been proposed to explain the development of harmful algal blooms (HABs) produced by the toxic dinoflagellate Karenia brevis on the West Florida Shelf. However, because the early stages of HAB development are usually not detected, these theories have been so far very difficult to verify. In this paper we employ simulated Lagrangian coherent structures (LCSs) to trace potential early locations of the development of a HAB in late 2004 before it was transported to a region where it could be detected by satellite imagery. The LCSs, which are extracted from surface ocean currents produced by a data assimilative HYCOM (HYbrid-Coordinate Ocean Model) simulation, constitute material fluid barriers that demarcate potential pathways for HAB evolution. Using a simplified population dynamics model we infer the factors that could possibly lead to the development of the HAB in question. The population dynamics model determines nitrogen in two components, nutrients and phytoplankton, which are assumed to be passively advected by surface ocean currents produced by the above HYCOM simulation. Two nutrient sources are inferred for the HAB whose evolution is found to be strongly tied to the simulated LCSs. These nutrient sources are found to be located nearshore and possibly due to land runoff. PMID- 19137075 TI - Norrin attenuates protease-mediated death of transformed retinal ganglion cells. AB - PURPOSE: To investigate the effects of norrin, a nonconventional ligand for Wingless-Int (Wnt)-beta-catenin signaling pathway, on protease-mediated death of transformed rat retinal ganglion cells (RGC-5). METHODS: Transformed RGC-5 cells were treated with 2.0 microM staurosporine (SS), a broad-spectrum protein kinase C inhibitor, to induce growth arrest, differentiation, and elevated levels of tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA). RGC 5 cells were also treated with 2.0 microM SS and varying doses of recombinant norrin (3.125 to 100 ng/ml). Activation of Wnt pathway was assessed by nuclear translocation of beta-catenin. Proteolytic activity of tPA and uPA was determined by zymography assays and cell viability was determined by 3-[4,5-dimethylthiazol 2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays. Expression and phosphorylation of the low-density lipoprotein-related receptor-1 (LRP-1), a cell surface receptor for tPA and uPA, was determined by immunoprecipitation and western blot analysis. RESULTS: Compared to RGC-5 cells left untreated, cells treated with either SS alone or SS and norrin secreted elevated levels of tPA and uPA. A significant number of RGC-5 cells treated with only SS underwent cell death, whereas cells treated with SS and norrin did not, even though RGC-5 cells secreted elevated levels of tPA and uPA under both treatment conditions. Although norrin activated the Wnt pathway, Dickkopf related protein 1 (Dkk1), an inhibitor of Wnt/beta-catenin pathway, failed to completely block norrin's neuroprotective effects. Assays for expression and phosphorylation of LRP-1 indicated that tPA and uPA cause RGC-5 cell death, in part, by reducing phosphorylation of LRP-1, whereas norrin attenuated tPA and uPA-mediated RGC cell death, in part, by restoring phosphorylation of LRP-1. CONCLUSIONS: Our results suggest that norrin attenuates tPA- and uPA-mediated death of RGC-5 cells by activating Wnt/beta catenin pathway and by regulating phosphorylation of LRP-1. PMID- 19137077 TI - Identification of a novel splice-site mutation in MIP in a Chinese congenital cataract family. AB - PURPOSE: To map the locus and identify the gene causing autosomal dominant congenital cataract (ADCC) with "snail-like" phenotype in a large Chinese family. METHODS: Clinical and ophthalmologic examinations were conducted on family members and documented by slit lamp photography. Linkage analysis was performed with an initial 41 microsatellite markers, then 3 additional markers flanking the major intrinsic protein (MIP) gene. Mutations were screened by DNA sequencing and verified by restriction fragment length polymorphism (RFLP) analysis. RESULTS: Significant two-point LOD scores were obtained at 5 markers flanking MIP with the highest 3.08 (theta=0.00) at marker D12S1632. Mutation screening of MIP identified a heterozygous G>A transition at the acceptor splice site of intron 3 (IVS3 -1 G>A), abolishing a BstSF I restriction site in one allele of all the affected individuals. CONCLUSIONS: We identified a novel splice-site mutation (IVS3 -1 G>A in MIP) in a Chinese ADCC family. To our knowledge, this is the first report on an acceptor splice-site mutation in human genes associated with ADCC. PMID- 19137078 TI - Emergent Clipping without Prophylactic Decompressive Craniectomy in Patients with a Large Aneurysmal Intracerebral Hematoma. AB - OBJECTIVE: Many vascular neurosurgeons tend to remove bone flap in patients with large aneurysmal intracerebral hematomas (ICH). However, relatively little work has been done regarding the effectiveness of prophylactic decompressive craniectomy in a patient with a large aneurysmal ICH. METHODS: Large ICH was defined as hematoma when its volume exceeded 25 mL, ipsilateral to aneurysms. The patients were divided into two groups; aneurysmal subarachnoid hemorrhage (SAH) associated with large ICH, January, 1994 - December, 1999 (Group A, 41 patients), aneurysmal SAH associated with large ICH, January, 2000 - May, 2005 (Group B, 27 patients). Demographic and clinical variables including age, sex, hypertension, vasospasm, rebleeding, Hunt-Hess grade, aneurysm location, aneurysm size, and outcome were compared between two groups, and also compared between craniotomy and craniectomy patients in Group A. RESULTS: In Group A, 21 of 41 patients underwent prophylactic decompressive craniectomy. In Group B, only two patients underwent craniectomy. Surgical outcome in Group A (good 23, poor 18) was statistically not different from Group B (good 15, poor 12). Surgical outcomes between craniectomy (good 12, poor 9) and craniotomy cases (good 11, poor 9) in Group A were also comparable. CONCLUSION: We recommend that a craniotomy can be carried out safely without prophylactic craniectomy in patients with a large aneurysmal ICH if intracranial pressure is controllable with hematoma evacuation. PMID- 19137079 TI - Gamma knife radiosurgery for ten or more brain metastases. AB - OBJECTIVE: This study was performed to assess the efficacy of GKS in patients with ten or more brain metastases. METHODS: From Aug 2002 to Dec 2007, twenty-six patients (13 men and 13 women) with ten or more cerebral metastatic lesions underwent GKS. The mean age was 55 years (32-80). All patients had Karnofsky performance status (KPS) score of 70 or better. According to recursive partitioning analysis (RPA) classification, 3 patients belonged to class I and 23 to class II. The location of primary tumor was lung (21), breast (3) and unknown (2). The mean number of the lesions per patient was 16.6 (10-37). The mean cumulated volume was 10.9 cc (1.0-42.2). The median marginal dose was 15 Gy (9 23). Overall survival and the prognostic factors for the survival were retrospectively analyzed by using Kaplan Meier method and univariate analysis. RESULTS: Overall median survival from GKS was 34 weeks (8-199). Local control was possible for 79.5% of the lesions and control of all the lesions was possible in at least 14 patients (53.8%) until 6 months after GKS. New lesions appeared in 7 (26.9%) patients during the same period. At the last follow-up, 18 patients died; 6 (33.3%) from systemic causes, 10 (55.6%) from neurological causes, and 2 (11.1%) from unknown causes. Synchronous onset in non-small cell lung cancer (p=0.007), high KPS score (>/=80, p=0.029), and controlled primary disease (p=0.020) were favorable prognostic factors in univariate analysis. CONCLUSION: In carefully selected patients, GKS may be a treatment option for ten or more brain metastases. PMID- 19137080 TI - Spinal dural arteriovenous fistulas: clinical experience with endovascular treatment as a primary therapeutic modality. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy of endovascular therapy as a primary treatment for spinal dural arteriovenous fistula (DAVF). METHODS: The authors reviewed 18 patients with spinal DAVFs for whom endovascular therapy was considered as an initial treatment at a single institute between 1993 and 2006. NBCA embolization was considered the primary treatment of choice, with surgery reserved for patients in whom endovascular treatment failed. RESULTS: Surgery was performed as the primary treatment in one patient because the anterior spinal artery originated from the same arterial pedicle as the artery feeding the fistula. Embolization was used as the primary treatment modality in 17 patients, with an initial success rate of 82.4%. Two patients with incomplete embolization had to undergo surgery. One patient underwent multiple embolizations, which failed to completely occlude the fistula but relieved the patient's symptoms. Spinal DAVF recurred in two patients (one collateral development and one recanalization) during the follow-up period. The collateral development was obliterated by repeated embolization, but the patient with recanalization refused further treatment. The overall clinical status improved in 15 patients (83.3%) during the follow-up period. CONCLUSION: Endovascular therapy can be successfully used as a primary treatment for the majority of patients with spinal DAVFs. Although it is difficult to perform in some patients, endovascular embolization should be the primary treatment of choice for spinal DAVF. PMID- 19137081 TI - Radiologic Assessment of Subsidence in Stand-Alone Cervical Polyetheretherketone (PEEK) Cage. AB - OBJECTIVE: Aim of study was to find a proper method for assessing subsidence using a radiologic measurement following anterior cervical discectomy and fusion (ACDF) with stand-alone polyetheretherketone (PEEK), Solistrade mark cage. METHODS: Forty-two patients who underwent ACDF with Solistrade mark cage were selected. With a minimum follow-up of 6 months, the retrospective investigation was conducted for 37 levels in 32 patients. Mean follow-up period was 18.9 months. Total intervertebral height (TIH) of two fused vertebral bodies was measured on digital radiographs with built-in software. Degree of subsidence (DeltaTIH) was reflected by the difference between the immediate postoperative and follow-up TIH. Change of postoperative disc space height (CT-MRDeltaTIH) was reflected by the difference between TIH of the preoperative mid-sagittal 2D CT and that of the preoperative mid-sagittal T1-weighted MRI. RESULTS: Compared to preoperative findings, postoperative disc height was increased in all cases and subsidence was observed only in 3 cases. For comparison of subsidence and non subsidence group, TIH and CT-MRDeltaTIH of each group were analyzed. There was no statistically significant difference in TIH and CT-MRDeltaTIH between each group at 4 and 8 weeks, but a difference was observed at the last follow-up TIH (p=0.0497). CONCLUSION: ACDF with Solistrade mark cage was associated with relatively good radiologic long-term results. Fusion was achieved in 94.5% and subsidence occurred in 8.1% by the radiologic assessment. Statistical analysis reveals that the subsidence seen later than 8 weeks after surgery and the development of subsidence does not correlate statistically with the change of the postoperative disc space height. PMID- 19137082 TI - Effect of single growth factor and growth factor combinations on differentiation of neural stem cells. AB - OBJECTIVE: The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF), insulin growth factor-I (IGF-I), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. METHODS: Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using beta-tubulin III and NeuN as neural markers. RESULTS: Neural differentiations in the presence of individual growth factors for beta-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF 1, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for beta-tubulin III and 3.1% for NeuN. For beta tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1%; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). CONCLUSION: Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and postmitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation. PMID- 19137083 TI - Gas-forming brain abscess caused by Klebsiella pneumoniae. AB - Gas forming brain abscess is a rare disease caused by Klebsiella pneumoniae occurring in patients with impaired host defense mechanism such as diabetes mellitus or liver cirrhosis. A 59-year-old man with 2-year history of diabetes mellitus and 20-year history of liver cirrhosis presented to the hospital with headache. On the day after admission, severe headache was developed and he deteriorated rapidly. Brain CT showed a non-enhanced mass including multiple air density as well as surrounding edema seen in the right occipital lobe, and isodensity air-fluid level seen in the right lateral ventricle. Despite emergent ventricular drainage and intraventricular and intravenous administration of antibiotics, his condition progressively worsened to sepsis and to death after 5 days. Bacterial culture of blood and ventricular fluids disclosed a Gram (-) rod, Klebsiella pneumoniae. In this report we review the pathogenic mechanism and its management. PMID- 19137084 TI - Lumbar periradicular abscess mimicking a fragmented lumbar disc herniation : an unusual case. AB - We herein describe the case of a focal spontaneous spinal epidural abscess who was initially diagnosed to have a free fragment of a lumbar disc. A 71-year-old woman presented with history of low back and right leg pain. Magnetic resonance imaging suggested a peripherally enhancing free fragment extending down from S1 nerve root axilla. Preoperative laboratory investigation showed elevation of c reactive protein (CRP), erythrocyte sedimentation rate (ESR) levels. She was taken for surgery and a fluctuating mass at the axilla of S1 nerve was found. When the mass was probed with a dissector, a dark yellow, thick pus drained out. Pus cultures were negative. Patients who present with extreme low back plus leg pain and increased leucocyte count, ESR and CRP levels should raise the suspicion of an infection of a vertebral body or spinal epidural space. PMID- 19137085 TI - Congenital absence of a cervical spine pedicle : report of two cases and review of the literature. AB - Congenital absence of a cervical spine pedicle is a rare clinical entity, and it is usually found incidentally on radiological studies performed after trauma in patients with cervical pain. We report two cases of congenital absence of a cervical spine pedicle and present a review of the literature. PMID- 19137086 TI - Prone position-related meralgia paresthetica after lumbar spinal surgery : a case report and review of the literature. AB - Lateral femoral cutaneous neuropathy occurring during spinal surgery is frequently related to iliac bone graft harvesting, but meralgia paresthetica (MP) can result from the patient being in the prone position. Prone position-related MP is not an uncommon complication after posterior spine surgery but there are only few reports in the literature on this subject. It is usually overlooked because of its mild symptoms and self-limiting course, or patients and physicians may misunderstand the persistence of lower extremity symptoms in the early postoperative period to be a reflection of poor surgical outcome. The authors report a case of prone position-related MP after posterior lumbar interbody fusion at the L3-4 and reviewed the literature with discussion on the incidence, pathogenesis, and possible risk factors related to this entity. PMID- 19137087 TI - Delayed bilateral abducens nerve palsy after head trauma. AB - Although the incidence of unilateral abducens nerve palsy has been reported to be as high as 1% to 2.7% of head trauma cases, bilateral abducens nerve palsy following trauma is extremely rare. In this report, we present the case of a patient who developed a bilateral abducens nerve palsy and hypoglossal nerve palsy 3 days after suffering head trauma. He had a Glasgow Coma Score (GCS) of 15 points. Computed tomography (CT) images demonstrated clivus epidural hematoma and subarachnoid hemorrhage on the basal cistern. Herein, we discuss the possible mechanisms of these nerve palsies and its management. PMID- 19137088 TI - Hemifacial spasm caused by fusiform aneurysm at vertebral artery-posterior inferior cerebellar artery junction. AB - Hemifacial spasm induced by intracranial aneurysm is a rare clinical condition. A 45-year-old male patient presented with a 3-year history of progressive involuntary twitching movement on right face. On radiological study, a dilated vascular lesion compressing the brain stem was found at the junction of vertebral artery and posterior inferior cerebellar artery. On operative field, we found the posterior inferior cerebellar artery and the fusiform aneurysm compressing root exit zone of facial nerve. Microvascular decompression was performed and the facial symptom was relieved without complications. PMID- 19137089 TI - Primary intracranial squamous cell carcinoma in the brain stem with a cerebellopontine angle epidermoid cyst. AB - Primary intracranial squamous cell carcinoma is extremely rare, with most cases arising from a preexisting benign epidermoid cyst. We report a rare case of primary intracranial squamous cell carcinoma in the brain stem with a cerebellopontine angle (CPA) epidermoid cyst. A 72-year-old female suffered from progressive left hemiparesis, difficulty in swallowing, and right hemifacial numbness. Diffusion-weighted magnetic resonance imaging revealed a high signal intensity (SI) lesion in the CPA region and an intra-axially ring-enhanced cystic mass in the right brain stem with low SI. Whole-body positron emission tomography showed no evidence of metastatic disease. The histological findings revealed a typical epidermoid cyst in the CPA region and a squamous cell carcinoma in the brain stem. We speculate that the squamous cell carcinoma may have been developed due to a chronic inflammatory response by the adjacent epidermoid cyst. The patient underwent a surgical resection and radiotherapy. After 12 months, she had no evidence of recurrence. PMID- 19137090 TI - Intraventricular glioblastoma multiforme with previous history of intracerebral hemorrhage : a case report. AB - GBM is the most common primary brain tumor, but intraventricular GBM is rare and only few cases have been reported in the literature. The authors report a case of 64-year-old man who had a remote history of previous periventricular intracerebral hemorrhage. Brain computed tomography (CT) and magnetic resonance (MR) imaging showed an intraventricular lesion with inhomogeneous enhancement, infiltrative borders and necrotic cyst, and obstructive hydrocephalus. The patient underwent surgical removal through transcortical route via the bottom of previous hemorrhage site and the final pathologic diagnosis was GBM. We present a rare case of an intraventricular GBM with detailed clinical course, radiological findings, and pathological findings, and the possible origin of this lesion is discussed. PMID- 19137091 TI - Comparing techniques for the identification of the MTHFR A1298C polymorphism. AB - The restriction fragment length polymorphism (RFLP) technique with the MboII enzyme is used by a number of researchers as a methodology for the identification of the genetic polymorphism MTHFR A1298C. However, the reliability of this enzyme for genotyping this polymorphism has been questioned, since the silent polymorphism T1317C, located close to the polymorphic region A1298C on gene MTHFR, also has a recognition site for MboII. Thus, the fragments formed by the digestion of MboII present similar sizes, making it difficult to differentiate the allele MTHFR 1298A in the presence of the allele MTHFR 1317C. Hence, we investigated the A1298C polymorphism in a Brazilian population of renal transplant patients, using the RFLP technique with digestion by Mbo II and using sequencing, in order to examine the concordance between the two techniques. Our results showed an 8.6% difference in genotyping between RFLP and sequencing, but the statistical concordance test presented a kappa coefficient equal to 0.81 (CI 95% 0.74-88), which indicates a virtually perfect concordance, according to the criterion of Landis and Koch. Therefore, we concluded that the RFLP technique is concordant with automated sequencing in the detection of polymorphism A1298C under our laboratory conditions. PMID- 19137092 TI - Immobilized metal affinity electrophoresis: a novel method of capturing phosphoproteins by electrophoresis. AB - An immobilized metal affinity electrophoresis (IMAEP) method is described here. In this method, metal ions are immobilized in a native polyacrylamide gel to capture phosphoproteins. The capture of phosphoproteins by IMAEP is demonstrated with immobilized metals like iron, aluminum, manganese, or titanium. In the case studies, phosphoproteins alpha-casein, beta-casein, and phosvitin are successfully extracted from a protein mixture by IMAEP. PMID- 19137093 TI - Development of DNA aptamers to a foot-and-mouth disease peptide for competitive FRET-based detection. AB - We sought to develop a novel competitive fluorescence resonance energy transfer (FRET)-aptamer-based strategy for detection of foot-and-mouth (FMD) disease within minutes. A 14-amino-acid peptide from the VP1 structural protein, which is conserved among 16 strains of O-serotype FMD virus, was synthesized and labeled with Black Hole Quencher-2 (BHQ-2) dye. Polyclonal FMD DNA aptamers were labeled with Alexa Fluor 546-14-dUTP by polymerase chain reaction and allowed to bind the BHQ-2-peptide conjugate. Following purification of the FRET-aptamer-peptide complex, a "lights off" response was observed within 10 minutes and was sensitive to a level of 25-250 ng/mL of FMD peptide. Ten candidate aptamers were sequenced from the polyclonal family. The aptamer candidates were screened in an enzyme based plate assay. A high- and low-affinity aptamer candidate were each labeled with Alexa Fluor 546-14-dUTP by asymmetric polymerase chain reaction and used in the competitive FRET assay, but neither matched the sensitivity of the polyclonal FRET response, indicating the need for further screening of the aptamer library. PMID- 19137094 TI - LC-MALDI-TOF MS-based rapid identification of phenolic acids. AB - This study is the first on combined HPLC and MALDI-TOF MS analysis of phenolic acids. The analyses were carried out for phenolic acid mixtures and showed a unique, individual co-crystalline pattern for each phenolic acid. HPLC could distinguish phenolic acids and MALDI-TOF MS provided comparable mass (m/z) profiles for the samples. This combined study proved to be rapid in the accurate identification and structural analysis of phenolic acids with different masses. PMID- 19137095 TI - Assessing the quality of hybridized RNA in Affymetrix GeneChips using linear regression. AB - The quality of data from microarray analysis is highly dependent on the quality of RNA. Because of the lability of RNA, steps involved in tissue sampling, RNA purification, and RNA storage are known to potentially lead to the degradation of RNAs; therefore, assessment of RNA quality and integrity is essential. Existing methods for estimating the quality of RNA hybridized to a GeneChip either suffer from subjectivity or are inefficient in performance. To overcome these drawbacks, we propose a linear regression method for assessing RNA quality for a hybridized Genechip. In particular, our approach used the probe intensities from the .cel files that the Affymetrix software associates with each microarray. The effectiveness and the improvements of the proposed method over the existing methods are illustrated by the application of the method to the previously published 19 human Affymetrix microarray data sets for which external verification of RNA quality is available. PMID- 19137096 TI - Improved identification of membrane proteins by MALDI-TOF MS/MS using vacuum sublimated matrix spots on an ultraphobic chip surface. AB - Integral membrane proteins are notoriously difficult to identify and analyze by mass spectrometry because of their low abundance and limited number of trypsin cleavage sites. Our strategy to address this problem is based on a novel technology for MALDI-MS peptide sample preparation that increases the success rate of membrane protein identification by increasing the sensitivity of the MALDI-TOF system. For this, we used sample plates with predeposited matrix spots of CHCA crystals prepared by vacuum sublimation onto an extremely low wettable (ultraphobic) surface. In experiments using standard peptides, an up to 10-fold gain of sensitivity was found for on-chip preparations compared with classical dried-droplet preparations on a steel target. In order to assess the performance of the chips with membrane proteins, three model proteins (bacteriorhodopsin, subunit IV(a) of ATP synthase, and the cp47 subunit from photosystem II) were analyzed. To mimic realistic analysis conditions, purified proteins were separated by SDS-PAGE and digested with trypsin. The digest MALDI samples were prepared either by dried-droplet technique on steel plates using CHCA as matrix, or applied directly onto the matrix spots of the chip surface. Significantly higher signal-to-noise ratios were observed for all of the spectra resulting from on-chip preparations of different peptides.In a second series of experiments, the membrane proteome of Rhodococcus jostii RHA1 was investigated by AIEC/SDS-PAGE in combination with MALDI-TOF MS/MS. As in the first experiments, Coomassie-stained SDS-PAGE bands were digested and the two different preparation methods were compared. For preparations on the Mass.Spec.Turbo Chip, 43 of 60 proteins were identified, whereas only 30 proteins were reliably identified after classical sample preparation. Comparison of the obtained Mascot scores, which reflect the confidence level of the protein identifications, revealed that for 70% of the identified proteins, higher scores were obtained by on-chip sample preparation. Typically, this gain was a consequence of higher sequence coverage due to increased sensitivity. PMID- 19137100 TI - Highlights of Association for Biochemistry and Molecular Biology meeting during Experimental Biology 2008. PMID- 19137101 TI - Quartz crystal microbalance with dissipation monitoring: enabling real-time characterization of biological materials and their interactions. AB - In recent years, there has been a rapid growth in the number of scientific reports in which the quartz crystal microbalance (QCM) technique has played a key role in elucidating various aspects of biological materials and their interactions. This article illustrates some key advances in the development of a special variation of this technique called quartz crystal microbalance with dissipation monitoring (QCM-D). The main feature and advantage of QCM-D, compared with the conventional QCM, is that it in addition to measuring changes in resonant frequency (Deltaf), a simultaneous parameter related to the energy loss or dissipation (DeltaD) of the system is also measured. Deltaf essentially measures changes in the mass attached to the sensor surface, while DeltaD measures properties related to the viscoelastic properties of the adlayer. Thus, QCM-D measures two totally independent properties of the adlayer. The focus of this review is an overview of the QCM-D technology and highlights of recent applications. Specifically, recent applications dealing with DNA, proteins, lipids, and cells will be detailed. This is not intended as a comprehensive review of all possible applications of the QCM-D technology, but rather a glimpse into a few highlighted application areas in the biomolecular field that were published in 2007. PMID- 19137102 TI - Using metabolomics to estimate unintended effects in transgenic crop plants: problems, promises, and opportunities. AB - Transgenic crops are widespread in some countries and sectors of the agro economy, but are also highly contentious. Proponents of transgenic crop improvement often cite the "substantial equivalence" of transgenic crops to the their nontransgenic parents and sibling varieties. Opponents of transgenic crop improvement dismiss the substantial equivalence standard as being without statistical basis and emphasize the possible unintended effects to food quality and composition due to genetic transformation. Systems biology approaches should help consumers, regulators, and other stakeholders make better decisions regarding transgenic crop improvement by characterizing the composition of conventional and transgenically improved crop species and products. In particular, metabolomic profiling via mass spectrometry and nuclear magnetic resonance can make broad and deep assessments of food quality and content. The metabolome observed in a transgenic variety can then be assessed relative to the consumer and regulator accepted phenotypic range observed among conventional varieties. I briefly discuss both targeted (closed architecture) and nontargeted (open architecture) metabolomics with respect to the transgenic crop debate and highlight several challenges to the field. While most experimental examples come from tomato (Solanum lycoperiscum), analytical methods from all of systems biology are discussed. PMID- 19137103 TI - Population studies of intact vitamin D binding protein by affinity capture ESI TOF-MS. AB - Blood plasma proteins with molecular weights greater than approximately 30 kDa are refractory to comprehensive, high-throughput qualitative characterization of microheterogeneity across human populations. Analytical techniques for obtaining high mass resolution for targeted, intact protein characterization and, separately, high sample throughput exist, but efficient means of coupling these assay characteristics remain rather limited. This article discusses the impetus for analyzing intact proteins in a targeted manner across populations and describes the methodology required to couple mass spectrometric immunoassay with electrospray ionization mass spectrometry for the purpose of qualitatively characterizing a prototypical large plasma protein, vitamin D binding protein, across populations. PMID- 19137104 TI - Selective glutaraldehyde-mediated coupling of proteins to the 3'-adenine terminus of polymerase chain reaction products. AB - Attachment of proteins to the 3' end of DNA increases stability of the DNA in serum and retards clearance of DNA by major organs, thereby enhancing in vivo half-life and therapeutic potential of DNA. Unfortunately, the length of DNA molecules that can be produced with 3 ' modifications by solid-phase synthesis for protein attachment is limited to 45-60 nucleotides due to uncertainties about sequence fidelity for longer oligonucleotides. Here we describe selective covalent coupling of proteins or other molecules to the 3'-adenine overhang of unlabeled and fluorophore-labeled double-stranded polymerase chain reaction products putatively at the N6 position of adenine using 2.5% glutaraldehyde at pH 6.0 and 4 degrees C for at least 16 h. Gel mobility shift analyses and fluorescence analyses of the shifted bands supported conjugate formation between double-stranded polymerase chain reaction products and beta2-microglobulin. In addition, blunt-ended DNA ladder fragments treated with glutaraldehyde at 4 degrees C showed no evidence of DNA-DNA or DNA-protein conjugate formation. With the present cold glutaraldehyde technique, longer DNA-3'-protein conjugates might be easily mass-produced. The protein portion of a DNA-3'-protein conjugate could possess functionality as well, such as receptor binding for cell entry, cytotoxicity, or opsonization. PMID- 19137105 TI - Formation of template-switching artifacts by linear amplification. AB - Linear amplification is a method of synthesizing single-stranded DNA from either a single-stranded DNA or one strand of a double-stranded DNA. In this protocol, molecules of a single primer DNA are extended by multiple rounds of DNA synthesis at high temperature using thermostable DNA polymerases. Although linear amplification generates the intended full-length single-stranded product, it is more efficient over single-stranded templates than double-stranded templates. We analyzed linear amplification over single- or double-stranded mouse H-ras DNA (exon 1-2 region). The single-stranded H-ras template yielded only the intended product. However, when the double-stranded template was used, additional artifact products were observed. Increasing the concentration of the double-stranded template produced relatively higher amounts of these artifact products. One of the artifact DNA bands could be mapped and analyzed by sequencing. It contained three template-switching products. These DNAs were formed by incomplete DNA strand extension over the template strand, followed by switching to the complementary strand at a specific Ade nucleotide within a putative hairpin sequence, from which DNA synthesis continued over the complementary strand. PMID- 19137106 TI - Tissue fractionation by hydrostatic pressure cycling technology: the unified sample preparation technique for systems biology studies. AB - Major bottlenecks in systems biology studies arise from limitations of current sample preparation techniques. Multiple mutually exclusive sample preparation methods, which are often required to extract distinct classes of molecules from cells and tissues, are incompatible with studies of precious or very limited samples. Moreover, the strong detergents and chaotropic agents commonly required to solubilize sample constituents often interfere with subsequent separation and analysis. Here we describe a rapid, detergent-free sample preparation technique that allows efficient concurrent isolation and fractionation of protein, DNA, RNA, and lipids from biological samples, eliminating the need for multiple replicates. The method relies on a synergistic combination of physical disruption of the cellular material by hydrostatic pressure (pressure cycling technology) and novel extraction conditions to dissolve and partition distinct classes of molecules into separate fractions. We demonstrate parallel recovery of proteins, lipids, and intact DNA and RNA, from animal cells and tissues, for proteomic, lipidomic, and genomic analyses. The protein extracts require minimal cleanup and are compatible with 1D and 2D PAGE, liquid chromatography coupled with tandem mass spectrometry, and Western blotting. The lipid fractions have been profiled by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry without further processing. The isolated DNA and RNA were shown to be intact by agarose gel visualization, and the presence of intact mRNA was confirmed by real time reverse transcription polymerase chain reaction. Analysis and comparison of samples extracted using this method and a more traditional extraction technique revealed several protein species preferentially extracted by the new method. PMID- 19137107 TI - Application of atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry for rapid identification of Neisseria species. AB - Atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry (AP-MALDI MS) was applied to develop a proteomics-based method to detect and identify Neisseria species. Heat-inactivated clinical isolate cell suspensions of Neisseria gonorrhoeae and strains belonging to five serogroups (A, B, C, W135, and Y) of Neisseria meningitidis were subjected to on-probe protein/peptide extraction and tryptic digestion followed by AP-MALDI tandem MS (MS/MS)-based proteomic analysis. Amino acid sequences derived from three protonated peptides with m/z values of 1743.8, 1894.8, and 1946.8 were identified by AP-MALDI MS/MS and MASCOT proteome database search analysis as belonging to neisserial acyl carrier protein, neisserial-conserved hypothetical protein, and neisserial putative DNA binding protein, respectively. These three peptide masses can thus be potential biomarkers for neisserial species identification by AP MALDI MS. PMID- 19137108 TI - High performance DNA purification using a novel ion exchange matrix. AB - Ion exchange chromatography has emerged as a reliable alternative to classic CsCl ethidium bromide gradients for isolating nucleic acids of the highest purity. A plasmid purification method based on a unique anion exchange membrane (IEXM) was developed for the production of superior quality plasmids. This method was simpler and more efficient than conventional bead-based methods. Plasmids were extracted from bacterial cells through alkaline lysis. The crude lysate was clarified by a sequential filtration device that not only removed cell debris but micellar aggregates as well. The clarified lysate was mixed with an extraction solution and loaded into a spin column containing IEXM. Binding, washing, and elution conditions were optimized to achieve efficient isolation of plasmids from the impurities. IEXM had an exceedingly high dynamic binding capacity, excellent selectivity, and a near 100% recovery for plasmids. The binding capacity for pUC19 was 2.93 mg/cm(3) of IEXM, which is several times greater than the values for conventional ion exchange beads. The superior selectivity of the method was reflected in the extremely low levels of endotoxin, and thus it is well-suited for critical applications in eukaryotic systems. PMID- 19137109 TI - Hyphenated tools for phospholipidomics. AB - The analysis of intact and underivatised lipids in body fluids as well as in cell and tissue extracts is of utmost importance in the field of early diagnosis. Therefore, fast, reliable, and automated analytical methods are needed to detect known as well as unknown species. The combination of solid phase extraction, high performance liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectroscopy are best at meeting this challenge. Herein, we show a workflow for the reliable analysis of individual components in phosphatidylethanolamine extracts. The limitations and advantages of the individual methods are discussed. PMID- 19137110 TI - The use of EGFR exon 19 and 21 unlabeled DNA probes to screen for activating mutations in non-small cell lung cancer. AB - Activating mutations in epidermal growth factor receptor-1 (EGFR) are found in 10 15% of Caucasian patients with non-small cell lung carcinoma (NSCLC). Approximately 90% of the mutations are deletions of several amino acids in exon 19 or point mutations in exon 21. Some studies suggest that these mutations identify patients that might benefit from targeted EGFR inhibitor therapy. DNA melting analysis of polymerase chain reaction products can screen for these mutations to identify this patient population. However, amplicon DNA melting analysis, although easily capable of detecting heterozygous mutations by heterodimer formation, becomes more difficult if mutations are homozygous or if the mutant allele is selectively amplified over wild type. Amplification of EGFR is common in NSCLC and this could compromise mutation detection by amplicon melting analysis. To overcome this potential limitation, we developed unlabeled, single-stranded DNA probes, complimentary to EGFR exon 19 and exon 21 where the common activating mutations occur. The unlabeled probes are incorporated into a standard polymerase chain reaction during the amplification of EGFR exons 19 and 21. The probe melting peak is easily distinguished from the amplicon melting peak, and probe melting is altered if mutations are present. This allows for easy identification of activating mutations even in homozygous or amplified states and is useful in the screening of NSCLC for the common EGFR activating mutations. PMID- 19137112 TI - Increased potency and longevity of gene silencing using validated Dicer substrates. AB - Chemically synthesized small interfering RNAs (siRNAs) are tools used for silencing the expression of a single gene. They are mainly employed in basic research applications, but may also have great potential in therapeutic applications. Longer double-stranded RNAs, such as Dicer-substrate 27mers, trigger gene silencing through the intrinsic RNAi pathway. The design of these Dicer-substrate 27mers has been optimized so they can be oriented by Dicer to consistently select the antisense (guide) strand after cleavage to shorter siRNAs, leading to predictable mRNA cleavage. In this paper we describe evidence that these Dicer-substrate 27mers produce more potent and sustained gene silencing for four genes when compared with synthetic 21mers that have the same guide-strand sequence. Furthermore, improved silencing by these 27mers is often more pronounced at lower concentrations. PMID- 19137113 TI - Multiplex ligation-dependent probe amplification analysis on capillary electrophoresis instruments for a rapid gene copy number study. AB - Annotated DNA samples that had been previously analyzed were tested using multiplex ligation-dependent probe amplification (MLPA) assays containing probes targeting BRCA1, BRCA2, and MMR (MLH1/MSH2 genes) and the 9p21 chromosomal region. MLPA polymerase chain reaction products were separated on a capillary electrophoresis platform, and the data were analyzed using GeneMapper v4.0 software (Applied Biosystems, Foster City, CA). After signal normalization, loci regions that had undergone deletions or duplications were identified using the GeneMapper Report Manager and verified using the DyeScale functionality. The results highlight an easy-to-use, optimal sample preparation and analysis workflow that can be used for both small- and large-scale studies. PMID- 19137114 TI - Influence of diet on the proteome of Drosophila melanogaster as assessed by two dimensional gel electrophoresis and capillary liquid chromatography-mass spectrometry: the hamburger effect revisited. AB - Proteomic biomarker discovery has been called into question. Diamandis hypothesized that seemingly trivial factors, such as eating a hamburger, may cause sufficient proteomic change as to confound proteomic differences. This has been termed the hamburger effect. Little is known about the variability of complex proteomes in response to the environment. Two methods-two-dimensional gel electrophoresis (2DGE) and capillary liquid chromatography-electrospray ionization time-of-flight mass spectrometry (LCMS)-were used to study the hamburger effect in two cross-sections of the soluble fruit fly proteome. 2DGE measured abundant proteins, whereas LCMS measured small proteins and peptides. Proteomic differences between males and females were first evaluated to assess the discriminatory capability of the methods. Likewise, wild-type and white-eyed flies were analyzed as a further demonstration that genetically based proteomic differences could be observed above the background analytical variation. Then dietary interventions were imposed. Ethanol was added to the diet of some populations without significant proteomic effect. However, after a 24-h fast, proteomic differences were found using LCMS but not 2DGE. Even so, only three of approximately 1000 molecular species were altered significantly, suggesting that the influence of even an extreme diet change produced only modest proteomic variability, and that much of the fruit fly proteome remains relatively constant in response to diet. These experiments suggest that proteomics can be a viable approach to biomarker discovery. PMID- 19137115 TI - ABRF-PRG05: de novo peptide sequence determination. AB - A common request of proteomics core facilities is protein identification. However, in some instances primary sequence information for the protein in question is not present in public databases. In other cases, the amino acid sequence of a protein may differ in some way from the sequence predicted from the gene sequence in a database as a result of gene mutation, gene splicing, and/or multiple posttranslational modifications. Thus, it may be necessary to determine the sequence of one or more peptides de novo in order to identify and/or adequately characterize the protein of interest. The primary goal of this study was to give participating laboratories an opportunity to evaluate their proficiency in sequencing unknown peptides that are not included in any published database. Samples containing 3-6 pmol each of five synthetic peptides with amino acid sequences that were not present in public databases were sent to 106 laboratories. One nonstandard amino acid was present in one of the peptides. From a comparison of the results obtained by different strategies, participating laboratories will be able to gauge their own capabilities and establish realistic expectations for the approaches that can be used for this determination. PMID- 19137116 TI - Molecular formula and METLIN Personal Metabolite Database matching applied to the identification of compounds generated by LC/TOF-MS. AB - In an effort to simplify and streamline compound identification from metabolomics data generated by liquid chromatography time-of-flight mass spectrometry, we have created software for constructing Personalized Metabolite Databases with content from over 15,000 compounds pulled from the public METLIN database (http://metlin.scripps.edu/). Moreover, we have added extra functionalities to the database that (a) permit the addition of user-defined retention times as an orthogonal searchable parameter to complement accurate mass data; and (b) allow interfacing to separate software, a Molecular Formula Generator (MFG), that facilitates reliable interpretation of any database matches from the accurate mass spectral data. To test the utility of this identification strategy, we added retention times to a subset of masses in this database, representing a mixture of 78 synthetic urine standards. The synthetic mixture was analyzed and screened against this METLIN urine database, resulting in 46 accurate mass and retention time matches. Human urine samples were subsequently analyzed under the same analytical conditions and screened against this database. A total of 1387 ions were detected in human urine; 16 of these ions matched both accurate mass and retention time parameters for the 78 urine standards in the database. Another 374 had only an accurate mass match to the database, with 163 of those masses also having the highest MFG score. Furthermore, MFG calculated a formula for a further 849 ions that had no match to the database. Taken together, these results suggest that the METLIN Personal Metabolite database and MFG software offer a robust strategy for confirming the formula of database matches. In the event of no database match, it also suggests possible formulas that may be helpful in interpreting the experimental results. PMID- 19137117 TI - Miniaturized GPCR signaling studies in 1536-well format. AB - G protein-coupled receptors (GPCRs) are involved in various physiological processes, such as behavior changes, mood alteration, and regulation of immune system activity. Thus, GPCRs are popular targets in drug screening, and a well designed assay can speed up the discovery of novel drug candidates. The Promega cAMP-Glo Assay is a homogenous bioluminescent assay to monitor changes in intracellular cyclic adenosine monophosphate (cAMP) concentrations in response to the effect of an agonist, antagonist, or test compound on GPCRs. Together with the Labcyte Echo 555 acoustic liquid handler and the Deerac Fluidics Equator HTS reagent dispenser, this setup can screen compounds in 96-, 384-, and 1536-well formats for their effects on GPCRs. Here, we describe our optimization of the cAMP-Glo assay in 1536-well format, validate the pharmacology, and assess the assay robustness for HTS. We have successfully demonstrated the use of the assay in primary screening applications of known agonist and antagonist compounds, and confirmed the primary hits via secondary screening. Implementing a high throughput miniaturized GPCR assay as demonstrated here allows effective screening for potential drug candidates. PMID- 19137119 TI - Lubiprostone--a novel treatment for irritable bowel syndrome with constipation. AB - Irritable bowel syndrome (IBS) is a chronic, highly prevalent gastrointestinal motility disorder characterized by abdominal discomfort/pain associated with altered bowel habits such as diarrhea or constipation or both. Current therapy for the constipation-predominant form (IBS-C) comprises fiber or osmotic or stimulant laxatives. However, these may exacerbate the condition or cause electrolyte disturbances. Lubiprostone is a novel selective chloride channel-2 activator that increases fluid secretion in the intestinal apical cell membrane, increasing gut motility and frequency of stool passage, and alleviating abdominal discomfort/pain. Lubiprostone has very low systemic bioavailability and cannot be quantitated in blood, but its active metabolite, M3, has been pharmacokinetically profiled. Lubiprostone reaches peak plasma concentrations within approximately 1 h and has a half-life of 0.9-1.4 h. Despite this short half-life, lubiprostone can be administered orally twice daily. Its efficacy in IBS-C has been demonstrated in two phase III studies; spontaneous bowel movement frequency increased and stool consistency improved, whereas straining, bloating and severity of constipation decreased. The beneficial effects continued for up to 4 weeks after cessation of lubiprostone. Lubiprostone was well tolerated in the long-term, with nausea and diarrhea being the commonest adverse events. Further studies are ongoing in opioid-induced bowel dysfunction. PMID- 19137120 TI - Milnacipran hydrochloride: its efficacy, safety and tolerability profile in fibromyalgia syndrome. AB - Milnacipran is a serotonin-norepinephrine reuptake inhibitor (SNRI), although its norepinephrine reuptake inhibition predominates. It has been marketed in Europe and Japan as an antidepressant for many years, but only recently has it been investigated as a treatment for fibromyalgia syndrome (FMS). Both open-label and double-blind, placebo-controlled trials have confirmed its efficacy in FMS, not only on the pain component but also on the fluctuating array of other symptoms such as sleep and cognitive disturbances and fatigue The phase III trials have employed comprehensive composite endpoints to more accurately capture the many complex domains of FMS. Milnacipran's efficacy was not limited to the shortterm, 12-week trials. It also demonstrated durability of response for up to 1 year. Its safety and tolerability were typical of its class, and it was generally well tolerated. Further work will be required to establish its place in FMS therapy by comparing it directly with other agents. PMID- 19137121 TI - Dextromethorphan/quinidine sulfate for pseudobulbar affect. AB - A new agent containing a combination of dextromethorphan (DM) and quinidine (Q) is currently under development for the treatment of pseudobulbar affect (PBA). PBA is a disorder of emotional regulation, characterized by uncontrollable outbursts of laughing and/or crying that are disproportionate to the emotions being experienced. The pathophysiology of PBA is currently unknown, although the disorder is thought to occur exclusively in the setting of neurological disease. The most influential theory on PBA posits that emotional outbursts are being generated autonomously in the brain stem due to loss of regulatory control by the frontal lobe. Although rarely life-threatening, PBA can have significant impact on patient quality of life, and thus merits treatment. There are currently no approved treatments for PBA. Several agents have been found to be effective in small placebo-controlled trials and case series, with the most commonly used agents being tricyclic antidepressants and selective serotonin reuptake inhibitors. Both these treatments are inexpensive and relatively low-risk, although the quality and quantity of data available on their efficacy are not optimal. DM has several pharmacological mechanisms of action relevant to the brain. It is an N-methyl-D-aspartate (NMDA) receptor antagonist, which prompted investigators to study its potential for slowing progression in amyotrophic lateral sclerosis (ALS), where glutamate toxicity is thought to be a factor. The combination agent DM/Q was developed to slow the metabolism of DM by P450 2D6 enzymes in the liver. DM/Q was not effective in slowing ALS progression, but patients noted that it helped to control their emotional outbursts, suggesting it might be useful as a treatment for PBA. DM is also a sigma-1 receptor agonist. These receptors are widely distributed in the brain, but probably most heavily in the limbic system, suggesting that DM may exert its emotion-controlling effects via these receptors. The endogenous ligands for sigma-1 receptors are not altogether known, although they appear to include gonadal steroids. DM/Q was recently shown to be effective in reducing the severity of PBA in two large studies of ALS and multiple sclerosis, which are probably the most common neurological settings. These are the largest treatment studies of PBA ever done. The agent was safe and relatively well tolerated. Further studies are being conducted to see if efficacy can be maintained with lower doses of quinidine. If DM/Q is approved by the U.S. Food and Drug Administration for treatment of PBA, it would be the first agent approved for this purpose. Currently, the antidepressants are probably the most attractive pharmacologic options for treatment of PBA. The choice of whether to use DM/Q in this setting will likely depend on individual patient factors as well as cost. PMID- 19137122 TI - Pemetrexed disodium for the treatment of NSCLC: an update. AB - Pemetrexed disodium is a multitargeted antifolate cytotoxic chemotherapy agent approved by the U.S. Food and Drug Administration (FDA) initially for the treatment of malignant pleural mesothelioma, and in August 2004 for second-line treatment of non-small cell lung cancer (NSCLC). In September 2008, the FDA also approved pemetrexed and cisplatin as first-line therapy for NSCLC. Pemetrexed is also no longer recommended for treatment of NSCLC with squamous cell carcinoma histology. Pemetrexed is currently being tested in clinical trials as part of second-line combination, first-line, adjuvant and maintenance therapies. PMID- 19137123 TI - Fulvestrant--a novel estrogen receptor antagonist for the treatment of advanced breast cancer. AB - Endocrine therapy is often the preferred treatment option for postmenopausal women with hormone receptor-positive breast cancer. However, as many patients eventually develop resistance there is a need for novel, efficacious and well tolerated treatments that lack cross-resistance with current therapies. This review describes the development of fulvestrant (Faslodex), an estrogen receptor antagonist with a different and distinct mode of action and no agonist effects. Phase III clinical trials in postmenopausal women with advanced breast cancer have found fulvestrant at the approved dose of 250 mg/month to be at least as effective and well tolerated as anastrozole following disease progression or recurrence on tamoxifen, and as effective as exemestane following disease progression or recurrence on nonsteroidal aromatase inhibitors. In addition, fulvestrant has also demonstrated activity in patients with visceral and HER2+ disease, who are generally regarded as being less responsive to endocrine therapy. Data from a recent neoadjuvant study suggest that a higher dose of fulvestrant may possess greater activity. Trials evaluating higher fulvestrant doses, and regimens combining fulvestrant with aromatase inhibitors or agents targeting growth factor receptors, are ongoing and will help determine the optimum use of fulvestrant in the endocrine treatment sequence for postmenopausal women with advanced breast cancer. PMID- 19137124 TI - Celecoxib, NSAIDs and the skeleton. AB - Treating acute and chronic musculoskeletal pain is essential for improving healing of traumatic injuries and surgical procedures, and for improving patient quality of life. Physicians are limited primarily to treating musculoskeletal pain with nonsteroidal antiinflammatory drugs (NSAIDs), cyclooxygenase type 2 (COX-2)-selective NSAIDs such as celecoxib, or narcotics. Patients often treat their pain with over-the-counter NSAIDs. Unlike narcotics that target the central nervous system to alleviate pain, NSAIDs inhibit cyclooxygenase activity within the central nervous system and at the peripheral pain site to prevent the conversion of arachidonic acid into prostaglandins. Thus, NSAID use can and does alter certain fundamental processes involved in the normal healing of injured tissues. Cyclooxygenase activity and prostaglandin signaling are critical regulators of normal skeletal metabolism and inflammation related to injury or disease. Since most people only use NSAIDs sporadically to treat pain, few data indicate that short-term or repeated occasional use of NSAIDs is deleterious to skeletal health. However, clinical data suggest that chronic use of celecoxib, may impair normal skeletal function leading to decreased bone mineral density in older male patients. Experimental studies also have documented the negative effects of NSAIDs on healing of skeletal tissues. Fracture healing and tendon-to bone healing appear to be particularly susceptible to inhibition by celecoxib. Limited retrospective clinical data tends to support the experimental data that COX-2 function is critical for normal bone healing. In contrast, NSAID use and perhaps COX-2-selective NSAID use may be beneficial for healing of other skeletal injuries. In particular, NSAID use does not appear to have a long-term negative effect on the ultimate healing of tendons and ligaments. Indeed, NSAID therapy may inhibit adhesion formation during tendon healing, which leads to a better functional recovery. Certainly, NSAID therapy following acetabular fractures, other hip fractures, or following hip replacement surgery is beneficial for reducing heterotopic ossification that can limit joint mobility. The effects of NSAID or celecoxib therapy on healing of other skeletal tissues is less clear. For instance, clinical data indicates that celecoxib therapy does not impair spinal fusion but experimental data indicates the opposite. Similarly, some reports suggest that NSAID therapy may prevent further erosion of cartilage in certain arthritic conditions while other reports indicate that NSAID treatment will exacerbate cartilage damage. The difference in effects caused by NSAID or celecoxib therapy likely relate to the role cyclooxygenase has in the biology of the injured tissue or its healing response. Differences in pharmacology between NSAIDs, treatment regimens, experimental models and potential off-target effects also may confuse many of these issues. It is clear, however, that cyclooxygenase activity is involved in the healing of many skeletal tissues, either directly or indirectly through modulation of the inflammatory response. Consequently, pharmacological manipulation of cyclooxygenase using NSAIDs or celecoxib can profoundly affect skeletal health. PMID- 19137125 TI - Atherosclerotic plaque reduction: blood pressure, dyslipidemia, atherothrombosis. AB - Coronary heart disease (CHD) is one of the leading causes of morbidity and mortality worldwide and modification of established cardiovascular risk factors has become increasingly important in order to minimize the impact of the disease. In response to the considerable debate regarding the optimal use of therapeutic strategies, a range of biomarkers has been incorporated into clinical development programs in order to ascertain the efficacy of novel agents. In addition, technological advances in imaging of the artery wall now enable the quantification of the extent of atherosclerotic plaque and allow monitoring of serial changes in response to therapeutics. The most sensitive imaging modality to date is intravascular ultrasound (IVUS) which can determine and quantitate atheroma burden and has provided insights regarding the pathophysiology of plaque formation and the potential impact of new antiatherosclerotic therapies. Furthermore, serial IVUS has demonstrated that intensive modification of a number of established risk factors such as lipid levels and blood pressure has a beneficial impact on the rate of disease progression in patients with established CHD. PMID- 19137126 TI - Duloxetine for the treatment of fibromyalgia. AB - The underlying cause of fibromyalgia is not known, although dysfunction of serotoninergic and noradrenergic neurotransmitters appears to play an important role in the condition. Duloxetine is a newer and better tolerated dual antidepressant that does not induce muscarinic, histaminergic or adrenergic adverse reactions, and at the same time modulates and enhances the endogenous descending system that inhibits nociception. Duloxetine reduces pain symptoms in depression and other diseases and conditions, including fibromyalgia. Over 90% of the observed effect on pain is due to a direct analgesic effect rather than an indirect antidepressant effect. In clinical trials, pain reduction with duloxetine was not associated with its antidepressant and anxiolytic effects in patients with fybromialgia. A meta-analysis of four randomized, double-blind, placebo-controlled studies of duloxetine in the treatment of fibromyalgia showed it to be significantly superior to placebo in providing pain relief, reducing fatigue and improving physical and mental performance. The results of safety studies indicate that duloxetine is safe and well tolerated. Adverse effects tend to be mild, appearing more often at the start of therapy and decreasing or disappearing over the course of continued treatment. PMID- 19137127 TI - Efficacy and safety of recombinant factor VIII products in patients with hemophilia A. AB - The introduction of recombinant factor VIII (rFVIII) clotting factor concentrates nearly 20 years ago represented a significant advance in the treatment of hemophilia A. The major advantage of rFVIII products compared with plasma-derived FVIII products is related to product safety, with rFVIII products virtually eliminating bloodborne pathogen transmission. The most challenging aspect of hemophilia A management today is the development of FVIII inhibitors; previously untreated patients are at the highest risk for inhibitor formation. Presented in this article are results of clinical trials in previously treated and untreated patients and postmarketing surveillance studies for the four commercially available rFVIII products (Recombinate, ReFacto, Kogenate FS/Kogenate Bayer and Advate). Recombinant FVIII therapies are highly efficacious when used ondemand and prophylactically, and they have excellent safety profiles; there have been no reports of viral- or prion-based disease transmission associated with rFVIII administration. The incidence rate of inhibitors in previously untreated patients ranges from 15% to approximately 30%. Because rFVIII concentrates have proven efficacy and safety profiles, a number of hemophilia treatment groups recommend rFVIII products as first-line therapy in the management of hemophilia A. PMID- 19137128 TI - Epiceram for the treatment of atopic dermatitis. AB - EpiCeram consists of a specific combination of ceramides, cholesterol and fatty acids that mimics those naturally found in the skin. EpiCeram was approved by the FDA in April 2006 for use as a nonsteroidal lipid barrier emulsion to manage symptoms of burning and itching associated with dry skin conditions such as atopic dermatitis, irritant contact dermatitis, radiation dermatitis and other dermatoses. EpiCeram has shown comparable efficacy to a mid-strength topical corticosteroid in a clinical study involving 113 children with moderate to severe atopic dermatitis. Unlike topical steroids and immunomodulators such as calcineurin inhibitors that have clinically well-recognized undesirable side effects and usage restrictions (including FDA black box warning for the latter), current data suggests that EpiCeram has a favorable safety profile. Additionally, EpiCeram does not appear to have substantial restrictions associated with its use, especially with regards to the duration of use or patient age, compared to the other classes of prescription products. PMID- 19137129 TI - Pathophysiology and therapeutic strategies in the management of stroke: an update. AB - Strokes, or brain attacks, which occur due to an interruption of blood flow in discrete regions of the brain, are a major cause of death. Stroke is the third leading cause of death and disability in developed nations. The complex pathophysiology of this medical problem includes excitotoxicity mechanisms, inflammatory pathways, oxidative damage and ionic imbalances. The final event is the ischemic neuronal death which results in an irreversible loss of the neuronal function. Recently, significant therapeutic advances have been made which include use of carotid endarterectomy, thrombolytics, anticoagulant therapy, antiplatelet agents, neuroprotective agents, and treating associated risk factors such as hypertension and hyperlipemia. However, in spite of the significant development in these areas of stroke research, none of the available therapeutic options are curative in that they can reduce the burden of stroke. This article briefly summarizes the status of advances made in order to understand the neurobiology of stroke and provide insight into the therapeutic strategies that could be developed for the effective management of ischemic stroke. PMID- 19137130 TI - Oxymorphone: a mature molecule with new life. AB - The undertreatment of acute and chronic pain continues to be a significant health concern in the U.S. Opioids are recommended for the treatment of acute or chronic pain of moderate to severe intensity that is not responsive to other pharmacologic agents, such as nonsteroidal antiinflammatory drugs. A high level of interindividual responses to the analgesic effects and side effects of opioids necessitates the availability of multiple treatment options. Extended-release and immediate-release oral formulations of oxymorphone hydrochloride were recently approved by the U.S. Food and Drug Administration and may provide new options for patients who have not achieved adequate and well-tolerated analgesia with their current opioid. This review provides an overview of the basic pharmacology (including pharmacokinetic and pharmacodynamic profiles), clinical efficacy and tolerability of both oral oxymorphone formulations. PMID- 19137131 TI - Drugs for the treatment of diabetes complications. Zycose: a new player in the field? AB - Zycose is a newly released (2006) combined medication containing folic acid (1 mg) benfotiamine (150 mg) and benzamine (850 mg), a proprietary blend of para aminobenzoic acid (PABA), vitamin E and alpha-lipoic acid (ALA). Zycose protects vascular, retinal and kidney function by improving cellular health and promoting peripheral nerve health in people with diabetes. Zycose's therapeutic benefit is believed to be due to the additive effects of its compounds on lowering homocysteine levels (folic acid), reducing the production of advanced glycation end products (benfotiamine), improving endothelial function (folic acid, benfotiamine, ALA), reducing oxidative stress (ALA, vitamin E) and reducing carbonyl stress (benzamine). The complex composition of Zycose allows the therapeutic intervention of several hyperglycemiamediated disorders. The compound consists mainly of vitamins, therefore explaining, in part, the good safety profile and reduced adverse effects. PMID- 19137133 TI - Workshop report-KNAPPE (European Union Sixth Framework Project) final meeting. PMID- 19137134 TI - Moving on from our first decade of environmental science publishing. PMID- 19137135 TI - JEM spotlight: applications of advanced nanomaterials for environmental monitoring. Foreword. PMID- 19137136 TI - JEM Spotlight: Applications of advanced nanomaterials for environmental monitoring. AB - Rapid progress of the nanotechnology and advanced nanomaterials production offers significant opportunities for a wide range of applications for detection and remediation of a broad range of environmental contaminants. The convergence of analytical techniques and nanotechnology provides attractive possibilities for development of miniaturized, rapid, ultrasensitive and inexpensive methods for in situ and field-based environmental monitoring devices. This review provides an overview of the various nanoparticles and nanostructures used for this purpose, their integration into functional analytical devices, applications as electrode materials and gas sensing nanoprobes, in biosensors and as capture probes in immunomagnetic separations. Relevant, specific examples of nanomaterials-based chemical and biological sensors with applications in environmental monitoring are discussed. PMID- 19137137 TI - Cooking rice in a high water to rice ratio reduces inorganic arsenic content. AB - Total arsenic and arsenic speciation was performed on different rice types (basmati, long-grain, polished ([white] and wholegrain [brown]) that had undergone various forms of cooking. The effect of rinse washing, low volume (2.5 : 1 water : rice) and high volume (6 : 1 water : rice) cooking, as well as steaming, were investigated. Rinse washing was effective at removing circa. 10% of the total and inorganic arsenic from basmati rice, but was less effective for other rice types. While steaming reduced total and inorganic arsenic rice content, it did not do so consistently across all rice types investigated. Low volume water cooking did not remove arsenic. High volume water : rice cooking did effectively remove both total and inorganic arsenic for the long-grain and basmati rice (parboiled was not investigated in high volume cooking water experiment), by 35% and 45% for total and inorganic arsenic content, respectively, compared to uncooked (raw) rice. To reduce arsenic content of cooked rice, specifically the inorganic component, rinse washing and high volume of cooking water are effective. PMID- 19137138 TI - Polynuclear aromatic hydrocarbons (PAHs) in global background soils. AB - The levels and distribution of polynuclear aromatic hydrocarbons (PAHs) were investigated in global background soil samples. Total PAH concentrations (sum of 15 compounds) ranged over 5 orders of magnitude, from <1 to 7,840 (mean 328) ng g(-1) dry weight. The order was generally Europe > North America > Asia > Oceania > Africa > South America. Proximity to long-term emissions sources and locations susceptible to high atmospheric depositional inputs tended to have higher concentrations. A broad positive correlation was obtained between population density and soil PAH concentrations (i.e. source-related factors). However, concentrations were also influenced by the holding capacity of the soils (i.e. sink-related factors) with statistically significant correlations observed between PAHs and soil organic matter (SOM), and black carbon (BC). PMID- 19137139 TI - Detection of organophosphate pesticides using a prototype liquid crystal monitor. AB - The precision and accuracy of a prototype wearable liquid crystal monitor (LCM) for the measurement of airborne organophosphate pesticide concentrations was explored in a series of laboratory experiments. LCM response to vapor-phase and aerosol diazinon was compared to concentrations obtained using a standard reference method (NIOSH 5600) at concentrations ranging from approximately 8 to 108 ppb (parts per billion) over durations of 2 to 80 hours. Temperature ( approximately 25, 30, and 35 degrees C) and relative humidity (15, 50, and 85%) were varied to estimate the effect of these factors on LCM performance. The LCM response to vapor phase pesticide exposure was linear for concentrations in the range of 8-20 ppb. At exposure concentrations above approximately 20 ppb, however, there was a decline in monitor response and measurement precision. Elevated temperatures improved diazinon vapor-only measurement precision, while increased relative humidity reduced LCM response at the extremes of tested temperatures. Compared to vapor-only exposures, the LCM was less sensitive to diazinon aerosol concentrations, but displayed reasonable precision over a relatively large range of exposures (29 to 1190 ppb-hr). Further efforts to characterize temperature and humidity effects and improve low-end sensitivity would likely provide a portable personal exposure monitor or environmental sensor for this widely used class of pesticides. PMID- 19137140 TI - Detachment of tobacco-smoke-material carriers from surfaces by turbulent air flow. AB - The influence of tobacco-smoke-material (TSM) on the detachment of microparticles from surfaces by turbulent air flow was investigated experimentally. Both clean and dusty glass surfaces were subjected to TSM either before or after the deposition of 64 microm to 76 microm-diameter stainless steel microparticles onto the surfaces. The TSM was generated by mechanically puffing research-grade cigarettes inside a smoking box that contained the surfaces. Microparticle detachment characteristics were studied in a wind tunnel using video microphotography. Measured nicotine concentration was used to determine the amount of TSM deposited on a surface.The 5% and 50% threshold velocities for detachment were used to quantify the effect of TSM on microparticle detachment. These velocities were compared with those obtained using a clean surface with no TSM exposure. The effect of TSM exposure on microparticle detachment depended significantly on whether exposure occurred before or after microparticle deposition. TSM exposure before microparticle deposition had little effect. TSM exposure after deposition delayed detachment to much higher velocities. The presence of dust on the surface with TSM also delayed detachment and increased the variability in the detachment velocities as compared to the case of a clean surface with no TSM exposure. PMID- 19137141 TI - Image analysis of epicuticular damage to foliage caused by dry deposition of the air pollutant nitric acid. AB - Nitric acid vapor is produced by the same photochemical processes that produce ozone. In the laboratory, concentrated nitric acid is a strong acid and a powerful oxidant. In the environment, where the concentrations are much lower, it is an innocuous source of plant nitrogen. As an air pollutant, which mode of action does dry deposition of nitric acid follow? We investigated the effects of dry deposition of nitric acid on the foliage of four tree species native to the western United States. A novel controlled environment, fumigation system enabled a four-week exposure at concentrations consistent with ambient diurnal patterns. Scanning electron microscopy and automated image analysis revealed changes in the epicuticular wax layer during fumigation. Exposure to nitric acid resulted in a reproducible suite of damage symptoms that increased with increasing dose. Each tree species tested exhibited a unique set of damage features, including cracks, lesions, and conformation changes to epicuticular crystallite structures. Dry deposition of atmospheric nitric acid caused substantial perturbation to the epicuticular surface of all four tree species investigated, consistent with the chemical oxidation of epicuticular waxes. Automated image analysis eliminated many biases that can trouble microscopy studies. Trade names and commercial enterprises or products are mentioned solely for information. No endorsements by the U.S. Department of Agriculture are implied. PMID- 19137142 TI - Foliar loading and metabolic assimilation of dry deposited nitric acid air pollutants by trees. AB - Dry deposition of nitric acid vapor (HNO(3)) is a major contributor to eutrophication of natural ecosystems. Although soil fertilization by nitrogen deposition is considered to be the primary pathway for changes in plant nutrient status and shifts in ecological structure, the aerial portion of plants offer many times the surface area in which to collect atmospheric HNO(3). As much as 60% of deposited nitrogen may be retained in the canopy and not land on the soil surface below. Although uptake and assimilation appears to contribute to retention, only a small percentage of dry deposition is recovered in assimilated N pools. To test the importance of biological activity on the process and measurements of dry deposition, we used controlled environmental chambers to compare deposition to living and freeze-dried foliage of four tree species using (15)N-labeled HNO(3). In living trees, assimilation was determined by (15)N incorporation into free amino acids and proteins in leaves and roots. From 10% to 60% of the retained HNO(3) was incorporated into the biologically active nitrogen pool. The remainder was bound to foliar surfaces in an insoluble form in either living or freeze-dried foliage. The importance of the boundary layer conditions emerged as a primary factor controlling dry deposition characteristics and measurements.Trade names and commercial enterprises or products are mentioned solely for information. No endorsements by the U.S. Department of Agriculture are implied. PMID- 19137143 TI - Polycyclic aromatic hydrocarbons, polychlorinated biphenyls, phthalates and organotins in northern Atlantic Spain's coastal marine sediments. AB - Surface sediment samples (0-10 cm) from ten shallow marine sediments affected by industry and shipping traffic on the northern Atlantic Spanish coast were analysed to determine prevailing concentrations of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), phthalates and organotins. PAHs were detected at eight of the study areas (1.4 to 4.9 microg g(-1)), while PCBs were detected only at four of the study areas (2.9 to 37 ng g(-1)). PAHs concentration rations indicated that PAHs were mainly of pyrogenic origin. PCB congener patterns in all of the sediment samples were the same, and contained the less volatile congeners PCB-138, PCB-153 and PCB-180. Bis(2-ethyl hexyl)phthalate was the most abundant phthalate (190 to 2,600 ng g(-1)). Total organotin concentrations varied widely from 7.7 to 489 ng g(-1). A significant correlation was found between PAH concentrations and sediment particle sizes (p<0.001). Peak concentrations of organotins have the potential to induce ecotoxicological impacts based on levels specified in international Sediment Quality Guidelines, although the majority of the stations analysed are included in the medium-low range of priority. PMID- 19137144 TI - Characterization of the planktonic shrimp, Acetes intermedius, as a potential biomonitor for butyltin. AB - Acute toxic responses as well as uptake and depuration rates for tributyltin (TBT) and dibutyltin (DBT) were examined in the small planktonic shrimp, Acetes intermedius. The 72-h LC(50) values of TBT and DBT for the shrimp were found to be 18.6 and 82.6 microg L(-1) as tin. The uptake rate constants of TBT and DBT in the shrimp were 0.0006 and 0.0002 L g(-1) h(-1), and the corresponding depuration rate constants were 0.0303 and 0.0106 h(-1), respectively. It appears that real time ambient TBT pollution status can be more closely reflected in this species. The shrimp may serve as a biomonitor to indicate short-term fluctuations in ambient TBT pollution. A field survey was also conducted to distinguish contrasts in butyltin accumulation under different ambient conditions. These observations provide valuable information for the evaluation of TBT pollution status in the environment using A. intermedius as a biomonitor. PMID- 19137145 TI - Temporal evolution of imposex in Nassarius reticulatus (L.) along the Portuguese coast: the efficacy of EC regulation 782/2003. AB - Imposex levels in Nassarius reticulatus (L.) were determined in 44 sites along the Portuguese coast in 2006 in order to describe spatial and temporal trends of TBT pollution in the area. The percentage of females with imposex across sites varied between 20 and 100, denoting the extent of this phenomenon throughout the Portuguese coast. The mean female penis length per site varied between 0.0-8.0 mm and the relative penis length index (mean female penis length x 100/mean male penis length) attained a maximum value of 92%, i.e., female penis never surpasses the size of the male penis but nevertheless it can almost approach the male dimensions. The vas deferens sequence index ranged from 0.2 to 4.5 and the oviduct convolution index varied between 0.0 and 1.3 across stations. The penis growth, the vas deferens development and the oviduct convolution were all correlated and constitute visible signs of a global virilisation progression in females in response to the proximity of harbours that constitute the main TBT pollution sources. The results indicate that about 95% of the surveyed sites were still exposed to TBT water concentrations above the OSPAR Environmental Assessment Criteria. Nevertheless, signs of recovery are shown by the significant reduction of VDSI levels in 2006 in comparison to 2003, which points to the efficacy of the EC Regulation 782/2003 in reducing TBT pollution levels in the Portuguese coast. PMID- 19137147 TI - Analysis of 50-y record of surface (137)Cs concentrations in the global ocean using the HAM-global database. AB - We investigated spatial and temporal variations in (137)Cs concentrations in the surface waters of the global ocean for the period from 1957 to 2005 using the "HAM database - a global version". Based on the 0.5-y average value of (137)Cs concentrations in the surface water in each sea area, we classified the temporal variations into four types. (1) In the North Pacific Ocean where there was high fallout from atmospheric nuclear weapons tests, the rates of decrease in the (137)Cs concentrations changed over the five decades: the rate of decrease from the 1950s to the 1970s was much faster than that after the 1970s, and the (137)Cs concentrations were almost constant after the 1990s. Latitudinal differences in (137)Cs concentrations in the North Pacific Ocean became small with time. (2) In the equatorial Pacific and Indian Oceans, the (137)Cs concentrations varied within a constant range in the 1970s and 1980s, suggesting the advection of (137)Cs from areas of high global fallout in the mid-latitudes of the North Pacific Ocean. (3) In the eastern South Pacific and Atlantic Oceans (south of 40 degrees S), the concentrations decreased exponentially over the five decades. (4) In the Arctic and North Atlantic Oceans, including marginal seas, (137)Cs concentrations were strongly controlled by discharge from nuclear reprocessing plants after the late 1970s. The apparent half-residence times of (137)Cs in the surface waters of the global ocean from 1970 to 2005 ranged from 4.5 to 36.8 years. The apparent half-residence times were longer in the equatorial region and shorter in the higher latitudes. There was no notable difference between the latitudinal distributions of the apparent half-residence times in the Pacific and Indian Oceans. These results suggest that (137)Cs in the North Pacific Ocean is transported to the equatorial, South Pacific, and Indian Oceans by the oceanic circulation. PMID- 19137146 TI - Investigation of the matrix effects on a HPLC-ESI-MS/MS method and application for monitoring triazine, phenylurea and chloroacetanilide concentrations in fresh and estuarine waters. AB - In this work, the effects of matrix interferences on the analytical performance of a new multiresidue method based on off-line solid phase extraction followed by reversed-phase liquid chromatographic separation and electrospray triple quadrupole mass spectrometric detection were investigated. This technique allows the simultaneous determination of 30 triazines, phenylureas and chloroacetanilides, extracted from freshwaters, in 40 minutes. Quantifications were performed with the use of appropriate internal standards (i.e. atrazine D5, diuron D6 and metolachlor D6). The limits of quantification were from 1 to 32 ng L(-1) for the triazines, from 5 to 59 ng L(-1) for the phenylureas and from 13 to 54 ng L(-1) for the chloroacetanilides. The matrix effects were studied by spiking various waters (i.e. tap, river, pond and sea waters) with the chemicals of interest. The results showed that the samples with the highest conductivity (i.e. seawater) and the most abundant dissolved organic matter content (i.e. pond water) exhibited important matrix effects with signal suppressions and high imprecision, respectively. These matrix effects were strongly minimized by performing appropriate internal standardizations. Afterward, this analytical method was applied for analyzing environmental samples from either river or estuarine waters and for monitoring herbicide input in a freshwater-seawater interface. PMID- 19137148 TI - Diversity of hepatotoxic cyanobacteria in the Nile Delta, Egypt. AB - Optimized denaturing gradient gel electrophoresis (DGGE) fingerprinting and real time PCR were performed to identify and quantify the potential hepatotoxic microcystin- and nodularin-producing cyanobacteria present in freshwater samples collected during different seasons at three different sites from the Nile River Delta. The combined use of molecular gene markers (targeting the aminotransferase domain of the hepatotoxin synthetase modules and the 16S rRNA gene) and light microscopy demonstrated the dominance of different freshwater Microcystis phylotypes, including the potential hepatotoxin producers M. wesenbergii and M. aeruginosa, several Synechococcus and Cyanobium phylotypes, as well as the presence of Nodularia spumigena and Cylindrospermopsis raciborskii in the samples "collected during summer 2006 and winter 2007". Certain seasonal changes (summer and winter) in Microcystis microdiversity were detected. Real-time PCR revealed no difference in the quantities of potential hepatotoxin-producing cyanobacterial communities between summer and winter, but did show differences between the three sampling sites investigated. The expression of the aminotransferase domain analyzed by DGGE fingerprinting demonstrated that all Microcystis phylotypes present in the samples might have been active at the time of the sampling. Statistical analysis showed a significant effect of TP, and not TN, on the relative abundance of the potentially hepatotoxic cyanobacterial community. PMID- 19137149 TI - Evaluation of a hybrid constructed wetland for treating domestic sewage from individual housing units surrounding agricultural villages in South Korea. AB - The treatment efficiency of 2- and 3-stage constructed wetlands (CWs) was evaluated for treating domestic sewage from houses surrounding agricultural villages. The optimum depth of filter media was 90 cm. The optimum volume ratio of vertical flow (VF) and horizontal flow (HF) beds was 1:2, and the optimum filter medium was broken stone in the VF-HF 2-stage hybrid CWs. Based on the above optimum conditions, removal efficiency of biochemical oxygen demand (BOD), chemical oxygen demand (COD), suspended solids (SS), total nitrogen (T-N), and total phosphorus (T-P) were 99, 98, 99, 68, and 72%, respectively. However to utilize constructed wetlands (CWs) for treating domestic sewage for an individual house, would require downsizing of the 2-stage hybrid CWs. In addition, the low removal efficiency of T-N and T-P in 2-stage hybrid CWs would require improvements necessary to meet acceptable water quality discharge standards. Thus, to reduce the CWs' area and improve the T-N and T-P removal efficiencies, VF-HF 2-stage hybrid CW was modified into VF/HF(I)-HF(II) and VF/HF(I)-HF(III) 3 stage hybrid CW. The optimum reduced size of 3-stage hybrid CW was the VF/HF(I) HF(II) configuration which also increased T-N removal. Using this system, removal efficiency of BOD, COD, SS, T-N, and T-P were 99, 98, 99, 83, and 75%, respectively. In VF/HF(I)-HF(II) CW, the removal velocity of BOD, COD and SS was rapid on the order of VF (1st stage) >> HF(II) (3rd stage) > or = HF(I) (2nd stage), VF (1st stage) >> HF(II) (3rd stage) > HF(I) (2nd stage) and VF (1st stage) >> HF(I) (2nd stage) > HF(II) (3rd stage), respectively. The removal velocity of T-N and T-P in VF/HF(I)-HF(II) CWs was rapid on the order of HF(I) (2nd stage) > HF(II) (3rd stage) > or = VF (1st stage) and VF (1st stage) > HF(I) (2nd stage) > HF(II) (3rd stage), respectively. PMID- 19137150 TI - Uptake, accumulation and metabolic response of ferricyanide in weeping willows. AB - The remediation potential and metabolic responses of plants to ferricyanide were investigated using pre-rooted weeping willows (Salix babylonica L.) grown hydroponically in growth chambers and treated with potassium ferricyanide. Positive responses were observed for the plants exposed to 95% for real-world paints, National Institute of Standards and Technology's standard reference materials, and audit samples from the American Industrial Hygiene Association's Environmental Lead Proficiency Analytical Testing Program. This quantitative extraction procedure, when paired with quantitative paint sample collection and lead determination, may enable the development of a lead paint test kit that will meet the specifications of the final EPA rule. PMID- 19137154 TI - Personal exposure to inhalable cement dust among construction workers. AB - Objective- A case study was carried out to assess cement dust exposure and its determinants among construction workers and for comparison among workers in cement and concrete production.Methods- Full-shift personal exposure measurements were performed and samples were analysed for inhalable dust and its cement content. Exposure variability was modelled with linear mixed models.Results- Inhalable dust concentrations at the construction site ranged from 0.05 to 34 mg/m(3), with a mean of 1.0 mg/m(3). Average concentration for inhalable cement dust was 0.3 mg/m(3) (GM; range 0.02-17 mg/m(3)). Levels in the ready-mix and pre cast concrete plants were on average 0.5 mg/m(3) (GM) for inhalable dust and 0.2 mg/m(3) (GM) for inhalable cement dust. Highest concentrations were measured in cement production, particularly during cleaning tasks (inhalable dust GM = 55 mg/m(3); inhalable cement dust GM = 33 mg/m(3)) at which point the workers wore personal protective equipment. Elemental measurements showed highest but very variable cement percentages in the cement plant and very low percentages during reinforcement work and pouring. Most likely other sources were contributing to dust concentrations, particularly at the construction site. Within job groups, temporal variability in exposure concentrations generally outweighed differences in average concentrations between workers. 'Using a broom', 'outdoor wind speed' and 'presence of rain' were overall the most influential factors affecting inhalable (cement) dust exposure.Conclusion- Job type appeared to be the main predictor of exposure to inhalable (cement) dust at the construction site. Inhalable dust concentrations in cement production plants, especially during cleaning tasks, are usually considerably higher than at the construction site. PMID- 19137155 TI - Mathematical form factor studies on the effect of water on airborne particles morphology using a bi-dimensional TEM image processing. AB - Mathematical morphology is a tool for extracting image components that are useful for representation and description. The technique consists of a set-theoretic method of image analysis providing a quantitative description of geometrical structures. A simple application of mathematical morphology to a bi-dimensional processing of TEM images of airborne particles allows us to distinguish between particles grown and/or transported in atmosphere under dry conditions or in rainy days by a simple comparison of the corresponding image form factors. The form factors range in the 0.385-0.031 interval in the case of particles sampled in rainy days, and in the 0.103-0.006 interval in the case of non-rainy conditions. The same classification criterion was applied to filters collected under dry conditions and plunged in water. The results demonstrate that a morphological change may be artificially induced to the particle structure. The artificially wet particles, indeed, display an apparent contraction of their structures evidenced by a two-fold increase of the average values of their form factors. The last experiment roughly simulates the impact of particles on membranes of the respiratory tract. PMID- 19137156 TI - Distribution and source of alkyl polycyclic aromatic hydrocarbons in dustfall in Shanghai, China: the effect on the coastal area. AB - Alkyl polycyclic aromatic hydrocarbons (APAHs) are a group of persistent organic pollutants which are widely distributed in the atmospheric environment. To estimate the seasonal and spatial distribution of APAHs in dustfall in Shanghai, supercritical fluid extraction (SFE) followed by gas chromatography/mass spectrometry (GC/MS) were the analytical methods applied to the dustfall samples collected in this area from Dec. 2004 to Oct. 2005. Forty-one APAHs were found and divided into six groups which were alkyl naphthalenes (ANAs), alkyl phenanthrenes (APHs), alkyl anthracenes (AANs), alkyl fluoranthenes (AFLs), alkyl pyrenes (APYs) and alkyl chrysenes (ACHs). ANAs and APHs were two dominant APAHs in the proportion of more than 50%. The concentrations of total APAHs ranged from 1.6 to 9.0 microg g(-1). The ratios of APAHs to TPAHs (PAHs plus APAHs) fell into the range of 28 approximately 50%. The APAH levels were found to be higher in winter and spring than in summer and autumn. In terms of spatial distribution, concentration of APAHs was found to be higher in western Shanghai than eastern Shanghai. The source analysis showed that dustfall APAHs mainly derived from vehicle emissions and used crankcase oil. The annual fluxes of APAHs and PAHs in dustfall in urban Shanghai were 0.53 approximately 2.97 T and 0.96 approximately 5.34 T, respectively. PMID- 19137157 TI - Assessment of a procedure to determine trace and major elements in atmospheric aerosol. AB - The determination of trace elements in atmospheric particulate is affected by a number of problems that arise from some critical points such as the blank of the filters, sample heterogeneity and pre-analytical treatments. In the framework of a monitoring campaign conducted in the Venice Lagoon the analytical methodology for the determination of 20 trace elements (Al, As, Ca, Cd, Co, Cr, Cs, Cu, Fe, K, Li, Mg, Mn, Na, Ni, Pb, Rb, Sr, V, Zn) in atmospheric particulate samples by inductively coupled plasma quadrupole mass spectroscopy (ICP-QMS) has been optimized taking into account the individual critical points. Tests were carried out to estimate the blank contributions, and minimize the detection limit (LOD), measurements were also carried out to evaluate the accuracy and the repeatability. To obtain a complete dissolution of aerosol dust material and good recoveries of the elements, the acid mixture and the microwave assisted digestion program were optimized. The blank contributions from membrane filter manipulation and transportation prior to exposure were tested for the slotted and back filters by placing them on the sampling device for some minutes without air flowing to obtain field blanks (FBs). The contribution to the blank values of passive deposition and by contact with the samplers (quoted as campaign blanks, CBs) was measured by exposing the membranes throughout the sampling session (fifteen days) without any air flow. Instrumental ICP-QMS parameters were optimized and calibration curve intervals were selected on the basis of the necessity of simultaneous determination of the elements present at different levels of concentration. The limits of detection for each elements and the investigated method were suitable to determine the 20 elements reported above in the atmospheric aerosol fractionated in 6 classes ranging between 10 to 0.49 microm. It allows the determination of trace elements in aerosol in a large range of concentrations that can be observed in areas characterized by remarkable variability and regions with different levels of contamination. PMID- 19137158 TI - Ambient air quality and drug aftermaths of the Notte Bianca (White Night) holidays in Rome. AB - The environmental impact of Notte Bianca (White Night) holidays held during September 2007 in Rome, Italy, was investigated by measuring aerial concentrations of several genotoxic, psychotropic or source-dependent organic pollutants. Chemical characterization of airborne particulates collected concurrently at Montelibretti RM (semi-rural locality lying approximately 30 km NE of Rome centre) was used as a reference. n-Alkanes, polynuclear aromatic hydrocarbons (PAH) and nitrogen/oxygen-containing polyaromatic compounds (PAC) did not seem to undergo important enhancements during the holiday celebration, whilst recreational drugs peaked downtown (87 vs 41 pg m(-3), on the average, of cocaine, and 5.0 vs. 2.6 ng m(-3) of nicotine). By contrast, a simultaneous raising of aerial drugs was not detected at Montelibretti. PMID- 19137159 TI - Evaluation of HEPA vacuum cleaning and dry steam cleaning in reducing levels of polycyclic aromatic hydrocarbons and house dust mite allergens in carpets. AB - Dry steam cleaning, which has gained recent attention as an effective method to reduce house dust mite (HDM) allergen concentration and loading in carpets, was evaluated in this study for its efficacy in lowering levels of polycyclic aromatic hydrocarbons (PAHs) as well as HDM allergens. Fifty urban homes with wall-to-wall carpets, mostly low-income and with known lead contamination, were studied in 2003 and 2004. Two carpet-cleaning interventions were compared: Repeated HEPA (High Efficiency Particulate Air filtered) vacuuming alone and repeated HEPA vacuuming supplemented with dry steam cleaning. Vacuum samples were collected to measure carpet loading of dust and contaminants immediately before and after cleaning. Paired comparisons were conducted to evaluate the effectiveness of the cleaning protocols in reducing the levels of PAHs and HDM allergens in carpets. The results indicated that both cleaning methods substantially reduced the loading of PAHs and HDM allergens as well as dust in carpets (p < 0.0001). The reductions in loading of dust (64.4%), PAHs (69.1%), and HDM allergens (85.5%), by dry steam cleaning plus repetitive HEPA vacuuming were larger than the reductions by regular HEPA vacuuming alone: dust (55.5%), PAHs (58.6%), and HDM allergens (80.8%), although the difference was statistically significant only for dust and PAHs. We conclude that intensive HEPA vacuum cleaning substantially reduced the loading of PAHs and HDM allergens in carpets in these urban homes and that dry steam cleaning added modestly to cleaning effectiveness. PMID- 19137160 TI - A novel method for the in situ calibration of flow effects on a phosphate passive sampler. AB - Monitoring of nutrients including phosphate in the aquatic environment remains a challenge. In the last decade passive sampling techniques have been developed that facilitates the time integrated monitoring of phosphate (P) through the use of an iron hydroxide (ferrihydrite) to sequester dissolved phosphate from solution. These methods rely on established techniques to negate the effects of flow (and associated turbulence) and control the rate at which chemicals accumulate within passive samplers. In this study we present a phosphate sampler within which a suspension of ferrihydrite is contained behind a commercially available membrane. Accumulation of dissolved phosphates into the P-sampler is governed by the rate at which ions are diffusing through the membrane and the water boundary layer (WBL). As the WBL changes subject to flow we have adopted an in situ calibration technique based on the dissolution of gypsum to predict the change in the rate of uptake dependent on flow. Here we demonstrate that the loss of gypsum from the passive flow monitor (PFM) can be used to predict the sampling rate (the volume of water extracted per day) for phosphate as a function of water velocity. The outcome of this study presents a new in-field tool for more accurate prediction of the effect of flow/turbulence on the uptake kinetics into passive samplers that is controlled by the diffusion of the chemical of interest through the stagnant water boundary layer. PMID- 19137161 TI - Field comparison of passive air samplers with reference monitors for ambient volatile organic compounds and nitrogen dioxide under week-long integrals. AB - This study evaluates performance of nitrogen dioxide (NO2) and volatile organic compound (VOC) passive samplers with corresponding reference monitors at two sites in the Detroit, Michigan area during the summer of 2005. Ogawa passive NO2 samplers and custom-made, re-useable Perkin-Elmer (PE) tubes with Carbopack X sorbent for VOCs were deployed under week-long sampling periods for six weeks. Precise results (5% relative standard deviation, RSD) were found for NO2 measurements from collocated Ogawa samplers. Reproducibility was also good for duplicate PE tubes for benzene, toluene, ethylbenzene, and xylene isomers (BTEX species, all < or = 6% RSD). As seen in previous studies, comparison of Ogawa NO2 samplers with reference chemiluminescence measurements suggested good agreement. Generally good agreement was also found between the PE tubes and reference methods for BTEX species. PMID- 19137162 TI - Determination of ammonium in marine waters using a gas diffusion multicommuted flow injection system with in-line prevention of metal hydroxides precipitation. AB - A multi-commuted flow system coupled to a gas diffusion device was developed for the spectrophotometric determination of ammonium nitrogen in sea and estuarine waters. The efficiency of complexing agents to prevent precipitation of metallic hydroxides, due to the high pH value of the carrier solution, was studied. Under the optimised conditions, no interference was observed from different expected interfering ions as well as volatile amines. The proposed method provided the determination of NH4+ in concentrations ranging from 50 to 1000 microg L(-1), with detection and quantification limits of 18 and 35 microg L(-1), respectively. A determination rate of 20 h(-1) was achieved, with good repeatability for 10 consecutive injections of sea and estuarine samples (relative standard deviations lower than 2.0%). Accuracy of the methodology was assessed through recovery assays in 10 samples and also by analysis of certified reference material. PMID- 19137163 TI - Catalytic skeletal reorganization of enynes through electrophilic activation of alkynes: double cleavage of C-C double and triple bonds. AB - Skeletal reorganization of enynes was studied by electrophilic activation of alkynes with various metal catalysts; the products obtained can be classified into two types, one involving carbon-carbon bond single cleavage (type I) and the other involving carbon-carbon bond double cleavage (type II). Summarized in this review are our studies of the catalytic skeletal reorganization of enynes through the electrophilic activation of alkynes, and recent results from selective formation of type II products. PMID- 19137164 TI - Chemistry and structural determination of botcinolides, botcinins, and botcinic acids. AB - The first asymmetric total syntheses of botcinins C (18), D (19), E (20), and F (21), botcinic acid (22), botcinic acid methyl ester (23), botcineric acid (26), and 3-O-acetylbotcinic acid methyl ester () were achieved. The structures of these compounds have been unequivocally determined through their total syntheses and those of 20, 22, 23, 26, and are identified with the revised forms of the natural products formerly assumed to be 2-epibotcinolide (10), botcinolide (6), 4 O-methylbotcinolide (7), homobotcinolide (11), and 3-O-acetyl-5-O methylbotcinolide (8), respectively. It was further proved that the proposed nine membered ring structure of 2-epibotcinolide (10) is very unstable and the ineluctable translactonization easily occurred to form the corresponding gamma lactone . PMID- 19137165 TI - Switched role of fullerene in the Diels-Alder reaction: facile addition of dienophiles to the conjugated fullerene diene moiety. AB - The isolated cyclopentadiene moiety in a fullerene multi-adduct derivative reacts readily with dienophiles to form [4 + 2] cycloaddition products. PMID- 19137166 TI - Monoalkylation of primary amines and N-sulfinylamides. AB - An efficient monoalkylation of primary amines with primary or secondary alcohols catalyzed by Ra-Ni under mild conditions is described. PMID- 19137167 TI - 3-Mercaptopropionic acid-mediated synthesis of peptide and protein thioesters. AB - Peptides and proteins fragment sequence-specifically in the presence of 3 mercaptopropionic acid to afford thioesters which can be used in native chemical ligation reactions. PMID- 19137168 TI - Metal-organic replica of gamma-Pu: the first uninodal 10-connected coordination network based on pentanuclear cadmium clusters. AB - The first uninodal 10-connected metal-organic framework, based on pentanuclear cadmium cluster building blocks, exhibits an unprecedented gamma-Pu topology, which adds a new member to the series of metal-organic analogues which have a natural materials topology. PMID- 19137169 TI - Heteropolyacid-assisted fabrication of carbon nanostructures under ambient conditions. AB - Under ambient conditions, H(3)PW(12)O(40)-assisted electrochemical reduction process for chemical synthesis of SWNT-CNS, and H(3)PMo(12)O(40)-assisted ball milling process for COS fabrication. PMID- 19137170 TI - A new three-way supramolecular switch based on redox-controlled interconversion of hetero- and homo-guest-pair inclusion inside a host molecule. AB - A novel three-way supramolecular switch based on the interconversion of hetero guest-pair (D-A) and homo-guest-pair (D(2) or A(2)) inclusion inside cucurbit[8]uril is reported, which can be selectively controlled by chemical or electrochemical stimuli. PMID- 19137171 TI - Templated amplification of a naphthalenediimide-based receptor from a donor acceptor dynamic combinatorial library in water. AB - We report a dynamic combinatorial library that, upon binding of an electronically complementary guest, produces in high yield a tetrameric receptor with flat hydrophobic, electron-deficient surfaces and flexible, water-soluble disulfide containing linkers; analysis of the dependence of library composition on template concentration gives insight into the binding behaviours of the species involved. PMID- 19137172 TI - Unprecedented carbon-carbon bond cleavage in nucleophilic aziridine ring opening reaction, efficient ring transformation of aziridines to imidazolidin-4-ones. AB - N-Styryl-3-aryl-1-methylaziridine-2-carboxamides, which were readily obtained from the cross coupling reaction between 3-aryl-1-methylaziridine-2-carboxamides and 1-aryl-2-bromoethenes catalyzed by CuI/N,N-dimethylglycine in the presence of Cs(2)CO(3), underwent a base-mediated intramolecular nucleophilic aziridine ring opening reaction effectively via the carbon-carbon bond cleavage of aziridine to afford the ring expanded imidazolidin-4-one products in good yields. PMID- 19137173 TI - Dendronized macromonomers for three-dimensional data storage. AB - A series of dendritic macromonomers have been synthesized and utilized as the photoactive component in holographic storage systems leading to high performance, low shrinkage materials. PMID- 19137174 TI - beta-Diiminato complexes of arsenic including the formally AsI compound [As3L3] [L = [N(C6H3Pri2-2,6)C(H)]2CPh]. AB - The reaction of the potassium beta-diiminate KL (L = [[N(Ar)C(H)](2)CPh](-); Ar = C(6)H(3)Pr(i)(2)-2,6) with AsI(3) gave [AsI(2)L] (1), which upon reduction with KC(8) produced [(kappa(1)-L)(2)As-As[double bond, length as m-dash]As(kappa(2) L)] (2), having a unique bent chain of the three arsenic atoms in the formal oxidation states +2, 0 and +1. PMID- 19137175 TI - A lysine walk to high relaxivity collagen-targeted MRI contrast agents. AB - A strategy for preparing peptide-based magnetic resonance contrast agents with multiple gadolinium chelates is described. PMID- 19137176 TI - Synthesis of tri- and tetraynes using a butadiynyl synthon. AB - Using a one-pot protocol, triynes and tetraynes are formed from the reaction of a dibromovinyl triflate and a terminal alkyne under palladium-catalyzed cross coupling conditions. PMID- 19137177 TI - A cycloaddition route to novel triazole boronic esters. AB - The [3 + 2] cycloaddition of alkynylboronates and azides provides a direct route to novel triazole boronic esters, the regioselectivity of this process and functionalisation of the heterocycle products is described. PMID- 19137178 TI - Selective dimerization of 1,6-diynes catalyzed by ionic liquid-supported nickel complexes in an ionic liquid/toluene biphasic system. AB - A NiCl(2)-6H(2)O/Zn reagent with a 2-aryliminomethylpyridine ligand catalyzed cyclization/polymerization of 1,6-diynes to yield mainly the corresponding poly(1,3-butadienylene) compounds, whereas NiBr(2)-3H(2)O/Zn with the use of 3-(2 (2-methoxyethoxy)-ethyl)-1-methyl-1H-imidazol-3-ium bromide as a ligand converted 1,6-diynes to the corresponding annulated 1,3,5,7-cyclooctatetraenes in a biphasic solvent system consisting of toluene and an ionic liquid. PMID- 19137179 TI - Multiradiate calcium phosphate patterns derived from a gradating polysaccharide acidic protein system. AB - A series of multiradiate calcium phosphate patterns have been observed by a gradating chitosan-polyaspartatic acid (PAsp) system, and their morphology evolution reveals the effect of chitosan and PAsp on the interfacial biomineralization. PMID- 19137181 TI - Anthraquinone based polymer as high performance cathode material for rechargeable lithium batteries. AB - Poly(anthraquinonyl sulfide) is synthesized and investigated as a novel organic cathode material for rechargeable lithium batteries, which shows excellent reversibility and cyclabilty, and gives important insights into developing a new generation of organic cathode materials with higher performance. PMID- 19137180 TI - Tin triflate-mediated total synthesis of circumdatin F, sclerotigenin, asperlicin C, and other quinazolino[3,2-a][1,4]benzodiazepines. AB - Using tin triflate, as an effective Lewis acid, and microwaves, direct double cyclizations of bis(anthranilate)-containing tripeptide precursors to afford the total syntheses of 7-substituted quinazolino[3,2-a][1,4]benzodiazepinediones (1a f), including natural products circumdatin F (1a), sclerotigenin (1b), and asperlicin C (1c), were achieved with good overall isolated yields (23-62%). PMID- 19137182 TI - Highly regioselective ring-opening of trisubstituted aziridines by sulfur stabilised carbanions. AB - The highly regioselective, stereospecific ring-opening of trisubstituted N tosylaziridines possessing vinyl and hydroxymethyl groups by sulfone- and sulfide stablised carbanions is reported. PMID- 19137184 TI - Tunable coordination of a tropos phosphite for fine-tuning of the Rh catalyzed asymmetric addition of phenylboronic acid to cyclohexenone. AB - Using the deoxycholic acid derived tropos biphenylphosphite as a Rh(i) chiral ligand different complexes are obtained, depending on the Rh : L molar ratio, that give rise to the formation of different chiral products in the asymmetric addition of phenylboronic acid to cyclohexenone. PMID- 19137183 TI - Relaxation rates for spirocyclohexyl nitroxyl radicals are suitable for interspin distance measurements at temperatures up to about 125 K. AB - Between 65 and 175 K, nitroxyl radicals with spirocyclohexyl groups at the 2- and 6-positions of the piperidine ring exhibit spin echo dephasing rates that are slower than for nitroxyl radicals with 2,5-gem-dimethyl or 2,6-gem-dimethyl substituents that are currently used as spin labels, and are slow enough to permit DEER measurements at temperatures up to about 125 K for spin labels with analogous ring structures. PMID- 19137185 TI - Cyclisation of citronellal over heterogeneous inorganic fluorides--highly chemo- and diastereoselective catalysts for (+/-)-isopulegol. AB - Based on a fluorolytic sol-gel synthesis, nanoscopic metal fluorides and partly hydroxylated metal fluorides were synthesized; varying the F : OH ratio inside these solids yielded catalysts with different combinations and variable strength Lewis and Bronsted acid sites, which demonstrated unexpected catalytic properties for the diastereoselective synthesis of (+/-)-isopulegol. PMID- 19137186 TI - The asymmetric total synthesis of (-)-securinine. AB - The alkaloid (-)-securinine was synthesized in 18 steps and 16% overall yield from trans-4-hydroxy-l-proline. PMID- 19137187 TI - Diels-Alder cycloaddition as a new approach toward stable PC(sp3)P-metalated compounds. AB - A straightforward synthetic route toward a new family of dibenzobarrelene-based cyclometalated compounds is described. PMID- 19137188 TI - Me(2)Zn as a radical source in Reformatsky-type reactions. AB - Experimental evidence for the generation of radicals by Me(2)Zn used in Reformatsky reactions was unequivocally established with a radical trap. PMID- 19137189 TI - Seed cryopreservation of Halimium and Helianthemum species. AB - Seed germination of four Halimium species [H. atriplicifolium, H.hHalimifolium, H. ocymoides, H. umbellatum] and eight Helianthemum species [H. almeriense pau, H. appeninum.,H. cinereum, H. hirtum, H. marifolium, H. nummularium, H. syriacum, H. squamatum], all cistaceae species from the mediterranean region, was studied after seed storage in liquid nitrogen (Ln, -196C) for four months. In all samples assayed, mechanical scarification of the seed coat was carried out to enhance seed germination. For most samples studied, final germination percentages were unaffected by storage of seeds in Ln, both for intact seeds and for scarified seeds. The germination rate of cryopreserved seeds, expressed as days to reach 50 percent of the final germination percentage, was lower only for four samples of intact seeds and for three samples of scarified seeds. Therefore, this study shows that seed cryopreservation could be a suitable procedure for the long-term seed conservation of several Halimium and Helianthemum species. PMID- 19137190 TI - Comparative study of different mechanical models for identification of viscoelastic parameters of cryopreserved rabbit carotid arteries. AB - To characterize quantitatively the viscoelastic behaviors of cryopreserved rabbit carotid arteries, five mechanical models (a standard linear model, and four generalized Maxwell models) were comparatively applied. Specific aim of this study was to test the validity of these models to simulate the stress relaxation behaviors of the cryopreserved arteries and to find a best-fit model for identification of viscoelastic parameters. Non-linear curve fittings were applied to the stress-relaxation data measured with dynamic mechanical thermal analyzer (DMTA IV; Rheometric. Scientific Inc.; Piscataway, NJ, USA), it was found that 5 element model (generalized Maxwell body composed of an elastic element and two Kelvin body) could provide satisfactory description of the cryopreserved rabbit carotid arteries. PMID- 19137191 TI - Effects of glycerol pretreatment on recovery and antioxidant enzyme activities of lyophilized red blood cells. AB - Successful storage of red blood cells (RBCs) by freeze-drying technique has important implications in blood transfusion and clinical medicine. We presented a method of preservation RBCs by pretreating them with glycerol solution and then freeze-drying. The effects of glycerol pretreatment on recovery and antioxidant enzyme activities of lyophilized RBCs were investigated. Rehydration of pretreated in 40 percent glycerol and freeze-dried RBCs resulted in 55.3+/-4.3 percent numerical recovery and 53.5+/-3.9 percent hemoglobin recovery, which were significantly higher than freeze-dried RBCs without glycerol pretreatment (P less than 0.01). Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) activities of 40 percent glycerol pretreated and freeze-dried RBCs almost remained fully active after freeze-drying. After 180 days storage, the activities decreased by 25.6 percent , 21.5 percent and 18.4 percent, respectively, which are significantly less than those of freeze-dried RBCs without glycerol pretreatment (P < 0.1). These data demonstrated that glycerol pretreatment had beneficial effects on the recovery and antioxidant enzyme activities of lyophilized RBCs. PMID- 19137192 TI - Seasonal variations in antifreeze protein activity and haemolymph osmolality in larvae of the beetle Ragium mordax (Coleoptera: Cerambycidae). AB - Larvae of Rhagium mordax empty their guts in preparation for the winter, which alone may enable the larvae to supercool down to -20 degree C or below. This should be sufficient for the larvae to over winter in Denmark if they can prevent inoculation. Antifreeze proteins (AFP) prevent inoculation in adult Rhagium inquisitor and this is also likely in the larvae of R. mordax, as they are in contact with ice in their hibernacula during the winter. arvae of R. mordax probably produce AFPs in the early autumn, however, in some individuals thermal hysteresis (TH) as high as 5.01 degree C was observed in June. Whether or not these individuals have a constant level of TH in their haemolymph all year or if they produce further antifreeze proteins during the autumn is unknown. The lowest measured in January was 7.49 degree C (the highest during this month was 9.08 degree C) so it is likely that the individuals with the highest TH in June also produce AFPs. Haemolymph osmolality in R. mordax is relatively low compared to other freeze avoiding insects, samples taken in January peak at 741 mOsm (+/-127 mOsm). The results of this study are compared with similar data for the closely related R. inquisitor. PMID- 19137193 TI - Human ovarian tissue preservation: is vitrification acceptable method for assisted reproduction? AB - The aim of this study was to evaluate the most successful vitrification protocol. The ovarian tissue pieces were randomly distributed into seven groups including fresh control. Each experimental group was divided into three subgroups according to the following cooling modes: a) in 1.8 ml cryo-vials with 1ml vitrification medium, b) in 1.8 ml cryo-vials with 0.1 ml vitrification medium, or c) by direct dropping with 0.05 ml vitrification medium into liquid nitrogen. The best results were observed in the protocol using 2.62 M dimethylsulphoxide + 2.6 M acetamide + 1.31 M propylene glycol + 0.0075M polyethylene glycol in combination with direct dropping of ovarian tissue pieces into liquid nitrogen. The vitrified and rewarmed samples after in vitro culture with this protocol showed 86 percent normally developed follicles, compared with 92 percent in fresh non-treated control. The concentrations of hormones in spent medium from culture of the same samples were 319 pg/ml for 17beta-estradiol and 2.6 ng/ml for progesterone compared with fresh non-treated control (253 pg/ml and 6 ng/ml, respectively). The results obtained by vitrification of ovarian tissue with this protocol were compatible with those of the fresh ovarian tissue. PMID- 19137194 TI - Comparison of efficiency of open pulled straw (OPS) and Cryotop vitrification for cryopreservation of in vitro matured pig oocytes. AB - During the past few years vitrification has been acknowledged as a viable alternative to traditional slow-rate freezing in both animal and human embryology. However, few data are available regarding the comparative efficiency of published and commercially available vitrification methods. The purpose of our work was to compare the OPS and Cryotop technology for cryopreservation of porcine in vitro matured oocytes. In a 2 x 2 factorial experiment, OPS and Cryotop devices and solutions were used for vitrification and warming. Two hours after warming oocytes were parthenogenetically activated and cultured in vitro. In 6 replicates a total of 1153 oocytes were vitrified. The cleavage rate after vitrification with Cryotop device and Cryotop solution (34.7 percent) were higher than those after vitrification with Cryotop device and OPS solution, or OPS device with both OPS and Cryotop solution (11.5, 5.1 and 11.3 percent, respectively). Further embryo development has shown a similar difference: Cryotop device applied with Cryotop solution resulted in 11.6 percent blastocyst/oocyte rates, higher than those achieved with Cryotop device and OPS solutions, or OPS device with both Cryotop and OPS solution (1.6, 1,65 and 0.6 percent, respectively). Our results indicate that for cryopreservation of some highly sensitive biological specimen including porcine oocytes Cryotop vitrification is superior to the OPS technique. PMID- 19137195 TI - Cryopreservation of in vitro grown nodal segments of Rauvolfia serpentina by PVS2 vitrification. AB - This paper describes the cryopreservation by PVS2 vitrification of Rauvolfia serpentina (L.) Benth ex kurz, an important tropical medicinal plant. The effects of type and size of explants, sucrose preculture (duration and concentration) and vitrification treatment were tested. Preliminary experiments with PVS1, 2 and 3 produced shoot growth only for PVS2. When optimizing the PVS2 vitrification of nodal segments, those of 0.31 - 0.39 cm in size were better than other nodal sizes and or apices. Sucrose preculture had a positive role in survival and subsequent regrowth of the cryopreserved explants. Seven days on 0.5 M sucrose solution significantly improved the viability of nodal segments. PVS2 incubation for 45 minutes combined with a 7-day preculture gave the optimum result of 66 percent. Plantlets derived after cryopreservation resumed growth and regenerated normally. PMID- 19137196 TI - Cryopreservation of resting cysts of the freshwater ciliate Meseres corlissi by conventional two-step methods and one-step vitrification protocols. AB - Trophic cells of the freshwater ciliate Meseres corlissi CCAP 1647/1 proved to be recalcitrant to all the cryopreservation methods tested; however, resting cysts of this strain were amenable to both conventional two-step cryopreservation and ultra-rapid vitrification methods. Conventional controlled rate cooling and Mr. Frosty cooling methods, employing 5 percent dimethylsulphoxide (DMSO) as a cryoprotectant were effective in preserving cysts in either liquid medium or soil suspensions. Alternative cryopreservation methods involving dehydration of soil/cyst suspensions, in the absence of any colligitative cryoprotectant, and rapid cooling over liquid nitrogen or plunge freezing were effective. The level of residual moisture was the critical factor with no survival observed in any samples with > 35 percent residual moisture, and high levels of survival (> 50 percent) in samples with < 14 percent residual moisture. Trophic cells obtained from cryopreserved cysts appeared healthy and did not differ obviously from the controls with respect to morphology, movement, division and encystment/excystment reactions. In a test culture derived from material which had been cryopreserved by the rapid cooling method a maximum growth rate of 1.32 d(-1) (at 22 degree C) was determined, a value which agrees well with earlier observations on the original strain. PMID- 19137197 TI - Cryopreservation by encapsulation-dehydration of plumules of coconut (Cocos nucifera L.). AB - This study describes the use of an encapsulation-dehydration cryopreservation technique on coconut plumules (apical dome with three or four leaf primordia) excised from embryos. In order to establish a reliable cryopreservation process for plumules, several different key factors were tested: pretreatment duration, sugar concentration, dehydration period and freezing. In parallel, histological studies were performed to describe the structural changes of tissues and plumule cells subjected to dehydration and freezing. A good survival level of around 60% was obtained. However, after 8 months culture regrowth, this level decreased to a maximum of 20 % which was achieved using sucrose treatment. In this paper we report for the first time the regeneration of leafy shoots from coconut plumules after cryopreservation. PMID- 19137198 TI - Expression and antioxidant enzymes in Chaetoceros neogracile, an Antarctic alga. AB - We examined low temperature-induced protein profile alterations in the Antarctic alga Chaetoceros neogracile using a proteomic approach. Chaetoceros neogracile was cultured at 4 degree C and then cooled to 0 degree C, and the resultant cold induced alterations in protein expression patterns were analyzed by two dimensional gel electrophoresis. Of the approximately 150 protein spots detected by Coomassie staining, we identified 15 with a greater than two-fold change in amount. Of these, ten proteins were up-regulated and five were down-regulated after cold exposure. Three cellular protein quality control proteins, such as chaperone protein DnaK, chaperone ClpB, and 26S protease regulatory subunit 6B homolog were prominently increased, whereas chaperone protein HtpG was decreased in response to cold stress. Moreover, changes in enzyme activity and isozyme profiles for superoxide dismutase, glutathione reductase, and glutathione S transferase were also detected in the gel, using an enzyme activity staining method. These alterations in protein expression and antioxidant enzyme activity may be related to survival mechanisms of C. neogracile at low temperatures. PMID- 19137200 TI - A national strategy for the management of chronic obstructive pulmonary disease (COPD) in England: aiming to improve the quality of care for patients. AB - This article examines the rationale behind the development of the National Clinical Strategy for Chronic Obstructive Pulmonary Disease (COPD) in England which will be launched in 2009. It focuses on the burden which COPD places on the healthcare service and the effect it has on patients, and highlights the important steps in the development of the Clinical Strategy. The general recommendations which are likely to be contained within the Clinical Strategy are discussed, as well as the possible implications for primary care. PMID- 19137199 TI - Cognitive and behavioral status of low birth weight preterm children raised in a developing country at preschool age. AB - OBJECTIVE: To assess cognitive and behavioral development at preschool age of children born preterm and with low birth weight and raised in a developing country. METHODS: Prospective cross-sectional study of 80 neonates born in a university hospital in southern Brazil. Neuropsychological assessment at age 4-5 years included the Wechsler Preschool and Primary Scale of Intelligence (WPPSI), Conners' Parent Rating Scale-Revised, Denver test and neurological examination. Perinatal and early neuropsychomotor development information was collected from the database during follow-up. Results were related to birth weight, sex and gestational age. RESULTS: WPPSI scores were: total intelligence quotient (IQ) 88.00+/-16.96, verbal IQ 89.72+/-16.72, and executive IQ 88.12+/-15.71 for the group with less than 1,500 grams; and total IQ 91.11+/-14.73, verbal IQ 93.36+/ 12.65, and executive IQ 90.20+/-16.06 for the group between 1,500 and 2,500 grams. The best scores were obtained in tests that evaluated capacity of abstraction and symbolization, picture completion and common perception, in which only 5% and 6.3% of the children had abnormal results, respectively. The lowest scores were obtained in tests that evaluated visual-motor coordination and flexibility-speed of reasoning, in which 27.5% and 16.3% of the children had abnormal results, respectively. A total of 32.5% had abnormal results in the arithmetic tests. Behaviors related to attention deficit/hyperactivity disorder (ADHD) were observed in 48% of the sample. Low score in the Bayley Mental Scale and abnormal result in the Denver test were significantly correlated to ADHD (p = 0.017 and p = 0.004). Abnormal results in the Bayley Mental Scale (p < 0.001), Denver test (p < 0.001) and neurological examination (p = 0.002) were associated with lower IQ. CONCLUSIONS: The results revealed an increased incidence of behavioral and cognitive disorders at preschool age. PMID- 19137201 TI - Role of occupational hygiene research in the control of occupational health risks from engineered nanoparticles. PMID- 19137202 TI - Occupational loading, health behavior and sleep disturbance as predictors of low back pain. AB - OBJECTIVES: Risk factors for low-back pain are known to co-occur, but their joint effect has not often been studied. Little is also known about the variation of the risk factors or their effects with age. METHODS: This prospective study assessed the 1-year incidence of low-back pain by age group in a Finnish industrial population. The effects of the baseline variables on the risk of low back pain in the follow-up were estimated with a log-binomial regression. RESULTS: Among 2256 blue- and white-collar workers free of low-back pain 12 months preceding the baseline, 21% reported low-back pain after 1-year of follow up. Physical work load (sum of heavy lifting, awkward postures, and whole-body vibration) predicted low-back pain among those younger than 50 years [highest relative risk (RR) 2.4, 95% confidence interval (95% CI) 1.4-4.2], whereas health behavior (sum of smoking, overweight, and lack of physical exercise) increased the risk only among those 50 years or older (RR up to 2.8, 95% CI 1.4-5.4). Mental stress, dissatisfaction with life, and sleep problems were significant predictors in the group of 40- to 49-year-old workers. Work-related psychosocial factors were not associated with the outcome. CONCLUSIONS: In this study, workers of different ages were affected by slightly different risk factors. The results support the provision of health promotion and stress management as part of programs to prevent work-related low-back pain. In particular, aging workers may benefit from such an integrated approach. More prospective studies on the joint effects of age-specific risk factors of low-back pain are warranted. PMID- 19137203 TI - Predictors of sick leave and reduced productivity at work among persons with early inflammatory joint conditions. AB - OBJECTIVES: This study attempted to identify predictors of sick leave and reduced productivity at work among persons with early inflammatory joint conditions. METHODS: In a prospective cohort study of 210 workers with inflammatory joint conditions present for less than 12 months, data were collected by a medical examination and questionnaires at baseline and after 6 and 12 months. The outcomes were sick leave and reduced productivity at work. Generalized estimation equations (6-month time-lag model) were used to study predictors. RESULTS: Sick leave was predicted by high levels of pain [odds ratio (OR) 3.2], poor physical functioning (OR 4.4), and frequent manual materials handling (OR 2.0), whereas supervisors had a lower likelihood of sick leave (OR 0.2). The predictors of reduced productivity at work were intermediate levels of pain (OR 3.1), poor physical functioning (OR 2.8), poor mental health (OR 2.1), and low support from colleagues (OR 2.2), whereas the workers classified as having nonrheumatoid arthritis were less likely to report reduced productivity than those with inflammatory joint complaints without clinical synovitis (OR 0.4). CONCLUSIONS: Among the workers with early inflammatory joint conditions, self-reported pain and physical functioning affected performance at work, together with manual materials handling and lack of support from colleagues. Early treatment should target pain and physical functioning, and job interventions should aim at reducing physical workload and increasing coworker support. PMID- 19137204 TI - Influence of self-reported work conditions and health on full, partial and no return to work after long-term sickness absence. AB - OBJECTIVES: This study aimed at describing the frequency of full, partial, and no return to work after long-term sick leave and at ascertaining the influence of psychosocial work conditions, work ability and health, reported before the onset of sick leave, on full and partial return to work. METHODS: Altogether 853 public sector employees in Sweden, mainly women, with at least one sick leave lasting > or = 28 days, were studied. The outcome was the level of sick leave 2 years after the sick leave began. Potential predictors were self-rated health, work ability, and psychosocial work conditions assessed by questionnaire before the sick leave. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated by multinomial regression analyses. RESULTS: Altogether 41% of the participants went directly from full sick leave to full return to work; 21% had periods of partial return to work, but, at the 2-year follow-up, were fully back to work; 15% had partial return to work; and 23% were still not working. A relaxed work situation, a combination of low demands and high decision latitude, increased the odds for full (OR 2.72, 95% CI 1.60-4.62) and partial (OR 2.42, 95% CI 1.21-4.85) return to work. Negative consequences of organizational changes were associated with decreased odds for full return to work (OR 0.54, 95% CI 0.38-0.77). Good self rated health and work ability were associated with full return to work. CONCLUSIONS: Partial return to work often precedes full return to work, but also operates as a long-term solution for remaining occupationally active. Promoting relatively low demands and high decision latitude at work may support both full and partial return to work after long sick leaves. PMID- 19137205 TI - Sick leave among native and immigrant workers in Spain--a 6-month follow-up study. AB - OBJECTIVES: The incidence and the duration of sick leave were studied among immigrants and the native-born population in Spain. METHODS: This observational follow-up study included 1427 immigrants and 2793 Spanish natives treated at five primary care centers in Lleida in 2005 and followed for 6 months. The sick leave causes were coded according to the International Classification of Diseases (10th revision). Multivariate Poisson regressions estimated the rate ratio (RR) for sick leave adjusted for age, and linear regressions evaluated the effect of age, gender, and region of origin on the total number of sick-leave days. RESULTS: Altogether 19.5% of the natives and 12.7% of the immigrants had at least one sick leave episode. The incidence of new episodes per 100 person-years was lower for the immigrants than for the natives (32.5 versus 43.3 for the men and 18.6 versus 35.6 for the women, respectively). The mean duration of sick leave in the 6-month period was 19.4 (SD 29.4) days for the immigrants and 33.5 (SD 39.2) days for the natives. For the men, the risk of sick leave was greater for the natives than for the immigrants (adjusted RR 1.70, 95% confidence interval 1.43-2.02). After adjustment for age, the duration of sick leave for the native workers was 1.5 times greater than for the immigrants. CONCLUSIONS: Even though sick leave was less frequent among the immigrants than among the natives and the immigrant sick leave periods were of shorter duration, the two study populations did not show differences in the causes of disability. PMID- 19137206 TI - Exposure to iron and welding fumes and the risk of lung cancer. AB - OBJECTIVES: Exposure to iron fumes and dust and welding fumes is widespread and may increase the risk of lung cancer. The aim of this study was to identify associations between exposure to iron and welding fumes and the incidence of lung cancer among Finnish men. METHODS: The cohort of all economically active Finnish men, born in 1906-1945, who participated in the national census in 1970 was followed through the Finnish Cancer Registry for lung cancer cases (N=30,137) during 1971-1995. Their census occupations in 1970 were converted to estimates of cumulative exposure to iron and welding fumes with the Finnish job-exposure matrix on the basis of likelihood, average level, and estimated duration of exposure. Relative risk estimates for categorized cumulative exposure were defined by a Poisson regression, adjusted for smoking, socioeconomic status, and exposure to asbestos and silica dust. RESULTS: The relative risks for lung cancer increased as the cumulative exposure to iron and welding fumes increased. Relative risks in the highest exposure category was 1.35 [95% confidence interval (95% CI) 1.05-1.73] for iron and 1.15 (95% CI 0.90-1.46) for welding fumes. The respective relative risks estimated for squamous-cell carcinoma of the lungs were 1.94 (95% CI 1.35-2.78) and 1.55 (95% CI 1.08-2.24). There was no excess risk of small-cell carcinoma in any exposure category. CONCLUSIONS: Occupational exposure to iron and welding fumes was associated with an increase in lung cancer risk, mainly that of squamous-cell carcinoma. The simultaneous exposure to both of these agents and other potential work-related carcinogens complicates the interpretation of the independent roles of the risk factors. PMID- 19137207 TI - Behavior of aluminum in aluminum welders and manufacturers of aluminum sulfate- impact on biological monitoring. AB - OBJECTIVES: The suitability of determining aluminum in serum or urine as a form of biological monitoring was critically assessed. METHODS: Airborne and internal aluminum exposure was assessed for 12 aluminum welders in a shipyard and 5 manufacturers of aluminum sulfate. Particles were characterized with X-ray diffraction and scanning electron microscopy. Aluminum in air and biological samples was analyzed using electrothermal atomic absorption spectrometry. Basic toxicokinetic features were inferred from the data. RESULTS: The mean 8-hour time weighted average concentration of aluminum was 1.1 (range 0.008-6.1) mg/m(3) for the shipyard and 0.13 (range 0.02-0.5) mg/m(3) for the aluminum sulfate plant. Welding fume contained aluminum oxide particles <0.1 microm in diameter and their agglomerates, whereas bauxite and aluminum sulfate particles ranged from 1 to 10 microm in diameter. The shipyard welders' mean postshift serum and urinary concentrations of aluminum (S-Al and U-Al, respectively) were 0.22 and 3.4 micromol/l, respectively, and the aluminum sulfate workers' corresponding values were 0.13 and 0.58 micromol/l. Between two shifts, the welders' S-Al concentration decreased by about 50% (P<0.01), but their U-Al concentration did not change (P=0.64). No corresponding temporal changes occurred among the aluminum sulfate workers. After aluminum welding at the shipyard had ceased, the median S-Al concentration decreased by about 50% (P=0.007) within a year, but there was no change (P=0.75) in the corresponding U-Al concentration. CONCLUSIONS: About 1% of aluminum in welding fume appears to be rapidly absorbed from the lungs, whereas an undetermined fraction is retained and forms a lung burden. A higher fractional absorption of aluminum seems possible for aluminum sulfate workers without evidence of a lung burden. After rapid absorption, aluminum is slowly mobilized from the lung burden and dominates the S-Al and U-Al concentrations of aluminum welders. For kinetic reasons, S-Al or U-Al concentrations cannot be used to estimate the accumulation of aluminum in the target organs of toxicity. However, using U-Al analysis to monitor aluminum welders' lung burden seems practical. PMID- 19137208 TI - Long-term follow-up study of mortality and the incidence of cancer in a cohort of workers at a primary aluminum smelter in Sweden. AB - OBJECTIVES: Previous studies on mortality and the incidence of cancer among workers at primary aluminum smelters have produced conclusive results indicating an elevated risk of bladder cancer. An increased risk of lung cancer has also been reported several times. The objective of this study was to examine mortality and the incidence of cancer at a Swedish aluminum smelter when different neighboring reference populations were used to evaluate any relationships to the length of employment. METHODS: A historical cohort--comprised of 2264 male nonoffice workers employed from 1942 on and tracked up to the year 2000--was examined. With the use of three reference populations for mortality and four for cancer incidence, standardized mortality and incidence ratios were calculated, together with hazard ratios derived from Cox regression models. RESULTS: This study showed an excess risk of mortality due to chronic obstructive lung disease, mental disorders, and diseases of the digestive system among the short-term workers. An elevated risk of cancer was found for the lungs, central nervous system, and esophagus. The highest lung cancer risk was observed for the workers employed for > or = 10 years in the factory when they were compared with the reference group from northern Sweden (standardized incidence ratio 1.99, 95% confidence ratio 1.21-3.07). CONCLUSIONS: The results support previous studies that demonstrated an excess risk of lung cancer, but, in contrast to the results of most studies, cancer of the central nervous system was also elevated. This study did not, however, verify an association between this type of exposure and cancer of the urinary organs. PMID- 19137210 TI - Role of diagnoses and socioeconomic status in mortality among disability pensioners in Norway--a population-based cohort study. AB - OBJECTIVES: Several studies have shown increased mortality among disability pensioners. This study attempted to determine the causes of such an increase. METHODS: A population-based study was carried out with 148,942 persons followed between 1990 and 1996 in Norway. Of this total, 6285 women and 4113 men [corrected] were on a disability pension at baseline. A Cox proportional hazards analysis was carried out separately for the women and men in which all-cause mortality was the outcome variable. Disability pension status, disability pension diagnosis, age, educational level, and mean annual income were entered as explanatory variables. RESULTS: Persons on a disability pension had a strongly increased mortality rate. The age-adjusted hazard ratio was 3.0 [95% confidence interval (95% CI) 2.4-3.8] for the women and 3.4 (95% CI 2.8-4.1) for the men on a disability pension, when they were compared with those not on a disability pension. When adjusted for education and income levels, the hazard ratios (HR) decreased significantly for the men, to 2.0 (95% CI 1.8-2.4), but not so for women (HR 2.5, 95% CI 2.2-2.9). Except for the men with musculoskeletal diagnoses, all of the diagnostic groups had hazard ratios above unity also after the adjustments were made. CONCLUSIONS: The study confirmed high early mortality among Norwegian disability pensioners in the period 1990-1996. The medical condition seemed to contribute more to the increased mortality among the women, whereas a low socioeconomic status was more important for the men. PMID- 19137209 TI - Sharpening the focus on occupational safety and health in nanotechnology. AB - Increasing numbers of workers are involved with the production, use, distribution, and disposal of nanomaterials. At the same time, there is a growing number of reports of adverse biological effects of engineered nanoparticles in test systems. It is useful, at this juncture, to identify critical questions that will help address knowledge gaps concerning the potential occupational hazards of these materials. The questions address (i) hazard classification of engineered nanoparticles, (ii) exposure metrics, (iii) the actual exposures to the different engineered nanoparticles in the workplace, (iv) the limits of engineering controls and personal protective equipment with respect to engineered nanoparticles, (v) the kinds of surveillance programs that may be required at workplaces to protect potentially exposed workers, (vi) whether exposure registers should be established for workers potentially exposed to engineered nanoparticles, and, (vii) whether engineered nanoparticles should be treated as "new" substances and evaluated for safety and hazards? PMID- 19137211 TI - Does evening work predict sickness absence among female carers of the elderly? AB - OBJECTIVES: The aim of the present study was to predict the risk ratio of sickness absence lasting > or = 2 weeks due to shift work among Danish workers caring for the elderly during the evening and at night. METHODS: A sample of Danish carers of the elderly were interviewed in 2005. The response rate was 78%. A cohort of 5627 shift and day workers was followed for sickness absence lasting > or = 2 weeks and for sickness absence lasting > or = 8 weeks in a sickness compensation register covering all social transfer payments in Denmark. RESULTS: Among the evening workers, the rate ratio (RR) of sickness absence lasting > or = 2 weeks was 1.29 (95% confidence interval (95% CI) 1.10-1.52). The rate ratio for sickness absence lasting > or = 8 weeks was 1.24 (95% CI 0.99-1.56). CONCLUSIONS: Evening work may cause long-term sickness absence lasting > or = 2 weeks. PMID- 19137212 TI - [REACH--an ambitious project on the path to chemical safety]. PMID- 19137213 TI - [Historical development of the European legislation on chemicals]. AB - The development of the European legislation on chemicals must be viewed against the background of general developments in politics and society. Chemical accidents have played a triggering role, especially the catastrophe at Seveso in 1976. As a response, the "Seveso Directives" 82/501/EEC, 96/82/EEC, and 2003/105/EEC were passed. This also promoted general intentions to better comply with chemical hazards. The very first initiative of European institutions considering chemical compounds dates back to 1967 with Directive 67/548/EEC. Its amendments were later developed into the core of European legislation on chemicals. Other important fields of European legislation concerning chemicals, which were developed further, are environmental protection, consumer protection and occupational safety. PMID- 19137214 TI - [Risk assessment and chemical safety under REACH]. AB - Under the new European chemical legislation REACH (EG 1907/2006), dangerous properties of chemical substances will be evaluated and data gaps will be closed. This information is needed for other regulations, such as safety at the working place or for the safe handling of products. Existing knowledge on chemical compounds must be broadened because previous regulations have focused on high production volume compounds. The evaluation procedures needed too much time, and for the majority of non-evaluated substances a new strategy is needed. REACH places the duty for registration of substances with the producers and importers. Data gaps for risk evaluation will be closed. Competent authorities will be relieved from the primary risk evaluation of most substances. Alternative strategies of evaluation using previous information and non-animal testing approaches will be supported to avoid animal testing where appropriate. The new European chemical agency ECHA will use the provided information in order to identify very dangerous chemical substances which should be controlled by authorization of its use and the circumstances thereof. PMID- 19137215 TI - [Responsibilities of the European Chemical Agency under REACH]. AB - A new independent institution, the European Chemical Agency (ECHA), will be established after the new Regulation on Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) has come into effect, in order to ensure effective management at the community level. The Agency shall be managed by an executive director. The secretariat shall undertake all tasks related to the registration, the management of dossiers and the coordination of the evaluation of chemicals. The Agency also comprises a management board and committees for risk assessment, socio-economic analyses, a member state committee, a forum and a board of appeal. The Agency shall provide the Member States and the institutions of the Community with the best possible scientific and technical advice on questions related to chemicals. The innovations under REACH will introduce some flexibility, such as waiving of standard tests and simplification of administrative procedures, in order to gain resources. Until now no regulation on chemical safety has required such a large work load in such a short time with implemented new tasks and methods. PMID- 19137216 TI - [Legal aspects of the REACH regulation. The control system of the REACH regulation--new approaches in the EU chemical legislation]. AB - The REACH regulation from 2006 shall overcome the deficiencies of the previously existing inconsistent legal system of chemicals and build an efficient and innovative regulation for industrial chemicals in the EU. For this purpose, the REACH regulation is not inventing a completely new legislation for chemical substances, but refers to the existing rules, regulates and structures them in a new manner and complements them. With REACH a consistent control system for chemicals in Europe has been created, which basically is managed and coordinated by the newly established European Chemicals Agency (ECHA). In the first phases of the REACH system, information about chemicals is generated and afterwards evaluated. Then this information is used in a process of authorization and restriction, to ensure adequate proliferation and safe exposure to chemical substances. Numerous duties to furnish information complement the readjustment's procedural steps, particularly with regard to the supply chain and down to the consumer. It is mainly affected by the abrogation of the determination between new and existing substances, the principle of substitution and is based on the idea that industry itself is best suited to ensure that the substances it manufactures and places on the market in the EU do not adversely affect human health or the environment. PMID- 19137217 TI - [Evidence-based toxicological evaluation of chemicals by group summaries]. AB - REACH requires a certain amount of toxicological data to evaluate the registered chemicals. As a minimum, information on acute toxicity, skin and eye irritation, sensitisation, genotoxicity, organ specificity, the dose response and NOEL from repeated dose studies including toxicokinetics, reproductive toxicity and indication for carcinogenicity. In order to reduce the need for studies in animals, the use of several alternatives is proposed. In this communication the possible use of group summaries and the TTC (threshold of toxicological concern) concept are discussed. Although group summaries may bridge data gaps of structurally similar compounds by read across from better investigated chemicals, the examples presented indicate that for any group the read across needs to be thoroughly justified. The OECD documentation on group summaries provides some guidance. The TTC concept is another tool to reduce the need for animal testing. This may be applicable for compounds, for which human exposure is low and which are therefore not expected to be of relevance to human health. PMID- 19137218 TI - [The REACH legislation: the consumer and environment protection perspective]. AB - REACH has been initiated with the aim of improving existing legislation. In order to assist in the interpretation of the REACH legislation, guidance documents have been developed, which have only lately become available. According to the REACH annexes and supported by guidance documents, waiving of test requirements will be possible, thus, opening the possibility that under REACH no new (eco)toxicological data will be required. Concerning products, a guidance document was released in April 2008 stating that the substance concentration threshold of 0.1 % (w/w) applies to the article as produced or imported and it does not relate to the homogeneous materials or parts of an article, but relates to the article as such (i.e., as produced or imported). Hence, notification will not be required for many products containing chemicals with properties which place them on the candidate list for authorization. In summary, it is at present not foreseeable whether the expected benefit of the REACH legislation will materialise for the environment and for the health of consumers and at the work place. PMID- 19137219 TI - [Alternative methods to animal experiments. What can they afford in the safety testing of chemical substances under REACH?]. AB - Alternative methods to safety studies using laboratory animals have been accepted by the OECD in areas such as local toxicity and mutagenicity. In more complex important fields, such as systemic single and repeated dose toxicity, toxicokinetics, sensitisation, reproductive toxicity and carcinogenicity, it is expected that the development and validation of computerised methods, testing batteries (in vitro and in silico) and tiered testing systems will need many years and have to overcome many scientific and regulatory obstacles, which makes it extremely difficult to predict the outcome and the time needed. Therefore, the validated alternative methods available will only have a limited impact on reducing the numbers of animals required under REACH. In the midterm, the strategy should be more directed towards the refinement or reduction of in vivo testing because the replacement concerning complex toxicological endpoints is at present not in sight. PMID- 19137220 TI - [REACH: the guidance documents of the European Chemicals Agency (ECHA)]. AB - ECHA provides numerous Guidance Documents for the implementation of the REACH Regulation, emphasising that this Regulation is the only authentic legal reference. Furthermore, ECHA states that information in the Guidance Documents does not constitute legal advice and that public authorities do not accept any liability for the contents of these documents. Consequently, the companies themselves are responsible for the correct implementation of REACH. Because of their enormous volume and complexity, full knowledge of the contents of all Guidance Documents is not possible. Guidance Documents provide help with certain questions about REACH implementation. Companies complying with the REACH Regulation must be allowed to deviate from guidance. Furthermore, authorities cannot simply use the Guidance Documents for enforcement. Their decisions must be based on the Regulation. This publication describes the aims, goals and legal status of Guidance Documents. Problems concerning the comprehensibility of guidance and conformity with the REACH Regulation are discussed based on the examples of substance identification and chemical safety assessment. PMID- 19137221 TI - [Smoking cessation in Germany in 2007. Patterns of outpatient smoking cessation counselling and treatment]. AB - A qualified and comprehensive supply of outpatient smoking cessation counselling and treatment is essential to help smokers quit. In order to assess the status quo, structure and regional differences of the smoking cessation services available in Germany, a complete market survey was conducted in 2007. Descriptive results on the structure and characteristics of smoking cessation services show that there are distinct regional differences, especially in the New Laender, where the supply is insufficient. Overall, about a quarter of the providers of smoking cessation services lack specialised additional skills required for smoking cessation counselling, treatment or therapy of addicts. Especially medical practitioners more often lack such additional skills compared to other occupational groups. The smoking cessation services generally used the programs "Rauchfrei in 10 Schritten" and "Das Rauchfrei Programm", and the method of cognitive behavioural therapy. However, more than half of the services still use a method which is not evidence-based. Since February 2008 data on all supplies and suppliers of smoking cessation services, who gave their permission, have been published online and can be accessed at www.anbieter-raucherberatung.de. PMID- 19137223 TI - [Oxidative stress and the possibilities of measuring it for environmental medicine: Report of the Commission "Methods and Quality Assurance in Environmental Medicine"]]. PMID- 19137225 TI - [Regulations stability time of platelet concentration for the purpose of reducing life threatening septic transfusion reactions through bacterial contamination: from the 66th meeting of the Working Group on Blood on 9 June 2008 as adopted following (V 38)]. PMID- 19137226 TI - [Reduction of the risks associated with sepsis from the utilization of platelet transfusions: from the 66th meeting of the Working Group on Blood on 9 June 2008 as adopted following (V 38) ]. PMID- 19137227 TI - [10th notification of changes to the list of material preparation and disinfection processes pursuant to--11th drinking water regulation 2001 Standard: December 2008]]. PMID- 19137230 TI - The long-term outcome of laparoscopic colposuspension: a 10-year cohort study. AB - INTRODUCTION AND HYPOTHESIS: The aim of this study is to provide long-term outcome data, at least 10 years, following laparoscopic colposuspension. METHODS: The study includes a control group who underwent open colposuspension. A consecutive series of 139 women who had undergone laparoscopic colposuspension were reviewed and compared to 52 women who had an open colposuspension in the same unit. Subjects were contacted by telephone, at least 10 years post operatively, at which time a structured interview was performed which included the short-form Bristol Female Lower Urinary Tract Symptom questionnaire. RESULTS: There was deterioration in subjective cure rates from 71% and 67% at 6 months to 52% and 36% at 10 years for the laparoscopic and open procedures, respectively. CONCLUSION: This study provides evidence that laparoscopic colposuspension is probably as durable as open colposuspension. However, cure rates for both procedures appear to deteriorate over time, emphasising the importance of long term follow-up. PMID- 19137231 TI - Intravenous pamidronate in osteogenesis imperfecta type VII. AB - Cyclical intravenous treatment with pamidronate is widely used to treat osteogenesis imperfecta (OI) types I, III, and IV, which are due to dominant mutations affecting collagen type I alpha chains. There is no information about the effects of pamidronate in children with OI type VII, an autosomal-recessive form of OI caused by a mutation in the cartilage-associated protein gene. In this retrospective single-center study, we compared the effects of pamidronate in four girls with OI type VII (age range 3.9-12.7 years) to those in eight girls with OI types caused by collagen type I mutations who were matched for age and disease severity. During 3 years of pamidronate therapy, lumbar spine areal bone mineral density increased and lumbar vertebral bodies improved in shape in patients with OI type VII. Other outcomes such as fracture rates and mobility scores did not show statistically significant changes in this small study cohort. There were no significant side effects noted during the time of follow-up. Thus, intravenous treatment with pamidronate seems to be safe and of some benefit in patients with OI type VII. PMID- 19137232 TI - Imaging features of the total anomalous pulmonary venous connection to the portal vein shown by multidetector computed tomography angiography. PMID- 19137233 TI - Psychosurgery and deep brain stimulation as ultima ratio treatment for refractory depression. AB - For decades, the most severe, protracted and therapy-resistant forms of major depression have compelled clinicians and researchers to look for last resort treatment. Early psychosurgical procedures were hazardous and often associated with severe and persistent side effects including avolition, apathy and change of personality. With the introduction of psychopharmacological treatments in the 1950s, the frequency of ablative procedures declined rapidly. The past decade, however, has witnessed the resurgence of surgical strategies as a result of refined techniques and advances such as high frequency stimulation of deep brain nuclei. Recent data suggest that the overall effect of high frequency stimulation lies in the functional inhibition of neural activity in the region stimulated. Contrary to other psychosurgical procedures, high frequency stimulation reversibly modulates targeted brain areas and allows a postsurgical adaption of the stimulation parameters according to clinical outcome. With increased understanding of the brain regions and functional circuits involved in the pathogenesis of psychiatric disorders, major depression has emerged as a target for new psychosurgical approaches to selectively and precisely modulate neural areas involved in the disease process. Recent studies of minimally intervening procedures report good clinical outcome in the treatment of therapy-resistant forms of major depression. High frequency stimulation was successfully applied in several small samples of patients with treatment-resistant depression when the stimulation focused on different areas, e.g., nucleus accumbens, the lateral habenula or cortical areas. Nevertheless, the reticence toward psychosurgery, even for those patients suffering from the most debilitating forms of depression, still prevails, even though recent studies have shown significant improvement in terms of quality of life with the limitation that the number of treated cases has been small. In any event, valid and unambiguous criteria for patient eligibility have yet to be refined and standardized. In this review, we suggest possible standard criteria for the application of deep brain stimulation on patients suffering from otherwise treatment-resistant depression. PMID- 19137234 TI - Acquired stuttering after pallidal deep brain stimulation for dystonia. AB - We report two patients, in whom stuttering evolved as an adverse effect of pallidal deep brain stimulation for treating dystonia. Speech dysfluency was observed under conditions that optimally suppressed dystonic symptoms without inducing other extrinsic stimulation effects. This emphasizes a role of the sensorimotor part of the internal globus pallidus in regulating speech fluency. PMID- 19137235 TI - Serotonin transporter gene polymorphism (5-HTTLPR), environmental conditions, and developing negative emotionality and fear in early childhood. AB - Studies on neural and behavioral correlates of the serotonin transporter gene polymorphism (5-HTTLPR) strongly suggested interaction effects between the 5 HTTLPR genotype and environmental conditions on infant emotionality development. However, empirical studies that involve human infants are rare. The present study thus analyzed the interaction of the 5-HTTLPR genotype with the quality of maternal parenting behavior on the development of negative emotionality and fear in infancy. In a sample of 69 healthy firstborn infants, negative emotionality and fear were assessed at 4, 8, and 12 months using a multi-method approach. The quality of previous parenting has been operationalized as the quality of the mother-infant attachment relationship measured by the strange situation procedure at 18 months. Corresponding to hypotheses, to their caregiver insecurely attached infants who were homozygous for the s-variant of the 5-HTTLPR genotype developed a high level of negative emotionality and fear. The results thus are in line with the experimental results in the non-human primate model and point to a more pronounced susceptibility of s/s carrying infants to early rearing experiences. PMID- 19137236 TI - Sleep deprivation increases oleoylethanolamide in human cerebrospinal fluid. AB - This study investigated the role of two fatty acid ethanolamides, the endogenous cannabinoid anandamide and its structural analog oleoylethanolamide in sleep deprivation of human volunteers. Serum and cerebrospinal fluid (CSF) samples were obtained from 20 healthy volunteers before and after a night of sleep deprivation with an interval of about 12 months. We found increased levels of oleoylethanolamide in CSF (P = 0.011) but not in serum (P = 0.068) after 24 h of sleep deprivation. Oleoylethanolamide is an endogenous lipid messenger that is released after neural injury and activates peroxisome proliferator-activated receptor-alpha (PPAR-alpha) with nanomolar potency. Exogenous PPAR-alpha agonists, such as hypolipidemic fibrates and oleoylethanolamide, exert both neuroprotective and neurotrophic effects. Thus, our results suggest that oleoylethanolamide release may represent an endogenous neuroprotective signal during sleep deprivation. PMID- 19137237 TI - Anxiolytic effects of transcranial magnetic stimulation--an alternative treatment option in anxiety disorders? AB - In contrast to major depression, only few studies are available so far on the effects of repetitive transcranial magnetic stimulation (rTMS) in anxiety disorders. In order to summarise available data concerning the putative anxiolytic action of repetitive rTMS, a systematic literature review was carried out. Although interpretation of the results is difficult because of a large variety of used treatment protocols and the lack of a placebo-controlled design in the majority of studies, there is evidence for anxiolytic action of rTMS both from preclinical trials and studies in humans. Based on the idea of interhemispheric imbalance and/or deficits in cortico-limbic control as a model for human anxiety, inhibitory rTMS of the prefrontal cortex has been shown to exert beneficial effects in a number of studies in healthy subjects, patients with PTSD and panic disorder. However, to further elucidate the putative anxiolytic action of rTMS in patients with anxiety disorders future studies have to be conducted addressing in particular the limitations of the studies mentioned above. PMID- 19137239 TI - Recovery of ionizing-radiation damage after high doses of gamma ray in the hyperthermophilic archaeon Thermococcus gammatolerans. AB - The recently discovered hyperthermophilic and radioresistant archaeon Thermococcus gammatolerans is of great interest to compare and contrast the impact of its physiology on radioresistance and its ability to repair damaged chromosomes after exposure to gamma irradiation with radioresistant bacteria. We showed that, in contrast to other organisms, cell survival was not modified by the cellular growth phase under optimal growth conditions but nutrient-limited conditions did affect the T. gammatolerans radioresistance. We determined the first kinetics of damaged DNA recovery in an archaeon after exposure to massive doses of gamma irradiation and compared the efficiency of chromosomal DNA repair according to the cellular growth phase, nutrient availability and culture conditions. Chromosomal DNA repair kinetics showed that stationary phase cells reconstitute disrupted chromosomes more rapidly than exponential phase cells. Our data also revealed that this radioresistant archaeon was proficient to reconstitute shattered chromosomes either slowly or rapidly without any loss of viability. These results suggest that rapid DNA repair is not required for the extreme radioresistance of T. gammatolerans. PMID- 19137240 TI - Migraine strikes study: factors in patients' decision to treat early. AB - To describe factors associated with early treatment of migraine, and to examine reasons patients do not treat early, this cross-sectional observational study email-recruited migraineurs >or= 18-years-old who were currently prescribed acute migraine medication. Within 24 h of migraine resolution, eligible patients completed an online migraine strikes questionnaire which addressed pain severity, associated symptoms, and other variables including reasons for not treating early. Results reported were descriptive. Among 1,044 evaluable patients, early treatment was significantly associated with several factors such as leisure activity at onset (OR 1.32, P=0.010), photophobia (OR 1.39, P=0.013), diagnosis of migraine with aura (OR 1.36, P=0.004), and other factors. Among 840 patients who reported wanting to treat earlier desire to reserve medication for a severe migraine was the most common reason given for not doing so (51.2%). Overcoming these factors may facilitate earlier migraine treatment. PMID- 19137241 TI - Rapidly fatal necrotizing pneumonia in a 12-year-old boy caused by co-infection with parainfluenza virus type 1 and Panton-Valentine leukocidin (PVL)-positive methicillin-sensitive Staphylococcus aureus. PMID- 19137238 TI - Premenstrual mood symptoms: study of familiality and personality correlates in mood disorder pedigrees. AB - We sought to determine whether premenstrual mood symptoms exhibit familial aggregation in bipolar disorder or major depression pedigrees. Two thousand eight hundred seventy-six women were interviewed with the Diagnostic Interview for Genetic Studies as part of either the NIMH Genetics Initiative Bipolar Disorder Collaborative study or the Genetics of Early Onset Major Depression (GenRED) study and asked whether they had experienced severe mood symptoms premenstrually. In families with two or more female siblings with bipolar disorder (BP) or major depressive disorder (MDD), we examined the odds of having premenstrual mood symptoms given one or more siblings with these symptoms. For the GenRED MDD sample we also assessed the impact of personality as measured by the NEO-FFI. Premenstrual mood symptoms did not exhibit familial aggregation in families with BP or MDD. We unexpectedly found an association between high NEO openness scores and premenstrual mood symptoms, but neither this factor, nor NEO neuroticism influenced evidence for familial aggregation of symptoms. Limitations include the retrospective interview, the lack of data on premenstrual dysphoric disorder, and the inability to control for factors such as medication use. PMID- 19137242 TI - Erythromycin resistance in Streptococcus pyogenes and macrolide consumption in a central Italian region. AB - BACKGROUND: The study investigated macrolide resistance in Streptococcus pyogenes in a central Italian area from 2001 to 2006 and the possible correlation between antibiotic consumption and fluctuations of resistance percentages. MATERIALS AND METHODS: Macrolide and lincosamide susceptibility of 1,419 S. pyogenes isolates was tested by Kirby Bauer method. Macrolide consumption was evaluated as defined daily dose/1,000 inhabitants per day (DID), according to the World Health Organization anatomic therapeutic chemical classification. Spearman's correlation coefficient was used to assess the association between resistance and use of (1) all macrolides pooled, (2) once daily, (3) twice daily, and (4) three times daily dosage regimens. RESULTS: : In total, 320 strains (22.6%) were erythromycin resistant, 11.4% with the M phenotype and 11.2% with the MLS phenotype. There was a significant decrease in erythromycin resistance during the study period-from 28.1% in 2001 to 15.6% in 2006 (p < 0.01). No significant correlation was found between erythromycin resistance and local overall macrolide consumption, neither during the same year nor during the previous year. In contrast, a significant correlation was found between resistance rates and once-daily macrolide use during the preceding 6 months in Siena r = 0.747, p = 0.008). CONCLUSION: The known greater selective effect of long-acting agents could establish a pressure outcome, resulting in a specific local epidemiology during a relatively short time gap. PMID- 19137243 TI - [Atrial fibrillation -- a rhythmic problem with growing significance]. PMID- 19137244 TI - The German Competence Network on Atrial Fibrillation (AFNET). AB - The German Competence Network on Atrial Fibrillation (AFNET) is an interdisciplinary national research network funded by the Federal Ministry of Education and Research (BMBF) since 2003. The AFNET aims at improving treatment of atrial fibrillation (AF), the most frequent sustained arrhythmia of the heart. The AFNET has established a nationwide patient registry on manifestation, diagnostics, and therapy of AF in Germany. The data analyzed to date demonstrate that patients with AF are likely to have multiple comorbidities (hypertension, valvular heart disease, coronary artery disease, diabetes mellitus) and an advanced age. Regarding oral anticoagulation, guideline adherence is very high. Basic research has identified specific changes in atrial tissue during AF-induced remodeling providing the rationale for novel therapeutic interventions. Clinical trials are being carried out to optimize pharmacological and nonpharmacological treatments. The ANTIPAF trial is designed to prove that angiotensin II receptor blockers reduce the incidence of paroxysmal AF. The Flec-SL trial tests the efficacy of a short-term treatment with antiarrhythmic drugs after cardioversion. The Gap-AF trial investigates the impact of complete pulmonary vein (PV) isolation versus incomplete circumferential PV ablation on AF recurrences. The effect of preventive pacing on the recurrence of paroxysmal AF is studied in the BACE-PACE trial. PMID- 19137245 TI - [Anticoagulation in atrial fibrillation - an update]. AB - The correct anticoagulation regimen for prevention of thromboembolic events is essential in patients with atrial fibrillation. However, only a minority of patients receives anticoagulation according to the guidelines. The current guidelines are intended to make the indication for anticoagulation more simple and are summarized in the present article. This includes recommendations for chronic anticoagulation, prevention of thromboembolic events after cardioversion and in ablation of atrial fibrillation. PMID- 19137246 TI - New antiarrhythmic drugs for the treatment of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common arrhythmia requiring medical care, with a prevalence of almost 1% in the adult population. Particularly in the expanding elderly population, pharmacological therapy is and will continue to be the mainstay of AF therapy. Many currently used antiarrhythmic drugs have limited efficacy and cause cardiac and extracardiac toxicity. Thus, there is a continued need for development of new compounds with good efficacy and particularly with a favorable safety profile. Much emphasis is currently given to the development of so-called atrial-selective agents which target ion channels or proteins predominantly expressed in atrial myocardium. The rationale behind these compounds is to avoid unwanted effects on ionic currents on the ventricular site thus avoiding ventricular proarrhythmic effects. Alternatively, more conventional multichannel-blocking drugs are developed, for instance congeners of amiodarone. These molecules are designed to retain the electrophysiological efficacy of the mother compound while avoiding the extracardiac toxicity of this drug. The compounds which are far advanced in their clinical development are vernakalant ("atrial-selective") and dronedarone (multichannel-blocking). Preclinical and clinical findings of these substances are summarized. PMID- 19137248 TI - Balloon catheters for pulmonary vein isolation. AB - The mainstay of catheter ablation for atrial fibrillation (AF) is pulmonary vein isolation (PVI). The shortcomings of a point-by-point ablation approach using radiofrequency current steadily kindle the interest in new energy sources and catheter designs. The most promising currently available techniques are balloon catheters using cryothermal energy (CRYO) or high-intensity focused ultrasound (HIFU). Both technologies have proven to be efficacious. However, for both technologies treatment strategies have to be developed to overcome the relatively high incidence of collateral damage such as phrenic nerve palsy or atrial-to esophageal fistula. The results for patients with persistent AF in whom substrate modification is considered beneficial are poor and limit the use of balloon-based PVI to patients with paroxysmal atrial fibrillation (PAF). Moreover, based on the individual anatomy more than one balloon size may be required or may even make balloon-based PVI impossible in certain patients. PMID- 19137247 TI - [Rate versus rhythm control with and without heart failure]. AB - Atrial fibrillation and congestive heart failure are frequently associated with complex interactions. Patients with both diseases bear a sophisticated therapeutic challenge for the attending physician. The approach to treat atrial fibrillation differs for patients with and without heart failure in several aspects. Basic requirements are the treatment of the underlying diseases and prophylaxis of thromboembolic complications. Rate and rhythm control are the two main therapeutic strategies for atrial fibrillation according to the current guidelines. Large trials including the recently published AF-CHF study (Atrial Fibrillation - Congestive Heart Failure) failed to demonstrate a difference in mortality for both strategies. Thus, the therapeutic decision is mainly based on the patient's symptoms to improve quality of life. Rate control should be applied to asymptomatic patients or if rhythm control has already failed. If beta blockers and digoxin have failed to control heart rate, His ablation with pacemaker implantation can be considered. In patients without heart disease, class I antiarrhythmic drugs and, in case of ineffectiveness, amiodarone or catheter ablation are recommended for rhythm control. First data concerning catheter ablation of atrial fibrillation in heart failure are promising and randomized studies are on the way. Rhythm control remains first-line therapy in recent-onset or highly symptomatic paroxysmal or persistent atrial fibrillation patients with and without heart failure. PMID- 19137249 TI - Catheter ablation - new developments in robotics. AB - Catheter ablation has become the curative treatment modality for various arrhythmias. Extending the indications for catheter ablation from simple supraventricular tachycardias to complex arrhythmias such as ventricular tachycardia or atrial fibrillation, the investigator faces prolonged procedure times, fluoroscopy exposure and the need for stable and reproducible catheter movement. Recently, remote-controlled robotic catheter ablation has emerged as a novel ablation concept to meet these requirements. This review describes the two available robotic ablation systems and summarizes their clinical applications and current human experience. PMID- 19137250 TI - [Catheter interventions for congenital heart disease]. AB - Interventional catheterization for congenital heart disease has rapidly developed over the last 2 decades. The role of interventions has evolved from replacing surgery altogether in some cases to complementing surgery in others. The most commonly used approaches to common cardiac lesions such as persistent arterial duct, atrial and ventricular septal defects, valvar stenosis, coarctation of the aorta and pulmonary artery stenosis are discussed. PMID- 19137251 TI - [Severe acute left heart failure with pulmonary edema following cesarean section]. AB - BACKGROUND: Severe left heart failure shortly after delivery in a previously asymptomatic young woman can be caused by a peripartum cardiomyopathy, an exacerbation of valvular heart disease due to both congenital or acquired etiologies, a sustained chronic tachyarrhythmia (tachycardiomyopathy), a fulminant pulmonary embolism, or an infective endocarditis. CASE STUDY: 2 days following cesarean section, a 36-year-old primipara without known previous heart disease suffered from severe left heart failure with pulmonary edema (Figure 1) due to an acute bacterial aortic valve endocarditis. The infecting organisms were beta-hemolytic group A streptococci. After diagnosis could be confirmed by conventional Doppler echocardiography and by transesophageal echocardiography (Figures 2 and 3), the clinical situation of the patient stabilized in a few days under an initial antibiotic regimen with vancomycin and gentamicin, diuretics and catecholamines, followed by diuretics and ACE inhibitor. 14 days after cesarean delivery, aortic valve replacement could be performed under hemodynamically stable conditions. CONCLUSION: Although postpartal endocarditis is rarely described in the era of peripartum antibiotic prophylaxis, it should be considered in the differential diagnosis of patients with heart failure or fever in pregnancy, following delivery or cesarean section. The prognosis of peripartal endocarditis as a life-threatening disease is determined by an accurate and immediate diagnosis. PMID- 19137252 TI - [Angina pectoris and heart failure after coronary bypass operation. Documentation of an unusual cause]. PMID- 19137258 TI - Self and non-self. PMID- 19137259 TI - Transient and constitutive repression of cytoplasmic translation signaling in cells with mtDNA mutation. AB - Cytoplasmic translation is under sophisticated control but how cells adapt its rate to constitutive loss of mitochondrial oxidative phosphorylation is unknown. Here we show that translation is repressed in cells with the pathogenic A3243G mtDNA mutation or in mtDNA-less rho(0) cells by at least two distinct pathways, one transiently targeting elongation factor eEF-2 and the other initiation factor eIF-2alpha constitutively. Under conditions of exponential cell growth and mammalian target of rapamycin (mTOR) activation, eEF-2 becomes transiently phosphorylated by an AMP-activated protein kinase (AMPK)-dependent pathway, especially high in mutant cells. Independent of AMPK and mTOR, eIF-2alpha is constitutively phosphorylated in mutant cells, likely a signature of endoplasmic reticulum (ER)-stress response induced by the loss of oxidative phosphorylation. While the AMPK/eEF-2K/eEF-2 pathway appears to function in adaptation to physiological fluctuations in ATP levels in the mutant cells, the ER stress signified by constitutive protein synthesis inhibition through eIF-2alpha mediated repression of translation initiation may have pathobiochemical consequences. PMID- 19137260 TI - CCCTC-binding factor: to loop or to bridge. AB - Eukaryotic genomes have complex spatial organization in the nucleus. The factors and the mechanisms involved in this organization remain an enigma. Among the many proteins implicated in such a role, the ubiquitous Zn-finger protein CTCF stands out. Here we summarize the evidence placing CTCF in the enviable position of a master organizer of the genome. CTCF can form loops in cis, and can bridge sequences located on different chromosomes in trans. The thousands of CTCF binding sites, identified in recent genome-wide localization studies, and their distribution along the genome further support a crucial role of CTCF as a chromatin organizer. PMID- 19137261 TI - hShroom1 links a membrane bound protein to the actin cytoskeleton. AB - hShroom1 (hShrm1) is a member of the Apx/Shroom (Shrm) protein family and was identified from a yeast two-hybrid screen as a protein that interacts with the cytoplasmic domain of melanoma cell adhesion molecule (MCAM). The characteristic signature of the Shrm family is the presence of a unique domain, ASD2 (Apx/Shroom domain 2). mRNA analysis suggests that hShrm1 is expressed in brain, heart, skeletal muscle, colon, small intestine, kidney, placenta and lung tissue, as well a variety of melanoma and other cell lines. Co-immunoprecipitation and bioluminescence resonance energy transfer (BRET) experiments indicate that hShrm1 and MCAM interact in vivo and by immunofluorescence microscopy some co localization of these proteins is observed. hShrm1 partly co-localises with beta actin and is found in the Triton X-100 insoluble fraction of melanoma cell extracts. We propose that hShrm1 is involved in linking MCAM to the cytoskeleton. PMID- 19137262 TI - Galectin-3 stabilizes heterogeneous nuclear ribonucleoprotein Q to maintain proliferation of human colon cancer cells. AB - Comparative analysis of proteomes using 5-fluorouracil (5-FU)-resistant human colon cancer cell line revealed that decreased galectin-3 expression was significantly associated with retarded proliferation. However, in the presence of 5-FU proliferation rate of cells with suppressed galectin-3 expression did not differ from that of cells with normal galectin-3 expression, even galectin-3 suppression augmented apoptosis. Mechanism by which galectin-3 regulates cancer cell proliferation has been identified in immunoprecipitates of the anti-galectin 3 antibody. Heterogeneous nuclear ribonucleoprotein Q (hnRNP Q) was identified as a protein interacting with galectin-3. Interestingly, while galectin-3 protein was not affected by the hnRNP Q level, its suppression was accompanied by a decrease in hnRNP Q expression. The present study demonstrates that galectin-3 stabilizes hnRNP Q via complex formation, and reduction in the hnRNP Q level leads to slow proliferation and less susceptibility to 5-FU. PMID- 19137263 TI - Comparison of 90-s versus 5-min intraoperative 5-fluorouracil in trabeculectomy. AB - To compare the relative efficacy of trabeculectomy surgery with 90-s and 5-min intraoperative exposure time to 5-fluorouracil. This was a retrospective, non randomized comparative study. 41 eyes of 33 consecutive patients in the study group were compared to 40 eyes of 30 consecutive patients in the historical control group. Both groups were exposed to 5-fluorouracil (50 mg/ml) during trabeculectomy surgery. The exposure time was 90 s for the study group and 5 min for the control group. Three criteria were used to define surgical success: IOP (intraocular pressure) less than 21 mmHg; IOP less than 21 mmHg with more than 30% reduction in pressure; and IOP less than 15 mmHg with more than 30% reduction. The number of antiglaucoma medications, visual acuity, complications, and interventions were recorded at regular intervals. Mean preoperative IOP was 21.6 +/- 4.8 in the 90-s group and 21.2 +/- 4.9 in the 5-min group. Mean follow up was 28.2 +/- 5.1 months in the 90-s group and 48.0 +/- 4.9 months in the 5-min group. During the first 30 postoperative months, there were no statistically significant differences in IOP and requirement for antiglaucoma medications between the two groups. The 90-s group had shorter survival rates using the 1st success criteria, but no significant difference was detected when the more stringent 2nd and 3rd criteria were applied. Choroidal effusions were significantly less common in the 90-s group (P = 0.0076). The results of this small study suggest that a 90-s application of 5-fluorouracil may be as effective as a 5-min one in trabeculectomy. PMID- 19137264 TI - Automated protein structure calculation from NMR data. AB - Current software is almost at the stage to permit completely automatic structure determination of small proteins of <15 kDa, from NMR spectra to structure validation with minimal user interaction. This goal is welcome, as it makes structure calculation more objective and therefore more easily validated, without any loss in the quality of the structures generated. Moreover, it releases expert spectroscopists to carry out research that cannot be automated. It should not take much further effort to extend automation to ca 20 kDa. However, there are technological barriers to further automation, of which the biggest are identified as: routines for peak picking; adoption and sharing of a common framework for structure calculation, including the assembly of an automated and trusted package for structure validation; and sample preparation, particularly for larger proteins. These barriers should be the main target for development of methodology for protein structure determination, particularly by structural genomics consortia. PMID- 19137265 TI - Effect of acarbose on platelet-derived microparticles, soluble selectins, and adiponectin in diabetic patients. AB - Platelet-derived microparticles (PDMP), selectins, and adiponectin play an important role in the development of atherosclerosis in diabetes. Acarbose has been shown to have a beneficial effect on postprandial hyperglycemia in diabetic patients. However, its influence on PDMP, selectins, and adiponectin in these patients is poorly understood. We investigated the effect of acarbose on circulating levels of PDMP, selectins, and adiponectin in patients with type 2 diabetes. Acarbose (300 mg/day) was administered for 3 months. Levels of PDMP, sP selectin, sL-selectin, and adiponectin were measured by ELISA at baseline and after 1 and 3 months of treatment. The levels of PDMP, sP-selectin, and sL selectin were higher in diabetic patients than in hypertensive patients (PDMP; 35.1 +/- 34.2 vs. 53.3 +/- 56.7 U/ml, P < 0.05: sP-selectin; 134 +/- 52 vs. 235 +/- 70 ng/dl, P < 0.01: sL-selectin; 569 +/- 183 vs. 805 +/- 146 ng/ml, P < 0.05), while there were no significant differences between hypertensive and hyperlipidemic patients. Before acarbose treatment, the adiponectin level of diabetic patients was lower than that of hypertensive patients. Acarbose therapy significantly decreased the plasma PDMP level relative to baseline. Acarbose also caused a significant decrease of sP-selectin and sL-selectin. On the other hand, acarbose therapy led to a significant increase of adiponectin after 3 months of administration compared with baseline (adiponectin: diabetes versus hypertension, 3.61 +/- 1.23 vs. 5.87 +/- 1.92 microg/ml, P < 0.05; diabetes versus controls, 2.81 +/- 0.95 vs. 6.13 +/- 1.24 microg/ml, P < 0.01). Twelve of the 30 diabetic patients had a history of thrombotic complications. Furthermore, the reduction of PDMP and selectins during acarbose therapy was significantly greater in the thrombotic group (12 of 30) than in the nonthrombotic group (18 of 30) of diabetic patients. Acarbose may be beneficial for primary prevention of atherothrombosis in patients with type 2 diabetes. However, it requires a large clinical trial to test this hypothesis. PMID- 19137266 TI - Atrial natriuretic factor receptor guanylate cyclase signaling: new ATP-regulated transduction motif. AB - ANF-RGC membrane guanylate cyclase is the receptor for the hypotensive peptide hormones, atrial natriuretic factor (ANF) and type B natriuretic peptide (BNP). It is a single transmembrane spanning protein. Binding the hormone to the extracellular domain activates its intracellular catalytic domain. This results in accelerated production of cyclic GMP, a second messenger in controlling blood pressure, cardiac vasculature, and fluid secretion. ATP is the obligatory transducer of the ANF signal. It works through its ATP regulated module, ARM, which is juxtaposed to the C-terminal side of the transmembrane domain. Upon interaction, ATP induces a cascade of temporal and spatial changes in the ARM, which, finally, result in activation of the catalytic module. Although the exact nature and the details of these changes are not known, some of these have been stereographed in the simulated three-dimensional model of the ARM and validated biochemically. Through comprehensive techniques of steady state, time-resolved tryptophan fluorescence and Forster Resonance Energy Transfer (FRET), site directed and deletion-mutagenesis, and reconstitution, the present study validates and explains the mechanism of the model-based predicted transduction role of the ARM's structural motif, (669)WTAPELL(675). This motif is critical in the ATP-dependent ANF signaling. Molecular modeling shows that ATP binding exposes the (669)WTAPELL(675) motif, the exposure, in turn, facilitates its interaction and activation of the catalytic module. These principles of the model have been experimentally validated. This knowledge brings us a step closer to our understanding of the mechanism by which the ATP-dependent spatial changes within the ARM cause ANF signaling of ANF-RGC. PMID- 19137267 TI - Influence of methionine upon the concentration of malondialdehyde in the tissues and blood of rats exposed to sodium fluoride. AB - The aim of the study has been to determine the influence upon the kidney, liver, and the blood prooxidative system, exercised by administration of methionine (Met), under conditions of oxidative stress induced by sodium fluoride (NaF).The experiment was carried out on Wistar FL rats (adult females) that, for 35 days, were administered distilled water, NaF or NaF with methionine (doses: 10 mg NaF/kg bw/day, 10 mg Met/kg bw/day). The influence of administered NaF and Met was examined by analyzing the concentration of malondialdehyde (MDA) in kidney, liver, erythrocytes, and blood plasma.The study confirmed the disadvantageous effect of NaF upon the antioxidative system in rats (an increase in the concentration of MDA).The administration of methionine reduced the process of lipid peroxidation (a decreased in the concentration of MDA). The best antioxidative properties have been demonstrated by methionine in rat liver. PMID- 19137268 TI - Organized ribosome-containing structural domains in axons. AB - Periaxoplasmic ribosomal plaques (PARPs) are systematically recurring ribosome containing structural domains located in the F-actin-rich periphery of axoplasm in myelinated fibers. In contrast, endoaxoplasmic ribosomal plaques (EARPs) are small, oval-shaped ribosomal aggregate structures randomly dispersed within the axoplasm of unmyelinated squid giant axons. Ribosomes are attached to a superficial plaque-like structural matrix, which "caps" the domain at the outer cortical margin and appears fragmented in subcortical axoplasm. As such, the matrix represents a novel hallmark of PARP domains. Molecular markers concentrated in PARP domains include beta-actin mRNA, ZBP-1, SRP54, myosin Va and kinesin II molecular motor proteins. Rapid axoplasmic transport of microinjected heterologous radiolabeled BC1 RNA to putative PARP domains, mediated pari passu by microtubule- and F-actin-dependent systems, suggests that translation machinery, anchored by the matrix could provide targeted destinations for RNA trafficking. As distributed local centers of protein synthesis along axons, PARPs are likely to share modes of expression in common with other translational subdomains in neurons. PMID- 19137269 TI - The role of TNF in cancer. AB - Tumor necrosis factor (TNF) is an extraordinarily pleiotropic cytokine with a central role in immune homeostasis, inflammation, and host defense. Dependent on the cellular context, it can induce such diverse effects as apoptosis, necrosis, angiogenesis, immune cell activation, differentiation, and cell migration. These processes are of great relevance in tumor immune surveillance, and also play crucial roles in tumor development and tumor progression. It is therefore no surprise that TNF in a context-dependent manner displays pro- and antitumoral effects. Modulation of the activity of the TNF-TNF receptor system thus offers manifold possibilities for cancer therapy. In fact, TNF in combination with melphalan is already an established treatment option in the therapy of advanced soft tissue sarcoma of the extremities and many preclinical data suggest that TNF neutralization could also be exploited to fight cancer or cancer-associated complications. PMID- 19137270 TI - [Therapeutic aims for atrial fibrillation in patients with heart failure]. PMID- 19137272 TI - [Infections in hematology and oncology. Epidemiology--diagnosis--risk-adapted antimicrobial treatment--prevention]. AB - Infections are the predominant complications in patients with hematological and oncological diseases undergoing chemotherapy or stem cell transplantation. The spectrum of causative pathogens depends on the suppression of granulopoiesis, T cell immunity or humoral deficiency, so that recommendations for prevention, diagnostic approach and antimicrobial treatment should be specified for each cohort of patients. Following these rules, outcomes of prophylaxis and treatment today are favorable, although for some patient populations, the intensity of antimicrobial intervention has been reduced. Recommendations for prevention, including vaccination, should be known and followed for hospitalized as well as for out-patients. PMID- 19137273 TI - [Compliance with medication in epilepsy. Reply to the comments of Stefan H (2008) Nervenarzt 79:1446-1447]. PMID- 19137275 TI - [Innovative structuring in German care for schizophrenics]. AB - In a narrative literature review, innovative models of mental health care for people with schizophrenia are described. Structural aspects of mental health care such as the integration of services, new care paradigms, and innovative research questions are discussed. Key targets of innovative care include: improving continuity of care, introducing team-based community care, improving cooperation of all therapists, and integrating mental health, medical care, and social services. Models of current care for people with schizophrenia such as case management, crisis intervention, home treatment, and supported employment are described; and the evidence supporting these interventions is discussed. In the models of integrated care, there is an enhanced integration of psychosocial modules or service elements. One of the key challenges in mental health service systems is to consider the needs and aims of people with severe mental illness, to take into account biological vulnerability, symptoms of illness, and cognitive impairment of patients (whenever interventions are possible). One of the guiding principles is patient/user autonomy and a focus on user perspective. This implies that people suffering from schizophrenia should have the right to live in the least restrictive environment possible. PMID- 19137274 TI - [Symptoms and pathophysiological mechanisms of neuropathic pain syndromes]. AB - Neuropathic pain, caused by various central and peripheral nerve disorders, is especially problematic because of its severity, chronicity, and resistance to simple analgesics. Pathophysiologic changes responsible for generating pain following nerve lesions are usually independent of the etiology of the primary neuronal damage. Several fundamental mechanisms could be identified as principle conditions in the development of neuropathic pain; they include peripheral and central sensitization, disinhibition, sympathetically maintained pain, and cortical reorganization. Classifying neuropathic pain by these basic mechanisms is considered appropriate for basing new treatment approaches. In coming years, several advances are expected in the basic and clinical sciences of neuropathic pain which will provide new and improved therapies for patients continuing to experience this sometimes very disabling condition. PMID- 19137276 TI - [A 65-year-old female patient with gastric outlet obstruction of unknown origin]. AB - Gastric outlet obstructions can be of benign or malignant origin. In the case of the female patient described in this article, the extended diagnostic procedure with computed tomography and bone marrow biopsy was the key to a definite diagnosis and treatment planning. PMID- 19137277 TI - [Computed tomography of pancreatic tumors]. AB - Computed tomography (CT) and in particular multi-detector row computed tomography (MDCT), also known as multislice CT (MSCT), is ideally suited for detecting pancreatic tumors because of the high spatial resolution.The method of choice is hydro-CT which involves distension of the stomach and duodenum by administration of 1-1.5 l water as a negative contrast medium under medically induced hypotension by administration of buscopan. The patient is laid on the right side at an angle of 30-45 degrees in order to obtain an artefact-free image of the close anatomical relationship around the pancreas head. In addition, curved MPRs or in rare cases 3D reconstructions could be very helpful in identifying the critical anatomic tumor site in the neighbourhood of the visceral vessel system. After the correct diagnosis of an adenocarcinoma has been made only 20% of all patients are shown to have a surgically resectable disease, but the overall survival rate is significantly higher after resection in combination with a multimodal tumor therapy strategy. The reason is that the correct diagnosis of the resectability of the tumor is one of the main criteria for overall survival of these patients. Currently practically all pancreatic tumors can be detected using MDCT and the detection rate varies between 70% and 100% (most recent literature references give a sensitivity of 89% and specificity up to 99%). In some rare cases the differentiation between focal necrotizing pancreatitis and pancreatic carcinoma can be difficult even with sophisticated protocols. Resectability can be correctly diagnosed with MDCT with a sensitivity of 94% and a specificity of 89%. MDCT is an ideal tool for the detection of neuroendocrine tumors, metastases and for the differentiation of cystic pancreatic lesions such as pseudocysts, microcystic adenomas or intraductal papillary mucinous neoplasms (IPMN). Particularly, the differentiation of the latter into benign, borderline or malignant transformation is not always possible, but indirect signs, such as small nodules adjacent to the ductal wall, the diameter of the pancreatic duct, or a direct communication between cystic lesions and duct can be detected because of the high spatial resolution and is comparable to the findings in MRI. Moreover MD-CT is an ideal procedure for the differentiation of local tumor stages in patients under neoadjuvant or adjuvant chemotherapy. PMID- 19137278 TI - Outcome of double-bundle ACL reconstruction using hamstring tendons. AB - The objective of this article is to describe the clinical outcome of the transitional double-bundle procedure for anatomical ACL reconstruction. Subjects included 78 patients (average age 25 years) who had undergone ACL reconstruction with the transitional double-bundle procedure with multi-stranded hamstring tendons. The femoral socket for the anteromedial (AM) graft was created at 5-6 mm from the mid-sagittal line of the intercondylar notch at 2:00 or 10:00, and that for the posterolateral (PL) graft was drilled adjacent to the AM socket at 3:00 or 9:00. For the tibial side, two tunnels were made at the center of the footprint of the AM and PL bundles of the normal ACL. Patients were evaluated at 24 months or longer postoperatively based on the IKDC Knee Examination Form. Subjectively, 32 knees (41%) were graded as normal; 41 (53%), as nearly normal; 4 (5%), as abnormal; and 1 (1%) as graft rupture by re-injury. The average side-to side difference in anterior laxity at manual maximum force with the KT-2000 arthrometer was 0.9 mm +/- 1.2. Seventy patients (93%) had a range between -1 mm and 2 mm. In conclusion the transitional double-bundle ACL reconstruction provided a satisfactory outcome after a short-term follow-up. PMID- 19137279 TI - Positron emission tomographic measure of brain dopamine dependence to nicotine as a model of drugs of abuse. AB - RATIONALE: Nicotine/tobacco are prototypic substances used throughout the world. Nicotine abstinence produces some depressive-like effects which are treated by the dopamine (DA) and norepinephrine reuptake inhibitor bupropion. A quantitative measure of the regional brain utilization of these catecholamines (CA) during nicotine dependence and withdrawal is important. OBJECTIVE: The aim of this study was to prove that regional brain DA utilization by nicotine can be quantified by positron emission tomography (PET) using L-[beta-(11)C]DOPA. MATERIALS AND METHODS: Eight young Macaca mulatta monkeys were given 0.9% NaCl or nicotine in doses of 32 or 100 microg/kg i.m. bid for 9 days to produce minimal dependence. On the tenth day, PET measurements were repeated before and after i.v. nicotine administration. PET studies were done in habituated, trained, and fully conscious animals. RESULTS: Compared to a 0.9% NaCl control, acute i.v. nicotine as a bolus plus infusion for 30 min in similar doses to maintain a steady-state level for 30 min did not affect the utilization rate constant (k (3)) in dorsal or ventral striatum as measured by L-[beta-(11)C]DOPA. When monkeys were given nicotine bid repeatedly after overnight nicotine abstinence, CA utilization was reduced. A subsequent nicotine dose normalized utilization to slightly above control levels. Changes in ventral striatum were similar to those in dorsal striatum. The reduced rate of utilization demonstrated with L-[beta-(11)C]DOPA after overnight nicotine abstinence and its reversal by nicotine the next day provides an important PET measure of brain nicotine dependence and withdrawal. This method can be applied to other substances of abuse that release DA. PMID- 19137280 TI - Methylphenidate improves response inhibition but not reflection-impulsivity in children with attention deficit hyperactivity disorder (ADHD). PMID- 19137281 TI - Problem-oriented drug information: physicians' expectations and impact on clinical practice. AB - PURPOSE: Problem-oriented drug information (POD) is a service in which health professionals provide evidence-based answers to clinical questions posed by physicians. The objective of this study was to evaluate the user satisfaction and clinical impact of POD, to investigate predictors for use and to examine the kind of sources physicians search before applying for POD. METHODS: To evaluate POD, a questionnaire was distributed with problem-oriented answers sent from a drug information centre to physicians during the period of April 2006 to March 2007. RESULTS: Of 197 questionnaires, 183 (93%) were returned. The information from the POD service was highly valued by the physicians, and 90% of the answers led to reported impact on clinical practice in the specific clinical situation. Furthermore, 74% of the answers were intended to be used in a wider context either for future patients (67%) or for dissemination to colleagues (51%). Secondary-care physicians more often than general practitioners (GPs) used the information for dissemination to colleagues (63 vs. 39%, P = 0.0008), while GPs more often used the answer to support patient information (88 vs. 70%, P = 0.0029). The most prominent motive for applying for POD was a request for evidence-based information (78%), and the service was used to overcome barriers to practicing evidence-based medicine such as lack of time (36%), skills for searching (26%), and appraising the literature (13%). Before inquiring, 74% of the physicians had tried other information sources; the most frequent sources used were a drug reference (68%) and consulting a colleague (24%). Secondary-care physicians reported fewer barriers than GPs when seeking information, and secondary-care physicians searched other sources more often than GPs before contacting the service (81 vs. 67%, P = 0.031). CONCLUSION: POD represents a useful source for acquiring evidence-based drug information by physicians. POD is highly valued by the users. It was reported to have an impact on clinical practice for the specific patient but is also intended to be used in a wider context for future patients or for dissemination to colleagues. GPs' and secondary-care physicians' use of POD differs with GPs having more focus on patient information and secondary-care physicians having more focus on dissemination of the information to colleagues. PMID- 19137282 TI - Diagnostic accuracy and additional value of diffusion-weighted imaging for discrimination of malignant cervical lymph nodes in head and neck squamous cell carcinoma. AB - INTRODUCTION: The aim was to determine the diagnostic accuracy and additional value of diffusion-weighted imaging for detection of malignant lymph nodes in head and neck squamous cell carcinoma. METHODS: Two hundred nineteen lymph nodes, predominantly smaller than 10 mm (95.4%), in 16 consecutive patients were evaluated at 1.5 T. Lymph nodes were evaluated for maximum short axial diameter, morphological criteria, and apparent diffusion coefficient (ADC) values (b = 0 and b = 1,000 s/mm(2)). Sensitivity, specificity, positive and negative predictive values as well as diagnostic odds ratios (DORs) and areas under the curves (AUCs) of ROC curves were calculated for the various magnetic resonance imaging (MRI) criteria individually and in combination. Histological examination of lymph nodes in the neck dissection specimen was the gold standard to determine malignant involvement. RESULTS: The optimal ADC threshold was 1.0 x 10(-3) mm(2)/s. Using this cutoff point, sensitivity and specificity were 92.3% and 83.9%, respectively. When used in combination with size and morphological criteria, ADC value <1.0 x 10(-3) mm(2)/s was the strongest predictor of presence of metastasis (DOR = 97.6). A model which added ADC values to the other MRI criteria performed significantly better than a model without ADC values: AUC = 0.98 versus AUC = 0.91 (p = 0.036). CONCLUSION: In this study, with predominantly small lymph nodes, the ADC criterion is the strongest independent predictor of presence of metastasis. The use of ADC values in combination with the other MRI criteria significantly improves the discrimination between malignant and benign lymph nodes. PMID- 19137283 TI - Radiological findings of isolated hepatic tuberculosis in a child with tubulointerstitial nephritis-uveitis syndrome. AB - Isolated hepatic tuberculosis is a rare manifestation of tuberculosis in children although its incidence has been increasing, especially in immunosuppressed patients. Imaging studies frequently present a diagnostic challenge, especially for the nodular form. Thus a high index of suspicion is required for diagnosis. On US, hepatic tuberculosis may rarely present as a macronodular form (also called pseudotumour or tuberculoma). We report a 15-year-old girl with multiple hepatic macronodular masses due to isolated hepatic tuberculosis. PMID- 19137284 TI - Photogeneration of membrane potential hyperpolarization and depolarization in non excitable cells. AB - We monitored femtosecond laser induced membrane potential changes in non excitable cells using patchclamp analysis. Membrane potential hyperpolarization of HeLa cells was evoked by 780 nm, 80 fs laser pulses focused in the cellular cytoplasm at average powers of 30-60 mW. Simultaneous detection of intracellular Ca2+ concentration and membrane potential revealed coincident photogeneration of Ca2+ waves and membrane potential hyperpolarization. By using non-excitable cells, the cell dynamics are slow enough that we can calculate the membrane potential using the steady-state approximation for ion gradients and permeabilities, as formulated in the GHK equations. The calculations predict hyperpolarization that matches the experimental measurements and indicates that the cellular response to laser irradiation is biological, and occurs via laser triggered Ca2+ which acts on Ca2+ activated K+ channels, causing hyperpolarization. Furthermore, by irradiating the cellular plasma membrane, we observed membrane potential depolarization in combination with a drop in membrane resistance that was consistent with a transient laser-induced membrane perforation. These results entail the first quantitative analysis of location dependent laser-induced membrane potential modification and will help to clarify cellular biological responses under exposure to high intensity ultrashort laser pulses. PMID- 19137285 TI - Effects of NO2(-) and NO3(-) on the Fe(III)EDTA reduction in a chemical absorption-biological reduction integrated NO(x) removal system. AB - The biological reduction of Fe(III) ethylenediaminetetraacetic acid (EDTA) is a key step for NO removal in a chemical absorption-biological reduction integrated process. Since typical flue gas contain oxygen, NO(2)(-) and NO(3)(-) would be present in the absorption solution after NO absorption. In this paper, the interaction of NO(2)(-), NO(3)(-), and Fe(III)EDTA reduction was investigated. The experimental results indicate that the Fe(III)EDTA reduction rate decrease with the increase of NO(2)(-) or NO(3)(-) addition. In the presence of 10 mM NO(2)(-) or NO(3)(-), the average reduction rate of Fe(III)EDTA during the first 6-h reaction was 0.076 and 0.17 mM h(-1), respectively, compared with 1.07 mM h( 1) in the absence of NO(2)(-) and NO(3)(-). Fe(III)EDTA and either NO(2)(-) or NO(3)(-) reduction occurred simultaneously. Interestingly, the reduction rate of NO(2)(-) or NO(3)(-) was enhanced in presence of Fe(III)EDTA. The inhibition patterns observed during the effect of NO(2)(-) and NO(3)(-) on the Fe(III)EDTA reduction experiments suggest that Escherichia coli can utilize NO(2)(-), NO(3)( ), and Fe(III)EDTA as terminal electron acceptors. PMID- 19137286 TI - Efficient generation of recessive traits in diploid sake yeast by targeted gene disruption and loss of heterozygosity. AB - Sake yeast, a diploid Saccharomyces cerevisiae strain, is useful for industry but difficult to genetically engineer because it hardly sporulates. Until now, only a few recessive mutants of sake yeast have been obtained. To solve this problem, we developed the high-efficiency loss of heterozygosity (HELOH) method, which applies a two-step gene disruption. First, a heterozygous disruptant was constructed by gene replacement with URA3, followed by marker recycling on medium containing 5-fluoroorotic acid (5-FOA). Subsequently, spontaneous loss of heterozygosity (LOH) yielding a homozygous disruptant was selected for in a second round of gene integration. During this step, the wild-type allele of the heterozygous disruptant was marked by URA3 integration, and the resulting transformants were cultivated in non-selective medium to induce recombination and then grown on medium with 5-FOA to enrich for mutants that had undergone LOH. Although the frequency with which LOH occurs is extremely low, many homozygous disruptants were obtained with the HELOH method. Thus, we were able to efficiently construct homozygous disruptants of diploid sake yeast without sporulation, and sake yeast strains with multiple auxotrophies and a protease deficiency could be constructed. The HELOH method, therefore, facilitated the utilization of diploid sake yeast for genetic engineering purposes. PMID- 19137287 TI - Indigenous microfungi and plants reduce soil nonylphenol contamination and stimulate resident microfungal communities. AB - Nonylphenol, the most abundant environmental pollutant with endocrine disrupting activity, is also toxic to plants and microorganisms, but its actual impact in the field is unknown. In this study, diversity of culturable soil microfungal and plant communities was assessed in a disused industrial estate, at three sites featuring different nonylphenol pollution. Although soil microfungal assemblages varied widely among the sites, no significant correlation was found with point pollutant concentrations, thus suggesting indirect effects of soil contamination on microfungal assemblages. The potential of indigenous fungi and plants to remove nonylphenol was assessed in mesocosm experiments. Poplar plants and a fungal consortium consisting of the most abundant strains in the nonylphenol polluted soil samples were tested alone or in combination for their ability to reduce, under greenhouse conditions, nonylphenol levels either in a sterile, artificially contaminated sand substrate, or in two non-sterile soils from the original industrial area. Introduction of indigenous fungi consistently reduced nonylphenol levels in all substrates, up to ca. 70% depletion, whereas introduction of the plant proved to be effective only with high initial pollutant levels. In native non-sterile soil, nonylphenol depletion following fungal inoculation correlated with biostimulation of indigenous fungi, suggesting positive interactions between introduced and resident fungi. PMID- 19137288 TI - Optimization of actinomycin V production by Streptomyces triostinicus using artificial neural network and genetic algorithm. AB - Artificial neural network (ANN) and genetic algorithm (GA) were applied to optimize the medium components for the production of actinomycin V from a newly isolated strain of Streptomyces triostinicus which is not reported to produce this class of antibiotics. Experiments were conducted using the central composite design (CCD), and the data generated was used to build an artificial neural network model. The concentrations of five medium components (MgSO(4), NaCl, glucose, soybean meal and CaCO(3)) served as inputs to the neural network model, and the antibiotic yield served as outputs of the model. Using the genetic algorithm, the input space of the neural network model was optimized to find out the optimum values for maximum antibiotic yield. Maximum antibiotic yield of 452.0 mg l(-1) was obtained at the GA-optimized concentrations of medium components (MgSO(4) 3.657; NaCl 1.9012; glucose 8.836; soybean meal 20.1976 and CaCO(3) 13.0842 gl(-1)). The antibiotic yield obtained by the ANN/GA was 36.7% higher than the yield obtained with the response surface methodology (RSM). PMID- 19137290 TI - Continuous 2-keto-L-gulonic acid fermentation from L-sorbose by Ketogulonigenium vulgare DSM 4025. AB - A single-stage continuous fermentation process for the production of 2-keto-L gulonic acid (2KGA) from L-sorbose using Ketogulonigenium vulgare DSM 4025 was developed. The chemostat culture with the dilution rate that was calculated based on the relationship between the 2KGA production rate and the 2KGA concentration was feasible for production with high concentration of 2KGA. In this system, 112.2 g/L of 2KGA on the average was continuously produced from 114 g/L of L sorbose. A steady state of the fermentation was maintained for the duration of more than 110 h. The dilution rate was kept in the range of 0.035 and 0.043 h( 1), and the 2KGA productivity was 3.90 to 4.80 g/L/h. The average molar conversion yield of 2KGA from L-sorbose was 91.3%. Under the optimal conditions, L-sorbose concentration was kept at 0 g/L. Meanwhile, the dissolved oxygen level was changing in response to the dilution rate and 2KGA concentration. In the dissolved oxygen (DO) range of 16% to 58%, it was revealed that the relationship between DO and D possessed high degree of positive correlation under the L sorbose limiting condition (complete consumption of L-sorbose). Increasing D closer to the critical value for washing out point of the continuous fermentation, DO value tended to be gradually increased up to 58%. In conclusion, an efficient and reproducible continuous fermentation process for 2KGA production by K. vulgare DSM 4025 could be developed using a medium containing baker's yeast without using a second helper microorganism. PMID- 19137289 TI - Addition of an N-terminal epitope tag significantly increases the activity of plant fatty acid desaturases expressed in yeast cells. AB - Saccharomyces cerevisiae shows great potential for development of bioreactor systems geared toward the production of high-value lipids such as polyunsaturated omega-3 fatty acids, the yields of which are largely dependent on the activity of ectopically expressed enzymes. Here, we show that the addition of an N-terminal epitope tag sequence (either Myc or hemagglutinin) to oleate desaturase (FAD2) or omega-3 linoleate desaturase (FAD3) enzymes from plants, which catalyze consecutive reactions in the production of long chain omega-3 fatty acids, significantly increases their activity up to fourfold when expressed in yeast cells. Quantitative protein blotting using an antibody specific for native FAD2 revealed that the steady-state amount of the epitope-tagged FAD2 protein was also approximately fourfold higher than that of its untagged counterpart, demonstrating a direct relationship between the epitope tag-induced increase in enzyme amount and fatty acid product formation. Protein half-life and RNA blotting experiments indicated that the half-lives and mRNA content of the tagged and untagged FAD2 proteins were essentially the same, suggesting that the epitope tags increased protein abundance by improving translational efficiency. Taken together, these results indicate that the addition of an epitope tag sequence to a plant fatty acid desaturase (FAD) not only provides a useful means for protein immunodetection using highly specific, commercially available antibodies, but that it also significantly increases FAD activity and the production of polyunsaturated fatty acids in yeast cells. PMID- 19137291 TI - Diagnosing pulmonary embolism: clinical problem or methodological issue? PMID- 19137292 TI - Molecular imaging of alpha7 nicotinic acetylcholine receptors: design and evaluation of the potent radioligand [18F]NS10743. AB - PURPOSE: The outstanding diversity of cellular properties mediated by neuronal and nonneuronal alpha7 nicotinic acetylcholine receptors (alpha7 nAChR) points to the diagnostic potential of quantitative nuclear molecular imaging of alpha7 nAChR in neurology and oncology. It was our goal to radiolabel the alpha7 nAChR agonist 4-[5-(4-fluoro-phenyl)-[1,3,4]oxadiazol-2-yl]-1,4-diaza bicyclo[3.2.2]nonane (NS10743) and to assess the selectivity of [(18)F]NS10743 binding site occupancy in animal experiments. METHODS: [(18)F]NS10743 was synthesized by nucleophilic substitution of the nitro precursor. In vitro receptor affinity and selectivity were assessed by radioligand competition and autoradiography. The radiotracer properties were evaluated in female CD-1 mice by brain autoradiography and organ distribution. Target specificity was validated after treatment with SSR180711 (10 mg/kg, intraperitoneal), and metabolic stability was investigated using radio-HPLC. RESULTS: The specific activity of [(18)F]NS10743 exceeded 150 GBq/micromol at a radiochemical purity >99%. In vitro, NS10743 and [(18)F]NS10743 showed high affinity and specificity towards alpha7 nAChR. The brain permeation of [(18)F]NS10743 was fast and sufficient with values of 4.83 and 1.60% injected dose per gram and brain to plasma ratios of 3.83 and 2.05 at 5 and 60 min after radiotracer administration. Brain autoradiography and organ distribution showed target-specific accumulation of [(18)F]NS10743 in brain substructures and various alpha7 nAChR-expressing organs. The radiotracer showed a high metabolic stability in vivo with a single polar radiometabolite, which did not cross the blood-brain barrier. CONCLUSION: The good in vitro and in vivo features of [(18)F]NS10743 make this radioligand a promising candidate for quantitative in vivo imaging of alpha7 nAChR expression and encourage further investigations. PMID- 19137293 TI - Intraindividual comparison of 68Ga-DOTA-TATE and 18F-DOPA PET in patients with well-differentiated metastatic neuroendocrine tumours. AB - AIM: To compare the diagnostic impact of (68)Ga-DOTA-TATE and (18)F-DOPA PET in the diagnosis of well-differentiated metastatic neuroendocrine tumours (NET). METHODS: PET/CT using both (68)Ga-DOTA-TATE and (18)F-DOPA was performed in 25 patients with histologically proven metastatic NET (nine gut, five pancreas, six lung, one paranasal sinus, four with unknown primary). Analyses of PET examinations were patient-based (pathological uptake: yes/no), and based on tumour regions (primary tumour if present and metastases of liver, lung, bones and lymph nodes). The results were compared with the results of contrast enhanced CT, and with plasma serotonin levels, which were available in 24 of the 25 patients. RESULTS: Patient-based sensitivities were 96% for (68)Ga-DOTA-TATE PET and 56% for (18)F-DOPA PET. (68)Ga-DOTA-TATE PET delineated metastases in 54 of 55 positive metastatic tumour regions in contrast to 29 of 55 delineated by (18)F DOPA PET. Overall, (68)Ga-DOTA-TATE was superior to (18)F-DOPA in 13 patients (two patients showed fewer positive tumour regions with (18)F-DOPA PET). The results were comparable in 12 patients. In 13 of 24 patients, plasma serotonin levels were elevated, and 11 of these 13 patients showed pathological uptake of (18)F-DOPA. Of the 11 patients with normal levels of serotonin, 3 also showed positive (18)F-DOPA uptake. In patients positive for (18)F-DOPA uptake the maximum tumour SUVs were correlated with the levels of serotonin (r=0.66, p=0.01). CONCLUSION: In this study (68)Ga-DOTA-TATE PET proved clearly superior to (18)F-DOPA PET for detection and staging of NET. (18)F-DOPA uptake tended to be increased in those patients with elevated plasma serotonin. We conclude that (18)F-DOPA PET should be employed in patients with NET with negative (68)Ga-DOTA TATE PET and elevated plasma serotonin. PMID- 19137294 TI - Cerebral perfusion (HMPAO-SPECT) in patients with depression with cognitive impairment versus those with mild cognitive impairment and dementia of Alzheimer's type: a semiquantitative and automated evaluation. AB - PURPOSE: Comparative evaluation of regional brain perfusion measured by HMPAO SPECT of patients with mild cognitive impairment (MCI), dementia of Alzheimer's type (DAT) and depression with cognitive impairment (DCI). METHODS: A total of 736 patients were investigated because of suspected cognitive dysfunction. After exclusion of patients with other forms of dementia than DAT or relevant accompanying disorders, SPECT data from 149 MCI, 131 DAT and 127 DCI patients, and 123 controls without any cognitive impairment, were analysed. Relative cerebral blood flow of 34 anatomical regions was assessed with automated analysis software (BRASS). RESULTS: Calculation of global forebrain perfusion discriminated demented from nondemented patients. Compared to controls DCI patients showed hypoperfusion of the thalamus, lentiform nucleus and medial temporal cortex. MCI patients differed significantly from controls concerning perfusion in both hemispheric temporal and parietal areas, and in the (right hemispheric) posterior part of the cingulate gyrus. MCI and DCI patients differed in the parietal, temporal superior and right hemispheric cingulate gyrus posterior cortices. Global forebrain and regional perfusion was more extensively reduced in DAT patients and discriminated them from controls, and MCI and DCI patients. Frontal perfusion disturbance was only present in DAT patients. CONCLUSION: Automated analysis of HMPAO-SPECT data from MCI patients showed significant perfusion deficits in regions also involved in DAT patients, but ROC analysis demonstrated only moderate sensitivity and specificity for differentiating DAT patients from controls and DCI patients. Frontal hypoperfusion seems to correspond with conversion from MCI to DAT. Finally, the results in DCI patients again raise the question of depression as an early symptom of neurodegeneration. PMID- 19137295 TI - 18F-fluorodeoxyglucose and PET/CT for noninvasive study of exercise-induced glucose uptake in rat skeletal muscle and tendon. AB - PURPOSE: To investigate exercise-related glucose uptake in rat muscle and tendon using PET/CT and to study possible explanatory changes in gene expression for the glucose transporters (GLUT1 and GLUT4). METHODS: The sciatic nerve in eight Wistar rats was subjected to electrostimulation to cause unilateral isometric contractions of the calf muscle. (18)F-Fluorodeoxyglucose was administered and a PET/CT scan of the hindlimbs was performed. SUVs were calculated in both Achilles tendons and the triceps surae muscles. To exclude a spill-over effect the tendons and muscles from an ex vivo group of eight rats were cut out and scanned separately (distance>or=1 cm). RESULTS: Muscle contractions increased glucose uptake approximately sevenfold in muscles (p<0.001) and 36% in tendons (p<0.01). The ex vivo group confirmed the increase in glucose uptake in intact animals. GLUT1 and GLUT4 were expressed in both skeletal muscle and tendon, but no changes in mRNA levels could be detected. CONCLUSION: PET/CT can be used for studying glucose uptake in rat muscle and tendon in relation to muscle contractions; however, the increased uptake of glucose was not explained by changes in gene expression of GLUT1 and GLUT4. PMID- 19137296 TI - Tibialis posterior transfer by interosseous route for the correction of foot drop in leprosy. AB - This article summarises a prospective study to evaluate the long-term results produced by interosseous transfer of the tibialis posterior tendon for the correction of foot drop due to leprosy neuritis. The study was carried out in 120 feet in 69 patients. All patients had closed elongation of the tendo Achillis (ETA) before transfer of the bifurcated tibialis posterior tendon through the interosseous route to the tendons of tibialis anterior and peroneous tertius or brevis over the dorsum of feet. At final follow-up of average 24 months, all the patients with ETA had a significantly greater range of active dorsiflexion of more than 10 degrees above 90 degrees, which was not merely from the tenodesing effect. The results, in terms of improvement in gait and prevention of trophic changes, remained satisfactory. An interosseous route is preferred with split attachment to the tibialis anterior and to the peroneus brevis or tertius tendons. PMID- 19137297 TI - Allelic genotyping reveals a hierarchy of genomic alterations in mantle cell lymphoma associated to cell proliferation. AB - Mantle cell lymphoma (MCL) is a distinct subentity of non-Hodgkin lymphoma, characterized by the chromosomal translocation t(11;14)(q13;q32) leading to an overexpression of cyclin D1 in virtually all cases. However, additional cytogenetic aberrations are apparent in the vast majority of MCL. Applying LOH analysis in 52 MCL patient samples, we confirmed frequent alterations in 9p21 (28.6%) and p53 (28.9%) but also detected allelic losses in 1p21, 9q21, 13q13-14, 13q31-32, 17p13.1, and 17p13.3 in 28-45% of cases and allelic gains in 3q27-28 and 19p13.3 in 14-22% of cases. In addition, losses in the 2p23 and 7q22-35 genomic regions not previously described to be altered in MCL were identified in up to 20% of cases. Applying multivariate analysis, a cluster of genomic aberrations including 1p21, 3q27, 7q22-36, 6p24, 9p21, 9q31, and 16p12 alterations was identified which was closely associated to cell proliferation as determined by Ki67 immunostaining. This proliferation-dependent network of oncogenic alterations complements the previously identified proliferation expression signature described by RNA expression profiling in MCL. PMID- 19137298 TI - A rare combination of two inherited disorders in one patient: pyruvate kinase deficiency and hemochromatosis. PMID- 19137299 TI - Soluble transferrin receptor-ferritin index and estimated body iron in iron deficiency anemia in "select" chronic diseases. PMID- 19137300 TI - Identification of the het-r vegetative incompatibility gene of Podospora anserina as a member of the fast evolving HNWD gene family. AB - In fungi, vegetative incompatibility is a conspecific non-self recognition mechanism that restricts formation of viable heterokaryons when incompatible alleles of specific het loci interact. In Podospora anserina, three non-allelic incompatibility systems have been genetically defined involving interactions between het-c and het-d, het-c and het-e, het-r and het-v. het-d and het-e are paralogues belonging to the HNWD gene family that encode proteins of the STAND class. HET-D and HET-E proteins comprise an N-terminal HET effector domain, a central GTP binding site and a C-terminal WD repeat domain constituted of tandem repeats of highly conserved WD40 repeat units that define the specificity of alleles during incompatibility. The WD40 repeat units of the members of this HNWD family are undergoing concerted evolution. By combining genetic analysis and gain of function experiments, we demonstrate that an additional member of this family, HNWD2, corresponds to the het-r non-allelic incompatibility gene. As for het-d and het-e, allele specificity at the het-r locus is determined by the WD repeat domain. Natural isolates show allelic variation for het-r. PMID- 19137301 TI - Coil embolization of cavernous sinus in patients with direct and dural arteriovenous fistula. AB - To determine technical success and acute complication rates after endovascular coil packing of the cavernous sinus. Nineteen patients presented with either direct (13) or dural (6) arteriovenous fistula (AVF) and were treated by means of coil embolization of the cavernous sinus. The aim of treatment was complete obliteration of the fistula. In a retrospective study, the degree of obliteration, regression of symptoms as well as complication rates were evaluated. Initial complete obliteration was achieved in 12 patients, subtotal occlusion of the sinus in 6 and incomplete packing with major residual fistula in 1 of the patients. Retreatment was successfully performed in two patients with early recurrence of AVF. Follow-up showed complete occlusion rates in 16 and subtotal obliteration in 3 patients. Chemosis and exophthalmus regressed rapidly in all affected patients. Persistence of cranial nerve deficits was observed in 11 cases. Postinterventional thrombosis of the ophthalmic vein was the only major acute complication (n = 2). Coil embolization of the cavernous sinus in cases with AVF is a complex procedure that is technically feasible and safe in the majority of cases. Adequate anticoagulation is recommended to avoid thrombembolic complications. Long-term outcome has to be determined by further studies. PMID- 19137302 TI - Imaging in covert ectopic ACTH secretion: a CT pictorial review. AB - The syndrome of ectopic adrenocorticotrophin secretion (EAS) is rare and is due to excess adrenocorticotrophin (ACTH) production from a nonpituitary tumour. These tumours can be covert, where the tumours are not readily apparent, and very small making them challenging to image. It is clinically and biochemically difficult to distinguish between covert EAS and Cushing's disease. The first-line investigation in locating the source of ACTH production is computed tomography (CT). The aim of this pictorial review is to illustrate the likely covert sites and related imaging findings. We review the CT appearances of tumours resulting in covert EAS and the associated literature. The most common tumours were bronchial carcinoid tumours, which appear as small, well-defined, round or ovoid pulmonary lesions. Rarer causes included thymic carcinoids, gastrointestinal carcinoids and pancreatic neuroendocrine tumours. Awareness of the imaging characteristics will aid identification of the source of ACTH production and allow potentially curative surgical resection. PMID- 19137303 TI - A simple MRI protocol in patients with clinically suspected appendicitis: results in 138 patients and effect on outcome of appendectomy. AB - To establish the value of breathhold magnetic resonance imaging (MRI) in the diagnosis of acute appendicitis. Over a 14-month period, 138 patients clinically suspected of having appendicitis were evaluated prospectively with MRI and comprised the study group. Fast turbo spin-echo breathhold T1, T2 and T2 fat suppression sequences were used in coronal and axial planes. The imaging results were recorded separately and subsequently correlated with clinical, radiological and histopathological follow-up. The effect of imaging strategies in patients suspected of appendicitis on hospital resources was calculated. Sixty-two of the 138 patients had a histopathologically proven appendicitis. MRI determined appendicitis in 63 patients, with one examination being false positive. The resulting sensitivity and specificity were 100% and 99%, respectively. MRI showed an alternative diagnosis in 41 of the 75 remaining patients. In 22 of the remaining 34 patients, a normal appendix was depicted with MRI. In two patients, where MRI showed no appendicitis, an alternative diagnosis or normal appendix, an unnecessary appendectomy was performed. The overall effect of using MRI in patients suspected of appendicitis on the use of hospital resources could have been a net saving between 55,746 euros and 72,534 euros. MRI has a high accuracy in detecting and excluding appendicitis, an alternative diagnosis or showing the normal appendix, and can be a valuable and cost-effective tool in the workup of patients clinically suspected of having appendicitis. PMID- 19137304 TI - Diffuse optical tomography of the breast: preliminary findings of a new prototype and comparison with magnetic resonance imaging. AB - This paper presents an evaluation of a prototype diffuse optical tomography (DOT) system. Seventeen women with 18 breast lesions (10 invasive carcinomas, 2 fibroadenomas, and 6 benign cysts; diameters 13-54 mm) were evaluated with DOT and magnetic resonance imaging (MRI). A substantial fraction of the original 36 recruited patients could not be examined using this prototype due to technical problems. A region of interest (ROI) was drawn at the lesion position as derived from MRI and at the mirror image site in the contralateral healthy breast. ROIs were assessed quantitatively and qualitatively by two observers independently in two separate readings. Intra- and interobserver agreements were calculated using kappa statistics (k) and intraclass correlation coefficients (ICCs). Discriminatory values for presence of malignancy were determined by receiver operating characteristic (ROC) analyses. Intraobserver agreements were excellent (k 0.88 and 0.88; ICC 0.978 and 0.987), interobserver agreements were good to excellent (k 0.77-0.95; ICC 0.96-0.98). Discriminatory values for presence of malignancy were 0.92-0.93 and 0.97-0.99 for quantitative and qualitative ROC analysis, respectively. This DOT system has the potential to discriminate malignant from benign breast tissue in a reproducible qualitative and quantitative manner. Important technical improvements are required before this technique is ready for clinical application. PMID- 19137305 TI - 64-Slice spiral computed tomography of the coronary arteries: dose reduction using an optimized imaging protocol including individual weight-adaptation of voltage and current-time product. AB - Radiation dose and image quality were compared between a standard protocol (40 patients, group A) and a weight-adapted protocol of voltage and current-time product (44 patients, group B) using 64-slice coronary multidetector computed tomography (MDCT). Effective dose estimate was lower by 37% in all patients of group B (9.2 +/- 2.5 mSv) compared with group A (14.6 +/- 2.3 mSv, P < 0.0001). Group B patients with a small body mass index (BMI) benefited most with a dose reduction of 53% (6.7 +/- 1.5 mSv in group B versus 14.1 +/- 1.8 mSv in group A, P < 0.0001). Moderate reductions of 32% and 20% were achieved for patients with a medium and large BMI, respectively. Reduction in radiation dose did not affect the image quality as assessed by image noise, signal-to-noise ratios, and number of coronary segments with good diagnostic image quality. Individual weight adaptation of voltage and current-time product significantly reduces the radiation dose without loss of image quality. PMID- 19137306 TI - Diagnosis of temporomandibular dysfunction syndrome--image quality at 1.5 and 3.0 Tesla magnetic resonance imaging. AB - The purpose of this study was to examine the differences in expert ratings of quality of magnetic resonance images (MRI) of the temporomandibular joint in 24 patients with suspected anterior disc displacement examined in randomized order at 1.5 and 3.0 T. Parasagittal (closed and opened mouth) and paracoronal sections were performed with a surface coil. Two experienced examiners blinded to patient status and type of MRI diagnosed the images according to position of condyle, position and changes in the signal, and disc shape. In addition, perceptibility of position and disc shape were assessed. A highly significant difference in the perceptibility of disc shape (P < 0.001) and position (P < 0.001) was obtained. With comparable examination sequences and identical resolution, the 3.0 T MRI of the temporomandibular joint increases the perceptibility of joint structures. PMID- 19137307 TI - Animal models of cancer in interventional radiology. AB - Animal models will play an increasingly important role in oncology research, especially for solid tumours such as hepatocellular carcinoma that are resistant to chemotherapy. Many models have been used, but there is a need for increased awareness of the limitations of these models and also a need for guidance for future model development. PMID- 19137308 TI - Magnetic targeting of iron-oxide-labeled fluorescent hepatoma cells to the liver. AB - The purpose of this study was to determine whether an external magnet field can induce preferential trafficking of magnetically labeled Huh7 hepatoma cells to the liver following liver cell transplantation. Huh7 hepatoma cells were labeled with anionic magnetic nanoparticles (AMNP) and tagged with a fluorescent membrane marker (PKH67). Iron-uptake was measured by magnetophoresis. Twenty C57Bl6 mice received an intrasplenic injection of 2 x 10(6) labeled cells. An external magnet (0.29 T; 25 T/m) was placed over the liver of 13 randomly selected animals (magnet group), while the remaining 7 animals served as controls. MRI (1.5 T) and confocal fluorescence microscopy (CFM) were performed 10 days post transplantation. The presence and location of labeled cells within the livers were compared in the magnet group and controls, and confronted with histological analysis representing the standard of reference. Mean iron content per cell was 6 pg. Based on histology, labeled cells were more frequently present within recipient livers in the magnet group (p < 0.01) where their distribution was preferentially peri-vascular (p < 0.05). MRI and CFM gave similar results for the overall detection of transplanted cells (kappa = 0.828) and for the identification of peri-vascular cells (kappa = 0.78). Application of an external magnet can modify the trafficking of transplanted cells, especially by promoting the formation of perivascular aggregates. PMID- 19137309 TI - Isotropic 3D fast spin-echo imaging versus standard 2D imaging at 3.0 T of the knee--image quality and diagnostic performance. AB - The objective of this study was to compare a newly developed fat-saturated intermediate-weighted (IM-w) 3D fast spin-echo (FSE) sequence with standard 2D IM w FSE sequences regarding image quality and diagnostic performance in assessing abnormal findings of the knee. MR imaging was performed at 3.0 T in 50 patients. Images were assessed independently by three radiologists. Image quality was rated significantly higher (p < 0.05) for the 2D versus the 3D FSE sequences. Sensitivity for cartilage lesions was slightly higher for the 3D sequence, but specificity was lower. Low contrast objects were better visualized with 2D sequences, while high contrast objects were better shown with the 3D sequence. Confidence scores were higher for 2D than for 3D sequences, but differences were not significant. In conclusion, isotropic 3D FSE IM-w imaging may enhance standard knee MRI by increased visualization of high contrast lesions; however, 3D FSE image quality was lower. PMID- 19137310 TI - Five-year survival in 309 patients with colorectal liver metastases treated with radiofrequency ablation. AB - There is little published long-term survival data for patients with colorectal liver metastases treated with radiofrequency ablation (RFA). We present a multivariate analysis of 5-year survival in 309 patients (198 male, aged 64 (24 92)) treated at 617 sessions. Our standard protocol used internally cooled electrodes introduced percutaneously under combined US and CT guidance/monitoring. The number and size of liver metastases, the presence and location of extrahepatic disease, primary resection, clinical, chemotherapy and follow-up data were recorded. Data analysis was performed using SPSS v.10. On multivariate analysis, significant survival factors were the presence of extrahepatic disease (p < 0.001) and liver tumour volume (p = 0.001). For 123 patients with five or less metastases of 5 cm or less maximum diameter and no extrahepatic disease median survival was 46 and 36 months from liver metastasis diagnosis and ablation, respectively; corresponding 3- and 5-year survival rates were 63%, 34% and 49%, 24%. Sixty-nine patients had three or less tumours of below 3.5 cm in diameter and their 5-year survival from ablation was 33%. There were 23/617(3.7%) local complications requiring intervention. Five-year survival of 24-33% post ablation in selected patients is superior to any published chemotherapy data and approaches the results of liver resection. PMID- 19137311 TI - Initial experience with a chest pain protocol using 320-slice volume MDCT. AB - We sought to determine the feasibility and image quality of 320-slice volume computed tomography (CT) angiography for the evaluation of patients with acute chest pain. Thirty consecutive patients (11 female, 19 male, mean age 63.2 +/- 14.2 years) with noncritical, acute chest pain underwent 320-slice CT using a protocol consisting of a nonspiral, nongated CT of the entire chest, followed by a nonspiral, electrocardiography-gated CT study of the heart. Data were acquired following a biphasic intravenous injection of 90 ml iodinated contrast agent. Vessel attenuation values of different thoracic vascular territories were recorded, and image quality scored on a five-point scale by two readers. Mean attenuation was 467 +/- 69 HU in the ascending aorta, 334 +/- 52 HU in the aortic arch, 455 +/- 71 HU in the descending aorta, 492 +/- 94 HU in the pulmonary trunk, and 416 +/- 63 HU and 436 +/- 62 HU in the right and left coronary artery, respectively. Radiation exposure estimates ranged between 7 and 14 mSv. The CT protocol investigated enabled imaging of the thoracic aorta, coronary and pulmonary arteries with an excellent diagnostic quality for chest pain triage in all patients. This result was achieved with less contrast material and reduced radiation exposure compared with previously investigated imaging protocols. PMID- 19137312 TI - Dynamic contrast-enhanced CT imaging of hepatocellular carcinoma in cirrhosis: feasibility of a prolonged dual-phase imaging protocol with tracer kinetics modeling. AB - Dynamic contrast-enhanced (DCE) CT imaging of four patients with hepatocellular carcinoma (HCC) was performed using a dual-phase imaging protocol designed with initial rapid dynamic imaging to capture the initial increase in contrast medium enhancement in order to assess perfusion, followed by a delayed imaging phase with progressively longer intervals to monitor subsequent tissue enhancement behaviour in order to assess tissue permeability. The DCE CT images were analysed using a dual-input two-compartment distributed parameter model to yield separate estimates for blood flow and permeability, as well as fractional intravascular and extravascular volumes. The HCCs and surrounding cirrhotic liver tissues were found to exhibit enhancement curves that can be appropriately described by two distinct compartments separated by a semipermeable barrier. Early contrast arrival was also found for HCC as compared with background liver. These findings are consistent with the current understanding of sinusoidal capillarization and hepatocarcinogenesis. PMID- 19137313 TI - Does the echogenicity of focal liver lesions on baseline gray-scale ultrasound interfere with the diagnostic performance of contrast-enhanced ultrasound? AB - The objective was to evaluate whether the echogenicity of focal liver lesions (FLLs) on baseline gray-scale ultrasound (US) interferes with the diagnostic performance of contrast-enhanced US (CEUS) for small FLLs. Three-hundred and eighty-eight patients were examined by real-time CEUS using a sulfur hexafluoride filled microbubble contrast agent. The images of 114 hyperechoic lesions, 30 isoechoic lesions and 244 hypoechoic lesions were reviewed by two blinded independent readers. A five-point confidence level was used to discriminate malignant from benign lesions, and specific diagnoses were made. The diagnostic performances were evaluated by receiver-operating characteristic (ROC) analysis. The diagnostic performances of CEUS on hyperechoic lesions in terms of the areas (Az) under the ROC curve were 0.987 (reader 1) and 0.981 (reader 2), and were 0.987 (reader 1) and 0.984 (reader 2) for iso- and hypoechoic lesions, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 87.0-95.9%, 93.1-100%, 88.6-100%, 70.0-97.1% and 90.0-95.1%, respectively. The echogenicity of FLLs on baseline gray-scale US does not appear to interfere with the diagnostic ability of CEUS for small FLLs. PMID- 19137315 TI - Surface structure and frictional properties of the skin of the Amazon tree boa Corallus hortulanus (Squamata, Boidae). AB - The legless locomotion of snakes requires specific adaptations of their ventral scales to maintain friction force in different directions. The skin microornamentation of the snake Corallus hortulanus was studied by means of scanning electron microscopy and the friction properties of the skin were tested on substrates of different roughness. Skin samples from various parts of the body (dorsal, lateral, ventral) were compared. Dorsal and lateral scales showed similar, net-like microornamentation and similar friction coefficients. Average friction coefficients for dorsal and lateral scales on the epoxy resin surfaces were 0.331 and 0.323, respectively. In contrast, ventral scales possess ridges running parallel to the longitudinal body axis. They demonstrated a significantly lower friction coefficient compared to both dorsal and lateral scales (0.191 on average). In addition, ventral scales showed frictional anisotropy comparing longitudinal and perpendicular direction of the ridges. This study clearly demonstrates that different skin microstructure is responsible for different frictional properties in different body regions. PMID- 19137314 TI - Variability of bronchial measurements obtained by sequential CT using two computer-based methods. AB - This study aimed to evaluate the variability of lumen (LA) and wall area (WA) measurements obtained on two successive MDCT acquisitions using energy-driven contour estimation (EDCE) and full width at half maximum (FWHM) approaches. Both methods were applied to a database of segmental and subsegmental bronchi with LA > 4 mm(2) containing 42 bronchial segments of 10 successive slices that best matched on each acquisition. For both methods, the 95% confidence interval between repeated MDCT was between -1.59 and 1.5 mm(2) for LA, and -3.31 and 2.96 mm(2) for WA. The values of the coefficient of measurement variation (CV(10), i.e., percentage ratio of the standard deviation obtained from the 10 successive slices to their mean value) were strongly correlated between repeated MDCT data acquisitions (r > 0.72; p < 0.0001). Compared with FWHM, LA values obtained using EDCE were higher for LA < 15 mm(2), whereas WA values were lower for bronchi with WA < 13 mm(2); no systematic EDCE underestimation or overestimation was observed for thicker-walled bronchi. In conclusion, variability between CT examinations and assessment techniques may impair measurements. Therefore, new parameters such as CV(10) need to be investigated to study bronchial remodeling. Finally, EDCE and FWHM are not interchangeable in longitudinal studies. PMID- 19137316 TI - Tight turns in stick insects. AB - We investigated insects Carausius morosus walking whilst hanging upside down along a narrow 3 mm horizontal beam. At the end of the beam, the animal takes a 180 degrees turn. This is a difficult situation because substrate area is small and moves relative to the body during the turn. We investigated how leg movements are organised during this turn. A non-contact of either front leg appears to indicate the end of the beam. However, a turn can only begin if the hind legs stand in an appropriate position relative to each other; the outer hind leg must not be placed posterior to the inner hind leg. When starting the turn, both front legs are lifted and usually held in a relatively stable position and then the inner middle leg performs a swing-and-search movement: The leg begins a swing, which is continued by a searching movement to the side and to the rear, and eventually grasps the beam. At the same time the body is turned usually being supported by the outer middle leg and both hind legs. Then front legs followed by the outer middle leg reach the beam. A scheme describing the turns based on a few simple behavioural elements is proposed. PMID- 19137317 TI - Lateral line stimulation patterns and prey orienting behavior in the Lake Michigan mottled sculpin (Cottus bairdi). AB - Information contained in the spatial excitation pattern along arrayed sensors in the lateral line system of Lake Michigan mottled sculpin, as well as other surface-feeding fish and amphibians, is thought to play a fundamental role in guiding prey-orienting behaviors. However, the way in which prey location is encoded by the excitation pattern and used by the nervous system to direct orienting behaviors is largely unknown. In this study, we test the hypothesis that mottled sculpin use excitation peaks (local 'hot spots') to determine the somatotopic location of an artificial prey (vibrating sphere/dipole source) along the body surface. Dipole orientation (axis of sphere vibration re: long axis of the fish) is manipulated to produce excitatory peaks in different body locations without changing the actual sphere location. Our results show that orienting accuracy is largely independent of source orientation, but not source distance and that turning directions are not guided by local hot spots in the somatotopic activation pattern of the lateral line. PMID- 19137318 TI - Differential effects of octopamine and tyramine on the central pattern generator for Manduca flight. AB - The biogenic amine, octopamine, modulates a variety of aspects of insect motor behavior, including direct action on the flight central pattern generator. A number of recent studies demonstrate that tyramine, the biological precursor of octopamine, also affects invertebrate locomotor behaviors, including insect flight. However, it is not clear whether the central pattern generating networks are directly affected by both amines, octopamine and tyramine. In this study, we tested whether tyramine affected the central pattern generator for flight in the moth, Manduca sexta. Fictive flight was induced in an isolated ventral nerve cord preparation by bath application of the octopamine agonist, chlordimeform, to test potential effects of tyramine on the flight central pattern generator by pharmacological manipulations. The results demonstrate that octopamine but not tyramine is sufficient to induce fictive flight in the isolated ventral nerve cord. During chlordimeform induced fictive flight, bath application of tyramine selectively increases synaptic drive to depressor motoneurons, increases the number of depressor spikes during each cycle and decreases the depressor phase. Conversely, blocking tyramine receptors selectively reduces depressor motoneuron activity, but does not affect cycle by cycle elevator motoneuron spiking. Therefore, octopamine and tyramine exert distinct effects on the flight central pattern generating network. PMID- 19137319 TI - Lake char (Salvelinus namaycush) olfactory neurons are highly sensitive and specific to bile acids. AB - Bile acids have been implicated as chemical signals in spawning behaviour of lake char (Salvelinus namaycush). In this study, we investigated olfactory responses of lake char to bile acids by using the electro-olfactogram recording. Lake char detected 9 out of 38 bile acids tested at thresholds 0.02-0.5 nM. The most stimulatory included chenodeoxycholic acid, cholic acid, taurochenodeoxycholic acid, taurocholic acid, and taurolithocholic acid 3alpha-sulphate. Structure activity analysis indicated that substituents in the side chain or hydroxyl sulphation were determinant elements for the recognition of individual bile acid receptors, while the position and orientation of hydroxyls or the type of amidation were important for effective stimulation. Three distinct types of concentration-response relationships were found, representing free, taurine- or glycine-amidated, and 3alpha-sulphated bile acids. Cross-adaptation and binary mixture experiments revealed the presence of multiple olfactory receptors for bile acids. Lake char were also capable of detecting petromyzonol sulphate at 1 nM, possibly via its own receptors. Our study further showed that the olfactory responses to bile acids were independent of those of known odorants including amino acids, prostaglandins and gonadal steroids. We conclude that lake char possess multiple olfactory receptors capable of discriminating bile acids produced and released by conspecifics. PMID- 19137320 TI - Topical anal fissure treatment: placebo-controlled study of mononitrate and trinitrate therapies. AB - AIM: The present study aims to evaluate and compare the efficacy of two nitrate gels, containing isosorbide-5-mononitrate (ISMN) or glyceryl trinitrate (GTN), in the therapy of chronic anal fissure. MATERIALS AND METHODS: The patients were randomly assigned to three groups: 0.1% ISMN gel (21 patients), 0.1% GTN gel (21 patients) and a placebo group (ten patients). The ethic committee of our hospital approved the protocol and informed consent was obtained from all participants. All patients underwent clinical examination, visual inspection of the fissure and anal manometry prior to and after therapy. RESULTS: The chronic anal fissure was completely healed in 71% of the patients treated with ISMN, 67% with GTN and in 30% from the placebo group. One patient in the ISMN group reported mild headache. Three patients in the GTN group had anal burning. CONCLUSION: Both topical nitrate treatments (ISMN and GTN) were effective for chronic anal fissures. The reduction of the anal pressure was slightly higher after ISMN treatment (28%) than the treatment with GTN (23%). However, the statistical difference was not significant (p>0.05). PMID- 19137321 TI - Rhombencephalosynapsis with and without associated malformations. PMID- 19137322 TI - Nomenclature discrepancies in the HPRTB short tandem repeat. PMID- 19137323 TI - A comparison of critical force and electromyographic fatigue threshold for isometric muscle actions of the forearm flexors. AB - The purpose of this study was twofold: (1) to determine if the mathematical model used for estimating the EMG(FT) during cycle ergometry was applicable to isometric muscle actions; and (2) to compare the mean torque level from the CF test to that of the EMG(FT) test. The CF was defined as the slope coefficient of the linear relationship between total "isometric work" (W (lim) in N m s) and time to exhaustion (T (lim)). The EMG(FT) was defined as the y-intercept of the isometric torque versus EMG fatigue curve slope coefficient relationship. There was a significant (p < 0.05) mean difference between CF (6.6 +/- 3.2 N m) and EMG(FT) (10.9 +/- 4.7 N m). The results of the present study suggested that, during isometric muscle actions of the forearm flexors, fatigue thresholds estimated from the W (lim) versus T (lim) relationship (CF) are different from those estimated from electromyographic fatigue curves (EMG(FT)). PMID- 19137324 TI - Paediatric intracranial empyema: differences according to age. AB - No recent studies are available which consider the epidemiology and outcome of paediatric intracranial empyema (PICE). We retrospectively studied all PICE cases admitted in our institution from 1993 to 2006. Outcome was assessed using the Glasgow Outcome Scale (GOS) at 24 months. Aetiology, clinical features, therapeutic considerations and risk factors of poor outcome were analysed according to age. Data from 38 patients were studied; 33/38 presented with subdural empyema (SDE) and 5/38 with extradural empyema (EDE); 10/38 were infants <1 year of age with SDE, all related to bacterial meningitis; 28/38 were children, with 23/28 showing SDE and 5/28 EDE. Oto-sinogenic infections were the main causes in children. All infants recovered completely as did children with EDE. However, two out of 23 children with SDE had permanent neurological deficit, already detected on admission, and one out of 23 died. Thirty-three out of 38 were operated; 16 of which underwent multiple surgical procedure because of recurrence. Burr hole was performed in six infants and craniotomy in one, while 21/23 children underwent burr hole or craniotomy. Burr hole was more often associated with recurrence. In children with SDE, factors associated with poor outcome were neurological deficit (p = 0.002) and cerebral herniation on CT scan (p = 0.02) on admission. In this study, we gained further insights into modern epidemiology of PICE by highlighting age-related aetiology, symptoms, treatment strategy, and outcome differences. Meningitis was the main aetiology in the infants and sinusitis was prevalent in children. Finally, early diagnosis by neuro-imaging investigations and timely and appropriate multidisciplinary treatment may offer the best chance of recovery. PMID- 19137325 TI - Broken heart syndrome in a 17-year-old girl. AB - Broken heart syndrome is characterized by transient left ventricular dysfunction with chest pain, electrocardiographic changes, and release of myocardial enzymes mimicking acute myocardial infarction. Physical and emotional stress usually precede the onset of symptoms. This dysfunction, known as takotsubo cardiomyopathy (TC), has been observed predominantly in women older than 60 years of age but very rarely reported in younger adults or children. Here we describe a 17-year-old girl with TC. PMID- 19137327 TI - Transplacental transmission of Toxoplasma gondii in reinfected pregnant female canines. AB - Twelve pregnant female canines, naturally infected with Toxoplasma gondii, were reinfected with T. gondii: three (GI) received tachyzoites subcutaneously (1.0 x 107), three (GII) were orally inoculated with oocysts (1.5 x 104), and six (GIII) were kept as a nonreinfected control group. All the reinfected female canines (GI and GII) miscarried or presented fetal death, while only one GIII female presented a stillborn in a litter of four pups (P < 0.01). Fever, lymphoadenopathy, miscarriage, and fetal death were the main clinical alterations observed. The highest serological titers detected through the indirect fluorescence antibody test (IFAT) were 1,024 (GI) and 4,096 (GII). In group III, the titers ranged between 64 and 256. By bioassays in mice, T. gondii was isolated in 17 organs of the reinfected adult canines, in 11 of the control group, and in 20 of the neonates. Positive immunostaining of cysts and/or tachyzoites were observed in 26 canine tissues (14 from GI and GII and ten from GIII). The agent was detected by immunohistochemistry in the encephalon of a neonate and in the spinal cord of a stillborn, thus, confirming that T. gondii infected canine fetuses, provoking miscarriages, even in bitches that presented primoinfection. PMID- 19137328 TI - Photosynthetic performance in Sphagnum transplanted along a latitudinal nitrogen deposition gradient. AB - Increased N deposition in Europe has affected mire ecosystems. However, knowledge on the physiological responses is poor. We measured photosynthetic responses to increasing N deposition in two peatmoss species (Sphagnum balticum and Sphagnum fuscum) from a 3-year, north-south transplant experiment in northern Europe, covering a latitudinal N deposition gradient ranging from 0.28 g N m(-2) year(-1) in the north, to 1.49 g N m(-2) year(-1) in the south. The maximum photosynthetic rate (NP(max)) increased southwards, and was mainly explained by tissue N concentration, secondly by allocation of N to the photosynthesis, and to a lesser degree by modified photosystem II activity (variable fluorescence/maximum fluorescence yield). Although climatic factors may have contributed, these results were most likely attributable to an increase in N deposition southwards. For S. fuscum, photosynthetic rate continued to increase up to a deposition level of 1.49 g N m(-2) year(-1), but for S. balticum it seemed to level out at 1.14 g N m(-2) year(-1). The results for S. balticum suggested that transplants from different origin (with low or intermediate N deposition) respond differently to high N deposition. This indicates that Sphagnum species may be able to adapt or physiologically adjust to high N deposition. Our results also suggest that S. balticum might be more sensitive to N deposition than S. fuscum. Surprisingly, NP(max) was not (S. balticum), or only weakly (S. fuscum) correlated with biomass production, indicating that production is to a great extent is governed by factors other than the photosynthetic capacity. PMID- 19137329 TI - Measuring the effects of heat wave episodes on the human body's thermal balance. AB - During the peak of an extensive heat wave episode on 23-25 July 2007, simultaneous thermophysiological measurements were made in two non-acclimated healthy adults of different sex in a suburban area of Greater Athens, Greece. Based on experimental measurements of mean skin temperature and metabolic heat production, heat fluxes to and from the human body were calculated, and the biometeorological index heat load (HL) produced was determined according to the heat balance equation. Comparing experimental values with those derived from theoretical estimates revealed a great heat stress for both individuals, especially the male, while theoretical values underestimated heat stress. The study also revealed that thermophysiological factors, such as mean skin temperature and metabolic heat production, play an important role in determining heat fluxes patterns in the heat balance equation. The theoretical values of mean skin temperature as derived from an empirical equation may not be appropriate to describe the changes that take place in a non-acclimated individual. Furthermore, the changes in metabolic heat production were significant even for standard activity. PMID- 19137330 TI - Structural characterization of a novel Ca2+-binding protein from Entamoeba histolytica: structural basis for the observed functional differences with its isoform. AB - A novel Ca(2+)-binding protein (EhCaBP2) was identified from the protozoan parasite Entamoeba histolytica. EhCaBP2 has 79% sequence identity with calcium binding protein EhCaBP1. The 3D structure of EhCaBP2 was determined using multidimensional nuclear magnetic resonance spectroscopic techniques. The study reveals that the protein consists of two globular domains connected by a short flexible linker region of four residues. On comparison of the 3D structure and dynamics of EhCaBP2 with those of EhCaBP1, it is found that they vary significantly in their N-terminal domains and interdomain linker. Immunofluorescence localization experiments revealed that EhCaBP1 and EhCaBP2 may not carry out similar functions, as their cellular distribution patterns are not the same. The functional differences between the two isoforms are explained on the basis of results obtained from the structural studies. The structural variation in the interdomain linker region and the formation of functionally important hydrophobic clefts in different regions of EhCaBP1 and EhCaBP2 provide interesting insights into the differences in the functionality of these two isoforms. PMID- 19137331 TI - The oral cavity--a key system to understand substratum-dependent bioadhesion on solid surfaces in man. AB - One of the greatest challenges in life sciences and biomaterials research is adhesion of biomolecules and bacteria to solid surfaces in aqueous solutions. An example concerning everybody is biofilm formation in the oral cavity on dental materials and dental hard substances, respectively. The main characteristics typical for any bioadhesion can be observed excellently in the oral cavity. Initially, a proteinaceous layer termed pellicle is formed. It mediates the interactions between solid substrata, oral fluids and microorganisms. Numerous different materials with differing physico-chemical properties and possible impact on the acquired pellicle are present in the oral cavity such as enamel, dentine, restorative materials or dental implants. Despite the fact that in vitro studies demonstrate considerable differences of experimental pellicles formed on these materials, the in situ pellicles seem to be relatively similar and level off the different properties of the underlying substrates. However, the bacterial colonisation of pellicle-coated surfaces under in vivo conditions differs considerably. Long-range forces and detachment of biofilm layers may account for this phenomenon despite the masking effect of the pellicle. Accordingly, low energy surfaces are desirable for restorative materials exposed to the oral cavity to minimise bacterial adhesion. The oral cavity is an easy accessible in vivo model for understanding bioadhesion and for investigation of protein-surface interactions noninvasively. For evaluation of biofilm formation on dental materials, in situ or in vivo studies are preferable. PMID- 19137332 TI - Influence of different cusp coverage methods for the extension of ceramic inlays on marginal integrity and enamel crack formation in vitro. AB - No information is available to date about cusp design of thin (1.0 mm) non functional cusps and its influence upon (1) marginal integrity of ceramic inlays (CI) and partial ceramic crowns (PCC) and (2) crack formation of dental tissues. The aim of this in vitro study was to investigate the effect of cusp coverage of thin non-functional cusps on marginal integrity and enamel crack formation. CI and PCC preparations were performed on extracted human molars. Non-functional cusps were adjusted to 1.0-mm wall thickness and 1.0-mm wall thickness with horizontal reduction of about 2.0 mm. Ceramic restorations (Vita Mark II, Cerec3 System) were adhesively luted with Excite/Variolink II. The specimens were exposed to thermocycling and central mechanical loading. Marginal integrity was assessed by evaluating dye penetration after thermal cycling and mechanical loading. Enamel cracks were documented under a reflective-light microscope. The data were statistically analysed with the Mann-Whitney U test, the Fishers exact test (alpha = 0.05) and the error rates method. PCC with horizontal reduction of non-functional cusps showed statistically significant less microleakage than PCC without such a cusp coverage. Preparation designs with horizontal reduction of non-functional cusps showed a tendency to less enamel crack formation than preparation designs without cusp coverage. Thin non-functional cusp walls of adhesively bonded restorations should be completely covered or reduced to avoid enamel cracks and marginal deficiency. PMID- 19137334 TI - High cell density production of Deinococcus radiodurans under optimized conditions. AB - Deinococcus radiodurans is a bacterium being investigated for mechanisms of extreme radiation resistance and for bioremediation of environmental radioactive waste sites. In both fundamental and applied research settings, methods for large scale production of D. radiodurans are needed. In this study, a systematic investigation was carried out to optimize D. radiodurans production at the 20-L fermentor scale. In defined medium, the phosphate buffer typically used was found to be inhibitory to D. radiodurans growth, and caused cell aggregation. Substitution of HEPES and MOPS buffers for phosphate buffer improved D. radiodurans growth characteristics. Several antifoaming agents were investigated to support large-scale production with submerged aeration, and the defoamer KFO 673 was chosen based on its ability to prevent foaming without affecting D. radiodurans growth. The conventional undefined rich medium tryptone/glucose/yeast extract (TGY) maximally supported D. radiodurans growth to an OD(600) of 10. Using a 'design of experiments' approach, we found glucose, Mg and Mn to be critical in supporting high-density growth of D. radiodurans. The optimal pH and temperature for D. radiodurans growth in large-scale preparations were 7.0 and 37 degrees C, respectively. Growth was carried out in a 20-L fermentor using the newly developed media under the optimal conditions. With addition of 10 g/L glucose, 0.5 g/L MgSO(4) . 7H(2)O, 5 microM MnCl(2) into TGY media, an OD(600) of 40 was achieved. PMID- 19137333 TI - Modeling the noncovalent interactions at the metabolite binding site in purine riboswitches. AB - We present gas phase quantum chemical studies on the metabolite binding interactions in two important purine riboswitches, the adenine and guanine riboswitches, at the B3LYP/6-31G(d,p) level of theory. In order to gain insights into the strucutral basis of their discriminative abilities of regulating gene expression, the structural properties and binding energies for the gas phase optimized geometries of the metabolite bound binding pocket are analyzed and compared with their respective crystal geometries. Kitaura-Morokuma analysis has been carried out to calculate and decompose the interaction energy into various components. NBO and AIM analysis has been carried out to understand the strength and nature of binding of the individual aptamer bases with their respective purine metabolites. The Y74 base, U in case of adenine riboswitch and C in case of guanine riboswitch constitutes the only differentiating element between the two binding pockets. As expected, with W:W cis G:C74 interaction contributing more than 50% of the total binding energy, the interaction energy for metabolite binding as calculated for guanine (-46.43 Kcal/mol) is nearly double compared to the corresponding value for that of adenine (-24.73 Kcal/mol) in the crystal context. Variations in the optimized geometries for different models and comparison of relative contribution to metabolite binding involving four conserved bases reveal the possible role of U47:U51 W:H trans pair in the conformational transition of the riboswitch from the metabolite free to metabolite bound state. Our results are also indicative of significant contributions from stacking and magnesium ion interactions toward cooperativity effects in metabolite recognition. PMID- 19137335 TI - DNA plasmid production in different host strains of Escherichia coli. AB - We compared plasmid DNA production in 13 strains of Escherichia coli in shake flasks using media containing glucose or glycerol. DNA yield from either carbon source showed small correlation with maximum growth rate. Three strains, SCS1-L, BL21 and MC4100, were selected for a controlled exponential fed-batch process at a growth rate of 0.14 h(-1) to an optical density of about 70, followed by a four hour heat treatment. Prior to heat treatment, SCS1-L generated 15.4 mg DNA/g, BL21 generated 11.0 mg DNA/g and MC4100 generated 7.9 mg DNA/g, while after heat treatment the strains attained DNA yields, respectively, of 18.0, 15.0 and 6.8 mg/g. The strains also varied in their percentage of supercoiled DNA after heat treatment, with SCS1-L averaging 66% supercoiled, BL21 17% and MC4100 40%. We further investigated the two strains that yielded the highest percentage of supercoiled DNA (SCS1-L and MC4100) at a higher growth rate of 0.28 h(-1). At this condition, a slightly lower DNA yield was generated faster, and the percentage of supercoiled DNA increased. Heat treatment improved DNA yield, and surprisingly did so to a greater extent at the higher growth rate. As a consequence of these factors, higher growth rates might be advantageous for DNA production. PMID- 19137336 TI - Biosolutions to the energy problem. AB - We are in an energy crisis caused by years of neglect to alternative energy sources. There are many possible solutions and a number of these are based on microorganisms. These include bioethanol, biobutanol, biodiesel, biohydrocarbons, methane, methanol, electricity-generating microbial fuel cells, and production of hydrogen via photosynthetic microbes. In this review, I will focus on the first four possibilities. PMID- 19137337 TI - Modeling and experimental studies on intermittent starch feeding and citrate addition in simultaneous saccharification and fermentation of starch to flavor compounds. AB - Simultaneous saccharification and fermentation (SSF) is a combined process of saccharification of a renewable bioresource and fermentation process to produce products, such as lactic acid and ethanol. Recently, SSF has been extensively used to convert various sources of cellulose and starch into fermentative products. Here, we present a study on production of buttery flavors, namely diacetyl and acetoin, by growing Lactobacillus rhamnosus on a starch medium containing the enzyme glucoamylase. We further develop a structured kinetics for the SSF process, which includes enzyme and growth kinetics. The model was used to simulate the effect of pH and temperature on the SSF process so as to obtain optimum operating conditions. The model was experimentally verified by conducting SSF using an initial starch concentration of 100 g/L. The study demonstrated that the developed kinetic was able to suggest strategies for improved productivities. The developed model was able to accurately predict the enhanced productivity of flavors in a three stage process with intermittent addition of starch. Experimental and simulations demonstrated that citrate addition can also lead to enhanced productivity of flavors. The developed optimal model for SSF was able to capture the dynamics of SSF in batch mode as well as in a three stage process. The structured kinetics was also able to quantify the effect of multiple substrates present in the medium. The study demonstrated that structured kinetic models can be used in the future for design and optimization of SSF as a batch or a fed-batch process. PMID- 19137338 TI - Influence of growth conditions on the nisin production of bioengineered Lactococcus lactis strains. AB - Nisin production of three bioengineered strains, (LAC338, LAC339 and LAC340) with immunity (nisFEG) and/or regulation (nisRK) genes of nisin biosynthesis on plasmids in the Lactococcus lactis LL27 nisin producer, was evaluated under pH controlled and pH-uncontrolled batch fermentations. Optimization studies showed that fructose and yeast extract yielded the highest nisin activity. The strains LAC338, LAC339, and LAC340 produced 24, 45, and 44% more nisin, respectively, than wild-type L. lactis LL27 after 12-h incubation. However, sharp decreases in the yield of nisin were observed at the late phase of fermentation with LAC339 and LL27 in contrast to LAC340 and LAC338 strains for which the high level of nisin could be maintained longer. Obviously, increasing the copy number of the regulation genes together with immunity genes in the nisin producers retarded the loss of nisin in the late phase of the fermentation. PMID- 19137340 TI - Production of nisin Z using Lactococcus lactis IO-1 from hydrolyzed sago starch. AB - A membrane bioreactor for production of nisin Z was constructed using Lactococcus lactis IO-1 in continuous culture using hydrolyzed sago starch as carbon source. A strategy used to enhance the productivity of nisin Z was to maintain the cells in a continuous growth at high cell concentration. This resulted in a volumetric productivity of nisin Z, as 50,000 IU l(-1) h(-1) using a cell concentration of 15 g l(-1), 30( degrees )C, pH 5.5 and a dilution rate of 1.24 h(-1). Adding 10 g l(-1) YE and 2 g l(-1) polypeptone, other inducers were unnecessary to maintain production of nisin. The operating conditions of the reactor removed nisin and lactate, thus minimizing their effects which allowed the maintenance of cells in continuous exponential growth phase mode with high metabolic activity. PMID- 19137339 TI - Microbial analysis of backflowed injection water from a nitrate-treated North Sea oil reservoir. AB - Reservoir souring in offshore oil fields is caused by hydrogen sulphide (H(2)S) produced by sulphate-reducing bacteria (SRB), most often as a consequence of sea water injection. Biocide treatment is commonly used to inhibit SRB, but has now been replaced by nitrate treatment on several North Sea oil fields. At the Statfjord field, injection wells from one nitrate-treated reservoir and one biocide-treated reservoir were reversed (backflowed) and sampled for microbial analysis. The two reservoirs have similar properties and share the same pre nitrate treatment history. A 16S rRNA gene-based community analysis (PCR-DGGE) combined with enrichment culture studies showed that, after 6 months of nitrate injection (0.25 mM NO(3) (-)), heterotrophic and chemolithotrophic nitrate reducing bacteria (NRB) formed major populations in the nitrate-treated reservoir. The NRB community was able to utilize the same substrates as the SRB community. Compared to the biocide-treated reservoir, the microbial community in the nitrate-treated reservoir was more phylogenetically diverse and able to grow on a wider range of substrates. Enrichment culture studies showed that SRB were present in both reservoirs, but the nitrate-treated reservoir had the least diverse SRB community. Isolation and characterisation of one of the dominant populations observed during nitrate treatment (strain STF-07) showed that heterotrophic denitrifying bacteria affiliated to Terasakiella probably contributed significantly to the inhibition of SRB. PMID- 19137341 TI - Carbon utilization profiles of bacteria colonizing the headbox water of two paper machines in a Canadian mill. AB - Forty-one bacterial strains isolated from the headbox water of two machines in a Canadian paper mill were associated with the genera Asticcacaulis, Acidovorax, Bacillus, Exiguobacterium, Hydrogenophaga, Pseudomonas, Pseudoxanthomonas, Staphylococcus, Stenotrophomonas based on the sequence of their 16S rRNA genes. The metabolic profile of these strains were determined using Biolog EcoPlate, and the bacteria were divided into four metabolic groups. Metabolic profiles of the bacterial communities colonizing the headbox water of two paper machines was also determined weekly over a 1 year period. The only compound that was not reduced by the bacterial community was 2-hydroxybenzoic acid. Utilization frequency of the other carbon sources in the Biolog EcoPlate ranged from 3 to 100%. The metabolic profiles of the bacterial community did not vary considerably between the two paper machines. However, the metabolic profile varied among the sampling dates. PMID- 19137343 TI - Neuroscience today: neuronal functional diversity and collective behaviors. Proceedings of an international conference. March 25-27, 2007. Florence, Italy. PMID- 19137342 TI - Characterization of a novel Stenotrophomonas isolate with high keratinase activity and purification of the enzyme. AB - A feather-degrading bacterium was isolated from poultry decomposition feathers in China. The strain, named L1, showed significant feather-degrading activity because it grew and reproduced quickly on basal medium containing 10 g/L of native feather as the source of energy, carbon, and nitrogen. According to the phenotypic characteristics and 16S rRNA profile, the isolate belongs to Stenotrophomonas maltophilia. Keratinase activity of the isolate was determined during cultivation on raw feathers at different temperatures and initial pH. Maximum growth and feather-degrading activity of the bacterium were observed at 40 degrees C and initial pH ranging from 7.5 to 8.0. The crude enzyme was purified by ammonium sulphate precipitation, Sephadex G-100 chromatographic and ceramic hydroxyapatite (CHT) chromatographic. Its molecular mass estimated as 35.2 kDa in SDS-PAGE. The enzyme had an optimum activity at the pH was 7.8 and the temperature was 40 degrees C. The keratinase was wholly inhibited by a serine protease inhibitor, PMSF. Its activity was activated or inhibited by different metal ions. The keratinase activity of enzyme from strain L1 functioned on different keratins, such as feather, hair, wool, horn, and so on. PMID- 19137344 TI - Human factors in GIScience laboratory at the Pennsylvania State University. AB - The human factors in GIScience Laboratory (Human Factors Lab) of The Pennsylvania State University's Department of Geography is located in University Park, PA (USA). University Park and bordering State College, PA are found in the heart of PA between the cities of New York City, NY, Philadelphia, PA, and Pittsburgh, PA. The laboratory is directed by Dr. Alexander Klippel and is part of the GeoVISTA Center. The Human Factors Lab contributes to Penn State Geography's strong tradition as a leader in research on map perception, spatial cognition, and behavior in spatial environments. This report focuses upon basic research topics in spatial cognition, including: (1) perceptual and cognitive factors in map symbolization and design, (2) the creation of cognitively ergonomic route directions for next generation location based services (LBS), (3) You-Are-Here maps and the creation of a sense of place through map-like representations, (4) the conceptualization and representation of dynamic phenomena (i.e., geographic movement pattern), and (5) the relationship between linguistic and non-linguistic conceptualization. PMID- 19137345 TI - Neuroscience Today, Florence 25-27 March 2007: foreword. PMID- 19137347 TI - A simple integrated system for electrophysiologic recordings in animals. AB - This technical note describes a modification to a fundus camera that permits simultaneous recording of pattern electroretinograms (pERGs) and pattern visual evoked potentials (pVEPs). The modification consists of placing an organic light emitting diode (OLED) in the split-viewer pathway of a fundus camera, in a plane conjugate to the subject's pupil. In this way, a focused image of the OLED can be delivered to a precisely known location on the retina. The advantage of using an OLED is that it can achieve high luminance while maintaining high contrast, and with minimal degradation over time. This system is particularly useful for animal studies, especially when precise retinal positioning is required. PMID- 19137346 TI - Neuronal functional diversity and collective behaviors: a scientific case. AB - A major issue in today's neuroscience is how the brain complex and highly flexible organization emerges from its individual components. Robustness of neuronal properties with weak linkages between regulatory processes are suggested to account for the adaptive, tunable, multistable dynamics, the coding schemes and the complexity of neuronal functional (sub)systems. Interneurons and neurotransmitter diversity, resonance phenomena due to properties of the cell or network, time/frequency-dependent activation of dedicated neuronal assemblies, code- and frequency-specific oscillations interact in determining the brain functional setup and operations. Despite the scientific relevance, comprehensive theories are not yet available, but the scenario--however incomplete and incompletely characterized--is promising and warrants further investigation. PMID- 19137348 TI - Visually impaired children: "coming to better terms". AB - For a visually impaired child, the accurate establishment of the diagnosis provides information on the prognosis of his or her participation possibilities, including expectations about the need for care, and provides the basis for informed genetic counseling. To maximize the diagnostic value of electrophysiological testing, we use extensions of the standard ISCEV (International Society for Electrophysiology in Vision) protocols for both the ERG (electroretinogram) and the VEP (visual evoked potential). An overview of 3 years' practice of the Department of Ophthalmology of Bartimeus, presented at ISCEV in Glasgow, showed that, as a result of our electrophysiological assessment, in about 10% of the cases the diagnosis at referral had to be changed from a progressive to a stationary disorder or the reverse. It is obvious that these parameters drastically change the strategy to attain "coming to terms with the disorder". It turns out that for the visually impaired child or his or her parents as well as for the professionals in the rehabilitation institutes, the terminology used to describe a disorder can be unnecessarily alarming rather than comprehensible or even realistic. Terminology needs to be clear and understandable, with a clearcut distinction between the description of visual functions and the name of a disorder. In albinism, the bad connotation of the name of this disorder together with the finding of non-albinos with misrouting and definite albinos without it forces us to reconsider the nomenclature. With congenital stationary night blindness (CSNB), the finding of youngsters who are clearly capable of mobility at night and the fact that the term night blindness refers to a function instead of a disorder forces us even more to reconsider nomenclature. PMID- 19137349 TI - Guidance for catheter ablation of ventricular arrhythmia. PMID- 19137350 TI - Spinal deformity index (SDI) is a good predictor of incident vertebral fractures. AB - The spinal deformity index is a convenient tool to quantify the number and the severity of prevalent vertebral fractures. It is a predictor of the risk of sustaining incident vertebral fracture. This quantification must be taken into account to improve management of patients. INTRODUCTION: Prevalent fractures are strong risk factors for subsequent fractures. METHODS: The study subjects were women from the placebo groups of two studies of strontium ranelate in postmenopausal osteoporosis (N = 723 and 637 patients, respectively). Three lateral radiographs of the spine were obtained at baseline and annually over 3 years, according to standardized procedures. The semiquantitative visual assessment of each vertebra from T4 to L4 was performed by the same reader throughout the study. A spinal deformity index (SDI) was calculated by summing for each patient the grade of each vertebra from T4 to L4. RESULTS: There was a linear relationship between baseline SDI and the 3-year incidence of vertebral fracture (adjusted R(2) = 0.76). The 3-year incidence of vertebral fractures was different among the tertiles of baseline SDI: 17.3 +/- 3.6%, 25.4 +/- 2.6%, and 47.6 +/- 3.1% from the lowest to the highest, respectively. There was no relationship between SDI and non-vertebral fractures incidence. CONCLUSION: SDI is a good predictor of incident vertebral fractures. Patients with highest SDI should receive highest priority to treatment. PMID- 19137351 TI - External hip protectors are effective for the elderly with higher-than-average risk factors for hip fractures. AB - In our cluster randomised controlled trial for efficacy of hip protector with 672 ambulatory elderly women, a hip protector was more effective for prevention of hip fractures in residents with fall history (n = 202; hazard ratio (HR), 0.375; 95%CI, 0.14-0.98; p = 0.05) and body-mass index (BMI) < or = 19.0 (n = 206; HR, 0.37; 95%CI, 0.14-0.95; p = 0.04) by a Cox proportional hazards regression model. INTRODUCTION: Hip fractures result from both osteoporosis and falling. A potentially cost-effective method of preventing hip fractures involves the use of hip protectors but recent studies have revealed the uncertain effectiveness of hip protectors even in institutional settings. METHODS: This study was a cluster randomised controlled trial with nursing homes. We randomly assigned 76 homes with 672 ambulatory but frail elderly women. Several risk factors were assessed at baseline and incorporated into a Cox proportional hazards regression model. UMIN Clinical Trials Registry number is UMIN000000467. Research period was between January 2004 and March 2006. RESULTS: In the intervention group, 19 hip fractures occurred (54.0/1,000 person-years), whereas 39 hip fractures occurred in the control group (78.8/1,000 person-years). Hazard ratio of hip fracture in the intervention group was 0.56 (95%CI, 0.31-1.03; p = 0.06) after adjusting for risk factors. In subgroup analysis, hip protectors were more effective for prevention of hip fractures in residents with fall history (n = 202; HR, 0.375; 95%CI, 0.14-0.98; p = 0.05) and BMI < or = 19.0 (n = 206; HR, 0.37; 95%CI, 0.14 0.95; p = 0.04). Overall compliance with use of hip protectors was 79.7%. CONCLUSION: Risk of hip fracture can be reduced by hip protectors among elderly women with fall history and low BMI. PMID- 19137352 TI - The evaluation of efficacy and tolerability of Hylan G-F 20 in bilateral thumb base osteoarthritis: 6 months follow-up. AB - The aims of this randomized controlled trial were to evaluate the efficacy of intra-articular Hylan G-F 20 on the pain, pinch strength, and functional status in patients with thumb base osteoarthritis (OA). Sixty-six hands of 33 female patients (mean age, 62.6 +/- 6.4 years) with bilateral clinical and radiological thumb base OA were included into this study. Hands of the same patient were divided to Hylan G-F 20 and saline-injection groups. VAS pain 100 mm, pinch strength (key, tripod and pulp to pulp pinch) and functionality (Dreiser functional index) were assessed at baseline, at sixth and 24th weeks. Statistically significant improvements were detected in function (p = 0.001), VAS pain (p = 0.002), and pinch strength (p = 0.004) at the 24th week in the Hylan G F 20 group. However, only VAS pain scores decreased temporarily in control hands at the 6th week (p = 0.02). Although short-term placebo analgesic effect could not be ignored, intra-articular hylan was effective on pain, pinch strength, and function at the 24th week. PMID- 19137353 TI - Comparison of therapeutic effects of sodium hyaluronate and corticosteroid injections on trapeziometacarpal joint osteoarthritis. AB - This was a randomized, open-label, evaluator-blinded clinical study including 40 women with stage II or III trapeziometacarpal joint osteoarthritis. The steroid group (n = 20) received one injection of 20 mg triamcinolone acetonide once and the hyaluronate group (n = 20) received three injections of 5 mg sodium hyaluronate at 1-week intervals. The pain level was assessed using a visual analog scale and grip and pinch strengths were measured using a hand grip dynomemeter and pinch gauge. The Duruoz Hand Index was used to evaluate hand function. Pain level decreased significantly over 12 months for the steroid group and over 6 months for the sodium hyaluronate group. Pinch strength did not improve in either group, but grip strength improved significantly in both groups. Hand function improved in both groups but it was only significant in the steroid group. Our findings showed that both intra-articular injection of steroid and sodium hyalurunate are effective in trapeziometacarpal joint osteoarthritis. However the steroid injection was found to be superior to sodium hyaluronate injection in reducing pain and improving hand function. PMID- 19137354 TI - Translation and validation of the Persian version of the Arthritis Impact Measurement Scales 2-Short Form (AIMS2-SF) in patients with rheumatoid arthritis. AB - Cultural adaptation and validation of the Persian version of the Arthritis Measurement Scales 2-Short Form (AIMS2-SF). The translation and cultural adaptation of the original questionnaire was carried out in accordance with published guidelines. Three hundred and fifty consecutive Persian-speaking patients with rheumatoid arthritis (RA) were asked to complete the AIMS2-SF, the Short Form Health Survey (SF-36), and four visual analog scales (VAS) for pain, joint stiffness, and patient's and physician's global assessment to test convergent validity. In addition, 90 randomly selected patients were asked to complete the questionnaire 48 h later for the second time. Moderate to high correlation were found between the AIMS2-SF and subscales of the SF-36 and VAS for pain, morning stiffness, and patient's and physician's global assessment. Cronbach's alpha coefficient for the Persian AIMS2-SF scales ranged from 0.74 to 0.89. The Persian AIMS2-SF scales showed excellent test-retest reliability with Intraclass Correlation Coefficient ranging from 0.83-0.93 (p < 0.01). The results of the present study showed that the Persian AIMS2-SF has reasonably good convergent validity, internal consistency, and test-retest reliability in patients with RA. It can now be applied in clinical settings and future outcome studies in Iran. PMID- 19137355 TI - Expression of multidrug resistance-1 protein correlates with disease activity rather than the refractoriness to methotrexate therapy in rheumatoid arthritis. AB - Disease-modifying antirheumatic drugs (DMARDs) improve the disability and slow the progression of the joint damage in rheumatoid arthritis (RA). However, a large proportion of patients experience inefficacy by the end of 2 years. This loss of efficacy may be due to expression of multidrug resistance (MDR) proteins on lymphocytes. The objective is to study the expression of MDR protein on the peripheral blood lymphocytes in patients with RA and correlate it with the disease status and response to treatment. Twenty-eight patients were enrolled. Expression of MDR-1 by flow cytometry was carried out on lymphocytes at baseline and after 4 months of therapy. This expression was correlated with disease activity scores (DAS 28). There were 25 females with mean age of 48.13 years and median disease duration of 48 months. Eighteen patients were DMARD naive and ten were refractory to DMARD (methotrexate). The percentage of cells expressing MDR-1 in the DMARD-naive (p<0.O5) and DMARD-refractory (p<0.05) groups were significantly higher than the healthy controls at the baseline. The relative fluorescence intensity was significantly higher in the DMARD-refractory group (p<0.05) as compared to the DMARD-naive group. After 4 months of therapy, there was significant improvement in the D value (p<0.01) in the DMARD-naive group (treated with methotrexate only) and DMARD-refractory group (p<0.05). A significant correlation (r=0.563) between the DAS 28 scores and the D value (p=0.003) was observed. Expression of MDR-1 in RA correlated with disease activity status and improved with DMARD therapy. It is not related to the refractoriness to therapy with methotrexate. PMID- 19137363 TI - A prospective randomized trial comparing traditional and fast-track patient care in elective open infrarenal aneurysm repair. AB - BACKGROUND: Fast-track recovery programs have led to reduced patient morbidity and mortality after major surgery. In terms of elective open infrarenal aneurysm repair, no evidence is available about such programs. To address this issue, we have conducted a randomized prospective pilot study. METHODS: The study involved prospective randomization of 101 patients with the indication for elective open aneurysm repair in a traditional and a fast-track treatment arm. The basic fast track elements were no bowel preparation, reduced preoperative fasting, patient controlled epidural analgesia (PCEA), enhanced postoperative feeding, and postoperative mobilization. Morbidity and mortality, need for postoperative mechanical ventilation, length of stay (LOS) in the intensive care unit (ICU) and total length of postoperative hospital stay were analyzed in terms of an intention to treat. RESULTS: Demographic data for the two groups were similar. In the fast-track group the need for postoperative ventilation was significantly lower (6.1% versus 32%; p = 0.002), the median LOS on ICU did not significantly differ (20 h versus 32 h; p = 0.183), full enteral feeding was achieved significantly earlier (5 versus 7 days; p < 0.0001), and the rate of postoperative medical complications-gastrointestinal, cardiac, pulmonary, renal, and infective-was significantly lower (16% versus 36%; p = 0.039). The postoperative hospital stay was significantly shorter in the fast-track group (10 days versus 11 days; p = 0.016); the mortality rate in both groups was 0%. CONCLUSIONS: An optimized patient care program in open infrarenal aortic aneurysm repair shows favorable results concerning need for postoperative assisted mechanical ventilation, time to full enteral feeding, and incidence of medical complications. Further ranomized multicentric trials are necessary to justify broad implementation (clinical trials. gov identifier NCT 00615888). PMID- 19137362 TI - Microarray analysis of somatostatin receptor 5-regulated gene expression profiles in murine pancreas. AB - BACKGROUND: We previously demonstrated that somatostatin receptor type 5 (SSTR5) gene ablation results in alterations in insulin secretion and glucose metabolism, accompanied by morphologic alterations in the islets of Langerhans. The underlying mechanism(s) by which SSTR5 exerts its cellular functions remain(s) unknown. We hypothesized that SSTR5 mediates the inhibitory effect of somatostatin (SST) on insulin secretion and islet proliferation by regulating a specific set of pancreatic genes. METHODS: To identify SSTR5-regulated pancreatic genes, gene expression microarray analysis was performed on the whole pancreas of 1- and 3-month-old wild-type (WT) and SSTR5 knockout (SSTR5-/-) male mice. Real time RT-PCR and immunofluorescence were performed to validate selected differentially expressed genes. RESULTS: A set of 143 probes were identified to be differentially expressed in the pancreas of 1-month-old SSTR5-/- mice, 72 of which were downregulated and 71 upregulated. At 3 months of age, SSTR5 gene ablation resulted in downregulation of a set of 30 probes and upregulation of a set of 37 probes. Among these differentially expressed genes, there were 15 and 5 genes that were upregulated and downregulated, respectively, in mice at both 1 and 3 months of age. Three genes, PAP/INGAP, ANG, and TDE1, were selected to be validated by real-time RT-PCR and immunofluorescence. CONCLUSIONS: A specific set of genes linked to a wide range of cellular functions such as islet proliferation, apoptosis, angiogenesis, and tumorigenesis were either upregulated or downregulated in SSTR5-deficient male mice compared with their expression in wild-type mice. Therefore, these genes are potential SSTR5-regulated genes during normal pancreatic development and functional maintenance. PMID- 19137364 TI - Chest computed tomography screening in colorectal cancer patients. PMID- 19137365 TI - Safety of laparoscopic appendectomy during pregnancy. AB - BACKGROUND: Use of laparoscopic appendectomy (LA) remains controversial during pregnancy because data regarding procedure safety are limited. The outcome of LA in pregnant women was evaluated and compared to results of open surgery. METHODS: Between January 1997 and December 2007, 42 pregnant women (mean age 24 years [range: 19-40 years]; range of gestation: 5-25 weeks) underwent appendectomy for suspected acute appendicitis: 23 laparoscopic (LA) and 19 open appendectomies (OA). Retrospective review of medical charts included preoperative information, surgery results, and outcome of the pregnancy. RESULTS: There was no difference between groups in surgery delay following arrival at the hospital. All procedures, except one case of Meckel's diverticulitis, were completed laparoscopically without need for conversion. Acute appendicitis was found in 19 cases and Meckel's diverticulitis in one case during LA (87%) and in 18 cases (95%) during OA. Complicated appendicitis was found in 7 (30%) pregnant women in the LA group and 1 (5%) in the OA group. Five women with normal preoperative abdominal ultrasonography had acute appendicitis (2 in the OA group and 3 in the LA group). The laparoscopic procedure was performed more often by senior surgeons (70% of cases), and OA was more commonly done by residents (47% of cases). There were no intraoperative or postoperative complications recorded. The length of postoperative hospital stay was slightly prolonged after LA-2.4 days versus 1.4 day after OA. There was one fetal loss in each group, 1 and 2 months after the operation. CONCLUSIONS: Laparoscopic appendectomy is safe and effective during pregnancy and is associated with good maternal and fetal outcome. PMID- 19137366 TI - Differences in the status of involved lymph nodes and the direction of metastatic lymphatic flow from tumors into involved nodes between submucosal and T2-4 thoracic squamous cell esophageal cancers. PMID- 19137367 TI - Surgical approach to extensive hidradenitis suppurativa in the perineal/peri-anal and gluteal regions. PMID- 19137368 TI - Developing a tissue resource to characterize the genome of pancreatic cancer. AB - With recent advances in DNA sequencing technology, medicine is entering an era in which a personalized genomic approach to diagnosis and treatment of disease is feasible. However, discovering the role of altered DNA sequences in various disease states will be a challenging task. The genomic approach offers great promise for diseases, such as pancreatic cancer, in which the effect of current diagnostic and treatment modalities is disappointing. To facilitate the characterization of the genome of pancreatic cancer, high-quality and well annotated tissue repositories are needed. This article summarizes the basic principles that guide the creation of such a repository, including sample processing and preservation techniques, sample size and composition, and collection of clinical data elements. PMID- 19137369 TI - Embolization of the gastroduodenal artery before selective internal radiotherapy: a prospectively randomized trial comparing platinum-fibered microcoils with the Amplatzer Vascular Plug II. AB - The Amplatzer Vascular Plug II (AVP II) is a novel device for transcatheter vessel occlusion, for which only limited comparative data exist. Embolotherapy of the gastroduodenal artery (GDA) is essential before internal radiotherapy (SIRT) in order to prevent radiation-induced peptic ulcerations due to migration of yttrium-90 microspheres. The purpose of this study was to compare the vascular anatomical limitations, procedure time, effectiveness, and safety of embolization of the GDA with coils versus the AVP II. Fifty patients stratified for SIRT were prospectively randomized for embolization of the GDA with either coils or the AVP II. The angle between the aorta and the celiac trunk, diameter of the GDA, fluoroscopy time and total time for embolization, number of embolization devices, complications, and durability of vessel occlusion at follow-up angiography for SIRT were recorded. A t-test was used for statistical analysis. Embolizations with either coils or the AVP II were technically feasible in all but two patients scheduled for embolization of the GDA with the AVP II. In both cases the plug could not be positioned due to the small celiac trunk outlet angles of 17 degrees and 21 degrees. The mean diameter of the GDA was 3.7 mm (range, 2.2-4.8 mm) for both groups. The procedures differed significantly in fluoroscopy time (7.8 min for coils vs. 2.6 min for the AVP II; P < 0.001) and embolization time (23.1 min for coils vs. 8.8 min for the AVP II; P < 0.001). A mean of 6.0 +/- 3.2 coils were used for GDA embolization, while no more than one AVP II was needed for successful vessel occlusion (P < 0.001). One coil migration occurred during coil embolization, whereas no procedural complication was encountered with the use of the AVP II. Vessel reperfusion was noted in only one patient, in whom coil embolization was performed. In conclusion, embolization of the GDA with the AVP II is safe, easy, rapid, and highly effective; only an extremely sharp-angled celiac trunk outlet represented an anatomical limitation for device deployment. PMID- 19137370 TI - Abdominal chyloma: CT findings and percutaneous drainage. PMID- 19137371 TI - Plant growth-promoting and rhizosphere-competent Acinetobacter rhizosphaerae strain BIHB 723 from the cold deserts of the Himalayas. AB - A phosphate-solubilizing bacterial strain BIHB 723 isolated from the rhizosphere of Hippophae rhamnoides was identified as Acinetobacter rhizosphaerae on the basis of phenotypic characteristics, carbon source utilization pattern, fatty acid methyl esters analysis, and 16S rRNA gene sequence. The strain exhibited the plant growth-promoting attributes of inorganic and organic phosphate solubilization, auxin production, 1-aminocyclopropane-1-carboxylate deaminase activity, ammonia generation, and siderophore production. A significant increase in the growth of pea, chickpea, maize, and barley was recorded for inoculations under controlled conditions. Field testing with the pea also showed a significant increment in plant growth and yield. The rifampicin mutant of the bacterial strain effectively colonized the pea rhizosphere without adversely affecting the resident microbial populations. PMID- 19137373 TI - The effect of wireless LAN-based PACS device for portable imaging modalities. AB - The aim of this study was to develop wireless Picture Archiving and Communication System (PACS) device and to analyze its effect on image transfer from portable imaging modalities to the main PACS server. Using a laptop computer equipped with wireless local area network (LAN), the authors developed a wireless PACS device with DICOM modality worklist and DICOM storage server modules. This laptop computer could be easily fixed to portable imaging modalities such as ultrasound machines. From May to August 2007, 112 portable examinations were evaluated. Of these, 62 were done with wireless LAN-based PACS device, and 50 were done without wireless PACS device. To evaluate the impact of the wireless LAN-based PACS device on productivity and workflow, we analyzed the mean time delay and standard deviations (SD) both in cases where wireless LAN-based PACS device was used and in cases where it was not used. Statistical analysis was performed using a t test. The mean time interval from image acquisition to storage in the main PACS when the wireless LAN-based PACS device was used was 342.4 s (5 min and 42.4 s, SD = 509.2 s). When the wireless PACS was not used, the mean time interval was 2,305.5 s (38 min and 25.5 s, SD = 1,371.8 s). The mean time interval was statistically different between the two groups (t test, p < 0.001). The wireless LAN-based PACS device could help in reducing the storage intervals of images obtained by portable machines and in promoting effective and rapid treatment of patients who have undergone portable imaging examinations. PMID- 19137372 TI - Genetic factors for resistance to diet-induced obesity and associated metabolic traits on mouse chromosome 17. AB - Obesity is associated with increased susceptibility to dyslipidemia, insulin resistance, and hypertension, a combination of traits that comprise the traditional definition of the metabolic syndrome. Recent evidence suggests that obesity is also associated with the development of nonalcoholic fatty liver disease (NAFLD). Despite the high prevalence of obesity and its related conditions, their etiologies and pathophysiology remains unknown. Both genetic and environmental factors contribute to the development of obesity and NAFLD. Previous genetic analysis of high-fat, diet-induced obesity in C57BL/6J (B6) and A/J male mice using a panel of B6-Chr(A/J)/NaJ chromosome substitution strains (CSSs) demonstrated that 17 CSSs conferred resistance to high-fat, diet-induced obesity. One of these CSS strains, CSS-17, which is homosomic for A/J-derived chromosome 17, was analyzed further and found to be resistant to diet-induced steatosis. In the current study we generated seven congenic strains derived from CCS-17, fed them either a high-fat, simple-carbohydrate (HFSC) or low-fat, simple carbohydrate (LFSC) diet for 16 weeks and then analyzed body weight and related traits. From this study we identified several quantitative trait loci (QTLs). On a HFSC diet, Obrq13 protects against diet-induced obesity, steatosis, and elevated fasting insulin and glucose levels. On the LFSC diet, Obrq13 confers lower hepatic triglycerides, suggesting that this QTL regulates liver triglycerides regardless of diet. Obrq15 protects against diet-induced obesity and steatosis on the HFSC diet, and Obrq14 confers increased final body weight and results in steatosis and insulin resistance on the HFSC diet. In addition, on the LFSC diet, Obrq 16 confers decreased hepatic triglycerides and Obrq17 confers lower plasma triglycerides on the LFSC diet. These congenic strains provide mouse models to identify genes and metabolic pathways that are involved in the development of NAFLD and aspects of diet-induced metabolic syndrome. PMID- 19137374 TI - Dual-isotope acquisition for CT-SPECT registration of infection studies. AB - The registration of CT and NM images can enhance patient diagnosis since it allows for the fusion of anatomical and functional information as well as attenuation correction of NM images. However, irrespective of the methods used, registration accuracy depends heavily on the characteristics of the input images and the degree of similarity between them. This poses a challenge for registering CT and NM images as they may have very different characteristics. To address the particular problem of CT and In-111 SPECT registration, we propose to perform a dual-isotope study which involves an additional injection of Tc-99m MDP to generate two inherently registered images: In-111 SPECT and Tc-99m SPECT. As skeletal structures are visible in both CT and Tc-99m SPECT, performing registration of these images may be much more effective. The very same spatial transformation derived can be immediately applied to complete the registration of CT and the corresponding In-111 SPECT. Accordingly, we hypothesize that the registration of CT and Tc-99m SPECT can be more accurately performed than the registration of CT and In-111 SPECT and seek to compare the accuracies between the aforementioned registrations. In this paper, we have collected three clinical datasets, with the ground-truth transformations known, and tested the proposed approach by using a mutual information-based algorithm to solve for the rigid/non rigid misalignments introduced to them. Based on the results of our experiments, we conclude that registration using Tc-99m SPECT can achieve 100% success rate, and is thus much more superior to the registration using In-111 SPECT, which at best, achieves only 38% success rate. Clearly, the introduction of a dual-isotope acquisition can substantially improve the registration of SPECT and CT images. PMID- 19137375 TI - The prognostic impact of isolated tumor cells in lymph nodes of T2N0 gastric cancer: comparison of American and Japanese gastric cancer patients. AB - BACKGROUND: The clinical significance of immunohistochemically detected isolated tumor cells (ITC) in lymph nodes of gastric cancer patients is controversial. This study examined the prognostic impact of ITC on patients with early-stage gastric cancer in two large volume centers in the United States and Japan. METHODS: Fifty-seven patients with T2N0M0 gastric carcinoma who underwent gastric resection between January 1987 and January 1997 at Memorial Sloan-Kettering Cancer Center (MSKCC) in New York and 107 patients resected at National Cancer Center Hospital (NCCH) in Tokyo between January 1984 and December 1990 were studied. The sections were newly prepared from each lymph node for immunohistochemical staining for cytokeratin. Lymph nodes and original specimens from MSKCC were examined by pathologists in NCCH. The prognostic significance of the presence of ITC in lymph nodes was investigated in patients of both institutions. RESULTS: ITC were identified in 30 of 57 patients (52.6%) at MSKCC and in 38 of 107 patients (35.5%) at NCCH. In both institutions, there was no significant difference in the prognosis of the studied patients with or without ITC (P= .22, .86 respectively). CONCLUSIONS: The presence of ITC detected by immunohistochemistry in the regional lymph nodes did not affect the prognosis of American and Japanese patients with T2N0M0 gastric carcinoma who underwent gastrectomy with D2 lymph node dissection. PMID- 19137376 TI - Automated microscopy to evaluate surgical specimens via touch prep in breast cancer. AB - BACKGROUND: Breast conservation therapy is the standard treatment for breast cancer; however, 20-50% of operations have a positive margin leading to secondary procedures. The standard of care to evaluate surgical margins is based on permanent section. Imprint cytology (touch prep) has been used to evaluate surgical samples, but conventional techniques require an experienced cytopathologist for correct interpretation. An automated image screening process has been developed to discern cancer cells from normal epithelial cells. This technique is based on cellularity of the imprint specimen and does not require expertise in cytopathology. METHODS: A rapid immunofluorescent staining technique coupled with automated microscopy was used to classify specimens as cancer vs. noncancer based on the density of epithelial cells captured on touch prep of tumor cross-sections. The results of the automated analysis vs. a manual screen of ten 20x fields were compared to the pathology interpretation on permanent section. RESULTS: A total of 34 consecutive cases were analyzed: 10 normal cases, and 24 cancer cases. The cross-section specimens for invasive cancer were correctly classified in at least 65% of the cases by using manual microscopy and at least 83% by using automated microscopy. The manual and automated microscopy correlated well for measurements of epithelial cell density (R(2)=0.64); however, the automated microscopy was more accurate. CONCLUSIONS: This preliminary study using an automated system for intraoperative interpretation does not require a cytopathologist and shows that rapid, low-resolution imaging can correctly identify cancer cells for invasive carcinoma in surgical specimens. Therefore, automated determination of cellularity in touch prep is a promising technique for future margin interpretation of breast conservation therapy. PMID- 19137378 TI - Limitations of conduits in peripheral nerve repairs. AB - Nerve conduits have emerged as alternatives to autologous nerve grafts, but their use in large-diameter nerve deficits remains untested. We report four patients who underwent repair of large-diameter nerves using absorbable nerve conduits and discuss the failed clinical outcomes. The reported cases demonstrate the importance of evaluating the length, diameter, and function of nerves undergoing conduit repair. In large-diameter nerves, the use of conduits should be carefully considered. PMID- 19137379 TI - Patient use of secure electronic messaging within a shared medical record: a cross-sectional study. AB - BACKGROUND: Most patients would like to be able to exchange electronic messages with personal physicians. Few patients and providers are exchanging electronic communications. OBJECTIVE: To evaluate patient characteristics associated with the use of secure electronic messaging between patients and health care providers. DESIGN, SETTING, AND PATIENTS: Cross-sectional cohort study of enrollees over 18 years of age who were enrolled in an integrated delivery system in 2005. MEASUREMENTS AND MAIN RESULTS: Among eligible enrollees, 14% (25,075) exchanged one or more secure messages with a primary or specialty care provider between January 1, 2004 and March 31, 2005. Higher secure messaging use by enrollees was associated with female gender (OR, 1.15; 95% CI, 1.10-1.19), greater overall morbidity (OR, 5.64; 95% CI, 5.07-6.28, comparing high or very high to very low overall morbidity), and the primary care provider's use of secure messaging with other patients (OR, 1.94; 95% CI, 1.67-2.26, comparing 20 50% vs. 420 min) and advanced age (>70 years) were the only prognostic variables, which significantly diminished survival on multivariate analysis. CONCLUSION: Resection of the superior mesenteric or portal vein to achieve macroscopic tumor clearance can be performed safely with acceptable operative morbidity and mortality. However, improved local clearance in these patients cannot achieve a favorable long-term survival for all patients because distant metastases or local recurrence is frequent. PMID- 19137381 TI - Changes in lipid profile and insulin resistance in obese patients after Scopinaro biliopancreatic diversion. AB - BACKGROUND: Obesity is associated with many cardiovascular risk factors. The aim of this study is to evaluate the effect of bariatric surgery (BS) in lipid profile and insulin resistance in obese patients. MATERIAL AND METHODS: We studied changes in lipid parameters glucose, insulin, and insulin resistance (IR) before biliopancreatic diversion and 3, 6, 12, 18, and 24 months after surgery, in 115 obese patients divided in two groups: diabetics (DM) and nondiabetics (non DM). RESULTS: In both groups, all parameters significantly decreased at 3 months; however, high-density lipoprotein cholesterol levels (HDL-c) and total cholesterol (TC) to HDL-c ratio decreased up to 6 months. At baseline, in DM, we found a negative correlation between body mass index (BMI) and TC to HDL-c ratio and glucose with HDL-c and homeostasis model assessment (HOMA) with BMI and a direct correlation between glucose with TC to HDL-c ratio and glucose with triglycerides. In non-DM, no correlations were found. Comparing both groups for each parameter, we found significant differences at basal levels for glucose, HOMA, triglycerides (TG), and TG to HDL-c ratio. Changes between basal levels and 1 year after surgery were significantly different in glucose and HOMA. CONCLUSION: BS offers excellent results in weight reduction and significantly improves IR, diabetes, and the lipid profile, decreasing global cardiovascular risk. PMID- 19137387 TI - Genetic variation of Picea jezoensis populations in South Korea revealed by chloroplast, mitochondrial and nuclear DNA markers. AB - Genetic variation associated with Picea jezoensis populations of South Korea was investigated using chloroplast (cp), mitochondrial (mt) and nuclear DNA markers. In South Korea, P. jezoensis is distributed across a very restricted area, being found on the summits of three mountains: Mts. Jiri, Dokyu and Gyebang. Examination of five region restriction enzyme combinations for mtDNA and four for cpDNA revealed haplotypes endemic to South Korea. The Gyebang population, the most northerly and most isolated, was genetically distinct from the other populations. Nuclear microsatellite markers indicated, overall, a low level of genetic diversity (H (e) = 0.406) in South Korea; this could be attributed to genetic drift and/or founder effects associated with historical events. The Wilcoxon sign-rank test did not indicate a recent bottleneck in any of the populations irrespective of the model considered (infinite allele model, two phased model of mutation, and stepwise mutation model). Microsatellite markers also demonstrated that the Gyebang population was distinct from the others. The results of this study could be used as the basis for conservation guidelines for the management of this species in South Korea. PMID- 19137388 TI - The 60th anniversary of Japanese primatology. PMID- 19137382 TI - PTPRR protein tyrosine phosphatase isoforms and locomotion of vesicles and mice. AB - Protein tyrosine phosphatases (PTPs) are central players in many different cellular processes and their aberrant activity is associated with multiple human pathologies. In this review, we present current knowledge on the PTPRR subfamily of classical PTPs that is expressed in neuronal cells and comprises receptor-type (PTPBR7, PTP-SL) as well as cytosolic (PTPPBSgamma-37, PTPPBSgamma-42) isoforms. The two receptor-type isoforms PTPBR7 and PTP-SL both localize in late endosomes and the Golgi area. PTPBR7, however, is additionally localized at the cell surface and on early endosomes. During cerebellar maturation, PTPBR7 expression in developing Purkinje cells ceases and is replaced by PTP-SL expression in the mature Purkinje cells. All PTPRR isoforms contain a kinase interacting motif that makes them mitogen-activated protein kinase phosphatases. The distinct subcellular localization of the different PTPRR isoforms may reflect differential roles in growth-factor-induced MAPK-mediated retrograde signaling cascades. Studies in PTPRR-deficient mice established that PTPRR isoforms are physiological regulators of MAPK phosphorylation levels. Surprisingly, PTPRR-deficient mice display defects in motor coordination and balancing skills, while cerebellar morphological abnormalities, which are often encountered in ataxic mouse models, are absent. This is reminiscent of the phenotype observed in a handful of mouse mutants that have alterations in cerebellar calcium ion homeostasis. Elucidation of the molecular mechanisms by which PTPRR deficiency imposes impairment of cerebellar neurons and motor coordination may provide candidate molecules for hereditary cerebellar ataxias that still await identification of the corresponding disease genes. PMID- 19137389 TI - Chimpanzee social intelligence: selfishness, altruism, and the mother-infant bond. AB - To better understand the human mind from an evolutionary perspective, a great deal of research has focused on the closest living relative of humans, the chimpanzee, using various approaches, including studies of social intelligence. Here, I review recent research related to several aspects of social intelligence, including deception, understanding of perception and intention, social learning, trading, cooperation, and regard for others. Many studies have demonstrated that chimpanzees are proficient in using their social intelligence for selfish motives to benefit from their interactions with others. In contrast, it is not yet clear whether chimpanzees engage in prosocial behaviors that benefit others; however, chimpanzee mother-infant interactions indicate the possibility of such behaviors. Therefore, I propose that chimpanzees possess rudimentary traits of human mental competence not only in terms of theory of mind in a broader sense but also in terms of prosociality involving regard for others. Mother-infant interactions appear to be particularly important to understanding the manifestation of social intelligence from an evolutionary perspective. PMID- 19137390 TI - Emergence, propagation or disappearance of novel behavioral patterns in the habituated chimpanzees of Mahale: a review. AB - Each local population of chimpanzees shows cultural variation, but little is known about how behavioral variations first emerge, and how often variants spread to other individuals and then become fixed as a local culture in chimpanzee society. Although field studies of chimpanzees are still too short to answer these questions definitively, it may stimulate further study in various sites to summarize the developments observed over the past 40 years at Mahale, Tanzania. Innovative patterns were operationally defined as new behavioral patterns performed by M group chimpanzees from 1981 onwards. Innovations included patterns of feeding (n = 8), human-directed behavior (n = 3), hygiene behavior (n = 4), maternal carrying of infants (n = 2), courtship (n = 2), play (n = 6), intimidation displays (n = 3), and quasi-grooming (n = 4). Although most patterns were repeated later by other individuals, six patterns were never seen performed by another individual, and eight patterns were performed by one or a few individuals but social transmission was unlikely. Thus, innovation was not rare, but emergence of fashion or establishment of traditions seems to occur rarely in chimpanzee society. PMID- 19137391 TI - [Editorial to the main topic: forefoot surgery]. PMID- 19137392 TI - [The proximal open-wedge osteotomy with interlocking plate for correction of splayfoot deformities with hallux valgus]. AB - OBJECTIVE: Correction of splayfoot deformity through reduction of the increased first intermetatarsal angle by a proximal open-wedge osteotomy. INDICATIONS: Splayfoot deformity with a first intermetatarsal angle > 14 degrees and hallux valgus deformity in younger patients. Splayfoot deformities with a short first metatarsal. CONTRAINDICATIONS: Degenerative changes in the first metatarsophalangeal joint. Contractures of the first metatarsophalangeal joint. Relative: overlength of the first metatarsal. Relative: lateral tilt of the articular cartilage surface of the first metatarsal head. SURGICAL TECHNIQUE: Proximal metatarsal osteotomy approximately 10 mm distal of the first tarsometatarsal joint with preservation of the lateral cortex. Careful manual opening of the osteotomy to avoid a fracture of the lateral cortex. Fixation of the osteotomy with an interlocking plate. Filling of the defect with cancellous bone. Distal soft-tissue procedure. POSTOPERATIVE MANAGEMENT: Postoperative hallux shoe for 6 weeks. Partial weight bearing (20 kg) for 2 weeks, afterwards full weight bearing. Mobilization of the first metatarsophalangeal joint. Full weight bearing in comfortable shoes after appropriate bony healing has occurred radiologically. Sports with a high impact on the foot is allowed after 12 weeks. Orthotics are prescribed, if some pain remains or associated pathologies require external support. RESULTS: In a consecutive series of 35 patients, the first intermetatarsal angle could be reduced to normal values (6.8 degrees , standard deviation 1.3 degrees ). The open-wedge osteotomy resulted in a slight lengthening of the first metatarsal (1-2 mm). In one patient local wound healing problems had to be treated with oral antibiotics. Operative revision was not necessary in any case. Implant failure or nonunion was not observed. One plate was removed because of local subcutaneous irritation. PMID- 19137393 TI - [The retrocapital osteotomy ("chevron") for correction of splayfoot with hallux valgus]. AB - OBJECTIVE: Surgical treatment of hallux valgus deformity with a distal osteotomy of the first metatarsal to address an increased intermetatarsal angle (IMA) I-II. This procedure is combined with a soft-tissue procedure at the first metatarsophalangeal joint: realignment of the first ray, lateral displacement of the first metatarsal head above the sesamoids, rebalancing of the soft tissues at the metatarsophalangeal joint. INDICATIONS: Pain and soft-tissue inflammation at the bunion, impaired function of the metatarsophalangeal joint, and lateral deviation of the hallux. IMA I-II 10 degrees. CONTRAINDICATIONS: Symptomatic osteoarthritis of the first metatarsophalangeal joint, assessed clinically or radiographically. Acute inflammation of the forefoot, osteoporosis of the first metatarsal. Vascular disturbance. Cosmetic indication only. Relative: hypermobility of the first ray, valgus malalignment of the hindfoot, previous retrocapital osteotomy. SURGICAL TECHNIQUE: Lateral soft tissue release. Resection of the medial pseudoexostosis. V-shaped osteotomy of the distal metatarsal I. Exostosectomy. Lateral displacement of the first metatarsal head. Screw fixation. Realignment of the metatarsophalangeal joint by tightening of the medial soft tissues. POSTOPERATIVE MANAGEMENT: Postoperative shoe with full weight bearing. Active exercises of the foot and hallux. Physiotherapy. Prophylaxis of deep vein thrombosis depending on the degree of mobility. Radiographic control after 6 weeks. Bandage or orthosis to maintain toe alignment. RESULTS: IMA I-II was reduced from 13.6 degrees preoperatively to 6.6 degrees postoperatively. HVA decreased from 29.8 degrees to 8.2 degrees postoperatively. PMID- 19137394 TI - [The Akin procedure as closing wedge osteotomy for the correction of a hallux valgus interphalangeus deformity]. AB - OBJECTIVE: Realignment of the great toe in the case of a hallux valgus interphalangeus by means of a medially based closing wedge osteotomy. INDICATIONS: Hallux valgus interphalangeus deformity, characterized by an enlarged distal articular surface angle (> 10 degrees). Correction of a hallux valgus interphalangeus deformity as an additional procedure in the case of hallux valgus surgery. CONTRAINDICATIONS: Incongruent first metatarsophalangeal joint with lateral subluxation of the proximal phalanx. Isolated procedure to correct hallux valgus deformity. Lack of patient compliance. Neurovascular disturbance of the forefoot. SURGICAL TECHNIQUE: Medially based closing wedge osteotomy of the proximal phalanx to reduce the distal articular surface angle. Fixation with a lag screw, cannulated Herbert screw, memory cramp, threaded Kirschner wire, or interosseous suture. POSTOPERATIVE MANAGEMENT: Wound dressing to assure the position of the great toe. Radiographic documentation of the forefoot in two planes. Strict elevation of the operated foot to prevent postoperative swelling. Mobilization of the patient with a forefoot relief orthosis, until consolidation of the osteotomy is verified radiologically (4-5 weeks). Low-molecular-weight heparin for at least 1 week. Hallux valgus bandage or functional taping for 6 weeks postoperatively in patients with additional metatarsal osteotomy. RESULTS: Clinical and radiologic follow-up based on 32 patients showed good results. The postoperative Hallux Score of the American Orthopaedic Foot and Ankle Society improved to 89 points. PMID- 19137395 TI - [Cheilectomy and Kessel-Bonney procedure for treatment of initial hallux rigidus]. AB - OBJECTIVE: Joint-preserving procedure for initial osteoarthritis of the first metatarsophalangeal joint for improvement of restricted joint motion and achievement of a harmonic gait. INDICATIONS: Hallux rigidus stage I and II according to Regnauld's classification. CONTRAINDICATIONS: Hallux rigidus Regnauld stage III. General medical contraindications to surgical interventions and anesthesiological procedures. SURGICAL TECHNIQUE: Operation in regional anesthesia (foot block). Tourniquet. Longitudinal skin incision over the dorsal aspect of the first metatarsophalangeal joint. Incision of the joint capsule with protection of the extensor hallucis longus tendon and the dorsal neurovascular bundle. Cheilectomy: removal of osteophytes at the metatarsal head and the base of the proximal phalanx. Resection of the dorsal third of the metatarsal head with an oscillating saw in plantar flexion of the proximal phalanx. Kessel-Bonney procedure: dissection of the proximal phalanx. Incomplete dorsal osteotomy with an oscillating saw at the metaphysis of the proximal phalanx and removal of a dorsal wedge with a base of 2-3 mm. Osteosynthesis with mini-plate or transosseous suture. POSTOPERATIVE MANAGEMENT: Postoperative elevation of the operated foot. Analgesia with nonsteroidal anti-inflammatory drugs. Postoperative shoe for 3-4 weeks. Immediate weight bearing. Mobilization of the metatarsophalangeal joint with an elastic bandage. Taping in extension and elastic forefoot dressing for 3 weeks postoperatively. Clinical and radiologic controls after 6 and 12 weeks. RESULTS: 53 operations on 45 patients were performed. 39 patients (86%; 28 female, eleven male, mean age 43.6 years) were followed up. After a period of 26 months (range: 10-51 months), 32 of 39 patients (82%) were satisfied or very satisfied. The median preoperative range of motion was 12.3 degrees for dorsal extension and 17.1 degrees for plantar flexion. Function had increased to a dorsiflexion of 34.2 degrees and a plantar flexion of 32.8 degrees. All patients returned to a normal walking ability after a mean period of 3.7 weeks. Due to delayed wound healing, one revision was necessary. According to Kitaoka's Forefoot Score, the mean preoperative value of 44.3 (standard deviation [SD]: +/- 16) increased postoperatively to a mean value of 78.9 (SD: +/- 12). PMID- 19137396 TI - [The Valenti resection arthroplasty in the treatment of advanced hallux rigidus]. AB - OBJECTIVE: With this joint-preserving procedure impinging and damaged parts of the first metatarsal head and the proximal phalanx are removed. The attachment of the short flexor tendon is preserved. Joint motion will increase and joint function is preserved. INDICATIONS: Hallux rigidus stage 2-3 according to the Vanore classification. Salvage procedure for failed arthroplasty of the first metatarsophalangeal joint. CONTRAINDICATIONS: Hallux rigidus stage 4 according to the Vanore classification. Severe elevatus position of first ray. SURGICAL TECHNIQUE: Surgery with tourniquet is preferred. Dorsomedial skin incision. Longitudinal incision of the capsule. Removal of medial and lateral osteophytes both from the metatarsal and the proximal phalanx. Release of the sesamoids. Removal of the dorsal osteophytes from the metatarsal head and the proximal phalanx in a 45 degrees angle. Intraoperative dorsiflexion should be at least 75 degrees. Hourglass tightening of the capsule at joint level. POSTOPERATIVE MANAGEMENT: Active and passive exercises immediately after surgery. Full weight bearing is allowed. Tape in forefoot pronation if marked relieving posture. RESULTS: Follow-up study of 162 patients. Follow-up period at least 2 years (24 38 months). Increase in dorsiflexion by 27 degrees. 80% of the patients temporarily showed pain at the sesamoids. Twelve patients with progression of the osteoarthritis underwent implant arthroplasty of the first metatarsophalangeal joint (n = 10) or a fusion of the joint (n = 2) as salvage surgery. Fusion or implant arthroplasty of the first metatarsophalangeal joint could be avoided primarily. In comparison to the relevant literature the results are superior to a cheilectomy. PMID- 19137397 TI - [Treatment strategies for chronic glenoid defects following anterior and posterior shoulder dislocation]. AB - OBJECTIVE: Restoration of a stable, pain-free and functional shoulder in chronic glenoid defects following anterior or posterior shoulder dislocations. INDICATIONS: Anterior glenoid defect: all recurrent or persistent shoulder instabilities in association with chronic glenoid lesions. Posterior glenoid defect: all recurrent or persistent postreposition shoulder instabilities with chronic osseous glenoid defects. CONTRAINDICATIONS: Brachial plexus injury. Poor glenoid bone stock. SURGICAL TECHNIQUE: Anterior glenoid defect: exposition of the glenoid through a deltopectoral approach. Glenoid reconstruction by autologous iliac crest graft or coracoid transfer, in cases with progressive joint destruction in combination with shoulder arthroplasty. Posterior glenoid defect: exposition of the glenoid through a modified Brodsky approach from posterolateral. Exposure of the posterior capsule between infraspinatus and teres minor muscles, medial capsulotomy, glenoid reconstruction with auto- or allograft, normally by screw osteosynthesis. Treatment of the often accompanying anterior humeral head defect (reverse Hill-Sachs defect) by transposition of the lesser tubercle (modified from McLaughlin), defect coverage by auto- or allograft, or hemiarthroplasty through an anterior approach. In cases of persisting instability reconstruction of the glenoid defect with autologous graft and, if necessary, by shoulder arthroplasty. POSTOPERATIVE MANAGEMENT: To preserve reconstructed anatomy, a Gilchrist sling is required in anterior reconstructions for 4-6 weeks. For postoperative treatment of posterior defects a thorax abduction splint is recommended for 6 weeks. Active-assisted reduced range of motion exercise is provided under physiotherapeutic guidance according to the individual pathology. RESULTS: Clinical results following open surgery of chronic glenoid lesions in shoulder instability differ from the treatment results in acute fractures because of the often accompanying large rotator cuff tears, bad bone quality and frequently large defect size. In the hands of experienced shoulder surgeons, however, favorable results can be achieved with modern implants, leading to decisive improvement in patients' quality of life. PMID- 19137398 TI - [Percutaneous fusion technique on the thoracolumbar spine with the Expedium LIS]. AB - OBJECTIVE: Fusion can be done from the thoracic spine up to the sacrum. A cannulated Expedium screw as well as the rod can be placed percutaneously. This minimally invasive approach creates only a minor muscular trauma. INDICATIONS: Osteochondrosis of the lumbar and thoracic spine. Spondylolisthesis grade I-III according to Meyerding. Instability after nucleotomy. Type A and B fractures according to Magerl. Spondylodiscitis. Flexible scoliosis of the lumbar and thoracic spine. CONTRAINDICATIONS: Osteoporosis (t score < -1.5). Spondylolisthesis grade IV according to Meyerding. SURGICAL TECHNIQUE: The patient should be positioned prone, lying flat on the table. Use of two C-arms, one in anteroposterior and the other in lateral view. Marking of the entry point of the pedicle on the skin. Short skin incision (15 mm). Insertion of the Jamshidi needle into the pedicle. A guide wire should be inserted. Removal of the needle and start with the dilatation system, first the small one and then the big one. Preparation of the pedicle with a tap and insertion of the pedicle screw with the open and closed extension sleeves. Measurement of the rod length and insertion of the rod. Fixation of the rod with screw and removal of the extension sleeves. POSTOPERATIVE MANAGEMENT: The patient should be mobilized on the same day or the 1st day after surgery with or without a corset. RESULTS: From 2005 to 2006, 72 patients (45 female, 27 male) with a mean age of 44 years (35-73 years) were treated using this technique. Follow-up examinations were performed 3, 6, 12, and 24 months after surgery. The mean follow-up was 14 months (3-24 months). Fusion rate was evaluated by computed tomography scan or X-ray. Implant-related complications were not observed. PMID- 19137399 TI - [Use of vacuum-assisted closure therapy for the conditioning of soft-tissue defects]. AB - THE PROBLEM: Acute or chronic wounds requiring temporary wound coverage and closure. Soft-tissue defects unsuitable for any type of wound closure. Soft tissue defects associated with infection. THE SOLUTION: Temporary or definite wound closure according to a general concept for treatment of acute or chronic wounds. Support of wound healing through enhanced cell proliferation and formation of granulation tissue. Improvement of local blood circulation and eradication of infection. SURGICAL TECHNIQUE: Surgical debridement. Temporary wound closure with vacuum-assisted closure therapy. Second look with change of vacuum-assisted closure systems, irrigation and debridement procedures. Definitive wound closure through - vacuum-aided wound reduction and secondary suture or - mesh grafting or - debridement and flap coverage. RESULTS: Embedded in the general concept of treatment algorithm for acute or chronic wounds, vacuum assisted wound closure is part of the therapy. Vacuum therapy facilitates direct wound closure or improves wound conditioning to assist further surgical procedures. PMID- 19137400 TI - The mini-incision mid-vastus approach for total knee arthroplasty. AB - OBJECTIVE: Rapid functional recovery and improved range of motion after total knee arthroplasty (TKA) without compromising implant position. INDICATIONS: Osteoarthritis of the knee requiring TKA. CONTRAINDICATIONS: Preoperative flexion < 80 degrees. Flexion contracture > 20 degrees. Body mass index > 40 kg/m(2). Fixed valgus deformity > 15 degrees. Relative: previous open surgery on the knee; systematic steroids (skin fragility); tall muscular males. SURGICAL TECHNIQUE: Straight skin incision over the medial third of the patella from 2 cm proximal to the patella, and then to the level of the tibial tuberosity. Extension of capsular incision 2 cm into the vastus medialis muscle from a point 2 cm proximal to the patella. Deep incision around the medial border of the patella and distally to the level of the tibial tuberosity. The patella is displaced laterally but is not everted in flexion. Knee flexion and extension as necessary to move the soft-tissue surgical window for proximal or distal exposure. Hyperflexion of the knee only for insertion of the tibial component. POSTOPERATIVE MANAGEMENT: Knee flexion exercises extension/flexion 0-0-70 degrees using continuous passive motion the day after surgery. Weight bearing to tolerance allowed at 1st day after surgery (walker, two crutches). Thrombosis prophylaxis. RESULTS: 69 patients with 74 TKAs done through the mini-incision mid vastus approach were available with a minimum 1-year follow-up. A control group was evaluated retrospectively including 52 patients with 57 total knee replacements in which a standard medial parapatellar arthrotomy with patella eversion was used. At all clinical evaluations flexion and the ability to climb stairs were significantly superior in the mid-vastus group indicating a faster recovery and return to functional activities. There were no complications and the radiographic evaluation found no implant or limb malalignment, or signs of early loosening. PMID- 19137401 TI - Development and testing of an optimized method for DNA-based identification of jaguar (Panthera onca) and puma (Puma concolor) faecal samples for use in ecological and genetic studies. AB - The elusive nature and endangered status of most carnivore species imply that efficient approaches for their non-invasive sampling are required to allow for genetic and ecological studies. Faecal samples are a major potential source of information, and reliable approaches are needed to foster their application in this field, particularly in areas where few studies have been conducted. A major obstacle to the reliable use of faecal samples is their uncertain species-level identification in the field, an issue that can be addressed with DNA-based assays. In this study we describe a sequence-based approach that efficiently distinguishes jaguar versus puma scats, and that presents several desirable properties: (1) considerably high amplification and sequencing rates; (2) multiple diagnostic sites reliably differentiating the two focal species; (3) high information content that allows for future application in other carnivores; (4) no evidence of amplification of prey DNA; and (5) no evidence of amplification of a nuclear mitochondrial DNA insertion known to occur in the jaguar. We demonstrate the reliability and usefulness of this approach by evaluating 55 field-collected samples from four locations in the highly fragmented Atlantic Forest biome of Brazil and Argentina, and document the presence of one or both of these endangered felids in each of these areas. PMID- 19137403 TI - Biodegradation of acetonitrile by adapted biofilm in a membrane-aerated biofilm reactor. AB - A membrane-aerated biofilm reactor (MABR) was developed to degrade acetonitrile (ACN) in aqueous solutions. The reactor was seeded with an adapted activated sludge consortium as the inoculum and operated under step increases in ACN loading rate through increasing ACN concentrations in the influent. Initially, the MABR started at a moderate selection pressure, with a hydraulic retention time of 16 h, a recirculation rate of 8 cm/s and a starting ACN concentration of 250 mg/l to boost the growth of the biofilm mass on the membrane and to avoid its loss by hydraulic washout. The step increase in the influent ACN concentration was implemented once ACN concentration in the effluent showed almost complete removal in each stage. The specific ACN degradation rate achieved the highest at the loading rate of 101.1 mg ACN/g-VSS h (VSS, volatile suspended solids) and then declined with the further increases in the influent ACN concentration, attributed to the substrate inhibition effect. The adapted membrane-aerated biofilm was capable of completely removing ACN at the removal capacity of up to 21.1 g ACN/m(2) day, and generated negligible amount of suspended sludge in the effluent. Batch incubation experiments also demonstrated that the ACN-degrading biofilm can degrade other organonitriles, such as acrylonitrile and benzonitrile as well. Denaturing gradient gel electrophoresis studies showed that the ACN degrading biofilms contained a stable microbial population with a low diversity of sequence of community 16S rRNA gene fragments. Specific oxygen utilization rates were found to increase with the increases in the biofilm thickness, suggesting that the biofilm formation process can enhance the metabolic degradation efficiency towards ACN in the MABR. The study contributes to a better understanding in microbial adaptation in a MABR for biodegradation of ACN. It also highlights the potential benefits in using MABRs for biodegradation of organonitrile contaminants in industrial wastewater. PMID- 19137402 TI - Arsenic contamination in groundwater in the Southeast Asia region. AB - The adverse impact of groundwater contaminated with arsenic (As) on humans has been reported worldwide, particularly in Asian countries. In this study, we present an overview of the As crisis in the Southeast Asian region where groundwater is contaminated with naturally occurring As and where contamination has become more widespread in recent years. In this region more than 100 million people are estimated to be at risk from groundwater As contamination, and some 700,000 people are known so far to have been affected by As-related diseases. Despite investments exceeding many millions of dollars, there are still substantial knowledge gaps about the prevalence and impact of As, notably in its epidemiology, temporal variations, social factors, patient identification, treatment, etc. Arsenic-affected people in the affected regions also face serious social problems. Of major concern is the fact that many researchers from different countries have been conducting research in SE Asia region but with a lack of coordination, thus duplicating their work. There is an urgent need to coordinate these various studies to ensure better delivery of research outcomes. Further research is needed to improve field testing and monitoring of drinking water sources, and to develop new treatments for chronic As toxicity and new sources of safe drinking water. PMID- 19137404 TI - Biological sulphate reduction using food industry wastes as carbon sources. AB - Biological treatment with dissimilatory sulphate-reducing bacteria has been considered the most promising alternative for decontamination of sulphate rich effluents. These wastewaters are usually deficient in electron donors and require their external addition to achieve complete sulphate reduction. The aim of the present study was to investigate the possibility of using food industry wastes (a waste from the wine industry and cheese whey) as carbon sources for dissimilatory sulphate-reducing bacteria. The results show that these wastes can be efficiently used by these bacteria provided that calcite tailing is present as a neutralizing and buffer material. A 95 and 50 % sulphate reduction was achieved within 20 days of experiment by a consortium of dissimilatory sulphate-reducing bacteria grown on media containing waste from the wine industry or cheese whey respectively. Identification of the dissimilatory sulphate-reducing bacteria community using the dsr gene revealed the presence of the species Desulfovibrio fructosovorans, Desulfovibrio aminophilus and Desulfovibrio desulfuricans. The findings of the present study emphasise the potential of using wastes from the wine industry as carbon source for dissimilatory sulphate-reducing bacteria, combined with calcite tailing, in the development of cost effective and environmentally friendly bioremediation processes. PMID- 19137405 TI - The nuclear genome of the phytoseiid Metaseiulus occidentalis (Acari: Phytoseiidae) is among the smallest known in arthropods. AB - The genome size of the phytoseiid Metaseiulus (=Typhlodromus or Galendromus) occidentalis (Nesbitt) needs to be estimated before the whole nuclear genome can be sequenced. Two different procedures were used to estimate the genome size of M. occidentalis; (1) flow cytometry (Marescalchi et al. in Genome 33:789-793, 1990) and (2) quantitative real-time PCR (qRT-PCR) (Wilhelm et al. in Nucleic Acids Res 31:e56, 2003). Fluorescence intensity of propidium iodide-stained nuclei of M. occidentalis was measured by flow cytometry using females, males, and eggs. Only the eggs yielded peaks, which ranged in size from 35 to 160 Mb, with a tall peak of 140 Mb in 1-day-old eggs and 65 Mb in 2-day-old eggs, respectively. However, the peaks are broad and do not provide an accurate estimate. The qRT-PCR procedure required single-copy nuclear gene sequences from this phytoseiid. This was accomplished by designing degenerate primers, amplifying the Actin and EF1alpha sequences from M. occidentalis, and then designing M. occidentalis-specific primers that amplified a unique sequence. The standard qRT-PCR protocol was inefficient and amplification failed frequently, so we developed a high-fidelity qRT-PCR protocol, which utilizes a mix of two DNA polymerases (Taq and a proof-reading Tgo or ACCUZYME) to consistently amplify sequences. This allowed us to estimate the nuclear genome size of M. occidentalis as 88-90 +/- 5 Mb. When compared to other arthropod genomes, this appears to be very small. PMID- 19137407 TI - Molecular Diversity. Editorial. PMID- 19137406 TI - Heavy metal inputs evolution to an urban hypertrophic coastal lagoon, Rodrigo De Freitas Lagoon, Rio De Janeiro, Brazil. AB - This work discusses the temporal variation of metal concentrations in a hypertrophic coastal lagoon located in the metropolitan area of Rio de Janeiro (Brazil). The lagoon watershed includes one of the mostly densely urbanized areas of the city but without industrial activities. Six sediment cores were collected in the lagoon between May and July 2003 and analyzed for the concentration of metals (Fe, Al, Mn, Zn, Pb, Cu, Cr, and Ni). Typical sedimentation rate was calculated as being 0.75 cm year( - 1) and was uniform for at least the past 70 years. Therefore, the alterations in the dynamics of the lagoon caused by changes in its watershed were clearly indicated in sediment cores. The construction of an artificial canal to the sea and the increasing urbanization and soil use changes were the major factors affecting metal accumulation in the lagoon sediments. Metals typical of anthropogenic urban sources (Pb, Zn, and Cu) showed increasing loads following urbanization. PMID- 19137408 TI - Monthly changes in body condition scores and internal parasite prevalence in Nguni, Bonsmara and Angus steers raised on sweetveld. AB - The objective of the study was to determine monthly changes in body condition scores, body weights and on the prevalence of internal parasites in Nguni, Bonsmara and Angus steers raised on sweetveld. Body condition scores (BCS), body weights (BW), packed cell volume (PCV) and monthly faecal egg counts (FEC) were determined in 25 Nguni, 15 Bonsmara and 15 Angus steers. Nguni and Bonsmara steers maintained their body condition throughout the study, while the Angus lost condition. The Nguni had the highest PCV levels. The PCV levels tended to decline during the rainy season for all the breeds. The FEC were generally low. Fasciola spp. and strongyle eggs were found in 16.3 and 54.6% of the collected faecal samples, respectively. Of the three breeds, Nguni steers had the lowest parasite infestation levels, with the Bonsmara being more susceptible than the other two breeds. Generally, the egg counts observed throughout the study were low. PMID- 19137409 TI - A comparison of sodium bicarbonate infusion versus normal saline infusion and its combination with oral acetazolamide for prevention of contrast-induced nephropathy: a randomized, double-blind trial. AB - BACKGROUND: Contrast-induced nephropathy (CIN) is commonly encountered. Because the therapy of choice for prevention of CIN is controversial, in this study we compared the preventive efficacy of bicarbonate (Bi) infusion in dextrose water versus normal saline (NLS) infusion alone or in combination with oral acetazolamide (AZ). METHODS: In a double-blind and randomized clinical trial, all patients undergoing coronary angiography or percutaneous coronary intervention received NLS (NLS group), its combination with AZ (AZ group) or infusion of Bi (Bi group) before the procedures. RIFLE (risk of renal failure, injury to the kidney, failure of kidney function, loss of kidney function, and end-stage renal disease) criteria were used to define CIN-associated acute kidney injury (AKI). RESULTS: The risk of AKI in CIN was significantly lower in the Bi and AZ groups than in the NLS group (P 0.05). CONCLUSIONS: It seems that both Bi and AZ reduce the risk of CIN-related AKI, and close monitoring of serum potassium is needed during bicarbonate infusion. PMID- 19137410 TI - Performance of the Cockcroft-Gault and MDRD equations in adult Nigerians with chronic kidney disease. AB - BACKGROUND: Estimation of the glomerular filtration rate (GFR) is required in the assessment of patients with chronic kidney disease (CKD) in order to provide information regarding the functional status of the kidneys. Current guidelines advocate the use of prediction equations, such as the Cockcroft-Gault (CG) formula and the Modification of Diet in Renal Disease (MDRD) study-derived equations, over clearance of endogenous creatinine (Ccr) in achieving this aim. We were interested in knowing the accuracy of these equations in predicting the GFR in adult Nigerians with CKD. METHODS: We conducted a review of records of patients who were evaluated for CKD at the Nephrology Clinic of the Jos University Teaching Hospital between 2001 and 2003. We compared the CG and MDRD equations against the Ccr in predicting the GFR in 130 patients (88 males and 42 females) with CKD. RESULTS: The means +/- standard deviation (SD) for the measured and predicted GFR by the CG and MDRD equations were similar (17.6 +/- 25.8 ml/min, 19.9 +/- 24.0 ml/min and 21.5 +/- 28.2 ml/min, respectively; analysis of variance [ANOVA], F = 0.68, P = 0.5). The mean difference between CG and Ccr was -2.2 +/- 14.8 ml/min, with discordance at Ccr values >25 ml/min. The mean difference between MDRD and Ccr was -3.9 +/- 18.1 ml/min, with discordance at Ccr values >40 ml/min. CONCLUSION: The CG and MDRD equations provide reliable alternatives to measured Ccr in the estimation of the GFR in Nigerian patients with CKD. PMID- 19137411 TI - Towards efficient hydrogen production: the impact of antenna size and external factors on electron transport dynamics in Synechocystis PCC 6803. AB - Three Synechocystis PCC 6803 strains with different levels of phycobilisome antenna-deficiency have been investigated for their impact on photosynthetic electron transport and response to environmental factors (i.e. light-quality, quantity and composition of growth media). Oxygen yield and P(700) reduction kinetic measurements showed enhanced linear electron transport rates-especially under photoautotrophic conditions-with impaired antenna-size, starting from wild type (WT) (full antenna) over DeltaapcE- (phycobilisomes functionally dissociated) and Olive (lacking phycocyanin) up to the PAL mutant (lacking the whole phycobilisome). In contrast to mixotrophic conditions (up to 80% contribution), cyclic electron transport plays only a minor role (below 10%) under photoautotrophic conditions for all the strains, while linear electron transport increased up to 5.5-fold from WT to PAL mutant. The minor contribution of the cyclic electron transport was proportionally increased with the linear one in the DeltaapcE and Olive mutant, but was not altered in the PAL mutant, indicating that upregulation of the linear route does not have to be correlated with downregulation of the cyclic electron transport. Antenna-deficiency involves higher linear electron transport rates by tuning the PS2/PS1 ratio from 1:5 in WT up to 1:1 in the PAL mutant. While state transitions were observed only in the WT and Olive mutant, a further ~30% increase in the PS2/PS1 ratio was achieved in all the strains by long-term adaptation to far red light (720 nm). These results are discussed in the context of using these cells for future H(2) production in direct combination with the photosynthetic electron transport and suggest both Olive and PAL as potential candidates for future manipulations toward this goal. In conclusion, the highest rates can be expected if mutants deficient in phycobilisome antennas are grown under photoautotrophic conditions in combination with uncoupling of electron transport and an illumination which excites preferably PS1. PMID- 19137412 TI - Salinity affects the photoacclimation of Chlamydomonas raudensis Ettl UWO241. AB - Chlamydomonas raudensis Ettl UWO241, a natural variant of C. raudensis, is deficient in state transitions. Its habitat, the deepest layer of Lake Bonney in Antarctica, features low irradiance, low temperature, and high salinity. Although psychrophily and low-light acclimation of this green alga has been described, very little information is available on the effect of salinity. Here, we demonstrate that this psychrophile is halotolerant, not halophilic, and it shows energy redistribution between photosystem I and II based on energy spillover under low-salt conditions. Furthermore, we revealed that C. raudensis exhibits higher non-photochemical quenching in comparison with the mesophile Chlamydomonas reinhardtii, when grown with low-salt, which is due to the lower proton conductivity across the thylakoid membrane. Significance of the C. raudensis UWO241 traits found in the low salinity culture are implicated with their natural habitats, including the high salinity and extremely stable light environments. PMID- 19137414 TI - International public health law: not so much WHO as why, and not enough WHO and why not? AB - To state the obvious, "health matters", but health (or its equitable enjoyment) is neither simple nor easy. Public health in particular, which encompasses a broad collection of complex and multidisciplinary activities which are critical to the wellbeing and security of individuals, populations and nations, is a difficult milieu to master effectively. In fact, despite the vital importance of public health, there is a relative dearth of ethico-legal norms tailored for, and directed at, the public health sector, particularly at the international level. This is a state of affairs which is no longer tenable in the global environment. This article argues that public health promotion is a moral duty, and that international actors are key stakeholders upon whom this duty falls. In particular, the World Health Organization bears a heavy responsibility in this regard. The article claims that better health can and must be better promoted through a more robust interpretation of the WHO's role, arguing that neither the WHO nor international law have yet played their necessary part in promoting health for all. PMID- 19137416 TI - Low concentration of GA activates a preconditioning response in HepG2 cells during oxidative stress-roles of Hsp90 and vimentin. AB - Oxidative stress can be a significant cause of cell death and apoptosis. We performed studies in HepG2 cells to explore whether prior exposure to oxidative stress ("oxidative preconditioning") and geldanamycin (GA) treatment can protect the cell from damage caused by subsequent oxidative insults. The cells were treated with 10 nM GA for 24 h before oxidative stress. Oxidative preconditioning was achieved by 2 h exposures to H(2)O(2) (50 microM) separated by a 10-h recovery period in normal culture medium. Oxidative stress was induced by exposure to 500 microM H(2)O(2) for 24 h. The effects of GA and oxidative preconditioning were investigated on the formation of Hsp90, vimentin, insoluble vimentin aggregates, and cleavage of vimentin in a cell culture model of oxidative stress. GA treatment leads to enhanced expression of Hsp90 and vimentin and to inhibition of vimentin protein aggregation. Similar results were obtained by oxidative preconditioning. It is confirmed that low concentrations of GA protected HepG2 cells from subsequent oxidative stress by increasing the levels of Hsp90 and by alleviating the extent of cell apoptosis induced by oxidative stress, which is similar to oxidative preconditioning. However, in contrast to preconditioning, GA treatment obviously changed binding activity of Hsp90 to vimentin cleavages. All the above indicated that low concentrations of GA treatment triggered cell protection from oxidative stress. Both the level of Hsp90 and its ability to bind with vimentin were changed by low concentrations of GA and might contribute to oxidative stress protection. PMID- 19137419 TI - Changing household composition and food security among the elderly caretakers in rural western Kenya. AB - A descriptive cross-sectional study including quantitative and qualitative methods was carried out in rural western Kenya to examine household composition and food security among the elderly caretakers. The findings indicate that the deteriorating economic conditions and HIV/AIDS pandemic have led to the increase in the number of children in need of care and the unprecedented involvement of the elderly in caretaking responsibilities. The presence of children in households of the elderly have changed household composition and introduced a different social context in which intergenerational relationships between grandparents and grandchildren have been redefined. Contrary to the past, it is no longer a desire but a routine responsibility of the elderly to provide food for the increasing number of dependant children in their households. PMID- 19137418 TI - Distribution of anthocyanins delivered from a bioadhesive black raspberry gel following topical intraoral application in normal healthy volunteers. AB - PURPOSE: Results from our oral cavity chemoprevention trial demonstrated appreciable interpatient variations regarding chemopreventive efficacy of a freeze dried black raspberry (FBR) gel. We speculated these data reflected individual patient-related differences in absorption, target tissue uptake and local compound metabolism of key FBR compounds (anthocyanins). Accordingly, this study assessed the distribution of anthocyanins from the 10% (w/w) FBR gel in saliva, oral tissues and plasma. METHODS: Human subject participation entailed collection of: (1) saliva, tissue and plasma (5 min following gel application, keratinized tissues), (2) saliva and plasma (5 min after sublingual gel application), (3) saliva and plasma at 1, 2, and 4 h post gel application (keratinized tissues), and (4) saliva (cyanidin 3-rutinoside incubations). Levels of FBR anthocyanins in the respective samples were analyzed by LC/MS/MS. RESULTS: Our data show: significantly higher anthocyanin levels in saliva and oral tissues relative to matched plasma samples, marked donor-specific variations in anthocyanin uptake, sustainability of anthocyanins at the target site, pH affects anthocyanin penetration and intraoral anthocyanin decomposition and/or metabolism. CONCLUSIONS: No previous oral cavity chemoprevention trials evaluated compound distribution at the treatment site. Our data, which demonstrate a local delivery-derived pharmacologic advantage, provide insights which could advance oral cavity chemoprevention strategies. PMID- 19137420 TI - Analyzing multiple spike trains with nonparametric Granger causality. AB - Simultaneous recordings of spike trains from multiple single neurons are becoming commonplace. Understanding the interaction patterns among these spike trains remains a key research area. A question of interest is the evaluation of information flow between neurons through the analysis of whether one spike train exerts causal influence on another. For continuous-valued time series data, Granger causality has proven an effective method for this purpose. However, the basis for Granger causality estimation is autoregressive data modeling, which is not directly applicable to spike trains. Various filtering options distort the properties of spike trains as point processes. Here we propose a new nonparametric approach to estimate Granger causality directly from the Fourier transforms of spike train data. We validate the method on synthetic spike trains generated by model networks of neurons with known connectivity patterns and then apply it to neurons simultaneously recorded from the thalamus and the primary somatosensory cortex of a squirrel monkey undergoing tactile stimulation. PMID- 19137421 TI - Genetic characterization of an H5N1 avian influenza virus with neurovirulence in ducks. AB - In this report, an H5N1 avian influenza virus, A/duck/Hubei/hangmei01/2006, which could lead to acute disease including neurovirulence and mortality in ducks, was isolated in brains of domestic ducks in spring of 2006. Molecular characterization of the genes revealed that this virus harbored the common characteristics of a highly pathogenic avian influenza (HPAI). Phylogenetic analyses demonstrated that this virus was a member of the Fujian-like virus sublineage. All eight genes except NA and PB2 had the closest genetic relatives to the human influenza virus A/China/GD01/2006. It might indicate that the virus A/duck/Hubei/hangmei01/2006 originated from southern China, resulting from the wild bird migration or poultry transportation, and indicate that more surveillance upon evolution and transmission of influenza viruses in ducks was urgent. PMID- 19137423 TI - The association between observed parental emotion socialization and adolescent self-medication. AB - The current study examined the moderating influence of observed parental emotion socialization (PES) on self-medication in adolescents. Strengths of the study include the use of a newly developed observational coding system further extending the study of PES to adolescence, the use of an experience sampling method to assess the daily covariation between negative affect and substance use, and a focus on PES styles defined by the interaction of emotion-dismissing and emotion-coaching behaviors. Using multi-leveling modeling, we tested PES as a moderator of daily negative mood-substance use relation in a sample of 65 elevated-risk adolescents (48% male, 58% Caucasian, with a median age of 14). Results showed a three-way interaction between emotion-coaching PES, emotion dismissing PES and daily negative mood in predicting daily substance use. Results are discussed in terms of the importance of PES styles and their effects on self medication through compromised emotion regulation and interpersonal processes. PMID- 19137422 TI - Patients' reasons for electing to undergo total knee arthroplasty impact post operative pain severity and range of motion. AB - The present study examines the reasons cited by 103 patients for their electing to undergo total knee arthroplastic surgery and the relationship between these reasons and their post-operative pain and range of motion. Results suggest that individuals who describe different reasons for undergoing surgery vary in their post-operative recovery. Specifically, patients who cite pain as the reason they are undergoing surgery report greater levels of pain during the early post operative period. In contrast, patients who describe goals of regaining mobility or a specific activity as their reason for undergoing surgery achieve a greater range of motion during early post-operative physical therapy. Individuals who express avoidance goals for undergoing total knee arthroplasty report more severe post-operative pain at 1 and 3 months following surgery compared to patients who express approach goals. Interventions targeted towards patients reporting pre operative pain or avoidance goals may decrease subsequent post-operative pain and increase mobility. PMID- 19137424 TI - Hyperphosphorylation and accumulation of neurofilament proteins in transgenic mice with Alzheimer presenilin 1 mutation. AB - Neurofilaments (NFs) are hyperphosphorylated and accumulate in Alzheimer's disease (AD) brains. In this study, employing the transgenic mouse model, we explored the effect of presenilin 1 (PS-1) mutation on the phosphorylation and distribution of NFs. Western blot analysis showed that there was a significant increase in the phosphorylation of NF-H and NF-M subunits with a concomitant increase in phosphorylated c-Jun N-terminal protein kinase 1/2 (JNK1/2) mitogen activated protein kinase (MAPK) in hippocampus of PS-1 transgenic mice compared to that of wild-type littermates. Immunohistochemical analysis revealed that phosphorylated NFs accumulated throughout the hippocampal neurons of the transgenic mice. These findings suggest that PS-1 mutation may induce hyperphosphorylation and accumulation of NFs via a JNK1/2-involved mechanism. PMID- 19137425 TI - Low SAFB levels are associated with worse outcome in breast cancer patients. AB - The scaffold attachment factors SAFB1 and SAFB2 have been shown to function as estrogen receptor (ERalpha) co-repressors in breast cancer cells, and to affect many cellular processes such as stress response, RNA processing, and apoptosis. SAFB1 and SAFB2 have also been implicated in breast tumorigenesis: Their shared chromosomal locus at 19p13 is frequently lost in breast cancer, mutations have been identified, and overexpression results in growth inhibition. The purpose of this study was to determine SAFB1/SAFB2 protein expression in human breast tumors, to correlate their expression with either natural progression ("prognostic factor") or with response to Tamoxifen ("predictive factor"), and to analyze potential correlations with tumor characteristics. SAFB1/SAFB2 protein were measured by immunoblotting using a pan-SAFB antibody in tumor extracts from patients with long-term clinical follow-up (n = 289), a subset of whom had received no adjuvant systemic therapy after breast cancer surgery (n = 117) and another subset of whom were treated with adjuvant Tamoxifen (n = 172). SAFB levels were correlated with clinico-pathological variables and patient outcome. SAFB levels varied widely, with 25 tumors not expressing detectable levels of SAFB. SAFB expression was significantly correlated with ERalpha, HER-2, bcl-2 and with expression of other ERalpha coregulators such as SRC-3. There was no association between SAFB expression and disease free survival, however, low SAFB expression was significantly associated with worse overall survival in patients who did not receive adjuvant therapy. This study shows that low SAFB protein levels predict poor prognosis of breast cancer patients, suggesting critical functions of SAFB1 and SAFB2 in breast cancer cells. PMID- 19137426 TI - Patterns of recurrence of early breast cancer according to estrogen receptor status: a therapeutic target for a quarter of a century. AB - The current therapeutic strategy in breast cancer is to identify a target, such as estrogen receptor (ER) status, for tailoring treatments. We investigated the patterns of recurrence with respect to ER status for patients treated in two randomized trials with 25 years' median follow-up. In the ER-negative subpopulations most breast cancer events occurred within the first 5-7 years after randomization, while in the ER-positive subpopulations breast cancer events were spread through 10 years. In the ER-positive subpopulation, 1 year endocrine treatment alone significantly prolonged disease-free survival (DFS) with no additional benefit observed by adding 1 year of chemotherapy. In the small ER negative subpopulation chemo-endocrine therapy had a significantly better DFS than endocrine alone or no treatment. Despite small numbers of patients, "old fashioned" treatments, and competing causes of treatment failure, the value of ER status as a target for response to adjuvant treatment is evident through prolonged follow-up. PMID- 19137427 TI - Use of a new model of transgenic mice to clarify the respective functions of thyroid hormone receptors in vivo. AB - 3,5,3'-Triido-L: -thyronine (T3) exerts pleiotropic actions on development and homeostasis mostly via its nuclear receptors, TRalpha1, TRbeta1, and TRbeta2, encoded by the THRA and THRB genes. Muouse genetics data outline the contrasting functions of THRA and THRB, and suggest that these are dictated by both the respective abundance of the receptor isoforms in a given cell type and the differences in the intrinsic properties of the receptors. The diversity of consequences of either hypothyroidism or THRA/THRB mutation is astonishing, suggesting that TR controls a large number of genes and that the repertoire of target gene differs from one tissue to another. In order to distinguish between the direct and indirect actions of TH in vivo, we use the CRE/LoxP recombination system to control the expression of a mutant TRalpha1 receptor with dominant negative properties. Ubiquitous expression of this mutation in heterozygous mice recapitulates many consequences of TH deficiency, except in tissues where TRbeta is highly expressed. PMID- 19137429 TI - Perceived church-based needs and assets for HIV/AIDS prevention in an urban Nigerian community. AB - Health intervention programs have the potential to use church-based assets to address a wide range of health issues. This study examined the assets Nigerian church members believed their churches needed, including the assets they thought they had to engage in HIV/AIDS prevention activities. Eight hundred and thirty members from 83 churches completed a questionnaire designed from forum focus group data. Respondents were males and females aged 18 years old and above, with primary, secondary, university, or vocational education, and were more likely to be married than single (never married), separated, divorced, widowed or remarried. The data revealed that church members needed access to health promotion assets, including the expertise of members who are health professionals to engage in HIV/AIDS prevention activities. We recommend an intensive HIV/AIDS prevention reorientation-training workshop for church leaders to ensure a sustainable plan to address HIV/AIDS prevention needs. PMID- 19137428 TI - EUS-guided celiac plexus neurolysis for pain due to chronic pancreatitis or pancreatic cancer pain: a meta-analysis and systematic review. AB - The objective of this study was to evaluate the efficacy of EUS-guided CPN for pain relief in patients with chronic pancreatitis and pancreatic cancer. An initial search identified 1,439 reference articles, of which 130 relevant articles were selected and reviewed. Data was extracted from 8 studies (N = 283) for EUS-guided CPN for pain due to pancreatic cancer and nine studies for chronic pancreatitis (N = 376) which met the inclusion criteria. With EUS-guided CPN, the pooled proportion of patients with pancreatic cancer that showed pain relief was 80.12% (95% CI = 74.47-85.22). In patients with pain due to chronic pancreatitis, EUS-guided CPN provided pain relief in 59.45% (95% CI = 54.51-64.30). In conclusion, EUS-guided CPN offers a safe alternative technique for pain relief in patients with chronic pancreatitis or pancreatic cancer. In patients with pain due to chronic pancreatitis, better techniques or injected materials are needed to improve the response. PMID- 19137430 TI - Neuroprotective strategies targeting apoptotic and necrotic cell death for stroke. AB - It has been a major challenge to develop effective therapeutics for stroke, a leading cause of death and serious debilitation. Intensive research in the past 15 years have implicated many regulators and the related mechanisms by which neuronal cell death is regulated. It is now clear that even a brief ischemic stroke may trigger complex cellular events that lead to both apoptotic and necrotic neuronal cell death in a progressive manner. Although efforts at developing specific chemical inhibitors for validated targets have been successful for in vitro enzymatic assays, the development of some of such inhibitors into human therapy has been often hindered by their in vivo bioavailability profile. Considerations for the ability to chemically target a cellular mechanism in manner compatible with disease targets in vivo might be emphasized early in the development process by putting a priority on identifying key targets that can be effectively targeted chemically. Thorough interrogation of cellular pathways by saturation chemical genetics may provide a novel strategy to identify multiple key molecular entities that can be targeted chemically in order to select a target suitable for the treatment of intended human diseases such as stroke. PMID- 19137431 TI - Successful treatment of severe gastrointestinal bleeding after chemotherapy in acute myeloblastic leukemia with recombinant activated factor VII : report on one case and review of other uses in acute leukemias. AB - Hemorrhage is a frequent complication in patients with acute leukemias as a result of chemotherapy-induced myelosuppression. Gastrointestinal bleeding in thrombocytopenic patients carries a high mortality. Patients are generally managed with red blood cell, platelet suspensions, and fresh frozen plasma; and sometimes with pharmacologic and endoscopic interventions. If these therapeutic measures fail, patients might be treated with hemostatic drugs, one example of which is recombinant activated factor VII (rFVIIa). This drug is recommended for all kinds of bleeding in hemophiliacs with inhibitors; it is also being used for the treatment of bleeding in thrombocytopenia and platelet function disorders. We present our 44-year-old female patient who had gastrointestinal system bleeding after remission induction therapy for acute myeloid leukemia. Thrombocytopenia was refractory to apheresis platelets; and gastrointestinal bleeding could be controlled only after the administration of a single dose (35 microg/kg, total dose 2.4 mg) of rFVIIa. Our experience indicates that rFVIIa might be a novel treatment alternative in massive bleeding in leukemic patients with thrombocytopenia or platelet function disorders. PMID- 19137432 TI - Enhancement of survivin gene downregulation and cell apoptosis by a novel combination: liposome microbubbles and ultrasound exposure. AB - Ultrasound-mediated microbubble destruction (sonoporation) is an efficient and safe nonviral technique for gene delivery. In the present work, we hypothesized that short hairpin RNA (shRNA) interference therapy targeting human Survivin gene could be transfected by the novel combination of ultrasound exposure (USE) and liposome microbubbles (LM). ShRNA vectors targeting Survivin were constructed and transfected under USE and LM conditions. The optimal transfection efficiency and cell injury were compared with those of polyethylenimine (PEI)-mediated transfection in different cancer cell lines (HeLa, HepG2, Ishikawa, MCF-7, and B16-F10). The effects of gene downregulation and cell apoptosis were further investigated. The results indicated that P + USE + LM group could significantly increase the gene expression as compared with plasmid group, plasmid + USE group, plasmid + LM group (P < 0.001). The transfection efficiency of the novel combination was nearly equal to PEI-mediated transfection in some cancer cell lines while the cell viability did not decrease markedly. Reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis also confirmed that Survivin mRNA and protein expression could be knocked down significantly by shRNA transfection under USE and LM condition (P < 0.001). This is the first study to verify the role of shRNA therapy in vitro with novel combination of USE and LM. We concluded that this nonviral technique would be valuable in the gene transfection of shRNA and Survivin gene downregulation would lead to apparent cell apoptosis. PMID- 19137433 TI - Enantioseparation of nonproteinogenic amino acids. AB - The enantioseparation of structurally related N-protected beta-/gamma-amino acids, beta-/gamma-amino amides, and beta-/gamma-amino nitriles by using six different commercially available chiral stationary phases (CSPs) is reported. The synthetic key step to introduce stereochemical asymmetry into all compounds is an enzymatic kinetic resolution of the racemic nitriles to the respective amino amides and/or amino acids, depending on the class of enzyme (nitrile hydratase or nitrilase) applied. The separation efficiencies of all CSPs with regard to functional groups as well as structural variations of the amino acid derivatives depicted in Fig. 1 are discussed. PMID- 19137434 TI - Successful treatment of relapsing autoimmune pancreatitis in primary Sjogren's syndrome with rituximab: report of a case and review of the literature. AB - Autoimmune pancreatitis (AIP) is a rare disorder often associated with multiple autoimmune diseases like rheumatoid arthritis, inflammatory bowel disease and Sjogren's syndrome (SS). Although knowledge of AIP has grown over the last few years, little is certain about its cause and pathogenesis. Positive immunologic markers like antinuclear antibodies (ANA) or elevated serum levels of IgG4, systemic autoimmune disease association and positive response to oral steroid therapy strongly supports the idea of autoimmune mechanisms involved in the pathogenesis of AIP. We describe the first case reported on the literature of a patient with primary SS who developed relapsing AIP to steroids but responded successfully to Rituximab (RTX) therapy. New theories about the role of B-cells activity in SS and other autoimmune diseases has encourage the use of RTX, proving tolerance and efficacy especially in extra-glandular manifestations. PMID- 19137435 TI - The first 17 amino acids of the beet necrotic yellow vein virus RNA-5-encoded p26 protein are sufficient to activate transcription in a yeast one-hybrid system. AB - The beet necrotic yellow vein virus (BNYVV) RNA-5-encoded p26 protein is involved in the accentuation of symptoms expression of infected Chenopodium quinoa plants and is capable of transcription activation (TA) in yeast. TA was previously localized within the first 55 residues of the p26 protein. Interestingly, TA did not occur when C-terminally deleted forms of p26 were used. We used a genetic screen in the yeast one-hybrid system to select restored TA from randomly generated mutants. The TA domain was found to be located within the first 17 residues. Alanine replacement of aspartic acids 11, 16, and 17 within the full length p26 prevented TA but did not impair subcellular localization and the symptom expression. PMID- 19137436 TI - A patient with primary sexual headache associated with hypnic headaches. PMID- 19137437 TI - The financial dimension of integrated behavioral/primary care. AB - There are two reasons why mental health, now more appropriately termed behavioral healthcare, is declining: (a) a lack of understanding among psychotherapists of healthcare economics, particularly the intricacies of medical cost offset, and (b) our failure as a profession to see the importance of behavioral interventions as an integral part of the healthcare system inasmuch as the nation pays for healthcare, not psychosocial care. This paper will briefly describe the rapid changes in the economics of healthcare during the past 75 years, including the post World War II enthusiastic espousal of psychotherapy by the American public which was followed by a precipitous decline as our outcomes research in behavioral care remained ignorant of financial outcomes, leaving it to the government and managed care to arbitrarily curtail escalating mental health costs. At the present time psychology is on the cusp of becoming part of the healthcare system through integrated behavioral/primary care, renewing the primacy of financial considerations such as return on investment (ROI) and medical cost offset, as well as an urgency that we avoid the mistakes that are emerging in some flawed implementations of integrated care. PMID- 19137438 TI - Acetaldehyde-induced mitochondrial dysfunction sensitizes hepatocytes to oxidative damage. AB - Acetaldehyde (Ac), the main metabolite of ethanol oxidation, is a very reactive compound involved in alcohol-induced liver damage. In the present work, we studied the effect of Ac in mitochondria functionality. Mitochondria from Wistar rats were isolated and treated with Ac. Ac decreased respiratory control by 50% which was associated with a decrease in adenosine triphosphate content (28.5%). These results suggested that Ac could be inducing changes in cell redox status. We determined protein oxidation, superoxide dismutase (SOD) activity, and glutathione ratio, indicating that Ac induced an enhanced oxidation of proteins and a decrease in SOD activity (90%) and glutathione/oxidized GSH ratio (36%). The data suggested that Ac-induced oxidative stress mediated by mitochondria dysfunction can lead to cell sensitization and to a second oxidative challenge. We pretreated hepatocytes with Ac followed by treatment with antimycin A, and this experiment revealed a noticeable decrease in cell viability, determined by neutral red assay, in comparison with cells treated with Ac alone. Our data demonstrate that Ac impairs mitochondria functionality generating oxidative stress that sensitizes cells to a second damaging signal contributing to the development of alcoholic liver disease. PMID- 19137440 TI - Comment on Wall et al.: "Ethical aspects of urinary diversion for women with irreparable obstetric fistulas in developing countries". PMID- 19137442 TI - Contrast-enhanced magnetic resonance angiography in carotid artery disease: does automated image registration improve image quality? AB - Contrast-enhanced magnetic resonance angiography (MRA) is a noninvasive imaging alternative to digital subtraction angiography (DSA) for patients with carotid artery disease. In DSA, image quality can be improved by shifting the mask image if the patient has moved during angiography. This study investigated whether such image registration may also help to improve the image quality of carotid MRA. Data from 370 carotid MRA examinations of patients likely to have carotid artery disease were prospectively collected. The standard nonregistered MRAs were compared to automatically linear, affine and warp registered MRA by using three image quality parameters: the vessel detection probability (VDP) in maximum intensity projection (MIP) images, contrast-to-noise ratio (CNR) in MIP images, and contrast-to-noise ratio in three-dimensional image volumes. A body shift of less than 1 mm occurred in 96.2% of cases. Analysis of variance revealed no significant influence of image registration and body shift on image quality (p > 0.05). In conclusion, standard contrast-enhanced carotid MRA usually requires no image registration to improve image quality and is generally robust against any naturally occurring body shift. PMID- 19137443 TI - Prolonged apoptosis in mitochondria-rich cells of tilapia (Oreochromis mossambicus) exposed to elevated salinity. AB - The time-course of programmed cell death (apoptosis) during reorganization of gill epithelium in salinity-stressed tilapia was analyzed using a recently developed method based on laser scanning cytometry (LSC) of dissociated gill cells. Apoptosis in mitochondria-rich cells (MRC) was distinguished from that in other cell types using Na(+)/K(+) ATPase (NKA) as a cell-specific marker. Caspase 3/7 activity in MRC, assessed using LSC and microplate assays, increased significantly starting at 6 h of salinity stress and remained elevated for at least 5 days. This time-course of apoptosis in MRC during acute salinity stress was reflected in elevated apoptotic DNA fragmentation. In parallel to induction of apoptosis, MRC showed a pronounced shift to G2 phase of the cell cycle, which is indicative of G2/M cell cycle arrest, and an increase in NKA abundance per MRC. Unlike in MRC, apoptosis was not significantly increased in other gill cell types, although there was a small transient increase in DNA fragmentation at 6 h. G2 arrest was also observed. Overall, we interpret our data as evidence for a significant role of apoptosis in the extensive reorganization of MRC populations that takes place during salinity acclimation, perhaps similar to its well established role during organismal development. PMID- 19137444 TI - Male infertility associated with adult dominant polycystic kidney disease: a case series. AB - INTRODUCTION: Although described earlier, the association of male infertility with adult dominant polycystic kidney disease (ADPKD) is quite rare and unfamiliar to some of the multidisciplinary team members caring for affected men. MATERIALS AND METHODS: Infertile men diagnosed to have ADPKD were evaluated by clinical characteristics including testis volume, as well as serum hormone levels, semen analysis, and transrectal ultrasonography (TRUS) because of low volume ejaculate. RESULTS: Semen analysis revealed low-normal volume, normal pH, and azoospermia/virtual azoospermia. Serum hormones were within the normal range. Transrectal ultrasonography demonstrated cystic dilatation of the seminal vesicles in all three men. CONCLUSION: Patients should be referred for andrological evaluation of a presentation similar to obstructive azoospermia. Their potential to achieve paternity by surgical sperm retrieval combined with assisted reproductive technology is another example of cooperation between andrologists and gynecologists. PMID- 19137446 TI - Atypical antipsychotic use in pregnancy: a retrospective review. AB - To determine the prevalence of use of atypical antipsychotics in pregnancy and describe infant outcomes associated with these exposures. A retrospective chart review of all pregnant women presenting at our medical center, from the years 1993 to 2007, was conducted. 30,092 total deliveries during that time period were identified. 16 mothers were treated with atypical antipsychotics during their pregnancy. The medical charts of the infants exposed to atypical antipsychotics during pregnancy were reviewed to determine their outcomes, including gestational age, birth weight, and malformations. Only 16 (0.053%) of the women took an atypical antipsychotic at some point during the antenatal period. Average gestational age was 37 5/7 weeks with a standard deviation of 2.54 days. Average birth weight was 3,188 g with a standard deviation of 615 g, which is appropriate for early gestational age. One (6.25%) major malformation was noted. Additional studies need to be completed regarding the effect of atypical antipsychotic use in pregnancy and outcomes related to gestational age, birth weight and malformations. PMID- 19137447 TI - Long-term psychosocial sequelae of stillbirth: phase II of a nested case-control cohort study. AB - Stillbirth is associated with increased psychological morbidity in the subsequent pregnancy and puerperium. This study aimed to assess longer-term psychological and social outcomes of stillbirth and to identify factors associated with adverse outcome. We conducted seven-year follow-up of a cohort of women who were initially assessed during and after a pregnancy subsequent to stillbirth, together with pair-matched controls. All women were living with a partner at baseline and none had live children. Measured outcomes at follow-up included depression, posttraumatic stress disorder (PTSD) and partnership breakdown. Comparison variables included social and psychological factors and, for the stillbirth group, factors relating to the lost pregnancy. There were no differences between groups in case level psychological morbidity, but significantly higher levels of PTSD symptoms persisted in stillbirth group mothers who had case level PTSD 7 years earlier. Stillbirth group mothers were more likely to have experienced subsequent partnership breakdown. In the stillbirth group such breakdown was associated with having held the stillborn infant and having had case-level PTSD. Interpretations and clinical implications of these findings are discussed. PMID- 19137448 TI - A systematic review of home-based interventions to prevent and treat postpartum depression. AB - This systematic review evaluated randomized controlled trials assessing home based psychological interventions to prevent and treat postpartum depression (PPD). Six studies meeting inclusion criteria were identified, five assessing treatments for PPD and one assessing a preventive intervention for PPD. Interventions used cognitive behavioral, psychodynamic, and non-directive counseling approaches. Of the six studies reviewed, four reported statistically significant treatment effects on PPD following the intervention. Each of the three types of psychological interventions were shown to reduce levels of maternal depression. Recommendations for future development and evaluation of home-based psychological interventions for PPD are discussed. These include identification of mediators and moderators of intervention efficacy, exploring strategies for prevention of PPD, and targeting high-risk groups. PMID- 19137449 TI - Peptide immobilized on gold particles enhances cell growth. AB - A multivalent ligand of thrombopoietin (TPO) was prepared by immobilization of mimetic peptides on gold particles. An effective peptide ligand containing cysteine was designed to enhance the growth of TPO-sensitive cells. The peptide was then immobilized on gold particles by self assembly. The multivalent ligand enhanced the growth of TPO-dependent cells and its activity was more than that of the monovalent ligand. PMID- 19137450 TI - The effects of anthelmintic treatments against gastrointestinal nematodes on the performance of breeding ewes and lambs on pasture in semi-arid Kenya. AB - The effects of anthelmintics treatments in controlling gastrointestinal nematodes in breeding ewes in a semi-arid area of Kenya were determined. The study carried out during two breeding seasons, between June 2000 and December 2001 where albendazole was administered to groups of ewes, 2 weeks before mating, 3 weeks to lambing and mid lactation indicated significantly lower nematode egg counts in treated than untreated groups of ewes. In the first breeding season, reduced rainfall resulted in pasture scarcity and weight loss in both groups of ewes through out the gestation period, but losses were higher for the untreated group. In the second season, both groups of ewes showed a steady increase in weight gain during the gestation period and post-partum, but weight gains were higher in the treated group. In lambs, weight gains at 6 weeks were higher for treated ewes than control groups, in both breeding seasons. The results of this trial indicate that anthelmintic treatments in breeding ewes in the study area are beneficial in reducing gastrointestinal nematode infections and improving performance of the ewes and their lambs. In addition to the treatments, breeding ewes should be given feed supplementation particularly during periods of pasture scarcity. PMID- 19137451 TI - Healthcare regulation as a tool for public accountability. AB - The increasing costs of healthcare delivery led to different political and administrative approaches trying to preserve the core values of the welfare state. This approach has well documented weaknesses namely with regard to healthcare rationing. The objective of this paper is to evaluate if independent healthcare regulation is an important tool with regard to the construction of fair processes for setting limits to healthcare. Methodologically the authors depart from Norman Daniels' and James Sabin's theory of accountability for reasonableness and try to determine if new regulatory models-namely independent agencies-perform better with regard to the public disclosure of the reasons and rationales of healthcare rationing. In publicly financed healthcare systems independent regulation is an important tool to assure fair and reasonable procedures of prioritising services. In accordance with the principle of public accountability, independent regulatory agencies are particularly well suited to assure publicity of the decision-making processes, relevance of the rationale involved and particularly mechanisms for challenge and dispute resolution regarding limit setting decisions. It follows that independent healthcare regulation could be regarded not only as an instrument for performance improvement but also as a tool of social justice. The authors conclude by stating that accountability for reasonableness should be regarded as a landmark of any healthcare reform. And therefore regulators have the social task of assuring that the rationales for limit-setting decisions are clearly accessible to the public. PMID- 19137452 TI - Content analysis of euthanasia policies of nursing homes in Flanders (Belgium). AB - OBJECTIVES: To describe the form and content of ethics policies on euthanasia in Flemish nursing homes and to determine the possible influence of religious affiliation on policy content. METHODS: Content analysis of euthanasia policy documents. RESULTS: Of the 737 nursing homes we contacted, 612 (83%) completed and returned the questionnaire. Of 92 (15%) nursing homes that reported to have a euthanasia policy, 85 (92%) provided a copy of their policy. Nursing homes applied the euthanasia law with additional palliative procedures and interdisciplinary deliberations. More Catholic nursing homes compared to non Catholic nursing homes did not permit euthanasia. Policies described several phases of the euthanasia care process as well as involvement of caregivers, patients, and relatives; ethical issues; support for caregivers; reporting; and procedures for handling advance directives. CONCLUSION: Our study revealed that euthanasia requests from patients are seriously considered in euthanasia policies of nursing homes, with great attention for palliative care and interdisciplinary cooperation. PMID- 19137453 TI - Antibodies to glycosylphosphatidyl-inositol anchored proteins (GPI-AP) in antithymocyte and antilymphocyte globulin: possible role for the expansion of GPI AP deficient cells in aplastic anemia. AB - Antithymocyte globulin (ATG) and antilymphocyte globulin (ALG) are currently used successfully for immunosuppressive treatment of aplastic anemia. In this study we have investigated whether commercial ATG/ALG preparations contain antibodies against glycosylphosphatidyl-inositol anchored proteins (GPI-AP), which could be responsible for emergence of GPI-deficient populations in aplastic anemia after ATG/ALG therapy. We analyzed four commercial ATG/ALG preparations by competitive binding assays using flow cytometry. Quantification was achieved by calculating the concentration of ATG/ALG required to give 50% inhibition of binding the specific fluorochrome-labeled monoclonal antibody (EC50). High concentrations of antibodies against the GPI-anchored protein CD52 were found in all preparations (Lymphoglobulin Genzyme, Thymoglobulin Genzyme, ATGAM. Pharmacia & Upjohn, and ATG-Fresenius S Fresenius). Antibodies against the GPI-anchored protein CD48 are present in significant concentrations except in the preparation ATGAM. CD16 antibodies were found in lower concentrations. We could not detect significant concentrations of antibodies against the GPI-anchored proteins CD157 and CD14. Campath-1H, a monoclonal antibody against the GPI-anchored protein CD52, has been used as immunosuppressive tool for T-cell depletion. CD52 antibodies in ATG/ALG preparations might contribute in the same way to the immunosuppressive effects in treatment of aplastic anemia. It is known that in a substantial proportion of patients with aplastic anemia GPI-deficient cells are present in a low level at diagnosis or emerge after immunosuppressive therapy. GPI-anchored antibodies in ATG/ALG preparations might lead to a relative advantage for pre-existing GPI deficient cells caused by an escape from the antibody-mediated attack. PMID- 19137454 TI - Immunology taught by lung dendritic cells. AB - Dendritic cells (DCs) are leukocytes specialised in the uptake, processing, and presentation of antigen and fundamental in regulating both innate and adaptive immune functions. They are mainly localised at the interface between body surfaces and the environment, continuously scrutinising incoming antigen for the potential threat it may represent to the organism. In the respiratory tract, DCs constitute a tightly enmeshed network, with the most prominent populations localised in the epithelium of the conducting airways and lung parenchyma. Their unique localisation enables them to continuously assess inhaled antigen, either inducing tolerance to inoffensive substances, or initiating immunity against a potentially harmful pathogen. This immunological homeostasis requires stringent control mechanisms to protect the vital and fragile gaseous exchange barrier from unrestrained and damaging inflammation, or an exaggerated immune response to an innocuous allergen, such as in allergic asthma. During DC activation, there is upregulation of co-stimulatory molecules and maturation markers, enabling DC to activate naive T cells. This activation is accompanied by chemokine and cytokine release that not only serves to amplify innate immune response, but also determines the type of effector T cell population generated. An increasing body of recent literature provides evidence that different DC subpopulations, such as myeloid DC (mDC) and plasmacytoid DC (pDC) in the lungs occupy a key position at the crossroads between tolerance and immunity. This review aims to provide the clinician and researcher with a summary of the latest insights into DC-mediated pulmonary immune regulation and its relevance for developing novel therapeutic strategies for various disease conditions such as infection, asthma, COPD, and fibrotic lung disease. PMID- 19137455 TI - Response of human oral mucosa and skin to histamine provocation: laser Doppler perfusion imaging discloses differences in the nociceptive nervous system. AB - OBJECTIVE: To investigate the existence of histamine-excitable nerve fibers in the oral mucosa and to compare the response to histamine provocation in healthy volunteers with that in a small group of patients with chronic oral pain. MATERIAL AND METHODS: Thirteen healthy volunteers and six patients suffering from chronic oral pain took part in the study. Blood perfusion was monitored in the hard palate, the tongue, and the skin of the cheek using laser Doppler perfusion imaging (Perimed; Sweden). Baseline scannings were performed, followed by 15 scannings after iontophoresis of histamine (1%). A free description of the sensations was then obtained from the participants after finishing the measurements. RESULTS: Compared to pre-histamine scanning, histamine application resulted in a considerable increase in blood perfusion in all regions (p<0.001) that was significantly higher in skin than in oral mucosa (p<0.001). There were no significant differences between the healthy volunteers and the patients regarding baseline blood flow, increased blood perfusion, or flare size after histamine provocation. The sensory impression was reported to be more persistent and intense in the skin than in the oral mucosa. No effect on mucosa could be detected by visual inspection. CONCLUSIONS: Intra-oral flare could be induced by activating histamine-excitable nerve fibers. Both duration and intensity of the flare were considerably less pronounced than in the control skin site. Histamine application was not clearly associated with itch. PMID- 19137457 TI - Working conditions and exposure to dust and bioaerosols in sisal processing factories in Tanzania. AB - Information was collected on working conditions and personal exposures to dust, bacteria, and fungi in sisal fiber processing factories in Tanzania to identify health hazards and possible needs for preventive measures. Walkthrough surveys were performed in the brushing and decorticating departments of six sisal factories. The number of departments showing adequate scores for ergonomics and work organization, physical working environment, and occupational health items were determined. Personal thoracic dust samples were collected during sisal processing and analyzed for concentration of dust (n = 24) and for fungi and bacteria (n = 32). In both departments, most items considered to be a prerequisite for a good working environment were either missing or inadequate. Ergonomic and physical hazards were observed. Repetitive strenuous tasks, awkward work postures, and high noise levels were found. Visible dust and inadequate ventilation were seen in the brushing departments, and wet floors were observed in the decortication departments. Personal protective equipment was hardly used. The arithmetic mean exposure of sisal processors was 1.2 mg thoracic dust/m(3), 43 x 10(6) bacteria/ m(3), and 2.35 x 10(6) fungal spores/m(3). The highest exposure levels were measured in the decortication departments when machines were cleaned of waste. Significant differences were found for mean thoracic dust exposure and bacteria counts between the brushing and decortication departments and the security guards. Within individual departments, there were no significant differences in exposures between the different work tasks. A linear mixed effect model of thoracic dust including department as fixed effect explained 65% of the between-worker variability for thoracic dust exposure. The study shows that workers in sisal processing in Tanzania are exposed to bioaerosols, and suitable control measures should be implemented. More exposure studies are needed in this type of industry. PMID- 19137458 TI - Estimation of protein intake using urinary urea nitrogen in patients with liver cirrhosis. AB - OBJECTIVE: Evaluation of protein intake of patients with liver cirrhosis (LC) would facilitate optimal nutritional support. However, this has never been done based on urinary urea nitrogen (UUN). The aim of this study was to determine the usefulness of the estimated protein intake (EPI) based on UUN in patients with LC. MATERIAL AND METHODS: Eighty-two patients with LC were enrolled in this study. The actual protein intake (API) was defined as the dietary protein intake (1.0 g/kg/day) plus supplementation of any enteral diets containing branched chain amino acids (BCAA). We calculated EPI from UUN using the formula [(UUN (g/day) + 0.031 x body-weight (kg)) x0.625]. We examined the correlation between EPI and API and the EPI/API ratio (EAR), and the correction based on the results. RESULTS: A significant positive correlation was found between API and EPI (r=0.72, p<0.001). The EAR in all patients was 0.82+/-0.13. EPI x 1.2 was the correction needed to adjust EAR to 1. The corrected EPI was correlated with API (r=0.704, p<0.001). The corrected EAR of all 82 patients was 0.99+/-0.16. CONCLUSIONS: EPI calculated from UUN is a useful tool for optimal nutritional support in LC patients, and our correction greatly improves EPI accuracy. PMID- 19137459 TI - Evaluation of health-related quality of life in patients with painful bladder syndrome/interstitial cystitis and the impact of four treatments on it. AB - OBJECTIVE: Painful bladder syndrome/interstitial cystitis (PBS/IC) is an inflammatory bladder disease of unknown origin. Symptoms of PBS/IC compromise patients' quality of life (QoL). This study evaluated a health-related quality of life (HRQoL) questionnaire in PBS/IC. MATERIAL AND METHODS: 151 patients with PBS/IC filled in the HRQoL questionnaire before and after the treatment. Of these, 87 patients participated in a 3-month randomized study testing intravesical dimethyl sulfoxide (DMSO) and bacille Calmette-Guerin (BCG) and 64 patients took part in a 6-month randomized study evaluating oral cyclosporine A (CyA) and pentosan polysulfate sodium (PPS). The changes in HRQoL questionnaire were evaluated with respect to the changes in global response assessment (GRA). RESULTS: The results of the HRQoL questionnaire reflected well the post-treatment GRA. Patients responding to their treatment had improved QoL. CyA treatment had more impact on emotional well-being, social functioning, activity limitation days, pain and physical capacity than PPS treatment (p<0.05). More patients responded to DMSO than BCG treatment according to GRA (p<0.01), but the results in HRQoL questionnaire were equal after DMSO and BCG treatments. CONCLUSIONS: The HRQoL questionnaire can be used in evaluating QoL in PBS/IC patients. Treatment of PBS/IC had obvious effects on QoL. PMID- 19137460 TI - Intramuscular olanzapine versus short-acting typical intramuscular antipsychotics: comparison of real-life effectiveness in the treatment of agitation. AB - OBJECTIVE: To compare the effectiveness of intramuscular (IM) olanzapine and typical IM antipsychotics in naturalistically treated acutely agitated patients with schizophrenia or acute mania. METHODS: During the acute phase, 2011 inpatients (including emergency settings) were assessed at 2, 24 and 72 h, and 7 days following initial injection and on oral antipsychotic transition. Mean change in agitation was assessed via Positive and Negative Symptom Scale-Excited Component (PANSS-EC) and Clinical Global Impressions-Severity (CGI-S) scores. Response (> or = 40% reduction in baseline PANSS-EC score) was analysed using logistic regression. RESULTS: Significantly greater decreases in PANSS-EC and CGI S scores were observed in patients receiving IM olanzapine (n = 1294) as their first injection compared with patients receiving other IM antipsychotics (n = 717) (P<0.05; 2 h: effect size 0.1); IM haloperidol treatment (all assessments, P<0.05); and IM zuclopenthixol treatment (2 h, P<0.001). Higher response rates were observed with IM olanzapine compared with other IM antipsychotics at 24 and 72 h, and 7 days (P<0.05). IM olanzapine was associated with fewer extrapyramidal side effects compared with other assessed IM antipsychotics. CONCLUSIONS: IM olanzapine provided somewhat more effective control of acute agitation than other assessed IM antipsychotics. PMID- 19137461 TI - Quantification of menstrual flow by weighing protective pads in women with normal, decreased or increased menstruation. AB - OBJECTIVE: To investigate the association between menstrual flow weight measured from modern sanitary pads (converting liquid to non-evaporating gel) and clinically assessed normal, increased or decreased menstrual flow. DESIGN: Objective method development study. SETTING: Outpatient clinic, University Hospital, Reykjavik. POPULATION: One hundred and thirteen volunteers included 26 normally menstruating adult women and 52 normally menstruating teenagers not using oral or intrauterine contraception, seven normally menstruating women using oral contraception, 17 women with clinically diagnosed menorrhagia, five women using oral contraception for clinical menorrhagia, and six teenage girls claiming heavy menstrual flow. METHODS: Menstruation length, menstrual flow weight and history of iron deficiency were assessed. During the menstruation following recruitment, all women collected their used protective pads in a hygienic manner and returned them to the laboratory for accurate weighing. MAIN OUTCOME MEASURES: Menstrual flow total weight measured in grams. RESULTS: Mean menstrual flow total weight in the 78 asymptomatic women was 51 g (median 44, range 5-144). The mean flow in 17 women clinically diagnosed with menorrhagia was 217 g (median 207, range 63-402) (p<0.0001 compared to healthy women). The seven healthy women using oral contraceptives discharged 13 g (13-19) (p=0.0004 compared with normals). Menstruation lasted < eight days in 77/78 healthy women and in 12 of 17 clinically diagnosed menorrhagic women. CONCLUSIONS: Measurement of menstrual flow total weight accurately reflects clinically assessed normal, increased and decreased flow. The method is an easy and accurate way of objectively estimating menstrual flow. PMID- 19137462 TI - Rare skin cancer: a population-based cancer registry descriptive study of 151 consecutive cases diagnosed between 1980 and 2004. AB - BACKGROUND: There are few epidemiological data available on rare skin cancer, including Merkel cell carcinoma, Paget's disease, adnexal carcinoma, and sarcoma. We conducted this study to investigate the epidemiology of rare skin cancer diagnosed in the department of Doubs from 1980 to 2004. METHODS: Data were collected from a population-based cancer registry from 1980 to 2004. Diagnosis was based on the 3(rd) edition of the International Classification of Diseases for Oncology. The incidence rates were standardized on world population. RESULTS: One hundred and fifty one patients were investigated (88 women and 63 men). Median age for the diagnosed disease was 63 years. The standardized incidence rate was 0.82/100 000 person-year (95% CI = 0.68-0.96) and increased from 0.25 in 1980-1984 to 1.50 in 2000-2004. Fifty nine cases (39%) were sarcomas, 35 (23%) adnexal carcinomas, 27 (18%) Merkel cell carcinoma and 27 (18%) Paget's disease. The standardized incidence rates were 0.37/100 000 (0.27-0.47) for sarcomas, 0.16 (0.10-0.22) for adnexal tumors, 0.13 (0.08-0.18) for Merkel cell carcinoma, and 0.15 (0.09-0.21) for Paget's disease. CONCLUSIONS: Our results based on a population-based cancer registry showed an increase of the standardized incidence rate for all types of rare skin tumors. These results may be useful when considering the growing interest in rare diseases in identifying risk factors and planning scientific research programmes. PMID- 19137464 TI - The doctrine of integration in psychiatry and the pre-Socratics. AB - OBJECTIVE: The notion of the integration of the mental and the physical, and of a number of other divisions, is widely promoted in psychiatry. Rene Descartes is often held responsible for divisions that appear in our profession and in our practice. The wish for integration, though, fails to recognize the way in which such divisions might be fundamental and necessary in our profession. This paper describes the endeavour in psychiatry and in Western thought more generally to cover up such divisions. It traces the drive towards integration to the origins of contemporary thought in Plato and Aristotle, in order to examine what preceded such thought. CONCLUSIONS: The pre-Socratic thinkers were able to articulate fundamental and radical divisions, an ability which has been lost since the writings of Plato and Aristotle. A reappraisal of the pre-Socratics can lead to a reconsideration of the ongoing attempt to cover over the necessary divisions which mark our clinical practice, in order to be able to value what is most fundamental in our profession. PMID- 19137465 TI - Psychiatrically impaired medical practitioners: an overview with special reference to impaired psychiatrists. AB - OBJECTIVE: This paper will define and describe impairment in medical practitioners due to mental illness, with special reference to the specialty of psychiatry, and then review the prevalence and identification of impaired practitioners. METHODS: The quantitative and qualitative evidence regarding the incidence and impact of medical practitioner impairment due to mental illness (and, to some extent, substance use), with special reference to impaired psychiatrists, is reviewed. RESULTS: Medical practitioner impairment due to mental illness has a severe impact on their lives and their families due to both the effects of the disorder and the experience of communal, professional and self stigma. CONCLUSIONS: The consequences of impairment among medical practitioners and specialist psychiatrists as well as the under-reporting of impaired practitioners is a significant problem. PMID- 19137466 TI - Psychiatrically impaired medical practitioners: better care to reduce harm and life impact, with special reference to impaired psychiatrists. AB - OBJECTIVE: The aims are to briefly review treatment outcomes for impaired practitioners, and to explore how preventive and early intervention, and the accessing of and retention within treatment systems for impaired medical practitioners, and particularly psychiatrists, could be improved to maximize the doctors' chances of full recovery and to minimize danger to self and others. METHODS: The literature on the treatment and care of medical practitioner impairment due to mental illness, and substance use, with special reference to impaired psychiatrists is briefly reviewed. The implications of deficiencies of usual clinical management of doctors impaired by mental illness and opportunities for improvement in services for them are explored, including the impact of the experience of being an impaired medical practitioner under psychiatric treatment. The roles of medical boards and advisory services are examined. RESULTS: Medical practitioner impairment due to mental illness has a severe impact on doctors' lives and the lives of their families due to both the effects of the disorder and the experience of communal, professional and self stigma and discrimination. Deficiencies in usual practice in the treatment and rehabilitation of such individuals are identified, and alternatives explored. CONCLUSIONS: Prevention, early detection, intervention, and treatment programs that are more continuous more sensitive to the needs of impaired practitioners, that are more continuous, better structured, and rehabilitation and recovery focused, may be more likely to produce a positive outcome. PMID- 19137467 TI - Suicide and Western culture. AB - OBJECTIVE: The aim of this paper is to examine the cultural roots and transmission of Western suicide and suicidal behaviour. METHOD: We explored a period of antiquity (mythical Greece-61 CE) and selected accounts of 10 prominent suicides. The precipitating circumstances were tabulated and an assessment made of the most likely attendant emotions. The same process was followed for a recent period (1994-2008), from which 10 suicides were identified. The precipitating circumstances and the attendant emotions were compared. These circumstances and emotions were then compared to statements commonly encountered in clinical practice from people demonstrating suicidal behaviour. Finally, we looked for evidence that these stories (and the response models) had entered Western culture. RESULTS: Precipitating circumstances, loss of a loved one, actual or imminent execution or imprisonment, other losses and public disgrace, and the negative emotions of shame, guilt, fear, anger, grief and sorrow were common to both historical periods. These circumstances and emotions are similar to those commonly expressed by people who have demonstrated suicidal behaviour. There was a clear record (literature, visual arts) of these stories forming part of our cultural heritage. CONCLUSION: Models of maladaptive responses to certain adverse circumstances are part of Western culture. Suicide as a response to certain circumstances and negative emotions can be traced back more than 2000 years. Cultural change will be necessary to minimize suicide. PMID- 19137469 TI - Nullandi: the first name in neurophysiology and cognitive therapy. PMID- 19137468 TI - China-Australia training on psychosocial crisis intervention: response to the earthquake disaster in Sichuan. AB - OBJECTIVE: This paper describes the AusAID-funded psychosocial training program conducted jointly by Chinese and Australian mental health facilitators in the wake of the devastating earthquake disaster in Sichuan. CONCLUSIONS: A total of 280 professional and volunteer leaders in disaster mental health response from across China took part in the national disaster mental health training program. A comprehensive, evidence-based and practical program was collaboratively developed and delivered to meet the needs of the medical professionals and volunteer mental health workers working in the field. Results of the training evaluation indicated high levels of uptake and usefulness for the participants. Further outcome assessments will be utilized at the service delivery and community levels. The training program has contributed significantly to the building of national and local capacity to deliver psychological first-aid and psychosocial response management to assist the populations who have been affected by disaster. PMID- 19137474 TI - Barton Pope Lecture. PMID- 19137475 TI - Patients with postoperative atrial fibrillation have a doubled cardiovascular mortality. AB - OBJECTIVES: To investigate the impact of postoperative AF on late mortality and cause of death in CABG patients. DESIGN: All CABG patients without preoperative AF surgically treated between January 1, 1997 and June 30, 2000 were included (N = 1419). Altogether, 419 patients (29.5%) developed postoperative AF. After a median follow-up of 8.0 years, survival data were obtained, causes of death were compared and Cox proportional hazard analysis was used to determine predictors of late mortality. RESULTS: The total mortality was 140 deaths/419 patients (33.4%) in postoperative AF patients and 191 deaths/1 000 patients (19.1%) in patients without AF. Death due to cerebral ischemia (2.6% vs. 0.5%), myocardial infarction (7.4% vs. 3.0%), sudden death (2.6% vs. 0.9%), and heart failure (6.7% vs. 2.7%) was more common among postoperative AF patients. Postoperative AF was an age independent risk indicator for late mortality with a hazard ratio (HR) of 1.56 (95% confidence interval 1.23-1.98). CONCLUSIONS: Postoperative AF is an age independent risk factor for late mortality in CABG patients, explained by an increased risk of cardiovascular death. PMID- 19137476 TI - When doctors experience their vulnerability as beneficial for the patients: a focus-group study from general practice. AB - OBJECTIVE: To describe events where doctors have experienced that their own sense of vulnerability might have been beneficial for the patient. DESIGN: Qualitative focus group study with data drawn from two group sessions. Analysis was conducted with systematic text condensation. SUBJECTS: A total of 12 GPs (five men and seven women) aged 30-68 participated. Their clinical experience ranged from one to 39 years. MAIN OUTCOME MEASURES: Analysis presented different aspects of participants' experiences of vulnerability experienced as beneficial. RESULTS: The participants generously shared stories about personal and professional vulnerability which they had perceived and sometimes disclosed to the patient. One cluster of stories dealt with situations where the doctors in some way or other had identified with the patient and his or her problem. They felt that their awareness and capacity for interpretation, creative solutions, and compassion had been enhanced through recognition. Another cluster of stories covered events where uncomfortable feelings due to uncertainty or inconsiderate behaviour sharpened the doctors' reflexivity towards their own roles in the interaction. Presenting an excuse or sharing the doubt could break the ice and make a difference. IMPLICATIONS: Vulnerability may bring strength, but must be used with caution. Our study opens towards further awareness of the vulnerability of the doctor and how it can benefit the patient in some situations. PMID- 19137478 TI - Substance Use & Misuse. Introduction. PMID- 19137477 TI - Work and sexual trajectories among African American youth. AB - The beneficial or deleterious effects of employment on youth and well-being have been highly contested. This study explores whether work influences youths' sexual risk correlates in a sample of African Americans (N = 562; 55% females; M = 14.5 years, SD = 0.6 years) followed longitudinally from adolescence to early adulthood. The study used growth curve modeling to test the association between number of hours worked and condom use, sex partners' age differences, and number of partners over time. Working a greater number of hours was associated with less condom use, with the effect varying by youths' grade point average. Working a greater number of hours was associated with older sex partners among female youth. No association was found between work and number of partners. The findings suggest that working during adolescence and early adulthood increased participants' sexual activity, thus lending some support for the work consequences perspective. The implications for future research and youth development programs are discussed. PMID- 19137479 TI - Interaction of counseling rapport and topics discussed in sessions with methadone treatment clients. AB - Therapeutic rapport between counselors and clients in drug user treatment has been shown to be an important predictor of follow-up outcomes. This naturalistic study investigated the relationship of counseling rapport to drug-related topics discussed in counseling sessions in a sample of 330 clients and nine counselors. These voluntary clients had been admitted to a private, for-profit outpatient methadone treatment in Texas between September 1995 and August 1997 and received no-fee services for a year for participation in this study. The data were gathered using forms in the TCU community treatment assessments (www.ibr.tcu.edu) that measured intake information, counseling session topics, and counselor evaluation of the client. A majority were males, Hispanic, had a pending legal status and the average age was 39. Co-occurring drug dependence for these heroin users included cocaine (38%) and alcohol (31%). The results supported the hypothesis that higher rapport would be associated with addressing clients in a more "supportive approach" that emphasized relapse prevention and strengths building while lower rapport would be associated with a punitive counseling style that stressed program rules and compliance. The influences of client background, counselor differences, and during-treatment positive urines were also examined. Although counselors differed in their general manner of dealing with clients, each also showed flexibility determined in part by client behavior (such as continued cocaine use). The findings indicate that focusing on constructive solutions is the preferred counseling approach. PMID- 19137480 TI - Retention in treatment--indicator or illusion: an essay. AB - This article poses questions about the meaning of retention in substance user treatment. Retention has become a "gold standard" for substance user-treatment programs, suggesting it has become more than an indicator of positive outcomes, but a positive treatment outcome in its own right. Retention has been studied in numerous ways and has been associated with positive treatment outcomes, but questions remain about whether it has assumed greater importance than it merits. For example, it may be an artifact of other client personality characteristics that are also associated with more positive outcomes. Conversely, it may be a function of the interaction of treatment environment and client. This article suggests a need to broaden the scope of treatment-retention studies and to compare retention rates across other medical treatment environments to see if failure to complete treatment is a function of substance user treatment or of all treatment processes as well as other types of planned change processes. PMID- 19137482 TI - Contemporary psychotherapy research: implications for substance misuse treatment and research. AB - This article provides an overview of the major findings of psychotherapy research and discusses the possible implications of these findings for substance user treatment researchers and practitioners. While the centrality of relationship for human change processes was historically understood, twentieth century research tended to see relationship and person variables as secondary to operationalized "mechanisms of action" unique to particular psychotherapies. Interestingly, recent meta-analytic investigations have uncovered the weakness of randomized controlled trials (RCT) comparison investigations that have, until recently, represented the "gold standard" for the field. There has been a resurgent interest in the "common factors" that appear to be important across many effective psychotherapies. In addition, psychiatric anthropologists have contributed important information about the problems of client noncompliance with mental health treatment that parallel quantitative investigations. Substance misuse researchers have also found that client characteristics, especially clients' readiness to engage treatment, are important to investigate. The importance of the "therapeutic alliance" and the characteristics of clients and clinicians have become central areas for study, rather than variables to be controlled or excluded. Emphasis on these factors may represent the future for research in psychotherapy and substance user treatment, especially if researchers and community practitioners can join together to overcome methodological feasibility and dissemination problems that plague effectiveness research. However, the continued attractiveness of comparative studies and treatment efficacy studies may represent longstanding epistemological assumptions and responses to economic incentives that will be difficult to challenge. PMID- 19137481 TI - Problem severity and motivation for treatment in incarcerated substance abusers. AB - Studies of community-based treatment programs for substance users document that motivation for treatment is a consistent predictor of clients remaining under treatment for a longer period of time. Recent research has replicated this in prison-based treatment programs, implying that motivation is clinically important regardless of setting. The current study examines predictors of treatment motivation using data collected from 661 male drug-involved inmates during in depth interviews that include components of the Addiction Severity Index, TCU Motivation Scale, and the Heath Services Research Instrument. Findings showed treatment motivation can be measured effectively in prison-based settings. Motivation scores were not significantly different between individuals in a prison-based treatment program and those in the general prison population. Furthermore, higher motivation for treatment scores were associated with greater levels of problem severity, suggesting that individuals with more drug-use related life problems may recognize this need and desire help for beginning long term recovery. PMID- 19137483 TI - A qualitative exploration of engagement among rural women entering substance user treatment. AB - A semistructured interview was administered to a purposive sample of 12 rural women early in substance misuser treatment in a gender specific, intensive outpatient treatment program from March 2001 through March 2003. The interview used open-ended questions about the women's state of mind before entering treatment, the experiences that helped them feel more comfortable with treatment, the experiences that caused them some discomfort in treatment, and any changes they would recommend to the program content or process in an attempt to gain understanding about factors that might facilitate a subjective comfort and engagement with the treatment experience. In approaching treatment, the women almost uniformly expressed a mixture of anxiety about the requirements of treatment and cynicism about its effectiveness. Although aware of the content and structure of the treatment program, the women focused on interpersonal experiences as the critical factors in their level of comfort once in treatment. The most frequently mentioned factors adding to comfort with the treatment experience were the welcoming, accepting, and non-judgmental attitudes of the staff; having their perceptions of their problems taken seriously by the treatment staff; being with other women who shared much of their experiences; and a respectful and generally positive attitude on the part of both the staff and the patients. The only factor that consistently caused some of the women discomfort in treatment was a difficulty in trusting that some of the other women in treatment would keep the matters discussed in group sessions confidential. The women were not completely satisfied with the program content and structure, but this seemed of very little importance to them in relation to their experience of comfort with their treatment experience. PMID- 19137484 TI - The healing of our people: substance abuse and historical trauma. AB - For the past two decades, one of the authors (Robert Morgan) has been involved in the development and implementation of culturally based outpatient, inpatient, and aftercare programs for Alaskan native and American-Indian populations in Alaska. After years of observation, it was concluded that the best efforts of our clinicians were inadequate to the task at hand, i.e., that of resolving the social and physical ills that have ravaged the Alaskan peoples since the occupation. Morgan and others sought to create a new model of diagnosis and treatment that combined the cultural strengths of the people with the technical and treatment skills of the conventional medical profession. The model was grounded in a clear understanding of the factors causing disease in the people, and that understanding came from the people themselves. Before the growth of the "healing plant" that Uncle Walter spoke of could be nurtured, it was necessary to first examine the question of cause and effect. Much of the cause is rooted in the "historical trauma" experienced by the Alaska Native People. The effects are numerous, but one of the most obvious symptoms is substance misuse. PMID- 19137485 TI - Alcohol addiction and the attachment system: an empirical study of attachment style, alexithymia, and psychiatric disorders in alcoholic inpatients. AB - This study aims at investigating alcoholic inpatients' attachment system by combining a measurement of adult attachment style (AAQ, Hazan and Shaver, 1987. Journal of Personality and Social Psychology, 52(3): 511-524) and the degree of alexithymia (BVAQ, Bermond and Vorst, 1998. Bermond-Vorst Alexithymia Questionnaire, Unpublished data). Data were collected from 101 patients (71 men, 30 women) admitted to a psychiatric hospital in Belgium for alcohol use-related problems, between September 2003 and December 2004. To investigate the research question, cluster analyses and regression analyses are performed. We found that it makes sense to distinguish three subgroups of alcoholic inpatients with different degrees of impairment of the attachment system. Our results also reveal a pattern of correspondence between the severity of psychiatric symptoms personality disorder traits (ADP-IV), anxiety (STAI), and depression (BDI-II-Nl) and the severity of the attachment system's impairment. Limitations of the study and suggestions for further research are highlighted and implications for diagnosis and treatment are discussed. PMID- 19137487 TI - Doctor's orders. 2009 legislative preview. PMID- 19137488 TI - Don't ban the ban. TMA works to preserve ban on corporate practice. PMID- 19137489 TI - The obesity threat. Fat endangers wallets, workforce. PMID- 19137490 TI - Code blues. Physicians oppose rushed ICD-10 adoption. PMID- 19137491 TI - Stemming STEMI. Physicians take on potential killer. PMID- 19137492 TI - Health information technology policy in Texas: statewide, regional, and constituency-specific initiatives. PMID- 19137493 TI - [Performance of a newly developed clip (Tumark Professional) for MRI-guided lesion localization after MRI-guided vacuum-assisted biopsy--first results]. AB - PURPOSE: Evaluation of a newly developed Clip (Tumark Professional) for MRI guided lesion localization after MRI-guided vacuum-assisted biopsy (VAB) with regard to the exactness of positioning, migration, and visibility on mammography (MG), ultrasound (US) and MR imaging (MRI). MATERIALS AND METHODS: 27 consecutive patients with 29 suspicious breast lesions detected with MRI were prospectively evaluated. The location of the lesion was determined with Tumark (Somatex, Teltow, Germany) after MRI-guided VAB. The distance between the clip and lesion was measured via MRI. The qualitative visibility of the clip was assessed by means of a 5-point scale from very good (1 point) to not visible (5 points). The analysis was performed for MG, US and MRI separately. Clip movement was measured via MG. RESULTS: 9 lesions were malignant (31%). All but one lesion (96%) were able to be localized exactly with a clip-lesion distance of < or = 10 mm. The Tumark was visible in 27 cases (93.1%) in US and in 25 cases (86.2%) in MRI. The visibility of the clip was moderate for both modalities (mean 3.2 points). Its visibility in MG was always very good (1 point). The clip position was stable at the time of short term follow-up (1 - 7 months; mean deviation 4.5 mm). CONCLUSION: Precise positioning of the Tumark Professional is usually possible. The clip is mostly visible in US. At the time of short-term follow-up, there was no relevant movement. Therefore, Tumark seems to be suitable for MRI-guided lesion localization after MRI-guided VAB of suspicious breast lesions. Further improvement of US visibility would be beneficial. PMID- 19137494 TI - [Myelolipoma versus extramedullary Hematopoiesis -- an unusual differential diagnosis in the presacral region]. PMID- 19137495 TI - A new sonographic weight formula for fetuses 80% inhibition at 100 microg/mL) with IC50 values in the range of 2-20 microg/mL. Among the isolated alkaloids, a piperidine-type alkaloid (caulophyllumine B) and three lysine-derived alkaloids (O-acetlybaptifolin, anagyrine, and lupanine) inhibited these enzymes to various extents (IC50:2.5-50 microM). N-Methylcytisine weakly inhibited CYP3A4 only (32% inhibition at 100 microM). An equimolar mixture of alkaloids exhibited a more pronounced inhibitory effect on all four enzymes as compared to the isolated alkaloids. Among the saponins, caulosides C and D inhibited CYP3A4 at the highest test concentration of 100 microM (43% and 35% inhibition, respectively). Other enzymes were not affected. This in vitro study indicates that dietary supplements containing blue cohosh may pose a risk of drug-drug interactions if taken with other drugs or herbs, metabolism of which involves CYP450 enzymes. PMID- 19137501 TI - [Tuberculosis in healthcare workers]. AB - Perception and knowledge of the TB-infection risk in healthcare workers (HCWs) changed profoundly in Germany during the past few years. Molecular epidemiological studies and a comprehensive review of the existing evidence concerning the infection risk for HCWs lead to the conclusion that TB in HCWs is often caused by infection at the workplace. In the Hamburg Fingerprint Study, 80 % of the TB cases in HCWs were caused by infections at the workplace. In a similar Dutch study 43 % of all cases were work-related. Besides of the well known risks in TB wards and laboratories, an increased risk for infection should be assumed for paramedics, in emergency rooms, for HCWs caring for the elderly or for workers with close contact to high-risk groups (homeless people, i. v. drug users, migrants from high-incidence countries). TB in a HCW working in these fields can be recognised as an occupational disease (OD) without identifying a particular source of infection. For all other HCWs, the German occupational disease law requires the identification of a source case before TB in an HCW can be accepted as an OD. Even though the proportion of work-related TB in HCWs is higher than was assumed before previously, the prevalence of latent TB infection (LTBI) is lower than expected. In an ongoing evaluation study of the interferon gamma release assay (IGRA) LTBI prevalence in HCWs is 10 %. Prevention strategies in Germany should be reconsidered in the light of these new findings. PMID- 19137502 TI - [Detection of respiratory viruses--how, when, where and why?]. AB - Respiratory viruses trigger the majority of common colds, acute respiratory illnesses in children during the cold season as well as acute exacerbations of asthma and chronic obstructive pulmonary disease. They also play a role in community acquired pneumonia. Unfortunately their detection is still difficult. The aim of this review is therefore to introduce the methods of detection and to present the current knowledge of the clinical role of respiratory viruses in different diseases. PMID- 19137503 TI - [Assessment and outcome parameters in COPD]. AB - BACKGROUND: A standard outcome parameter for pharmacological trials in COPD has not yet been defined. Therefore, it is the aim of this review to evaluate frequently used parameters for their eligibility as assessment and outcome parameters in COPD. METHODS: A review of the actual scientific literature was performed. RESULTS: It is recommended to continue to rely primarily on the FEV (1), which has been used as a primary variable in the vast majority of trials. In addition, further parameters, such as FVC and IC/TLC should be determined. If available, additional information is provided by RV/TLC, K (co), PaO (2) and PaCO (2). FEV (1) is not a surrogate parameter for dyspnoea, quality of life, and exercise tolerance, which should therefore be assessed separately. Frequency and severity of exacerbations and mortality are important outcome parameters in long term trials. Complex indices, such as the BODE index, may be superior to single variables. CONCLUSIONS: No single additional parameter has been evaluated sufficiently in order to substitute FEV (1) as the standard parameter for the assessment and outcome in COPD. PMID- 19137504 TI - [The genetics of chronic obstructive pulmonary disease]. AB - COPD is a heterogenous disease caused by the interaction of genetic susceptibility and environmental influences. The best example to support this is tobacco smoke. Although cigarette smoking is the most important aetiological factor, only up to half of chronic smokers develop significant COPD. There are three main themes within the pathogenesis of COPD: 1) imbalance between proteases and anti-proteases, 2) oxidative stress, 3) inflammation. Disparity between levels of exogeneous oxidants, e. g., tobacco smoke, and endogeneous antioxidants leads to oxidative stress which, in turn, causes an inflammatory response involving pro-inflammatory mediators. The activated inflammatory cells release further proteases and oxidants, leading to chronic inflammation and irreversible destruction of connective tissue in the lung. Individual genetic variations influence these processes in many ways. This article summarises the results of recent candidate gene studies for COPD. PMID- 19137506 TI - No effect on survival of home psychosocial intervention in a randomized study of Danish colorectal cancer patients. AB - OBJECTIVE: We investigated whether regular home visits to persons with newly diagnosed colorectal cancer influenced their overall survival and selected immune parameters. METHODS: A total of 249 Danish colorectal cancer patients undergoing abdominal surgery were randomly assigned to a control or an intervention group. The intervention group received 10 home visits from a project nurse or a medical doctor during the first 2 years after discharge. The home visits aimed at providing emotional support and information. A subgroup of 55 patients provided blood samples 3, 12 and 24 months after discharge for measurement of immune parameters. Survival was assessed 6.5-9.5 years after the first operation. RESULTS: A total of 148 patients died during follow-up. The intervention was not significantly associated with survival (p=0.68) after adjustment for Dukes' stage, radicality of the operation, age, sex, family social class and marital status. Likewise, no significant interactions were found between group and these covariates (all p > or = 0.08). In the substudy of the possible effect of the intervention on immune parameters, there were no differences between the two groups with respect to lymphocyte proliferation (all p > or = 0.078) or natural killer cell activity (all p > or = 0.33) and no consistent effect on the number of specific subsets of cells (phenotypes) during follow-up. CONCLUSION: The study failed to provide evidence that the psychosocial intervention provided as home visits significantly affected the prognosis or selected immune parameters of patients who had undergone surgery for colorectal cancer. PMID- 19137507 TI - Cancer patients' decisions about discussing Internet information with their doctors. AB - OBJECTIVE: To explore cancer patients' and caregivers' decisions regarding whether to discuss cancer-related information they found on the Internet with their doctors. METHODS: 238 participants (cancer patients and caregivers) from three online cancer communities responded to a survey about their experiences finding information on the Internet about their cancer and their reasons for discussing or not discussing that information with their doctors. The reasons were coded into mutually exclusive categories. RESULTS: Participants most frequently reported discussing information in order to be proactive in improving their health. Other reasons included appealing to the doctor as expert, wanting to become more educated, meeting a psychological or coping need, and checking up on or testing the doctor. Of the participants, caregivers were more likely than patients to cite checking up on or testing the doctor as a reason for discussing Internet information. Sixty-two percent of participants reported sometimes or never talking about Internet information that they thought was important with their doctors. Across all participants, the most frequently reported reasons for not talking about Internet information were attributions about the information and systems-related reasons. Participants who reported information attribution as a reason for not discussing it with their doctors had higher overall comfort levels with these types of discussions. CONCLUSIONS: Many factors influence cancer patients' and caregivers' decisions about discussing Internet information with their doctors. The coherence of the reasons across the communities in this study and in other studies suggests that this typology of reasons is both thorough and valid. PMID- 19137508 TI - Quality of life in the chemotherapy treatment of Spanish cancer patients: a comparison of general population norms. AB - OBJECTIVE: The SF-36 Health Survey (SF-36) is a general instrument used to assess health-related quality of life (HRQL). The purpose of this study is to describe self-reported information on health and the quality of life among cancer outpatients during chemotherapy treatment and to compare it with the baseline sample drawn from the general Spanish population. METHODS: A randomised sample of 583 cancer outpatients completed the questionnaire. Using a cross-sectional analysis of the SF-36 data, standardised scores were used to compare patients with comparative Spanish baseline data. Other health-related concerns (gender, age, site of cancer) were also analysed. RESULTS: The patients showed acceptable levels on HRQL and no significant differences were found between the oncology patients compared with the general population for any of the SF-36 domains. The older patients had scores significantly lower in physical functioning (p<0.01) and physical role (p<0.05). The results identified lower HRQL ratings among women compared with men on all the scales except general health. In relation to the location of the cancer, patients with genitourinary tumours scored statistically lower (p<0.05) on bodily pain in the SF-36 domain compared with patients with gastrointestinal tumours. CONCLUSIONS: Although more studies are needed, the SF 36 allows us to describe the HRQL of cancer outpatients during chemotherapy treatment compared with the general population. PMID- 19137509 TI - How children cope when a parent has advanced cancer. AB - OBJECTIVE: When parents are diagnosed with cancer, children experience significant distress. There is little information regarding the nature of this distress and how children cope, particularly when a parent is diagnosed with advanced cancer. This study aimed to explore how children cope, and to identify areas where there may be barriers to children accessing support to enable them to cope. METHODS: Semi-structured interviews were conducted with ill parents with advanced cancer and well parents and/or any children above the age of 7. Interviews were recorded and transcribed fully, and analysed using a constructionist grounded theory approach. RESULTS: Twenty-eight family participants were interviewed. Four major themes emerged from the data including response to diagnosis, mechanisms of coping, life changes, and positive aspects. Children described being distressed by their parents diagnosis and having concerns related to their parents and their own health. Distraction and maintaining normality were described as the dominant strategies of coping for children, and increased responsibilities and decreased social activity were considered to be the most noticeable of life changes. Parents did not recognise the impact on children to the same degree as described by children and focused on limiting the impact by maintaining normality. Positive aspects described by children and parents included strengthening of relationships and learning to value family members and the important things in life. CONCLUSIONS: Open communication within the family may lead to more effective coping and a positive experience for children whose parents have been diagnosed with advanced cancer. PMID- 19137510 TI - Carbon-based sorbents in chromatography. New achievements. AB - The newest results in the employment of carbon-based composites in various chromatographic techniques such as gas-liquid chromatography, high-performance liquid chromatography and electrically driven separation techniques for the separation, quantitative determination and identification of a wide variety of compounds in complicated matrices are compiled. The results are concisely described and critically evaluated. The future trends in the application of carbon-based compounds in the chromatographic separation methods are briefly discussed. PMID- 19137511 TI - A model for the analysis of competitive relaxation effects of manganese and iron in vivo. AB - Manganese (Mn) and iron (Fe) are both paramagnetic species that can affect magnetic resonance relaxation rates. They also share common transport systems in vivo and thus in experimental models of metal exposure their effects on relaxation rates may interact in a complex fashion. Here we present a novel model to interpret the combined effects of Mn and Fe on MRI relaxation rates. To achieve varying levels of both metals, adult rats were separated into four groups; a control group and three groups treated with weekly intravenous injections of 3 mg Mn/kg body for 14 weeks. The three treated groups were fed either a normal diet, Fe deficient or Fe enriched diet. All rats were scanned using MRI at the 14th week to measure regional water relaxation rates. Rat brains were removed at the end of the study (14th week) and dissected into regions for measurement of Mn and Fe by atomic absorption spectroscopy. For the normal diet groups, R(1) was strongly correlated with tissue Mn concentrations. However, the slopes of the linear regression fits varied significantly among different brain regions, and a simple linear model failed to explain the changes in relaxation rate when both Mn and Fe contents changed. We propose a competition model, which is based on the ability of Mn and Fe to compete in vivo for common binding sites. The combined effect of Mn and Fe on the relaxation rates is complicated and additional studies will be necessary to explain how MRI signals are affected when the levels of both metals are varied. PMID- 19137513 TI - Acquired factor VII deficiency associated with Wilms tumor. AB - We present the case of a 2-year-old female with Wilms tumor whose initial evaluation revealed a prolonged prothrombin time (PT) and normal activated partial thromboplastin time. Mixing studies demonstrated correction of the PT and the Factor VII activity was 17% in the absence of a Factor VII inhibitor. She underwent successful resection of the tumor with fresh frozen plasma support and no excessive bleeding. Post-operative testing demonstrated normal PT at 3 days and 1-month. Although acquired von Willebrand factor deficiency has a known association with Wilms tumor, paraneoplastic factor VII deficiency associated with Wilms tumor is previously unreported. PMID- 19137514 TI - Critical enhancements of MRI contrast and hyperthermic effects by dopant controlled magnetic nanoparticles. AB - Doped up: The incorporation of Zn(2+) dopants in tetrahedral sites leads to the successful magnetism tuning of spinel metal ferrite nanoparticles (see picture). (Zn(0.4)Mn(0.6))Fe(2)O(4) nanoparticles exhibit the highest magnetization value among the metal ferrite nanoparticles. Such high magnetism results in the largest MRI contrast effects (r2=860 mm(-1) s(-1)) reported to date and also huge hyperthermic effects. PMID- 19137516 TI - Dicationic sulfur analogues of N-heterocyclic silylenes and phosphenium cations. AB - DABling with sulfur: Sulfur(II) dications can be prepared using alpha-diimines to stabilize the positive charge (see scheme; DAB = diazabutadiene, Dipp = 2,6 diisopropylphenyl, OTf = CF(3)SO(3)). The bonding is best described as that of a N,N-chelated sulfur(II) dication; these species represent the first sulfur-based structural mimics of N-heterocyclic silylene compounds and phosphenium cations. PMID- 19137515 TI - Synthesis, chiral resolution and pharmacological evaluation of a 2,3 benzodiazepine-derived noncompetitive AMPA receptor antagonist. AB - The resolution of 1-(4-aminophenyl)-3,5-dihydro-3-N-ethylcarbamoyl-5-methyl-7,8 methylenedioxy-4H-2,3-benzodiazepin-4-one (R,S)-(+/-)-5 by chiral HPLC and assignment of the absolute configuration of the two enantiomers was carried out. Compound (R,S)-(+/-)-5 and its enantiomers were tested in a binding assay to evaluate their affinity for AMPA receptors. Enantiomer (S)-(-)-5 appears to be more potent than its optical antipode (R)-(+)-5. In a primary culture of rat cerebellar granule cells, which express AMPA receptors, (R,S)-(+/-)-5 and (S)-(-) 5 inhibited kainate- induced [Ca(2+)](i) increase, thus confirming the antagonism at the AMPA receptor. PMID- 19137517 TI - Crystallographic observation of dynamic gas adsorption sites and thermal expansion in a breathable fluorous metal-organic framework. AB - Playing accordion: Cooling a single crystal of a microporous fluorous metal organic framework under ambient atmosphere leads to very large breathing upon gas adsorption, during which multiple N(2) molecules are filled into channels and cages (see picture). While the framework exhibits remarkable positive thermal expansion under vacuum, a gigantic apparent negative thermal expansion takes place when the crystal is exposed to N(2) at ambient pressure. PMID- 19137518 TI - Sequential assembly of phototunable ferromagnetic ultrathin films with perpendicular magnetic anisotropy. AB - Getting organized: Assemblies of ferromagnetic FePt nanoparticles were generated with large perpendicular magnetic anisotropy by a magnetic-field-assisted layer by-layer method, and subsequently layer-by-layer films consisting of L1(0)-FePt nanoparticles and organic polymers were prepared. These films are phototunable when photochromic molecules are used as polymer layers. PMID- 19137519 TI - The synthesis and in vivo evaluation of 2',2'-difluoro KRN7000. AB - The synthesis of 2',2'-difluoro KRN7000 is described. In vivo evaluation demonstrates that this fluorinated glycolipid induces CD1d-dependent TCR activation of NKT cells, with a bias towards Th2 cytokine production. PMID- 19137520 TI - Synthesis of unprecedented scaffold diversity. AB - Building a better library: The synthesis of a library of natural-product-like small molecules with unprecedented scaffold diversity has been reported (see scheme; Ns = nosylate). The library represents a significant advancement in the capability of synthetic chemists to generate structurally diverse and complex small molecules in a rapid manner. PMID- 19137521 TI - Reactive intermediates in oxygenation reactions with mononuclear nonheme iron catalysts. AB - An advanced intermediate: A nonheme iron(IV) oxo complex [Fe(IV)(O)(bqen)(L)](n+) (bqen = N,N'-dimethyl-N,N'-bis(8-quinolyl)ethane-1,2-diamine, L = CH(3)CN or CF(3)SO(3)(-)) activates the C-H bonds of alkanes and alcohols by a hydrogen-atom abstraction mechanism. The catalytic oxidation of these species is proposed to occur through a nonheme iron(V) oxo species, with a high reactivity in oxidation reactions (see picture). PMID- 19137522 TI - Photoresponsive cyclodextrin-covered nanocontainers and their sol-gel transition induced by molecular recognition. AB - Springing the trap: Cyclodextrin-covered mesoporous silica nanoparticles with photocleavable linkers exhibit photoinduced release characteristics and a sol-gel transition that is induced by molecular recognition (see picture). Upon exposure to UV light, the guest molecules were released from the pore by removal of the CD "gatekeeper", which was linked on the surface of the silica nanoparticle through a photocleavable o-nitrobenzyl ester moiety. PMID- 19137523 TI - Silaphenylmercuric triflate catalyzed reactions: synthesis of a solid-supported mercuric salt catalyst. AB - Let it flow, let it flow: A procedure to generate the first solid-supported mercuric salt, silaphenylmercuric triflate, is described. Silaphenylmercuric triflate showed remarkable catalytic activity for an indole synthesis, furanoyne cyclization, arylyne cyclization, and tandem carbocyclizations. An efficient flow reaction system for indole synthesis and arylyne cyclization is also described (see figure). PMID- 19137524 TI - Influence of image-analysis software on quantitation of two-dimensional gel electrophoresis data. AB - Image analysis of two-dimensional gels is a crucial step in a proteomic workflow and has a direct impact on obtained qualitative and quantitative data. Since the analysis is a complex process and creates large data amounts, the use of a respective software is inevitable. There are only a few papers published addressing the issue of analysis-based variance; therefore, our aim was to highlight the discrepancy of received results when different commercially available image-tools are used for gel analysis especially in terms of comparability of the obtained outcome when the same digital image set is used. A set of six gels (three replicates per group) of real-life samples was created and examined with two different versions of PD-Quest (Bio-Rad) (version 6.1 and its update version 8.0) and with an external image-tool Delta 2D (Decodon) (version 3.6). Replicate groups were analyzed and compared with each other with regard to volume ratios of a group of significantly changed spots. The study points out significant variations among results depending on the software package used, underlining the importance of a careful investigation of post-experimental processes to receive comparable and reliable results. PMID- 19137525 TI - Identification of CpG methylation of the SNRPN gene by methylation-specific multiplex PCR. AB - In this article, we show that methylation-specific multiplex PCR (MS-multiplex PCR) is a sensitive and specific single assay for detecting CpG methylation status as well as copy number aberrations. We used MS-multiplex PCR to simultaneously amplify three sequences: the 3' ends of the SNRPN gene (for unmethylated sequences), the KRITI gene (as internal control), and the promoter of the SNRPN gene containing CpG islands (for methylated sequences) after digestion with a methylation-sensitive restriction enzyme (HhaI). We established this duplex assay for the analysis of 38 individuals with Prader-Willi syndrome, 2 individuals with Angelman syndrome, and 28 unaffected individuals. By comparing the copy number of the three regions, the methylation status and the copy number changes can be easily distinguished by MS-multiplex PCR without the need of bisulfite treatment of the DNA. The data showed that MS-multiplex PCR allows for the estimation of the methylation level by comparing the copy number aberrations of unknown samples to the standards with a known methylated status. The in-house designed MS-multiplex PCR protocol is a relatively simple, cost-effective, and highly reproducible approach as a significant strategy in clinical applications for epigenetics in a routine laboratory. PMID- 19137526 TI - The effect of aggregation on the separation performance of bacteria in capillary electrophoresis. AB - During the last 15 years, methods for the capillary electrophoretic separation of different bacteria species have been developed, which exploit their characteristic cell surface-charge to volume ratio. A special variant, the polymer-based CE of bacteria, includes a focusing step, which forces the bacteria cells to form aggregates at the beginning of the electrophoretic process, resulting in very high apparent efficiencies. Our experiments presented in this article reveal that the migration time of bacteria species in polymer-based CE increases with a growing amount of injected cells. Thus, the electrophoretic mobilities are not characteristic for the single cells of one species, but for the aggregates of the bacteria species, which are formed during the focusing process. Electrophoretic mobility (EM) data are obviously inapplicable for the identification of bacteria if the concentration of the bacteria sample solution is not constant. Fractions taken during the electrophoretic separation of different bacteria species were cultivated and tested for species purity. Interestingly, the electrophoretic bands were never pure, as all of them contained different mixtures of the injected species. We attribute this to the formation of stable mixed-species aggregates during polymer-based focusing. The mixed clusters migrate in the electric field with consistent velocity as a whole and are not separated electrophoretically. PMID- 19137528 TI - Optimization of separation and online sample concentration of N,N dimethyltryptamine and related compounds using MEKC. AB - The optimal separation conditions and online sample concentration for N,N dimethyltryptamine (DMT) and related compounds, including alpha-methyltryptamine (AMT), 5-methoxy-AMT (5-MeO-AMT), N,N-diethyltryptamine (DET), N,N dipropyltryptamine (DPT), N,N-dibutyltryptamine (DBT), N,N-diisopropyltryptamine (DiPT), 5-methoxy-DMT (5-MeO-DMT), and 5-methoxy-N,N-DiPT (5-MeO-DiPT), using micellar EKC (MEKC) with UV-absorbance detection are described. The LODs (S/N = 3) for MEKC ranged from 1.0 1.8 microg/mL. Use of online sample concentration methods, including sweeping-MEKC and cation-selective exhaustive injection-sweep MEKC (CSEI-sweep-MEKC) improved the LODs to 2.2 8.0 ng/mL and 1.3 2.7 ng/mL, respectively. In addition, the order of migration of the nine tryptamines was investigated. A urine sample, obtained by spiking urine collected from a human volunteer with DMT, was also successfully examined. PMID- 19137527 TI - Connecting humor, health, and masculinities at prostate cancer support groups. AB - OBJECTIVE: Many commentaries about men's health practices and masculinities indicate that men do not typically engage with self-health or acknowledge illness, let alone openly discuss their health concerns with other men. Prostate cancer support groups (PCSGs) appear to run contrary to such ideals, yet the factors that influence men's attendance and engagement at group meetings are poorly understood. As part of a larger PCSG study, we noticed that humor was central to many group interactions and this prompted us to examine the connections between humor, health, and masculinities. METHODS: A qualitative ethnographic design was used to direct fieldwork and conduct participant observations at the meetings of 16 PCSGs in British Columbia, Canada. Individual semi-structured interviews were completed with 54 men who attended PCSGs to better understand their perceptions about the use of humor at group meetings. RESULTS: Four themes, disarming stoicism, marking the boundaries, rekindling and reformulating men's sexuality, and when humor goes south were drawn from the analyses. Overall, humor was used to promote inclusiveness, mark the boundaries for providing and receiving mutual help, and develop masculine group norms around men's sexuality. Although there were many benefits to humor there were also some instances when well-intended banter caused discomfort for attendees. CONCLUSIONS: The importance of group leadership was central to preserving the benefits of humor, and the specificities of how humor is used at PCSGs may provide direction for clinical practice and the design of future community-based men's health promotion programs. PMID- 19137529 TI - Determination of bromate in highly saline samples using CZE with on-line transient ITP. AB - We developed CZE with direct UV detection for the determination of bromate in highly saline samples such as seawater and salts using transient ITP as an on line concentration procedure. The following optimum conditions were established: BGE, artificial seawater containing no bromide adjusted to pH 3.0; detection wavelength, 210 nm; vacuum injection period of sample, 18 s (378 nL); terminating ion solution, 600 mM sodium acetate; vacuum injection period of the terminating ion solution, 7 s (147 nL) for seawater and 12 s (252 nL) for salts; applied voltage, 7 kV with the sample inlet side as the cathode. The LOD for bromate was 30 microg/L (BrO(3) (-)-Br) with S/N of 3. The respective values of the RSD of the peak area, peak height, and migration time for bromate were 6.4, 1.5, and 0.51%. Seawater and salt samples, with bromate added, were analyzed using this method. The recovery of bromate in seawater samples was 85-105%. Linear regression equations relating area and height responses to concentration for bromate were obtained using the salt samples. PMID- 19137530 TI - Open tubular CEC in a microfluidic chip for rapid chiral recognition. AB - Chiral recognition of dansyl enantiomeric amino acids by microfluidic open tubular CEC (muOTCEC) with fluorescence detection was demonstrated. Avidin was employed as the chiral selector immobilized on the microchannel wall, which functioned as the chiral stationary phase (CSP) by physical adsorption. The condition of CSP on the glass wall was characterized using field emission SEM. Results indicated that avidin was homogenously distributed on the microchannel surface. Two parameters that played essential roles in muOTCEC for chiral recognition were investigated. Buffer pH could greatly change the amount of adsorption of avidin on the channel wall by altering the electrostatic attraction between them. Methanol, the organic additive to the running buffer, was also found significant for controlling the quality of the muOTCEC chiral separation by regulating the hydrophobic interaction between the enantiomers and the CSP. Under the optimized conditions, four dansyl racemic amino acids were then successfully separated by muOTCEC within 100 s with resolutions of 2.43, 1.88, 3.01 and 2.65 for dansyl-Ser, dansyl-Met, dansyl-Thr and dansyl-Val, respectively. Furthermore, a comparison with microfluidic CZE was investigated demonstrating that muOTCEC was a promising method for rapid chiral recognition. PMID- 19137531 TI - Simultaneous separation of anionic, cationic, and neutral components in capillary liquid chromatography using mixed-bed column of hydrophilic and anion-exchange stationary phases. AB - A simultaneous separation of anionic, cationic, and neutral compounds was carried out in capillary LC using a mixed-bed column of zwitterionic hydrophilic and anion-exchange stationary phases. The combination in stationary phases was suitable to retain anionic, cationic, and neutral analytes. The retentions of anionic compounds were suppressed with the increase in the buffer concentration in the mobile phase whereas those for the cationic and neutral compounds were almost constant, i.e., the anionic analytes were mainly retained by anion exchange mode and the other compounds by the hydrophilic interaction. Both hydrophilic and electrostatic retentions were successfully controlled by the ACN content in the mobile phase. The gradient elution of increasing water content in the mobile phase achieved good separation for eight compounds found in metabolome analysis within 6 min. Both intra- (n = 7) and inter-day (n = 3) precisions (%RSD) of elution time for every eight compounds were less than 3 and 5%, respectively. PMID- 19137532 TI - Covalent modification of fused silica capillaries with quaternized polyamines to achieve robust and stable anodic electroosmotic flow. AB - Synthetic chemistry originally developed for the manufacture of chemically stable silica polyamine composites was adapted for the modification of fused silica capillaries for application in CE. Polyethyleneimine (PEI) and polyallylamine (PAA) were covalently bonded to the interior surface of fused silica capillaries utilizing 3-chloropropyltrichlorosilane (CPTCS) or 3-glycidoxypropyl trimethoxysilane (GPTMS) to anchor the polymers to the surface. The surface-bound polymers were subsequently quaternized using methyl iodide (MeI). The resulting modified capillaries were studied using CE, and were shown to provide reproducible, stable, and robust anodic EOF throughout the pH range of 2-10. Surface modifications utilizing CPTCS could be rinsed with up to 6 M HCL or 1 M NaOH without significant loss of surface modifier. The utility of the modified capillaries for the separation of simple anions and cations was demonstrated. PMID- 19137533 TI - Synthesis and in-vitro antimycobacterial evaluation of 1-(cyclopropyl/2,4 difluorophenyl/tert-butyl)-1,4-dihydro- 8-methyl-6-nitro-4-oxo-7-(substituted secondary amino)quinoline-3-carboxylic acids. AB - Fifty one newer 1-(cyclopropyl/2,4-difluorophenyl/tert-butyl)-1,4-dihydro-8 methyl-6-nitro-4-oxo-7-(substituted secondary amino)quinoline-3-carboxylic acids were synthesized from 1,3-dichloro-2-methylbenzene and evaluated for in-vitro antimycobacterial activities against Mycobacterium tuberculosis H37Rv (MTB), multi-drug resistant Mycobacterium tuberculosis (MDR-TB), and Mycobacterium smegmatis (MC(2)). Among the synthesized compounds, 1-cyclopropyl-1,4-dihydro-7 (3,4-dihydro-6,7-dimethoxyisoquinolin-2(1H)-yl)-8-methyl-6-nitro-4-oxoquinoline-3 carboxylic acid 9p was found to be the most active compound in vitro with a MIC value of 0.39 microM against MTB. Against MDR-TB, compound 7-(2-carboxy-5,6 dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)-1-cyclopropyl-1,4-dihydro-8-methyl-6-nitro 4-oxoquinoline-3-carboxylic acid 9n was found to be the most active with a MIC value of 0.09 microM. PMID- 19137534 TI - N-benzylsalicylthioamides: highly active potential antituberculotics. AB - A gseries of 29 new derivatives of N-benzylsalicylthioamides was synthesized and the compounds were tested for in-vitro antimycobacterial activity against Mycobacterium tuberculosis, Mycobacterium kansasii, and Mycobacterium avium. The activity was analyzed by quantitative structure-activity relationship (QSAR). Activity increased with increasing lipophilicity and electron donating effect of the substituents in the acyl moiety and decreased with the electrophilic superdelocalizability of the molecules. The most active compounds are more active than isoniazid (INH) and are active against INH-resistant potential pathogenic strains of mycobacterium. PMID- 19137535 TI - The protein environment drives selectivity for sulfide oxidation by an artificial metalloenzyme. AB - MAGIC Mn-salen mETALLOZYME: The design of an original, artificial, inorganic, complex-protein adduct, has led to a better understanding of the synergistic effects of both partners. The exclusive formation of sulfoxides by the hybrid biocatalyst, as opposed to sulfone in the case of the free inorganic complex, highlights the modulating role of the inorganic-complex-binding site in the protein. Artificial metalloenzymes based on the incorporation of Mn-salen complexes into human serum albumin display high efficiency and selectivity for sulfoxide production during sulfide oxidation. The reactions carried out by the artificial metallozymes are comparable to those carried out by natural biocatalysis. We have found that the polarity of the protein environment is crucial for selectivity and that a synergy between both partners of the hybrid results in the novel activity. PMID- 19137536 TI - Iron and ruthenium nanoparticles in carbon prepared by thermolysis of buckymetallocenes. AB - Thermolysis of fullerene iron and ruthenium complexes (buckymetallocene M(C(60)R(5))Cp (M = Fe; R = Ph (1) and Me (2), M = Ru; R = Ph (3), Me (4)) under a nitrogen atmosphere produced metal nanoparticles dispersed in carbon materials. The thermal degradation processes of the buckymetallocenes were studied by TG DTA, TEM with a heating sample stage, and VT-XRD. Variation of the thermolysis temperature led to a change in the size of the nanoparticles and the morphology of the carbon materials. Thermolysis of buckyferrocene at 700 degrees C gave highly dispersed iron nanoparticles (average diameter of 7.4 nm). After thermal treatment at 900 degrees C, graphite structures such as carbon nanocapsules and carbon nanotubes formed because of the catalytic activity of the iron nanoparticles. Ruthenium nanoparticles prepared from buckyruthenocene were much smaller than the iron counterparts, and did not catalyze the formation of graphite structures. When buckyruthenocene absorbed on silica gel was heated at 500 degrees C under a hydrogen atmosphere, the resulting ruthenium nanoparticles showed high activity in catalytic hydrogenation. PMID- 19137537 TI - Phenotype and function of neonatal DC. AB - Newborns face complex physical and immunological changes before and after birth. Although the uterus is a sterile environment for the fetus, it also contains non self material from the mother. Birth involves the transition from the sterile intra-uterine environment to an environment rich in microbes and requires rapid induction of appropriate responses to control these microbes. In this review we focus on the similarities and differences of human and murine neonatal DC and their reaction to various stimuli. A better understanding of the newborn immune system--in particular, the DC-T-cell interaction--will be beneficial for the development of improved strategies to prevent or treat infections in this vulnerable population and prepare the immune system to cope with allergens and tumors later in life. PMID- 19137538 TI - The role of the dental lamina in mammalian tooth replacement. AB - We have applied the ferret, Mustela putorius furo, as a model for tooth replacement. Ferret has a heterodont dentition, which includes all tooth families, and all antemolar teeth are replaced. Compared with mouse, the ferret therefore has a less derived mammalian dentition resembling that of humans. We have studied tooth replacement in serial histological sections in embryonic and young postnatal ferrets. Our observations indicate that the replacement teeth form from the dental lamina that is intimately connected to the lingual aspect of the deciduous tooth enamel organ. It grows as an offshoot from the enamel organ, elongates in cervical direction and later buds to give rise to the replacement tooth. The extent of the dental lamina growth, preceding replacement tooth budding, varied between different teeth. The dynamic gene expression patterns of Sostdc1, Shh and Axin2 brought new insight into the signal networks regulating the tooth replacement process. The distinct expression of Sostdc1 at the interface between the dental lamina and the deciduous tooth is the first indication of a specific tissue identity of the dental lamina. We suggest that the reactivation of a competent dental lamina is pivotal for the replacement tooth formation. PMID- 19137539 TI - Breast MR. PMID- 19137540 TI - Classification of cochlear implant failures and explantation: the Melbourne experience, 1982-2006. AB - The definition and categorization of reasons for cochlear implant (CI) failure have recently been standardized following the publication of the European Consensus Statement on Cochlear Implant Failures and Explantation (ECSCIFE) (2005). The aim of this study was to review the Melbourne experience with cochlear implant failure and reimplantation, applying ECSCIFE guidelines for categorization and assessing hearing outcomes. A retrospective review was carried out of Melbourne CI clinic's records to identify all cases of implant failure and categorize them using ECSCIFE guidelines. Comparison was made of pre- and post reimplantation hearing levels for those patients who had undergone ipsilateral reimplantation. Between September 1982 and October 2006 the Melbourne clinic conducted 1164 CIs with 62 implant failures and explantation (5.3%). Reasons included device failure, (ECSCIFE category C, n = 35, 3.0%), medical complication (ECSCIFE category D, n = 19), characteristics decrement (ECSCIFE category B1, n = 2) and performance decrement (ECSCIFE category B2 n = 6). Forty-nine patients underwent ipsilateral explantation/reimplantation. Auditory performance with the second implant was comparable to the first implant levels. The ECSCIFE provides an easy-to-use classification system for international reporting of CI device failures and explantations, however, detailed information of each implant failure may be lost in the final category, however. Cochlear explantation/reimplantation in Melbourne is a safe and effective procedure to restore patients to their pre operative best auditory function levels. PMID- 19137541 TI - Clusterin is a short half-life, poly-ubiquitinated protein, which controls the fate of prostate cancer cells. AB - The Clusterin (CLU) gene produces different forms of protein products, which vary in their biological properties and distribution within the cell. Both the extra- and intracellular CLU forms regulate cell proliferation and apoptosis. Dis regulation of CLU expression occurs in many cancer types, including prostate cancer. The role that CLU plays in tumorigenesis is still unclear. We found that CLU over-expression inhibited cell proliferation and induced apoptosis in prostate cancer cells. Here we show that depletion of CLU affects the growth of PC-3 prostate cancer cells. Following siRNA targeting all CLU mRNA variants, all protein products quickly disappeared, inducing cell cycle progression and higher expression of specific proliferation markers (i.e., H3 mRNA, PCNA, and cyclins A, B1, and D) as detected by RT-qPCR and Western blot. Quite surprisingly, we also found that the turnover of CLU protein is very rapid and tightly regulated by ubiquitin-proteasome mediated degradation. Inhibition of protein synthesis by cycloheximide showed that CLU half-life is less than 2 h. CLU protein products were found poly-ubiquitinated by co-immuniprecipitation. Proteasome inhibition by MG132 caused stabilization and accumulation of all CLU protein products, including the nuclear form of CLU (nCLU), and committing cells to caspase dependent death. In conclusion, proteasome inhibition may induce prostate cancer cell death through accumulation of nCLU, a potential tumor suppressor factor. PMID- 19137542 TI - Internalization of annexin A5-functionalized iron oxide particles by apoptotic Jurkat cells. AB - Apoptosis plays an important role in the etiology of various diseases. Several studies have reported on the use of annexin A5-functionalized iron oxide particles for the detection of apoptosis with MRI, both in vitro and in vivo. The protein annexin A5 binds with high affinity to the phospholipid phosphatidylserine, which is exposed in the outer leaflet of the apoptotic cell membrane. When co-exposed to apoptotic stimuli, this protein was shown to internalize into endocytic vesicles. Therefore in the present study we investigated the possible internalization of commercially available annexin A5 functionalized iron oxide particles (r1 = 34.0 +/- 2.1 and r2 = 205.0 +/- 10.4 mm(-1) s(-1) at 20 MHz), and the effects of their spatial distribution on relaxation rates R2*, R2 and R1. Two different incubation procedures were performed, where (1) Jurkat cells were either incubated with the contrast agent after induction of apoptosis or (2) Jurkat cells were simultaneously incubated with the apoptotic stimulus and the contrast agent. Transmission electron microscopy images and relaxation rates showed that the first incubation strategy mainly resulted in binding of the annexin A5-iron oxide particles to the cell membrane, whereas the second procedure allowed extensive membrane-association as well as a small amount of internalization. Owing to the small extent of internalization, only minor differences were observed between the DeltaR2*/DeltaR2 and DeltaR2/DeltaR1 ratios of cell pellets with membrane associated or internalized annexin A5 particles. Only the increase in R1 (DeltaR1) appeared to be diminished by the internalization. Internalization of annexin A5-iron oxide particles is also expected to occur in vivo, where the apoptotic stimulus and the contrast agent are simultaneously present. Where the extent of internalization in vivo is similar to that observed in the present study, both T2- and T2*-weighted MR sequences are considered suitable for the detection of these particles in vivo. PMID- 19137543 TI - Unintended molecular interactions in transgenic plants expressing clinically useful proteins: the case of bovine aprotinin traveling the potato leaf cell secretory pathway. AB - We assessed the impact of subcellular targeting on the heterologous expression of a clinically useful protease inhibitor, bovine aprotinin, in leaves of potato, Solanum tuberosum. Transgenic potato lines targeting aprotinin to the cytosol, the ER or the apoplast were first generated, and then assessed for their ability to accumulate the recombinant protein. On-chip detection and quantitation of aprotinin variants by SELDI TOF MS showed the inhibitor to be absent in the cytosol, but present under different forms in the ER and the apoplast. No visible phenotypic effects of aprotinin were observed for the transgenic lines, but aprotinin retention in the ER was associated with a significant decrease of leaf soluble protein content. A 2-D gel assessment of control and transgenic lines revealed a possible link between this altered protein content and the down regulation of proteins implicated in protein synthesis and maturation. These observations, supported by complementary 2-DE analyses with potato lines targeting aprotinin to the apoplast, suggest an aprotinin-mediated feedback in planta negatively altering protein anabolism. From a practical viewpoint, these data illustrate the importance of taking into account not only the characteristics of recombinant proteins expressed in heterologous environments, but also their possible effects on protein accumulation in the host plant factory. PMID- 19137544 TI - Highly sensitive saturation labeling reveals changes in abundance of cell cycle associated proteins and redox enzyme variants during oocyte maturation in vitro. AB - Oocyte maturation is a complex process and a critical issue in assisted reproduction techniques (ART) in humans and other mammals. We used a sensitive 2 D DIGE saturation labeling approach including an internal pooled standard for quantitative proteome profiling of immature versus in vitro matured bovine oocytes in six independent samples. The study comprised 48 2D gel images representing 24 DIGE experiments. From 250 ng sample analyzed per gel, quantitative analysis revealed an average of 2244 spots in pH 4-7 images and 1291 spots in pH 6-9 images. Thirty-eight spots with different intensities were detected in total. Spots of a preparative gel from 2200 oocytes were identified by nano-LC-MS/MS analysis. The ten spots which could be unambiguously identified include the Ca2+-binding protein translationally controlled tumor protein, enzymes of the Krebs and pentose phosphate cycles, clusterin, 14-3-3 epsilon, elongation factor-1 gamma, and redox enzymes such as polymorphic forms of GST Mu 5 and peroxiredoxin-3. The cellular distribution of two proteins was determined by confocal laser scanning microscopy. The interesting protein candidates identified by this study may help to improve the in vitro maturation process in order to increase the rate of successful in vitro fertilization and other ART in cattle and other mammals. PMID- 19137545 TI - Dissimilar hepatic protein expression profiles during the acute and flow phases following experimental thermal injury. AB - The liver plays a major role in the early hypometabolic and later hypermetabolic phases after severe burn injury. Proteomic analysis was used to identify altered proteins in liver during these two phases. Sprague-Dawley rats were subjected to a full-thickness dorsal burn injury covering 40% of the total body surface area. Controls consisted of sham-treated animals. Liver tissues were collected on postburn days 1 and 7. The proteomic data show greater production of positive acute phase proteins on day 1 than on day 7. Many antioxidant enzymes were coordinately downregulated on day 1, including the potent biliverdin reductase. These antioxidants were restored and in some cases upregulated on day 7. This opposite trend in the change of antioxidant proteins corroborated our finding of more pronounced oxidative stress on day 1 than on day 7 as measured via protein carbonyl content. The changes of metabolic enzymes on days 1 and 7 were consistent with hypo- and hyper-metabolic states, respectively. Furthermore, a previously unreported decrease in ornithine aminotransferase on day 7 may be a key contributor to the observed increased urinary urea excretion during the hypermetabolic phase. Overall, the many differences in protein expression observed on postburn days 1 and 7 reflect the dissimilar hepatic metabolic patterns during the acute and flow phases following burn injury. PMID- 19137546 TI - A comprehensive evaluation of imidazole-zinc reverse stain for current proteomic researches. AB - In this paper, we comprehensively evaluated the capability of imidazole-zinc reverse stain (ZN) in comparative proteomics. Three commonly used protein gel staining methods, including silver (SN), SYPRO Ruby (SR), and CB stain were investigated alongside for comparison purpose. A transparency scanning procedure, which may deliver more even and contrasting gel images, was found best for documenting ZN stained gels. Our results showed that ZN was more sensitive than SN, SR, and CB. It may reveal as few as 1.8 ng of proteins in a gel. Moreover, ZN was found to provide a linear dynamic range of staining for revealing proteins up to 140 ng, and show an insignificant staining preference. To analyze a ZN stained 2-D gel image that generally comprises an apparent but even background, the Melanie 4 software was found more suitable than others. Furthermore, ZN demonstrated an equivalent or better MS compatibility than the other three staining methods. Intense and comprehensive MS profiles were frequently observed for ZN stained gel spots. Approximate two-third of ZN stained gel spots were successfully identified for protein identities. Taken together, our results suggest that the prompt, cost effective and versatile ZN is well suited for current proteomic researches. PMID- 19137547 TI - The proteome of Salmonella Typhimurium grown under in vivo-mimicking conditions. AB - To successfully infect a host, it is a prerequisite for enteric pathogens such as Salmonella enterica serovar Typhimurium to adapt to their environment, in casu the gastrointestinal tract. The adoption of an appropriate lifestyle is triggered by environmental signals such as the low oxygen availability and high osmolarity prevalent in the gut. In order to gain more insight in the changes that are induced when S. Typhimurium is adapting to these particular conditions, we used 2 D DIGE technology to investigate the combined effect of low oxygen tension and high osmolarity on the proteome of S. Typhimurium SL1344 compared to standard laboratory conditions. As a validation of the 2-D DIGE technique, preferential protein labeling by the Cy-dyes was assessed and proved to be negligible. The differentially expressed proteins identified reflect very well the applied culture conditions. Furthermore, reported transcriptional changes and observed changes at the translational level show overlap. Among the metabolic processes that are upregulated under in vivo-mimicking conditions are anaerobic fumarate respiration and the utilization of 1,2-propanediol. We also provide evidence that S. Typhimurium expresses an arginine deiminase pathway for the catabolism of L arginine. The increased activity of this pathway was biochemically validated. Finally, also proteins involved in quorum sensing and virulence are differentially expressed under in vivo-mimicking conditions. These conditions offer possibilities as a simplified model system for the host environment given the high overlap of identifications in our study and reported genuine in vivo studies, respectively. PMID- 19137548 TI - Yeast protein expression profile during acetic acid-induced apoptosis indicates causal involvement of the TOR pathway. AB - Although acetic acid has been shown to induce apoptosis in yeast, the exact apoptotic mechanisms remain unknown. Here, we studied the effects of acetic acid treatment on yeast cells by 2-DE, revealing alterations in the levels of proteins directly or indirectly linked with the target of rapamycin (TOR) pathway: amino acid biosynthesis, transcription/translation machinery, carbohydrate metabolism, nucleotide biosynthesis, stress response, protein turnover and cell cycle. The increased levels of proteins involved in amino-acid biosynthesis presented a counteracting response to a severe intracellular amino-acid starvation induced by acetic acid. Deletion of GCN4 and GCN2 encoding key players of general amino-acid control (GAAC) system caused a higher resistance to acetic acid indicating an involvement of Gcn4p/Gcn2p in the apoptotic signaling. Involvement of the TOR pathway in acetic acid-induced apoptosis was also reflected by the higher survival rates associated to a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-negative phenotype and lower reactive oxygen species levels of Deltator1 cells. In addition, deletion mutants for several downstream mediators of the TOR pathway revealed that apoptotic signaling involves the phosphatases Pph21p and Pph22p but not Sit4p. Altogether, our results indicate that GAAC and TOR pathways (Tor1p) are involved in the signaling of acetic acid-induced apoptosis. PMID- 19137549 TI - A chip-based amide-HILIC LC/MS platform for glycosaminoglycan glycomics profiling. AB - A key challenge to investigations into the functional roles of glycosaminoglycans (GAGs) in biological systems is the difficulty in achieving sensitive, stable, and reproducible mass spectrometric analysis. GAGs are linear carbohydrates with domains that vary in the extent of sulfation, acetylation, and uronic acid epimerization. It is of particular importance to determine spatial and temporal variations of GAG domain structures in biological tissues. In order to analyze GAGs from tissue, it is useful to couple MS with an on-line separation system. The purposes of the separation system are both to remove components that inhibit GAG ionization and to enable the analysis of very complex mixtures. This contribution presents amide-silica hydrophilic interaction chromatography (HILIC) in a chip-based format for LC/MS of heparin, heparan sulfate (HS) GAGs. The chip interface yields robust performance in the negative ion mode that is essential for GAGs and other acidic glycan classes while the built-in trapping cartridge reduces background from the biological tissue matrix. The HILIC chromatographic separation is based on a combination of the glycan chain lengths and the numbers of hydrophobic acetate (Ac) groups and acidic sulfate groups. In summary, chip based amide-HILIC LC/MS is an enabling technology for GAG glycomics profiling. PMID- 19137551 TI - Identification of secreted proteins regulated by cAMP in glioblastoma cells using glycopeptide capture and label-free quantification. AB - Exposure of glioblastoma U87MG cells to a cAMP analog leads to a decrease in proliferation, invasion, and angiogenic potential. Here, we apply a label-free MS based approach to identify formerly N-linked glycopeptides that change in abundance upon cAMP treatment. Over 150 unique glycopeptides in three biological repetitions were quantified, leading to the identification of 14 upregulated proteins and 21 downregulated proteins due to cAMP treatment. Of these, eight have been validated, either through comparison with microarray data or by Western blot. We estimate our ability to identify differentially expressed peptides at greater than 85% in a single biological repetition, while the analysis of multiple biological repetitions lowers the false positive rate to approximately 2%. Many of the proteins identified in this study are involved in cell signaling and some, such as Tenascin C, Cathepsin L, Neuroblastoma suppressor of tumorigenicity, and AXL/UFO tyrosine-protein kinase receptor, have been previously shown to be involved in glioblastoma progression. We also identify several semitryptic peptides that increase in abundance upon cAMP treatment, suggesting that cAMP regulates protease activity in these cells. Overall, these results demonstrate the benefits of using a highly specific enrichment method for quantitative proteomic experiments. PMID- 19137550 TI - Effects of green tea extract on lung cancer A549 cells: proteomic identification of proteins associated with cell migration. AB - Green tea polyphenols exhibit multiple antitumor activities, and the mechanisms of action are not completely understood. Previously, we reported that green tea extract (GTE)-induced actin remolding is associated with increased cell adhesion and decreased motility in A549 lung cancer cells. To identify the cellular targets responsible for green tea-induced actin remodeling, we performed 2-DE LC MS/MS of A549 cells before and after GTE exposure. We have identified 14 protein spots that changed in expression (> or =2-fold) after GTE treatment. These proteins are involved in calcium-binding, cytoskeleton and motility, metabolism, detoxification, or gene regulation. In particular we found upregulation of several genes that modulate actin remodeling and cell migration, including lamin A/C. Our data indicated that GTE-induced lamin A/C upregulation appears to be at the transcriptional level and the increased expression results in the decrease in cell motility, as confirmed by siRNA. The result of the study demonstrates that GTE alters the levels of many proteins involved in growth, motility and apoptosis of A549 cells and their identification may explain the multiple antitumor activities of GTE. PMID- 19137552 TI - Multivariate analysis of single quadrupole LC-MS spectra for routine characterization and quantification of intact proteins. AB - Modern high-throughput proteomic platforms allow incomparable protein mixture resolution and identification. However, such sophisticated facilities are expensive and not always accessible for routine analysis of simple mixtures. In this paper, we propose a simple methodology, based on detection of intact, nondigested proteins by LC coupled to single quadrupole MS (sqLC-MS), followed by the analysis of the resulting spectra by multivariate analysis (MA). By doing so, even large molecular weight (MW) proteins, generating complex spectra, can be characterized to a level that allows isoform discrimination, while standard algorithms, such as MS spectrum deconvolution, cannot. To demonstrate the effectiveness of the proposed approach, we have analyzed the spectra of a set of purified, intact albumins from seven different organisms (bovine, human, rabbit, rat, sheep, mouse, and pig) as a model of microheterogenous proteins, using Projection to Latent Structure Discriminant Analysis (PLS-DA). Although these proteins are very similar (less than 1% difference in MW), sqLC-MS/MA allowed their classification, and the identification of unknown source samples. In addition, MA allowed precise protein quantification from the same data (calibration curve R2 = 0.9966). The ability to rapidly characterize and quantify proteins, together with simplicity and affordability, could make of combined sqLC MS/MA a routine method for the characterization of simple mixture of known proteins. PMID- 19137553 TI - Mass spectrometric characterization of membrane integral low molecular weight proteins from photosystem II in barley etioplasts. AB - In Photosystem II (PSII), a high number of plastid encoded and membrane integral low molecular weight proteins smaller than 10 kDa, the proteins PsbE, F, H, I, J, K, L, M, N, Tc, Z and the nuclear encoded PsbW, X, Y1, Y2 proteins have been described. Here we show that all low molecular weight proteins of PSII already accumulate in the etioplast membrane fraction in darkness, whereas PsaI and PsaJ of photosystem I (PSI) represent the only low molecular weight proteins that do not accumulate in darkness. We found by BN-PAGE separation of membrane protein complexes and selective MS that the accumulation of one-helix proteins from PSII is light independent and occurs in etioplasts. In contrast, in chloroplasts isolated from light-grown plants, low molecular weight proteins were found to specifically accumulate in PSI and II complexes. Our results demonstrate how plants grown in darkness prepare for the induction of chlorophyll dependent photosystem assembly upon light perception. We anticipate that our investigation will provide the essential means for the analysis of protein assembly in any membrane utilizing low molecular weight protein subunits. PMID- 19137554 TI - Insights into yeast adaptive response to the agricultural fungicide mancozeb: a toxicoproteomics approach. AB - Toxicogenomics has the potential to elucidate gene-environment interactions to identify genes that are affected by a particular chemical at the early stages of the toxicological response and to establish parallelisms between different organisms. The fungicide mancozeb, widely used in agriculture, is an ethylene-bis dithiocarbamate complex with manganese and zinc. Exposure to this pesticide has been linked to the development of idiopathic Parkinson's disease and cancer. Given that many signalling pathways and their molecular components are substantially conserved among eukaryotic organisms, we used Saccharomyces cerevisiae to get insights into the molecular mechanisms of mancozeb toxicity and adaptation based on expression proteomics. The early global response to mancozeb was analysed by quantitative proteomics using 2-DE. The target genes (e.g. TSA1, TSA2, SOD1, SOD2, AHP1, GRE2, GRX1, CYS3, PRE3, PRE6, PRE8, PRE9, EFT1, RPS5, TIF11, HSP31, HSP26, HSP104, HSP60, HSP70-family) and the putative main transcription activators (e.g. Yap1, Msn2/Msn4, Met4, Hsf1, Aft1, Pdr1, Skn7, Rpn4p, Gcn4) of the complex mancozeb-induced expression changes are related with yeast response to stress, in particular to oxidative stress, protein translation initiation and protein folding, disassembling of protein aggregates and degradation of damaged proteins. Our results also suggest that this study provided powerful indications that may be useful to expand the knowledge obtained in yeast not only to the global response to mancozeb toxicity in phytopathogenic fungi but also to humans. PMID- 19137555 TI - Improved protein sequence coverage by on resin deglycosylation and cysteine modification for biomarker discovery. AB - Membrane proteins and secreted factors (soluble proteins or extracellular matrix components) are the targets of most monoclonal antibodies, which are currently in clinical development. These proteins are frequently post-translationally modified, e.g. by the formation of disulfide bonds or by glycosylation, which complicates their identification using proteomics technologies. Here, we describe a novel methodology for the on resin deglycosylation and cysteine modification of proteins after in vitro, in vivo or ex vivo biotinylation. Biotinylated proteins are captured on streptavidin resin and all subsequent modifications, as well as the proteolytic digestion, which yields peptides for MS analysis, are performed on resin. Using biotinylated bovine fetuin-A as a test protein, an improvement in sequence coverage from 7.9 to 58.7% could be shown, including the identification of all three glycosylation sites. Furthermore, a complex mixture derived from the ex vivo biotinylation of vascular structures in human kidney with cancer obtained by perfusion after surgical resection revealed almost a doubling of sequence coverage for all checked proteins when analyzed by LC-MALDI TOF/TOF. PMID- 19137556 TI - Analysis of the subproteomes of proteinases and heparin-binding toxins of eight Bothrops venoms. AB - Viperid snakes show the most complex snake-venom proteomes and offer an intriguing challenge in terms of understanding the nature of their components and the pathological outcomes of envenomation characterized by local and systemic effects. In this work, the venom complexity of eight Bothrops species was analyzed by 2-DE, and their subproteomes of proteinases were explored by 2-D immunostaining and 2-D gelatin zymography, demonstrating the diversity of their profiles. Heparin, a highly sulfated glycosaminoglycan released from mast cells, is involved in anti-coagulant and anti-inflammatory processes. Here, we explored the hypothesis that heparin released upon envenomation could interact with toxins and interfere with venom pathogenesis. We first identified the Bothrops venom subproteome of toxins that bind with high-affinity for heparin as composed of mainly serine proteinases and C-type lectins. Next, we explored the Bothrops jararaca toxins that bind to heparin under physiological conditions and identified a relationship between the subproteomes of proteinases, and that of heparin-binding toxins. Only the non-bound fraction, composed mainly of metalloproteinases, showed lethal and hemorrhagic activities, whereas the heparin bound fraction contained mainly serine proteinases associated with coagulant and fibrinogenolytic activities. These data suggest that heparin binding to B. jararaca venom components in vivo has a minor protective effect to venom toxicity. PMID- 19137557 TI - Analysis of outer membrane porin complexes of Neisseria meningitidis in wild-type and specific knock-out mutant strains. AB - The structure of the porin complexes of Neisseria meningitidis was assessed in the vaccine strain H44/76 and its homologous mutants lacking the main porins (PorA and PorB) and other outer membrane (OM) components (RmpM and FetA). The analysis using 1-D blue native (BN) electrophoresis, 2-D BN/SDS-PAGE and 2-D diagonal electrophoresis, followed by LC/MS-MS (for 1-D gels) or MALDI-TOF (for 2 D gels) revealed at least six porin complexes in the wild-type strain with molecular masses (MW) ranging from 145 to 195 kDa and variable composition: The two higher MW complexes are formed by PorA, PorB and RmpM, the following three are formed by PorA and PorB, and the lower MW one is formed by only PorB. Complexes in the mutants lacking either PorA, PorB or RmpM, but not those in the mutant lacking FetA, were alterered respect to those in the wild-type strain. The most evident alteration was seen in the mutant lacking PorB, in which PorA formed only a high MW complex (approximately 800 kDa). Our results suggest that PorA and PorB could form a 'basic' template for the transportation systems in the OM of the meningococci. Other proteins (such as RmpM) could be transiently associated to the porin complexes, depending on the specific tranport needs at different stages of the meningococcal life cycle, resulting in a dynamic net of pores of variable composition. PMID- 19137558 TI - The neuropeptidome of Rhodnius prolixus brain. AB - We show a sensitive and straightforward off-line nano-LC-MALDI-MS/MS workflow that allowed the first comprehensive neuropeptidomic analysis of an insect disease vector. This approach was applied to identify neuropeptides in the brain of Rhodnius prolixus, a vector of Chagas disease. This work will contribute to the annotation of genes in the ongoing R. prolixus genome sequence project. Peptides were identified by de novo sequencing and comparisons to known neuropeptides from different organisms by database search. By these means, we were able to identify 42 novel neuropeptides from R. prolixus. The peptides were classified as extended FMRF-amide-related peptides, sulfakinins, myosuppressins, short neuropeptide F, long neuropeptide F, SIF-amide-related peptides, tachykinins, orcokinins, allatostatins, allatotropins, calcitonin-like diuretic hormones, corazonin, and pyrokinin. Some of them were detected in multiple isoforms and/or truncated fragments. Interestingly, some of the R. prolixus peptides, as myosuppressin and sulfakinins, are unique in their characteristic C terminal domain among insect neuropeptides identified so far. PMID- 19137560 TI - Highly selective dehydrogenative silylation of alkenes catalyzed by rhenium complexes. AB - Choosy chemicals: Rhenium(I) complexes of type [ReBr(2)(L)(NO)(PR(3))(2)] (L=H(2) (1), CH(3)CN (2), ethylene (3); R=iPr (a), cyclohexyl (b)) proved to be suitable catalyst precursors for the highly selective dehydrogenative silylation of alkenes. Two types of rhenium(I) hydride species, [ReBrH(NO)(PR(3))(2)] (4) and [ReBr(eta(2)-CH(2)=CHR(1))H(NO)(PR(3))(2)] (5), were found in the [ReBr(2)(L)(NO)(PR(3))(2)]-catalyzed dehydrogenative silylation of alkenes.Rhenium(I) complexes of type [ReBr(2)(L)(NO)(PR(3))(2)] (L=H(2) (1), CH(3)CN (2), and ethylene (3); R=iPr (a) and cyclohexyl (Cy; b)) catalyze dehydrogenative silylation of alkenes in a highly selective manner to yield silyl alkenes and the corresponding alkanes. Hydrosilylation products appear only rarely depending on the type of olefinic substituent, and if they do appear then it is in very minor amounts. Mechanistic studies showed that two rhenium(I) hydride species of type [ReBrH(NO)(PR(3))(2)] (R=iPr (4 a) and Cy (4 b)) and [ReBr(eta(2)-CH(2)=CHR(1))H(NO)(PR(3))(2)] (R(1)=p-CH(3)C(6)H(4), R=iPr (5 a), Cy (5 b); R(1)=H, R=iPr (5 a'), Cy (5 b')) are involved in the initiation pathway of the catalysis. The rate-determining steps of the catalytic cycle are the phosphine dissociation from complexes of type 5 and the reductive eliminations to form the alkane components. The catalytic cycle implies that the given rhenium systems have the ability to activate C-H and Si-H bonds through the aid of a facile redox interplay of Re(I) and Re(III) species. The molecular structures of 4 b and 5 a were established by means of X-ray diffraction studies. PMID- 19137559 TI - Base-pairing directed folding of a bimolecular G-quadruplex: new insights into G quadruplex-based DNAzymes. AB - Base pairs, magic hands: An additional base-pairing duplex is utilized to control the folding topologies of a bimolecular G-quadruplex formed by two G-rich single stranded DNAs (see picture), which is dependent on the position of base pairs. This study clearly reveals an important intrinsic role of additional base pairs in the G-quadruplex structure, and also provides a clue to the formation mechanism of the G-quadruplex-based DNAzyme. PMID- 19137561 TI - Selective chromofluorogenic sensing of heparin by using functionalised silica nanoparticles containing binding sites and a signalling reporter. AB - Heparin detective: Silica nanoparticles functionalised with ion-channel scaffolds were prepared and used for the chromofluorogenic sensing of heparin in aqueous environments (see figure). The surface of the nanoparticles was functionalised with polyamines (binding sites) and thiols. The reaction of a dye (squaraine) with the surface thiol groups was selectively inhibited by the coordination of heparin with the partly protonated polyamines. PMID- 19137563 TI - Ultrafast photoinduced processes in alizarin-sensitized metal oxide mesoporous films. AB - Close to the edge: Photoexcitation of alizarin coupled to the surface of mesoporous TiO(2) films leads to ultrafast electron transfer to the TiO(2) conduction band (see picture). Complex kinetics after photoexcitation depend on the excitation energy, and indicate a position of the alizarin excited state close to the TiO(2) conduction band edge, where the density of acceptor states is reduced. The photoinduced dynamics in Al(2)O(3) and TiO(2) mesoporous films sensitized by the strongly coupled alizarin dye is investigated by femtosecond transient absorption spectroscopy in the spectral range from UV to mid-IR. Alizarin/Al(2)O(3) acts as a nonreactive reference system, in which no electron transfer is observed. For comparison, the photoexcitation of the alizarin dye coupled to the surface of TiO(2) films leads to ultrafast electron transfer from the dye to the TiO(2) conduction band on the sub-100-fs timescale. We observe a fast relaxation of the alizarin excited state as well as a fast recombination of injected electrons with the alizarin cation on the picosecond timescale, which gives rise to very complex kinetics at short delay times. The infrared measurements clearly indicate that trapping of injected electrons is the main mechanism responsible for the observed long-lived charge separation in TiO(2) mesoporous films. The experimental findings can be explained by a position of the dye excited state close to the conduction band edge. PMID- 19137564 TI - Ab initio study of the interactions between CO(2) and N-containing organic heterocycles. AB - In the garden of dispersion: High-accuracy ab initio calculations are performed to determine the nature of the interactions and the most favorable geometries between CO(2) and heteroaromatic molecules containing nitrogen (see figure). Dispersion forces play a key role in the stabilization of the dimer, because correlation effects represent about 50 % of the total interaction energy. The interactions between carbon dioxide and organic heterocyclic molecules containing nitrogen are studied by using high-accuracy ab initio methods. Various adsorption positions are examined for pyridine. The preferred configuration is an in-plane configuration. An electron donor-electron acceptor (EDA) mechanism between the carbon of CO(2) and the nitrogen of the heterocycle and weak hydrogen bonds stabilize the complex, with important contributions from dispersion and induction forces. Quantitative results of the binding energy of CO(2) to pyridine (C(5)H(5)N), pyrimidine, pyridazine, and pyrazine (C(4)H(4)N(2)), triazine (C(3)H(3)N(3)), imidazole (C(3)H(4)N(2)), tetrazole (CH(2)N(4)), purine (C(5)H(4)N(4)), imidazopyridine (C(6)H(5)N(3)), adenine (C(5)H(5)N(5)), and imidazopyridamine (C(6)H(6)N(4)) for the in-plane configuration are presented. For purine, three different binding sites are examined. An approximate coupled cluster model including single and double excitations with a perturbative estimation of triple excitations (CCSD(T)) is used for benchmark calculations. The CCSD(T) basis-set limit is approximated from explicitly correlated second order Moller-Plesset (MP2-F12) calculations in the aug-cc-pVTZ basis in conjunction with contributions from single, double, and triple excitations calculated at the CCSD(T)/6-311++G** level of theory. Extrapolations to the MP2 basis-set limit coincide with the MP2-F12 calculations. The results are interpreted in terms of electrostatic potential maps and electron density redistribution plots. The effectiveness of density functional theory with the empirical dispersion correction of Grimme (DFT-D) is also examined. PMID- 19137565 TI - Nanostructured vanadium oxide model catalysts for selective oxidation reactions. AB - Controlled synthesis based on spectroscopic characterization, structure, and catalytic performance of mesoporous silica SBA-15-supported vanadium oxide model catalysts (see TEM image) are reviewed. The effect of water on the structure and dispersion of highly dispersed vanadium oxide is discussed in the light of recent results in multiple in situ spectroscopy.Characterization of the synthesis, structure, and catalytic performance of vanadium oxide catalysts supported on mesoporous silica SBA-15 for selective oxidation reactions are reviewed. Controlled synthesis on the basis of surface functionalization and ion exchange allows homogeneous deposition of vanadium oxide within the pores of SBA-15. Catalytic tests on the selective oxidation of methanol and propane demonstrate the full catalytic function of these model powder catalysts. Characterization of the dehydrated state by vibrational spectroscopy provides insight into the structure of SBA-15-supported vanadium oxide at low loadings. The effect of water on the structure and dispersion of highly dispersed vanadium oxide is discussed in light of recent results from multiple in situ spectroscopy. PMID- 19137567 TI - The molecular cat. AB - A manifestation of electronic entanglement in core-level spectroscopic measurements of diatomic molecules, reported recently by Schoffler and co workers, is discussed. The results are reminiscent of Schrodinger's famous Gedanken experiment with the cat (see picture). PMID- 19137566 TI - Preparation of highly luminescent CdTe/CdS core/shell quantum dots. AB - An effective shell-coating route is developed for covering oil-soluble CdTe quantum dots (QDs), which usually tend to aggregate during the heating-up process involved in shell formation. The new route is based on balancing the coordinating capacity and the activation effect of selected surfactants. The thus obtained highly luminescent CdTe/CdS core/shell QDs exhibit photoluminescence quantum yields as high as 92%--among the best results obtained so far for luminescent semiconductor nanocrystals. By changing the size of the CdTe core, or the thickness of the CdS shell, the emission colors of the obtained core/shell nanocrystals can be tuned between the visible and near-infrared regions of the spectrum following an identical procedure. PMID- 19137568 TI - Stoichiometric analysis of dissolved organic carbon flux into storage and growth in aerobic granules culture. AB - This study attempted to address a fundamental question of whether metabolic behaviors of aerobic granules are different from their counterparts, such as activated sludge and biofilms. A series of respirometric experiments were carried out using mature aerobic granules with mean sizes of 0.75-3.4 mm. Results suggested that metabolism of aerobic granules comprised three consecutive phases: (i) conversion of external dissolved organic carbon to a poly-beta hydroxybutyrate-like substance; (ii) growth of aerobic granules on the stored poly-beta-hydroxybutyrate-like substance derived from phase I, and (iii) subsequent endogenous metabolism of aerobic granules. The stoichiometric analysis revealed that the conversion yields of external dissolved organic carbon to the poly-beta-hydroxybutyrate-like substance, the growth yields of biomass on storage, and the overall growth yields of biomass on external dissolved organic carbon were not significantly correlated to the sizes of aerobic granules, i.e., the metabolism of aerobic granules would be size independent. The conversion coefficients and growth rates of aerobic granules were found to be comparable with those reported in the activated sludge and biofilms cultures, indicating that there would not be significant difference in the metabolisms of aerobic granules over activated sludge and biofilms. This information will be useful for modeling and designing aerobic granular sludge processes. PMID- 19137569 TI - Identification of microdeletions in candidate genes for cleft lip and/or palate. AB - BACKGROUND: Genome-wide association studies are now used routinely to identify genes implicated in complex traits. The panels used for such analyses can detect single nucleotide polymorphisms and copy number variants, both of which may help to identify small deleted regions of the genome that may contribute to a particular disease. METHODS: We performed a candidate gene analysis involving 1,221 SNPs in 333 candidate genes for orofacial clefting, using 2,823 samples from 725 two- and three-generation families with a proband having cleft lip with or without cleft palate. We used SNP genotyping, DNA sequencing, high-resolution DNA microarray analysis, and long-range PCR to confirm and characterize the deletion events. RESULTS: This dataset had a high duplicate reproducibility rate (99.98%), high Mendelian consistency rate (99.93%), and low missing data rate (0.55%), which provided a powerful opportunity for deletion detection. Apparent Mendelian inconsistencies between parents and children suggested deletion events in 15 individuals in 11 genomic regions. We confirmed deletions involving CYP1B1, FGF10, SP8, SUMO1, TBX1, TFAP2A, and UGT7A1, including both de novo and familial cases. Deletions of SUMO1, TBX1, and TFAP2A are likely to be etiologic. CONCLUSIONS: These deletions suggest the potential roles of genes or regulatory elements contained within deleted regions in the etiology of clefting. Our analysis took advantage of genotypes from a candidate-gene-based SNP survey and proved to be an efficient analytical approach to interrogate genes potentially involved in clefting. This can serve as a model to find genes playing a role in complex traits in general. PMID- 19137570 TI - Severe neurologic side effects in patients being treated for hemophagocytic lymphohistiocytosis. AB - BACKGROUND: Hemophagocytic Lymphohistiocytosis (HLH) is characterized by uncontrolled inflammation that is generally fatal without immune modulating chemotherapy. At Texas Children's Hospital, we have observed significant central nervous system (CNS) toxicity in several patients treated for HLH according to the Histiocyte Society protocol HLH-2004 in which cyclosporine is given early in the treatment regimen. METHODS: Patients diagnosed with HLH at Texas Children's Hospital between April 2004 and October 2007 were identified and charts were reviewed. A reference group of patients treated between August 2001 and March 2004, prior to the introduction of HLH-2004, was also evaluated. RESULTS: Five of 17 patients in the study group developed severe neurotoxicity. Four had new onset seizures associated with significant MRI abnormalities, while the fifth died of intracerebral hemorrhage. Timing of the development of neurologic side effects ranged from day 5 to week 6 of therapy. Cyclosporine levels were outside the therapeutic range (200-300 ng/ml) prior to the onset of symptoms in two of the five patients. Systolic blood pressures for all five patients were greater than the 95th percentile for age on at least one measurement within 24 hr of the onset of neurologic symptoms. MRI scans obtained within 24 hr of seizure activity in four patients were consistent with posterior reversible encephalopathy syndrome (PRES). By comparison only one patient in the reference group (n = 15) had neurotoxicity (PRES). CONCLUSIONS: Patients being treated for HLH appear to be at risk for neurotoxicity, particularly PRES. Elevated blood pressure, worsening renal and liver function, increased cyclosporine levels, and CNS involvement of HLH may be triggers for the neurotoxic side effects of treatment. Patients being treated on HLH-2004 require close monitoring of their neurologic status and modifiable risk factors such as hypertension should managed aggressively. If larger studies validate our observations, it will be important to determine if up front cyclosporine in HLH protocols confers a survival benefit that outweighs the potential risk of increased neurotoxicity. PMID- 19137571 TI - Prognostic index for patients with parotid carcinoma: international external validation in a Belgian-German database. AB - BACKGROUND: Prognostic indices for recurrence-free interval in patients with parotid carcinoma were developed and validated in a nationwide database. International validation would increase generalizability. METHODS: In a Belgian German database that contained 237 consecutive patients with parotid carcinoma, a pretreatment prognostic index (PS1) and a post-treatment prognostic index (PS2) were validated by calculating both indices for each patient, comparing coefficients, constructing survival curves, calculating the concordance measure C, and performing Wald tests for scale and weight optimization of included variables and for the possible inclusion of new variables. RESULTS: Sixty-nine percent of patients (standard error, 5%) were disease free at 5 years. The defined cutoff points for PS1 resulted in 5-year disease-free rates from 94% (PS1 = 1) to 42% (PS1 = 4), and the cutoff points for PS2 resulted in 5-year disease free rates from 93% (PS2 = 1) to 40% (PS2 = 4). Concordance measure C was good with 0.74 for both PS1 and PS2. Neither index could be improved statistically using this international database. There was some evidence that additional inclusion of the variable 'number of positive lymph nodes in the neck dissection specimen' could enhance the prognostic power of PS2. CONCLUSIONS: The prognostic indices performed adequately in this validation sample. Prospective generalized use seems to be well supported. PMID- 19137572 TI - Neonatal separation stress reduces glial fibrillary acidic protein- and S100beta immunoreactive astrocytes in the rat medial precentral cortex. AB - The interactions between the mother/parents and their offspring provides socioemotional input, which is essential for the establishment and maintenance of synaptic networks in prefrontal and limbic brain regions. Since glial cells are known to play an important role in developmental and experience-driven synaptic plasticity, the effect of an early adverse emotional experience induced by maternal separation for 1 or 6 h on the expression of the glia specific proteins S100beta and glial fibrillary acidic protein (GFAP) was quantitatively analyzed in anterior cingulate cortex, hippocampus, and precentral medial cortex. Three animal groups were analyzed at postnatal day 14: (i) separated for 1 h; (ii) separated for 6 h; (iii) undisturbed (control). Twenty-four hours after stress exposure, the stressed brains showed significantly reduced numbers of S100beta immunoreactive (ir) cells in the anterior cingulate cortex (6-h stress) and in the precentral medial cortex (1- and 6-h stress). Significantly reduced numbers of GFAP-ir cells were observed only in the medial precentral cortex (1- and 6-h stress); no significant changes were observed in the anterior cingulate cortex. No significant changes of the two glial markers were observed in the hippocampus. Double-labeling experiments with GFAP and pCREB revealed pCREB labeling only in the hippocampus, where the stressed brains (1 and 6 h) displayed significantly reduced numbers of GFAP/pCREB-ir glial cells. The observed downregulation of glia specific marker proteins is in line with our hypothesis that emotional experience can alter glia cell activation in the juvenile limbic system. PMID- 19137573 TI - "Setting paint" analogy for the hydrophobic self-association of tropoelastin into elastin-like hydrogel. AB - Alkaline tropoelastin solutions (pH 11) were optically clear at low temperatures, but a firm gel formed when the temperature was raised to 37 degrees C. Reversion to a clear solution took place if the temperature was lowered to below 20 degrees C within less than 2 h, but not if 37 degrees C was maintained for several hours. The precipitated elastin-like hydrogel thus formed did not visually redissolve at low temperatures. Tropoelastin hydrogel was stable to subsequent washings with alkaline solution at 37 degrees C, but at 4 degrees C some hydrogel redissolved showing that association is at least partly reversible. Washing the hydrogel with neutral 8M urea solution at 4 degrees C dissolved less than 10% of tropoelastin in 24 h. We characterized this phenomenon by combining temperature-controlled light microscopy analysis, 1H NMR spectroscopy (temperature, diffusion, and relaxation time studies), and UV-absorption-based concentration measurements. The self-association of tropoelastin at pH 11 is due to hydrophobic interactions in an emulsion-like system in which the spherules coalesce in a manner like a water based latex paint that forms a durable hydrophobic sheet as water and the organic solvent evaporate. In the present case, the sedimentation and entanglement of the tropoelastin porous sheets means that reverse dissolution is a kinetically slow process. PMID- 19137574 TI - Characterization of the G-quadruplex structure of a catalytic DNA with peroxidase activity. AB - It has been reported that the complexes formed by hemin and some G-quadruplexes can be developed as a new class of DNAzyme with peroxidase activity. This kind of DNAzyme has received a great deal of attention. But to date, the actual G quadruplex structure that can provide hemin with enhanced peroxidase activity is in doubt. Herein, the G-quadruplex structure of CatG4, a 21-nucleotide DNA oligomer which was previously reported to bind hemin and the resulting complex exhibiting enhanced peroxidase activity, was characterized by fluorescence and circular dichroism measurements. The results suggest that the catalytically active form of CatG4 may be a unimolecular parallel quadruplex rather than a unimolecular chair-type antiparallel quadruplex or a multistranded parallel quadruplex. In addition, the fluorescence analysis of labeled oligonucleotides may be developed as a supplementary tool for the study of DNA conformations. PMID- 19137576 TI - Molecular dynamics simulations of xDNA. AB - xDNA is a modified DNA, which contains natural as well as expanded bases. Expanded bases are generated by the addition of a benzene spacer to the natural bases. A set of AMBER force-field parameters were derived for the expanded bases and the structural dynamics of the xDNA decamer (xT5' G xT A xC xG C xA xG T3').(xA5' C T xG C G xT A xC A3') was explored using a 22 ns molecular dynamics simulation in explicit solvent. During the simulation, the duplex retained its Watson-Crick base-pairing and double helical structure, with deviations from the starting B-form geometry towards A-form; the deviations are mainly in the backbone torsion angles and in the helical parameters. The sugar pucker of the residues were distributed among a variety of modes; C2' endo, C1' exo, O4' endo, C4' exo, C2' exo, and C3' endo. The enhanced stacking interactions on account of the modification in the bases could help to retain the duplex nature of the helix with minor deviations from the ideal geometry. In our simulation, the xDNA showed a reduced minor groove width and an enlarged major groove width in comparison with the NMR structure. Both the grooves are larger than that of standard B-DNA, but major groove width is larger than that of A-DNA with almost equal minor groove width. The enlarged groove widths and the possibility of additional hydration in the grooves makes xDNA a potential molecule for various applications. PMID- 19137575 TI - FtsZ condensates: an in vitro electron microscopy study. AB - In vivo cell division protein FtsZ from E. coli forms rings and spirals which have only been observed by low resolution light microscopy. We show that these suprastructures are likely formed by molecular crowding which is a predominant factor in prokaryotic cells and enhances the weak lateral bonds between proto filaments. Although FtsZ assembles into single proto-filaments in dilute aqueous buffer, with crowding agents above a critical concentration, it forms polymorphic supramolecular structures including rings and toroids (with multiple protofilaments) about 200 nm in diameter, similar in appearance to DNA toroids, and helices with pitches of several hundred nm as well as long, linear bundles. Helices resemble those observed in vivo, whereas the rings and toroids may represent a novel energy minimized state of FtsZ, at a later stage of Z-ring constriction. We shed light on the molecular arrangement of FtsZ filaments within these suprastructures using high resolution electron microscopy. PMID- 19137577 TI - High-resolution structures of collagen-like peptides [(Pro-Pro-Gly)4-Xaa-Yaa-Gly (Pro-Pro-Gly)4]: implications for triple-helix hydration and Hyp(X) puckering. AB - Structures of (Pro-Pro-Gly)4-Xaa-Yaa-Gly-(Pro-Pro-Gly)4 (ppg9-XYG) where (Xaa, Yaa)=(Pro, Hyp), (Hyp, Pro) or (Hyp, Hyp) were analyzed at high resolution using synchrotron radiation. Molecular and crystal structures of these peptides are very similar to those of the (Pro-Pro-Gly)9 peptide. The results obtained in this study, together with those obtained from related compounds, indicated the puckering propensity of the Hyp in the X position: (1) Hyp(X) residues involved in the Hyp(X):Pro(Y) stacking pairs prefer the down-puckering conformation, as in ppg9-OPG, and ppg9-OOG; (2) Hyp(X) residues involved in the Hyp(X):Hyp(Y) stacking pairs prefer the up-puckering conformation if there is no specific reason to adopt the down-puckering conformation. Water molecules in these peptide crystals are classified into two groups, the 1st and 2nd hydration waters. Water molecules in the 1st hydration group have direct hydrogen bonds with peptide oxygen atoms, whereas those in the 2nd hydration group do not. Compared with globular proteins, the number of water molecules in the 2nd hydration shell of the ppg9-XYG peptides is very large, likely due to the unique rod-like molecular structure of collagen model peptides. In the collagen helix, the amino acid residues in the X and Y positions must protrude outside of the triple helix, which forces even the hydrophobic side chains, such as Pro, to be exposed to the surrounding water molecules. Therefore, most of the waters in the 2nd hydration shell are covering hydrophobic Pro side chains by forming clathrate structures. PMID- 19137578 TI - Developing an integrated primary care practice: strategies, techniques, and a case illustration. AB - Numerous studies have now demonstrated that integrating behavioral health and medical care can reduce medical costs, improve patient and provider satisfaction, and enhance clinical outcomes. Given this, one might expect that behavioral health programs would be fully integrated into primary care clinics across the country, but in fact integrated primary care programs remain quite rare. One reason for this discrepancy is that implementing such programs has proven to be extraordinarily challenging. Most of the integrated programs that are currently operating successfully are in settings where professionals are all members of the same health care system (e.g., HMOs, the Veterans Administration, Departments of Family Practice, etc.). Many providers, however, are in communities where various services are provided in different locations from different organizations that have very different clinical, administrative, and financial structures. In these situations, the challenges are even greater. The authors describe a set of strategies and techniques providers can use to move their health care system toward a higher level of integration and illustrate how they applied these steps to develop and assess the impact of an integrated primary care program in the state of Rhode Island. PMID- 19137579 TI - The role of the Val57 amino-acid residue in the hinge loop of the human cystatin C. Conformational studies of the beta2-L1-beta3 segments of wild-type human cystatin C and its mutants. AB - Human cystatin C (HCC) is one of the amyloidogenic proteins to be shown to oligomerize via a three-dimensional domain swapping mechanism. This process precedes the formation of a stable dimer and proceeds particularly easily in the case of the L68Q mutant. According to the proposed mechanism, dimerization of the HCC precedes conformational changes within the beta2 and beta3 strands. In this article, we present conformational studies, using circular dichroism and MD methods, of the beta2-L1-beta3 (His43-Thr72) fragment of the HCC involved in HCC dimer formation. We also carried out studies of the beta2-L1-beta3 peptide, in which the Val57 residue was replaced by residues promoting beta-turn structure formation (Asp, Asn, or Pro). The present study established that point mutation could modify the structure of the L1 loop in the beta-hairpin peptide. Our results showed that the L1 loop in the peptide excised from human cystatin C is broader than that in cystatin C. In the HCC protein, broadening of the L1 loop together with the unfavorable L68Q mutation in the hydrophobic pocket could be a force sufficient to cause the partial unfolding and then the opening of HCC or its L68Q mutant structure for further dimerization. We presume further that the Asp57 and Asn57 mutations in the L1 loop of HCC could stabilize the closed form of HCC, whereas the Pro57 mutation could lead to the opening of the HCC structure and then to dimer/oligomer formation. PMID- 19137580 TI - In vitro dexamethasone pretreatment enhances bone formation of human mesenchymal stem cells in vivo. AB - Bone grafting is the current standard of care for treatment of fracture nonunions, while alternative strategies such as bone marrow-derived mesenchymal stem cells are also used. MSCs can be induced towards the osteogenic lineage by in vitro treatment with dexamethasone (dex). This study aimed to determine the optimal duration of dex treatment for osteogenic differentiation of MSCs in vitro and evaluate the effect of this dex pretreatment on bone formation in vivo. To determine the optimal dex treatment, MSCs were cultivated in osteogenic medium for 5 weeks with a varying dex withdrawal schedule, such that MSCs were exposed to dex for either 0, 1, 2, 3, 4, or 5 weeks. During this period, alkaline phosphatase, calcium, and DNA assays, as well as von Kossa staining and morphological observations were performed. One and 2 week dex-treated groups returned to control levels rapidly, whereas 3 and 4 week groups retained higher levels of differentiation markers, with the 4 week group being the highest. Based on these in vitro results, MSCs (with and without dex) and control fibroblasts were seeded into ceramic cubes, cultured for 4 weeks, and implanted into SCID mice, and harvested 6 weeks postimplantation for histologic evaluation. There was no bone formation in fibroblast-seeded controls, little bone formation in control (CS 1), and extensive bone formation (CS 3-4) in dex-treated MSCs. These results indicate that pretreatment of MSCs with dex results in greater bone formation than in untreated controls. PMID- 19137582 TI - A remembrance of Harunori Ishikawa (1935-2008). PMID- 19137583 TI - An in vitro model system for cytoskeletal confinement. AB - The motility, shape, and functionality of the cell depend sensitively on cytoskeletal mechanics which in turn is governed by the properties of filamentous proteins - mainly actin, microtubules, and intermediate filaments. These biopolymers are confined in the dense cytoplasm and therefore experience strong geometric constraints on their equilibrium thermal fluctuations. To obtain a better understanding of the influence of confinement on cytoskeletal filaments we study the thermal fluctuations of individual actin filaments in a microfluidic in vitro system by fluorescence microscopy and determine the persistence length of the filaments by analyzing the radial distribution function. A unique feature of this method is that we obtain the persistence length without detailed knowledge of the complete contour of the filament which makes the technique applicable to a broad range of biological polymers, including those with a persistence length smaller than the optical resolution. PMID- 19137581 TI - Uncoupling protein 2 polymorphisms as risk factors for NTDs. AB - BACKGROUND: Both environmental and genetic factors are involved in the etiology of NTDs. Inadequate folate intake and obesity are important environmental risk factors. Several folate-related genetic variants have been identified as risk factors; however, little is known about how genetic variants relate to the increased risk seen in obese women. Uncoupling Protein 2 (UCP2) is an attractive candidate to screen for NTD risk because of its possible role in obesity as well as energy metabolism, type-2 diabetes, and the regulation of reactive oxygen species. Interestingly, a previous study found that a common UCP2 compound homozygous genotype was associated with a threefold increase in NTD risk. METHODS: We evaluated three polymorphisms, -866G>A, A55V, and the 3'UTR 45 bp insertion/deletion, as risk factors for NTDs in Irish NTD cases (n = 169), their mothers (n = 163), their fathers (n = 167), and normal control subjects (n = 332). RESULTS: Allele and genotype frequencies were not significantly different when comparing NTD mothers, NTD fathers, or affected children to controls. Additionally, the previously reported risk genotype (combined homozygosity of 55VV and 3'UTR 45 bp deletion/deletion) was not present at a higher frequency in any NTD group when compared to controls. CONCLUSIONS: In our Irish study population, UCP2 polymorphisms did not influence NTD risk. Moreover, the prevalence of this allele in other populations was similar to the Irish prevalence but far lower than reported in the previous NTD study, suggesting that this previous finding of an association with NTDs might have been due to an unrepresentative study sample. PMID- 19137584 TI - Microtubule-dependent motility and orientation of the cortical endoplasmic reticulum in elongating characean internodal cells. AB - Motility of the endoplasmic reticulum (ER) is predominantly microtubule- dependent in animal cells but thought to be entirely actomyosin-dependent in plant cells. Using live cell imaging and transmission electron microscopy to examine ER motility and structural organization in giant internodal cells of characean algae, we discovered that at the onset of cell elongation, the cortical ER situated near the plasma membrane formed a tight meshwork of predominantly transverse ER tubules that frequently coaligned with microtubules. Microtubule depolymerization increased mesh size and decreased the dynamics of the cortical ER. In contrast, perturbing the cortical actin array with cytochalasins did not affect the transverse orientation but decreased mesh size and increased ER dynamics. Our data suggest that myosin-dependent ER motility is confined to the ER strands in the streaming endoplasm, while the more sedate cortical ER uses microtubule-based mechanisms for organization and motility during early stages of cell elongation. We show further that the ER has an inherent, NEM-sensitive dynamics which can be altered via interaction with the cytoskeleton and that tubule formation and fusion events are cytoskeleton-independent. PMID- 19137586 TI - Dynamic expression of the Slit-Robo GTPase activating protein genes during development of the murine nervous system. AB - We investigated the expression of the three known Slit-Robo GTPase activating protein (srGAP) genes in the developing murine nervous system using in situ hybridization. The three genes are expressed during embryonic and early postnatal development in the murine nervous system, showing a distinct pattern of expression in the olfactory system, the eye, forebrain and midbrain structures, the cerebellum, the spinal cord, and dorsal root ganglia, which we discuss in relation to Slit-Robo expression patterns and signaling pathways. We also report srGAP2 expression in zones of neuronal differentiation and srGAP3 in ventricular zones of neurogenesis in many different tissues of the central nervous system (CNS). Compared to srGAP2 and srGAP3, the onset of srGAP1 expression is later in most CNS tissues. We propose that these differences in expression point to functional differences between these three genes in the development of neural tissues. PMID- 19137585 TI - Identification and localization of myosin superfamily members in fish retina and retinal pigmented epithelium. AB - Myosins are cytoskeletal motors critical for generating the forces necessary for establishing cell structure and mediating actin-dependent cell motility. In each cell type a multitude of myosins are expressed, each myosin contributing to aspects of morphogenesis, transport, or motility occurring in that cell type. To examine the roles of myosins in individual retinal cell types, we first used polymerase chain reaction (PCR) screening to identify myosins expressed in retina and retinal pigmented epithelium (RPE), followed by immunohistochemistry to examine the cellular and subcellular localizations of seven of these expressed myosins. In the myosin PCR screen of cDNA from striped bass retina and striped bass RPE, we amplified 17 distinct myosins from eight myosin classes from retinal cDNA and 11 distinct myosins from seven myosin classes from RPE cDNA. By using antibodies specific for myosins IIA, IIB, IIIA, IIIB, VI, VIIA, and IXB, we examined the localization patterns of these myosins in retinas and RPE of fish, and in isolated inner/outer segment fragments of green sunfish photoreceptors. Each of the myosins exhibited unique expression patterns in fish retina. Individual cell types expressed multiple myosin family members, some of which colocalized within a particular cell type. Because much is known about the functions and properties of these myosins from studies in other systems, their cellular and subcellular localization patterns in the retina help us understand which roles they might play in the vertebrate retina and RPE. PMID- 19137587 TI - Beneficial effects of sorafenib on splanchnic, intrahepatic, and portocollateral circulations in portal hypertensive and cirrhotic rats. AB - Portal hypertension, the most important complication in patients with cirrhosis of the liver, is a serious and life-threatening disease for which there are few therapeutic options. Because angiogenesis is a pathological hallmark of portal hypertension, the goal of this study was to determine the effects of sorafenib-a potent inhibitor of proangiogenic vascular endothelial growth factor receptor 2 (VEGFR-2), platelet-derived growth factor receptor beta (PDGFR-beta), and Raf kinases-on splanchnic, intrahepatic, systemic, and portosystemic collateral circulations in two different experimental models of portal hypertension: rats with prehepatic portal hypertension induced by partial portal vein ligation and rats with intrahepatic portal hypertension and secondary biliary cirrhosis induced by bile duct ligation. Such a comprehensive approach is necessary for any translational research directed toward defining the efficacy and potential clinical application of new therapeutic agents. Sorafenib administered orally once a day for 2 weeks in experimental models of portal hypertension and cirrhosis effectively inhibited VEGF, PDGF, and Raf signaling pathways, and produced several protective effects by inducing an approximately 80% decrease in splanchnic neovascularization and a marked attenuation of hyperdynamic splanchnic and systemic circulations, as well as a significant 18% decrease in the extent of portosystemic collaterals. In cirrhotic rats, sorafenib treatment also resulted in a 25% reduction in portal pressure, as well as a remarkable improvement in liver damage and intrahepatic fibrosis, inflammation, and angiogenesis. Notably, beneficial effects of sorafenib against tissue damage and inflammation were also observed in splanchnic organs. CONCLUSION: Taking into account the limitations of translating animal study results into humans, we believe that our findings will stimulate consideration of sorafenib as an effective therapeutic agent in patients suffering from advanced portal hypertension. PMID- 19137588 TI - Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide-1-MIMETIBODY: factors that influence productivity and product quality. AB - In an attempt to develop a high producing mammalian cell line expressing CNTO736, a Glucagon like peptide-1-antibody fusion protein (also known as a Glucagon like peptide-1 MIMETIBODY), we have noted that the N-terminal GLP-1 portion of the MIMETIBODY was susceptible to proteolytic degradation during cell culture, which resulted in an inactive product. Therefore, a number of parameters that had an effect on productivity as well as product quality were examined. Results suggest that the choice of the host cell line had a significant effect on the overall product quality. Product expressed in mouse myeloma host cell lines had a lesser degree of proteolytic degradation and variability in O-linked glycosylation as compared to that expressed in CHO host cell lines. The choice of a specific CHOK1SV derived clone also had an effect on the product quality. In general, molecules that exhibited minimal N-terminal clipping had increased level of O linked glycosylation in the linker region, giving credence to the hypothesis that O-linked glycosylation acts to protect against proteolytic degradation. Moreover, products with reduced potential for N-terminal clipping had longer in vivo serum half-life. These findings suggest that early monitoring of product quality should be an essential part of production cell line development and therefore, has been incorporated in our process of cell line development for this class of molecules. PMID- 19137589 TI - A novel system for evaluation of drug mixtures for potential efficacy in treating multidrug resistant cancers. AB - Multidrug resistant (MDR) cancer is difficult to treat. Chemicals that are effective MDR modulators have never exited clinical trials as FDA approved products due to side effects. It has been hypothesized that using a combination of chemotherapeutics with a mixture of MDR modulators (each with different side effects) may lead to useful treatment strategies. Because the experimental space for combination treatments can be large, this space may be impracticable to explore using animal studies. Here we describe an in vitro system based on microfabrication and cell culture that can potentially be used to explore large experimental spaces efficiently. The Microscale Cell Culture Analog (microCCA) concept mimics the body's response using interconnected compartments that represent various tissues or organs. A microCCA is based on the structure of an appropriate physiologically based pharmacokinetic (PBPK) model and emulates the body's dynamic response to exposure to various drugs and chemicals. For this problem we have chosen a microCCA with living cells representing the liver (HepG2/C3A), bone marrow (MEG-01), uterine cancer (MES-SA), and a MDR variant of uterine cancer (MES-SA/DX-5). In proof of concept experiments we found in 24 h "acute" exposures and 72 h treatments that the microCCA system predicts combining the chemotherapeutic, doxorubicin, with cyclosporine and nicardipine, as MDR modulators will have greater efficacy than using doxorubicin by itself or with either modulator alone. This combined strategy is selective in inhibiting MES SA/DX-5 cell proliferation and may prove to be advantageous in vivo by specifically targeting MDR cancer with acceptable side-effects. This cell specific synergy was not observed in traditional 96-well plate assays. By combining the microCCA with a PBPK model, appropriate drug doses and area under the curve exposure for in vivo trials can be extrapolated directly from the results obtained with this device. This device and approach should be useful in screening potential drug/modulator combinations to determine candidate treatments for MDR cancer. Indeed this approach may be useful for in vitro evaluation of human response to a wide range of exposures to mixtures of chemicals or drugs. PMID- 19137591 TI - The financial implications of computed tomographic angiography in DIEP flap surgery: a cost analysis. PMID- 19137593 TI - The effect of fetal tracheal occlusion on lung tissue mechanics and tissue composition. AB - Fetal tracheal occlusion (TO) is currently used to treat severe cases of congenital diaphragmatic hernia (DH). Clinical and experimental studies suggest an improved postnatal outcome, but lung tissue mechanics after TO have not been studied. We determined the effect of TO on mechanical impedance and lung tissue components in a rabbit model for DH. At 23 days of gestation (term = 31 days) either a sham thoracotomy or a diaphragmatic defect was induced. DH fetuses were randomly assigned to undergo 5 days later TO. Fetuses were delivered by term cesarean section to determine lung to body weight ratio (LBWR), dynamic lung mechanics and lung impedance. Airway resistance (R(aw)), elastance (H(L)), tissue damping (G(L)) and hysteresivity (G(L)/H(L)) were calculated from impedance data. Collagen I and III and elastin were quantified histologically. LBWR was significantly increased by TO compared to DH (P < 0.001) and resistance and compliance of the respiratory system (R(rs), C(rs)) were improved as well. TO resulted in a significant decrease of R(aw) comparable to observations in sham fetuses, without effect on lung tissue mechanics H(L), G(L) and hysteresivity. This coincides with a significant decrease of collagen I, III and elastin in comparison to DH fetuses. In this first report on lung tissue mechanics in a rabbit model of DH, TO had a substantial effect on tissue morphology yet this was not mirrored in lung mechanics. We conclude that the effect of TO on lung mechanics without in utero reversal of occlusion, is dominated by airway remodeling. PMID- 19137594 TI - Recurrent severe lower respiratory tract infections in a child with abnormal tracheal morphology. AB - Localized recurrent respiratory infections, leading to severe hypoxia in young children without immunological abnormalities or other risk factors, should raise the suspicion of airway structural abnormalities. In a 24-month-old boy, with recurrent severe post-viral wheezing and a history of RSV-induced bronchiolitis and gastro-esophageal reflux, fiberoptic bronchoscopy demonstrated an abnormal morphology of the distal portion of the trachea, ending in four openings. Computed tomography (CT) scans demonstrated the presence of a right tracheal bronchus and an anomalous upper lobar bronchus, originating at the level of the major carina. PMID- 19137595 TI - Ciliopathy spectrum expanded? Jeune syndrome associated with foregut dysmotility and malrotation. AB - We report the association and surgical management of gastrointestinal dysmotility and malrotation with Jeune asphyxiating thoracic dystrophy (JATD), an autosomal recessive condition that often results in respiratory failure due to a small rib cage. A 4-month-old male with JATD presented with vomiting and aspiration pneumonitis compounding already severe respiratory morbidity. A contrast study revealed esophageal and gastric dysmotility with associated malrotation. This was treated surgically with good results. Some cases of JATD are caused by missense mutations in the gene IFT80, which encodes a protein implicated in the process of intraflagellar transport of primary cilia. We speculate that these abdominal complications might also be part of the extending spectrum of ciliopathy. PMID- 19137596 TI - Journals under threat: a joint response from history of science, technology, and medicine editors. PMID- 19137597 TI - Pharmacokinetics and pharmacodynamics of linezolid in children with cystic fibrosis. AB - Alternative antimicrobial regimens are needed for treatment of methicillin resistant Staphylococcus aureus (MRSA)-associated pulmonary exacerbations in children with cystic fibrosis (CF). There are no published pharmacokinetic (PK) and pharmacodynamic (PD) data for linezolid in children with CF. OBJECTIVES: (1) To determine the PK and PD profile of linezolid among children with CF; (2) to characterize the effect of linezolid on MRSA infection; (3) to determine the effect of age and CF transmembrane regulator (CFTR) gene mutations on drug clearance. HYPOTHESES: Linezolid clearance is enhanced in children with CF requiring a higher dosage regimen. Age and CFTR gene mutations affect drug clearance. METHODS: This was a retrospective cohort study; medical records of children with MRSA-associated pulmonary exacerbations treated with linezolid (10 mg/kg/dose IV every 8h) were reviewed. Linezolid peak and trough concentrations in serum were determined by high performance liquid chromatography, PK profiles determined using standard noncompartmental method, and PD indices were evaluated. RESULTS: 10 children (mean +/- SD, 10.2 +/- 5.5 years) received 14 courses of linezolid at 10 +/- 0.4 mg/kg/dose every 8h for 15.4 +/- 3.2 days. Seven had homozygous DeltaF508 CFTR mutation. Peak and trough linezolid concentrations varied widely (range, 8.4-20.5 and 0.1-11.5 mcg/mL respectively). The PK profile of children <10 years differed significantly from older patients (>or=10 years). The PK indices of children with homozygous DeltaF508 differed marginally from those with heterozygous CFTR mutations, but there were too few subjects to allow separation of age and CFTR mutations effect. No patient achieved the target PD ratio of AUC/MIC >80. MRSA persisted in sputum or throat culture after treatment with linezolid. CONCLUSIONS: Additional PK and PD data are needed to optimize linezolid therapy in children with cystic fibrosis; it is likely that higher doses will be needed. PMID- 19137598 TI - Crystal structure of human osteoclast stimulating factor. PMID- 19137599 TI - Rhesus macaque: a tight homodimeric CD8alphaalpha. PMID- 19137600 TI - Predicting residue-residue contact maps by a two-layer, integrated neural-network method. AB - A neural network method (SPINE-2D) is introduced to provide a sequence-based prediction of residue-residue contact maps. This method is built on the success of SPINE in predicting secondary structure, residue solvent accessibility, and backbone torsion angles via large-scale training with overfit protection and a two-layer neural network. SPINE-2D achieved a 10-fold cross-validated accuracy of 47% (+/-2%) for top L/5 predicted contacts between two residues with sequence separation of six or more and an accuracy of 24 +/- 1% for nonlocal contacts with sequence separation of 24 residues or more. The accuracies of 23% and 26% for nonlocal contact predictions are achieved for two independent datasets of 500 proteins and 82 CASP 7 targets, respectively. A comparison with other methods indicates that SPINE-2D is among the most accurate methods for contact-map prediction. SPINE-2D is available as a webserver at http://sparks.informatics.iupui.edu. PMID- 19137602 TI - Granulocyte-monocyte adsorptive apheresis in pediatric inflammatory bowel disease: results, practical issues, safety, and future perspectives. AB - BACKGROUND: The purpose of the study was to collect data on granulocyte-monocyte adsorptive apheresis (GMA) for the treatment of corticosteroid-dependent (SD) or corticosteroid-resistant (SR) inflammatory bowel disease (IBD) in children from 3 Nordic countries to evaluate its efficacy and safety and to assess practical issues. METHODS: Retrospective data on 37 children treated with GMA were collected. In all, 22 children had ulcerative colitis (UC), 13 Crohn's disease (CD), and 2 had indeterminate colitis (IC). Their mean age was 13.2 years, range 5-17 years, and mean duration of disease was 2.4 years, range 1 month to 6 years. Indication for treatment in the UC group was SD in 11 cases, SR in 6 cases, and other reasons in 5 cases. The corresponding numbers in the CD group were SD in 8 cases, SR in 2 cases, and other reasons in 3 cases. In the IC group, 1 had SD and 1 was refractory to steroids, azathioprine, and infliximab. Efficacy was evaluated by severity indices: the Pediatric Ulcerative Colitis Activity Index (PUCAI) and the Pediatric Crohn's Disease Activity Index (PCDAI) and tapering of corticosteroids. RESULTS: PUCAI and PCDAI decreased significantly in both groups after 3 months (P = 0.0007, P = 0.025). The dosage of corticosteroid was significantly reduced in the UC group by the end of GMA (P = 0.004) and this response continued after 3 months. Relapse was seen in 2 patients with UC and 3 patients with CD after 3 months follow-up. CONCLUSIONS: GMA seems to be an effective and safe treatment in 81% of the SD or SR pediatric IBD patients, especially in those with UC. PMID- 19137603 TI - Conformational preferences of a short Aib/Ala-based water-soluble peptide as a function of temperature. AB - The amino acid Aib predisposes a peptide to be helical with context-dependent preference for either 3(10)- or alpha- or a mixed helical conformation. Short peptides also show an inherent tendency to be unfolded. To characterize helical and unfolded states adopted by water-soluble Aib-containing peptides, the conformational preference of Ac-Ala-Aib-Ala-Lys-Ala-Aib-Lys-Ala-Lys-Ala-Aib-Tyr NH(2) was determined by CD, NMR and MD simulations as a function of temperature. Temperature-dependent CD data indicated the contribution of two major components, each an admixture of helical and extended/polyproline II structures. Both right- and left-handed helical conformations were detected from deconvolution of CD data and (13)C NMR experiments. The presence of a helical backbone, more pronounced at the N-terminal, and a temperature-induced shift in alpha-helix/3(10)-helix equilibrium, more pronounced at the C-terminal, emerged from NMR data. Starting from polyproline II, the N-terminal of the peptide folded into a helical backbone in MD simulations within 5 ns at 60 degrees C. Longer simulations showed a mixed helical backbone to be stable over the entire peptide at 5 degrees C while at 60 degrees C the mixed-helix was either stable at the N-terminus or occurred in short stretches through out the peptide, along with a significant population of polyproline II. Our results point towards conformational heterogeneity of water soluble Aib-based peptide helices and the associated subtleties. The problem of analyzing CD and NMR data of both left- and right-handed helices are discussed, especially the validity of the ellipticity ratio [theta](222)/[theta](207), as a reporter of alpha-/3(10)- population ratio, in right- and left-handed helical mixtures. PMID- 19137604 TI - Cyclosporine treatment of steroid-refractory ulcerative colitis during pregnancy. AB - BACKGROUND: Cyclosporine is considered a safe and effective treatment of severe steroid-refractory ulcerative colitis (UC). However, few data are available concerning its safety profile in pregnant women. We report here the experience of 5 GETAID centers. METHODS: In a retrospective study data on patients with severe UC treated with cyclosporine during pregnancy were extracted from medical records of consecutive patients treated between 2001 and 2007. RESULTS: Eight patients (median age 30.5 years old) were identified. At the time of flare-up the median duration of pregnancy was 11.5 weeks of gestation (range 4-25). Seven patients had pancolitis. All patients had more than 3 commonly used clinical and biological severity criteria. Three patients had severe endoscopic lesions and 5 patients had not. All patients received intravenous corticosteroids for at least 7 days before introduction of cyclosporine. Two patients received azathioprine during treatment with cyclosporine. No severe infections or other complications due to treatment were observed. Treatment was effective in 7/8 patients. One patient received infliximab due to cyclosporine therapy failure with a good outcome. No colectomy was performed during pregnancy. Seven pregnancies were conducted to term, but 1 in utero death occurred due to maternal absence of S protein. Two newborns were premature, including 1 case of hypotrophy. No malformations were observed. CONCLUSIONS: In our experience, treatment with cyclosporine for steroid-refractory UC during pregnancy can be considered safe and effective. PMID- 19137605 TI - Survey of gastroenterologists' awareness and implementation of AGA guidelines on osteoporosis in inflammatory bowel disease patients: are the guidelines being used and what are the barriers to their use? AB - BACKGROUND: The American Gastroenterology Association (AGA) published guidelines to assist clinicians in the evaluation and management of osteoporosis in inflammatory bowel disease (IBD) patients. Two studies suggest that when clinicians utilized the guidelines, the majority of their IBD patients were appropriately screened and treated for metabolic bone disease. The aim was to study whether physicians who say they use the AGA Guidelines are, in fact, following the recommendations, and to assess the barriers preventing the use of the guidelines in the management of osteoporosis in their IBD patients. METHODS: In all, 1000 physicians were selected from the AGA membership list and mailed a survey inquiring into awareness and implementation of the guidelines on osteoporosis in IBD patients. The barriers to implementation of the guidelines were also assessed. The sum of 21 self-reported clinical practices (absence = 0, presence = 1) was used to evaluate adherence to the guidelines. A value of 0 implied no adherence while a score of 21 meant complete adherence. RESULTS: Of 304 responders, 27 fellows, 8 retirees, and 11 incomplete responses were not included in the analysis; thus, 258 respondents were the subject of this analysis. Slightly less than half of the responders used the guidelines in decision-making (126, 49%) or in the management (110, 42%) of their IBD patients. Using the scoring system described above, clinicians self-reporting use of the guidelines had a significantly higher clinical practice score than those who did not use the guidelines (Wilcoxon rank sum test; P < 0.0001). Only 18% (12 of 68) of clinicians whose practice was comprised of 25%) were also more likely (97/187 = 52%) to assess and treat osteoporosis in their IBD patients. Conversely, only 16% (11/68) of physicians who saw